高糖刺激人视网膜色素上皮细胞诱导型一氧化氮合酶表达及与p38 MAPK信号通路关系

目的:观察高糖刺激体外培养的人视网膜色素上皮(human retinal pigment epithelium,RPE)细胞表达诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)的作用及其与p38信号通路(p38 mitogen activated protein kinase,p38 MAPK)的关系,从而进一步探讨糖尿病视网膜病变的发病机制.方法:培养人RPE细胞,分为对照组、高糖组、高糖 SB203580组和甘露醇组,采用四甲基氮唑蓝(MTT)比色法检测细胞活力,免疫荧光染色法观察RPE细胞中磷酸化p38 MAPK和iNOS的蛋白表达.结果:①MTT检测结果:高糖可以抑制RPE细胞增殖,加入特异性p38 MAPK抑制剂SB203580预处理后,抑制作用减弱;②免疫荧光法检测结果:与对照组相比,高糖组磷酸化p38 MAPK,iNOS蛋白表达明显增高;加入SB203580预处理后,iNOS表达被显著抑制.结论:高糖能够抑制RPE细胞增殖,通过活化p38 MAPK信号通路刺激RPE细胞iNOS的表达,在糖尿病视网膜病变的发生发展中起重要作用.     

实验性糖尿病小鼠视网膜电图振荡电位的变化

目的:探讨高血糖对糖尿病小鼠视网膜电图振荡电位(OP)的影响。方法:C57BL/6雄性小鼠80只分为正常组10只和糖尿病组70只。糖尿病组用STZ造模,4wk后将符合条件的小鼠纳入观察组。检测4,8,12,16,20wk时小鼠OP的表现。结果:在STZ诱导造模后,糖尿病组小鼠与对照组相比体质量增长明显降低,差异有明显统计学意义(P<0.05);建模后糖尿病组血糖明显高于对照组,差异有统计学意义(P<0.01);建模后4wk起,糖尿病小鼠视网膜电图OP1,OP2,OPs振幅有下降趋势,峰潜时延长,与对照组小鼠相比差有统计学意义(P<0.01);20wk时糖尿病小鼠OP1,OP2,OPs振幅降低,峰潜时延迟,与4wk时比较差异有统计学意义(P<0.01)。结论:高血糖可使糖尿病小鼠OP振幅下降,峰潜时延长;且随病程的延长其影响逐渐增强。     

非穿透性小梁切除术与小梁切除术后角膜内皮细胞变化的对比分析

目的探讨角膜内皮细胞在非穿透性小梁切除术与小梁切除术后的变化及临床意义。方法采用非接触眼压计、角膜内皮计及细胞图像分析系统观察14例(16只眼)非穿透小梁切除术与24例(25只眼)小梁切除术患者手术前后眼压、中央角膜厚度、角膜内皮细胞情况,并对结果进行统计学分析。结果非穿透性小梁切除组术后眼压显著下降(P<0.05),中央角膜厚度(CCT)、最小细胞面积(MIN)、最大细胞面积(MAX)、平均细胞面积(AVE)、细胞面积变异系数(CV%)和细胞密度(CD)均较术前无显著性差异(P>0.05);小梁切除组术后眼压显著下降(P<0.05),中央角膜厚度(CCT)较术前增加(P<0.05),平均细胞面积(AVE)较术前增加(P<0.05),细胞密度(CD)及六角形细胞百分数(H%)均较术前显著减少(P<0.05)。结论非穿透性小梁切除术和小梁切除术均可有效控制眼压,非穿透性小梁切除术后角膜内皮细胞变化较小,手术安全性高。     

麻黄碱和去氧肾上腺素对全麻病人眼内压的影响

目的 探讨不同浓度麻黄碱和去氧肾上腺素对全麻病人眼内压(IOP)的影响.方法 48例ASA Ⅰ级择期行腹部手术病人,随机分成4组,每组12例.气管插管后1、2组分别静脉输注麻黄碱5mg、10 mg,3、4组分别静脉输注去氧肾上腺素50μg、100μg,记录入室后(T0)、气管插管后5min(T1)、注药后10min(T2)病人IOP、收缩压(SBP)和心率(HR).结果 1、2组给药后T2的IOP较T0及T1下降(P<0.05);3、4组给药后IOP也有下降,但差异无统计学意义(P>0.05).结论 全麻病人使用50～100 μg去氧肾上腺素对IOP无明显影响,而麻黄碱可使IOP下降.     

