表皮生长因子对兔眼角膜修复作用的量效关系研究

用角膜表皮刮磨方法人工造成兔眼角膜创伤直径为10mm的圆形创面，分析小鼠表皮生长因子（EGF）对其修复的剂量关系。5个浓度EGF（100、50、20、10和0．5μm／ml）治疗的试验组痊愈时间均比对照组提前，有非常显著的差异（P＜0.01），但在这5个浓度组之间，痊愈时间无显著性差异。结果表明，小鼠EGF对兔眼角膜损伤有促修复作用，实验所用的5个浓度与修复作用无剂量依赖关系。     

胚胎干细胞向体表外胚层及角膜上皮细胞分化过程中信号通路的调节

干细胞泛指体外具有自我更新和多向或定向分化潜能的细胞,胚胎干细胞则可被诱导分化形成内胚层、中胚层及外胚层来源的200余种不同细胞。角膜上皮细胞来源于胚胎发育时期的表皮外胚层。临床上严重、广泛的眼表损伤,需要具有增殖能力的角膜上皮细胞修复损伤。利用体内外研究体系模拟体内胚胎发育及组织环境,阐明具有生物活性的角膜上皮细胞的发育过程,为再生医学提供依据,是近年来研究的热点之一。本文介绍了Notch、Wnt、骨形态发生蛋白及成纤维生长因子等调控表皮外胚层发育及与角膜上皮细胞分化相关的信号通路,以既往实验研究为基础,就各信号通路调控的机制及主要调控因子进行综述。     

重组人表皮生长因子对冷冻角膜缘上皮细胞PCNA表达的影响

为探讨重组人表皮生长因子(rhEGF)的浓度对冷冻后兔角膜缘上皮细胞增殖活性的影响，采用器官培养方法和SABC免疫组织化学染色法检测角膜缘上皮细胞的PCNA表达。结果表明，器官培养1周时，冷冻角膜缘上皮细胞核中均可见PCNA表达，染色阳性细胞主要在基底细胞层。EGF组的PCNA染色阳性率显著高于无EGF组(P&;lt;0．01)；而不同浓度的EGF组之间阳性率差异无显著意义(P&;gt;0．05)。结论：在体外器官培养中，EGF能增强超低温冷冻保存的角膜缘上皮细胞PCNA的表达，但无量效依赖性。     

诺沛眼胶治疗芥子气角膜损伤的试验观察

芥子气角膜损伤是一种特殊的化学伤，主要损伤角膜上皮。角膜上皮的再生对角膜损伤的修复至关重要，如果上皮愈合延迟或不能愈合，则可能造成角膜感染甚至穿孔等并发症。因此，加速角膜上皮的愈合一直是临床积极探求的方向。目前尚无特异性治疗药物或措施来阻止芥子气对组织的进行性损伤，而只能对已造成的损伤进行修复及防止继发性损伤。     

干细胞培养角膜上皮移植治疗眼表疾病获得突破

采用体外培养角膜缘干细胞的方式，构建角膜上皮层，移植治疗兔子角膜缘损伤，术后角膜透明，上海第二医科大学附属第九人民医院的这项研究为临床治疗眼表疾病提供了理论依据和实践经验，这一成果日前通过专家鉴定。     

维生素A对糖基化终末产物诱导的角膜上皮细胞损伤的作用及机制研究

目的糖基化终末产物(AGEs)在糖尿病患者在角膜外伤或接受角膜手术时极易出现角膜上皮愈合延迟甚至不愈的过程中起着关键作用,本文研究维生素A(VA)对以糖基化终末产物AGE-BSA诱导角膜上皮细胞损伤的影响及可能的作用机制。方法以HCE-2细胞为研究对象,AGE-BSA为诱导剂,试剂盒检测VA对细胞凋亡率及线粒体膜电位的影响,荧光显微镜检测细胞内凋亡小体,Western blot检测相关的蛋白表达量。结果VA抑制AGE-BSA诱导的HCE细胞凋亡,调节AGEBSA引起的凋亡相关蛋白的变化。同时,VA与JNK的抑制剂同等程度的减少AGE-BSA诱导的JNK和NF-κB的磷酸化。结论VA可能通过抑制JNK介导的NF-κB炎症通路而起到减少AGE-BSA诱导的角膜上皮细胞凋亡的作用,可以作为糖尿病患者角膜上皮损伤的保护剂。     

穿透性角膜移植术围手术期护理

角膜病是眼科的主要致盲眼病，角膜移植是角膜盲的唯一复明方法。角膜移植手术就是用正常透明的异体角膜组织置换病变混浊的角膜组织，达到治疗角膜疾病，使患眼复明的目的。我院已开展角膜移植手术近20年，施行该手术近500例。总结我院2004～2006年施行的穿透性角膜移植术36例的护理体会如下。     

