更昔洛韦玻璃体腔注药术治疗急性视网膜坏死

目的 探讨更昔洛韦玻璃体腔注药术治疗急性视网膜坏死（ARN）的手术适应证、手术时机及其疗效。方法ARN住院患者14例（14只眼）,均符合美国葡萄膜炎学会ARN诊断标准。患者初诊视力为光感、眼前手动、数指者各1只眼,0．08～0．1者4只眼,0．2～0．4者5只眼,0．5、0．8者各1只眼。角膜后沉着物、房水闪光均阳性。眼底表现为周边部局灶性和（或）片状视网膜坏死、视网膜动脉白线、视网膜出血等。全身分别给予阿昔洛韦或更昔洛韦静脉滴注,患者病情继续发展、恶化,但尚未出现视网膜脱离。再对14只眼行更昔洛韦玻璃体腔注药术。其中2只眼注药后,病情不能控制,出现了增生性玻璃体视网膜病变（PVR）和视网膜脱离,即行玻璃体切除术。术后患者随访4～74个月,平均25个月。结果更昔洛韦玻璃体腔注药术后,12只眼视力显著提高,提高至1．0～1．5者5只眼,0．5～0．9者5只眼,0．3者2只眼。玻璃体切除术后的2只眼,术后视力较术前亦有提高,分别由眼前数指提高至0．4,光感提高至眼前数指。14只眼的眼前节炎性反应和玻璃体混浊消失或明显减轻,视网膜黄白色病变消退,出血吸收,视网膜在位。结论对全身抗病毒药物治疗不能控制病情的ARN患者,在尚未发生PVR或视网膜脱离时,及早给予更昔洛韦玻璃体腔注药术可获得满意疗效,能显著提高患者视力。（中华跟科杂志,2007,43：631-637）     

药物性玻璃体后脱离对增生性玻璃体视网膜病变影响的实验研究

目的 观察药物性玻璃体后脱离（PVD）对增生性玻璃体视网膜病变（PVR）的影响。方法 24只有色成年家兔,左眼为实验眼。兔自体富含血小板血浆玻璃体腔注射建立兔眼PVR模型,同时随机将实验兔分为A、B组及对照组,每组各8只眼。建模后3 h A组玻璃体腔内注入1 U纤溶酶（0.05 ml）+20 U透明质酸酶（0.05 ml）共0.1 ml、B组玻璃体腔内注入纤溶酶0.1 ml、对照组玻璃体腔内注入等量的平衡盐溶液。给药后1、7、28 d记录实验眼PVR级别,进行各组PVR等级评分,并行闪光视网膜电图（F-ERG）、眼部B型超声检查及视网膜组织病理学检查。结果 PVR模型成功建立,并在注药后7 d,A组发生完全性PVD 5只眼,不完全性PVD3只眼;B组不完全性PVD 5只眼,未见完全性PVD,另3只及对照组无PVD发生。A、B组在注药后28 d PVR等级评分均低于对照组,差异有统计学意义（D=75.6, 98.9;P=0.003,P=0.011）;注药后7、28 d,A、B 两组F-ERG b 波波幅均高于对照组;产生PVD的眼中PVR级别均较无PVD的眼级别低,其中完全性PVD眼中PVR级别仅为0～1级。结论 在兔眼PVR建模后3 h,纤溶酶与透明质酸酶联合玻璃体腔注射诱导的完全性PVD在一定程度上可以阻止兔眼PVR的发生和发展,纤溶酶单独或联合透明质酸酶玻璃体腔注射诱导的不完全 性PVD对PVR的发展有减缓作用。     

