Altered microRNA expression profiles in retinas with diabetic retinopathy

Rats with streptozotocin (STZ)-induced diabetes were studied in order to identify abnormal microRNA (miRNA) expression profiles in diabetic retinopathy (DR) and to ascertain miRNAs associated with DR. Histopathologically, we observed characteristic features of DR in rats at 10 weeks after STZ injection. Investigation of miRNA expression profiles in the retinas of control and diabetic rats using miRNA microarrays revealed that many miRNAs were abnormally expressed in DR. On the basis of their fold changes and probability values, a total of 37 miRNAs were selected for further validation by real-time PCR analysis. The results showed that 11 miRNAs were significantly upregulated and 6 miRNAs were notably downregulated in DR. Furthermore, these changes in retinal miRNA expression levels paralleled the course of DR. Levels of miR-182, miR-96, miR-183, miR-211, miR-204, and miR-124 were significantly increased during the progress of DR, whereas miR-10b, miR-10a, miR-219-2-3p, miR-144, miR-338, and miR-199a-3p were significantly decreased. Our data indicate that the aberrant miRNA expression profiles in DR are associated with the development of DR. Modulation of retinal miRNA expression levels may provide a potential therapeutic strategy for DRs.     

糖尿病患者视网膜神经纤维层OCT测量的形态学观察

背景 传统的观点认为,糖尿病视网膜病变（DR）的主要病理机制是视网膜的微血管病变,但近年来发现糖尿病患者的视功能改变发生于DR的微血管病变发生之前,传统的观点无法解释糖尿病患者在出现可辨认的血管改变之前已有视功能改变的现象. 目的 应用频域光学相干断层扫描（OCT）观察无视网膜血管改变的糖尿病患者、非增生性糖尿病视网膜病变（NPDR）患者视网膜神经纤维层（RNFL）厚度的改变,分析糖尿病患者RNFL厚度与DR的关系. 方法 收集2012年10月至2013年9月于山西省眼科医院就诊且确诊的2型糖尿病患者56例60眼,按DR国际临床分型标准分为非糖尿病视网膜病变（NDR）组26例30眼和NPDR组（轻中度NPDR）30例30眼,同期纳入年龄和性别匹配的健康志愿者30人30眼作为对照.应用OCT对受试者行RNFL厚度测量,包括视盘周围及中心凹周围视网膜360°平均RNFL厚度及鼻上、颞上、颞侧、颞下、鼻下、鼻侧区域的RNFL厚度,比较各组患者RNFL厚度的变化. 结果 NDR组、NPDR组和正常对照组受试者视盘周围平均RNFL厚度分别为（97.46±8.65）、（100.69±16.35）和（109.22±8.69）μm,其中NDR组、NPDR组受试者视盘周围平均RNFL厚度值均低于正常对照组,差异均有统计学意义（P=0.001、0.006）;NDR组视盘周围各象限RNFL厚度均较正常对照组变薄,颞上、颞侧、颞下和鼻下象限RNFL厚度的差异均有统计学意义（P=0.001、0.001、0.001、0.010）;NPDR组视盘周围各象限RNFL厚度均较正常对照组变薄,其中颞上、鼻侧区域RNFL厚度的差异均有统计学意义（P=0.001、0.046）.NDR组、NPDR组和正常对照组平均黄斑区RNFL厚度分别为（33.47±3.39）、（36.81±3.21）、（38.18±2.16）μm,NDR组、NPDR组受试者黄斑区平均RNFL厚度值均低于正常对照组,其中NDR组与正常对照组比较差异有统计学意义（P=0.001）;NDR组黄斑区各区域RNFL厚度较正常     

定量分析糖尿病视网膜病变患者视盘周围视网膜和脉络膜血流灌注

目的 定量分析不同阶段糖尿病视网膜病变(DR)患者视盘周围视网膜和脉络膜变化。方法 对90例(90眼)患者进行横断面研究。受试者分为3组：对照组(n=30)、无DR(NDR)组(n=30)和非增生型DR(NPDR)组(n=30)。比较不同组别受试者视盘周围视网膜血流灌注(RP)、视网膜血管密度(RVD)、视网膜神经纤维层厚度(RNFL)、脉络膜血管体积(CVV)、脉络膜血管指数(CVI)、脉络膜厚度(CT)。结果 与对照组相比,NDR组患者RP[(13.16±0.98)mm²]、RVD[(61.80±5.29)%]、RNFL[(75.41±11.19)μm]均显著降低(均为P<0.05),NPDR组患者RP[(11.70±1.53)mm²]、RVD[(56.92±5.61)%]、RNFL[(68.77±9.61)μm]和CVI(0.16±0.05)均降低(均为P<0.05)。与NDR组相比,NPDR组患者RP、RVD、RNFL和CVI均降低(均为P<0.05)。结论 NPDR期患者视盘周围视网膜和脉络膜血流已发生变化。扫频O...     

