CONCLUSION: The changes of Müller cell shape and the permeability of cell membrane maybe one of factors to trigger the increasing expression of GFAP.

]]> Chuan-Xu Li,Lin-Hui Liang,Lian-Xi Jing,Dao-Man Xiang and Bo-Lin Xie 338true 2O were intravitreously injected. Three days and 14 days later, the animals were sacrificed and the eye balls were cut with cryostat, then TUNEL staining protocol were carried out to detect apoptotic cells and degenerate neurons in the retina. Immunofluorescent double labeling was performed to identify which cells were PEDF-positive cells in the retina.RESULTS:The IOP was obviously increased after the operation(P＜0.05), there was no significant difference in IOP between 3 days and 14 days after the operation. TUNEL-positive cells and FJ-positive cells in retina of chronic intraocular hypertension group were much more than that of normal group. The number of TUNEL-positive cells in retinas of chronic intraocular hypertension+ VEGF group was reducing. CONCLUSION: Intravitreal administration of VEGF has the effect of reducing the apoptosis of the RGCs in rats with chronic intraocular hypertension. ]]> Bing-Jian Lü,Rui-Fu Wang,Xiao-Yun Dong and Xiu-Xiang Ji 337true 570 value. The expression of keratin and HPA-1 in the cells was detected by immunochemistry.RESULTS: HPA-1 was intently expressed in both the cytoplasm and nucleus of human RPE cells, and weak expression was seen in RPE cells co-cultured with PI-88. Either in 48 hours or 72 hours, the A₅₇₀ values of various concentrations of PI-88 culture group were significantly declined in comparison with without PI-88. When PI-88 was administered for 72 hours, the A570 values showed the tendency of reduction to those of 48 hours in >100mg/L groups.CONCLUSION:HPA-1 inhibitor can suppress the proliferation of human RPE cell at the concentration- and time-dependent manner in vitro.]]> Wei-Qiang Tang,Yan Zhang and Xiao-Yan Li 336true 2/M cell cycle arrest and induce apoptosis,especially in the high concentration groups. CONCLUSION: Lithium chloride could restrain growth of HTFs by evoke G₂/M cell cycle arrest and induce apoptosis.]]> Hui-Hui Zou,Ji-Bing Wang,Xu-Dong Huang,Shan-Shan Liu and Hui Man 335true Ya-Lin Ren,Zeng-Zhi Wang and Hao Yan 334true 0.05). Other groups could significantly inhibit HPFs proliferation in a dose- and time-dependent manner, and there were significant differences between each of the two groups (P<0.05). Flow cytometry examination suggested the cell percentage in the G₀/G₁ phase increased while that of the S phase decreased exposed to different concentrations of GBE for 48 hours, the rate of apoptosis of HPFs respectively reached 4.37%, 8.14%, 11.09%, 15.18%, and there were significant differences between the two groups (P<0.05). CONCLUSION: GBE can significantly inhibit HPFs proliferation and induce its apoptosis with dose- and time-dependent manner.]]> Jing Zhu,Chao-Qiong Wu and Dan Chen 333true Wei Chen and Xin Xia 332true 0.05）.VEGF and VEGFR2 dyeing area and intergral optical density in group B were higher than those of group A and group C(P＜0.05).Protein expression in retina in group B was higher than that of group A and group C(P＜0.05). CONCLUSION: It plays a positive role in ameliorating biochemical indicators such as lipid and hemorheology levels in dyslipidemia ApoE-/-mice by using jianpiquyu method.It also plays a regulatory function in the VEGF/VEGFR2 pathway in this age-related macular degeneration (AMD) animal model.]]> Xiao-Hu Liu,Qing-Hua Zeng,Ya Mo,Chun-Yan Tang,Li-Dan Xie and Yi Wang 331true Chun-He Shi and Xiao-Chuan Lan 330true Jing Qu and Dong-Bo Pang 329true 0.05). Compared with sham group ,experimental group had the expression of Nogo-A gradually increased in 12 hours after RIR(P<0.05), peaked in 48 hours(P<0.01),decreased in 72 hours(P<0.05) and reached in normal leavel in 168 hours(P>0.05); the expression of NgR increased in 6 hours after RIR(P<0.05), peaked in 48 hours(P<0.01),decreased in 72 hours(P<0.05) and reached in normal leavel in 168 hours(P>0.05). The expression of Nogo-A and NgR in experimental group showed a positive correlation(P<0.01). CONCLUSION: Nogo-A and NgR expressed at each time point after RIR as a parabola.NgR expressed prior to Nogo-A.They were all peaked at 48 hours after RIR.The expression of Nogo-A and NgR closely related to RIR injury, with a collaborative relationship between the two.]]> Ning Du,Wen-Fang Zhang,Jian-Hua Lu and Dong-Mei Zhang 328true 2+IR,n=32).In the latter two groups,each sub-group was determined by the time after ischemia-reperfusion: 6, 24, 48 and 72 hours. By improving the anterior chamber intraocular pression to set up ischemia-reperfusion model in big rats.The morphology change of retina tissue was detected by light microscope.The changes of Bcl-2, Bax protein were measured by immunohistochemistry in retinal tissue . RESULTS:.In the E₂+IR group, the damage of the retina was significantly reduced than IR group.In IR group,the expression of Bax protein began to add at 6 hours after reperfusion,24 hours reached the peak,48 hours began to reduce,and 72 hours had weak expression. In IR group the expression of Bcl-2 reached the peak at 6 hours,with time going,the expression of Bax began to reduce;but in E2+IR group were increased significantly than IR group in all the time points(P<0.01). CONCLUSION:The retina organization was injured by ischemia-reperfusion injury.Bcl-2 and Bax protein had taken part in the RIRI. Estradiol benzoate could increase the expression of Bcl-2 and cut the expression of Bax protein, and protect the retina tissue.]]> Jian-Hong Lü and Qi Zhang 327true Yu-Qing Chen and Shao-Feng Liu 326true Lin-Ling Li and Min Feng 325true Feng-Li Gao and Xue Qi 324true Dan-Yao Nie and Jia-Qi Chen 323true Zhong-Guo Li,Qing Wang,Hong-Ying Mei,Yi-Qing Luo,Yong-Juan Chen,Yu-Hong Zhang,Ping Yu,Ya-Ge Yun and Qiang Shi 322true Zhi Li,Hao Wang,Suo-Xin Liu,Hui-Ying Liu,Rui-Yi Tan,Jing-Jing Gai and Xue-Hong Ju 321true Dong-Ju Qin,Qing-Hua You and Luo-Sheng Tang 320true th,-8^th and 12^nd week after surgery respectively and processed for light microscope and transmission electron microscope. RESULTS: Throughout the observation period in all groups there was unremarkable findings of the retina,optic nerve,and no corneal edema, intraocular hemorrhage, endophthalmitis were noticed. No statistic difference of intraocular pressure was noticed between control group and two experiment groups or within each group preoperatively or postoperatively. A large clear bubble was formed after Densiron-68,-silicone oil were injected into vitreous cavity. Densiron-68 depicted dispersion beginning between 3^rd and 8^th week. Only one posterior subcapsular lens opacity was observed in Densiron-68 tamponade group. Histological evaluation of the retina exposed to Densiron-68 or silicone oil showed normal retinal structures by light microscopic examination. The mitochondria swollen in the inferior photoreceptor inner segment were observed after 4 weeks in Densiron-68 injection group. The changes mentioned above were more remarkable after 8 weeks. But only a few mitochondria in the photoreceptor inner segment were swollen slightly after 12 weeks. The similar changes were in inner nuclear layer(INL) cell in the superior retina after 12 weeks in silicone oil tamponade group. CONCLUSION: Densiron-68 can be well tolerated by retinas of rabbit, although its propensity to disperse, the availability of Densiron-68 may add to our repertoire in managing selected RD. The mitochondria swollen in the inferior photoreceptor inner segment are observed in Densiron-68 injection groupe. It needs more study to evaluate the damage to anterior chamber and cornea. Densiron-68 should not be used to silicone oil-dependent patient because its propensity to disperse and should be extracted as soon as possible after retina reposition.]]> Xi-Bin Zhou,Xiu-Qin Pang,Lin Li,Jie Yu and Tian-Wei Liang 319true Yi-Jie Li,Xing-Ru Zhang,Min-Hong Xiang,Long Zhang,Qing-Song Li and Zhu-Mei Han 318true 0.05). For the average optical density (gray) measured, PCNA expression in group A and group C were strongly positive, group B was partially positive, positive expression in group D was still less than that in group B. Statistical analysis of group B,D and group A, group B and D were significant difference (P<0.01). Group A and group C were no significant difference (P>0.05). CONCLUSION: In rabbits in vivo, EDTA, poly-L-lysine and EDTA hinge all have functions of inhibiting the proliferation of rabbit LECs, preventing and delaying the formation of PCO. Group D is superior to group B on inhibition of LECs, LECs inhibition of group C is not obvious.]]> Yu-Zhi Bao and Hai-Yan Zhang 317true 0.05). RT-PCR showed that VEGFmRNA was detected in both normal control group and hyperlipidemia group. The expression of VEGFmRNA paralleled to the result of immunohistochemical method. CONCLUSION:The lesion on the choroidal blood vessels and enhanced expression of VEGF caused by hyperlipidemia might lead to CNV.]]> Hong Lin,Guo-Xing Xu,Jian Guo,Mao-Song Xie and Xue-Dong Zheng 316true Xu-Tao Chen and Zhan-Yu Zhou 315true Bo Zhang,Dong-Bo Pang and Qing-Yun Meng 314true Hui Zhong,Wang Fang,Ling-Yan Chen,Li Zhang,Li Wang and Shi-Yi Xiao 313true rdth day, reached the peak within the 7^th-10th day, and decreased after the 14^th day. The CNV in cornea after alkaline burn was found on the 3^rd day, increased on the 5^th day, reached the peak of area within the 7^th-10^th day, and decreased after the ^14th day. The expression of ILK and VEGF showed positive correlation at different time points (r=0.926，P<0.01). And they were positively related to CNV (ILK and area of CNV, r=0.900, P<0.01; VEGF and area of CNV, r=0.878, P<0.01). CONCLUSION: ILK and VEGF were expressed significantly in cornea of rats after alkaline burn. And they involved in the regulation of CNV.]]> Peng-Fei Dai,Feng Wang,Yu-Ping Zheng,Yan-Long Quan and Jing-Jing Zhang 312true Cheng-Ye Che,Ying-Ying Mu,Qiang Xu,Na Li,Wen-Yan Jia,Cui Li,Qiu-Qiu Zhang,Qing Wang and Gui-Qiu Zhao 311true 2O₂） and experimental groups(（H₂O₂+TP）were set respectively.After 6, 12, 24 and 48 hours, the cloudy conditions of lens were observed in every group, and the superoxide dismutase(SOD), glutathione peroxidase(GSH-Px) activity and malondaldehyde(MDA) content were detected of each lens. RESULTS: Untreated group-s lens all maintained transparent. Along with the extension of action time, cloudy degree of the lens gradually increased obviously in oxidative damage group.The lens cloudness in tea polyphenols group was less than that in oxidative damage groups. The differences among groups in lens opacity relative grey value were statistically significant （P<0.05）. MDA content of lens tissue in oxidative damage group was significantly higher than that in control group （P<0.05）,while GSH- Px and SOD decreased significantly （P<0.05）.TP group being compared with oxidative damage group, its MDA in lens reduced （P<0.05）, but still higher than that in control group, and GSH-Px, ATP levels increased obviously(P<0.05),all the differences were statistically significant. CONCLUSION: TP can improve the lens oxidation resistance to oxidative damage, reduce lipid peroxide level, thus delay the origin and development of cataract, and provide new theory basis for the clinical diagnosis and treatment of cataracts.]]> Jia Li and Dan Liu 310true Han Bing,Jie Tang,Xiao-Ling An and Hong-Dan He 309true Er-Xia Wei,Xiao-Yun Ke and Rui-Hong Ju 308true Mei-Lin Fu,Qing-Hua Peng,Ji Chen and Yan-Fei Xing 307true 2) on the expression of TGF-β² mRNA in human Tenon-s capsule fibroblasts. METHODS: Human Tenon- s fibroblasts were transferred by TGF-β² siRNAs vector after separated, cultured and passaged three in vitro. RT-PCR was performed to evaluate the levels of TGF-β² mRNA at 24, 48 and 72 hours after transferred in transferred cells, and untransferred cells were as the controls. RESULTS: Separated human Tenon-s fibroblasts attached in 4 hours and confluent monolayer cells formed in 36 hours. The adherent cells displayed fibroblast-like or spindle shape by microscope observation. The expression of TGF-β2 mRNA of transferred group was reduced significantly in comparison with the control group，expression inhibition rates were 17.40%, 52.80% and 79.20% respectively in 24, 48 and 72 hours after transferred,and this interference function was strengthened with the time extending. CONCLUSION:The vector of siRNA specific for TGF-β² is of potent interference ability for TGF-β² mRNA expression in human Tenon-s capsule fibroblasts.]]> Fang Qiao,Fang-Ting Zhang,Pei Fu and Ming-Hua Li 306true 2S) on cell apoptosis and expression of Bcl-2/Bax in rat retinal ischemia-reperfusioninjury(RIRI). METHODS:A total of 54 SD rats were randomly divided into normal control group,ischemia-reperfusion model group and sodium hydrosulfide(NaHS) treatment group,the last two groups were subdivided into group 6, 24, 48, 72 hours after reperfusion. The models of RIRI were made by elevating intraocular pressure.Apoptosis was assessed by the terminal deoxynucleoitidyl transferase mediated dUTP nick end labeling method,and the expression of Bcl-2/Bax was studied by immunohistochemistry. RESULTS:There was a significant number of TUNEL positive cells 6 hours after transient ischemia peaked at the 24 hours, followed by a decrease at the 48 hours. Compared with RIRI group, Bcl-2 protein expression was increased, Bax protein level were decreased in NaHS group (all P<0.05). CONCLUSION: H₂S can enhance the expression of Bcl-2,decrease the expression of Bax, increase the Bcl-2 /Bax ratio and decrease the apoptosis of ganglion cells. H₂S shows a significant protection for retinal ganglion cells with RIRI.]]> Peng-Peng Liu and Qi Zhang 305true METHODS:RGCs were cultured in DMEM(control group)or DMEM containing rhEPO(rhEPO group)for 72 hours. Cell morphology and axonal growth were observed under phase-contrast microscope. The length of the longest processes of RGCS were measured and compared between two groups. The GAP-43 expression was detected by Western blot and the gray value scales of GAP-43 were measured by imaging analysis system. All results were compared using t test.

RESULTS:The RGCs were observed to extend processes under microscope after culturing for 72 hours. The cells in rhEPO group were bigger and the processes were longer compared to the control group(P<0.05). The expression of GAP-43 in vitro was detected by Western blot. The expression of GAP-43 in rhEPO group was higher than that in control group(P<0.05).

CONCLUSION: rhEPO can promote the axonal growth of cultured RGCs in vitro. rhEPO can also increase the expression of GAP-43 in cultured RGCs, which may result in the promotion of rhEPO on the axonal growth of RGCs.]]> Hui Wang and Zhe-Li Liu 304true METHODS: Fresh eyeballs HRCECs extracted just after cornea transplanting operation were cultivated in vitro. The third or fourth generation healthy cells for this experiment were obtained, and divided into three groups: low glucose, high glucose, high glucose plus different density of Rhizoma Zedoariae group(60, 80, 100, and 120μg/mL). MTT was employed to detect the multiplication of HRCECs, and the HRCECs VEGF through immune cells was observed.

RESULTS: MTT comparison: there was no obvious difference between high glucose and low glucose groups(P>0.05), while there was significant difference between high glucose+ and high glucose groups(P<0.05), and different density of glucose showed big difference. Immune cells: high glucose group had obvious affection compared with low glucose group, and there was significant difference between high glucose+ group and high glucose group in their affection on VEGF(P<0.05), as well as in different density among high glucose group.

CONCLUSION: Rhizoma Zedoariae can restrain the multiplication of HRCECs and VEGF in high glucose environment.]]> Hong Tang,Hui-Can Peng,Qi-Lin Cheng and Wen-Wen Xiang 303true METHODS:The method of subretinal injection of Matrigel on grey rabbits was used to build up the model of experimental CNV. Fundus examination, FFA, choroidal flat mount and pathological changes were measured to choose the best dose and injecting technique of Matrigel.

RESULTS: In the FFA examination, the fluorescence leakage was happened in Matrigel 40μL and 20μL groups at 7 day, which was enhanced at 14 day and 28 day; the fluorescence leakage was happened in Matrigel 10μL group at 14 day and 28 day,which was never found in Matrigel 5μL group. In the fundus examination, most of retinas were attached back at 7 day; a few of those were detached. A few of retina had proliferative detachment in Matrigel 40μL group. Cataract and vitreous muddy were found in Matrigel 40μL group. In the tissue pathological examination, small vessels and/or immature fibrous tissues were found in Matrigel 40μL and 20μL groups. The retinal injure was most severely in Matrigel 40μL group.

CONCLUSION: The method of subretinal injection of Matrigel could create CNV successfully, 20μL is the best dose.]]> Zhi-Qiang Wang,Wei Jiang,Li-Na Liang,Shang-Kun Zhou and Ke Song 302true METHODS:Different concentration of free iron, different concentration and variety classes of synthesis iron and biological iron chelating agent were added into pseudomonas aeruginosa biofilm, and then the clump count on the biofilm was detected.

RESULTS:Low concentration of free iron increased the adhesive capacity of the bacterial, with 10μmol/L of iron had the most obvious effect. 0.1μmol/L of EDTA and lactoferrin significantly inhibited the bacterial count of pseudomonas aeruginosa biofilm.

CONCLUSION: Low concentration of free iron can promote the bacterial count of pseudomonas aeruginosa biofilm on the contact lens, the lack iron circumstance which formed by finite concentration lactoferrin and EDTA can prevent the formation of the biofilm.]]> Yan-Ying Wang,Su-Hua Liao and Shu-Nan Zhang 301true METHODS: Forty colored rabbits were selected and divided into 5 groups randomly, including blank group, model group, vitamin E group, zhujingwanjiajianfang group and the holotrichia extractive group. There were 8 rabbits in each group(16 eyes). CNV model was established through the method of argon laser photo-coagulation. Then optic fundus colored pictures were taken after 24 hours, 7, 14, 21 and 28 days, fluorescence fundus angiography(FFA)was performed after 7, 14, 21 and 28 days and optical coherence tomography(OCT)was performed after 14 and 28 days. Then the rabbits were divided into 2 parts each group randomly, and executed by aeroembolism after 14 and 28 days. Their eyes' posterior segment tissues were selected to carry on organized microtome section, hematoxylin and eosin staining and Ang1, PEDF immunohistochemistry staining to analyze depressant effect of holotrichia extractive to CNV.

RESULTS: Determination of Ang1 content expression showed, blank group's content was the least in all groups(P<0.05), the content of holotrichia extractive group and zhujingwanjiajianfang group were less than others in experimental groups(P<0.05), holotrichia extractive group's content was less than zhujingwanjiajianfang group's content, but there was no significant difference, holotrichia extractive group's content between the 14 days and the 28 days has significant difference(P<0.05). Determination of PEDF content expression showed, blank group's content was the highest in all groups(P<0.05), the content of holotrichia extractive group and zhujingwanjiajianfang group were higher than others in experimental groups(P<0.05), holotrichia extractive group's content was higher than zhujingwanjiajianfang group's content, but there was no significant difference, holotrichia extractive group's content between the 14 days and the 28 days had significant difference(P<0.05).

CONCLUSION: The holotrichia extractive has active influence for the high experiment of Ang1 in CNV of the experimental colored rabbits, it can intervene reduction of PEDF and protects retinal tissue, and thus it has depressant effect to CNV.]]> Xiao-Xing Qiu,Qing-Hua Peng,Mei Chen,Jun Peng,Han-Yu Tan and Wen-Juan Li 300true METHODS: Totally 125 Balb/c mice were scratched the word of "#" in cornea using the back of a razor blade cutting edge after anesthesia under the microscope. Among the total of 100 mice were inoculated with herpes simples virus type-Ⅰ. The rest of 25 mice were not inoculated with HSV-Ⅰas normal control group. Using 10g/L sodium fluorescein stained and slit lamp microscope observed the occurrence of corneal disease after operation everyday. Corneal surface tears were taken to carry on the detecting of human embryonic kidney epithelial cells(HEK293T)to define whether the viral replication. The latently infected mice model were exposed to ultraviolet-B light(UV -B)to induce the recurrence of HSK.

RESULTS: The eyes of 100 mice model which inoculated with HSV-Ⅰwere all appeared acute epithelial keratitis within 3 days after inoculation. The corneal inflammation disappeared after acyclovir eye drops treating for 1 week, but the virus detection of polymerase chain reaction(PCR)in cornea and trigeminal ganglion remained positive. The latently infected mice model were exposed to UV-B to induce the recurrence of HSK within 1 week, recurrent HSK were in the form of stromal HSV keratitis principally.

CONCLUSION: Balb/c mice were inoculated with HSV-I virus by corneal scarification, through UV-B rays establishing infected, latent, recurrent herpes simplex keratitis mice model, and the operation is relatively simple, convenient and easy.]]> Ai-Qin Nie,Lei Xi,Xing-Hua Xi,Lu-Yun Lei and Fang Cheng 299true METHODS: Rabbit cornea alkali-burns model was made, then 50 rabbits were randomly divided into experimental group(n=25)and control group(n= 25). At the same time the surgery of conjunctival flap covering was given to rabbits of the experimental group. The condition developing of alkali-burned cornea was observed by slit-lamp biomicroscopy, photography in two groups, and the expression of MMP-9 and TIMP-1 was identified by immunohistochemisty in different period.

RESULTS: Expression of MMP-9 increased on the 3^rd day, shown a peak on the 14^th day, and then decreased gradually. TIMP-1 expressed in the early phrase, and later, the level decreased on 7^th day. TIMP-1 level reached the lowest level on 14^th day, then increased and shown a peak on the 21^st day. MMP-9 level of the experimental group was significantly lower than that of the control group(the 3^rd, 14^th, 21^st, and 28^th day), while TIMP-1 level(the 3^rd, 14^th and 21^st day)was higher(P<0.05).

CONCLUSION: During the wound healing process, alkali-burned cornea has close relation with the expression of MMP-9 and TIMP-1. The treatment of conjunctival flap covering for the severe alkali-burned cornea was found to have good effect.]]> Dong-Yu Song,Ming-Hong Gao,Xu Xu,Jing Yu,Hai Yu,Ying-Xin Chen and Xiao-Guang Zhang 298true METHODS: Effects of estrogen on the expression of PEDF and VEGF in the cultured rat retinal Müller cells were investigated with the methods of RT-PCR and Western blotting analysis.

RESULTS: The levels of PEDF mRNA and protein decreased and VEGF mRNA and protein increased when the Müller cells were under hypoxic condition for 24 hours. Estrogen had protective effect on Müller cells.

CONCLUSION: Estrogen can regulate the expression of PEDF which may play an important role on retinal neovascularization.]]> Hua Mu,Xiao-Mei Zhang,Zhuo-Lei Feng and Ye-Qing Wang 297true METHODS: Form deprivation amblyopia rats models were established, the morphological changes of visual cortex area 17 in 10 normal rats and 10 form deprivation amblyopia rats were stained with HE and observed HE staining, processed by immunohistochemical staining and image analysis system to demonstrate and analyze the localization of caspase-3 in area 17.

RESULTS: At various levels of the form deprivation amblyopia rats and normal rats, there were lots of caspase-3 immunoreactive neurons in visual cortex area 17, especially Ⅱ-Ⅳlayer. The number and grey level of caspase-3 immunopositivity increased greatly than normal rats(P<0.05).

CONCLUSION: Caspase-3 in visual cortex area 17 of form deprivation amblyopia rats has increased expression, which may be involved in the development of amblyopia.]]> Ya-Wei Zeng,Xiang-Zhen He and Zhi Wang 296true METHODS:It was an controlled experimental study, using 200μmol/L CoCl₂ treated RPE cells to establish chemical hypoxia model. The expression of DDR2 in cells was examined after hypoxia 0, 2, 6, 12, 24 hours by immunofluorescence, reverse transcription polymerase chain reaction(RT-PCR)and Western blotting. In hypoxia state with collagenⅠ's(10μg/mL in 50 degrees incubated box )stimulation, the expression of DDR2 and HIF-1α in cells were examined after hypoxia 0, 2, 6, 12, 24 hours by RT-PCR and Western blotting, using enzyme linked immunosorbent assay(ELISA)to observe the expression of VEGF in cell culture supernatant.

RESULTS:After prolonged hypoxia, the expression of DDR2 mRNA and protein were reduced. CollagenⅠcould multiply the expression of DDR2 mRNA and protein in hypoxia as time goes on, it could active DDR2. The expression of HIF-1α mRNA, protein and VEGF protein dropped off in hypoxia state with collagenⅠ's stimulation compared with hypoxia.

CONCLUSION: The expression of DDR2 in RPE cells reduced with time under hypoxia state. CollagenⅠcan activate DDR2 as time goes on. In hypoxia state with collagenⅠ's stimulation can surpress hypoxia induced up- regulation expression of HIF-1α and VEGF in RPE cells.]]> Yu Zhang,Jie Zhu,Zhao-Jie Chu and Yu-Sheng Wang 295true METHODS:Randomly divided the lacrimal gland epithelial cells in good condition into the flavonoid treatment group without androgen cultural(hereinafter referred to as Buddleia flavonoids group), the control group with androgen cultural(hereinafter referred to as androgen group), and the blank group without androgen cultural(hereinafter referred to as the blank group). Total flavonoids with Buddleia plasma, testosterone propionate and blank plasmain were added to begin the interposal. Then, cell models of dry eye syndrome were established. Trizol reagent was used to extract the whole cell RNA. RT-PCR method was used to detect the expression of the Bax mRNA and Bcl-2 mRNA in lacrimal gland epithelial cells, and the results were contrasted.

RESULTS:The Buddleja group and the androgen group inhibited the expression of Bax mRNA in lacrimal gland epithelial cells, compared with the blank group the difference was significant(P <0.01), the expression of Bax mRNA in the Buddleia group is lower than that in the androgen group, the difference was significant(P <0.01), however, the expression of Bcl-2 mRNA in the Buddleia group and the androgen group were higher than that in the blank group, the difference was significant(P <0.01). The expression of Bcl-2 mRNA in Buddleia group is higher than the androgen group, the difference was significant(P <0.01).

CONCLUSION:Buddleja role of flavonoids in the drug- containing plasma androgen levels decrease in cell models of dry eye syndrome can inhibit the expression of Bax mRNA, decrease the amount of the expression of Bax mRNA, and promote the expression of Bcl-2 mRNA, increase the expression of Bcl-2 mRNA. It is possible that the Buddleja flavonoids drug-containing plasma achieve the inhibition of apoptosis by inhibiting the expression of the Bax mRNA and promoting that of Bcl-2 mRNA in lacrimal gland epithelial cells.]]> Fen Wang,Qing-Hua Peng,Hai-Zhong Li,Fang Wang,Xiao-Lei Yao and Wen-Juan Li 294true 35H₂₇O₅)₈ mediated PDT on retinal pigment epithelial cell(RPE).

METHODS: hRPE cells were divided into blank control group, laser group, photosensitizer SiPc(C₃₅H₂₇O₅)₈ group, PDT 1J group, PDT 2J group. CCK-8, fluorescent microscopes blue excitation were applied in the experimental groups, and active oxygen level of RPE cell mitochondria and RPE cell apoptosis rate were detected by DCFH-DA.

RESULTS: Different concentration photosensitizer \〖SiPc(C₃₅H₂₇O₅)₈\〗 mediated PDT on RPE had different effects, in which 5 μg/L of concentration had the most serious damage.

CONCLUSION:New photosensitizer SiPc(C₃₅H₂₇O₅)₈ mediated PDT on RPE has certain damage effect, its possible mechanism is that the increasing RPE cell mitochondria reactive oxygen species level caused by photosensitizer \〖SiPc(C₃₅H₂₇O₅)₈\〗 mediated PDT results in cell apoptosis.]]> Guo-Xing Xu,Xiao-Fang Zhou,Li-Min Chen and Yi-Ru Peng 293true in vitro.

METHODS: The expression of TK gene in lens epithelial cells mediated by rAAV2/HSV-tk was detected by RT-PCR. MTT assay was used to observe the changed bystander effects of HSV-tk/GCV system in N/N1003A cells after ATRA therapy. The effects of ATRA therapy on the apoptotic rates of HSV-tk/GCV system in N/N1003A cells were investigated by flow cytometry.

RESULTS: HSV-tk gene mediated by rAAV2 was transfected successfully into N/N1003A cells and expressed stably. The bystander effect of HSV-tk/GCV system on N/N1003A cells in ATRA treatment group was significantly improved when compared with control group(P<0.01), and the apoptotic rates of the study group were notably increased(P<0.01).

CONCLUSION: ATRA can significantly enhance the bystander effect of HSV-tk/GCV suicide gene system mediated by rAAV2 in rabbit lens epithelial cells in vitro.]]> Zhi-Xiang Ding,Yan-Yi Peng,Wen-Bin Zhang,Mei-Yuan Qiu and Yu-Ming Zhang 292true METHODS: EAU was induced in C57BL/6 mice by subcutaneous immunization of CFA+PTX+IRBP in hind limb and empennage. Peripheral blood flowcytometry was performed at 3, 7, 14, 21 and 28 days after immunization, respectively.

RESULTS: After immunization with specific antigen IRBP, clinical assessment showed EAU occurred at 14 days and became severe at 21 days, but gradually relieved at 28 days. Accompanying with intraocular inflammation of EAU, both CD4⁺CD25⁺ T regular cells and CD4⁺CD3⁺ helper T cells increased. Especially,CD4⁺CD25⁺ T regular cells increased more obviously at 21 days. The number of CD4⁺CD25⁺Foxp3⁺ T regular cells had a great raising and gradually reduction after 28 days. The ratio of CD4⁺CD25⁺Foxp3⁺/CD4⁺CD25^-Foxp3⁺ had a piecemeal increase after 3 days and reached the peak at 21 days then reduced at 28 days.

CONCLUSION: The groups of CD4⁺CD25^+/- Foxp3⁺ T regular cells have a close relationship with the development and turnover of EAU. The groups of CD4⁺CD25^+/-Foxp3⁺ T regular cells provide a new thread for clarity remission mechanism of EAU, prevention and treatment of EAU.]]> Li-Wei Qin,Xiu-Jun Peng,Jian-Wei Guo,Gui-Qin Wang,Yuan Gao and Li-Qun Cao 291true 2 of guinea pig retinal pigment epithelial(RPE)cells.

METHODS: Ten two weeks old healthy guinea pigs, RPE cells were cultured conventionally and divided into four groups: focused light group, defocused light group, parallel light group and the control group. The first three groups were exposed to the focused light, defocused light(passed though lens from parallel light)and the parallel light, the control group was untreated. The level of the expression of TGF-β₂ protein and TGF-β₂mRNA was measured by immunocytochemistry and real-time fluorescence quantitative chain reaction 24 hours after light exposure and the pattern-effect relations were explored. The data were analyzed by means of analysis of variance(ANOVA).

RESULTS: The level of both TGF-β₂ protein and TGF-β₂ mRNA were significantly increased in the focused light exposed group compared with the other groups, followed with the defocused group(P<0.05).

CONCLUSION: It was suggested that different forms of light can affect the expression of TGF-β₂ in guinea pig RPE cells, and was most obvious in the focused light exposed group.]]> Jiang-Yuan Qin,Chao-Ying Wang,Ying-Qing Liu,Tao Jin and Chun-Mei Tong 290true METHODS: Totally 42 male Wistar rats were divided into seven groups, six rats in each group and one of the groups as control group. The experimental groups were made into cornea model by needle puncture at their corneas of right eyes. The corneas were surgically removed at six different time points(6h, 1d, 3d, 5d, 7d, 14d)after operation. The localization and expression of BMP-7 was determined by immunohistochemical methods and computer image analysis.

RESULTS: The expression of BMP-7 of experimental groups gradually rose on the 1^st day and 3^rd day in the injured corneal epithelium after operation and significant difference(P< 0.05). Then it decreased gradually on the 5^th day. And it reached the normal level on the 14^th day.

CONCLUSION: BMP-7 is expressed in cornea of rats and may play a negative role on corneal wound healing after injury.]]> 2013/8/26 15:29:05 Sheng-Ju Chen,Wan-Na Ren,Cai-Li Hu,Ying Pu,Li Li and Yu Du 289true METHODS: Healthy male Wistar rats were selected and randomly divided into diabetic group and normal group(M0). Type 1 diabetes rat model was induced by being injected of large-dose streptozotocinum(STZ)into abdominal cavity. The retina and choroid of rats was obtained to detect the VEGF and PEDF expressions by immunohistochemistry at 1^st(M1), 2^nd(M2), 3^rd(M3), 5^th(M5)month after diabetes induction.

RESULTS:VEGF expression in retina and choroid of rats had no obvious difference between M1and M0. M2 VEGF expression in choroid was positive(33.3%), while in retina was not significantly different with M0. M3 VEGF expression in choroid was positive(55.6%), and in inner nuclear layer and ganglion cells layer of retina was positive(33.3%). M5 VEGF expression in choroid was and the whole retina positive(88.9%). PEDF expression in retina of rats had no obvious difference between normal group and M1, M2 group, and there was no PEDF expression detected in choroid of all the groups. In M3 and M5 group, PEDF expression in retina was lower than M0, the expression gradually weakened with the extension of the course(P<0.05), and there was also no PEDF expression detected in choroid of all the groups. The ratio of VEGF/PEDF gradually increased with the extended duration of diabetes, and there was significant difference compared with M0(P<0.01)

CONCLUSION: The expression of VEGF and PEDF in retina of rats was related with the diabetes course, VEGF expression in retina and choroid was increased, while PEDF expression was gradually decreased in the course. There was no PEDF expression in choroid, it might because the experimental model was not in proliferative stage.]]> 2013/8/26 15:29:05 Lin Li,Xi-Yuan Zhou and Ji-Han Luo 288true METHODS:A total of 60 female rats were examined for the expression of MMP-1 and TIMP-1 protein in lacrimal gland through SⅠt measurement together with Western- blot and RT-PCR methods.

RESULTS:There was significant reduction in SⅠt and increase in the protein expression of MMP1, TIMP1 and mRNA after ovariectomy conducted on rats(P<0.05). After estrogen treatment, there was significant reduction in SⅠt and increase in the expression of MMP1, TIMP1 and mRNA as compared with before treatment(P<0.05).

CONCLUSION: Estrogen must be applied cautiously to treat xerophthalmia following menopause because estrogen can up regulate the expressions of MMP-1 and TIMP-1 and can aggravate the decrease of lacrimal secretion.]]> 2013/8/26 15:29:05 Tao Wang,Tao Liu,Hong-Ya Wu,Yang Liu and Jian-Kang Chen 287true METHODS: Fifteen of the forty-five 3-week old female guinea pigs were randomly selected as the blank control group, the rest 30 guinea pigs were using facemask to induce high myopia in 4 weeks, then randomly assigned to model group and treatment group, then induced CNV by different wavelength laser. The treatment group gavaged with modified Zhu Jing prescriptions while the control group was gavaged with distill water. 21 days after treatment, histopathologic choroidal flat mounts, HE stain,immunofluorescence stain and immuno- histochemistry for VEGF in the experimental eyes and control eyes were performed.

RESULTS:All the 30 guinea pigs were successfully induced high myopia and the axial length of the right eyes were significantly greater than the non-facemask eyes and the eyes of control group. Immunofluorescence stain and immunohistochemistry for VEGF showed that the expression of VEGF of the treatment group was less than the model group. The average optical density(AOD)of the form deprivation eye in the treatment group(0.0589±0.0146)was higher than that in the model group(0.0972±0.0507)(P<0.01).

CONCLUSION: Modified Zhu Jing prescriptions has a positive effect to the reduction of the VEGF expression in the CNV with pathological myopia and thus it can inhibit the growth of CNV.]]> 2013/7/29 14:34:24 Nan-Nan Tian,Ze-Feng Kang,Qing Zhang,Wei Jiang,Ling Li and Yong-Gui Wang 286true METHODS: Forty eyes from 40 Sprague-Dawley male rats were used. After alkali injuries at central corneal using 1mol/L NaOH,the control group(Group A)received topical PBS four times a day. The rest groups(Group B, C, D)received topical AM suspension with different concentration(240, 400, 560μg/mL). Using slit lamp biomicroscopy, CNV were evaluated and scored. Tissue ultrastructure changes of cornea were analyzed by corneal confocal microscope.

RESULTS:The area of CNV were statistically different at each time point between group C and group A, B(all P<0.05). CNV were reduced as AM concentrations increased, while there were no significant differences between group C and group D. Inflammatory cells in corneal were reduced in group AM-treated groups.

CONCLUSION: AM suspension leads to significant reduce in CNV with increasing AM concentrations. However, when the concentration reached 400μg/mL, the effects on restraining CNV cannot be enhanced by concentration. This effect may be elicited in part through the inhibiting of inflammatory cells.]]> 2013/7/29 14:34:24 Fen Ye,Feng Jiang,Yu-Hua Shi and Zhen-Ping Huang 285true METHODS: Cells of 8-DC, GFP-DC and IL-10-GFP-DC were prepared. Totally 55 SD rats were randomly selected 6(6 eyes), as a negative control group, the rest were randomly divided into 4 groups: positive control group, injected of 1mL PBS by receptor tail vein 3 days before surgery, and 8-DC group, GFP-DC group, IL-10-GFP-DC group, respectively, injected of 1mL cell suspension(8-DC, GFP-DC and IL-10-GFP-DC)have been prepared by receptor tail vein 3 days before surgery. Wistar rats as donor, keratoplasty was performed. The rats corneal graft status was examined after operation, and ELISA method was used to detect the expression of IL-4, IFN- γ in aqueous humor at the fourteenth day after operation.

RESULTS: The survival time of corneal graft in group IL-10-GFP-DC was significantly prolonged. Expression of IL-4 in group IL-10-GFP-DC cytokines was higher than that in other groups, while the expression of IFN- γ was lower than other groups, the difference was statistically significant.

CONCLUSION: Exogenous IL-10 gene transfection to immature dendritic cells could inhibit corneal allograft rejection, and induce the formation of immune tolerance. The disturbance over the expression of cytokines IL-4, IFN- γ in aqueous humor is one of the mechanisms for the induction of immune tolerance in corneal transplantation.]]> 2013/7/29 14:34:24 Li-Ying Zhao,Shu Zhang,Li-Hua Tian and Bing Li 284true METHODS: The rats were randomly divided into two groups, 20 mice received injection of botulium toxin B(0.1mL), and 10 mice were injected physiological saline(0.1mL). Schirmer I test, corneal fluorescein staining and the levels of MIF, IL-6,TNF-α were performed before and 3, 7, 28, 42 days after injection. The levels of MIF, IL-6, TNF-α were detected by enzyme-linked immunosorbent assay(ELISA).

RESULTS: The tear production was significantly decreased at 4 points and the corneal fluorescein staining increased at 5 points in BTX-B-injected mice compared with control mice. In the BTX-B-injected mice, the level of IL-1β increased significantly at the 3 days and 1, 4, 6 week, and the level of MIF in lacrimal gland increased significantly since the 4^th week compared with control mice. The level of TNF-α has no difference between the two groups.

CONCLUSION: Injection botulium toxin B can successfully established the mice model of dry eye. This model has the characteristic changes of the expression of inflammatory factors, which is an ideal animal model for dry eye experiment.]]> 2013/7/29 14:34:24 Shuai Zhao,Qian-Yan Kang and Wei Gao 283true METHODS: Thirty-two eyes of sixteen New-Zealand rabbits(2.2-2.4kg)were divided into two groups. The left eyes of rabbits underwent standard glaucoma trabecular resection were treatment group, and the normal right eyes served as controls. Transparency of lenses was monitored by a slit-lamp biomicroscopy before and after glaucoma trabecular resection. The morphology of lens cells was observed under the light microscope.The activities of Na⁺-K⁺-ATPase,catalase(CAT), glutathion peroxidase(GSH-px), glutathione reductase(GR), superoxide dismutase(SOD)and content of malondialdehyde(MDA)in lenses were detected six months after trabecular resection.

RESULTS: Lenses were clear in both treatment group and normal control group during the six months after operation. The morphology and structure of lens cells were normal under the light microscope in both operation group and normal group. The activity of lens cells antioxidant enzyme activity were significantly decreased in operation group compared with control group, Na⁺-K⁺-ATPase declined by 20.97%, CAT declined by 16.36%, SOD declined by 4.46%, GR declined by 4.85%, GSH-px declined by 10.02%, and MDA increased by 16.31%.

CONCLUSION: Glaucoma trabecular resection can induce the change of Na⁺-K⁺-ATPase, CAT, GSH-px, GR, SOD and MDA in lens of rabbit. Glaucoma filtration surgery for the occurrence of cataract development mechanism has important guiding significance.]]> 2013/7/29 14:34:24 Jian-Ping Wang,Yong Ma,Yu-Shun Xue,Xuan-Yi Che,De-Xiu Zhang,Tao Zhu,Jing Li,Rui Shi and Gui-E Zhao 282true METHODS: Children were divided into 3 groups according to the position of eye and the type of strabismus as follow: concomitant esotropia group(67 cases), concomitant exotropia group(129 cases)and normal eye position group(23 cases). For each group of the children, the extraocular muscle were cut out and frozen for detecting the expression pattern of IGF-1 by means of immunohistochemistry and Western-blot. We also used ELISA to measure the serum level of IGF-1.

RESULTS:(1)Result detected by immunohistochemistry: IGF-1 was expressed mainly in the cytoplasm of extraocular muscle cells, small amount of IGF-1 was expressed in the extracellular matrix. The expression of IGF-1 in extraocular muscle of normal eye position group was significantly more than that of concomitant esotropia group and concomitant exotropia group(P<0.05).(2)Result detected by Western blot: compared with concomitant esotropia group and concomitant exotropia group,the expression of IGF-1 in normal eye position group was significantly increased(P<0.05).(3)The difference of serum level of IGF-1 among normal eye position group, concomitant esotropia group and concomitant exotropia group was not statistically significant(P>0.05).

CONCLUSION: The occurrence of simple abnormal eye position such as concomitant esotropia and concomitant exotropia in children may has few association with the level of IGF-1 in serum, but be associated with the expression of IGF-1 in extraocular muscle. It suggests that injecting IGF-1 into extraocular muscle may be a potiential application for curing strabismus.]]> 2013/7/29 14:34:25 Li-Juan Tao,Yu-Lin Luo,Ping Wang,Xi-Lang Wang,Yan Guo and Yi-Lan Tan 281true METHODS: Sixty adult SD rats were randomly assigned into normal group(n=15), IR group(n=15), saline group(n=15), y-39983 group(n=15), and the rats in later three groups were induced retinal IR by intra-anterior chamber infusion of saline solution. Rats in the saline and y-39983 groups were intravitreously injected with normal saline and y-39983 respectively, 5 minutes prior to the induction of ischemia. The protein expression of intercellular cell adhesion molecules-1(ICAM-1)was determined by immunohistochemical staining. Retrograde labeling of retinal ganalion cells(RGCs)with fluorogold was performed to assess the number of RGCs. Histologic studies and electroretinogram were carried out to evaluate retinal damage.

RESULTS: The preadminisration of y-39983 decreased ICAM-1 expression, elevated the survival rate of RGCs and the recoverage rate of amplitude of b wave and O₂ wave significantly, and attenuated the IR-induced the thinning of inner retina.

CONCLUSION: y-39983 can protect the rat retina from IR injury at least partially by inhibiting ICAM-1 abnormal expression, suggesting that y-39983 may have therapeutic potential for the retinal diseases associated with IR.]]> 2013/7/1 11:07:54 Qing-Qing Meng and Su Liu 280true Pax6 gene in retinoblastoma(Rb).

METHODS: Totally 15 cases of fresh Rb organizations were selected as observation group and 15 normal retinal organizations as control group. Western-Blot and reverse transcriptase polymerase chain reaction(RT-PCR)methods were used to detect Pax6 protein and Pax6 mRNA expressions of the normal retina organizations and Rb organizations. At the same time, Western Blot method was used to detect the Pax6 gene downstream MATH5 and BRN3b differentiation gene protein level expression. After the comparison between two groups, the expression and clinical significance of Pax6 gene in Rb were discussed.

RESULTS: In the observation group, average value of mRNA expression of Pax6 gene was 0.99±0.03; average value of Pax6 gene protein expression was 2.07±0.15; average value of BRN3b protein expression was 0.195±0.016; average value of MATH5 protein expression was 0.190±0.031. They were significantly higher than the control group, and the differences were statistically significant(P<0.05).

CONCLUSION: Abnormal expression of Pax6 gene is likely to accelerate the occurrence of Rb.]]> 2013/7/1 11:07:54 Hai-Dong Huang,Li-Yang Li,Ying Guo,Xing Zhao,Jing-Jia Kang and Di Guan 279true in vitro cultured human corneal endothelial(HCE)cells and lay foundations for its secure medication clinically.

METHODS: After in vitro cultured HCE cells treated with timolol maleate at different concentrations, the situation of cellular growth, proliferation, and morphology was checked under an inverted light microscope. And plasma membrane permeability, DNA fragmentations, and ultra-structure of HCE cells were detected by acridine orange/ethidium bromide(AO/EB)double fluorescent staining, DNA agarose gel electrophoresis, and transmission electron microscopy(TEM), respectively.

RESULTS: After treated with timolol maleate at concentrations from 0.15625g/L to 5g/L, in vitro cultured HCE cells showed typical characteristics of apoptosis, including growth retarding, number decreasing, cytoplasmic shrinking, vacuolation, falling off from well bottom of culture plate, plasma membrane permeability increasing, chromatin condensation, DNA fragmentation, appearance of apoptotic body, and so on. The apoptosis-inducing effect of timolol maleate was in dose- and time-dependent manners. The greatest apoptosis-inducing effect of timolol maleate on HCE cells was found at the clinic medication concentration of 2.5-5g/L, and their induced apoptotic rate of HCE cells at 28 hours reached 83.23%-96.71%, respectively.

CONCLUSION: Timolol maleate within the concentration of 0.15625-5g/L has an obvious apoptosis-inducing effect on HCE cells, and has tremendous toxicity to HCE cells at clinic medication dosage. Timolol maleate should be carefully utilized in ophthalmic clinic.]]> 2013/6/3 10:33:25 Ting-Jun Fan,Bei-Bei Sui,Qing-Yang Wang,Qian Wen,Qian Sun,Miao-Miao Yu and Yuan Ge 278true METHODS:Subcultured human lens epithelial cell line, ultraviolet induced cell apoptosis, 20μmol/L resveratrol pretreated cell, the indicators change was observed: rate of apoptosis was detected by flow cytometry and apoptosis-related factors of caspses-3 and caspase-9 were detected by colorimetric detection, ultrastructure changes were observed under transmission electron microscope.

RESULTS: Flow cytometry instrument testing found that resveratrol can suppress the apoptosis induced by ultraviolet irradiation, caspses-3 and caspase-9 content in positive control group were significantly higher than that of the negative control group at the same time period, the difference was statistically significant(P<0.05); caspses-3 and caspase-9 content in experimental group were lower than that in the positive control group at the same time, the difference was statistically significant(P<0.05). In addition, the damage of human lens epithelial cells was alleviated with the incubation time of resveratrol elongated.

CONCLUSION:Resveratrol may inhibit ultraviolet-induced apoptosis of human lens epithelial cells, it has preventive function against radioactive cataract, and it can provide reliable evidence for pursuing effective medicine to prevent and treat cataract.]]> 2013/6/3 10:33:25 Xue-Fang Chen,Zhong-Xin Liu,Bing-Rong Chen,Ping Liu and Yi Zheng 277true METHODS: We induced diabetic rat model by peritoneal injection of STZ, after the blood glucose raised, we used Evans blue to trace the leakage of blood-retina barrier(BRB)every month. After blood glucose rose three months later, we treated the Chinese medicine group diabetic rat with Qidengmingmu capsule. There were three groups of different dose, low dose group of 125mg/kg, middle dose group of 250mg/kg, high dose group of 500mg/kg. The control group was treated with calcium dobesilate(200mg/kg). After three months treated by medicine, the leakage of rat blood-retina barrier was evaluated.

RESULTS: The damage of BRB and visual function occurred at two week after the blood glucose rose, and the damage aggravated with the continuing of high diabetic. But after the Chinese medicine treated three months, the rat's retina vessel leakage was reduced.

CONCLUSION: BRB break down and visual acuity damage appears in early phase of STZ diabetic rat and get worse as the hyperglycemia keep on. The Chinese medicine Qidengmingmu capsule can prevent the vessel leakage by damage of BRB.]]> 2013/6/3 10:33:25 Fu-Wen Zhang,Jun-Guo Duan,Ling Zhao,Xue-Jing Lu and Qiang Li 276true METHODS: One hundred and twenty male Sprague-Dawley(SD)rats(220±10g)were randomly divided into four groups(thirty in each group). αB group,(AOH following intravitreal injection of αB crystallin group, n=30). S group,(AOH following intravitreal injection of normal saline solution group, n=30). P group,(AOH following vitreous puncture group, n=30). H group,(AOH group, n=30). Western blot was assayed αB-crystallin content 7 and 14 days after model; Immunohistochemistry was observed GAP-43 expression 7, 14 and 21 days after model established; Amplitude of ERG b-wave was tested retinal function before and 1 month after model.

RESULTS: The αB content in retina of αB group was higher 7 days after model established, fewer 14 days after model(P=0.000, P=0.000, P=0.001, P=0.000). But the αB content in retina of αB group was more serious compared with other groups at the same time points(P=0.019,P=0.016); The GAP-43 expression of RGCs in αB group was highest 7 days after model established, started to decrease 14 days after model established, which was fewest 21 days after model established(P=0.000, P=0.000, P=0.000, P=0.000). But the GAP-43 expression of RGCs in αB group was significantly higher than in other groups at each time point(P =0.002, P =0.011, P =0.009); The amplitude of ERG b-wave was lower 1 month after model(P=0.014, P=0.004, P=0.003, P=0.006). There were no statistical differences among groups on amplitude of ERG b-wave before model established(P=0.993). The amplitude of ERG b-wave in αB group was higher than others 1 month after model(P=0.002).

CONCLUSION: Exogenous αB-crystallin could improve αB-crystallin expression of retinas; αB-crystallin contributed to the increase of GAP-43 expression of RGCs and RGCs axonal regeneration; αB-crystallin could promote the recovery of retinal function and was non-toxic on retinas of rat.]]> 2013/6/3 10:33:25 Rui-Hong Wang,Zhi-Hong Wu,Shi-Ke Hou,Hao-Jun Fan and Li-Hua Wang 275true METHODS: CNV was established in thirty SD rats by alkaline burning. Rats were divided equally to group A and group B at random. In group A, right eyes were experimental group A1, treated by 40μmol/L curcunmin solution, and left eyes were control group A2, treated by 0.09% sodium chloride. In group B, right eyes were experimental group group B1, treated by 5g/L avastin, and left eyes were control group B2, treated by 0.09% sodium chloride. Cornea and aqueous humor were collected by time spot. The capillary vessels were study, and the expressions of VEGF were detected by Enzyme-Linked immunosorbnent Assay(ELISA).

RESULTS: No toxic effects of the drugs were found. The capillary vessels in experimental group were less than those of control group(P<0.01). No statistical different of the capillary vessels between two drugs were found. The expressions of VEGF in experimental group were less than those in control group(P<0.01). The expressions of VEGF in B1 group were less than in group A1.

CONCLUSION: The inhibitory effect to CNV of curcunmin and avastin have no statistical different in the experiment, but curcunmin has the less inhibitory effect to the expressions of VEGF than avastin. Curcunmin may have other mechanism in the inhibitory action on CNV.]]> 2013/6/3 10:33:25 Yi Wu,Wen-Xiong Liao,Shou-Quan Lu,Jin-Sheng Li,Ai-Ping Gu and Hua Li 274true METHODS: Four effective components of Qing Guang An were used in rabbit eyes after glaucoma surgery(Group D.E.F.G), with the blank group(A), model group(B), MMC group comparison(C). Effects of four effective components of Qing Guang An on inoblast and typeⅠcollagen of scarring tissue of filtration canal after glaucoma surgery were observed.

RESULTS: After surgery, MMC group(C), effective component 2(E)groups of intraocular pressure rose slowly. At the fourth week, intraocular pressure remained minimum. The compare of these two groups of intraocular pressure and those of group A, B, D, F, G had significance on statistics. Comparing with the number of inoblast of each group, except group D and E, which result was P>0.05 and had no obvious significance on statistics, others were significant. Comparing with the expression of typeⅠcollagen, except group C and E, B and G, F and G, result of others had obvious significance on statistics showing as P<0.05.

CONCLUSION: The scaring of filter canal after glaucoma surgery is main cause of the filtration surgery's failure assuming abnormal backup intraocular pressure. The effective componment 2 of Qing Guang An and MMC can obviously inhibit the multiplication of inoblast and the expression of typeⅠcollagen, reduce the hyperplasia of scar tissue, which has obvious effect for preventing scarring of filtration canal.]]> 2013/5/6 14:25:10 Yan Liu and Qing-Hua Peng 273true 2)simulated hypoxic environment's effect on the gene expression and protein secretion of vascular endothelial growth factor(VEGF)of rats' retinal progenitor cells(RPCs).

METHODS: RPCs were separated, cultured and identified by the use of neurospheres-adhesive culture. RT-PCR and ELISA were used to detect the change of RPCs VEGF's expression after 24-hour intervention of CoCl₂ with different concentration(0, 50, 100, 150μmol/L and 200μmol/L), and detect the change of RPCs VEGF's expression in 150μmol/L CoCl₂ for different period(0, 12, 24, 36, 48, 72 hours).

RESULTS: The RPCs cultured by neurospheres-adhesive culture system could highly express the specificity of stem cells(Nestin), and express mature retinal neuron after natural differentiation. After culturing them in the hypoxic environment for the same period, the gene express of VEGF gradually increased and then decreased with the increasing concentration of CoCl₂, and peaked(P≤0.05)when the concentration of CoCl₂ was 150μmol/L. When the concentration of CoCl₂ was 150μmol/L, the gene expression of VEGF also gradually increased and then decreased with the increasing hypoxia intervention period, and peaked(P≤0.05)when the hypoxia intervention period was 36 hours, the change of protein and gene expression were consistent.

CONCLUSION: The RPCs cultured by neurospheres-adhesive culture system can better express the specificity of stem cells. Under CoCl₂ simulated hypoxic environment, hypoxia strengthens RPCs VEGF's secretion and dependent on the time and dose.]]> 2013/5/6 14:25:10 An-Ran Liang,Ming-Ying Lai and Ping-Hong Lai 272true METHODS:Totally 24 healthy New Zealand rabbits were randomly divided into 4 groups: normal control group(A group); high intraocular pressure model blank group(B group); high intraocular pressure model with GSTT treated group(C group); high intraocular pressure model with Erigeron brevicapas hand mass(EBHM)treated group(D group). High intraocular pressure model was induced by 20g/L methylcellulose injection into the anterior chamber in B group, C group and D group. D group was injected 5 mg/kg GSTT and C group was injected 4.5mg/kg EBHM and measured intraocular pressure with Schiotz tonometer every day for 4 weeks. The retina tissue superoxide dismutase(SOD)and maleic dialdehyde(MDA)content were detected 28 days later.

