PTEN mRNA在兔晶状体上皮细胞增殖中的表达

目的：探讨PTEN mRNA在兔晶状体上皮细胞增殖中的作用。方法：将健康白色家兔42只随机分为实验组（36只）和对照组（6只）。实验组兔行双眼透明晶状体皮质吸除术,对照组未做处理。在术后ld,3d,1wk,2wk,1mo和2mo各处死6只。应用免疫组化法检测赤道部晶状体上皮细胞中增殖细胞核抗原（proliferating cell nuclear antigen,PCNA）;应用原位杂交和RT-PCR方法检测PTEN mRNA的表达。结果：①正常赤道部晶状体上皮细胞核可见PCNA弱阳性表达,术后1～7d PCNA的表达维持在高水平状态。2wk后,随着时间的延长,PCNA表达逐渐减弱,1～2mo时大致恢复术前状态。②正常兔晶状体上皮细胞中PTEN mRNA的表达呈阳性,术后1d时PTEN mRNA表达明显下降;术后3d时,PTEN mRNA表达仍处于低水平状态。随着时间的延长,术后1wk时略有恢复,2wk时趋于明显好转,1-2mo时恢复正常。PTEN mRNA与PCNA的表达呈负相关。结论：PTEN mRNA在正常兔晶状体上皮细胞胞质中有表达;PTEN参与晶状体上皮细胞的增殖代谢过程,对细胞的增殖起抑制作用。     

细胞浆信息传递介质Smad 4在兔晶状体上皮细胞增殖中的作用

目的：探讨细胞浆信息传递介质Smad4在兔晶状体上皮细胞增殖及转化中的作用。方法：将健康白色家兔63只随机分成实验组（49只）及对照组（14只）,实验组兔双眼行透明晶状体皮质吸除术,对照组兔不做处理。在术后1及3d,1周、1、2、3及5个月分别处死7只实验组兔及2只对照组兔,分别应用原位杂交及免疫组化技术检测晶状体上皮细胞中Smad 4mRNA及增殖细胞核抗原（proliferating cell nuclear antigen,PCNA）的表达。结果：正常兔晶状体上皮细胞胞浆中Smad 4mRNA表达呈弱阳性。术后1d晶状体赤道部及前囊下部Smad 4mRNA无表达,吸光度（A）值与术前相比,差异有显著意义（P＜0．01）;术后3d Smad 4mRNA呈阳性表达,且随时间延长表达逐渐增强,晶状体赤道部细胞在术后1周时Smad 4mRNA表达最强;晶状体前囊下部细胞在术中1个月时Smad 4mRNA表达最强。PCNA的表达在术后1d最强,之后逐渐减弱,术后1个月时恢复至术前水平。术后1－2个月,晶状体赤道部出现转化的成纤维细胞,Smad 4mRNA呈阳性表达。结论：Smad 4mRNA在正常晶状体上皮细胞胞浆中有表达,Smad 4参与晶状体上皮细胞的增殖代谢过程,可能促使晶状体上皮细胞转化为成纤维细胞。     

兔晶状体皮质吸出后FGF-R1 mRNA在细胞增殖中的作用

目的：探讨碱性成纤维细胞生长因子受体1(fibroblast growth factor receptor 1，FGF-R1)在兔晶状体上皮细胞增殖中的作用。方法：将健康白色家兔42只随机分入实验组(36只)和对照组(6只)。实验组兔行双眼透明晶状体皮质吸除术，对照组未做处理。在术后1，3，7，14，30，60d各处死6只。应用免疫组化和RT-PCR方法检测赤道部晶状体上皮细胞中增殖细胞核抗原(proliferating cell nuclear antigen，PCNA)及FGF-R1 mRNA的表达。结果：FGF-R1 mRNA相对含量随术后时间的发展，表现出明显的变化。术后1～3d时FGF-R1表达迅速升高到最高值。术后7d时仍维持在较高水平状态。14d后明显下降，逐渐恢复术前状态。FGF-R1 mRNA与：PCNA的表达呈正相关。结论：FGF-R1参与兔晶状体上皮细胞的增殖代谢过程，与细胞的增殖状态相关。     

