γ干扰素对兔晶状体上皮细胞增殖细胞核抗原表达的影响

目的 :探讨γ干扰素 (interferon γ ,IFN γ)对兔晶状体上皮细胞增殖细胞核抗原 (proliferatingcellnuclearcntigen ,PCNA)表达的影响。方法 :采用新西兰白兔白内障囊外摘除模型 ,术中囊袋内灌注BSS为A组即对照组。B、C组为治疗组 ,分别灌注 10 3 IU/mlIFN γ和 10 4IU/mlIFN γ。术后 3个月运用免疫组化法和计算机图象分析技术检测晶状体上皮细胞PCNA表达。结果 :γ干扰素组PCNA阳性表达少 ,与对照组比较有显著性差异 (P <0 0 5 ) ,高浓度抑制作用强。结论 :IFN γ有抑制兔晶状体上皮细胞PCNA表达的作用 ,IFN γ对PCNA表达的抑制作用具有浓度依赖性     

视网膜母细胞瘤增殖细胞核抗原表达的研究

目的 :探讨增殖细胞核抗原 (proliferatingcellnuclearantigen ,PCNA)在视网膜母细胞瘤 (retinoblastoma ,Rb)中的表达及其与Rb病理类型和侵犯视神经情况的关系。方法 :用免疫组化技术检测PCNA在 3 7例Rb组织标本中的表达 ,并应用多功能真彩色病理图像分析系统 (CMIAS系列 )来分析PCNA染色的阳性细胞个数 ,采用积分光密度值定量分析其增殖指数 ,并将各指标进行SPSS统计软件相关性分析。结果 :本组 3 7例Rb标本中PCNA的阳性表达为 97 3 0 % ,而在 2例正常视网膜组织内 (对照组 )的表达均为阴性。PCNA的表达水平与Rb不同的分化、增殖程度有显著的统计学差异 (P <0 0 1) ,Rb侵犯视神经情况越严重、分化程度越差 ,PCNA的表达越强。结论 :Rb中PCNA的高表达可作为判断Rb恶性及其程度的新型标记物。PCNA与Rb分化类型、视神经侵犯情况密切相关 ,为PCNA表达水平反映Rb分化程度和增殖能力提供理论依据     

比较5-Fu和MMC缓释膜在青光眼滤过手术中应用的实验研究

目的 评估生物缓释膜在青光眼滤过手术中的安全性和作用,比较5-氟尿嘧啶(5-Fu)和丝裂霉素(MMC)缓释膜在青光眼滤过手术中的抗增殖的疗效.方法 以5-Fu和MMC作为模型药物,壳聚糖作为载体,溶剂挥发法成膜;通过共价交联的方式将12μg的5-Fu或MMC结合到生物缓释膜上.未植入缓释膜作为对照组,其余3组植入空白、5-Fu和MMC缓释膜.四组均行青光眼滤过手术,静脉留置针作为引流管,空白缓释膜、5-Fu缓释膜、MMC缓释膜缝于巩膜瓣下.滤过手术前和手术后1、3、5、714、21和28d,以Tonopen眼压计记录兔眼压,以裂隙灯观察滤过泡大小和眼前节的变化并照相;分别于术后28d每组处死两只兔,总共8只眼标本行病理组织学检查;用扫描电子显微镜检测每组术后28d的角膜和晶状体标本.结果 术前各组平均眼压无差异,MMC组手术前和手术后28d内比较差异有统计学意义(F值为26.866 P＜0.01),5-Fu组和单纯引流管组手术前和手术后14d内比较差异有统计学意义,(F值分别为13.467,6.567 P＜0.01),空白缓释膜组手术前和手术后7d内比较差异有统计学意义(F值分别为11.426 P＜0.01);MMC、5-Fu组滤过泡生存时间优于单纯引流管组、空白缓释膜组.5-Fu和MMC组,术后28d角膜内皮细胞和晶状体前囊无异常改变.结论 5-Fu和MMC生物缓释膜能明显提高滤过手术成功率并且是安全的,在降低眼内压和延长滤过泡减少眼前节并发症方面,MMC缓释膜比5-Fu缓释膜能更有效地提高滤过手术成功率.     