RESULTS: After glaucoma model of rabbit eyes were established, the intraocular pressure during observation period was maintained in 32-39mmHg; High intraocular pressure model blank group and normal control group, EBHM treatment group, GSTT treatment group were compared, the differences of retina MDA, SOD content had statistical significance(P<0.05); numerical difference between EBHM treatment group and GSTT group was not statistically significant(P>0.05); EBHM treatment group, GSTT treatment group and normal control group were compared, the content of MDA in the retina was still slightly higher(P<0.05), the content of SOD slightly lower(P<0.05)

CONCLUSION: GSTT can effectively improve the retina SOD activity of chronic high intraocular pressure in rabbit and reduce the content of MDA, which has a protective effect of persistent high intraocular retinal oxidative stress.]]> 2013/5/6 14:25:10 Nuo Li,Li-Na Huang and Ping Zeng 271true METHODS: Totally 26 patients who have been obtained epiretinal membranes(PDRⅤ-Ⅵ)were performed pars- plana vitrectomy. They were removed the epiretinal membrane intraoperation and to observe HIF-1α, VEGF and expression of vascular endothelial marker CD34 in epiretinal membranes through immunohistochemical method.

RESULTS:Vascular endothelial cells of PDR expressed HIF-1α in 24 cases(92.3%)and VEGF in 15 cases(57.7%)in membranes, respectively. There were significant correlations between the numbers of blood vessels expressing CD34 and HIF-1α(r=0.556, P=0.028). There were significant correlations between the numbers of blood vessels expressing CD34 and VEGF(r=0.745,P=0.001).

CONCLUSION:HIF-1α and VEGF may play an important role in the pathogenesis of PDR.]]> 2013/5/6 14:25:10 Jian-Feng Cao,Dong-Bo Pang and Wen-Jie Ye 270true METHODS: Twenty 8-week-old female NOD mice with dry eye caused by SS were randomly divided into 4 groups. One week later, the mice were treated with subconjunctival injection. GroupⅠ and Ⅱ received 200μg/kg and 400μg/kg rapamycin-loaded microspheres, Group Ⅲ and Ⅳ received normal saline and empty microspheres. Five 8-week-old female healthy KM mice were used as untreated controls. Before and 5, 10, 15, 20 days after the experiment, the amount of secretion of tears, the score of corneal fluorescein staining and rose bengal staining were investigated. Conjunctival epithelial cells were observed and graded by conjunctival impression cytology.

RESULTS: Compared with the group Ⅲ and Ⅳ, the amount of secretion of tears of the mice in groupⅠ and Ⅱ increased. The scores of corneal fluorescein staining and rose bengal staining were lower. The levels of conjunctival impression cytology reduced.

CONCLUSION: Rapamycin-loaded microspheres can decrease dry eye signs by alleviating the ocular surface inflammation of NOD mice. It suggests rapamycin-loaded microsphere is valuable to dry eye caused by SS.]]> 2013/5/6 0:00:00 Meng Wang and Yan Liu 269true METHODS: Primary culture and subculture of rabbit keratocytes were established in vitro. Plasmid vector pBos-Cite-neo/TFPI-2 was transfected into keratocytes with Lipofectamine 2000. After being selected by G418, three groups of cells including TFPI-2 gene transfected cells K-TFPI-2, empty vector transfected cells K-V and non-transfected cells K-P were screened for TFPI-2 mRNA and protein by reverse transcription-polymerase chain reaction and Western blot analysis, respectively. The activity of MMPs in the three groups of cells was detected by substrate zymography and compared by ANOVA.

RESULTS: Expression of mRNA and protein of TFPI-2 was more in the cells of K-TFPI-2 than in the other cells of K-P and K-V with a significant difference(mRNA:0.79±0.02 vs 0.51±0.03 and 0.48±0.02, P=0.000 and P=0.000; Protein:24.5±0.8 vs 15.5±0.5 and 14.9±0.9,P=0.000 and P=0.000). Compared with the two groups of K-P and K-V, the cells of K-TFPI-2 had a significant decreased activity of MMP1(12.3±0.7 vs 16.7±1.2 and 15.9±0.7, P=0.001 and P=0.003)and MMP2(15.4±1.3 vs 18.2±1.1 and 17.8±1.1, P=0.027 and P=0.046).

CONCLUSION: It is suggested that the expression of TFPI-2 may strongly inhibit the activity of MMPs in keratocytes in vitro, which provides an experimental basis for curing CNV with gene therapy.]]> 2013/4/7 15:27:38 Jing Yuan,Jian-Xiong Yu and Lian-Hong Zhou 268true METHODS:SD rats were given intraperitoneal injection of streptozotocin to induce diabetes. Ink perfusion and stretched retina were used to observe the retinal blood vessels. Immunohistochemistry staining was used to detect syndecan-1 protein in the rat retina and epiretinal membranes of proliferative diabetic retinopathy.

RESULTS:In the diabetic rats of 9-week, peripheral retinal blood vessels shaped tortuously, capillary network was significantly reduced, and parts of the capillary perfusion was bad. In the control group, strong positive staining of syndecan-1 was detected in the nerve fiber layer and ganglion cells, moderate positive staining was observed in the inner plexiform layer and outer segments of photoreceptor, and weak positive staining was detected in the outer plexiform layer. In the retina of diabetic rats, syndecan-1 decreased. Moderate positive staining of syndecan-1 was observed in the nerve fiber layer, ganglion cells and outer segments of photoreceptor, weak positive staining was detected in the inner plexiform layer and outer plexiform layer. Among 13 specimens of epiretinal membranes, weak positive staining of syndecan-1 was detected in 8 cases(61.5%)and negative staining was observed in 5 cases(38.5%).

CONCLUSION:Syndecan-1 is down-regulated in the retina of diabetic rats and epiretinal membranes of proliferative diabetic retinopathy.]]> 2013/4/7 15:27:38 Jing-Bo Wang and Huan-Long Yu 267true 2(TGF-β₂).

METHODS: Human RPE cells were exposed to different concentrations insulin at different times(0 to 48 hours). We used the MTS method to test the proliferation of RPE cells. And we used the ELISA method to text the secretion of TGF-β₂ of RPE cells in various cases, and the Real-time PCR method to test the expressions of TGF-β₂ mRNA.

RESULTS: The MTS results showed that after 12 hours exposure of insulin RPE cells proliferated significantly(P<0.05). ELISA results showed that insulin can significantly promote the secretion of TGF-β₂ of RPE cells, and this process was independent of time and dose. And Real-time PCR results showed that 24 hours after the stimulation of insulin, TGF-β₂ mRNA increased significantly(P<0.01), and 10×10³U/mL insulin group, TGF-β₂ mRNA expression were significantly higher than the 0.1× 10³U/mL insulin group(P<0.01).

CONCLUSION: RPE cells are important intraocular sources of TGF-β₂, and insulin can promote the proliferation of RPE cells and the secretion of TGF-β₂ to promote myopia.]]> 2013/4/7 15:27:38 Yue Zou,Meng-Zhu Wu,Feng Wang and Ying Fan 266true METHODS:Totally 130 eyes were obtained from freshly slaughter pigs and divided into 5 groups. Each group was sub-divided into the 15 minutes group and the 30 minutes group, with 13 eyes in each group. The control group were performed middle vitreous injection of 0.1mL PBS, 200U/mL and 800U/mL hyaluronidase(HA)were injected in groups A and B, 10U/mL and 50U/mL chondroitinase(CA)in groups C and D. After incubation for 15 minutes and 30 minutes, the eye balls were fixed with 4% glutaraldehyde and retina fixative solution. Pathological examination, eosin-hematoxylin staining, scan electron microscopy were taken to evaluate the remaining vitreous on vitreous base, transmission electron microscopy to evaluate retina toxicity.

RESULTS:Both of gross appearance and hematoxylin+eosine slice all revealed basal vitreous partially liquefied and degradation. Remarkably decrease was found in basal vitreous than the control group both in the HA 800U/mL group and the CA 50U/mL group when using electron microscopy scanning. Transmission electron microscopy in group B, C and D revealed the remaining vitreous were less than the control group.

CONCLUSION: Both CA and HA can cause basal vitreoretinal detachment in enucleated pig eyes. But the HA may cause less damage to the retina.]]> 2013/4/7 15:27:38 Yi-Bin Xiong 265true 2O₂-induced lens epithelial cells.

METHODS: SD rats transparent crystals were cultured in vitro by lens technology, and placed in a certain concentration of H₂O₂ in MEM medium. The experimental cataract lens model was established. Blank control group, H₂O₂ group and tea polyphenols(0.02mg/mL, 0.2mg/mL, 2mg/mL)treatment group were set and sampled at different time points(6, 12, 24, 48 hours). NF-κB mRNA expression was detected by RT-PCR, and NF-κB protein expression was detected by Western-blot in lens epithelial cells.

RESULTS: H₂O₂ raised the expression of NF-κB, and appeared time-dependent manner(P<0.01)in lens epithelial cells; different concentrations of tea polyphenols can inhibit the expression of NF-κB in lens epithelial cells, when TP concentration was 0.2mg/mL, the inhibitory effects was statistically significant(P<0.05).

CONCLUSION: TP can inhibit the expression of NF-κB in H₂O₂-induced lens epithelial cells, when TP concentration was 0.2mg/mL, the effect is statistically significant.]]> Bao-Cheng Men,Shu Zhang and Dan Liu 264true METHODS: The animal model of retinal injury was created by a free falling iron bar hitting the rat cornea. The rats were divided into normal group(n=6), contusion group(n=24)and treatment groups of NAC(n=24)randomly. After retina contusion, the latter two groups were subdivided into 1 day, 3 days, 7 days and 14 days group; there were 6 rats in each observation period. Structural changes of retina were observed with light microscope, and the expression of Bcl-2/Bax was observed by immunohistochemistry.

RESULTS: In the contusion of the rat retina, retinal damage was concentrated in sensory layer. RGC layer, inner nuclear layer, and outer nuclear layer cells were obviously observed. After contusion, retinal tissue edema, cell disorder, cytoplasm vacuolated changes, thinning of retinal tissue, and cell loss were began to appear. No expression of Bax was found in normal group and treatment group, and low expression of Bcl-2 was observed. Most of Bax positive cells had expression at 1 day after contusion injury in contusion group, and then increased obviously at 3 days, and decreased at 7 days and much further at 14 days, and Bcl-2 positive cells had expression at a low stage. Treatment group had the same trend with the contusion groups in each index, but the expressive intensity of each period weakened in evidence. There were significant differences between two groups at 1 day,3 days and 7 days after contusion(P<0.05). The expression of Bcl-2 in each period of time was increased. Compared the two groups with 1 day, 3 days after contusion and 7 days had a significant difference(P<0.01 ).

CONCLUSION:In the contusion of the rat retina, NAC can improve the retina damage, and may protect the retinal tissue from contusion injury by regulating Bcl-2/Bax protein expression.]]> Jing-Jing Liu and Dan Liu 263true 2+ of guinea pig retinal pigment epithelial(RPE)cells in vitro and compare the changes of Ca²⁺ with or without Ver under different forms of light.

METHODS: Ten two-week-old healthy guinea pigs were chosen and RPE cells were cultured in vitro. Cells were divided into Ver treated and untreated groups, then each group further divided into 4 groups: focused light group, defocused light group, parallel light group and control group. The first three groups were exposed to the focused light, defocused light(2 forms of light were transformed from parallel light by passing through different lens)and the parallel light respectively, and control group was removed from exposure of light. After exposed to different forms of light, the fluorescence intensity of intracellular Ca²⁺ were detected by laser scanning confocal microscope(LSCM)immediately. Cool white light was used as light source. Cells were exposed to light with same spot diameter and degree of irradiation level(at 300 LUX)for 30 minutes respectively. Horizontal temperature of cells changed between 36.5℃-37.2℃. There was no natural light interference. In order to avoid the impaction of refraction of liquid, most of the medium were siphoned off before irradiation. The data were analysed by SPSS 13.0 statistical software, completely randomized design ANOVA and Pearson linear correlation analysis were used as statistical methods. The forms-effect relations were explored.

RESULTS: Treated with Ver for 12 hours, the apoptosis rates of RPE cells in 20, 40, 80mg/L concentration group had no significant difference compared with control group(P>0.05). Ver could reduce the Ca²⁺ fluorescence intensity of RPE cell,and there was significantly statistical difference in 80mg/L group(P<0.05). The Ca²⁺ fluorescence intensity of focused group of no Ver treated part was significantly higher than other groups, had significant differences compared with the other groups(P<0.05). After added 80mg/L Ver on the RPE cells for 12 hours, exposure to light did not significantly increase the Ca²⁺ fluorescence intensity, there was no significant difference among the 4 groups(P >0.05).

CONCLUSION: Focused light can significantly stimulate the concentration of intracellular Ca²⁺ of RPE cells. Ver above a certain concentration can induce apoptosis of RPE cells; 80mg/L Ver can effectively reduce intracellular Ca²⁺ fluorescence intensity under the premise of not leading to apoptosis of RPE cells. Different forms of light have no effect on the Ca²⁺ fluorescence intensity of RPE cells after treated with Ver(80mg/L)for 12 hours.]]> Jiang-Yuan Qin and Chao-Ying Wang 262true METHODS: Rat corneal alkaline burn model was established in both eyes by routine method in 40 anesthetized female Sprague-Dawley rats. The rats were randomly assigned to 2 groups with 20 rats each for topical administration of recombinant PEDF combined with chloramphenicol or normal saline combined with chloramphenicol(as control). At different intervals(4, 7, 10, and 14 days)of the treatment, rats were euthanized and the corneas removed for immunohistochemistry analyses to measure expression levels of PEDF, vascular endothelial growth factor(VEGF), Fas and FasL. The eyes of ten healthy rats were used as normal control. Meanwhile, the apoptosis of neovascular endothelial cells was detected by TUNEL method on the 4^th, 7^th, 10^th and 14^th day respectively after the burn.

RESULTS: There were high levels of PEDF expression and low levels of VEGF, Fas and FasL in the normal cornea. VEGF levels were significantly induced by chemical cauterization in the groups treated with chloramphenicol combined with normal saline, demonstrating CNV. In contrast, the PEDF treatment prevented the over expression of VEGF induced by the cauterization and unregulated the expression of Fas and FasL. In CNV tissues, the positive immune reaction of VEGF was most apparent during the 7^th day and then declined thereafter. However, the most positive expression of PEDF, Fas, FasL was on the 10^th day and then declined slowly after thereafter. The ratio of PEDF/VEGF raised from <1 to >1 in the course and the hinge was on the 10^th day. Certain time correlation existed between the dynamic expressions of VEGF and PEDF and the development of CNV. The expression of Fas and FasL correlated to PEDF closely in the whole procession which may underlie a simulative relationship between them. The apoptosis of CNV endothelial cells expressed most positively on the 10^th day and it was always much more intense in the PEDF group than in the normal saline group.

CONCLUSION: The expressions of VEGF and PEDF are remarkably expressed in experimental rat CNV tissues, and the fluctuation of which is consistent with the development of CNV. The breakdown of the balance between the two factors may play a role in CNV occurrence and development. That the PEDF downregulates VEGF expression and upregulates Fas and FasL expression which induces the apoptosis of CNV endothelial cells results in the inhibition of corneal NV induced by chemical cauterization. The results suggested the possible mechanism of PEDF in the therapeutic function for CNV diseases.]]> Yue Fu,Xiao-He Lu,Dan Zhu,Min Fu and Wei Wu 261true METHODS: Thirty eyes of 30 SD rats were chemically cauterized by applying a 3mm-diameter filter paper soaked 1mol/L NaOH solution at the central cornea for 40 seconds. All animals were randomly assigned to five groups: group A, the PEDF drops was used the day after cauterization; group B, the PEDF drops was used the third day after cauterization; group C, the Avastin drops was used the day after cauterization; group D, the Avastin drops was used the third day after cauterization; group E, normal saline solution drops was used the day after cauterization(control group). All the eye drops were applied 10μL every time and 4 times per day. The growth of CNV was observed by slit lamp microscope and the area of CNV was calculated. Expression of VEGF and CD31 was detected by immunohistochemistry on the twelfth day after experiment.

RESULTS: On the twelfth day, the area of CNV in group A was smaller than that of group B, C(P<0.01). There was no significant difference in terms of CNV areas between group B and D(P=0.215)and between group C and D(P=0.058). Immunohistochemistry revealed that the expression of VEGF and CD31 was little in group A and massive in group E. In group B, C and D, expression of VEGF and CD31 was more than that of group A.

CONCLUSION: Topical application of PEDF and Avastin can inhibit alkali-induced CNV, moreover PEDF is superior to Avastin if applied early after cauterization.]]> Dan Zhu,Xiao-He Lu,Yue Fu,Wei Wu and Xiao-Hong Chen 260true METHODS: Wistar rats were randomly divided into three groups: control group, diabetic group and 3-AB group. The rats of diabetic group and 3-AB group were treated with intraperitoneal injection of streptozotocin(STZ). The rats of control group were given the same volume of citrate buffer. After the model constructed, 3-AB group was treated with 3-AB(30 mg/kg)gavage daily while the control group and diabetic group were given the same volume of 0.9% NS instead. The progress of lens opacity of rats was observed and recorded. After administrated for 2, 4 and 8 weeks, rats were sacrificed and taken lens respectively. The lens were used to detected the activity of glutathione peroxidase(GSH-px), superoxide dismutase(SOD), the level of malondialdehyde(MDA)and advanced glycation end products(AGE), the expression of matrix metalloproteinase-2(MMP-2)and basic fibroblast growth factor(bFGF)in lens epithelial cells.

RESULTS: Lens of diabetic group and 3-AB group appeared various degree of cloudy since the third week. The level of diabetic group was higher than 3-AB group(P<0.01)in the fourth and eighth week. The GSH-Px and SOD activities of diabetic group were lower than 3-AB group(all P<0.05)at the 2, 4, 8 weeks, but all of them lower than control group(P<0.01). The content of MDA increased in diabetic group and 3-AB group, compared with control group(P<0.01), and its content in diabetic group was higher than in 3-AB group(P<0.05)at the 2, 4, 8 weeks. The content of AGE in the lens that come from control group were lower than diabetic group and 3-AB group(all P<0.05)in each time. At 2, 4 weeks, its content in diabetic group was higher than in 3-AB group(P<0.05), but there was no difference at 8 weeks(P>0.05). There was rare expression of MMP-2 in control group, the MMP-2 expression of diabetic group is statistically significant higher than 3-AB group(P<0.05)in the 2, 4, 8 weeks. bFGF was expressed in the three groups, and the bFGF expression of control group was lower than diabetic group and 3-AB group(all P<0.05)in the 2, 4, 8 weeks. There was no difference between diabetic group and 3-AB group in each time.

CONCLUSION: 3-AB has a certain inhibitory effect for lens opacity in the STZ-induced diabetic rats, the possible mechanism: alleviates the oxidative damage of lens epithelial cells, reduces the level of non-enzyme glycation, and inhibits the expression of MMP-2 suggested that it can alleviate the degradation of lens epithelial cells' extracellular matrix, which lead to inhibit the lens opacity.]]> Jun-Ling Wang,Gang-Jin Kang,Dong Qin,Lin Mou and Mei Xu 259true METHODS: Totally 60 neonatal rats were randomly allocated into 6 groups: normal animals(Nor), normal animals treated with PBS(Nor+PBS)or NEP1-40(Nor+NEP), MD animals without any treatment(MD), MD animals treated with PBS(MD + PBS)or NEP1-40(MD + NEP). We subjected P21 rats to monocular deprivation model until P45. Then the deprived eyes of MD model rats were reopened and followed by NEP1-40 or PBS administration for 7 days. Ultrastructral modifications of synaptic junctions and objective visual function were examined at P52 to determine the therapeutic effects of NEP1-40.

RESULTS: The objective visual function examined by F-VEP: At P45, F-VEP showed that compared with rats in Nor group, deprived eyes of rats in MD group, MD+PBS group and MD+NEP group displayed a longer latency(P<0.05)and a smaller amplitude(P<0.05); At P52, we could see that comparing with the MD group and MD + PBS group, the latencies of F-VEP in MD+NEP group were shortened and the amplitudes were increased(P<0.05), which were similar with that of in Nor group(Nor vs MD+NEP, P>0.05). The structural modifications of synaptic junctions examined by electromicrographs: All of the structural parameters of the synaptic junction in visual cortex were altered by monocular deprivation in MD group compared with the Nor group, displaying an increased width of synaptic clefts, shortened synaptic active zone, decreased curvature of synaptic interface and decreased thickness of PSD. However, synaptic ultrastructural analysis showed that NEP1-40 treatment could recover the entire structural index in monocular deprivation rats(MD vs MD+PBS vs MD+NEP, P<0.05)but not normal rats(Nor vs Nor+PBS vs Nor+NEP, P>0.05). It was noteworthy that, although the width of synaptic clefts in MD+NEP group decreased remarkably in comparison with that of in MD group, it still had not reach the normal level(Nor vs MD+NEP, P>0.05).

CONCLUSION: NEP1-40 could recover the structural modifications of synaptic junctions of neurons in visual cortex contralateral to deprived eye, as well as the objective visual function of deprived eye, which indicated a new role for NEP1-40 in reactivation of visual cortical plasticity from monocular deprivation in adult rats and offered the theoretical foundation for curing adult patients with amblyopia in the clinic.]]> Yu-Lin Luo,Xiao-Ying Wu,Shuang-Zhen Liu and Li-Juan Tao 258true in vitro under the condition of high glucose.

METHODS: Human retinal capillary endothelial cells(HRCECs)extraction was taken from fresh human eyeand cultured in vitro. The cells of good growth in 3^rd-4^th generation were used into the experiment. The experimental subjects were divided into low glucose control group, high glucose control group and high glucose+different concentrations ART(15μg/mL, 30μg/mL, 60μg/mL, and 120μg/mL)groups. The effect of ART on the proliferation of HRCECs for 24 and 48 hours was measured by MTT assay. The effect of ART on the expression of VEGF in HRCECs was detected by immunocytochemistry.

RESULTS: The results from MTT assay showed that high glucose significantly increased the proliferation of HRCECs(P>0.05)and treatment of HRCECs with ART at the concentrations 15μg/mL,30μg/mL,60μg/mL,120μg/mL for 24 and 48 hours caused a time- and dose-dependent inhibition in the proliferation of HRCECs(P<0.05). The expression of VEGF in high glucose group was significantly higher than that in the low glucose control group(P<0.05). Compared with the high glucose control group, the expressions of VEGF in high glucose + different concentrations ART(15μg/mL,30μg/mL,60μg/mL)groups were decreased in a dose-dependent manner(P<0.05).

CONCLUSION: ART concentration-dependently inhibits high glucose-induced proliferation and VEGF overexpression in HRCECs, indicating that ART-inhibited the proliferation of HRCECs exerted by high glucose environment might be related to the inhibition of VEGF expression.]]> Yi Liu,Hui-Can Peng and Hong Tang 257true METHODS: The expression of Cav-1 was examined by immunofluorescence combined with confocal microscopy. Retinal structure in wild type(WT)and Cav-1 null mice was examined by HE staining. In vivo retinal function in WT and Cav-1 null mice was assessed by full-field scotopic electroretinography(ERG)following overnight dark adaptation.

RESULTS: Cav-1 null mice displayed reduced a-wave and b-wave amplitudes and reduced sensitivity as measured by ERG. However, the general structure of Cav^-/- retina was largely normal. Collectively, this implies the impairment of retinal function in Cav^-/- retina was not intrinsic to photoreceptor.

CONCLUSION: The observation that Cav-1 null mice show abnormal retinal function, in vivo but normal photoreceptor structure suggests that the retinal microenvironment rather than the photoreceptor itself is impaired. These may result in a disturbance in retinal pH, water or ion homeostasis therefore altering the subretinal milieu.]]> Yang Liu,Hong-Tao Xu and Xiao-Man Li 256true METHODS: EIU model was established by injecting a dose of 1mg/kg endotoxin(LPS)into the footpads of SD rats. At different time points(6, 12, 18, 24, 48 hours)after endotoxin injection, clinical symptoms were observed with ophthalmoscope. The cells in the aqueous humor one eye in each rat were counted. Eyes were enucleated for histological examination at different time points. Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay(TUNEL)method was used to assess the infiltrated cells apoptosis. The expression of TRAIL was determined by sreptavidin-biotin complex(SABC)immune histochemistry. The positive stain section was analyzed by computer-based image analysis system.

RESULTS: Intraocular inflammation appeared 6 hours after LPS injection, peaked at 18-24 hours, and obviously weakened at 48 hours. The cells of aqueous humor were gradually increased peaked at 24 hours. TUNEL stain indicated: a number of apoptotic infiltrated cells could be seen at the time of 6 hours to 24 hours after endotoxin injection. TRAIL was constitutively expressed on the rat iris weakly. TRAIL was expressed on the infiltrated cells. The intensity was weak in the 6 hours, 12 hours, at a peak in the 18 to 24 hours.

CONCLUSION: Uveitis was induced in the rat eyes with injecting 1mg/kg endotoxin. The short duration of EIU may be associated with apoptosis of infiltrated cells in inflammation. The TRAIL system may be associated with the infiltrated cells apoptosis in EIU.]]> Hui Man,Xu-Dong Huang and Jing Huang 255true METHODS: Totally 40 neonatal rats were randomly allocated into 6 groups: normal group of postpartum 28 days(NorP28), normal group of postpartum 35 days(NorP35), normal group of postpartum 42 days(NorP42), monocular deprivation group of postpartum 28 days(MDP28), monocular deprivation group of postpartum 35 days(MDP35)and monocular deprivation group of postpartum 42 days(MDP42). For the MD model, we sutured together the right eyelids of rats at postpartum 21 days. A subset of rats at each time points were killed, then the retinas were removed and frozen for detecting the expression pattern of BDNF and its receptor TrkB by means of Western blot and immunohistochemistry.

RESULTS: During the visual development critical period, the expression of BDNF and TrkB receptor had no significant change with the growth of age. After the early monocular deprivation, BDNF and TrkB receptor were significantly increased its expression at postpartum 28 days, then decreased at postpartum 35 days and maintained a steady level between postpartum 35 days to postpartum 42 days. BDNF expressed in ganglion cell layer, inner nuclear layer and outer nuclear layer of retina, while TrkB receptor expressed only in ganglion cell layer. In response to MD, the expression of BDNF and TrkB receptor positive cells was dramatically up-regulated in ganglion cell layer at postpartum 28 days, and then decreased gradually at postpartum 35 days to postpartum 42 days.

CONCLUSION: The expression of BDNF and its TrkB receptor presents a visual-experience dependency, and they participate in the onset of monocular deprivation amblyopia.]]> Yu-Lin Luo,Jun Luo,Zi-Feng Deng,Yan Guo,Xi-Lang Wang,Yi-Lan Tan,Li-Juan Tao and Xiu-Ting Wu 254true in vitro conditions of high glucose.

METHODS: The extracted from corneal transplantation HRSEC and cultured in vitro, which being good growth and at the 3- 4^th generation were taken into experiment. The experimental were divided into blank control group, high glucose control, low glucose control group, high glucose control group,high glucose+different concentrations of SF(1mg/L,2mg/L,4mg/L)group. The result of SF with different concentrations on the proliferation of HRCEC for 48 hours was surveyed by MTT assay. The immunocytochemical method was used to detect low glucose control group, high glucose group and different concentrations of SF(1mg/L, 2mg/L, 4mg/L)group HRCEC of VEGF expression.

RESULTS: The results of MTT colorimetric method showed that: different concentrations of SF in primary cultured HRCEC for 48 hours, cell proliferation inhibition rates were 46.97%, 61.55%, 76.91% and 83.47%. Within a certain range of concentrations of SF can restrain the proliferation of HRCEC in high glucose environment(P<0.05), and in 48 hours was concentration dependent. Low glucose control group compared with the high glucose control group the differences were not statistically significant(P=0.067>0.05). Low glucose control group, high glucose group and SF in concentrations of 1mg/L, 2mg/L, 4mg/L in HRCEC for 48 hours, immunocytochemistry of background gray values with positive gray value difference were respectively 28.27±1.62, 93.67±0.81, 72.67±2.89, 53.73±1.70, 30.93±3.72. Compared to the high glucose control group, expression of VEGF was decreased significantly in experimental groups after 48 hours, the difference was statically significant(P<0.05), in a dose-dependent manner. Compared to the low sugar glucose, VEGF in high glucose control group expression was significantly increased(P<0.05).

CONCLUSION: SF can reduce the HRCEC proliferation and suppress the expression of VEGF in high glucose environment which were cultured in vitro.]]> Qi-Lin Cheng,Hong Li,Hong Tang and Hui-Can Peng 253true METHODS:Mouse model of hyperoxia-induced ischemic retinopathy were established. A novel flucorescein-dextran perfusion method was developed to assess the vascular pattern. Histologic methods were used to count blood vessel profiles in the inner retina with HE staining. Survivin and COX-2 expressions in the retina were determined by immunohistochemical method.

RESULTS: Lots of neovascularization were seen in hyperoxia group. The number of nuclei of proliferative retinal vessels increased significantly as compared with normal control group(P<0.01). The expression of Survivin was noted in the ganglion cells and the neovascularization breaking through the inner retinal. The expression of COX-2 was noted in the inner nuclear layer and ganglion cells and retinal blood vessels. The expressions of them have positive correlation(r=0.394,P<0.01).

CONCLUSION:There are high expressions of Survivin and COX-2 protein in the retinal neovascularization and they have correlation.]]> Ning-Ning Liu,Ning Zhao,Li-Min Liu,Lei Chen and Na Cai 252true METHODS:Thirty healthy adult New Zealand white rabbits were randomly divided into experimental group and control group. Ultrasonic phacoemulsifications were performed in the 25 experiment rabbits under intramuscular injection anesthesia. The rabbits'eyes were examined by slit lamp microscope to observe the development of PCO after surgery and at 3d, 7d, 14d and 28d. The 5 rabbits in control group were executed to take the posterior capsule of right eyes. Reverse transcription polymerase(RT-PCR)and western-blotting were performed to detect the expression of Livin in PCO at different time points postoperatively.

RESULTS: Both RT-PCR and western-blotting method indicated that different levels of expression of Livin could be detected in the tissue of PCO in group B, C, D, E except control group and group A that instantly after surgery. The two methods indicated that Livin reached the peak in group C and decreased in group D, lower in group E and in group B reached the bottom.

CONCLUSION:The expression of Livin can be detected in the tissue of PCO in a certain similar time. The study indicated that Livin correlates with the pathogenesis of PCO. It may provide a novel tool for the investigation of gene therapy for PCO.]]> 2013/11/25 11:01:51 Shu-Jun Liu,Gui-Qiu Zhao,Yuan-Bin Li,Xue-Jing Man,Wen-Ting Wang and Zhen-Hua Zhang 251true METHODS: Cataract lens samples were dissolved in lysis buffer and sonication; the samples were extracted by TCA/acetone and protein extraction kit after centrifugation. Lens proteins of the samples were analyzed by two-dimensional electrophoresis(2-DE), gel electrophoresis and staining, image acquisition and image analysis.

RESULTS: According to the results obtained from two-dimensional electrophoresis gel image, the molecular weight of protein points were localized at 14-97.4kDa, PI5-9. The molecular weight of high abundance crystallins were localized at 20-31kDa, the molecular weight of lower abundance crystallins were localized at 35-45kDa. The 2-DE image background of cataract lens sample with hyaluronic acid extracted by TCA /acetone was clear, no obvious vertical bands, point of proteins were more clear, and 35 to 40 protein points were detected.

CONCLUSION: TCA/acetone extraction method has better purification compared to other methods in human cataract lens proteomics, and provides the reliable two-dimensional polyacrylamide gel Image for human cataract lens proteomics mass spectrometry analysis.]]> 2013/11/25 11:01:51 Zhao-Dong Chu,Guo-Hua Lu,Ying Tan,Yi Liu and Chao-Wei Li 250true in vitro and to investigate the possible mechanism of lycium barbarum polysaccharide(LBP)protecting the hRPE cells from blue light irradiation-induced damage.

METHODS: hRPE cells were cultured in Dulbecco's modified eagle's medium with high glucose and were pretreated with different concentrations(0.01mg/mL, 0.1mg/mL and 1mg/mL, respectively)of LBP solution, respectively. Further, hRPE cells were irradiated with different intensities(2000±500 LUX)of blue light(wave length 470-520nm)for 12h and the cellular morphology was observed by inverted phase contrast microscopy for every sample. Using flow cytometry, the alterations in the levels of reactive oxygen species, mitochondrial membrane potential and apoptosis were determined, respectively.

RESULTS: The level of reactive oxygen species in hRPE cells in blue-light treatment group was the highest, whereas the level in control group was the lowest. The fluorescent level of reactive oxygen species among different concentrations of LBP-treatment groups had specifically significant differences compared with that in blue light irradiation group(P<0.05). The amounts of apoptotic cells in different concentrations of LBP-treatment groups were specifically significant differences compared with that in blue light irradiation group(P<0.05); There were no apparent significant difference between 1mg/mL LBP-treatment group and normal control group(P>0.05).

CONCLUSION: LBP can efficiently inhibit apoptosis induced by blue light irradiation in hRPE cells and 1mg/mL of LBP has the strongest protective ability in blue light irradiation-induced damage in hRPE cells. The possible mechanism may attribute to the protective effect of LBP in inhibiting the overgeneration of reactive oxygen species and apoptosis in hRPE cells.]]> 2013/11/25 11:01:51 Wei-Hong Dong,Xiu-Juan Du,Da-Dong Guo,Ran Dou,Xiao-Feng Xie and Hong-Sheng Bi 249true METHODS: Twenty healthy rabbits were randomly divided into two groups: 16 in group A(laser group)and 4 in group B(normal control group). According to different time after laser photocoagulation rabbits were divided into four subgroups: 1d, 1 week, 2 weeks, 1month, group A with 4 for each subgroup; group B with 1 for each subgroup. To observe the histopathologic changes we used fundus photograph, light microscope, electron microscope and spot diameter calculating in different time points.

RESULTS:(1)Fundus photography: the spot region changed from thick white edema appearance on 1d after the surgery to melanoma attachment by 1 month;(2)Light microscopy: the main damage spot region was concentrated in the outer retina after photocoagulation. Photoreceptor cell appeared necrosis and apoptosis. With the passage of time, glial cells, pigment cells filled injury regional, and fibrous proliferation was formed;(3)Electron microscope: after photocoagulation, structure of membranous disc blurred and arranged in disorder; mitochondrial cristae became vague, fracture or even disappear; obvious vacuolization appeared. It began to appear apoptosis after 1 week, and gradually the fibroblast proliferation appeared;(4)Spot size: the spot size was the maximum at 1d, then gradually became smaller, spot size down to 85% at one-week time, 80% at 2-week time, and 79% at one-month time.

CONCLUSION: Frequency-doubled 532nm laser irradiation of pigmented macular, spot reaction is most obvious at 1d and stabilized at about 2 weeks.]]> 2013/11/25 11:01:51 Rui Guo,Long-Shu Shen and Yu-Liang Wang 248true MEATHODS: A total of 150 Wistar male rats of 1 month old, weighted about 200g, were randomly divided into 5 groups with 30 rats in each group with A representing normal group; B representing sham operation group; C representing surgery control group; D representing group treated with androgen; E representing group treated with buddleja officinalis total flavonoids. For the groups C, D, E, the bilateral testicle and epididymis were excised; For group B, scrota were incised without removal of the testicles, as the sham operation group; For group A, nothing was done. One week after modeling when the wound was to be healed, drug was given to each group. Respectively at the 1^st month, 3^rd, and 5^th months after treatment, 10 rats were randomly selected in each group, to receive Schirmer I test, tear breakup time measurement. Blood serum testosterone levels were tested in the fifth month.

RESUITS: For groups D and E, the Schirmer I test measurements were significantly higher than that of group C(P<0.05), tear film breakup time was significantly longer than that of group C(P<0.05). Serum testosterone levels in groups C and E were significantly lower than that in groups A, B, D(P<0.01).

CONCLUSION: Decreased androgen levels can lead to xeroma, and removal of bilateral testes and epididymis can successfully establish the animal models of xeroma in rats caused by decreased androgen levels. Buddleja officinalis total flavonoids have androgenic effect, which produces the similar treatment effect of xeroma with testosterone propionate. Buddleja officinalis total flavonoids may become a new treatment for xeroma.]]> 2013/10/28 11:29:57 Hai-Zhong Li,Qing-Hua Peng,Fen Wang,Xiao-Lei Yao and Wen-Juan Li 247true METHODS: Totally 60 BN rats were randomly divided into 4 groups: normal control group, model group, experimental group, physiological saline group with 15 in each group. All CNV models were made by krypton laser. Rats in experimental group were intraperitoneally injected with 0.35% EGb761(100mg/kg)every day after laser exposure until they were sacrificed. Rats in physiological saline group were intraperitoneally injected physiological saline every day after laser exposure until they were sacrificed. Fundus fluorescein angiography(FFA)was performed on every rat on the 7th day, 14th day and the 21st day after laser exposure, then the rats were sacrificed immediately. The eyes were enucleated and processed for histopathologic examination.

RESULTS: There was no choroidal fluorescein leakage staining in normal rats. There were obviously less choroidal fluorescein leakage points in experimental groups than that in the corresponding model groups(P<0.05), the difference had statistical significance. The structures of the retina and choroid were preserved better in experimental group than that in the model group.

CONCLUSION: EGb761 len inhibit the formation of laser-induced CNV in rats. The longer the time, the better curative effect.]]> 2013/10/28 11:29:57 Ling Wang,Chao Chen,Jun Liu and Hong-Sheng Bi 246true 1(TGF-β₁)on gene expression of main components of Smads family including Smad3, Smad4 and Smad7 in orbital fibroblasts(OF)of fibrosis extraocular muscle with thyroid -associated ophthalmopathy(TAO).

METHODS: OF were treated with 5μg/L TGF-β₁ at different time points(0min, 15min, 30min,1h, 2h and 4h), and real-time quantitative RT-PCR was performed to observe the effects of TGF-β₁ on the expression of Smad3, Smad4 and Smad7 mRNA.

RESULTS: The expression of Smad3 mRNA was 10.71 times to that of control group at 15min and 25.07 times at 1h(P<0.01); Smad4 mRNA was 1.54 times to that of control group at 15min and 15.99 times at 1h(P<0.01); Smad7 mRNA was 3.21 times to that of control group at 30min and 14.66 times at 4h(P<0.01).

CONCLUSION: TGF-β₁ up-regulate the expression of Smad3, Smad4, Smad7 mRNA in OF in a time dependent fashion, TGF-β₁/Smad pathway may play an important role in the pathogenesis of extraocular muscle fibrosis with TAO.]]> 2013/9/23 14:08:04 Yan Lu,Ying Ding,Pei-Li Hou,Hu Meng,Yu-Hua Shi and Zhen-Ping Huang 245true METHODS: RT-PCR and Western-Blot tests were carried out. In RT-PCR test, 60 adult SD rats were divided into normal group, antagonist group, and diabetes group. After diabetic rat model was induced using streptozotocin and antagonist group and diabetic group were injected intravitreously and postocularly with AMD3100 and PBS respectively. All rats were killed and the retina was extracted. after 1,3,5 months and the HE stain of paraffin sections was used and the expression of SDF-1 and VEGF mRNA were measured with RT-PCR. In Western-Blot test, 18 rats were divided into normal group, diabetes group and four antagonist groups which were using different concentration of AMD3100, and killed after 3 months.

RESULTS: SDF-1 and VEGF mRNA were expressed in normal group, antagonist group and diabetes group. At the same age group(1, 3 and 5 months)and among the normal group, antagonist group and diabetes group, the difference of expression of SDF-1 and VEGF mRNA were significant. The expressions in diabetic group were always highest and antagonist group lower than diabetic group. The expression of SDF-1 and VEGF mRNA was increased significantly with the extension of disease. The HE Stain of paraffin sections showed DM group had more cell nucleus which protruded internal limited membranes than normal control group and antagonist group. The Western-Blot test showed in 4 antagonist groups the SDF -1 and VEGF protein expression levels gradually decreased with the increases of SDF-1 antagonist AMD3100 concentration, the difference was significant. When intravitreous injected concentration of AMD3100 increased over 10μg/μL, the expression of SDF-1 and VEGF protein did not change, the difference was not statistically significant

CONCLUSION: With the progression of diabetic retinopathy, the expression of VEGF and SDF-1 mRNA in the retinal tissue of diabetic rats increased. The antagonist AMD3100 could reduce the expression of SDF-l, VEGF and inhibit the development of new blood vessels. In a certain concentration range, this inhibitory effect of AMD3100 was dose-dependent.]]> 2013/9/23 14:08:04 Yi-An You,Zhong-Yan Lai and Qian-Qian Zhou 244true METHODS: We cultured HRCEC both in normal and high glucose conditions. Each condition we treated the cells with 0.1mmoL/L ginsenoside Rg3 and 0.5mmoL/L ginsenoside Rg3. The effect of ginsenoside Rg3 on HRCEC proliferation was tested by methylthiazoletrazolium(MTT)assay 24h, 48h and 72h after the treatment; The effect of cell migration was tested by transwell; The effect of tube formation was tested by Matrigel; Western-Blot and real-time quantitative RT-PCR were used to detect the expression of vascular endothelial growth factor(VEGF)protein and mRNA. Data were statistically analyzed by ANOVA method.

RESULTS: Ginsenoside Rg3 could inhibit proliferation, migration and tube formation of HRCEC in both conditions, depending on the concentration and time. Ginsenoside Rg3 could decrease the expression of VEGF mRNA and protein in both conditions.

CONCLUSION: Ginsenoside Rg3 could inhibit VEGF expression, thus suppress the proliferation, migration, and tube formation of HRCEC in normal and high glucose conditions.]]> 2013/9/23 14:08:04 Liang Cao,Yu Song,Ying Wu and Li-Li Huang 243true in vitro.

METHODS: To treate them with different concentrations of tranilast 0(control group), 12.5, 25, 50, 100mg/L after culturing human retinal pigment epithelial cells(hRPE-19 cell line)3-5 generations. And then to observe cell morphology under inverted microscope combined with HE staining, identify the cell by keratin CK18. Detecting inhibition rate by MTT colorimetric method after the cells were treated with different concentrations of tranilast for 12h, 24h, 48h. The RPE cells were cultured with different concentrations of tranilast for 24h, then to measure the transforming growth factor β1(TGF-β₁)and platelet-derived growth factor receptor A protein(PDGFR-A)expressing by Western-Blot and immunohistochemical method.

RESULTS: The cell inhibition rate increased with the increase of concentration at the same time point(P<0.05). It shows a statistically significant difference. TGF-β₁ protein expression is in cell plasma and PDGFR-A protein in the cell membrane and cytoplasm, the expression of their amount was lowered with the increase of concentration of tranilast(P<0.05). It shows a statistically significant difference.

CONCLUSION: Tranilast could inhibit RPE proliferation, and it may be connected with the decreased TGF-β₁ and PDGFR-A protein expression.]]> 2013/9/23 14:08:04 Jing Yu and Dong-Bo Pang 242true METHODS: In vitro rust hRPE cells proliferation differentiation model was established. Blank control group, rust group, PDGFR-ɑ antibody(1, 10, 50, and 100μg/mL)treatment group were established. The effect of anti-PDGFR-ɑ antibody on the growth of RPE was determined by MTT colorimetric assay after 0, 12, 24, 48h,and the inhibition rates were calculated accordingly.

RESULTS: After adding the anti-PDGFR-ɑ antibody, hRPE cells proliferation activity was decreased at certain concentration as the antibody concentration increased, and 50μg/mL anti-PDGFR-ɑ antibody was the optimum concentration to inhibit hRPE cells proliferation, the inhibitory rate of hRPE was 42.44%.

CONCLUSION: The anti-PDGFR-ɑ antibody shows strong inhibitory effect on the growth of cultured hRPE.]]> 2013/9/23 14:08:04 Lin-Lin Li and Dong-Bo Pang 241true METHODS: Some healthy SD rats were operated by means of single intraperitoneal injection of 1% streptozotocin based on the standard of 50mg/kg wight, after that the blood sugar value was greater than 16.7mmol/L as DM model, then randomly divided into 3 groups, each group was 10 rats. In addition to take 10 healthy SD rats as control group. Four groups of rats were bilaterally eyeball intravitreal injection in turn with NgR-siRNA virus 10μL(siRNA group), NgR-siRNA virus diluted 10μL(DM group), NgR-siRNA virus-negative-control solution 10μL(siRNA blank group), NgR-siRNA virus diluted 10μL(normal control group), and fed normally. During that time, some life indexes like blood glucose, body mass, etc. were measured and recorded. After 12wk, the expression of NgR and Rhoa, HE staining, and TUNNEL staining were detected by Western blot analysis.

RESULTS: Western blot analysis: compared with normal control group, the expression of NgR and Rhoa in DM group and siRNA blank group increased significantly(P<0.01), while siRNA group was no significant change(P>0.05); compared with DM group and siRNA blank group, the expression of those proteins significantly lowered in siRNA group. HE staining: compared with normal control group, some extent ganglion cells arranged disorder, irregular shape, spacing not consistent were all found in three groups of model rats; compared with DM group and siRNA blank group, there was some improvement in siRNA group of ganglion cells about the order and shape size. TUNEL staining: compared with normal control group, there were retinal ganglion cells apoptosis in all of three groups of model rats. Compared with DM group and siRNA blank group, the number of retinal ganglion cells apoptotic cells was less, and the shape of cells had improved significantly in siRNA group.

CONCLUSION: In the DM phase, the expression of NgR and Rhoa were up-regulation, the condition of diabetic retinal ganglion cell apoptosis was improved after that the NgR-Rhoa-Rock signal pathways had been inhibited.]]> 2014/8/19 14:40:38 Yun-Jie Fu and Xue-Zheng Liu 240true METHODS: For 43 healthy rabbits, 40 were randomly selected for establishing CNV model in corneal stroma. The right eyes(group A)were received no medicine and the left eyes(group B)were injected dexamethasone after successfully establishing the model. The no modeling 3 rabbits were normal control group. The morphologic change of corneal was observed with slit lamp microscope and the areas of CNV was calculated every day, then 8 rabbits were randomly chosen for sacrificing at 1, 4, 7, 14, 21d respectively. The pathological characteristics of CNV were observed after HE staining, and IL-1β and TNF-α expression was detected by immunohistochemistry.

RESULTS: CNV was grown at the 4d after suture, and the 7-14d was vigorous growth period. inflammatory cell infiltration appeared after HE staining, and CNV was located at the superficial stroma of cornea. Immunohistochemistry results showed that IL-1β and TNF-α expression was gradually increased with prolonged suture time. Compared with corneal stitch group, the rabbits cured by dexamethasone were found with less inflammatory cells infiltrating and neovescularization, moreover, the expression of IL-1β and TNF-α decreased. There were statistical significance between the two groups(P<0.05).

CONCLUSION: Dexamethasone can inhibit the CNV growth by controlling the inflammation of corneal and restraining IL-1β and TNF-α expression.]]> 2014/8/19 14:40:38 Rui Shi,Yu-Shun Xue,Le Yang,Ji-Min Wang,Feng Wang and Yi-Ning Shi 239true in vitro under high glucose conditions, furthermore, to explore the effects of high glucose on epithelial-mesenchymal transition(EMT)in RPE cells.

METHODS: Human RPE(hRPE)cells were cultured in vitro. Containing a final concentration of 60mmol/L glucose was used for high glucose treatment. The cells were divided into normal glucose group(5.5mmol/L, NG)and high glucose group(24, 48 and 72h)respectively. The expression of N-cadherin and fibronectin in hRPE cells were evaluated by immunofluorescence and real-time PCR.

RESULTS:RPE cells became disorganized and swollen over time under high glucose conditions, especially in 72h subgroup. Immunohistochemical analysis revealed that the expression of N-cadherin in RPE cells under high glucose conditions was decreased compared with that in the control group, while the expression of fibronectin was increased. Real-time PCR results showed that the expression of N-cadherin mRNA in high glucose group was decreased at 24h compared with that in the control group, and declined markedly at 72h(F=12.252, P=0.000). There were no significant differences between the control group and the high glucose group at 24h, while the differences between the control group and the high glucose group(48 and 72h)were significant respectively(P<0.05). Meanwhile, the expression of fibronectin mRNA in RPE cells was increased in high glucose group at 24h, and reached the peak at 72h(F=50.543, P=0.000). There were no significant differences between the control group and the high glucose group at 24h. Compared with the control group, the expression of fibronectin mRNA in hRPE cells was increased significantly in high glucose group(48 and 72h)respectively(P=0.000, P=0.000).

CONCLUSION: The expression of epithelium marker N-cadherin is down-regulated under high glucose conditions in hRPE cells in vitro. Meanwhile, the expression of mesenchymal maker fibronectin is induced and appeared to EMT changes. Results of this study will enrich our growing understanding in proliferative diabetic retinopathy and hopefully lead to novel insights for the pathogenesis and therapeutic treatments.]]> 2014/8/19 14:40:38 Wen-Jiao Bi,Rui-Shu Li,Ding-Shan Hou,Yan Fan and Xiao-Mei Zhang 238true METHODS:After the crush models of acute optic nerve were set up, 72 males SD rats were divided into 4 groups randomly as common group without surgery, FBS negative control group, Daidzein treatment group(10μmol/L, 100μmol/L, 1000μmol/L)and positive control group using rats nerve growth factor(mNGF, 100ng/mL). Three days after interference, all experimental animals were executed. HE staining was used to evaluate morphologic change of the retina, immunohisochemical staining and western-blot tests for identifying and quantifying the distinct expression of Caspase-3 and GAP-43 among the groups.

RESULTS:Compared with the normal group and negative control group, retinal morphology of different concentrations of each Daidzein treatment group and positive control group was more complete, the expression of Caspase-3 protein was relatively lower, the expression of GAP-43 protein was relatively higher, the differences have statistically significance(P<0.05).

CONCLUSION: Daizein injection in the vitreous cavity has the capacity of protection and restoration in rat's acute nerve damages.]]> 2014/7/22 8:27:34 Zhong-Jun Tang,Zhen-Ping Huang,Wen-Jing Yang,Yong-Xiang Zou and Ji-Ping Cai 237true METHODS:hRPE cells were cultured and divided into four groups: normal glucose group(NG)(5.6mmol/L), high glucose group(HG1:15mmol/L D-glucose,HG2:20mmol/L D-glucose,HG3:30mmol/L D-glucose), PD98059 group: hRPE cells were treated by an efficient and selective inhibitor PD98059(20μmol/L)of p42/p44MAPK signal transduction pathway and solvent dimethyl sulfoxide group(DMSO group). The expression of VEGF and pigment epithelium derived factor(PEDF)mRNA was detected by RT-PCR. VEGF protein expression in cultured hRPE supernatants was detected by enzyme-linked immumosorbent assay(ELISA).

RUSULTS:VEGF mRNA and protein expression induced by elevated glucose concentration increased significantly. VEGF mRNA and protein expression were restrained in PD98059 group. Ratio of(VEGF/β-actine)/(PEDF/β-actine)in PD98059 group decreased significantly compare with that in high glucose group.

CONCLUSION:p42/p44MAPK signal transduction pathway might play a part in VEGF expression induced by elevated glucose concentration in cultured hRPE cells.]]> 2014/7/22 8:27:34 Chun-Xia Zhang,Jian-Min Hu and Ming-Zhong Zhuang 236true METHODS:To establish recombinant adenovirus with the ELOVL4 protein and green fluorescent protein, transferred into cultured PC12 cells. The cells were divided into 3 groups: PC12, PC12+Ad-GFP and PC12+Ad- ELOVL4, former two groups serve as controls. ELOVL4 gene expression was quantified by qRT-PCRs. ELOVL4 protein was analyzed by Western-Blot(WB). The transduced cells were treated with both EPA and AA(1:1). After 48h of incubation, cells were collected, total lipids extracted and fatty acid methyl esters prepared and analyzed by gas chromatography-mass spectrometry(GC-MS).

RESULTS: When supplemented together, 20:5n3(EPA)and 20:4n6(AA)were efficiently taken up at almost the same amounts in the PC12 cells regardless of ELOVL4 expression. The ELOVL4-expressing cells elongated both EPA and AA to a series of n3 and n6 VLC-PUFAs. From 20:5n3/EPA, 34:5n3 and 36:5n3 account for 0.71% and 1.6%, respectively. From 20:4n6/DHA, 34:4n6 and 36:4n6 were only 0.46% and 0.61%, respectively. The total relative mol% of n3 VLC-PUFAs synthesized from EPA was almost two times that of n6 VLC-PUFAs synthesized from AA.

CONCLUSION: ELOVL4 protein preferentially elongates n3 PUFA to VLC-PUFAs over n6 PUFA. Dietary supplementation of appropriate n3/n6 PUFAs may provide STGD3 patients with some therapeutic benefits.]]> 2014/7/22 8:27:35 Man Yu,Wei Lin,Bo Chen and Zheng-Zheng Wu 235true METHODS: CNV models were established by implantation pellets containing basic fibroblast growth factor(bFGF)into corneal stroma. CNV models were measured by biomicroscopy photography. Immunohistochemical staining and imaging analysis system were used to detect the expression of Slit3 and Robo4 in the models after 1, 4, 7, 10 and 14d.

RESULTS: The area of CNV and the expression of Slit3, Robo4 were increased in CNV models compared to that in normal cornea and reached highest level on 7d. And the expression level of Slit3 and Robo4 were significantly correlated with the size of CNV on every time point except 1d(r=0.84-0.91, all P<0.05).

CONCLUSION: The expression of Slit3 and Robo4 may be related to the CNV development. They are potential therapeutical target for CNV.]]> 2014/6/19 9:55:00 Shi-Yi Xiao,Li Wang,Ren-Dian Chen,Jin Wu,Yue-Li Zhang and Li He 234true METHODS: Total 45 New Zealand white rabbits were divided randomly into five groups(A, B, C, D, E). A groups: 100℃(n=10), B groups: 200℃(n=10), C groups: 300℃(n=10), D groups: 400℃(n=10), and E groups: control group(n=5). All left eyes of rabbits in A,B,C,D groups were induced corneal neovascularization by constant temperature burning device. The growth of CNV was observed by slit lamp microscope and the area of CNV were recorded on 4 ^th, 7 ^th, 14^th, 30^th days postoperatively. SPSS 19.0 statistical package was used for data analysis, and the data was recorded by mean±standard deviation. Comparison by analysis of variance was made by repeated measures in the area of neovascularization at each time point in groups. Statistical tests were considered significantly when P values were less than 0.05.

RESULTS: On postoperative 4^th, 7^th, 14^th, 30^th days: no neovascularization was found after corneal thermal burn in A group, but only a few nebula left(n=2); the area of CNV were(9.16±1.45)mm²,(37.73±5.49)mm²,(62.44±7.54)mm²,(40.28±7.39)mm² in B group respectively; and(11.45±1.04)mm²,(44.51±4.64)mm²,(66.13±4.13)mm²,(43.04±2.33)mm² in C group respectively; and(13.23±0.86)mm²,(47.26±4.59)mm²,(67.57±4.56)mm²,(45.59±4.44)mm² in D group respectively, and part corneal carbide(n=4)was observed as well as corneal perforation(n=6)were found on 3d in D group. No neovascularization was found in normal control group. Comparison of the areas of CNV at each time point between groups was statistically different, P<0.05. Statistical differences were found among B, C, D groups, P<0.05.

COCLUSION: In 4 to 7d, the higher the temperature is, the more the neovascularization area of CNV are. It has no significant difference in 14 to 30d. But corneal carbide and corneal perforation are often found in 400℃ group, so its modeling failure rate is high. It is between 200℃ and 300℃ that repeatability and uniformity of the corneal neovascularization model of rabbit are superior.]]> 2014/6/19 9:55:00 Yong Jia,Hua Jiang and Yong-Qiang Wang 233true in vivo.