环孢素A对兔晶状体上皮细胞增殖过程中磷酸肌醇-3激酶途径的影响

目的:研究环孢素A对白内障术后晶状体上皮细胞增殖过程中磷酸肌醇-3激酶(phosphatidylinositol-3 kinase,PI-3 k)途径的影响,为后发性白内障的治疗提供实验依据。方法:将健康白色家兔30只60眼,行双眼透明晶状体皮质吸除术,右眼为治疗组,左眼为对照组。术后第1d起,对照组眼用生理盐水点眼6次,而治疗组眼应用1%环孢素A(cyclosporine A,Cs A)眼药水点眼6次。分别在术后第1d未点药前、术后1wk,2wk,1mo和2mo各处死随机选择的6只兔,摘取双眼球。应用免疫组织化学、原位杂交方法检测赤道部晶状体上皮细胞中增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)及张力蛋白同源的28号染色体缺失磷酸酯酶mRNA(PTEN mRNA)和Ser473-R的表达。结果:术后两组PCNA和Ser473-R的表达均逐渐降低,其中1wk(0.690±0.035 vs 0.785±0.015,t=6.099,P<0.01)和2wk(0.571±0.038 vs 0.670±0.037,t=4.585,P<0.01)时治疗组PCNA表达均显著低于对照组。另外,1wk(0.374±0.031 vs 0.435±0.030,t=3.486,P=0.006)和2wk(0.220±0.022 vs 0.251±0.020,t=2.516,P=0.031)时治疗组Ser473-R的表达均显著低于对照组。术后1wk~1mo时,两组PTEN mRNA均逐渐回升。术后1wk,治疗组A值显著高于对照组(0.302±0.027 vs 0.255±0.038,t=2.474,P=0.033)。结论:环孢素A对兔眼白内障术后晶状体上皮细胞增殖过程中PI-3k途径有显著抑制作用,为研究CsA抑制后发性白内障提供实验依据。     

去整合素kistrin干预兔后发性白内障形成中晶状体上皮细胞FAK及ERK基因表达的研究

目的研究去整合素kistrin对兔后发性白内障模型晶状体上皮细胞中黏着斑激酶(focal adhesion kinases,FAK)mRNA和其下游因子细胞外信号调节激酶(extracellular signal regulated kinase,ERK)mRNA表达的影响,以探讨kistrin降低兔后发性白内障形成的可能分子机制。方法 8只新西兰大白兔均行右眼透明晶状体囊外摘出术(extracapsular lens extraction,ECLE)建立后发性白内障模型。随机将8只术眼分为实验组和对照组(每组4眼),术毕实验组囊袋内注入80μg.L-1kistrin0.2mL,对照组注入等量林格氏液。随机选4眼左眼为空白组。术后14d取残留的晶状体囊袋行RT-PCR检测FAK和ERK mRNA的表达。结果正常兔晶状体上皮细胞中(空白组)FAK和ERKmRNA表达量少,分别为1.476±0.802、0.576±0.035。ECLE术后14d对照组FAK和ERK mRNA表达量分别为6.496±1.501、1.158±0.573,实验组分别为6.060±1.512、1.262±0.329,2组表达量均较空白组显著性升高,差异均有统计学意义(均为P<0.05),实验组与对照组比较差异均无统计学意义(均为P>0.05)。结论兔后发性白内障形成的早期,FAK及ERK基因表达上调。应用kistrin未改变此两基因在转录水平上的表达。     

