RESULTS: The expression of PCNA in experimental groups was higher than the control groups and there was significant difference(P<0.05). The expression of PCNA gradually decreased as time of alkali burn passed by.

CONCLUSION: Amniotic membrane transplantation can enhanced limbal stem cells proliferation in rats with corneal alkali burn and accelerate the healing of corneal epithelium.]]> Zhi-Rong Liu,Yue Zhang,Guang-Jin Wang and Hui Chen 221true METHODS:Human lens epithelial cells(HLECs)were cultured and sub-cultured in vitro. The cultured HLECs were exposed to H₂O₂ at 300μmol/L over a time course of several hours, with and without pretreatment with β-Estradiol(E₂), ECR, ISR. The HLECs were used for following experiments. The changes of the expression levels of ERK phosphorylation in HLECs which exposed to H₂O₂, with pretreatment with certain concentration of ECR and ISR, were analyzed by flow cytometer(FCM), the signal transduction mechanism of ECR and ISR on antioxidative damage was studied.

RESULTS: The expression levels of ERK phospho- rylation were examined in normal HLECs by FCM,and peaked at 6 hour. The p-ERK levels of H₂O₂ group gradually decreased with a prolongation of treatment time. There were significant differences as compared with control group respectively(P<0.01). The p-ERK levels of E₂ group gradually increased with a prolongation of treatment time, showed significant increase from 6 hour when compared with H₂O₂ group respectively(P<0.01), and peaked at 6 hour. The p-ERK levels of ECR group showed significant increase from 3 hour when compared with H₂O₂ group respectively(P<0.01), and peaked at 12 hour. The p-ERK levels of ISR group showed significant increase from 1 hour when compared with H₂O₂ group respectively(P<0.01), and peaked at 1 hour.

CONCLUSION:The resistance against oxidant-induced injury by E₂, ECR and ISR in HLECs might relate to ERK/MAPK signaling pathway.]]> Ke-Li Zhang,Xiu-Rong Huang,Ming-Xin Qi and Na Guo 220true METHODS: Totally 20 SD rats of 14-day-old were randomly divided into two groups, ten for each group. Rats of experimental group were sutured unilateral eye as monocular deprivation animal models and reared in the same natural light environment with the control group for 45 days. Both two groups received electrophysiological testing, visual cortex ultrastructure was observed by the transmission electron microscope.

RESULTS: Electrophysiologic study on ocular deprived animal model, F-VEP showed typical NPN wave form in normal rats, P1 wave peak latency was short, wave crest was steep and straight. Detection in ocular deprived rats showed that P1 wave peak latency lengthened obviously, wave crest depressed significantly. Observation of ultrastructure of visual cortical neurons in monocular deprived animals by electron microscope showed that it was destroyed seriously.

CONCLUSION: Monocular deprivation can affect rats' visual electrophysiological function. It makes F-VEP P1 wave peak latency of model rats obviously lengthen, amplitude of vibration significantly degrades. Monocular deprivation can destroy rats' visual cortex neurons ultrastructure and this change is the morphological basis of monocular deprivation amblyopia.]]> Xiao-Nan Sun,Jun Tao,Xu-Hong Hao,Li Xu,Ruo-Xi Li and Jin-Song Zhang 219true METHODS: Twenty eyes of New Zealand white rabbits were randomly divided into two groups. After phacoemulsification, the experimental group was implanted with CTR and Crane OV-55C intraocular lens(IOL), and the control group was implanted with Crane OV-55C IOL only. Complications and PCO were observed by slit lamp microscope. Three months after surgery, light microscope and transmission electron microscope examinations were performed.

RESULTS: There was no statistically significant difference between the experimental group and the control group about PCO and Soemmering's ring formation(P>0.05). Pathological evaluations revealed lens epithelial cells(LECs)accumulated at the equatorial capsule forming a large Soemmering's ring and abundant LECs were migrated onto the posterior capsule forming a thick layer in both experimental group and the control group.

CONCLUSION: Sharp-edged CTR fail to retard PCO formation and development in rabbits and it is nessessary to improve it.]]> 2013/8/26 15:29:05 Hai-Hong Shi,Jian Wu and Ling Yang 218true 99Tc-MDP on experimental corneal neovascularization in rats.

METHODS: Chemical cauterization of the cornea was performed by touching alkali. Twenty-four healthy SD rats were divided randomly into two equal groups: experimental group underwent i.p. injection of ⁹⁹Tc-MDP everyday until 14^th day after alkali burn. Control group underwent i.p. injection of normal saline as control. Rats were examined for detection of the first signs of neovascularization. 21 days later, lengths of neovascular and total corneal neovascularization area were assessed.

RESULTS: The time of neovascularization in experimental group was 6.25±1.93 days, the control group was 5.83±1.86 days, and there was no statistically significant difference(P=0.30). The lengths of neovascular were 1.74±0.33mm and 2.82±0.25mm in two groups and the difference was statistically significant. The total neovascularization area was 17.3±3.26mm², 24.8±5.10mm² respectively and the difference between the two groups was statistically significant(P<0.01).

CONCLUSION: ⁹⁹Tc-MDP has a significant effect on inhibition of alkali burn-induced corneal neovascularization.]]> 2013/7/29 14:34:25 Ning Kong,Xiao-He Lu and Bin Li 217true METHODS: SD rats were used for inducing DM models by administrating streptozotocin. Studies were performed with control group, DM group, siNgR group: DM rats intravitreal administration of anti-NgR nucleotide, and siRNA control group: DM rats intravitreally administrated with negative nucleotide. One month later, TUNEL staining was conducted to detect apoptosis of retinal ganglion cell(RGC), level of retinal malondialdehyde(MDA)were detected with kit, and expression of NgR and caspase-3 were observed with Western blot.

RESULTS: The level of retinal MDA in control group, DM group, siNgR group and siRNA control group were 3.74±0.49, 7.21±0.96, 6.41±1.34 and 4.02±0.53nmol/mg prot respectively. A significant increase in the number of TUNEL-positive RGC, the expression of retinal NgR and caspase-3, and the level of retinal MDA were detected in DM and siNgR groups compared with control group(P<0.05). However, the number of TUNEL-positive RGC, the expression of retinal NgR and caspase-3, and the level of retinal MDA showed no difference between control group and siNgR group(P>0.05).

CONCLUSION: NgR induces retinal oxidative stress and up-regulation of retinal caspase-3, playing an important role in the apoptosis of diabetic RGC.]]> 2013/7/1 11:07:54 Zhong-Fu Zuo,Wan-Peng Liu and Xue-Zheng Liu 216true METHODS: Individuals with pathological myopia and control subjects of Chinese Southern Han were selected. Genomic DNA was collected from 5mL peripheral blood, then the exon27, exon36 in the LAMA1 gene were analyzed by polymerase chain reaction(PCR)and direct sequencing. Allele frequencies were tested for Hardy-Weinberg equilibrium(HWE). The χ ² Fisher test was conducted to investigate the genotypic and allelic distri-bution between the pathological myopia and control groups.

RESULTS: In this study, no gene mutation was identified in the exon27, exon36 in the LAMA1 gene.

CONCLUSION: The relationship between pathological myopia and the LAMA1 gene is still waiting for the further proof.]]> 2013/7/1 11:07:54 Jiang You 215true METHODS: Corneas of 32 SD rats were injured with 1mol/L NaOH, then divided into two groups: control and doxycycline-treated. All agents were administered topically 4 times daily. Slit-lamp microscope was performed and inflammatory index was calculated at 3, 7, 14 and 21 days after injury. Then 4 rats were randomly sacrificed and each cornea was divided into two parts, one for histopathology, the other for ICAM-1 ELISA assay.

RESULTS:In control group, the inflammatory index and the number of inflammatory cell was higher than the doxycycline-treated dramatically at all time points(P<0.05). Compared with control group, corneal ICAM-1 expression decreased significantly in doxycycline-treated group at all time points(P < 0.05).

CONCLUSION:Doxycycline may inhibit inflammatory cell infiltration by down-regulating ICAM-1 expression.]]> 2013/6/3 10:33:26 Jing-Bo Zhao,Wen-Jin Zou and Xin-Yu Fu 214true METHODS:The rats(n=87)were randomly divided into three groups. Corneal alkali injury was induced by placing 1mol/L NaOH soaked filter paper on the limbus of right cornea for 20 seconds(group A, n=34)or 40 seconds(group B, n=23), and on the central axis of the right cornea for 40 seconds(group C, n=30)respectively. Corneal transparency, corneal ulceration, and corneal neovascularization were observed and recorded under slit- lamp biomicroscope on day 7 post-operation.

RESULTS: Incidence of corneal ulceration, corneal perforation and positive rate of corneal fluorescein staining in limbal corneal injury groups(group A and B)were significantly higher than that of central corneal injury group(group C)(P<0.05). Incidence of corneal ulceration and corneal perforation in group B was significantly higher than group A(P<0.05). Corneal neovascularization was observed in all three groups.

CONCLUSION: Corneal alkali burns induced by 3mm diameter central cornea injury are fit for the study of corneal neovascularization, while those induced by limbus injury for 20 seconds are fit for the study on limbal stem cells deficiency.]]> 2013/6/3 10:33:26 Hong-Wei Gu and Nan Hu 213true METHODS: MTT was used to assay the different effects on proliferation of HUVEC between the same concentration of Avastin and Lucentis; Transwell was used to measure the different effects on inhibition of HUVEC between the same concentration of Avastin and Lucentis.

RESULTS: MTT showed that A value was significantly different in the respective concentration Avastin groups, Lucentis groups and control group(P<0.05), and there was no significant difference between the same concentration Avastin groups and the Lucentis groups. Transwell analysis showed that the rate of HUVEC migration was significantly different in the respective concentration Avastin groups, Lucentis groups and control group(P<0.05), and there was no significant difference between the same concentration Avastin groups and the Lucentis groups.

CONCLUSION: Avastin and Lucentis could inhibit HUVEC proliferation and migration and inhibition effects of Avastin and Lucentis on HUVEC proliferation and migration enhancement enhangced with the increasing of the drug concentration. There was no significant difference in inhibition effects on proliferation and migration of HUVEC between the same concentration of Avastin and Lucentis in vitro.]]> 2013/6/3 10:33:26 Xin Jin,Bing Chen,Tie-Cheng Liu and Mao-Nian Zhang 212true METHODS: Eighteen 4-week-old healthy domestic cats were included. Twelve of them were used to establish models of monocular deprivation(6 cats)and optical strabismus(6 cats). The other six cats were used as controls. Their binocular visions were detected by P-VEP when they were 6, 10 and 16 weeks old, and the latency and the amplitude of P waves were recorded.

RESULTS: With increasing age, the latency got shortened and the amplitude got enlarged in the control group. The binocular summation was larger than the summation of monocular amplitude. In the monocular deprivation group, the cats had lower amplitude in the right eye than the left eye at 10 weeks old, and the difference was statistically significant(P<0.05); There was no statistically significant difference in the latency. When these cats were 16 weeks old, the latency and the amplitude of the right eye(the amblyopic eye)were obviously lower than the left eye, and partial binocular summation was shown. Compared with the control group, the latency and the amplitude of either eye in the optical strabismus group was not significantly different; however, the binocular summations were remarkably lower than either monocular amplitude when the cats were 10 weeks old and 16 weeks old.

CONCLUSION: P-VEP is an effective approach for evaluation of the binocular vision in animals.]]> 2013/5/6 14:25:10 Lin-Lin Li,Yan Wen and Hua-Qing Gong 211true in vitro and search for a new method to prevent the recurrence after pterygium surgery.

METHODS: HPF was cultivated in vitro. MTT assay was used to detect the effect on proliferation of HPF after incubation for 24 hours, 48 hours and 72 hours in the different concentration of ATRA. The expression of proliferating cell nuclear antigen(PCNA)in each group was detected by immunohistochemical staining.

RESULTS: Administration of 10^-8 mmol/L ATRA for 24 hours could significantly inhibit HPF proliferation in a dose- and-time dependent manner(P<0.05). ATRA could inhibit the expression of PCNA in HPF in a dose-dependent manner(P<0.05).

CONCLUSION:ATRA can significantly inhibit HPF proliferation.]]> 2013/5/6 14:25:10 Xi-Feng Peng,Wen-Hao Jiang,Jian Yan,Jiang-Tao Deng and Fang Cheng 210true METHODS:The retinal pathologic and ultrastructure changes of rds mice treated by CEGI were observed by light and electron microscopy, and the apoptosis rates of photoreceptor cells were determined through TUNEL(tdt- mediated biotin-dUTP nick-end labeling)technique.

RESULTS: Postnatal 14 days(P14)was the best stage of developmental process of rds mice. After 28-56 days, cell number on retinal outer nuclear layer and retinal inner layer reduced, and photoreceptor apoptosis increased gradually. What's more, electron microscopy showed apoptosis nuclear changes and disintegration of inner section and cilia. On 28 days and 56 days, CEGI groups showed thicker retinal cell layers and less apoptotic cells compared with the control group. There was no statistically significant difference in comparison. Nerve growth factor(NGF)had similarity function with CEGI.

CONCLUSION: CEGI can promote growth and development of retinal cells, and have a protective effect on retinal degenerative process of rds mice.]]> Yong-Sheng Yang and Jin-Jin Wang 209true METHODS: The expression of tumor-infiltrating dendritic cells was examined by immunohistochemical staining(S-P method), using CD1a mouse monoclonal antibody in Rb tissues of 31 cases.

RESULTS:The expression of CD1a⁺ cells was found in tumor samples of 23 cases(74.2%)among the total 31 cases. The rates of positive CD1a expression in well-differentiated type was 77.8%(7/9)and in undifferentiated type cancer tissues was 75%(15/20), respectively(P>0.05). The rate of spontaneous regression type was 50%(1/2).

CONCLUSION: There was no correlation between the number of CD1a⁺ cells and tumor grade in Rb. It is difficult to identify whether TIDC was good indicator in assessing tumor prognosis.]]> Song Tang,Xiao-He Lu,Dong-Sheng Zhao and Fang-Wei Ying 208true PAX6 gene has been knocked-down.

METHODS: Four groups PAX6 shRNA lentiviral vectors and the control group lentiviral(pGCL-GFP-shRP1, 2, 3, 4, NC)infect B3 cells. Real Time PCR and Western blot analysis was used to determine PAX6 silence after infection 96 hours. Effective PAX6-silence pGCL-GFP-shRP4 was selected to infect B3 cells, MOI(Multiplicity of infection)= 10 and the changes were observed in the number of the cells. The B3 cells apoptosis was detect by flow cytometry after 48 hours in infection.

RESULTS: RNAi-virus-infected B3 cells decreased in the number of cells with infection prolonged. The flow cytometry results showed that after knock-down the PAX6 for 48 hours, apoptotic peak appeared before the G₁/G₀ phase.

CONCLUSION: Lentivirus-mediated RNA interference can effectively silence PAX6 expression in B3 cells. There is HLE-B3 apoptosis after PAX6 gene knock-down.]]> Yue Huang,Zhi-Qing Li,Zhi-Wei Ma and Hui-Min Sun 207true METHODS: Fresh pig corneas were drilled down by 13mm-diameter trephine. Then the grafts were put into four kinds of cool cryoprotective media by turns and equilibriumed with ‘simplified four-step' method. All grafts were thawed in 40℃. All dexamethasone pretreatment corneas were additionally incubated in the last cool cryoprotective medium which had 0.01mg/mL, 0.03mg/mL or 0.05mg/mL dexamethasone for 18 hours in 4℃ before cryopreservation. Corneas thawed were stained by typan blue and alizarin Bordeaux. Survival rates of endothelia cells were observed.

RESULTS: There were no significant differences in endothelia survival rate among rate-freezing group and every dexamethasone pretreatment group.

CONCLUSION: In 0.01-0.05mg/mL of concentrations, dexamethasone shows no toxic effects. Survival rate of endothelial cells was more than 90%.]]> Fang Liu,Guo-Ping Xiong and Hui-Lian Shen 206true METHODS: RF/6A was randomly assigned into 4 groups(n=6/group): Control group(CONT group, DMEM+10% FBS), high glucose concentration group(HGC group, DMEM+10% FBS+40mmol/L glucose), curcumin group(CUR group, DMEM+10% FBS+30μmol/L curcumin)and curcumin-glucose group(C-G group, DMEM+10% FBS+40mmol/L glucose+30μmol/L curcumin). Five days later, cells were observed with microscope, and cell viability was detected by methyl thiazolyl tetrazolium(MTT). Moreover, agarose gel electrophoresis was introduced to investigate DNA ladder in each group. We also detected expression of tumor necrosis factor-α(TNF-α).

RESULTS: Compared with CONT group, cell viability in HGC group was decreased(P<0.01), showing no obvious difference in CUR or C-G groups(P>0.05). HGC group presented obvious DNA ladder, while no ladder was observed in CONT, CUR or C-G group. Furthermore, compared with CONT group, TNF-α was up-regulated in HGC group(P<0.01), and no difference was detected in CUR group or C-G group(P>0.05).

CONCLUSION: Curcumin attenuates high glucose concentration induced RF/6A apoptosis, maintaining cell viability, which may result from reversing HGC-induced up-regulation of TNF-α.]]> Zhong-Fu Zuo,Qiang Zhang and Xue-Zheng Liu 205true in vitro.

METHODS: Bax expression was detected by ELISA and immunohistochemistry after the lenses of rats were cultivated in MEM culture medium.

RESULTS: The expression of Bax in the mobile microwave groups was significantly higher than the control group(P<0.05)and the expression of Bax accumulated while it was exposed in the mobile microwave.

CONCLUSION: Mobile microwave can induce the expression of Bax in the lens epithelial cells of rat and cause cataract.]]> Kai Shi,Wen-Fang Zhang,Yu Liu and Chun-Li Li 204true METHODS:The riboflavin in rabbit's cornea homogenate was extracted on a C₁₈-SPE cartridge after protein precipitated by heating incubation and adding 10% trichloroacetic acid, and analyzed on HPLC system with an ODS-BP column(5μm, 250mm×4.6mm I.D.)and a mobile phase consisted of 40% methanol and 60% pure water. The flow rate was 0.1mL/min. The spectrophoto- fluorometer was set at wavelength of 425nm for excitation and 525nm for emission.

RESULTS: Riboflavin had a good linearity(r²=0.999 584)existed within range of 1.0×10^-6 to 1.0×10^-4mg/mL. The precision of this method indicated the relative standard deviation(RSD)was 0.86%(n=10), the sample solution stability RSD was 0.51% within 24 hours, and the corneal sample adding standard of the recovery were 99.3%(n=5), RSD=3.7%.

CONCLUSION: This method is specific, accurate and sensitive, and can be used for determination of riboflavin in rabbit's corneal stroma.]]> Teng-Fei Wu and Xiu-Jun Peng 203true METHODS: Alkali injury was induced by application of 1mol/L NaOH to right eyes of New Zealand white rabbits for 30 seconds(n=20). All animals were randomly assigned to A, B groups, each consisting of 10 eyes. Group A received subjunctival injection of avastin after alkali injury immediately in 2.5mg dosage. Group B received subconjuctival injection of normal salt. Biomicroscopic neovascularization was observed on 3, 7, 14, 21 and 28 days. The average length, area and inhibitory rate of corneal neovascularization(CNV)were calculated respectively. Five rabbits were randomly killed in each group on 7 and 28 days and the corneas were taken for histopathological examination.Immunohistochemical studies on the expression of VEGF in corneal paraffin sections were also carried out.

RESULTS:The vessel meshworks of corneal limb were dilated and congested on the 1^st day, then neovascularization(NV)began to invade cornea on 3^rd day, and reached to its developmental peaks between 7 and 14 days, lastly, NV started to regress on 14-21 days after alkali injury. Significant difference(P<0.05)in the area of NV, average length of NV and corneal edema were found respectively between group A and B(P<0.05); the inhibitory rate of CNV ranges from 44.2% to 55% in group A. There were slight epithelium and fiber edema in group A,and disappeared in the late, the fiber arranged straightly and less NV intruded into the central cornea. Immunohistochemical studies indicated that the expression of vascular endothelial growth factor(VEGF)in group A was obvious lower compared to group B on 7 and 28 days.

CONCLUSION:Avastin suppresses corneal neovascu- larization induced by alkali injury on rabbit eyes with subconjunctival injection through decreasing expression of VEGF.]]> Zuo-Hong Wu,Chun-Xia Feng,Jie Tian,Yu-Guang Zhang and Shi-Long Yan 202true METHODS:To produce punctal plugs with polylactic acid. To model the dry eye rabbits by removing of the main lacrimal gland, the third eyelid and Harder's gland; To record the results of Schirmer test and the pyrogen test before and after the implantation of punctal plugs; Schirmer test and pyrogen test were performed before and after punctal plug implantation. The lacrimal plugs histocompatibility and the degradation time were also observed.

RESULTS: It can significantly improve the Schirmer test of dry eye rabbits to implant punctual plugs. No pyrogenic reactions had been discovered. Good histocompatibility was achieved and the plugs would degrade within 2 months.

CONCLUSION:The plugs produced by polylactic acid biodegradable material, which is widely used in biomedical fields, have no toxic effects on rabbits. It's safe and inexpensive, so it's valuable for further research and clinical application.]]> 2013/11/25 11:01:51 Zhi-Yong Wu 201true METHODS: Normal rabbit cornea, axillary lymph node tissue were taken. Get corneal tissue at one week, two weeks after ostrich-rabbit heterogeneous corneal transplantation, fixed by formaldehyde, conventional paraffin embedding sectioning dewaxing after water, gastric enzyme digestion method for antigen repair, use mouse anti rabbit CD4, CD8 monoclonal antibody and fluorescent tagging sheep anti mouse IgG for immunohistochemical assay, and observe the expression of CD4⁺, CD8⁺T lymphocytes in the cornea tissue in fluorescence microscope.

RESULTS: The expression of CD4⁺, CD8⁺T cells in the cornea tissue was negative one week after the ostrich-rabbit heterologous cornea transplantation, CD4 T lymphocytes showed strong positive expression after the second week, CD8 was negative.

CONCLUSION: Indirect immunofluorescence method can be used for determination of related antigen expression in local corneal tissue after heterologous corneal transplantation, and it can lay the foundation for the studies of heterogeneous cornea transplantation rejection mechanism and postoperative drug screening.]]> 2013/10/28 11:29:57 Xian-Ning Liu,Peng-An Wu,Jie Wu,Yan Cheng,Shi-Yin Pan,Xiang-Hua Xiao,Ting-Ting Ai,Na An,Wei Wang and Xiu-Ping Zhu 200true METHODS: Fresh tumor tissues from 22 children with sporadic RB were collected to detect the presence of HPV-DNA using polymerase chain reaction(PCR)and MY09/11 primers.

RESULTS: HPV-DNA was present in 7/22(32%)RB tumors. All the positive cases were sporadic unilateral RB(4 cases were male, 3 cases were female). Two bilateral RB cases were negative for HPV-DNA.

CONCLUSION: The presence of HPV in RB tumors can be a pathogenic factor for sporadic RB. But its role and mechanism in carcinogenesis needs further research.]]> 2013/10/28 11:29:57 Guo-Ying Liu and Hua-Sheng Yang 199true + CD25⁺ regulatory T cell in the peripheral blood of the patients with Graves' ophthalmopathy by detecting the ratio of this cell.

METHODS: This experimental study selected 53 GO patients without exophthalmos, 51 GO patients with exophthalmos, and 51 healthy people were collected. The ratio of CD4⁺CD25⁺ T cell in peripheral blood was determined by flow cytometry.

RESULTS: The ratio of CD4⁺CD25⁺ T cell in GO patients without exophthalmos was lower than that in healthy group(P<0.01). The ratio of CD4⁺CD25⁺ T cell in GO patients with exophthalmos was far lower than that in healthy group(P<0.01). Compared with the group of GO patients without exophthalmos, the ratio of CD4⁺ CD25⁺ T cell of GO patients with exophthalmos was also much lower(P<0.01).

CONCLUSION: The ratio of CD4+ CD25+ T cell in the GO patients decreases and there is autoimmune disorder in patients of this disease. Perhaps this is an important mechanism causing immune suppression damage, which provides a new clue for immunological treatment.]]> 2013/10/28 11:29:57 Meng Xin,Qiang Wang,Lei Zhang,Zhao-Dong Han and Hai-Bo Xue 198true METHODS: Eighteen Wistar rats were used as donors and 36 SD rats were used ad recipients. The recipient was injected by tail with donor IL-10-treated imDC or with untreated imDC. The rats were randomly assigned to three groups, 12 in control group received nopretreatment, 12 in imDC group were injected with imDC(2×10⁶)3d before surgery, and 12 in IL-10-imDC group were injected with IL-10-imDC(2×10⁶)3d before surgery. The survival time of corneal allografts in three groups were observed. The expression of NF-κB was detected by histopathological examination 14d postoperative.

RESULTS: The mean survival time was merely 10.17±2.16d, 19.83±2.25d and 27.57±1.72d respectively in control, imDC and IL-10-imDC groups. There was significantly difference among three groups(P<0.01). Corneas in groups B and C had fewer infiltrating cells, corneal thickness almost normal, and stroma arrangement in order, the center of the grafts showed no inflammatory cellular infiltration. NF-κB cells strongly expressed in control group(P<0.05), and there was no significant difference between groups B and C(P>0.05).

CONCLUSION: IL-10 can restrained DC from mature. Derivatives from donor imDC can markedly prolong the survival time of corneal allografts. NF-κB may play a potential role in the development of corneal graft rejection.]]> 2013/9/23 14:08:04 Jiao Qiu and Jun-Ge Zhou 197true METHODS: Glaucoma rats(18 eyes)were modeled by electric coagulation sclera surface vein and randomized into 3 groups, group A received intravitreal injection of 1mg/0.1mL cannabinoid HU-211 every other day respectively; group B was given intravitreal injection of 0.1mL water every other day, group C was high intraocular pressure(IOP)group, 6 eyes were randomly selected for blank control group(group D). IOP was observed every day. The rats were sacrificed after treatment 4wk, froze retina section, HE stain. The density fluctuation of retinal ganglion cell(RGC)neurons assessment the optic nerve of rat model with chronic high IOP glaucoma were measured.

RESULTS: The apoptosis and damage degree of RGC in group B was obviously higher than that in group A, with statistically significant difference(P<0.05). There was no significant difference in groups B and C(P >0.05).

CONCLUSION: Intravitreal injection of cannabinoid HU-211 shows obvious protective effect on optic nerve in glaucom rat models.]]> 2014/8/19 14:40:38 Hui-Feng Liu,Yuan He,Jun Jia,Ming-Li Ji and Jin-Wei Xi 196true METHODS: Selected 13 patients(16 eyes)with DR and 15 healthy people(15 eyes), the expression of VEGF and its receptors(fms-like tyrosine kinase-1, Flt-1 and kinase insert domain containing receptor, KDR)were evaluated by immunohistochemistry in vitreous. The levels of VEGF, the Flt-1 and KDR in vitreous of patients with DR were examined with enzyme linked immunosorbent assay(ELISA).

RESULTS: Immunohistochemical staining showed that the expression of VEGF, Flt-1 and KDR in vitreous vessel membrane of patients with DR was increased significantly. And the levels of VEGF, Flt-1 and KDR in vitreous of patients with DR were obviously higher than that in the control group(P<0.05 or P<0.01).

CONCLUSION: VEGF, Flt-1 and KDR were widely expressed in vitreous of patients with DR, and were positively related to micro-angiogenesis of DR patients. It proved that VEGF and its receptors played important roles in the occurrence and development of DR.]]> 2014/8/19 14:40:38 Huan-Lian Li,Jin-Wen Zhou and Wei Zuo 195true METHODS: We used immunohistochemistry to evaluate the expressions of PDCD4 and B7-H4 in specimens from 52 optic gliomas in children cases, and analyzed the correlation of PDCD4 and B7-H4 expression with clinicopathological factors children optic gliomas.

RESULTS: PDCD4 expression reduced in optic glioma tissue: PDCD4 expression rate in 52 cases of optic glioma tissues decreasing was correlated with tumor malignancy increase(P<0.01); B7-H4 positive expression increased in optic glioma tissue: B7-H4 expression rate in 52 cases of optic glioma tissues increasing was correlated with tumor malignancy increase(P<0.01); PDCD4 expression was negatively correlated with B7-H4 expression in optic gliomas(P<0.01).

CONCLUSION: Expressions of PDCD4 and B7-H4 may be clinically relevant to tumor malignancy of optic gliomas in children and prevention of metastasis and prediction of prognosis.]]> 2014/7/22 8:27:35 Yan-Tong Liu 194true METHODS: Thirty C57BL/6J mice were randomly divided into two groups. The number of the mice in each group was 15. The mice in experimental group received dark adaptation from 5:00p.m. to 6:00p.m.,and then exposed to LED white light from 6:00p.m. to 7:00p.m. everyday for a month. At 1, 3, 7, 14 and 30d after the beginning, we examed the histology of mice retinas, calculated the thickness of outer nuclear layer(ONL),inner nuclear layer(INL)and analyzed electrophysiology of mice.

RESULTS:One month after experiment, compared to the control group, the latency of Rod-R a wave of the mice in experimental group significantly prolonged, the amplitude of Cone-R b wave of the mice in experimental group significantly decreased and the latency of b wave of the mice in experimental group significantly prolonged(P<0.05).There are no significant difference in the histology of retina, ONL and INL thicknesses.

CONCLUSION: 100lux low intensity white light could give rise to the impairment of the retinal functions in dark-adapted mice.]]> 2014/7/22 8:27:35 Yun-Zhi Lin,Ping Xie and Qing-Huai Liu 193true METHODS: Immunohistochemical S-P staining method was adopted in detecting the expression of VEGF, CD34, Ki-67 and p21 in 62 cases of pterygia and 20 cases of normal conjunctival tissues. Relationship between these markers and clinical pathological characteristics was analyzed.

RESULTS:(1)The positive expression of VEGF, CD34, Ki-67 and p21 in 62 cases of pterygia was 74.2%(46/62), 77.4%(48/62), 66.1%(41/62)and 40.3%(25/62)respectively. The differences were statistically significant compared with normal conjunctival tissues(P<0.05).(2)The expression of VEGF and CD34 in 62 cases of pterygia was correlated with clinical types and stages(P<0.05), and was not associated with sex, age and occupation(P>0.05); the expression of Ki-67 was correlated with clinical stages(P<0.05), and was not associated with other clinical pathological characteristics(P>0.05); the expression of p21 was correlated with clinical stages and pterygium characters(P<0.05), and was not associated with other clinical pathological characteristics(P>0.05).(3)Spearman correlation showed that there was a positive correlation between VEGF and Ki-67(r=0.279, P<0.05), a positive correlation between VEGF and CD34(r=0.299, P<0.05), a negative correlation between VEGF and p21(r=-0.267, P<0.05); it also showed that there was no correlation between any two of CD34, Ki-67 and p21(P>0.05).

CONCLUSION:(1)Overexpression of VEGF, Ki-67, CD34 and low expression of p21 suggest that these markers are concerned with the development and progression of pterygium.(2)Expression of VEGF and CD34 increases along with the increase of clinical types and stages, expression of Ki-67 increases along with the increase of clinical stages, and expression of p21 decreases along with the improvement of clinical types or stages; they suggest that these markers may play important roles in the development and recurrence of pterygium.(3)There is positive correlation between VEGF and Ki-67, VEGF and CD34 as well as negative correlation between VEGF and p21. They suggest that there may be synergistic action between two factors during the development and progression of pterygium.]]> 2014/6/19 9:55:00 Li-Bo Wang,Hai Li,Yu-Zhou Wu,Yi Wang and Kai-Qiang Wu 192true METHODS:The expression difference of TGF-β1, HSP-47 between human pterygium and normal conjunctive tissues were compared by immuno-histochemistry technique.

RESULTS:The positive expression of TGF-β1, HSP-47 was stronger than in normal conjunctive tissues(P<0.05), the TGF-β1 expressed in all layers of pterygium, especially in the squamous epithelium, in the inflammation cells and vascular endothelial cells also expressed. The HSP-47 showed higher expressed in the lamina propria layer of pterygium, and weakly expressed in epithelial layer, no obvious expression in normal conjunctive tissues.

CONCLUSION:Over-expression of TGF-β1 and HSP-47 in pterygium compared to the normal conjunctiva tissues may play a critical role during the occurrence, development and invasion of the pterygium.]]> 2014/6/19 9:55:01 Hui Yan,Ru-Gang Pan,Sha-Sha Yao and Zhi-Rong Yang 191true METHODS: The 30 Wistar rats as donator and 60 SD rats as receptor were used to establish the animal models of penetrating keratoplasty rejection. And 60 SD rats were randomly divided into 3 groups. Donor cornea of Wistar rats preserved in different methods were used separately in 3 groups. The penetrating keratoplasty rejection index(RI), means survival time(MST)of corneal grafts and pathological changes in post-operation were analyzed.

RESULTS: The MST was(10.4±1.70)d in moist-chamber-preserved group(Ⅰ),(12.9±1.81)d in medium-term-preserved group(Ⅱ)and(16.1±2.57)d in cryopreserved group(Ⅲ). The MST in the cryopreserved group was evidently prolonged, showing a significant correlation compared with other two groups(P<0.01). The sections with HE staining revealed that the severity of inflammation in Ⅲ group was reduced compared with that of Ⅰ,Ⅱ group after 10d of keratoplasty.

CONCLUSION: The postoperative rejection of penetrating keratoplasty in rats is decreased and rejection time is delayed in cryopreserved cornea.]]> 2014/5/22 9:13:01 Xiao-Xia Niu,Jing Hong,Yun-Feng Li and Lu-Yang Zhan 190true METHODS: Thirty New-Zealand rabbits were divided into three groups randomly, 10 rabbits for experimental group, 10 for stroma group and 10 for control group. Corneal endothelial cells were removed to establish animal model of corneal endothelial failure. PLEK was performed on the rabbits of experimental group and stroma group, and nothing was transplantated onto the rabbits of control group with the deep layer excised only. Postoperative observation was taken for 3mo. The degree of corneal edema and central corneal thickness were recorded for statistical analysis.

RESULTS: Corneas in experimental group were relieved in edema obviously compared with that in stroma group and the control group, and showed increased transparency 7d after the operation. The average density of endothelial cells was 2 026.4±129.3cells/mm², and average central corneal thickness was 505.2±25.4μm in experimental group, while 1 535.6±114.5μm in stroma group and 1 493.5±70.2μm in control group 3mo after operation.

CONCLUSION:We achieved preliminary success in our study that culturing HUVEC on acellular corneal stroma and performing PLEK for corneal endothelial decompensation. HUVEC transplanted could survive in vivo, and have normal biological function of keeping cornea transparent. This study provides a new idea and a new way clinically for the treatment of corneal endothelial diseases.]]> 2014/5/22 9:13:01 Li Cui,Xiang Ma and Yan-Hui Zhao 189true METHODS: Totally 14 New Zealand rabbits were used in the present study. One side of nasolacrimal duct was obstructed to produce an experimental model and operated the reverse implantation of nasolacrimal duct intubation. Histological changes of the lower segment of nasolacrimal duct mucosa were observed by routine light microscope at 2, 4, 6, 8, 10, 12 and 14wk after the operation.

RESULTS: Compared with the control side, the group of 2 and 4wk after surgery presented the inflammatory cytokine. The group of 12wk after the operation presented isolated granuloma. Group 12 and 14wk presented scattered granuloma. The size of the granulomas was smaller and the density of epithelioid cell and fibroblast were lower in group 12wk than those in group 14wk by HE and Masson trichrome stain.

CONCLUSION: Recurrent Silicone Tube is used to treat nasolacrymal duct obstruction. Nasolacrimal duct can be narrowed and blocked again by granuloma, progressive fibrosis and adhesion of surrounding tissues when tube is in the duct more than 12wk.]]> 2014/5/22 9:13:01 Yun Peng,Lin Ye,Jing-Xian Zhang,Da-Hui Ma and Kun Zeng 188true METHODS: BMSCs were transfected pEGFP/Ang-1, confirming by observation of enhanced green fluorescent protein(EGFP)with fluorescence microscope. Rhesus chorioido-retinal endothelial cells(RF/6A)were cultured with or without pEGFP/Ang-1 transfected BMSCs in transwell under high glucose concentration. Three days after co-cultured, cell vitality and pPKB expression of RF/6A were detected with methyl thiazolyl tetrazolium(MTT)methods and Western blot respectively.

RESULTS: EGFP was expressed in BMSCs successfully transfected with pEGFP/Ang-1. RF/6A co-cultured with pEGFP/Ang-1 transfected BMSCs showed higher viability and pPKB expression compared with RF/6A cultured in condition of high glucose concentration cultured with or without untransfected BMSCs(P<0.01 respectively), showing no difference to control group(P> 0.05 respectively).

CONCLUSION: pEGFP/Ang-1 transfected BMSCs showed protective effects, may via PKB, on RF/6A in condition of high glucose concentration.]]> 2014/4/21 11:07:17 Yang Hou,Fu-Zhi Li,Zhong-Fu Zuo and Xue-Zheng Liu 187true METHODS: RPE cells were cultured with 200μmol/L cobalt chloride(CoCl₂)for 12 hours to imitate chemical hypoxia. Pretreatment of 17-AAG was 1 hour prior to hypoxic insult. Experiment was divided into hypoxic control group, Dimethyl sulfoxide(DMSO)control group and 17-AAG pretreatment group(0.01, 0.10, 0.50, 1.00, 5.00 and 10.00μmol/L). RT-PCR and Western blot analysis were used to examine the expression of ILK in cultured RPE cells.

RESULTS: The density ratios of ILK mRNA and β-actin mRNA of hypoxic control group, DMSO control group and 17-AAG pretreatment groups were 1.32±0.04,1.29±0.03,0.93±0.06,0.70±0.05,0.53±0.03,0.44±0.04,0.32±0.04,0.30±0.03; and the density ratios of ILK protein and β-actin protein were 2.16±0.04, 2.13±0.04, 1.65±0.04, 1.13±0.05, 0.74±0.03, 0.41±0.06, 0.35±0.04, 0.35±0.03. The ILK expression in 17-AAG pretreated groups were inhibited compared to hypoxic control group(P < 0.05), and the decrease was in concentration-dependent manner.