METHODS:Three IκBα specific double stranded siRNAs were designed and synthesized. They were transfected into rat A7r5 cells which express IκBα gene. Flow cytometry was used to assess transfected efficiency. The mRNA and protein levels of IκBα were examined by Real Time quantitative polymerase chain reaction(Real Time-PCR)and western blot to screen a candidate valid sequence with the highest inhibitory rate. The Cy3 labeled non-specific control siRNA or the candidate valid siRNA was then injected into rat anterior chamber. Distribution of Cy3-siRNA in rat ciliary muscles was viewed by fluorescence microscopy, and the inhibitory effect in vivo of the valid siRNA was identified via Real Time-PCR and immunofluorescence.

RESULTS: The suppression effect of the siRNA targeting the CTACGATGACTGTGTGTTT of IκBα gene was most obvious by vitro screening. By anterior chamber injection, this valid siRNA could reach rat ciliary muscles and effectively suppress IκBα gene expression with the highest inhibitory rate of 59.0% on mRNA level at 24h after RNAi, and 52.3% on protein level at 72h after RNAi(P<0.01).

CONCLUSION: It proves that the siRNA targeting the CTACGATGACTGTGTGTTT of IκBα gene is the valid sequence to suppress rat IκBα expression of ciliary muscles by RNAi in vivo.]]> 2014/5/22 9:13:00 Rui Zeng,Yu-Qing Lan,Hai-Jun Gong,Chi Zhang and Jin-Miao Li 232true METHODS: SD rats were randomly divided into four groups: normal control group(NC), normal fluctuation group(NF), diabetes group(DM)and diabetes fluctuation group(DF). Diabetic models were established through intraperitoneal injection of STZ. A certain amount of glucose was injected in the rats of group NF and DF in an intraperitoneal mode three times a day after the model was established, thereby causing blood glucose fluctuations. Rats were killed and the retinal tissues were taken in the 8th week. Single cell gel electrophoresis(SCGE)technique was adopted for detecting DNA injury extent in the retina tissue.

RESULTS:Groups NF and DF showed significant and regular fluctuations. The curve of blood glucose fluctuations was relatively stable. All values of MBG, SDBG, LAGE and M were significantly increased compared with group NC. Group DF was increased more significantly. It was statistically significant(P<0.01). SCGE showed that there were DNA damages in different levels in the cells of group NF, DM and DF. Indicators of cells such as TL, TDNA%, TM, OTM were higher than that in group NC. It was statistically significant(P<0.01). The comparison difference between two groups was also significant(P<0.01).

CONCLUSION: Rat retinal tissues have DNA injury during early diabetic period. DNA injury is gradually aggravated with blood glucose fluctuation. It indicates that high blood glucose and blood glucose fluctuation are involved in the mechanism of cell DNA injury, and they may be one of DR early event, have played a certain role in the incidence of DR.]]> 2014/5/22 9:13:00 Chun-Liu Gai,Jing-Ru Zhao and Xiao-Long Chen 231true METHODS: Using 532nm laser photocoagulation to establish a mouse model of CNV. We observed the formation of CNV by histopathological examination after 2wk later. Forty successful models of mice were randomly divided into control group(group 1, 10 rats), normal saline group(group 2, 10 rats), endostatin group(group 3, 10 rats)and avastin group(group 4, 10 rats). The drugs were injected into the mice' vitreous after photocoagulation 2wk later. Then 1wk later, we took the mice eyeballs to perform the HE and immunohistochemical staining to observe. The statistical analysis of ANOVA was done by SPSS 16.0 and the LSD-t test was used for multiple samples, taking P<0.05 as the test standards.

RESULTS: Two weeks later, HE histopathological examination was done, light microscope showed large amount of new vessels' formation, the positive rate for CNV was 72.8%. The blank control group compared with the normal saline group P>0.05, had no inhibitory effect on CNV; endostatin treated group compared with control group, P<0.05, had a certain inhibitory effect; avastin group compared with the control group, P<0.05, had an inhibitory effect on CNV; the LSD-t was performed on Avastin group and endostatin group, P<0.05, which were statistically significant. We thought that the two drugs have different inhibitory effect on mice' CNV, because (-overx)_Avastin =26.90,(-overx)_endostatin=29.13,(-overx)_Avastin<(-overx)_endostatin, we can infer that endostar had lower inhibitory effect on mice CNV than Avastin.

CONCLUSION: Laser-induced CNV animal models of colored mice C57BL/6J is of short time and high rate establishment and it is an ideal model for CNV study. Endostar has certain inhibitory effect on CNV, and it is likely to become one of the important drugs for CNV-related diseases in the future.]]> 2014/5/22 9:13:00 Jing Li,Yong Ma,Hong-Mei Wang,An-Ming Xie and Xuan Liu 230true METHODS: Immunohistochemistical staining method(PV)was adopted to detect the expression of TNF-α and receptor I in pterygium(72 eyes)and para-pterygium conjunctival tissue(30 eyes). The relationship between the expression and clinical-pathological parameters was also analyzed.

RESULTS: The positive rates of TNF-α were 65.3%(47/72), 26.7%(8/30)in pterygium and para-pterygium conjunctival tissue. The positive expression of TNF-α had statistic difference between the two groups(χ²=12.706, P<0.01). The positive rates of TNF-α receptor I were 56.9%(41/72), 16.7%(5/30)in pterygium and para-pterygium conjunctival tissue. The positive expression of P55 had statistic difference between the two groups(χ²=13.875, P<0.01). The positive rate of TNF-α in recurrent pterygium group was higher than primary pterygium group(χ²=6.547, P=0.011). There had no statistically significance of the expression intensity between the two groups(F=1.288, P=0.393); the positive rate in advanced pterygium group was higher than quiescent pterygium group(χ²=4.082, P=0.043). The expression intensity had no statistically significance between the two groups(F=0.489, P=0.708). The positive rate of P55 in recurrent pterygium group was higher than primary pterygium group(χ²=9.907, P=0.002). There had no statistically significance of the two group's expression intensity(F=1.175, P=0.424); the positive rate in advanced pterygium group was higher than in quiescent pterygium group(χ²=11.140, P=0.001). The expression intensity had no statistically significance between the two groups(F=0.665, P=0.621).

CONCLUSION:The expression of TNF-α and P55 are changing according to the development of clinical staging and onset. The expression of TNF-α and P55 may be related to clinical classification, staging and patient's working conditions of pterygium. There has no significant difference expression intensity of TNF-α and P55 in clinical staging and onset of pterygium.]]> 2014/5/22 9:13:00 Bing Wu,Jian Yang,Jin Wei and Ping Ma 229true METHODS: Fifty-four mice were randomly divided into two groups which were fed a high-fat diet for 19wk. One group mice were lavaged LOM by the dosage of 80mg/kg daily at the same time. The obese mice were selected and divided into diet-induced obesity(DIO)group, diet-induced obesity and lomerizine(DIO+LOM)group. The mice in the control(CON)group were fed a basal diet. The ultrastructural changes of RGCs were detected by transmission electron microscope. The cellular apoptosis was detected by TUNEL. The laser scanning confocal microscope was used to measure intracellular calcium ion concentration.

RESULTS: Compared with the CON group, the RGCs in DIO group showed smaller and condensation of nuclear chromatin and increased electron density of the cytoplasm, whereas the changes in DIO+LOM mice were obviously diminished. TUNEL staining showed that the number of apoptosis cells in the ganglion cell layer(GCL)increased in DIO group and the percentage of apoptotic cells was much higher than that in the CON groups(P<0.01). The DIO+LOM group mice showed fewer apoptosis cells and the percentage of apoptotic cells in this group were significantly decreased than the DIO mice(P<0.05). The Laser scanning confocal microscope detection showed Ca²⁺ staining intensity of RGCs in DIO group increased and its staining intensity ratio was significantly higher than in CON group(P<0.01), the Ca²⁺ staining intensity and its staining intensity ratio in DIO+LOM group were significantly decreased than the DIO group(P<0.01).

CONCLUSION: Lomerizine has neuroprotective effects on damage of retinal ganglion cells in diet-induced obesity mice, which may be related to the attenuation of intracellular Ca²⁺ overload.]]> 2014/3/24 14:01:48 Xia Bai,Jian Zhao,Wen-Qing Zhao and Yu-Ling Chen 228true METHODS: Totally 140 Wistar rats were randomly divided into normal control group(group A)20, high intraocular pressure control group(group B)60 and high intraocular pressure progesterone treatment group(group C)60. The acute high intraocular pressure models were made by saline anterior chamber perfusion. Group C were treated with intraperitoneal injection of progesterone 4.0mg/Kg immediately after intraocular pressure restored, and after 6, 24h and every other day were treated with subcutaneous injection. Group B were injected with equivalent saline at the same time. At the 1, 3, 7d after ocular hypertension, the retinas of rats in each group were obtained respectively. Retinas pathological changes were observed in samples hemetoxylin and eosin staining. The protein expression of Caspase-3 was measured with immunohistochemistry and Western blot. Apoptosis in retinal nerve cells were tested with TUNEL technique. Statistical analysis was performed in each group.

RESULTS: The rats retina of group B and group C were edema, but the degree in group C was lower than that in group B. In relative quantitative detection of immunohistochemistry and Western blot, expression of Caspase-3 was lower than group B. Substantial expression of Caspase-3 protein lay in cell nucleus and cytoplasm. The level of Caspase-3 in group B was higher than in group C. The positive cells were mainly in inner nucleus layer and ganglion cell layer. The level on the first day after high intraocular pressure was the highest, then gradually decreased along with the passage of time. Apoptosis tested with TUNEL technique, positive cells could be observed in group B and group C, which in inner nucleus layer and ganglion cell layer. But the number in group B was significantly more than group C.

CONCLUSION: Progesterone has nerve protection functions on retinal acute high intraocular pressure injury in rats.]]> 2014/3/24 14:01:48 Ye-Hong Cao,Dian-Wen Gao and Le-Li Lin 227true METHODS: Adult male Spraque - Dawley(SD)rats were randomly divided into control(CON), diabetes mellitus(DM), lomerizine(LOM)group, and each group had 40 rats. Diabetes rat model was induced by Streptozotocin(STZ)of intraperitoneal injection of 60 mg/kg of disposable. In LOM group, after model was established, rats were lavaged LOM by the dosage of 60mg/kg daily, the CON group and DM group were given the same dosage sodium chloride. In 4^th, 8^th, 12^th wk, RGCs' apoptosis were detected by HE, TUNEL, transmission electron microscope, and TUNEL and laser confocal microscope detection was used to test the calcium ion concentration.

RESULTS: Morphological observation: with the extension of the DM, RGCs decreased gradually and appeared disordered arrangement of cells. In DM group, different stages of apoptosis were observed by transmission electron microscope and got worse gradually with its extension. In LOM group, compared with DM group in the same period, RGCs apoptosis signs diminished gradually at 8^th and 12^th wk. TUNEL detection: no apoptotic RGCs was observed in CON group. In DM group, few TUNEL positive RGCs were seen at 4^th wk, and became more and more gradually. The apoptosis index was significantly higher in DM group compared with CON group in same time and there was statistical significance(P<0.01). In LOM group at 8^th and 12^th wk, compared with DM group in the same period, the numbers of TUNEL positive RGCs decreased. The apoptosis index was significantly lower in LOM group compared with DM group in the same period(P<0.01). Calcium ion concentration detection by laser confocal microscope: compared with CON group in same time points, at 8^th and 12^th week, DM group's calcium fluorescent staining intensity of RGCs markedly elevated and had significant differences.(P<0.01). In LOM group, at 8^th and 12^th wk, calcium fluorescent staining intensity of RGCs markedly decreased and had statistical significance(P<0.01).

CONCLUSION: The LOM played a protective role for RGCs in early stage of diabetic rats.]]> 2014/3/24 14:01:48 Rui-Dong Gu,Ying-Ying Hao and Xiao-Long Chen 226true METHODS: Under different experimental conditions such as UVA exposure, hyperosmotic stress condition and hypoosmotic stress condition, RPE cells were cultured for different time periods. The betaine /γ-amino- n-butyric acid(GABA)transporter, the sodium-dependent myoinositol transporter and the taurine transporter(TAUT)mRNA were measured by quantitative PCR. The radioactive labeled osmolytes were measured to evaluate the level of osmolytes transportation.

RESULTS: This study demonstrated that RPE expressed mRNA specific for the betaine/GABA transporter, for the sodium-dependent myoinositol transporter and for the TAUT. In comparison to norm osmotic(300mosmol/L)controls, a 3-5-fold induction of mRNA expression for the betaine/GABA transporter, the sodium-dependent myoinositol transporter and the TAUT was observed within 6-24h after hyperosmotic exposure(400mosmol/L). Expression of osmolyte transporters was associated with an increased uptake of radioactive labeled osmolytes. Conversely, hypoosmotic(200mosmol/L)stimulation induced significant efflux of these osmolytes. UVA significantly stimulated osmolyte uptake. Increased osmolyte uptake was associated with upregulation of mRNA steady-state levels for osmolyte transporters in irradiated cells.

CONCLUSION: UVA induces osmolyte uptake in RPE. It is similar reaction to hyperosmotic stress. This suggests that osmolyte uptake response by UVA may be important to maintain homeostasis.]]> 2014/3/24 14:01:48 Da-Yang Wu and Jin-Song Zhang 225true in vitro.

METHODS: MSCs were isolated and attached to the wall of culture dishes by their specific adherent ability. Then the cells were characterized by flow cytometry. The neurosensory retina was isolated from retina of SD rat and it was tested by hematoxylin-eosin(HE)staining. The pathological changes of light-injured neurosensory retina was observed under transmission electron microscope. Three kinds of supernatant fluid of light-injured neurosensory retina of SD rats were prepared. The third passage of MSCs were cultured with these mixed medium for 7-8d, we used RT-PCR to see whether they could express rhodopsin, neuron-specific enolase(NSE), and glial fibrillary acidic protein(GFAP), and positive cells were counted and analyzed.

RESULTS: HE staining showed the retinal sheets included full-thickness neural retina. Neurosensory retina developed ultrastructural destructions by light injury. RT-PCR showed that the medium of mixed I expressed higher positive rate of rhodopsin(0.3915±0.00644), NSE(0.2019±0.00682), GFAP(0.1972±0.00211)than the medium of mixed Ⅱ rhodopsin(0.0983±0.00319), NSE(0.1048±0.00323), GFAP(0.1040±0.00254)and medium of mixed Ⅲ rhodopsin(0.0044±0.00126), NSE(0.0498±0.00149), GFAP(0.0467±0.00333). The difference of intergroup has statistical significance.

CONCLUTION:The supernatant fluid of light-injured neurosensory retina of SD rats can induce MSCs to differentiate into retina-like cells and provide new insights of stem cell therapy for retinopathy.]]> 2014/2/27 9:12:34 Yue Bai and Guo-Xing Xu 224true METHODS: Mice were fed high-fat diet. After 19 weeks of feeding, the mice were divided into diet induced obesity-resistant(DIO-R)group and diet induced obesity(DIO)group, while mice of the control(CON)group were fed a basal diet at the same time. The apoptosis of RGCs was detected by TUNEL. Laser scanning confocal microscope was used to detect the intracellular calcium ion concentration.

RESULTS: TUNEL staining showed apoptosis cells in ganglion cell layer(GCL)in DIO group increased and the percentage of apoptotic cells was(6.7±1.2)% which was much higher than in CON and DIO-R groups(P<0.01, P<0.05). There was no significant difference between CON group and DIO-R group(P>0.05). Laser scanning confocal microscope detection showed Ca²⁺ staining intensity of RGCs in DIO group increased and its staining intensity was significantly higher than in CON and DIO-R mice(P<0.01,P<0.01), whereas there was no significant difference between CON group and DIO-R group(P>0.05).

CONCLUSION: Intracellular calcium ion overload might be involved in the RGCs apoptosis in the diet-induced obese C57BL/6 mice.]]> 2014/2/27 9:12:34 Xia Bai,Jian Zhao,Wen-Qing Zhao and Yu-Ling Chen 223true METHODS: Sixty SD rats were randomly divided into two groups: control group(CON)and diabetes mellitus group(DM). Diabetic rat model was produced by intraperitoneal injection of 1% STZ in 30 adult male SD rats. At 4, 8, 12wk,the rats were killed and eyeballs were enucleated for the HE staining, TUNEL staining, transmission electron microscopy detection respectively, and laser confocal microscope detection was used to detect the calcium ion concentration.

RESULTS:At 8wk RGCs decreased gradually and appeared disordered arrangement and got worse at 12wk in DM group. In DM group, mitochondrial swelling was detected at 4wk., and became more obvious, more in number at 8wk with reduction in some cells' volume and the number of organelles decreased. In DM group, few TUNEL positive RGCs were seen at 4wk, and became more and more at 8 and 12wk. The apoptosis index was significantly higher in DM group compared with CON group in different time points(P<0.01). Intracellular calcium ion concentration of RGCs in DM group began to elevate obviously at 8 and 12wk in contrast with CON group in the same time point(P<0.01). In DM group, Intracellular calcium ion concentration of RGCs increased difference was statistically significant between 8 and 12wk(P<0.05).

CONCLUSION: The study suggested that RGCs apoptosis occurs in early stage of diabetes, the mechanism might be associated with increased intracellular calcium ion concentration.]]> 2014/2/27 9:12:34 Rui-Dong Gu,Ying-Ying Hao and Xiao-Long Chen 222true METHODS: Eighty 7-day-old SD rats were randomly divided into normol group, high oxygen group, 17β-estradiol treatment group and vegetable oil treatment group. Rats of the high oxygen group were put into the environment exposed to 75% oxygen for 5d and backed to room air for another 5d to establish the oxygen-induced retinopathy model. The treatment group was given 17β-estradiol by injection with the dose of 0.5uL/rat/d(2μg/μL)before exposed to 75% oxygen. The vegetable oil treatment group is similar to 17β-estradiol treatment group but the medicine changed to vegetable oil. Counting the endotheliocyte nuclei of new vessels which extended from retina to vitreous body in the tissue-slice of HE staining, and investigate the change of retinal blood vessels by using methods of retina flat-mount. The expression of VEGF in retina by immunohistochemistry and reverse transcription-polymerase chain reaction(RT-PCR)analysis.

RESULTS: Rats treated with E₂ showed less neovascularization than high oxygen group and oxygen-exposed rats treated with vegetable oil(P<0.05). Adenosine diphosphate-ase(ADP ase)stained retina flat- mount showed that: less free-vascular areas and new blood vessels were seen in the 17β-estradiol treatment group compared with the high oxygen group. The expression of VEGF in 17β-estradiol treatment group lower than high oxygen group and vegetable oil treatment group, but higher than normal group in immunohistochemistry and RT-PCR analysis.

CONCLUSION:17β-estradiol has prevention effects in retinal neovascularization and the mechanism may involve in its interaction with VEGF.]]> 2014/2/27 9:12:34 Hai-Yan Fan,Wei- Jie Ni,Cai- Hong Shi,Jian Jiang,Rong Huang,Jun-Fang Wang,Hai-Lin Hu and Zhong Chen 221true METHODS: OCM-1 cells were cultured under normoxia and hypoxia in vitro. We added chemical hypoxia inducer cobalt chloride(CoCl₂)into nutrient medium at the concentration of 100μmol/L to simulate hypoxia microenvironment inside tumor to culture cells of hypoxic group. And hypoxic group cells were divided into five groups: simple hypoxic group, interference group, positive control group, negative control group and liposome group. SiRNA(including HIF-1α-siRNA, β-actin-siRNA and negative control)were designed and synthetized in vitro. Lipofectamine^TM 2000 was taken to transfect siRNA into OCM-1 cells under hypoxia. RT-PCR and Western blot were used to check the expression status of HIF-1α and vimentin on mRNA and protein levels before and after hypoxia culture and cell transfection.

RESULTS: Compared with normoxia group, there was no obvious change on the expression level of mRNA of HIF-1α in simple hypoxia group(P>0.05), while the expression level of its protein increased obviously(P<0.01); both mRNA and protein levels of vimentin were up-regulated(P<0.01). Compared with other hypoxic groups, the expression level of mRNA of β-actin was down-regulated in positive control group(P<0.01), which indicated that our operation of cell transfection was successful. In interference group, the expression of HIF-1α and vimentin were down-regulated obviously both on mRNA and protein levels(P<0.01). There was no significant difference in the expression of HIF-1α and vimentin in negative control group and liposome group(P>0.05).

CONCLUSION: Hypoxia can up-regulate the expression of HIF-1α on protein level in OCM-1 cells, and activate the transcription of vimentin as the downstream gene of HIF-1α, up-regulate the expression of vimentin both on mRNA and protein levels. This hints that HIF-1α can regulate the EMT in uveal melanoma and plays an important role in the process of tumor invasion and metastasis. We successfully down-regulate the expression of HIF-1α and vimentin by transfecting OCM-1 with HIF-1α-siRNA. This suggests that suppression the expression of HIF-1α at the molecular level maybe can shut down the process of tumor invasion and metastasis and offer new directions for cancer treatment.]]> 2014/2/27 9:12:34 Juan Zhao,Zhan-Yu Zhou,Shuang Wang,Fu-Xiang Yuan and Fa-Yan Lu 220true METHODS: Studies were performed with control group, high glucose concentration(HGC)group, SiNgR group(HGC medium including AAV2-SiNgR virus)and SiRNA control group(HGC medium including the negative sequence of nucleotides). Three days after culture, RGCs was observed by microscope, expressions of NgR, Rock and F-actin were detected by Western blot, cell vitality was determined by MTT, and cell morphology was also detected by F-actin immunohistochemical staining.

RESULTS: Compared with control group, in HGC and control SiRNA groups, cell volume decreased with less processes, light refraction was strengthened, and the expressions of Rock and NgR were significantly increased(P<0.05, respectively), while the expression of F-actin was reduced(P<0.05); Compared with control group, there was no obvious difference in the expression of NgR, Rock and F-actin, and cell vitality between SiNgR group and control group(P>0.05).

CONCLUSION: The activation of NgR-Rock signaling pathway may plays an important role in HGC impairing RGCs.]]> 2014/1/20 9:29:53 Yu-Zhi Han,Xue-Zheng Liu and Zhong-Fu Zuo 219true METHODS: Fresh porcine corneas were drilled down by 13mm-diameter trephine. Then the samples were put into four kinds of cryoprotectants in turn for cooling equilibrium with “simplified four-step” method and then stored in liquid nitrogen. All samples were thawed in water at 40℃ before being used. The samples treated with dexamethasone were made and the preceding three cooling steps were same. But the samples were additionally incubated in 0.01, 0.03, 0.05mg/mL dexamethasone for 18h at 4℃ before the last cooling step. Totally 50 BALB/c mice were randomly divided into 5 groups including 10 mice in each group: the control group; routine cooling group which were treated through four cooling steps; dexamethasone group I, dexamethasone group II and dexamethasone group III which were treated with 0.01, 0.03, 0.05mg/mL dexamethasone successively. After treatment, corneal samples were transplanted into the hypodermis on the back of mice. After 14d, the samples were taken out, imbedded in paraffin, and stained by HE and CD25/FasL immunohistochemistry. All the samples were investigated under light microscope.

RESULTS: On the 14th day, all the samples were taken out. We found the samples which adhered to surrouding tissues swelled obviously and showed translucent and a bit yellow. Through HE staining, fresh corneal samples in the control group were infiltrated with large amount of lymphocytes throughout each layer; the amount of infiltrating cells in the routine cooling group was less than the control group in which most cells were distributed in endodermis and epitheliums. In the dexamethasone-treated groups, the amount of infiltrated cells were the least. Immunohistochemical results showed that compared with the the routine cooling group and dexamethasone-treated groups, the amount of CD25 and FasL in the control group was the most, the difference was significant. Compared with dexamethasone-treated groups, the amount of CD25 and FasL positive infiltrating cells in the routine cooling group were more. ALL the differences were significant. But there was no significant difference in the amount of CD25 and FasL positive infiltrating cells among each dexamethasone treated group.

CONCLUSION: Corneas in dexamethasone-treated groups showed the lowest immunogenicity. Corneal immunogenicity was irrelevant with the concentration of dexamethasone at the range of 0.01-0.05mg/mL.]]> 2014/1/20 9:29:54 Fang Liu,Guo-Ping Xiong,Jing Wang,Li Wang,Chao Peng and Yan-Ran Huang 218true METHODS:The Tenon's capsule tissue of fresh eyes(<6h)of our hospital eye bank was taken, in vitro culture of fibroblasts was done by tissue block culture; A flow cytometry(FCM)was used to evaluate the effect of different concentrations of HCPT(0, 0.25, 1, 4mg/L)and mitomycin C(MMC: 0, 0.025, 0.1, 0.4mg/L)on HTFs cell cycle; Trypan blue staining was adopted to determine whether the inhibitory effect of HCPT and MMC on HTFs was caused by their cytotoxicity; RT- PCR was employed to detect the level of Smad7mRNA gene expression of HTFs after HCPT and MMC were used for 24h.

RESULTS:HTFs were cultured successfully in vitro, and can be used in our study; compared with the control groups, the cell cycle of HTFs which was affected by different concentrations of HCPT was significantly different in G2 phase(P<0.05). As for MMC, there were significant differences in G1 phase between groups with different concentrations of MMC and the control group(P<0.05). There was no significant difference in the rate of HTFs living cells between HCPT group(or MMC group)to which HCPT and MMC were added for 24h and the control group(P>0.05). After HFTs was affected by 0.4mg/L MMC or 4mg/L HCPT for 24h, RT PCR found that the level of Smad7 mRNA expression was significantly increased(P<0.05).

CONCLUSION: HCPT mainly blocks HTFs in G2 phase, MMC mainly impacts the G1 phase. The inhibitory effect of HCPT and MMC on HTFs proliferation is not relevant to cytotoxicity induced by the two drugs. The mechanism that HCPT inhibits the proliferation of HTFs may be due to the increasing Smad7 mRNA expression to block TGF-β signaling pathway.]]> 2014/1/20 9:29:54 Sha-Sha Luo,Lian Fan,Song Sun and Zhi-Feng Wu 217true + T cells, and to study its inhibitory effect on the thyroid-associated ophthalmopathy(TAO)in mice.

METHODS: The shRNA sequence with good disturbing potency towards AHNAK1 was designed and selected; then the shRNA was packed into lentivirus; and the CD4⁺ T cells were infected. The infected CD4⁺ T cells of mice by the packed lentivirus were observed to detect the inhibition effect on T cells. And then the immunotherapeutic effects of AHNAK1^-/- on TAO were observed by experimental animal model.

RESULTS: The shRNA with good disturbing potency was successfully screened and correctly inserted into the lentivirus. The titer of the recombinant lentivirus was 1.0×10⁶TU/mL. The CD4⁺ T cells infected by lentivirus showed the anergy trend and restrained the immune response of inflammation. Suppressing the expression of AHNAK1 in T cells of animal model can effectively control the occurring and proceeding of TAO, which can significantly reduce the expression of IL-2 /IL-1β/IFN-γ in the T cells of the control group.

CONCLUSION: This paper successfully constructs mice model with the recombinant lentivirus expressing AHNAK1 shRNA, which has a favorable inhibitory effect on secretion of IL-2, IL-1β, IFN-γ by T cells. The recombinant lentivirus can effectively inhibit the occurring and proceeding of TAO in mice.]]> 2014/1/20 9:29:54 Hua-Xin Chen,Shao-Shuo Yu,Wei-Ming Zhou and Yi Zhang 216true METHODS: Specific shRNA expression vector was constructed according to the design principles of TGFβ-R2 mRNA human GeneBank siRNA synthesis and transfected into cultured 293T with TGFβ-R2 over expression vector. After effective RNAi vector was selected by Western blot, then amplification, purification, and titration. lentivirus-TGF beta-R2 shRNA was transfected into LECs, then TGFβ-R2 mRNA expression in these cells was detected by the real-time fluorescence quantitative PCR detecting system(qPCR).

RESULTS: TGFβ-R2/GV115RNAi#1 target gene expression knockdown effect is the most obvious; the titer of packaging lentivirus is 8E+8 TU/mL; the knockdown effect is significant, the interfering efficiency is 78.1%(P<0.05).

CONCLUSION: TGFβ-R2 RNAi vector was successfully constructed, and can inhibit the expression of TGFβ-R2 mRNA in human LECs.]]> 2013/12/23 10:55:36 Zhen Li,Qing Xu and Hui Zhang 215true METHODS: A half of 20 pairs of rabbit corneas just were cultured in ACF medium with a supplement of 10% HES 130/0.4 for 28d as the experimental group. The control was cultured in ACF for 28d and then dehydrated in 5% dextran T500 for 48h. The evaluation parameters included the endothelial cells viability, the mean corneal central thickness before and after preservation, the mean water content after storage, the corneal transparency and folding, F-actin expression of corneal endothelium using Western blotting, and the electron microscopy observation of corneal endothelial cells.

RESULTS: After storage, the mean endothelial cell density of the HES 130/0.4 experimental group was 2 371±159/mm². The experimental group also exhibited a thinner corneal thickness, better transparency and less folding compared with the control. After additional dehydration, the control became thin and transparent, however, its final endothelial cell density decreased greatly to 2 138±182/mm². F-actin could be seen in corneal endothelium of two groups using Western blotting, whose expression level achieved higher in the experiment group. The ultrastructure of endothelial cells of the experiment group remained good compared with normal cornea.

CONCLUSION: HES 130/0.4 has low toxicity, and is well tolerated for endothelial cells and can be used as a continuous supplement during organ culture. It also avoids excessive cornea swelling, simplifies the storage steps, reduces infection risk, and appears to be an alternative deswelling additive in cornea organ-culture.]]> 2013/12/23 10:55:36 Jie Wei and Hua Jiang 214true METHODS: Mouse of specific Smad4 conditional knockout in lens ectoderm(Le-Cre; Samd4fl/fl or also called mutant mouse)was obtained by mating the Pax6 promoter-driven Cre transgenic mouse(Le-Cre)with Smad4 wildtype mouse(Smad4 fl/fl). To confirm that Smad4 has been conditionally inactivated only in the specific tissue of ectoderm such as lens, cornea and ectoderm of the eyelids so on. A series of assays were carried out to reveal the validity and specificity of Smad4 gene knockout at molecular and cellular levels, including genotyping by PCR, detection of green fluorescence protein(GFP)in specific tissue and Smad4 protein. The expression of Le-Cre from Lacz staining using ROSA26 reporter genes in specific ocular tissue of mice can be visualized. Preliminary phenotype of mutant mouse was also observed.

RESULTS: As early as around E10.0, strong GFP expression was observed in the embryonic lens and periorbital ectoderm of the mice, which showed Le-Cre was expressed in specific target tissue. Through genotyping for Smad4, Cre and Rosa genes, the mice were determined if they have carried Cre, Smad4 allele or Rosa reporter gene. It was further confirmed by lens-sampled genotyping that Smad4 gene was removed from some specific tissue such as lens. The spatial-temporal expression and tissue specificity of Le-Cre recombinase was also revealed by LacZ staining of Rosa; Le-Cre double transgenic mouse. According to Immunohistochemical staining, Smad4 was widely expressed in normal embryonic eyes, mainly appearing in the cytoplasm at the early embryonic stage and were transferred to nucleus with gestation developing, while in mutant embryonic eyes, Smad4 was void of expression in Cre-expressed tissues. It was observed that Smad4 mutant mouse could survive the conditional gene knockout. But those mice showed abnormal appearance such as microphthalmia, sunken socket, abnormal eyelid opening, and eyelid closure failure and periocular hair loss.

CONCLUSION: In this study, mouse model of Smad 4 conditional knockout is precisely established and lack of expression of Smad4 in mutant mice is confirmed by related genetic and proteic detection. Lack of Smad4 expression in specific ocular tissues of mutant mouse can result in the abnormality of eye and adnexa, which provides a reliable animal model to investigate the ocular development and the roles of Smad4 on it.]]> 2013/12/23 10:55:36 Ying Liu and Ding Lin 213true METHODS: A total of 24 male Brown-Norway rats were selected including 18 rats as CNV model and 6 as normal comparison. A total of 18 model rats were divided randomly into control, naringenin and its complexes of copper group with 6 rats in each group. Naringenin and its complexes \〖20mg/(kg·d)\〗 were given once-daily through intraperitoneal(i.p.)injection after laser treatment for 30d in two experimental groups, respectively. Complex samples of retina, choroid membrane and sclera in each group were taken after 30d of the establishment of animal model. The sizes of CNV lesions were observed under fluorescence microscope. Samples of choroid in each group were taken. The expression of protein and mRNA of COX-2, VEGF, PI3K, MAPK, MMP-2 and MMP-9 were examined by Western blotting and RT-PCR method at 30d.

RESULTS: The animals treated with naringenin and its complexes of copper showed significant decrease in their average CNV sizes, compared with that of the vehicle-treated animals(P<0.01), especially with naringenin's and its complexes of copper group. Compared with the vehicle-treated animals, the mRNA and protein expressions of VEGF, COX-2 and so on were significantly higher than that in the model control group(P<0.01). The expression of gene and protein in each experimental group both decreased when compared with model group. Except PI3K, MMP-9, the reduction degree in the naringenin and its complexes of copper group were larger than that in the single naringenin group(P<0.05). The differences of reduction degree were statistically significant.

CONCLUSION: These results indicate that the biological activity of naringenin can be enhanced by the formation of its complexes, and that complexes of naringenin have stronger inhibition effect on the formation of CNV in rats induced by laser.]]> 2013/12/23 10:55:36 Yan Shao,Li Hang,Hai-Tao Yu,Xue-Wen Yang,Shu-Hua Ding and Xin-Rong Xu 212true METHODS:Human lens epithelial cells SRA01/04(LECs)were treated by low concentration of glucose(5.5mmol/L). High concentration of glucose(30.5mmol/L)was used to treat the cells in order to form the high glucose model. According to adding AG490 or not, cells were divided into the control group and the experimental group, appropriate concentration 10μmol/L and 50μmol/L of AG490 were chosen and acting time of 6, 12, 24, 48h were selected. Effect of AG490 on cell migration was measured by wound healing test. The expression of TGF-β₁, FN, α-SMA mRNA were examined by RT-PCR.

RESULTS:With the prolonged acting time(6, 12, 24 and 48h), cell activity increased in the HG group, as well as more expression of TGF-β₁, FN, α-SMA mRNA were detected compared to the LG group(P<0.05). In AG490 group, the cell migration activity and expression of TGF-β₁, FN, α-SMA mRNA decreased compared to the HG group(P<0.05).

CONCLUSION:JAK-STAT pathway takes part in high glucose-induced epithelial-mesenchymal transition in human lens epithelial cells. The mechanism is that it impacts the transcriptional expression of TGF-β₁ and extracellular matrix. AG490, a JAK inhibitor, inhibits high glucose-induced epithelial-mesenchymal transition in human lens epithelial cells, And the inhibition enhances with the increasing concentration of AG490.]]> 2014/12/2 9:06:28 Li Feng,Liang Zhang,Xiao Han,Bo Lu,Xin-Ling Wang and Qi-Chang Yan 211true METHODS: The murine retinal neovascularization were induced by hyperoxia exposure.The morphological observation of retinal neovascularization was performed using angiography by fluorescein dextran injection under the fluorescence microscope, and the new blood vessels were quantified after 5d in room air(17-day-old)by counting the vascular epithelial cell nuclei protruding into viteous cavity using HE stain. Realtime PCR and Western blot were used to examine retinal Islet-1 level in postnatal 7,12, 14,17 and 26d respectively.

RESULTS: A lots of new blood vessels were demonstrated in the mouse retina in hyperoxic group by fluorescein angiography and histological method. Moreover, no significant difference was found in retinal Islet-1 level in postnatal 7d between hyperoxic group and control group, but was significantly higher in postnatal 12, 14 and 17d mice compared with control mice. However, mice at postnatal 26d, expression of Islet-1 in retina decreased to normal level.

CONCLUSION: In processing mouse model of retinal neovascularization, sustained hypoxia retinal tissue induce retinal neovascularization by increas the expression of transcription factor Islet-1.]]> 2014/12/2 9:06:28 Si-Qi Xiong,Hai-Bo Jiang,Hui-Zhuo Xu and Xiao-Bo Xia 210true METHODS: Two hundred healthy C57BL/6J mice were chosen and randomly divided into control group, hyperxia group, hyperxia control group and CCN1 treated group, with 50 mice in each group. The hyperxia control group was treated with vector plasmids by intravitreal injection. The CCN1 treated group received CCN1 siRNA recombinant plasmids by intravitreal injection. Adenosine diphosphate-ase(ADPase)stained retina flat-mounts was performed to assess the retinal vascular profiles, HE staining was applied to count the number of vascular endothelial cell nuclei breaking through the internal limiting membrane, protein and mRNA level expression of CCN1 were measured by immunohistochemistry, Western blot and RT-PCR.

RESULTS: There were large nonperfusion area and a large number of vascular endothelial cell nuclei breaking through the internal limiting membrane(25.25±1.26; 23.12±1.16)in the hyperxia group and the hyperxia control group. Regions of nonperfusion and vascular endothelial cell nuclei(8.47±1.15)were decreased in the CCN1 treated group compared to the hyperxia group and the hyperxia control group. Compared with the control group, there were high protein and mRNA expression of CCN1 in the hyperxia group and the hyperxia control group. The expression of CCN1 protein and mRNA were decreased in the CCN1 treated group compared with the hyperxia group and hyperxia control group(all P<0.05).

CONCLUSION: The abnormal expression of CCN1 has close relation with RNV. The development of RNV can be markedly inhibited by RNA interference targeting CCN1, which, we believe, will provide new molecular targets and a rationale for clinical developing new strategy for ROP therapy.]]> 2014/12/2 9:06:28 Yu Di,Yi-Ou Zhang,Yang Yang and Xiao-Long Chen 209true METHODS: Sixty eyes of 30 healthy white rabbits were operated by lens cortex removal in cataract surgery, and 30 right eyes were divided in treatment group and the other 30 eyes were divided in control group. From the first postoperative day, the control group eyes were dropped with normal saline 6 times each day, and the treatment group eyes were dropped with 1% CsA 6 times each day. Six rabbits were selected randomly and killed on the day before dropping and 1, 2wk, 1 and 2mo of postoperative day respectively. The lens of those killed rabbits were removed by surgery. The strategies of immunohistochemistry and mount in situ hybridization were used to detect the content of proliferating cell nuclear antigen(PCNA), gene of phosphate and tension homology deleted on chromsome ten(PTEN)mRNA, Ser473-R, respectively.

RESULTS: The expression of PCNA and Ser473-R were both down-regulate after operation in treatment group and control group, and the PCNA levels were significantly lower among treatment group than those in control group on 1wk(0.690±0.035 vs 0.785±0.015, t=6.099, P<0.01)and 2wk(0.571±0.038 vs 0.670±0.037, t=4.585, P<0.01). In addition, the levels of Ser473-R were significantly lower among treatment group than those in control group on 1wk(0.374±0.031 vs 0.435±0.030, t=3.486, P=0.006)and 2wk(0.220±0.022 vs 0.251±0.020, t=2.516, P=0.031). However, the expression levels of PTEN mRNA were continually increased 1wk～1mo after operation, in which the expression levels of PTEN mRNA were significantly higher among treatment group than those in control group on 1wk(0.302±0.027 vs 0.255±0.038, t=2.474, P=0.033).

CONCLUSION: 1% CsA could inhibit the proliferation of epithelial cells in lens of rabbits with after cataract through preventing PI-3k pathway.]]> 2014/12/2 9:06:28 Ning Zhao,Rui-Jun Zhang,Yi-Fan Zhong,Lei Liu and Jia Li 208true METHODS: The young SD rats(3mo)was chosen and randomly divided into 2 groups, which were experimental group and normal control group. CNV model was established by alkali burn, and the length and area of CNV was observed everyday after operation by slit lamp. After that, the expression of HIF-1α and EPO was measured by SABC and RT-PCR methods at 1, 3, 5, 7, and 14d after alkali burn. The data was analyzed by SPSS 20.0.

RESULTS: The area of CNV was increasing at 1, 3, 5, 7, and 14d after alkali burn, and the peak point appear at 7d. The growth speed was decreased after 14d. SABC method told us that no HIF-1α and very tiny amount EPO was detected at normal rats' corneal. The expression of the two factors increased at 1d after alkali burn in corneal epithelium and endoderm. The results of RT-PCR showed that a few amounts of HIF-1α and EPO mRNA were detected at normal group. The expression of the two factors was increased at 3d after alkali burn, and the peak value was found at 7d, however, it was decreased at 14d. Statistical difference was found at different time(P<0.05).

CONCLUSION: HIF-1α and EPO is closely related to CNV.]]> 2014/12/2 9:06:28 Ji-Min Wang,Rui Shi,Hui-Ling Wei,Yong Ma and Dan Gao 207true in vitro, and evaluate the effect of thioltransferase(TTase)against oxidative stress in diabetic cataract.

METHODS: Lenses were incubated with different concentrations of glucose: control group(5.5mmol/L), high glucose groups(35.5, 65.5, 95.5mmol/L)and mannitol control group for 7d. Following incubation, the lenses were evaluated daily using a dissecting microscope. After 7d of incubation, lenses were homogenized in lysis buffer and processed for measurement the activity of TTase, catalase(CAT), superoxide dismutase(SOD)and the specific value of GSSG/T-GSH.

RESULTS: The lenses treated with high glucose exhibited mistlike opacity in the early stages, while the lenses of control group remained relatively transparent. With the rise of the concentration of high glucose, the activity of TTase increased and the maximum appeared in 35.5mmol/L group(1.743±0.20mU/mg protein, P<0.01). No changes were observed in the mannitol control group(P>0.05). The specific value of GSSG/T-GSH of lenses with high glucose groups increased compared with the control groups, there were 2.89, 2.57 and 2.42 times to the normal control group(P<0.05). There was no significant difference between each group treated with high glucose, and the difference was also no significant between the mannitol group and the control group. Compared with the control group, the activities of CAT and SOD decreased in the high glucose treatment groups, the activities of SOD were 0.71, 0.52 and 0.49 times to the normal control group(P<0.05), and the activities of CAT were 0.47,0.56 and 0.50 times to the normal control group(P<0.05). There was no significantly different between each group treated with high glucose.

CONCLUSION: The antioxidant system of the lens was activated by the high glucose following the increased activity of TTase, the specific value of GSSG/T-GSH, and the decreased activities of CAT and SOD. The gradually decreased oxidation resistance in the lens may contribute to the development of diabetic cataract.]]> 2014/10/31 11:03:29 Gang Yu and Hong Yan 206true METHODS: Thirty clean pregnant Wistar rats were divided into 6 embryon groups,10-d, 12-d, 14-d, 16-d, 18-d and 20-d embryo. Two embryons were randomized obtained from every pregnant rat. One of the eyeball samples that were parallel to sagittal axis of optic nerve were cut into serial sections, used HE staining and examined by light microscope. Expression of HIF-1α protein in lens was detected by immunohistochemistry. The positive expression of HIF-1α mRNA of the other eyeball samples was detected by real-time PCR.

RESULTS:In the 10^th d of embryo(E10), the formation of lens vesicle were recognized under the light microscope. In the 12^th d of embryo(E12), the anteriorly situated cells and posteriorly situated cells have already differentiated. The anteriorly situated cells were epithelium. In the 14^th d of embryo(E14), primary fibers which came from posteriorly situated cells were examined. In the 16^th d of embryo(E16), the lens epithelium undergoes extensive proliferation, and enlongate into the secondary fibers. In the 20^th d of embryo(E20), the lens was maturation. By immunohistochemistry staining, the HIF-1α was highly expressed in the lens embryonic development. The expression was gradually promoting from E10 to E16, then reducing. The lens epithelium expressed more HIF-1α than fibers. The highest mean density was at E16, the lowest at E20. The difference was significant among of the 6 groups(P<0.0001). The E10 group was combined with the E12 group, the E14 group was combined with the E16 group, showing no significant difference(P>0.05). The other groups were compared with each other, finding significant difference(P<0.05). By the real-time PCR, HIF-1α mRNA was highly expressed in the lens development, and was different at different time. The expression of HIF-1α mRNA was increasing from E10 to E16, then descending at E18 and E20. The expression of HIF-1α mRNA was the highest at E16, the lowest at E20. The difference was significant among of the 6 groups(P<0.0001). The E10 group was combined with the E12 group, showing no significant difference(P>0.05). The other groups were compared with each other, finding significant difference(P<0.05).

CONCLUSION:The lens of Wistar rats differentiate from the E10 when the vesicle formed through the embryo phase. The lens is basic mature before birth. The HIF-1α in the lens embryonic development is highly expressed and changeful. The varies of HIF-1α expression is depend upon rat embryo development indicating that HIF-1α might participate in the process of development of rat lens.]]> 2014/10/31 11:03:29 Die Hu,Xu-Xia Meng,Hai-Tao Wang and Zhi-Ying Yu 205true METHODS: The third generation of cultured aged bovine RPE cells was cultured respectively with culture medium, containing 200μg/mL photoreceptor outer segment(POS)and 400μg/mL POS. Then the apoptosis rates of RPE cells after 24, 48 and 96h were detected with flow cytometry instrument(AnnexinⅤ/PI double staining).

RESULTS: In blank control group, the apoptosis rates were very low at different time. After RPE cells were cultured with culture medium containing 200μg/mL POS for 24h, the apoptosis rate including early apoptosis and late apoptosis was 1.73%. As time going, the apoptosis rate increased rapidly and reached 65.92% after 96h. After RPE cells were cultured with culture medium containing 400μg/mL POS for 24h, the apoptosis rate was 4.02%. And it reached 91.34% significantly after 96h.

CONCLUSION: Phagocytic loads can induce the apoptosis of RPE depending the time and dose.]]> 2014/10/31 11:03:29 Juan-Mei Zhang,Jian-Feng Wu and Hong-Sheng Bi 204true METHODS: Sixty 14-day-old healthy SD rats were randomly divided into 4 groups including normal group, monocular deprivation(MD)group, normal saline(NS)group, levodopa(LD)group, 15 rats for each. All the rats except those in the normal group were performed monocular deprivation surgery to establish monocular deprivation animal models and were raised in normal sunlight to 45-day-old. Rats in the MD group and normal group were killed and then marked by immunohistochemical and immune protein printing, respectively. The expression of NMDAR1 in visual cortex of these two groups were measured by quantitative real time PCR method. LD and NS groups were administered with levodopa(40mg/kg)and normal saline for 28d. The expression of NMDAR1 was examined with the same method.

RESULTS:NMDAR1 expression in visual cortex of MD group was lower than that in normal group. NMDAR1 expression in visual cortex of LD group was higher than that in NS group.

CONCLUSION: NMDAR1 is associated with the plasticity of visual development. Levodopa may revert the reduction of expression of NMDAR1 caused by monocular deprivation. This mechanism may be related with NMDAR1 capacity of improving visual function whose site of action may lie in the parts of the visual cortex.]]> 2014/10/31 11:03:29 Xiao-Nan Sun,Jun Tao and Jin-Song Zhang 203true METHODS: Totally 150 male Japanses white rabbits were divided into blank group(group A), sham-operated group(group B), model group(group C), androgen control treatment group(group D), and total flavonoid of chrysanthemum treatment group(group E). The dry eye model was established with orchiectomy on group C, D and E. Rabbits in group E were treated with total flavonoid of chrysanthemum. Rabbits in group D were treated with androgen intramuscular injection. Rabbits in the group A, group B, group C was treated with normal saline. All rabbits were detected with Schirmer's Ⅰ test and tear break-up time(BUT). Fas, FasL were checked on immunohistochemistry.

RESULTS: The Schirmer's I test values of group E was significantly higher than that of group C(P<0.01)and the BUT value of group E was significantly longer than that of group C(P<0.01). The quantity of positive expression of Fas in glandular tube cell and acinar epithelial celland apoptosis cells of group E after treatment at 1, 3, 5mo were significantly lower than that of group C, cell population of the positive expression of FasL was obviously higher than that of group C(P<0.01).

CONCLUSION:The main component of chrysanthemum is flavonoid, which could significantly inhibit happening of dry eye in rabbit after androgen level lowered and lacrimal gland apoptosis and keep basic tears secretory volume and tear film stability.]]> 2014/9/22 14:18:21 Xiao-Lei Yao,Qing-Hua Peng,Qi-Lei Chen,Yong-Hua Tang and Qian Zhong 202true METHODS: Thirty-six 7-week old Sprague-Dawley(SD)rats were intraperitoneally injected with MNU(60mg/kg)and were put to death by dislocation of cervical vertebra 6, 12, 24h; 3, 7d after injection(6 per group), respectively. As a control, six rats were injected with phosphate buffer saline(PBS)5mL/kg and sacrificed on d3 after injection. The degree of photoreceptor apoptosis was detected by HE staining, terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling(TUNEL)and transmission electron microscope(TEM)in the right eyes. The mRNA expressions of rhodopsin and recoverin were detected different time after injection by Western blot and immunohistochemical method in the left eyes.

RESULTS: The dissolution of photoreceptor nucleus and apoptosis body were first perceived at 12h by TEM; most of cells at outer nuclear layer were presented positive reaction. The apoptotic index reached peak(29.7%±2.3%)at 24h which was coincided with the observation of TEM. The results of immunohistochemistry displayed that rhodopsin and recoverin were on a declining curve with time extension. Furthermore, the results of Western blot indicated that rhodopsin had dramatic decline at 6h after injection(P<0.05), and extremely significant difference comparing to control group after 12h(P<0.01); while recoverin dramatic declined at 12h, and extremely significant difference after 24h(P<0.01).

CONCLUSION: 60mg/kg MNU intraperitoneally injection one-time may specifically induce photoreceptor apoptosis, The mechanism of down-regulation of rhodopsin and recoverin may be related to the selected apoptosis of photoreceptors.]]> 2014/9/22 14:18:21 Wei Jin,Yi-Qiao Xing,Hai-Feng Mei,Wen-Jun Wang and An-Huai Yang 201true METHODS: Preparation of optic nerve damage in 70 rats was randomly divided into normal group(10 rats), resveratrol treatment group(experimental group 30 rats)and PBS buffer control group(30 rats). The experimental group and control group was further divided into 3 subgroups(each group 10 rats), respectively. After 7, 14, 21d injected resveratrol or PBS, optic nerve injury were observed, then the rats were sacrificed. Retina was segregated; the surviving retinal ganglion cell(RGCs)was counted. Dissection of optic nerve, cholesterol content of them were tested; RT-PCR was used to detect mRNA expression of SIRT1, SREBP2 and HMGCR; Western blot assay was used to test the protein expression levels of SIRT1, cholesterol regulatory element binding protein 2(SREBP2)and HMGCR.

RESULTS: The numbers of RGCs and cholesterol levels of rat model with optic nerve injury decreased significantly(P<0.01). The mRNA and protein expression levels of SIRT1, SREBP2 and HMGCR were all decreased in a time-dependent manner(P<0.05). Three components of the three time points, with time injuries were aggravated, and the extent of damage was significantly reduced in the treatment group compared with the control group. But in resveratrol treatment group, the cholesterol levels and mRNA or protein expression of SIRT1, SREBP2, HMGCR in optic nerve were significantly restored in a time-dependent(P<0.05). The number of surviving RGCs restored significantly in resveratrol treatment group(P<0.01)in a time-dependent manner.

CONCLUSION: Up-regulating the expression of SIRT1, SREBP2 and down-regulating HMGCR by resveratrol could repair the injury of optic nerve through promoting the synthesis of cholesterol in neurons and retinal ganglion cells in the repair process. SIRT1 may be as a promising new target for treatment on optic nerve damage.]]> 2014/9/22 14:18:22 Yan Zhang,Hong-Yang Li and Yong-Mei Cao 200true METHODS: We designed and constructed B7-H1 gene adenovirus expression vector, and transfected DCs from mouse bone marrow, tested the phenotype and function of modified DCs, identificated its negative regulation to immune responses. The modified DCs were infected the sicked mice. And then the immunotherapeutic effect of modified DCs to TAO were tested.

RESULTS: B7-H1 gene adenovirus vector was constructed and transfected DCs from bone marrow. The titer of the recombinant adenovirus was 1.8×10⁹PFU/mL. B7-H1 gene modified DCs characteristics of regulatory DCs, could inhibit positive immune responses. The inhibition proceeding of TAO into mice infected modified DCs, was obviously prior to the control mice. The gene modified DCs, maybe become the new immunotherapy biological agent to thy TAO.

CONCLUSION: We constructed the expression of mouse B7-H1 gene adenovirus expressed vector successfully, transfected DCs,by vector have properties of regulatory DCs, inhibiting positive immune response and the occurrence and development of thyroid eye disease. Gene modified DCs, reveal potent to the treatment of thyroid eye disease.]]> 2014/9/22 14:18:22 Hua-Xin Chen,Bo-Zong Shao,Xuan-Chen Chen,Wei-Ming Zhou and Yi Zhang 199true METHODS: Thirty-six healthy female SD rats were selected and divided into four groups randomly, and the experimental group included three groups, which were individually injected 0.1mL 1.25, 5, and 10mU BTX-B solution on the right lacrimal gland; the control group was injected 0.1mL normal saline on the right lacrimal gland, then received Schirmer Ⅰ test(SⅠt)and examination of corneal fluorescein(FL)staining respectively at the 3, 7, 14 and 28d. The other 32 rats were selected and divided into two groups randomly, the rats in the experimental group were injected 0.1mL 1.25mU BTX-B solution on the right lacrimal gland and then five rats were randomly chosen to be removed lacrimal gland tissue respectively at the 3, 7, 14, 21, 42d. The Lacritin protein was detected in the qualitative and quantitative way by immunofluorescence and Western-blot, and the histopathological test was received by routine HE staining.

RESULTS: The three groups in the experimental group during the preparation of the model appeared that tear secretions decreased and corneal epithelium got damaged at 3d, but there was no significant difference for each other of two changes(P>0.05). The change was reached the peak at 7d and improved at 14d. The tear secretions returned to normal level at 28d, but the damage of corneal epithelium was still existed. The expression of Lacritin protein was only observed in acinar cells of both experimental group and control group, and the content of Lacritin protein in the experimental group decreased significantly. The decreasing situation appeared at 3d, reached the peak at 7d, improved at 14d, began to recover at 28d, and returned to the normal level at 42d.

CONCLUSION: Dry eye model of SD rats can be successfully established by lacrimal gland injection of 1.25mU BTX-B solution, with symptoms of dry eyes such as tear secretions reducing and corneal epithelial injury, which can provide experimental basis and foundation for the research on the pathogenesis and experimental treatment of dry eye. Lacritin protein only expresses in acinar cells of lacrimal gland, and the content of Lacritin protein has a synchronous change with tear secretions and degree of dry eye, which provides a new basis for perfecting the standards on evaluating the degree of dry eye.]]> 2015/8/27 11:19:42 Hai-Feng Zhu,Zhao-Qin Hao,Yu Cheng and Wei Gao 198true 2O₃)on the proliferation and apoptosis of on adenoid cystic carcinoma-2(ACC-2)cells and detect the expression of MDM2 gene from gene and protein level and to explore detailed mechanism of As₂O₃ inducing ACC-2 cells apoptosis.