NPDR患者黄斑区视网膜厚度、视网膜神经纤维层厚度与全视野闪光ERG的变化

目的探讨非增生性糖尿病视网膜病变(non-proliferative diabetic retinopathy,NPDR)患者黄斑区视网膜厚度(retinal thickness,RT)、视网膜神经纤维层(retinal nerve fiber layer,RNFL)厚度与全视野闪光视网膜电图(electroretinogram,ERG)的变化及相互关系。方法选择2010年2月至12月于我院就诊的2型糖尿病患者91例(168眼),依据糖尿病视网膜病变(diabetic retinopathy,DR)国际分期标准分为3组:无糖尿病视网膜病变(NDR)组39例(78眼),轻度NPDR组28例(52眼),中重度NPDR组24例(38眼)。另选择同龄正常人30例(60眼)作为正常对照组。通过光学相干断层扫描检测黄斑区中心RT及RN-FL厚度,全视野闪光ERG检测视网膜功能。结果 NDR组、轻度NPDR组、中重度NPDR组患者黄斑区RT分别为(198.01±22.51)μm、(218.00±28.15)μm、(239.90±65.04)μm,均较正常对照组(184.45±18.50)μm增厚,差异均有统计学意义(t=2.312、5.235、7.947,均为P<0.05);随着DR加重,糖尿病各组间黄斑区RT比较,差异均有显著统计学意义(t=3.305、6.300、3.069,均为P<0.01)。正常对照组与NDR组黄斑区RNFL厚度比较,差异无统计学意义(P>0.05);轻度NDPR组及中重度NPDR组与正常对照组相比,黄斑区RNFL厚度变薄,差异均有统计学意义(均为P<0.05)。与正常对照组比较,中重度NPDR组全视野闪光ERG暗适应0.01反应及明适应3.0反应b波潜伏期明显延长(均为P<0.05),暗适应0.01反应b波振幅和暗适应3.0反应a波振幅均下降(均为P<0.05)。结论随着NPDR病情加重,2型糖尿病患者黄斑区中心RT增加,RNFL厚度减小,视网膜功能受损。     