CONCLUSION: Hsp90 inhibitor 17-AAG was effective to inhibit the expression of ILK induced by hypoxia in RPE cells.]]> 2014/4/21 11:07:17 Ye-Qing Wang,Jian Wang,Yong Lü,Li Dong,Hua Mu and Wen-Ping Cao 186true METHODS: Eighty-four Wistar rats were divided into normal control group and at 6h, 12h, 24h, 48h, 3d and 7d group after retinal ischemia/reperfusion. The model of RIRI was constructed by elevating the intraocular pressure. The expression levels of VEGF and MMP-9 was measured using immunohistochemical staining.

RESULTS: No VEGF positive cells were found in normal group and it began to express at 12h, increased gradually and reached maximal levels at 48h after RIRI. MMP-9 positive expression was rarely detected in the normal control group and began to increase at 6h, reached the peak at 24h. There was statistical difference between normal control group and ischemia/reperfusion group(P<0.01)with regard to the expression of VEGF and MMP-9.The expression of VEGF was positively correlated to MMP-9(P<0.05).

CONCLUSION: The high expression of VEGF and MMP-9 is significantly correlated with RIRI and may play an important role in the occurrence and development of retinal ischemic injury.]]> 2014/4/21 11:07:17 Peng-Cheng Wu,Wen-Fang Zhang and Shu Zhang 185true 7 in retinal capillary pericytes rats.

METHODS: The TUNEL test was applied to observe the survival rate of normal retinal capillary pericytes and diabetic retinal capillary pericytes which were affected by P2X₇ agonist BzATP and antagonist OxATP in microscopy.

RESULTS: Activation of P2X₇ receptors caused occurence of apoptosis in retinal capillary pericytes. Compared with normal retinal capillary pericytes, when they were at the same concentration or time, the diabetic retinal capillary pericytes caused much more apoptosis than normal retinal capillary pericytes. The antagonist OxATP could significantly reduce the toxic effects activated by BzATP, and then increased the livability of retinal capillary pericytes.

CONCLUSION:The activity of P2X₇ receptor plays a regulatory role on RPC apoptosis.]]> 2014/3/24 14:01:48 Wei Zuo,Yun-Yun Zhu,Hong-Tao Yu,Huan-Lian Li,Jin-Wen Zhou and Heng Wang 184true METHODS: Six male SD rats and six RCD rats were used, all of which were mature rats. They were stimulated by red, white, blue and green light and the ERG was recorded. The wavelength of red, green and blue light were 625nm, 525nm and 470nm respectively and the white light was mixed by three color light.

RESULTS: The response of ERG in normal SD rat under green and blue light stimulation were stronger than under red and white stimulation. The dark-adapted ERG of RCD rat responsed to color stimulation was similar to the ERG of normal SD rat, but the amplitude under each color stimulation was lower than that of normal SD rat. Light-adapted ERG response was hardly detected waveform.

CONCLUSION: Rat is sensitive to blue and green light, which can be used as a suggested light stimulation in the ERG recording. The ERG of RCD rat is not specific for color stimulation, and at present we could not use color ERG as a diagnostic indicator.]]> 2014/3/24 14:01:48 Qing-Lin Cao,Lei Zhang,Jing An and Zuo-Ming Zhang 183true in vivo study of blood glucose fluctuation injury mechanism, through intraperitoneal injection of glucose to establish blood glucose fluctuation animal models and to simulate blood glucose fluctuation of patients with diabetes.

METHODS: Rats were randomly divided into four groups: normal control group(NC), normal fluctuation group(NF), diabetes mellitus group(DM)and diabetes fluctuation group(DF). Diabetic models were induced through intraperitoneal injection of STZ. A certain amount of glucose was injected in the rats of group NF and DF intraperitoneally three times a day after the model was established, thereby causing blood glucose fluctuations. Full-day blood glucose monitoring was conducted once every two weeks to observe the blood glucose fluctuation situation, according to blood glucose levels drawing the “time-blood glucose concentration” curve. The stability of blood glucose was evaluated from four different aspects, through calculating mean blood glucose(MBG), standard deviation blood glucose(SDBG), largest amplitude of glycemic excursions(LAGE)and Sclichtkrull Mz value.

RESULTS: The full-day blood glucose of rats in the group NC was in a normal range, and it was stable without fluctuation. The blood glucose of rats in group NF was fluctuated between 5-10mmol/L. The blood glucose of rats in group DM were maintained at higher level(>20mmol/L), without great fluctuation extent and DF with sighificant fluctuation. The blood glucose fluctuation in graoup DF was significant. The blood glucose changes of rats in the group NF and DF were significant and regular. The curve of blood glucose fluctuations was relatively stable. All values of rats in group NF, DM and DF were significantly increased compared with group NC. Group DF was increased more significantly, and the comparison with group NF and DM had significant difference(P<0.01).

CONCLUSION: Significant blood glucose fluctuation can be formed through the method of manual intraperitoneal injection of glucose. The method can be used for simulating the blood glucose fluctuation situation of patients with diabetics. This method has the advantages of simple operation, low injury and good repeatability. It can provide experimental basis for in vivo study of blood glucose fluctuation injury mechanism and pathogenesis of diabetic complications.]]> 2014/3/24 14:01:48 Chun-Liu Gai,Jing-Ru Zhao and Xiao-Long Chen 182true METHODS: The 40 corneal acid burned eyes of the 40 SD rats were divided into 4 groups with each group of 10 eyes randomly. Ⅰ, Ⅱ, Ⅲ groups were treated with dexamethasone(1mg/mL)eye drops at three periods after the corneal burns(acute stage, 0-3d; early repair, 3-7d; late repair, 7-21d); the Ⅳ group is treated without dexamethasone. The injured eyes were photographed and dynamically observed with slit lamp microscope, and the histological changes were scrutinized with HE staining.

RESULTS: Eventually, the cornea repairing situation from good to bad is: Ⅲ, Ⅰ, Ⅳ, Ⅱ. After acid burned, corneal epithelial healing and angiogenesis area were different, the comparison between the experimental groups and control group was statistically significant(P<0.01).

CONCLUSION: After acid burned, curative effects are different with using dexamethasone at different periods. Active effects were produced after using dexamethasone in the acute stage and the late stage. However, using dexamethasone in the early stage may hinder the repairing of cornea, or even dissolve the cornea.]]> 2014/2/27 9:12:34 Kai-Wen He,Yan Huang,Ting-Ting Wang,Peng Zhang,Rui-Fu Dong,Ying-Xia Che and Hui-Zhen Ding 181true METHODS: Twenty-six eyes of 13 rabbits were divided into 3 groups randomly. Nine eyes were implanted into vinyl pyrrolidone modification intraocular lenses, 8 eyes were implanted into titanium oxide modification intraocular lenses, 9 eyes were implanted into unmodified intraocular lenses following lens extraction. The intraocular lenses implanted eyes were enucleated in 90d of post operation and cell adhesion on surface of different intraocular lenses were compared under light microscopy.

RESULTS: The cells' size, number and protein films of intraocular lenses on anterior surface of vinyl pyrrolidone modification group were significantly less than that in two control groups.

CONCLUSION: The uveal biocompatibility of hydrophobic acrylic intraocular lenses after single surface modification by monomer vinyl pyrrolidone is significantly improved.]]> 2014/2/27 9:12:34 Na Li,Gui-Qin Wang,Li-Qun Cao,Xiu-Jun Peng,Ping Lu,Qi Huang and Han-Qing Gu 180true METHODS: Sixteen healthy rabbits were randomly divided into 4 groups(groups 1-4), control(vehicle), low(5μg, 2 drops in 5min), moderate(7.5μg, 3 drops in 10min)and high doses(10μg, 4 drops in 15min)of sufentanil-treated groups. Solvent without sufentanil was administered to the left eyes in the vehicle control group(group 1). Meanwhile, 9g/L NaCl solutions were given to the right eyes in all 4 groups at equal drip rate for auto-control. After 7d, ocular toxicity was firstly evaluated.

RESULTS: Observed with naked eye and slit lamp, no changes including corneal opacity, conjunctival congestion, chemosis, eye secretions, iris abnormalities and temporal eye closed was in all groups. With slit-lamp microscope, the counts of corneal endothelium cells in all groups have no statistically significant difference. With light microscope, no pathomorphological injury in the conjunctiva, cornea, corneoscleral junction, iris, ciliary body, retina and optic nerve were found.

CONCLUSION: The ophthalmic application of sufentanil alone(5-10μg)for sedation and analgesia is safe in a short period.]]> 2014/2/27 9:12:35 Hong-Bin Chen,Jiang-Wen Sun and Rong-Xin Chen 179true METHODS: The eighth generation of bovine RVECs was used for the research. The RVECs were cultivated in nutrient solution with different concentrations of puerarin in experimental group and in nutrient solution without drug, growth factor and serum in the control group. Expressions of eNOS and ET-1 in each group were measured by Western blotting method after 48h.

RESULTS:Compared with the negative comparison group, relative expression of eNOS in puerarin group increased(P<0.01), and eNOS expression also enhanced with the increase of the concentration of drugs; the ET-1 relative expressions of puerarin group decreased(P<0.01), and ET-1 expression also decreased with the increase of the concentration of drugs.

CONCLUSION:By up-regulating eNOS expression, down-regulating the ET-1 expression and regulating vascular nitric oxide endothelin(NO/ET)ratio, puerarin can relax retinal vascular smooth muscle.]]> 2014/1/20 9:29:54 Ning-Ling Wu,Zeng-Yuan Zhuang,Qian Sheng,Yi-Ni Wu and Xiao-Qing Guo 178true 2O₃.

METHODS:ACC-2 cells were cultured. The As₂O₃ of different drug concentration gradients(0, 1.0, 2.0, 4.0, 8.0μmol/L)were applied to ACC-2 cells respectively. The changes in △ψm of ACC-2 cells before and after As₂O₃'s inducing(8.0μmol/L for 24h)were detected by flow cytometry with Rh123 staining. Caspase 3 activity was detected by the multifunctional microplate reader.

RESULTS: Rh123 fluorescence intensity in ACC-2 cells was strongest in the control group, while it weakened in ACC-2 cells in 8.0μmol/L As₂O₃ treatment group. The difference between two groups was significant(P<0.05). With the increase of As₂O₃ concentration(0, 1.0, 2.0, 4.0, 8.0μmol/L), Caspase 3 enzyme activity unit in ACC-2 cells gradually increased.

CONCLUSION: As₂O₃ can induce apoptosis of ACC-2 cells by reducing △ψm. Caspase 3 enzyme activity unit of ACC-2 cells gradually increases with As₂O₃ concentration increases, which results in activation of the expression of Caspase 3, and the cells' irreversible apoptosis process coming immediately.]]> 2014/1/20 9:29:54 Yan-Yan Ouyang,Tao Jiang,Meng Gao,Li-Hua Xiao,Yang Zhou,Yue-Li Dun,Gui-Qiu Zhao,Shi-Hai Liu and Ye Liang 177true METHODS: Retinas of 3-month old and 12-month old rats were prepared into semi-thin sections and then observed under transmission electron microscope to evaluate the changes of RPE-photoreceptor complex.

RESULTS: Apoptosis of RPE cells and degenerative changes of RPE-photoreceptor complex were found in retinas of 12-month old rats.

CONCLUSION: RPE-photoreceptor complex degeneration is an early manifestation of the retinal aging.]]> 2014/1/20 9:29:54 Zhen Li,Jie Li,Rong Wang,Lin Li and Yan Lu 176true METHODS: Twenty rabbits(40 eyes)were divided into two groups randomly, the experimental group included 10 cases(10 right eyes), the control group included 10 cases(10 right eyes), and the left eyes were normal group. The corneas of the rabbits were contacted the liquid mustard gas 400μL/L(0.2mL)and 200μL/L(0.2mL)for 4min in the experimental group and the control group respectively. Then the corneal white light and photofluorography at were conducted at 1, 2, 3, 4, 5, 6d; 1, 2, 3wk; 1, 3mo and the corneal center thickness was measured respectively.

RESULTS: The area of corneal epithelium lesions in experimental group degraded from Ⅳ to 0 after 1wk. The lesions were appeared as flake sample repeatedly in 2wk, 3wk, 1mo, delayed to healing. And that area in control group degraded from Ⅲ to 0 after 1wk. The lesions were appeared as pinpoint sample in 2wk, 3wk, 1mo. The central corneal thickness of experimental group was obviously higher than that of the normal and control group in 1-6d. There was statistically significant difference between the two groups(P<0.05). In 1wk-1mo the difference of three group was not statistically significant(P>0.05). In 3mo the experimental and control group were higher than the normal group, but the difference was not statistically significant(P>0.05).

CONCLUSION: Mustard gas of different concentrations can lead different level corneal lesion and corneal thickness change. The corneal epithelium remodeling change appears within 1wk and completely recover in 1mo after the direct corneal contact. Corneal thickness gets right in 3mo.]]> 2013/12/23 10:55:36 Fei Liu,Fei Wang and Fang Liu 175true METHODS: Ten New Zealand white rabbits whose right eyes had ultrasonic emulsification operation were randomly selected. Emulsifying power, time and other parameters were collected. The follow-up time was 3 months. The complications and the formation of PCO were recorded and classified according to Odrich PCO classification system.

RESULTS: One case of 10 rabbits occurred posterior capsular rupture, 1 case died 1 week after surgery. PCO occurred in the remaining 8 rabbits 1 month after operation and range enlarged 2 months later. Dense fibrosis layer and thick fibrosis layer occurred 3 months later.

CONCLUSION: An ideal PCO animal model can be built in rabbit eye. We should pay attention to reduce the complications.]]> 2013/12/23 10:55:36 Yun-Chuan Li,Qian Cao,Lan Li,Xu Zha,Yuan-Ping Zhang,Hong-Mei Dai and Yu-Lin Liang 174true METHODS:Eighty Wistar rats were divided into 3 groups: control group(20 rats), model group(30 rats)and treatment group(30 rats). After streptozotocin(STZ)induced diabetes mellitus in the model group and the treatment group, the treatment group received 2% pyruvate in diet and drinking. The changes of body weight and blood glucose were observed and the changes of glutathione peroxidase(GSH-PX), malonie dialdehyde(MDA), and Na⁺-K⁺-ATPase levels of retinal tissue and retinal ultrastructure were investigated in three groups at 12wk after occurrence of diabetes.

RESULTS:Compared with control group, the body weight of the model group were significantly decreased, the activities of GSH-PX and ATP in the retina of diabetic rats were significantly lower, the MDA was signigicantly higher and significant changes occurred in retinal ultrastructure. Compared with model group, the blood glucose of the treatment group had no significant changes. However, the activities of GSH and ATP in the retina of diabetic rats were higher, the MDA was lower and the retinal ultrastructure was comparatively mild.

CONCLUSION:Pyruvate can alleviate oxidatie stress reaction, improve the energy metabolism of retina, and delay the development of retinopathy.]]> 2014/12/2 9:06:28 Yan-Xiu Qi,Jun-Da Fu,Yu-Qing Wang and Dong-Lan Wang 173true METHODS: HRCECs extracted from human fresher eyeball and cultured in vitro, and ultimately used in the experiment were the growth of 3^rd～4^th cells, the experimental were divided into blank control group, low glucose control group, high glucose control group, high glucose +(50μL/mL, 100μL/mL, 200μL/mL)different concentrations Coix seed oil group. Detecting the multiplication of HRCECs by MTT, the immunocytochemical method was employed to detect the each group HRCECs of VEGF expression.

RESULTS:MTT assay results showed that: different concentrations of coix seed oil acted at HRCECs for 48h, inhibition of cell proliferation was significant difference compared with high glucose control group(P<0.05). Within 48h showed concentration dependence. There was no statistical difference between the low glucose group and high glucose control group(P>0.05). Immunocytochemical assay showed that: 50μL/mL, 100μL/mL, 200μL/mL Coix seed oil acted at HRCECs 48h, the expression of VEGF decreased significantly compared with the high glucose control group(P<0.05), and in a dose-dependent manner. However, in high glucose control group, the expression of VEGF was obvious higher than that of low glucose control group(P<0.05).

CONCLUSION:Coix seed oil can inhibit the HRCECs proliferation and suppress the VEGF expression in high glucose environment.]]> 2014/12/2 9:06:28 Min Li,Yun-Sheng Zhang,Fang Li and Hui-Can Peng 172true METHODS:We amplified the target gene of shuttle vector PHMCMVTSHR289, conjugated the target gene and eukaryotic expression plasmid pcDNA3.1+, and accredited whether pcDNA3.1+/TSHR289 was connected or not by enzymatic digestion and sequencing. Cationic liposomes encapsulated the recombination plasmid pcDNA3.1+/TSHR289.

RESULTS: Recombination plasmid pcDNA3.1+/TSHR289 digested with enzyme HindIII and the fragment through 0.8% gel electrophoresis showed 512bp strip. Recombination plasmid pcDNA3.1+/TSHR289 were found synonymous mutation through forward(AAC to AAT)and reverse sequencing(GCG to GCT). The volume ratio of cationic liposomes and recombinant plasmid was 3:1.

CONCLUSION: It is successful to construct the recombination plasmid pcDNA3.1+/TSHR289 by accredit it through enzymatic digestion and sequencing.]]> 2014/12/2 9:06:28 Yu-Li Yang,Qing-Li Luo and Hong-Bin Lü 171true METHODS: DR rats were fed with Shuangdanmingmu capsule. Then the blood glucose and glycated hemoglobin were determined. Immunohitochemical staining was used to observe the expression of structure of retino and pancreas.

RESULTS: Compared with the control group, the blood glucose and glycates hemoglobin in Shuangdanmingmu group were decreased after 2mo of treatment(P<0.01). The layers tissue of retina was less edema and gradually arranged in Shuangdanmingmu group. The structure of pancreas was clear basically. Fat cells, infiltrati on of lymphocytes and fibrosis hyperplasia appeared in the interacinar and interlobular connective tissue. Some acinar and islet cells of fatty rat pancreatic tissue had fatty or vesicular degeneration.

CONCLUSION: Shuangdanmingmu capsule can decrease blood glucose and glycates hemoglobin in DR rats effectively and improve the ultrastructure of retina and pancreas tissue.]]> 2014/10/31 11:03:29 Yu-Hui Qin,Wen-Juan Li,Xi Zhang,Zong-Shun Dai,Xiao-Liu Chen,Ya-Sha Zhou,Yan-Jun Ling and Bing Zheng 170true METHODS: Thirty guinea pigs(3-wk old)were received -10.00D rigid contact lenses(RGP)on the right eye and without wearing glasses on the left eye. Diopter change with cycloplegic retinoscopy, ultrastructural features of the retina with transmission electron microscopy were observed, the apoptosis of retinal cell by TUNEL immunofluorescence staining after 3wk were detected.

RESULTS: Eye diopter difference was 3.28±0.21D in the right eye and 1.55±0.23D in the left eye before and after induction. There was significantly statistical difference(P<0.01); Retinal ultrastructural features were showed with transmission electron microscopy(TEM)the retinal disk membranes edema, some shedding, mitochondrial swelling and deformation, small amount of vacuolar changes in the inner and outer nuclear layer cells. Some characteristics of apoptosis were also be seen such as cell membrane irregular contractions, irregular nuclear chromatin aggregation. The incidence of retinal cell apoptosis was(2.42±1.24)% in the right eye and(0.29±0.08)% in the left eye after induction. There was significantly statistical difference(P<0.01).

CONCLUSION: Accompanied by abnormal apoptosis of retinal cells during the formation of defocus-induced myopia in guinea pigs.]]> 2014/10/31 11:03:29 Hong-Wei Zhao,Jing-Nan Liang,Ling Luo,Chuang Nie,Feng-Hua Bai and Yi Liu 169true 2 in retina model. At the same time to clarify the function of TGF-β₂ in form deprivation myopia model.

METHODS: A facemask was worn on the right eyes of guinea pigs to develop form deprivation myopia. Before and after experiment, refraction was measured using retinoscope, and ocular axial was determined by A-scan ultrasonography. Morphologic alternations of the retina were observed by light microsope. The expression of TGF-β₂ in the retina was determined by immunohistochemistry.

RESULTS:Retina of myopic eyes became thinner, and inner nuclear layer of retina had few layers, nuclear of outer nuclear layer became smaller and circular, at the same time, they arranged in disorder. The immunohistochemisty results showed that expression of TGF-β₂ in model eyes was lower than that in control eyes(P<0.05).

CONCLUSION: Covering eyes with facemasks is effective, simple and convenient in making myopic model. The retina of the model eye appeares degeneration. TGF-β₂ plays an important role in the course of development in form deprivation myopia.]]> 2014/10/31 11:03:30 Ling-Xiao Zhou,Li-Lun Wang and Lin Zhang 168true METHODS: Human Tenon capsule fibroblasts were cultured in vitro after glaucoma filtration surgery. The third to sixth passage of cell were treated by 600nmol/L TSA or none. Cell viability measured by MTT assay after 1, 2 and 3d respectively. The expressions of HDAC1 and HDAC2 were analyzed by Western blot 2d after TSA treatment.

RESULTS: Compared to the control, cell viability decreased significantly after treatment with TSA at 1d(P<0.05), presented time-dependent manner. The expression of HDAC1 and HDAC2 significantly reduced in TSA-treated HTF compared with control cells at 2d after TSA treatment.

CONCLUSION: TSA inhibits the proliferation of Tenon capsule fibroblast by inhibiting the expression of HDAC1 and HDAC2, and reduces subconjunctival scar formation.]]> 2014/10/31 11:03:30 Xiao-Yan Li,Ying Deng and Jian-Gang Yang 167true METHODS:Thirty-five patients with pterygium who underwent simple excision and 10 cases of normal conjunctival tissues were enrolled in this study. Immunofluorescence was performed in pterygia tissue and normal conjunctiva, detecting the expression of HIF-1α and Pax6.

RESULTS: The positive expression rate of HIF-1α was 66%(23/35)and 10%(1/10)in pterygia tissue and normal conjunctiva, respectively. The difference of expression between pterygia tissue and normal conjunctiva was statistically significant(P<0.05). Expression of Pax6 was full-thickness nuclear in normal conjunctiva; however, expression of Pax6 decreased and even was negative in pterygium. The difference of expression in two groups was statistically significant(P<0.05).

CONCLUSION: HIF-1α is highly expressed in pterygium, suggesting that it may be involved in the occurrence and development of pterygium. Pax6 gene downregulated in pterygium epithelial cells, suggesting that epithelial cells are squamous metaplasia.]]> 2014/10/31 11:03:30 Xiu-Xia Yin and Qian-Yan Kang 166true 2O₂ and the role of miRNA in oxidative stress induced apoptosis.

METHODS: HLE cell line HLE-B3 was treated by 100μmol/L H₂O₂ for 24h and the total RNA were isolated by Trizol reagent. miRNA profile was generated by miRCURY^TM LNA microRNA Array. The target genes of differentially expressed miRNAs were predicted by bioinformatics software.

RESULTS:Twenty-eight miRNAs showed significantly differential expression after H₂O₂ treatment, 18 miRNAs upregulated and 10 miRNAs downregulated. The differentially expressed miRNAs may involve in apoptosis of lens epithetium and development of cataract through targeting BCL2L2 and MIP.

CONCLUSION: H₂O₂ can induce dramatically changes in miRNA profile of HLE, which may play a pivotal role in the pathogenesis and development of cataract.]]> 2014/9/22 14:18:22 Ling Bai,Peng Li,Ling Chen,Ling He and Feng Wang 165true METHODS: Male SD rats were divided into four groups randomly, control group, diabetic group, apocynin group and puerarin group. The diabetic group were replicated by single injection of STZ(65mg/kg, ip). The expression of p22, p47, p67, Bax/Bcl2, Caspase 3 and P53 proteins were detected by Western Blotting.

RESULTS: The diabetic rats were replicated successfully and the expression of Bcl2 was downregulated while the expression of p22, p47, p67, Bax, Caspase 3 and P53 were upregulated in diabetic group with a significant statistical differences when compared with control group(P<0.05). Apocynin and prerarin can reverse the abnormal expression of the aforementioned proteins dramatically(P<0.05).

CONCLUSION: NADPH oxidase mediated oxidative stress and P53, Bax/Bcl2 mediated apoptosis are involved in the pathogenesis of diabetic cataract and puerarin can alleviate cataract greatly by inhibiting the aforementioned signal pathway.]]> 2014/9/22 14:18:22 Li Wan,Wen-Bin Liu,Ye-Yu Shen,Qiu-Li Yu and Jing-Jing Zhang 164true METHODS: Subconjunctival injection TSA, mitomycin C(MMC)and PBS during the filtering surgery in rabbits. The morphologic changes of filtration blebs were evaluated by Krofeld score method postoperatively days 3, 7, 14, 21, and 28.

RESULTS: TSA induced filteation bleds were elevated diffusely within 14d and cystic blebs formed 28d, filtration bleb score was significantly higher in TSA group than that in PBS group.

CONCLUSION: TSA can keep the aqueous humor outflow by inhibiting scar formation and prolong the existence of the filtration bleb.]]> 2014/9/22 14:18:22 Xiao-Yan Li,Ying Deng and Jian-Gang Yang 163true METHODS: Experimental model of high intraocular pressure was induced unilaterally in 44 C57BL/6 mice. The fellow eye served as a control. TONO-PEN AVIA Tonomter was used to measure intraocular pressure(IOP)to guarantee IOP value at 1, 2, 4, 8wk. Slit-lamp biomicroscopy was performed throughout the period and the structural changes were assessed histologically. And then, their eyes were enucleated, postfixed, cryoprotected, and embedded in optimal cutting temperature medium. After hematoxylin and eosin stain(HE stain), cryosections of the retina were observed under light microscope. TdT-mediated biotin-dUTP nick end labeling(TUNEL)was performed on the retinal sections to determine apoptosis rate.

RESULTS: IOP of laser-treated eyes was significantly higher than that of control eyes from 1-8wk(P<0.05). The highest IOP was 31mmHg, but only one eye. The IOP was mainly around 20mmHg. In laser-treated eyes, the angle of anterior chamber were narrow. Number of cells in the inner nuclear layer and retial gangllion cell layer was slightly lower than that in control eyes at 2wk, but by 4 and 8wk the number of cells was significantly lower than that in the control contralateral eyes.

CONCLUSION: The laser photocoagulation of limbus causes chronic elevation of IOP and this method may be a promising experimental model for the investigation of biological mechanisms of glaucomatous retinal ganglion cell damage.]]> 2014/9/22 14:18:22 Yue He,Shu-Guang Zhang,Yuan-Sheng Yuan,Yan Li,Hong-Bin Lü,Jin-Hua Gan and Li Mao 162true METHODS: Human RPE cell line D407 was treated by 100, 200, 400μmol/L H₂O₂ for 24h and harvested to isolate total RNA by Trizol reagent. The expression difference of D407 cell miRNA after processing of different concentrations was generated by Exiqon miRCURY LNA^TM microRNA expression profile chip and the changes after processing of different concentrations were conducted by Hierarchical Clustering analysis. The results of chips were verified through Stem loop realtime PCR, and the target genes of identified miRNAs were predicted by bioinformatics software.

RESULTS: Among the 1 425 known miRNAs listed on microarray, 367 miRNAs showed differential expression after H₂O₂ treatment. The Treeview Clustering showed that 17 miRNAs, including miR-31, were downregulated along with the increase of H₂O₂ concentration. Meanwhile, 7 miRNAs, including miR206, were upregulated. The results of qRT-PCR further validated the better results of microarray.

CONCLUSION:The miRNA expression of human RPE is dramatically changed after H₂O₂ treatment. miRNA adjusts the molecules level of micrornas transcription and it is involved in cell oxidative stress reaction, and miRNA may play a pivotal role in the pathogenesis and development of AMD.]]> 2015/8/27 11:19:42 Hou-Cheng Liang,Ling Chen,Hai-Xiao Feng,Ling He,Liang Yao and Ling Bai 161true METHODS: Twenty healthy adult New Zealand rabbits were used in the study. The rabbits were randomly divided into two groups, group A received single administration of 1.25mg bevacizumab(1.25mg/0.05mL)by intravitreal injection, group B received single administration of 5mg bevacizumab(5mg/0.2mL)by retrobulbar Tenon capsule perfusion. Bevacizumab concentrations in serum and vitreous were determined by double antibody sandwich Elisa at 1 and 3d after administration. The changes of bevacizumab concentrations in serum and vitreous of two groups were compared, and the fluorescence of retinal was observed by laser confocal microscope.

RESULTS: One day after administration, intravitreal concentrations of bevacizumab in vitreous of group A and B were 254.40±13.65 and 1.60±0.32μg/mL respectively. Concentrations of bevacizumab in serum of group A and B were 0.55±0.15 and 0.63±0.05μg/mL respectively. The changes of bevacizumab concentrations in serum between two groups did not vary significantly(t=1.168, P=0.277). At 3d after administration, concentrations of bevacizumab in vitre-ous of group A and B were 236.80±8.70 and 1.40±0.23μg/mL respectively. Concentrations of bevacizumab in serum of group A and B were 0.66±0.17μg/mL and 0.64±0.14μg/mL respectively. The changes of bevacizumab concentrations in serum between two groups did not vary significantly(t=0.207, P=0.841). For two administration routes, the fluorescence distribution of retina layers could be clearly detected.

CONCLUSION: At 1 and 3d after intravitreal injection, intravitreal concentrations of bevacizumab of group A is much higher than that of group B. Intravitreal injection of bevacizumab is the more effective route of administration for intraocular tissue. But retrobulbar Tenon capsule perfusion can also achieve the minimum concentration which can completely blocks VEGF activity(>500ng/mL), and can remain for at least three days. Both intravitreal injection and retrobulbar Tenon capsule perfusion of bevacizumab results in high serum concentration, and the change of bevacizumab concentration in serum between two groups does not vary significantly. In both administration routes, the fluorescence distribution of retina layers can be clearly detected, and both two dosing ways can work on each layer of retina.]]> 2015/8/27 11:19:42 Yi-Nan Wu,Hong-Jian Zhou,Guo-Hai Wu and Quan-Yong Yi 160true METHODS:One hundred and ten cases(110 eyes)with fungal keratitis from January 2012 to December 2014 were collected. The results of fungal smear, fungal culture and pathological examination results were analyzed retrospectively. Fungal smear was detected by 10% KOH wet microscopy and gram staining microscopy. Fungal culture was used potato dextrose agar(PDA)medium. The specimens of pathological examination were from corneal transplantation surgery. paraffin section, HE and hexamine silver and PAS staining was used in the pathological examination.

RESULTS:Of the 110 cases of fungal keratitis, fungal smear positive were observed in 50 cases(45.5%), fungal culture positive were observed in 55 cases(50.0%); pathological examination positive were observed in 88 cases(80.0%). Fifty cases were both fungal smear and pathological examination positive and 22 cases were both fungal smear and pathological examination negative. The coincidence rate of fungal smear and pathologic examination was 65.5%. Fifty-five cases were both fungal culture and pathological examination positive and 22 cases were both fungal culture and pathological examination negative. The coincidence rate of fungal culture and pathologic examination was 70.0%. In the 60 cases of fungal smear negative results, 38 cases(63.3%)were confirmed positive through pathological examination. In the 55 cases of fungus culture negative results, 33 cases(60.0%)were confirmed positive by pathological examination.

CONCLUSION:The accuracy of pathological examination is the highest. The combined application of fungal smear, fungal culture and pathological examination can improve the diagnostic accuracy of fungal keratitis.]]> 2015/8/5 15:43:41 Peng-Fei Chen,Xiao-Yi Qin,Li-Ping Mao,You-Pei Wang,Da-Xuan Wang and Mei-Qin Zheng 159true METHODS: Totally 30 Balb/c mice, aged 6～8wk, were randomly divided into the control group, light-exposure group and α-mangostin group. Every group contained 10 mice. Mice of α-mangostin group were treated with alpha-mangostin at the dose of 30mg/(kg·d)body weight by intragastric administration daily for 7d, and then exposed to white light at the 5^thd. The light-exposure group and α-mangostin group were exposed to 5 000±200lx white light-emmiting diodes(LEDs)for continuously 1h to establish the mice model of retinal light damage. Flash-electroretinograme was recorded 72h after light exposure. The changes in retinal morphology of mice were observed by light microscopy. Retinas were extracted to detect the malondialdhyde(MDA)content change of the retinal homogenate.

RESULTS: Flash-electroretinogram(F-ERG)showed that retinal dysfunction was less severe in α-mangostin group than in light-exposure group(P<0.05). Light microscopy test showed that retina structural damage was less severe in α-mangostin group than in light-exposure group(P<0.05). The level of MDA in retinal tissue of α-mangostin group was significantly lower when compared with light-exposure group(P<0.05).

CONCLUSION: α-mangostin inhibits lipid peroxidation induced by light damage and protect retina against light damage.]]> 2015/7/1 9:46:01 Yuan Fang,Tu Su,Ping Xie,Song-Tao Yuan,Wen Fan,Yi-Dan Xu,Zi-Zhong Hu and Qing-Huai Liu 158true 2O₂).

METHODS:Human RPE cells were subcultured, cell activity was detected by MTT, rate of apoptosis was detected by flow cytometry and cell ultrastructure changes were observed under transmission electron microscope.

RESULTS:MTT results showed that cell activity elevated to(53.66%±3.25% and 70.43%±2.38% after 10^-8mol/L and 10^-4mol/L AST treated. The difference had statistically significant(P<0.05)compared with oxidative injury group(38.76%±3.74%). Flow cytometry results showed that the apoptosis rate of RPE cells decreased to 30.23%±1.91% and 12.58%±2.12% in AST pretreated group, the difference was significant(P<0.05)compared with oxidative injury group(42.50%±1.94%); Electron microscopy showed that the morphology of cells gradually improved accompanied with the concentration of AST elevated.

CONCLUSION:AST may inhibit hydrogen peroxide-induced apoptosis of RPE cells, it can provide reliable evidence for pursue effective medicine to prevent and treat retina injury.]]> 2015/7/1 9:46:01 Hai-Rong Zhuang,Ping Liu and Xue-Zheng Hu 157true METHODS: The lens capsule or anterior capsular tissue of rat, rabbit, dog and patient were removed by different methods, and they were cut into tiny pieces for primary culture by modified tissue adherent method. The morphological features of four kinds of LECs were observed under an inverted microscope.

RESULTS: Four kinds of LECs of rat, rabbit, dog and human could be cultured primarily by tissue adherent method. With the evolution of tissue source, the adherent capacity of LECs gradually strengthened, cells form were changed from irregular polygon to oval, nucleus rounded and cytoplasm enriched gradually. Four kinds of LECs had fibrotic changes after several passages.

CONCLUSION: LECs of rat, rabbit, dog and human can be primarily cultured. This method lays the foundation for the mechanism research of caratact and related fields on the cellular and molecular levels.]]> 2015/7/1 9:46:01 Li-Xia Ji,Cai-Na Li,Quan Liu,Yi Huan,Shuai-Nan Liu and Zhu-Fang Shen 156true METHODS:Immunohistochemical technique was used to detect expressions of MMP-2 and EMMPRIN in 39 cases of paraffin embedded Rb samples. Quantitative analysis of expressions of MMP-2 and EMMPRIN were assessed by measuring the mean gray scale of Rb tissue with LEICA IM50 Color Pathologic Analysis System. The differences of expressions of MMP-2 and EMMPRIN in each clinical and pathological stage were statistically analyzed, and the same step was also undertaken to study the relationship between Rb with MMP-2 positive expression and that with EMMPRIN positive expression.

RESULTS: The positive expression rate of MMP-2 was 90%(Gray value: 109.64±14.52; 35/39), and that of EMMPRIN was 85%(Gray value: 108.01±13.60; 33/39). The expressions of MMP-2 and EMMPRIN were significantly higher in tumors of glaucomatous stage(Gray value: 108.21±11.47 and 107.56±14.32)than those in intraocular stage(Gray value: 121.13±11.32 and 119.34±12.66; P<0.01 and P<0.05). And the same conclusion can be concluded between those in extraocular stage(Gray value: 91.03±11.71 and 92.26±12.93)with those in glaucomatous stage(P<0.01 and P<0.05). The expressions of MMP-2 and EMMPRIN were significantly higher in tumors with optic nerve invasion(Gray value: 103.89±13.39 and 105.23±14.00)than those without optic nerve invasion(Gray value: 118.39±15.11 and 117.53±16.13)(P<0.01 and P<0.05).

CONCLUSION: The positive expression levels of MMP-2 and EMMPRIN may correlate with tumor infiltration and metastasis.]]> 2015/7/1 9:46:01 Yu-Hong Cheng,Qiang Shi,Jia-Quan Shen,Li-Lun Wang and Si-Wei Liu 155true METHODS:Twenty diabetic cataract patients, 20 age-related cataract patients and 20 traumatic cataract patients diagnosed from January 2012 to October 2014 in our hospital were selected. RT-PCR method was used to detect the content of SIRT1 gene in LECs of each group patients. Western blot method was used for the detection of SIRT1 protein content in lens epithelial cells, apoptosis rate of LECs was detected by TUNEL.

RESULTS: RT-PCR reaults showed that the relative content of SIRT1 mRNA in patients of traumatic cataract group was highest for 1.000±0.078, followed by the age related cataract group was 0.427±0.067, then diabetic cataract group was 0.389±0.112, those two groups compared with the traumatic cataract group, the difference was statistically significant(P<0.05); Western blot showed that SIRT1 protein expression in LECs of traumatic cataract patients was the highest, followed by the age related cataract group, diabetic cataract group the expression of SIRT1 protein was the minimum. The results of TUNEL showed that apoptosis rate of traumatic cataract group and age group LECs rates were(4.5±2.3)% and(8.7±4.1)%, respectively, the difference was not statistically significant; while the diabetic cataract group of LEC apoptosis rate was(24.3±6.1)%, by comparing traumatic cataract group and age related cataract group, the difference was statistical significance(P<0.05).

