目的：研究玻璃膜疣主要成分胆固醇对人视网膜色素上皮细胞ARPE-19中金属硫蛋白表达的影响。方法：体外培养ARPE-19细胞，将细胞分为对照组和胆固醇处理组(2.5 mg/mL)，取样时间为0，6，12，24，48，72h。通过实时定量PCR检测hMT1a，hMT2a和hMT3在转录水平的表达，用蛋白质印迹法检测总金属硫蛋白的表达。结果：在转录水平上hMT1a，hMT2a和hMT3受到胆固醇影响mRNA表达上调，且hMT3上调倍数最大；总金属硫蛋白的蛋白表达随着胆固醇处理时间延长明显增多。结论：玻璃膜疣主要成分胆固醇可以上调人视网膜色素上皮细胞中金属硫蛋白的表达，提示金属硫蛋白表达可受到玻璃膜疣形成起始阶段的刺激，其检测是否能用于年龄相关性黄斑变性的早期发现及早期诊断还需深入探讨。
Objective: To study the effects of cholesterol, the main component of drusen, on the expression of metallothionein of ARPE-19 cells. Methods: The ARPE-19 cell line was cultured in vitro, and the cells were divided into a control group and a cholesterol treatment group (2.5 mg/mL). The treatment time was 0, 6, 12, 24, 48, 72 hours. Real- time quantitative PCR was used to detect the expression of hMT1a, hMT2a and hMT3 at the mRNA level, and Western blot was used to detect the expression at the protein level. Results: The mRNA expression of hMT1a, hMT2a and hMT3 were up-regulated by cholesterol and the protein expression of total MTs was increased with cholesterol treatment. Conclusion: Cholesterol, the main component of drusen, can up-regulate the expression of metallothionein in human retinal pigment epithelial cells, suggesting that the expression of metallothionine can be stimulated by the initial stage of drusen formation. However, whether its detection can be used for the early detection and early diagnosis of age-related macular degeneration or not still needs to be further explored.
目的：对视网膜光学相干断层扫描图像中不同层和积液区域的分割。方法：提出一种基于深度学习的轻量级的神经网络，参考DRUNet体系、膨胀卷积和残差网络的架构，通过连接不同深度网络处得到的上采样输出，进行多尺度特征融合，使网络能够更好地识别出图像中的边界信息。结果：改进型DRUNet显著提升了视网膜分层的效果，准确率较U-Net提高了1.25%，同时能提前1~2次迭代达到传统U-Net的准确度。结论：本文采用的网络结构提高了对视网膜光学相干断层扫描图像的分割性能，同时降低了网络参数，具有强大的应用潜力。
Objective: To achieve the segmentation of different layers and fluid areas on the optical coherence tomography (OCT) image of the retina. Methods: A lightweight neural network based on deep learning was proposed. The network structure adopted in this study was designed based on the architecture of dilated-residual U-Net. By connecting the upsampling output obtained at different depth networks, multi-scale feature fusion was performed to enable the system to accurately identify the boundaries on the OCT image. Results: Compared with U-Net, this algorithm could achieve the same accuracy with 1–2 epochs less, and the accuracy was also improved by 1.25%. Conclusion: The proposed network improves the segmentation performance of retinal OCT images, and reduces the number of parameters, which demonstrates the network has great application potential.