全视网膜光凝术对糖尿病视网膜病变视网膜神经纤维层及黄斑区视网膜的影响

目的　探讨全视网膜光凝术（ｐａｎｒｅｔｉｎａｌｐｈｏｔｏｃｏａｇｕｌａｔｉｏｎ,ＰＲＰ）对糖尿病视网膜病变（ｄｉａｂｅｔｉｃｒｅｔｉｎｏｐａｔｈｙ,ＤＲ）视网膜神经纤维层（ｒｅｔｉｎａｌｎｅｒｖｅｆｉｂｅｒｌａｙｅｒ,ＲＮＦＬ）及黄斑区视网膜的影响。方法　选取２０１０年６月至２０１３年１２月于我院行ＰＲＰ治疗的１２０例（１２０眼）ＤＲ患者,其中非增生性糖尿病视网膜病变（ｎｏｎ-ｐｒｏｌｉｆｅｒａｔｉｖｅｄｉａｂｅｔｉｃｒｅｔｉｎｏｐａｔｈｙ,ＮＰＤＲ）６０例（ＮＰＤＲ组）,增生性糖尿病视网膜病变（ｐｒｏｌｉｆｅｒａｔｉｖｅｄｉａｂｅｔｉｃｒｅｔｉｎｏｐａｔｈｙ,ＰＤＲ）６０例（ＰＤＲ组）,同时选取同期于我院健康体检的６０例（６０眼）正常志愿者作为正常对照组。ＰＲＰ手术前后使用ＯＣＴ横向扫描视盘旁ＲＮＦＬ厚度和黄斑区,将视盘旁ＲＮＦＬ和黄斑区分为上方、鼻侧、下方和颞侧４个象限进行扫描,获取各象限及全周平均视盘旁ＲＮＦＬ厚度及黄斑区视网膜厚度,并对结果进行统计分析。结果　与正常对照组相比,ＮＰＤＲ和ＰＤＲ组ＰＲＰ前视盘上方、下方、鼻侧象限、全周平均ＲＮＦＬ厚度均降低（均为Ｐ＜０．０５）,但颞侧象限ＲＮＦＬ厚度差异均无统计学意义（均为Ｐ＞０．０５）;ＮＰＤＲ组和ＰＤＲ组ＰＲＰ前不同象限及全周平均ＲＮＦＬ厚度相比,差异均无统计学意义（均为Ｐ＞０．０５）。与ＰＲＰ前相比,ＮＰＤＲ、ＰＤＲ组ＰＲＰ后各象限及全周平均ＲＮＦＬ厚度均变薄,但只有上方、下方及全周平均ＲＮＦＬ厚度差异均有统计学意义（均为Ｐ＜０．０５）;与ＮＰＤＲ组相比,ＰＤＲ组上方、下方、鼻侧及全周平均ＲＮＦＬ厚度变薄更明显,差异均有统计学意义（均为Ｐ＜０．０５）。与正常对照组相比,ＮＰＤＲ和ＰＤＲ组ＰＲＰ前各象限及平均黄斑区视网膜厚度均增加,差异均有统计学意义（均为Ｐ＜０．０５）,且ＰＤＲ组较ＮＰＤＲ组增加更为明显,差异均有统计学意义（均为Ｐ＜０.０５）。与ＰＲＰ前相比,ＮＰＤＲ、ＰＤＲ组ＰＲＰ后各象限及平均黄斑区视网膜厚度均增加,差异均有统计学意义（均为Ｐ＜０．０５）,且ＰＤＲ较ＮＰＤＲ组增加更为明显,差异均有统计学意义（均为Ｐ＜０．０５）。结论　ＰＲＰ对ＤＲ患者ＲＮＦＬ有一定损伤,应选择适当的激光能量与曝光时间,最大限度地降低对ＲＮＦＬ的影响。     

Analysis of MicroRNA Expression in Tears of Patients with Herpes Epithelial Keratitis: A Preliminary Study

PurposeHerpes epithelial keratitis (HEK) is the most common form of herpes simplex virus (HSV) eye involvement, and understanding the molecular mechanisms underlying HEK is important. We investigated the expression of microRNAs (miRNAs) in the tears of patients with HEK.MethodsTear samples from eight patients with HEK and seven age-matched controls were evaluated. Clinical ophthalmologic evaluation was performed, and an anterior segment photograph was obtained after fluorescence staining. Dendritic or geographic ulcer areas were measured using ImageJ software. The expression of 43 different miRNAs in tears was measured using real-time polymerase chain reaction and compared between patients with HEK and controls. Differences in miRNA expression between the dendritic and geographic ulcer groups and correlations involving miRNA expression and ulcer area were evaluated.ResultsOf the 43 miRNAs, 23 were upregulated in patients with HEK compared to normal controls. MiR-15b-5p, miR-16-5p, miR-20b-5p, miR-21-5p, miR-23b-3p, miR-25-3p, miR-29a-3p, miR-30a-3p, miR-30d-5p, miR-92a-3p, miR-124-3p, miR-127-3p, miR-132-3p, miR-142-3p, miR-145-5p, miR-146a-5p, miR-146b-5p, miR-155-5p, miR-182-5p, miR-183-5p, miR-221-3p, miR-223-3p, and miR-338-5p were significantly upregulated in patients with HEK. MiR-29a-3p exhibited significant differences between the dendritic and geographic ulcer groups. All 23 miRNAs with significant differences between patients with HEK and the control group were not significantly correlated with ulcer area.ConclusionsTwenty-three miRNAs were significantly upregulated in the tears of patients with HEK, and the expression of miRNAs may play important roles in herpes infection in relation to host immunity.     