角膜长期保存的研究进展

<正>角膜病是当今世界主要致盲眼病之一。以目前的医疗条件,角膜移植术是使角膜盲患者恢复视力的主要手段。由于受传统观念的影响,在我国能捐献角膜的人数十分有限,角     

32例爆炸致非穿透性角膜外伤临床治疗分析

目的探讨不同损伤程度的爆炸性角膜外伤治疗的临床疗效。方法回顾性分析病例32例爆炸性角膜外伤,8眼角膜裂伤<3 mm者,行角膜浅层异物剔除等保守治疗;30眼角膜裂伤>3 mm且伤口不规整伤口者行角膜异物剔除、角膜裂伤缝合、羊膜遮盖的综合治疗。对角膜上皮修复、角膜裂伤愈合、角膜新生血管生长情况进行观察分析。结果 8眼角膜裂伤<3 mm者术后1周角膜上皮愈合,遗留浅层角膜薄翳。30眼角膜裂伤>3 mm者术后3～4周羊膜溶解后观察到角膜上皮愈合、角膜裂伤瘢痕混浊;随访1～6个月,84.21%(32/38眼)无角膜新生血管生长。结论对不同损伤的爆炸性角膜外伤采用不同治疗方法是切实有效的,羊膜遮盖术在促进角膜上皮愈合、减轻瘢痕混浊和抑制新生血管生长方面疗效显著。     

bFGF促进角膜损伤修复作用及部分机制研究

目的:研究碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)促进兔角膜基质细胞的增殖、迁移作用,探讨bFGF促进兔角膜基质细胞增殖、迁移的分子机制。方法:使用兔角膜基质细胞作为细胞模型,分为对照组、bFGF组、bFGF+ERK1/2或者p38抑制剂组。用四甲基偶氮唑盐比色法(MTT法)检测细胞增殖,细胞划痕实验检测细胞迁移水平,Western blot检测ERK1/2及其磷酸化蛋白和p38及其磷酸化蛋白的表达。结果:MTT结果显示,bFGF对兔角膜基质细胞有明显的促进增殖的作用,当bFGF浓度在2mg·L-1时促增殖作用最为明显;细胞迁移实验结果显示,用2mg·L~(-1) bFGF处理兔角膜基质细胞48h,能够明显地促进该细胞迁移。Western blot实验结果表明,bFGF能够上调和细胞增殖相关的ERK1/2信号通路磷酸化水平以及和细胞迁移相关的p38 MAPK磷酸化水平。当分别加入信号通路抑制剂后,bFGF促兔角膜细胞增殖和迁移作用都受到一定程度的抑制,同时伴随着相关通路的关键蛋白的磷酸化水平降低。结论:bFGF能够激活ERK1/2和p38 MAPK信号通路,从而提高兔角膜基质细胞的增殖、迁移,促进角膜损伤修复。     

中山市横栏镇工人眼外伤（角膜异物）成因分析

目的探讨眼外伤（角膜异物）成因情况。方法采用回顾性分析的方法 ,分析我院2003～2009年眼科收治的角膜异物患者临床资料。结果 2003～2009年角膜异物发病率逐年增高,患者年龄〈30岁为发病集中年龄、夏秋季节为主要时间段、电焊、切割工人为发病主体、异物主要以铁屑和工业粉末为主,异物多侵犯角膜浅层,治疗后患者视力恢复明显,P〈0.05。结论角膜异物是常见的职业性眼外伤,改善工作环境和有效的个人防护能有效的预防角膜异物的发生。     

COMPARATIVE EVALUATION OF SEVERAL METHODS AND CONDITIONS FOR THE IN VIVO MEASUREMENT OF CORNEAL THICKNESS IN RABBITS AND RATS

