仿LASEK法建立烟曲霉菌角膜炎动物模型的研究

目的 探讨建立真菌性角膜炎动物模型的新方法.方法 38只新西兰大耳白兔随机分为3组,右眼为实验眼.A组15只兔,采用准分子激光角膜上皮磨镶术(LASEK)制作直径7.0 mm、厚度50μm的角膜上皮瓣,瓣下接种25μL标准烟曲霉菌株(105孢子);B组15只兔,接种前7 d 0.1%地塞米松滴眼液点眼,每日4次,其余步骤同A组;对照组8只兔,以25μL生理盐水代替真菌菌液进行接种.术后采用裂隙灯显微镜、病理组织学检查、角膜真菌培养等方法观察评价角膜真菌感染情况.结果 A、B两组均见典型的真菌性角膜溃疡发生,经真菌培养鉴定为烟曲霉菌.对照组未见有角膜感染.A、B两组接种成功率分别为93.33%和100%,差异无统计学意义(χ2=1.345,P>0.05);B组角膜炎症较A组重、病程长,两组间临床评分差异有统计学意义(接种后3、7、15 d P<0.05,接种后30 d P<0.01).结论 采用LASEK制作角膜上皮瓣能够较理想地建立真菌性角膜炎动物模型.     

氟康唑加冰敷治疗真菌性角膜炎

目的探讨氟康唑加冰敷治疗真菌性角膜炎的效果。方法植物性眼外伤后真菌性角膜炎27例（27眼）分为两组：对照组14例,以氯康唑静脉滴注、结膜下注射及滴眼等方法治疗;冰敷组13例,除上述方法外再加冰敷治疗,观察治疗效果。结果对照组8例有效（57．14％）;冰敷组10例有效（76．92％）。冰敷组效果好（P〈0．05）。结论真菌性角膜炎氟康唑加冰敷治疗效果好。     

改良角膜表面镜片术法建立兔曲霉菌性角膜炎动物模型

背景真菌性角膜感染动物模型是研究真菌性角膜炎发病机制的工具,目前的制作方法主要有划痕法、基质注射法和角膜表面镜片术法,但均有其不足之处。目的探讨一种简便、易操作的改良兔曲霉菌性角膜炎动物模型制作方法。方法成年新西兰白兔18只,采用烟曲霉菌孢子附贴滤纸片的改良角膜表面镜片术法制作真菌性角膜炎动物模型。将浸有10^8孢子／ml（10^8孢子／ml组,6只）或10^6孢子／ml（106孢子／ml组,6只）真菌混悬液的滤纸贴敷于去上皮的角膜基质并用角膜接触镜覆盖,将浸有生理盐水的滤纸贴敷于另6只兔眼角膜作为对照组。分别于造模后3、7、14d裂隙灯下观察眼前节症状,参照Dong的标准进行症状评分。制备角膜刮片并用质量分数10％KOH和荧光白染色在荧光显微镜下检测真菌菌丝,角膜组织切片分别行苏木精-伊红和过碘酸希夫染色,光学显微镜下观察角膜形态学改变和菌丝生长情况。对感染组织进行真菌培养以验证模型的质量。结果10^8孢子／ml组和10^6孢子／ml组真菌性角膜炎模型成功者分别为6眼和4眼,裂隙灯检查表明造模3d后10^8孢子／ml组眼前节症状明显重于10^6孢子／ml组,且随着时间的延长,炎性损伤逐渐转向增生期。造模后3d和7d,2组感染的兔眼症状评分明显高于对照组,差异均有统计学意义（P〈0．01）,10^8孢子／ml组兔眼的症状评分均明显高于10^6孢子／ml组,差异有统计学意义（P〈0．01）,造模后14d,10^8孢子／ml组兔眼的症状评分明显高于对照组,差异有统计学意义（P〈0．05）。造模后3d和7d,2组兔眼角膜刮片中均可见真菌菌丝。角膜组织病理学检查显示,造模3d和7d后10^8孢子／ml组可见炎性细胞浸润和角膜基质细胞坏死,并可见真菌菌丝,造模后14d可见新生血管长入。10^6孢子／ml组炎症轻于10^8孢子／ml组。真菌培养结果表明,造模后3d和7d时10^8孢子／ml组均见菌丝生长,而10^6孢子／ml组仅在造模3d时可见菌丝生长。结论改良角膜表面镜片术法可成功制备兔曲霉菌性角膜炎动物模型,是一种简便、易于操作的真菌性角膜炎动物模型制作方法。     