丝裂霉素C对准分子激光屈光性角膜切削术后角膜混浊的实验研究

目的探讨丝裂霉素Ｃ对准分子激光屈光性角膜切削术后角膜上皮下雾状混浊（ｈａｚｅ）的影响。方法对４５只新西兰白兔的双眼行准分子激光屈光性角膜切削术,术中、术后分别给予０．００８％丝裂霉素Ｃ、０．１％地塞米松治疗及空白对照。术后分别进行裂隙灯、角膜内皮镜、光镜和透射电镜检查。结果术后４及８周,丝裂霉素Ｃ组角膜ｈａｚｅ轻于对照组和地塞米松组;地塞米松组轻于对照组。术后１、４及８周,丝裂霉素Ｃ组术区前基质内角膜细胞数较对照组和地塞米松组少,后两组间差异不明显。三组间角膜上皮愈合时间、角膜上皮厚度及内皮细胞密度差异无显著性（Ｐ＞０．０５）。结论丝裂霉素Ｃ能通过抑制角膜细胞的生长而减轻ｈａｚｅ的形成,其效果优于地塞米松,且无明显副作用,是一种较理想的抑制ｈａｚｅ形成的药物     

Upper eyelid retraction after glaucoma filtering surgery and topical application of mitomycin C

PURPOSE: To describe 3 cases of upper eyelid retraction after glaucoma filtering surgery and topical application of mitomycin C and to highlight possible causes. METHODS: A report of 3 patients, identified over an 8-year interval period, who had development of upper eyelid retraction a few months after undergoing trabeculectomy with mitomycin C in the ipsilateral eye. RESULTS: Testing for Graves disease, including second-generation thyrotropin receptor antibodies (TRAb), was negative in all cases. A complete physical examination was performed by an internist: Orbital and intracranial neuroimaging studies were ordered when necessary, and all were negative. One patient underwent surgical repair of the eyelid retraction and had a satisfactory result. CONCLUSIONS: Upper eyelid retraction after filtering surgery is a rarely reported entity. Müller muscle overaction, independent of Graves disease, is a likely cause and has been pointed out as a possible factor in one previous report. Müller muscle fibrosis, euthyroid Graves disease, and mechanical hindrance to an elevated bleb are other possible factors.     

滤过手术中丝裂霉素C不同用法的疗效分析

目的 比较青光眼滤过手术中丝裂霉素C（MMC）结膜瓣下与巩膜瓣下两种应用方法的降压效果和并发症的发生情况。方法 132眼接受滤过手术的原发性青光眼随机分为对照组和MMC组，再将应用MMC（0．4mg／ml，3min）的66眼分成结膜瓣下（MMC-A）组和巩膜瓣下（MMC-B）组。观察1年内3组患者手术后的眼压、视力和手术并发症。结果 MMC组与对照组相比眼压下降差异有统计学意义（P〈0．05），功能性滤过泡形成率71．21％，手术成功率达75．76％，而两组手术并发症无显著性差异。MMC两组病人的降压效果和并发症的差异无统计学意义。结论 MMC作为滤过性手术辅助药物，可增加手术成功率，而且无严重并发症。MMC放置在结膜瓣下和巩膜瓣下效果相似，但放置在结膜瓣下可能会减少对眼内组织的毒性。     

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。     

曲古抑菌素A对兔眼滤过术后滤过泡作用的研究

目的：观察曲古抑菌素A( trichostatin A,TSA)作用于兔眼滤过术后滤过泡的形态变化,研究其对术后结膜瘢痕的抑制作用。
　　方法：兔眼滤过术中结膜下注射TSA、丝裂霉素C( MMC)、PBS,分别于术后3,7,14,21,28d应用Krofeld评分评价滤过泡的形态变化。
　　结果：TSA组14 d内滤过泡弥漫性隆起,28 d囊性泡形成。术后14,21 d TSA组滤过泡评分高于PBS组,具有统计学差异(P<0.05)。
　　结论：TSA能够抑制术后结膜瘢痕形成,延长滤过泡存在时间,保持滤过道通畅。     