In vivo corneal thickness (CT) measurements can be used to predict corneal injuring potential in low-volume ocular instillation and low-concentration vapor exposure studies. In this study, CT was measured using several techniques and conditions to assess the most accurate and humane methods for use in predicting corneal injury. Initially, CT was measured in the unanesthetized cornea of unsedated rabbits using an optical or ultrasonic pachymeter; accurate measurements could not be obtained because of rabbit excitability. Subsequently, changes in corneal thickness were evaluated in rabbits following treatment with combinations of the sedative ketamine, the topical anesthetic proparacaine, and/or the mild irritant dichloromethane (DCM; 0.1 mL) using an ultrasonic pachymeter. Mean CT was up to 17% lower in eyes of sedated rabbits receiving proparacaine than in eyes without proparacaine. For eyes tested with DCM and proparacaine, CT increases correlated well with observed eye irritation (r = 0.73). Corneal injury and iritis were more severe in animals without topical anesthesia. An ultrasonic pachymeter was used to measure CT in rabbits previously conditioned to remain calm in cat bags, and in rats restrained with paper towels; no sedative or topical anesthetic was used. After instillation of DCM, CT increased in rabbits corresponding to the severity of irritation (r = 0.79). Untreated eyes showed consistent values throughout the day. CT was also measured in control (untreated) rats and a group of rats having bis\[2-(dimethylamino)ethyl]ether, an irritant, applied to the eye. Changes were measured in both irritant-treated and control eyes. CT values in untreated rat eyes varied by up to 21%, but the corneal thickness of the irritant-treated eyes doubled. The findings demonstrated that a relatively good correlation between corneal thickness and ocular irritation can be obtained using an ultrasonic pachymeter on gently restrained animals. The data confirm that measurement of corneal thickness is a useful and sensitive quantitative method for assessment of eye irritation.     

Clinical progression of ocular injury following arsenical vesicant lewisite exposure

Ocular injury by lewisite (LEW), a potential chemical warfare and terrorist agent, results in edema of eyelids, inflammation, massive corneal necrosis and blindness. To enable screening of effective therapeutics to treat ocular injury from LEW, useful clinically-relevant endpoints are essential. Hence, we designed an efficient exposure system capable of exposing up to six New-Zealand white rabbits at one time, and assessed LEW vapor-induced progression of clinical ocular lesions mainly in the cornea. The right eye of each rabbit was exposed to LEW (0.2?mg/L) vapor for 2.5, 5.0, 7.5 and 10.0?min and clinical progression of injury was observed for 28 days post-exposure (dose–response study), or exposed to same LEW dose for 2.5 and 7.5?min and clinical progression of injury was observed for up to 56 days post-exposure (time–response study); left eye served as an unexposed control. Increasing LEW exposure caused corneal opacity within 6?h post-exposure, which increased up to 3 days, slightly reduced thereafter till 3 weeks, and again increased thereafter. LEW-induced corneal ulceration peaked at 1?day post-exposure and its increase thereafter was observed in phases. LEW exposure induced neovascularization starting at 7 days which peaked at 22–35 days post-exposure, and remained persistent thereafter. In addition, LEW exposure caused corneal thickness, iris redness, and redness and swelling of the conjunctiva. Together, these findings provide clinical sequelae of ocular injury following LEW exposure and for the first time establish clinically-relevant quantitative endpoints, to enable the further identification of histopathological and molecular events involved in LEW-induced ocular injury.     

准分子激光原位角膜磨镶术后角膜瓣移位的处理观察

目的回顾分析准分子激光原位角膜磨镶术（laseriu srtu keratomileusis,IASTK）术后角膜瓣移位的原因,总结出预防这种并发症的方法以及相应的处理措施。方法在使用美国PDI准分子激光治疗仪进行LASIK手术的2613例（5002只眼）中,均应用MoriaⅡ（110刀头）显微角膜刀制作薄角膜瓣。记录到术后发生角膜瓣皱褶、移位患者13例（13只眼）,分析其原因,并给予及时妥善处理,采取相应的治疗对策。结果 LASIK术后第1天出现角膜瓣皱褶、瓣移位8例（8只眼）;术后1w～1y眼外伤致角膜瓣皱褶、移位5例（5眼）。11例（11眼）均行角膜瓣重新复位。术后1个月均达到术前矫正视力。结论 LASIK术中角膜瓣薄、角膜瓣大、蒂部小,易引起角膜瓣皱褶或移位;眼部外伤也是引起LASIK术后角膜瓣皱褶或移位的主要原因之一。如发现及时、治疗得当,仍可达到从前的有效视力。     

透明角膜小切口白内障超声乳化术后角膜散光的变化

目的评价角膜地形图引导的透明角膜小切口超声乳化白内障吸除折叠式人工晶体植入术后角膜散光的变化。方法将119例(130只眼)白内障患者分为A、B、C三组,A组为对照组,B组为循规性散光组,C组为逆规性散光组;A、B两组行上方透明角膜切口,C组行颞侧透明角膜切口,行超声乳化白内障吸除折叠式人工晶体植入,比较术后角膜散光的变化情况。结果A、B、C三组术后90d平均手术性角膜散光度分别为(0.64±0.65)D、(0.75±0.58)D和(0.69±0.55)D,两两比较无显著性差异(P>0.05);平均角膜散光度A组较术前增加0.07D,B、C两组较术前减少0.34、0.37D,与A组比较有显著性差异(P<0.05)。结论以透明小切口行超声乳化白内障吸出折叠式人工晶体植入术,术后角膜散光度小,角膜地形图可准确反映角膜曲率变化,对指导术前角膜切口位置的选择及评价白内障术后角膜散光的变化具有重要的临床意义。     