CONCLUSION: Expression of SIRT1 gene and protein decreased in LECs of diabetic cataract patients, suggesting that this gene was involved in diabetic cataract, this provides reliable theoretical basis for our further research in the future. Regulation of SIRT1 gene expression in LECs will explore the effective ways and provide a new idea for the diabetic cataract intervention treatment.]]> 2015/6/1 16:06:21 Shang-Li Jin,Hai-Ke Guo and Zhi-Hui Chen 154true METHODS: Sixteen paired rabbit lens were randomly divide into two groups, the lens of any pair of one was as experimental group, the other for the control group. Experimental group contained concentration of 5, 10, 15, 20, 25, 30, 35, 40mmol/L CaCl₂ 1 640 culture medium, the control group contained 1 640 culture medium, the situation was observed by turbidity after 36h. Twenty-four paired rabbits lens were randomly divided into two groups, the lens of any pair of one was as experimental group, the other for the control group. Experimental group contained CaCl₂(5～30mmol/L)+ E-64d of 1 640 culture medium and control group containd CaCl₂(5～30mmol/L)of 1 640 culture medium. Lens transparency and relative gray values were detected, Atomic absorption spectrophotometer was used to detect lens calcium(Ca^{2 +})content. The data were analyzed with SPSS 13.0 statistical software. Measurement data were expressed (-overx)±s and the differences of two groups were compared by pared- samples t test. P<0.05 was considered statistically significant.

RESULTS: Transparent lens opacity occurred in medium containing CaCl₂, and with the Ca²⁺ concentration increased, degree of lens opacity was also improved. When Ca²⁺ concentration >30mmol/L, black cross line below the lens was hardly seen through in culture medium, and lens cortex was almost completely cloudy. Lens opacity incubated with 1 640 mediun containing E-64d was declined compared with controls. There were significant difference of relative gray scale between experimental group and control group(t=3.820, P=0.001<0.01). However, experimental group and control group had no significant effect on Ca²⁺ uptake by lens(t=2.144, P=0.055>0.05).

CONCLUSION: The high levels of extralenticular calcium can induced cataract. E-64d, an inhibitor of calpain, can inhibit calcium-induced lens opacity, However, E-64d has no significant effect on Ca²⁺ uptake by lens. Its inhibitory effect on lens opacification may be due to a direct action on the activity of calpain. Moreover, maybe it can inhibit degeneration of crystallin or apoptosis and necrosis of epithelial cell by other approaches to delay the occurrence of cataract.]]> 2015/6/1 0:00:00 Yi Yang,Jian-Hua Lu,Wen-Fang Zhang and Dong-Mei Zhang 153true METHODS: RGC were cultured in 4 groups for 3d: control group, glucocorticoid group(with 0.1μmol/L cortisone), glucocorticoid-siNgR group \〖with 0.1μmol/L cortisone+Nogo receptor(NgR)antisense nucleotide\〗, glucocorticoid-scRNA group(with 0.1μmol/L cortisone + scrambled nucleotide). The cell viability was detected by thiazolyl blue tetrazolium bromide(MTT), the morphological features were observed with inverted microscope, apoptosis of RGC was measured with Hoechst 33342 staining, and expression of NgR was revealed by Western blot.

RESULTS: Cell viability in control, glucocorticoid, glucocorticoid-scRNA and glucocorticoid-siNgR groups were(100.0±0.0)%,(76.3±6.8)%,(79.4±9.0)% and(96.7±9.8)% respectively. Decreased cell viability, reduced cell number, and increased expression of NgR were atrophic cell body detected in glucocorticoid and glucocorticoid-scRNA groups(P<0.01), not in glucocorticoid-siNgR group(P>0.05), compared with control group. RGC was showed light blue by Hoechst 33 342 staining in control and glucocorticoid-siNgR groups, and exhibiting bright blued apoptotic RGC in glucocorticoid and glucocorticoid-scRNA groups.

CONCLUSION: Up-regulation of NgR contributes to glucocorticoids-induced apoptosis of RGC.]]> 2015/6/1 16:06:22 Li-Zheng Lü and Xue-Zheng Liu 152true Acin1(apoptotic chromatin condensation inducer 1)in congenital cataract mouse retina during development and investigate the differences of retinal apoptosis and the connection of lens and retina development between congenital cataract mouse and normal mouse.

METHODS: There were congenital cataract mice(10 female and 5 male)and normal C57BL/6 mice(10 female and 5 male). One male and two female mice were fed in the same cage randomly. The young mice were divided into two groups: congenital cataract group and normal control group. Five young mice were treated each group on 1, 5, 9, 14, 17, 21, 26, 60d. The left eyes were fixed with 4% neutral formalin to detect ACIN1 protein by immunohistochemistry and retinas from right eyes were used to detect the mRNA expression of Acin1.

RESULTS: Acin1 had sustained expression in each group. ACIN1 protein gradually expressed from the ganglion cell layer, inner nuclear layer to the outer nuclear layer following retinal development. It mainly expressed on ganglion cell layer and inner nuclear layer, but not neuroblastoma layer. ACIN1 protein positive cells on P1～P14d increased in normal control group, P17d reduced, after P21d positive cells of each layers decreased. The overall trend was similar in congenital cataract group with normal control group, P1～P14d positive cells count was lower than normal control group, P17-P21d positive cells were flat and higher than the normal control group. Compared with the same day of the two groups, the differences except for P17, P26, P60d were significant(P<0.05). The overall difference was statistically significant in congenital cataract group(F_cataract=295.07, P<0.01); in addition to P1 and P5, P17 and P21, the differences were statistically significant(P<0.05)compared with each other in congenital cataract group. The overall difference was statistically significant in control group(F_normal=214.21, P<0.01); in addition to P1 and P5d, the difference was statistically significant(P<0.05)compared with each other in control group. The expression of P17d in congenital cataract group was lower compared with that of P14d in control group, the difference was statistically significant(P<0.05). Acin1 mRNA trends of two groups were similar with ACIN1 protein. Compared with the same day of the two groups, the difference was significant except for P17, P21, P60d(P<0.05). The overall difference was statistically significant in each other of the two groups(F_cataract=522.29, P<0.01; F_normal=472.05, P<0.01). The difference was statistically significant compared with each day in control group(P<0.05). Compared with all the rest of days except for P21 and P26d, the difference was statistically significant in congenital cataract group(P<0.05).

CONCLUSION: Acin1 exist differential expression of time and space in mouse retina during development, congenital cataract crystal developmental disorder may affect the expression of Acin1 and retinal cell apoptosis and development.]]> 2015/5/5 16:34:50 De-Wei Li,Tao Jiang,Xiao-Yan Tong,Xiao-Chuan Wang and Shuang-Shuang Wang 151true METHODS: Seventy-two male Sprague-Dawley(SD)rats were chosen and divided randomly into 6 groups: normal control 2mo(C2m, n=12), diabetes mellitus 2mo(D2m, n=12), normal control 4mo(C4m, n=12), diabetes mellitus 4mo(D4m, n=12), normal control 6mo(C6m, n=12)and diabetes mellitus 6mo(D6m, n=12). The diabetes mellitus mouse were induced by intraperitoneal injection of 0.1mol/L streptozotocin(STZ, 65mg/kg). The expression of BIP, HIF-1α and VEGF in the retina were detected by enzyme-linked immuno sorbent assay. The location of BIP, HIF-1α and VEGF in epiretinal membranes were investigated by immunohistochemistry staining.

RESULTS: BIP were significantly increase than control groups in all DM groups with the course of diabetes(P<0.01). HIF-1α were detected higher than control groups in all DM groups(P< 0.05), but there was no statistical significance among treatment groups. VEGF in D4m and D6m groups were higher than control groups(P<0.05), and there was statistical significance between D4m and D6m groups. BIP can be detected in control groups a little, mainly in the inner limiting membrane and ganglion cell layers. HIF-1α cannot be detected in control groups, meanwhile it can be detected in all layers in DM. VEGF can be detected in control groups a little, it mainly be detected in inner nuclear layer, outer nuclear layer, around retinal vessel and ganglion cell layers in DM groups.

CONCLUSION:The expressions of BIP, HIF-1α and VEGF increase in the retina of diabetic rats than control groups with progressive of diabetes mellitus, both endoplasmic reticulum stress and HIF-1α signal path play important role in the progress of diabetic retinopathy.]]> 2015/5/5 16:34:51 Jing Wang,Hong Zhu and Cai-Hong Shi 150true METHODS: SD rats serum containing the prescription of reinforcing kidney, nourishing blood, improving eyesight and the content of distilled water in serum were prepared. The effects of the prescription and distilled water in serum at different concentration(2.5%, 5%, 10%, 20% and 40%)on cell vitality was observed by cell counting kit(CCK-8)assay. the logarithmic phase of ARPE-19 cells were pretreated by different concentrations(1.25%, 2.5% and 5%)of the prescription serum and distilled water in serum for 24h. Then it was treated with 75μmol/L acrolein for 24h. Cell vitality was observed by CCK-8 assay. The change of cell nucleus was detected by DAPI staining.

RESULTS: 2.5% and 5% serum had no effect on cell viability(P>0.05), while 10%, 20%, 40% serum could inhibit cell viability(P<0.05). CCK-8 results showed that 2.5% and 5% the prescription of reinforcing kidney, nourishing blood, improving eyesight serum was better than distilled water in serum(P<0.05).

CONCLUSION: The prescription of reinforcing kidney, nourishing blood, improving eyesight has the protective effect on ARPE-19 cell damage induced by acrolein.]]> 2015/5/5 16:34:52 Man Li,Li-Na Liang and Zeng-Yuan Zhuang 149true METHODS: Diabetes mellitus was induced by a high-fat and high-glucose diet in C57BL/6 mouse. 1% rose bengal was stained on the cornea to examine the integrality of the corneal epithelium at 2～12mo after completion of the model. Corneal epithelial wound healing was observed using a vivo epithelial debridement model which was dyed by sodium fluorescein. Corneal morphology histology was examined by pathological methods.

RESULTS: The high-fat and high-glucose diet C57BL/6 mouse in 2mo had showed general symptoms of diabetes: polydipsia, polyphagia, polyuria, weight loss etc. The model had a steady-state high glucose(≥18mmol/L), also the weight was lower compared with normal control mouse.1% rose bengal corneal staining had dot coloring at 2mo after completion of the model, the stained area and extent were gradually increased with the extension of the duration of diabetes, almost all the cornea was stained at 12mo after completion of the model. With the passage of time into a mold, the cornea epithelial healing time become longer: 2mo was about 40h; 3mo was about 120h; 4, 6, 12mo was about 144h; the coloboma were gradually increased at 12mo after completion of the model, then the area was reduced gradually until complete healing, the time was 96～120h, showed repeating phenomenon.

CONCLUSION: The mouse were induced by high-fat and high-glucose diet can be used as animal models of diabetic keratopathy: the damage of epithelium for corneal and delay healing on epithelium and other symptoms.]]> 2015/4/8 9:56:56 Wen Bo,Guang-Hua Sun,Feng-Xia Sun and Wen Cui 148true METHODS:Thirty-two albino rabbits were randomly divided into four groups, including standard control group, experimental group Ⅰ, experimental group Ⅱ, and experimental group Ⅲ. Each eye was performed standard trabeculectomy. During surgery, standard control group was given a piece of cotton with 0.3mg/mL mitomycin C(MMC)for 2min, and the other three groups were given a piece of cotton with 0.3, 0.4, and 0.5mg/mL suramin respectively for 2min. The filtering blebs and intraocular pressure(IOP)were observed at the 3, 7, 15, and 30d after surgery. Some conjunctiral specimen were observed with hitochemicall(HE staining)and immunohistochemicall methods.

RESULTS: At postoperative 7, 15, and 30d, the changes of the IOP, functional filtering blebs, and the number of positive cell nuclear in experimental group Ⅱ and experimental group Ⅲ were significantly different compared with those in standard control group and experimental group Ⅰ(all P<0.05), and the differences between experimental group Ⅰ and standard control group were not significant(P>0.05). The changes of the IOP and the number of positive cell nuclear in experimental group Ⅲ were significantly different compared with those in experimental group Ⅱ(P<0.05), whereas the differences in functional filtering blebs between experimental group Ⅲ and experimental group Ⅱ were not significant(P>0.05). The status of filtering channel in experimental group Ⅱ and experimental group Ⅲ were better than those in experimental group Ⅰ and standard control group.

CONCLUSION: The concentration of suramin has a significantly influence on its effect. When the concentration is 0.3mg/mL, the antiproliferative effect of suramin is no more than that of MMC. The effect of 0.4,0.5mg/mL suramin is better than MMC. 0.5mg/mL suramin has a better effect on controlling IOP and suppressing the growth of fibroblasts than 0.4mg/mL suramin.]]> 2015/4/8 9:56:56 Bo Yao and Zhi-Kun Xin 147true METHODS: Neonates of C57BL/6 mouse(P7)were exposed to 75%±2% oxygen for 5d and return to normal air environment when 12d(P12)to establish oxygen-induced retinal neovascularization model. The neonates were divided into groups, injected with 0.5μL solution containing 25ng(group A), 50ng(group B), 250ng(group C)of soluble erythropoietin receptor(EPO-R)or PBS(group D)at P12, P14 and P16 in the right eye. On P17, the litters were sacrificed and their right eyes were enucleated and fixed with 4% paraformaldehyde, made pathological section. The number of breakthrough internal limiting membrane neovascular nuclei was counted with pathological retinal morphology, understanding theproliferative degree of retinal neovascularization.

RESULTS: The pathological sections showed the neovascular cell nuclei which penetrating the inner limiting membrane in intravitreal EPO receptor injection group was reduced statistically than that in PBS injection group, the difference was statistically significant(P<0.01). And, neovascular nuclei count differences in the various concentrations of EPO receptor group was statistically significant(P<0.01). With the EPO receptor concentration increase, neovascular endothelial cells broken through the internal limiting membrane was reduced.

CONCLUSION: Intravitreal injection of soluble EPO receptor can block EPO and improve neovascularization. The new method is expected to become new treatment of ocular neovascular diseases.]]> 2015/4/8 9:56:56 Wen-Zhi Huang,Qian-Qing Li,Lu Wang and Wei Zhang 146true METHODS: Forty mature healthy female SD rats were randomly divided into study group and control group, 20 rats in each group, the study group were ovariectomized, low concentration of naphthalene long interval of administration, the establishment of perimenopausal ARC model, the control group of conventional farming. In the subtractive hybridization cloning by MRG15, and make the probe of the cDNA fragment using digoxigenin labeled. Access to the two groups of rats anterior lens capsule after slicing, and then through the differential expression in situ hybridization clear lens epithelial cells.

RESULTS: The cloned MRG15 through BamH1, EcoR1 enzyme digestion and agarose gel electrophoresis, available for 639bp long cDNA fragment. GeneBank display contrast, their homology was 99.0%. In situ hybridization, ARC patients and normal lens epithelial cells were observed in the expression of MRG15. Study group the percentage of positive cells compared with control group, showed a significant difference(P<0.05). Integral index study group and the control group compared with, was significantly difference(P<0.05).

CONCLUSION: ARC, MRG15 in normal lens epithelial cells expressed ARC, and compared with the normal expression of lens epithelial cells, which may produce inhibitory effects are associated with MRG15 transcription in human lens epithelial cells in the part of key genes, by reducing the lens epithelial cell function, make its appear aging, and the formation of cataract, clinical response to this should further study.]]> 2015/3/9 10:14:41 San-Quan Jin 145true METHODS: Twenty samples of normal conjunctiva and the aggressive and quiescent period of pterygium of both 50 cases were studied under the light microscope by HE stain, Periodic acid-Schiff stain(PAS), immunohistochemistry staining with CD34, VEGF and immunohistochemistry combined with PAS stain.

RESULTS: The expression of CD34 and VEGF in pterygium was significantly higher than those of normal conjunctiva specimens(All P<0.05). The expression of CD34 and VEGF in aggressive cases was apparently higher than that in quiescent cases(All P<0.05). Vasculogenic mimicry was identified in 38 of 50 aggressivcases and 11 of 50 quiescent cases by PAS stain, with a statistically significant difference(P<0.05). Bivariate analysis showed that vasculogenic mimicry were positively correlated with aggressive cases(Spearman's correlation coefficient r=0.540>0.5, P=0.000).

CONCLIUSION: Neovascularization exists in pterygium and vascular mimicry is one of the blood supply pathways of the pterygium and play an very important role in the development of pterygium.]]> 2015/3/9 10:14:41 Jun-Jie Chen,Yu-Qing Lan,Gong-Fa Wu,Jun-Shan Lin,Rui-Zhang Ou and Yu-Ting Zeng 144true METHODS: DR rats were fed with Shuangdanmingmu capsule. By comparing with the normal group, the model control group, and positive control group, the effect of Shuangdanmingmu capsule on retinal tissue of DR rats was observed under electron microscopy. After HE staining, retinal structure was observed under the light microscope. Immunohitochemical staining was used to detect the VEGF expression in retina.

RESULTS:Two months after treatment, the layers tissue of retina presented mild edema, capillary pericytes performed edema, mitochondria showed mild swelling and less clear structure, some endothelial cells showed slight proliferation in Shuangdanmingmu group. Compared with the normal group, the expression level of VEGF in retina increased in the other groups, especially in model control group. A significant differential in expression of VEGF was found between Shuangdanmingmu group, positive control group and model control group(P<0.01).

CONCLUSION: Shuangdanmingmu capsule can effectively improve the retinal microvascular, reduce edema and necrosis of each layer of retina, improve the ultrastructure of retina's tissue and inhibit VEGF expression in DR rats.]]> 2014/12/30 15:26:04 Yu-Hui Qin,Wen-Juan Li,Xi Zhang,Zong-Shun Dai,Xiao-Liu Chen,Ya-Sha Zhou,Yan-Jun Ling and Bing Zheng 143true METHODS:Research group were dealt for thermal transient receptor channel 1 gene mediated by liposome transfection to rabbit corneal endothelial cells. MTT method was used to observe its influence on cell proliferation. Immunohistochemical staining and computer image analysis system were used to test the effects for proliferation cell nucleus antigen(PCNA)expression.

RESULTS:Proliferation of corneal endothelial cell of rabbit was promoted after thermal transient receptor channel 1 gene transfected and the difference between experiment group and control group(t=3.01,P=0.013). The expression of PCNA promoted after thermal transient receptor channel 1 gene transfected(t=3.21,P=0.007)compared with control group.

CONCLUSION:The expression of PCNA in rabbit corneal endothelial cells can promote the proliferation of corneal endothelial cells of rabbits.]]> 2015/11/27 10:32:21 Li Wang,Zhao-Jiang Du and Peng Li 142true METHODS:Patients suffering from B-cell non-Hodgkin lymphoma(NHL)(n=40)and reactive lymphoid tissue hyperplasia(RLH)(n=10)of ocular adnexal from June 1995 to June 2015 in Qingdao Affiliated Hospital were observed and the 50 paraffin sections were collected. RLH sections were selected to the control group. The patients' age, gender, pathogenic site and pathological types were selected as the classification criteria. The expression of Livin and Caspase-3 proteins were detected by immunohistochemistry and the positive expression rate between lymphomas and RLH was compared. Spearman rank correlation was used to estimate the relationbetween Livin and Caspase-3 in ocular adnexal lymphoma.

RESULTS:The expression of Caspase-3 in B-cell NHL was lower than that in lymphadenosis(P<0.05). While the expression of Livin in NHL was higher(P<0.05). The two proteins had no relation with the age, pathogenic site or gender. But they were related to pathologic type. Livin was highly increased in plasmacytoma(PL)and diffused large B-cell lymphoma(DLBCL)compared with mucosa-associated lymphoid tissue(MALT)lymphoma(P<0.05). While Caspase-3 in PL and DLBCL were lower than that in MALT lymphoma(P<0.05). There was a negative correlation between Livin and Caspase-3 in MALT(r=-0.491,χ²=7.519,P<0.05).

CONCLUSION:Over expression of Livin and low expression of Caspase-3 may play a significant role in the occurrence, development and different pathologic type of ocular lymphoma. The expression of Livin and Caspase-3 combined with each other in NHL. Combined examination of two proteins may be a valuable marker to predict the presence and differentiate the pathologic type of ocular adnexal lymphomas.]]> 2015/11/27 10:32:21 Jun-Xia Lu,Hong Lin,Fen Wang,Jing Lin,Li-Na Zhang and Gui-Qiu Zhao 141true METHODS:Sprague-Dawley(SD)rats were divided into normal group(group a,n=12)and model group(n=24). Rats in model group received intraperitoneal injection of streptozotocin(60mg/kg body weight)to get diabetic rats,which were randomly subdivided into diabetic group(group B,n=12)and xueshuantong intervention group \〖group C,1g/(kg·d),n=12\〗. At 6th and 12th wk of the experiment, 6 rats randomly selected from every group,were executed to get retinal tissue for detecting CTGF-mRNA by electron microscope.

RESULTS:Compared with group A,the content of CTGF-mRNA in retina tissue increased at 6, 12wk in both group B and C(P<0.05). Compared with group B,the content of CTGF-mRNA in retina tissue in group C decreased at 6, 12wk(P<0.05). Electron microscope showed:in group B,the capillary endothelial cells and mitochondria became swollen; the amount of pinocytosis vesicle increased; the electron density in capillary basement membrane was not homogeneous and nodular thickening phenomenon appeared; compared with group B,the changes in group C were alleviated.

CONCLUSION:CTGF may be involved in the progress of DR in diabetic rats. Compound xueshuantong may exert its protective effect through decreasing the expression of CTGF in the retina of diabetic rats.]]> 2015/11/27 10:32:22 Yu-Wei Xing,Yang Yang and Xue-Jiao Chai 140true METHODS: The ERG of 50 mice(50 eyes)of KUNMING at the ages of postnatal 14d(P14), P21, P28, P35 and P56 were measured respectively. The implicit times and amplitudes of b wave of Rod-ERG, a and b waves of Max-ERG, a and b waves of Cone-ERG and O1 and O2 waves of OPs at different ages, as well as amplitude of Flick-ERG, were compared.

RESULTS: The Max-ERG a-waves(the 95%CI were 15.00～18.60, 12.00～15.00, 13.20～14.40, 13.20～15.00, 13.20～15.00, respectively), OPs O1(the 95%CI were 15.00～19.80, 13.80～18.00, 13.20～14.40, 13.80～15.60, 13.80～15.60, respectively)waves shared the implicit times at the different stages, and the Flick-ERG(the 95%CI were 0.97～3.28, 0.85～2.32, 0.91～3.49, 0.94～2.68, 0.98～3.69μV, respectively)shared the amplitudes also. There was no significant difference among the weeks(P>0.05). The implicit times of the Cone-ERG a-waves(the 95%CI were 25.20～55.20, 27.00～40.20, 27.00～38.40, 25.20～43.80, 23.40～37.80, respectively)between P14 and P28 were distinct with statistical difference(P<0.05). The implicit times of Cone-ERG b-waves(the 95%CI were 70.80～88.20, 56.40～78.60, 60.00～75.60, 60.60～87.00, 62.40～81.60ms, respectively)at P14 were statistically different from those at P21 and at P28. The implicit times and amplitudes of Rod-ERG b-waves(the 95%CI were 87.00～114.00, 53.40～73.80, 52.2～63.6, 55.20～71.40, 57.60～67.80ms, and 64.21～195.07, 133.79～355.71, 130.62～355.96, 190.92～448.97, 239.26～462.40μV, respectively), Max-ERG b-waves(the 95%CI were 67.20～107.40, 32.40～54.60, 31.20～36.60, 31.80～42.00, 34.20～41.40ms, and 160.64～344.48, 281.74～590.09, 284.91～716.80, 358.64～737.55, 406.98～810.55μV, respectively), and OPs O2 waves(the 95%CI were 49.8～69.6, 29.40～42.60, 28.80～33.60, 28.80～37.80, 31.20～37.20ms, and 5.43～24.84, 54.38～147.52, 65.55～201.60, 46.33～164.79, 49.07～148.32μV, respectively)at P14 were different from those at other stages, and the amplitudes of OPs O1(the 95%CI were 11.60～21.36, 6.77～53.71, 32.96～76.42, 34.06～70.37, 35.58～63.35μV, respectively)and Cone-ERG b-waves(the 95%CI were 5.10～15.85, 9.61～24.88, 14.96～40.73, 14.87～28.54, 13.83～51.98μV, respectively)were from those at other stages also, and there were significant differences. The O1 wave of OPs had been present at P14, but the second cluster of OPs of one mouse(1/10)had not been obvious at the same time.

CONCLUSION: The experiment confirms that the different waves come from different cells in retina in mice at certain degree. Due to the change of the ERG in the development of mice, so it should be considered that the different development stages will affect the results when measuring the ERG of mice.]]> 2015/10/30 17:17:32 Hou-Cheng Liang,Ting Ma,Tan Long and Hong-Bing Zhang 139true METHODS:Group A: anterior chamber maintainer and supplementary lens were applied in anterior-posterior combined surgery. Group B: conventional anterior-posterior combined surgery was used. The application value of this device were analyzed from the operation time, operation effect, postoperative complications.

RESULTS:The application of anterior chamber maintainer with supplementary lens in anterior-posterior combined surgery played a positive role on the stripping time(P<0.05), but there was no significant difference on operation effect and postoperative complications(P>0.05).

CONCLUSION:There are advantages of anterior chamber maintainer with supplementary lens in anterior-posterior combined surgery, such as more convenient in operation process, a wider range of observation, bimanualness, less complications and so on.]]> 2015/10/30 17:17:32 Guang-Sheng Chen,Wei-Na Li,Hong-Bo Huang,Sheng Yang,Xin-Zhu Gan and Jun Fan 138true METHODS:Fourty five healthy new zealand rabbits were divided randomly to three groups:normal group, control group and experimental group. The control group was just performed vitrectomy with BSS; the experimental group was performed vitrectomy combined with nonexpansile perfluoropropane gas. The lens were made into tissue homogenates to detect T-SOD activity and MDA content by using spectrophotometry on the 1st,3th,8th,35th and 45th day after the operation.

RESULTS:T-SOD activity: that in normal group was the hightest, that in experimental group was the lowest, and there was significant difference(P<0.01)among three groups in every periods after operation. MDA content: that in normal group was the lowest, that in experimental group was the hightest, and there was significant difference(P<0.01)among three groups after the operation except the 1st day.

CONCLUSION:Oxidative damage mechanism may be involved in the early damge of rabbit lens after vitrectomy combined with nonexpansile perfluoropropane gas.]]> 2015/10/30 17:17:33 Yan-Fang Yang,Song-Bo Jia and Luo-Sheng Tang 137true METHODS: Using pSilencer2.1-U6neo for plasmid vector, HIF-1α siRNA recombinant plasmid was constructed. There was totally 54 healthy Sprague Dawley rats in which 15 rats were chosen as normal group and 39 rats were constructed for diabetic retinopathy model by streptozotocin(STZ)which was divided into three subgroups randomly including control model group(DR group, 15 rats), vector group(12 rats)and gene therapy group(HIF-1α siRNA group, 12 rats). Nothing was transfected into DR group and normal group. The vector plasmid and HIF-1α siRNA were injected into the vitreous in vector group and HiF-1α siRNA group respectively. The retinal morphology was observed by hematoxylin-eosin(HE)staining and the expression of VEGF protein was measured by immunohistochemical staining. The inhibition efficiency of VEGF was calculated at 24, 48, 72h and 1wk after injected. Significant differences between groups were evaluated by one-way analysis of variance, followed by LSD-t analysis.

RESULTS: HIF-1α siRNA recombinant plasmid was confirmed by enzyme digestion and sequence analysis. HE staining showed that the retinal cells at each layers in normal control group were arranged regularly, and cell's morphology was roughly normal. The retinal cells at each layers arranged in disorder in diabetic rat And the inner limiting membrane was not complete with neovascular buds and neovascularization cluster growing out of the inner limiting membrane vertically. Immunohistochemical staining showed that the positive expression of VEGF was brown yellow granules, which was mainly located in ganglion cell layer. It also revealed the expression of VEGF protein was weakly positive in normal control group, while the DR group and empty vector group were significantly increased. Compared with DR group and the empty vector group, gene therapy group was significantly decreased, the difference was statistically significant(P<0.05). VEGF protein level was reduced by 27.4%, 40.6%, 47.5%, 64.5% at 24, 48, 72h and 1wk.

CONCLUSION: Intravitreal injection with HIF-1α siRNA can efficiently inhibite VEGF protein in retina of diabetic rats, which may be a new method for the treatment of diabetic neovascular disease.]]> 2015/9/25 17:05:18 Li-Zhu Meng,Song Chen,Lei Chen,Yan Liu,Jin-Yong Lin,Yu-Chuan Wang and Mei Han 136true METHODS: New Zealand rabbits with dry eye was prepared and treated with 0.1% and 0.3% sodium hyaluronate drops fluid respectively, which were regarded as low concentration treatment group(group B)and high concentration treatment group(group C)respectively. However, the rabbits treated with saline were regarded as control group(group A). And then, corneal fluorescein staining, Schirmer test, conjunctival goblet cells, mucin expression and histological changes were observed.

RESULTS: On D7 and D14 after treatment, corneal fluorescein staining scores were lower in group B and group C than that in group A(P<0.05). However, Schirmer test, goblet cell density and mucin content were higher in group B and group C than those in A group(P <0.05). Tear secretion and goblet cell density were higher in Group C than those in group B(P<0.05). Compared with group B and group C, the thicknesses of corneal and conjunctival epithelial cell layer were thinner in group A. There were not obvious abnormalities in corneal and conjunctival stroma in each group.

CONCLUSION: The sodium hyaluronate can improve ocular surface damage of dry eye in New Zealand rabbits. The high concentration of sodium hyaluronate has better effect than low concentration.]]> 2015/9/25 17:05:18 Shuang-Yong Wang,Ying Tian,Yan Cheng,Hai-Feng Zhu and Jie Wu 135true METHODS: The high glucose-induced HLEC model was established. Different concentrations of ginkgolide B were intervened. Cell viability was assayed by MTT assay, morphology of cell apoptosis was observed by hochest33258 staining. Cell ultrastructural changes were detected by transmission electron microscopy. Expressions of apoptosis-related factors caspses-3 and caspase-9 were tested by colorimetric detection.

RESULTS: High glucose inhibited the activity of HLECs, induced apoptosis reaction of HLECs, caused high expression of apoptosis factors in cells; while ginkgolide B inhibited the decrease of cell viability induced by high glucose, decreased the HLEC apoptosis and reduced the expression of apoptosis factors Caspase-3 and Caspase-9.

CONCLUSION: Ginkgolide B may inhibit the expression of caspses-3 and caspase-9 then effectively inhibited high glucose-induced apoptosis in HLECs.]]> 2016/6/29 17:05:28 Yan-Hong Wang, Tian-Ye Lan, Ting Chen, Ping Liu and Li-Li Lin 134true METHODS: Sodium iodate at 35mg/kg(body weight)was administered by tail vein injection into adult 6-8wk C57BL/6J mice. Morphological and functional changes of the retina were assessed at 6h, 1, 3, 5 and 8d after injections by fundus imaging, optical coherence tomography(OCT), ERG and histology. Mice in control group were give tail vein injection of equivalent dose of normal saline. All the eyeballs were removed for paraffin section and H-E staining.

RESULTS: The fundus photographs images at 6h after injection showed obvious changes, which were light red in retina and showed retinal blood vessels radial arranged. At 6h after injection, off-white drusen-like change was found at fundus. While there were no observable changes in OCT image and ERG. At 1d after injection, the fundus lesion aggravated and the drusen increased gradually. There were retinal pigment epithelial(RPE)disorders, photoreceptors and outer nuclear layer(ONL)damage through OCT. At 3d after injection, the retina lesion aggravated further and the retina became edema. At 5d after injection, the retina edema cleared away, the optic nerve became white and the fundus lesions increased. At 8d after injection, the RPE layer, photoreceptors and ONL were destroyed obviously. In the process, ERG showed the amplitudes of a- and b-wave decreased in a time-dependent manner. H-E staining showed that cells in retina of mice in control group were neatly arranged and well-distributed. The outer layer retina of sodium iodate injection group was wave-like, the normal structure of RPE disappeared and black round sediment could be seen which increased with time. At 8d after injection, there were any normal RPE.

CONCLUSION: The tail vein injection of sodium iodate can well simulate the pathogenesis of age-related macular degeneration which can provide a good animal model for AMD.]]> 2016/5/31 16:15:54 Shuang Jiang and Hai-Yue Xu 133true METHODS:Fifteen wild type mice were used as the control group, and 10 Eaf2 KO mice were used as the experimental group. The 14-week mice were taken as the research objects in the two groups. So the subgroups were: WT -nonUV, WT -UV, Eaf2 KO-nonUV and Eaf2 KO-UV, a total of 4 groups. Observe the lens of mice in vivo with slit lamp microscope, grade the lens opacity with Lens Opacities Classification System II(LOCSII). Then the mice were sacrificed by breaking the neck, the lens were removed and were observed by dark field microscopy. According to the captured images, the proportion of cataract region was analyzed using Image J software. The data of the two groups were statistically analyzed.

RESULTS: The results detected by the two methods were similar. In WT-UV group and Eaf2 KO-UV group, the degree of lens opacity was significantly higher than those of WT-nonUV group and Eaf2 KO-nonUV group. The lens opacity of WT-UV group was significantly higher than that in Eaf2 KO-UV group, and the difference was statistically significant(P<0.05).

CONCLUSION: Ultraviolet radiation can lead to the formation of cataract in mice. Eaf2 protein can promote the formation of cataract in mice caused by ultraviolet.]]> 2016/2/3 8:48:35 Yan-Hua Jiang and Jin-Song Zhang 132true METHODS: Twenty-four adult rabbits were divided randomly into normal control group, model group and curcumin group. Except normal control group, the rabbit model with acute ocular hypertension was established by increasing intraocular pressure through anterior chamber infusion. After the establishment of the model, rabbits in curcumin group were injected intravitreally with curcumin(0.1mg/0.1mL)for 7d. Rabbits in mdodel group were injected with physiological saline replaced of the same volume curcumin. The oculars in normal control group without any treatments were acquired directly. The levels of Thy-1 in RGCs were detected by immunohistochemistry. The density of RGCs was counted too.

RESULTS: Thy-1 expressed in the RGCs in normal control group and curcumin group had no significant difference(P>0.05).While the difference between model group and normal control group on Thy-1 expression was statistically significant(P<0.05). The density of RGCs in curcumin group, normal control group and model group were 20.3±2.7, 21.5±1.8 and 15.1±2.3 cells/HP.

CONCLUSION: In this study on curcumin in adult rabbits with acute ocular hypertension, curcumin can increase the expression of Thy-1, which shows that it can partly reduce the injury of RGCs in rabbits with acute intraocular hypertension and curcumin may be able to protect the retina under certain situation.]]> 2016/2/3 8:48:35 Zhi-Gang Xu,Shu-Hui Lü,Yu-Qing Wang,Xiao-Tian Yang and Zi-Rui Liu 131true METHODS:The glaucoma rat models were made by cauterization of three episcleral veins. Then the glaucoma rats were divided into three groups. Group 1 was the untreated intraocular hypertension group. Group 2 was the low dose of chrysophanol group(25mg/kg). Group 3 was the high dose of chrysophanol group(50mg/kg), 15 rats in each group. The right eyes were the experiment eyes while the left were the control ones. After 6wk treatment, the mRNA and protein of protein kinase-like endoplasmic reticulum kinase(PERK)and Rho kinase 1(ROCK-1)were determined in the retina.

RESULTS:The chrysophanol reduced intraocular pressure(IOP)of experiment eyes, which was significantly lower than that of control eyes(P<0.01). Compared with the normal group, the p-PERK protein increased significantly in the retina of glaucoma model group and chrysophanol increased the lever of p-PERK protein. The ROCK-1 protein level increased significantly in glaucoma group, it all decreased in these treatment groups, and it decreased significantly in high dose treatment group. Detected by TR-PCR, chrysophanol also could activate the mRNA of PERK and inhibited the mRNA expression of ROCK-1 in a rat model of glaucoma.

CONCLUSION:These results suggest that chrysophanol can reduce the IOP through the phosphorylation of PERK protein to regulate the PERK/ROCK signaling in glaucoma rat model.]]> 2015/12/28 16:15:58 Hao Hu and Ling-Li Jiang 130true 2O₂).

METHODS:ARPE-19 cells were cultured conventionally and divided into four groups. One group was untreated as blank group, the other three groups were incubated in 75μm/L H₂O₂, 150ng/mLBMP-6 or75μm/L H₂O₂+150ng/mL BMP-6. All the groups were incubated for 3h, 6h, 9h and 12h. We tested the cell viabilitity by MTT. We used flow cytometry to test the cell cycle and cell apoptosis.

RESULTS:H₂O₂ significantly decreased the cell activity in time-dependent manner. The activity of cells with BMP-6+H₂O₂ was higher H₂O₂ group, and the differences between the two groups at 3h and 6h were significant(P<0.05). The observation on cellular morphology showed that the cell number decreased and the cell detachment after 6h incubated with H₂O₂, while the cells with BMP-6 were less cell detachment and apoptosis.

CONCLUSION:BMP-6 has protective effects on RPE cells from oxidative stress in certain extent.]]> 2015/12/28 16:15:58 Li Chen,Ming Liu,Yong Liu and De-Xiu Zhang 129true METHODS:OIR model was induced by exposure of mice from groups A and B(the retinopathy was known to be more severe in group B than that in group A by previous research)to high oxygen(75%)from postnatal day 7(P7)to P12(n=12 for each group)and returned to normal environment at P12. On P17, the mice from both groups were randomly assigned to accept FFA or high molecular weight fluorescein isothiocyanate dextran(FITC-Dextran)perfusion combined with stretched preparation of retina(each method involved 6 pups from each group). The retinal non-perfused areas were quantified and compared by using image analysis software.

RESULTS: FFA combined with image analysis software was able to quantify the retinal non-perfused areas, which was comparable to the results analyzed by FITC-Dextran perfusion combined with stretched preparation of retina. No statistical difference was found between the results obtained by the two methods(P>0.05).