糖尿病兔后发性白内障发生过程中晶状体上皮细胞增殖的变化

目的观察糖尿病兔后囊膜混浊(posterior capsular opacification,PCO)发生过程中晶状体上皮细胞(lens epithelial cells,LEC)的增殖情况,以探讨高血糖下LEC增殖的变化规律。方法 24只12～16周龄雄性新西兰大白兔建立1型糖尿病模型后行右眼晶状体囊外摘出术,术后选取1d、7d、10d、12d及2周、4周、6周、8周8个时间点在裂隙灯显微镜下观察PCO程度,并用Odrich法对PCO进行分级,每个时间点取3只兔眼,检测增殖细胞核抗原在后囊膜LEC中的表达,并换算成增殖指数(proliferative index,PI)值。结果糖尿病兔术后10d时可观察到部分标本出现后囊膜轻度混浊;6周时可观察到完全PCO,差异有统计学意义(P<0.05)。术后1d、7d PI值均为0,10d、12d及2周、4周、6周、8周时的PI值分别为0.320±0.020、0.520±0.020、0.753±0.306、0.907±0.012、1.000±0.000、0.947±0.012,术后10d到6周每个时间点PI值与上一时间点比较差异均有统计学意义(均为P<0.05),术后6周PI值达到高峰,8周时开始下降。结论糖尿病兔晶状体摘出术后随着时间的推移PCO形成逐渐加重,术后10d LEC开始发生增殖,6周时达到高峰。     

阿霉素抑制兔眼晶状体上皮细胞增殖细胞核抗原表达的实验研究

目的 探讨人工晶状体植入术后阿霉素(adriamycin，ADM)抑制兔眼晶状体上皮细胞(rabbit lens epithelial cells，RLEC)增殖的作用及客观评价后囊膜混浊(posteriorcapsule opacification，PCO)的方法。方法 采用兔眼品状体囊外摘出联合人工晶状体植入术的动物模型，术中将携带ADM药物缓释系统植入l5只兔眼晶状体囊袋内。术后观察后囊膜混浊情况；术后第4周，链霉亲和素免疫组化法(strePt—avidin—biotin—enzyme comple，SABC)检测兔眼品状体上皮细胞的增殖细胞核抗原(pro1iferating cell nuclear antigen，PC—NA)的表达。结果 实验组与对照组比较，兔眼晶状体上皮细胞的PCNAP阳性表达下降，2组比较差别具有显著性意义。结论 ADM能有效抑制人工晶状体植入术后RLEC的增殖，免疫组化技术(SABC法)检测RLEC的PCNA的表达，为实验室客观评价PCO提供一种简便有效的方法。     

兔眼IOL术后基质金属蛋白酶抑制因子在虹膜、晶状体上皮细胞的表达

目的　探讨白内障囊外摘出及人工晶状体植入术后基质金属蛋白酶抑制因子 (TIMPs)在虹膜、晶状体上皮细胞的表达和TIMPs对后囊膜混浊的形成及纤维化的影响。方法　取 2 5只健康成年家兔 ,均一只眼行晶状体囊外摘出及人工晶状体植入术 ,另一眼作为对照组。每 5只兔眼为一组 ,分别于术后 1、3、7、14、3 0d取出虹膜和晶状体上皮细胞 ,用RT PCR和反向酶谱分析法检测各标本中的TIMPsmRNA和蛋白质的表达 ,并用羟脯氨酸试剂盒检测晶状体囊膜羟脯氨酸量的变化。结果　在正常虹膜、晶状体上皮细胞组织均有TIMP 1、 2、 3和 4mRNA的表达 ,而无相对蛋白质活性的表达 ;术后第 1d ,TIMP 1、 2、 3和 4mRNA即出现明显升高 ,其中术后第 7d ,TIMP 1和 2mRNA的表达量为最大 ,此后逐渐下降 ,术后第 3 0d的表达量仍高于对照组 (P <0 0 5 ) ;TIMP 3mRNA轻度升高 ,TIMP 4mRNA则轻度下降 ;羟脯氨酸的含量于术后 1、3、7d和 14d明显低于术后 3 0d(P <0 0 5 )。结论　TIMPs可能是抑制白内障囊外摘出及人工晶状体植入术后细胞外基质降解的主要因素 ,还可能是后囊膜混浊的形成和纤维化的重要原因之一。     