结膜下注射AMD3100对小鼠角膜碱烧伤血管新生的治疗作用

目的探讨结膜下注射AMD3100对小鼠角膜碱烧伤血管新生的治疗作用及其机制。方法采用碱烧伤法诱导小鼠角膜血管新生（CNV）形成。在碱烧伤后当天,治疗组和对照组分别在球结膜下注射AMD31005μg（10μL）和生理盐水（10μL）,每日1次,连续用药7d。裂隙灯显微镜下观察不同时间点角膜炎症指数。病理组织学检查观察炎症细胞数量,并采用免疫组化检测角膜组织微血管密度。结果实验组在碱烧伤后不同时间点的炎症指数、炎症细胞数量和微血管密度均明显少于对照组（P〈0．05）。结论AMD3100可有效抑制CNV,可能与减轻碱烧伤后炎症反应有关。     

糖皮质激素对真菌性角膜炎预后的影响

探讨在确诊为真菌性角膜炎之前使用糖皮质激素对真菌性角膜炎患者的视力预后及治疗方式的影响。方法：回顾性病例对照研究。收集2014年12月至2015年12月于山东省眼科医院确诊的真菌性角膜炎患者211例（211眼），其中36例（36眼）在确诊为真菌性角膜炎之前的治疗中有明确使用过糖皮质激素的患者作为激素组，175例（175眼）在治疗过程中从未使用过激素者作为对照组。采集患者的人口学信息、临床检查结果、抗真菌药物治疗后的病情变化、治疗方案和治愈后2个月的最佳矫正视力（BCVA）。采用独立样本t检验与卡方检验对数据进行统计分析。结果：2组患者人口学特征对比差异无统计学意义。激素组发病时间为（14.5±10.1）d，明显短于对照组[（20.6±22.5）d]，差异有统计学意义（t=2.657，P=0.008）。激素组病灶大小为（6.3±2.4）mm，明显大于对照组[（4.8±2.1）mm]，差异有统计学意义（t=3.683，P<0.001）。激素组角膜刮片阳性率为97.2%，对照组阳性率为90.6%，差异无统计学意义（χ2 =1.633，P=0.201）。激素组有1例（2.8%）为药物治愈，明显低于对照组[34例（19.4%）]，差异有统计学意义（χ2 =5.983，P=0.014）；激素组中行穿透性角膜移植术的患者22例（61.1%），明显高于对照组[55例（31.4%）]，差异有统计学意义（χ2 =11.351，P=0.001）。激素组中（BCVA）低于 0.3的患者32例（88.9%），明显高于对照组[110例（62.8%）]，差异有统计学意义（χ2 =9.194，P=0.002）。结论：真菌性角膜炎使用糖皮质激素后病损范围增大，抗真菌药物治愈率低，穿透性角膜移植手术率高，预后不佳。     

更昔洛韦凝胶治疗带状疱疹病毒性角膜炎的临床疗效

目的观察更昔洛韦眼用凝胶治疗带状疱疹病毒性角膜炎的临床疗效。方法将门诊带状疱疹病毒性角膜炎患者随机分为治疗组22例（22只眼），局部点用更昔洛韦眼用凝胶，4次／d，疗程7d；对照组19例（19只眼），局部点用阿昔洛韦滴眼液，每2h1次，疗程7d。用药前及用药后每天观察病情变化。结果治疗组与对照组有效率分别为90．91％和63．16％，两者比较差异有非常显著性（P〈0．05）；治疗组角膜损害修复时间及虹睫炎临床症状消失时间均短于对照组。结论更昔洛韦眼用凝胶是治疗眼部带状疱疹安全有效的药物。     