Anemia and Diabetic Kidney Disease Had Joint Effect on Diabetic Retinopathy Among Patients With Type 2 Diabetes

PurposeWhether the association between diabetic kidney disease (DKD) and diabetic retinopathy (DR) in patients with type 2 diabetes mellitus (T2DM) is leveraged by anemia remains unclear. This study is to evaluate the joint effect of DKD and anemia on DR.MethodsData were collected from electronic medical records of 1389 patients with T2DM in the Yiwu Central Hospital of Zhejiang Province from 2018 to 2019. Based on retinal examination findings, patients were classified as without diabetic retinopathy (non-DR), non-proliferative diabetic retinopathy (NPDR), and proliferative diabetic retinopathy (PDR). Odds ratio (OR) from multinomial logistic regression models adjusting for potential risk factors of DR were used to evaluate associations of DKD, renal function measures, and anemia with risk of NPDR and PDR. Path analysis was performed to help understand the association of DKD and hemoglobin (Hb) with DR.ResultsThe study included 901 patients with non-DR, 367 patients with NPDR and 121 patients with PDR. Both high DKD risk and abnormal renal function measures were significantly associated with PDR. Anemia was associated with increased risk of NPDR (OR = 1.75, 95% confidence interval [CI] = 1.18–2.58) and PDR (OR = 3.71, 95% CI = 2.23–6.18). DKD severity and anemia had joint effect on NPDR (OR = 2.29, 95% CI = 1.32–3.96) and PDR (OR = 11.31, 95% CI = 5.95–21.51). These associations were supported by path analysis.ConclusionsDKD severity, abnormal estimated glomerular filtration rate (eGFR), and urinary albumin/creatinine ratio (UACR) were associated with increased risk of DR in patients with T2DM, and anemia had joint effect on these associations. Improving Hb level may decrease the risk of DR in patients with T2DM.     

早期糖尿病视网膜病变患者黄斑神经节细胞层厚度及视盘参数的变化

目的:使用Cirrus HD-OCT比较早期糖尿病视网膜病变患者和健康对照者之间的视盘参数、视盘周围视网膜神经纤维层(pRNFL)厚度和黄斑神经节细胞层(mGCL)厚度改变。方法:横断面比较研究。选取45例无糖尿病视网膜病变(NDR)、52例轻度非增殖期糖尿病视网膜病变(NPDR)及55例中度NPDR的2型糖尿病患者和64例年龄匹配的健康对照者纳入本研究。采集患者的空腹血糖、糖尿病病程、糖化血红蛋白及既往史。通过Cirrus HD-OCT测量视盘参数(即双眼RNFL厚度对称性百分比、盘沿面积、视盘面积、杯盘比、杯容积)、pRNFL厚度和mGCL厚度。组间比较采用单因素方差分析。结果:与健康对照组比较,NDR、轻度NPDR、中度NPDR组双眼RNFL厚度对称性百分比、盘沿面积明显降低,平均C/D、垂直C/D明显增加(均P<0.05)。与健康对照组比较,NDR、轻度NPDR、中度NPDR组患眼视盘周围各象限(上方、颞侧、下方、鼻侧)及平均RNFL厚度和黄斑(平均、最小、上方、颞上方、颞下方、下方、鼻上方、鼻下方)GCL厚度显著变薄(均P<0.05)。结论:与健康对照者相比,早期...     

糖尿病视网膜病变患者血清微核糖核酸表达谱的初步研究

目的本研究旨在探讨血清微核糖核酸(miRNA)成为糖尿病视网膜病变(DR)生物标志物的可能性。 方法队列研究。2014年11月至2015年6月,招募中国医科大学附属盛京医院眼科和内分泌科患者69例作为研究对象。其中,单纯糖尿病患者30例作为单纯糖尿病组,DR患者39例作为DR组。两组采用数字表法各随机选取3例患者,应用miRNA芯片技术检测血清miRNA的表达水平;使用分层聚类分析,获得DR患者的血清miRNA表达谱;利用生物信息学方法,分析特异性表达的血清miRNA的靶基因及其相关信号通路。采用实时荧光定量聚合酶链式反应(qRT-PCR)技术,检测两组患者血清中特异性miRNA的表达水平。采用受试者工作特征曲线(ROC曲线),评价血清中特异性miRNA对DR的诊断效能。 结果miRNA芯片技术检测表明,在3100种成熟的miRNA中,共筛选出19种差异miRNA。其中,13种表达上调;6种表达下调。结果表明,在DR患者血清中存在特异性表达的miRNA。qRT-PCR检测表明,DR组miR-19b、miR-221和miR-18b表达均上调,差异有统计学意义(U＝256.027,125.515,254.017;P<0.05),并且与miRNA芯片筛选的结果相符。受试者工作特征曲线结果表明,miR-19b(曲线下面积＝0.78,95%CI＝0.66~0.90)、miR-221(曲线下面积＝0.89,95%CI＝0.81~0.97)和miR-18b(曲线下面积＝0.78,95%CI＝0.67~0.90)对DR均有较高的诊断效能,其中miR-221诊断效能最高。 结论DR患者血清中存在特异性miRNA表达谱,这些差异性表达的miRNA可能作为调控因子,通过调节靶基因调控DR的发生和发展,其中miR-221最有望成为DR的生物标志物。     

