目的：测量黄斑区视网膜神经节细胞层和内界膜-视网膜色素上皮层厚度，分析其与新发缺血性脑卒中的相关性。方法：采用前瞻性研究，选择2021年12月至2022年3月在潍坊医学院附属医院体检的健康者30例(60眼)为对照组(A组)；选择同期收治且年龄匹配的新发缺血性脑卒中患者30例(60眼)为卒中组(B组)。比较两组黄斑区神经节细胞层(ganglion cell layer,GCL)和内界膜-视网膜色素上皮层(inner limiting membrane-Retinal pigment epithelium,ILM-RPE)厚度及平均GCL厚度/ILM-RPE平均容积厚度(GCL/ILM-RPE厚度比)。结果：共纳入120只眼，A、B组分别60只眼。A组的平均GCL为(83.5±4.7)μm，B组的平均GCL厚度为(78.7±10.5)μm，组间比较差异有统计学意义(P<0.001)。A组的GCL最小厚度为(80.4±4.4)μm，B组的GCL最小厚度为(70.7±16.3)μm，组间比较差异有统计学意义(P<0.001)。A组的ILM-RPE中心子区厚度为(247.5±19.8)μm，B组的ILM-RPE中心子区厚度为(241.6±36.3)μm，组间比较差异无统计学意义(P=0.393)。A组的ILM-RPE平均容积厚度为278.4±9.8μm，B组的ILM-RPE平均容积厚度为(278.5±15.3)μm，组间比较差异无统计学意义(P=0.448)。A组的GCL/ILM-RPE厚度比为0.300 3±0.012 2，B组的GCL/ILM-RPE厚度比为0.286 2±0.028 6，组间比较差异有统计学意义(P<0.001)。结论：缺血性脑卒中影响GCL厚度和GCL/ILM-RPE厚度比， GCL厚度和GCL/ILM-RPE厚度比与新发缺血性脑卒中可能有关。
Objective: To measure the thickness of retinal ganglion cell layer and inner limiting membrane-retinal pigment epithelium layer in the macular region, and analyze its correlation with new-onset ischemic stroke. Methods: By using a prospective study, 30 healthy subjects (60 eyes) who underwent physical examination in Affiliated Hospital of Weifang Medical University from December 2021 to March 2022 were selected as the control group (Group A). Thirty age-matched patients with new-onset ischemic stroke admitted at the same time (60 eyes) were selected as the stroke group (Group B). The ganglioncelllayer (GCL) and innerlimitingmembrane-Retinalpigmentepithelium (ILM-RPE) were compared between the two groups. It included the thickness and mean GCL thickness/Mean Volume thickness of ILM-RPE (GCL/ILM-RPE thickness ratio). Results: A total of 120 eyes were included, with 60 eyes in group A and B. The average GCL thickness in group A was (83.5±4.7) μm, and the average GCL thickness in group B was (78.7±10.5) μm,and the difference between groups was statistically significant (P<0.001). The minimum thickness of GCL in group A was (80.4±4.4) μm, and the minimum thickness of GCL in group B was (70.7±16.3) μm, showing statistical significance(P<0.001). The thickness of the central subregion of ILM-RPE in group A was (247.5±19.8) μm, and the thickness of the central subregion of ILM-RPE in group B was (241.6±36.3) μm, and the difference was not statistically significant between two groups (P=0.393). The mean volumetric thickness of ILM-RPE in group A was (278.4±9.8) μm, and the mean volumetric thickness of ILM-RPE in group B was (278.5±15.3) μm, and the difference was not statistically significant (P=0.448). The GCL/ILM-RPE thickness ratio of group A was (0.300 3±0.012 2), and that of group B was (0.286 2±0.028 6), showing a statistical difference between groups (P<0.001). Conclusion: Ischemic stroke affects the GCL thickness and the thickness ratio of GCL/ILM-RPE, which may be related to new-onset ischemic stroke.
