角膜的

The results of corneal topography, that of keratometer and cycloplegic retinoscopy in measuring astigmatism before refractive surgery were compared, and the measuring results of corneal topography and that of keratometer for 72 eyes after photorefractive keratectomy and 95 eyes after laser in situ keratomileusis were also compared.

采用计算机辅助的角膜地形图、角膜曲率计及散瞳验光三种测量散光的方法，对屈光不正患者360只眼进行检查，同时对准分子激光角膜切削术（photorefractive keratectomy，PRK）后72只眼和准分子激光原位角膜磨镶术（excimer laser in situ keratomileusis，LASIK）后95只眼的角膜散光用角膜地形图和角膜曲率计进行测量并比较。

Thirty-two pathological corneal samples were collected from the patients during corneal transplantation, including keratoconus (3 cases), endothelial keratitis (1 case), corneal endothelial decompensation after cataract operation (4 cases), Mooren's ulcer (3 cases), corneal implant opacities (5 cases), herpes simplex viral leucoma (1 case) and perforation (3 cases), trachoma corneal leucoma (1 case), bacterial (1 case) and fungal (1 case) corneal perforation, chemical trauma (5 cases), and burn (2 cases).

32例角膜植片标本来自我院角膜移植取下的病变组织，其中园锥角膜3例；角膜内皮炎1例；白内障术后角膜内皮失代偿4例；蚕食性角膜溃疡3例；角膜植片混浊5例；单疱病毒性白斑1例和单疱病毒性角膜穿孔3例；砂眼角膜白斑1例；细菌性和真菌性角膜穿孔各1例；角膜化学伤5例和热烧伤2例。

A corneal abrasion with a diameter of 7 mm was created centrally in both eyes with a trephine and subsequent rubbing off on the corneal epithelium with a 20% alcohol soaked swab. Photographic documentation and hematoxylin and eosin stained pathological section were performed when rabbits were killed at 0,12,24,36,48 and 72 h. Planimetry was performed, and the corneal photographs were analyzed with computer software.

将4组兔双眼角膜中央置入直径7.00mm的环钻，其内滴入20%乙醇约60s后用角膜刮匙刮去角膜上皮，制作角膜上皮缺损模型，术后每组滴用眼液频率均为3次／d，各组分别于术后12，24，36，48及72h行裂隙灯下荧光素染色照相及海德堡共焦激光角膜显微镜检查后各处死2只兔，取角膜做病理切片同时应用计算机图像处理系统测量角膜损伤后愈合面积。

Methods A lamellar flap was produced in the rabbit cornea with a microkeratome. The flap interface in whole corneas and corneal beds were soaked in the rAAV –EGFP solution for 10 min, BSS solution was applied in the control group. The corneas were harvested at day 1, 3, 14, and 21 after application for the structure analysis of cornea by HE staining. Expression of the transferred gene was monitored by fluorescence stereoscope and laser confocal microscope. Results After 3d of infection with rAAV-EGFP ,slight fluorescence was detected in rabbit cornea by fluorescence stereoscope .

微型角膜刀制作兔角膜基质瓣；转染组用含rAAV－EGFP转染液浸泡角膜瓣和角膜基质床10 min，对照组用等量BSS液浸泡角膜瓣和角膜基质床10 min，不同时间点(1、3、7、14、21d)通过荧光体视镜观察角膜出现EGFP荧光的起始时间及分布情况，收集角膜，其中一半用激光共聚焦显微镜观察、照相，另一半固定后行HE染色。

Repeated corneal perforation occurred in nine eyes, with primary diseases of herpes simplex keratitis (HSK; four eyes), Mooren ulcer, necrotizing keratitis and scleratitis, bacterial keratitis, and alkali burn. Corneal grafts perforated in 31 eyes, resulting from recurrent HSK, implant autoproteolysis, bacterial infections, recurrent Mooren ulcer, immunologic rejection, trauma, and fungal recurrence.