Peripapillary vessel density and retinal nerve fiber layer thickness changes in early diabetes retinopathy

AIM: To perform a quantitative analysis of the peripapillary vessel density (VD) and retinal nerve fiber layer (RNFL) thickness changes in patients with early stage of diabetic retinopathy (DR). METHODS: In this case-control study, swept-source optical coherence tomography angiography (SS-OCTA) imaging was used to examine diabetic and age-matched healthy subjects. The optic disc HD 6×6 mm2 blood flow imaging scan mode was selected. Automatic software was used to measure the peripapillary VD, capillary vessel density (CVD), and RNLF in an optic nerve head (ONH) filed based on the Garway-Heath map. In addition, the correlation between peripapillary VD, CVD, and RNFL was further investigated. RESULTS: The cohort consisted of 32 healthy individuals and 72 patients with diabetes (34 eyes with no DR and 38 eyes with mild-moderate NPDR). Peripapillary VD decreased in the mild-moderate NPDR group compared to the control group in most regions (P<0.05). Peripapillary CVD and RNFL thickness were significantly lower in the mild-moderate NPDR group in the superior temporal (ST) quadrants (P=0.018, P=0.030). In the correlation analysis of each region, the RNFL thickness in the NS region was positively correlated with the peripapillary VD and CVD (r=0.233, P=0.05; r=0.288, P=0.015). In the TI region, the RNFL thickness was positively correlated with the peripapillary CVD (r=0.237, P=0.047). CONCLUSION: The measurement based on the ONH topographic map may be helpful in detecting functional and structural impairments in DR. The peripapillary VD, CVD and RNFL decrease in early DR, and the RNFL thickness altered in association with the CVD or/and VD in some regions.     

miR-132在糖尿病视网膜病变患者血浆中的表达及其临床意义

目的：分析miR-132在糖尿病视网膜病变(DR)患者血浆中的表达及与DR的关系。

方法：前瞻性研究,选取2015-07/10于我院确诊的55例糖尿病患者,按DR国际临床分期标准将患者分为5组。背景期：无明显视网膜病变组13例(A组); 非增殖期(NPDR)33例,包括：轻度NPDR组10例(B组),中度NPDR组11例(C组),重度NPDR组12例(D组); 增殖期(PDR)9例(E组)。另选取我院体检健康者12例作为健康对照组(F组)。应用实时荧光定量PCR(qRT-PCR)技术检测不同分期DR患者外周血浆中miR-132的相对表达量,并比较各组间差异。

结果：除无明显视网膜病变组外,其余各组DR患者与健康对照组相比,血浆中miR-132的表达水平均明显下降(P<0.05); 非增殖期组间、非增殖期与增殖期组间比较,miR-132表达量无差异(P>0.05)。

结论：miR-132在NPDR和PDR患者血浆内低表达,可能成为DR治疗的生物标志物。     

Association between risk factors and retinal nerve fiber layer loss in early stages of diabetic retinopathy