视网膜色素变性Rd1小鼠眼球中增殖细胞核抗原（PCNA）的表达

目的　探讨增殖细胞核抗原（proliferating cell nuclear antigen,PCNA）在Rd1小鼠视网膜上的表达情况。方法　取出生后14 d、21 d、28 d的Rd1小鼠和C57BL/6J小鼠各5只,摘出眼球进行HE染色,观察视网膜形态学结构变化;免疫组织化学染色与实时荧光定量PCR检测PCNA蛋白和基因在不同鼠龄小鼠中的表达。结果　HE染色结果显示:PCNA蛋白均在两种小鼠视网膜神经节细胞中表达;C57BL/6J小鼠视网膜结构完整;与C57BL/6J小鼠相比,Rd1小鼠视网膜发育随着鼠龄的增加,发生进行性退化。免疫组织化学染色结果显示:PCNA蛋白表达仅局限于视网膜神经节细胞,免疫阳性信号出现在第14天和第21天,而在第28天未检测到。PCR 实验结果显示:与C57BL/6J小鼠相比较,鼠龄21 d时Rd1小鼠PCNA基因表达水平增高,是C57BL/6J小鼠的2.23倍;鼠龄14 d和28 d时 Rd1小鼠PCNA基因表达均低于C57BL/6J小鼠（均为P<0.05）。结论　PCNA基因参与了Rd1小鼠视网膜发育退化过程。     

青光眼濾過術中絲裂霉素用量控制方法的探討

目的比較研究Schirmer濾紙片和棉球作爲載體在青光眼手術中對絲裂霉素C應用量的控制.方法將棉球(4.5mm)和Schirmer濾紙(4.5mm)分别浸入濃度爲0.4mg/ml的絲裂霉素溶液中,然後置于以穹窿部爲基底的結膜瓣下5分鍾,并應用可鬆解縫綫.結果含絲裂霉素C濾紙的降低了手術并發癥,有顯著性差别(P<0.1).結論在Schirmer濾紙中的絲裂霉素C含量較易控制,此方法安全且易應用,而棉球中絲裂霉素C含量難控制且增加了手術并發癥.     

5—氟尿嘧啶与丝裂霉素C在青光眼滤过术中的合理应用

目的 探讨青光眼滤过手术抗代谢药物丝裂霉素C（MCC）与5－氟尿嘧啶（5－Fu）的合理应用。方法 对108只需手术的青光眼随机分成3组。5－Fu组术后球结下注射5－Fu6－8次,MMC组术中一次性给予质量浓度为0.2mg/ml的MMC,对照组则不用任何药物,观察各组术后的疗效及并发症。结果 随访3－34个月（平均28．2月）,5－Fu和MMC组手术成功率明显高于对照组,P＜0．05。术后并发症5－Fu组角膜上皮点状损害18只眼（45％）,MMC组低眼压2例（6．7％）,与对照组比有显著差异（P＜0．05）。结论 5－Fu与MMC能提高青光眼手术的成功率,但有一定的毒副作用。临床上应根据病人的年龄、病情及青光眼类型选择用药。     

增殖细胞核抗原在人眼小梁细胞增殖研究中的应用

目的 探讨增殖细胞核抗原（ｐｒｏｌｉｆｅｒａｔｉｎｇ ｃｅｌｌ ｎｕｃｌｅａｒ ａｎｔｉｇｅｎ,ＰＣＮＡ）在细胞增殖研究中的应用价值。方法 利用免疫组化方法对不同生长期（１２小时至２０天）的人眼第四代小梁细胞ＰＣＮＡ表达状况进行研究,并观察在不同浓度肾上腺素、地塞米松及表皮生长因子作用下ＰＣＮＡ的表达水平,并与正常细胞ＰＣＮＡ的表达水平对比研究,借助计算机图像处理系统对结果进行分析。结果 根据正常     

Effects of intraoperative sponge mitomycin C and 5-fluorouracil on scar formation following strabismus surgery in rabbits