羊膜移植对实验性HSK中基质金属蛋白酶表达的影响

目的研究羊膜移植（AMT）对单纯疱疹性角膜炎（HSK）中基质金属蛋白酶（MMP-2，-9）表达的影响。方法40只BALB／c小鼠角膜感染Ⅰ型单纯疱疹病毒（HSV-1），实验组角膜行AMT。术后第0、2、7、14d取出角膜。常规病理切片、免疫组化染色和计算机图像分析检测角膜中MMP-2及-9的表达及平均光度值的变化。结果对照组20只鼠眼中17只发生HSK；AMT组仅有9只眼发生，差异有显著统计学意义（P〈0．01）。AMT组角膜上皮、基质病变程度及新生血管发生率明显低于对照组（P〈0．05）。免疫组化及图像分析显示对照组角膜细胞和浸润炎性细胞中表达的MMP-2及-9在第2d增加，14d时达高峰。AMT组各时间点MMP-2及-9表达低于对照组，差异有显著统计学意义（P〈0．05）。结论羊膜移植可能通过抑制角膜细胞及浸润的炎症细胞产生MMPs，从而抑制HSK的发生和发展。     

泪液中（1，3）-β-D-葡聚糖水平检测对真菌性角膜炎的诊断和监测价值

背景真菌性角膜炎的早期诊断对有效治疗极为重要，但目前尚缺乏能够准确、定量诊断和判断疗效的客观指标。研究证实血浆中（1，3）一6一D一葡聚糖对全身真菌感染性疾病诊断有较高的敏感性和特异性，但泪液中（1，3）-β-D一葡聚糖含量的检测能否用于角膜真菌感染患者的早期诊断和病情监测尚不清楚。目的观察真菌性角膜炎患者接受抗真菌药物治疗过程中泪液中（1，3）-β—D一葡聚糖质量浓度的变化，客观评价其在真菌性角膜炎诊断和病情监测中的临床价值。方法选取2010年7月至2012年5月在青岛大学医学院附属医院眼科诊治的角膜溃疡直径≤5mm的真菌性角膜炎患者60例60眼为患病组，同期健康无眼疾的成年志愿者30人30眼为正常对照组。患病组接受抗真菌药物治疗，平均治疗时间为29d。分别在治疗前及治疗后7、14、28d和停药后7d、14d收集患眼泪液50斗l，进行（1，3）一p—D一葡聚糖质量浓度检测，同时结合激光扫描共焦显微镜检查及患者临床表现，对（1，3）一p—D一葡聚糖的诊断价值进行评估。对泪液中（1，3）一p．D一葡聚糖质量浓度低于20ng／L、激光扫描共焦显微镜检查未发现菌丝的患者，巩固治疗1周后停药，随访2个月。结果治疗前，患病组患者泪液中（1，3）-β—D-葡聚糖质量浓度为（Log）（6．37-＋0．48）ng／L，明显高于正常对照组（Log）的（2．00~0．31）ng／L，差异有统计学意义（t=2．89，P〈0．01）。真菌性角膜炎患者病情在治疗7d后开始好转，表现为溃疡边界逐渐清晰，病灶面积缩小，激光扫描共焦显微镜下显示菌丝密度比治疗前降低等。患病组患者泪液中（1，3）-β-D-葡聚糖质量浓度随着治疗时间的延长逐渐下降，其变化呈时间依赖性。治疗后7、14、28d患者泪液中（1，3）-β-D-葡聚糖质量浓度（Log）分别为（5．19±0．42）、（4．16±0．33）、（2．99±0．42）ng／L，停药后7d、14d分别为（2．91±0．39）ng／L、（2．80±0．40）ng／L，均明显低于治疗前的（6．37＋0．48）ng／L，差异均有统计学意义（P〈0．01）。治疗后28d至随访结束，患者泪液中（1，3）-β-D-葡聚糖保持稳定的低质量浓度，随访期间所有患者均无复发。结论作为一种定量检测方法，泪液中（1，3）-β—D-葡聚糖水平的检测可用于真菌性角膜炎的早期诊断及病情变化监测。     