AIM:To investigate the changes of retinal nerve fiber layer(RNFL)among normal individuals,diabetic patients without diabetic retinopathy(NDR)and non-proliferative diabetic retinopathy(NPDR),and explore the possible risk factors of early diabetic retinopathy(DR).METHODS:In this cross-sectional study,107 participants were divided in three groups.Totally 31 normal individuals(control group),40 diabetic patients without DR(NDR group)and 36 patients with NPDR(NPDR group)were included.Optical coherence tomography(OCT)was used to detect RNFL thickness and other optic disc parameters among different groups.The potential association between RNFL loss and systemic risk factors were assessed for DR,including diabetes duration,body mass index(BMI),hemoglobin A1 c(Hb A1 c),serum lipids,and blood pressure.RESULTS:The average and each quadrant RNFL thickness were thinner in NPDR group compared to control group of the right(P=0.00,P=0.01,P=0.01,P=0.02,P=0.04)and left eyes(P=0.00,P=0.00,P=0.00,P=0.03,P=0.04).The average,superior and inferior RNFL thickness were thinner in NDR group compared to the NPDR group of the right(P=0.00,P=0.02,P=0.03)and left eyes(P=0.00,P=0.00,P=0.01).Diabetic duration was negatively correlated with the superior,inferior,and average RNFL thickness of the right(r=-0.385,P=0.001;r=-0.366,P=0.001;r=-0.503,P=0.000)and left eyes(r=-0.271,P=0.018;r=0.278,P=0.015;r=-0.260,P=0.023).Hb A1 c was negatively correlated with the superior,inferior,and average RNFL thickness of the right(r=-0.316 P=0.005;r=-0.414,P=0.000;r=-0.418,P=0.000)and left eyes(r=-0.367,P=0.001;r=-0.250,P=0.030;r=-0.393,P=0.000).Systolic pressure was negatively correlated with the inferior and average RNFL thickness of the right eye(r=-0.402,P=0.000;r=-0.371,P=0.001)and was negatively correlated with the superior and average RNFL thickness of the left eye(r=-0.264,P=0.021;r=-0.233,P=0.043).CONCLUSION:RNFL loss,especially in the superior and inferior quadrants,may be the earliest structural change of the retina in diabetic patients,and is also associated with diabetic duration,Hb A1 c,and systolic pressure.     

Retrobulbar administration of purified anti-nerve growth factor in developing rats induces structural and biochemical changes in the retina and cornea

AIM:To develop an experimental model of endogenous nerve growth factor(NGF)deprivation by retrobulbar administration of purified neutralizing anti-NGF antibodies in young Sprague-Dawley rats and provide further information on NGF expression in the retina and cornea.METHODS:Sixty old pathogen-free Sprague Dawley rats(p14,post-natal days)were treated with repeated retrobulbar injections of neutralizing anti-NGF(2μL,100μg/m L,every 3 d).After 2 wk(p28),retinal and corneal tissues were investigated for morphological,biochemical,and molecular expression of trkANGFR by using Western blotting or immunofluorescence.Rhodopsin as well as protein profile expression were also investigated.RESULTS:Chronic retrobulbar neutralizing anti-NGF antibodies changed the distribution of trkANGFR immunoreactivity at retinal level,while no changes were detected for global trkANGFR protein expression.By contrary,the treatment resulted in the increase of corneal trkANGFR expression.Retinal tissues showed a decreased rhodopsin expression as well as reduced number of both rhodopsin expressing and total retinal cells,as observed after single cell extraction.A decreased expression of ICAM-1,IL-17 and IL-13 as well as an increased expression of IL-21 typified retinal extracts.No significant changes were observed for corneal tissues.CONCLUSION:The reduced availability of endogenous NGF,as produced by chronic retrobulbar anti-NGF administration,produce a quick response from retinal tissues,with respect to corneal ones,suggesting the presence of early compensatory mechanisms to protect retinal networking.     