本研究中有9只眼发生了反复的角膜穿孔，其原发疾病分别为单疱病毒性角膜炎（HSK，4只眼），蚕食性角膜溃疡（2只眼），坏死性角巩膜炎（1只眼），细菌性角膜炎（1只眼）和碱烧伤（1只眼）。31只眼发生了角膜植片穿孔，分别由单疱病毒性角膜炎复发（8只眼），角膜植片自溶（7只眼），细菌感染（6只眼），蚕食性角膜溃疡复发（4只眼），免疫排斥反应（3只眼），外伤（2只眼）和真菌复发（1只眼）引起。

However, epikeratophakia, deep lamellar keratoplasty, intrastromal corneal ring, corneal crosslinking, and penetrating keratoplasty are the surgical way for the keratocous that contact lens could not correct.

角膜表层镜片术、深板层角膜移植术、角膜基质环植入术、角膜交联疗法及穿透性角膜移植术是临床上常用的手术治疗方法。

The experimental group corneas were preserved by organ culture for 4 weeks, the corneal thickness was measured with ultrasonic corneal pachymeter. Then every corneas were divided into half -chip, there are 48 half-chip total. It was divided into 4 groups, there are 12 half-chip in every groups. The corneal endothelial cell density of 12 half-chip were counted through Alizarin Red-Trypan blue staining; 12 half-clip corneas were fixed with 4% neutral formalin solution, HE staining was performed, the expression of AQP-1 in corneal stroma and corneal endothelial cell were detected through immunohistochemical staining; Na~+-K~+-ATPase activities in 12 half-clip corneas were examined with Na~+-K~+-ATPase kit; the expression of AQP-1 mRNA were detected through real-time fluorescent quantitation PCR.

实验组经器官培养保存4周后以角膜测厚仪测量角膜厚度，然后每个角膜被分成两半，共48个半片角膜，再分成4组，每组12个半片。12个半片用茜素红-台盼蓝染色染色行角膜内皮细胞计数；12个半片角膜用4%中性福尔马林溶液固定行HE染色、应用免疫组化染色检测AQP-1在角膜基质和内皮细胞表达的改变；12个半片角膜用Na~+-K~+-ATP酶试剂盒测量角膜内皮细胞Na~+-K~+-ATP酶活性；12个半片角膜用实时荧光定量PCR检测AQP-1mRNA表达改变。

The general situations of the eye were observed and the corneal thickness were measured with ultrasonic corneal pachymeter after the animal models was established. After a week, the corneas were removed after the experimental animals are put to death. The corneal endothelial cell density of 12 half-chip were counted through Alizarin Red-Trypan blue staining; 12 half-clip corneas were fixed with 4% neutral formalin solution , HE staining was performed, the expression of AQP-1 in corneal stroma and corneal endothelial cell were detected through immunohistochemical staining; Na~+-K~+-ATPase activities in 12 half-clip corneas were examined with Na~+-K~+-ATPase kit; the expression of AQP-1 mRNA were detected through real-time fluorescent quantitation PCR.

术后观察眼球大体情况、测量角膜厚度。1周后处死实验动物取角膜，用茜素红-台盼蓝染色染色行角膜内皮细胞计数；用4%中性福尔马林溶液固定行HE染色、应用免疫组化染色检测AQP-1在角膜基质和内皮细胞表达的改变；用Na~+-K~+-ATP酶试剂盒测量角膜内皮细胞Na~+-K~+-ATP酶活性；实时荧光定量PCR检测AQP-1mRNA在角膜内皮细胞表达的改变；并于正常对照组角膜比较。

