Background: Pathological retinal neovascularization is a disease characterized by abnormal angiogenesis in retina that is a major cause of blindness in humans. Previous reports have highlighted the involvement of microRNAs (miRNAs) in retinal angiogenesis. Therefore, we aimed at exploring the mechanism underlying miR203 regulating the progression of pathological retinal neovascularization. Methods: Initially, the mouse model of pathological retinal neovascularization disease was established and the hypoxia-induced human retinal microvascular endothelial cells (HRMECs) were generated. Then, miR-203 and SNAI2 expression in HRMECs and retinal tissues was examined. Subsequently, the effects of miR-203 and SNAI2 on viability, migration, apoptosis and angiogenesis of HRMECs were investigated, with the expression of Bax, Ki67, MMP-2, MMP-9, VEGF and CD34 measured. Finally, the regulation of miR-203 or SNAI2 on GSK-3 beta/beta catenin pathway was determined through examining the levels of phosphorylated p-GSK-3 beta and beta-catenin. Results: Poorly expressed miR-203 and highly expressed SNAI2 were found in HRMECs and retinal tissues of pathological retinal neovascularization. Importantly, overexpressed miR-203 or silencing SNAI2 inhibited viability, migration and angiogenesis but promoted apoptosis of HRMECs, evidenced by elevated Bax expression but reduced expression of Ki-67, MMP-2, MMP-9, VEGF and CD34. Moreover, overexpression of miR-203 was found to repress the GSK-3 beta/beta-catenin pathway by downregulating SNAI2. Conclusion: Collectively, this study demonstrated that overexpression of miR-203 suppressed the angiogenesis in mice with pathological retinal neovascularization disease via the inactivation of GSK-3 beta/beta-catenin pathway by inhibiting SNAI2, which provided a novel therapeutic insight for pathological retinal neovascularization disease.

Diabetic retinopathy (DR) is a devastating complication of diabetes. The aim of the present study is to investigate the exact role and mechanism of long noncoding RNA MALAT1 (MALAT1) in the progress of DR. An oxygen-induced retinopathy (OIR) mouse model and high glucose (HG) stimulated human retinal microvascular endothelial cells (HRMECs) were employed to mimic the pathological statues of DR. Quantitative real-time PCR (qRT-PCR) and Western blot results showed that MALAT1, VEGFA, and HIF-1α levels were increased in DR retinal tissues and HG-stimulated HRMECs, whereas the expression of miR-203a-3p was decreased. Knockdown of MALAT1 or upregulation of miR-203a-3p both suppressed HG-induced proliferation, migration, and tube formation of HRMECs. A dual-luciferase reporter assay showed that miR-203a-3p could bind to the predicted seed regions of MALAT1 as evidenced by the reduced luciferase activity. Furthermore, enforced downregulation of miR-203a-3p abolished the suppressive effect of MALAT1 silencing on HRMEC cell migration and tube formation. In conclusion, these data demonstrated that MALAT1 may affect angiogenesis by sponging miR-203a-3p in DR, suggesting that MALAT1 may act as a novel therapeutic target for the treatment of DR.

Prupose This study aims to explore risk factorsassociated with ocular ischemia syndrome (OIS) in patients with late stage carotid artery occlusion (CAO). Methods Eighty-seven patients with CAO were enrolled and examined via patient history questionnaire, carotid duplex ultrasonography,transcranial doppler sonography (TCD) and eye examination (slit lamp, ophthalmoscopes). Results Twenty-two patients were diagnosed with OIS. Female patients are more likely to development OIS (OR=2.78). Patients without collateral circulation were associated with risk of OIS (OR=4.245, P<0.05). Conclusions One quarter of patients with CAO patients had OIS and sex and non-collateral circulation building were associated with OIS.