高糖饮食对小鼠真菌性角膜炎的影响

目的　探究高糖饮食对小鼠真菌性角膜炎的影响。方法　选取健康无眼疾的雄性C57BL/6J小鼠78只，随机分为高糖饮食组和模型对照组，每组36只，模型对照组给予正常饮用水，高糖饮食组给予含体积分数10%果糖溶液，每2 d测量两组小鼠体质量及血糖，10 d后建立真菌性角膜炎模型。造模后24 h、36 h、48 h、72 h、96 h、120 h、168 h裂隙灯显微镜下对角膜进行临床评分并拍照。处死小鼠后，取角膜组织进行HE染色和PAS染色；测定角膜内中性粒细胞和巨噬细胞浸润体积。利用酶联免疫吸附实验对小鼠角膜内的白细胞介素-1β含量进行测定。结果　造模后 0~14 d，两组小鼠体质量与血糖差异均无统计学意义（均为P>0.05）。造模后24 h、36 h、48 h、120 h、168 h，高糖饮食组小鼠角膜临床评分均明显高于模型对照组，差异均有统计学意义（均为P<0.05）。高糖饮食组小鼠角膜穿孔率79.5%，高于模型对照组的40.9%，差异有统计学意义（P=0.000）。造模后各时间点，高糖饮食组中性粒细胞浸润体积均高于模型对照组，差异均有统计学意义（均为P=0.000）。造模后72 h、96 h、120 h、168 h，高糖饮食组巨噬细胞浸润体积均高于模型对照组（均为P=0.000）。角膜组织病理学检查结果示，高糖饮食组炎症反应更重，角膜组织破坏更早且更为严重。造模后24 h、48 h高糖饮食组白细胞介素-1β含量均明显高于模型对照组（均为P<0.05）。结论　高糖饮食加重了真菌性角膜炎感染的严重程度，增强了中性粒细胞、巨噬细胞的趋化，促进了IL-1β的分泌。     

维生素D_3对实验性糖尿病小鼠眼部结构的保护作用

背景研究证实,维生素D,通过其生物活化形式1,25-（OH）,D,与维生素D受体结合,从而发挥抗炎、免疫调节、神经保护作用。此外,维生素D,可以增强胰岛素的敏感性及促进胰岛素的分泌,控制高血糖症,减轻角膜水肿。目的研究维生素D,对缓解实验性糖尿病小鼠眼部结构损伤的作用。方法健康SPF级C57BL／6小鼠22只（体质量20-25g）,应用随机数字表法随机选取8只小鼠为维生素D,组,另取11只小鼠为糖尿病对照组,并取3只小鼠为正常对照组。利用腹腔注射质量分数2％链脲佐菌素（STZ）法制备实验性糖尿病模型,造模后2周维生素D,组小鼠行腹腔注射维生素D,5mg／kg,每周注射1次,共5次。各组每周进行血糖监测、直接检眼镜观察。各组每周随机摘取3只眼球,常规病理切片后于光学显微镜下观察眼球各部分结构,利用病理分析软件测量中央角膜、视盘旁1个视盘直径（DD）处脉络膜厚度及视网膜神经上皮层的厚度。结果糖尿病对照组小鼠造模后第1周角膜开始出现水肿,角膜平均厚度为（339．14＋11．13）μm,以后水肿持续但逐渐减轻（F=382．446,P=0．000）。维生素D,干预后水肿情况明显缓解,角膜厚度明显变薄（P〈0．05）。两组糖尿病小鼠第5周角膜上皮均开始萎缩（P〈0．05）,糖尿病对照组至第7周萎缩情况更加严重,而维生素D,组小鼠至第7周萎缩已明显减轻（P=0．002）。糖尿病小鼠脉络膜自第1周开始萎缩（P=0．010）,并随时间延长萎缩加重（F=437．411,P=0．000）。维生素D,干预后,第4周脉络膜萎缩趋势明显减缓,且用药后从第3周开始维生素D,组脉络膜厚度均大于糖尿病对照组（P〈0．05）,糖尿病小鼠自第1周开始视网膜神经上皮层即可见明显的凹凸不平,轻度水肿（P=0．000）,随时间延长,糖尿病造成的视网膜神经上皮层萎缩加重（F=91．859,P=0．000）。维生素D,组视网膜神经上皮层萎缩情况无明显改变。结论维生素D,对实验性糖尿病小鼠角膜及脉络膜有保护作用,但其对已萎缩的视网膜并无明显的修复作用。     