目的：借助于人工智能(artificial intelligence,AI)眼底筛查远程接转诊系统，探索“患者-社区-医院”远程筛查模式，推进眼科分级诊疗和双向转诊实施，为地市级医疗机构开展眼底疾病人工智能筛查工作提供一定的经验借鉴。方法：通过AI辅助远程筛查基层医疗机构的4886例患者，完成眼科检查并经AI初判、人工复核形成眼底诊断结论。通过医联体和专科联盟模式，对基层医疗机构的4886例患者的AI诊断系统结果和上级医师审核结果进行对照分析，分析AI诊断系统在眼科常见病种筛查中的推广应用的可信度和可行性。结果：AI检出DR的灵敏度为94.70%，特异度96.06%；DME的灵敏度96.43%，特异度96.55%；AMD的灵敏度77.55%，特异度95.74%；同时，其在病理性近视、白内障、青光眼等常见病种眼底筛查中也有一定作用。结论：AI辅助远程筛查系统对于绝大多数眼底疾病有较高的敏感性和特异性，适用于眼底疾病的筛查工作，利于基层医院或社区医院对于眼底疾病的初步诊断，落实眼科分级诊疗，有借鉴推广意义。
Objective: With the help of artificial intelligence (AI) based fundus screening remote referral telemedicine system,it enables us to explore the remote screening mode of patient-community-hospital, and promote the two-way referral and ophthalmic graded diagnosis. This investigation provides certain practice experiences for prefecture-level medical institutions to carry out AI screening for fundus diseases. Methods: Ophthalmologic examination was performed on 4,886 patients in primary medical institutions through AI-aided remote screening, and the final fundus diagnosis conclusion was formed after AI preliminary judgment and manual review. Through the Medical Consortium and specialty alliance model, the results of the AI diagnosis system and the audit results of superior physicians for 4 886 patients in primary care institutions were compared and analyzed, and the credibility and feasibility of the AI diagnosis system application in the screening of common ophthalmic diseases were discussed. Results: The sensitivity and specificity of AI detection of diabetic retinopathy were 94.70% and 96.06%, respectively. In the diabetic macular edema classification, the sensitivity and specificity were 96.43% and 96.55%, respectively. In the age-related macular degeneration classification, the sensitivity and specificity were 77.55% and 95.74%, respectively. Meanwhile, it also plays a role in screening common fundus diseases such as pathological myopia, cataract and glaucoma. Conclusion: The AI-aided remote screening system has high sensitivity and specificity for most of fundus diseases, indicating it is promising for fundus diseases screening in primary medical institutions. It is conducive for primary hospitals or community hospitals to carry out the initial diagnosis of fundus diseases, as well as the implementation of graded diagnosis and treatment of ophthalmology, which has reference and promotion significance.
目的：应用Cirrus HD-OCT 5000对正常眼黄斑及视盘血流参数进行血管成像测量，评估其可重复性及再现性。方法：纳入40只正常眼进行前瞻性研究。操作者A于一周内3天的同一时段(T1，T2，T3)对正常受试者的同一眼黄斑及视盘进行3次血管成像扫描，扫描过程中均开启FastTracTM图像跟踪功能，操作者B在T 2时间点再次对受试者同一眼进行相同程序扫描，使用Angio PlexMetrixTM量化软件(版本10.0)自动测量黄斑和视盘的血管长度密度(vessel length density，VD)和血管灌注密度(vascular perfusion density，PD)。应用单因素方差分析或非参数检验比较3次扫描的VD、PD是否有差异。采用组内相关系数(intraclass correlation coefficient，ICC)、变异系数(coefficient of variation，CV)评价操作者A的可重复性，采用一致性相关系数(consistent correlation coefficient，CCC)、重复性系数(repeatability coefficient，CR)、C V评价操作者间的再现性。结果：操作者A对黄斑及视盘各区域3次扫描的V D、P D之间差异无统计学意义(P >0.05)。操作者A对黄斑和视盘V D、PD 3次扫描的ICC值分别为0.260~0.517、0.362~0.898，黄斑及视盘各区域V D、P D的C V值均<8.1%。黄斑和视盘V D、P D的操作者间CCC值分别为0.3130~0.5665、0.5149~0.7801；黄斑VD和PD的CR值分别为3.2212~4.6399、0.0574~0.0832；视盘VD和PD的CR值分别为2.0675~4.0630、0.0447~0.0730。黄斑CV值均<9.0%，视盘CV值均<6.9%。结论：非同日的同一时段视盘浅层血流参数具有较好的重复性及再现性，黄斑浅层血流参数的重复性及再现性相对较差。
Objective: To evaluate the repeatability and reproducibility of quantification of macular and peri-papillary superficial capillaries in normal eyes using Cirrus HD-OCT 5000 angiography. Methods: Forty normal eyes were included in this prospective study. Angiography 3 mm × 3 mm scan protocol centered on the disc or macula of the same eye was repeated 3 times by operator A in the same period on 3 days (T1, T2, T3) within 1 week,and the FastTracTM image tracking function was turned on during the scan. Operator B repeated the same scan independently at T2 after operator A. Angio Plex MetrixTM quantification software (version 10.0) was used to automatically measure the vessel length density (VD) and vascular perfusion density (PD) of macular and peri-papillary superficial capillaries. One-way analysis of variance or nonparametric test was used to compare the VD and PD measurements of the 3 scans. The intraclass correlation coefficient (ICC) and coefficient of variation (CV)were calculated to evaluate the intra-operator repeatability. Consistent correlation coefficient (CCC), repeatability coefficient (CR), and CV were calculated to evaluate the inter-operator reproducibility. Results: There was no statistically significant difference in the measurements of VD and PD between the 3 scans of the macular and peri-papillary superficial capillaries by operator A (P>0.05). The ICC values for the 3 scans of VD and PD of the macular and peri-papillary superficial capillaries by operator A were 0.260–0.517 and 0.362~0.898, respectively.The CV values of the VD and PD of the macular and peri-papillary superficial capillaries were all <8.1%. The inter-operator CCC values of the macular and peri-papillary superficial capillaries were 0.3130–0.5665 and 0.5149~0.7801,respectively; the CR values of the VD and PD of macula were 3.2212–4.6399 and 0.0574–0.0832, respectively; the CR values of the VD and PD of optic disc were 2.0675–4.0630 and 0.0447–0.0730, respectively. The CV values of macular were all <9.0%, and the CV values of optic disc were all <6.9%. Conclusion:The macular and peri-papillary superficial capillaries parameters in the same period on different days have a good repeatability and reproducibility,while the superficial macular blood flow parameters have relatively poor repeatability and reproducibility.