CONCLUSION:FFA combined with image quantification analysis can be used in retinalvasculopathy evaluation in mouse OIR model.]]> 2015/12/28 16:15:59 Rong Li and Yuan Chang 128true METHODS: The data in this study is from Gene Expression Omnibus(GEO)which belong to Nation Center for Biotechnology Information(NCBI), the quality of the raw data CEL files was processed and analyzed by the Expression software which belong to Affymetrix Inc., Santa Clare, CA, USA. Significant analysis method(SAM)which base on the T test was used to identified the significant genes. Based on GRNInfer and Gvedit soft we set up gene networks of optic and retina of mice and further more enriched analysis which based on DAVID and MAS3.0 online software were processed.

RESULTS: The analysis between the group of the optic nerve heads and retinas in different stage of glaucoma showed that the amount of significant different expressed genes in the optic never head group increased significantly comparing with the group of retina in the early stage of glaucoma, the analysis of the genes network construction show that: the node genes of optic nerve heads included Unc13c、Kif5a、TRPM1、PANX; and the node genes of retina include POU4F1, NEFL, BC03870, CALB2. Metabolic pathways enrichment analysis which based on MAS3.0 online platform show that there was mainly the amyotrophic lateral sclerosis, tyrosine metabolism, melanogenesis, Nitrogen metabolism, Gap junction, Leukocyte transendothelial migration metabolism pathway enriched out in optic nerve head; and there was mainly amyotrophic lateral sclerosis, neurodegenerative disorders, prostate cancer, leukocyte transendothelial migration metabolism pathway enriched out in retina.

CONCLUSION: By understanding bioinformatics result, it seems optic were more sensitive than the retina to high intraocular pressure, and weather high expression of TYrp1 gene can be as a sensitive diagnostic item require more evidence back up. Functional enrich analysis of node gene showed that cytoskeleton reconstructed,molecular motor and nutrients transport function improve in optic; and in retina, the most prominent finding in retina was enrichment function modules were focus on regeneration, repairing and differentiation of cells, which remind that we should reinforce research on reparation of retina of primary glaucoma. Metabolic pathways enrichment analysis show that inflammatory response plays prominent place in optic and retina of primary glaucoma, because of the optic narrow and crowed anatomic shape, nutrient metabolism and substances transfer enrichment modules play an important role in optics of primary glaucoma.]]> 2016/10/25 14:35:36 Jing-Kun Liu, Lin-Bang Wang, Bing Wang, Ya-Ling Sheng, Jing He and Fen-Ge Meng 127true METHODS: Rabbit cornea alkali-burned model was made, then 50 rabbits were randomly divided into the experimental group(n=25)and the 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, and took photos in two groups. The infiltration of PMNs was identified by hematoxylin eosin(HE)staining in different periods.

RESULTS:The quantity of PMNs increased on the 3d, reached the lower level on 7d, shown a peak on the 14d, then decreased gradually. PMNs level of the experimental group was significantly lower than that in the control group, and the difference of 3, 14 and 21d was significant(P<0.05).

CONCLUSION: During the wound healing process, alkali-burned cornea has close relation with the infiltration of PMNs. The treatment of conjunctival flap covering for the severe alkali-burned cornea was found to have good effect.]]> 2016/9/19 17:26:48 Dong-Yu Song, Ming-Hong Gao and Shan-Shan Cui 126true METHODS: Totally 40 male SD rats were randomly divided into 6 groups, that was: A: blank group, B: model group, C: Qingguang'an II low dose group, D: Qingguang'an II moderate dose group, E: Qingguang'an II high dose group, F: Yimaikang disket group. B, C, D, E, F groups of experimental rats were established the model of chronic high intraocular pressure(IOP)by cauterizing of superficial scleral vein. Animal model was established successfully by using monitoring IOP consistently keep above 25mmHg for 8wk as cut-off criterion. Tissues of Eyes were obtained after intragastric administration for 2wk and 4wk. The expressions of PAX6, Ngn1, and Ngn2 mRNA were investigated by Real-time PCR.

RESULTS: At the time-point of 2wk, PAX6, Ngn1, and Ngn2 mRNA in group B were statistically expressed in lower level comparing with other groups(P<0.05). Moreover, at the time-point of 4wk, PAX6, Ngn1, and Ngn2 mRNA in group C, D and E were statistically expressed in higher level comparing with group F(P<0.05). Besides, PAX6, Ngn1, and Ngn2 mRNA in group C and D were statistically expressed in lower level comparing with group E(P<0.05). PAX6, Ngn1, and Ngn2 mRNA in group C and D were expressed in similar level(P>0.05).

CONCLUSION: In summar, Qingguang'an II and Yimaikang disket can remarkably increase the expressions of PAX6, Ngn1, and Ngn2, which suggest protecting the optic nerve of rats caused by chronic high IOP. What's more, this study indicated that, in the protection of optic nerve of rats with chronic high IOP, the high dose of Qingguang'an II at the time-point of 4wk was the better choice.]]> 2017/8/22 10:31:08 Ya-Sha Zhou, Jian Xu, Yue Liu, Jun Peng, Yi-Jing Yang, Gen-Yan Qin and Qing-Hua Peng 125true METHODS: Establishment of rabbit glaucoma ischemia reperfusion model. Twenty-four New Zealand white rabbits were randomly divided into three groups: nerve growth factor group, Ginkgo biloba extract group and combination group. Respectively, in the continuous administration of 1, 7, 14d. We observed the morphological changes of the tissues of the retina. The levels of superoxide dismutase(SOD), nitric oxide(NO)and malondialdehyde(MDA)in retinal tissue were measured.

RESULTS: Respectively, first, in the continuous administration of 1, 7, 14d, the contents of MDA and NO in Ginkgo biloba extract group and nerve growth group were higher than that in combination group(P<0.05). Secondly, the SOD content of Ginkgo biloba extract group and nerve growth group were lower than that of combination group at each time point(P<0.05). At each time point, the number of HE staining of retinal ganglion cells(RGCs)showed that the loss of RGCs in the combination group was significantly lower than that in the other groups, and the ganglion cell count showed that the Ginkgo biloba extract group and the neuronal growth group were lower(P<0.05).

CONCLUSION: Nerve growth factor combined with Ginkgo biloba extract has better protective effect on retinal ischemia-reperfusion injury. The mechanism may be related to the decrease of free radicals and increase the activity of SOD in retinal tissue.]]> 2017/8/22 10:31:08 Yue-Mei Li, Qing-He Li and Xin-Hua Zheng 124true METHODS: HTFs were cultured and identified by vimentin staining with immunofluorescence and the morphological characteristics. The experimental group was processed 48h with LiCl in concentration of 80mmol/L, the control group without LiCl. The mRNA expression of TGF-β and CTGF in two groups were analyzed with real-time fluorescent quantitative polymerase chain reaction(real time-qPCR)and the protein expression was detected with Western blot.

RESULTS:The cultured HTFs expressed TGF-β and CTGF. The mRNA expression of TGF-β and CTGF significantly decreased compared with the control group(t=20.042, 14.995, P<0.05). the protein expression of TGF-β and CTGF also decreased significantly compared with the control group(t=46.058、12.452, P<0.05)

CONCLUSION: The cultured HTFs can express TGF-β and CTGF in mRNA and proteins' level. LiCl can reduce the expression of TGF-β and CTGF both in gene and proteins' level. LiCl has the potential to modulate wound healing for glaucoma filtration surgery.]]> 2017/8/22 10:31:08 Su-Su Lu, Shan-Shan Liu, Xiao-Jun Fan, Xiao-Xiang Sun, Jiang-Hua Bian and Ji-Bing Wang 123true METHODS: The oxidative injury model of ARPE-19 cell was induced by exposure to various concentrations of hydroquinone(HQ)to determine the optimal concentration. Intestinal absorption solutions of BSYXMM Formula were prepared. Effect of intestinal absorption solutions of BSYXMM Formula on the cell viability was detected by CCK-8 assay, and the percentage of apoptotic cells was measured by TUNEL assay. The levels of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)in ARPE-19 cells were detected by means of chemical colorimetry.

RESULTS: It was found that ARPE-19 cell viability significantly decreased when the concentration of HQ was higher than 90μmol/L. Compared with the model group, 1% and 2% intestinal absorption solutions in the pre-treatment group could significantly alleviate HQ-induced injury(P<0.01)and 0.5% and 5% intestinal absorption solutions in the pre-treatment group could alleviate the injury in certain degree(P<0.05). While in the treatment group 1% and 2% intestinal absorption solutions could alleviate the injury to some extent(P<0.05). TUNEL results showed that the apoptosis rate decreased significantly in the pre-treatment group(P<0.01)and to some extent in the treatment group(P<0.05)compared with the model group. It was shown that both levels of SOD and GSH-Px in pre-treatment group and treatment group were markedly higher than that of model group(P<0.05), and pre-treatment group had more significant effect(P<0.01,P<0.05).

CONCLUSION: BSYXMM Formula could protect against HQ-induced oxidative stress injury in ARPE-19 cells, which may be related with the increasing of antioxidant enzyme in the cells.]]> 2017/7/24 10:38:38 Qiang Chen, Na An, Zeng-Yuan Zhuang and Li-Na Liang 122true METHODS: Inflammatory environments of human corneal stromal cells were simulated by using 1.5ng/mL IL-1β. The cells were then treated with or without different concentrations of estrogen(0, 1×10^-4, 1×10^-6, 1×10^-8, 1×10^-10mol/L estradiol)in vitro. Cell viability was evaluated by MTT. Expression levels of MMP-2, TIMP-2 and TGF-β1 proteins were measured by enzyme-linked immunosorbent assay(ELISA).

RESULTS:Estrogen did not affect the viability of human corneal stromal cells. Compared with the control group, expression levels of MMP-2 and TGF- β1 proteins in E2 treatment group significantly decreased after being treated with estrogen, while the expression level of TIMP-2 significantly increased.

CONCLUSION: Estrogen could, to some extent, down-regulate the expression of MMP-2 and TGF- β1 and up-regulate the expression of TIMP-2, which might contribute to protecting human cornea.]]> 2017/6/26 10:38:49 Yan Tian, Hong-Bo Yin and Ying-Ping Deng 121true METHODS:Totally 60 SD rats were randomly divided into 3 groups(20 in each group), 0.5mg/kg(in low dose group), 1.5mg/kg(in high dose group)tunicamycin were injected into vitreous cavity and saline(9g/L NaCl)were injected in the same dose as a control group. Changes of retinas were observed by OCT on the 1,7 and 14d after treatment of tunicamycin. Then the rats were sacrificed, retinas were taken out and embedded by the paraffin, tissue sections and the HE staining were performed.

RESULTS:OCT results suggested that tunicamycin played damage effects on retinal morphology and structure which appeared a time- and dose- dependent. Fundus photography results suggested that 2wk after tunicamycin treatments, with the gradually changing of tunicamycin concentration, peripheral retinal and macular region became pale color gradually, edema occurred in optic disk, retinal vessels appeared thinner in the high dose group, optic nerve came out atrophy. Fluorescein angiography confirmed that tunicamycin injection in vitreous cavity 2wk later, retinal vessels injury occurred, resulted in leaking of intravascular contrast agent from peripheral to the central part of the retinas. Electrophysiological data showed that retinal electrogram occurred disorder induced by tunicamycin, such as the amplitude of a wave, b wave decreased gradually, even closed to zero, which was very different from control significantly(P<0.05). HE staining of paraffin sections showed that retina injuries induced by tunicamycin were in dose - time dependent, which was consistent with the results of OCT.

CONCLUSION: Clinical retinal diseases could be simulated by retinal damage animal model induced by tunicamycin treatment. OCT detection offered real-time images of the retinal cross-section, which provided a helpful non-invasive method for detecting and evaluating the retinal damages.]]> 2017/6/26 10:38:49 Bo-Yi Zhang, Ya-Qiong Zhang and Hui-Xin Che 120true METHODS: Twenty-six pigmentation rabbits were randomly divided into normal control group(n=2), conventional photocoagulation group(n=6)and subliminal micropulse laser photocoagulation group(n=18). The conventional photocoagulation group was treated with 577nm laser photocoagulation, subcutaneous micro-pulsed laser photocoagulation at a working loading rate of 9%, 12% and 15%, respectively. Eighteen rabbits were again divided into three subgroups according different powers of subthreshold working loading rate of 9%(n=6), 12%(n=6)and 15%(n=6)that undertook, respectively. The expression of MMP-9 on the retina of rabbit eyes was detected by immunohistochemistry.

RESULTS: In the conventional photocoagulation group, the expression of MMP-9 in the RPE layer and the visual cell layer was strongly positive, which was significantly higher than that in the sub-micro pulse group(P<0.05). Less MMP-9 positive expression of RPE layer and visual cell layer in the working loading rate of 9% subgroup, and more MMP-9 positive expression of RPE layer and visual cell layer in the working loading rate of 12% subgroup; little expression was also noted in the nucleus of RPE and visual cell layer. Moderate MMP-9 positive expression of RPE layer and visual cell layer was observed in the working loading rate of 15% subgroup. There were no significant differences between the three subgroups(P>0.05).

CONCLUSION: The 577nm subliminal micro-pulsed photocoagulation has high selectivity to retinal pigment epithelium at working load rate of 9%, 12% and 15%, and no damage to retinal nerve fiber layer, which is safer than conventional 577nm laser photocoagulation.]]> 2017/6/26 10:38:49 Chun-Yan Wang, Hui-Li Li, Xiao-Dan Li, Hai Yu and Yan Fu 119true METHODS: Experimental CNV models were induced by semiconductor laser in 30 male Brown Norway(BN)rats and randomly divided into 3 groups with 10 rats in each group. At 21d after photocoagulation, the single administration group were given intravitreous injection with Ad-Es 0.01mL; the repeated administration group were given intravitreous injection with Ad-Es 0.01mL and a repeated injection 7d later; the saline control group were given intravitreous injection with saline 0.01mL. At 7d after final administration, the leakage of fundus fluorescein angiography(FFA)was observed. Various CNV areas were measured by using laser confocal microscopy of choroidal flatmount method. Pathology and ultrastructure were observed with light microscopy, the expressions of CD105 were measured by immunohistochemistry.

RESULTS: The leakage of CNV of the administration group abviously decreased as compared with those in the saline group, the leakage of repeated administration group decreased compared with that of single administration group(P<0.05). Laser confocal microscope quantitative CNV analysis showed that CNV area of the administration group was significantly smaller than that of control group, the area of repeated administration group was smaller than that of single administration group(P<0.05). Under the light microscope, the vascular endothelial cell number and quantity of the administration group were significantly lower than that of the control group, the cell number of repeated administration group was lower than that of single administration group. CD105 expression of the administration group was significantly weaker than that in the saline group; the expression of repeated administration group was weaker than that of single administration group.

CONCLUSION: Ad-Es can effectively inhibit semiconductor laser induced CNV in BN rats, and the inhibition effect of repeated administration group is better than that of single administration group. It may be a useful new method in the treatment of CNV.]]> 2017/5/24 16:26:29 Li-Juan Chen,He-Xiang Song and Lin Miao 118true in vitro.

METHODS: RF/6A cells in good conditions were administrated with DMSO(the control group), apelin-13 at 0.1μmol/L(low dose group)or apelin-13 at 1μmol/L(high dose group). Cell proliferation, migration and capillary-like tube formation were detected by using the MTT assay, scratch assay and matrigel assay, respectively, at 24h after plating the cells.

RESULTS: Cell proliferation was promoted in both low and high dose apelin-13 groups compared to the control cells(P<0.05); the cell migration distance of both apelin-13 groups was significantly greater than that of the control group(P<0.05); and the number of capillary-like tube structures of both apelin-13 groups was significantly larger than that of the control cells(P<0.05). In addition, cell proliferation, migration and tube formation increased as the concentration of apelin-13 increased.

CONCLUSION: Apelin-13 could obviously promote the angiogenesis capacity of RF/6A cells, suggesting that apelin-13 was an important pro-angiogenic factor in retinal endothelial cells.]]> 2017/5/24 16:26:29 Kun-Peng Xie,Ping Liu,Xin Wang and Jun-Hui Du 117true METHODS: A total of 60 SD rats were randomly divided into the blank control group, model group and PEDF group, with 20 rats in each group. Except the blank control group, the optic nerve crush injury rat models were established in the other groups, and left eyeballs were taken as samples. After successfully modeling, the model group were treated with intravitreal injection of 5μL of balanced salt solution while PEDF group were treated with intravitreal injection of 5μL of PEDF(0.2μg/μL). Two weeks later, the retinal tissues were collected, and changes of shape were observed under microscope after HE staining. The changes of NO level were measured by colorimetry assay, the expression of caspase-3 mRNA and caspase-3 protein was detected by reverse transcription-polymerase chain reaction(RT-PCR)and Western-blot.

RESULTS: HE staining showed that retinal tissues of the blank control group arranged neatly and clearly. Retinal ganglion cells(RGCs)arranged in a monolayer, and cells were oval, uniform in size and distribution, the cell nuclei were clear, closely arranged, with clear boundaries. The retinal tissues of the model group were sparse in shape, RGCs showed vacuolar changes, the overall number of cells was reduced, and cell nuclei of residual RGCs showed pyknosis and uneven staining. RGCs in PEDF group were with slightly edema and arranged closely, and the degree of injury was significantly milder than that in the model group. Levels of Caspase-3 mRNA and protein and NO levels in the three groups showed the model group > PEDF group > blank control group(all P < 0.05).

CONCLUSION: The application of PEDF can down regulate the expression of Caspase-3 and NO in rates with optic nerve injury and reduce RGCs injury.]]> 2017/5/24 16:26:30 Xiao-Xiao Yan,Hai-Bo Jia,Xiao-Ling Yin,Cui Cui,Wei-Xing Pu,Nan Huo and Jun-Bo Zhao 116true METHODS: Twenty-four male Brown Norway(BN)rats were randomly divided into the control group(6 rats)and the experimental group(18 rats). The CNV were induced by 659nm krypton laser. And the power was 360mW, exposure time was 0.05s, spot diameter was 50μm. The formation rate of CNV and the average optical density(AOD)of leakage flare were observed by fluorescein fundus angiography(FFA)and indocyanine green angiography(ICGA)on the seventh day, the fourteenth day and the twenty-first day after laser photocoagulation. The histopathology changes of CNV and the AOD of CD105 were observed by eyeball exemplar.

RESULTS: The CNV appeared on the seventh day after laser photocoagulation, and reached the peak on the fourteenth day and twenty-first day after laser photocoagulation. The formation rate of CNV were 77.08%, 85.42%, 89.58%, at 7, 14 and 21d after laser photocoagulation. From 7 to 21d after laser photocoagulation, the AOD of leakage flare increased gradually(P<0.05). There had significant differences between 7 days' outcome and 14 days' outcome(P<0.05), and had no significant differences between 14 days' outcome and 21 days' outcome(P>0.05). From 7 to 21d after laser photocoagulation, the expression of CD105 increased gradually, and then decreased gradually, and from 14 to 21d, there had significant differences between 7 days' outcome and 14 days' outcome of AOD(P<0.05), and had no significant differences between 14 days' outcome and 21 days' outcome(P>0.05).

CONCLUSION: Immunohistochemical outcomes of CD105 are highly consistent with fundus angiography outcomes of CNV changing regularity.]]> 2017/4/25 17:20:01 Fang-Fang Tao,Ze-Feng Kang,Jian Liu,Wen-Li Chu and Zhi-Hao Zhou 115true METHODS: Totally 28 standard rabbits were selected with right eyes operated for vitrectomy, and then, they were randomized into three groups: Group A 12 rabbis, Group B 12 rabbits, Group C 4 rabbits. Group A: vitreous body was cut-down and filled with silicon oil; Group B: vitreous body was cut-down and filled with heavy silicon oil; Group C: vitreous body was cut-down and filled with BSS. Taken into account were the experimental animals' different periods thickness of the retinal, intraocular pressure and ERG b-wave amplitude for statistical analysis.

RESULTS: Comparison between any two means of Group A, B or C surgery eyes' preoperative and postoperative at different time points: measured IOP showed no significant difference(P> 0.05); the retinal thickness mean value measured by OCT had statistically significant(P<0.05)at the postoperative 24wk; there was a conspicuous statistically significant(P<0.01)of ERG'sb-wave amplitude at the postoperative 24wk.

CONCLUSION: As the stuffing of vitreous cavity, the silicone oil or heavy silicone oil has no obvious difference to the influence of intraocular pressure for medium to longer term. But heavy silicone oil has more serious negative impact of retinal visual information transmission function, more significantly reduce of retinal thickness than ordinary silicone oil in the longer term.]]> 2017/4/25 17:20:01 Wen-Hua Duan and Yun-Hai Jiang 114true METHODS: Thirty-eight cases(38 eyes)of CCh were recruited as the CCh group and 36 cases(36 eyes)without CCh were included as the normal control group and the conjunctival tissue and tear were collected, respectively. Mucins(MUC2, MUC4, MUC5AC, MUC16)expression in conjunctival tissue were detected by immunohistochemical staining; Mucins(MUC2, MUC4, MUC5AC, MUC16)OD value in tear were determined by ELISA, the differences of which were compared between the CCh and normal control group.

RESULTS: No difference in the expression of MUC2 and MUC4 in conjunctival epithelia was observed between the two groups(P=0.315, P=0.156). There was significant difference in the expression of MUC5AC and MUC16 in conjunctival epithelia between the two groups(P=0.016, P<0.01, respectively). No significant difference in the OD value of MUC2 in tear was found between the two groups(P=0.651). The OD value of MUC4 and MUC5AC in tear were significantly lower in CCh group that in the normal control group(P<0.01, P<0.01, respectively), so was the OD value of MUC16(P=0.022).

CONCLUSION: MUC5AC and MUC16 both decreased in the conjunctiva and tear of eyes with CCh. The further study may reveal the pathogenesis of conjunctivochalasis.]]> 2017/4/25 17:20:01 Min-Hong Xiang,Xing-Xing Chen,Huan-Ming Zhou,Yuan-Ling Jia,Qing-Song Li,Xing-Ru Zhang and Xia Sheng 113true METHODS: The freshly excised corneas,whole corneas or de-epithelialized corneas, were immediately mounted in perfusion cells. Dexamethasone containing free-borneol or borneol was added into the deliver chamber respectively in each group. At different time point after addition of solution, 50μL of the receptor solution was removed. The concentration of dexamethasone was determined and the apparent permeability coefficient(Papp)was calculated. Hydration level of cornea was measured and irritation test was observed.

RESULTS: The Papp for dexamethasone in whole cornea increased by 2.40 fold in borneol group(P<0.05). The Papp for dexamethasone in de-epithelialized cornea did not increased apparently(P>0.05). Synthetic borneol did not increase hydration level and irritation of cornea(P>0.05).

CONCLUSION: Synthetic borneol can improve the corneal permeation of lipophilic drug dexamethasone and do not damage corneal tissue.]]> 2017/3/27 10:17:04 Hong-Bin Yang,Jie Jiang,Xiao-Yu Zhang,Yan-Bin Xun,Hao Cui and Qiang Guo 112true METHODS: The expressions of miR-146 and IL-6 were examined in RPE in mice aged from 2mo to 24mo by qRT-PCR. Then, the expressions of miR-146 and IL-6 in RPE of wet ARMD patient were examined also. Finally, the effect of overexpression of miR-146a in APRE-19 cell line on expression of IL-6 was checked.

RESULTS: MiR-146 was positive correlated with age, and the expression of IL-6 had no change in aging RPE. However, the expression of miR-146 decreased and IL-6 increased in RPE of ARMD. In cultured APRE-19 cells, overexpression of miR-146 inhibited the expression of IL-6 induced by TNF-α.

CONCLUSION: Our results suggest that there is a biological correlation among the development of ARMD, expression of IL-6 and miR-146 in aging RPE. It also suggests that, on the one hand, regulation between IL-6 and miR-146 may be important for the clinical treatment, on the other hand, both IL-6 and miR-146 can be potential molecular markers for diagnosing ARMD.]]> 2017/3/27 10:17:04 Yi Hao,Xiao Sun,Yu-Qiu Liu,Yun Zhao and Yu Wang 111true METHODS: Ninety-six 7-day-old SD rats were randomly divided into 6 groups: Group Z:Normal group; Group O: OIR; Group A: OIR + vehicle control group; Group B: OIR+5μg celecoxib group; group C: OIR + 20μg celecoxib group; group D: OIR + 80μg celecoxib group. In addition to Z group in the normal environment, the other groups were established the OIR model. The neonatal rats were given intravitreal injections of dimethyl sulfoxide(DMSO)and the corresponding doses of celecoxib on the 12th day after birth. The rats were sacrificed on day 17 after birth. HE staining were employed to count the vascular endothelial cells which were breakthrough within the internal limiting membrane of retina. Immunohistochemistry staining were utilized to probe the expression of VEGF protein.

RESULTS: HE staining showed that, the number of the endothelial cells in the retina was 0.44±0.18, 30.60±5.36, 28.05±4.68, 19.58±4.58, 10.13±1.93, 7.58±2.68 in Group Z, O, A, B, C and D. In addition to Group O and Group A, there were significant differences between the two groups(P<0.05). After treatment with celecoxib, breakthrough of the internal limiting membrane of the vascular endothelial cell nucleus was significantly reduced, and positively correlated with the dose. Immunohistochemical results showed, the expression of VEGF protein in Group Z was negative, the expression rate was 10%,the positive expression of VEGF protein in Group A was higher than that in Group B, C and D, and the positive rate was 86%, which was higher than that of Group B, C and D, as 68%,42%, 30%.

CONCLUSION: Celecoxib can inhibit the OIR model of rat retinal angiogenesis, and the effect of suppressing a positive correlation with the dose,its action may inhibit VEGF expression.]]> 2017/3/27 10:17:04 Peng Li,Bing Ren,Xiao-Wei Gao,Yan Cai,Yan Ju and Na Yi 110true METHODS: Totally 22 healthy rabbits were randomly divided into 2 groups, which were experimental group and normal control group. Both eyes of rabbits in experimental group were chosen to establish corneal neovascularization model by alkali burn. The morphologic change of corneal was observed with slit lamp microscope and the area of CNV was calculated every day. After alkali burn, the right eye of the experimental group was accepted EPO siRNA injection under the conjunctiva, and the left eye was assigned to be experimental control group. The corneal with CNV was collected for immunohistochemistry at 3d, 7d, 14d, 21d after alkali burn, and the expression of EPO was measured.

RESULTS: CNV began growing at the 3d after alkali burn in experimental group, and it was vigorous growing at 7d-14d period. The result of immunohistochemistry shows that the expression of EPO increased after the operation. Compared with experimental group, the rabbits who were treated by EPO siRNA was found with less neovascularization on their corneal, and the expression of EPO decreased. There were statistical significance between the two group at different time(P<0.05).

CONCLUSION: EPO is likely to play an important role on CNV growth, and EPO siRNA can inhibit the growth of CNV by restraining the expression of EPO.]]> 2017/2/27 10:55:37 Yu-Shun Xue,Li Qin,Le Yang,Tao Zhu and Rui Shi 109true METHODS: The experiment was divided into AMD patients, cataract patients and normal people group. AT1R was the target gene of miR-410 by bioinformatics, and the normal RPE cells were cultured in the simulated microenvironment of AMD and cataracts and the expression of miR-410 was detected. Then miR-410 mimics was transfected into cells, and the expression of mRNA and protein of AT1R were detected by Q-PCR and Western blot respectively. The relationship between miR-410 and AT1R was confirmed by the dual luciferase reporter assay.

RESULTS: The miR-410 expression of in RPE cells with AMD was significantly reduced(P=0.0006, 0.0008)compared with cataract and normal controls. The miR-410 can regulate the function of AT1R by dual luciferase reporter gene experiment and the inhibition rate was about 40%. In addition, miR-410 inhibition rate was about 40%-50% to AT1R mRNA and protein expression by cell experiment.

CONCLUSION: AT1R was a target gene of miR-410 in cell experiments, and it is demonstrated that increasing the expression of miR-410 in RPE cells with AMD can suppress the expression of AT1R.]]> 2017/2/27 10:55:37 Xia Li 108true METHODS: Totally 60 SD rats aged 7d were randomly divided into 4 groups: normal group, model group, intervention group and NS control group. Normal group was raised in a normal air feeding; model group at 75% high oxygen for 5d to establish the model of oxygen induced retinopathy; intervention group was given anti p16 methylation drug 5-aza-CdR(0.25 mg/kg)intraperitoneal injection; NS control group was given the same volume NS intraperitoneal injection. The eyes were taken from each group and the left eyes were removed for observation of retinal neovascularization by HE staining, and immunohistochemistry and immunofluorescence were taken for observations of p16 protein expression. Right retina had been performed real time-PCR to analysis p16mRNA expression.

RESULTS: The normal group were not found retinal neovascularization breaking through internal limiting mebrane. In model group and NS control group, the retinal tissue was obviously thickened, and a large number of new blood vessels were found. In the intervention group, a small amount of new blood vessels were found in the retina. The expression of p16 was low in the model group, the positive cell number was 19.52±2.67, and the number of the positive cells was 36.38±3.16 in the intervention group, the difference was statistically significant(P<0.001). Real time-PCR showed decreased expression of p16 mRNA in the model group(2^-△△ct=0.14±0.01), the expression of p16 mRNA in the intervention group rats retina was significantly higher than that of NS control group rat retina, there was significant difference between two groups(2^-△△ct=0.68±0.08, P<0.001).

CONCLUSION: The abnormal expression of P16 may be closely related to the proliferation of retinal neovascularization. Inhibition of p16 methylation can decrease the proliferation of retinal neovascularization.]]> 2017/1/20 11:21:30 You Li, Dong-Bo Pang and Xiao-Long Chen 107true METHODS: Resected specimens were collected from patients suffering from B-cell non-Hodgkin lymphoma(NHL)(n=40)and lymphadenosis(n=20)of ocular adnexal in Affiliated Hospital of Qingdao University from June 1995 to December 2015. Lymphadenosis of ocular adnexal was acted as the control group. PCR and immunohistochemistry were employed to examine the MTDH and β-catenin mRNA and protein expression respectively. The relationship between the MTDH and β-catenin protein expression level and the clinical pathological characteristics were analyzed.

RESULTS:The expression of mRNA and protein of MTDH and β-catenin in ocular adnexal lymphoma lesions were higher than that in control group(P<0.05). The expression of MTDH and β-catenin proteins were related to pathologic type of tumors, but not related to age, gender or pathogenic site. With the increase of pathologic grade, MTDH and β-catenin labeling frequency increased gradually. And there was a positive correlation between MTDH and β-catenin(r=0.389, P=0.036).

CONCLUSION: Over expression of MTDH and β-catenin may play a significant role in the ocular adnexal lymphoma. The expression of MTDH and β-catenin has a positive relationship.]]> 2017/1/20 11:21:30 Yan Gao, Hong Lin, Dan Qi, Jing Lin, Li-Ting Hu and Gui-Qiu Zhao 106true METHODS: Activity of GPx, MDA level in lens and selenium content in the eyeballs of different ages rats were determined. Besides, lanthanum hydroxide \〖La(OH)₃\〗 tracer method was used to detect development status of blood-retina barrier at different ages.

RESULTS: The result showed that the enzyme activity of GPx was highest in young rats before open eyes, but then decreased gradually with age. Distribution of La(OH)₃ in retinal pigment epithelial layer of 20-day-old rats was significantly less than 11-day-old rats. Injecting sodium selenite to 9-day-old rats, lanthanum hydroxide increased obviously and extended to the inner layers of the retina after 48h, and the retinal pigment epithelial layer was damaged seriously; while injecting sodium selenite to 18-day-old rats with the same dose, number of lanthanum hydroxide decreased significantly and did not extend to the inner layer after 48h.Before opening eyes, the content of MDA in the lens of rats was the highest, and decreased significantly after opening eyes. The Se group was 5 times as that of the control group. Besides, in these groups of rats, selenium content in the eyeballs and MDA level in the lens were in agreement with the change of La(OH)₃ distribution.

CONCLUSION: These results indicated that antioxidant capacity in the eyelid unopened rats is not the main reason for selenite induced cataract formation. The real reason is that blood-retina barrier development is not mature in the eyelid unopened rats.]]> 2017/11/20 16:09:09 Ping Lu 105true METHODS: Totally 42 healthy rabbits(84 eyes)provided by the Animal Experimental Center of our hospital were selected, including 18 female rabbits, 24 male rabbits, average weight 2.24±0.31kg, and they were randomly divided into 6 groups, 7 rats in each group(14 eyes). We observed the change of intraocular pressure after laser iridectomy surgery at 20min, 2, 6, 18, 24h and the nitric oxide(NO), malondialdehyde(MDA), superoxide dismutase(SOD), 6-keto-prostaglandin(6-keto-PGF1α)and nitric oxide synthase(NOS)content in aqueous.

RESULTS: There was no significant difference in intraocular pressure, NO, NOS, SOD, MAD and 6-keto- PGF1α before operation(P>0.05). The intraocular pressure increased after operation, and the difference was statistically significant(P<0.05)at 20min, 2 and 6h after operation, and decreased at 18h after operation, 24h after operation(P>0.05). The levels of NO, NOS and SOD in the aqueous humor of the two groups decreased 20min, 2 and 6h after the operation(P<0.05), while increased after 6h, increased more at 18 and 24h.The difference with control group was no more significant(P>0.05). The levels of MDA and 6-keto-prostaglandin in the aqueous humor increased after the operation, and the difference was statistically significant at 20min, 2 and 6h after operation(P<0.05), while decreased at 18 and 24h and the difference with control group was not significant(P>0.05).

CONCLUSION: The increase of transient intraocular pressure after laser iridectomy may relate to the increase of malondialdehyde, 6-keto-prostaglandin content and the decrease of superoxide dismutase and nitric oxide in the aqueous humor after operation.]]> 2017/11/20 16:09:09 Zhi-Juan Pei 104true METHODS: Y79 cells were cultured in RPMI1640. The cultured cells were stimulated with 0.25μmol/L of oxaliplatin. The expression of Mcl-1 protein was detected by Western blot after 6, 16 and 24h respectively. Cells in logarithmic phase were collected and used for single-cell suspension. Then they were transfected with empty plasmid, Mcl-1-homo-991, Mcl-1-homo-1114 and Mcl-1-homo-1235. After 6h, fluorescence microscope was used to observe the transfection efficiency and the optimal one was selected. The cells were divided into Group A and transfected with empty plasmids. The cells transfected with Mcl-1 were divided into Group B and Group C. Group A and Group C were treated with 0.25μmol/L oxaliplatin for stimulating induction, and the apoptotic rate was compared after 24h.

RESULTS: The expression of Mcl-1 in Y79 stimulated by oxaliplatin was the most after 24h of culture. Mcl-1-homo-991 significantly inhibited the expression of Mcl-1 in Y79 after transfection. There was no significant difference in the apoptosis rate in Group A(11.1%±1.2%)and in the control group(6.1%±0.6%)(P>0.05). The apoptotic rate of Group C(49.2%±2.7%)was significantly higher than that of Group B(20.8%±1.9%). At the same time, the apoptotic rates of these two groups were significantly higher than those of Group A and control group, the difference was statistically significant(P<0.05).

CONCLUSION:Downregulation of Mcl-1 by siRNA can reduce the drug resistance of Y79, thereby enhancing the apoptosis of Y79, and reducing the survival rate of Y79.]]> 2017/11/20 16:09:09 Lu Zhou and Na Li 103true 2O₂ induced oxidative stress damage in human lens epithelium cells(HLEC).

METHODS: HLEC were cultured and divided into negative control group: cultured in normal cultivation; oxidative damage group: treated with 100μmol/L H₂O₂ for 12h; Salidroside low concentration group: 10μmol/L salidroside treated for 24h and H₂O₂ treated for 12h; Salidroside high concentration group: 100μmol/L salidroside treated for 24h and H₂O₂ treated for 12h. MTT method was applied to observe the effect of salidroside on HLEC survival rate. Morphological change of each group were observed and recorded under inverted microscope. DCFH-DA fluorescent probe was applied to detect intracellular ROS changes; content of malondialdehyde(MDA), superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)in supernatants were detected by pectrophotometer.

RESULTS: Salidroside obviously inhibited H₂O₂-induced HLEC vitality decline, inhibited ROS generation in cells, causing SOD, GSH-Px levels increased and MDA levels decreased.

CONCLUSION: Salidroside inhibited H₂O₂ induced HLEC injury by decreasing the intracellular MDA content levels and increasing SOD, GSH-Px content levels, which conclude that salidroside may have a certain role in the treatment of HLEC damage.]]> 2017/9/18 10:53:56 Li-Ting Liu, Wen-Tao Zheng, Ping Liu and Li-Juan Zhang 102true in vitro by observing its effects on proliferation, migration and capillary-like tube formation of murine retinal vascular endothelial cells(RVECs).

METHODS: Well cultured RVECs were divided to different groups which were treated with 0, 5, 10 μmol/L LCN-2 for 48h, respectively. Cell proliferation, migration and tube formation were detected by using the EDU assay, transwell assay and matrigel assay, respectively.

RESULTS: The cell proliferation rate was promoted in both low and high dose of LCN-2 groups compared to the control cells(P<0.05). The number of migrated cells in both LCN-2 groups was significantly larger than that of the control group(P<0.05). The number of capillary-like tube structures of both LCN-2 groups was significantly larger than that of the control cells(P<0.05). In addition, cell proliferation, migration and tube formation were all increased with the increase of LCN-2 concentration.

CONCLUSION: LCN-2 could obviously promote the angiogenesis capacity of RVECs, suggesting that LCN-2 is an important pro-angiogenic factor in retinal angiogenesis.]]> 2018/8/17 17:07:07 Guo-Min Yao, Rong Li and Jin Tian 101true METHODS: Cultured cell line ARPE-19 was divided into control group, H₂O₂ treating group, Mcc950 treating group and Mcc950+H₂O₂ treating group. Different concentrations of H₂O₂ and Mcc950 were used to treat the cells. Cell activity was detected by using CKK8 and proper experimental concentration of H₂O₂ and Mcc950 were determined. After the treatment, the concentration of IL-1β were detected by using ELISA. The change of NLRP3 related proteins were detected by Western blot. And cell apoptosis was examined by TUNEL stain.