目的：评价不同操作者使用视网膜自动图像分析(automated retinal image analyzer，ARIA)软件测量糖尿病患者视网膜血管管径和迂曲度的重复性和再现性。方法：采用前瞻性诊断性试验研究设计，由2名熟练操作者应用ARIA软件对49名糖尿病患者以视盘为中心的45°数码眼底彩照进行视网膜血管半自动分析。通过测量距离视盘0.5~1.0个视盘直径内的视网膜动静脉血管参数，各选取6条最粗的动静脉综合计算得到中央动脉管径当量(central retinal artery equivalent，CRAE)、视网膜中央静脉管径当量(central retinal vein equivalent，CRVE)、视网膜动静脉比值(arteriole-to-venule ratio，AVR)、平均视网膜动脉迂曲度(mean retinal arteriolar tortuosity，MRAT)、平均视网膜静脉迂曲度(mean retinal venular tortuosity，MRVT)。在此基础上，评价同一操作者与不同操作者间测量结果的重复性和再现性，并绘制Bland-Altman图进行一致性分析。结果：操作者A两次测量的CRAE、CRVE、AVR差异无统计学意义(t= -0.089、-1.425、1.214，均P>0.05)，重复性良好(ICC≥0.725)且高度正相关(r≥0.721)。操作者A与操作者B对同一受试者2次测量的CRAE、CRVE、AVR差异无统计学意义(t= -1.169、-0.050、0.941，均P>0.05)，再现性良好(ICC≥0.750)且高度正相关( r≥0.697)。同一操作者和不同操作者间的MR AT、MRVT差异无统计学意义(P≥0.05)，Spearman非参数相关性分析重复性和再现性较好(r≥0.571，r≥0.588，均P<0.01)。结论：利用ARIA软件测量糖尿病患者视网膜血管管径及迂曲度具有良好的重复性和再现性。
Objective: To assess the repeatability and reproducibility in measurements of retinal vessel diameter and tortuosity in diabetic patients using the Automated Retinal Image Analyzer (ARIA). Methods: A prospective diagnostic trial was performed; 2 skilled researchers performed a semi-automatic analysis of retinal vessels in digital color fundus photography (45° field of view centered on the optic disc) from 49 diabetic patients using ARIA software. Six largest arterioles and venules coursing through an area of 0.5–1.0 disc diameter surrounding the optic disc were selected. Then the retinal vessel parameters were calculated into the central retinal artery equivalent (CRAE), central retinal vein equivalent (CRVE), arteriole-to-venule ratio (AVR), mean retinal arteriolar tortuosity (MRAT) and mean retinal venular tortuosity (MRVT). To evaluate the repeatability of those values in 2 measurements by the same researcher and the reproducibility between different researchers. Bland-Altman analysis was employed to assess the agreement of the outcomes. Results: There was no significant difference in the CRAE, CRVE, and AVR values in the two-time measurements of researcher A (t= -0.089, -1.425 and 1.214, all P>0.05). It also showed a good repeatability and a highly positive correlation (ICC≥0.725, r≥0.721). There was no significant difference in those measurements between researcher A and B (t= -1.169, -0.050, 0.941, all P>0.05). It also showed a good reproducibility and a highly positive correlation (ICC≥0.750, r≥0.697). Results showed that there was no significant difference in the MRAT and MRVT values both in intraobserver and interobserver measurements (P≥0.05). Besides, Spearman’s nonparametric correlation analysis showed good repeatability and reproducibility (r≥0.571, r≥0.588, all P<0.01). Conclusion: ARIA software yields good repeatability and reproducibility in the measurements of retinal vessel diameter and tortuosity in diabetic patients.
视网膜退行性疾病的种类繁多、患病人口基数大，该病特征为终末期严重的视网膜细胞丢失。视网膜类器官(retinal organoid，RO)可通过3D干细胞体外分化培养技术大量获取，并拥有完整的各亚型视网膜细胞和经典的视网膜分层结构。因此，RO可作为最佳的视网膜退行性疾病建模方法之一，以便于发现潜在致病机制。目前，RO衍生物已被广泛用于视网膜细胞替代治疗的动物实验和临床研究，具体的成效参差不齐，可能的影响因素包括移植细胞数量、移植时间窗、移植工具等。随着RO相关研究的快速发展，视网膜退行性疾病在分子和个体上的诊断和治疗将进一步完善。
Retinal degenerative diseases, characterized by severe retinal cell loss at the end stage, are of various kinds and haunt vast amounts of patients. Retinal organoid (RO) with complete retinal cell subtypes and classic retinal stratification structures can be obtained in large quantities through stem cells in vitro 3D differentiation and culture method. Therefore, RO can serve as one of the best ways for retinal degenerative disease modeling to facilitate the decipherment of underlying pathogenic mechanisms. At present, RO derivatives have been widely used in animal experiments and clinical studies of retinal cell replacement therapy with varying results possibly affected by cell quantity, time window, or tools in terms of transplantation. With the booming progress of RO-related research, the diagnosis and treatment on molecular and individual level for retinal degenerative diseases will be further improved.