Ocular injuries following sulfur mustard exposure—Pathological mechanism and potential therapy

Sulfur mustard (SM) is a potent vesicant, known for its ability to cause incapacitation and prolonged injuries to the eyes, skin and respiratory system. The toxic ocular events following sulfur mustard exposure are characterized by several stages: photophobia starting a few hours after exposure, an acute injury phase characterized by inflammation of the anterior segment and corneal erosions and a delayed phase appearing following a clinically silent period (years in human). The late injury appeared in part of the exposed eyes, expressed by epithelial defects and corneal neovascularization (NV), that lead to vision deficits and even blindness. During the last years we have characterized the temporal development of ocular lesions following SM vapor exposure in rabbits and have shown the existence of two sub-populations of corneas, those exhibiting delayed ocular lesions (clinically impaired) and those exhibiting only minor injuries if at all (clinically non-impaired). The aim of the present study was to investigate the pathological mechanism underlying the delayed injury by focusing on the unique characteristics of each sub-population and to test the efficacy of potential treatments.     

Effects of tumor necrosis factor-alpha, epidermal growth factor and transforming growth factor-alpha on interleukin-8 production by, and human rhinovirus replication in, bronchial epithelial cells.

Previous studies have shown that the epidermal growth factor (EGF) and transforming growth factor-alpha (TGF-alpha), members of the EGF growth factor family, regulate processes involved in airway repair after injury. Our studies were conducted to determine whether EGF and TGF-alpha directly control processes involved in the pathogenesis of airway inflammation and/or modulate the inflammatory effects of TNF-alpha in the airway mucosa. We have found that these cell growth factors directly induced interleukin-8 (IL-8) production by an immortalized human bronchial epithelial cell line (BEAS-2B cells). Furthermore, EGF and TGF-alpha enhanced tumor necrosis factor-alpha (TNF-alpha) induced IL-8 production by BEAS-2B cells. On the other hand, EGF but not TGF-alpha increased viral replication by BEAS-2B cells exposed to HRV-14. Moreover, TNF-alpha increased viral replication in the presence of EGF but not TGF-alpha. In conclusion, this study demonstrates that both EGF and TGF-alpha play an active role in the pathogenesis of airway inflammation.     

依诺沙星滴眼液治疗兔眼细菌性角膜溃疡的疗效观察

选用家兔角膜实层内注射菌液,制备膜溃疡病理模型。考察依诺沙星滴眼液对兔眼角膜溃疡的研究,以市售氟沙星滴眼液作阳性对照,生理盐水作阴性对照。结果表明,０．３％以上浓度依诺沙星滴眼液对Ｇ＾＋和Ｇ＾－菌所致角膜溃疡有较好的治疗作用。     

益生性肠球菌安全性评价研究进展与监管

肠球菌目前作为益生菌的研究热点,被应用于多种微生态制剂,在胃肠道疾病等多种疾病的治疗和预防过程中发挥着重要作用。但肠球菌在某些条件下可能成为致病菌,在临床报告的数据中,有一部分院内感染就是由肠球菌引起。鉴于肠球菌的益生性和潜在的致病风险,迫切需要对肠球菌的安全性进行评价。目前国内外的研究主要从耐药性和毒力因子方面评价肠球菌的安全性,也有部分研究通过动物试验考察肠球菌的安全性,结果均表明,尚未发现益生性肠球菌的安全问题。     

米诺环素对动脉瘤性蛛网膜下腔出血后神经保护作用的研究进展

米诺环素是四环素类广谱抗菌药,临床上用于治疗常见敏感病原菌引起的感染。近期研究发现米诺环素对动脉瘤性蛛网膜下腔出血后损伤体现出神经保护作用。米诺环素主要通过抗炎、抗凋亡、抑制基质金属蛋白酶-9等抗菌外作用减轻动脉瘤性蛛网膜下腔出血引发的早期脑损伤和迟发性脑血,且由于其良好的安全性和耐受性,对动脉瘤性蛛网膜下腔出血的辅助治疗具有一定潜力。本文就米诺环素对动脉瘤性蛛网膜下腔出血的神经保护作用及其可能机制进行综述,为后续深入研究及设计开展临床试验奠定基础。