Influence of dimethylfumarate on experimental HSV-1 necrotizing keratitis

Background This study was performed to investigate the influence of fumaric acid esters on the course of herpes stromal keratitis (HSK).Methods The corneas of BALB/c mice were inoculated with 105 plaque-forming units of herpes simplex virus 1 (HSV-1, KOS strain). Groups of mice were treated intraperitoneally with phosphate buffered saline (PBS) (control mice), or with dimethylfumarate (DMF) at 15 mg/kg of body weight dissolved in PBS daily for 28 days pre-infection and for 14 days post-infection. The course of HSV-1 keratitis was studied clinically. Corneal sections were examined for inflammatory cell infiltration. The numbers of CD3, GR-1, CD11b and F4/80-expressing cells infiltrating the corneas were analyzed by immunohistochemistry.Results On day 14 after HSV infection, 72% of the mice in the control group had severe HSK. The development of HSK was reduced by DMF treatment in the DMF group (22%) (P=0.004). The total number of inflammatory cells and infiltration of polymorphonuclear-neutrophils (PMNs) were reduced in the corneas of DMF-treated mice. Compared to the PBS-treated mice, numbers of CD3, CD11b, GR-1 and F4/80-positive cells were reduced in the DMF group of mice.Conclusions The course of experimental herpes stromal keratitis can be improved with systemic fumaric acid ester treatment. The improvement of keratitis correlates with a reduced corneal infiltration of T cells and mononuclear cells.The authors have no financial interest in any of the reagents used in this study     

Herpetic keratitis in inbred mice

The corneas of four inbred strains of mice (BALB/c, DBA/2, C3H and C57BL/6) were inoculated with the RE strain of herpes simplex virus, type 1. The corneas were examined at frequent intervals and graded on a scale of 0 (clear cornea) to +5 (severe necrotizing stromal keratitis). At 3 weeks postinfection, the mean corneal scores were: DBA/2, 4.0; BALB/c, 2.2; C3H, 0.7; and C57BL/6, 0.15. The differences between the scores are statistically significant (P less than 0.05), except for the C3H and C57BL/6 strains. The order of severity of corneal disease in these mice corresponds to the order of susceptibility to systemic infection found in these same inbred strains. Additional studies of herpetic keratitis in inbred mice should prove helpful in understanding the genetic and immunologic basis of herpetic stromal keratitis.     

明胶酶在兔真菌性角膜炎病理改变中的作用研究

目的探讨明胶酶包括基质金属蛋白酶（MMP）2与MMP-9在兔真菌性角膜炎病理改变中的作用。方法80只新西兰白兔随机分为4组,每组20只。其中3组为实验组,兔右眼分别注入100μl茄病镰刀菌、烟曲霉菌及白色念珠菌的悬液;另1组为对照组,兔右眼注入等量生理盐水。免疫组织化学方法观察MMP-2与MMP-9的来源,明胶酶谱法检测其活性。组织病理学方法观察炎性细胞的浸润、角膜细胞外基质（ECMs）的降解以及真菌菌丝在角膜内的生长方式与入侵深度。结果MMP-2主要由角膜基质细胞产生,真菌感染后5d检测出活性,8d活性升高。MMP-9主要来源于嗜中性粒细胞,接种后1d即检测到活性,3d活性升高,之后逐渐下降。茄病镰刀菌感染后3d,角膜内散在嗜中性粒细胞,浅层ECMs被降解,菌丝平行于角膜基质纤维生长。烟曲霉菌和白色念珠菌感染后3d,角膜内可见大量嗜中性粒细胞,周围ECMs降解明显,菌丝表现为垂直生长。接种后8d,茄病镰刀菌和白色念珠菌感染的角膜内炎性细胞和菌丝明显减少,而烟曲霉菌感染的角膜变化不明显。结论茄病镰刀菌、烟曲霉菌及白色念珠菌感染兔角膜后,产生的明胶酶活性明显不同;明胶酶对降解角膜ECMs发挥了重要作用;随着ECMs降解程度的不同,菌丝在角膜内的生长方式、入侵深度等病理改变出现差异。     

Therapeutic action of meliacine,a plant-derived antiviral,on HSV-induced ocular disease in mice