PDGF抗体和VEGF抗体预防增生性玻璃体视网膜病变

目的评价血小板源性生长因子（PDGF）抗体和血管内皮生长因子（VEGF）抗体对实验性增生性玻璃体视网膜病变（PVR）的预防作用。方法将兔制成PVR模型，将一定浓度的PDGF抗体、VEGF抗体及BSS液分别在当时及第7d注入兔眼的玻璃体腔，每日观察兔眼情况，处死兔子做眼病理切片及免疫组织化学染色。结果间接检眼镜观察可见对照组与实验组均出现视网膜脱离。玻璃体腔内注药后第21d、28dPVR分级实验组与对照组差异有统计学意义（P〈0．05），在制作动物模型时直接注入和第7d时注入差异无统计学意义（P〉0．05）。病理检查所见与间接检眼镜所见相符。免疫组织化学染色计算阳性细胞百分率，亦得出相同结果。结论玻璃体腔内注入PDGF抗体、VEGF抗体均可预防PVR的进展，注药时间对于PVR的发展无明显影响。     

GM6001预防dispase诱导的兔眼增生性玻璃体视网膜病变

目的评价金属蛋白酶抑制剂GM6001对dispase诱导的兔眼实验性增生性玻璃体视网膜病变（PVR）的预防作用。方法健康成年的青紫蓝兔20只，采用dispase诱导的PVR动物模型，右眼玻璃体腔注射100μmol／LGM60010．05ml，左眼注射PBS作为对照，共观察6周。采用PVR分级评分进行临床观察，流式细胞学方法检测玻璃体细胞增生，免疫组织化学方法标记抗增生核抗原（PCNA），判断视网膜细胞增生状况。结果GM6001组与对照组的PVR分级评分分别是1．19和2．54，差异有显著统计学意义（P〈0．05）；GM6001组的玻璃体增生细胞数低于对照组（P〈0．05）；GM6001组PCNA阳性率低于对照组（P〈0．05）。结论GM6001玻璃体腔注射能抑制和延缓实验性PVR的发生。     

增生性玻璃体视网膜病变的药物治疗

增生性玻璃体视网膜病变（PVR）常由于裂孔源性视网膜脱离、眼穿孔伤或眼内手术造成血-视网膜屏障受损,视网膜色素上皮（RPE）细胞进入玻璃体,继而引起RPE细胞、神经胶质细胞、成纤维细胞等在玻璃体内增生,形成以细胞为主的纤维膜。临床上治疗和预防PVR以手术为主,但效果不佳。近来有许多药物治疗PVR的研究报道,就PVR药物治疗研究进展进行综述。     

激肽原-激肽系统参与增生性玻璃体视网膜病变形成的实验研究

背景前期研究发现,激肽原1是增生性玻璃体视网膜病变（PVR）患者玻璃体中的特异蛋白质,是在质谱鉴定中得到的肽段匹配和MASCOT得分最高的蛋白质,且与PVR的严重程度呈正相关。目的探讨激肽原一激肽系统（KKS）是否参与PVR的发生过程。方法大鼠视网膜色素上皮（RPE）细胞系RPE-J细胞复苏培养后用PBS配制成2．5X10^8／ml的细胞悬液,大鼠下腔静脉采血4ml经枸橼酸二钠处理和离心后用PBS制备成血小板密度为2．5×10^8／ml的血浆。用随机数字表法将60只Wistar大鼠随机分为实验组和生理盐水组,每组各30只。实验组大鼠左眼玻璃体腔内注射RPE细胞悬液4ill＋富含血小板血浆6仙l建立PVR模型,生理盐水组左眼玻璃体腔以同样的方法注射生理盐水10μl,各组大鼠的右眼作为自身对照组。术后1、3、7、14、21、28d行裂隙灯及间接检眼镜检查,按Francine的标准进行PVR分级。玻璃体腔注射后28d收集实验动物的玻璃体、血清和视网膜标本,应用Westernblot法检测缓激肽的表达,视网膜标本行组织病理学检查。结果术后28d,实验组有25只眼出现不同程度的PVR表现,建模成功率为89．3％（25／28）。术后7、14、28d裂隙灯下证实实验组大鼠形成1、2、3级PVR。视网膜组织病理学检查表明视网膜组织中炎性细胞浸润及RPE细胞移行并转化为成纤维细胞,出现视网膜脱离。Westernblot分析显示在所有PVR大鼠血清、玻璃体和视网膜中均可检测到缓激肽的表达,但实验组大鼠的表达强度明显高于生理盐水组大鼠。结论玻璃体腔注射RPE细胞联合富含血小板血浆的方法可以成功建立PVR大鼠模型,KKS可能参与PvR的发生。     