Proliferative diabetic retinopathy (PDR) is a common complication of diabetes mellitus, characterized by abnormal retinal angiogenesis. MicroRNA-203-3p (miR-203-3p) was found to be down-regulated in a murine model of proliferative retinopathy. This study was performed to explore the role of miR-203a-3p in retinal angiogenesis of PDR. Firstly, a rat OIR model, which was used to mimic PDR, was established and the OIR rats were treated with scrambled control or miR-203a-3p agomir by intravitreal injection. The results showed that the level of miR-203a-3p was decreased in OIR rats, and forced over-expression of miR-203a-3p inhibited OIR-induced retinal angiogenesis as evidenced by reduced blood vessel profiles and CD31 expression. OIR-induced up-regulation of VEGFA, HIF-α, PCNA, and MMPs in the retina was also counteracted by miR-203a-3p. Additionally, high glucose (HG)-induced proliferation, migration and tube formation of human retinal microvascular endothelial cells (HRMECs) were also dampened by the up-regulation of miR-203a-3p. Dual-luciferase reporter assay showed that miR-203a-3p could specifically bind to the 3'UTR of VEGFA and HIF-1α. Over-expression of VEGFA or HIF-1α restored the tube formation activity of HRMECs suppressed by miR-203a-3p. In conclusion, our findings demonstrate that up-regulation of miR-203a-3p might inhibit pathological retinal angiogenesis of PDR by targeting VEGFA and HIF-1α.

Human potassium channel tetramerization domain containing 12 (KCTD12, also known as Pfetin) is a member of the KCTD family which consists of 26 members. It has been reported that KCTD12 regulates agonist potency and kinetics of GABA(B) receptor signaling. Proteomic analysis indicates that KCTD12 may be a potential biomarker for the diagnosis and prognosis of gastrointestinal stromal tumors. However, little has been reported concerning the role of KCTD12 in the other tumor types. In the present study, we designed and subcloned N-terminally Flag-tagged human KCTD12 into the pLVX-Puro vector. We then generated a human uveal melanoma cell line (OCM-1) stably expressing KCTD12. Using this stable cell line, we performed a series of experiments including colony formation, invasion, migration and wound healing assays, flow cytometry and western blotting. Based on the experimental results, we first demonstrated that KCTD12 effectively suppressed the proliferation of OCM-1 cells and limited the spread of OCM-1 cells. In the flow cytometric analysis, prolongation of the progression of G2/M to G1 phase in the KCTD12-overexpressing OCM-1 cells was observed. In addition, inhibition of KCTD12-overexpressing OCM-1 cell xenograft growth in nude mice was observed. Taken together, KCTD12 may serve as a novel therapeutic target for patients with uveal melanoma.

PURPOSE:A meta-analysis was conducted to assess the effects of the aldose reductase (ALR) C-106T polymorphism on the risk for development of diabetic retinopathy (DR) in the Chinese population.MATERIALS AND METHODS:Relevant studies were identified using PubMed, Springer Link, Ovid, Chinese Wanfang Data Knowledge Service Platform, Chinese National Knowledge Infrastructure (CNKI), and Chinese Biology Medicine (CBM) through 21 March 2015. Pooled odds ratios (ORs) and 95% confidence intervals (CIs) were used to assess the strength of the associations.RESULTS:This meta-analysis identified 11 studies, including 1386 DR cases, 1594 diabetes mellitus (DM) control cases, and 472 healthy control cases. In the overall analysis, a non-significant association between the ALR C(-106)T polymorphism and DR was found in the Chinese population. In subgroups stratified by ethnicity, significantly increased risks for DR in association with ALR C(-106)T variants were found in the Chinese Han population. When compared with healthy controls, we found the following associations: T versus C (OR, 1.63; 95%CI, 1.23-2.17), TT versus CC (OR, 2.04; 95%CI, 1.03-4.02), and TT + CT versus CC (OR, 1.82; 95%CI, 1.28-2.57).CONCLUSIONS:Our meta-analysis showed that ALR C-106T variants appear to influence the risk for DR in Chinese Han persons. Studies with larger sample sizes and wider population spectra are warranted to verify this finding.