MTT assay FAK signaling pathway inhibitor genistein on human corneal epithelial cell cytotoxicity;RT-PCR detection of human corneal epithelial cells adhesion to fungus at different times,extracellular matrix protein including laminin,fibronectin,FN glass,Ⅳcollagen,transmembrane protein integrinαⅤ,integrinβ1(ITGβ1),as well as the FAK signaling pathway FAK1,FAK2 and Paxillin gene expression;Western blot detection of the signal transduction pathway adhesion-associated protein ITGβ1,FAK and PAX expression and the inhibition of genistein. Immunocytochemical method was used to observe the LN,FN and FAK expression in human corneal epithelial cells during interaction with the fungues;Laser scanning confocal microscope had a cell positioning on FN,FAK and PAX,observed the changing of the human corneal epithelial cytoskeleton after stimulated by fungues;Quantitatived by flow cytometry to detect of human corneal epithelial cells with PAX at ITGβ1 fungal expression after adhesion;Optical microscopy quantitied the fungues and human corneal epithelial cell adhesion and recorded to determination the integral optical density afrer adhesion;Scanning and transmitted electron microscope observed the changing of cell ultrastructure after fungues and human corneal epithelial cell adhesion.

第一部分真菌激活人角膜上皮细胞FAK信号转导通路的体外实验研究将三种常见致病真菌(白色念珠菌、烟曲霉菌和茄病镰刀菌)分别与人角膜上皮细胞共孵育，MTT法检测FAK信号通路抑制剂染料木黄酮的对人角膜上皮细胞的细胞毒性作用；RT-PCR检测真菌黏附人角膜上皮细胞后不同时间细胞外基质层连蛋白、纤连蛋白、玻连蛋白、Ⅳ型胶原、跨膜蛋白整合素αV、整合素β1(ITGβ1)，以及FAK信号通路中FAK1、FAK2和桩蛋白基因的表达情况；Western blot的方法检测黏附信号转导途径相关蛋白ITGβ1、FAK和PAX的表达，以及染料木黄酮对真菌刺激人角膜上皮细胞FAK信息通路活化的抑制作用；免疫细胞化学方法观察人角膜上皮细胞与真菌相互作用过程中LN、FN和FAK的表达；激光共聚焦显微镜对FN、FAK和PAX进行了细胞定位，并观察真菌刺激后人角膜上皮细胞骨架的变化；流式细胞仪定量检测人角膜上皮细胞ITGβ1与PAX在真菌黏附后表达的改变；光学显微镜观察真菌与人角膜上皮细胞黏附数量，记录并测定了黏附后积分光密度值OD扫描及投射电镜观察了真菌与人角膜上皮细胞黏附后，细胞超微结构的改变。

The results are unsure for experimental use;The rabbit's corneas that were removed with upper-half of corneal limbal epithelium lamella and erased the center corneal epitheliums were transparent with intact corneal epithelium;In the approach,the corneal and limbal epitheliums were burned with a cotton swab socked in 1 mol/L NaOH,there were 4 rabbits' corneal stroma happened perforation or ulcer and symblepharon,and the other one presented corneal epithelium phenotype.This is an applicable method to create the pathological model of corneal limbal stem cell total deficiency.

结果表明，处理后4周，全周角膜缘上皮板层手术切除，中央角膜上皮层用1 mol/L NaOH擦除的5只试验家兔角膜表面全部血管化、结膜化，未发生睑球粘连，角膜基质胶原纤维完整未见溃疡、穿孔等病变，细胞印迹学检查为结膜表型，可作为实验性角膜缘干细胞移植的病理模型；全周角膜缘上皮板层手术切除，中央角膜上皮用生理盐水擦除的5只试验家兔，有2只为结膜表型，另3只为角膜表型，观察期内结果不稳定；半周角膜缘上皮板层手术切除，中央角膜上皮层用生理盐水擦除的5只试验家兔，角膜表面透明，全部为角膜表型；直接用1 mol/L NaOH擦除角膜缘和中央角膜上皮的试验家兔，有4只角膜基质胶原纤维断裂、溶解，并伴有严重的溃疡、穿孔、睑球粘连等病变，不能用于移植试验，另1只角膜表面透明，未见结膜和新生血管长入，细胞印迹学检查为角膜表型。

The split between the two groups can hardly be papered over.

这两个团体间的分歧难以掩饰。

This approach not only encourages a greater number of responses, but minimizes the likelihood of stale groupthink.

这种做法不仅鼓励了更多的反应，而且减少跟风的可能性。