不同病理类型的葡萄膜黑色素瘤中微小RNA差异表达谱分析

背景 目前研究证实微小RNA(miRNA)参与大多数人类肿瘤疾病的发生和发展,其作用类似于抑癌基因或癌基因.葡萄膜黑色素瘤(UM)是成人常见的眼部恶性肿瘤,其发生和转移机制仍未完全阐明.探讨UM组织中miRNA的差异表达情况有望为UM的靶向治疗提供依据. 目的 筛选不同病理类型的UM组织中特异性miRNA表达谱. 方法 收集于2013年3月至2015年10月在北京同仁医院手术局部切除并经常规组织病理学和免疫组织化学检测证实为梭型细胞型UM的标本4例和上皮细胞型UM标本4例,采用miRNA芯片分别检测2种UM组织中miRNA的表达,收集同期死于非肿瘤疾病的8个供体眼的正常葡萄膜组织作为对照,利用组间差异倍数筛选出差异≥2倍差异表达的miRNA;用在线软件预测差异表达miRNA的靶基因,采用生物信息学方法分析靶基因参与的信号功能通路.采用实时定量PCR法验证芯片检测结果.结果 收集的梭形细胞型和上皮细胞型UM标本经组织病理学检查均得到确诊,免疫组织化学检测梭形细胞型及上皮细胞型UM组织中HMB45、黑色素-A和S-100均呈阳性反应.与正常葡萄膜组织比较,在梭形细胞型UM组织中差异表达的miRNA有109个,其中29个上调,80个下调,上调的miRNA包括miR-146a-5p、miR-25-3p和miR-29b-1-5p,下调的miRNA包括miR-126-5 p、miR-183-5p和miR-96-5p;上皮细胞型UM中差异表达的miRNA有50个,其中23个上调,27个下调,上调的miRNA包括miR-155-5p、miR-210和miR-378 a-5p;下调的miRNA包括miR-199a-5p、miR-143-3p和miR-143-5p.在梭形细胞型和上皮细胞型UM组织中共同上调的miRNA为miR-132-3p、miR-21-5p、miR-34a-5p和miR-34b-5p,共同下调的miRNA为miR-125b-2-3p、miR-126-3p、miR-199a-3p和miR-214-3p.梭形细胞型和上皮细胞型UM组织中差异表达的miRNA所预测的靶基因分别参与癌症通路、丝裂原活化蛋白激酶(MAPK)信号通路、Wnt信号通路、细胞间黏附、胞吞作用、前列腺癌通路、结直肠癌通路和细胞黏附通路.结论 与正常葡萄膜组织相比,梭形细胞型UM和上皮细胞型UM组织中存在多种miRNA的差异表达,梭形细胞型UM和上皮细胞型UM组织之间也存在明显的miRNA差异表达,这些差异表达的miRNA可通过不同的信号转导通路参与调控UM的生物学行为.     

Assessment of retinal neurodegeneration with spectral-domain optical coherence tomography: a systematic review and meta-analysis

ObjectivesTo comprehensively assess diabetic retinopathy neurodegeneration (DRN) as quantified by retinal neuronal and axonal layers measured with spectral-domain optical coherence tomography (SD-OCT) in subjects with diabetes mellitus (DM).MethodsArticles on the topic of examining macular ganglion cell-inner plexiform layer (m-GCIPL), macular retinal nerve fibre layer (m-RNFL), macular ganglion cell complex (m-GCC), and peripapillary RNFL (p-RNFL) measured with SD-OCT in DM subjects without DR (NDR) or with non-proliferative DR (NPDR) were searched in PubMed and Embase up to November 31, 2019. Standardized mean difference (SMD) as effect size were pooled using random-effects model.ResultsThirty-six studies searched from online databases and the CUHK DM cohort were included in the meta-analysis. In the comparison between NDR and control, macular measures including mean m-GCIPL (SMD = −0.26, p = 0.003), m-RNFL (SMD = −0.26, p = 0.046), and m-GCC (SMD = −0.28; p = 0.009) were significantly thinner in the NDR group. In the comparison between NPDR and NDR, only mean p-RNFL was significantly thinner in the NPDR group (SMD = −0.27; p = 0.03), but not other macular measures.ConclusionsThinning of retinal neuronal and axonal layers at macula as measured by SD-OCT are presented in eyes with NDR, supporting DRN may be the early pathogenesis in the DM patients without the presence of clinical signs of DR. In the future, these SD-OCT measures may be used as surrogates of DRN to stratify DM patients with a high risk of DR, and may be used as a therapeutic target if neuroprotection treatment for DR is available.Subject terms: Retinal diseases, Eye abnormalities     