阿霉素防治兔后囊混浊的实验与临床研究

目的研究阿霉素(adriamycine,ADR)对兔后发性白内障形成的影响及对兔眼的毒性作用.方法在12只兔眼晶状体囊外摘出术中分别注入0.2mLADR(0.4mg@L-1),然后观察术后不同时间角膜及晶状体后囊混浊变化,并结合观察术后病理学和电镜结构变化.结果12只兔眼注入ADR后1周炎性反应比对照组稍重,术后3～8周后囊膜明显比对照组清晰.组织病理学检查表明,对照组术后晶状体上皮细胞增生,而用药组明显抑制晶状体上皮细胞增生,且对角膜无明显毒性.ADR引起晶状体上皮细胞空泡变性、核固缩等,电镜观察发现用药组晶状体上皮细胞体积缩小、胞浆浓缩、染色质靠边聚集、包膜完整等典型的细胞凋亡形态.结论ADR能有效地抑制晶状体上皮细胞的增殖,预防兔后发性白内障的形成,且对兔眼毒性小,是-种有效的防治后发性白内障的药物.     

兔眼超声乳化吸出术后非手术眼房水及血清中TNF-α、IL-1β表达变化与该眼角膜知觉敏感度变化之间的关系

目的　探讨超声乳化吸出术后不同时间点兔非手术眼房水及血清中肿瘤坏死因子α（TNF-α）和白细胞介素-1β（IL-1β）表达变化与该眼角膜知觉敏感度变化之间的关系。方法　选取健康成年新西兰大白兔40只作为实验对象。采用随机数字表法将大白兔分为实验组与空白对照组,实验组25只,空白对照组15只。实验组:一眼行晶状体超声乳化吸出术,根据术后取材时间分为术后1 d、3 d、7 d、14 d、21 d共5组,每组5只动物。空白对照组:按照对应取材时间也分为5组,每组3只动物。术后观察并记录各组兔双眼的结膜充血、角膜混浊及前房炎症反应情况。实时定量PCR检测各组兔血清TNF-α mRNA和IL-1β mRNA表达水平,ELISA法检测房水中TNF-α、IL-1β蛋白浓度,角膜知觉计测量角膜知觉敏感度。结果　实验组非手术眼与空白对照组双眼在不同时间点,结膜充血、角膜混浊及前房炎症反应级别均为0级。术后1 d、3 d、7 d、14 d,实验组兔血清TNF-α mRNA和IL-1β mRNA表达水平均较基线值升高,术后7 d达高峰（TNF-α mRNA相对表达量为14.95±0.89,IL-1β mRNA相对表达量为7.56±>0.46）,之后逐渐下降,术后21 d降至术前水平。实验组各相邻取材时间点两两比较,血清TNF-α mRNA和IL-1β mRNA表达水平差异均有统计学意义(均为P<0.05)。术后1 d、3 d、7 d、14 d,非手术眼房水TNF-α和IL-1β蛋白浓度均较基线值升高,其中峰值出现于术后7 d［TNF-α蛋白浓度为（162.34±5.71） ng·L^-1,IL-1β蛋白浓度为（16.68±0.74）ng·L^-1］,之后逐渐下降,术后21 d降至术前水平。实验组相邻取材时间点两两比较,非手术眼房水TNF-α和IL-1β蛋白浓度差异均有统计学意义(均为P<0.05)。术后1 d、3 d、7 d、14 d,非手术眼角膜知觉敏感度较基线值升高,于术后7 d角膜知觉敏感度最高;之后角膜知觉敏感度逐渐下降,术后21 d基本恢复至基线水平。实验组非手术眼相邻时间点两两比较,角膜知觉敏感度差异均有统计学意义（均为P<0.05）。结论　实验兔第一眼晶状体超声乳化吸出术后早期,非手术眼角膜知觉敏感度升高,该变化可能与该眼房水及血清中TNF-α和IL-1β表达的动态变化相关。     