目的：探讨原发性慢性闭角型青光眼(chronic primary angle-closure glaucoma，CPACG)患者的视网膜血流密度(vessel density，VD)与视野缺损程度的相关性。方法：光学相干断层血管成像技术(optical coherence tomography angiography，OCTA)测量89例(112眼)视野缺损的CPACG患者的黄斑区VD、视盘旁VD，分析VD与视野缺损程度的相关性。结果：视盘旁VD与视野缺损程度成负相关(r>–0.728，P<0.05)。黄斑浅层总VD的受试者工作曲线(receiver operating characteristic，ROC)及曲线下面积(area under the curve，AUC)为0.874。在控制年龄、眼压及视力的情况下，黄斑总浅层VD每降低1%，视野平均缺损(mean deviation，MD)值增加–0.639 dB。结论：CPACG患者VD与视野缺损呈线性负相关，OCTA可以方便无创地观察青光眼患者眼底血流情况，在视野缺损前发现视网膜VD降低，从而可以作为CPACG早期诊断的参考指标。
Objective: To investigate the correlation between the retinal vessel density (VD) and the degree of visual field loss in chronic primary angle-closure glaucoma (CPACG). Methods: Eighty-nine CPACG patients (112 eyes)with different degrees of visual field loss were measured with optical coherence tomography angiography (OCTA) for macular VD and para-optic microcirculation VD, and the correlation between them and the degree of visual field defect were analyzed. Results: There was a negative correlation between the VD of the microcirculation in each zone next to the optic disc and the degree of visual field loss (r>–0.728, P<0.05). The receiver operating characteristic (ROC) and area under the curve (AUC) of the total VD of the superficial macula is 0.874. Under the condition of controlling age, intraocular pressure and vision, for every 1% decrease in the total superficial macular VD, the average visual field defect mean deviation (MD) value increases –0.639 dB. Conclusion: The VD of CPACG patients is linearly negatively correlated with visual field defects. OCTA can conveniently and non-invasively observe the blood flow of the fundus in patients with glaucoma. It is found that the retinal VD is reduced before visual field defects, which can be used as a reference index for early diagnosis of CPACG.
抗血管内皮生长因子(vascular endothelial growth factor，VEGF)治疗视网膜静脉阻塞(retinal vein occlusion，RVO)继发黄斑水肿(macular edema，ME)的有效性及安全性已得到广泛证实。但抗VEGF治疗方案尚无统一标准。现行的治疗方案主要包括固定治疗方案、按需(pro re nata，PRN)治疗方案、稳定性标准驱使的按需(stabilization criteria-driven PRN)治疗方案、治疗与延长(treat and extend，T&E)方案。近年来不少研究综合比较了各个治疗方案在改善视功能、量化评估疾病活动性、调整随访频率等多个维度的表现，为临床医生提供选择抗VEGF治疗方案的参考依据。本文旨在回顾并总结近年来对抗VEGF药物治疗RVO继发ME的研究，阐述抗VEGF治疗方案的研究进展。
The efficacy and safety of anti-vascular endothelial growth factor (VEGF) in the treatment of macular edema (ME) secondary to retinal vein occlusion (RVO) have been widely confirmed. However, there is no unified standard for anti-VEGF treatment regimens. Current treatment regimens mainly include fixed treatment regimen, pro re nata (PRN) treatment regimen, stabilization criteria-driven PRN treatment regimen, and treat and extend (T&E) regimen. In recent years, many studies have compared different treatment regimens in composite dimensions,including improving visual function, assessing disease activity quantitatively and adjusting the follow-up frequency,to provide clinicians with a reference of choosing anti-VEGF treatment regimens. The purpose of this article is to review and summarize recent researches on anti-VEGF drugs in the treatment of ME secondary to RVO, and to clarify the research progress in the anti-VEGF treatment regimens.
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.