purpose?To investigate the influence of 5-fluorouracil (5-FU) and mitomycin C (MMC) on the postoperative adhesions following strabismus surgery in rabbits. methods?Twenty-one New Zealand white rabbits were used in this prospective, masked, controlled trial. Both eyes of 20 animals underwent 3-mm recession of the superior rectus muscle (SRM). In group I (10 animals), one eye of each animal received topical application of MMC (0.2?mg/ml) for 5 minutes and the other eye (control eye) was treated with balanced salt solution (BSS) using an intraoperative sponge. In group II (10 animals), a randomly chosen eye of each animal was treated with 5-FU soaked sponges (50?mg/ml) for 5 minutes and the fellow eye (control eye) with BSS. Two eyes of a rabbit were included as unoperated controls. Four weeks after the surgery, conjunctival vascularity and postoperative adhesions between the SRM Tenon's capsule (TC) and SRM sclera (scl) were assessed. Additionally, eyes were enucleated and evaluated histopathologically for evidence of scarring, granuloma formation, and muscle tissue changes under a light microscope. results?MMC-treated eyes had a higher rate of avascular conjunctiva compared to both controls and 5-FU-treated eyes. Mean adhesion scores, particularly between the SRM-scl, were lower in eyes treated with antiproliferative agents compared to controls. The difference was statistically significant in MMC-treated eyes for the adhesions between SRM-scl (p = 0.03). Histopathological examination revealed less scarring and granuloma formation in MMC- and 5-FU-treated eyes compared to their control eyes. conclusions?MMC, and to a lesser extent 5-FU, are shown to be effective in reducing postoperative scarring following strabismus surgery in rabbits. It seems reasonable to suggest that antimetabolites should be used for cases having an increased risk of postoperative adhesions.     

Effects of intraoperative sponge mitomycin C and 5-fluorouracil on scar formation following strabismus surgery in rabbits

PURPOSE: To investigate the influence of 5-fluorouracil (5-FU) and mitomycin C (MMC) on the postoperative adhesions following strabismus surgery in rabbits. METHODS: Twenty-one New Zealand white rabbits were used in this prospective, masked, controlled trial. Both eyes of 20 animals underwent 3-mm recession of the superior rectus muscle (SRM). In group I (io animals), one eye of each animal received topical application of MMC (0.2 mg/ml) for 5 minutes and the other eye (control eye) was treated with balanced salt solution (BSS) using an intraoperative sponge. In group II (10 animals), a randomly chosen eye of each animal was treated with 5-FU soaked sponges (50 mg/ml) for 5 minutes and the fellow eye (control eye) with BSS. Two eyes of a rabbit were included as unoperated controls. Four weeks after the surgery, conjunctival vascularity and postoperative adhesions between the SRM Tenon's capsule (TC) and SRM sclera (scl) were assessed. Additionally, eyes were enucleated and evaluated histopathologically for evidence of scarring, granuloma formation, and muscle tissue changes under a light microscope. RESULTS: MMC-treated eyes had a higher rate of avascular conjunctiva compared to both controls and 5-FU-treated eyes. Mean adhesion scores, particularly between the SRM-scl, were lower in eyes treated with antiproliferative agents compared to controls. The difference was statistically significant in MMC-treated eyes for the adhesions between SRM-scl (p = 0.03). Histopathological examination revealed less scarring and granuloma formation in MMC- and 5-FU-treated eyes compared to their control eyes. CONCLUSIONS: MMC, and to a lesser extent 5-FU, are shown to be effective in reducing postoperative scarring following strabismus surgery in rabbits. It seems reasonable to suggest that antimetabolites should be used for cases having an increased risk of postoperative adhesions.     

实验性视网膜脱离及复位后视网膜色素上皮细胞抗增殖性细胞核抗原的表达

目的 观察视网膜脱离和复位过程中抗增殖性细胞核抗原(proiferating cell nuclear antigen, PCNA)表达的状况，以评价其在接触抑制机制中的意义。 方法 对72只猫眼行晶状体 囊外摘除和玻璃体切除手术。3周后，利用微穿刺技术制作视网膜脱离和复位动物模型，在不同时间取眼球并制成组织切片。采用免疫组织化学方法，观察视网膜脱离后视网膜色素上皮(retinal pigment-epithelium, RPE)细胞PCNA的表达。光镜下确定PCNA阳性的RPE细胞并量化增殖反应。采用方差分析(ANOVA)方法 ，对脱离组的脱离区和非脱离区及复位组复位区RPE细胞层PCNA的表达水平进行比较。 结果 在视网膜脱离后24 h，脱离组脱离区视网膜RPE细胞层出现PCNA阳性细胞 ，5～6 d达到高峰，20 d后逐渐回落。在复位组复位区，PCNA标记的RPE细胞明显少于脱离组脱离区。脱离组非脱离区视网膜很少有PCNA标记的RPE细胞。3组PCNA标记的RPE细胞数比较，差异有显著性的意义。 结论 视网膜脱离诱导RPE细胞增殖，而视网膜复位后这种增殖受到抑制。表明视网膜复位过程中，接触抑制的机制在发挥作用。（中华眼底病杂志，2003，19：20-23）     