METHODS: ACC-2 cells were cultured in vitro and divided into the experiment group and control group. Different concentrations of As₂O₃(2, 4, 6, 8μmol/L)were applied to cells in logarithmic growth phase at different time as experiment group, the control group was given the same amount of cell culture fluid, after added As₂O₃, the cells were cultured at different times, respectively. The effect of different As₂O₃ concentrations at each point time on inhibition and metamorphoses of ACC-2 cells was observed under inverted phase contrast microscope. Expression changes of MDM2 mRNA(24, 48h)and protein(24, 48, 72h)were determined by reverse transcription polymerase chain reaction(RT-PCR)and immunohistochemistry test(IHCT)respectively.

RESULTS: Cells shrinkage, nuclear chromatin condensation, apoptotic cells increased and the number of viable cells significantly reduced after being cultured with different concentrations of As₂O₃. The results of RT-PCR and IHCT were showed consistent the expression of MDM2 in experiment group decreased gradually with the increase of As₂O₃ concentrations and extension of action time, which was significantly different to that in the control group(P<0.05). Campared with each other, it was statistically significant between the different concentration and time of two groups(P<0.05). MDM2 expression was negatively correlated with concentration and time(r<-0.7, P<0.05), that was, it presented in dose- and time-dependent manner.

CONCLUSION: As₂O₃ has the inhibitory and apoptosis-inducing effect on ACC-2 cells, and it can downregulate the expression of MDM2 mRNA and protein in ACC-2 cell line. This may be the mechanism of As₂O₃ induced ACC-2 cells apoptosis.]]> 2015/8/27 11:19:42 Shuang-Shuang Wang,Tao Jiang,De-Wei Li,Xiao-Yan Tong,Xiao-Chuan Wang and Yu Zhang 197true METHODS: The DM model was established by a single injection of streptozotocin. Diabetic rats were intravitreal injected with adenovirus vehicle(DM group), or carried glucocorticoids receptor antisense(siGR group), or scrambled nucleotide(scRNA group). Control group(CON group)were normal SD rats intravitreal injected with adenovirus vehicle. Twelve weeks later, the density of retinal ganglion cell(RGC)and retinal thickness were detected by HE staining, and the expression of ROCK was observed by immunohistochemical staining and Western-blot.

RESULTS: Compared with CON group, glucocorticoid concentrations were significantly increased in DM group, siGR group and scRNA group(P<0.01, respectively). DM group and scRNA group showed reduction of RGC density and retinal thickness, and up-regulation of ROCK(P<0.01, respectively). While no alterations was detected between CON group and siGR group(P>0.05, respectively).

CONCLUSION: Inhibition of glucocorticoids in diabetes reverses the expression of retinal ROCK, RGC density, and retinal thickness.]]> 2015/8/5 15:43:41 Bo Zhang,Yu-Bo Wang,Wen-Qiang Liu and Xue-Zheng Liu 196true METHODS: MTT assay and cells scratch were adopted to observe hyperplasia of human umbilical vein endothelial cells(HUVECs)and cell migration induced by sphingosine-1-phosphate(S1P)after using FTY720 of different concentration. The effect of FTY720 on CNV induced by S1P in a rat corneal micropocket model was detected. 30SD rats were randomly divided into group A, group B and group C with 10 rats per group. S1P and 0μg, 5μg, and 20μg FTY720 controlled-released particles were implanted into the corneal stroma. The growth of CNV and having pathological examination on 12d after the operation was observed. Findings was analyzed by one-way ANOVA.

RESULTS: 10, 10², 10³, and 10⁴ nmol/L FTY720 and HUVECs co-incubate 72h could inhibit cell proliferation(P<0.01), 24h after the function of 10,100nmol/L FTY720, it could inhibit S1P-induced cell migration and the ability of restricting cell proliferation and cell migration was enhanced with increasing concentration of FTY720. On 12d, after rat corneal micropocket controlled-release particles was implanted into groups A, B, C, the CNV area were respectively 10.05±1.19, 6.59±0.95, 2.70±0.68mm²(F=145.155, P<0.01), group A and group B was statistically different and this was the same case between group B and group C(P<0.01).

CONCLUSION: FTY720 can inhibit S1P-induced corneal neovascularization.]]> 2015/7/1 9:46:00 Fan Zhong,Xiao-Zhen Ding,Wei-Zhong Yang,Zong-Yin Gao and Xiao-He Lu 195true METHODS: OCM-1 cells cultured on culture flask were divided into normal group and hypoxia group. Hypoxia group were divided into five groups: simple hypoxic group, and interference group, and negative control group, and positive control group, and liposome group. Normal group cells were cultured on DMEM culture flask with 10% FBS, 100U/mL penicillin, 100μg/mL streptomycin as well as high concentration of glucose. The cells were maintained at 37℃ in a humidified 5% CO₂ incubator. Cells in good condition were selected for experiment. For hypoxia group, chemical hypoxia inducer CoCl₂ was added into nutrient medium at the concentration of 100μmol/L to simulate hypoxia microenvironment. We designed and synthesised siRNA(siRNA+negative control+positive control), the target sequences of the HIF-1α to transfect hypoxic malignant melanoma of the choroid cell. SiRNA including HIF-1α siRNA, β-actin siRNA and negative control group synthesized in vitro transfected hypoxic OCM-1 cell through Lipofectamine2000. The expression of HIF-1α, MMP-2 gene and the protein were detected by RT-PCR and Western blot.

RESULTS: Compared with the normal group, the expression of HIF-1α mRNA was not obviously changed(P>0.05), but the expression of HIF-1α protein and MMP-2 mRNA protein was significantly higher(P<0.05). Compared with the other hypoxia groups,β-actin mRNA expression of positive control group decreased(P<0.05), which proved successful transfection. The expression of HIF-1α mRNA and the expression of its protein and both MMP-2 mRNA and its protein was significantly lower(P<0.05). The negative control group, liposome control group had no significant difference in the detection of factors(P>0.05).

CONCLUSION: Hypoxia status may upregulate the HIF-1α in OCM-1 cells by increasing the expression of protein. Hypoxia can also inactivate MMP-2, resulting in upregulation of MMP-2 RNA and the expression of its protein. The expression of HIF-1α and MMP-2 mRNA can be down-upregulated by transfecting OCM-1 with HIF-1α siRNA.]]> 2015/7/1 9:46:01 Fu-Xiang Yuan,Ting Zhang,Zhan-Yu Zhou,Liang-Yu Wang,Juan Zhao,Shuang Wang and Fa-Yan Lu 194true 2O₂).

METHODS:ARPE-19 cells were treated with different concentrations of α-Mangostin and H₂O₂.The effect of α-Mangostin and H₂O₂ respectively on cell activity was detected by CCK8. ARPE-19 cells were pretreated with different concentrations of α-Mangostin for 24h before they were administrated with 200μmol/L H₂O₂for another 24h.Then the changes of cell activity were observed. The expression of reactive oxygen species(ROS)level was detected by flow cytometry(FCM)and the expression of NF-κB protein was measured by Western blot analysis.

RESULTS:CCK8 examination results showed that: within 0～12μmol/L, α-Mangostin had no damage effects on cell activity. When the concentration of 16μmol/L, cell viability began to decrease(P<0.05).And α-Mangostin pretreatment gradually increased cell viability of ARPE-19 induced by H₂O₂ when the concentrations of α-Mangostin were within 0～16μmol/L. ROS results showed: the expression of ROS level significantly increased after H₂O₂ induced(P<0.05); 8 and 12μmol/L α-Mangostin pretreatment down-regulated ROS expression of ARPE-19 induced by H₂O₂(P<0.05). Western blot results showed that the expression of NF-κB protein after H₂O₂ induced increased(P<0.05); 12μmol/L α-Mangostin pretreatment up-regulated NF-κB of ARPE-19 induced by H₂O₂(P<0.05).

CONCLUSION: H₂O₂ induced oxidative damage in RPE cells by decreasing cell viability and increasing the expression of ROS level. α-Mangostin can protect RPE cells from the injury of H₂O₂, the mechanism may be related to the clear of ROS and the activation of NF-κB.]]> 2015/6/1 16:06:21 Tu Su,Yuan Fang,Ping Xie,Song-Tao Yuan,Wen Fan,Yi-Dan Xu and Qing-Huai Liu 193true METHODS: Lipoglycans was extracted by the Triton X-114 phase partitioning. Lipoglycans from Che were purified, by successive detergent and phenol extractions. Lipoglycans were separated by gel filtration on a Sephacryl 200 column and Sephacryl 100 column in series, followed by extensive dialisis. Purified Lipoglycans(50μg/mL)were added into culture medium to stimulate primary human corneal epithelial(HCE)cells. Cells and supernatant were collected at 0, 6, 12, 24h after the stimulation. The IL-6 and IL-8 expression at mRNA level was assayed by using real time RT-PCR and the secreted IL-6 and IL-8 in the supernatants was measured by ELISA. Immunochemistry was used to detect the expression and location of NF-κB in HCE cells.

RESULTS: After the treatment of Lipoglycans, the expression of IL-8 and IL-6 at mRNA level obviouly increased within 12h, and reached peak level at 6h(IL-8 was 36.8 times that of the blank control, and IL-6 was 32.7 times). Compared with the blank control group, the expression of IL-8 at protein level in the supernatant increased 2.8 folds at 6h(P>0.05), 13.4 folds at 12h(P<0.05), and 200.7 folds at 24h(P<0.05), and the expression of IL-6 also incresed 3.6 folds at 6h(P<0.05), 6.1 folds at 12h(P<0.05), and 7.0 folds at 24h(P<0.05), which was similar to changes in the positive control group(situmulated by LPS). In the blank control group, NF-κB was localized in the cytoplasm, while in lipoglycans treated group, NF-κB was activated and translocalized to the nucleus in HCE.

CONCLUSION: Lipoglycans from Che can induce HCE cells to produce inflammatory factors(IL-6 and IL-8), and its signal transduction pathway probably is mediated by NF-κB.]]> 2015/6/1 16:06:21 Chun-Zhou Tang and Huai-Jin Guan 192true METHODS:Real time q-PCR was used to measure the expression of miR-181 in the anterior lens capsules of age-related cataract and human lens epithelial cell apoptosis model. miR-181 mimic and inhibitor were transfected using Lipofectamine 2 000 to regulate the expression of miR-181, and then Real time q-PCR was used to verify transfection efficiency. Flow cytometry was used to detect the change of cell apoptosis rate.

RESULTS: Compared with control group, the expression of miR-181 was significantly higher in both the anterior lens capsules of age-related cataract and human lens epithelial cell apoptosis model; the relative expression of miR-181 in lens epithelial cells transfected with miR-181 mimic was increased, whereas decreased in cells transfected with miR-181 inhibitor; the apoptosis rate of cells transfected with miR-181 mimic was increased, while reduced in miR-181 inhibitor group. Each result was statistically significant(P<0.01).

CONCLUSION: High expression of miR-181 is detected in anterior lens capsule of age-related cataract. miR-181 might play a certain role in the pathogenesis of cataract via promoting human lens epithelial cell apoptosis. miR-181 probably becomes a new approach for the nonoperative treatment of cataract, but the concrete mechanism still needs to be further studied.]]> 2015/5/5 16:34:50 Yu Qin,Jiang-Yue Zhao,Wen-Ting Luo,Jing Li,Jia Liu and Jin-Song Zhang 191true 2O₂)-induced oxidative damage to human lens epithelial cells.

METHODS: Sub-culture human lens epithelial cells preprocessed with different concentrations of melatonin for 12h and then 100 μmol/L H₂O₂ for 24h. The impact of melatonin on H₂O₂-induced lens epithelial cell viability was detected by MTT assay, rate of apoptosis was detected by flow cytometry instrument and activity of apoptosis-related factors, Caspase-3 and Caspase-9, were detected by colorimetric method.

RESULTS: MTT assay showed that melatonin had no effect on the activity of lens epithelial cells, and the drug can inhibit the decrease of H₂O₂-induced cell activity, as well as flow cytometry showed that melatonin can inhibit H₂O₂-induced apoptosis. In addition, melatonin can also reduce H₂O₂-induced Caspase-3 and Caspase-9 activity in lens epithelial cells, and their activity decreased with effect of melatonin along with extending time.

CONCLUSION: Melatonin can obviously inhibit H₂O₂-induced apoptosis of human lens epithelial cells, which provide reliable experimental basis for drug on treatment of cataract.]]> 2015/5/5 16:34:50 Tian-Rui Dong,Ping Liu and Jin-Hong Ni 190true METHODS:Both hMSCs and RPE cells were co-cultured in a transwell system. The experiment was divided into four groups: 21% O₂ with 5.56mmol/L glucose(control group, A), 21% O₂ with 30mmol/L glucose(hyperglycemia and normoxia group, B), 5% O₂ with 5.56mmol/L glucose(normoglycemia and hypoxia group, C)and 5% O₂ with 30mmol/L glucose(hyperglycemia and hypoxia group, D). Cell Counting Kit-8(CCK-8)was used to detect the proliferation of RPE cells in each group at 12, 24 and 48h respectively. Flow cytometry was performed to observe apoptosis of RPE cells at 24h. Additionally, we assessed migration capabilities of RPE via transwell assay under the condition of hyperglycemia and hypoxia by co-culturing of hMSCs.

RESULTS:In this co-culturing system, at 12, 24 and 48h, group B(1.61±0.41, 1.80±0.34; 1.91±0.35), C(1.34±0.46, 1.94±0.40, 2.14±0.41)and D(1.98±0.47, 2.26±0.42, 2.55±0.40)showed significantly higher proliferation rate than group A(0.92±0.45, 1.27±0.32, 1.59±0.41, P<0.05). The migration capabilities of RPE in group B(149.5±9.19), C(140±9.90)and D(170.5±7.78)increased dramatically compared with group A(114.5±7.78, P<0.05)at 24h, whereas there was no significant difference of apoptosis ratio among four groups(P>0.05).

CONCLUSION:By coexistence with hMSCs, the synergy of hyperglycemia and hypoxia can improve migration and proliferation of RPE cells, and have no effect on apoptosis of RPE cells within short period.]]> 2015/4/8 9:56:56 Jie-Jing Yan,Hai-Yan Wang,Yu-Sheng Wang,Fan Gao,Na Li and Peng Zhang 189true METHODS: Human lens epithelial cells B3(HLE B3)were treated with 1.5mg/mL AGEs-BSA as the experimental groups cultured by fetal bovine serum of volume fraction 10% dulbecco modified eagle medium(DMEM)and bovine serum albumin(BSA)was added at the same concentrations as the negative control. The level of reactive oxygen species(ROS)was evaluated. Cells were collected at 0, 1, 2, 3, 4d and total RNA or protein was extracted. TTase mRNA levels were detected by qRT-RCR. TTase expression was detected by Western blot and its activity was measured.

RESULTS: Compared with the control group,AGEs-BSA up-regulated the expression of ROS(P<0.01), ROS content increased in a time-dependent manner. BSA had no effects on ROS expression. The expression of TTase increased after treatment with AGEs-BSA for 1d, peaked at 2d(nearly 5.06-fold increase, P<0.01), then decreased gradually. No change was observed between BSA and control group(P>0.05). Similarly, TTase activity peaked at 3d(nearly 2.01-fold increase, P<0.01). Western blot test found that TTase protein expression was increased gradually, starting from the 3d TTase expression was reflected that there was statistically significant difference compared with control group(P<0.05).

CONCLUSION:AGEs-BSA significantly increases the production of ROS in human lens epithelial cells, and it then induces the oxidative stress which may promote the expression of TTase and enhances the activity of TTase.]]> 2015/4/8 9:56:56 Xu Wang and Hong Yan 188true METHODS: Twenty normal SD rats were randomly divided into two groups: control group(n=10)and atropinization(experimental)group(n=10). All the left eyes were selected as the experimental eyes, and the right eyes served as the normal eyes. The left eyes in atropinization group was produced by 1% atropine, 3 times a day and the right eyes in control group was treated with normal saline, 3 times a day. The flash visual evoked potentials(F-VEP)and retinoscopy refraction of the rats' both eyes were detected at five time points: 0, 7, 14, 21, and 28d after atropinization, respectively. After 28d, six rats were randomly selected from both groups and each group had three rats. The expression of the c-fos mRNA was observed in both visual cortexes. Another six rats were chosen for the same test after 2d dark environment with 2h light later. The expression of c-fos mRNA was detected again.

RESULTS: After 14d anisometropia was observed in experimental group, the difference was 3.9D(P<0.05), F-VEP P1 wave of the rats left in experimental group was reached to 88.9±1.889ms at 21d, there was statistical difference compared with the right eye(P<0.05). After 28d, c-fos mRNA expression in the left visual cortex of rats in the experimental group was higher than that of the right side, but there was no significant difference. But when underwent 2h light stimulation after in the darkroom 2d, the c-fos mRNA expression in in the left visual cortex of rats in the experimental group was 5 times higher than that of the right side, there was a statistically significant difference(P<0.05).

CONCLUSION: In the critical period of visual development, monocular chronic atropine in rats can form anisometropia, may delay the transmission of the optic nerve, hinder the normal development of the visual cortex. Monocular atropinization in rats can be used as the model of anisometropia.]]> 2015/4/8 9:56:56 Ya-Zhen Wu,Yong-Xin Xing and Hong Yan 187true +CD25⁺T cells and expression of FOXP3 mRNA in peripheral blood mononuclear cell(PBMC)from patients with proliferative diabetic retinopathy(PDR)and observe its clinical significance on pathogenesis of PDR.

METHODS: Flow cytometry was used to analyze the proportion of CD4⁺ CD25⁺ T cells in PBMC with PDR and normal control, the expression of FOXP3, IL-17, CTLA-4 mRNA in patients with PDR, diabetes and normal control were detected by Q-PCR. The clinical data of glycosylated hemoglobin(HbA1c), age, blood lipid, renal function and the degree of their relevance were analysed by Wilcoxon rank and inspection, Linear correlation analysis.

RESULTS:CD4⁺ T cells decreased, CD4⁺CD25⁺T with no significant difference, FOXP3 and CLAT-4 decreased in PDR, IL-17 increased in PDR patients. CD4⁺T cells associated with age, the level of CD4⁺CD25⁺T was positive correlation with the patient's serum creatinine(Cr), but had no significant with age, HbA1c, blood lipid, Cr and urea nitrogen(BUN). HbA1c, triglyceride(TG), cholesterol, low density lipoprotein(LDL)were higher than normal value in PDR patients.

CONCLUSION: CD4⁺ CD25⁺ T cells may be involved in the pathogenesis of PDR. In addition, abnormal blood lipid indicate that PDR may be associate with blood lipid level.]]> 2015/3/9 10:14:41 Wen-Yi Wu,Luo-Sheng Tang,Hui-Hui Chen and Fang Li 186true METHODS:Rhesus choroid-retinal vascular endothelial cells(RF/6A)were cultured under normoxic and hypoxic conditions in vitro and divided into the normoxia group, the hypoxia control group(transiently transfected with the vector plasmid)and the treated group(transiently transfected with the CCR7siRNA recombinant plasmid). Plasmids were transiently transfected in RF/6A cells using Lipofectamine^TM2000(LF2000). RF/6A cells proliferation and apoptosis detected by CCK8 and flow cytometry and the protein and mRNA expression of CCR7 and VEGF were measured by Western blot and RT-PCR.

RESULTS:The cell growth rate became slower and apoptosis rate became increased in the hypoxia group than that in the normoxia group, and in the treated group than that in the normoxia and hypoxia control groups(all P<0.05). Compared with the normoxia group, there were high protein and mRNA expression of CCR7 and VEGF in the hypoxia control group, the differences had statistical significance(t_CCR7protein=3.38, t_VEGFprotein=4.75, t_CCR7mRNA=4.27, t_VEGFmRNA=5.34, all P<0.05). And there was an obvious positive correlation between the expression of CCR7 and VEGF(r_protein=0.71, r_mRNA=0.83, all P<0.05). The protein and mRNA expression of CCR7 and VEGF were obviously decreased in the treated group compared with the normoxia and hypoxia control groups(all P<0.05).

CONCLUSION: CCR7 can upregulate the expression of VEGF in REC under hypoxia. CCR7- VEGF signaling pathway may have potential function in the retinal neovascularization(RNV), and CCR7siRNA may provide an effective method for RNV.]]> 2015/3/9 10:14:41 Yu Di,Xiao-Long Chen and Ai-Yuan Wang 185true METHODS: Forty Sprague Dawley rats(SD rats)were intraperitoneally injected with NaIO₃ (30g/L, 100mg/kg)to establish the retinal degeneration models(postnatal 28d). These rats were devided into 4 groups. Group A was not injected, group B was injected with BMSCs, group C was injected with BMSCs and ChABC, and group D was injected with phosphate buffer saline(PBS). After 28d, subretinal injection were applied. Hematoxyln-eosinstaining(HE), tunel and immunohistochemistry were performed at 21d after subretinal injection.

RESULTS: Photoreceptor number and photoreceptor apoptosis rate of B and C groups were more than those of A and D groups, and there was significant difference statistically(P<0.05). Photoreceptor number and photoreceptor apoptosis rate of group B were compared with those of group C, and there was no statistical significance between B and C groups(P>0.05). Glial fibrillary acidic protein(GFAP)was expressed by BMSCs after intraocular injection.

CONCLUSION: BMSCs and ChABC injected into subretinal space may alleviate photoreceptor apoptosis so as to protect retinal photoreceptor cells in degenerated rats.]]> 2015/3/9 10:14:41 Xiang-Rong Zheng,Lin Liu and Peng-Fen Gao 184true METHODS: Recombinant PGC-1α protein was added to HRVEC, and no recombinant PGC-1α protein was added to HRVEC as control group. After 24h of incubation, two groups of cells were then placed into a normoxic(20% O₂)or hypoxic(1% O₂)environment for another 16h. The expression of VEGF mRNA and protein were detected by real-time PCR, ELISA and immunofluorescence cytochemistry.

RESULTS: VEGF mRNA and protein levels in the cells were significantly increased by recombinant PGC-1α protein both under normoxia and hypoxia conditions as compared with control groups(P<0.01).

CONCLUSION: PGC-1α can upregulate the expression of VEGF in HRVEC under normoxia and hypoxia conditions.]]> 2015/1/30 15:09:34 Jian Jiang,Xiao-Bo Xia and Lu Zhang 183true 1(TGF-β₁)expression in vitro culture rat corneal stromal cells.

METHODS: Corneal stromal cells from 8 to 10wk SD rats were isolated, cultured and treated with different concentrations of PFD 0mg/mL(control group), 0.15mg/mL(experimental group Ⅰ), 0.3mg/mL(experimental group Ⅱ), 1mg/mL(experimental group Ⅲ)for 48h. CCK-8 assay was performed to assess cell proliferation, while immunocytochemistry and Western Blot were used to detect the expression of ki-67 and TGF-β₁ expression, respectively.

RESULTS: Compared with control group, PFD significantly inhibited the proliferation in a dose-dependent manner(all P<0.05), so was protein expression of ki-67. PFD significantly down-regulated the expression of TGF-β₁ in a dose-dependent manner(P<0.05).

CONCLUSION: Pirfenidone can significantly inhibit the proliferation of rat corneal stromal cell by down regulating TGF-β₁ expression, therefore, it has potential prospect in lightening the corneal wound healing reaction.]]> 2015/1/30 15:09:35 Jun-Jie Chen,Gong-Fa Wu,Jun-Shan Lin,Yu-Ting Zeng and Qi-Ting Huang 182true METHODS:Four effective components of Qingguang'an were used in groups D, E, F, G and H after glaucoma surgery, compared with group A(blank), group B(model)and group C(MMC)to observe the effect of elastic fiber, MMP-7, TIMP-1 in scarring filtration canal.

RESULTS:Compared with the preoperative basic IOP and 2d, 1, 2, 4wk postoperative IOP of groups C, E and H, the IOP of three group rose up slower than other groups, and kept the lowest data at 28d. There was significant difference compared with the rest of A, B, D, F, G groups(P<0.05). The area and density of elastic fiber in surgery group were significantly different with that of black control group(P<0.05), but there were no statistical differences between groups C and H, groups C and F, groups H and E(P>0.05). The difference was statistically significant among other groups(P<0.01).

CONCLUSION:The scarring area of filtration canal after glaucoma surgery is the major reason which lead to the failure of surgery. Qingguang'an effective group 2, Qingguang'an granules and MMC could reduced the scar tissue by restrained the elastic fiber, TIMP-1 and increased the MMP-7. By observing the experimental results that both Qingguang'an effective group 2 and Qingguang'an granules could restrained the scarring area of filtration canal, the effects were unbiased, Qingguang'an granules group is better than effective group 2.]]> 2014/12/30 15:26:03 Yuan-Bi Li,Qing-Hua Peng,Xue-Si Huang,Xiao-Liu Chen and Han-Yu Tan 181true in vitro cultured lens epithelial cells(LECs).

METHODS: In vitro cultured infant human lens tissue HLE B-3 immortalized cells were distributed randomly divided into six groups. Each group was administered with free Zeb 50μmol/L(ZebF1 group), 100μmol/L(ZebF2 group), Zeb -loaded MePEG-PCL NPs 50μmol/L(ZebNP1 group), Zeb -loaded MePEG-PCL NPs 100μmol/L(ZebNP2 group), MePEG-PCL empty NPs(NPs group)or blank medium(group C)respectively. A tetrazolium dye assay(MTT)test and modified MTT test were performed to determine cell viability and cell attachment. DNA ladder was used to detect the cell apoptosis.

RESULTS: Determined by MTT colorimetric method: Cell proliferation rate of LECs were suppressed by all Zeb administration groups in a concentration-time dependent manner(P<0.05). Compared with the free Zeb groups, the viability of LECs were suppressed more effectively by the same dose of Zeb loaded MePEG-PCL NPs after 24, 48, 96h(P<0.05). Determined by improved MTT colorimetric method: The attachment of LECs were decreased in all Zeb administration groups, Zeb loaded MePEG-PCL NPs had better effect on suppressing the attachment of LECs than the free Zeb groups with same dose(P<0.05). The DNA ladder confirmed that after administration of 96h, group C, NPs group and ZebF1 group showed no DNA fragment, however the DNA fragment were performed in ZebF2, ZebNP1, ZebNP2 groups and displays the trend of ZebNP2> ZebNP1>ZebF2(P<0.05).

CONCLUSION: Zeb loaded MePEG-PCL NPs had better effect on suppressing the viability and attachment of in vitro cultured LECs than the free Zeb groups, as well as enhancing the apoptosis.]]> 2014/12/30 15:26:04 Si-Wei Liu,Qun Wang and Qian-Yan Kang 180true METHODS:Real-time PCR was applied to detect the expression of miR-218 in human RB and the corresponding tumor-adjacent tissues to analyze the relation between miR-218 and clinic pathology characteristics. The artificial miR-218 was transiently transfected into human Y79 cells in vitro. The proliferation and apoptosis of the cells were detected by MTT assay and flow cytometry, respectively. The expression level of Bmi-1 mRNA and protein were determined by RT-PCR and Western blot.

RESULTS:The expression level of miR-218 in RB tissues was significantly lower than that in the adjacent tissues, and it was associated with optic nerve infiltration and differentiated degree. Over-expressed miR-218 in Y79 cells suppressed cell proliferation and promoted cell apoptosis, and down-regulated mRNA and protein expressions of Bmi-1.

CONCLUSION:The expression of miR-218 in RB tissues is significantly lower than that in tumor-adjacent tissues.MiR-218 could inhibit RB cell proliferation and induce apoptosis by down-regulating the expression of Bmi-1.]]> 2015/11/27 10:32:21 Lin-Sheng Yang,Li-Lun Wang and Qing-Wei Du 179true METHODS:Human retinal vascular endothelial cells were cultured in low glucoseor high glucose environment.The MTT assay was used to detect the proliferation of each group to study the effects of resveratrol on the proliferation of human retinal vascular endothelial cells in high glucose environment. Detection of Sirtuin 1(SIRT1)expression and acetylated high mobility group box-1 protein 1(HMGB1)was performed by western-blot and coimmunoprecipitation.

RESULTS:Resveratrol inhibited the proliferation of human retinal vascular endothelial cells in high glucose in a dose-dependent manner.High glucose decreased SIRT1 expression andincrease the degree of HMGB1 deacetylation, which can be reversed by resveratrol.

CONCLUSION:Resveratrol may reverse proliferation of retinal vascular endothelial cells in high glucose through SIRT1-HMGB1 pathway.]]> 2015/11/27 10:32:21 Yan Wei,Hong Li,Xiao-Qing Su and Zhen-Guo Yan 178true METHODS:Thirty two-week-aged guinea pigs were randomly divided into three groups, group A, group B and group C. Another five guinea pigs(10 eyes)were chosen as control group without any treatment. Concave lenses were worn by the 30 guinea pigs on either side of their eyes. After 6, 15 and 30d, the lenses were removed and optometry and axial length were used to make sure the myopia had formed. The retinal pigment epithelial cells of each group were cultured and passaged in vitro using enzymatic digestion method. The cultured cells at 3～6 generation was detect by immunocytolchemistory, Real-Time PCR and Western blotting to detect the expression of bFGF.

RESULTS:The expression of bFGF was located in the cytoplasm and nucleus. The results of immunocytolchemistory, Real-Time PCR and Western blotting showed that the expression of bFGF were detected at anterior and posterior part of eyes from experiment group and control group, the expression in experiment group at both part was significantly lower than those in control group(P<0.05). The positive percentage of expression of bFGF in experiment group decreased with the induced time(P<0.05). The expression of bFGF from anterior part of both experiment group and control group was as the same as that from posterior part of themselves(P>0.05).

CONCLUSION:The expression of bFGF from anterior and posterior part of experiment group was lower than those of control group.]]> 2015/10/30 17:17:32 Bo-Yu Chen,Chao-Ying Wang,Wei-Yi Chen,Ying-Qing Liu and Lan Hao 177true -/-fibroblasts on thyroid-associated ophthalmopathy.

METHODS:Optimal shRNA interference expression plasmid of mouse orbital fibroblasts TLR4 gene was designed, built, and screened. Then the best shRNA was introduced into lentiviral expression vector by Gateway method and recombinant lentiviral vector was used to infect mouse orbital fibroblasts. Its capability of negative regulating the immune inflammatory response was researched. At last the method of fibroblast TLR4 gene silencing in the model mice of thyroid-associated ophthalmopathy was used, the in vivo therapeutic effect was observed.

RESULTS: ShRNA sequences with the best effect of gene silencing were selected and introduced into lentiviral vectors(virus titer was 1.5×10⁶TU/mL). Balb/c mice orbital fibroblasts transfected lentivirus could negatively regulate the immune response, inhibit immune inflammatory response. The proceeding of thyroid-associated ophthalmopathyof the mice transfected TLR4^-/-recombinant lentivirus was obviously prior to that of the control mice.

CONCLUSION: Mouse fibroblast TLR4^-/- siRNA lentiviral vectors are successfully obtained, which has the favourable inhibitory effect on immune inflammatory responses. The recombinant lentivirus could protect the proceeding of thyroid-associated ophthalmopathy, therefore the TLR4 expression interference is a novel potential target for thyroid-associated ophthalmopathy.]]> 2015/10/30 17:17:32 Wen-Ying Wang,Wei-Ming Zhou,Yi Zhang and Hua-Xin Chen 176true 1(TGF-β₁).

METHODS: Firstly, the effect of TSA at different concentrations(200, 400, 600 and 800nmol/L)on HTFs viability after 24h was detected using MTT proliferation assays. Then, the effect of TSA at 400nmol/L and 600nmol/L mixed with 5ng/mL TGF-β₁ on HTFs viability after 24h were investigated using MTT proliferation assays. Furthermore, the mRNA and protein expression of collagen Ⅰ in HTFs after treatment with TSA at different concentrations(400, 600nmol/L)mixed with 5ng/mL TGF-β₁ as well as 600nmol/L TSA were detected by RT-PCR and Western-blot.

RESULTS: Compared with control group, the results of MTT showed that HTFs viability decreased significantly after treated with TSA at 400, 600 and 800nmol/L(P<0.05). The HTFs proliferation induced by TGF-β₁ could be attenuated by TSA at 400 and 600nmol/L(P<0.05). The results of RT-PCR and Western-blot confirmed that TSA at 400 and 600nmol/L had reversal effect on up-regulated gene transcription and protein expression levels of collagen Ⅰ induced TGF-β₁.

CONCLUSION:TSA can inhibit the HTFs proliferation induced by TGF-β₁ and attenuate gene transcription and protein expression of collagen Ⅰ.]]> 2015/9/25 17:05:18 Xiao-Yan Li,Ying Deng and Cheng Pei 175true METHODS: Neonatal C57BL/6J mice were divided randomly into three groups: normoxic control group, physiological saline injection of OIR group and adiponectin injection of OIR group. The mice of the latter two groups were exposed to 75%±2% oxygen from 7d(P7)～P12 to induced OIR. Recombinant APN(rAPN)was injected intraperitoneally(i.p., 3.0μg/g)in a mice model of OIR from P7～P15. Another set of mice model of OIR were received a similar treatment with physiological saline. All eyes were collected at P17. The right eyes were whole mounted and stained with Lectin to observe central retinal avascular area and the growth of pathological neovascularization; The left eyes were performed histopathological cross sections stained with HE to analyzed the histopathological changes in the retina. The eyes were enucleated to assess the levels of reactive oxygen species(ROS)and NO. The protein expression of iNOS, nNOS, eNOS were detected by Western-blot.

RESULTS: The central retinal avascular area, neovascular area were markedly decreased after the APN injection compared with physiological saline injection of OIR group(t=7.304, P<0.01; t=2.654, P<0.01). Compared with physiological saline injection of OIR group, the levels of ROS were lower(t=13.349, P<0.01), the levels of NO were higher(t=3.023, P<0.01), the expression of iNOS were decreased(t=5.112, P<0.01), the expression of eNOS were decreased(t=7.421, P<0.01). nNOS expression had no significant difference(t=1.074, P>0.01).

CONCLUSION:The realtus demonstrate that APN can promote physiological NO by acting endogenous eNOS, while suppress ROS/RNS generation and play a protective role in retinal vessels in OIR process.]]> 2015/9/25 17:05:18 Qiao-Ping Zhu,An-Ming Xie and Yan-Fang Hao 174true METHODS:Twenty-five Wistar rats were randomly divided into five groups: sham operation group, 1d, 3d, 7d and 14d after tractive optic nerve injury group. The model groups pulled the left optic nerve with lateral tensiometer; the sham operation group only exposed the optic nerve but not pulled. The right eyes of each group were served as normal control eyes. The RGCs density of the five groups was observed by fluorogold retrograde labeling.

RESULTS:In the normal control group, the RGCs labeled by the fluorescent gold were round or oval, clear boundary, no obvious fluorescent dye leakage and partially visible cell processes. However, in the optic nerve traction groups, the number of RGCs decreased with time increasing and the cell distribution was not uniform. Lots of fluorescent leakages and microglial cells were observed. Compared with the normal control group, there was no significant difference in the morphology and density of RGCs of sham operation group(P>0.05). In 1d, 3d, 7d and 14d after traction of the optic nerve groups, the number of RGCs were reduced progressively and the density of RGCs of the left eye was significantly lower than that in the normal control group(P<0.01). The survival rates of RGCs in the groups of 1d, 3d, 7d and 14d after optic nerve traction were(78.94±0.92)%,(60.07±0.90)%,(38.92±1.42)% and(17.31±0.97)% respectively.

CONCLUSION:The transverse quantitative traction method can establish a model of easily quantifiable optic nerve injury, which can provide a powerful tool for further study on the treatment of optic nerve injury.]]> 2016/8/22 9:57:56 Xin-Yi Gu, Jian Zhou, Peng-Bo Zhao, Ai-Wei Liu, Shan-Shan Shang, Xiao-Ling Yan, Lu-Hua Wu and Yan-Ting Xia 173true METHODS: Twenty-five Sprague-Dawley male rats were taken as experimental subjects. Random 5 of them were used as control group. The remaining 20 were induced to be diabetic mellitus by a single intraperitoneal injection of streptozotocin(STZ)and divided into 4 groups randomly: diabetes control group, 0.01% Salvianolic acid B treated group, 0.05% Salvianolic acid B treated group and 0.1% Salvianolic acid B treated group. In the Salvianolic acid B treated groups, the corresponding concentration of Salvianolic acid B eye drops were used 3 times a day until the animal was sacrificed, and then detected rats blood glucose and body weight, observed sensation and transparency of the cornea once a week. After the building completion, all rats in each group were sacrificed in 12^ndwk, and then observed the corneal tissue structure change, and in rat serum and tissue of malondialdehyde(MDA)content and activity of superoxide dismutase(SOD).

RESULTS: The blood glucose of all rats was in high level(>16.7mmol/L), early rise and later fall in weight. In the diabetic control group, the corneal sensation and transparency of the cornea were decreased. The content of MDA in the corneal tissue was significantly increased. The activity of SOD in the corneal tissue was significantly decreased(P<0.05). Through the course of disease, the above change was obvious day by day. Compared with the diabetic control group, the sensation and transparency of the cornea were improved significantly. The content of MDA in the corneal tissue was significantly decreased. The activity of SOD in the corneal tissue was significantly increased(P<0.01).

CONCLUSION: Salvianolic acid B can reduce the corneal lesions of diabetic rats, the mechanism may be related to Salvianolic acid B, remove superoxide anion, protect cells from damage of resisting oxidative stress.]]> 2016/8/22 9:57:56 Sheng-Hui Wu, Yan-Feng Wang, Xiao-Qin Zhang, Zhen-Zhen Huang, Yi Lin, Yan Huang and Wei-Dong Zheng 172true METHODS: ARPE-19 cells were cultured in vitro, stimulated by 100nM BK for 24h. Cell morphology changes were observed by microscope, and BK receptor localization was detected through cell immunofluorescence. Changes of Ca²⁺ in BK and BR antagonist stimuli were detected by laser scanning confocal microscopy. The expressions of COX-1, COX-2, eNOS and iNOS protein in control group and BK group were detected by Western Blot.

RESULTS: After the stimulation of BK, there was no significant changes of ARPE-19 cells in morphology. Kinin B1 receptors(B1R)and B2 receptors(B2R)could be detected in ARPE-19 cells. Compared with control group, Ca²⁺ concentrations significantly increased in BK group; in B1R antagonist group and B2R antagonist group Ca²⁺ concentrations increased less than BK group; B1R and B2R antagonist group showed no obvious changes in Ca²⁺ concentrations. Compared with control group, COX-2 and iNOS protein concentrations were significantly increased in BK group(P<0.001).

CONCLUSION: BK induces the increasing expression of COX-2 and iNOS in the cultured ARPE cells through binding with either B1R or B2R.]]> 2016/7/26 10:36:27 Wen-Ting Cai, Cheng-Da Ren, Qing-Yu Liu, Qing-Quan Wei, Ya-Ru Du, Qian-Yi Wang, Jun-Ling Liu, Meng-Mei He and Jing Yu 171true METHODS: The rats were randomly divided into light exposure group and control group. Rats in light exposure group were exposed in white light(10 000lux,12h on-off, continuing 1-14d). Rats in control group were only exposed in natural light. The eyes of the rats in the two groups were removed when the rats in light exposure group acceptted intense light after 1, 3, 7 and 14d. We observed the change of retinal structure using hematoxylin-eosin(HE)staining, and observed the change of retinal ultrastructure using electron microscope. We quantified the change of retinal vascular permeability using laser scanning confocal fluorescence microscope and spectrophotometry after perfusion of Evans-blue, to evaluate the change of blood-retinal barrier.

RESULTS: At 1d after intense light exposure, the retinal ultrastructure of rats changed, such as denaturation of photoreceptor cells and falling of membranous disc outer segment and thinning of the outer nuclear layer thickness, and so on; and the longer the rats exposure to intense light, the more serious change of the retinal ultrastructure were found. At 3d later, photoreceptor cells began apoptosis. At 14d later, the outer nuclear layer became thinner obviously, and the number of cells reduce obviously. At 1d after intense light exposure, EB leaked from the retinal vascular, and at 14d later the leaking of EB was more obvious.

CONCLUSION: The photoreceptor cell of the outer nuclear layer of retina will degenerate and apoptosis, and the outer nuclear layer will be thinner, and the structure and function of blood-retinal barrier will be destroied, if the eyes of rats exposed in intense light.]]> 2016/7/26 10:36:27 Xiao-Ting Wang, Guo-Xing Xu, Wei Xu and Mao-Song Xie 170true METHODS:Allogeneic penetrating corneal transplantation in rat model was performed. SD rats were randomly divided into positive control group, GFP-DC group, 8-DC and IL-10-GFP-DC group. At 3d before keratoplasty, the rats were given tail intravenous injection with the same amount of PBS, bone marrow 8-DC(DC had cultured for 8d)from donor Wistar rats, GFP-DC after 48h transfection and IL-10-GFP-DC. Rats were observed under slit-lamp for corneal graft cases every day, and recorded rejection index and corneal graft survival time. At 14d after keratoplasty, pathologic and immunohistochemical examinations were performed.

RESULTS:Compared with GFP-DC group and 8-DC group, corneal graft survival time of IL-10-GFP-DC group was significantly longer(P<0.01); at 14d after keratoplasty, corneal opacity, edema, neovascularization and rejection index of IL-10-GFP-DC group were significantly lower(P<0.01). Pathological examination showed that in the three experimental groups corneal inflammation was lighter than the positive control group without significant central graft neovascularization. Immunohistochemistry showed: compared to the positive control group, GFP-DC group and 8-DC group, CD4⁺, CD8⁺, CD25⁺, IL-2⁺, NK⁺ and NF-κB⁺ positive cells in IL-10-GFP-DC group were lower(P<0.01).

CONCLUSION: After donor-derived immature dendritic cells pretreated, corneal graft survival was significantly prolonged, successfully induced corneal transplantation tolerance. CD4⁺, CD8⁺, CD25⁺, IL-2⁺, NK⁺ and NF-κB⁺ positive cells are involved in corneal allograft rejection regulation, IL-10-GFP-DC may reduce CD4⁺, CD8⁺, CD25⁺, IL-2⁺, NK⁺ and NF-κB⁺ positive cell infiltration, inhibit corneal transplant rejection.]]> 2016/7/26 10:36:27 Jia Li, Xue Li, Jian-Hua Sun and Bing Li 169true METHODS: Male Wistar rats were randomized into 4 groups, normal control group, diabetic control group and PGMS in group, the PGMS in groups included the doses of 50mg/kg and 100mg/kg. 1% streptozotocin(STZ)of 60 mg/kg was intraperitoneally injected in rats to establish the diabetic models. The PGMS with the doses of 50mg/kg and 100mg/kg were used to gavage in different groups of models for 12wk. Twelve weeks later, the animals were sacrificed and retinas were isolated. The aqueous humors and serums were taken, expressions of TNF-α and IL-1β protein in retinas, aqueous humors and serums were detected by enzyme-linked immunosorbent assay(ELISA), respectively. The location and the expression of TNF-α and IL-1β protein in retina tissue was detected by immunohistochemistry.

RESULTS: Twelve weeks after the use of PGMS, the level of blood glucose was not changed. ELISA showed that the expression of TNF-α and IL-1β protein in serum and retina was significantly increased in diabetic control group than in normal control group(P<0.05), but in the groups which PGMS was given reduced, lower than those in diabetes mellitus(DM)group, especially as the concentration of PGMS increased(P<0.05). But the levels of aqueous humor's TNF-α and IL-1β proteins in PGMS group were not reduced. Immunohistochemistry showed that the TNF-α protein was almost not expressed in normal control group. But the TNF-α protein was highly expressed in diabetic control group. The expression mainly located in the ganglion cell layer, the inner plexiform layer, outer plexiform layer and pigment epithelium. The TNF-α protein was weakly expressed at the group of 50mg/kg PGMS, the TNF-α protein was almost not expressed at the group of 100mg/kg PGMS. When the normal control group was detected, the IL-1β protein was weakly expressed in the outer plexiform layer. But the IL-1β protein was also highly expressed in diabetic control group.The expression mainly located in the inner plexiform layer, outer plexiform layer and pigment epithelium. The IL-1β protein was weakly expressed at the group of 50mg/kg and 100mg/kg PGMS.

CONCLUSION: PGMS can treat the diabetic retinopathy by downregulating the expressions of TNF-α and IL-1β in early diabetic retinopathy. PGMS maybe have a good control effect on early diabetic retinopathy.]]> 2016/7/26 10:36:27 Wei-Yan Zhou, Hong-Ya Wang, Xiu-Juan Du and Wei-Hong Dong 168true in vitro.

METHODS: Different concentrations(0, 0.625, 1.25, 2.5, 5.0mg/mL)of bevacizumab were exposed to ARPE-19 cells, then cell viability was analyzed by CCK-8, cell cycle was determined by flow cytometry, and the expression of E-Cadherin and fibornectin was detected by Western blot and RT-PCR.

RESULTS: The concentration as 2.5mg/mL or 5.0mg/mL of bevacizumab was shown to effectively suppress the proliferation and cell cycle of ARPE-19 cell(P<0.05). In addition, 2.5mg/mL or 5.0mg/mL of bevacizumab could downregulate the expression of E-cadherin and promote the transcription of fibronection gene(P<0.05).

CONCLUSION: High concentration of bevacizumab was able to inhibit ARPE-19 proliferation, downregulate E-Cadherin expression and promote fibronectin expression, indicating epithelial-mesenchymal transition induced by bevacizumab in ARPE-19 cell.]]> 2016/7/26 10:36:27 Chan Li and Wei Ren 167true in vivo the effect of androgen on mice tear film stability and Mucins expressions in corneal epithelial cells in BALB/c mice after orchectomy.

METHODS: With orchiectomy operation, we set up mice model. And serum androgen concentration of mice was detected by radioimmunoassay. Break-up time(BUT)of tear film was tested in the different experimental points. Mice corneal epithelia were peeled and MUC1 and MUC4 mRNA and protein levels were measured by RT-PCR and Western blot.

RESULTS: The serum androgen concentration reduced to 0ng/μL at 1wk after orchiectomy. The BUT values were 68.33±12.86s, 62.47±3.75s, 58.67±10.03s, 47.17±7.64s, 39.51±3.39s, 32.67±3.88s and 31.00±2.36s in the normal control group, sham group and in orchectomy group at 1, 2, 4, 6 and 8wk, respectively, and the BUTs were significantly shorter in the orchectomy at 2, 4, 6 and 8wk groups than those in the normal control group(all at P<0.05). MUC1 and MUC4 mRNA and proteins levels decreased with androgen level lowering(P<0.05). Mucin1 level was the lowest at 2wk after orchiectomy, and the lowest Mucin4 level was found at 1wk after orchiectomy.

CONCLUSION:In vivo, androgen regulates Mucins expressions in mice corneal epithelial cells, makes BUT shorter,and influence the stability of tear film.]]> 2016/6/29 17:05:27 Li Li, Xuan Zheng, Shuang-Mei Wang, Ge Gao and Qian-Yan Kang 166true METHODS: Totally 26 rabbits(26 right eyes)with dry eye model were studied and divided into two groups: group A(control group with PBS eye drops, n=13)and group B(amniotic extraction group, n=13). Another two rabbits were chosen as normal control.The Schirmer Ⅰ tests(SⅠt)and corneal fluorescein staining(FL)were made, and the tear total protein content, amylase activity, lactoferrin, lysozyme contents, goblet cell density were performed in two groups before treatment and 1, 2, 4 and 8 wk after treatment.

RESULTS: There were significant differences in SIT, FL scores, lysozyme activity and goblet cell density among different groups at different time points(P<0.05). But, there was no significant differences in SⅠt, FL scores, lysozyme activity and goblet cell density between two groups before treatment(P>0.05). After 8wks' treatment with PBS, the mean differences of the group A showed great changes in SⅠt, lysozyme and goblet cell density compared with those before treatment(P<0.05); but there was no significant differences in FL scores compared with those before treatment(P>0.05). As for group B, 8wks after treatment, there were statistical changes in SⅠt, FL, lysozyme(P<0.05); but there was no significant differences in goblet cell density compared with those before treatment(P>0.05). It was evident that statistical differences were observed in SⅠt, FL scores, lysozyme activity and goblet cell density between two groups at each time point(P<0.05). However, there were no significant differences in total protein, lactoferrin, amylase activity at different time points(P>0.05). Meanwhile there was no significant differences in total protein, lactoferrin, amylase activity between two groups before treatment(P>0.05). But there were significant differences in total protein, lactoferrin, amylase activity between two groups after 4 and 8 wks' treatment(P<0.05).

CONCLUSION: Amniotic extraction has significant therapeutic effect on the dry eye in rabbit model.]]> 2016/6/29 17:05:28 Juan Du, Zhi-Hui Li, Fen-Tu Zhao, Yi Shao, Nan Jiang, Xue-Fu Tang and Min-Ting Feng 165true METHODS: Cultured RF/6A cells were divided into normal control group, high glucose group and high glucose(HG)+ different concentration cordycepin groups(HG+10μg/mL group, HG+50μg/mL group, HG+100μg/mL group). The cell proliferation was assessed using cholecystokinin octapeptide dye after treated for 48h. The cell migration was investigated by a Transwell assay. The tube formation was measured on Matrigel. Furthermore, the impact of cordycepin on high glucose-induced activation of VEGF and VEGF receptor 2(VEGFR-2)was tested by Western blot analysis.

RESULTS: Compared with normal control group, cell viability markedly increased in high glucose group(P<0.05). Cordycepin inhibited RF/6A cell proliferation in a dose-dependent fashion: 10.2±0.9%, 23.4±1.5% and 31.1±1.2% inhibition as the concentrations of cordycepin were 10, 50 and 100μg/mL, respectively. The difference had statistically significant(P<0.05)compared with high glucose group. The number of cell migration were 55.6±2.70, 87.4±2.40, 65.4±2.7, 57.8±2.38, 62.4±2.77 in normal control group, high glucose group and HG+10μg/mL group, HG+50μg/mL group, HG+100μg/mL group respectively. Migration of RF/6A conspicuously increased in high glucose group(P<0.05)compared with normal control group; while showing a gradually reducing trend with the increase of cordycepin dose and a statistically significant difference compared with high glucose group(P<0.05). The number of tube formation were 18.7±2.08, 25.7±1.52, 19.9±1.57, 16.3± 2.51, 5.67±1.72 in the abovementioned group. Similarly showing a gradually reducing trend with the increase of cordycepin dose and a statistically significant difference with high glucose group(P<0.05). In addition, the number of tube formation of RF/6A in high glucose group significant increased compared with normal control group(P<0.05). The expression of VEGF and VEGFR-2 dramaticlly increased in high glucose group vs normal control group, oppositely gradually decreased with the increase of cordycepin concentrations, and had a statistically significant difference vs high glucose group(P<0.05).

CONCLUSION: Cordycepin can suppress the proliferation, migration and tubu formation of RF/6A in high glucose condition, might via inhibiting expression of VEGF and VEGFR-2.]]> 2016/6/29 17:05:28 *, Xiao-Feng Sun^* and Ming-Ying Lai]]> Xiao-Li Zhu^*, Xiao-Feng Sun^* and Ming-Ying Lai 164true METHODS: The HLE cell oxidative damage model induced by high concentration glucose was established, and was intervented with different concentrations of EGCG. Cell viability was determined by MTT assay, cell morphology was investigated by convert microscope, cells apoptosis was assayed by flow cytometry with Hoechst-PI staining. Moreover, the levels of super oxide dismutase(SOD), glutathione peroxidase(GSH-Px)and malondialdehyde(MDA)in supernatant were also tested after different treatment either with high concentration glucose or with different concentrations of EGCG.

RESULTS: MTT results showed that HLE cells activity increased to 50.33%±3.52% and 63.33%±4.63% after treated with 10 μmol/L and 100 μmol/L EGCG respectively, the difference was statistically significant compared with oxidative injury group(32.67%±3.10%)(P<0.05); HLE cells maintained better morphology intervented with EGCG under high glucose conditions, the number of apoptotic cells reduced, SOD and GSH-Px level within HLE cells increased and MDA levels decreased.

CONCLUSION: EGCG plays its strong antioxidant effect by increasing SOD, GSH-Px content and decreasing MDA content in cells, therefore provides a reliable experimental basis for the search for effective prevention and treatment of cataract drug.]]> 2016/5/31 16:15:53 Ting Chen,Ping Liu,Jia-Xiang Wang,Duo Shan,Wen Che and Li-Juan Zhang 163true METHODS: Twenty-five Balb/c mice were divided into five groups at random: A: model group, B: artemisinin topical treatment group, C: oral artemisinin group, D: dexamethasone topical treatment group, E: negative control group. Balb/c mice were first sensitized with mixture of ragweed crude extract and complete Freund's adjuvant by left footpad and root of tails injection at days 0. At days 7 and 14, mice were strengthen sensitized by intraperitoneal injection. Negative control group replacd with a mixture of equal amounts of Freund's adjuvant and PBS mixture. Treatment time was between 21-27d: group B received 1% topical artemisinin eye drops 4 times per day; group C received artemisinin at doses of 600mg/kg orally once per day; group D received topical dexamethasone eye drops 4 times per day. The model and naive groups replaced with solvent for control. After treatment, mice were excited by ragweed pollen crude infusion drops. Within 1h after the excitation, the eyes were taken for histopathology testing, the serum for detection of specific IgE for ragweed pollen.

RESULTS: After excitation, allergy symptoms of model group were more significantly compared with negative control group and the treatment groups; allergy symptom score, mast cell degranulation ratios and the ragweed pollen specific IgE in serum in the topical treatment groups and oral group were higher than that in the negative control group, but significantly lower than that in model group.

CONCLUSION: Artemisinin topical treatment and oral treatment for mice could inhibit the symptoms of pollen allergic conjunctivitis, early-phase response of mast cell degranulation ratio and specific IgE generation for ragweed pollen, suggesting that artemisinin has some therapeutic effect for pollen allergic conjunctivitis.]]> 2016/5/31 16:15:53 Jing-Jing Zhu and Bing Li 162true METHODS:Human lens epithelial cell line(HLE-B3)were cultured in normal condition or with H₂O₂ for 24h. Total RNA were extracted for gene expression profiling assay and gene ontology analysis was performed for the significantly up-regulated genes using bioinformational database DAVID. The elevated expressions of up-regulated genes in HLEC after oxidative stimulation were confirmed by RT-qPCR. The apoptosis of HLEC induced by oxidative damage was detected using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide(MTT)assay and flow cytometry.

RESULTS:Gene expression profiling assay demonstrated that 367 genes were up-regulated in HLEC after oxidative stimulation. These genes were enriched in 310 biological processes mainly associated with p53-related signaling pathways, apoptosis, programed cell death and etc. Six genes mainly pro- or anti-apoptotic, including BCL2A1,TP53I3,FAS,ZMAT3,DDB2 and BCL2L1, were confirmed to be up-regulated by oxidative stimulation using RT-qPCR(P<0.05). Results of MTT assay and flow cytometry showed that apoptosis of HLEC gradually appeared after cells were treated with H₂O₂ and became the main consequence of oxidative stimulation.

CONCLUSION:Oxidative stimulation can induce up-regulation of proapoptotic genes and lead to apoptosis of HLEC, even though antiapoptotic genes can also be promoted.]]> 2016/5/3 15:55:03 Lin Mei,Song Wang,Bo Yan,An-Gang Yang,Jing Zhao and Hong Yan 161true METHODS:HLEC was subcultured and divided into negative control group, oxidative injury group, lycopene low dose group and lycopene high dose group. Cell viability was assayed by MTT colorimetric. Cell morphological changes were detected by electron microscope. Reactive oxygen species(ROS)levels were detected with DCFH-DA fluorescent probe. Content of superoxide dismutase(SOD), glutathione peroxidase(GSH)and malondialdehyde(MDA)in supernatants were detected by spectrophotometer.