晶状体全脱位的人工晶状体植入术

目的观察晶状体全脱位患者行人工晶状体植入的效果与并发症。方法毫针固定脱位晶状体并圈套娩出,显微剪剪除脱出玻璃体加睫状沟缝线固定人工晶状体。术后随访６～１３ｍｏ,平均８．２ｍｏ．结果观察７例７眼术后视力０．８者１眼,０．７者１眼,０．５者３眼,０．３者２眼。术中术后并发症少。结论晶状体全脱位患者经过人工晶状体植入术后,均获得满意视力。     

晶状体或人工晶体脱入玻璃体腔的手术处理

目的探讨晶体或人工晶体脱位入玻璃体腔的处理方法。方法钝挫伤后晶体脱位3眼,白内障手术中或术后晶体或人工晶体脱位11眼。均采用标准睫状体扁平部三切口闭合式玻璃体切除方式。术中注入全氟化碳液体0.4～4ml。经角膜缘取出人工晶体或晶体,或是直接玻璃体腔内粉碎晶体。再植入前房型人工晶体,或行后房型人工晶体睫状沟缝线固定。结果14眼均顺利取出或粉碎玻璃体腔内晶体或人工晶体。13眼(92.86%)术中植入人工晶体。术后视力明显提高12眼(85.71%)。结论对晶体或人工晶体全脱位进入玻璃体腔者,需争取早期采用玻璃体切除联合脱位晶体或人工晶体取出或粉碎手术,争取再植入人工晶体。     

Intraocular lens power calculation for lens exchange

PURPOSE: To examine patients who had intraocular lens (IOL) exchange for large postoperative refractive errors and determine the factors that contributed to the error in IOL power calculation. SETTING: Thirteen affiliated hospitals in Japan. METHODS: This study comprised 34 cases that required IOL exchange because of large refractive errors after primary lens implantation. Patients with intraoperative complications were excluded from the study. The potential contribution of axial length, corneal refractive power, IOL manufacturer, and IOL fixation to errors in the predicted power was examined retrospectively. Axial length was calculated by the SRK/T and Holladay formulas using refraction after primary IOL implantation. RESULTS: There was no statistical difference between the corneal refractive power before and after cataract surgery. The axial lengths calculated using the SRK/T and Holladay formulas were longer than the ultrasonic axial lengths in 24 and 23 cases, respectively. Using IOLs from the same manufacturer for both primary implantation and exchange reduced the error in predicted refraction. CONCLUSION: Axial length and IOL manufacturer were important factors in predicting refraction power in eyes requiring IOL exchange.     

Shape of lens epithelial cells after intraocular lens implantation

PURPOSE: To evaluate the morphological behavior of lens epithelial cells (LECs) after human cataract surgery with implantation of a poly(methyl methacrylate) (PMMA) or silicone intraocular lens (IOL). SETTING: Department of Ophthalmology, Fujita Health University, Banbuntane Hotokukai Hospital, Nagoya, Aichi, Japan. METHODS: Morphological observations of LECs in the patients with IOLs were made by light and transmission electron microscopy. The LECs were from 4 areas: (1) the region below the anterior capsule, touching the IOL; (2) the area between region 1 and the equatorial region; (3) the equatorial region; and (4) the central equatorial region and of the posterior capsule not touching the IOL. Case 1 had implantation of a single-piece IOL with a PMMA optic and haptics. Case 2 had a 3-piece IOL with a PMMA optic and polypropylene haptics. Case 3 had a 3-piece IOL with a silicone optic and polypropylene haptics. Areas 1 and 4 could not be observed in Case 2. RESULTS: The major difference between the patient with a PMMA IOL (Case 1) and the patient with a silicone IOL (Case 3) was that among the 4 areas observed, collagen fibers were present only in area 1 in Case 1 but in areas 2 or 3 as well in Case 3. CONCLUSIONS: Fibrous collagen fibers appeared in regions in which LECs adhered and there was capsule contact with the IOL optic. In addition fibrous collagen fibers appeared in more areas in the eye with the silicone IOL than in that with the PMMA IOL, perhaps because IOLs with silicone optics move slightly while in the capsular bag.     