喉非典型增生中PCNA、P21的表达

目的探讨PCNA、P21的表达与喉非典型增生的预后是否相关.方法用免疫组化ABC法对63例喉非典型增生(其中40例转化为癌,另23例未转化)、56例喉鳞癌和7例正常喉组织进行了PCNA、P21检测.结果PCNA指数和P21阳性率在非典型增生转化组与癌间、未转化组与癌间差别均有统计意义(p＜0.01),在非典型增生转化组与未转化组之间差别无统计意义.结论PCNA对衡量喉非典型增生及喉癌的细胞增殖状态有一定的价值,P21与喉癌的发生机制有关,但两者的表达均与喉非典型增生的预后无明显关系.     

Macular hole and subhyaloid hemorrhage following filtering surgery with mitomycin C

Background We report the occurrence of a macular hole with subhyaloid hemorrhage following filtering surgery with mitomycin C.Methods Combined trabeculotomy and trabeculectomy with mitomycin C was performed in the left eye for primary congenital glaucoma.Results Two weeks postoperatively the fundus showed a macular hole with subretinal and retinal hemorrhage along with subhyaloid bleeding. Optical coherence tomography confirmed the presence of a full-thickness macular hole.Conclusion A full-thickness macular hole with subretinal and retinal hemorrhage is reported as a complication of glaucoma filtering surgery.     

Ribozyme to proliferating cell nuclear antigen to treat proliferative vitreoretinopathy

PURPOSE: A DNA-RNA chimeric ribozyme was developed that targets the mRNA of a cell cycle regulatory protein, proliferating cell nuclear antigen (PCNA). The hypothesis was that inhibition of PCNA, essential in DNA replication, would decrease the proliferation of cells that are involved in formation of granuloma after surgical procedures in the eye. The ability of intravitreous injection of this ribozyme to prevent or inhibit development of proliferative vitreoretinopathy (PVR) was tested in a dispase-induced rabbit PVR model. METHODS: Rabbit genomic DNA encoding PCNA was cloned and sequenced. The cleavage of rabbit PCNA by the chimeric ribozyme was tested in vitro. Delivery of the ribozyme to rabbit retinal pigment epithelial (RPE) or fibroblast cells and its effects on proliferation of fibroblasts were examined. The stability of the ribozyme in vitreous fluid and serum was studied as well. In the dispase-induced rabbit model of PVR, the ability of the PCNA ribozyme to prevent or inhibit development of PVR and retinal detachment (RD) was tested. Experimental groups receiving intravitreous PCNA ribozyme, with or without a lipid vehicle, were compared with sham-treated control groups. Progression of PVR in rabbit eyes was followed by indirect ophthalmic examination and observations documented by fundoscopic photography, gross pathology, and histopathology. RESULTS: The chimeric ribozyme targeted a specific sequence in the rabbit PCNA that was identical with that in the human. In vitro cleavage assays confirmed the ability of the ribozyme to cleave the mRNA of PCNA. The catalytic efficiency in vitro, calculated as k(2)/K(m)(app), was 0.26 microM(-1) x min(-1). In vitro studies with fluoresceinated ribozyme indicated that lipid vehicles facilitated delivery of the ribozyme into cells causative of PVR (RPE and fibroblasts); however, the PCNA ribozyme decreased the proliferation of fibroblasts, with or without lipid vehicle. The ribozyme displayed good stability in vitreous fluid, whereas, it degraded quite rapidly in serum. In animal experiments, rabbits in sham-treated groups usually exhibited development of severe PVR characterized by focal traction or RD. Animals in the PCNA ribozyme-treated groups usually did not exhibit an RD. If they did have RD, it was small and localized, or focal tractions developed that did not progress to the degree that the sham-treated animal eyes did over the follow-up period. The in vivo use of a lipid delivery vehicle resulted in a precipitate; however, an effective naked ribozyme dose was identified that did not cause this side effect. CONCLUSIONS: In addition to validating the newly developed dispase PVR rabbit model, the results indicate that ribozyme targeted against the cell cycle agent PCNA is efficacious in the treatment or prevention of PVR in the rabbit eye. These experiments suggest that chimeric ribozyme targeted against PCNA may have a therapeutic or preventative role in humans.