RESULTS:Lycopene could obviously inhibited UV-induced decline in cell activity, reduce UV-induced ROS generation within HLEC, cause SOD, GSH-Px levels increased and MDA levels decreased.

CONCLUSION:Lycopene plays its strong antioxidant role in increasing the intracellular SOD and GSH-Px content levels and decreasing MDA levels, which provide reliable experimental basis for prevent and treatment of cataracts.]]> 2016/5/3 15:55:03 Jing-Wen Sun and Jing Hao 160true METHODS:Sixty SD rats were randomly divided into diabetic group and control group. Diabetic rat model was produced by intraperitioneal injection of 1% STZ with 60mg/Kg weight. The rats in control group received intraperitioneal injection of normal saline with same dosage. After injection, the rats were sacrificed and eyeballs were enucleated for HE staining, the retina fluorescence angiography, TUNEL and Western Blot detection at 1, 2 and 4mo for the expressions of HMGB1 and NF-κB.

RESULTS:Compared with the control group, the retinal cells disorder, cell densities decreases, microvasculars occlusion were founded with inner and outer nuclear layer thinning and ganglion cell apoptosis. The fluorescence angiography showed that peripheral capillaries became circuitous and vascular occlusion and non-perfusion area could be seen. The expressions of HMGB1 and NF-κB were higher than those of control with time dependence and they had significant positive correlations(P<0.05).

CONCLUSION:The expression of HMGB1 increases in diabetic rat retina, which may involve in the occurrence of diabetic retinopathy through the NF- κB pathway.]]> 2016/5/3 15:55:04 Shuang Jiang and Hai-Yue Xu 159true METHODS: Sixty 3-week-old C57BL/6 mice were randomly divided into experimental groups and normal groups(NC)which totally include four groups: form-deprived 3wk group(FDM3W, n=20), form-deprived 4wk group(FDM4W, n=20), FDM3W normal control group(FDM3W-NC, n=10)and FDM4W normal control group(FDM4W-NC, n=10). Mice in experimental groups were treated with diffuser wearing on right eyes(MD-T), and their left eyes were naturally exposed as their self-control group(MD-C). The normal control groups were free of all the treatments, but the same measurements were performed at the same time-point. Refractive status was examined at 3 and 4wk after treatments in all mice. The histological analysis was applied to assess the changes of the sclera and the retina. The immunohistochemical staining and quantitative real-time PCR(QRT-PCR)were applied to investigate the expression of Smad1 protein and mRNA in retina.

RESULTS:(1)There was no significant difference between right and left eyes in every group before experiment. The normal control eyes showed physiological farsighted development and the MD eyes demonstrated relatively myopization. After experiments, a significant myopia shift had been induced in MD-T compared with MD-C and NC(P<0.05).(2)Histopathologic examination showed posterior sclera and retina in MD-T were thinner than in MD-C and NC groups. Compared with MD-C and NC groups, the expression of Smad1 in retina in MD-T was significantly decreased(P<0.01).

CONCLUSION:The expression of Smad1 in retina of form-deprived myopia was down-regulated, and Smad1 likely to be involved in the development of myopia through the transduction of retinal signal.]]> 2016/3/28 15:32:19 Qian Cui,Xian Yang,Cheng-Ye Che,Gui-Bo Liu and Qing Wang 158true 7 on hypoxia-induced apoptosis of retinal ganglion cells in mice.

METHODS: According to the different factors, retinal ganglion cells of the mouse were randomly divided into four groups: control group(G1), hypoxia group(G2), hypoxia + agonist(BzATP)group(G3), hypoxia + antagonistic agent(BBG)group(G4). Methyl thiazolyl tetrazolium(MTT)method was used to detect the survival rate of cells; the rate of cell apoptosis was determined by Annxin V/PI staining flow cytometry; Western Blot analysis was employed to detect the protein expressrion levels of cleave-caspase-3 and cleave-PARP.

RESULTS: Compared with control group, the results significantly indicated the survival rate of cells decreased and the rate of apoptosis increased treated with hypoxia. In addition, the protein levels of cleave-caspase-3 and cleave-PARP were remarkably higher than the control group. BzATP markedly augmented the apoptosis induced by hypoxia, and BBG pretreatment observably decreased the hypoxia-induced apoptosis.

CONCLUSION: P2X₇ purinoceptor could be activated by hypoxia, and participate in the apoptosis of retinal ganglion cells.]]> 2016/3/28 15:32:19 Xu-Hong Hao,Yong Pei and Xiao-Nan Sun 157true METHODS:Blood samples from 10 cases of VKH patients and 10 cases of normal control were collected to extracted total RNA in plasma. The cDNA was synthesized by reverse transcription, and then underwent real time PCR in a 96 well plate from miRCURY LNA^TM Universal RT microRNA PCR panels.Original Ct value was obtained. The Exiqon GenEx qPCR analysis software was used to analyse the original data and the differential miRNAs was obtained and analyzed by bioinformatics.

RESULTS:The panels contained 179 miRNAs classes from human individuals. In comparison with the healthy controls,there were 20 differential miRNAs, in which 12 miRNAs were up regulated and 8 miRNA were down regulated(all P<0.05). These microRNAs participated in the regulation of various signaling pathways.

CONCLUSION:The VKH patients and healthy controls have significantly different expression profiles of microRNA.The differential expression of microRNA may be involved in the pathogenesis of VKH.]]> 2016/3/28 15:32:19 Xin-Qiao Zhang and Hong Wang 156true METHODS: A total of 64 C57BL/6 mice which were 3～4wk were randomized into FDM-21d group(n=16),a normal control group(n=16); FDM-28d group(n=16)and a normal control group(n=16). FDM models were established by sutured 0.5mL PCR plastic caps to the skin surrounding the right eye for 21 days and 28 days in experimental group, and the fellow eyes served as the self-control eyes. Diopter and axial length of all eyes was tested by retinoscopy optometry and vernier calipers before and after the form deprivation. As the models established, the sclera of the mice were obtained and hematoxylin and eosin(H-E)staining were used for observing the morphological change in the tissue. Immunohistochemical staining and fluorescence quantificational real-time polymerase chain reaction(QRT-PCR)were applied to investigate the expression of BMP-2 protein and mRNA.

RESULTS: After 21 and 28d,the diopter of the deprived eyes was -1.60±1.03D and -3.10±1.19D and the axial length was prolonged by 16±12μm and 21±13μm respectively. The differences were statistically significant, compared to self-control eyes and to normal control groups(P<0.05). After stained by H-E, the sclera from the deprived eyes became thinner and the sequence of collagenous fiber disappeared. The results from the immunohistochemical staining and QRT-PCR showed that mRNA and protein of BMP-2 decreased significantly, and the differences were statistically significant, compared to self-control eyes and to normal control groups(P<0.05).

CONCLUSION: The expressions of the BMP-2 in sclera down-regulate significantly in FDM eyes,which suggests that BMP-2 may play an important role in sclera remodeling during myopia development.]]> 2016/3/2 8:19:50 Yu Zhang,Xian Yang,Li-Ping Jiang,Gui-Bo Liu and Shuang-Shuang Wang 155true METHODS: One hundred male SD rats were randomly divided into 2 groups: diabetic model group(80 rats induced by STZ)and blank control group(20 rats). Then the diabetic model group was randomly divided into 4 subgroups. The group DM(diabetic group)had 20 rats with normal diet and 2mL physiological saline for daily gavage. PSP gavaged groups included three subgroups and with 20 rats in each group, group L(low dose of PSP gavage group, 200mg/kg), group M(medium dose of PSP gavage group, 400mg/kg), group H(high dose of PSP gavage group, 800mg/kg). Every group was gavaged with 2mL PSP. Twenty rats in group BC(blank control group), and they were gavaged saline of equal dose as control. After the success of modeling, 5 groups were under the same conditions of feeding, and gavaged every day. At 2, 4, 6, 8, 10 and 12wk, anterior segment examination and FFA were given. At 4, 8 and 12wk, F-VEP and ERG were given. Fasting blood glucose(by glucose oxidase method)was measured through inner canthus vein blood.

RESULTS: Compared to BC group, fasting blood glucose levels were significantly higher in group DM, L, M and H(P<0.05). Compared to DM group, fasting blood glucose level decreased in L, M, H group(P<0.05). The fasting blood glucose levels of L, M and H groups showed a time and dose dependent relationship. After anterior segment examination, there were 6 rates in DM group, 3 in L group, 1 in M group with different degrees of cataract symptoms which became more serious with time. The BC group was not found any abnormal in the anterior segment. Examined by F-VEP, the rats in group DM showed extension of the P100 peak latency. Compared to the group DM, PSP gavaged group showed a shortened of the extension of the P100 peak latency, while the group BC had no obvious change. In the ERG examination, the rats in group DM showed that amplitude of Max-R a, b wave decreased by 51.2%, 59.8% and amplitude of Cone-R a, b wave decreased by 31.4%, 41.2%. The Ops OS value and amplitude of 30Hz Flicker N1-P1 decreased, there was a significant difference compared to group BC. PSP gavaged group was better than group DM. In the FFA examination, in group DM, we could find the typical manifestations of diabetic retinopathy background fluorescence enhancement, distortion of the blood vessels and blood capillary dilate, retinal vascular leakage of fluorescein and intraretinal hemmorhages compared with the group BC. PSP gavaged group was better than group DM.

CONCLUSION: PSP can effectively reduce the blood glucose levels of diabetic rats,and also can delay the process of ocular complications in diabetic rats, which have an obvious therapeutic effect on ocular lesions. The mechanism is likely to improve the metabolism of glucose and lipid metabolism, increase the amount of glucose tolerance, and play a protective role in diabetic lens metabolism and retinal microvascular disease.]]> 2016/3/2 8:19:50 Yi Wang,Guo-Qing Peng,Di Chen,Chao Han,Chun-Run Miao,Chao Su,Bao-Jin Lu and Shan-Long Feng 154true METHODS: Sixty BALB/c mice were divided into three groups(experimental group A, experimental group B and control group C), 20 mice in every group and their corneas in the right eyes were burned with alkali(1mol/L NaOH). The experimental group A received recombinant canstatin proteins drops with 3μg/mL. The experimental group B received recombinant canstatin proteins drops with 5μg/mL and the control group C was treated with physiologic saline. At different time points(1, 3, 7 and 14 d)after alkali burns, the mice were killed and the growth of epithelial defect and corneal neovascularization(CNV)were observed with an operation microscope. The expressions of MMP-2 and TIMP-2 in cornea were measured by the Western blot technique, and the results were analyzed by enhanced chemiluminescent(ECL).

RESULTS: The areas of epithelial defect and corneal neovasularization significantly reduced in mice treated with recombinant canstatin proteins compared to mice treated with physiologic saline at 3, 7 and 14d after alkali-induced injury( all P<0.01); the neovasularization was suppressed and the area of CNV was less than that in control group C(all P<0.01). Western blot analysis showed that the expression levels of MMP-2 in experimental group A and B were significantly lower than that in control group C(P<0.01)and the expressions of TIMP-2 in experimental group A and B were significantly higher(P<0.01); the level of MMP-2 in experimental group B were lower than that in experimental group A on day 14(P<0.05), while the level of TIMP-2 in experimental group B were significantly higher than that in experimental group A on day 7 and day 14(P<0.05).

CONCLUSION: Recombinant canstatin proteins may suppress the expression of MMP-2, upregulate the expression of TIMP-2 in cornea cells and the infiltrated inflammatory cells, lower the rapid resolution of cornea and ulceration, and play a vital role in the remodeling of alkali treated cornea in mice.]]> 2016/2/3 8:48:35 Ming Lu and Jing Zhu 153true METHODS:Different concentrations of PDGFR-α shRNA were blended with lip2000,and the final scale of PDGFR-α shRNA/lip2 000 complex was 1:1, 1:2 and 1:3 respectively(the concentration of PDGFR-α shRNA was respectively 2μg, 3μg and 4μg).All the complexes were cultivated in human retinal pigment epithelium(HRPE)for 24h after transfection and then respectively injected 0.1mL to rabbit vitreous cavity.Forty healthy adult rabbits were seleceted in this study and randomly divided into colored balanced salt solution(BSS)group(N), comprising lipofectamine^TM2 000 HRPE cell dilution group(group A). The highest transduction efficiency of 1.0, 1.5 and 2.0μmol/L containing PDGFR-α receptors shRNA, lipofectamine^TM2 000 of HRPE cell dilution were selected(respectively group B, C and D)with 8 eyes each, the right eyes as the experimental eye. The extent of PVR was observed by indirect ophthalmoscope; and slice staining situation was observed by immunohistochemistry.The fundus changes were observed by histopathology.

RESULTS:The highest transduction efficiency of PDGFR-αshRNA/lip2 000 ratio was 1:2.The extent of PVR, the histopathology changes and the immunohistochemistry of PDGFR-α in group B,C and group D were significantly lower than that in group A, while group D was much lower than those of group B and C.

CONCLUSION:PDGFR-αRNA interference could inhibit the formation of experimental PVR.]]> 2015/12/28 16:15:58 Zhu Meng,Yan-Yi Peng and Xian-Jie Qin 152true METHODS: Cultured 3～5 passage RPE cells of C57BL/6 mouse were incubated with rod outer segments(ROS)suspension(containing ROS 1×10⁷ /ml)at 37℃, then cells were rinsed at different times(30,60,120,180,240min)to terminate the phagocytosis. The kinetics of phagocytosis was measured by double-fluorescent labeling. The activity levels of MERTK, Ras, MEK and MLCK at different incubation times were measured by Western Blot with antibodies of MERTK, Ras and phospho-antibodies of MEK and MLCK, respectively. To repeat the measurement of the phagpcytic kinetics and activity levels of MERTK, Ras, MEK and MLCK at different incubation times after interference to Ras and Mertk gene in RPE cells by plasmid transfection.

RESULTS: The phagocytic kinetics showed that the ingestion occurred at 30min of incubation. Ingested ROS by RPE cells increased until saturated at 180min. The protein levels of MERTK, Ras, MEK and MLCK in RPE cells increased during all the incubation periods compared with control group(P<0.05). After interference to Ras and MERTk gene in RPE cells by plasmid transfection, the protein levels of MERTK, Ras, MEK and MLCK and the quality of ingested ROS sustained at the lower levels during all the incubation periods, only a few ingested ROS were seen at 180 min. Compared with untransfected RPE cells, the protein levels of MERTK, Ras, MEK and MLCK and the quality of ingested ROS in siRas-RPE cells and siMERTK-RPE cells decreased distinctly at 120 min and 180 min during incubation(P<0.05).

CONCLUSION: Ras-MEK-MLCK-myosin signal pathway is the downstream of MERTK receptor in the phagocytic process of RPE cells from mice.]]> 2015/12/28 16:15:58 Yu-Zhao Sun,Ruo-Shuang Zhang and Feng Gu 151true reinforcing yang from yin of a traditional Chinese patent medicine called YouguiWan for PI3K/Akt pro-survival signal channel in diabetic retinopathy rat.

METHODS: A total of 80 SPF SD rats were selected, 10 rats were selected to set up normal group. The remaining 70 rats were given streptozotocin(60mg/kg)peritoneal injection combined with intravitreal injection of VEGF(0.05μg)to establish proliferative diabetic retinopathy rat model. After the model successful, the rats were divided into four groups: YouguiWan high dose group, YouguiWan middle dose group, YouguiWan low dose group and model group. The treatment groups were fed daily YouguiWan concentrate, model group and the normal group received daily doses of distilled water. After intragastric administration for 3mo, the PI3K, Akt expression of retinal tissue were observed by SP-immunohistochemistry and Western-blot method respectively.

RESULTS: Immunohistochemical observation showed PI3K, Akt expression immunohistochemical staining, were brown particles on the retina. Normal PI3K, Akt expression weakly positive immunological reaction unit located in the ganglion cell layer. The kernel was also a small amount of expression. Model group, each YouguiWan treatment group rat retinal ganglion cell layer, inner plexiform layer and the inner and outer layer of the particles had a positive expression. Compared with the model group, YouguiWan high dose group PI3K, Akt expression were decreased. YouguiWan low dose group decreased expression was not obvious. Compared with the low dose group, YouguiWan high dose group PI3K, Akt expression was decreased. The expression of the strength of the YouguiWan, high dose group showed no significant difference. Western-blot test results showed that compared with the normal group, model group, treatment group, the expression difference was statistically significant(P<0.01). Model group, treatment group, PI3K, Akt protein levels were significantly increased. Compared with the model group, the YouguiWan high-dose group was statistically significant(P<0.05). YouguiWan high dose group PI3K, Akt protein expression were decreased, YouguiWan low dose group was no significant difference(P>0.05). Compared with the low dose group, YouguiWan high-dose group was statistically significant(P<0.05). YouguiWan high dose group PI3K, Akt expression levels were decreased. YouguiWan high dose group was not statistically significant difference(P>0.05).

CONCLUSION: Reinforcing yang from yin of YouguiWan can affect PI3K, Akt protein expression and inhibit PI3K/Akt signaling pathway, delay the disease process of diabetic retinopathy, propose new ideas and scientific basis for the treatment of diabetic retinopathy and blindness prevention treatment.]]> 2016/11/23 14:07:30 Huan Li, Xiang-Xia Luo, Yu-Pei Feng and Han Wang 150true METHODS: Neonatal C57BL/6J mice were divided randomly into three groups: normoxic control group, PBS injection of OIR group and VAS2870 injection of OIR group. The mice of the latter two groups were exposed to 75% oxygen from the postnatal 7d(P7)to the postnatal 12d(P12)to induced OIR. VAS2870 was administered by intravitreal injection(0.5μL)in a mice model of OIR in P12. Another set of mice model of OIR were received a similar treatment with PBS. All eyes were collected at P17. The right eyes were whole mounted and stained with Lectin to observe the growth of retinal vessels; The eyes were enucleated to assess the levels of reactive oxygen species ROS/RNS. The expression of vascular endothelial growth factor(VEGF)and Nox4 mRNA were detected by western blot and RT-PCR, respectively.

RESULTS: In the retina of OIR, VAS2870 reduced the retinal avascular area and neovascularization, the hypoxia-induced increase in ROS levels, the protein expression of VEGF and gene expression of Nox4 mRNA.

CONCLUSION: NOX4 enzyme inhibition with VAS2870 has potent anti-oxidative stress effects in the retina, indicating its potential as a treatment for retinopathy of prematurity.]]> 2016/11/23 14:07:30 Yan-Fang Hao, Qiao-Ping Zhu and An-Ming Xie 149true METHODS:A total of 18 aged 3-week kittens were randomly divided into monocular deprivation, strabismus and normal groups. All types of amblyopia were developed in the experimental eyes that were detected by P-VEP 12wk later. The cats were killed and the immunocytochemistry staining method were applied to observe under the light microscope the changes of distribution and positive cells areas of NO and GABA across the amblyopic retinal, compared to that from the normal cats of identical age.

RESULTS: The P-VEP showed that the amplitude of wave P1 was lower(P<0.05)and the P1 latent time was longer(P<0.05)in two types of amblyopic cats than those in the normal cats. Compared to the normal cats, the NO and GABA positive cells areas were obviously reduced(P<0.05)across the retina in the amblyopic cats. But no significant difference was found between two kinds of amblyopic cats.

CONCLUSION:The NO and GABA play an important role in the formation of amblyopia in the level of retinal.]]> 2016/10/25 14:35:36 Han-Min Wang, Ao Rong, Li-Juan Mo, Qing-Song Li and Xing-Ru Zhang 148true 2O₂.

METHODS: RPE cells were subculture, and they were divided into negative control group: cultured with normal culture medium; oxidative injury group: 100 μmol/L H₂O₂ treated for 12h; quercetin low dose group: 100 μmol/L quercetin incubated for 24h then treated with 100 μmol/L H₂O₂ for 12h; and quercetin high dose group: 100 μmol/L quercetin incubated for 24h then treated with 100 μmol/L H₂O₂ for 12h. Cell viability were tested by MTT colorimetric detection, apoptosis rate was detected by flow cytometry, apoptotic cell morphology was observed by Hochest33258 staining, expression of catalase(CAT), superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)were tested by colorimetric detection.

RESULTS: Quercetin inhibited H₂O₂-induced cell viability decreased in RPE cells, after treated with different concentrations of quercetin, RPE cells activity increased to(79.67±4.98)% and(83.00±3.60)%, which had statistical significance difference compared with oxidative damage group(48.93±3.39)%(P<0.05). After treated with different concentrations of quercetin, the apoptosis rate of RPE cells decreased to(23.23±3.29)% and(16.23±1.94)%, respectively, which had statistical significance difference compared with oxidative damage group(38.03±4.76)%(P<0.05). In addition, quercetin also increased the expression of CAT、SOD、GSH-Px in RPE cells, which had statistical significance difference compared with oxidative damage group.

CONCLUSION:Quercetin effectively inhibited H₂O₂-induced RPE cells damage by improving cell antioxidant enzyme activity, which provide reliable experimental basis for the treatment of injuries in RPE cells.]]> 2016/10/25 14:35:36 Jia-Jun Zhang, Xin-Xin Li, Bao-Shi Chen and Li-Juan Liu 147true METHODS: The model of alkali burn corneal was made. Different times corneas were taken to electron microscope for vascularization, and examined the expression of VEGF-C/D and VEGFR-3 in l, 3, 5, 7, 14, 28d. The other rat cornea after alkali burn were divided into four parts to penetrate keratoplasty, containing only blood vessels in the cornea(group A), angiogenesis and lymphangiogenesis(group B), lymphangiogenesis degenerating period(group C), angiogenesis degenerating period(group D). In addition, there are also normal groups(group N)to compare the RI values and survival time of corneal graft.

RESULTS: Electron microscopy showed that, when the first 7d rat cornea appeared neovascularization after alkali burn, but not lymphangiogenesis. The occurrence of new blood vessels and lymphatic in 2wk. There were no obvious lymphangiogenesis in 5wk and the angiogenesis gradually subside in 8 wk. The expression of VEGF-C/D and VEGFR-3 in the corneas of rats were up-regulated in the third days after the injury, and reached its peaks at 5d. The average survival time of group N, A, B, C, D were(14.25±0.62)d,(9.35±1.02)d,(5.06±1.13)d,(8.71±0.83)d,(9.44±1.05)d after transplant cornea. Compared to the rest of the group, group B plant average survival time significantly shortened(P<0.05), while compared with group B, the survival time of A, C, D groups were significantly longer(P<0.05).

CONCLUSION: VEGF - C/D and VEGFR-3 are expressed significantly after corneal alkali burn. New lymphatic vessels can accelerate high-risk corneal transplantation immune rejection.]]> 2016/9/19 17:26:48 Qi-Ming Wang, Xin-Yue Zhao and Zhi Wang 146true METHODS: BMSC were divided into blank control group(without transfected BMSC), negative control group(empty vector without BDNF gene transfected BMSC)and experimental group(BDNF gene transfected BMSC). The expression of BDNF mRNA in BMSC was measured by Realtime PCR, and the expression of BDNF in BMSC was measured by ELISA.

RESULTS: The BDNF mRNA expressions of 3, 4, 5, 6, 7 and 8-generation BMSC cells in the experimental group were higher than those in the blank control group and negative control group. The differences were statistically significant(P3: F=491.788, P<0.05; P4: F=380.112, P<0.05; P5: F=1854.929, P<0.05; P6: F=224.540, P<0.05; P7: F=619.155, P<0.05; P8: F=10.092, P<0.05). As the BMSC cells in the experimental group passaging, the BDNF mRNA expressions in the experimental group decreased. The difference of BDNF mRNA expression among different passage cells was statistically significant(F=298.603, P<0.05). The BDNF secretion of 3, 4, 5, 6, 7 and 8-generation BMSC cells in the experimental group were higher than those in the blank control group and negative control group. The differences were statistically significant(P3: F=520.609, P<0.05; P4: F=734.520, P<0.05; P5: F=152.847, P<0.05; P6: F=80.372, P<0.05; P7: F=96.083, P<0.05; P8: F=38.532, P<0.05). As the BMSC cells in the experimental group passaging, the BDNF secretion decreased. The difference of BDNF secretion among different passage cells was statistically significant(F=230.084, P<0.05).

CONCLUSION: Long-term expression of BDNF in BMSC can be enhanced by genetic engineering.]]> 2016/9/19 17:26:48 Mao-Song Xie, Guo-Xing Xu and Li-Bin Huang 145true METHODS: In vitro cultured human umbilical vein endothelia cells in hypoxic environment for 6h, then maintained under normal oxygen condition for 3h, 6h, 12h, 24h. miR-132 and peroxisome-proliferator-activated receptor-γ coactivator-1α(PGC-1α)expression was detected by quantitative Real-time polymerase chain reaction and Western blot analysis. Human umbilical vein endothelial cells transfected miR-132 mimic and miR-132 inhibitor(anti-miR-132)were measured by quantitative Real-time polymerase chain reaction and Western blot.

RESULTS: miR-132 and PGC-1α expression was significantly(P<0.01)upregulated in the hypoxic environment of cells at 3h compared with the normal oxygen condition. After cells transfection, the hypoxic environment the miR-132 and PGC-1α expression were markedly increased compared with the normal oxygen condition. The cells transfected miR-132-mimic, the expression of the miR-132 and PGC-1α were higher than that of transfected anti-miR-132 and contrast group(P<0.01).

CONCLUSION: miR-132 level is highly expressed in the HUVEC under hypoxia and may be an effect of regulation for PGC-1α.]]> 2016/9/19 17:26:48 Li-Xin Zhang and Li-Juan Tao 144true METHODS: Cells were divided into two groups, including blue light group and control group. The 35W white light lamp with blue filter was used to establish damaged RPE cell model in vitro. Blue ray wavelength ranged between 470nm and 520nm. And the light intensity was about 2 000Lx. After exposure to blue light, we tested the proliferation of human retinal pigment epithelial cells by CCK-8 kit. And then expression of miR-103 was measured by the real-time PCR.

RESULTS: Exposure to blue light inhibited the proliferation of human retinal pigment epithelial cells and increased the expression of miR-103. Moreover, up-regulation of miR-103 inhibited the proliferation of human retinal pigment epithelial cells, and down-regulates miR-103 promoted the proliferation of human retinal pigment epithelial cells.

CONCLUSION: Blue light inhibits the proliferation of human retinal pigment epithelial cells by the up-regulation of miR-103.]]> 2017/7/24 10:38:37 Hong-Na Zhu, Ying Qiao, An-Le Su, Ting Zhang, Zhong-Yang Sun and Hou-Cheng Liang 143true METHODS:A total of 60 male Sprague-Dawley(SD)rats were randomly divided into normal control group(NCG), model group(MG), low-dose curcumin group(LDCG)and high-dose curcumin group(HDCG)(n=15 per group). RIRI was generated by anterior chamber perfusion of normal saline to the right eye. Rats in LDCG and HDCG received an intraperitoneal injection of 20mg/kg/d and 100mg/kg/d curcumin respectively, at 30min before RIRI and once daily after RIRI. Retinal structure and inflammation were evaluated after hematoxylin and eosin-stained(HE)staining. Western-blot and enzyme-linked immunosorbent assay(ELISA)were used to measure the level of IL-23 and IL-17 expressions after RIRI.

RESULTS: The retinal structure of NCG was normal. Retinal edema, empty spaces or loosely packed cells and inflammatory cell infiltration were observed in MG and LDCG groups, whereas the morphological changes in HDCG group were improved as compared to MG and LDCG groups. Western-blot assay and ELISA showed that IL-23 and IL-17 expressions increased significantly after RIRI(vs NCG, P<0.01). Moreover, curcumin reduced IL-23 and IL-17 expressions significantly(vs MG, P<0.01).

CONCLUSION: Curcumin can inhibit leukocytes infiltration and improve the retinal pathologic changes. Furthermore, curcumin can reduce retinal IL-23 and IL-17 expressions significantly in a dose-dependent manner.]]> 2017/7/24 10:38:37 Hai-Jiang Zhang and Liang Liang 142true in situ keratomileusis(Epi-LASIK)in rabbit cornea.

METHODS: Thirty rabbit corneas were performed with Epi-LASIK. All eyes were randomly divided into three groups: eyes treated with amniotic extraction(AE group), eyes treated with 1g/L dexamethasone(hormone group)and eyes treated with solvent(solvent control group). Haze grade evaluation was performed under the slit lamp after Epi-LASIK for 1, 4 and 8wk. The repair of corneal epithelium was observed by using HE staining, and the expression of NF-kB protein P65 was detected by immunohistochemistry. The expression levels of inflammatory cytokines(TNF-α, TGF-β1 and IL-1)and anti-inflammatory cytokines(IL-4, IL-10 and IL-13)were determined by ELISA.

RESULTS: HE staining showed that the basal cells of corneal epithelium were more uniform and arranged regularly in AE groups after Epi-LASIK for 1wk as compared with the hormone group and the solvent control group. After 4wk, there were a few of new collagen fibers in the superficial stroma of AE group, forming a small amount of scar. After 8wk, the corneal stroma of AE group showed a small amount of new collagen fibers, arranged regularly, and rarely formed scar. At the early stage(1 and 4wk), AE treatment has an obviously effect on inhibiting the secretion of inflammatory factors(TNF-α, TGF-β1 and IL-1)and anti-inflammatory factors(IL-4, IL-10 and IL-13), and the difference was statistically significant(P<0.01). Moreover, the activation of the NF-kB signaling pathway was significantly inhibited by treatment with AE in the early postoperative period(1 and 4wk).

CONCLUSION: Amniotic extraction may reduce the inflammatory response in corneal epithelial cells by inhibiting the NF-kB signaling pathway, thereby inhibiting the formation of collagen and scar and the occurrence of haze.]]> 2017/7/24 10:38:37 Shi-Gang Xia, Yu-Fang Wang, Qi-Guo Xiao and Hua Peng 141true METHODS: In this cohort study, data of 19 keratoconus eyes of 18 patients which undergone ICRS implantations were gathered before and 3mo after surgery. Uncorrected visual acuity(UCVA), best corrected visual acuity(BCVA), manifest refraction, IOL power calculation formulas, axial lenght(AL)and keratometry were analayzed.

RESULTS: Mean age of participants was 29.58±0.6. UCVA improved from 0.84(0.35)logMAR to 0.43(0.31)logMAR significantly(P<0.001). BCVA and AL didn't change significantly after 3mo. All Sphere, cylinder and spherical equivalent(SE)were improved significantly(P<0.001). On the other hand, keratometry 1(K1)and keratometry 2(K2)decreased significantly. It was a considerable change in SRK/Ⅱ(P<0.001), Hoffer Q(P<0.001)and Holladay Ⅰ(P<0.001)after 3-month's follow-up. Among this formula SRK/II had the lowest change.

CONCLUSION: In addition to improvement in visual, refractive, and keratometry outcomes after Keraring implantation, there was a significantly changes in IOL calculation formulas values. However, ICRS procedure doesn't interfere considerably AL in eyes, but it seems reduced keratometric values lead to IOL power calculations more accurately and all formulas suggested same IOL power.]]> 2017/6/26 0:00:00 Khosrow Jadidi, Saman Mohazzab-Torabi, Farhad Nejat, Shiva Pirhadi and Hossein Aghamollaei 140true METHODS: The recombinant pLenti-CMV-PEDF-EYFP vector was constructed with pLenti-CMV-hChR2-EYFP and PEDF gene, and then infected into hUCMSCs to obtain the PEDF-hUCMSCs. The expression of PEDF in hUCMSCs was identified by confocal microscopy and ELISA. The Cell Counting Kit(CCK8)was used to assess the cell viability of PEDF-hUCMSCs. The influences of pLenti-CMV-PEDF-EYFP infection on the passage and phenotypes of hUCMSCs were assessed by microscope and flow cytometry.

RESULTS: The recombinant pLenti-CMV-PEDF-EYFP was successfully constructed and efficiently infected into hUCMSCs. The expression of PEDF was positively detected in the PEDF-hUCMSCs. No significant difference was observed on cell viability between PEDF-modified hUCMSCs and hUCMSCs(P>0.05). The infection of pLenti-CMV-PEDF-EYFP had no obvious influences on the proliferation, morphology and phenotypes of hUCMSCs.

CONCLUSION: PEDF was expressed effectively in the hUCMSCs modified with the recombinant lentivirus. The preliminary results from this study provide more evidences for further study of using PEDF-hUCMSCs to treat retinitis pigmentosa.]]> 2017/6/26 10:38:48 Hui Wen, Yun Zhang, Xi-Luan Ji, Xiao-Lei Liu, Shun Yang, Zhao-Xia Luo, Liang Xie, Wen-Wen Zou, Jie-Ming Li and Shu Jiang 139true METHODS: BMSCs were isolated from 6-week-old Wistar Rat. GV303 virus were used as a IL-33 gene carrier to tansfect isolated BMSCs.Three days later, transfected BMSCs were observed under a fluorescence microscope. The expression level of IL-33 of transfected BMSCs was detected by Western blot and ELISA.

RESULTS: The BMSCs were successfully isolated, since flow cytometry results showed that rat BMSCs CD90 and CD29 positive, CD45,CD34 and CD11b negtive. Furthermore, BMSCs were able to be differentiated to osteoblasts, adipocytes and chondrocytes respectively. Green fluorescence of GFP BMSCs was observed under the fluorescence microscope 3d after virus transfecting. Expression of IL-33 was detected by Western blot and ELISA in the transfected rat BMSCs.

CONCLUSION: A rat BMSC cell line expressing IL-33 was established by GV303 virus transfection.]]> 2017/4/25 17:20:01 Xue-Yan Yao,Wei-Yang,Hai-Bo Li and Hui-Zhuo Xu 138true METHODS: Totally 60 healthy female SD rats were randomly divided into three groups: blank group, model group and experimental group, 20 rats in each group. Acute injury model of optic nerve in rats are established by crushing optic nerve. After the model of optic nerve injury was made successfully in model group, the rats were injected with balanced salt solution 5μL into vitreous cavity, and at 1 and 2wk later respectively they were injected again. In experimental group, after the model of optic nerve injury was made successfully, immediately the rats were injected with the PEDF 5μL into vitreous cavity, also at 1 and 2wk later respectively they were injected again. To observe the retinal morphologic changes and count the number of retinal ganglion cells, the specimen was given HE staining and observed under light microscope. The immunohistochemical semi quantitative method was used to detect the expression of Bcl-2 and Bax to observe the effect of PEDF on the optic nerve after injury.

RESULTS: Cell morphology of ganglion cells in experimental group was better and the number of RGCs was much more, comparing with the model group by HE staining. The expression of Bcl-2 and Bax in the experimental group was significantly different with that in the blank group and the model group(P<0.05). The expression of Bcl-2 increased and the expression of Bax decreases in the experimental group compared with the model group.

CONCLUSION: PEDF can inhibit or weak the apoptosis of RGCs cells after optic nerve injury and alleviate the injury of optic nerve by increasing the expression of Bcl-2 protein or decreasing the expression of Bax protein.]]> 2017/4/25 17:20:01 Jun-Bo Zhao,Wei-Xing Pu,Nan Huo and Yu-Quan Ma 137true METHODS: HLE-B3 cells were exposed to EF at 100mV/mm, 200mV/mm and 400mV/mm, respectively. Cells without EF-exposure were treated as normal controls. Photos of HLE-B3 cells were captured before and after EF-exposure, and the cell numbers were calculated. Apoptosis and cell cycle were detected with flow cytometry after EF-exposure for 24h.

RESULTS: After exposure to EF at 400mV/mm for 3h, HLE-B3 cells showed directed migration to the cathode. The cell number of HLE-B3 cells decreased gradually with continuous EF-exposure, and decreased by 12.6% at 6h and by 18.6% at 12h, respectively(P<0.05). After exposure to EF at 100mV/mm, 200mV/mm and 400mV/mm for 24h, the apoptosis rates of HLE-B3 cells increased dramatically compared to that of the normal control, which were(9.2±1.9)%,(23.9±2.6)% and(54±2.5)%, respectively(P<0.05). Accordingly, the results of flow cytometry showed that cell numbers in G2/M phase were increased after EF-exposure, which were(13.8±2.2)% and(15.6±2.5)% at 200mV/mm and 400mV/mm, respectively(P<0.05).

CONCLUSION: Applied direct EF induces directed migration of HLE-B3 cells and inhibits the cell growth with the prolongation of the EF-exposure time. EF also induces cell apoptosis and G2/M phase inhibition of the cell cycle in HLE-B3 cells.]]> 2017/3/27 10:17:04 Jing Han,Jun Liu,Hong Yan and Xiao-Long Yan 136true METHODS: Experimental rats were randomly divided into the control and experimental groups. Ten rats in the control group were bred in an indoor environment with the altitude of 1,500 meters; 60 in the experimental group were divided into 6 groups with each group of 10 rats, which were bred in a simulated experimental cabin of a simulated plateau environment with the altitude of 5,000 meters for 2, 6, 12, 24, 48 and 72h, respectively. After rats were killed in all groups, pathomorphological changes of rats' retinas were observed with hematoxylin eosin staining, expressions of caspase-3 and p53 in rats' retinas were noticed with immunohistochemical staining, and flow cytometry was used to detect early apoptosis rates.

RESULTS: The simulated hypoxic conditions at high altitude could cause rats' retinal tissues damage, and the longer hypoxic conditions were, the more obvious retinal pathological damage was. Expressions of caspase-3 and p53 were detected at 2h, and gradually increased with oxygen lack time increasing, the differences showed statistical significance(P<0.05). Early apoptosis rates of retinal cells gradually rose with oxygen lack time, and rose obviously at 48h.

CONCLUSION: Apoptosis may involve in the mechanism of rats' retinal damage induced by stimulated hypoxic conditions at high altitude, and play this role by the caspase-3-dependent apoptotic pathway.]]> 2017/3/27 10:17:04 Qian Zhang,Wen-Fang Zhang,Xiao-Hua Ji,Yi Yang and Yu-Ting Li 135true METHODS: The DM model was induced by intraperitoneal injection(IP)of streptozotocin in adult male rats, and the solution of RU486 was configured with dimethyl sulfoxide(DMSO). RU486 treatment group with glucocorticoid receptor antagonist RU486 and diabetic group with DMSO by intraperitoneal injection was in successful DM model. Naive rats were injected with DMSO as control group. Three months later, we detected the body weight and blood glucose and GC concentration of serum. The changes of retinal ganglion cell(RGC)density was investigated by HE staining. The expression of growth associated protein-43(GAP-43, a marker of neuronal axon regeneration)and synaptophysin(SYN, a marker of synaptic number)were semi-quantity analyzed by the optical density of immunofluorescence and Western blot.

RESULTS:Compared with the control group, the body weight and density of RGC and expression of SYN in diabetic group were significantly lower(P<0.01), the blood glucose and GC concentration of serum and expression of GAP-43 in diabetic group were significantly higher(P<0.01). Compared with the diabetic group, the density of RGC and expression of GAP-43, SYN in RU486 group were significantly higher(P<0.01).

CONCLUSION: Inhibition of GC could ameliorate the axonal degeneration of retinal neurons in diabetic rats, and loss of the number of synapses, and restore the RGC density of retina. The results suggest that long-term elevation of GC may be involved in the occurrence and development of diabetic retinopathy.]]> 2017/2/27 10:55:37 Wen-Qiang Liu,Yu-Bo Wang,Zhong-Fu Zuo,Hui-Min Liang,Zhao-Wei Li,Zheng Li and Xue-Zheng Liu 134true in vitro and in vivo and to assess the probability of defensins as a new application for infectious corneal diseases in the future.

METHODS: The synthetic rBD-2 DNA fragment was inserted between the XhoI and BamHI restriction enzyme cutting sites of eukaryotic expression vector pIRES2-ZsGreen1 to construct the recombinant plasmid pIRES2-ZsGreen1-rBD-2, then transformed it into E.coli DH5α, positive clones were screened by kanamycin and identified with restriction endonucleases and sequencing analysis. Transfection into the rat corneal epithelial cells was performed by lipofection. Then the experiment was divided into three groups: rat corneal epithelial cell was transfected with the recombinant plasmid pIRES2- ZsGreen1-rBD-2, rat corneal epithelial cell was transfected with the empty plasmid pIRES2-ZsGreen1 and the non-transfected group. The inverted fluorescence microscope was used to observe the transfection process. At last, the level of rBD-2 mRNA expressed in the transfected cells and the control groups are compared by the real-time fluoresence relative quantitative PCR.

RESULTS: The recombinant eukaryotic expression vector of pIRES2-ZsGreen1-rBD-2 was successfully constructed. The level of rBD-2 mRNA in transfected cells was significantly higher than that in control groups through the real-time fluorescence relative quantitative PCR.

CONCLUSION: The recombinant eukaryotic expression vector pIRES2-ZsGreen1-rBD-2 could be transfected into rat corneal epithelial cells, and exogenous rBD-2 gene could be transcripted into mRNA in the transfected cells.]]> 2017/2/27 10:55:37 Jing Dan,Xiu-Sheng Song,Yan-Ning Yang and Xia Li 133true METHODS: Rabbit lacrimal gland epithelial cells isolated and cultured for a period of time were separately cultured and mixed with isolated peripheral blood lymphocytes in a ratio of 1: 1 and cultured with 5-Bromo-2-deoxyUridine(BrdU)to detect the proliferation of peripheral blood lymphocytes, and the activated autologous peripheral blood lymphocytes were transferred to donor rabbits via ear vein. The rabbits were used as the rabbit model of autoimmune dry eye and the normal rabbits which did not induce the disease as the control group. Before and after the experiment, tear film rupture time and tear secretion at 2,4,6,8wk,were observed. At 4 wk after the transfusion the rabbits were sacrificed for the collection of bilateral upper and lower lacrimal gland and conjunctiva, and pathological HE staining, and then extract the total RNA in lacrimal gland. The expression of IL-17, TNF-α, IL-6 and TGF-γ was detected.

RESULTS: BrdU detected peripheral lymphocyte proliferation rate of 3.72. Compared with the normal group, the time of rupture of the tear film decreased(P<0.05), and the tear of the model group was significantly lower than that of the control group(P<0.05). The expression of IL-17, TNF-α, TGF-γ and IL-6 in the model group were significantly higher than those in the control group(P<0.05). The expression of IL-10 and TGF-β was significantly lower than those of the control group(P<0.05). The ratio of CD4⁺ CD2⁺ Treg cells in lacrimal gland tissue and spleen was higher in control group. The proportion of CD4⁺CD25⁺ Treg cells in model group decreased(P<0.05).

CONCLUSION: IL-17, TNF-α, IL-6 and TGF-γ play important roles in inflammatory reaction of lacrimal gland and the immunodepression active by CD4⁺CD25⁺Treg cells decreases when the rabbit model of dry eye is involved.]]> 2017/2/27 10:55:37 Feng Zhu and Feng Ke 132true METHODS: RGC-5 cell were divided into 3 groups: control group(DMEM high glucose medium+10% fetal calf serum), high glucose group(DMEM high glucose medium+10% fetal calf serum+30mmol/L glucose),NEP1-40 group(DMEM high glucose medium+10% fetal calf serum+30mmol/L glucose+1μmol/L NEP1-40). Detections were performed after 3d culture: the state of cell growth was observed by microscopy. Cell viability was detected by CCK-8 kit. The apoptosis rate of RGC cells was detected by flow cytometry(FCM). The intensity of ROS of the cells were detected by fluorescence microscopy. Intracellular MDA levels and SOD activity were measured by related kits. Western blot was used to detect the expressions of Bcl-2 and Bax proteins.

RESULTS: Compared with control group, high glucose group had a poor state and cell viability decreased, cell apoptosis rate significantly increased, ROS and MDA levels were significantly enhanced, SOD activity decreased, and the expression of anti-apoptotic protein Bcl-2 was decreased and the expression of pro apoptotic protein Bax was up-regulated. Compared with glucose group, after NgR expression was inhibited by NEP1-40, the oxidative stress reaction was reduced, Bcl-2/Bax was increased, the cell status was improved, the cell viability was increased, and the apoptosis rate was decreased in the NEP1-40 group(P<0.05).

CONCLUSION: High concentration of glucose can induce apoptosis of RGC-5 cells by NgR mediated oxidative stress reaction.]]> 2017/1/20 11:21:30 Yu-Bo Wang, Wen-Qiang Liu, Zhong-Fu Zuo and Xue-Zheng Liu 131true METHODS: Primary culture of RVEC of Wistar rats with 1-3d of birth,Ⅷ factor antibody immunohistochemistry to identify the cells; Establishment 0mmol/L glucose as a control group(group C), at different concentrations of glucose intervention in turn divided into 10mmol/L(group A), 15mmol/L(group B), 25mmol/L(group D), 35mmol/L(group F), MTT assay was used to detect cell OD values of each group and each group was detected by flow cytometry apoptosis rate were at three time points: 24,48,72h。

RESULTS: RVEC purity was 94%. Effect of different concentrations of glucose in Wistar rat RVEC by MTT assay: after 24h culture, OD value of experimental group decreased, but the difference was not statistically significant(F=0.42, P=0.7411); after 48h culture, OD value of the difference between the groups was statistically significant(F=45.2, P=0.000), pairwise comparison between the five groups, except among groups A, B, C group no difference was not statistically significant external(Q_48AB=44.0427, Q_48AC=36.4701, Q_48BC=27.7953, all P> 0.05), the remaining pairwise comparisons were statistically significant difference between groups(Q_48CD=11.0715, Q_48AD=14.0794, Q_48AF=12.2964, Q_48BD=23.4698, Q_48BF=12.7016, Q_48DF=10.2013, all P<0.05; Q_48CF=6.4701, P<0.01). After 72h culture, OD value of each group compared to the difference was statistically significant(F=37.46, P=0.000), pairwise comparisons among five groups, the same A, B, C no difference among the three groups was not statistically significant(Q_72AB=27.7338, Q_72AC=25.0054, Q_72BC=33.3797, all P>0.05), the rest of the group differences were statistically significant(Q_72AD=13.4793, Q_72BD=12.7546, both P<0.05; Q_72CD=7.3743, Q_72CF=8.3465, Q_72AF=4.7455, Q_72BF=3.9471, Q_72DF=3.2649, all P<0.01). Effect of different concentrations of glucose on RVEC apoptosis rate: after 24h culture, Differences between groups was not statistically significant(P>0.05). After 48h culture, pairwise comparison among five groups, Q_48AB=31.1704, Q_48AC=33.5947, Q_48BC=29.3968, Q_48AD=30.4097, Q_48BD=28.8164, both P>0.05; Q_48CD=12.5032, Q_48AF=11.7531, Q_48BF=14.1076, Q_48DF=13.9372, both P<0.05; Q_48CF=7.0953, P<0.01. After 72h culture, pairwise comparison among five groups, Q_72AB=26.3392, Q_72AC=24.9142, Q_72BC=30.2976, both P>0.05, the rest of the group differences were statistically significant(where Q_72AD=14.1983, Q_72BD=12.9356, both P<0.05; Q_72CD=6.8752, Q_72CF=7.6745, Q_72AF=5.0545, Q_72BF=4.0741, Q_72DF=3.8876, both P<0.01).

CONCLUSION: Best glucose concentration and the time is 25 mmol/L and 48h, for RVEC high sugar model.]]> 2016/12/21 10:42:28 Feng-Jiu Zhang, Li-Min Zhang, An-Ling Lin, Xiang-Dong Peng, Bing Liu, Jian-Ling Yang, Ling-Yan Yu and Hai-Ming Wang 130true in vitro.

METHODS:Human RPE cells were cultured and passaged, and 3-6 generation cells were used in the experiment. The experiment was divided into normoxia control group and hypoxia group. The human RPE cells in normoxia control group were routinely cultured. The culture medium containing 200μmol/L CoCl₂ was used to establish the hypoxia model of human RPE cells cultured in vitro for 0,4,6,8,12 and 24h, and the RPE cells cultured normoxia were as controls. The effects of different time of hypoxia on the expression of NOX4 were identified by Immunofluorescence staining and Western-blot assay.

RESULTS:RPE cells grew well in the control group and hypoxia group. The morphology was spindle. The morphology and arrangement of cells in hypoxia group had no significant change compared with the control group. Immunofluorescence staining showed that there was a small amount of NOX4 expression in the RPE cells of the normoxia control group. Cytoplasm showed green fluorescence while the nucleus was blue. The expression of NOX4 increased after hypoxia, and continued until the late stage of hypoxia. Western-bolt results showed that the expression level of NOX4 under hypoxic conditions in RPE cells increased significantly, and is proportional to the hypoxia time, the difference was statistically significant(P<0.05).

CONCLUSION: NADPH oxidase 4 in normal human RPE cells have a small amount of expression, hypoxia can significantly increase the expression of NOX4, and is proportional to the time. It suggests that NADPH oxidase may play an important role in the formation of choroidal neovascularization.]]> 2016/12/21 10:42:28 Jing Li, Jing Yang and Qian-Yan Kang 129true METHODS: Totally 40 three-month old healthy SD rats which had no eye diseases, were randomly assigned into two groups: simvastatin group(n=20)and control group(n=20). All rats were cultivated under the same conditions until they were nine-month old when interventions started to be given. Simvastatin group was given intragastric administration of 5mg/kg simvastatin every day until 17-month old. Control group was given intragastric administration of same amount of saline gavage. Retinal thickness was measured by HE staining, while Cu-Zn-SOD, NOX2, Bcl-2 and Bax were determined by immunohistochemistry(IHC).

RESULTS:HE staining showed that the retinal structure was clearer; the morphology of cell was more homogeneous; the number of cells was more stable and the structure of retinal pigment epithelium was more compact when compared with control group. Thickness of retinal neuroepithelium layer and retinal pigment epithelium increased significantly in the simvastatin group. Immunohistochemistry analysis showed that the expressions of Cu-Zn-SOD and Bax statistically increased while the NOX2, Bcl-2 as well as Bcl-2/Bax decreased in simvastatin group when compared with control group(P<0.05).

CONCLUSION: Simvastatin plays a protective role in retinal aging by decreasing oxidative stress reaction and promoting cell apoptosis.]]> 2017/11/20 16:09:09 Jiang Zhu, Ye-Wen Shi and Qian-Yan Kang 128true METHODS: The rat model of corneal neovascularization(CNV)was induced by corneal suture. Rats were randomly divided into Group A: physiological saline control group containing DMSO(10 rats); Group B: 25μmol/L DATS treatment group(10 rats); Group C: 50μmol/L DATS treatment group(10 rats); Group D: 100μmol/L DATS treatment group(10 rats); Group E: 200μmol/L DATS treatment group(10 rats). The occurrence and development of CNV were observed by slit-lamp microscope at 7d after suture, and the area of CNV were calculated.Two weeks later, HE staining was used to observe the pathological organization form of each cornea, and RT-PCR and Western blot were used to detect the expression of VEGF mRNA and protein expression of VEGF and p-AKT between each groups.

RESULTS: The blood vessel area of Group C, D and E was compared with that of Group A, the difference was statistically significant(P<0.05); HE slice showed corneal edema, angiogenesis and inflammation infiltration situation gradually reduced comparing with the Group A, with the increase of concentration of DATS. RT-PCR showed the expression of VEGF mRNA in Group B, C, D, and E decreased compared with the Group A, and the difference was statistically significant(P<0.05). Western-blot showed that the expressions of VEGF and p-AKT in Group B, C, D and E decreased gradually compared with those in Group A, and the difference was statistically significant(P<0.05).

CONCLUSION: DATS can inhibit corneal neovascularization of the rats induced by suture. Its mechanism may be associated with suppression of VEGF secretion, down-regulation of VEGF and inactivation of p-AKT.]]> 2017/10/19 16:49:00 Xiao-Jun Zhou 127true in vivo and in vitro.

METHODS: We observed the biological characteristics of human corneal epithelial cells. The cell proliferation was analyzed using CCK-8 assay, which also used to test the toxicity of MIL60 and the solvent on cultured human corneal epithelial(HCE). FACs was used to analyze the apoptosis of HCE after treated with MIL60. Also we evaluated the effect of subconjunctival injection of MIL60 on corneal epithelial healing model in normal rat and rats with epithelium defect through slit lamp-microscopy, Draize scores and histopathology way.

RESULTS: The proliferation speed of HCE in three groups was the same. MIL60 did no harm on the proliferation of HCE and the apoptosis of HCE, and has no effect on corneal epithelial healing and other parts of the ocular in rats without inflammation cells infiltration.

CONCLUSION: When given subconjunctival injection, Mil60 does no harm to the proliferation and apoptosis of HCE, and is safe with ocular application.]]> 2017/10/19 16:49:00 Qun Wang, Hua Bai, Jie Zhao, Bao-Jie Hou, Yi-Fei Huang and Ming Lyu 126true METHODS: Rats were divided randomly into five groups: control group, glaucoma group, RSCs group, GKB group and RSCs combination therapy group. A chronic glaucoma model was established in rats, accordingly. The morphological changes in ocular tissues were analyzed by HE staining. Retinal ganglion cells apoptosis were analyzed by TUNEL staining. The protein expressions of Bcl-2, Bax, Cleaved caspase-3 and Cleaved caspase-9 were determined by Western blot. The mRNA levels of Bcl-2 and Bax were determined by qPCR.

RESULTS: HE staining revealed that RSCs transplantation or GKB treatment decreased fiber interstitial edema and vacuole, as compared to glaucoma group. Furthermore, this improvement was more pronounced in combination therapy group than in single treatment alone. Combination therapy significantly inhibited retinal ganglion cells apoptosis, increased Bcl-2 mRNA and protein expression,but decreased Bax mRNA and protein expression. Moreover, the protein expression of Cleaved caspase-3 and Cleaved caspase-9 expression were decreased after combination therapy.

CONCLUSION: Our data demonstrate that combination of Ginkgobalide B and retinal stem cells transplantation can inhibit retinal ganglion cells apoptosis and protect against glaucoma. These effects may be associated with the regulation of Bcl-2, Bax, Cleaved caspase-3 and Cleaved caspase-9 expression.]]> 2017/10/19 16:49:01 Xian Liu, Bing Wang and Ke Fu 125true in vitro and in vivo experiments.

METHODS: The rabbit's corneal epithelium were cultured in the lower chamber of transwell, and AEC suspension liquid was dropwised in the upper chamber. There was only culture medium in the upper chamber of the control group. The proliferation of rabbit's corneal epithelium was observed with CCK-8 automated colorimetry and the expression of PCNA was detected by immunocytochemistry. We used the scratch wound assay to detect the migration of corneal epithelial cell(CEC). The in vivo models were established by placing a 10mm diameter corneal trephine in the center of the cornea, within 1mol/L NaOH for 1min. We divided those into three groups: treatment group of AEC suspension liquid eye drop, AEC suspension liquid subconjunctival injection and the control group without any treatment. Using the slit-lamp biomicroscope and fluorescence staining to observe the cornea per week. After 28d we took the eyeballs with the HE staining. The expression of VEGF was detected by immunohistochemistry.

RESULTS: The activity of CEC with AEC treatment was much higher than the control group(P<0.05). The expression of PCNA increased in AEC group(P<0.05). And the migration of CEC in the AEC group was faster than the control one. In vivo, the inflammation of the corneal and the CNV of the AEC group were all significantly reduced compared with the control group(P<0.05). There were less invasive cells and more ordered organization arrangement in ACE group observed by the HE staining. The expression of VEGF and mcp-1 in these two AEC treatment groups all significantly decreased compared with the control group(P<0.05).