Objective To investigate the effects of recombinant human Vasostatin on the inhibition of Nd:YAG laser induced mice choroidal neovascularization (CNV). Methods In 30 adult C57BL/6J mice(30 eyes), choroid angiogenesis was induced by Nd: YAG laser fundus photocoagulation in the right eye. The mice were randomly divided into control group, low dose group and high dose group with 10 mice in each group. Immediately after the Nd:YAG laser photocoagulation, retrobulbar injection of 10 μl saline was peformed in all mice in group A ( control group);Mice in treatment group was retrobulbarly injected with 10μl of recombinant human Vasostatin at the concentration of 2μg/μl in group B and 4μg/μl in group C. The choroidal neovascularization was evaluated by fundus fluorescence angiography 7 and 14 days after photocoagulation. Results In the treatment group, the incidence of CNV and spot fluorescein leakage were lighter than that of control group 7 d and 14 d after photocoagulation. Seven days after photocoagulation, choroidal neovascularization incidence was 68% in group A. The incidence rates of choroid neovascularization in group B and group C were 56% and 48% respectively. There was statistically significant difference (P<0. 05). Fourteen days after photocoagulation, the choroid neovascularization incidence was 72% in group A. It was 50% and 38% in group B and group C respectively, which was statistically significant different from group A (P<0. 01). Conclusion Vasostatin can effectively suppress experimental laser-induced mouse CNV. Choroidal injury by the Nd:YAG laser can successfully induce mouse CNV model.

The aim of this study was to be the first to establish a method for determination of rutin content in Qi Ming Granules, providing a scientific basis for the development of its quality standards. Measurement was done directly by RP-HPLC using Agilent TC-C18 chromatographic column with a mobile phase of methanol-water-phosphoric acid (48.5: 51.5: 0.25, pH3.5), flow rate of 1.0 mL/min, and detection wavelength of 360 nm, ruggedness test was performed. The injection amount of rutin had a good linear relationship with the peak area within the range of 0.0300-1.498 mu g, recovery rate was 99.8%, RSD = 1.23% (n = 6). Rutin contents in 10 different batch numbers of Qi Ming Granules were measured to be between 21.09-29.01 mg/g. The method established had the advantages of simplicity, rapidness and high precision, and was used for the first time as a quality inspection method for Qi Ming Granule.

Neural injury is associated with the development of diabetic retinopathy. Müller cells provide structural and metabolic support for retinal neurons. High glucose concentrations are known to induce Müller cell activity. Agmatine is an endogenous polyamine, which is enzymatically formed in the mammalian brain and has exhibited neuroprotective effects in a number of experimental models. The aims of the present study were to investigate whether agmatine protects Müller cells from glucose-induced damage and to explore the mechanisms underlying this process. Lactate dehydrogenase activity and tumor necrosis factor-α mRNA expression were significantly reduced in Müller cells exposed to a high glucose concentration, following agmatine treatment, compared with cells not treated with agmatine. In addition, agmatine treatment inhibited glucose-induced Müller cell apoptosis, which was associated with the regulation of Bax and Bcl-2 expression. Agmatine treatment suppressed glucose-induced phosphorylation of mitogen-activated protein kinase (MAPK) protein in Müller cells. The present study demonstrated that the protective effects of agmatine on Müller cells were inhibited by N-methyl-D-aspartic acid (NMDA). The results of the present study suggested that agmatine treatment protects Müller cells from high-concentration glucose-induced cell damage. The underlying mechanisms may relate to the anti-inflammatory and antiapoptotic effects of agmatine, as well as to the inhibition of the MAPK pathway, via NMDA receptor suppression. Agmatine may be of use in the development of novel therapeutic approaches for patients with diabetic retinopathy.