Ocular herpes simplex virus type-1 (HSV-1) infections remain an important cause of corneal disease which may result in a loss of vision. Meliacine (MA), an antiviral activity present in crude leaf extracts of Melia azedarach L. that inhibits HSV-1 multiplication in vitro, was studied in a murine herpetic stromal keratitis experimental model. Adult Balb/c mice were inoculated with HSV-1 at their corneas after abrasion. MA was administered topically three times a day for 3 consecutive days, beginning at 24 and 96 hr after infection. Infected animals treated or not with MA were monitored for the development of ocular disease by a binocular microscope for 16 days. MA significantly reduced the incidence and the severity of blepharitis, neovascularization and stromal keratitis with respect to untreated infected mice, regardless the schedule of treatment assayed. Histological examination of corneas from MA-treated animals revealed no tissue damage, whereas corneal samples from untreated infected mice showed inflammation, vascularization and necrosis. In uninfected mice treated with MA, we found no evidence of corneal damage and histopathological studies showed no changes in the corneas of these mice. Treatment with MA at 24 hours post-infection (h.p.i.) reduced viral multiplication in the eye by 1-1.5 orders of magnitude. Studies on latency revealed that MA sligthly affected the establishment of a latent infection. Thus, MA proved to exert an antiviral action on the development of herpetic stromal keratitis when supplied by post-treatment. Unexpectedly, treatment with MA after 96h.p.i prevented ocular disease, suggesting an in vivo immunomodulating activity of MA.     

Incidence and microbiological profile of mycotic keratitis in a tertiary care eye hospital: A retrospective analysis

Purpose

To determine the incidence and microbiological profile of mycotic keratitis seen at a tertiary care eye hospital.

Materials and methods

A retrospective review of microbiology records of patients presenting with suspected microbial keratitis seen between January 2006 and December 2009 was performed. Patients with positive fungal cultures were further analyzed for the type of fungus isolated and associated bacterial pathogens.

Results

Microbiology records of 2300 patients with suspected microbial keratitis were reviewed. A microbiological diagnosis of mycotic keratitis was established in 87 (3.8%) patients over a four year period based on positive fungal cultures. The yearly incidence of mycotic keratitis was 3.2% (2006), 4.9% (2007), 3.3% (2008) and 3.6% (2009). Filamentous fungi were isolated more often than yeasts. Aspergillus species followed by Fusarium species and Trichophyton species were the commonest filamentous fungi isolated while Candida albicans was the most frequently encountered yeast. Mixed infections due to fungal and bacterial pathogens were seen in 25/87 (28.7%) patients.

Conclusion

Cumulative incidence of mycotic keratitis was 3.8% over a four year period. Aspergillus species and Candida albicans were the most frequent pathogenic organisms causing mycotic keratitis in this part of the world. Mixed infections were seen in 28.7% of the patients. Knowledge of the “local” etiology within a region may be valuable in the management of mycotic keratitis in instituting an empirical therapy, especially when facilities for microscopy, cultures and antifungal susceptibility are not readily available. The baseline information presented will also be helpful in the planning of a corneal ulcer management strategy and for future studies on mycotic keratitis.     

Experimental model of Fusarium solani keratitis in rats

AIM: To establish a repeatable rat model of Fusarium solani keratitis (F. solani keratitis) that mimicked fungal keratitis in humans. METHODS: Wistar rats’ corneas were scratched on the superficial stroma after scraping the unilateral corneal epithelia. Then, the corneal surface was inoculated with different inoculum dose of F. solani spore suspension. Doses ranged from 106 to 109 colony-forming unit per milliliter (CFU/mL). The treated corneas were covered by contact lenses that were made of Parafilm M membrane. Negative controls were inoculated with sterile phosphate-buffered saline (PBS). For statistical analysis, corneas were evaluated daily on a 12-point scale to check the state of corneal inflammation. Furthermore, the pathological characteristics of this model were investigated. RESULTS: The rat model of F. solani keratitis was established by the combination methods of corneal trauma and parafilm-made contact lens and inoculation of fungus spore suspension. 106 and 107CFU/mL of F. solani induced mild corneal infection, while 108CFU/mL of F. solani was sufficient to induce moderate infection that was consistent with human keratomycosis. Dose of 109CFU/mL of F. solani was excessive and led to perforated corneas. CONCLUSION: The rat model of F. solani keratitis, established by the combinational methods of corneal trauma, parafilm-made contact lens and the appropriate dose of inoculum, that imitates the developing processes of F. solani keratitis in human beings and provides a repeatable method of creating a rat model.     