RESULTS: Cell ability was gradually decreased along with the increasing treating concentrations of H₂O₂. Cell ability showed statistical difference when the concentration of H₂O₂ arrived 400μmol/L. With the concentration of 0.1 and 1μmol/L, Mcc950 had no effect on cell ability. So we chose 400μmol/L H₂O₂ and 1μmol/L Mcc950 as the experimental concentrations. Compared with the normal control group, the cell viability in the H₂O₂ treating group was significantly reduced, the IL-1β in the supernatant was significantly increased, and the apoptosis rate was significantly increased, with statistically significant differences(P<0.05). In Mcc950+H₂O₂ treating group, cell viability was significantly increased, the IL-1β in the supernatant and the apoptosis rate were significantly decreased(P<0.05). By Western blot, after treated with 400μmol/L H₂O₂, the IL-1β, NLRP3, pro-caspase1 and caspase1 were obviously increased compared to control group. After treated with Mcc950, the NLRP3 and pro-caspase1 still were at high level, the expression of caspase1 was suppressed, which indicated that Mcc950 effectively inhibited the activation of NLRP3 inflammasome consequently disturbed the formation of caspase1.

CONCLUSION: Mcc950 can inhibits the function of NLRP3, leading to increasing of the cell ability and decreasing of the cell apoptosis.]]> 2018/8/17 17:07:07 Shu-Yan Li, Xiao-Feng Li, Xing-Zhao Xu and Qing-Huai Liu 100true METHODS: Cells were treated with 100nmol/L and 1 000nmol/L insulin for 48h respectively. Expression of protein and mRNA were detected by western blot and quantitative real-time polymerase chain reaction. Cellular proliferation and permeability were examined by methods of methylthiazolyl tetrazolium and horseradish peroxidase.

RESULTS: With treatment of insulin, protein and mRNA of syndecan-1 both increased obviously, and the effect of high level insulin was more significant. After treated with insulin, cellular proliferation and permeability both enhanced, and the effects of high level insulin were stronger.

CONCLUSION: Insulin can up-regulate syndecan-1 protein and mRNA in cultured human retinal microvascular endothelial cells, and increase cellular permeability and proliferation.]]> 2018/7/20 11:38:05 Yong-Liang Hu, Jing-Bo Wang, Li Zhou, Xiao-Xia Han and Ling-Hao Shi 99true METHODS:Well-grown ARPE-19 cells were collected for experimentation. The standard curve of CCK-8 was made to determine the proper cell density of ARPE-19 cells and the reaction time of CCK-8 reagent. The cells were divided into dark group, blue light group and white light group, which were irradiated for 3, 6 and 9h respectively. After 12h of light-repellent treatment, CCK-8 method was used to examine the effects of different light sources on the proliferation rate of ARPE-19 cells at different times.

RESULTS: The number of cells per well was selected by CCK-8 standard curve to be 20 000, and the corresponding absorbance value was measured after 4h of the action of CCK-8 solution. The CCK-8 test results showed that the cell proliferation rates of the three groups were significantly different(P<0.01). The cell proliferation rate of the blue light group was significantly different(P<0.001)at 3, 6 and 9h, and the cell proliferation rate decreased gradually with the extension of the illumination time. The cell proliferation rate of the white light group was significantly different(P<0.05)at 3, 6 and 9h; there was a statistically significant difference in the rate of cell proliferation at 3h and 6h in white light(P<0.05), however, there was no significant difference in the rate of cell proliferation at 9h illumination compared with 3h and 6h illumination respectively(P=0.253, 0.120). The proliferation rate of cells under white light for 3-6h showed a downward trend, while that of cells under light for 6-9h showed an upward trend. At the same illumination time, the proliferation rate of the cells in the blue and white groups was lower than that in the dark group, and the cell proliferation rate in the blue group was lower than that in the white group. The difference was statistically significant(P<0.05).

CONCLUSION: The proliferation of ARPE-19 cells was inhibited by light irradiation. The proliferation rate of cells in blue light group was significantly lower than that of white light group. With the increase of light time, the cell proliferation rate decreased.]]> 2018/7/20 11:38:05 Sheng Chen, Ke Liu, Shan Qin and Shen-Wen Liu 98true METHODS: DM was induced in SD rats by the administration of streptozotocin(STZ). DM rats were divided into two groups: COX-2 treated group(CTG, n=15), and diabetic group(DG, n=15). Normal control group(NCG)consisted of 15 rats was given. Three months' treatment was given after DM development. Sample of retina were made at the end of 3mo. Expression of Cx43 in retina were investigated by immunohistochemistry. Expression of Cx43 mRNA in retina were analyzed by RT-PCR.

RESULTS: Cx43 was expressed in the ganglion cell layer, the nerve fiber layer, the inner plexiform layer, and the pigment epithelium layer of retina with a different degree. Computer image analysis showed that celecoxib had elevated the expression of Cx43 in experimental diabetic rats, gray values were: 0.233±0.025, 0.124±0.014, 0.197±0.021; multiple comparisons found statistically significant difference(P<0.05). Celecoxib had raised the expression of Cx43 mRNA in diabetic rats with real time quantitative PCR. Relative value was 0.635±0.084, 0.110±0.061, 0.367±0.074, multiple comparisons found statistically significant difference(P<0.05).

CONCLUSION: Expression of Cx43 decreases in retina of SD rats with DM. Celecoxib can relieve the decrease of Cx43 expression in retina of them.]]> 2018/7/20 11:38:05 Gui-Ping Pan, Zhen Tian, Yong Zhang, Jun Yuan and Ya-Qin Wang 97true in vitro and the mRNA express of apoptotic inhibitor B-cell lymphoma(Bcl-2), Bcl2-Associated X(Bax)and apoptotic actuators Caspase-3 in different groups.

METHODS: Cultured human RPE cells were divided into different group in random. According to the time of exposing, the four groups were 3h group, 6h group, 12h group and no light-exposed group. Cells of different groups were exposed to the opening mobile phone that was kept high-light brightness and playing on the colorful picture on silent mode. The morphological and functional change of RPE was quantified after exposed by HE staining, TUNEL staining, MTT, and use the technology of polymerase chain reaction(PCR)to detect the mRNA express of apoptotic inhibitor Bcl-2, Bax and apoptotic actuators Caspase-3 in different groups.

RESULTS: The different was not statistically significant(P>0.05)in the way of HE staining, TUNEL staining and MTT. The expression of Bcl-2, Bax and Capase-3 intracellular was no difference statistically significant in 3h, 6h group to no exposure group on the way of PCR(P>0.05). But as the time of exposed increased to 12h, the expression of apoptotic inhibitor Bcl-2 decreased, at the same time the expression of apoptotic actuators Bax and Caspase-3 increased, and the different was statistically significant(P<0.05).

CONCLUSION: The human RPE would be injured under the constantly high-light brightness of mobile phone screen as the man-made light source.]]> 2018/6/27 10:52:29 Jin Zhou, Jiao Xie, Jie Lei, Xiang Ji and Qing Yang 96true METHODS: Totally 25 healthy male Japanese white rabbits were randomly divided into 5 groups: control group, model group, western medicine group, high dose of Qingxuan decoction group, low dose of Qingxuan decoction group. The blank control group did not do any treatment. The improved dry eye model of rabbit was prepared by the improved method of glandular burning of the eyelid plate. The high and low dose group were given daily 27.2mg/kg, 6.8mg/kg Qingxuan decoction by gavage. The model group was intragastric with the same amount of normal saline every day. The western medicine group with tobramycin and dexamethasone ophthalmic ointment 1 drops, once a day. The treatment were administered continuously for 28d. At 14d before the experiment, 7d before the experiment, 7d after the model, and 14d after the model, all the rabbits were tested by Schimer Ⅰ test(SⅠt)and break-up time(BUT). On the 15d after modeling, the animals were sacrificed by excessive anaesthesia. Rabbit ocular surface tissue sections were prepared. Hematoxylin-eosin staining method was used to observe the corneal morphological changes in each group. The concentrations of TNF-α, IL-1 and IL-6 in the ocular surface of rabbits were detected by ELISA.

RESULTS:(1)BUT, SⅠt: 7d after the model had been prepared, BUT and SⅠt of the model group and the western medicine group, high dose and low dose of Qingxuan decoction group was improved(P<0.05); Those of western medicine group, high dose and low dose of Qingxuan Decoction group compared with the model group, were significantly different(P<0.05).(2)TNF-α, IL-1, IL-6: The ELISA assay showed that TNF-α and IL-1, IL-6 concentration in the model group rabbits was significantly higher than those of the control group, TNF-α and IL-1, IL-6 concentration in western medicine group and high dose group of rabbits was significantly lower than those in the model group, the differences were statistically significant(P<0.05), and in high dose group the effect was better than that of Western medicine group.(3)Histopathological examination: on the 14d after the model, the corneal epithelium in the blank control group was stratified well. The cells in the base were columnar, near the surface, the cornea epithelium showed a squamous change. Conjunctiva showed complete epithelial layer and subconjunctival tissue layer, and goblet cells arranged closely. The number of corneal epithelial cells in model group was reduced or even stripped, and the matrix layer was disorder; Irregular loss of conjunctival epithelial cell layer and a large decrease in goblet cells. The corneal morphology of the rabbits in the western medicine group and the high dose group was close to the normal group, and the number of conjunctival goblet cells was not significantly different from that in the blank control group.

CONCLUSION: The expression of Qingxuan decoction can inhibit the inflammatory reaction through down-regulation of TNF-α and IL-1, IL-6 and in evaporative dry eye rabbit cornea and conjunctiva, so as to improve the ocular symptoms, increase tear secretion, prolong the time of BUT.]]> 2018/6/27 10:52:29 Ke-Xin Yu, Jing Yao, Jia-Di Wang and Cong-Hong Cao 95true METHODS: Totally 30 healthy guinea pigs were divided into 6 groups: Group I-II were fixed in 4% paraformaldehyde solution and 4% glutaraldehyde solution for 24h, respectively; Group III-V were fixed in Davidson solution for 3, 6 and 24h, respectively; and Group VI were fixed in Davidson solution for 3h and then transferred into 10% neutral formaldehyde solution for 48h. All groups were sectioned by routine section method and undergone PAS staining, and then observed by light microscope.

RESULTS: It was found that the group which was fixed by Davidson solution for 3h, remained the most complete structure for PAS staining(Group III). While the effect of fixation for the group which was transferred into 10% neutral formaldehyde solution for preserving for 48h after fixing in Davidson solution for 3h was also acceptable for PAS staining(Group VI). The retinal cells remained clear and in order for both groups which was mentioned above.

CONCLUSION: The best fixation condition for PAS staining for eyes of guinea pigs is fixation in Davidson solution for 3h among these fixation conditions, while it is also suitable to transfer the eyes into neutral formaldehyde solution after fixing in Davidson solution for 3h for preserving for long periods, which is not severely reduced the effect of fixation for PAS staining.]]> 2018/5/25 15:46:38 Hui-Xin Song, Wen-Jun Jiang and Hong-Sheng Bi 94true METHODS: Thirty SD(Sprague Dawley)rats were administrated Bangladesh via tail vein. Then 532nm laser(80mW, 100μm and 100ms)was performed on retinal vein secondary bifurcation of bitamporal optic disk for 50 spots. Electroretinogram(ERG), fundus fluorescein angiography(FFA), optical coherence tomography(OCT)and fundus(fluorescein)photograph were applied on 1, 3, 5, 7, 10, 14 and 21d after BRVO model constructed. Two rats were sacrificed, respectively, on 1, 5 and 21d after photocoagulation to carry on HE(Hematoxylin-Eosin stain)and VEGF-α(vascular endothelial growth factor-α)immumohistochemical staining.

RESULTS: There were three rats died, three rats with severe retinal detachment for excessive bleeding, one rat with retinal sunken, and one rat with cataract. FFA and fundus(fluorescein)photograph showed that the successful BRVO rat model was 73%(22/30). It was found that the near-end photocoagulation vein became coarse, far-end became diminution on 1d and the photocoagulation vein total recanalization was on 3-7d. ERG showed the amplification of b wave(dark-adaptation 3.0 response)decreased to 0.694±0.042 of control eyes and on 5-7d decreased to rock bottom about 0.487±0.064 of control eyes. Then it increased all the time to 0.708±0.0465 of control eyes on 21d. OCT and HE staining found that retinal ganglion cells and outer nuclear layer became edema on 1d in vivo and in vitro. It was observed that the thickness of retina on photocoagulation vein(0μm or 250μm)decreased from 5d and there were 3-4 layer cells in ONL on 21d. The expression of VEGF-α at injured site were significantly more than control eyes on 1d and there were no significant difference on 5d; But the expression of VEGF-α were slightly less than control eyes on 21d.

CONCLUSION: Photochemical method was a feasible method to establish BRVO rat model. The evolution and development of the BRVO model could partly mimic human BRVO phenomenon. At the same time, it should be improved to increase the successful model rate.]]> 2018/4/24 14:27:16 Pan Long, Wei-Ming Yan, Tao Chen, Jian-Cong Wang, Man-Hong Li, Jun-Hui Xue, Jing An and Zuo-Ming Zhang 93true METHODS: Totally 48 eyes from all the 24 New Zealand rabbits were inoculated with Staphylococcus aureus and bacterial corneal ulcer model was established successfully. At 1d after inoculation, 48 eyes were given levofloxacin eye drops when corneal ulcer was confirmed. Then slit lamp inspection and optical coherence tomography(OCT)were performed to measure the central corneal ulcer depth. All the rabbits right eyes were treated with PTK, as an observation group, left eyes were not treated as a control group. The eye section were observed by slit lamp and central thickness of corneal ulcer was measured by OCT at 3 and 7d after this operation. Rabbits were sacrificed and the cornea was removed for pathological section 7d later.

RESULTS: The corneal ulcers in both groups had a tendency to heal, showing a decrease in ulcer area and smoothness of the surface. There was no significant difference in the depth of corneal ulcer between the observation group and the control group before PTK(t=0.706, P=0.484). The difference between the two groups of eyes at 3 and 7d after PTK was obviously(P<0.05).

CONCLUSION: PTK can effectively cure rabbit Staphylococcus aureus corneal ulcer and promote ulcer wound healing, which may be used for clinical treatment of patients with bacterial corneal lesions.]]> 2018/4/24 14:27:16 Xin Zhou, Dong-Yi Yu and Da-Wei He 92true METHODS:Obtained the limbal tissues from rabbits,used tissue block and enzyme digestion method to culture the corneal limbus stem cells in vitro.The growth characteristics of the cultured cells in vitro were observed under inverted microscope.By means of HE,the morphology and construction features of cells were observed.And immunohistochemical method was used to identify the cultured cells.

RESULTS:Rabbit corneal limbus stem cells could be fast and simply cultured by using tissue block and enzyme digestion method.The dynamic observation under microscope showed that rabbit corneal limbus stem cells grew well with a higher proliferative capacity.In HE staining,the morphology and structure of cells were normal.AE5 and P63 cellular immune identification were positive.

CONCLUSION: Tissue block and enzyme digestion method could be a simple and efficient mode for the primary culture of rabbit corneal limbus stem cells.]]> 2018/3/26 15:30:53 Juan Zhao, Dong-Mei Zhan, Mo-Chi Yang and Xin Fan 91true METHODS: Male Kunming mice were randomly divided into the multiple administration group and the single administration group. The multiple administration group were given diacerein eye drop every 2min(3 times in total). The concentrations of the metabolites of diacerein in the cornea were measured by high performance liquid chromatography after given eye drop 5, 15, 30, 60, 120, and 180min. The pharmacokinetic parameters were calculated by pharmacokinetic software(DAS2.1.1).

RESULTS: The metabolites of diacerein, rhein, was detected in the cornea at each time point. The concentration of the metabolite of diacerein in the cornea was 318.678±40.88, 210.02±25.66, 188.83±31.74, 112.24±11.70, 90.28±22.01 and 57.67±13.71μg/g after given eye drop 5, 15, 30, 60, 120, and 180min in the multiple administration group. The concentration in the single administration group was 145.17±19.29, 97.95±10.49, 71.18±18.70, 39.11±2.44, 18.10±2.34 and 9.08±2.04μg/g respectively. The concentration of rhein in the cornea was the highest at 5min after the administration in the two groups. The concentration of the multiple administration group was higher than that in the single administration group at 5, 15, 30, 60, 120, and 180min(P<0.01). The half-life of the drug was 0.89±0.31h in the single administration group.

CONCLUSION: Compared with the single administration, the conjunctival sac multiple administration has the advantages of high drug concentration and long duration. Therefor the conjunctival sac multiple administration is a more effective method to treat acute infectious corneal diseases.]]> 2018/3/26 15:30:53 Ke Yang, Shi-Wei Chen, Xin-Yan Dou, Zhi-Rui Zhang, Xin Jin and Hong-Min Zhang 90true METHODS: Human RPE cell lines(ARPE-19 cell)were treated with different doses of EGF. The methyl thiazolyl tetrazolium(MTT)assay was used to detect cell viability. The 5-bromodeoxyuridine(BrdU)incorporation assay was used to detect cell proliferation and the “scratch-wound assay” was used to detect cell migration ability. The epidermal growth factor receptor(EGFR)and protein kinase B(AKT)proteins were detected by Western blot.

RESULTS:The MTT assay results showed that treatment with 50 and 100ng/mL EGF for 12h increased ARPE-19 cell viability. The BrdU incorporation assay and the “scratch-wound assay” showed that treatment with 100ng/mL EGF for 24h increased ARPE-19 cell proliferation and migration. The Western blot results showed that treatment with 10-100ng/mL EGF for 12h or 100ng/mL EGF for 15-180min increased phosphorylation levels of EGFR and decreased total levels of EGFR. Similarly, treatment with 10-100ng/mL EGF for 12h or 100ng/mL EGF for 15-180min increased phosphorylation levels of AKT, but not affected total levels of AKT.

CONCLUSION: EGF affects ARPE-19 cell viability, proliferation and migration through inducing phosphorylation of the EGFR/AKT signaling pathway. The EGFR/AKT signaling pathway might play an important role in abnormal proliferation and migration of RPE cells in proliferative vitreoretinopathy.]]> 2018/2/27 14:49:15 Xiao-Dong Chen and Jian-Yu Yang 89true METHODS:Sixty healthy SD rats without any eye disease were selected, and the random grouping method was divided into two groups: astragalus injection group(intervention group)and physiological saline group(control group), each group of 30 rats. In each group, 6 were randomly selected for injection of astragalus injection(15mL/kg)and physiological saline(15mL/kg)in the pre-simulated chamber. The rats were sacrificed immediately and removed the eyeballs after the rats were taken out of the simulated module. We observed the changes of retinal morphology with the HE staining method, and determine the retinal SOD and MDA content with colorimetry.

RESULTS:HE staining showed no morphological changes in the two groups of retinas at 2h, and as the time of hypoxia was prolonged, the retinal edema gradually increased, but the intervention group was less edema than the control group. The activity of SOD in both groups decreased with the increase of anoxia time of high altitude, and the comparison of different time points in the group was statistically significant(P<0.05). The content of MDA in both groups increased gradually, and the comparison between different time points in the group was statistically significant(P<0.05). The SOD in the two groups of retinas was significantly different at same time point(P<0.05), except for at 2h without statistical significance. The MDA showed the same situation as SOD.

CONCLUSION: Astragalus injection can reduce the damage degree of retinopathy in rats under the high altitude hypoxia environment, that the mechanism may be related to free radicals, enhance the activity of SOD, reduce the MDA content of lipid peroxides, and enhance antioxidant capacity.]]> 2018/2/27 14:49:15 Xing Pan, Wen-Fang Zhang, Qin Liu, Shu Zhang, Hui-Ling Bai and Xi-Yu Jia 88true METHODS: Siliconized microcapillary tube or Schirmer's strips made in China or in the USA were tested. All three carriers were loaded with equal volume and quantity of bull serum albumin(BSA), fetal bovine serum(FBS)or known concentrations of IL1-Ra, IL-6, MIG, IP-10, MMP9, TIMP1, VEGF and HBD1 mixture and kept at 4℃, -20℃ or -80℃. After a desired time of storage, the proteins were eluted with different elution buffer. Micro BCA^TM protein quantitative kit and enzyme-linked immunosorbent assay kits were used to determine total protein concentrations and the concentrations of the above biological molecules, respectively.

RESULTS: Neither protease inhibitors nor detergent were necessary for protein elution from the Schirmer's strip. When stored at -20℃ and -80℃, three tear carriers showed no differences in the recovery of total protein. The recovery of IL-6 from three carriers under different temperature was similar but the higher than other proteins. The recovery rate of IL-1Ra was 98%±6% from the microcapillary tube and 19%±15% from Schirmer's strip. The recovery rate of HBD1 was 93%±8% from the microcapillary tube and 61%±3% from Schirmer's strip. The recovery rate of MIG was 87%±4% from the microcapillary tube and 69%±4% from Schirmer's strip. Microcapillary tube also had less retention for MMP-9(90%±5%)and TIMP1(103%±7%)compared to Schirmer's strip(62%±3% vs 87%±5%). The strips made in China and in the USA showed similar retention to most of the proteins tested here. When stored at -80℃, the recovery rate for VEGF and IP-10 were 82%±5% and 72%±8%, respectively, significantly higher than that when stored at higher temperature.

CONCLUSION: The selection of tear sample carrier is dependent on target proteins to be recovered. Low temperature deep freezing is a preferred way for tear sample storage. microcapillary tube is applicable for most of the proteins. While for those protein with noticeable polarity, like HBD1, pre-experiment is needed.]]> 2018/1/19 9:47:41 Yu-Qing Rao, Yue Huang and Jing Li 87true METHODS: Totally 50 SD rats were divided into experimental group 40 rats, blank group 10 rats by random number method. The rats in experimental group were randomly divided into 3 groups after the model was successful. Experimental model of high intraocular pressure was sacrificed at 1, 2 and 3wk to observe its pathological structure change and ultrastructure change.

RESULTS: The high intraocular pressure of experimental model in 1, 2 and 3wk all showed the optic nerve and retinal damage. It was to see the optic nerve axon disappear, disorder arrangement of myelin sheath, periodic dissolve or demyelinating degeneration, glial cell proliferation. It showed the cells disordered arrangement of retina, the outer nuclear layer became thick, the inner and outer plexiform layer become thick, the kernel layer became thick, air bubbles, the numbers of ganglion cells reduced, ganglion cells and nerve fiber layer edema, microglia proliferation, vascular membrane capillaries expansion, inflammatory cells appearing. It was to see the retinal ganglion cells layer with microglia proliferation under the electron microscope, ganglion cells structure fuzzy, organelles structures disappear, cell apoptosis, mitochondrial swelling, cytoplasm vacuoles degeneration, membrane plate of outer segment fracture or dissolved. And the damage degree was proportional to the forming time of high intraocular pressure.

CONCLUSION: The morphology change of high intraocular pressure model about the retin and optic nerve proves that it is successful building the model through anterior chamber injection of compound carbomer solution.]]> 2018/1/19 9:47:41 Guo-Long Pang, Fei-Fei Yu and He Sun 86true METHODS: After periocular injection of MPS 10mg, the concentrations for MP and its prodrug conmpound were quantified at different time points using mass spectrum-liquid chromatography in plasma and ocular tissues-slera, choroid and retina, vitreous, iris, aqueous humor, lens and optic nerve.

RESULTS: After periocular injection, the time of maximum concentration(T_max)for MPS in ocular tissues was 0.25 to 1h, in blood plasma was 0.25h. Tmax for its metablite MP in ocular tissues was 0.5 to 6h, in blood plasma was 0.5h. The maximum MPS and MP concentration(C_max)and the area under the curve(AUC_0-t)in ocular tissues from high to low in turn was sclera, optic nerve, the choroid and the retina, iris and lens. The drug concentration of lens was not only the lowest among all tissues, but also much lower than others by far the content, and its mean residence time was the longest.

CONCLUSION: Periocular administration of MPS is an effective way to intraocular drug transmission. It achieves a satisfactory drug distribution in sclera, optic nerve, choroid and retina. It is not easily absorbed by lens.]]> 2018/1/19 9:47:41 Song-Tian Che and Xu Li 85true METHODS: Human retinal vascular endothelial cells were cultured in low glucose or high glucose medium, the genomic DNA was extracted and copy number of mitochondrial DNA was detected by real-time PCR. Effects of resveratrol on the mitochondrial DNA copy in retinal vascular endothelial cells cultured under high glucose medium were studied. The expression of mitochondrial transcription factor A(TFAM)and acetylated proliferator-activated receptor coactivator-1 alpha(PGC-1α)were analyzed by Western blot and coimmunoprecipitation.

RESULTS: High glucose inhibited the copy number of mitochondrial DNA in retinal vascular endothelial cells. However, resveratrol decreased the level of acetylated PGC-1α in retinal vascular endothelial cells, increased the expression of TFAM and the copy number of mitochondrial DNA.

CONCLUSION: Resveratrol may improve mitochondrial function of retinal vascular endothelial cells exposed to a high glucose environment via activation of the PCG-1α-TFAM signaling pathway.]]> 2017/12/18 10:32:18 Yan Wei, Xiao-Qing Su, Hong Li and Bing Wu 84true METHODS: Totally 36 participants 36 male Wistar rats were divided randomly into six groups(6 in every group): normal control group, negative control group, Tianma Gouteng Decoction treatment groups(con-centrations were 0.6g/mL, 1.2g/mL, 2.4g/mL respictively)and ginkgo biloba tablets positive control group(concentrations was 1.2mg/mL). Nothing was done in the normal control group. The optic nerve of right eye in the other groups was done with the optic nerve crush model. Normal control group and negative control group was treated only with water. The average grey scale values of the N-methyl-D-aspartic acid receptor 2B(NMDA2B)receptor protein, beta - amyloid protein(Aβ)in the average grey scale values were detected.

RESULTS: The average grey scale value of Tianma Gouteng Decoction in low, medium and high dose groups about NMDA2B receptor protein was significantly less than that of the negative control group(all P<0.001), and there was no significant difference with the positive control group(P=0.092, 0.411, 0.676), the difference between normal control group and negative control group was significant(P<0.001). The high dose group of beta-amyloid's average grey scale value reduced significantly than the negative control group(P=0.030, 0.001). The low dose group than the negative control group was not obviously(P=0.614). The high dose group was not significantly different from the positive control group(P=0.927), the difference between normal control group and negative control group was significant(P<0.001).

CONCLUSION: Tianma Gouteng Decoction can go through the decrease of the NMDA2B receptor protein expression and the control of beta-amyloid deposition to reduce the retinal ganglion cell injury and apoptosis.]]> 2017/12/18 10:32:19 Fan-Tao Lyu, Yu-Jie Li, Ke Ma and Hai-Yan Wang 83true METHODS: The NTR p75 receptor was used to transfer the retinal pigment epithelium cells as the experimental group, and the non transfected retinal pigment epithelium cells were used as control group. The BrdU test detected the proliferation of two groups of cells. The rate of apoptosis in two sets of apoptosis was measured by PI/in the V-FITC double dye method. The laser microscope detects the ROS levels within the cell. The flow cytometer detected the levels of ROS, mitochondrial markers, cytochrome C expression in RPE cells. The Western blot method detected the expression level of Fas, Caspase-3, and VEGF165 in RPE cells.

RESULTS: The RPE cell proliferation activity was gradually decreasing(P<0.05)with the extension of the p75 NTR receptor transfer time in experimental group. The RPE cell proliferation activity in each transfection point was significantly lower in experimental group than in the control group(P<0.05). The percentage of RPE apoptosis was gradually increased with the extension of transfection time in experimental group(P<0.01). The percentage of RPE cell apoptosis in the experimental group was significantly higher than the control group(P<0.01). ROS fluorescence was significantly better in the experimental group than the control group. Flow cytometry instrument method, according to the results of the experimental group PRE ROS levels in the cell, cytochrome C was significantly higher than control group(P<0.01), RPE cell mitochondria marker levels significantly lower than the control group(P<0.01). The results of the Western blot method showed that the expression levels of VEGF165, Fas and Caspase-3 were significantly higher in the experimental group than in the control group(P<0.01).

CONCLUSION: The over expression of p75 NTR receptor could lead to damage of mitochondria in retinal pigment epithelium cells, but it could also promote the apoptosis reaction, eventually it led to the formation of choroidal neovascularization, so it could be speculated that p75 NTR receptor is the damage factors of retinal pigment epithelium.]]> 2017/12/18 10:32:19 Xin Gu and Xin Yang 82true 2S)on oxidative stress in diabetic cataract rats and its mechanism.

METHODS:SD rats were randomly divided into control group, diabetic group, low-dosage NaHS-treated group, high-dosage NaHS-treated group, and NaHS treated alone group. NaHS was used as a donor of H₂S. The diabetic model was established by a single intraperitoneally administrating streptozotocin(STZ, 65mg/kg). BQ900 slit lamp was used to recorded the changes of lenses. At the end of experiment, the level of superoxide dismutase(SOD)was measured by xanthine oxidase test, the activity of malondialdehyde(MDA)was detected by thiobarbituric acid test, and glutathione(GSH-Px)were detected by corresponding assay kits. Western blotting was applied to detect the expression of SIRT1.

RESULTS:The lenses of diabetic group showed different levels of turbidity, which demonstrated that diabetic cataract model was successfully established. Low-dosage NaHS and high-dosage NaHS treatment dramatically alleviated turbidity levels of lenses in diabetic rats, respectively. Compared to diabetic group, low-dosage NaHS and high-dosage NaHS treatment obviously decreased the levels of MDA and increased the levels of SOD and GSH-Px. Furthermore, the SIRT1 expression in lens of diabetic rats was downregulated, and low-dosage NaHS as well as high-dosage NaHS treatment significantly reversed this change.

CONCLUSION:H₂S protects against oxidative stress in STZ-induced diabetic rats involving upregulation of SIRT1.]]> 2018/11/19 14:44:07 Yan Wang, Chun-Yan Li and Li Yang 81true METHODS: Human retinal vascular endothelial cells were cultured with high glucose medium. Real-time PCR and Western blot were used to detect the expressions of FOXO4 within cells; meanwhile, both FOXO4 RNAi lentivirus and control vector lentivirus were infected the retinal vascular endothelial cells cultured in high sugar culture medium. Real-time PCR and Western blot techniques were used to detect the interference efficiency. After collection of supernatant and cells treated with various interferences, the SOD activity, MDA content in the supernatants and ROS level within cells were detected. Flow cytometry was used to determine the changes of cell apoptosis. Western blot was used to detect the expressions of apoptotic protein cleaved Caspase-3 and cleaved Caspase-9 within cells.

RESULTS: The expression of FOXO4 was increased in retinal vascular endothelial cells after treatment with high glucose medium. FOXO4 RNAi lentivirus infection reduced the expression level of FOXO4 in retinal vascular endothelial cells under high glucose environmental conditions. By contrast, control vector lentivirus had no effect on FOXO4 expression in cells. High glucose induced elevated levels of ROS in retinal vascular endothelial cells, reduced the activity of SOD in cell culture medium, increased the content of MDA, elevated the rate of apoptosis, and promoted the expressions of cleaved Caspase-3 and cleaved Caspase-9 proteins in cells. After down-regulation of FOXO4 expression, retinal endothelial cells were induced by high glucose, the activity of SOD in the cell culture medium increased, the levels of MDA, ROS, apoptosis, and the levels of cleaved Caspase-3 and cleaved Caspase-9 proteins decreased in cells as compared with those of retinal vascular endothelial cells. Moreover, compared with those did not interfere with FOXO4 expression, there was statistically significant differences(P<0.05).

CONCLUSION: High glucose induces the expression of FOXO4 in retinal vascular endothelial cells. Knocking-down of FOXO4 expression reduces oxidative stress and apoptosis induced by high glucose medium.]]> 2018/11/19 14:44:08 Jun Lu, Yu Qin, Wen-Wei Xiao and Mei Zhang 80true METHODS: Forty male SD rats were divided into Group A(normal group), Group B(model group), Group C(Shuangdan Mingmu group)and Group D(positive control group)10 rats(20 eyes)in each group. A rat model of diabetic retinopathy was established by one-time tail vein injection with STZ(50mg/kg). After modeling for 1wk, the rats were given medicine by gavage. After gavage for 4wk rats were sacrificed, and the expressions of VEGF-a, VEGF-b, VEGF-c in the retina tissues were detected by immunohistochemical method.

RESULTS: After gavage for 4wk the average gray values of VEGF-a, VEGF-b and VEGF-c protein in the retina of model group, Shuangdan Mingmu group and positive control group were lower than those of the normal group, and the average optical density were higher than those of the normal group. There was a significant difference between the model group and the normal group(P<0.01). The average gray values of VEGF-a, VEGF-b and VEGF-c expression in Shuangdan Mingmu group and positive control group were higher than those in the model group(P<0.05)and the average optical density value were lower than those in the model group.(P<0.01).

CONCLUSION: Shuangdan Mingmu capsule could significantly reduce the expressions of VEGF-a, VEGF-b,VEGF-c in the retina and had a certain protective effect on the retina of rats in the diabetic retinopathy model.]]> 2018/10/22 14:57:10 Jun Peng, Kun Pan, Zheng-Rong Liu, Yu-Hui Qin and Qing-Hua Peng 79true METHODS:HLEC cell line SRA01/04 cells were cultured in DMEM medium and divided into six groups: control group, galactose treatment group, zinc supplement group, zinc supplementation combined with galactose treatment group, zinc deficiency group, zinc deficiency combined with galactose treatment group. The cell viabilities were assayed by MTT, cell morphology and apoptosis were detected by fluorescence microscope and flow cytometry, respectively.

RESULTS:The cell viabilities induced by galactose(0, 25, 50, 75, 100, 125mmol/L)were(100.0±5.4)%,(97.5±3.2)%,(91.3±5.3)%,(93.4±0.6)%,(86.6±1.4)% and(83.5±0.4)%, respectively. When the concentration of galactose was 100 and 125mmol/L, cell viability was significantly decreased, compared with the untreated cells(P<0.05). Fluorescence microscopy results showed that the cell nucleus remained uniformly stained in control group and zinc supplement group. Nuclear shrinkage, a typical apoptotic morphology, was visible in some cells in the galactose treatment group, zinc supplementation combined with galactose treatment group, zinc deficiency group and zinc deficiency combined with galactose treatment group. The cell apoptosis in the six groups were(1.5±0.1)%,(7.1±0.2)%,(1.4±0.1)%,(4.4±0.2)%,(5.5±0.2)% and(15.8±0.3)%, respectively. The cell apoptosis were significant increased in galactose treatment group, zinc supplementation combined with galactose treatment group, zinc deficiency group and zinc deficiency combined with galactose treatment group, compared with the control group(P<0.05), and those of zinc supplementation combined with galactose treatment group were significant decreased, and zinc deficiency combined with galactose treatment group were increased(P<0.05), compared with the galactose treatment group.

CONCLUSION:Zinc supplementation protects human lens epithelial cells against apoptosis induced by galactose and may have an inhibition effect on cataract formation.]]> 2018/9/14 16:29:44 Yi Jia, Liang-Liang Zhang and Huan Xia 78true METHODS:Primary cultured human RPE cells, the third generation of cells were synchronized with serum-free Dulbecco-modified Eagle medium for 24h, and then grouped: 1)Control group: cultured with 100mg/L BSA for 48h; 2)AGEs-treated group: cultured with 200mg/L AGEs for 48h; 3)PM group: PM1 group: cultured with 16mg/L PM+200mg/L AGEs for 48h; PM2 group: cultured with 32mg/L PM+200mg/L AGEs for 48h. The expression of RAGE protein was detected by immunohistochemistry. The formation of ROS was observed by fluorescence microscopy. The apoptosis of cells was detected by TUNEL.

RESULTS:The expression of RAGE protein, ROS and apoptosis of RPE cells in PM group were significantly lower than those in AGEs-treated group, and decreased with the increase of PM concentration.

CONCLUSION:Pyridoxamine can inhibit the expression of RAGE and the production of ROS, reduce apoptosis, and have a protective effect on RPE cells.]]> 2018/9/14 16:29:44 Min Zhou, Yan Huang, Qiao-Ling Lai and Yun-Ping Deng 77true METHODS: Six-month(n=6)and twelve-month(n=6)old Alzheimer's model mice, together with six-month(n=6)and twelve-month(n=6)old C57 mice were in this experiment. Morris water maze test was to assess the spatial memory. After intragastrical administration of curcumin for consecutive three days, the Aβ plaques in retinas(n=48)from all mice(n=24)were detected by noninvasive in vivo optical imaging.

RESULTS: Morris water maze test: compared with the six-month control group(C6), six-month model mice(AD6)swam longer(P<0.05); and AD6 swam longer distance and experienced more crossing times than C6(P>0.05); compared with the twelve-month control group(C12), twelve-month model mice(AD12)swam longer, swam longer distance(P<0.05)and experienced more crossing times(P>0.05); compared with the six-month model mice(AD6), twelve-month model mice(AD12)swam longer, swam longer distance and experienced more crossing times(P<0.05). Moreover, the result of retinal Aβ plaques: We identified retinal Aβ plaques in postmortem from AD6 and AD12. Two six-month model mice had been detected retinal Aβ plaques, thus the positive rate of retinal Aβ plaques in six-month model mice was 33%; And six twelve-month model mice had been detected retinal Aβ plaques, thus the positive rate of retinal Aβ plaques in twelve-month model mice was 100%; Plaques were undetectable in age-matched non-AD individuals neither in six-month or in twelve-month; the positive rate of retinal Aβ plaques in AD12 was higher than AD6(P<0.05).