CONCLUSION: AEC suspension liquid can promote the proliferation and migration of the rabbit's corneal epithelium. The potential of AEC suspension liquid as a therapy for acute corneal alkali burn.]]> 2017/9/18 10:53:55 Yan-Yan Zhang, Hong-Ling Liu, Hong Zhang and Shao-Ying Fu 124true METHODS: RPE cells were cultured and divided into four groups according to randomized controlled method: blank control group: the cells were treated with 5.5mmol/L routine glucose medium for processing; high glucose group: cells were treated with 100mmol/L glucose for 12h; 17β-estradiol low concentration group: after treated with 10 μmol/L 17β-estradiol, cells were treated with 100mmol/L glucose for 12h; 17β-estradiol high concentration group: after treated with 100 μmol/L 17β-estradiol, cells were treated with 100mmol/L glucose for 12h. Cell viability were tested by MTT colorimetric detection. Cells apoptosis were detected by Hochest33258 staining. Intracellular reactive oxygen species(ROS)level were detected by H₂DCFDA staining. Expression of CAT, SOD and MDA were tested by colorimetric detection.

RESULTS: RPE cell activity decreased with the concentration of glucose increased; 17β-estradiol inhibited high glucose-induced cell viability decrease in RPE cells, decreased the apoptosis rate of RPE cells and intracellular ROS generation; besides, 17β-estradiol significantly increased the expression of CAT, SOD and decreased the expression of MDA in RPE cells.

CONCLUSION: The 17β-estradiol effectively inhibited high glucose -induced RPE cells damage, which provide reliable experimental basis for the treatment of injuries in RPE cells.]]> 2017/9/18 10:53:55 Meng-Yao Jiao, Yang-Yang Zhang, Xuan Sun, Kai Sun, Xu-Cong Kang, Wei Jiang and Na Chen 123true METHODS: Primary cells of RPE cells were obtained by enzymatic digestion, and the primitive culture and subculture of RPE cells were proceeded; prepared blood plasma-contained traditional Chinese medicine compound drug-containing plasma; the fourth genertation rabbit RPE cells were selected as the experimental cells by PDGF of low, medium and high doses(5μg/L, 10μg/L, 20μg/L)for 48h. Suitable concentration was detected and selected in cells experiment by using CCK-8 method. Establishing rabbit model of RPE cell proliferation treated with PDGF. The experimental groups were blank control group(DMEM), normal plasma group, PDGF(10 μg/L)group, PDGF(10 μg/L)+ AG1296(10 μmol/L)group, PDGF(10 μg/L)+ 10% drug-containing plasma, PDGF(10 μg/L)+ 20% drug-containing plasma. Respectively, transwell method was used to determine the migration of rabbit RPE cells after 24h intervention in each group; CCK-8 was used to determine the cell viability OD value of rabbit RPE cells after 48h of intervention in each group.

RESULTS: The plasma containing 10% and 20% concentration of invigorating blood and dissipating masses traditional Chinese medicine compound drug can effectively inhibit cell viability and cell migration of RPE cells treated with PDGF.

CONCLUSION: We found the certain concentration of invigorating blood and dissipating masses traditional Chinese medicine compound drug can effectively inhibit cell viability and cell migration of RPE cells treated with PDGF, which may be an effective treatment for proliferative vitreoretinopathy and provide a new way to study the mechanism of proliferative vitreoretinopathy.]]> 2018/8/17 17:07:06 Quan-Long Wu, Xiao-Qing Liu, Jun Peng, You-Wei Zhang, Kun Pan, Jian-Chao Li and Qing-Hua Peng 122true METHODS: The hUCMSCs were isolated and cultured, hUCMSCs were transfected with PEDF recombinant lentivirus. Experimental royal college of surgeon(RCS)rats were randomly divided into 3 groups, 8 rats in each group. The experimental group was injected with PEDF-hUCMSCs subcutaneously. The hUCMSCs control group was injected with the same amount of hUCMSCs. The PBS control group was injected with the same amount of PBS. At 4 and 8wk after injection, electroretinography(ERG), the thickness of retinal outer nuclear layer(ONL)and green fluorescent protein(GFP)staining were observed. Immunofluorescence staining of retinal sections and the phagocytosis of MERTK protein were evaluated to determine phagocytosis.

RESULTS: PEDF gene carrying recombinant lentiviral vector could efficiently infect hUCMSCs. After infection, hUCMSCs were sub cultured and the lentivirus prolonged the expression in the cells of the target gene. At 8wk after transplantation, the amplitude of b-wave in ERG were significantly higher in the experimental group than in the control groups. At 4 and 8wk after transplantation, morphological changes of ONL thickness in the experimental group were significantly higher than those in the control groups(all P<0.05). At 4 and 8wk after transplantation, hUCMSCs infected by lentiviral vector carrying PEDF were found survival in the subretinal cavity by GFP staining, and the PEDF-hUCMSCs were found having phagocytosis for outer segment fragments of photorecepter cells by immunofluorescence staining. At 4 and 8wk after transplantation, the Protein band gray values in experimental group were significantly higher than that in control groups, which indicated that the expressions of MERTK protein in experimental group were also increased significantly, and the phagocytic capacity of RPE were improved.

CONCLUSION: Transplantation of pigment epithelium-derived protein modified human umbilical cord-derived mesenchymal stem cells has a protective effect on the retina of RCS rats.]]> 2018/7/20 11:38:05 Hui Wen, Xiao-He Yan, Yuan-Fei Zhu, Long-Hao Kuang and Luo-Dan A 121true METHODS: RPE cells were cultured and divided into the negative control group, high sugar group, Ghrelin low dose group(10^-9 mol/L)and high dose group(10^-6 mol/L). Cells survival rate were detected by CCK-8 colorimetry, cells oxidative damage were observed by oxygen sensitive fluorescence probe H₂DCFDA staning, changes of intracellular reactive oxygen species(ROS)were detected by H₂DCFDA staining, super oxide dismutase(SOD)activity and malondialdehyde(MDA)content were detected by spectrophotometer colorimetry.

RESULTS: CCK-8 results showed that RPE cells survival rate increased to 54.79%±3.43% and 79.16%±3.29% after treated with 10^-9 mol/L, 10^-6 mol/L Ghrelin, the difference was statistically significant compared with high glucose group(41.65%±3.42%)(P<0.05). H₂DCFDA fluorescent probe dying showed that Ghrelin reduced ROS generation in RPE cells and decreased oxidative damage cells. Spectrophotometer colorimetric method showed that according to the high sugar group, SOD activity increased and MDA content decreased in Ghrelin group.

CONCLUSION: Ghrelin could inhibit high glucose-induced oxidative damage in human RPE cells, which has protective effect on the process of the occurrence and development of diabetic retinopathy.]]> 2018/6/27 10:52:29 Li-Li Hou, Ting Chen, Li-Ting Liu and Li-Juan Zhang 120true METHODS: We performed anterior chamber injection of magnetic beads to induced chronic ocular hypertension models. Adult C57BL/6 male mice were used in the experiments and randomly divided into four groups: control, Beads group, E2 group and E2+TAM group. The intraocular pressure(IOP)were measured by Tonolab tonometer. Central retinal thickness was evaluated by HE staining. Brn3a as a specific marker of retinal ganglion cells(RGCs), were stained and counted by immunohistochemistry(IHC)staining. Glial fibrillary acidic protein(GFAP)as a marker of proliferation of astrocytes, was quantified using western blotting.

RESULTS: The IOP level was significantly elevated after anterior chamber injection of magnetic beads compared to control group(P<0.05), while in E2 and E2+TAM group, the IOP levels were reduced(P<0.05 vs Beads group), especially in E2+TAM group in 2wk. The RGCs happened to degenerated in Beads group after 2wk, while the effects were reversed by E2+TAM(P<0.05). The central retinal thickness showed no significant statistical difference among the four groups after 2wk(P>0.05). The expression level of GFAP increased caused via beads injection, however, decreased in E2 and E2+TAM group(P<0.05).

CONCLUSION: E2 and E2+TAM could both effectively decrease the IOP level in chronic intraocular hypertension mouse model, increase the survival of RGCs from high intraocular pressure, suppress expression of GFAP, which indicated neuroprotective effects of E2 and TAM in glaucoma through an anti-inflammatory effects.]]> 2018/5/25 15:46:38 Xue-Yun Ma, Qin-Qin Deng, Yin Shen and Yi-Qiao Xing 119true METHODS: A total of 50 male Sprague-Dawley(SD)rats were randomly divided into normal control group, and 3d, 1, 4, 8wk group after acute elevated intraocular pressure(IOP)(n=10 per group). Acute intraocular hypertension model was established by anterior chamber perfusion of normal saline in the right eye. The expression levels of microtubule-associated protein 1 light chain 3(LC3)was measured by immumofluorescence method. To determine whether autophagy and paraptosis were activated. Retinal sections were examined by transmission electron microscopy(TEM). Autophagosomes and cytoplasmic vacuoles in the cytoplasm of RGCs were measured.

RESULTS: TEM analysis revealed that double- and multiple-membrane vacuoles containing electron-dense materials of autophagosomes were found in RGCs. The number of autophagosomes per 50μm² were 0.79±0.43, 2.14±0.36, 2.29±0.47, 1.57±0.51 and 1.21±0.43 in the normal control group and in acute IOP group at 3d, 1wk, 4wk, 8wk, respectively. The number of autophagosomes markedly increased in the cytoplasm of RGCs at 3d, 1wk, 4wk, 8wk groups than those in the normal control group(all at P<0.05). LC3 positive expression was rarely detected in ganglion cell layer(GCL)in the normal control group and percentage of LC3 positive cells was 15.90%. Immumofluorescence analysis showed that the percentage of LC3 positive cells statistically increased in acute IOP groups when compared with control group(P<0.05). The number of RGCs per 200μm in each group of acute IOP injury significantly decreased compared with the normal control group(P<0.05). Cytoplasmatic vacuolization were observed in RGCs at 3d after acute IOP injury and lasting to 8wk. TEM also revealed that a large number of cytoplasmic vacuoles were derived predominantly from the progressive swelling of mitochondria and/or endoplasmic reticulum(ER).

CONCLUSION: Autophagy and paraptosis participate in the death of RGCs under transiently elevated intraocular pressure. Different types of programmed cell death(PCD), coexistence of multiple cell death forms or a single cell death form, participates in the pathogenesis of acute elevation of intraocular pressure.]]> 2018/5/25 15:46:38 Ting Wei, Shan Gao, Bo Ma, Ning Gao and Qian-Yan Kang 118true METHODS: The mRNA was isolated from venous blood of 14 congenital cataract patients with autosomal dominant inheritance in 3 chromosomes. Based on genetic mutats of CRYAB gene, DNA probe, capture probe and signal probe were designed. The sandwich structures containing capture probe, DNA probe and signal probe was used to detect genetic mutants in 8 samples from one family; and ELISA was used to detect the contents fluctuation of Crystallin alpha B.

RESULTS: The dual-specific probe technique detected the minimum genetic mutation of Crystallin alpha B in congenital cataract samples, various mutations detection rate was between 99.5% and 99.7%. In ELISA detection, serum Crystallin alpha B level increased, and detection rate was 85.9%. Compared with ELISA assay, this novel assay was more sensitive.

CONCLUSION: The novel dual-specific probe method is quite convenient for detection of genetic mutants of congenital cataract; and for its high sensitivity and repeatability, it is of great potential in clinically prenatal diagnosis, and it might play significant roles in eugenic and superior nurture.]]> 2018/5/25 15:46:38 Yu-Juan Cao, Jun Shao, Ying-Qing Yu, Yong Yao, Jing Zhu, Yu-Jing Tian and Jia Cao 117true Rhodiola sachalinensis on ocular blood flow in diabetic retinopathy rats.

METHODS: A total of 90 SD rats were randomly divided into control group(n=30), model group(n=30)and intervention group(n=30). Rats in the model group and intervention group were fed with high glucose and high fat diet and injected with streptozotocin(40mg/kg)in order to construct DR model rats, while rats in the control group were fed with basic diet and injected with the same amount of normal saline. After 4wk, the rats in the intervention group were injected with Rhodiola sachalinensis injection(10mL, one per day), while rats in the control group and the model group were injected with normal saline. The course of intervention treatment was 4wk. The ocular blood flow in rats was detected by laser doppler flowmetry, the apoptosis of retinal cells in rats was detected by TUNEL method, the expression of vascular endothelial growth factor(VEGF), glial fibrillary acidic protein(GFAP)and glutamate/aspartate transporter(GLAST)in retina of rats was detected by RT-PCR method and Western blot method.

RESULTS: The whole blood viscosity, whole blood high shear viscosity, low shear blood viscosity and erythrocyte sedimentation of rats in the model group and intervention group were higher than those in the control group(P<0.01), while the indexes of intervention group were lower than that of model group(P<0.01). The PSV and EDC of rats in the model group and intervention group were lower than those in the control group(P<0.01), and the RI was higher than that in control group(P<0.01). The PSV and EDC in the intervention group were higher than those in the model group(P<0.01), and RI was lower than that in the model group(P<0.01). The apoptosis rate of retinal cells of model group and intervention group was higher than that of the control group(P<0.01), that in the intervention group was lower than that in the model group(P<0.01). The expression of VEGF and GFAP in retina tissue of model group and intervention group was higher than that of the control group(P<0.01), and the expression of GLAST was lower than that in the control group(P<0.01). The expression of VEGF and GFAP in the intervention group was lower than that in the model group(P<0.01), and the expression of GLAST was higher than that in the model group(P<0.01).

CONCLUSION: Rhodiola sachalinensis injection can significantly improve the ocular blood flow in diabetic retinopathy rats, reduce the apoptosis of retinal cells in rats, down regulate the expression of VEGF and GFAP, and up regulate the expression of GLAST.]]> 2018/4/24 14:27:15 Ai-Ping Gu, Yi Wu and Bing-Hua Yang 116true METHODS: The Y79 cells were divided into three groups: blank control group, negative control group and TRAF6 siRNA group. After TRAF6 siRNA transfection, the levels of TRAF6 mRNA and protein in Y79 cells were examined by RT-PCR and Western blotting. MTT assay was used to detect cell proliferation. Flow cytometry was employed to detect changes in cell cycle and apoptosis. Cell invasiveness was detected by the Transwell method.

RESULTS: Expression of TRAF6 mRNA and protein in the TRAF6 siRNA group significantly decreased compared with the negative and blank control groups. Following the silencing of TRAF6, cell proliferation was inhibited and the apoptosis rate increased; the cell cycle was arrested at G0/G1 phase; the number of cells in S phase was reduced, while the invasion ability of cancer cells decreased.

CONCLUSION: Silencing TRAF6 may inhibit the proliferation of Y79 cells, promote cell apoptosis, arrest the cell cycle at G0/G1 phase and decrease the invasive ability. Thus, TRAF6 may be a potential target in therapy for retinoblastoma.]]> 2018/4/24 14:27:15 Yu-Shun Xue, Rui Shi, Le Yang, Hai-Yan Zhou, Li-Ping Chen and Rong Chai 115true METHODS: Real-time quantitative PCR(RT-qPCR)was used to detect miR-138 expression in the anterior lens capsules of healthy people, the anterior lens capsules of patients with age-related cataracts, and human epithelial cell line(SRA01/04)cells exposed to oxidative stress. A 2', 7'-dichloro-fluorescein diacetate(DCFH-DA)probe was used to measure the levels of endogenous reactive oxygen species(ROS)in human lens epithelial cells(hLECs)exposed to 400μmol/L H₂O₂ for 1h. SRA01/04 cells were transfected with either miR-138 mimics, mimic controls, miR-138 inhibitors or inhibitor controls. After 72h, these cells were exposed to 400μmol/L H₂O₂ for 1h, then p53 and Bax mRNA expression were measured using RT-qPCR. Expression of p53 and Bax protein were also measured by western blotting analysis. Finally, cell viability was assessed using an MTS assay.

RESULTS: Compared to the control group, expression of miR-138 in the anterior lens capsules of age-related cataract patients and in SRA01/04 cells exposed to oxidative stress significantly increased(P<0.001). Levels of endogenous ROS were significantly elevated in hLECs exposed to oxidative stress(P<0.001). Compared to the mimic control group, the hLECs in the miR-138 mimic group expressed significantly higher levels of p53 and Bax mRNA and protein while cell viability was significantly reduced(P<0.001). Conversely, p53 and Bax mRNA and protein expression were significantly reduced in the miR-138 inhibitor group as compared to the control group, while the cells in this group had much higher levels of cell viability(P<0.001).

CONCLUSION: The expression of miR-138 is upregulated in the anterior lens capsules of age-related cataract patients. MiR-138 decreases the anti-oxidative stress capacity of lens epithelial cells by upregulating p53 and Bax, while inhibiting cell proliferation and repair. This finding suggests that miR-138 may play a key role in the development of age-related cataracts.]]> 2018/3/26 15:30:52 Bo Lu, Li-Wei Ma, Xin-Ling Wang, Xiao Han, Li Feng, Ling-Feng Jiang, Chun-Xia Wang, Jin-Song Zhang and Qi-Chang Yan 114true METHODS: Twelve 7-day-old(P7)C57BL/6J mice were randomly divided into normal group, OIR group and OIR+VX-765 group. OIR models were established in OIR group and OIR+VX-765 group. Caspase-1 inhibitor VX-765(4mg/kg, dissolved in 0.4% polyethylene glycol)or 0.4% polyethylene glycol, were intraperitoneally injected from P12 to P16 into the mice of OIR+VX-765 and OIR groups, respectively. Whole retinal flatmounts of P17 mice were prepared, and Lectin staining was performed to calculate the ratio of avascular and neovascular area to retina area. The frozen sections of the posterior ocular segment were prepared, and the distribution of Caspase-1 and activated microglial cells were detected by immunofluorescence technique. Cultured BV-2 cells were divided into control group, hypoxia group and inhibitor group. The cells of inhibitor and hypoxia groups were pre-treated with VX-765 or 0.4% polyethylene glycol for 3h, and then hypoxic incubated for 24h. The expression levels of Caspase-1, p20(active form of Caspase-1), IL-1β and VEGF were detected by Western blotting. The angiogenesis and migration capacity of cultured RF/6A cells were assessed by endothelial cell tube formation assay and migration assay, after they were incubated with supernatant of those different BV-2 groups.

RESULTS:The distribution and morphology of retinal blood vessels were normal in P17 mice of the normal group, and avascular and new blood vessel cluster were found in the mice of OIR group and OIR+VX-765 group. The ratio of avascular area was 12.23%±1.02% and that of the new blood vessel area was 2.16%±0.52% in the OIR+VX-765 group, which decreased in comparison with 16.58%±1.14% and 4.00%±0.41% of the OIR group(P<0.01). Caspase-1 was rarely detected by immunofluorescence staining in the normal retina of the mice, whereas it was mainly co-located with activated microglial cells in the ganglion cell layer and the inner plexiform layer in the mice of OIR group. The expression of Caspase-1, p20, IL-1β and VEGF increased in BV-2 cells of the hypoxia group, which were down-regulated by VX-765(P<0.05), except Caspase-1. The tube length was 271±12, and the number of migrated cells was 347±34 in RF/6A cells cultured with supernatant of BV-2 cells in the hypoxia group, which significantly decreased to 171±22 and 212±27 with inhibitor of Caspase-1(P<0.05).

CONCLUSION:Caspase-1 promotes retinal neovascularization in the mice with OIR, probably by activating the downstream inflammatory factor IL-1β in microglial cells and accelerating the release of VEGF.]]> 2018/3/26 15:30:52 Zhi-Cha Hu, Yu-Sheng Wang, Wen-Qin Xu and Zi-Feng Zhang 113true 2 (TGF-β₂).

METHODS: The HTFs were identified by immunofluorescence analysis with Vimentin and keratin. HTFs with no other addiction was as normal control; H+T group: HTFs+5μg/L TGF-β₂; H+T+NC SiRNA group: HTFs+5μg/L TGF-β₂+50nmol/L negtive SiRNA; H+T+ILK SiRNA group: HTFs+5μg/L TGF-β₂+50nmol/L ILK SiRNA. The ILK SiRNA were transfected into HTFs by lipofectamine 2000, then the cells were stimulated with 5μg/L TGF-β₂. The protein expression of ILK were analyzed by Western Blot. The proliferation levels of HTFs were analyzed by CCK-8, the apoptosis of HTFs were analyzed by Hoechst 33342/PI double staining.

RESULTS: The protein ILK were expressed in both TGF-β₂ treated and control groups, and TGF-β₂ up-regulated the expression of ILK, ILK SiRNA inhibited the protein expression of ILK(P<0.05). CCK-8 analysis showed that compared with the normal control group, the cell proliferation rate of HTFs in TGF-β₂ treated group increased, and in ILK SiRNA group the cell proliferation rate was suppressed after exposured to ILK SiRNA for 48h(P<0.05). Hoechst 33342/PI double staining showed that there was no change on the apoptosis of TGF-β₂ stimulated group(P>0.05), compared with the normal control group, however in the ILK SiRNA group, we found lots of apoptosis cells and a few of necrotic cells(P<0.05).

CONCLUSION: The ILK SiRNA attenuates the abnormal proliferation of HTFs induced by TGF-β2, thereby enhancing the apoptosis of HTFs.]]> 2018/3/26 15:30:53 Qun Wang, Li-Jun Cui, Si-Wei Liu and Qian-Yan Kang 112true in vitro.

METHODS: The concentration of recombinant myocilin protein was 0(control group), 0.5, 1, 1.5, 2 and 2.5μg/mL, respectively. The expression of FN in the cells and the cell culture medium were detected by Western blot and ELISA. The effect of myocilin protein on the adhesion of cultured POAG trabecular meshwork cells was detected by CCK-8 method.

RESULTS: Western blot showed that the ratio of FN/β-actin was 34.8±0.6, 33.4±1.0, 28.9±0.8, 21.6±0.9, 15.9±1.1 and 11.9±0.8; that of 0.5μg/mL group was not significantly different compared with that of control group(P=0.092); that of 1.0, 1.5, 2 and 2.5μg/mL group were significantly different compared with that of the control group(F=346.131, P<0.05). The concentration of FN in the cell culture medium was 0.4654±0.0039, 0.4596±0.0032, 0.4216±0.0037, 0.4214±0.0039, 0.4043±0.0039, 0.3806±0.0071μg/mL by ELISA; difference between that of 0.5μg/mL group and that of control group was not statistically significant(P=0.120); the difference between of 1, 1.5, 2, 2.5μg/mL group and the control group were statistically significant(F=176.054, P<0.05). The mean values of the absorbance values of each group were 1.9814±0.1624, 1.8848±0.0267, 1.4895±0.0916, 1.4120±0.1087, 1.3392±0.1391, 1.0310±0.0639 through CCK-8 method; that of 0.5μg/mL group was not significantly different compared with that of control group(P=0.300); the difference between of 1, 1.5, 2, 2.5μg/mL group and the control group were statistically significant(F=177.818, P<0.05).

CONCLUSION: Myocilin protein can inhibit the fibronectin expression in POAG trabecular meshwork cells, showing a dose dependent manner. Myocilin protein can reduce the adhesion of POAG trabecular meshwork cells.]]> 2018/2/27 0:00:00 Yi-Hong Huang and Yu-Yu Wu 111true [THIS ARTICLE HAS BEEN RETRACTED]AIM: Through the expression of VEGF₁₆₅ and VEGF_165b in human retinal pigment epithelial cells in vitro in artificial simulated hypoxia and high glucose environment, to discuss their roles in the development of diabetic retinopathy and the relationship between each other.

METHODS: After normal inoculation and cultivation of human retinal pigment epithelial cells(RPE)in vitro, the cells was divided into the normal group(5.56mmol/L glucose, without CoCl₂), the hypoxia group(5.56mmol/L glucose + 150μmol/L CoCl₂), the high glucose group(25mmol/L glucose, without CoCl₂), the combination group(25mmol/L glucose + 150μmol/L CoCl₂), a total of four groups. The RNA of each group was extracted respectively in 12h, 24h, 36h, and 48h. We used the MTT colorimetry to detect cell vitality and growth trend; RT-PCR method to detect VEGF₁₆₅ and VEGF_165b relative expression of mRNA of RPE cells in four different time points.

RESULTS: Hypoxia and high sugar environment limited proliferation of RPE cell division and cell vitality. After comparing cells of the same group in different time points, in the normal group there was no statistically significant different expression over time(P>0.05); the expression in the hypoxia group, the high glucose group and the combination group increased over time, the difference was statistically significant(P<0.05). At the same time, differences of the expression between groups was not statistical significant in 12h(P>0.05); the difference was statistically significant in 24h, 36h, 48h(P<0.05).

CONCLUSION: Cultured RPE cells can express VEGF_165b normal. Lack of oxygen and high glucose can induce the increase of VEGF₁₆₅ mRNA, at the same time reduces the VEGF_165b mRNA expression.]]> 2018/2/27 0:00:00 Ya-Jing Niu and Shui-Qing Hu 110true METHODS: Sixteen healthy New Zealand white rabbits with 6wk old received corneal lamellar keratoplasty, and the corneal graft was ostrich acellular corneal stroma. After surgery all subjects were divided into two groups, Group A(experimental group)were administrated with subconjunctival injection of compound betamethasone injection(once every 7d), and Group B(control group)were administrated with subconjunctival injection of dexamethasone sodium phosphate(once every 7d). At 1,2wk, 1, 2mo after the surgery, rabbit corneas were taken for paraffin sections, and were observed with H-E staining, in the meantime changes of CD4⁺ and CD8⁺ T lymphocytes were observed by immunofluorescence.

RESULTS: Two months after surgery, in Group A corneal grafts remained transparenct, and showed little neovascularization; HE staining and indirect immunofluorescence showed that only a few neutrophil infiltration, no CD4⁺ and CD8⁺T lymphocytes. In Group B, the inflammatory reaction was observable at different time points, the corneal graft was turbid; and the tissue sections and indirect immunofluorescence staining showed that neutrophil infiltration was predominant, and CD4⁺, CD8⁺T lymphocytes were also seen.

CONCLUSION: Compound betamethasone is able to inhibit the ostrich-rabbit corneal transplantation immune rejection, prolong the survival time of the grafts. The present study lay the foundation for further research and clinical application.]]> 2017/12/18 10:32:18 Yan Cheng, Xian-Ning Liu, Jie Wu, Xiang-Hua Xiao, Shi-Yin Pan and Xiu-Ping Zhu 109true METHODS: Sixty C57BL /6J mice, seven-day-old, were classified into 3 groups: A the normal control group, B the OIR model group, C the ω-3 PUFAs diet group. Each group has twenty mice and separated fed by their lactating mice. The normal control group was fed in a standard atmosphere environment, B, C groups were first fed in a hyper-oxygen atmosphere of(75±2)% oxygen percentage for 5d, then continue fed in a standard atmosphere. The ω-3 PUFAs diet group was fed with dose base on their weight by 7.5mg/kg/d. All mice were sacrificed when they were seventeen-day-old, the relative neovascularization areas(NA)were calculated by fluorescein angiography on flat-mounted retina. The number of endothelial cell nuclei breaking through the inner linmiting membrane(ILM)was counted on hematoxylin and eosin-stained retinal section. The ω-3PUFAs/ω-6PUFAs relative amount and ratio was measured by GC-MS in the retina. A real-time PCR and Western Blot method were used to detect the mRNA, peroxisome proliferator-avtivated receptor–γ(RPAR-γ), vascular endothelial growth factor-A(VEGF-A)and vascular endothelial growth factor receptor 2(VEGFR-2)in the retina.

RESULTS: There was a significant different in all groups on the relative neovascularization areas and the number of endothelial cell nuclei breaking through the ILM(F_NA=20.45, P<0.05; F_ILM=48.66, P<0.05). NA between Group A and B had a significant difference(t=8.64, P<0.05), the same between Group C and B(t=8.91, P<0.05). The cell nuclei breaking through ILM in Group A and B was significantly different(t=38.51, P<0.05), the same in Group C and B(t=19.86, P<0.05). For the relative contain in retina of ω-3PUFAs and ω-6PUFAs, there was a significant different among all groups(F=129.86, F=112.44; all P<0.05). That of Group C was significant different than other two groups(t=23.15, 25.42; t=16.43, 11.95; P<0.05). There were significant different among all groups on ω-3PUFAs/ω-6PUFAs ratio, retinal RPAR-γmRNA expression, retinal VEGF-A mRNA expression and VEGFR-2 mRNA expression(F_ω-3/6=10.30, F_RPAR-γ=138.24, F_VEGF-A=69.12, F_VEGFR-2=52.45; P<0.05). The ω-3PUFAs/ω-6PUFAs ratio of Group C was higher than that of Group B(P<0.05). Compared to Group B, on one hand Group C had a higher expression(P<0.05), on other hand Group C had a lower expression on VEGF-A mRNA and VEGFR-2 mRNA(P<0.05).

CONCLUSION:The diet rich with ω-3 PUFAs uplifts the ω-3PUFAs/ω-6PUFAs ratio and activates RPAR-γ to lower expression of VEGF-A and VEGFR-2 to inhabit oxygen induced retinal neovascularization.]]> 2017/12/18 10:32:18 Qin Li, Zhi Li, Die-Nian Zhang and Shao-Wei Zhang 108true METHODS: A total of 45 patients(45 eyes)with UM who underwent surgical treatment with complete information in our hospital were selected from February 2012 to December 2017. In the same period, 30 patients(30 eyes)who underwent eyeball removal due to ocular trauma and most of the uvea were normal were selected. Real-time quantitative PCR was used to detect the expressions of PAR2 gene in UM and normal choroidal tissues. M23 cells were cultured and divided into PAR2 interference group, negative control sequence group and blank group. Real-time quantitative PCR was used to detect the expression of PAR2 gene in cells. MTT assay was used to detect cell proliferation, and transwell assay was used to detect cell migration and invasion.

RESULTS: The relative expression level of PAR2 mRNA was 1.73±0.13 in UM tissues, and 1.06±0.10 in normal choroid tissues(t=23.732, P<0.001). The relative expression level of PAR2 mRNA in UM tissues was associated with pathological type, scleral invasion, optic disc involvement and extraocular growth(P<0.05). The relative expression level of PAR2 mRNA in PAR2 interference group was lower than that in negative control sequence group and blank group(P<0.05). The absorbance A values at 24h, 48h, 72h and 96h in the PAR2 interference group cells were lower than those in negative control sequence group and blank group(P<0.05). The number of migrated cells and the number of invasive cells in PAR2 interference group were lower than those in negative control sequence group and blank group(P<0.05).

CONCLUSION: PAR2 was highly expressed in UM tissues and was associated with high risk of tumor metastasis. Specific silencing of PAR2 gene expression in M23 cells could effectively inhibit cell proliferation, migration and invasion.]]> 2018/11/19 14:44:07 Mei Ming, Wen Luo and Feng Gao 107true METHODS: An early diabetic retinopathy model was made in rat by intraperitoneal injection of streptozotocin(60mg/kg). The model rats were sacrificed at 4,8,12wk after the establishment of the model, and the eyeballs were removed to make paraffin section and retina sheets were prepared. HE staining was used to detected the retinal morphology and vascularity. Immunohistochemistry(IHC)was employed to examine the expression of Bcl-2, Bax and VEGF in the retina. ADP enzyme staining were conducted to evaluate the retinal vascular morphologic change. Nikon-A1 laser confocal microscopy was used to detect the morphology, fluorescence intensity and distribution of Ca²⁺ in retinal cells.

RESULTS: In the diabetic group, the number of endothelial cells in the inner limiting membrane increased 12wk later. In diabetes mellitus group, there was no vascular area in the middle and peripheral retina, and no vascular area was significantly more than that in the blank control group(P<0.05). There were significant differences in the values of VEGF, Bcl-2 and Bax between diabetic rats and control group(P<0.05). Compared with diabetic rats at 4, 8, 12wk, the fluorescence concentration of calcium ions in RGCs increased gradually, and the ratio of fluorescence staining intensity increased significantly(P<0.05).

CONCLUSION: The expressions of Bcl-2 and Bax were significantly increased, which upgraded the expression of VEGF in the retinas of early diabetic rats. Bcl-2, Bax and VEGF might play an important role in the neovascularization of the retinas in early diabetic rats.]]> 2018/10/22 14:57:10 Li-Jun Song, Yi Wang, Di Chen, Fu-Jun Gao, Hao Ding, Yu-Ming Li and Meng Zhong 106true METHODS:HLEB3 cells cultured in vitro were divided into normal control group(DMEM medium containing 5mmol/L of glucose), oxidative stress group(DMEM medium containing 5mmol/L of glucose and 200μmol/L of hydrogen peroxide), high glucose induction group(DMEM medium containing 30mmol/L of glucose). Apoptosis and cells were detected 24h after culture. Activity and expression of six key glucose metabolic enzymes(fructose-6-phosphate kinase-1, pyruvate kinase, hexokinase, citrate synthase, α-ketoglutarate dehydrogenase and 6-phosphate glucose dehydrogenase)were studied.

RESULTS:Apoptosis of HLEB3 cells was induced by high glucose. The cell viability of high glucose-induced group(63.43%±3.40%)was lower than that of normal control group(100.00%±0.00%)and oxidative stress group(91.90%±5.11%), and the expression levels of six key enzymes of glucose metabolism in high glucose-induced group were lower than that of normal control group and oxidative stress group(all P<0.05).

CONCLUSION:High glucose can induce the expression of glucose-key metabolizing enzymes in HLEB3 cells to decrease, induce apoptosis and affect cell activity.]]> 2019/8/23 9:51:20 Wen-Jing Chen and Ping Liu 105true METHODS: The model of Aspergillus fumigatus keratitis was established and divided into model group and emodin group, 8 in each group and 10 in the normal group. The emodin group was treated with rhubarb, and the model group and the normal group were treated with saline of equal volume. Corneal inflammation index and pathological characteristics were observed. Tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), intercellular adhesion molecule-1(ICAM-1), peroxisome proliferator-activated receptor(PPAR), MAPK and NF-κB protein were detected.

RESULTS: The corneal inflammation index, corneal inflammatory cell count, TNF-α, IL-6, ICAM-1, MAPK and NF-κB protein in emodin group were lower than those in model group, and the expression of PPAR protein was higher than that in model group(P<0.05).

CONCLUSION: Emodin can improve the levels of TNF-α, IL-6 and ICAM-1 inflammatory factors by regulating PPAR, MAPK and NF-κB proteins, and play an anti-inflammatory role in rats with Aspergillus fumigatus keratitis.]]> 2019/8/23 9:51:20 Xiao-Mei Gou, Xue-Li Li, Yuan Sui and Li-Xia Zhou 104true METHODS: Twelve infant rhesus monkeys(aged 2mo)were treated monocularly with PRK(-4.0D)and divided into two groups: AL group(n=6)and NL group(n=6). The monkeys in AL were reared under artificial lighting(indoor).The monkeys in NL were exposed to natural lighting(outdoor)for 4h per day(9:00-11:00 and 15:00-17:00), and to indoor lighting for the rest of the light phase. Corneal haziness after PRK was assessed biomicroscopically using the Fantes scale. At 50d post-PRK, tear fluids of both eyes from 8 monkeys in the two groups(4 animals each group)were collected and analyzed for 11 kinds of cytokines using protein microarray analysis. At 180d post-PRK. The corneas were obtained and evaluated by histopathological examination, immunohistochemistry with antibody to TGF-β1 and α-SMA, and TUNEL. The vitality of SOD and the level of MDA in corneas were detected with WST-1 and lipid peroxidation MDA assay kits, respectively.

RESULTS: The monkeys in AL group exhibited a lesser degree of haze than those in NL group at 40d following PRK(P=0.015). At 50d post-PRK, EGF and TGF-β1 levels in tears were different in PRK-treated eyes between the two groups(P=0.045, 0.038), and TGF-β1 were significantly different between both eyes in the same group(AL: P=0.003; NL: P=0.036). At 180d post-PRK, there were no differences in histological changes, the expression of TGF-β1 and α-SMA, and apoptosis cell staining of the corneal between the two groups. The vatility of SOD and the levels of MDA in corneal epithelium were not different between the two groups(P>0.05).

CONCLUSION: Exposure to natural light in our study could not induce light damage to the normal cornea of the infant rhesus monkeys, but it could aggravate the corneal tissue repair reaction transiently post-PRK.]]> 2019/7/25 14:32:55 Yong Wang, Liang-Ping Liu and Xing-Wu Zhong 103true METHODS: Totally 170 healthy New Zealand white rabbits were randomized into 17 groups, including 15 FARs groups(sodium hydroxymethylglycinate, diazolidinyl urea, imidazolidinyl urea, hydantoin and oxazolidine, each of them has three groups based on dosing concentration: 1/10 maximum allowable concentration group, 1/2 maximum allowable concentration group and maximum allowable concentration group), 1 glutaraldehyde(positive control)group and 1 blank control group. Subconjunctivally injected each FAR in the right eye. 60d after the injection, took the sclera at 1:00 and 7:00 sites of the right eye to make scleral strips. The scleral strip's thickness, elastic modulus, creep rate, ultimate stress and ultimate strain were measured to calculate the scleral biomechanical strength.

RESULTS: Sodium hydroxymethylglycinate, diazolidinyl urea, imidazolidinyl urea, hydantoin and oxazolidine increased the scleral biomechanical strength in a concentration-dependent manner. In these drugs, sodium hydroxymethylglycinate, diazolidinyl urea and oxazolidine had strong cross-linking effects, with obvious effects at the 1/10 maximum allowable concentration.

CONCLUSION: Sodium hydroxymethylglycinate, diazoimidazolidinyl urea, and oxazolidine have strong effects on sclera collagen cross-linking, can significantly improve the biomechanical strength of posterior sclera and have the potential to treat pathological myopia.]]> 2019/7/25 14:32:55 Yin-Cong Xu, Chun-Mei Tong, Ya-Fang Zhao and Chao-Ying Wang 102true METHODS: Apply four kinds of effective components of Qingguang'an and Qingguang'an Chinese medicine suspension to D, E, F, G, H groups after filtration surgery, and pass with group A(blank control group)and group B(model group)Compared with group C(Mitomycin C group), the effects of four effective components of Qingguang'an and Qingguang'an traditional Chinese medicine suspension on collagen fibers, α-SMA and FN in the scar tissue of glaucoma after filtration were observed.

RESULTS: Compare to B group, the ratio of collagen fiber area to E, F, H group, the expressions of α-SMA and the expressions of FN were different(P<0.05).

CONCLUSION: Qingguang'an effective components 2, Qingguang'an effective components 3, mitomycin C and Qingguang'an suspension reduce the proliferation of myofibroblasts and fibroblasts by inhibiting the expression of collagen fibers, α-SMA and FN, and showed obvious anti-glaucoma staining scar after surgery.]]> 2019/5/22 14:15:43 Xue-Si Huang, Jun Peng, Peng-Fei Jiang, Yuan-Bi Li, Juan Yu and Qing-Hua Peng 101true METHODS: We identify third-generation bovine trabecular meshwork cells which were got from tissue mass culture method. Then the relative expression of FN gene and protein in cells were detected by Real-Time PCR and Western-blot after 24h stimulation with 0ng/mL, 0.1ng/mL, 0.5ng/mL IL-6.

RESULTS: The cultured bovine trabecular cells are coincident with what recorded in the book. Real-time PCR and Western blot showed that the amount of FN mRNA produced by cells was 1.000±0.000, 0.213±0.004, 0.056±0.001, 0.019±0.002 respectively, and the protein expression was 1.167±0.012, 0.662±0.009, 0.238±0.011, 0.061±0.011 respectively. There was a significant difference among four groups(P<0.05).

CONCLUSION: Cultured bovine trabecular meshwork cells have a negative correlation with the expression of FN protein after being stimulated by exogenous IL-6, and the results are consistent with the actual state of the disease. We speculate that the IL-6 participate in the pathogenesis and progression of POAG by affecting the expression of FN gene and protein and changing the structure of trabecular meshwork.]]> 2019/5/22 14:15:43 Pan Liu, Jie Meng, Yu-Zhen Liu and Qiang Wang 100true METHODS: Male C57BL/6J mice(free of eye disease)were divided into two groups randomly: control(Ctrl)group and amphotericin B treated(Amph)group, The Ctrl group was given a normal diet, and the Amph group was supplemented with amphotericin B to induce intestinal fungal dysbiosis. After 4wk intervention, corneal epithelial trauma was implemented in both groups. Corneal fluorescein staining was used to evaluate the corneal wound area dynamically. Immunofluorescence staining was applied to observe the changes of corneal epithelial cells and inflammatory cells. HE staining was used to assess the change of corneal thickness.

RESULTS: Compared with Ctrl group, Amph group had delayed re-epithelialization rate and wound repair, less inflammatory cells and thinner corneal.

CONCLUSION: Intestinal fungal dysbiosis delays the corneal wound healing, leading to a weak inflammatory response.]]> 2019/4/22 9:40:42 Lu-Lu Jia, Ding-Li Lu, Su-Su Liu, Yu-Qing Chen, Zhi-Jie Li and Li-Ya Wang 99true METHODS: High glucose was induced into ARPE-19 cells and transfected with miR-152-3p mimics. MTT assay was used to detect cell proliferation activity. Flow cytometry was used to detect apoptosis. Fluorescence quantitative PCR(RT-PCR)was used to detect cells. The expression levels of IGF1 and VEGF in the cells were detected by Western blot and the binding relationship between IGF1 and miR-152-3p was detected by the dual luciferase reporter gene.

RESULTS:High glucose can decrease the activity of ARPE-19 cells, increase the apoptosis rate, inhibit the expression of miR-152-3p and increase the expression of IGF1 and VEGF. Over expression of miR-152-3p can up-regulate high glucose-induced cells. Increased activity and increased apoptosis inhibited the expression of IGF1 and VEGF. The dual luciferase reporter gene assay verified that IGF1 is the target gene of miR-152-3p.

CONCLUSION: miR-152-3p can inhibit the inhibition of high glucose-induced ARPE-19 cell activity and increase apoptosis by targeting IGF1 gene.]]> 2019/4/22 9:40:42 Bo Li, Jin-Peng Chen, Xuan Hu, Shu-Jun Wu and Wen-Bo Qu 98true METHODS: Well-cultured RF/6A cells in vitro were randomly divided into four groups: blank control group, mannitol control group, high glucose group and high glucose+10μg/mL APN group. Cell proliferation, migration and tube formation was detected by CCK-8 assay, transwell chamber and matrigel assay, respectively.

RESULTS: There was no significant difference in the cell proliferation rate(100.00%±0.00% vs 99.09%±0.46%), the number of cell migration(121.60±6.02 vs 119.60±9.39)and the number of tube formation(12.11±0.84 vs 12.22±0.96)between the blank control group and mannitol control group(all P>0.05). The cell proliferation rate of the high glucose group(71.42%±2.29%)was significantly lower than that of the blank control group and the mannitol control group, and the number of cell migration(144.20±9.65)and tube formation(16.00±2.90)of the high glucose group were significantly higher than that of the blank control group and the mannitol control group(all P<0.05). The proliferation rate of cells in the high glucose+APN group(90.87%±1.68%)was significantly lower than that of the blank control group and the mannitol control group, while higher than that in the high glucose group(all P<0.05). The number of cell migration(73.00±6.04)and tube formation(7.89±0.38)in the high glucose+APN group was significantly lower than that of the blank control group, the mannitol control group and the high glucose group(all P<0.05).

CONCLUSION: APN can promote the survival and proliferation of RF/6A cells, and inhibit the migration and tube formation of RF/6A cells under high glucose conditions, suggesting that APN has a protective effect on the damage of RF/6A cells caused by high glucose and inhibits the angiogenesis process under pathological stimulations.]]> 2019/2/21 10:57:17 Rong Li, Guo-Min Yao, Xiao-Di Wang and Yang Yao 97true METHODS: A cultured human RPE cell line was randomly divided into the hypoxia group, 3-methyladenine(3-MA)+ hypoxia group, chloroquine(CQ)+ hypoxia group and the control group. Cells incubated in a hypoxic incubator were set as the hypoxia group. After culture for 24h, western blot was used to analyze the relative expression of autophagy-associated key proteins in each group, including microtubule associated protein 1 light chain 3B(LC3B), Beclin-1 and p62. Then, ELISA was applied to detect the concentration of IL-1β and IL-6 in the cell culture supernatants.

RESULTS: The concentration of IL-1β and IL-6 in the hypoxia group was significantly higher than that in the control group 3-MA+hypoxia group and CQ+ hypoxia group(all P<0.01). The relative expression of LC3B-II/I and Beclin-1 in the hypoxia group was significantly higher than that in the control group and 3-MA+hypoxia group. The relative expression of LC3B-II/I in the hypoxia group was significantly lower than that in the CQ+ hypoxia group(all P<0.01). The relative expression of p62 in the hypoxia group was significantly lower than that in the control group, 3-MA+hypoxia group and CQ+ hypoxia group(all P<0.01).

CONCLUSION: Hypoxia can enhance the expression of LC3B-II/I and Beclin-1 in RPE cells, weaken the expression of p62, and promote the secretion of IL-1β and IL-6. Autophagy inhibitors 3-MA and CQ can reduce the expression of IL-1βand IL-6 in RPE cells.]]> 2019/2/21 10:57:17 Yong-Feng Luo, Rong Li, Qiang Wang and Yang Yao 96true METHODS: The rat retinal Müller cells treated with different concentrations of Aflibercept 100 μL(diluted concentrations of 400, 200, 100 pg/mL, respectively). MTT assay were used to detect cell proliferation, flow cytometry. Apoptosis were detected by the instrument, cell invasion were detected by transwell chamber method, and protein(AKT, STAT3, GAPDH)expression were detected by Western-blot method.

RESULTS: The proliferation activity of Müller cells were decreased with the increased of aflibercept concentration, and compared the difference were statistically significant(P<0.05). After treatment for 48h, the apoptotic rate of Müller cells were gradually increased with the increased of aflibercept concentration, and the invasion and penetration index of Müller cells gradually were decreased, and compared the difference were statistically significant(P<0.05). After 48 h of transfection, the relative expression of AKT protein in Müller cells were not change significantly with the increased of Aflibercept concentration(P>0.05), and the relative expression of STAT3 protein decreased gradually, and compared the difference were statistically significant(P<0.05).

CONCLUSION: Aflibercept can inhibit the expression of STAT3 protein in rat retinal Müller cells, thereby inhibit cell proliferation and invasion and promote apoptosis.]]> 2019/2/21 10:57:18 Qi-Feng Lei and Wei Cai 95true METHODS: Mice were randomly divided into three groups: control group, model group and treatment group. Animal models of retinal oxidative stress injury were established by injecting sodium iodate solution into mice caudal vein. Harvesting the mice retina, Western-blot was used to detect the level of proteins related to DNA damage response such as ATM, NF-κB, cyclin D1 and HMGB1 that associated with DNA repair.

RESULTS: SA-β-gal staining showed that the blue-green deposits in treatment group were reduced than that in model group. The expression of DNA damage reactive protein in treatment group ATM, cyclin D1, NF-κB(0.10±0.009, 0.32±0.01, 0.19±0.002)were significantly lower than those in the model groups(0.77±0.08, 0.70±0.02, 0.36±0.01), and the differences were statistically significant(all P<0.01). At the same time, the expression of DNA repair protein HMGB1 in treatment group(0.927±0.06)were notably higher than that in model group(0.383±0.07)and the difference was statistically significant(P<0.01).

CONCLUSION: H₂ can attenuate senescence by inhibiting oxidative-stress induced DNA damage.]]> 2019/1/17 14:38:59 Rui-Chan Li, Hua Liu and Li-Hua Li 94true METHODS: The rats with hyperglycemia were divided into five groups: model group, Wu Ling San high dose group, low dose group, positive control group and normal groups each group of ten. After oral administration for 12wk, the expression of ICAM-1 and CRP in the serum of rats were measured by ELISA. After HE staining, retinal structure was observed under the light microscope. Blood retinal vascular barrier permeability was measured by Evans blue. The vascular endothelial growth factor(VEGF)expression of retinal tissue were observed by immunohistochemistry

RESULTS: Compared with the normal group, the expression of CRP, ICAM-1 and the EB content in diabetic group were increased. The contents of CRP and ICAM-1 and EB permeability in Wu Ling San high dose group were lower than low dose group and positive control group(all P<0.05). There are retinal ganglion cell layer disorder, retinal edema, and positive VEGF immunohistochemistry expression in the diabetic group. Wu Ling San high dose group can improve retinal structure and reduce retinal edema.

CONCLUSION: Wu Ling San can effectively reduce the expression of inflammatory cytokine, VEGF and retina edema in diabetic retinopathy rats, and also can improve the retinal microvascular in order to protect diabetic retinopathy.]]> 2019/1/17 14:38:59 Xi Chen, Jing Wang and Wei Wei 93true METHODS: HRMECs were cultured and divided into the five groups: normal glucose group, mannitol group,high glucose group, gigantol group and epalrestat group. Cell vitality was detected by the MTS assay, reactive oxygen species(ROS)levels were detected by DCFH-DA fluorescent probe, protein expressions of nuclear factor kappa B(NF-κB)were observed by Western blotting, mRNA levels of AR, hypoxia inducible factor-1α(HIF-1α)and vascular endothelial growth factor(VEGF)were measured by qRT-PCR.

RESULTS: DCFH-DA fluorescent probe levels showed that gigantol reduced ROS generation obviously, and protein expressions of NF-κB, mRNA expressions of AR,HIF-1α and VEGF were all decreased. The expression trends of AR and NF-κB in epalrestat group were consistent with gigantol group.

CONCLUSION: Gigantol and epalrestat could inhibit the activation of AR/NF-κB pathway, and gigantol also could inhibit high glucose induced oxidative stress and in HRMECs, which may have a protective effect on diabetic retinopathy(DR), and improve angiogenesis.]]> 2019/1/17 14:38:59 Mei Zhang, Chun-Xia Li, Fang Wei and Yu Qin 92true in vivo and in vitro, so as to provide the treatment and experimental basis for the treatment of clinical fungal keratitis.

METHODS: Three common pathogenic fungi(Aspergillus flavus, Fusarium Solani and Candida albicans)were used. The experimental group was divided into cross-linking combined natamycin group, natamycin combined riboflavin group, natamycin combined UVA irradiation group, cross-linking group and natamycin group as the control group. The drug was added to the center of the Sabouraud dextrose agar(SDA)plate coated with liquid with each fungal spores with the same maid turbidity of 1.5. Ten minutes later, it was irradiated with collagen cross-linking instrument for 10min and cultured at 28℃ for 36h, and then the inhibition zone size was measured and analyzed statistically. The rabbit model of Fusarium Solani corneal infection was prepared. The model rabbits were randomly divided into model control group, cross-linking treatment group, natamycin treatment group, cross-linking combined natamycin group, 5 rabbits in each group. And another 5 normal rabbits were taken as control, and five rabbits were irradiated in accordance with corneal collagen cross-linking therapy. The results were observed by anterior segment photography, corneal scraping and confocal microscopy, and the ultra micro structural changes of the corneas were observed by electron microscope after the treatment.

RESULTS: Corneal collagen cross-linking alone had shown no effect on each fungus in vitro. Corneal collagen cross-linking combined with natamycin produced significant anti-fungal effect(P<0.05). However, the anti-fungal effect of natamycin combined riboflavin group and natamycin combined ultraviolet light group showed no statistical difference(P>0.05)comparing with the control group. For the model of rabbit fungal infection, the course of disease was about 14d in the natamycin group and CXL combined with natamycin group, and it was about 21d in CXL group. After the treatment, all the groups healed. There were no defects in the corneal epithelium, no mycelium in the corneas, except for more corneal neovascularization. The results of the anterior segment photography showed that the treatment effect of the cross-linking combined natamycin group was better than other groups, with fewer scar tissue, better corneal healing and relatively short course of disease.

CONCLUSION: Corneal collagen cross-linking combined with natamycin treatment is able to enhance anti-fungal effect, promote corneal healing, and shorten the course of disease. So it is a promising therapeutic technique for the clinical treatment of fungal keratitis.]]> 2018/12/17 9:54:34 Shi-Yin Pan, Na An, Xiang-Hua Xiao, Yan Cheng, Yang-Zheng Wang, Xian-Ning Liu, Xiu-Ping Zhu and Jie Wu 91true METHODS: A total of 160 postnatal day(P)7 C57BL/6 mice were obtained. All mice except normal control group were exposed to(75±2)% oxygen environment for 5d and then kept in room air for another 5d to establish the OIR mice model. All mice in normal control group(40 mice)were exposed to room air only. At P12 and P14, respectively, mice in PEDF treatment group were injected intravitreously with recombinant human PEDF(2μg/eye,1μL)in the right eye, while mice in treatment control group were injected intravitreously with the same volume of vehicle [1μL, 10mmol/L phosphate buffered saline(PH7.4), PBS] in the right eye. All mice were euthanized at P17. Eyes were whole mounted and stained with Lectin to observe the growth of abnormal RNV; And retinal specimens were prepared for PEDF, MCP-1 protein and mRNA analysis by Western blot and real time RT-PCR respectively.

RESULTS: Changes of retinal vessels had been detected by fluorescence microscopy on flat-mounted retina. The relative RNV areas were significantly increased in OIR model group compared with those in normal control group(P<0.01). However, the relative RNV areas were significantly reduced in PEDF treatment group compared with those in PBS treatment control group(P<0.01). The specific expression of MCP-1 protein and mRNA in the OIR model group were higher than those of normal control group, presenting a statistically significance(both P<0.05). The specific expression of PEDF protein and mRNA in the OIR model group showed a considerable decline in comparison with normal control group, presenting a statistically significance(both P<0.01). And the specific expression of MCP-1 protein and mRNA in those of PEDF-treated group showed a considerable decline in comparison with PBS-treated group, and the differences were statistically significant(both P<0.05). However, there were increase of the expression of MCP-1 protein and mRNA between normal control group and PEDF-treated group, presenting no statistically significance(both P>0.05).

CONCLUSION: PEDF could inhibit oxygen-induced retinal neovascularization and down-regulate retinal MCP-1 expression under hypoxia, which may underlie its anti-neovascularization effects and play a role of protection in ischemic retinopathy.]]> 2018/12/17 9:54:34 Ya-Nuo Wang and Lei Zhang 90true METHODS: The rats were randomly divided into blank control group, model group, low dose DHA group(L-DHA group), medium dose DHA group(M-DHA group)and high dose DHA group(H-DHA group). The rat model of dry ARMD was established by light injury. He staining was used to observe the pathological changes of retina, TUNEL method to detect the apoptosis of retinal cells, transmission electron microscopy to observe the ultrastructure of retinal ganglion cells, enzyme-linked immunosorbent assay to detect the levels of TNF - α and IL-6 in retina, Western blot to detect the expression of p-PI3K, p-Akt, Bax, Bcl-2, p-NF-κBp65 and cleved-caspase-3 protein in retina.

RESULTS: Compared with the blank control group, the total thickness of retina, the thickness of outer nuclear layer and inner nuclear layer, the expression of p-PI3K, p-Akt and Bcl-2 protein in the retina of the model group decreased(P<0.05), the apoptosis index in the ganglion cell layer and outer nuclear layer, the level of TNF-α and IL-6 in the retina, the expression of Bax, p-NF-κBp65 and cleared caspase-3 protein increased(P<0.05). Compared with the model group, the total thickness of retina, the thickness of outer nuclear layer and inner nuclear layer, the expression of p-PI3K, p-Akt and Bcl-2 protein in retina increased in M-DHA group and H-DHA group(P<0.05), the apoptosis index in ganglion cell layer and outer nuclear layer, TNF-α and IL-6 levels in retina, Bax, p-NF-κBp65 and cleved caspase-3 protein expression decreased(P<0.05).