目的：建立能驱动GFP在视网膜神经节细胞(retinal ganglion cell，RGC)中特异性表达的小鼠胚胎干细胞系。方法：通过同源重组的方式建立Brn3b-GFP敲入的小鼠胚胎干细胞系(Brn3b-GFP ESC)，利用3D培养将其诱导成视网膜类器官检测GFP表达的细胞特异性，再用流式细胞分选富集GFP阳性RGC，采用玻璃体腔注射的方式将GFP阳性RGC移植到健康小鼠和NMDA损伤模型小鼠眼中探索该细胞的应用价值。结果：Brn3b-GFP ESC经3D视网膜诱导培养后在RGC中特异性表达GFP，将这些GFP阳性RGC移植到两种小鼠中2周后能在所有视网膜内观察到GFP阳性细胞存活，且均能观察到有供体RGC整合到宿主视网膜RGC层。结论：本研究建立了RGC特异的报告基因干细胞系Brn3b-GFP ESC，通过将该细胞系诱导成视网膜类器官进而获得的GFP阳性RGC移植后能够整合进宿主视网膜。该细胞系的建立将为青光眼及相关疾病提供重要的研究手段和工具。
Objective: This study was designed to establish a mouse embryonic stem cell line that can drive GFP expression specifically in retinal ganglion cells (RGCs). Methods: In this study, we established a Brn3b-GFP knock-in embryonic stem cell line (Brn3b-GFP ESC) by homologous recombination. By 3D culture, we induced these cells into retinal organoids to investigate the cell-specificity of GFP expression. GFP-positive RGCs were then enriched by flow cytometry and transplanted by intravitreal injection into the eyes of healthy mice and NMDA injury model mice to explore the feasibility of a potential clinical application. Results: GFP was specifically expressed in RGCs following induction of Brn3b-GFP ESCs into 3D retinal organoids. Two weeks after these GFP-positive RGCs were transplanted into the control and injured mice, GFP-positive cells were observed in all transplanted retinas, and donor RGCs were seen to integrate into the RGC layer of the host retina. Conclusion: This study has established a retinal ganglion cell-specific reporter stem cell line Brn3b-GFP ESC. The GFP-positive RGCs obtained by inducing the cell line into retinal organoids can be integrated into the host retina after transplantation. The establishment of such a cell line will provide an important research tool for glaucoma and related diseases.
视网膜静脉阻塞(retinal vein occlusion，RVO)是发生率仅次于糖尿病性视网膜病变的视网膜血管病。RVO后导致视网膜血管损伤进而引起血管闭塞，造成视网膜缺血从而促进异常视网膜新生血管(retinal neovascularization，RNV)的增生，晚期发生玻璃体积血、新生血管性青光眼等并发症，积极治疗可以稳定患者的眼部情况，避免并发症的发生。本文报告了1例患有高血压病的23岁年轻女性患者发生视网膜分支静脉阻塞并发新生血管增殖膜的病例，给予病变区视网膜激光光凝治疗，10年后随访发现RNV膜机化萎缩。
Retinal vein occlusion (RVO) is the second most common retinal vascular disease after diabetic retinopathy. Retinal vascular damage after RVO leads to venous occlusion, which further causes retinal ischemia and promotes abnormal retinal neovascularization (RNV). Later complications such as vitreous hemorrhage and neovascular glaucoma occur. Active treatment can stabilize the ocular condition of patients and avoid the occurrence of complications. This paper reports a case of a 23-year-old young female patient with hypertension who developed branch RVO complicated by neovascularization membrane. The lesion area was treated with laser photocoagulation. The RNV was found to be mechanized and atrophied after 10 years of follow-up.