Background Poly(adenosine diphosphate-ribose) polymerase 1 (PARP-1) plays an important role in the pathogenesis of diabetic retinopathy (DR),and Notch1 signal pathway is one of the important signal transduction pathways in the organism which may antagonize retinal vascular diseases.However,if Notch1 signaling pathway is involved in pathogenesis of DR has not been confirmed yet.Objective This study was to investigate the expressions of Notch1,Dll4,PARP-1,Akt,nuclear factor-κB (NF-κB) and caspase-3 in the retina of diabetic mouse model and retinal vascular endothelial cells (RVECs) under the high glucose.Methods The expressions of Notch1,Dll4,PARP-1,Akt,NF-κB and caspase-3 in the retina of diabetic mouse models were investigated using immunochemistry and Western blot method after the diabetic mouse models were established.And these proteins were detected in retinal RVECs under the high glucose by Western blot.Results The expressions of Notch1,Dll4 and phosphorylated Akt (p-Akt) in retinas reduced significantly and simultaneously companied with increases of PARP-1 and caspase-3 in diabetic mice compared with normal mice (all at P＜0.05).However,no obvious change was found in the expression of NF-κB (P＞0.05).Expressions of Notch1 and p-Akt in RVECs increased with the increase of glucose concentration,but expressions of cleaved PARP-1 and caspase-3 decreased,especially in the 30 mmol/L group,showing significant differences in comparison with the normal control group (all at P＜0.05).But no altering of NF-κB expression was seen in the mice with diabetes mellitus.Conclusions The expressions of cleaved-PARP-1 and caspase-3 in the retinas is up-regulated,but the expressions of Notch1 and p-Akt are down-regulated in diabetic mice.

目的 观察缺氧、氧化应激对体外培养的视网膜色素上皮(retinal pigment epithelium,RPE)细胞增殖的影响及GRP78表达的变化.方法 体外培养人RPE细胞,采用CoCl2诱发细胞缺氧模型、H2O2诱发氧化应激模型.实验分为量-效关系组:细胞分别经浓度为100 μmol·L-1、200 μmol·L-1、400 μmol·L-1、800 μmol·L-的CoCl2、H2O2作用12h,用MTT法检测细胞增殖情况;时-效关系组:在CoCl2浓度为200 μmol·L-1、H2O2浓度为400 μmol·L-1的基础上,分别选取药物作用8h、12 h、16h3个时间点收集细胞,采用免疫细胞化学染色检测GRP78蛋白的表达情况.结果 量-效关系组:不同浓度CoCl2组RPE细胞活性与空白对照组相比均明显下降(均为P＜0.05);400 μmol·L-1及800 μmol·L-1H2O2组细胞存活率较空白对照组均明显下降(均为P＜0.05).时-效关系组:空白对照组GRP78表达阳性细胞率为(5.4±3.0)％;CoCl2作用8h、12 h、16 hGRP78阳性细胞率分别为(34.2±7.1)％、(54.4±9.1)％、(34.7±6.8)％;H2O2作用8h、12 h、16 h GRP78阳性细胞率分别为(37.8±7.1)％、(57.4±5.1)％、(42.5±3.9)％.经统计学分析,CoCl2、H2O2处理8h后,RPE细胞中GRP78表达均较空白对照组明显升高(均为P＜0.01),作用12h时GRP78的表达量最高,作用16 h时GRP78的表达量又有下降.结论 GRP78可以作为RPE细胞缺氧、氧化应激的生化标志物,GRP78的表达变化与细胞应激状态有一定依赖关系.

目的观察重组人色素上皮衍生因子(hPEDF)基因修饰对巨噬细胞条件培养基诱导的视网膜色素上皮细胞(RPE)增殖的影响。方法首先利用脂质体介导基因导入法制备稳定表达hPEDF RPE细胞,MTT法观察巨噬细胞条件培养基诱导RPE细胞及重组hPEDF基因修饰的RPE细胞的增殖作用。结果应用不同浓度的巨噬细胞条件培养基诱导重组hPEDF基因修饰的RPE细胞及未转染的RPE细胞,相应时间的实验组间比较,OD值有显著性差异(P<0.01)。结论重组hPEDF基因修饰可抑制巨噬细胞条件培养基诱导的RPE细胞增殖。