CONCLUSION: Six-month model mice(AD6)had already a decline of cognition; and the illness gradually increased with the extension of the disease, and the positive rate of retinal Aβ plaques is increasing with progression of Alzheimer's disease.]]> 2018/9/14 16:29:45 Xi Qin and Yan Lu 76true METHODS: Totally 20 mice were randomly divided into experimental group and PBS group. Erlotinib eye drops was prepared. Erlotinib eye drops and PBS were applied to the two groups of mice at 8, 12, 16 and 20 o'clock each day. OCTA was used to measure the 17 regions of the epithelium and corneal thickness at 1wk, 2wk and 3wk before and after the eye droppings.

RESULTS: There was no significant difference in the thickness of corneal epithelium and cornea between the experimental group and PBS group(all P>0.05). Two weeks after eye dropping, the areas of M, T5, IT5, I5, IN5, N5, T6, IT6, IN6 and N6 in the epithelium and corneal of experimental group were significantly thicker than those of PBS group(all P<0.05). In the third week, 17 areas of epithelium and cornea in experimental group were significantly thicker than those of PBS group(all P<0.05). After treatment with 2 and 3wk in erotinib group and PBS group, there were differences in the average corneal epithelial thickness and the total corneal thickness between each group(all P<0.05). According to the trend analysis of the average change of corneal epithelium and corneal thickness, there were differences between the erotinib group and the PBS group(all P<0.05).

CONCLUSION: Using OCTA, it can be found that ellotini has the effect of thickening corneal epithelium and cornea, and the effect is more obvious with the increase of application times.]]> 2019/8/23 9:51:20 Qing Yuan, Kang-Cheng Liu, Biao Li, Pei-Wen Zhu, Qi Lin, You-Lan Min, Wen-Qing Shi, Lei Ye and Yi Shao 75true METHODS: In this study, 40 females NOD mice were selected. The experimental group consisted of NOD mice that were diagnosed with diabetes while the normal control group consisted of those NOD mice without spontaneous diabetes. Hypodermic injection of Scopolamine hydrobromide(0.5mg/0.2mL)was administered under 40% humidity to the experimental group and placed in a controlled drying box for 12h a day. This was to achieve a dry eye model. Testing indicators on the 1, 7, 10 and 14d after modeling, phenol red thread test was used to measure tear secretion and the eye sections were stained with periodic acid-Schiff(PAS)to examine the morphology and number of conjunctival goblet cells. On the 10d after modeling, the changes in the corneal epithelium were visualized after staining with hematoxylin.

RESULTS:For the NOD mice of the experimental group, the tear secretion was gradually decreased with timing, while there were no obvious changes in the normal control group. The volume of the conjunctival goblet cells of the experimental group became larger, and on the 1d after the molding, the experimental group had decreased density of the goblet cells when compared with the normal control group(P=0.008). From the 7d after the molding, as the time was prolonged, the density of the goblet cells was gradually decreased and the differences between the two group at same time point were significant(all P<0.001). Besides, it was required to observe the corneal epithelium of the two groups on the 10d. The result shows that the corneal epithelium became thinned, some epithelial cells were denatured, and stromal cells became edema.

CONCLUSION: Dry eye model of NOD mice was preliminary established, and the changes of ocular surface were similar to those of dry eye in the clinic.]]> 2019/7/25 14:32:55 *, Chun-Hua Li^*, Zheng-Ri Li, Cheng-Lin Li, Hai-Yan Jin, Ning Ren, Xin-Yu Ru and Ying-Jun Li]]> Hong Cui^*, Chun-Hua Li^*, Zheng-Ri Li, Cheng-Lin Li, Hai-Yan Jin, Ning Ren, Xin-Yu Ru and Ying-Jun Li 74true METHODS:An oxygen analyzer, an electromagnetic valve, and some of polymethyl methacrylate plates were purchased commercially and used for building up the device. A transparent sealed glass box was built with the organic glass and an adhesive. An opening was arranged in front of box for easy insertion and removal of the cage. A vent was arranged on the side. The KY-2F digital display oxygen controller and solenoid valve were placed on the top of the glass box. The oxygen probe and the air supply pipe through the solenoid valve were put into the box from the top of the glass box, Oxygen controller and solenoid valve were connected. Retina sheets and HE histologically staining were used to evaluate the animal model.

RESULTS: Retina with non-perfusion area and neovascularization were observed in the C57BL/6J mice 17^th days after birth from the group in the case with controlled oxygen. Retina with HE staining showed that the neovascularization has penetrated over internal limiting membrane.

CONCLUSION:The presented methodology here for generating mouse case with controlled oxygen is easy to do and apply, the model can truly mimic ROP histologically.]]> 2019/6/21 10:08:46 Hong-Bing Zhang, Bo Zheng, Xiao-Gang Yang and Guang-Qiang Ma 73true METHODS: Thirty rats were divided into blank group, model group and treatment group with 10 rats for each. Rats in the blank group were not treated. Rats in the model group and the treatment group were modeled. Rats in the treatment group were intervened with astragalol. Lens opacity, body mass, body growth and biochemical indexes of rats in each group were detected. The right eyeballs of rats in the three groups were stained with HE.

RESULTS: The rats in the blank group had transparent lens and no opacity; the degree of lens opacity and the overall score of cataract in the treatment group were better than those in the model group(P<0.05); at 3^rd and 5^th weeks, the body weight and length of the rats in the treatment group were higher than those in the model group, and the levels of FPG and 2hPG were lower than those in the model group(P<0.05); after 5wk, the biochemical indexes in the treatment group were significantly improved compared with those in the model group(P<0.05).

CONCLUSION: Astragall can improve the degree of lens opacity in diabetic cataract rats, alleviate the effect of disease on the body weight of rats, regulate the level of blood sugar and biochemical indicators in rats, and alleviate the progress of cataract.]]> 2019/6/21 10:08:46 Jin Xing and Shui-Qing Hu 72true METHODS: Lentivirus carrying the rat ciliary neurotrophic factor(CNTF)coding sequence was transfected into bone marrow mesenchymal stem cells isolated from rat femur. Rats with optic nerve injury constructed by clamp optic nerve method were randomly divided into control group and study group. On the 4, 7, 10 and 13d after successful modeling, the transfused bone marrow were injected into the vitreous cavity of the study group. The control group was injected with the same amount of normal saline. On the 14^th day after successful modeling, the light reflexes of the two groups of rats, the number of retinal ganglion cells and the expression of glial fibrillary acidic protein(GFAP)and Caspase-3 protein were observed.

RESULTS: The recovery rate of the study group was significantly higher than that of the control group(83% vs 25%, P<0.05). The results showed that the retinal cells in the study group were relatively neat. A small amount of vacuoles were observed; the retinal cells in the control group were not well-structured, and obvious vacuoles were observed, and the number of cells was decreased. The results of Western blot showed that the expression level of GFAP and the expression level of Caspase-3 were higher than that of the rats in the study group.

CONCLUSION: Genetic modification induces bone marrow mesenchymal stem cells can effectively treat optic nerve injury in rats.]]> 2019/5/22 14:15:43 Ming Ouyang, Jing-Wen Liu, Ke Liu, Gui-Qin Liu and Bo Qin 71true METHODS: LSCD was induced through corneal alkali burn, C57 mice and New Zealand rabbits were used to establish the animal models. Corneal alkali burn manipulation was accomplished in experimental animals under general anesthesia combined with surface anesthesia in the operated eye. Specifically, mice(n=30)were used to induce complete LSCD model. In brief, the filter paper(diameter of 3mm)that immersed in 1mol/L potassium hydroxide solution was placed on the central corneal surface of the left eye for 30s, followed by washing with saline. In addition, rabbits(n=19)were utilized to establish the partial LSCD model. Briefly, the nictitating membrane(third eyelid)was resected, and the filter paper(diameter of 5mm)that immersed in 1mol/L potassium hydroxide solution was placed on the superior temporal peripheral corneal surface of the left for 30s, followed by washing with saline. After surgery, the model eyes were treated with 0.5% Levofloxacin Hydrochloride Eye Drops four times a day. In addition, the slit-lamp microscope was adopted for observation and photo-taking before burn, as well as at 1, 2, 4wk and 2mo after burn; meanwhile, complications such as corneal ulcer and perforation were recorded. 2mo after surgery, the corneal goblet cell distribution was detected with impression cytology, and the severity of LSCD was classified according to slit-lamp microscopic findings and corneal impression cytology. The animals were sacrificed 2mo after surgery, cornea and conjunctiva sections were made to observe angiogenesis and goblet cell distribution in cornea. Animals died accidentally were not counted into the total number, and the successful induction rates of complete LSCD and partial LSCD models were compared.

RESULTS: Six out of the 30 mice died accidentally, while 2 developed corneal perforation after burn, and the remaining 22 had developed complete LSCD only, yielding the successful induction rate of 92%. 2mo after burn, extensive angiogenesis distribution in the superficial and deep corneal stromal layers could be observed, and pathological sections revealed corneal angiogenesis. Seven out of the 19 rabbits died accidentally, while the remaining 12 had various degrees of LSCD only(partial LSCD, average involving 1.17±0.39 quadrants). Additionally, no corneal perforation was observed, and the successful induction rate was 100%. The result of Fisher's exact test P value is 0.543, without statistical difference. No goblet cells were observed in the normal corneal region, while goblet cells were observed in the LSCD region, with the average density of 58.60±12.58 cell/HP.

CONCLUSION: Central corneal alkali burn can induce complete LSCD; however, some animals will experience failure in model induction due to corneal ulcer and perforation, LSCD is generally serious and may be combined with angiogenesis in deep cornea. Alkali burn in superior temporal cornea can induce partial LSCD, which may be combined with relatively minor corneal lesion, and the corneal angiogenesis is located in the superficial layer.]]> 2019/4/22 9:40:42 Qing-Qin Gao, Ping Wang, Juan Wang, Ming Sun, Ling-Juan Xu, Wei Wang, Hui Zhu, Meng-Lin Jiang, Wei-Kun Hu, Xin-Yu Li and Gui-Gang Li 70true METHODS: Tfb were cultured from Tenon's tissues in POAG patients in vitro. The free-serum DMEM-F12 containing 1.0, 10.0, 100.0ng/mL of CTGF was added into medium for 24h and 48h in different experimental group respectively, and only equal volume of free-serum DMEM-F12 was added in the negative control group. After 24h, the cell proliferation was analyzed through MTT assay, and migration was evaluated by crutch method. After 48h, Semi-quantitative RT-PCR was used to observed the mRNA expression of α-smooth muscl actin(α-SMA), and expression of α-SMA protein was examined by immunochemistry.

RESULTS: The proliferation values A of the cells in 1.0, 10.0, 100.0ng/mL of CTGF group respectively were 0.436±0.009, 0.643±0.009, 0.679±0.006, and 0.423 ±0.156 in the negative control group. The migrated cell number was 34.600±3.507, 70.400±2.074, 80.000±2.915 in different concentrations of CTGF group respectively, and 31.000±3.536 in the negative control group. And also in different experimental groups respectively, the absorbance ratio of α-SMA/β-actin was 0.873±0.161, 1.213±0.312, 1.352±0.376, and 0.851±0.158 in the negative control group, the expressing levels A of α-SMA protein in Tfb were 0.110±0.026, 0.141±0.017, 0.175±0.027, and 0.108±0.020 in the negative control group. The statistics of the above experimental data showed that, comparing with the negative control group, the 10.0 and 100.0ng/mL CTGF groups was statistically significant different(P<0.05), but there was no statistical different between the 1.0ng/mL CTGF group and the negative control group(P>0.05).

CONCLUSION: The proliferation, migration, and phenotypic transformation of Tfb can be promoted in CTGF group in POAG patients. These findings suggest that CTGF may play a role in the development of filtering bleb scarring.]]> 2019/4/22 9:40:43 Xiao-Xia Chen and Yang Cao 69true in vitro.

METHODS: The retinal Müller cells of SD rats were primary cultured by enzyme digestion. The second generation of Müller cells were randomly divided into 7 groups. They were normal glucose concentration(5mmol/L glucose)group, high glucose concentration(25mmol/L glucose)group, high glucose+ berberine group(5, 10, 25, 50 and 100μmol/L). After cultured for 24h, 48h and 72h, the cell proliferative viability was measured by CCK-8 method.

RESULTS: After cultured for 24h, 48h and 72h, compared to the normal glucose concentration group, the absorbance of cells in the high glucose concentration group reduced significantly(All P<0.01). Compared to the high glucose concentration group, the absorbance of cells in different concentration berberine(10, 25, 50 and 100μmol/L)groups increased significantly(All P<0.05). It showed a dose-dependent effect. There was no statistically significant difference on the cells absorbance between high glucose+5μmol/L berberine group and high glucose group(P>0.05).

CONCLUSION: Berberine could reduce the inhibitory effect of high glucose on the proliferative viability of Müller cells to some extent. The intensity of effect was positively correlated with the berberine concentration.]]> 2019/4/22 9:40:43 Yi-Ping Jin and Hao-Hao Zhu 68true + channel of retinal Müller cell membrane cultured in vitro.

METHODS: Human Müller cells were divided into 3 groups(control group, high glucose group and experimental group). The control group were cultured in conventional DMEM medium; the high glucose group were cultured in high glucose DMEM medium; the experimental group were cultured with high glucose DMEM medium and 100μmol/L Aflibercept, and the K⁺ concentration of the cells were detected by MQAE, and the cell survival were detected by MTT assay, the flow cytometry were used to detect apoptosis rates, Western blot analysis were used to detect the Müller cell caspase-3 protein levels.

RESULTS: The Müller cells were positive for glutamine synthetase(GS)after 48h of culture, and the purification degree were above 90%. The relative concentrations of K⁺ in the control group, high glucose group and experimental group were(2.14±0.44)%,(23.11±4.39)%,(5.20±0.92)%, and cell viability were(100.00±0.00)%, respectively(73.24±4.13)%,(85.22±5.33)%, the apoptosis rates were(5.03±1.91)%,(26.73±3.14)%,(16.63±2.73)%, and compared the differences between two groups were statistically significant(P<0.05). Compared with the control group, the level of caspase-3 protein in the high glucose group Müller cells were increased significantly(P<0.05); compared with the high glucose group Müller cells, the caspase-3 protein level in the experimental group Müller cells were decreased significantly(P<0.05).

CONCLUSION: Aflibercept can inhibit the K⁺ channel of retinal Müller cells in vitro, inhibit the apoptosis of Müller cells induced by high glucose, decrease the expression of caspase-3 protein, and promote cell proliferation.]]> 2019/3/22 14:50:40 Qi-Feng Lei and Wei Cai 67true METHODS: The expression of VEGF and CCR3 were detected in 18 patients(18 eyes)with pterygium and compared with the normal conjunctiva epithelium in other 14 cases(14 eyes). The histological differences between the two groups were compared by hematoxylin and eosin(HE)staining. The expression of VEGF and CCR3 in pterygium and normal conjunctival tissues were analyzed by immunofluorescence and immunohistochemical. The mRNA expression of VEGF and CCR3 were assessed by Real-time PCR.

RESULTS: Compared with normal conjunctival tissue, the epithelial layer of pterygium was obvious thickening, the stromal layer was more disordered, and the expression levels of VEGF(0.69±0.0875 vs 0.05±0.0024)and CCR3(0.45±0.0248 vs 0.03±0.0074)were significantly elevated(all P<0.01). The expression level of VEGF mRNA in pterygium was about 12 times that of normal conjunctival tissue(12.33±2.84 vs 1.00±0.08), and the expression level of CCR3 mRNA was about 160 times as many as conjunctival tissue(159.60±34.15 vs 1.00±0.09).

CONCLUSION: The expression of VEGF and CCR3 in pterygium was significantly increased, suggesting that they may be involved in the occurrence of pterygium and promote the development of the disease.]]> 2019/3/22 14:50:40 Hui Liu and Min Zhang 66true METHODS: Blood samples from 15 cases of BD patients and 15 cases of normal control were collected to extracted total RNA in plasma. The miRNAs was labeled, miRNAs array hybridization was performed and then array-scanned and analyzed. We searched verified target genes and selected meaningful miRNAs to underwent real time PCR.

RESULTS: In comparison with the healthy controls, there were 8 anomalous miRNAs, in which 3 miRNAs(hsa-miR-34c-5p, hsa-miR-144-3p, hsa-miR-483-3p)were up regulated and 5 miRNAs(hsa-miR-301a-3p, hsa-miR-224-5p, hsa-miR-454-3p, hsa-miR-17-5p, hsa-miR-199a-5p)were down regulated(all P<0.05).

CONCLUSION: The present examination suggests that aberrant levels of miRNAs could contribute to the pathogenesis of BD. Deviant expression of miRNAs may be involved in the activation of Notch1 and SMAD4 pathway in BD, which could offer a novel therapeutic approach for BD.]]> 2018/12/17 9:54:34 Hui Miao, Xin-Qiao Zhang and Hong Wang 65true in vitro and its influence on the expression of cyclin D.

METHODS: The fresh tissue of human pterygium was cultivated by adherent cell culture in vitro and adherent cells were appraised by immune fluorescence staining. HPF cells were treated with hesperidin(24μmol/L, 48μmol/L, 64μmol/L, 72μmol/L, 96μmol/L, 120μmol/L)and MMC(1.5μmol/L, 7.5μmol/L and 30.0μmol/L). The inhibition rate of cell proliferation was detected by MTT assay 24h, 48h and 72h after treatment, and appropriate concentration and time were selected. The relative expression of cyclin D in HPF was detected by Western blot.

RESULTS: When HPF were treated respectively with hesperidin(48μmol/L, 72μmol/L)and MMC(7.5μmol/L)for 48h, Western blot results showed the relative expressions of cyclin D in blank control group(normal culture), MMC group, hesperidin(48μmol/L)group and hesperidin(72μmol/L)group to be 1.20±0.02, 0.60±0.03, 0.54±0.02, 0.45±0.07(F=73.025, P=0.001)respectively. The relative expressions of cyclin D in MMC group and hesperidin group were lower than that of blank control group(P<0.05); while the relative expressions of cyclin D in MMC group and hesperidin(48μmol/L, 72μmol/L)group showed no significant difference(P>0.05).

CONCLUSION:Hesperidin can inhibit the proliferation of HPF by reducing the relative expression of cyclin D.]]> 2019/11/21 15:09:55 Lin-Lin Liu, Jing Wu, Hui Wang, Jin-Rong Liu, Wei-Lin Wu, Ai-Dong Tang, Yi-Ping Jiang and Cheng-Quan Hu 64true METHODS: The diabetic retinopathy model was established and randomly divided into model group, routine drug group, low dose and high dose picetaxel group with 10 rats each. In the model group, 100mg/kg normal saline was used for gavage, while in the conventional group, 100mg/kg epalrestat was used for gavage. The low dose and high dose picetaxel groups were given 100 and 200mg/kg picetaxel respectively. The retina tissue of five groups of rats was observed by optical microscope, Western blot was used to detect the expression of Bax and Bcl-2 protein, and enzyme-linked immunosorbent assay was used to detect the levels of VEGF, HIF-1 α, ANG Ⅱ, Ang-1, Ang-2 and Tie-2.

RESULTS: The ratio of Bax and Ang-1/Ang-2 in the retina of the high dose group was(1.76±0.05, 3.16±0.09)higher than that of the low dose group(1.01±0.21, 2.98±0.02)(P<0.05). The levels of Bcl-2, VEGF, HIF-1 α, ANG Ⅱ, Ang-1, Ang-2, and Tie-2 in high dose picetaxel group were(0.37±0.06, 121.89±5.45ng/mL, 0.38±0.01pg/mL, 7.58±0.10ng/mL, 8.56±0.04μg/L, 3.24±0.25μg/L, 3.00±0.04μg/L)respectively lower than the lower levels(0.96ng/mL, 0.42±0.02pg/mL, 8.12±0.09ng/mL, 9.10±0.46μg/L, 4.12±0.23μg/L, 3.46±0.15μg/L)(P<0.05).

CONCLUSION: Paclitaxel can inhibit oxidative stress injury and angiogenesis by acting on Ang/Tie receptor signaling pathway, effectively protect retinal tissue of diabetic retinopathy rats in a dose-dependent manner, which provides a theoretical basis for clinical treatment of diabetic retinopathy.]]> 2019/11/21 15:09:56 Dan Guo and Ning Du 63true METHODS: Cornea alkali-burned model was made in 25 rabbits, then animals were grouped and sacrificed at 3d, 7d, 14d, 21d and 28d, respectively. The condition developing of alkali-burned cornea was observed by slit lamp biommicroscopy. The corneas were enucleated for histopathological examination. The infiltration of PMNs was identified by hematoxylin eosin(HE)staining and the expression of MMP-9 was identified by immunohistochemisty in different periods.

RESULTS: The quantity of PMNs and MMP-9 increased on the 3d, reached the lower level on 7d, shown a peak on the 14d, then decreased gradually. The area and depth of corneal stroma after alkali burn were the most severe on the 14d.

CONCLUSION: During the wound healing process, alkali-burned cornea has close relation with the infiltration of PMNs and the expression of MMP-9. The infiltration of PMNs and the expression of MMP-9 is positively correlated in corneal stroma injury after alkali burn.]]> 2019/10/23 15:17:25 Dong-Yu Song, Ming-Hong Gao and Dong-Mei Li 62true METHODS: The T2DM rats successfully modeled were randomly divided into the experimental group(dexamethasone sodium phosphate injection)and the control group(normal saline injection). Each group was randomly divided into three subgroups: peribulbar injection group, post-bulbar injection group and subconjunctival injection group according to different drug delivery routes. Rat tail venous blood was collected 0-24h after injection and blood glucose concentration was determined by dipstick method. After injection, dexamethasone concentration in aqueous aqueous solution of both eyes was determined by HPLC at 0.5-24h.

RESULTS: Dexamethasone sodium phosphate or normal saline injection had no significant effect on blood glucose concentration in T2DM rats(P>0.05). After dexamethasone sodium phosphate was injected in different ways for 0.5-24h, rats in the experimental group received dexamethasone concentration peak under bulbar conjunctiva injection group(957.34±3.60ng/mL)>post-bulbar injection group(859.60±3.56ng/mL)>perbulbar injection group(732.38±4.56ng/mL).

CONCLUSION: Injection of dexamethasone sodium phosphate into bulbous injection, post-bulbar injection and subconjunctival injection had no significant effect on blood glucose of T2DM rats, while subconjunctival injection was simple and could reach higher drug concentration in a short time than that injected into bulbous injection and sub-bulbous injection.]]> 2019/10/23 15:17:26 Yan-Yan Shen, Liang Haung, Li Yan, Xiao-Rong Wu, Yi Shao and Feng Mei 61true METHODS: HLEC were cultured in vitro and then randomly divided into 3 groups. Normal group:normal cultured HLEC; control group: normal cultured HLEC transfected with empty vector; BHMT gene overexpression group(OE): HLEC transfected with BHMT gene overexpression. All groups were cultured in 10% FCS DMEM +5mmol/L Hcy for 16h. After cultured, BHMT mRNA expression was measured by qRT-PCR and Western blot, the cell proliferation was detected by EdU Assay Kit,The level of ROS and GSH of HLEC were measured by Flow Cytometer and Visible Spectrophotometers. The expression level of of protein(GRP78, Nrf2, Caspase-12)was measured by western blotting.

RESULTS: After cultured 16h, cell proliferation ability in OE group was increased by 30.0% compared with NC group(P<0.05).The expression of ROS in normal group(89.2043±0.3511)% was obviously higher than OE group(49.5625±0.4502)%, P<0.05, GSH activity in OE group was obviously higher than control group and normal group,(P<0.05). The expression level of GRP78 in the normal group and the control group was significantly higher than overexpression group. The expression level of Nrf2 in the normal group and the control group was significantly lower than overexpression group. The expression level of Caspase-12 in the overexpression group was significantly lower than that in the control group.

CONCLUSION: BHMT in vitro can prevent the oxidative damage of HLEC by high homocysteine, clear the ROS and decrease the ER stress reaction. Apoptosis of lens epithelial cells was inhibited. BHMT plays an important protective role in oxidative damaged HLEC induced by Hcy.]]> 2019/9/20 14:45:17 Hai-Yan Zhou, Yu-Shun Xue, Hong Yan, Di Li and Xin-Chuan Wang 60true METHODS: All 661W photosensitive cells were treated with 2 000Lx light(48h)and/or different concentrations of Dp(5, 10, 20μmol/L, 24h). Cell activity, intracellular LDH activity, TBARS content and antioxidant enzymes(SOD, GSH-Px, GST)activity were determined respectively. After the healthy SD rats were treated with 3 000 Lx light(24h)and/or Dp \〖100mg/(kg·d)for 4wk\〗, then changes in retinal tissue structure were observed and fluctuations in oxidative stress index(SOD, GSH-Px, GST)were determined.

RESULTS: The results of in vitro experiments showed that the cell activity was significantly decreased after irradiation, the LDH activity and TBARS content were increased, and the activity of antioxidant enzyme system were decreased. However, Dp treatment could increase cell viability, decrease LDH activity and TBARS content, and increase the activity of antioxidant enzyme system. In vivo experiments showed that Dp can protect the structural integrity of retina, reduce the content of TBARS in retinal tissue, and increase the activity of SOD, GSH-Px and GST.

CONCLUSION: Dp may protect retinal against Photochemical factors -induced oxidative damage by regulating the oxidation-antioxidant system.]]> 2019/9/20 14:45:18 Jia-Yuan Peng, Jing-Chang Du, Qian Zhong, Ting-Ting Liu, Li-Qiong Yang, Ai-Lin Wu, Wei Chen, Yan-Feng Zhu and Xiao-Ping Yu 59true METHODS:Totally 40 healthy male SD rats were choose and randomly divided into 4 groups, normal control group, diabetid group, low dose melatonin group and high dose melatonin group. The changes of retinal structure were observed by hematoxylin-eosin staining(HE staining), ultrastructure of retinal ganglion cells were observed by electron microscopy, superoxide dismutase(SOD)and malondialdehyde(MDA)level in the retina tissue were detected by xanthine oxidase method, and protein expression levels of B cell lymphoma/lewkmia-2(Bcl-2), Bcl-2 associated X protein(Bax)and P53 were detected by Western Blot.

RESULTS: HE staining results showed that the rat retina tissue structure of the normal control group was clear, the retinal tissue of diabetes group rats were not clear, the nerve fiber layer was edema, the retina cells of low dose melatonin group were probably distinct, the interlayer cells were arranged neatly, and the inner and outer nuclear layers were slightly disordered, rats retinal structure in high dose melatonin group were further improved than that in low dose melatonin group; Electron microscope results showed that compared with diabetes group, retinal ganglion cells form in melatonin group were improved in different extent; Xanthine oxidase method detected that SOD level in retinal tissue of melatonin group rats were higher than that in diabetic group, the MDA level were lower than that in diabetic group, compared with diabetes group the differences both have statistically significant(P<0.05); Western blot results showed that the protein expression of Bcl-2 in low dose and high dose melatonin group gradually decreased, Bax and P53 protein expressions increased significantly, compared with diabetic group the differences were statistically significant(P<0.05).

CONCLUSION: Melatonin could improve the morphological changes of retina in diabetic rats and inhibit diabetic retinopathy to some extent.]]> 2019/9/20 14:45:18 Hui-Jie Diao, Fang Liu, Jin-Ying Li and Hong Cui 58true METHODS: We made four groups of rabbits for different treatments with dexamethasone and hypertonic solution. All groups were treated with low-permeability solution for 10min to maintain anterior chamber perfusion. Group A was treated with 0.2mL dexamethasone subconjunctival injection, and hypertonic glucose eye drop immediately after surgery. Group B(control group)was subjected to 0.2mL of 0.9% saline subconjunctival injection, and balanced salt solution eye drop immediately after surgery. In group C, 0.2mL dexamethasone subconjunctival injection, and hypertonic glucose eye drop were given on the second day of surgery and in group D(control group), 0.2mL of 0.9% saline through subconjunctival injection and balanced salt solution eye drop were given on the second day of the surgery. The edema degree of cornea was observed with slit lamp before the operation and on the 1, 3, 5 and 7d after operation. The cornea were also examined by anterior segment OCT, and the corneal thickness was measured by A-scan ultrasonography.

RESULTS: In group A, during the entire experimental observation period, the cornea had no edema or only mild edema, the thickness of the central cornea hardly increased, and the number of corneal endothelial cells did not change significantly. There was no significant difference compared with(control group B)before modeling(P>0.05). In groups B, C, and D, corneal edema and corneal thickness increased after the rabbit eyes were modeled. There was a significant difference compared with group A(P<0.05). The number of corneal endothelial cells in the rabbit eyes of groups B and D could not be measured during the observation period due to corneal edema. The number of corneal endothelial cells can be measured in group C up to the 7d after modeling. The number of corneal endothelial cells in group C was significantly reduced as compared with group A(P<0.05), which before modeling and the 7d after modeling.

CONCLUSION: Dexamethasone combined with hypertonic glucose solution has a good protective effect on rabbit corneal endothelial cells. And early application can effectively prevent rabbit corneal edema and this method can also prevent the progression of corneal endothelial decompensation.]]> 2020/8/19 19:18:37 Biao Li, Li Yin and Chao Qu 57true METHODS: The high-glucose micro-environment of diabetic corneal lesions was simulated. After human corneal epithelial cells were resuscitated, cultured and passaged, a normal control group of DMEM complete medium of equal volume of distilled water and a high-glucose treated group of DMEM complete medium containing 25mmol/L glucose were set respectively. After the cells were overgrown, the cells were stimulated with scratches. The growth conditions and changes of the cells in each group were observed and compared under an inverted phase contrast microscope. Western glucose was used to analyze high glucose at different time points(0, 12, 24, 48, and 72h)Cyclin D1 Protein expression in cultured corneal epithelial cells. The qRT-PCR was used to analyze high glucose at different time points and each group Cyclin D1 mRNA expression.

RESULTS: Under the conditions of high glucose treatment in vitro, the repair rate of human corneal epithelial cells was slowed down after injury, floating cells increased, cells reattached less, and cell spacing increased. With the increase of high glucose treatment time, the cell state became worse and the growth rate slow; normal group repaired cell damage faster, increased cell density, regular morphology, and smooth cell membrane. Cyclin D1 expression was up-regulated by Western blot, but the up-regulation effect gradually weakened with time. The highest expression of Cyclin D1 in both groups appeared at 12h. The expression of Cyclin D1 in the high glucose treatment group was lower than that in the normal control group. The qRT-PCR results showed that after high glucose treatment, the expression of Cyclin D1 mRNA was up-regulated, but with the increase of high glucose treatment time, the up-regulation effect weakened, and the mRNA level recovered to the same level as the control group at 48h.

CONCLUSION: In the process of corneal epithelial cell wound healing, high glucose negatively regulates and inhibits the expression of Cyclin D1 protein, and is related to the decline of corneal epithelial cell proliferation and apoptosis.]]> 2020/8/19 19:18:37 Nan-Nan Zhao, Li-Ping Chen, Hao Xiu, Zhen-You Zheng, Ping Tang and Xue-Ying Ji 56true METHODS: High-glucose-induced HRCECs were established by treatment of HRCECs cells with glucose at 30mmol/L for 48h; HG+miR-NC group(transfected miR-NC), HG+miR-221 group(transfected miR-221 mimics), HG+anti-miR-NC group(transfected anti-miR-NC), HG+anti-miR-221 group(transfected anti-miR-221), HG+miR-221+pcDNA 3.1 group(co-transfected miR-221 mimics and pcDNA 3.1), HG+miR-221+pcDNA 3.1-MDM2 group(co-transfected miR-221 mimics and pcDNA 3.1-MDM2), transfected into HRCECs cells by liposome method, and then treated with high glucose; qRT-PCR method for detection the expression of miR-221, p53 and MDM2; the protein expression of p53 and MDM2 were detected by Western blot. The apoptosis of cells was detected by flow cytometry.

RESULTS: Compared with NG group, the expression of miR-221 and p53 was significantly increased, the expression of MDM2 was significantly decreased, and the apoptosis rate was significantly increased in high glucose-induced HRCECs. Overexpression of miR-221 induced apoptosis of high glucose-induced HRCECs cells is more obvious. Inhibition of miR-221 can down-regulate the apoptosis of high glucose-induced HRCECs and down-regulate p53, up-regulate MDM2; overexpression of MDM2 can reverse the inhibition by miR-221 anti-apoptotic effect of cells and regulation of p53 and MDM2 of high-glucose-induced HRCECs.

CONCLUSION: miR-221 can promote the apoptosis of high-glucose-induced human retinal vascular endothelial cells, and its mechanism is related to p53/MDM2 signaling pathway.]]> 2020/8/19 19:18:38 Lu Lu and Huan Li 55true METHODS:Rat retinal vascular endothelial cells were randomly divided into three groups: normal control group, high glucose model group, high glucose model light emitting diode irradiation group, and cells in the high glucose model light emitting diode group began to use light emitting diodes in the incubator 48h after modeling. The cells are irradiated. MTT cell apoptosis experiment was used to detect the apoptosis rate of each group; laser confocal microscope was used to observe the changes of intracellular calcium in retinal vascular endothelial cells; Western blot was used to detect the phosphorylated serine-threonine kinase(P-AKT)protein in each group expression.

RESULTS: The apoptosis rates of normal control group, high glucose model group, and high glucose model light emitting diode irradiation group were 7.54%±2.67%, 31.69%±5.74%, and 21.65%±3.52%, respectively(P<0.05). In the normal control group, the cytoplasm with weak Ca²⁺ fluorescence showed green fluorescence with a pixel value of 192.65±50.54. In the high-sugar model group, the cytoplasm showed a stronger green fluorescence with a fluorescent pixel value of 710.69±100.38. The green fluorescent pixel value was 430.47±80.67, which was significantly higher than the normal control group, but significantly lower than the high-sugar model group. The intra-Ca²⁺ fluorescence pixel values in the three groups were statistically significant(P<0.05). The amount of phosphorylated serine-threonine kinase(P-AKT)protein in these three groups of cells was 10.26±2.47, 2.35±0.16, 7.46±1.64, respectively(P<0.05).

CONCLUSION: High-glucose environment inhibits the activity of threonine kinase pathway, which has an effect on calcium homeostasis of rat retinal vascular endothelial cells and promotes apoptosis. Low-intensity led irradiation can activate threonine kinase pathway and reduce the apoptosis rate caused by high glucose, which is of great application value.]]> 2020/7/22 11:15:57 Feng-Jiu Zhang, Li-Min Zhang, Hong-Jie Wang, Hai-Bo Li, Hai-Ming Wang, Xiang-Dong Peng and Jian-Ling Yang 54true METHODS: Totally 24 rats were randomly divided into control group, operated group, 0.5mg/kg bis(7)-tacrine group and 5mg/kg memantine group. Unilateral elevation of intraocular pressure(IOP)was produced by hypertonic saline injection into an episcleral vein. Animals were orally dosed daily with bis(7)-tacrine or memantine. IOP was measured in both eyes of animals per 3d, and the number of retinal ganglion cells and the thickness of nerve fiber layer axon bundle were measured at 5wk.

RESULTS: Elevated IOP were induced in 3 glaucoma groups. Compared with control group, the retinal ganglion cells decreased from 119.50±8.26 to 79.83±9.58 and the thickness of axon bundle come down from 13.40±0.60 μm to 6.64±0.50 μm in operated group. However the number of the retinal ganglion cells was 109.00±7.04 in bis(7)-tacrine group and 107.33±8.57 in memantine group individually. The thickness of axon bundle was 12.26±0.78μm in bis(7)-tacrine group and 10.13±1.19μm in memantine group individually.

CONCLUSION: Both bis(7)-tacrine and memantine inhibited retinal ganglion cells loss, but only bis(7)-tacrine decreased the thickness declining of axon bundle.]]> 2020/5/25 15:42:59 Jia-Hua Fang, Xiao-Yun Song, Rong Hu, Jun-Xian Liu and Yin-E Xu 53true METHODS: Whole cell patch clamp and intracellular staining with Lucifer Yellow were used in this study to study the morphology and electrophysiological properties of the INL neurons in retinal slices.

RESULTS: Retinal slices prepared in this method possessed a very smooth surface. The cells on the retinal slices maitained very good vitality, and some of the cells even retained their dendritic connections with other cells on the slice. According to the size and location of the cell bodies, neurons in the INL were easy to differentiate. Luciifer Yellow contained in the intracellular solution revealed the morphology of the recorded cell very well. Bipolar cells possessed elongated cell bodies and their processes mainly extended along the vertical direction. Horizontal cells and amacrine cells possessed much bigger and round cell bodies, resided in the outermost and inner most of the INL, respectively. The rest membrane potential and membrane capacitance of horizontal cell and amacrine cell were much higher than that of bipolar cells. Under a voltage step from -60mV to +40mV, 10mV per step, 41.7% of the cone bipolar cells and 64.7% of the amacrine cells exhibited inward sodium current and outward potassium current. Other cells only possessed outward potassium current.

CONCLUSION: The method of preparing retinal slices was very simple, and the viability of the slices were stable. These facilitated the patch-clamp recording of all the neurons in the INL including horizontal cells. Further investigation of the electrophysiological properties of the neurons in the INL was essential in revealing the mechanism of vision.]]> 2020/4/26 11:24:08 Yan-Ming Huang, Mei Yang and Rong-Di Yuan 52true METHODS: Alkali burn model of cornea was established on the right eyes by putting the filter paper with 1.0mol/L NaOH on the center cornea for 1min in 30 white rabbits. The model rabbits were divided randomly into 3 groups after scoring based on Hughes criteria. Normal saline, calf blood deproteinized eye gel and autologous serum eye drops 4 times/day, atropine eye gel 1 times/night, ofloxacin eye gel 1 times/night for 2wk respectively. The morphology of corneal neovascularization was observed on the 7 and 14d, and the area was calculated. On the 14d, the corneas of each group were removed and routine histopathological examinations were performed according to the groups. The concentration of CD45, IL-10, IFN-γ and VEGF in corneal homogenate were determined.