目的：分析高度近视有晶状体眼后房型人工晶状体植入术后孔源性视网膜脱离的临床特征及预后。方法：回顾分析2012年4月至2021年6月中山眼科中心收治的9例(9只眼)行后房型人工晶状体植入术后孔源性视网膜脱离患者的临床特征、手术方式及疗效，随访(4.96±4.78)个月。结果：患者年龄(30.44±20.11)岁，屈光手术至发病时间(32.10±17.80)个月。4例(44.4%)马蹄形裂孔，1例(11.1%)萎缩性裂孔，4例(44.4%)巨大裂孔；9眼裂孔均位于赤道部前，除2眼(22.2%)为单个巨大裂孔，1眼(11.1%)单个马蹄孔，余6眼(66.7%)均有视网膜周边变性区存在；视网膜脱离范围(3.0±1.12)个象限，8例累及黄斑；增殖性玻璃体视网膜病变C级以上4眼。视网膜初始复位率为77.8%，最终视网膜复位率100%。末次随访最佳矫正视力优于术前(P<0.05)。随访期间，2例硅油填充眼发生并发性白内障，4眼发生术后早期高眼压。结论：有晶状体眼后房型人工晶状体植入术前存在的视网膜变性或术后玻璃体牵引的存在可能是孔源性视网膜脱离发生的危险因素。
Objective: To analyze the clinical presentation, surgical management, and outcomes of rhegmatogenous retinal detachment (RRD) in patients with high-myopia corrected by posterior chamber phakic (PCP) intraocular lens (IOL) implantation. Methods: Nine eyes of 9 patients in whom RRD developed after PCPIOL implantation from April 2012 to June 2021 in Zhongshan Ophthalmic Center were retrospectively studied. Mean follow-up after retinal detachment surgery was (4.96±4.78)months. Results: Mean patient age was (30.44±20.11) years old. RRD occurred (32.10±17.80) months after PCPIOL implantation. Four (44.4%) breaks were horseshoe tear, 1 (11.1%) was atrophic hole and 4 participants (44.4%) had a giant retinal tear. Nine cases had causative breaks located anterior to the equator while peripheral retina lattice degeneration was found in 6 eyes. RRD extended from 1 to 4 quadrants (3.0±1.12 quadrants) and 8 cases were macula-off retinal detachments. Four eyes’ proliferative vitreoretinopathy were more severe than level C. Initial reattachment rate was 77.80%. Final retinal reattachment was 100%. Final follow-up BCVA was significantly better than baseline (P<0.05). Furthermore, concurrent cataract occurred in 2 eyes in which silicone oil was used as tamponade. Ocular hypertension was detected in 4 eyes after surgery. Conclusion: The existed lattice degeneration and postoperative vitreous traction may be risk factors for RRD after PCPIOL implantation.
目的：以光学相干断层扫描血管成像(optical coherence tomography angiography，OCTA)观察视网膜分支静脉阻塞(branch retinal vein occlusion，BRVO)抗血管内皮生长因子(vascular endothelial growth factor，VEGF)治疗前后的变化。方法：回顾性收集从2017年1月至2018年1 2月在汕头国际眼科中心的确诊为BRVO合并黄斑水肿的患者共3 1例3 2眼。患眼行玻璃体腔注射抗VEGF药物治疗，记录治疗前和治疗后1个月的最佳矫正视力(best corrected visual acuity，BCVA)，OCTA检查视网膜黄斑中心凹厚度(foveal macular thickness，FMT)、黄斑区血流密度。比较治疗前后各指标的变化。结果：治疗后BCVA较治疗前显著提高，差异有统计学意义(P<0.001)；FMT[(242.13±86.02) μm]较治疗前[(521.44±190.27) μm]明显下降，差异有统计学 意义(P<0.001)；中心凹浅层血流密度[(18.44±4.98)%]及中心凹旁浅层血流密度[(44.83±3.19)%]均较治疗前[(25.46±9.21)%，(46.06±5.25)%]相比明显下降，差异有统计学意义(P <0.001)。结论：玻璃体腔注射抗VEGF治疗BRVO合并黄斑水肿效果显著；OCTA能有效评价抗VEGF治疗BRVO合并黄斑水肿的临床疗效。
Objective: To evaluate the efficacy in patients with macular edema due to branch retinal vein occlusion (BRVO) treated with intravitreal anti-VEGF drug. Methods: In this retrospective study, 32 eyes of 31 patients with BRVO combined with macular edema at Joint Shantou international eye center of Shantou University and TheChinese University of Hong Kong during January 2017 to December 2018 were enrolled in this study. All the affected eyes received intravitreal anti-VEGF drug injections. BCVA (best corrective visual acuity) and optical coherence tomography angiography (OCTA) were performed before and one month after intravitreal anti-VEGF drug injections. Foveal macular thickness (FMT), macular blood flow density was measured in all eyes and compared. Results: The BCVA before therapy was (0.77±0.46) LogMAR and increased to (0.46±0.30) LogMAR in one month after therapy, which showed a statistical difference (P<0.001). The FMT, foveal superficial vascular plexus flow density and para foveal superficial vascular plexus flow density before therapy were (521.44±190.27) μm, (21.85±6.17)% and (46.29±2.70)%, respectively. The FMT, foveal superficial vascular plexus flow density and para foveal superficial vascular plexus flow density decreased to (242.13±86.02) μm, (18.40±5.18)% and (44.75±3.40)%, respectively. There was significant statistical difference for them (P<0.001). Conclusion: Intravitreal injection of anti-VEGF is effective in the treatment of BRVO combined with macular edema. OCTA can effectively evaluate the clinical efficacy of anti-VEGF in the treatment of BRVO combined with macular edema.