Simvastatin is a cholesterol-lowering drug which exhibits numerous pleiotropic effects including anticancer activity. Yet, the anti-cancer effects in choroidal melanoma remain poorly characterized. Therefore, in this study, we investigated the effects of simvastatin on OCM-1 cells growth, apoptosis and cycle. Simvastatin showed an inhibitory effects on OCM-1 cells viability in dose-dependent (2-10 mu M) and time-dependent (24-72 h) manner. Further study suggested that simvastatin-induced inhibition OCM-1 cells proliferation was associated with G1 phase arrest, decreased protein and mRNA expression of proliferation marker cyclin D1, cyclin E, cyclin dependent kinase (CDK)2 and increased expression of CDK inhibitory protein P21. In addition, simvastatin resulted in an increase in levels of reactive oxygen species (ROS) in OCM-1 cells and simvastatin significantly triggered apoptosis in OCM-1 cells, which was characterized by increased chromatin condensation, activation of caspase-9 and cleaved-caspase-3, increased expression mitochondrion-related apoptosis protein of P53, Bax and decreased expression of Bcl2 and iASPP. Collectively, our study demonstrated that simvastatin can efficiently inhibit proliferation and induce apoptosis in OCM-1 cells. (C) 2013 Elsevier Ltd. All rights reserved.

Objective To investigate the expression and function of intravitreal injection of pigment epitheliumderived factor(PEDF)gene eukaryotic expression vector in proliferative vitreoretinopath(PVR)model eye,and to clarify the inhibitory effect of PEDF gene transfection on PVR induced by macrophages in rats.Methods The pcDNA3-PEDF with cationic liposome was injected into vitreous of 36SD rats(the right eye of each rat as the experimental eye and the left eye as the negative control eye).The rats were divided into PVR control group(injecting macrophages in the vitreous cavity to induce PVR),saline control group(injecting macrophages 3dafter intravitreal injection of saline)and transfection group(injecting macrophages 3dafter intravitreal injection of liposome and pcDNA3-PEDF),and there were 12rats in each group.Then the ophthalmoscope and fundus pre-set lens were used to observe the vitreous and retinal conditions of the rats in each group;the stuctures of the vitreous and retinal layers and the cell proliferation of the specimens of the rats in various groups were detected by HE staining under light microscope.Results The ophthalmoscope and fundus pre-set lens results showed that after intravitreal injection of macrophages,there was more significant vitreous proliferation in PVR control group and saline control group compared with transfection group 1week after injection,there was vitreous proliferation in PVR control group and saline control group.2-3weeks after injection,the proliferation was gradually increased and retina began to detach in PVR control group and saline control group.4 weeks after injection,there was significant proliferation in all model eyes,and traction retinal detachment occurred in PVR control group and saline control group.Only one eye of the rats in transfection group showed vitreous traction and shallow retinal detachment,and the other eyes were normal.The HE staining and light microscope results showed that the structurs of retina cells of most eyes were clear,only one eye occured shallow retinal detachment with epiretinal membrane proliferation and small amount of inflammatory cell infiltration in transfection group.There was inflammatory response with the progression of the disease,accumulation of inflammatory cells,proliferation of fibroblasts,and formation of eyeball atrophy in PVR control group and saline control group.Conclusion PEDF transfected by intravitreal injection in rats' eyes can significantly inhibit the formation and development of PVR induced by macrophages.

<正>1病历摘要患者,男性,70岁,右眼渐进性视物模糊1年,加重1个月,自诉晨起时视力较差,下午视力有所好转,高血压病史4年,糖尿病病史1年。眼科检查:视力:右:指数/40 cm,矫正不应;左:0.8,矫正不应。眼压:右:16 mmHg,左:17 mmHg。右眼:结膜无充血,角膜光滑透明,前房正常深浅,房水清

Aging is a common feature of biological changes in the eye,especially in the Bruch membrane.It is recognized that Bruch membrane changes with the increase of age.Aging may be an important event to contribute to the pathogenesis of macular diseases,such as age-related macular degeneration (AMD).Therefore,this review summarizes the latest concepts about the changes of anatomic structure,physiological function,lipids deposition on Bruch membrane with age,these factors may play an important role in the macular diseases.Further study on these factors will offer new ideas about the etiology,pathology,diagnose and treatment of macular diseases.