Amnionmembrantransplantation bessert experimentelle herpetische Keratitis

PURPOSE: Transplantation of human amniotic membrane (AMT) accelerates the healing of experimental ulcerative herpetic keratitis. Here the expression and activity of matrix metalloproteinase (MMP)-9 was studied. METHODS: BALB/c mice were corneally infected with HSV-1. Whereas the infected corneas of mice in group 1 were covered with AM, tarsorrhaphies were performed in others (group 2). After 2 days, the appearance of corneal ulcers and stromal inflammation was judged clinically, and the corneal PMN infiltration was studied histologically. The expression of MMP-9 in the corneas was localized by immunohistochemistry and analyzed by Western-blot technique. The MMP-9 activity in the corneas was determined by zymography. RESULTS: On day 14, the ulcerating corneas had a dense PMN infiltration, the ulcers and the majority of PMNs were highly positive for MMP-9, and the active forms of MMP-9 were detected. Gelatinolytic activity was found in these corneas by zymography. Compared with the mice of group 2, ulceration, stromal inflammation and neovascularization markedly improved clinically and histologically within 2 days in mice of group 1. This was associated with a reduced expression of MMP-9 in corneal tissue and in PMNs. The gelatinolytic activity of MMP-9 was reduced after AMT. CONCLUSIONS: These observations suggest that improvement of herpetic corneal ulcers and reduced corneal neovascularization after AMT may result from a reduced expression and activity of MMP-9.     

Matrix metalloproteinases (MMP-8, MMP-9) and the tissue inhibitors of metalloproteinases (TIMP-1, TIMP-2) in patients with fungal keratitis

PURPOSE: To study the infiltrating cells and quantify the levels of matrix metalloproteinases (MMP-8, MMP-9) and tissue inhibitor of metalloproteinases (TIMP-1, TIMP-2) in the cornea, tear, and serum of patients with fungal keratitis. METHODS: Experimental study. Infected corneal tissue from 4 patients with fungal keratitis (group 1) scheduled to undergo therapeutic keratoplasty accounted for the histopathologic and cytospin smear analysis. Ten patients with fungal keratitis from group 2 served for the quantification of MMPs and TIMPs. Five patients with keratoconus undergoing penetrating keratoplasty and 5 cadaver corneas were chosen as controls for group 2. Corneal buttons obtained during keratoplasty, 15 to 20 microL of tears collected using the capillary flow method, and 3 mL of blood was obtained from patients with fungal keratitis and patients with keratoconus. Corneal button sections from group 1 were stained with hematoxylin and eosin and Grocott methenamine silver nitrate for the histopathologic studies and Giemsa staining for the cytospin smear analysis. Enzyme-linked immunosorbent assay was used for the quantification of total MMP-8, MMP-9, TIMP-1, and TIMP-2 in the corneal homogenates, tear, and serum samples of group 2. RESULTS: Corneal sections from group 1 revealed dense fungal filaments and a large proportion (91.4% +/- 38%) of polymorphonuclear leukocytes (PMNs). Significant elevation in the levels of MMP-8 and MMP-9 (P < 0.05) in the fungal keratitis corneas was observed in group 2 compared with the cadaver and keratoconus corneas. The ratio of MMP/TIMP was also higher in the fungal keratitis corneas. CONCLUSIONS: Infiltrating PMNs in the cornea of patients with fungal keratitis contributed to the increased activities of MMP-8 and MMP-9, thereby enhancing tissue destruction and derangement.     

真菌性角膜炎的实验观察与治疗

在兔角膜板层注射镰刀菌，形成深层真菌性角膜炎，病灶向浅层发展形成角膜溃疡。另外，可见虹膜炎、前房积脓、新生血管和卫星病灶，未见角膜穿孔。这些真菌性角膜炎用萘替芬治疗，每天用药５次，共治疗１３天。结果表明萘替芬有一定治疗作用，但与对照组相比差异性不够显著（Ｐ〉０．０５）。