CONCLUSION: DHA may inhibit photoreceptor apoptosis by activating PI3K /Akt pathway.]]> 2019/11/21 15:09:55 Hui Wang, Ling Shen and Xiang Ji 89true METHODS: Totally 40 C57BL/6 male mice were randomly divided into the normal group, diabetes group, siRNA-HIF-1α group and siRNA-NC group. The diabetic models were constructed. The histopathological change of the retina of the mice was observed in each group. The microvessel density(MVD)was detected by immunohistochemistry. The expressions of HIF-1α, VEGF,NF-κB, IL-1, IL-6 and TNF-α mRNA in the retina in each group were detected by real-time quantitative PCR. The expressions of HIF-1α, ET-1 and vWF proteins in the retina in each group were detected by Western blot.

RESULTS: The body weights of diabetes group, siRNA-NC group and siRNA-HIF-1α group were lower than the normal group, while the blood glucose levels were higher than the normal group(All P<0.05). The MVD in the diabetic group and siRNA-NC group were significantly higher than those in the normal group, while the siRNA-HIF-1α group were significantly lower than the diabetes group and siRNA-NC group(All P<0.05). Compared with the diabetes group and siRNA-NC group, the relative expression levels of HIF-1α, VEGF, NF-κB, IL-1, IL-6 and TNF-α mRNA in the retina in the siRNA-HIF-1α group were decreased(All P<0.05). Compared with the diabetes group and siRNA-NC group, the relative expression levels of HIF-1α,ET-1 and vWF proteins in the retina in the siRNA-HIF-1α group were decreased(All P<0.05).

CONCLUSION: Specific silencing of HIF-1α gene could protect the retina of DR. The mechanism may be related to the reduction of angiogenesis and vascular endothelial injury.]]> 2019/11/21 15:09:55 Rui-Ping Zhang and Hong-Ling Liu 88true METHODS: Thirty-six adult male rabbits were divided into two groups. Group A was treated with false castration, group B was pseudo-implantation group, and group C was implanted with lacrimal canal. Fluorescein(FL)staining and scoring and SⅠt examination, and confocal microscopy were performed at 2, 4, 6wk after implantation.

RESULTS: The cornea of group A was smooth and clear, and the FL staining was not obvious. In group B, the cornea was rough, and the corneal epithelium was spotted with stained spots. The corneal surface of group C was smoother and more transparent than that of sham operation group and the punctate staining was not obviously. The FL staining scores and SⅠt examination were significant(P<0.05)at 2, 4, and 6wk after implantation. FL staining and scoring and SⅠt, tear protein determination, confocal microscopy changes in corneal nerve. Confocal microscopy showed that group A was a relatively straight plexus, the amount was large, clearly visible. In the group B, the inferior nerve of the corneal epithelial cells was visible and the number was reduced. The corneal epithelial nerve in group C was relatively straight, and the number was slightly reduced compared with the normal male rabbit corneal epithelial nerve.

CONCLUSION: The amniotic lacrimal duct stent can significantly improve the symptoms and signs of castrated castrated male rabbit dry eye syndrome, and has a therapeutic effect.]]> 2019/10/23 15:17:25 Biao Li, Qing Yuan, Pei-Wen Zhu, Qi Lin, You-Lan Min, Wen-Qing Shi, Lei Ye and Yi Shao 87true METHODS: Immunohistochemistry and Western-blot assays were used to detect the expression levels of TGFBI and LC3 in corneal dystrophy and normal corneal tissues. TGFBI overexpression vector was transfected into corneal stromal fibroblasts, and then the cells were treated with 5, 10, 20, 40mmol/L LiCl for different times(0, 1, 6, 12h), and Western-blot assay was performed to evaluate the expression levels of TGFBI and LC3, and CCK-8 assay was carried out to assess cell proliferation activity.

RESULTS: TGFBI and LC3 were highly expressed in corneal tissues of patients with corneal dystrophy. TGFBI overexpression inhibited the proliferation ability of corneal stromal fibroblasts(P<0.05). LiCl inhibited the expression levels of TGFBI and LC3, and enhanced the cell proliferation activity in corneal stromal fibroblasts transfected with mutant TGFBI(P<0.05).

CONCLUSION: LiCl promoted the proliferation and autophagy of corneal stromal fibroblasts, and its mechanism may be related to down regulated expressions of TGFBI and LC3.]]> 2019/10/23 15:17:25 Dan-Yao Nie, Ming Li, Lin Ye, Wen-Ling He and Xin-Hua Liu 86true METHODS: C57BL/6J mice were randomly divided into the control group and diabetic retinopathy(DR)model group. The cultured rhesus monkey choroido-retinal endothelial cells(RF/6A cells)were randomly divided into the control group(cultured in low-glucose environment)and the high-glucose group(cultured in medium with 25mmol/L D-glucose), and the AGGF1 protein expression in the cells was detected by immunofluorescence assay. RF/6A cells were then divided into the control group and AGGF1 treatment group, and cell proliferation, migration and tube formation was detected by CCK-8, Transwell and Matrigel, respectively.

RESULTS: AGGF1 protein was expressed in all layers of the retinas and in vascular endothelial cells. The expression of AGGF1 in the retinas of DR group(0.17±0.05)was significantly higher than that of the control group(0.07±0.02)(P<0.05). AGGF1 protein was expressed in RF/6A cells in both the high glucose group and the control group, and the expression of AGGF1 in RF/6A cells under high glucose was significantly higher(0.63±0.10)than that in the control group(0.40±0.03)(P<0.05). After 12h of treatment, the cell proliferation rate(114.88%±0.84%)in the AGGF1 group was significantly higher than that in the control group(100.00%±2.17%)(P<0.05). After 24h of treatment, the cell proliferation rate of the AGGF1 group(157.35%±1.89%)was significantly higher than that of the control group(142.77%±0.50%)(P<0.05). After 48h of treatment, the cell proliferation rate of the AGGF1 group(185.39%±1.90%)was significantly higher than that of the control group(160.17%±1.33%)(P<0.05). After 12h of treatment, the number of migrated cells(127.00±7.00)in the AGGF1 group was significantly higher than that in the control group(90.33±6.66)(P<0.05). After 12h of treatment, the number of tube formation(33.67±1.15)in the AGGF1 group was significantly higher than that in the control group(15.33±3.51)(P<0.05). The total tube length in AGGF1 group(8226.33±288.55)μm was significantly higher than that in the control group(6463.33±938.01)μm(P<0.05).

CONCLUSION: The expression of AGGF1 protein was significantly increased in diabetic retinal tissues and retinal vascular endothelial cells induced by high glucose. AGGF1 can promote the proliferation, migration and tube formation of retinal vascular endothelial cells, suggesting that AGGF1 may be involved in retinal neovascularization of DR.]]> 2020/8/19 19:18:37 Rong Li, Guo-Min Yao, Hong-Lin Yan, Li-Wang, Tian-Zhi Cai, Bo-Bo Yuan, Wei Ju and Li-Na Wang 85true METHODS: The experiment was divided into 6 groups, blank control group and experimental group, that is, five different concentrations of d-δ-tocopherol(40, 60, 80, 100, 120)μmoL/L. The proliferation inhibition rate of each group was detected by thiazolam(MTT)assay. The morphology of human lens epithelial SRA cells was observed under inverted microscope. Cell cycle was detected by flow cytometry and the expression of bcl-2, bax, Cyclin D1, P21 protein was detected by Western Blot(WB).

RESULTS: With the increase of d-δ-tocopherol concentration, the SRA cells decreased significantly compared with the control group; the MTT results showed that with the increase of d-δ-tocopherol concentration, the inhibition rate of cell proliferation increased gradually, the difference was statistically significant(P<0.05); cell cycle: with the increase of the concentration of tocopherol drugs in the experimental group, the proportion of cells in the S phase increased gradually compared with the control group, the cells were blocked in the S phase, the difference was statistically significant(P<0.05); Western blotting: after 48h of d-δ-tocopherol intervention human lens epithelial SRA cells, the P21, Cyclin D1 and bcl-2 expression of human lens epithelial cells gradually decreased, and the expression of bax gradually increased, which was statistically significant(P<0.01).

CONCLUSION: d-δ-tocopherol can significantly inhibit the proliferation of human lens epithelial SRA cells and block the cell cycle in S phase. d-δ-tocopherol can inhibit the proliferation of human lens epithelial cells. The proliferation of human lens epithelial SRA cells may be achieved by inhibiting the expression of bcl-2, P21, Cyclin D1 and inducing the expression of bax.]]> 2020/8/19 19:18:37 Cheng Qin, Xin-Sheng Zeng, Bo Peng, Yu-Ni Tang, You-Di Zhou and Bo Song 84true METHODS: ARPE-19 cells cultured in vitro were randomly divided into four groups, which were control group, 447nm blue light group, 456nm blue light group and 468nm blue light group. The cells in control group were cultured under normal conditions whereas the cells in blue light group were irradiated with different wavelengths of OLED blue light with the illumination intensity of 200Lux for 72h. Live/Dead staining assay, CCK-8 assay and real-time PCR were performed to compare the effects of different wavelengths of blue light on the morphology, cell viability, proliferation capacity, mRNA expression level of visual cycle biomarkers and inflammatory biomarkers of ARPE-19 cells, respectively.

RESULTS: After blue light irradiation, the abnormal morphology and the decrease of cell confluence of ARPE-19 cells were observed. Furthermore, with the decrease in the wavelength of blue light, the inhibition effect of blue light on RPE proliferation was enhanced, and the mRNA expression of the proliferation marker Ki-67 and visual cycle biomarkers LRAT, CRALBP, RDH and IRBP decreased. In addition, the mRNA expression levels of inflammatory factors MCP-1 and IL-6 in RPE cells were up-regulated with the decrease in the wavelength of blue light.

CONCLUSION: Our data demonstrated that blue light in different wavelengths exerted detrimental effects on RPE cells. The shorter the wavelength of blue light was, the more severe damage it caused on the RPE cells.]]> 2020/7/22 11:15:57 *, Zhi-Min Tang^*, Yu-Yao Wang, Xiao-Chan Dai, Min Luo and Ping Gu]]> Ya-Han Ju^*, Zhi-Min Tang^*, Yu-Yao Wang, Xiao-Chan Dai, Min Luo and Ping Gu 83true METHODS:In vitro cultured RF/6A cells were randomly divided into the control group, hypoxic injury(induced by CoCl₂ stimulation)group and hypoxic injury + adiponectin(5μmol/L, 50μmol/L and 100μmol/L)group. Cell viability was assessed using the MTT assay and optimal concentration of adiponectin was selected. Western blot was used to detect the expression of Bax and Bcl-2 in RF/6A cells. Reactive oxygen species(ROS)detection kit was used to detect the content of ROS in RF/6A cells.

RESULTS: Compared with the control group, the cell viability of RF/6A cells in the hypoxic injury group and each adiponectin pretreatment group decreased(all P<0.01). Compared with the hypoxic injury group, the cell viability of RF/6A cells in each adiponectin pretreatment group was significantly increased(all P<0.05), and adiponectin of 50μmol/L was the appropriate protective concentration. Compared with the control group, the viability of RF/6A cells decreased, the protein expression level of Bax increased, the protein expression level of Bcl-2 decreased, and the content of ROS increased in the hypoxic injury group(all P<0.01). Compared with the hypoxic injury group, the viability RF/6A cells increased, the expression level of Bax decreased, the expression level of Bcl-2 increased, and the content of ROS decreased in the adiponectin pretreatment group(all P<0.01).

CONCLUSION: Our findings suggest that adiponectin can significantly alleviate retinal vascular endothelial cell damage and apoptosis caused by hypoxia, and the mechanism may be related to the inhibition of oxidative stress by adiponectin.]]> 2020/7/22 11:15:57 Guo-Min Yao, Gulinuer Maimaiti, Rong Li, Ying Deng and Cong Jiao 82true METHODS:Thirty healthy 8-week-old male Brown Norway rats were randomly assigned into three groups equally. The left eye of group A was blank group, group B was the left lacrimal gland extirpation model, the left tear gland of group C was injected with botulinum toxin A. We compared the data of Schirmer I test, tear break-up time(BUT), and the corneal fluoresceince staining scores at different times(1d before experiment, 3d, 7d, 14d, 28d, and 42d after the surgical process). We observed pathological changes of conjunctiva, cornea and lacrimal gland at 42d, and we used real-time polymerase chain reaction to analyze interleukin-6(IL-6), tumor necrosis factor-α(TNF-α)and epithelial growth factor(EGF).

RESULTS:At the 3d, compared with group A, the tear secretion of both group B and group C were continuous decrease(P<0.05). At the 7d, compared with group A, the BUT of both group B and group C began to decreased(P<0.05), and the corneal epithelial staining scores of both group B and group C began to significantly increase(P<0.05). There was no statistical difference in the above clinical data between group B and group C(P>0.05). The corneal epithelial cells in group A was set as normal morphology, while the corneal epithelial cells in group B and group C showed filamentous separation of surface cells to varying degrees, and the number of conjunctival goblet cells was decreased. The lacrimal gland of group C was obviously atrophic. In conjunctival and corneal tissues, the expression of EGF, TNF-α and IL-6 were significantly increased in group B and group C, which was statistically significant compared with group A(P<0.05). The expression of EGF and TNF-α didn't altered significantly between group B and group C(P>0.05), however, the expression of IL-6 in group B was much higher than that in group C(P<0.05).

CONCLUSION:In this study, we proved that both lacrimal gland extirpation and lacrimal gland injection botulinum toxin A could construct a stable aqueous tear deficiency dry eye rat model. The appropriate animal model should be selected according to the experimental design and research purpose.]]> 2020/7/22 11:15:57 Zi-Yi Dong, Ming Ying, Xi Yang and Yi Ma 81true METHODS: SD rat retinal vascular endothelial cells(RRVEC)were cultured and the RRVEC was divided into control group(NG)and high glucose group(HG). The high glucose-induced RRVECs were harvested separately or co-transfected with miR-96-5p mimic, miR-NC, si-FOXO4, si-NC. The expression of miR-96-5p and FOXO4 was detected by qRT-PCR and Western blotting, respectively. MTT assay was used to detect the proliferation activity. Flow cytometry was used to detect the apoptosis rate. The dual luciferase reporter assay validated the target gene of miR-96-5p. Western blotting was used to detect the expression of CyclinD1, p21, p27, Bcl-2, Bax and cleaved-caspased-3.

RESULTS:The expression levels of miR-96-5p, CyclinD1 and Bcl-2 in RRVEC were significantly decreased after high glucose treatment, and the expression levels of FOXO4, p21, p27, Bax and cleaved-caspased-3 were significantly increased, inhibiting cell proliferation activity, but promoting apoptosis. Overexpression of miR-96-5p and inhibition of FOXO4 expression increased the expression levels of CyclinD1 and Bcl-2, inhibited the expression of p21, p27, Bax, cleaved-caspased-3, enhanced cell proliferation and inhibited apoptosis. Dual luciferase reporter assay demonstrated that FOXO4 was a target gene for miR-96-5p. Overexpression of FOXO4 reversed the effect of miR-96-5p overexpression on high glucose-induced proliferation and apoptosis of RRVEC.

CONCLUSION:miR-96-5p inhibits high glucose-induced apoptosis of rat retinal vascular endothelial cells and promotes cell proliferation by targeting FOXO4.]]> 2020/7/22 11:15:57 Huan Li and Lu Lu 80true METHODS: Kunming mice were used as research objects, which were divided into control group, model group and quercetin group. Fundus examination was showed whether yellow-white like glassy sputum substances appeared in the fundus of each group of mice; OCT was used to examine the retinal thickness of each group of mice; HE staining was used to observe the changes of retinal morphology in each group of mice; FFA was observed the fundus vascular integrity of each group of mice. The activities of SOD, GSH-Px, CAT and the contents of ROS and MDA in serum were detected by ELISA; Western blot was used to detect the expression of Nrf2/Keap1/ARE related proteins in the retina of each group.

RESULTS: Quercetin can reduce the yellow and white glassy wart substance in the fundus of mice and increase the thickness of the retina(P<0.05), and the points of retinal vascular leakage is significantly reduced. Compared with the model group, the a-wave amplitude and b-wave amplitude of the quercetin group were significantly higher than those of the model group(P<0.01); Quercetin can make the retinal structure of mice clearer, necrosis and shed part of the outer nuclear layer, and reduce the content of ROS and MDA in mouse serum(all P<0.05), and increase the activities of SOD, GSH-Px, and CAT(all P<0.05). Compared with the model group, the expression of Nrf2 protein in the retinal cytoplasm of mice in the quercetin group was up-regulated(P<0.05), and the expression of Nrf2 protein in the nucleus was down-regulated(P<0.05), GCL protein expression was down-regulated(P<0.05).

CONCLUSION: Quercetin improved the oxidative stress state after retinal photodamage through the Nrf2/Keap1/ARE pathway, protected the retinal function, and protected against age-related macular degeneration.]]> 2020/6/22 14:51:53 Ting-Yan Chen and Yang Zhou 79true METHODS: A total of 46 paraffin-embedded specimens of common lacrimal gland epithelial tumors and other lacrimal gland tumor types, including 20 cases of lacrimal gland pleomorphic adenoma, 12 cases of pleomorphic adenocarcinoma, and 14 cases of adenoid cystic carcinoma, as well as ten normal lacrimal glands were analyzed for the expression of EGFL7 protein. For all specimens, the tumor microvascular networks were also labeled with anti-CD34 antibody and the tumor MVD was calculated. The proliferative activity of tumor cells containing Ki67.

RESULTS: EGFL7 protein was scored as positive with the presence of brown color in the cytoplasm, and was mainly observed in cells of lacrimal gland pleomorphic adenocarcinomas and adenoid cystic carcinomas. Immunohistochemical staining showed that EGFL7 was not expressed in normal lacrimal gland tissue. The rates of expression of EGFL7 in lacrimal gland pleomorphic adenomas, lacrimal gland pleomorphic adenocarcinomas, and lacrimal adenoid cystic carcinomas were 5%(1/20), 83%(10/12), and 86%(12/14), respectively. The EGFL7 expression in both malignant tumor types was significantly higher than that in pleomorphic adenomas and normal lacrimal gland tissues(P<0.001). CD34 staining colored the tumor microvascular network brown-yellow in single cells or clustered cell populations. The expression of CD34 in lacrimal gland pleomorphic adenocarcinomas(32.58±14.46)and adenoid cystic carcinomas(43.43±4.60)was significantly higher than that in pleomorphic adenomas(4.20±1.19)(P<0.001). Ki67 staining appeared as a brownish color in cell nuclei, indicating proliferative activity. The expression of Ki67 in lacrimal gland pleomorphic adenocarcinomas(44.83±13.68)and adenoid cystic carcinomas(26.29±8.44)was significantly higher than that in pleomorphic adenomas(2.80±3.14)and normal tissues(0.40±0.70)(P<0.001). Furthermore, the expression of EGFL7 protein was positively correlated with high MVD and Ki67 expression in lacrimal epithelial tumors(r_s=0.897, P<0.001; r_s=0.837, P<0.001).

CONCLUSION: The correlation of EGFL7 expression with high MVD and Ki67 expression suggests that high EGFL7 expression plays an important role in promoting tumor angiogenesis and tumor proliferation.]]> 2020/6/22 14:51:53 Qi-Qi Xing, Fang Zhang, Dong Liu and Fu-Ling Liu 78true METHODS: In the research, the retinal Müller cells are regarded as research subjects, and the control group, hypoxia group and experiment group are set up. The cells of control group are cultivated in normal environment. The cells of hypoxia group are cultivated in hypoxia environment. The cells of experiment group are cultivated in hypoxia environment and are treated with the Apelin-13(1μmol/L). MTT method is used to monitor the changing of the cell viability, and the crystal violet staining method is adopted to observe the cell morphology. In addition, the immunofluorescence staining method is used to test the expression of GFAP and YAP and the TUNEL staining method is used to monitor the cell apoptosis situation and the apoptosis index is calculated. The protein staining method is used to observe the changing of the expression of p-LATS1, p-YAP, LATS1 and YAP protein.

RESULTS:The separated and extracted Müller cells grow on the wall and show elongation, polygon and circular shapes. The cytoplasm is plentiful and the cell nucleus show circular shape. The GFAP expression of the cell is positive. The treatment with 0.1, 1, 10μmol/L Apelin-13 can obviously prevent the Müller cell viability decreasing induced by hypoxia(P<0.05 or P<0.01). Compared with the control group, the cell apoptosis index of hypoxia group is obviously increased(P<0.01). However, compared with the hypoxia group, the cell apoptosis index of experiment group is obviously decreased(P<0.01). The p-LATS1 and p-YAP protein expression of the control group and hypoxia group does not have big difference. Compared with hypoxia group, the p-LATS1 and p-YAP protein expression of experiment group is obviously decreased(P<0.01). The YAP protein expression of cell nucleus of control group and hypoxia group does not have great difference. Compared with hypoxia group, the cell nucleus expression of YAP cell is gretaly increased(P<0.01).

CONCLUSION: Apelin-13 can be used to prevent the retinal Müller cells apoptosis caused by the hypoxia, which may be related to the regulation of YAP into the nucleus.]]> 2020/5/25 15:42:58 Lei Sun and Yong Tao 77true METHODS: BRVO model rats were induced by photochemistry induction and randomly divided into BRVO model group and TA(1, 7, 21)d groups; at the same time, blank control group was set for comparison. The intraocular pressure of rats was measured by ophthalmotonometer; the condition of rat fundus was observed fluorescein fundus color photography(FFA)and optical coherence tomography(OCT); retinal angiogenesis related factors vascular endothelial growth factor(VEGF)and vascular endothelial growth factor receptor 2(VEGFR2), the protein expressions of Notch pathway important factors Notch 1, Jagged 1 and DLL4 were detected in rat retina by Western blotting(WB).

RESULTS: In the normal control group, the fundus vessels were arranged neatly and in a clear state. In the BRVO model group, edema appeared in the fundus, the retina turned white, the arrangement of blood vessels was disordered, the optic disc pit was disappeared, retinal vessels were in the state of vasoconstriction. In TA 1, 7 and 21d groups, edema gradually decreased, blood vessels expansion and bending gradually slowed down, and the optic disc pit was restored. Compared with the blank control group, the intraocular pressure of BRVO model group increased, the thickness of the retina increased at the injured site and 250μm far from injured site, the protein expressions of VEGF, VEGFR2, Notch1 and Jagged1 increased, the protein expression of DLL4 protein was decreased(P<0.05). Compared with the BRVO model group, in TA 1d group, the retinal thickness decreased at 250μm far from injured site, the protein expressions of VEGFR2, Notch1 and Jagged1 decreased, the protein expression of DLL4 protein increased; in TA 7d group, the retinal thickness was decreased at the injured site and 250μm far from injured site, the protein expressions of VEGFR2, Notch1 and Jagged1 decreased, the protein expression of DLL4 protein increased; the intraocular pressure of TA 21d group decreased, the thickness of the retina decreased at the injured site and 250μm far from injured site, the protein expressions of VEGF, VEGF R2, Notch1 and Jagged1 decreased, the protein expression of DLL4 protein increased(P<0.05).

CONCLUSION: Vitreous injection of TA may inhibit angiogenesis by regulating Notch pathway to inhibit the activation of VEGF, thus achieving the retinal protection in BRVO rats.]]> 2020/5/25 15:42:58 Sha-Sha Han, He-Peng Zhang and Yue-Feng Li 76true METHODS: In order to investigate the changes of EMT and autophagy induced by high glucose, HLE-B3 cells were divided into two groups. In NC group, cells were cultured in DMEM with 5.5mmol/L glucose, and in HG group, cells were treated with DMEM in addition with 30mmol/L glucose for 12h, 24h, and 48h. Western blot was used to detect the expression of EMT-marker proteins(E-cadherin and α-SMA)and autophagy-marker proteins(LC3, Beclin 1 and SQSTM1/p62). Wound healing assay was conducted to observe the migration ability. To investigate the regulation of autophagy on EMT, we employed rapamycin, an agonist of autophagy. HLE-B3 cells were divided into 4 groups. Two of them were mentioned as above, and the other two groups were treated with high glucose combined with DMSO(DMSO)and high glucose combined with 200nmol/L rapamycin(RAPA), respectively. Migration ability of cells was evaluated by Transwell assay. Expressions of proteins, such as EMT marker proteins, molecules in TGF-β signaling pathway(TGF-β2, Smad2/3, p-Smad2/3, Snail), and autophagy markers were detected by Western blot. The intracellular co-localization of SQSTM1/p62 and Smad2/3 was observed by immunofluorescence staining, and their interaction was confirmed by co-immunoprecipitation assay.

RESULTS: The expression of E-cadherin, LC3 Ⅱ/Ⅰ, and Beclin 1 in HLE-B3 cells of HG group gradually decreased(F=67.52, 163, 206; all P<0.0001), the expressions of α-SMA, SQSTM1/p62 increased with time(F=53.37, 302.1; all P<0.0001), and cell migration also increased compared with the cells in NC group(all P<0.001), indicating that high glucose stimulated EMT and suppressed autophagy. After treatment with rapamycin, the expressions of LC3 Ⅱ/Ⅰ and E-cadherin increased, the expressions of α-SMA, p-Smad2/Smad2, p-Smad3/Smad3 and Snail decreased(all P<0.05), and the expressions of TGF-β2 did not change(all P>0.05)in RAPA group compared with HG group and DMSO group, cell migration was also suppressed(all P<0.001), indicating that Rapamycin down regulated the expressions of molecules in TGF-βsignaling pathway after activation of autophagy, which resulted in inhibiting EMT. Immunofluorescence staining showed co-localization of SQSTM1/p62 and Smad2/3 in cytoplasm. Co-immunoprecipitation confirmed the combination between SQSTM1/p62 and Smad2/3.

CONCLUSION: High glucose stimulates the process of EMT and suppresses the autophagy in HLE-B3 cells. Autophagy regulates EMT by interacting with Smad2/3 via SQSTM1/p62, altering the amount of Smad2/3 which works in the TGF-β signaling pathway.]]> 2020/4/26 11:24:08 Ji-Yuan Ma, Wei Ye, Ji Li, Rui Pei, Meng-Mei He, Jing-Bo Su, Dong-Jie Sun, Qi-Wu Zhou and Jian Zhou 75true METHODS: MicroRNA-34a expression levels in ARC lens and transparent lens epithelial cells were detected by qRT-PCR. MicroRNA-34a mimics, MicroRNA-34a inhibitors and empty liposome(control group)were transfected into SRA01/04 cells by liposome transfection kit. Annexin V-FITC/PI double staining was used to detect the effect of MicroRNA-34a on the apoptosis of human lens cell line SRA01/04. The expression of Cdc42 and Rac1 protein was detected by western blot.

RESULTS: The expression level of MicroRNA-34a in anterior capsular tissue of transparent lens was significantly lower than that in ARC anterior capsular tissue(P<0.05). The positive rates of SA-β-gal in the MicroRNA-34a mimics group, the control group and the MicroRNA-34a inhibitors group were(87.56±2.34)%,(12.22±2.74)% and(3.45±0.45)%, respectively. The positive rates of SA-β-gal in the MicroRNA-34a mimics group was significantly higher than the control group, while the SA-β-gal positive rate in the MicroRNA-34a inhibitors group was significantly lower than that in the control group(P<0.05). The apoptosis rate of the MicroRNA-34a inhibitors group, control group and MicroRNA-34a mimics group were(5.87±1.22)%,(12.26±2.14)% and(29.45±3.12)%, respectively. The apoptosis rate of the MicroRNA-34a mimics group was significantly higher than that of the control group, while that of the MicroRNA-34a inhibitors group was significantly lower than that of the control group(P<0.05). The expressions of Cdc42 and Rac1 in the MicroRNA-34a mimics group were significantly higher than those in the control group(P<0.05), while the expressions of Cdc42 and Rac1 in the MicroRNA-34a inhibitors group were significantly lower than those in the control group(P<0.05).

CONCLUSION: MicroRNA-34a may promote the senescence and apoptosis of human lens epithelial cells by up-regulating Cdc42 and Rac1.]]> 2020/4/26 11:24:08 Wen Lan, Zhi-Jun Chen and Lu Zhou 74true METHODS: The cells in logarithmic growth stage were divided into control group(glucose concentration 5.5mmol/L), low glucose group(glucose concentration 5mmol/L), high glucose group(glucose concentration 30mmol/L). CXCL9(100ng/mL), CXCL10(10ng/mL)and CXCL11(100ng/mL)were added respectively, cultured for 24, 48 and 72h. CCK-8 method was used to detect cell proliferation, RT-PCR was used to detect CXCR3 mRNA expression, and immunofluorescence was used to detect Ki-67 expression.

RESULTS: The results of CCK-8 method showed that the absorbance value of the control group increased gradually with the increase of time after adding three exogenous chemokines. The absorbance value of the low glucose group increased first and then decreased, reaching the peak at 48h. The absorbance value of the high glucose group decreased generally. The results of RT-PCR showed that the expression of CXCR3 mRNA in low glucose group and high glucose group was higher than that in 24h after adding CXCL9, CXCL10 and CXCL11 for 48 and 72h. The results of immunofluorescence showed that the fluorescence intensity of Ki-67 decreased in the low and high glucose 72h after adding CXCL9, CXCL10 and CXCL11. The change in the high glucose group is more obvious.

CONCLUSION: Exogenous CXCL9, CXCL10 and CXCL11 can decrease the activity of human umbilical vein cell under high glucose environments and induce the increase in CXCR3 expression. The increase of CXCR3 reached the highest after adding exogenous CXCL10 and CXCL11, suggesting a target for clinical intervention of diabetic retinopathy.]]> 2020/4/26 11:24:08 Hai-Lin Hu and Min Luo 73true METHODS: Constructing a bilayer lens by adhering together two corneal stromal lenses with human fibrin sealant(FS). Human corneal fibroblasts were isolated and cultured from Smile derived corneal stromal lenses in vitro, and the toxicity of FS on human corneal fibroblasts was detected by MTT method. The bilayer lenses were then placed in anhydrous glycerin, sodium hyaluronate eye drops, a simulated wet room environment and fetal bovine serum groups respectively, and stored at 4℃ for 14d. The transparency, hardness and stability of the scaffolds were then compared. Afterwards, the bilayer lens scaffolds were stored in anhydrous glycerin at room temperature, 4℃ and -20℃. After 14d of preservation, the diverse effects of temperature on the transparency and hardness of the scaffolds were compared.

RESULTS: MTT results showed that the cells of the experimental group and the control group had similar proliferation trend within 0-72h. The cytotoxicity rating of the experimental group was 0 at 36-48h and 1 at 24h and 60-72h. The relative survival rate of the cells within 0-72h was over 90%. FS-bonded bilayer lens scaffold had a smooth surface, close bonding, good transparency and suitable hardness. After 14d of storage at 4℃, none of the nine bilayer lens scaffolds in the anhydrous glycerol group showed signs of cracking cracking after rehydration, and their transparency was good. In the sodium hyaluronate group, three of the nine scaffolds cracked and the remaining six were still intact. In the simulated wet room environment group, none of the 9 scaffolds cracked, but there were different degrees of shrinkage, their surface was rough and transparency was lower. In the fetal bovine serum group, all the 9 stents were cracked, and the single corneal stromal lens was soft and edema was serious. Out of the 15 bilayer lens scaffolds preserved in anhydrous glycerol at room temperature, 2 remained colourless and transparent, 5 slightly yellowed but still remained transparent, 8 yellowed substantially with a significant reduction in transparency. Out of the 15 bilayer lens scaffolds preserved in anhydrous glycerol at 4℃, 5 remained colourless and transparent, and 10 slightly yellowed while remaining transparent. Of the 15 bilayer lens scaffolds preserved in anhydrous glycerol at -20℃, none of the scaffolds yellowed, therefore, remaining colourless and transparent.

CONCLUSION: FS is a safe and non-toxic bio-gel. It can be used to glue Smile-derived corneal stromal lenses to construct corneal stromal scaffolds with good stability, high transparency and suitable hardness. Anhydrous glycerol at -20℃ is the best preservation condition for corneal stromal lens scaffolds.]]> 2020/3/25 14:45:37 Qing Ye, Ackbarkhan Zacharia, Jia-Yue Ji and Jing Zeng 72true METHODS: TTase KO model was established in this laboratory. TTase KO and WT mice were examined and the lens opacity was classified by using a slit lamp. Each lens was homogenized in lysis buffer and processed for measurement of glutathione(GSH)level. Examination of Protein-GSH mixed disulfides(PSSG)formation in the lens by Western blot analysis. Immunoprecipitation was used to identify the proteins formed PSSG. Dethiolation of lens proteins was carried out using purified recombinant human lens TTase(RHLT).

RESULTS: The slit lamp examination showed an age-dependent nuclear cataract development in both eyes of the WT and TTase KO mice. The onset of cataract was 4mo in the KO mice and 9mo in the WT mice. The GSH loss showed in both groups during aging and was prominent in the TTase KO mice after 9mo old. PSSG in the lenses of both groups showed progressive elevation, whereas the lenses of the KO group had a higher level of PSSG after 9mo. These GSH-conjugated proteins were confirmed as actin and glyceraldehyes 3-phosphate dehydrogenase(GAPDH)by immunoprecipitation and they could be eliminated when the homogenates were treated with RHLT.

CONCLUSION: The results showed that deletion of TTase gene in the mouse could lead to an early age-dependent cataract formation and the PSSG formation in these lenses appeared to link directly to lens opacity. The PSSG could be dethiolated by TTase. This data strengthens that TTase plays an essential role in maintaining lens clarity.]]> 2020/3/13 19:43:56 Jie Zhang, Hong Yan and Marjorie F. Lou 71true METHODS: Thirty-six male rabbits were randomly divided into blank group(A), model group(B), low concentrations butterflybush flower eye drops group(C, 1mg/mL), the medium concentrations drops group(D, 1.5mg/mL), the high concentrations drops group(E, 3mg/mL), and testosterone group(F). In addition to group A, the testes and epididymis were removed from each group to establish a dry eye animal model. After successful modeling, groups A and B remain unchanged. Groups C, D, and E were given different concentrations of butterflybush flower eye drops, 3 times/d. In group F, testosterone propionate was injected into the muscles of the thigh at a dose of 0.5mL/kg once every 3d. Fluorescein staining, Schirmer I test(SⅠt)and tear film break time(BUT)were measured under general anesthesia in each group, eatment. After 4wk of treatment, the rabbits were sacrificed and the conjunctival tissues of the eyes were taken. The expression of IL-1β, mucin 5AC and P38MAPK in the conjunctiva was detected by immunohistochemical staining.

RESULTS: Among low concentrations butterflybush flower eye drops group, the medium concentrations drops group and the high concentrations drops group, the SⅠt value was significantly higher than that of model group, and BUT was significantly longer than model group. The positive staining of corneal fluorescein was significantly improved compared with model group, which was statistically significant(P<0.01). Among IL-1β and P38MAPK in the conjunctiva of high concentrations butterflybush flower eye drops group, the medium concentrations drops group and the low concentrations drops group, the positive expressions were lower than those in model group, and the expression of MUC5AC was higher than that in group model group(P<0.01). In addition, the high concentrations drops group was superior to the low and the medium concentrations drops group.

CONCLUSION: Butterflybush flower eye drops have androgen-like effect. For castrated dry eyes of male rabbits, they can down-regulate the expression of IL-1beta and P38MAPK in dry conjunctival tissue and increase the expression of MUC5AC, thus reducing inflammation infiltration in dry conjunctival tissue and maintaining tear film stability, but their effect is weaker than that of androgen. To the treatment of dry eyes, the middle and high concentration groups of the drops had stronger effects than the low one, and the high concentration group was better than the medium one.]]> 2020/3/13 19:43:56 Xiao-Fang Peng, Qing-Hua Peng, Jun Peng, Ya-Sha Zhou, You-Wei Zhang and Gen-Yan Qin 70true METHODS: Three RNAi sequences were designed to knock down SMP30 target gene RGN expression(KD1-3), and the blank-load sequence was used as the negative control group(NCKD), all of which were used to construct lentiviral vectors to infect SRA01/04 cells. Meanwhile, the uninfected SRA01/04 cells was used as the blank control group(CON). After transfecting SRA01/04 cells, the lentiviral vector with the highest knockdown efficiency was selected by RT-PCR for subsequent experiments. Cells were treated with 15mmol/L CaCl₂ for 24h to simulate a high calcium conditions. BrdU-Elisa assay was used to measure cell proliferation, superoxide dismutase(SOD)assay kit and oxidized glutathione/total glutathione(GSSG/T-GSH)assay kit were used to detect the level of intracellular oxidative stress.

RESULTS: KD1-3 and NCKD lentiviral vectors were successfully constructed to infect SRA01/04 cells with an infection efficiency of about 80%. The knockdown efficiency of KD1-3 group was 93%, 60% and 74%, respectively, KD1 group was selected for follow-up experiment. Under the high calcium conditions, the activity of relative cell proliferation and SOD in KD1 group \〖(2.42±0.08)and(11.69±0.52U/mg)\〗 were significantly lower than that in NCKD group \〖(2.95±0.08)and(31.10±2.24U/mg)\〗 and CON group \〖(2.96±0.25)and(26.33±1.04U/mg)\〗, the ratio of GSSG/T-GSH in KD1 group(70.80±2.34)was significantly higher than that in NCKD group(15.93±3.47)and CON group(20.05±2.45)(P<0.05); there was no significant difference between NCKD group and CON group(P>0.05).

CONCLUSION: Under high calcium conditions, SRA01/04 cells(HLECs)with low expression of SMP30 mediated by shRNA lentivirus resulted in the decrease of the proliferation activity and antioxidant capacity, suggesting that SMP30 may play a protective role in regulating cell proliferation and anti-oxidative stress in HLECs.]]> 2020/1/19 11:26:19 Song-Man Li, Zi-Hao Han, Aint Thu Thu Win, Xi Chen and Hao Liang 69true METHODS: In this experimental study, a total of 72 Kunming mice were randomly divided into control group(Nor), monocular deprivation+ standard environment group(MD+SE), monocular deprivation + enriched environment group(MD+EE)and monocular deprivation+ fluoxetine group(MD+FLX). MD model of mice were established at postpartum 21d, and then fed the mice under SE or EE for 4wk. For the mice in MD+FLX group, they were fed by water with fluoxetine. The visual acuity and flash visual evoked potential of mice in each group were detected. Ultrastructral modifications of synaptic junctions in each group were detected using the electronic microscope. We also applied the molecular biology to study the role of enriched environment in visual cortex of adult amblyopic mice whether through regulating the expression of IGF-1, IGF-1R and IGFBP5.

RESULTS: 1)Visual acuity examination: the successful rate of forepaw-reaching reflex in MD+SE group mice is lower than that in Nor group(P<0.001). Compared to MD+SE group, the successful rate of forepaw-reaching reflex improved in MD+EE group(P<0.001)and MD+FLX group(P<0.001). The difference is not significant between the MD+EE group and MD+FLX group(P=0.816); 2)Flash-visual evoked potential examination: compare to the Nor group, the P2 latency was prolonged(P<0.01), and the P2 amplitude was decreased(P<0.01)of flash-visual evoked potential(F-VEP)in the deprived eye in MD+SE group mice; After raring in enriched environment, the P2 latency was shortened(P=0.003)and P2 amplitude was increased(P=0.000)in the deprivated eye detected with F-VEP, which is not significant in P2 latency and amplitude when compare to MD+FLX group(P>0.05); 3)The structural modifications of synaptic junctions examined by electromicrographs: Compare to the Nor group, the synaptic clefts increased(P<0.01), the synaptic active zone shortened(P<0.01), and the thickness of PSD decreased(P<0.01)in MD+SE group mice. After raring in enriched environment, the synaptic clefts decreased(P=0.0035), the synaptic active zone prolonged(P=0.000)and the thickness of PSD increased(P=0.000)in the visual cortex contralateral to the deprived eye, which is not significant in all of the structural parameters of the synaptic junction in visual cortex when compare to MD+FLX group(P>0.05); 4)IGF-1, IGF-1R and IGFBP5 expression detected by Western-blot: Compare to the Nor group, the IGF-1 and IGF-1R expression in visual cortex contralateral to deprivated eye are both down-regulated in MD+SE group(P<0.01; P<0.01). After raring in enriched environment, the expression of IGF-1 and IGF-1R in MD+EE group was significantly higher than that in MD+SE group(P=0.016; P=0.041), but still lower than that in Nor group(P=0.001; P=0.001). The different expression of IGFBP5 in each group is not significant(P>0.05).

CONCLUSION: Environmental enrichment can improve the visual function through reactivating the plasticity of monocular deprivation amblyopia mice. The mechanism is presumed to be related to the expression of IGF-1 and IGF-1R.]]> 2020/1/19 11:26:19 Yu-Lin Luo, Shi-Shi Luo, Zheng-Hai Liu, Xi-Lang Wang, Li-Juan Tao, Xiao-Ying Wu and Yan-Qiong Tu 68true METHODS: RPE cells were cultured and divided into control group(5.5mmol/L glucose), high glucose group(30mmol/L glucose), low concentration scutellarin group(30mmol/L glucose+1μmol/L scutellarin)and high concentration scutellarin group(30mmol/L glucose+10μmol/L scutellarin), cell scratch experiment observed RPE cells migration, cell survival rate were detected by CCK-8 colorimetry, flow cytometry was used to detect the level of ROS, Hoechst staining was used to observe the proportion of apoptotic cells, and Western blot was used to analyze the changes of Bcl-2 and Bax.

RESULTS: Cell scratch experiment results showed that RPE cells form in low concentration scutellarin group and high concentration scutellarin group were improved than that in high glucose group, cell mobility rate also increased; The CCK-8 results showed that RPE cells survival rate increased to 61.06%±5.59% and 79.81%±7.04% after treated with 1μmol/L and 10μmol/L scutellarin, the difference was statistically significant when compared with high glucose group(40.63%±4.72%, P<0.05); The H₂DCFDA fluorescent probe dying showed that scutellarin reduced ROS generation in RPE cells; Hoechst staining showed that the number of apoptosis RPE cells gradually decreased after treatment with 1μmol/L and 10μmol/L scutellarin; Western blot results showed that scutellarin enhanced the expression of Bcl-2 protein and reduced the expression of Bax protein.

CONCLUSION: Scutellarin could inhibit high glucose-induced oxidative damage and apoptosis in human RPE cells, which provides theoretical support for the research on the therapeutic targets of diabetic retinopathy.]]> 2020/1/19 11:26:19 Hai-Feng Liu, Yong-Hou Zhao and Yuan Gao 67true METHODS: Three kinds of conditioned media(CM)were prepared which including control group(vascular endothelial cells conditioned medium, VCM), normoxic BMSCs-CM group(NCM), and hypoxic BMSCs-CM group(HCM). Then the migration and lumen formation of human umbilical vein endothelial cells(HUVECs)and monkey choroid-retinal vascular endothelial cells(RF/6A)were detected after cultured with the above three kinds of CM respectively for 6-24h.

RESULTS: The numbers of cells migration and the tubes formation(including the total length of the tubes and the numbers of branches)in the hypoxic group(HCM)were increased significantly compared with the control group and the normoxic group(P<0.05). The numbers of migrated RF/6A cells were 19.00±3.61, 32.33±3.06, and 114.00±11.53, respectively in control group(VCM), normoxic group(NCM)and hypoxic group(HCM)after treated for 24h(F=153.3, P<0.001). And the numbers of migrated HUVECs were 76.00±9.54, 122.00±18.68, and 307.70±25.97, respectively in three groups(F=121.5, P<0.001). After incubation of RF/6A cells with three different CM for 6h, the numbers of tubes formation were 12.00±3.00, 37.00±4.58, and 51.00±3.61, respectively(F=81.7, P<0.0001). The results of lumen formation of HUVECs in three groups were similar with that of RF/6A.

CONCLUSION: BMSCs can promote the migration and lumen formation of vascular endothelial cells under hypoxia. This mechanism may play a role in retinal neovascularization.]]> 2019/12/20 14:53:54 Ya-Fen Wang, Jing-Bo Su, Yu-Sheng Wang, Yan-Nian Hui and Chang-Mei Guo 66true in vitro pterygium fibroblasts and its possible mechanism.

METHODS: Pterygium fibroblast obtained from cultured pterygium tissue after surgical excision. Pterygium fibroblasts cells apoptosis level, mitochondrial membrane potential and apoptosis-related factors mRNA and protein expression levels were detected after the induction of berberine with different final concentrations(0, 20, 40, 80μmol/L).

RESULTS: Berberine increased the mitochondrial depolarization level, apoptosis rate, expression level of pro-apoptotic gene Bax, Bad mRNA and protein, and decreased the expression level of Bcl-2 gene mRNA and protein in vitro pterygium fibroblasts cells in a dose-dependent manner.

CONCLUSION: Berberine may induce in vitro cultured pterygium cell apoptosis by increasing mitochondrial depolarization.]]> 2019/12/20 14:53:54 Jing Chen, Wei Li, Hong-Ying Kuang, Rui- Feng Fan and He Sun 65true METHODS: Thirty-six adult male C57BL/6J mice(36 eyes)were randomly divided into three experimental groups and three control groups. Mice in the experimental groups were tail-suspended for 15d(Group one), tail-suspended for 30d(Group two), or tail-suspended followed by returning to normal position for 30d(Group three). Three control groups were similarly fixed with a harness but kept in the normal position for corresponding periods of 15, 30, and 60d. The mice were immediately examined using scotopic ERG(including oscillatory potentials \〖OPs\〗)and fundus fluorescein angiography(FFA)in vivo, and subsequently sacrificed to analyze the retinal histology(methods including immunohistochemistry and TUNEL staining)in vitro. Independent sample t-test was used for data comparison between the same time-point groups.

RESULTS: Following 15-days' tail-suspension, scotopic ERG showed a decline in OPs, but not in the b-wave; the second OP(O₂)showed an amplitude of 197±33μV, which was about 60% of the control level(t=-5.938, P<0.001). Following 30-days' tail-suspension, ERG recovered, with O₂ showing an average value of 264±39μV; when compared to the corresponding control group(308±41μV), no significant difference was observed(t=-1.887, P>0.05). Morphologically, only the 15-days' tail-suspended mice showed FFA with microvascular dilation and tortuosity. Rhodopsin and cone-opsin were almost normal and no apoptotic-positive signals were detected in the retinas of the three tail-suspended groups.

CONCLUSION: Simulating cephalad shifting of bodily fluids as under microgravity, using short-term tail-suspension can affect rodent ERG and retinal microcirculation; however, the change is reversible with no obvious permanent injury observed in the retinas.]]> 2019/12/20 14:53:54 Xu-Feng Dai, Jin-Hua Bao, Xiao-Ping Chen, Wen-Jiong Li, Hai-Xiao Huang and Hao Chen 64true in vitro.

METHODS: Primary rabbit CSCs was cultured in vitro and identified by immunohistochemical staining. using lentivirus(LV)with marker gene enhanced green fluorescent protein(EGFP)transfection rabbit CSCs, the growth status and fluorescence intensity of the transfected cells were observed under an inverted fluorescence microscope. The in vitro animal experiments were randomly divided into 2 groups. experimental group lines of LV-EGFP tag of rabbit CSCs suspension stromal injection, control group amount of normal saline injection corneal stroma, Frozen sections were taken 1wk and 1mo after surgery to observe the fluorescence of transplanted CSCs, and hematoxylin-eosin(HE)was used to observe the tissue morphology of paraffin sections.

RESULTS: LV-EGFP transfected rabbit CSCs showed a small amount of fluorescence after 24h under an inverted fluorescence microscope, with the strongest at 96h and 110h. There was no significant difference in the morphology of the transfected CSCs and normal CSCs. Green fluorescence can be seen in the stromal layer of the cornea in the experimental group at 1wk and 1mo, while there is no green fluorescence in the control group. Paraffin section for 1wk showed obvious epithelial cell hyperplasia and slight corneal edema in the experimental group, and a small amount of inflammatory cell infiltration. 1mo after surgery, the epithelial cell hyperplasia was weakened in the experimental group, and no corneal layer edema was observed. No obvious abnormality was found in the control group for 1wk and 1mo.

CONCLUSION: Extracorporeal corneal stroma transplantation of LV-EGFP labeled rabbit CSCs can survive at least 1mo in the corneal and is compatible with adjacent tissues.]]> 2019/12/20 14:53:54 Lu Zhang, Yan Li, Meng-Yi Li, Xi-Tong Wang, Nan-Yu Li, Zi-Wen Sun and Zhu-Lin Hu 63true METHODS: Human retinal pigment epithelial cells were cultured in three groups, normal control group, high glucose group and intervention group of high glucose aggravated PolyphyllinⅠ, testing after 12h of intervention culture. Normal control group:5.5mmol/L glucose concentration routine culture; high glucose group:25mmol/L high glucose was added to the medium to establish the model; high glucose aggravated PolyphyllinⅠdrug intervention group: high glucose 25mmol/L, 3μg/L PolyphyllinⅠdrug was added to the medium. Immunofluorescence staining assays to observe expression of the Visfatin and VEGF in human retinal pigment epithelial cells; real-time PCR assays for relative expression of Visfatin and VEGF mRNA in epithelial cells; and western-blot assays for Visfatin and VEGF proteins in epithelial cells.

RESULTS: Immunofluorescence detection revealed that Visfatin and VEGF were weakly positive in normal retinal pigment epithelial cells. Visfatin and VEGF were strongly positive in high glucose group. Visfatin and VEGF fluorescence in the drug intervention group was significantly weakened in the higher sugar group. RT-PCR showed that the expression level visfatin mRNA high sugar group was significantly higher than that of normal group and intervention group(t=4.24, 3.89, P<0.05). VEGF mRNA expression was significantly higher in high glucose group than in normal group and intervention group(t=3.53, 2.57, P<0.05). Western-blot results showed that the protein expression of visfatin and VEGF in high sugar group was significantly higher than that in control group and intervention group(t=3.62, P=0.01; t=3.79, P<0.01).

CONCLUSION: The high glucose environment can stimulate the increased expression of Visfatin in retinal pigment epithelial cells, Polyphyllin I can inhibit the expression of Visfatin in retinal pigment epithelial cells in high glucose environment, which may provide a new idea for the treatment of diabetic retinopathy.]]> 2020/11/19 16:34:45 Yi-Nan Shao, Qiang Lu and Xiao-Jing Yang 62true METHODS: Rat primary bone marrow mesenchymal stem cells were extracted and identified; Lewis rats were randomly divided into 3 groups: Control group, Glaucoma group and bone marrow mesenchymal stem cell group(BMMSCs group), and rat glaucoma model were made: the left eye was the experimental eye(glaucoma), the right eye is the control eye, and identified them. Then vitreous cavity transplantation of bone marrow mesenchymal stem cells were performed; HE staining was used to observe the morphology of the retina; The number of retinal ganglion cells was detected by immunofluorescence; TUNEL staining was used to observe the apoptosis of retinal ganglion cells; The expression of IGF1 and BDNF protein in retinal tissue was detected by Western blot.

RESULTS: The intraocular pressure of the experimental eyes of the rats was significantly higher than that of the control eyes(P<0.01), indicating that the glaucoma rat models were successfully constructed; In the Glaucoma group, the retinal nerve fibers were irregularly arranged, the number of retinal ganglion cells and retinal thickness were significantly lower than that of the control group(P<0.01), and the number of apoptosis of retinal ganglion cells was significantly higher than that of the control group(P<0.001); The retinal nerve fiber cells in the BMMSCs group were arranged neatly. The number of retinal ganglion cells and retinal thickness were significantly higher than those in the Glaucoma group(P<0.05), and the number apoptosis of retinal ganglion cells was significantly lower than that of Glaucoma group(P<0.01); The expression of IGF1 and BDNF in the retina of the Glaucoma group was significantly lower than that in the control group(P<0.01), the expression of IGF1 and BDNF in the retina of the BMMSCs group was higher than that in the Glaucoma group(P<0.05).

CONCLUSION:Bone marrow mesenchymal stem cell vitreous cavity transplantation can improve glaucoma in rats and protect retinal ganglion cells.]]> 2020/10/22 16:19:25 Hong-Yi Tian, Bi-Hua Xie and Qing-Li Luo 61true METHODS: Expression of GFAP in hRMECs treated with high sugar(30mmol/mL)and low sugar(5mmol/mL)was detected by qRT-PCR. The high glucose-induced hRMECs cells of silencing GFAP gene was established by lentiviral-mediated method. High glucose-induced hRMECs cells were treated with SRI-011381(TGF-β signaling pathway activator)and dimethyl sulfoxide(DMSO); Expression of GFAP, transforming growth factor-1(TGF-β1), activating transcription factor2(Smad2), Smad3 proteins were measured by Western blot, and cell proliferation and apoptosis were detected by CCK-8 and flow cytometry, respectively.

RESULTS: Expression of GFAP was significantly increased in high glucose treated hRMECs. The high glucose induced hRMECs cell model of GFAP gene silencing was successfully constructed by lentivirus mediation, and the cell proliferation ability was significantly improved, the apoptosis rate is significantly inhibited, and expression of TGF-1, Smad2 and Smad3 proteins in the TGF-β signaling pathway was significantly inhibited after silencing GFAP, while activation of TGF-β signaling pathway could reverse the inhibitory effect of silencing GFAP on the proliferation and apoptosis in high glucose hRMECs.

CONCLUSION: Silencing GFAP gene can promote the proliferation of high glucoseinducedhuman retinal microvascular endothelial cells and inhibit cell apoptosis,the mechanism may be related to the inactivation of TGF-β signaling pathway.]]> 2020/10/22 16:19:25 Liang-Liang Jiao, Wen-Kui Zhu, Cui-Ping Luo and Jian-Ping Lei 60true METHODS:To study the oxidative damage effect of different concentrations of povidone iodine, the cultured epithelial cells were randomly divided into control group, low concentration group, medium concentration group and high concentration group. To study the oxidative damage effect of disinfection time of povidone iodine, the cultured corneal epithelial cells were randomly divided into control group, short time group, medium time group and long time group. Malondialdehyde(MDA)and superoxide dismutase(SOD)were detected by ELISA, cell viability was detected by CCK-8 method and inverted microscope, and apoptotic rate was detected by flow cytometry.

RESULTS: The higher concentration of povidone iodine was associated with the higher MDA content, the lower SOD content, the lower cell activities and the higher apoptotic rate of the corneal epithelial cells, which was in a dose-independent manner. The differences among four groups were statistically significant(all P<0.01). The longer disinfection time of povidone iodine was related with the higher MDA content, the lower SOD content, the lower cell activities and the higher apoptotic rate of corneal epithelial cells, which was in a time-independent manner. The differences among four groups were statistically significant(all P<0.01).

CONCLUSION: The oxidative damage of povidone iodine on corneal epithelial cells were in a dose independent and time dependent manner.]]> 2020/9/17 16:45:29 Yong-Zheng Zheng, Guang-Hui Liu and Ming-Dong Pan 59true METHODS: The human RPE cell line ARPE19 was cultured in low glucose DMEM medium with 10% fetal bovine serum. Experiments were divided into three groups: control group(normal cultured ARPE19), OxLDL group(with 5, 10, 25, 50, 100μg/mL of OxLDL), and LDL group(with 5, 10, 25, 50, 100μg/mL of OxLDL)and cultured 24h. The CCK8 kit(Cell Counting Kit-8)was used to detect cell activity, the flow cytometry was used to detect the percentage of apoptosis and the Western blotting was used to detect the expression of ERS-related proteins and apoptosis-related enzymes. The confocal microscope was used to observe the phagocytosis of Dil-labeled OxLDL(Dil-OxLDL)in RPE cells.