miR-26a/PTEN在糖尿病小鼠视网膜神经变性中的作用及其机制

目的　探讨 microRNA-26a-5p (miR-26a)/PTEN在早期糖尿病小鼠视网膜神经变性（DRN）中的作用及其机制。方法　66只C57BL/6J小鼠随机分为实验组和对照组,实验组小鼠腹腔注射链脲佐菌素建立糖尿病动物模型。实验组小鼠造模成功后随机分为糖尿病组和miR-26a组,miR-26a组小鼠玻璃体内注射miR-26a mimic上调视网膜组织中miR-26a的表达。玻璃体内注射2周后所有小鼠用100 g·L^-1水合氯醛腹腔注射麻醉并处死,立即摘除眼球。HE染色及透射电镜分别观察各组小鼠视网膜形态学及超微结构变化,免疫荧光半定量检测并定位PTEN、胶质细胞原纤维酸性蛋白（GFAP）在小鼠视网膜各层的表达,qRT-PCR测量各组小鼠视网膜中miR-26a、PTEN和GFAP的mRNA表达。结果　糖尿病组小鼠视网膜神经上皮层厚度较对照组变薄,视网膜神经节细胞(RGC)数量减少,差异均有统计学意义（均为 P<0.05）;视网膜组织中miR-26a mRNA表达下降（P<0.05）,GFAP及PTEN的mRNA表达较对照组小鼠均明显升高（均为 P<0.05）。与糖尿病组小鼠相比,miR-26a组小鼠视网膜全层厚度明显增厚（均为 P<0.05）,RGC丢失减少,视网膜组织中miR-26a mRNA表达升高,而GFAP及PTEN的mRNA表达均降低,差异均有统计学意义（均为 P<0.05）。结论　miR-26a可能通过下调视网膜组织中PTEN的表达减轻糖尿病小鼠的DRN。     

Assessment of optical coherence tomography angiography and multifocal electroretinography in eyes with and without nonproliferative diabetic retinopathy

Purpose:To examine (i) the retinal structure and function using optical coherence tomography angiography (OCTA) and multifocal electroretinography (mfERG), respectively, in eyes with and without nonproliferative diabetic retinopathy (NPDR), (ii) and their interrelationship between retinal structure (OCTA) and function (mfERG) in the two groups independently.Methods:This was a prospective observational study. One hundred twenty-one eligible participants with type 2 diabetes with No DR (n = 89), or with mild or moderate NPDR (n = 32) underwent ophthalmic examination, ultrawide field-view fundus photography, OCTA, and mfERG. Group differences were assessed using a Mann–Whitney U test. Correlations were assessed using Spearman''s rho.Results:There were no significant differences in OCTA measures between the two groups. The mfERG P1 implicit times (rings 1–6) were significantly delayed and P1 response densities in rings 5 and 6 were significantly lower in participants with NPDR compared to those with No DR. In those with No DR, P1 implicit times in almost all rings were delayed in relation to lower vessel density and perfusion (maximum variance noted was 13%). In individuals with NPDR, the P1 response density in rings 2 and 3 showed a positive nonsignificant correlation with macular perfusion.Conclusion:In those with diabetes with No DR, retinal neuronal function is influenced by lower macular vessel density and perfusion. The retinal neuronal function is abnormal in individuals with NPDR compared to those with No DR and is not correlated with OCT angiometric measures, suggesting the likelihood of a different retinal structural correlate.     

外周血液中miRNA表达与老年性黄斑变性相关性研究

目的　检测微小核糖核酸（microribonucleicacids，miRNA）在老年性黄斑变性（age-related macular degeneration，AMD）患者中的表达，并探讨miRNA的表达量与AMD病程之间的关系。方法　选取2014年1月至2016年11月于同济大学附属第十人民医院眼科门诊就诊的AMD患者6例为试验组，并选取同期6名正常人为对照组，通过基因芯片技术检测两组血液中miRNA的表达量。扩大样本的病例对照研究中共纳入126例AMD患者和140名正常人，检测其血液样本中miRNA的表达，比较两组人群间miRNA的表达量差异。结果　通过基因芯片技术，在试验组与对照组间共检测出216个miRNA存在表达差异（均为P<0.05），与对照组相比，试验组中111个miRNA表达量上升，105个miRNA表达量下降，差异均有统计学意义（均为P<0.05）。扩大样本的病例对照研究结果表明，在AMD患者中，miR-27a-3p、miR-29b-3p、miR-195-5p的表达量显著上升，同时，湿性AMD患者血液中miR-27a-3p的表达量高于干性AMD患者，差异均有统计学意义（均为P<0.05）。结论　AMD患者外周血中miRNA表达量水平有明显变化，miR-27a-3p、miR-29b-3p、miR-195-5p可能成为AMD血清学诊断和预后的标志物。