缺氧时Genistein对人RPE细胞生长作用机制的初步研究

目的:研究缺氧条件下金雀异黄素(genistein,Gen)对培养人视网膜色素上皮细胞(retinalpigmentepitheliumcell,RPE)的生长影响,探讨缺氧时Gen对人RPE细胞生长的作用机制。方法:以不缺氧为对照,建立人RPE细胞的缺氧模型,分别以Gen0、50、100、200μmol/L(组)的药物浓度作用细胞;用甲基噻唑基四唑(methylthiazolyltetrazolium,MTT)测定法测定24h细胞活力(以吸光度A值表示);用RT鄄PCR(反转录-聚合酶链反应)技术检测细胞内RasmRNA表达;用酶联免疫吸附试验技术(ELISA)检测硝基酪氨酸(3鄄Nitrotyrosine,3鄄NT)的产量;用黄嘌呤氧化酶法测定细胞内超氧化物歧化酶(superoxidedismutase,SOD)的活力。结果:与对照组比,缺氧条件下,随着Gen浓度增高,24h3鄄NT的生成量均增加(P<0.05),SOD代偿性增高(P<0.05);与Gen0组比较,Gen50、100、200组Ras基因相应表达升高。Gen在100、200μmol浓度时,24h细胞MTT检测较Gen0A值显著升高(P<0.05),表明Gen的作用下,缺氧RPE细胞出现增殖现象。结论:Gen增强了缺氧24h的RPE细胞内的氧化应激水平,Ras基因mRNA表达增高,细胞增殖,可能同导致3鄄NT生成增多的自由基有关。     

加替沙星和左氧氟沙星治疗细菌性结膜炎的疗效对比研究

Objective To evaluate the different efficacy and safety to the treatment of baeterial conjunctivitis between the fluoroquinolone antibiotics gatifloxacin and levofloxacin.Methods A multicentre,random,double-blind and control trial was performed in six centers including Eye Ear Nose and Throar Hospital of Fudan University,Henan Eye Institute,Eye Hospital Affiliated to Wenzhou Medieal College,the First Affiliated Hospital Of Nanjing Medical University,Xijing Hospital Affiliated to the Fourth Military Medical University,the Second Affiliated Hospital of Xi'an Jiaotong University between August 2006 and October 2007. The levofloxacin was set as the efficient control. Two hundred and thirty-five patients (235 eyes ) that diagnosed as bacterial conjunctivitis were randomly divided into two groups by the method of randomized blocks, the test group (gatifloxacin) had 118 eyes and the control (levofloxacin) group had 117eyes. The drug delivery into conjunctival sac was performed at a 7-day period ( two drops per time, eight times per day at the first two days and two drops per time, four times per day at the following 5 days). All participants were given the conjunetival sac germ culture and drug sensitive test before and after the study.The combination score of signs and symptoms and evaluation of safety were conducted at the pre-dilivery day,the (4 ± 1 ) and (7 ± 1 ) dilivery day. The statistic analysis was conducted by CMH X2 test, Pearson X2 test and Fisher's exact probabilities test. Results The efficacy of the two groups was 94. 0% ( 110/117 eyes)in gatifloxacin group and 93.8% ( 106/113 eyes) in levofloxacin group with no significant difference( X2 =0.052,P = 0.8201 ). There was also no difference in the bacteria clearance between the two groups [ gatifloxacin versus levofloxacin, 94.1% (80/85 eyes) versus 92. 5% (74/80 eyes) ,P =0. 3470]. The decrease of combination score of signs and symptoms at the (4 ± 1 ) delivery day was 4. 436 +2. 310 in the gatifloxacin group and 3. 814 ± 1. 962 in the levofloxacin group, the difference of which was significant ( F =7.280,P = 0. 0075 ) . This trend was also proved at the ( 7 ± 1 ) delivery day ( gatifloxacin versus levofloxacin, 7. 487 ± 2. 821 versus 6. 912 ± 2. 911, F = 4. 060, P = 0. 0452 ). The visual acuity and the tolerance after local application of the eye drops between the two groups had no diffenrence ( the visual acuity F = 1.04, P = 0. 3080; the tolarence after local admission X2 = 0. 1372, P = 0. 7111 ). According to the result of the germ culture, the major pathogenic bacteria were Gram-positive bacteria ( totally 20 kinds of Grampositive bacteria and 8 kinds of Gram-negetive bacteria). The MIC and drug resistence of gatifloxacin to the Gram-positive bacteria was lower than that of the levofloxaxin ( Staphylococcus Epidermidis, Staphylococcus Aureus, coagulase negative Staphylococcus, α-hemolytic Streptococcus). Conclusions The gatifloxacin eye drop has a good therapeutic effect to the bacterial conjunctivitis, It can effectively clear the pathogen with fast and strong effect. Moreover, it has a low MIC in vitro, advance a prospect in drug resistance, safety and ocular tolerance.     