羊膜匀浆治疗实验性孔源性视网膜脱离的超微结构观察

目的对羊膜匀浆治疗实验性孔源性视网膜脱离的超微结构进行初步观察,并对其组织病理学结果进行评价。方法20只兔（每只兔1眼）随机分为A组（治疗组）10眼、B组（对照组）10眼。人造视网膜裂孔,治疗组裂孔表面滴加0．1mL羊膜匀浆上清液,对照组滴加0．1mL PBS,术毕20％SF6眼内填充。术后14d处死动物,透射电镜观察。结果术后14d。治疗组视网膜复位6眼（60．00％）,对照组视网膜复位2眼（20．00％）。透射电镜观察,治疗组在视网膜裂孔边缘及底部有多层Maller细胞及RPE细胞增生,与其下脉络膜发生粘连。而对照组,视网膜裂孔边缘细胞增生明显较少。结论羊膜匀浆治疗兔实验性视网膜脱离,有助于封闭视网膜裂孔,裂孔边缘增生细胞经电镜证实主要是视网膜Miiller细胞和RPE细胞     

视紫红质蛋白激酶抑制剂对兔增生性玻璃体视网膜病变的作用

目的 分析视紫红质蛋白激酶（ROCK）抑制剂Y27632对兔增生性玻璃体视网膜病变（PVR）的作用。方法 培养兔视网膜色素上皮（RPE）细胞。相差显微镜及免疫荧光染色检测Y27632对RPE细胞-平滑肌肌动蛋白（SMA）压力纤维形成的影响。体外Ⅰ型胶原凝胶收缩试验和噻唑蓝（MTT）试验来检测Y27632对细胞收缩力量和增殖的影响。用第6代兔RPE细胞建立兔PVR模型,玻璃体腔内给予50μmol／L Y27632,每周1次,持续28d,以牵拉性视网膜脱离的发生率来评估药物的体内效应。行视觉电生理和组织学检查以分析该药的毒性。结果 50μmol／L Y27632破坏了兔RPE细胞α-SMA压力纤维的形成,并减弱了RPE细胞产生的收缩力（P〈0．01）;RPE细胞的增殖不受Y27632的影响。Y27632用药组的牵拉性视网膜脱离的发生率降低（P〈0．01）。在用药组中,未发现明显的视网膜组织结构和功能的损害。结论 特异性ROCK抑制剂Y27632可减弱RPE细胞产生的收缩力,并且可阻止兔PVR的发展。该药无明显的副作用,有可能成为一种防治PVR的有效方法。     

c-fos反义寡核苷酸对兔实验性增生性玻璃体视网膜病变的抑制作用

目的评价c—fos反义寡核苷酸（AS-ON）对兔实验性增生性玻璃体视网膜病变（PVR）的抑制作用。方法将9只兔（18只眼）分为c—fos—AS-ON治疗组和对照组（每组9只眼）。c—fos—AS—ON治疗组以平衡盐溶液（BSS）配制2pgc—fos—AS—ON和10μg脂质体的混合液0．1ml、对照组以0．1mlBSS分别与2．5×10^5个／ml人视网膜色素上皮（RPE）细胞稀释液混合后同时注入玻璃体腔，观察两组在玻璃体腔注射后第7、14、21、28天PVR发生的程度。结果随着观察时间的延长，两组PVR程度逐渐加重。第7天两组PVR程度无统计学差异（P=0．05），第14、21、28天，c—fos—AS-ON治疗组的PVR程度较对照组轻，差异有统计学意义（P〈0．05）；第28天，c—fos—AS—ON治疗组牵拉性视网膜脱离发生率为25．0％，对照组为62．5％。结论c—fos—AS—ON对兔实验性PVR的发生有一定的抑制作用。     