Patterns of lens care practices and lens product contamination in contact lens associated microbial keratitis

A prospective study of 24 contact lens patients with culture- or histopathology-proven microbial keratitis was performed between July and December 1987 to determine associated lens care practices and patterns of lens care product contamination. A questionnaire was used to document methods of lens care. Bacterial cultures were performed on all available contact lenses, lens cases, and solutions (including saline, disinfectant, daily cleaner and wetting agents), in addition to standard smears and cultures of the corneal ulcer. Failure to follow standard recommendations regarding contact lens care was widespread (21 of 24, 88%). In the majority of patients (20 of 24, 83%) bacterial contamination of the contact lens, case, and/or solutions was present. Almost two-thirds (15 of 24, 62%) of patients used solutions that were more than 3 months old. Cosmetic extended wear lens patients were most likely (7 of 8, 88%) to use solutions that were more than 3 months old and very likely (6 of 8, 85%) to have contaminated solutions. Efforts to improve lens care and decrease lens product contamination are necessary to prevent contact lens associated corneal ulcers.     

Regulation of lens volume: Implications for lens transparency

Lens transparency is critically dependent on the maintenance of an ordered tissue architecture, and disruption of this order leads to light scatter and eventually lens cataract. Hence the volume of the fiber cells that make up the bulk of the lens needs to be tightly regulated if lens transparency is to be preserved. While it has long been appreciated that the lens can regulate its volume when placed in anisosmotic solutions, recent work suggests that the lens also actively maintains its volume under steady-state conditions. Furthermore, the process of fiber cell elongation necessitates that differentiating fiber cells dramatically increase their volume in response to growth factors. The cellular transport mechanisms that mediate the regulation of fiber cell volume in the lens cortex are only just beginning to be elucidated. In this region, fiber cells are continuously undergoing a process of differentiation that creates an inherent gradient of cells at different stages of elongation. These cells express different complements of transport proteins involved in volume regulation. In addition, transport processes at different depths into the lens are differentially influenced by electrochemical gradients that alter with distance into the lens. Taken together, our work suggests that the lens has spatially distinct ion influx and efflux pathways that interact to control its steady-state volume, its response to hypotonic swelling, and the elongation of differentiating fibers. Based on this work, we present a model which may explain the unique damage phenotype observed in diabetic cataract, in terms of the uncoupling or dysregulation of these ion influx and efflux pathways.     

非球面人工晶状体与球面人工晶状体的比较

随着波前像差技术的发展,人们对影响白内障术后视功能的因素有了更深刻的认识,设计出旨在降低人工晶状体眼球面像差从而提高视功能的非球面人工晶状体。现就非球面人工晶状体眼与球面人工晶状体眼相比,在视功能、波前像差等方面有何差异,非球面晶状体的优越性,以及影响优越性表现的因素综述如下。     

晶体脱位处理联合人工晶体植入

总结３７例（３８只眼）处理晶体脱位并同期行人工晶体植入的经验。本文的病例晶体脱位各异，以外为主，包括半脱位及全脱位于前房、玻璃体及视网前，处理方法均采用手术治疗，采用不同的术式去除脱位晶体，同时入前房型或后房型人工晶体。结果表明，治疗后的力均有没程度提高，且并发症少，提示手术效果令人满意。本文讨论了去除晶体的各种术式和植入人工晶体的类型及方法的选择经验。