RESULTS: The results of CCK8 showed that when compared with control group, with cell viability of(100±5.637)%, different concentrations(5, 10, 25, 50, 100 μg/mL)of OxLDL could change cell viability significantly(F=41.20, P<0.05), and cell viability of each group was(105.298±9.395)%、(97.106±5.417)%、(77.015±4.055)%、(67.613±3.853)% and(43.872±9.532)%; However, the same concentrations(5, 10, 25, 50, 100 μg/mL)of LDL treatment had no influence on cell viability(P>0.05), and the cell viability changes were(97.55±6.217)%,(99.640±3.586)%,(90.495±2.786)%,(83.552±9.171)% and(90.910±1.429)% respectively. Flow cytometry results showed that OxLDL with concentrations higher than 25μg/mL could induce apoptosis apparently. The apoptosis rates of the blank group, the OxLDL(25μg/mL)group, and the LDL(25μg/mL)group were(5.271±0.519)%,(41.23±1.686)% and(13.07±2.579)% respectively, and the differences among them were statistically significant(F=329.8, P<0.01); The Western blotting results showed that the expression levels of ERS-related proteins and apoptosis-related enzymes in the OxLDL(25μg/mL)group were significantly higher than those in the control group and the LDL group(Caspase-12:F=50.53, P<0.05; GRP78:F=55.60, P<0.05; CHOP:F=38.22, P<0.05; XBP-1:F=53.94, P<0.05; ATF6:F=20.01, P<0.05), while there was no difference between the control group and the LDL group(P>0.05).

CONCLUSION: ERS is involved in the apoptosis of RPE cells induced by OxLDL, and regulating ERS may achieve the purpose of inhibiting RPE cell apoptosis and thus treating RPE apoptosis-related diseases.]]> 2020/9/17 16:45:29 Tong Wu, Kuan-Rong Dang, Jing-Bo Su, Bao-Zhen Lyu, Xin-Ting Lu, Yan-Nian Hui and Hong-Jun Du 58true METHODS: WERI-RB-1 cells with different concentrations of leech extracts(0.02, 0.04, 0.08 and 0.16U/mL)were cultured in vitro for 0, 24, 48 and 72h. The optimal drug intervention concentration and time were selected by CCK-8 method for subsequent experiments. WERI-RB-1 cells cultured in vitro were divided into control group(normal culture medium)and experimental group(culture medium containing leech extract). Flow cytometry was used to detect the effects of drugs on cell cycle and cell apoptosis, and Transwell invasion assay was used to detect the effects of drugs on cell invasion abilit.

RESUITS: According to the detection results of CCK-8 method, the optimal intervention conditions were 0.04 and 0.08U/mL leech extract for 48h. The cells in the leech extract intervention were mainly blocked at G2/M stage, and the positive cell rates in the 0.04 and 0.08U/mL groups were(12.59±5.36)% and(14.79±4.12)%, respectively, which were significantly higher than those in the control group \〖(3.00±2.32)%, P<0.01\〗. The apoptosis rate of the cells in the 0.04 and 0.08U/mL groups was(37.91±3.44)% and(33.05±2.25)%, respectively, which were significantly higher than that in the control group \〖(4.64±2.56)%, P<0.01\〗. The results of Transwell invasion assay showed that the number of cells under the Transwell chamber in the experimental group was significantly than that in the control group, indicating that the leech extract could inhibit cell invasion.

CONCLUSION: The leech extract can inhibit the proliferation and invasion of human retinoblastoma cells and induce cell apoptosis in vitro.]]> 2020/9/17 16:45:29 Yuan-Yuan Li, Yan-Lin Zheng, Xiao-Li Liu, Hui Li, Fang Wang, Miao Zheng, Xin-Yue Zhang and Hong-Jie Ma 57true METHODS: The activity of HLECs was detected by MTT method after incubate with differernt concentration glucose for 24h in vitro to determine the optimal glucose concentration. qRT-PCR and Western blot were used to detect the high glucose pretreatment group and the non-high glucose pretreatment group. The expression of Cyclin D1 in HLECs at different time points after traumatic stimulation was detected.

RESULTS: The viability of HLECs were increased when treatment with low concentration glucose, but the concentration should not exceed 25.5mmol/L, or it will inhibit the activity of HLECs; The reasult of high glucose pretreatment group reveal that the expression of Cyclin D1 is down-regulated in a time-dependent manner within a certain time range. While the expression of Cyclin D1 was irregular in the non-pretreatment group, it was increased at the time point of 12h and 48h. The score treatment can up-regulate the expression of Cyclin D1 in HLECs in a certain degreen.

CONCLUSION: The effects of glucose on HLECs activity and Cyclin D1 experssion are irrugular. Trauma treatment can stimulate the expression of Cyclin D1 in HLECs to some extent.]]> 2020/9/17 16:45:30 Chen Chen, Nan-Nan Zhao, Li-Jun Fu, Zhen-Hua Zhu, Chong Yan, Jia-Chang Zhu and Hao Xiu 56true METHODS: Primary cultured HPFs were identified by Vimentin and CK through immunocytochemical staining. HPFs were divided into control group and DOX group including low, medium and high concentrations(50, 100, 200mg/L). The activity and migration of HPFs were detected by cell counting kit-8(CCK-8)and wound healing assay. The density of VM was observed by three-dimensional cell culture and periodic acid schiff(PAS)staining and compared the differences of VM formation in each group. Western blot was used to analyze the expression of matrix metalloproteinase-9(MMP-9)and vascular endothelial growth factor(VEGF).

RESULTS:Immunocytochemical staining results showed that the cells were spindle shaped, meanwhile, they were positive for Vimentin and negative for CK, which were consistent with the characteristics of fibroblasts. Compared with the control group, the cell activity, mobility, VM density and the expression of MMP-9 and VEGF proteins in the DOX group were significantly decreased(P<0.05). Compared among different concentrations of DOX groups, the differences were statistically significant(P<0.05). Correlation analysis indicated that VM density formed by HPFs was significantly positively correlated with the protein expression of MMP-9 and VEGF(r=0.949, 0.960, all P<0.05).

CONCLUSION: DOX can inhabit HPFs activity, migration, VM density by reducing the expression of MMP-9 and VEGF, suggesting that MMP-9 and VEGF may be the molecular mechanisms of VM formation in pterygium.]]> 2021/8/18 21:32:58 Meng-Xuan He, Jun-Fang Zhang, Ling Yang, Bai Qin, Hong-Wei Gu, Huai-Jin Guan and Hai-Hong Shi 55true METHODS: Fifty-four guinea pigs were randomly divided into normal control(NC), lens-induced myopia(LIM)and LIM+electroacupuncture(LIM+EA). The NC group was fed normally without intervention and the right eye in LIM group and LIM+EA group was coverd with a -6.00D lens to establish a myopia model. At 2 and 4wk, the refraction, axial length and the vessel density of choriocapillaris in groups were measured. The expression and protein content of ET-1, ETAR and ETBR mRNA in groups were detected by the real-time fluorescent quantitative PCR(quantitative polymerase chain reaction, q-PCR), enzyme-linked immunosorbent assay(ELISA)and immunohistochemistry.

RESULTS: At 2 and 4wk, compared with the the NC group, refraction and axial length in LIM group and LIM+EA group had significantly increased(all P<0.001). Compared with the LIM group, the refraction and axial length in LIM+EA group were decreased(all P<0.05). At 2 and 4wk, compared with the NC group,the vessel density of choriocapillaris was decreased(P<0.001)and the ET-1, ETAR and ETBR mRNA and protein levels in choroid were increased(all P<0.05)in LIM group. At 2 and 4wk, compared with the LIM group,the vessel density of choriocapillaris was decreased(P<0.01)and the ET-1, ETAR and ETBR mRNA and protein levels in choroid were increased in LIM+EA group.

CONCLUSION:In LIM guinea pigs, the choroidal blood flow decreased with the increased of refraction and axial length, which may affect ET-1 and its receptors through vascular shear force during the development of myopia. At the same time, electroacupuncture can improve choroidal blood flow through neuromodulation and affects the vascular shear stress to down-regulate the content of ET-1 and its receptor to delay the development of myopia.]]> 2021/7/21 22:23:09 Ting Yu, Hui-Xia Wei, Qing-Mei Tian, Hai-Feng Ji, Ji-Ke Song and Xiao-Feng Xie 54true METHODS: Totally 40 rats were randomly divided into control group(n=10)and DR group(n=30). The rats after successful modeling in DR group were divided into model group(n=10), miR-373 agomir group(n=10), and agomir-NC group(n=10). The right eyes vitreous cavity separately were injected with 200μL normal saline, miR-373 agomir(200nmol)and agomir-NC(200nmol)were treated once a week for 12wk. The expression levels of miR-373 and VEGFA mRNA in each group were detected by RT-qPCR. Dual luciferase experiment was used to verify the targeting relationship between miR-373 and VEGFA. Western-blot was used to detect VEGFA, Bcl-2 related X protein(Bax), b-cell lymphoma-2(Bcl-2), phosphatidylinositol 3-kinase(PI3K)and phospho phosphatidylinositol 3-kinase(p-PI3K), serine threonine protein kinase(AKT), phospho serine protein kinase(p-AKT)protein expression levels.

RESULTS: Compared with the control group, the expression levels of miR-373 and Bcl-2 protein in the retina tissue of model group and agomir-NC group were significantly decreased, and the expression levels of VEGFA mRNA, Bax, p-PI3K and p-AKT protein were significantly increased. Compared with the agomir-NC group, the expression levels of miR-373 and Bcl-2 protein in the retina tissue of miR-373 agomir group were significantly increased(all P<0.05), while the expression levels of VEGFA mRNA and protein, the Bax, p-PI3K, p-AKT protein in the retina tissue of miR-373 agomir group were significantly decreased(all P<0.05). Dual luciferase assay confirmed that VEGFA is the target gene of miR-373.

CONCLUSION: miR-373 can inhibit diabetic retinopathy by targeting VEGFA, which may be related to the inhibition of PI3K/AKT signaling pathway by miR-373.]]> 2021/7/21 22:23:09 Ning Guo, Aiyinu·Nulahou, Qian Bu, Meng Liu and Yan Wang 53true METHODS: Ninety-six healthy C57/6J mice were randomly divided into blank control group, phosphate buffered solution(PBS)group and LC group. The blank control group did not receive any treatment, LC group and PBS group were prepared acute alkali burn models. LC group was given 60mmol/L LC eye drops, and PBS group was given PBS eye drops, 6 times/d, for continuous days from one day before alkali burn. The repair of corneal epithelium was observed by fluorescein sodium staining under slit lamp microscope at 0h, 3 and 7d. On the 3d, the expressions of Ki-67 and IL-1β proteins in cornea were detected by immunofluorescence, the total proteins of corneal epithelial were extracted for Western blot to detect the expression of P63, NLRP3, Caspase-1 and phosphorylation level of STAT3.

RESULTS: The results of corneal fluorescein sodium staining showed that on the 3 and 7d after alkali burn, the percentage of residual corneal epithelial defect area in PBS group compared with LC group was(29.38±6.83)% vs(17.78±4.11)% and(14.23±4.51)% vs(4.10±2.10)%, respectively(P<0.01). The repair of corneal epithelium in LC group was faster than that in PBS group. On the 3d, compared with the blank control group, the expressions of pyroptosis related proteins NLRP3 and Caspase-1 in the corneal epithelium of the alkali burn treated mice were up-regulated, the expression of P63 was decreased, and the p-STAT3/STAT3 level was increased, all the differences were significant except cleaved Caspase-1 of blank control group vs LC group. Compared with PBS group, in LC group, the expression of NLRP3, pro Caspase-1 and cleaved Caspase-1 protein were decreased, P63 was up-regulated, and p-STAT3 /STAT3 was increased, all the differences were significant. Immunofluorescence showed that compared with the blank control group,the expressions of IL-1β and Ki-67 were up-regulated in the alkali burned group. Compared with PBS group, the expression of Ki-67 protein was up-regulated and IL-1β was decreased in LC group.

CONCLUSION: LC can promote the proliferation of stem/progenitor cells in the corneal epithelium of mice and further promote the repair of corneal epithelium after alkali burn by inhibiting the pyroptosis signaling pathway and promoting the activation of STAT3 signaling pathway.]]> 2021/6/24 15:27:59 Bing-Jie Yu, Jun Cheng, Shan Song and Ling-Ling Yang 52true 2021/4/21 21:12:00 Jin Yan, Li Wang, Rong Li, Yang Yang, Wen-Lan Liu, Dan Zhu and Cong Jiao 51true 2021/3/25 20:05:41 Peng-Fei Wang, Chen Shen, Zhe-Hao Yu, Zu-Qing Nie, Zhi-Wei Li, Jie Wen, Meng Li and Xia Cao 50true 2021/3/25 20:05:41 Sha-Sha Yao, Ru-Gang Pan and Chun-Fang Gan 49true METHODS: Fifty SPF rats were randomly divided into five groups: control group with normal feeding, DR group was given the same amount of normal saline by gavage, experimental group A with Pollen Typhae extract 50mg/(kg·d), experimental group B with Pollen Typhae extract 100mg/(kg·d)and experimental group C with Pollen Typhae extract 200mg/(kg·d). Determination of fasting blood glucose in rats of each group. HE staining was used to observe the pathological changes of retina of rats in each group. The expression of IL-6 and TNF-α in the serum of rats were measured by ELISA. The VEGF, VEGFR2 and Ang-1 protein expression in retina tissue were observed by Western blot. The VEGF, VEGFR2 and Ang-1 mRNA expression in retinal tissue were observed by qRT-PCR.

RESULTS: Compared with the control group, the fasting blood glucose, the expression of IL-6 and TNF-α in serum and VEGF, VEGFR2 and Ang-1 protein and mRNA expression in retina tissue in DR group and each experimental group were significantly higher(P<0.05). Compared with DR group, fasting blood glucose decreased in all experimental groups, and the fasting blood glucose of experimental groups B and C was significantly decreased than that of DR group(P<0.05). In addition, the contents of IL-6 and TNF-α in serum, the protein and mRNA expression levels of VEGF, VEGFR2 and Ang-1 in retina tissue in experimental group B and group C were significantly lower than those in group DR(P<0.05). The results of HE showed that the structure of retinal photoreceptor cell layer in DR group was obviously destroyed, the cell edema and gap widened, and the retinal histopathology of rat retina in group B and group C were improved in varying degrees.

CONCLUSION: Pollen Typhae extract can down-regulate the level of inflammation and the expression of VEGF, VEGFR2 and Ang-1 in DR rats, so as to improve the retinopathy.]]> 2021/2/24 14:14:51 Zi-Chao Yang, Yu-Liang Wang and Jing Zuo 48true METHODS: Primary cultured human umbilical vein endothelial cells(HUVEC), 4-7 passages were used for experiments after cell identification. CoCl₂ induced hypoxia to establish the transformation model of endothelial cells into fibroblasts. CCK-8 was performed to detect cell proliferation rate and chose the optimal drug concentration. All cells were divided into 4 groups: control group(FBS-free), CoCl₂(200μmol/L)group, CoCl₂+0.3mg/mL PFD group, CoCl₂+0.6mg/mL PFD group. The protein expression of CD31, VE-cadherin, α-SMA, FSP1, p-p38 and p38 were detected by Western blot. Double immunofluorescence labeling method was used to observe the CD31/α-SMA expression. Wound healing assay detected the cell migration. The q-PCR was applied to detect the mRNA levels of TGF-β1 and SNAI1.

RESULTS: Compared with CoCl₂ group, PFD increased cell proliferation rate and inhibited cell migration significantly under hypoxia(P<0.05). PFD decreased the protein expression of the mesenchymal markers α-SMA and FSP1, and increased the protein level of the endothelial markers CD31 and VE-cadherin(P<0.05). Double immunofluorescence results showed that PFD could reduce the expression of α-SMA and increase the level of CD31(P<0.05). In the process of EndoMT, the p38 protein expression level was stable(P>0.05). PFD down-regulated significantly the high protein expression of p-p38, and high mRNA expression of TGF-β1 and SNAI1 compared with control group(P<0.05). There was no significant difference between the 0.3 and 0.6mg/mL PFD groups in all results above.

CONCLUSION: PFD can inhibit the formation of fibrosis in endothelial cells. TGF-β/p38MAPK signaling pathway might be one of the mechanisms that PFD regulates EndoMT progression. PFD will be expected to become a potential new sight on the treatment of subretinal fibrosis.]]> 2021/1/19 16:56:23 Ding-Ying Liao, Li-Li Fu, Yu-Ping Zheng, Li-Jun Wang, Hong-Song Li, Wen-Yi Zhang, Jian-Ming Wang and Lin Zhao 47true METHODS: Twenty-four adult male SD rats were randomly divided into 6 groups and were put into plateau environment simulation experimental chamber at simulated altitude of 5 000m for 2, 4, 6, 10, 24, 72h respectively. Retinal pathology, HIF-1α and the b wave amplitudes of Max-R of flash ERG were examined by HE,IHC and flash ERG. Twenty-four adult male SD rats were randomly divided into 4 groups and were given respectively placebo, rhodiolarosea, inosine tablet and compound xueshuantong capsule by gavage for 7d. They were put into plateau environment simulation experimental chamber at simulated altitude of 5 000m for 10h. Retinal pathology, HIF-1α and the b wave amplitudes of Max-R of flash ERG were examined by HE, IHC and flash ERG.

RESULTS: In the SD rat model of high-altitude retinopathy, with the increase of experimental time, the ganglion cell layer of rat's retina showed obvious edema and HIF-1α expression increased in the cytoplasm of ganglion cells and core cells. All of them were most obvious at 10h. Compared with the self-comparison of b wave amplitudes of Max-R of flash ERG in each group of SD rats before and after entering in plateau environment simulation experimental chamber, the b wave amplitudes of Max-R in 4h, 6h, 10h and 72h were dramatically decreased(P﹤0.05). And the 2h, 4h(P=0.007), 6h(P=0.008), 10h(P=0.002)were statistically significant differences, the 24h and 4h(P=0.035), the 6h(P=0.040)and 10h(P=0.012)were also statistically significant differences. In the study of protective effect of complex thrombolysis caps on high altitude retinopathy in rat models, the results showed that the rat retinal edema of rhodiolarosea group, inosine tablet group and compound thrombosis capsule group and HIF-1α expression in ganglion cell layer of compound thrombosis group and rhodiolarosea group were significantly reduced comparing with the placebo group. Test for homogeneity of variance and one-way ANOVA were used to test the difference of b wave amplitudes of Max-R of flash ERG in four groups of SD rats after entering in the plateau environment simulation experimental chamber, the results showed the complex thrombolysis caps group(P=0.032)and rhodiola rose group(P=0.001)was significantly lower than placebo group.

CONCLUSION: Compound thrombosis caps may have a protective effect on highaltitude retinopathy in rats by inhibiting the expression of HIF-1α, however, the specific mechanism needs to be further studied.]]> 2021/1/19 16:56:24 Yi Yang, Wen-Fang Zhang, Yu-Ting Li, Xin Zhao, Jin-Hong Zhang, Dong-Mei Zhang and Zhi Li 46true 2O₂-induced oxidative damage of retinal pigment epithelium(RPE)cells.

METHODS:ARPE-19 cells were divided into the control group, H₂O₂ group, different doses of luteolin groups and Nrf2 inhibitor group, and the oxidative damage model of RPE was prepared by 100μmol/L H₂O₂, except for the control group. Cell activity was detected by Methyl thiazolyl tetrazolium(MTT)assay and proper experimental concentration of luteolin was determined. The cell morphology and activity was observed in each group. Cell apoptosis rate and reactive oxygen species(ROS)were detected by flow cytometry, malondialdehyde(MDA)and superoxide dismutase(SOD)were detected by kit method, and the expression of caspase-3, poly adeno-sine diphosphate ribose polymerase(PARP), B cell lymphoma-2(Bcl-2), nuclear factor erythroid 2-related factor 2(Nrf2)and heme oxygenase-1(HO-1)proteins were detected by Western blot.

RESULTS: 100μmol/L luteolin has toxic effects on ARPE-19 cells, so 25μmol/L and 50μmol/L luteolin were selected for subsequent experiments. The cell activity, SOD activity and the protein expression levels of Bcl-2, Nrf2, HO-1 in 25μmol/L and 50μmol/L luteolin groups were significantly higher than the H₂O₂ group(P<0.05). The apoptosis rate, ROS, MDA content and the protein expression levels of Caspase-3 and PARP in 25μmol/L and 50μmol/L luteolin groups were significantly lower than the H₂O₂ group(P<0.05). The cell activity, SOD activity \〖(13.83±1.49)U/mL vs(22.69±1.83)U/mL\〗 and the protein expression levels of Bcl-2, Nrf2 and HO-1 protein expression in the Nrf2 inhibitor group were significantly lower than the 50μmol/L luteolin group(P<0.05). The apoptosis rate, ROS, MDA content \〖(654.96±26.99)vs(446.52±29.42),(3.89±0.29)nmol/mL vs(2.06±0.19)nmol/mL\〗 and the protein expression levels of Caspase-3 and PARP in the Nrf2 inhibitor group were significantly higher than the 50μmol/L luteolin group(P<0.05).

CONCLUSION: Luteolin can improve the oxidative damage of RPE cells induced by H₂O₂, and its mechanism may be related to the activation of the Nrf2/HO-1 pathway.]]> 2020/12/22 18:57:45 Meng Hong, Dao-Xian Hong, Rong-Xian Shi, Cong Wei, Xiao-Li Zhao and Quan-Da Li 45true METHODS: SPF grade SD rats were used to cauterize the scleral surface veins to make a right eye glaucoma model. After successful modeling, different doses(10, 20, 40 mg/kg intraperitoneal injection)of resveratrol were used to intervene. Intraocular pressure was measured 2h after the last administration, and retinal slices were made to observe the survival of retinal ganglion cells(RGC). Retinal plaque to observe the survival of retinal ganglion cells(RGC). Real-time fluorescence quantitative PCR and Western blot were used to detect the mRNA and protein expression of retinal PI3K, Akt, basic fibroblast growth factor(bFGF), and brain-derived neurotrophic factor(BDNF).

RESULTS: The intraocular pressure(30.25±4.25)mmHg in the model group was higher than that in the low-dose group(26.30±4.05)mmHg, the middle dose group(22.31±3.68)mmHg and the high dose group(18.32±3.21)mmHg, and the model group RGC labeling rate(48.25±4.50)% was lower than the low dose group(56.32±5.05)%, middle dose group(66.03±6.68)% and high dose group(78.56±7.82)%(P<0.05). The intraocular pressure in the low, middle and high dose groups decreased in a dose-dependent manner, and the RGC labeling rate increased in a dose-dependent manner(P<0.05). The p-PI3K/PI3K, p-Akt/Akt protein ratio, bFGF, BDNF mRNA and protein relative expression in the model group were lower than those in the low dose group, middle dose group, and high dose group(P<0.05), and the low-dose group, middle dose group and high-dose group increased in a dose-dependent manner.

CONCLUSION: Resveratrol can inhibit the apoptosis of RGC in glaucoma rats and reduce optic nerve damage, which may be related to the up regulation expression of phosphorylated of related proteins in PI3K/Akt signal pathway and the expression of protective gene and protein of optic nerve.]]> 2020/12/22 18:57:45 Lin He, Yun-Juan Jiao, Gao-En Ma, Yan-Hua Li and Xiao-Peng Li 44true METHODS: The inflammation of HCECs was induced by Tumor necrosis factor-α(TNF-α), and the experiment was divided into: control group, TNF-α group and RSV+TNF-α group. The oxidative stress response of HCECs was induced by H₂O₂, and they were divided into normal group, H₂O₂ group and RSV+ H₂O₂ group. MTT assay was used to detect the viability of HCECs; RT-qPCR and enzyme-linked immunosorbent assay(ELISA)methods were used to detect the expression of IL-1, IL-6 and IL-8; Immunofluorescence staining and Western blot were used to observe the nuclear translocation of NF-κB p65. 2',7'-dichlorofluorescein diacetate(DCFH-DA)fluorescent probe was applied to detect the level of reactive oxygen species(ROS).

RESULTS:In the inflammatory response of HCECs, RT-qPCR and ELISA showed that the expression levels of IL-1, IL-6 and IL-8 were increased significantly in the TNF-α group compared with the control group, the above indicators were lower after pretreatment of RSV than those in TNF-α group; Immunofluorescence staining and Western blot showed that the nuclear translocation of NF-κB p65 was increased in TNF-α group, while it was inhibited after pretreatment of RSV. In the oxidative stress response of HCECs, the results of MTT and DCFH-DA fluorescent probe staining showed that H₂O₂ significantly decreased the viability of HCECs and increased the production of ROS in HCECs. After pretreatment of RSV, cell viability increased significantly, and RSV inhibited the generation of ROS in HCECs induced by H₂O₂.

CONCLUSION: RSV has an inhibitory effect on inflammation and oxidative stress damage in human corneal epithelial cells, and it has been confirmed that RSV inhibits inflammation by inhibiting the activation of the NF-κB pathway.]]> 2021/11/22 20:59:15 Lin Zhu, Rui-Fang Han, Pei-Hong Wang and Xuan Li 43true METHODS: Cell counting kit 8(CCK-8)was used to detect the cell viability of human retinal epithelial cells ARPE-19 stimulated with 5, 15, 45, 135mmol/L HG. The ARPE-19 cells were divided into NC group, 45mmol/L HG group, si-NC+45mmol/L HG group, si-lncRNA KCNQ1OT1+45mmol/L HG group, miR-NC+45mmol/L HG group, miR-19a-3p mimics+45mmol/L HG group, si-con+45mmol/L HG group, si-TSHZ3+45mmol/L HG group, pcDNA+si-lncRNA KCNQ1OT1+45mmol/L HG group, pcDNA-TSHZ3+si-lncRNA KCNQ1OT1+45mmol/L HG group. CCK-8 was used to detect cell viability, qRT-PCR was used to detect the expressions of lncRNA KCNQ1OT1, miR-19a-3p and TSHZ3 mRNA, Western Blot was used to detect TSHZ3, activation-cysteine-containing aspartate proteolytic enzyme 3(Cleaved-caspase-3), B-cell lymphoma/leukemia-2(Bcl-2)related X protein(Bax)protein expressions, and enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of oxidative stress indicators reactive oxygen species(ROS)and malondialdehyde(MDA). The dual luciferase activity was used to detect the targeted binding between lncRNA KCNQ1OT1 and miR-19a-3p, miR-19a-3p and TSHZ3.

RESULTS: 15, 45, 135mmol/L HG inhibited the survival rate of ARPE-19 cells, and the subsequent select the HG concentration 45mmol/L with a cell survival rate of about 50%. 45mmol/L HG increased the expression levels of lncRNA KCNQ1OT1, TSHZ3 mRNA, TSHZ3 protein, the apoptosis rate, Cleaved-caspase-3 and Bax protein expressions, ROS and MDA levels in ARPE-19 cells, and reduced cell survival rate and the expression level of miR-19a-3p(P<0.05). Low expression of lncRNA KCNQ1OT1, TSHZ3 or high expression of miR-19a-3p improved the survival rate of ARPE-19 cells induced by HG, and reduced apoptosis rate, Cleaved-caspase-3 and Bax protein expressions, ROS and MDA levels(P<0.05). lncRNA KCNQ1OT1 targeted miR-19a-3p, miR-19a-3p targeted TSHZ3, and lncRNA KCNQ1OT13 regulated the expression of TSHZ3 through miR-19a-3p. The effect of lncRNA KCNQ1OT1 low expression on the survival rate, apoptosis and oxidative stress of ARPE-19 cells induced by HG was reversed by the overexpression of TSHZ3.

CONCLUSION: The low expression of lncRNA KCNQ1OT13 promotes the proliferation of retinal epithelial cells induced by high glucose, and inhibits their apoptosis and oxidative stress through miR-19a-3p/TSHZ3.]]> 2021/11/22 20:59:15 Ni-Hong Zhang, Ding-Shan Hou, Jian-Zhi Qiao, Ke Fu and Hong-Liang Wang 42true 2021/10/22 21:57:36 Man Yang, Wei Tan and Chang Liu 41true 2021/10/22 21:57:36 Fang Chen, Heng Li, Hui You, Yu-Yan Qiu, Ning Wang and Wei Chen 40true 2)on human lens epithelial cells and pyroptosis.

METHODS: Human lens epithelial cells were cultured in vitro and divided into blank control group, H₂O₂ treatment group, and 17β-estradiol+H₂O₂ treatment group. Scanning electron microscope to observe the cytological morphology; immunofluorescence technique to detect Gasdermin D(GSDMD)distribution and fluorescence intensity; CCK-8 to detect cell viability; TUNEL to detect cell pyroptosis; Western-blot to detect Cysteinylaspartate specific proteinase-1(Caspase-1), GSDMD, NOD-like receptor protein 3(NLRP3)protein expression level; ELISA to detect interleukin-1β(IL-1β)expression.

RESULTS: Compared with the control group, the cell viability of the H₂O₂ treatment group was significantly decreased, the expression of Caspase-1, GSDMD, and NLRP3 protein were significantly up-regulated, and the secretion of IL-1β was significantly increased. Compared with the H₂O₂ treatment group, the expression of Caspase-1, GSDMD, and NLRP3 protein in the 17β-estradiol+H₂O₂ treatment group were down-regulated, and the secretion of IL-1β decreased, and it showed a decreasing trend with the increase of estrogen concentration.

CONCLUSION: 17β-estradiol has a protective effect on human lens epithelial cells, and its protective mechanism is related to the inhibition of the pyroptosis process of human lens epithelial cells, and the classical pyroptosis pathway is involved.]]> 2021/9/16 22:17:06 Ying Chen, Gang-Jin Kang, Yan-Xi Wang, Man-Hua Xu and Tao Yang 39true METHODS: Y79 cells were divided into si-NC group(transfected with si-NC), si-circ_0000144 group(transfected with si-circ_0000144), miR-NC group(transfected with miR-NC), miR-502-5p group(transfected with miR-502-5p mimic), pcDNA group(transfected with pcDNA), pcDNA-circ_0000144 group(transfected with pcDNA-circ_0000144), si-circ_0000144+anti-miR-NC group(transfected with si-circ_0000144+anti-miR-NC), si-circ_0000144+anti-miR-502-5p group(transfected with si-circ_0000144+anti-miR-502-5p). Quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect the expression of circ_0000144 and miR-502-5p in retinoblastoma tissues and cells, thiazole blue tetrazolium bromide(MTT)detected cell proliferation, and western blot was employed to determine the expression of nuclear associated antigen Ki67(Ki-67), B cell lymphoma/lewkmia-2(Bcl-2), Bcl-2 associated X protein(Bax), matrix metalloprotease(MMP)-2, MMP-9 protein. Flow cytometry detected cell apoptosis, and Transwell measured cell migration and invasion. Bioinformatics prediction and dual luciferase report experiment analyzed whether circ_0000144 targets miR-502-5p.

RESULTS: The expression of circ_0000144 in 31 cases of retinoblastoma tissue was higher than that of adjacent tissues, and the expression of miR-502-5p was lower than that of adjacent tissues(P<0.05). Compared with the si-NC group, the circ_0000144 expression, OD value, expression of Ki-67, Bcl-2, MMP-2, MMP-9 protein, number of migration and invasion cells of the Y79 cells in the si-circ_0000144 group decreased, and the expression of Bax protein and apoptosis rate increased(P<0.05). circ_0000144 targets and negatively regulates the expression of miR-502-5p. Compared with miR-NC group, miR-502-5p group increased cell apoptosis rate and expression of Bax protein of the Y79 cells, while decreased OD value, number of migration and invasion cells, and the expression of Ki-67, Bcl-2, MMP-2 and MMP-9 protein(P<0.05). Compared with the si-circ_0000144+anti-miR-NC group, cell apoptosis rate and Bax protein expression of the Y79 cells in the si-circ_0000144+anti-miR-502-5p group decreased, but the OD value, number of migration and invasion cells, the protein expression of Ki-67, Bcl-2, MMP-2 and MMP-9 increased.

CONCLUSION: circ_0000144 was highly expressed in retinoblastoma tissue, and inhibiting circ_0000144 can reduce the proliferation, migration and invasion of retinoblastoma Y79 cells, and promote apoptosis through negative regulation of miR-502-5p.]]> 2021/9/16 22:17:07 Ming-Hui Chu, Hai-Yin Chen and Xiao-Li Zhang 38true METHODS:Cultured human retinal pigment epithelial-19(ARPE-19)cells were irradiated with blue light to observe the morphological changes, and the expression of CERKL was detected by PCR and Western blot. ARPE-19 cells were transfected with siRNA-CERKL and pcDNA3.1-CERKL respectively. After exposure to blue light, cell viability was determined by MTT assay, apoptosis was detected by TUNEL assay, content of oxidative stress markers and the expression of SIRT1/E2F1 axis was analyzed. Then siRNA-SIRT1 was transfected into ARPE-19 cells, and the oxidative stress damage of ARPE-19 cells under blue light irradiation was detected again.

RESULTS:ARPE-19 cells gradually contracted into spheres and appeared vacuoles after exposure to blue light. Blue light irradiation led to the increase of CERKL expression level(P<0.05), meanwhile, the rate of cell viability was decreased(P<0.05), the rate of the apoptosis was increased(P<0.05), contents of reactive oxygen species, malondialdehyde and 8-hydroxydeoxyguanosine were increased(P<0.05). Silence of CERKL aggravated this phenomenon, while up-regulation of CERKL could alleviate this change(P<0.05). Up-regulation of CERKL also activated the expression of SIRT1 and promoted the deacetylation of E2F1(P<0.05). Silencing SIRT1 could reverse the alleviating effect of up-regulating CERKL on oxidative stress injury of ARPE-19 cells induced by blue light(P<0.05).

CONCLUSION: CERKL can reduce oxidative stress damage of ARPE-19 cells induced by blue light via activating SIRT1 expression and promoting the deacetylation of E2F1.]]> 2022/7/27 16:29:02 Hai-Rong Zhuang, Zi-Dong Wu, Xue-Hong Chen and Cheng-Jun Li 37true METHODS: Thirty C57BL/6 mice were randomly divided into a negative control group, a myopia model group and a traditional Chinese medicine intervention group, with 10 mice in each group. Except for the negative control group, all mice in the myopia model group and the traditional Chinese medicine intervention group used translucent EP tubes to cover their right eyes to make a form deprivation myopia(FDM)model; The traditional Chinese medicine intervention group gavage Zhujing pill modified suspension 0.546g/(kg·d)(0.15mL/d), the negative control group and the myopia model group were given an equal amount of normal saline(0.15mL/d)for 4wk. At the beginning and the end of the experiment respectively, the right eye diopter of the mouse was measured with a strip retinoscope, measurement of the axial length of the right eye of mouse by A-ultrasound. At the end of the experiment, the right eyes of all mice were taken for detection, and immunofluorescence method was used to locate and detect the activity and migration of the retinal microglia marker(Iba1); Transmission electron microscope observation of autophagosome formation in retinal pigment epithelial cells; Western Blot, real-time fluorescent quantitative PCR(q-PCR)to detect the autophagy marker LC3Ⅱ and p62 protein quantitative and gene expression in retinal tissues.

RESULTS: At the end of the experiment, the refractive power of the right eyes of mice showed that the myopia model group and the traditional Chinese medicine intervention group formed relative myopia, the myopia model group and the traditional Chinese medicine intervention group were significantly lower than those of the negative control group(all P<0.01). At the end of the experiment, the axial length of the myopia model group and the Chinese medicine intervention group were significantly increased compared with the negative control group(P<0.01). Immunofluorescence method for locating and detecting Iba1 showed that the average optical density of Iba1 in the retina of the myopia model group increased the most obviously, followed by the increase in the negative control group, and the decrease in the traditional Chinese medicine intervention group. Compared with the negative control group, the myopia model group increased significantly(P<0.05), and the traditional Chinese medicine intervention group was significantly lower than the myopia model group(P<0.05). It was found that Iba1 migrated to the ganglion cell layer in the myopia model group and the traditional Chinese medicine intervention group. Transmission electron microscopy showed that autophagosomes were observed in the retinal pigment epithelial cells of the myopia model group and the Chinese medicine intervention group. The results of Western Blot and q-PCR showed that the expression of LC3Ⅱ and p62 increased most obviously in the traditional Chinese medicine intervention group, followed by the myopia model group, and the negative control group was the lowest.

CONCLUSION: The results of the study show that modified Zhujing pill may enhance retinal autophagy in mice with FDM by inhibiting the activation of microglia.]]> 2022/6/28 10:46:43 Jie Ma, Ya Mo, Ying-Han Ye, Dan-Ning Long and Xi-Yuan Deng 36true METHODS: A total of 50 healthy C57BL/6 male mice aged 6-8wk were randomly divided into normal control group, iron group, inhibitor group, enhancer group and solvent control group, with 10 mice in each group. The normal control group was injected intraperitoneally with 0.2mL of normal saline, and the other groups were injected intraperitoneally with 50mg/mL iron dextran of 0.2mL, once every 3d. From the 14d, the inhibitor group, the enhancer group and the solvent control group were injected intraperitoneally with the same volume(0.2mL)50mg/kg XMD8-92, 10mg/kg simvastatin and 50% DMSO solvent once a day, respectively. The anterior segment of the eyes was observed under slit lamp microscope on the 7, 14, 28d after intraperitoneal injection, and the ocular surface inflammation index and corneal fluorescein staining score were evaluated. The cornea, conjunctiva and lacrimal gland tissues were taken at 28d for the HE staining and immunofluorescence staining, and RT-PCR were used to detect the expression of macrophage polarization related indexes(CD86, CD206, iNOS, Arg-1); Western blot were used to detect the expression of efferocytosis related signal factors(Gas6, MerTK); ELISA was used to detect the expression of inflammatory factors(IL-1β, TNF-α, MMP-9).

RESULTS: After injection for 28d, compared with the normal control group, the ocular surface inflammatory index and corneal fluorescein staining score were increased in the iron group and the solvent control group. HE staining showed incomplete corneal epithelium, reduced conjunctival goblet cells, unclear lacrimal gland structure and relatively disordered arrangement of cells. In all tissues, the expressions of polarization related indexes of M1 macrophages such as CD86 and iNOS were up-regulated, while those of M2 macrophages such as CD206 and Arg-1 were down-regulated, and the expressions of inflammatory factors such as IL-1β, TNF-α and MMP-9 were up-regulated(all P<0.05). Compared with the iron group and the solvent control group, the ocular surface inflammation index and corneal fluorescein staining score of the inhibitor group were further increased. HE staining showed obvious exfoliation of corneal epithelium, further decrease or even disappearance of conjunctival goblet cells, disorder of lacrimal gland structure and irregular arrangement of cells. In all tissues, the expression of signal factors related to efferocytosis such as Gas6 and MerTK was down-regulated(all P<0.05), the expression of polarization related indexes of M1 macrophages such as CD86 and iNOS and the expression of inflammatory factors such as IL-1β, TNF-α and MMP-9 were further up-regulated(all P<0.05). But the ocular surface inflammation index and corneal fluorescein staining score decreased in the enhancer group. HE staining showed the integrity of corneal epithelial, the increase of conjunctival goblet cells and the improvement of lacrimal gland structure and morphology. In all tissues, the expression of signal factors related to efferocytosis such as Gas6 and MerTK was up-regulated(all P<0.05), and the expression of polarization related indexes of M2 macrophages such as CD206 and Arg-1 was up-regulated, while the expression of inflammatory factors such as IL-1β, TNF-α and MMP-9 was down-regulated(all P<0.05).

CONCLUSION: High-iron environment induces macrophages polarize to M1, which aggravates ocular surface inflammation and tissue damage. Efferocytosis by regulating the polarization of macrophages impact the occurrence of ocular surface inflammation in high-iron environment.]]> 2022/6/28 10:46:43 Jing-Yi Zheng, Xia Meng, Zhi Wang, Jin-Dan Xia, An-Qi Xie, Zhi-Gang Fei and Qi-Guo Xiao 35true METHODS: A total of 40 male C57BL/6 mice(taking the right eye as the experimental eye)were divided into 4 groups by random number table method: There were 10 mice in the control group, each time by intraperitoneal injection of 0.2mL of normal saline; Low-dose group, middle-dose group and high-dose iron group with 10 mice in each group were the model group. Each time, 0.2mL of iron dextran solution with concentrations of 12.5, 25, and 50 mg/mL was injected intraperitoneally. One injection 3d for a total of 28d. We observed the ocular surface inflammation index, corneal fluorescein staining, tear break-up time(BUT)and Schimer I test(SIt)on the 7, 14 and 28d after injection and evaluated the degree of dry eye and ocular surface inflammation. After 28d, the mice were sacrificed for cornea, conjunctiva and lacrimal glands tissue for HE staining, Prussian blue staining and tissue iron detection, to evaluate the inflammatory reaction and iron overload. The expression of inflammatory factors interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α)and matrix metallo proteinase-9(MMP-9)were detected by enzyme-linked immunosorbent assay(ELISA).

RESULTS: Compared with the control group, the mice in the model group showed a series of dry eye symptoms, the inflammation index of ocular surface in mice were increased, the score of corneal fluorescein staining increased, the BUT shortened and the amount of tear secretion decreased(all P<0.05). The cornea, conjunctiva and lacrimal gland tissues of the mice were damaged to varying degrees, the iron deposition on the eye surface of the model group was more serious than that of the control group, and the iron content of the tissue was significantly increased than the control group(all P<0.01). The contents of inflammatory factors(IL-1β, TNF-α, MMP-9)in the cornea, conjunctiva and lacrimal gland tissue of the mice in the model group were significantly higher than those of the control group(all P<0.01). With the increase of injection time and concentration of iron dextran, the degree of dry eye and ocular surface inflammation in mice gradually increased.

CONCLUSION: The mouse iron overload dry eye model was successfully established by intraperitoneal injection of iron dextran, the mechanism may be related to the ocular surface inflammation aggravated by iron overload.]]> 2022/6/28 10:46:43 Xia Meng, Zhi Wang, Jing-Yi Zheng, Jin-Dan Xia, An-Qi Xie, Zhi-Gang Fei and Qi-Guo Xiao 34true 2)-induced hypoxia injury of human retinal pigment epithelial cells(ARPE-19), so as to explore whether astragalus can improve diabetic retinopathy(DR)by anti-oxidative stress.

METHODS: The ARPE-19 hypoxia model induced by CoCl₂ was established and divided into the following 5 groups: normal group(cells were cultured normally without any treatment), hypoxia model group(200μmol/L CoCl₂), blank serum group(200μmol/L CoCl₂+blank serum), low-dose drug-containing serum group(200μmol/L CoCl₂+10% medicated serum)and high-dose drug-containing serum group(200μmol/L CoCl₂+20% medicated serum); CCK-8 detects cell viability; Detect the levels of reduced glutathione(GSH)and malondialdehyde(MDA)in the cell supernatant with a kit; ELISA was used to detect the content of hypoxia-inducible factor-1α(HIF-1α)and vascular endothelial growth factor(VEGF)in cell culture medium; Real-time quantitative PCR(qPCR)to detect the mRNA levels of VEGF, HIF-1α and Prolyl hydroxylase-2(PHD-2); The expressions of VEGF, HIF-1α and PHD-2 were detected by Western Blot.

RESULTS: Hypoxia model of ARPE-19 can successfully establish by CoCl₂ at 200μmol/L. Low-dose and high-dose astragalus-containing serum could inhibit hypoxia-induced ARPE-19 proliferation(P<0.05), increase the GSH level and reduce the MDA content in ARPE-19 with hypoxic injury(P<0.05). Low-dose and high-dose astragalus-containing serum could inhibit the expression of HIF-1α and VEGF in ARPE-19 hypoxic injury supernatant(P<0.05), as well as the mRNA and protein expressions of VEGF, HIF-1α and PHD-2 in ARPE-19(P<0.05).

CONCLUSION: Low-dose and high-dose astragalus-containing serum alleviates the hypoxia injury of ARPE-19 induced by CoCl₂ through anti-oxidant effect.]]> 2022/5/30 15:27:42 Han Hu, Xiao-Qin Wang and Hao Nie 33true METHODS: Twenty-five cases of retinoblastoma tissue specimens with complete clinical data and pathologically diagnosed were collected. At the same time, 9 cases of normal retinal tissue from which the eyeball was removed due to trauma were selected as controls. The qRT-PCR method was used to detect the expression of DLGAP1-AS2 and miR-1193 in normal retinal tissue, retinoblastoma tissue, human normal retinal vascular endothelial cell ACBRI-181, and retinoblastoma cell HXO-Rb44. The si-NC, si-DLGAP1-AS2, miR-NC, miR-1193 mimic, si-DLGAP1-AS2 and miR-1193 inhibitor(co-transfected)were transfected into HXO-Rb44 cells. The dual luciferase reporter experiment was used to detect the targeting relationship between DLGAP1-AS2 and miR-1193. The CCK-8 method, plate clone formation experiment and Transwell experiment were used to detect cell proliferation, colony formation, migration and invasion. Western blot method was used to detect the expression of E-cadherin and N-cadherin protein.

RESULTS: The expression of DLGAP1-AS2 in retinoblastoma tissue was higher than that of normal retinal tissue(P<0.05), while the expression of miR-1193 was lower than that of normal retinal tissue(P<0.05). The expression of DLGAP1-AS2 in HXO-Rb44 cells was higher than that of ACBRI-181 cells(P<0.05), and the expression of miR-1193 was lower than that of ACBRI-181 cells(P<0.05). DLGAP1-AS2 could target the expression of miR-1193. Transfection of si-DLGAP1-AS2 or miR-1193 mimic could inhibit the proliferation, migration and invasion of HXO-Rb44 cells. Co-transfection of si-DLGAP1-AS2 and miR-1193 inhibitor could reduce the effect of transfection of si-DLGAP1-AS2 on the proliferation, migration and invasion of HXO-Rb44 cells.

CONCLUSION: Silencing DLGAP1-AS2 could inhibit the proliferation, migration and invasion of retinoblastoma cells through targeted regulation of miR-1193 expression.]]> 2022/5/30 15:27:43 Hui Li, Ming-Hong Wang, Feng-Ling Liao and Guo-Li Liu 32true METHODS: A hyperglycemic injury model as a research object was established on human retinal vascular endothelial cells. CCK-8 kit was used to detect the effects of ghrelin with different concentrations on cell proliferation under high glucose, so as to screen the optimal concentration of ghrelin. The cells were then divided into normal control group(NC), ghrelin group(ghrelin), high glucose group(HG)and ghrelin+ high glucose group(ghrelin+HG). Cell proliferation was detected by CCK-8 kit, apoptosis was detected by Annexin-APC/7-AAD kit, and the expressions of NLRP3, Caspase-1, IL-1β and IL-18 proteins were detected by western blotting.

RESULTS: Compared with NC group(100.00%±0.00%), the cell proliferation rate of HG group(69.87%±0.68%, P<0.05)was significantly decreased. Compared with HG group, the cell proliferation rate of ghrelin + HG group was significantly increased(92.31%±3.62%, P<0.05). Compared with NC group(4.94%±0.15%), the cellular apoptosis rate of HG group(28.33%±1.37%, P<0.05)was significantly increased. Compared with HG group, the cellular apoptosis rate of ghrelin+HG group(14.24%±0.32%, P<0.05)was significantly decreased. Compared with NC group, the expressions of NLRP3, Caspase-1, IL-1β and IL-18 proteins were all significantly increased(all P<0.05). Compared with HG group, the expressions of NLRP3, Caspase-1, IL-1β and IL-18 proteins in ghrelin + HG group were all significantly decreased(all P<0.05).

CONCLUSION: Ghrelin can protect retinal vascular endothelial cells damaged by high glucose, it may function by inhibiting NLRP3 inflammasome signaling pathway.]]> 2022/5/30 15:27:43 Jian-Bo Li and Yong-Feng Luo 31true in vitro.

METHODS: The hyperglycemia model of HRCECs in vitro was established by simulating diabetic environment with high glucose medium. The cultured cells were divided into normal control group, high glucose control group, high glucose + 20, 40 and 80μmol/L curcumin groups. The proliferation of HRCECs was detected by CCK-8 assay, and the expression of VEGF and NF-κB p65 was detected by Western-blot and immunocytochemistry.

RESULTS: The results of CCK-8 assay showed that high glucose promoted the proliferation of HRCECs significantly compared with the normal control group(P<0.01). Curcumin at different concentrations could inhibit the proliferation of cells significantly in a concentration-dependent and time-dependent manner compared with the high glucose control group after being treated with curcumin at different concentrations for 12, 24 and 48h(P<0.01). The results of Western-blot showed that compared with the normal control group, the expression of VEGF-A and NF-κB p65 in the high glucose control group was increased significantly(P<0.01). Compared with the high glucose control group, the expression of VEGF-A and NF-κB p65 decreased significantly after being treated with curcumin at different concentrations for 12, 24 and 48h, and positively correlated with concentration and time(P<0.01). The results of immunocytochemistry showed that the expression of VEGF in the high glucose control group was significantly higher than that in the normal control group(P<0.01). After 24h of treatment with curcumin,the expression of VEGF was gradually decreased compared with the high glucose control group(P<0.01). There were significant differences in pairwise comparison between each group(P<0.01).

CONCLUSION: Curcumin can inhibit the proliferation and the expression of VEGF and NF-κB p65 of HRCECs induced by high glucose in a concentration-dependent and time-dependent manner, which may be related to its down-regulation of the expression of VEGF and NF-κB p65.]]> 2022/4/24 18:49:22 Biao Xiang, Ying-Zhe Pan, An-Qi Xie, Xi Cheng and Ling Deng 30true in vitro.

METHODS: Human retinal pigment epithelial(ARPE-19)cells chemical hypoxia model was established by cobalt chloride(CoCl₂). CCK-8 method was used to detect the effect of curcumin on the activity of ARPE-19 cells induced by CoCl₂. RT-qPCR and Western blot were used to detect the expression of AKT, HIF-1α, VEGF mRNA and protein in ARPE-19 cells hypoxia model induced by CoCl₂. Cell scratch test, transwell chamber migration test, transwell chamber invasion test and matrigel matrix hose lumen formation test were used to observe the effects of conditioned medium of curcumin in ARPE-19 cells on the proliferation, migration, invasion and lumen formation of human umbilical vein endothelial cells(HUVEC)in non-contact condition.

RESULTS:Chemical hypoxia model of ARPE-19 cells can successfully establish by CoCl₂ at 100μmol/L. CoCl₂ at the final concentration of 100μmol/L can promote the expression of AKT, HIF-1α and VEGF mRNA and p-AKT, HIF-1α and VEGF protein in ARPE-19 cells. Curcumin at the final concentration of 100μmol/L can reduce the expression of AKT, HIF-1α and VEGF mRNA in ARPE-19 hypoxia model. Curcumin at the final concentration of 100μmol/L can reduce the expression of AKT, HIF -1α and VEGF proteins in ARPE-19 hypoxia model. The conditioned medium of low(6.25μmol/L), medium(25μmol/L)and high dose(100μmol/L)curcumin in ARPE-19 cells can significantly inhibit the level migration of HUVEC. The conditioned medium in high dose group can significantly inhibit the vertical migration and cell invasion of HUVEC. The conditioned medium of middle and high dose curcumin in ARPE-19 cells can inhibit the lumen formation of HUVEC.

CONCLUSION:Curcumin at 100μmol/L can protect ARPE-19 cells from hypoxia induced by CoCl₂. Curcumin can inhibit the formation of blood vessels at the cellular level.]]> 2022/3/24 15:57:49 Shui-Ling Chen, Ze-Feng Kang, Wen-Li Chu, Xue-Lian Hao, Fang-Fang Tao, Ming-Ming Zhang and Shu-Jiao Li 29true METHODS:Totally 75 New Zealand white rabbits were randomly divided into three groups, which were implanted with different types of material-Polymethyl methacrylate coated Parylene C(PMMA group), silicone together with injection of Mitomycin C(MMC)(silicon-MMC group)and silicone(silicone group). Aqueous humor were collected at 1, 3d, 1, 2, 3, 4 and 8wk after operation and enzyme-linked immunosorbent assay(ELISA)were utilized to detect the expression of IL-10 in the aqueous humor. The connective tissue surrounding the material were collected at 1, 2, 3, 4 and 8wk postoperatively. Hematoxylin-eosin(HE)staining was applied to evaluate the proliferation of fibroblasts and the infiltration of inflammatory cells. The protein expression and mRNA of IL-10 in the connective tissue were detected by immunohistochemistry and real-time PCR.

RESULTS:Compared with PMMA and silicon-MMC group, silicone group showed significantly increased proliferation of fibroblasts and infiltration of inflammatory cells according to the HE staining result. The result of ELISA showed the expression of IL-10 in the aqueous humor increased significantly at the early stage after surgery, and then decreased gradually,the highest appeared on the third day after operation,and in silicone group there was higher than the other two groups in the early stage postoperatively(1d-3wk)(all P<0.05), and there was no significant difference in the late stages(4-8wk). The protein expression and mRNA of IL-10 in connective tissue were the highest in the first week after operation, decreased gradually at 2-3wk after operation, and increased again at 4-8wk after operation by immunohistochemistry and real-time PCR. And the expression was higher in silicone group than in the other two groups at each time point(all P<0.05). Furthermore, there was a positive correlation between the expression of IL-10 protein and the proliferation of fibroblasts in the late stages(4-8wk).

CONCLUSION: After implantation of glaucoma drainage material, the process of IL-10 increased first, then decreased gradually, and increased again 4wk later, thus IL-10 may be a potential target for inhibiting the scar formation.]]> 2022/3/24 15:57:50 Kang-Yu Zhang, Zheng-Xuan Jiang, Li-Ming Tao, Ning Bao, Kai Li and Yong Liu 28true METHODS: Totally 64 New Zealand white rabbits were randomly divided into 4 groups according to different postoperative medication after PRK operation, including simple PRK group, 14μmol/L DMSO group, 50μmol/L rapamycin group and 100μmol/L rapamycin group. According to the group situation, two hours after the operation, eye drops were given, 3 times a day for 7d. Another 16 rabbits were selected as normal control group. The postoperative inflammatory response and corneal epithelial healing were observed with slit-lamp microscope every day. Haze formation of each group at 1 and 4wk after PRK was collected by slit-lamp microscopy system. Eight rabbits in each group were killed by air embolization 1 and 4wk after PRK, and corneal tissue was extracted and frozen for later use. Immunohistochemistry was used to detect the expression levels of transforming growth factor-β1(TGF-β1), α-smooth muscle actin(α-SMA), and matrix metalloproteinase-2(MMP-2). Real-time PCR was used to detect the relative expression levels of autophage-5(ATG5), autophage-12(ATG12), B lymphocytoma gene-2(Bcl-2)and cysteine aspartic proteinase-3(Caspase3)genes.

RESULTS: Corneal epithelium of all operative rabbits healed completely at 3-4d and no significant difference in healing time between the groups after operation(F=0.745, P=0.530). During the observation period, haze was the most obvious at 4wk after operation in all groups. The haze symptoms were more serious in the simple operation group and the 14μmol/L DMSO group, followed by the 50μmol/L rapamycin group. The haze symptoms in the 100μmol/L rapamycin group were significantly relieved than those in other groups. There was no significant difference in the haze grading with different time points after operation among all groups(all P<0.05). Immunohistochemistry showed that the expression of TGF-β1, MMP-2 and α-SMA was stronger in the operation group and 14μmol/L DMSO group, followed by 50μmol/L rapamycin group, and weakest in 100μmol/L rapamycin group than other groups at 1 and 4wk after operation(all P<0.05). The results of PCR showed that the relative expression of ATG5, ATG12 and Bcl-2 mRNA in 50μmol/L rapamycin group and 100μmol/L rapamycin group were significantly higher than those in simple operation group and 14μmol/L DMSO group at 1 and 4wk after operation(all P<0.05); The relative expression of Caspase3 mRNA in 50μmol/L rapamycin group and 100μmol/L rapamycin group was significantly lower than that in simple operation group and 14μmol/L DMSO group(all P<0.05).