苦参碱聚乳酸微球防治增生性玻璃体视网膜病变的研究

目的评价苦参碱聚乳酸微球防治实验性增生性玻璃体视网膜病变（PVR）的效果。方法30只新西兰白兔玻璃体腔注入成纤维细胞悬液制备PVR模型。随机分为3组,玻璃体腔分别注入载药微球（含苦参碱4mg）、游离苦参碱（2mg）、生理盐水和空白聚乳酸微球。玻璃体腔手术操作后第1、3、7、14、21、28、35天观察眼前节炎症反应情况、玻璃体混浊程度、玻璃体腔内微球分解过程、玻璃体内增生情况及视网膜是否脱离以及脱离的程度。结果所有动物1周内前房有轻～中度的炎症反应,1周后消失。各组均有不同比例的动物在不同时间点发展为PVRⅠ～Ⅲ级。视网膜脱离的发生率：游离药物组与对照组比较,除35d外,其余各时间点差异均有统计学意义（P〈0.05）;载药微球组与对照组相比,各时间点差异均有统计学意义（P〈0.05）;载药微球组与游离药物组比较,28d和35d时差异均有统计学意义（P〈0.05）。结论苦参碱聚乳酸微球兔眼玻璃体腔注射能够有效防治实验性PVR。     

Induction of proliferative vitreoretinopathy by a unique line of human retinal pigment epithelial cells

BACKGROUND: The most widely used models of proliferative vitreoretinopathy (PVR) rely on injection of cells into the vitreous of animals. Using retinal pigment epithelial (RPE) cells from human PVR membranes may produce a more accurate model of human PVR. We performed a study to determine whether human RPE cells derived from a single epiretinal membrane (ERM) are capable of inducing the same disease in the rabbit eye, and whether the induced ERMs had cellular components similar to those of human PVR membranes. METHODS: Cells were harvested from a human ERM obtained at surgery for PVR. RPE cells were cultured from the membrane and injected into the right eye of 24 New Zealand albino rabbits. The left eyes served as controls. The eyes were examined by indirect ophthalmoscopy over 4 weeks. The enucleated eyes were then examined by means of microscopy and histochemical analysis. RESULTS: By day 7, PVR had developed in all but 1 of the 24 experimental eyes, with 8 progressing to localized tractional retinal detachment. By day 21, localized tractional retinal detachment had developed in 17 eyes; 1 eye progressed to extensive tractional retinal detachment by day 28. Immunostaining showed that mostly RPE cells, but also myofibroblasts, glial cells and collagen, were present in the newly formed rabbit PVR membranes. INTERPRETATION: Human RPE cells cultured from a PVR membrane appear to be capable of inducing PVR in rabbits. The resultant ERMs are similar to those formed in human PVR and consist mainly of RPE cells.     

Evaluation of experimental proliferative vitreoretinopathy in rabbits

We established an effective animal experimental model for proliferative vitreoretinopathy (PVR) by intravitreally injecting cultured cells in order to investigate therapies for PVR, including intravitreal drugs, radiation and hyperthermia. After making posterior vitreal separation by injecting SF6 gas into the rabbit's vitreous body, cultured cells were intravitreally injected. Fundus changes were followed up for 4 weeks and PVR stage was recorded according to the classification described by Hida et al. The cultured cells injected were rabbit dermal fibroblasts (5 x 10(4) and 10 x 10(4) and human retinal pigment epithelial (RPE) cells (5 x 10(4]. A total of 37% of animals reached STAGE 5-7 (traction retinal detachment) in 4 weeks in 41 eyes injected with 5 x 10(4) rabbit fibroblasts and were 86% in 14 eyes injected with 10 x 10(4) rabbit fibroblasts. The 10 eyes injected with human RPE cells showed STAGE 2 or less in 4 weeks. Consequently, we selected the experimental PVR model using injecting of 10 x 10(4) rabbit fibroblasts as the most effective method.     