患者男,45岁.因间歇性头痛3天后双眼视力障碍1天于2011年11月9日就诊于我院.患者8年前左眼被铁屑崩伤后行球内异物取出、白内障摘除、人工晶状体植入术.2011年9月21日左眼因人工晶状体脱位行人工晶体调位术、前部玻璃体切割术.查体:视力:0D:0.4(矫正不应)OS:0.5(矫正不应).眼压:右:16mm Hg,左:32 mm Hg.双眼红绿色觉:(+),结膜无充血,角膜光滑透明,前房正常深浅,房水清,虹膜纹理清,瞳孔正圆,直径约3.00mm,右眼晶状体前囊可见片状混浊,玻璃体透明.左眼人工晶状体位正,人工晶状体前表面可见点状棕色色素沉着,玻璃体混浊.眼底:视乳头边界欠清,色淡红,A/V=1∶2,后极部视网膜及黄斑区水肿,黄斑区鼻侧视网膜呈放射状皱褶,黄斑中心凹反光(-).初步诊断:交感性眼炎?Vogt-小柳原田病(双)?继发性青光眼(左)并发性白内障(右)人工晶状体眼(左)。

Driver inattention (fatigue and distraction) monitoring method has become a focus of Driver Assistance Systems (DAS). A new algorithm based on machine vision is proposed which includes driver's face location based on gray variance, and segmentation of separate areas of facial organs using the projection curve pole position. The eye contour state can be obtained through OTSU calculation and contour extraction in each separate region. Calculation method of driver's facial rotation angle is presented based on the analysis of driver's head rotation. Different monitoring methods of driver state were used according to different head rotation angles. PERCLOS is used to monitor the fatigue state when the driver faces forward, while facial orientation is used to monitor the distraction state when the driver faces other directions. Results show that the processing algorithm cost 32ms in average, so it has good real-time performance and accuracy rate.

患者男,55岁.因双眼先后视力下降、眼痛伴头痛1个半月入我院,自诉发病前曾有感冒样症状.右眼曾在当地医院行抗青光眼手术治疗,术后症状无改善.入院查体:视力:右眼:手动/20 cm,左眼:0.1(+3.00=0.3);眼压:右眼:12 mm Hg,左眼:19 mm Hg.右眼角膜光滑透明,上方虹膜根切口可见,瞳孔直径约7 mm、欠圆,对光反射(-),晶状体混浊,玻璃体混浊.眼底模糊可见:视乳头充血、水肿,附近视网膜水肿,下方渗出性视网膜脱离,黄斑中心凹反射(-),左眼角膜后KP,瞳孔直径约4.0 mm、欠圆,对光反射(-),晶状体混浊,玻璃体混浊.眼底模糊可见:视盘境界欠清,下方渗出性视网膜脱离,黄斑中心凹反射(-).B超:示双眼玻璃体混浊,渗出性视网膜脱离(图1).

In this study, rabbit models of optic nerve injury were reproduced by the clamp method. After modeling, rabbit models were given one injection of 50 ng recombinant human ciliary neurotrophic factor into the vitreous body and/or intragastric injection of 4 g/kg compound light granules containing Radix Angelicae Sinensis and Raidix Paeoniae Alba at 4 days after modeling, once per day for 30 consecutive days. After administration, the animals were sacrificed and the intraorbital optic nerve was harvested. Hematoxylin-eosin staining revealed that the injured optic nerve was thinner and optic nerve fibers were irregular. After treatment with recombinant human ciliary neurotrophic factor, the arrangement of optic nerve fibers was disordered but they were not markedly thinner. After treatment with compound light granules, the arrangement of optic nerve fibers was slightly disordered and their structure was intact. After combined treatment with recombinant human ciliary neurotrophic factor and compound light granules, the arrangement of optic nerve fibers was slightly disordered and the degree of injury was less than after either treatment alone. Results of tensile mechanical testing of the optic nerve showed that the tensile elastic limit strain, elastic limit stress, maximum stress and maximum strain of the injured optic nerve were significantly lower than the normal optic nerve. After treatment with recombinant human ciliary neurotrophic factor and/or compound light granules, the tensile elastic limit strain, elastic limit stress, maximum stress and maximum strain of the injured optic nerve were significantly increased, especially after the combined treatment. These experimental findings indicate that compound light granules and ciliary neurotrophic factor can alleviate optic nerve injury at the histological and biochemical levels, and the combined treatment is more effective than either treatment alone.