不典型中心性渗出性脉络膜视网膜病变的临床分析

目的 探讨不典型中心性渗出性脉络膜视网膜病变（中渗）的临床特征。方法 回顾性分析480例自1999年至今临床诊为中心性浆液性脉络膜视网膜病变患者（中浆）的临床资料,并复习其荧光眼底血管造影照片。结果 11例（2．3％）原临床诊为中浆病例者经荧光素眼底血管造影（fundus fluorescein angiography,FFA）检查证实为中渗。所有病例视力均较好（≥0．5）,病变均位于黄斑中心凹外或近黄斑中心凹,均没有出血,呈不典型眼底表现。年龄17～39岁,平均27．5岁。男性多见。结论 不典型中渗易误诊为中浆,FFA有助于两者的诊断与鉴别诊断。     

金雀异黄素对氯化钴诱导的兔视网膜色素上皮细胞血管内皮生长因子表达的影响

目的：研究氯化钴对体外培养的兔视网膜色素上皮(RPE)细胞血管内皮生长因子(vEGF)表达的诱导作用，以及金雀异黄素(genistein，Gen)对其表达的影响。方法：采用免疫荧光组织化学技术和激光共聚焦显微镜，以荧光比值来表示VEGF的表达，半定量分析氯化钴处理不同时间对体外培养的BPE细胞VEGF蛋白表达的诱导作用，以及Gen对低氧或氯化钴诱导的VEGF表达的影响。结果：对照组有低水平VEGF蛋白的表达，随着低氧或氯化钴处理时间的延长，荧光比值的变化表现为时间依赖性，在6h点达到高峰。Gen(50、100、200μmol/L组)与二甲基亚砜(DMSO)对照组相比，均可明显抑制氯化钴诱导的VEGF的表达，且此种抑制作用呈浓度依赖性。结论：Gen可抑制氯化钴诱导的RPE细胞VEGF表达，提示Gen对视网膜新生血管的形成具有潜在的预防及治疗价值。     

TGF-β1诱导人Tenon囊成纤维细胞增殖及其机制

目的:观察TGF-β1是否促进人Tenon囊成纤维细胞(HTFs)增殖并且探讨其作用机制?方法:用TGF-β1(0?0.5?1.0?2.0?5.0?10.0 ng/ml)作用于体外培养的3~6代HTFs 24 h,CCK-8 法?EdU染色法检测其增殖;采用Western blot法检测HTFs中RhoE的表达及其下游细胞增殖相关蛋白cyclinD1和增殖细胞核抗原(PCNA)的表达,用慢病毒建立RhoE敲降细胞系?结果:TGF-β1可以剂量依赖性促进HTFs增殖,以5 ng/ml TGF-β1处理后有统计学差异,EdU掺入法检测发现,5 ng/ml TGF-β1组EdU标记率明显高于对照组?5 ng/ml TGF-β1组 HTFs中RhoE蛋白的表达平均灰度值明显低于对照组,细胞增殖相关蛋白cyclinD1?PCNA的表达平均灰度值高于对照组?细胞中RhoE沉默后,CCK-8法检测细胞活力较对照组上升,流式细胞术发现S期也升高,cyclinD1?PCNA灰度值也上升?结论:TGF-β1可以通过降低RhoE引起下游周期蛋白cyclinD1的表达升高,诱导人Tenon囊成纤维细胞增殖?     

金雀异黄素抑制缺氧人视网膜色素上皮细胞VEGF表达机制的初步研究

目的:进一步明确金雀异黄素(Gen)抑制缺氧人视网膜色素上皮细胞(RPE)血管内皮生长因子(VEGF)表达的机制。方法:以体外正常情况为对照,建立人RPE缺氧模型并经不同浓度Gen干预后,通过RT-PCR检测VEGF及其转录调节因子c-fos、c-jun、缺氧诱导因子(HIF)-1α、HIF-2α、HIF-3αmRNA的表达水平,了解Gen是否通过对VEGF的转录因子活性调节而影响VEGF的表达。结果:缺氧能使人RPE细胞的VEGF、c-fos、c-jun及HIF-2α、HIF-3αmRNA水平增高,对HIF-1α无影响;Gen能抑制缺氧诱导的VEGF表达;在所检测的VEGF转录因子中,Gen只对HIF-2α的转录有抑制作用。结论:Gen抑制VEGF表达的作用部分是通过抑制其转录调节因子HIF-2α转录实现的。