Quantitative assessment of growth stimulating activity of the vitreous during PVR.

This report postulates that the activity of cellular proliferation along the surface of the retina during proliferative vitreoretinopathy (PVR) is reflected in the aggregate effect of proliferation-inducing growth factors in the vitreous. A method for quantifying the "net" proliferation-inducing capacity of an individual vitreous sample was developed and this assay was used to evaluate the vitreous proliferative activity in a model for experimental PVR. Vitreous was aspirated sequentially after onset of fibroblast-induced PVR in a rabbit model. A simple bioassay for the "aggregate" stimulating activity was developed and each sample was assigned a quantitative value in terms of "proliferation units" (PU). Experimental eyes demonstrated a wide range of stimulating activity (0-1765 PU), but control eyes showed uniformly low levels of activity (0-337 PU). Experimental eyes that ultimately developed retinal detachment displayed higher levels of proliferative activity than did those eyes destined to remain attached. The differences were statistically different by day 3, prior to the onset of clinical retinal changes. We conclude that quantification of vitreous proliferation-stimulating activity is possible and that this method might be useful for screening eyes at high risk for the development of recurrent retinal detachment from PVR.     

The development of severe proliferative vitreoretinopathy after retinal detachment surgery

A prospective clinical study was conducted to determine wether preoperative proliferative vitreoretinopathy (PVR), grade B, was a significant risk factor in the development of severe PVR after surgery for retinal detachment repair. Two series of consecutive retinal detachments associated with horseshoe retinal tears were compared. The first series included 40 eyes of 40 patients with preoperative PVR, grade O - A. The second series included 30 eyes of 27 patients with preoperative PVR, grade B. All eyes were operated on with conventional microsurgical techniques. At the first operation, no vitrectomies were carried out in any eyes. The incidence of postoperative PVR, grades C and D, was 20% (6/30 eyes) after a single operation in the series of eyes with preoperative PVR, grade B as compared to 0% in the series of eyes with preoperative PVR, grade O - A. The difference between the two groups was statistically significant (P = 0.01). It was also found that the incidence of postoperative proliferative PVR was significantly higher in eyes with preoperative vitreous hemorrhage (30.7%) as compared to eyes with no preoperative vitreous hemorrhage (0%;P = 0.02). Incomplete posterior vitreous detachment without collapse of the vitreous gel occurred significantly more frequently in eyes with preoperative proliferative vitreoretinopathy, grade B (68.4%, than in eyes with preoperative proliferative vitreoretinopathy, grade O - A(27.5%;P = 0.02).     

视网膜脱离合并脉膜脱离的手术治疗

目的：探讨视网膜脱离合并脉膜脱离的手术方法、效果及失败原因,全部眼有脉络膜脱离和增殖性玻璃体视网膜病变（VR）,采用玻璃体切割及视网膜前膜,眼内惰性气或硅油填充。结果：31例患视网膜全部复位者22例（70．9％）部分复位5例（16．1％）,未复位4例（12．9％）。25眼和有增进,5眼无变化,1眼下降。手术失败的主要原因可能与严重的前部PVR有关,结论：伴有脉膜脱离的视网膜脱离患者采用VR手术,可提高其手术成功率。     