RESULTS:Area of corneal neovascularization: on the 7 and 14d, the area of corneal neovascularization of Group DCS(29.48±2.27, 34.19±2.67mm²), AS(34.19±2.67, 33.89±2.74mm²)(P>0.05). Concentration of CD45, IL-10, IFN-γ, VEGF in cornea homogenate(pg/mL): on the 14th day, the concentration of CD45 Group DCS(0.56±0.04ng/mL), AS(0.54±0.05ng/mL)P<0.05). The concentration of IL-10 Group AS(452.49±11.40pg/mL)>Group DCS>(332.49±13.67pg/mL)>Group Ctrl(111.05±6.95pg/mL)(P<0.05). The Concentration of IFN-γ Group DCS(23.20±2.89pg/mL), AS(22.61±2.72pg/mL)P<0.05). The concentration of VEGF Group DCS(151.14±18.21pg/mL), AS(149.11±14.75pg/mL)P<0.05).

CONCLUSION:AS has the same effect as DCS in inhibiting the release of inflammatory factors(CD45, IFN-gamma and VEGF)and the formation of corneal neovascularization after alkali burn in rabbits, and AS has the strongest effect in promoting the release of anti-inflammatory factors(IL-10)and inhibiting the infiltration of inflammatory cells, followed by DCS.]]> 2020/4/26 11:24:08 Jian Wang, Zheng-Gao Xie, Fang Chen, Jun Zhu and Wei Du 51true METHODS: A Guinea pigs model of form-deprived(FD)myopia were randomly divided into 3 groups: Zhujing formula group, ranibizumab group and saline group(n=20 for each group). Zhujing formula group were fed daily with Zhujing formula solution 3.285g/(kg·d)(1.5mL/d)for 1wk. Ranibizumab group were treated with(intravitreal injections of 0.02mg)ranibizumab at the first day. Saline group were fed with 1.5mL 0.9% saline at the first day. The refraction(Diopter), axial length and choroidal thickness were measured before and at day 1, 3 and 7d postoperative.

RESULTS: The spherical equivalent(SE), axial length and choroidal thickness in ranibizumab group showed no significant trend after intravitreal injection(P>0.05). However, SE and axial length showed trendency to greater myopic shift in the Zhujing formula group and the saline group(P<0.05). The effect began to appear on the first day after administration, achieved the maximum effect after 3d, and faded completely until 7d. On the first day after administration, the diopter and the axial length in ranibizumab group showed the lowest among three groups(P<0.05), and choroid thickness showed the thickest among three groups(P<0.05). At 3d after administration, the diopter and the axial length in the saline group showed the lowest among three groups, and choroid thickness showed the thickest among three groups(P<0.05). There was no significant difference in the parameters among the three groups at 1wk after administration(P>0.05).

CONCLUSION: There is a temperary choroidal thickening of the form deprivation myopia recovery period. The ranibizumab inhibited the thickening of the choroid in the whole recovery period of form deprivation myopia, Zhujing formula slight inhibited the thickening at 3d, and the all change persisted only for 1wk.]]> 2020/3/25 14:45:38 Li-Na Liu, Xing-Wu Zhong, Hong-Shan Liu, Dan-Yang Wang, Jia-Yao Xu, Wei Lao and Ting-Fei Wu 50true METHODS: Totally 36 female New Zealand white rabbits were selected and operated with bilateral ovariectomy and fed for 60d postoperatively to make perimenopausal female rabbit dry eye model(36 eyes, all right eyes). And 24 of them were randomly divided into control group and experimental group(12 for each): the control group used PBS, the experimental group used Mulberry eye drops, the other 12 did not use any eye drops. The Schirmer I tests(SⅠt)and corneal fluorescein(FL)were made, and the tear total protein content, amylase activity, lactoferrin and lysozyme contents and confocal scanning microscopy were performed in two groups before treatment and 2, 4, 6wk after treatment.

RESULTS: There was no significant difference in SⅠt,FL scores,total proteins,lysozyme,lactoferrin contents and amylase activity between two groups at pre-therapy(all P>0.05).After 2, 4 and 6wk of treatment, there were significant changes in SⅠt and FL scores in the two groups(P<0.05). There were no significant changes in lysozyme content, lactoferrin, amylase activity and total tear protein in the experimental group compared with those before treatment(all P>0.05), while there were significant changes in lysozyme content, lactoferrin, amylase activity and total tear protein in the control group(all P<0.05). At 2, 4 and 6wk after treatment, there were differences in the scores of SⅠt and FL, lysozyme content, lactoferrin, amylase activity and total protein content in tear between the two groups(P<0.05). At 6 wk after treatment, the mean number of corneal basal cells and inflammatory cells in control group were 4436±289mm² and 321±91mm²,respectively,which in experimental group were 3219±223mm² and 36±11mm², respectively,there were statistical differences between two groups( all P<0.05). After 6wk treatment, there were no change of corneal nerve bending and less density in control group while the nerve fiber bending and density decreased significantly in experimental group,there were statistical differences between two groups(all P<0.05).

CONCLUSION: Mulberry eye drops has significantly therapy effects on the dry eye induced by decreased androgen and it has a definite clinical application value.]]> 2020/3/13 19:43:57 Wei Jiang, Qian-Min Ge, Biao Li, Qing Yuan, Rong-Bin Liang, Qiu-Yu Li, Pei-Wen Zhu, Jun-Ping Deng, Wen-Qin Shi and Yi Shao 49true METHODS: Thirty healthy adult male SD rats were randomly assigned to the normal control group(CON group, n=10)and the diabetes group(DM group, n=20). After fasting for 12h, the DM group was injected with 1% streptozotocin(STZ)solution, according to 60mg/kg disposable left lower abdominal injection. After 72h, blood was taken from the rat tail vein to detect blood glucose, diabetic model animals were defined as ≥16.7mmol/L. Model rats were randomly divided into diabetes control group(group D)and NAC treatment group(group N). After the model was established, N group of rats were injected with 4μL 1.6μg/μL NAC through the vitreous cavity every week. Rats in CON group and D group were injected with 4μL 0.01mmol/L phosphate buffer saline. All the rats no diet water, group feeding. Body mass and blood glucose were recorded weekly. After the diabetes was modeled, 2mo killed the experimental animals. The thickness of the inner layer of the retina of rats in each group was determined by HE staining. The number of retinal ganglion cells and the level of pigment epithelial derived factor in the retina were measured by immunofluorescence.

RESULTS: The thickness of retinal kernel layer increased in group N compared with group D(P<0.01), and there was no difference between group CON and group(P>0.05). Compared with CON group, the number of retinal ganglion cells decreased in group D(P<0.01), and decreased slightly in group N(P>0.05). Retinal ganglion cells decreased in group D compared with group N(P<0.01). Compared with CON group, PEDF expression decreased in group D(P<0.01), and decreased slightly in group N(P>0.05). The expression of PEDF in group D decreased compared with group N(P<0.01).

CONCLUSION: The protective effect of antioxidant NAC on retinal tissue in early diabetic rats may be due to the up-regulation of PEDF levels in the retina.]]> 2020/3/13 19:43:57 Ling Chen, Yan-Fang Liu and Ping Lin 48true METHODS: Totally 24 healthy SD male rats of 6wk were selected and randomly divided into normal control group(n=6)and experimental group(n=18). The normal control group was fed for 6wk without any intervention. The experimental group was divided into three groups, which were exposed to the array blue light emitting apparatus(455nm-470nm, 391Lx)for 3, 6 and 12h each day for 6wk.

RESULTS: The fundus tissue structure was intact and the cell morphology was normal in the control group. With the extension of blue light irradiation time, the choroid fiber connective tissue of rats in all experimental groups presented hyaline changes, local loose edema, proliferation of small blood vessels, thinning of pigment layer, thinning of cells, gradual reduction of the number of visual cells, and local disappearance. At 3h, the nucleus staining of the experimental group was clear, and no definite changes were observed in the bipolar cell layer and ganglion cell layer. In the 6h and 12h groups, nucleus pyknosis was observed, bipolar cell layer was mildly proliferated, and local cytoplasm was formed in the ganglion cell layer.

CONCLUSION: The photoreceptor cells of retina become thinner, atrophy and disappear with the extension of blue light irradiation time. There was no significant relationship between the injury of pigment epithelial cells and the prolonged exposure time of blue light.]]> 2020/1/19 11:26:19 Hui-Li Li, Zhi-Wei Chen, Xiao-Hong Sun, Guo-Jun Dong, Da-Hong Wang, Sheng-Yan Xiao and Xiao-Dan Li 47true 2O₂-induced apoptosis of human lens epithelial cells.

METHODS: The human lens epithelial cell line HLE-B3 was cultured and divided into normal group, H₂O₂ group(cultured with medium containing 400μmol/L H₂O₂)and H₂O₂+siR-SIAH1 group(transfected with SIAH1 interference sequence, followed by cultured with medium containing 400μmol/L H₂O₂)and siR-NC group(transfected with negative control sequence, followed by cultured with medium containing 400μmol/L H₂O₂). The expression of SIAH1 gene in cells was detected by real-time fluorescent quantitative PCR. The apoptosis rate was detected by flow cytometry. The expressions of p38 MAPK, p-p38 MAPK, Bcl-2 and Bax proteins were detected by Western blot.

RESULTS: The relative expression levels of SIAH1 mRNA in the H₂O₂ group, siR-NC group and H₂O₂+siR-SIAH1 group were higher than that in the normal group(P<0.05). The relative expression level of SIAH1 mRNA in H₂O₂+siR-SIAH1 group was lower than those in the H₂O₂ group and siR-NC group(P<0.05). The apoptosis rates in the H₂O₂ group, siR-NC group and H₂O₂+siR-SIAH1 group were higher than that in the normal group(P<0.05). The apoptosis rate in the H₂O₂+siR-SIAH1 group was lower than those in the H₂O₂ group and siR-NC group(P<0.05). The expression levels of p38 MAPK and Bcl-2 proteins in the H₂O₂ group, siR-NC group and H₂O₂+siR-SIAH1 group were lower than those in the normal group, while the expression levels of p-p38 MAPK and Bax proteins were higher than those in the normal group(P<0.05). The expression levels of p38 MAPK and Bcl-2 proteins in the H₂O₂+siR-SIAH1 group were higher than those in the H₂O₂ group and siR-NC group, while the expression levels of p-p38 MAPK and Bax proteins were lower than those in the H₂O₂ group and siR-NC group(P<0.05).

CONCLUSION: Down-regulation the expression of SIAH1 gene could inhibit H₂O₂-induced apoptosis of human lens epithelial cell line HLE-B3, which might be related to inhibition of p38 MAPK signaling pathway activation.]]> 2019/12/20 14:53:54 Qian-Wei Jia, Xiao-Ping Lei, Min-Hong Shui and Wei Wang 46true METHODS: Corneal neovascularization was established in mice by alkali burn. Sixty mice were then randomly distributed into normal group, Gly group and phosphate buffer solution(PBS)group. The mice were treated with subconjunctival injection of 2g/L Gly solution or vehicle alone every other day for 14d. Corneal inflammation and neovascularization were monitored with a slit lamp microscope. At the end of treatment, the corneas were harvested for hematoxylin-eosin(HE)staining as well as immunohistochemical of CD34 and myeloperoxidase(MPO)staining, microvessel density(MVD), neutrophils were then calculated.

RESULTS: At the 7 and 14d, the CNV area of Gly group were 4.16±0.00 and 7.33±0.13mm² respectively, which were lower than those in PBS group(7.58±0.20 and 9.24±0.08mm²; all P<0.05). The HE pathological staining showed that there were no changes in morphology as well as no neovascularization or inflammatory cell infiltration in the cornea of control group. In the Gly group, blood vessels and inflammatory cell infiltration nearly diminished with collagen in normal shape. While in the PBS group, extensive infiltration of inflammatory cells and neovascularization was examed in the corneal stroma. The immunohistochemical CD34 staining performed that the MVD in the Gly group was 11.13±1.46 bars per square millimeter, which was lower than that in PBS group(34.08±2.46)bars per square millimeter(P<0.001). Additionally, the immunohistochemical MPO staining showed that the number of neutrophils in Gly group was 17.50±1.98 cells per 200-fold field of view, lower than that in PBS group(59.56±4.79, P<0.001).

CONCLUSION: Gly can eliminate corneal inflammation and inhibit corneal neovascularization in mice with acute corneal alkali burn, which provides a new idea for clinical prevention and treatment of corneal neovascularization.]]> 2020/11/19 16:34:46 Pei-Hong Wang, Ling-Han Li, Yong-Ying Zhou, Ming Ying, Yu-Chuan Wang, Jing Li and Xuan Li 45true in-vitro investigations of retinopathies.

METHODS: GFP-adenovirus and CNTF-adenovirus were synthesized then used to transfect BMSCs of passage 3. Blank control group(without adenovirus transfected group), negative control group(GFP-adenovirus transfected group), and experimental group(CNTF-adenovirus transfected group)were included in this study. On the 1, 2, 3d post-transfected, ELISA assay was applied to examine CNTF protein-secretion in the supernate.

RESULTS: GFP-adenovirus and CNTF-adenovirus models were successfully established. The CNTF protein levels in the supernate were higher in experimental group than those in the blank control group and negative control group(P<0.05).

CONCLUSION: CNTF-modified BMSCs by adenovirus could efficiently secrete CNTF protein in-vitro.]]> 2020/11/19 16:34:46 Wen Lin and Guo-Xing Xu 44true METHODS:Six-month-old WT and Crybb2^KO mice were selected respectively. The morphological changes of autophagy of lens were observed by transmission electron microscopy. The expression of autophagy related proteins in the two groups were detected by Western blot method.

RESULTS:Compared with the control group, transmission electron microscopy revealed that mitochondria was accumulated and the number of autophagosomes in lens were higher in Crybb2^KO mice. The relative expression of LC3B in Crybb2^KO group was lower(0.09±0.01 vs 0.26±0.05). The P62 protein and p-mTOR(0.64±0.09 and 0.41±0.03)was higher than WT group(0.43±0.07 and 0.27±0.02).

CONCLUSION:The deletion of CRYBB2 may affect the process of lens autophagy by mTOR pathway and lead to cataract formation.]]> 2020/9/17 16:45:30 Qian Gao, Jian-Cui Li, Yu-Ping Duan, Xiang-Mei Yuan and Qian Li 43true in vitro based on retinal microvascular endothelial cells(ECs)and retinal microvascular pericytes(RMPs).

METHODS: The identified ECs and RMPs of third generation to seventh generation were used for research after isolated, purified and cultured. The cells were stained with cell tracer. Then, it were mixed and inoculated on Matrigel by the surface culture method for dynamic observation. The expression of VEGF-A was assessed during angiogenesis.

RESULTS: At 12h of co-culture, RMPs were recruited by ECs and gathered into cell masses with different sizes. At 24h, ECs/RMPs formed a complex 3D vascular spline network. At 48h, the reticular structure disintegrated obviously, and only a small amount of incomplete and simple reticular structure remained. At 72h, the vascular spline cable network disintegrated completely. In the development of 3D model, the expression of VEGF-A increased, but decreased when it degenerated.

CONCLUSION: This study successfully established a 3D model of rat retinal angiogenesis in vitro based on ECs and RMPs.]]> 2021/8/18 21:32:58 Guang-Hui Liu, Tian-Ye Yang, Ya-Jun Hong, Hang Wang, Ming-Dong Pan, Chun Meng and Chao-Yang Xu 42true METHODS: CAFs and NFs were obtained by tissue primary cultured. The cell morphology of the 3^rd generation purified cells were observed under an inverted phase contrast microscope and identified them using their biomarker by immunohistochemistry for CK, VIM, α-SMA and FAP. Cell growth and proliferation were measured by MTT assay. The expression of FAP mRNA and protein in cells was detected by RT-qPCR and Western Blot.

RESULTS: The CAFs of the eyelid was long fusiform or spindle-shaped, with reduced cytoplasmic processes, more cytoplasmic granules, different cell sizes, disordered arrangement, overlapping growth, and loss of contact inhibition. NFs were in the form of extensive shuttle, radial arrangement, the cytoplasmic particles were rare, there was no overlapping growth phenomenon, and no contact inhibition. The proliferation rate of eyelid CAFs was faster than that of NFs. And the expression of FAP mRNA in CAFs was 3.672±0.221, which was significantly higher than that in NFs(1.034±0.024)(P<0.05). In addition, the expression level of FAP protein in CAFs was high, while NFs were not expressed(P< 0.05).

CONCLUSION: There were significant differences in the biological characteristics of CAFs and NFs, such as morphological structure, growth and the proliferation, growth factor expression and so on. It was therefore suggested that the tumor microenvironment of eyelid basal cell carcinoma had changed, and further induced the biological characteristics and function of NFs, and finally transformed into CAFs. In addition, eyelid CAFs expressed higher expression of FAP than NFs, indicating that FAP may be involved in the occurrence and development of tumor cells in tumor microenvironment, which is associated with the invasive growth of CAFs in eyelid.]]> 2021/7/21 22:23:09 Jin-Hui Zhou, Ning Wang, Qiang-Yuan Mu, Zhu Liu and Da-Qing Wang 41true METHODS:The NPC was prepared by solvent method. Experimental cells are divided into seven groups: control group \〖cultured with dimethylsulfoxide(DMSO)\〗, model group(intervention with 200μmol/L t-BHP), nuclear factor erythroid 2-related factor 2(Nrf2)-siRNA group(cell transfection for Nrf2 gene), naringin group(add 200μmol/L t-BHP after pretreatment with 200μmol/L naringin medium), NPC group(add 200μmol/L t-BHP after pretreatment with 200μmol/L NPC medium), Nrf2-siRNA+ naringin group(after 200μmol/L naringin pretreatment, Nrf2 gene interference, then add 200μmol/L t-BHP)and Nrf2-siRNA+ NPC group(after 200μmol/L NPC pretreatment, Nrf2 gene interference, then add 200μmol/L t-BHP). The intracellular levels of superoxide dismutase(SOD), malondialdehyde(MDA)and total antioxidant capacity(T-AOC)were detected, intracellular level of reactive oxygen species(ROS)was detected by DCFH-DA staining method. The mRNA and protein expressions of HO-1, NQO-1, GCL and Nrf2 were detected by real-time PCR and western blot, respectively.

RESULTS:NPC more significantly increased the levels of SOD and T-AOC, reduced the contents of ROS and MDA than naringin in t-BHP-treated ARPE-19 cells. After naringin and NPC pre-protected ARPE-19 cells, the relative expression and protein expression of Nrf2, HO-1, NQO-1 and GCL mRNA were higher than those of the model group and Nrf2-siRNA group. There were statistically significant differences in the relative expression of 4 genes and the expression levels of 4 proteins in the naringin group and the NPC group, the Nrf2-siRNA+naringin group and the Nrf2-siRNA+NPC group. The expression of Nrf2, HO-1 and NQO-1 protein in the Nrf2-siRNA+naringin group was not significantly different than that in the Nrf2-siRNA group. Compared with the Nrf2-siRNA group, the expression of 4 proteins in the Nrf2-siRNA+NPC group was statistically significant, and the effect of NPC was significantly stronger than that of naringin.

CONCLUSION: After naringin forms a phospholipid complex, it can significantly increase the antioxidant capacity in cells and reduce the oxidation level. It up-regulates the expression of Nrf2 and its downstream antioxidant enzymes and phase Ⅱ detoxification enzymes by activating the Nrf2/ARE antioxidative stress pathway to better protect ARPE-19 cells from oxidative damage.]]> 2021/6/24 15:27:59 Jie Ji, Hai-Tao Yu, Chun-Gang Zhou, Ying Tang, Miao-Miao Tang, Qian-Qian Zhou and Xin-Rong Xu 40true METHODS: A total of 27 patients with suspected PIOL who were admitted to the hospital between January 2015 and December 2019 were enrolled in this study according to the inclusion and exclusion criteria. Totally 21 cases of PIOL and 6 cases of uveitis were diagnosed by pathological examination of diagnostic vitrectomy. Results of gene rearrangement and cytokine levels were retrospectively analyzed. Receiver operating characteristic(ROC)curves were used to analyze the diagnostic value of gene rearrangement, cytokines detection and the combination of the two in PIOL.

RESULTS: Of the 21 patients with PIOL, 15 had IhH FR2 monoclonal rearrangement, with a positive rate of 71%(15/21), and 4 were detected with TCRG clonal gene rearrangement. ROC curve analysis showed that the area under the curve(AUC)of gene rearrangement for diagnosis of PIOL was 0.857. Its sensitivity and specificity were 71.43% and 100.00%. Patients with PIOL had significantly higher vitreous humor IL-10 and IL-10/IL-6 levels than those with uveitis, but no statistically significant difference was found in the IL-6 level between the two groups(P>0.05). ROC curve analysis showed that the AUC of IL-10 was the highest for diagnosis of PIOL. With 170.90pg/mL as the cut-off value, its sensitivity and specificity of IL-10 in diagnosing PIOL were 66.67% and 100.00%, respectively. With 1.95 as the cut-off value, the sensitivity and specificity of IL-10/IL-6 ratio in diagnosing PIOL were 52.40% and 100.00%. The AUC, sensitivity and specificity of gene rearrangement combined with cytokines detection in diagnosing PIOL were 0.893, 95.24% and 83.33%, respectively.

CONCLUSION: The sensitivity of gene rearrangement alone is poor in diagnosing PIOL. Combined use of cytokines detection can improve the diagnostic sensitivity and specificity.]]> 2021/6/24 15:27:59 Yong-Chuan Lyu, Jin-Ping Peng, Ying-Chao Guan, Yang Zhang, Shu-Qing Zhang, Ling-Bo Shu and Yong Tao 39true METHODS: Normal newborn Long Evans rats were randomly divided into normal control group and amblyopia model group, each with 16 rats. Both groups of rats were raised in the same environment. The normal control group did not receive any treatment. On the 13d after birth, the amblyopic model group received monocular suture to establish a classic monocular form deprived amblyopic model. Both groups of rats were received Visual evoked potential(F-VEP)detection on the 51d. Samples were taken immediately after the detection. The transmission electron microscope and Image J image analysis software were used to observe and analyse the synaptic density in the V1M of the primary visual cortex of the two groups of rats. Frozen sections of visual cortex were stained by immunofluorescence histochemical staining by bleaching method and the expression of SYN positive neurons was observed and quantitatively analyzed.

RESULTS: F-VEP examination showed that compared with the normal control group, the P2 latency of the deprived eyes in the amblyopic group was significantly long, and the amplitude of P2 wave was significantly lower than that of normal eyes(P<0.05); Transmission electron microscopy results showed that the synaptic density of the bilateral visual cortex of the amblyopic model group was significantly reduced compared with the normal control group(P<0.05), the contralateral visual cortex of the amblyopic eye decreased more significantly(P<0.05); immunofluorescence staining results showed that the brain slices in the visual cortex of the two groups were intact and the tissue structure was clear under the microscope. Compared with the normal control group, the expression intensity of SYN positive neurons in the amblyopic group was significantly reduced(P<0.01).

CONCLUSION: There is structural synaptic plasticity during the critical period of visual development. Monocular form deprivation can reduce the synaptic density, SYN expression and the visual function in the primary visual cortex of the amblyopic rats.]]> 2021/5/20 16:38:35 Qian Li, Ai-Ling Bi, Xiu-Yan Zhang, Li-Wei Zhang, Zhi-Yuan Lu, Xing-Rong Wang and Hong-Sheng Bi 38true METHODS:Immunofluorescence, real-time quantitative polymerase chain reaction and Western blot were used to detect the protein and mRNA expression of related signal pathways during the process of endoplasmic reticulum stress response induced by ATRA in ARPE-19 cells.

RESULTS: With the accumulation of ATRA concentration, the protein and mRNA levels of endoplasmic reticulum stress response marker proteins chop and BiP were significantly increased(P<0.001); in the downstream signaling pathways, perk, eIF2 α, ATF4, IRE1 α and XBP1 were up-regulated(P<0.001), while the expression of ATF6 did not change(P>0.05).

CONCLUSION: Over accumulation of ATRA induces ERS in ARPE-19 cells and activates PERK-EIF2 α-ATF4 and IRE1 α-XBP1 signaling pathways]]> 2021/5/20 16:38:35 Juan Wu, Jun-Wen Zeng and Dong-Mei Cui 37true METHODS: Totally 69 patients 69 eyes with primary pterygium treat in Zhongnan Hospital of Wuhan University and Hankou Aier Eye Hospital from September 2018 to December 2019 were collected by excision of pterygium during surgery(experimental group). At the same time, a total of 69 patients with normal conjunctival tissue of their same eyes were taken as control group during surgery. The relative expression levels of miR-486-3p in the experimental group and the control group were quantitatively detected by RT-PCR. The Targetscan database, miWalk3.0 database and miRDB database were used to predict the potential target genes of miR-486-3p. DAVID database was used to analyse and enrich the function and pathway of the potential target genes of miR-486-3p. The String website performed an interactional analysis of the potential target genes of miR-486-3p.

RESULTS: The relative expression level of miR-486-3p in the experimental group(6.183±1.366)×10^-6 was significantly different from that in the control group(7.930±1.394)×10^-5(P<0.0001). By the prediction of their target genes and bioinformatical analysis, a total of 436 potential target genes of miR-486-3p were found. The biological functions were mainly concentrated in the regulation of RNA polymerase II promoter transcription, vesicle-mediated transport, transcriptional regulation and the regulation of DNA-dependent RNA metabolism. The Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway was mainly enriched in the Axon guidance pathway and lysosomal pathway. And the Axon guidance pathway might play an important regulatory role in the occurrence and development of pterygium. PPI network analysis further elucidated that the key genes of ABL1 and PLXNA1(cell protein receptor A1)play an important role in the Axon guidance pathway for pterygium.

CONCLUSION: MiR-486-3p might be involved in the occurrence and development of pterygium through SLIT(neuro-targeting factor)/Robo(rotatory guide receptor)and SEMA3A(neuro-guiding factor Semaphorin 3A)/ PLXNA1 of Axon guidance pathways, which resulted in the abnormal new blood vessels of pterygium.]]> 2021/5/20 16:38:35 Yu-Ting Xu, Chen Qiao, Si-Ying He, Shi-Qi Dong, Yun-Feng Xu and Ming Yan 36true METHODS:Thirty-two New Zealand rabbits were selected to establish the corneal epithelial defects models. One eye was treated with normal saline(NC group)and the other eye was treated with 20nmol/L rHGH(rHGH group)in a randomized double-blinded way. The corneal healing process was monitored by the corneal fluorescein staining scores at 0h, 24h, 48h, and 72h after the surgery. The central corneal sensitivity was detected by Cochet-Bonnet corneal esthesiometer and the concentrations of inflammatory mediators interlecukin-1α(IL-1α), interlecukin-17(IL-17), interlecukin-21(IL-21), Leptin, matrix metalloproteinase-9(MMP-9)and tumor necrosis factor-α(TNF-α)in collected tears were measured by multiplex antibody microarray.

RESULTS: The corneal epithelial healing rates of the NC group and rHGH group were(62.52±6.73)% and(79.62±10.62)%(P<0.05),(90.56±9.57)% and(98.43±3.65)%(P<0.05)at 48h and 72h postoperatively. The central corneal sensitivity of rHGH group(4.22±0.26)cm was better than that in NC group(3.22±0.42)cm at 48h after surgery(P<0.05). The expressions of TNF-α and IL-1α increased in both groups at each time point after operation, and the expressions in NC group were higher than those in the rHGH group. Both groups had higher MMP-9 concentrations in the tear fluid at 24 and 48h postoperation in comparison with the point before the operation. The MMP-9 expression in NC group was higher than that in the rHGH group at 48h postoperatively. The expressions of IL-21 in NC group were higher than those in the rHGH group at 24 and 48h postoperation in comparison with the point before the operation(P<0.05). No significant differences in tearIL-17 and Leptin were observed between groups before and after surgery(P>0.05).

CONCLUSION: Topical application of rHGH can accelerate the early stage of rabbit corneal epithelial wound healing in vivo.]]> 2021/5/20 16:38:36 Chi Zhang, Gang-Ping Zhao, Ping Wang, Dong-Mei Cheng and Li Xie 35true 2021/4/21 21:12:00 Qian Li, Fei He and Ran Zhang 34true 2021/4/21 21:12:00 Xi Zou, Guo-Hua Deng, Jun Zhang, Shan-Shan Gu, Han Rong, Li-Hua Kang, Yong Wang and Huai-Jin Guan 33true 2021/3/25 20:05:42 Kai Xu, You-Zhi Tang, Li-Na Liang, Jing Zhang, Le Hou, Jie Liang, Qiang Chen and Qun-Ying Ma 32true METHODS: ARPE-19 cells were stimulated by LPS in vitro to construct the inflammatory injury cell model. Primarily, the cells were divided into five groups randomly. The blank group was cultured in complete medium, and the LPS group was stimulated with complete medium containing 10μg/mL LPS for 24h. The low, medium and high concentration LBP groups were incubated with complete medium importing 0.1, 0.5 and 1mg/mL LBP for 24h separately, and then stimulated with complete medium containing 10μg/mL LPS for 24h. We used the CCK-8 method to observe the cell survival rate, real-time fluorescent quantitative PCR to detect the mRNA expression of inflammatory factors and Western blot to test the changes of phosphorylated protein within the signaling pathway of NF-κB/MAPK.

RESULTS: Compared with normal cells, the survival rate of ARPE-19 cells was decreased after the LPS stimulation. With the increase of exogenous LBP concentration, the survival rate of ARPE-19 cell was gradually increased, while the inflammatory factors expression of cytokines IL-1β, IL-6 and MCP-1 were reduced accompany with the phosphorylated proteins(p-p65, P-IκBα, p-JNK, p-ERK and p-p38)of NF-κB/MAPK signaling pathway were decreased.

CONCLUSION: LBP prevents LPS-induced inflammatory response of ARPE-19 by inhibiting the intracellular inflammatory factors and the phosphorylation of the related protein within NF-κB/MAPK signaling pathway.]]> 2021/2/24 14:14:51 Mei Yan, Qian Ma, Ya-Ling Ma, Yuan Liu and Li-Ping Ma 31true in vitro, establishing a dry eye cell model and analyzing relevant inflammatory factors.

METHODS: Rabbit lacrimal gland epithelial cells were in vitro isolated and cultured, and the activity and purity of primary cells were identified by cell proliferation experiment and immunofluorescence experiment. In addition, 0.5 times IC₅₀ of lipopolysaccharide LPS and TNF-α were used respectively to stimulate rabbit lacrimal gland epithelial cells and then establish two dry eye cell models. Finally, through cell proliferation experiment, ELISA and flow cytometry, the biological characteristics of these two dry eye cell models were compared.

RESULTS:After 12h of culture, the primary cells of lacrimal gland epithelial cells basically adhered to the wall of culture bottles; and 48h later, the cells stretched and almost each of them presented a shape of a long triangle. The activity of primary cells of lacrimal gland epithelium was 92%, and the positive rate of marker Pan-rkeratin was more than 90%, which accorded with the experimental requirements. The IC₅₀ of LPS and TNF-α are 20μg/mL and 4.996ng/mL respectively. After 12h of intervention with LPS(10μg/mL)and TNF-α(2.5ng/mL), the cell activity of the two groups was significantly lower than that of control group(P<0.01); compared between these two groups, the apoptosis rate of TNF-α group is higher than that of LPS group(P<0.01). The levers of IL-1β and IL-6 in the cell supernatants of the two groups were significantly higher than those of the control group(P<0.01); compared between the two groups, IL-1β and IL-6 in TNF-α group were significantly higher than those in LPS group(P<0.01). It was suggested that TNF-α was superior to LPS in simulating inflammatory response of dry eye.

CONCLUSION: This study successfully established a relatively simple and rapid rabbit dry eye cell model with high cell purity and stability, which provided a more stable in vitro experimental model for the basic research on the function of rabbit lacrimal gland epithelial cells and dry eye.]]> 2021/1/19 16:56:24 Ling Li, Meng-Chen Zhu, Dian Li, Ting-Ting Yang, Ting Qiu and Jian Wang 30true METHODS: Totally 18 male c57BL mice aged 8-week-old were divided into control group(n=6), 0mg nicotine group(n=6)and 12mg nicotine group(n=6). The histological and ultrastructural changes of retina were evaluated by hematoxylin and eosin(HE)staining and transmission electron microscope(TEM), respectively. Additionally, the expression of Tuj1 and 8-OHdG was examined using immunofluorescent staining.

RESULTS: In comparison with control group, the thickness of whole retina, nerve fiber layer(NFL)and inner plexiform layer(IPL)was significantly decreased in experimental groups(0mg and 12mg nicotine group)(P<0.01), but no significant difference was observed between 0mg and 12mg nicotine group(P>0.05). The dramatically reduced microvilli of RPE cells were also observed in experimental groups using TEM. Furthermore, residual microvilli were shortened. The expression of Tuj1 was decreased in ganglion cell layer(GCL), NFL and IPL, but no significant changes in the number of retinal ganglion cells were shown among three groups(P>0.05). In addition, the increased expression of 8-OHdG was observed in GCL and inner nuclear layer(INL)in experimental groups.

CONCLUSION: E-cigarette can lead to the retinal damages in mice, which might be due to oxidative stress.]]> 2021/1/19 16:56:24 Shen Nian, Ni-Ni Qiao, A-Li Luo, Juan Li, Shan-Wei Wang, Ya-Jing Mi and Guang-Wei Zhang 29true in vitro.

METHODS: HTFs were divided into ACV treatment group and control group; CCK8 was used to detect cell proliferation rate under different concentration gradients, scratch assay was used to detect HTFs migration ability, and flow cytometry was used to detect HTFs apoptosis and cell cycle.

RESULTS: Compared with the control group, the proliferation rate of HTFs in the ACV-treated group(final concentrations were 1.125, 2.25, 3.375, 4.5mmol/L)was significantly reduced(P<0.05)and was concentration-dependent. The scratch closure rate in the ACV-treated group(final concentrations were 4.5mmol/L)was significantly reduced(P<0.05), the apoptosis rate was significantly increased(P=0.0005), the peak value of the cell cycle G0 / G1 increased(P=0.0011), and the S-phase decreased(P=0.0006). The cell cycle is blocked in the G0/G1 phase.

CONCLUSION: Acyclovir can promote cell apoptosis by blocking the cell cycle of HTFs, and inhibit the proliferation and migration of HTFs.]]> 2020/12/22 18:57:45 Jing-Yi Wu, Zhi-Hang Wu, Shu-Ting Li and Yao Liu 28true METHODS: Totally 72 C57BL/6J mice aged 3-week-old were randomly divided into control group 1, model group 1, intervention group 1, control group 2, model group 2 and intervention group 2, with 12 mice in each group. The first three groups were tested for 3wk and the last three groups were tested for 6wk, except for the groups of control 1 and control 2, all the mice used translucent goggles to cover their right eyes for form deprivation. The mice of intervention 1 and intervention 2 were respectively given intragastric administration modified Zhujing pill suspension 0.546g/(kg·d)(0.1mL/d)for 3wk at the beginning and after 3wk of the experiment. Same amount of saline was given to mice in other groups at the same time of modeling. The eye axis was measured before and after the experiment. At the end of the experiment, the eye of mice was taken for HE staining to observe the thickness changes of each layer of retina. Immunohistochemistry(IHC)and western blotting(WB)were used to detect Bcl-2 and Caspase3 expression of protein.

RESULTS: At the end of the experiment, the axis of model group 1 was significantly higher than that of control group 1(P<0.01), the axis of intervention group 1 was significantly lower than that of model group 1(P<0.01), and the axis of model group 2 was significantly higher than that of control group 2(P<0.01), the eye axis of intervention group 2 was significantly lower than that of model group 2(P<0.01); HE staining showed that the thickness of NFL and ONL of model group 1 was significantly thinner than that of control group 1(P<0.01). The thickness of INL of model group 1 was significantly thinner than that of control group 1(P<0.05), and the thickness of NFL, INL and ONL of intervention group 1 was significantly higher than that of model group 1(P<0.05); The thickness of NFL, INL and ONL model group 2 was significantly thinner than that of control group 1(P<0.01); IHC testing showed that the average optical density of Bcl-2 protein in model group 1 was significantly lower than that of control group 1(P<0.05), which in intervention group 1 was significantly higher than that of model group 1(P<0.01), and which in the average optical density of Bcl-2 protein of model group 2 was significantly lower than that of control group 2(P<0.01), which in intervention group 2 was significantly higher than that of model group 2(P<0.01); Caspase 3 protein average optical density of model group 1 was significantly higher than that of control group 1(P<0.01), which in intervention group 1 was significantly lower than that of model group 1(P<0.01), which in model group 2 was significantly higher than that of control group 2(P<0.05), which in intervention group 2 was significantly lower than that of model group 2(P<0.01); WB test proved that the relative expression level of Bcl-2 protein in model group 1 was significantly lower than that of control group 1(P<0.01), which in intervention group 1 was significantly higher than that of model group 1(P<0.01), and which in model group 2 was significantly lower than that of control group 2(P<0.01), which in intervention group 2 was significantly higher than that of model group 2(P<0.01); The relative expression level of Caspase3 protein in model group 1 was significantly higher than that of control group 1(P<0.01), which in intervention group 1 was significantly lower than that of model group 1(P<0.01), the intervention group 2 was significantly lower than that of model group 2.

CONCLUSION: The results show that the modified Zhujing pill can interfere with the pathological changes of retinal thickness thinning in the process of myopia and formed myopia mice by regulating the expression of apoptosis-related proteins Bcl-2 and Caspase3, and alleviating the apoptosis of retinal cells in myopia formation and myopia mice.]]> 2021/11/22 20:59:15 Ya Mo, Guo-Ting Ren, Xi-Yuan Deng, Jie Ma, Shi-Yun Tang, Lyu-Lyu Zhou, Xi-Li Xiao and Qun Huang 27true 2021/10/22 21:57:36 Jie Bai, Jin-Mei Zhang, Li-Ping Liu, Jia-Ni Wang, Wen Sun and Fan Yang 26true +-K⁺-ATPase and acute reversible lens opacification induced by chloral hydrate in mice.

METHODS: Acute reversible lens opacification was induced by intraperitoneal injection of 4% chloral hydrate(400mg/kg)in 3-month-old(young group)and 24-month-old(old group)C57BL/6 mice. The lens opacification was graded at 10, 20, 30, 45, 60, 90, 120 and 150min after chloral hydrate injection. The histopathological changes of lens were observed by hematoxylin eosin staining, and the expression of Na⁺-K⁺-ATPase in lens was detected by immunohistochemistry.