增生性玻璃体视网膜病变中组织型转谷氨酰胺酶的表达

目的:检测增生性玻璃体视网膜病变(PVR)视网膜前膜和视网膜色素上皮(RPE)细胞中组织型转谷氨酰胺酶(tTG)的表达情况,探讨tTG是否参与PVR的发病过程.方法:PVR患者21例,收集视网膜前膜,C级8例,D级13例,行tTG免疫组织化学染色.另对培养3-5代的人RPE细胞分别用含有1 00mL/L PVR玻璃体液、正常玻璃体液和PBS的DMEM培养液进行孵育24h后行tTG免疫细胞化学染色.光镜下观察,比较C级和D级膜的表达阳性率,显微图像分析测量3组RPE细胞染色的平均灰度值.结果:在PVR视网膜前膜中tTG呈阳性表达(81%),C级和D级膜表达阳性率无差异 (C级88%,D级77%,P＞0.05).培养的人RPE细胞表达tTG,在PVR患者的玻璃体液作用下,tTG表达的平均灰度值为137.0±2.6,与正常玻璃体液组(143.5±2.9)及PBS对照组(143.6±3.0)相比,表达水平升高(P＜0.05).结论:PVR视网膜前膜和培养的人RPE细胞tTG表达阳性,在PVR患者的玻璃体液作用下,RPE细胞的tTG表达水平升高,这表明tTG参与了PVR的发病过程,在PVR视网膜前膜的形成过程中可能有重要作用.     

结缔组织生长因子在兔视网膜增生膜中的表达

目的观察结缔组织生长因子（CTGF）在实验性增生性玻璃体视网膜病变（PVR）增生膜中的表达,探讨CTGF在PVR视网膜增生膜形成过程中的作用。方法采用Nobuyo的方法从兔视网膜中分离视网膜色素上皮（RPE）细胞并进行体外培养和传代。第3代RPE细胞制备密度为1.1×10^5/mL的细胞悬液。32只日本大耳白兔,取8只作为正常对照组,其余24只采用玻璃体腔内注入RPE细胞悬液的方法建立PVR动物模型。动物分别于造模后7、30、60d处死并摘除眼球,每次处死8只动物。光学显微镜下观察视网膜增生膜的组织学改变,免疫组织化学法检测PVR视网膜增生膜中CTGF的表达。结果造模后5d检眼镜下可见视网膜增生组织开始形成,增生膜随时间的推移逐渐增厚。组织学检查显示,造模后7d兔视网膜表面可见红染的条索状和网状胶原纤维,并可见大量的增生细胞分布其中。光学显微镜下可见视网膜内界膜变厚、粗糙、断裂或结构不清,视网膜各层结构欠清。免疫组织化学法检测表明,正常对照组在玻璃体及视网膜内未见CTGF的特异性染色。造模后7d,CTGF主要表达于视网膜表面增生细胞;造模后30d,CTGF主要表达于增生的细胞及增生的胶原纤维组织中;造模后60d,CTGF主要表达于增生的胶原纤维组织中。结论 CTGF在实验性PVR动物模型中呈高表达,提示CTGF参与了PVR增生膜的形成。     

Gravity-dependent distribution of retinal pigment epithelial cells dispersed into the vitreous cavity

Release of viable retinal pigment epithelial (RPE) cells into vitreous cavity and subsequent attachment to the retina may be the first steps in the occurrence of macular pucker and proliferative vitreoretinopathy (PVR) complicating rhegmatogenous retinal detachment. Gravity and post-operative position of the patient together may influence where the cells settle, and, thereby, the location of subsequent membrane formation. To study the effect of gravity on the location of RPE cell attachment, 3H-thymidine-labelled RPE cells were injected into 12 enucleated pig eyes after vitrectomy performed to create a posterior vitreous detachment. The eyes were then positioned to make either the macula or the inferior retina gravitationally dependent. Radioactivity was later measured from several locations on the retina to indicate the proportion of cells attached in each location. Radioactivity measured from the dependent part of the globe (mean 5985 +/- 1728) was always greater than that from other parts (mean 389 +/- 79). The experiment was repeated in live pig eyes with identical results. These findings suggest that patient positioning may affect the location of cellular membrane formation and subsequent retinal traction.