RESULTS: The development of lens opacity is similar in young and old mice after chloral hydrate injection. The lens opacification in the young group appeared earlier, thicker and lasted longer than the old group. HE staining showed that many vesicles appeared in the cortex below lens epithelial cells(LECs), and the structure of superficial lens fiber cells were disordered after chloral hydrate injection. Immunohistochemical staining showed that the expression of Na⁺-K⁺-ATPase was positive in LECs and fibers. The expression of Na⁺-K⁺-ATPase in LECs were weak before chloral hydrate injection and up-regulated 45min after chloral hydrate injection in young and old groups. The up-regulation of Na⁺-K⁺-ATPase was stronger in the old group than in the young group.

CONCLUSION: Age may play a role in the acute reversible lens opacification induced by chloral hydrate in mice. The expression of Na⁺-K⁺-ATPase is involved in lens opacity induced by chloral hydrate.]]> 2021/9/16 22:17:07 Bai Qin, Jun-Fang Zhang, Mei Yang, Liu-Ping Wu, Li-Hua Kang, Guo-Wei Zhang, Hai-Hong Shi and Huai-Jin Guan 25true METHODS: CCH primary bulbar conjunctival fibroblasts were obtained by tissue block adhesion method. The fibroblasts were purified by trypsin differential digestion method. The growth status and morphological changes of fibroblasts in different periods were observed and recorded under inverted microscope. The fibroblasts were identified by immunofluorescence cytochemical staining.

RESULTS: After 24h of CCH conjunctival tissue adherent to the wall, a small number of cells would be seen crawling out around the tissue blocks. The logarithmic phase of cell growth was from the 2-7d. The cells grew fast, with vigorously proliferation, clear outline, uniform distribution, increas in numbers and clear nuclei. From the 9-15d, the cell growth entered the plateau stage, the tissue blocks gradually aged and lost activity. The cells grew slowly, arranged loosely, the volume became larger, the shape became flat, and a large number of granular substances and vesicles were produced in the cytoplasm. Some cells fell off from the bottom of the culture bottle, and large gaps appeared between the cells. After subculture and purification, the size and morphology of fibroblasts were basically the same. Through cell identification, fibroblasts were long spindle, flat star or multi-process spindle, wide in the middle, oval nucleus, relatively small at both ends, with 2-3 slender processes of different lengths extending outward.

CONCLUSION: Primary CCH bulbar conjunctival fibroblasts can be successfully obtained by tissue block adhesion method. When the cells grow to the 8d, they can be digested and passaged to obtain stable and consistent CCH conjunctival fibroblasts.]]> 2022/9/2 14:23:08 Kai Ma, Jiang Liu, Ya-Hui Wang, Bo-Wen Chi Hua and Min-Hong Xiang 24true METHODS: The Tenon capsule tissues were collected from patients who underwent glaucoma surgery in Hebei General Hospital from September 2018 to September 2019. Primary culture of HTFs was performed by tissue block method. The transforming growth factor-β1(TGF-β1)was used to induce HTFs activation that can mimic glaucoma filtration surgery. The cells were treated with K-115 and divided into 4 groups: the control group was treated with dimethyl sulfoxide(DMSO); TGF-β1 group was treated with 10μg/L TGF-β1 for 24h; TGF-β1 +5 K-115 group was pretreated with 5μmol/L K-115 for 2h and then treated with 10μg/L TGF-β1 for 24h; TGF-β1+10 K-115 group was pretreated with 10μmol/L K-115 for 2h and then 10μg/L TGF-β1 was added for 24h. Cell proliferation was observed by cell proliferation experiment. The migration ability of cells was detected by scratch test. The formation of autophagosomes was observed by transmission electron microscopy. Apoptosis was visualized by Hoechst 33342/PI staining.

RESULTS: Cell proliferation experiment revealed that K-115 could inhibit the proliferation of HTFs induced by TGF-β1. Scratch test suggested that K-115 could inhibit the migration of HTFs induced by TGF-β1. Transmission electron microscope results showed that K-115 could enhance autophagy of HTFs induced by TGF-β1. Hoechst 33342/PI staining suggested that K-115 did not induce apoptosis.

CONCLUSIONS: K-115 may regulate the proliferation and migration of HTFs induced by TGF-β1 by increasing autophagy rather than inducing apoptosis.]]> 2022/9/2 14:23:08 Fang Fan, Jing-Jing Tian, Zhi-Hua Zhao, Qing-Min Ma, Ke-Jun Li and Zhi-Yang Jia 23true METHODS: ARPE-19 cells cultured in vitro were divided into four groups: Control group, the glucose at the concentration of 5.6mmol/L in DMEM/F12 medium; HG group, the glucose at the concentration of 30mmol/L was cultured with DMEM/F12 medium; HG+AG3340 group, the cells were pretreated with AG3340 for 12h, and then cultured in DMEM/F12 medium containing 30mmol/L glucose; The mannitol(MA)group, cultured with DMEM/F12 medium of 5.6mmol/L glucose and 24.4mmol/L mannitol, which used as hypertonic control group. The migration ability of cells was detected by wound healing assay, the invasion ability of cells was detected by Transwell assay, and the relative expression levels of MMP-9, MMP-2, fibronectin and collagen Ⅳ were detected by Western blot.

RESULTS: The results of wound healing assay showed that compared with the Control group, the cell migration rate of scratching after 24h and 48h in the HG group was significantly increased(all P<0.001).After pretreated by AG3340, the cell migration rate was significantly lower than that in the HG group(all P<0.01).Transwell assay showed that compared with the Control group, the number of cell invasion in the HG group was significantly higher than that in the Control group(all P<0.001). After pretreated by AG3340, the number of cell invasion was decreased than the HG group(all P<0.01). Western blot results showed that compared with the Control group, the relative expression levels of MMP-9 and MMP-2 of the cells in the HG group were increased, and the relative expression levels of Fibronectin and Collagen Ⅳ were decreased(all P<0.001). Compared with the HG group, the relative expression levels of MMP-9 and MMP-2 protein in AG3340 pretreatment group were decreased, and the relative expression levels of Fibronectin and Collagen Ⅳ were increased(all P<0.05).

CONCLUSION: High glucose induced ARPE-19 cells with enhanced migration and invasion ability, and AG3340 partially reversed this effect, which was related to the inhibition of MMP-9 and MMP-2 expression and the stability of extra-cellular matrix components.]]> 2022/7/27 16:29:02 Lei Zheng, Guo-Ming Zhang, Miao-Hong Chen and Da-Hui Ma 22true METHODS:The ARPE-19 cells were cultured in vitro and treated with 0,40,80 and 160 μg/mL EGCG, respectively. At the proposed time of treatment the morphological changes were detected by hoechst 33258 staining. The apoptosis rate was detected by flow cytometry. The expression of apoptosis-related factors B lymphocytoma-2 gene(bcl-2), BCL2-Associated X protein(Bax),caspase-3 and p53 were detected by quantitative RT-PCR and Western blotting.

RESULTS: Hoechst 33258 staining showed that the ARPE-19 cells with the increase of EGCG drug concentration, the number of apoptotic cells gradually increased and the apoptotic bodies were observed. Flow cytometry showed that the apoptosis rate increased gradually with the increase of EGCG drug concentration. The apoptosis rates at 40, 80 and 160 μg/mL were 4.95%±0.071%, 11.75%±0.075% and 21.25%±0.919% respectively, which was significantly different compared with the control group(2.8%±1.556%)(P<0.01), presented with a drug concentration-dependent. The results of quantitative PCR and Western blotting showed that EGCG could significantly up-regulate the expression of apoptosis-promoting factors Bax, caspase-3 and the mRNA and protein expression of p53, and down-regulate the apoptosis-inhibiting factor bcl-2, all of these showed concentration-dependent effects.

CONCLUSION:EGCG can obviously induce the apoptosis of ARPE-19 cells. The mechanism is related with the inhibition of bcl-2 and increase the expression of Bax, caspase-3 and p53.]]> 2022/7/27 16:29:02 Mei-Yuan Qiu, Zhi-Xiang Ding, He Jin and Jiao-Jiao Jiang 21true METHODS: There were 36 male 8-week-old C57BL/6J mice randomly divided into 6 groups: blank control(no treatment), ONI 1d group(materials were taken at 1d after optic nerve injury), ONI 3d group(materials were taken at 3d after optic nerve injury), ONI 5d group(materials were taken at 5d after optic nerve injury), ONI 7d group(materials were taken at 7d after optic nerve injury), ONI 14d group(materials were taken at 14d after optic nerve injury). The mice optic nerve model was made by optic nerve gripping, and the mRNA and protein levels of Toll like recepter 9 and myeloid differentiation factor 88 in each retinal were measured by RT-qPCR and Western-blot.

RESULTS: The mRNA and protein levels of Toll like recepter 9 and myeloid differentiation factor 88 in the retina of ONI 1d group were not significantly different from those of the blank control group(P>0.05), the mRNA and protein levels of TLR-9 and MyD88 in the retina of ONI 3d group, ONI 5d group, ONI 7d group and ONI 14d group were significantly increased compared with the blank control group, and the differences were statistically significant(P<0.01). Compared with the blank control group, the mRNA and protein levels of TLR-9 and MyD88 in the retina of mice began to increase at ONI 3d(P<0.01), peaked at ONI 5d(P<0.001), and gradually decreased at ONI 7d(P<0.01).

CONCLUSION: Optic nerve injury can activate the expression of TLR-9 and MyD88 in mice retina. TLR-9 and MyD88 may play an essential role in the process of optic nerve injury.]]> 2022/5/30 15:27:43 Xue-Ying Li, Di Li, Li-Ping Chen and Jing Li 20true METHODS: Forty-eight cyanotic blue rabbits were selected to prepare TPVR animal models by making a penetrating eye injury and intravitreal injection of 0.3mL platelet-rich plasma, and were randomly divided into four groups(n=12), in which the vitreous cavity of the control group was injected with 0.1mL saline; The vitreous cavity of the TA group was injected with 0.1mL(1mg/mL)triamcinolone acetonide; The vitreous cavity of the ART group was injected with 0.1mL(20μg/mL)artesunate; 0.1mL(10μg/mL)luteolin was injected into the vitreous cavity of the LU group. The vitreous and retinal proliferation were observed by fundus photography and ocular ultrasound at 1, 2, 3 and 4wk postoperatively. The expression levels of α-SMA and VIM protein in the vitreous fluid of each group of rabbit eyes were detected by Western Blot at 28d postoperatively, and the retinal tissue structure of each group was observed by retinal HE staining.

RESULTS: At 28d postoperatively, the TPVR grading of rabbit eyes in the TA, ART and LU groups were significantly lower than that in the control group(P<0.05), and the TPVR grading of rabbit eyes in the TA group was significantly lower than that in the ART and LU groups(P<0.05). The expression levels of α-SMA and VIM proteins in the vitreous fluid of the rabbit eyes in the TA, ART and LU groups were significantly lower than those in the control group at 28d after surgery(P<0.01). The results of HE staining showed that the arrangement of retinal layers in rabbit eyesin the control group were disordered, severely distorted or locally broken, the structure of each layer were unclear, the anterior membrane was obviously thickened, and the retina was obviously detached; The arrangement of retinal layersin rabbit eyes in the LU group were slightly distorted, inflammatory exudation was visible in front of the retina, and the retina was superficially detached; The structure of retina in rabbit eyes in the ART group were clear, with mild edema and superficial detachment; The structure of retinal layers in rabbit eyes in the TA group were clear, the arrangement was still neat, the retinal folds were locally visible, and there was no retinal detachment.

CONCLUSION: Intravitreal injection of triamcinolone acetonide, artesunate and luteolin were all effective in preventing and treating traumatic TPVR, among which triamcinolone acetonide has the most obvious effect.]]> 2022/5/30 15:27:43 Ling-Dan Wu, Jie Chen, Zi-Yi Wang and Qi-Hua Xu 19true METHODS: The regulatory role of miR-223-3p in Rbpj gene expression was investigated by a dual luciferase expression reporter system. In the present study, 24 female Lewis rats were randomly divided into EAU model group, normal control(NC)group and blank control(BC)group, and each group included 8 rats. The EAU model group was injected with interphotoreceptor retinoid-binding protein(IRBP)emulsion containing Mycobacterium tuberculin H37RA and complete Freund's adjuvant to induce uveitis, while the NC group was injected with an equal volume of emulsion without IRBP peptide. The rats in the BC group received the same volume of sterile saline solution. At 12d after immunization, the spleen, lymph node and eye tissues in both groups were aseptically isolated, and the expression levels of miR-223-3p and Rbpj RNAs were detected by real-time quantitative PCR(Q-PCR); Meanwhile, the expression levels of Rbpj, IFN-γ and IL-17 proteins were detected by ELISA, and the levels of Th1 and Th17 cell lineages in each tissue from each groups were detected by flow cytometry.

RESULTS: The results of dual fluorescein assay indicated that Rbpj was the target gene which regulated by miR-223-3p. At 12d after immunization, compared with NC group, the relative expression levels of miR-223-3p in spleen, lymph node and eye tissues from EAU model rats were 0.33±0.29, 0.11±0.12 and 0.18±0.11, respectively, accompanied by the down-regulated expression, and the differences were statistically significant(all P<0.05); Rbpj mRNA levels were 3.00±0.06, 1.52±0.12 and 3.01±0.34, respectively, and were all up-regulated, while the differences were statistically significant(all P<0.05). Moreover, the differences in miR-223-3p and Rbpj mRNA levels in spleen, lymph node and eye tissues of rats in the blank control group were not statistically significant compared with those in the NC group(P>0.05); ELISA results revealed that the expression levels of RBPJ, IFN-γ and IL-17 proteins in all tissues from EAU rats at 12d after immunization were significantly higher than those in the NC group( all P<0.05), and there was no statistically significant difference in the expression levels of Rbpj, IFN-γ and IL-17 protein in all tissues of rats in the blank control group compared with the NC group(P>0.05); Meanwhile, flow cytometry results showed that the proportions of Th1 and Th17 cell lineages in all tissues from EAU model group were significantly higher than those from the NC group at 12d after immunization, and the differences were statistically significant(all P<0.05). Furthermore,there was no significant change in the proportion of Th1 and Th17 cells in each tissue in the BC and NC groups(all P >0.05).

CONCLUSION: The miR-223-3p can negatively regulate the expression of the transcription factor Rbpj of Notch signaling pathway. The down-regulated miR-223-3p expression in EAU rats can increase the expression levels of Rbpj gene and protein, and aggravate the differentiation of Th1 and Th17 cells and the expression levels of related molecules IFN-γ and IL-17, which in turn affect the development of uveitis.]]> 2022/4/24 18:49:23 Meng-Xian Zhou, Ru-Yi Qu, Xue-Wei Yin, Li-Jie Guo, Yan Qiu and Da-Dong Guo 18true in vivo, and to explore the feasibility of using this material for corneal transplantation.

METHODS: Fifteen healthy and clean New Zealand white rabbits were selected for a self-control study. The right eye of the rabbit was used as the experimental eye and the left eye was used as the control eye. The experimental eyes used FS adhesived double-layer corneal stromal lens as the material for lamellar keratoplasty, and the control eyes did not undergo manual intervention. At 7,14, and 28d after surgery, a hand-held slit lamp was used to observe the cornea of the rabbits and then score the biocompatibility. The corneas of both eyes were taken for histopathological examination by HE staining to observe the corneal recovery at the same time.

RESULTS: Slit lamp observation results showed that by 28d after the operation, the corneal epithelium of the experimental eyes grew well, the degree of corneal transparency was basically restored, the degree of edema was reduced, the growth of neovascularization to the corneal edge was not aggravated, and no rejection reaction such as epithelial and endothelial rejection lines were seen; The control eyes had clear corneas and smooth corneal epithelium. The results of biocompatibility score showed that the degree of corneal implant edema gradually decreased, the transparency gradually recovered, the rejection reaction was less, and the biocompatibility of corneal implants was better in the experimental eyes after corneal transplantation. There were no differences in the degree of corneal transparency, edema and neovascularization growth between the experimental and control eyes at 28d after surgery(P>0.01). The results of histopathological examination showed that by 28d after corneal transplantation, there were 4-5 layers of corneal epithelial cells covering the surface of the implant in the experimental eyes, the corneal collagen was neatly and regularly arranged, no obvious inflammatory cell infiltration was seen in the implant, the boundary between the two lenses disappeared, the interlayer FS was completely absorbed by the organism, the implant was fused with the implant bed, and no obvious demarcation was seen.

CONCLUSION:Using FS pasted double-layer corneal stroma lens as a graft for lamellar keratoplasty has better recovery, less rejection and better biocompatibility, and can be used for lamellar keratoplasty.]]> 2022/4/24 18:49:23 Jia-Yue Ji, Liu-Qing Wei, Zacharia Ackbarkhan and Jing Zeng 17true METHODS: The synthesis and performance analysis of Cur/Chit-DC. The relationship between FITC/Chit-DC and hRPE cells was observed under an inverted fluorescence microscope after treating hRPE cells with FITC(FITC/Chit-DC)and Cur/Chit-DC(FITC/Cur/Chit-DC)for 24h, keeping them in dark for 1, 3 and 5d respectively.

RESULTS: By mixing Cur and Chit-DC, the nanoparticles containing chitosan derivatives were light yellow. The drug release from the nanoparticles reached equilibrium after 96h, and the cumulative drug release amount was 31.6%. After FITC/Chit-DC was treated with hRPE cells for 1d, most of Chit-DC nanoparticles were still located near the cell membrane. After 3d, the nanoparticles gradually converged towards the nucleus and most of them were located around the nucleus. After 5d, it was observed that Chit-DC nanoparticles had entered the nucleus and were partially degraded under the action of intracellular lysosomes. The relationship between Cur/Chit-DC and cellular action is roughly the same as the relationship between Chit-DC and cellular action.

CONCLUSION: Cur can be continuously released from Cur/Chit-DC nanoparticle, which has long-lasting sustained-release function.]]> 2022/2/24 14:17:20 Hai-Sheng Zheng, Yu-Qing Lan, Fang Li, Xiao-Qian Liang, Zhen-Duo Yang and Xing-Wu Zhong 16true METHODS: High glucose-induced human retinal vascular endothelial cells(HRVECs)were used to establish a cell injury model(high glucose group). Experimental groups include high glucose+dapagliflozin low-dose group(1ng/L dapagliflozin), high glucose+dapagliflozin medium-dose group(5ng/L dapagliflozin), high glucose+dapagliflozin high-dose group(10ng/L dapagliflozin), high glucose+dapagliflozin high-dose+pcDNA group, high glucose+dapagliflozin high-dose+pcDNA-FOXO4 group, and normal sugar group(5.5mmol/L D-glucose). Flow cytometry was used to detect the apoptosis rate. The levels of superoxide dismutase(SOD)and malondialdehyde(MDA)were tested with corresponding kits. Western blot assay was used to detect the protein level of FOXO4.

RESULTS: Compared with the normal sugar group, the apoptosis rate(P<0.05), the level of MDA(P<0.05)and FOXO4(P<0.05)were increased, but the level of SOD was decreased(P<0.05)in high-glucose group. Compared with the high glucose group, cell apoptosis rate(P<0.05), the level of MDA(P<0.05)and the protein level of FOXO4 were decreased(P<0.05), but the level of SOD was increased(P<0.05)in high glucose+medium-dose dapagliflozin group and high glucose+high-dose dapagliflozin group. Compared with high glucose+dapagliflozin high-dose+pcDNA group, the apoptosis rate(P<0.05)and the level of MDA(P<0.05)were increased, but the level of SOD was decreased(P<0.05)in high glucose+dapagliflozin high-dose+pcDNA-FOXO4 group(P<0.05).

CONCLUSION: Dapagliflozin could inhibit oxidative stress and cell apoptosis in high glucose-induced HRVECs by down-regulating FOXO4, thereby reducing cell damage.]]> 2022/2/24 14:17:21 Yang Wang, Ke Wang and Bao-Lan Liu 15true METHODS: ARPE-19 cells were cultured in vitro and divided into two groups: Control group(Control)and drug plus group(PTX). ARPE-19 cells were treated with different concentrations of PTX(0.005, 0.05, 0.5, 5mg/L)for a certain period of time(12, 24, 36, 48, 72h), and CCK8 assay and flow cytometry were used to detect the effects of drug on proliferation and apoptosis of ARPE-19 cells at different concentrations and time points. The same time, the cell cycle was detected by flow cytometry. Morphological changes of cells were observed by immunofluorescence. Expressions of apoptosis-related proteins and barrier function-related proteins were detected by Western blot. The effect of the drug on the cell barrier was measured by measuring the transepithelial resistance of the cells.

RESULTS: PTX reduced the proliferation ability of ARPE-19 cells. After 36h of treatment with low concentration of 0.005mg/L paclitaxel, cell proliferation began to be affected. At the same time, PTX accelerated cell apoptosis was dependent on drug concentration and time. Flow cytometry showed that the cells were arrested in the G2-M phase. In addition, PTX causes significant morphological changes in cells, with normal cells fusiform or irregular. In the PTX group, the number of cells decreased and the cell shape tended to be round. PTX affected retinal barrier function, and the transepithelial resistance of cells was significantly decreased after treatment, and the expression of tight junction proteins ZO-1 and Occludin were significantly decreased compared with the control group(P<0.05). The expression levels of Cleaved-caspase-3 and Bax were significantly increased compared with the control group, while the expression levels of Bcl-2 were significantly decreased(P<0.05)and was dependent on drug concentration and time.

CONCLUSION: PTX can affect the proliferation and apoptosis of ARPE-19 cells, and it depends on time and concentration. In addition, PTX affected the cell cycle and morphology of ARPE-19 cell. At the same time PTX can destroy the barrier function of the retina,suggesting that anti-tumor drugs have a potential toxic effect on the retina.]]> 2022/1/27 16:19:50 Ya-Ting Ye, Guo-Rui Dou, Tian-Fang Chang, Yu Sun, Ya-Li Niu, Zi-Yi Zhou and Zhao-Jie Chu 14true METHODS: Human corneal epithelial cells(HCECs)model was created by osmotic pressure at a concentration of 500mOsm/L for 24h. Serum of rats containing drugs in the blank group, model group, Western medicine group, and Qishen recipe low-dose, medium-dose and high-dose groups were treated on the modeled DE HCECs, and the effects of different drug stimulation on the survival rate of HCECs were tested by CCK-8 method. The expressions of inflammatory factors TNF-α, IL-6 in extracellular fluid were explored by ELISA. The expression of apoptosis factors caspase 1 and AQP5 were detected by immunocytochemistry(ICC). The expressions of AQP5, JNK1, p-JNK1 of HCECs after intervention with different drug concentrations were found by Western blotting.

RESULTS: Compared with the blank group, the survival rate of HCECs in each group was significantly reduced(P<0.01). The extracellular fluid inflammatory factors TNF-α, IL-6 and caspase-1, p-JNK1, AQP5 protein expression levels increased significantly in each group(all P<0.01); In comparison to the model group, the survival rate of HCECs in each medication group increased significantly(all P<0.01). The expression levels of TNF-α, IL-6 in the extracellular fluid of each drug group, AQP 5 and p-JNK1 protein expression in HCECs, and the expression of caspase-1 and AQP5 protein in the western medicine group and the Qishen recipe high and medium dose group were all reduced(all P<0.05). Compared with the western medicine group, the survival rate of HCECs in the Qishen prescription high-dose group was significantly increased(P<0.01). The expression levels of TNF-α and IL-6 in each dose group of Qishen recipe were reduced(all P<0.05), while the expression levels of caspase-1 in the high-dose Qishen recipe group and the AQP5 protein expression levels of the high and medium-dose Qishen recipe group saw a decrease(all P<0.05). However, there was no statistically significant difference in the JNK1 protein expression of HCECs of all the groups detected by Western blotting method(P>0.05).

CONCLUSION: Qishen recipe can not only reduce the JNK1 phosphorylation and AQP5 protein expression of HCECs induced by hypertonicity, but also reduce the expression of inflammatory factors TNF-α, IL-6 and the apoptotic factor caspase-1 of HCECs in the extracellular fluid, thus effectively Inhibit inflammation and apoptosis.]]> 2021/12/21 20:43:50 Lei Zhao, Xiao-Wei Yang, Hui-Min Zhou and Tao Zuo 13true 3O₄@ADM), and observe its ability to target retinoblastoma Y79 cells and its three-mode imaging ability of in vitro ultrasound/photoacoustic/magnetic resonance.

METHODS: FA-PFH-Fe₃O₄@ADM was prepared by the double emulsification method, and its basic characteristics were tested. Test the cellular safety of nanoparticles. Observe the heating phenomenon and phase transition of nanoparticles under 808nm laser. Y79 cells in logarithmic growth phase were divided into four groups. The non-targeted group and the single magnetic target group were added with PFH-Fe₃O₄@ADM nanoparticles(0.625mg/mL). The single folic acid target group and folic acid-magnetic dual target group were added with FA-PFH-Fe₃O₄@ADM nanoparticles(0.625mg/mL). A 4T magnet was placed at the side of the orifice plate of the single magnetic target group and the folic acid-magnetic double target group to evaluate the targeting ability of nanoparticles on Y79 cells. In addition, the ability of FA-PFH-Fe₃O₄@ADM nanoparticles ultrasound/photoacoustic/magnetic resonance three-mode imaging in vitro was observed.

RESULT: FA-PFH-Fe₃O₄@ADM nanoparticles were successfully prepared with uniform distribution and uniform spherical structure. The average particle size of nanoparticles was 338.6±2.20 nm. ADM and Fe₃O₄ were successfully loaded, with an encapsulation efficiency of(41.76±4.12)% and(59.06±13.63)%, respectively. Nanoparticles had good paramagnetic properties and showed no obvious toxicity to Y79 cells. The phagocytic rate of the folic acid-magnetic dual target group(86.19±0.55)% was significantly higher than that of the single folate target group(43.36±5.91)% and single magnetic target group [(28.58±3.23)%, P<0.05]. The FA-PFH-Fe₃O₄@ADM nanoparticles rapidly heat up and undergo phase transitions under the action of laser. Moreover, they can enhance ultrasound and photoacoustic imaging, and they can negatively enhance magnetic resonance imaging in T₂ mode.

CONCLUSION: FA-PFH-Fe₃O₄@ADM nanoparticles have been successfully prepared in this study, which not only has a good targeting effect on retinoblastoma Y79 cells, but also can achieve ultrasound/photoacoustic/magnetic resonance three-mode imaging in vitro.]]> 2022/11/29 14:58:51 Xing Li, Wen-Di Zheng, Hong-Mi Zou and Ming-Xing Wu 12true METHODS: WERI-RB-1 cells cultured in vitro were divided into control group, 0.04U/mL leech extract group and 0.08U/mL leech extract group. The control group was cultured with complete medium for 48h, and the leech extract group was cultured with 0.04 and 0.08U/mL drug-containing medium for 48h, respectively. The expression level of VEGF in conditioned medium of cell culture was detected by ELISA. The mRNA levels of hypoxia-inducible factor-1a(HIF-1a)and matrix metalloproteinase-9(MMP-9)were detected by RT-PCR. The expression levels of HIF-1a, MMP-9, phosphatidylinositol 3-kinase(PI3K)and human phosphorylated AKT(p-AKT)were detected by Western Blot.

RESULTS: Compared with the control group, the expression of VEGF in the conditioned medium with the concentration of 0.04U/ mL and 0.08U/mL of leech extract was decreased(P<0.05), and the inhibition rate was 32.43% and 38.92%, respectively. The expression levels of HIF-1a and MMP-9 mRNA were significantly decreased(P<0.05). The inhibition rates of HIF-1a mRNA expression of the two leech extracts were 27.64% and 24.75%, respectively. The inhibition rate of MMP-9 mRNA expression was 43.97% and 51.48%, respectively. The protein expression levels of HIF-1a, MMP-9, PI3K and p-AKT were significantly decreased compared with the control group(P<0.05). The inhibition rates of 0.04 and 0.08U/mL leech extracts on the protein expression of HIF-1a, MMP-9, PI3K and p-AKT were 55.81% and 43.85%, 39.49% and 47.23%, 33.27% and 29.83%, 52.07% and 30.21%, respectively.

CONCLUSION:Leech extract may inhibit the expression of VEGF in WERI-RB-1 cells via the VEGF/PI3K/AKT pathway or HIF-1a and MMP-9 factors.]]> 2022/11/29 14:58:51 Yuan-Yuan Li, Yan-Lin Zheng, Jian-Chao Li, Yan-Nian Hui, Shi-Hang Ma, Hui Huang, Fang Wang and Hong-Jie Ma 11true METHODS: The tissues of patients with LGBLEL or orbital cavernous hemangioma(CH)were collected. Proteomics analysis was used for the identification of different proteins. Reverse transcription-polymerase chain reaction(RT-PCR), immunohistochemical staining(IHC)and Western Blotting were employed to verify the changes of the differential proteins in CS signal pathway, in order to identify its role in the pathogenesis of LGBLEL.

RESULTS: The results of proteomic analysis showed that the expression levels of proteins C3, C5, C9 and C1q in CS signal pathway in the lacrimal gland tissues of LGBLEL patients were all changed compared with those of orbital CH patients.The results of RT-PCR showed that the mRNA expression levels of C1qA, C5 and C9 in patients with LGBLEL were significantly higher than those patients with orbital CH. The results of IHC showed that the expression levels of C1qA, C3, C5 and C9 were significantly increased in patients with LGBLEL compared with those patients with orbital CH. The results of Western Blotting showed that the protein expression levels of C1qA, C3, and C9 were significantly increased in patients with LGBLEL compared with those patients with orbital CH.

CONCLUSION: The CS has been shown to participate in the pathogenesis of LGBLEL and its classical pathway may be one of the pathways which plays a role.]]> 2022/10/28 16:28:10 Yu-Yue Jin, Rui Liu, Jing Li, Yuan-Yuan Chen, Qian-Nan Ma, Jian-Min Ma and Yi Ding 10true METHOD: A total of 50 Balb/c newborn mice were randomly divided into blank control group, ovalbumin(OVA)+subcutaneous injection group, OVA+nebulized inhalation group, OVA+gastric group, ragweed pollen(RW)+subcutaneous injection group, RW+nebulized inhalation group, RW+gastric group, house dust mite(HDM)+subcutaneous injection group, HDM+nebulized inhalation group, HDM+intragastric group(n=5 animals/group). Except for the blank control group, mice in each group were individually exposed to the corresponding antigens to induce immune tolerance early in life and stimulated with the corresponding antigens in adulthood. The ocular surface was visualized by anterior segment photography. The relative expression level of conjunctival RANTES and IL-17 mRNA was measured by RT-qPCR and serum IL-17 concentration was measured by ELISA.

RESULTS: Compared with the blank control group, the relative expression level of conjunctiva IL-17 mRNA in RW+gastric group was the highest, and it was the lowest in RW+subcutaneous group(all P<0.05). The relative expression level of conjunctiva RANTES mRNA was the highest in RW+gastric group(P<0.001). Compared with the blank control group, the serum concentration of IL-17 was increased in all treatment groups except OVA+nebulizer group and RW+subcutaneous group(P<0.05).

CONCLUSION: The immune tolerance of allergic conjunctivitis induced by subcutaneous injection of antigen was the most suitable method in the early life of mice.]]> 2022/9/22 16:19:01 Meng-Tian Bai, Yun Li and Zhu-Lin Hu 9true 2023/6/21 14:36:09 Chun-Tao Xing, Qi Yu, Lu-Sha Zhang, Jue Li, Hua Guan, Le Wang and Rong Li 8true METHODS: SRA01/04 cells were divided into experimental group and control group. In the experimental group, cells were irradiated with ultraviolet-B(UVB)for 10min to establish the model of oxidative damage, whereas cells in the control group were untreated. Protein expression profile from the two groups was sequenced by isobaric tags for relative and absolute quantitation(iTRAQ). The filtering criteria that fold change >1.2 and p<0.05 was used to determine the differentially expressed proteins(DEPs). Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)database were utilized for functional enrichment analysis of the top 50 DEPs with either up-regulated or down-regulated significance. Furthermore, Pathway commons software was used to establish the protein-protein interaction(PPI)network.

RESULTS: Overall, 552 DEPs were screened out. A total of 176 DEPs were up-regulated in the experimental group compared with the control group, including HMGB1 and USP1, while 376 DEPs were down-regulated, including POLR2A and POLR2B. GO and KEGG enrichment analysis indicated that the top 50 DEPs with up-regulated or down-regulated significance were involved in various crucial biological processes and signaling pathways. PPI network revealed that oxidative damage repair(ODR)-related proteins might play a key role in UVB-induced oxidative damage.

CONCLUSIONS: The expressions of multiple proteins, especially ODR-related proteins, can be altered in SRA01/04 cells via UVB irradiation. These findings may provide cellular-related insights into the pathogenesis of ARC and into proteins or pathways associated with therapeutic targets.]]> 2023/3/30 16:29:49 Xiao-Juan Chen, Guo-Wei Zhang, Peng-Fei Li, Li-Hua Kang and Huai-Jin Guan 7true 2023/3/2 16:43:33 Guang-Hui Liu, Chang-Xuan Shi, Ya-Jun Hong, Yong-Zheng Zheng, Hang Wang and Chun Meng 6true 2023/3/2 16:43:34 Yu-Meng Hao, Cai-Xia Wang, Jing-Xue Ma, Xue-Jing Li and Qing-Li Shang 5true METHODS: Clean male C57BL/6J mice were intraperitoneally injected with N-methyl-N-nitrosourea(MNU, 60mg/kg)to prepare RP mouse model. Electroretinogram(ERG)and hematoxylin-eosin(HE)staining were performed on 7d after modeling to verify the successful modeling. The expression of endoplasmic reticulum stress-related proteins(IRE1, ATF6, PERK, GRP78, Caspase-12)was detected by IHC staining.

RESULTS: The following proteins, including IRE1, ATF6, PERK, GRP78 and Caspase-12, were positively expressed in retina of RP mouse. The optimal concentrations of the above proteins were as follows: IRE1 antibody concentration was 1:1000, ATF6 antibody concentration was 1:500 and 1:1000(with no difference in positive expression, P>0.05), PERK antibody concentration was 1:1500, GRP78 antibody concentration was 1:200 and Caspase-12 antibody concentration was 1:100, the proteins were well expressed at the above concentrations, and the positive expressions of corresponding proteins were different from those of other antibody concentrations(P<0.05).

CONCLUSION: The optimal concentrations for IHC staining in different proteins of mouse RP models were as follows: the concentrations of endoplasmic reticulum stress-related protein antibodies were 1:1000 in IRE1, 1:500 and 1:1000 in ATF6, 1:1500 in PERK, 1:200 in GRP78, and 1:100 in Caspase-12.]]> 2023/1/4 15:06:19 Xiao-Hong Chen, Wan-Jiao Liang, Shi-Shu Huang, Yan Sun, Xin Luo, Lu Lai, Zhao-Sheng Chi, Mei-Zhu Chen, Yun-Peng Wang and Wei-Ming Yan 4true 2023/10/24 9:37:18 Wen-Jing Wang, Shuai Sheng, Jian-Tao Ren, Xu-Dong Huang 3true METHODS:A total of 24 SD rats(24 eyes)who received penetrating keratoplasty were randomly divided into two groups: PBS group received intravitreal injection of PBS(12 rats, 12 eyes)and FasL inhibitor group(12 rats, 12 eyes). Rejection index was recorded every week and blood samples and lymph node were collected at 1, 3 and 5wk after surgery to analyze the proportions of Treg. Corneal tissue was collected for detecting the expression of Fas and FasL and number of apoptosis.

RESULTS: The expression of Fas, FasL in FasL inhibitor group decreased significantly compared with the PBS group(all P<0.05); Corneal cell apoptosis significantly decreased in FasL inhibitor group, and it was the lowest at 5wk after surgery; Treg numbers in blood and lymph nodes significantly increased in FasL inhibitor group at 3wk after surgery(all P<0.05); rejection index of corneal transplantation in the FasL inhibitor group was significantly lower than that of PBS group(all P<0.05).

CONCLUSION:Intravitreal injection of FasL inhibitors after corneal transplantation could reduce the apoptosis in all layers of cornea, increase the number of Tregs in blood and lymph nodes, and alleviate rejection.]]> 2023/9/19 10:01:15 Qian Cao, Lan Li, Yong Li, Yun-Chuan Li, Ying Zou, Jun-Jun Long, Liu-Yu He, Hao-Wen Liu 2true METHODS: Human lens epithelial cells(HLEB3 cells)were propagated in vitro and then separated into two groups: one exposed to standard oxygen levels, added DMEM culture solution containing 10% FBS(normoxic group)and another subjected to low oxygen levels(hypoxic group). The hypoxic condition was emulated by applying a concentration of 100 μmol/L cobalt chloride(CoCl2)for 6, 12, 24, and 48h. The utilization of immunofluorescence staining enabled the detection of Wnt3a and DKK-1 expressions, along with the expression and localization of β-catenin protein in these groups. The expression of DKK-1 mRNA was discerned by quantitative real-time polymerase chain reaction(qRT-PCR).

RESULTS: Immunofluorescence assays indicated an escalating trend in the Wnt3a and DKK-1 protein expression, which corresponded with the increasing duration of hypoxia. Likewise, an intensified nuclear accumulation of β-catenin protein was observed to be directly proportional to the length of hypoxia treatment. The qRT-PCR demonstrated that the difference in DKK-1 mRNA expression between the normoxic group and the group exposed to hypoxia for 6h was not statistically significant(P>0.05), whereas the DKK-1 mRNA expression of the 12, 24, and 48h hypoxia groups were significantly increased(P<0.001).

CONCLUSION: Hypoxia can activate Wnt/β-catenin pathway in lens epithelial cells and induce the expression of DKK-1, thus regulating the Wnt/β-catenin pathway and affecting the EMT process.]]> 2023/9/19 10:01:16 Yan-Song Li, Yi Sun, Yu-Guang Zhu, Ying-Ying Zhong 1true