角膜移植术后角膜在共焦显微镜下的形态学改变

目的：研究角膜移植术后角膜组织在共焦显微镜(Confocal microscopy)下的形态学改变。方法：应用Conoscan2．0共焦显微镜对板层角膜移植术后3～7d患者12例(12只眼)，术后1a患者8例(8只眼)，穿透性角膜移植术后3～7d患者10例(10只眼)，术后1a患者11例(11只眼)进行扫描检查，记录各层角膜图像。结果：板层角膜移植术后3～7d，植片中基质细胞较小，可见裂隙状暗纹和细小神经，层间为大面积高反光区，有点状颗粒沉积，植床水肿，细胞成像不清。移植术后1a，植片中未见神经，层间反光明显减弱，但仍有点状高反光颗粒沉积，植床中出现粗大裂隙状暗纹，内皮细胞密度正常。穿透角膜移植术后3～7d，植片中基质细胞“激活”，可见神经和后基质层的粗大暗纹，内皮细胞密度正常，细胞间可镶嵌有高反光点。术后1a，植片中基质细胞仍较小，未见神经，后基质层仍有裂隙状暗纹，内皮细胞体积增大，密度减少，高反光点消失。结论：Confoscan 2．0共焦显微镜可活体检查角膜移植术后角膜组织结构和细胞的病理改变，这对评估手术效果和临床观察以及跟踪随访具有重大意义。     

甘油保存角膜材料行深板层角膜移植术后临床和共焦显微镜观察

目的 观察甘油保存角膜材料行深板层角膜移植术后临床和共焦显微镜活体组织学改变.方法 2008年1月至2011年5月在我院行深板层角膜移植治疗的患者26例（26只眼）,所有患者植片均选择甘油脱水低温保存角膜材料,术后裂隙灯观察角膜植片愈合和透明情况,Cochet-Bonnet角膜知觉计测量角膜中央知觉值,共焦显微镜观察角膜全层组织的活体改变.结果 （1）裂隙灯观察26例患者均于术后1~3个月上皮完全修复,角膜植片透明,术后2例发生基质排异反应.（2）术后3个月可不同程度检测到角膜中央知觉,术后3个月角膜中央知觉平均阈值（18.35±9.56）mm,12个月（51.62±6.48）mm.（3）共焦显微镜观察术后1~3个月上皮细胞密度达正常值,3个月内未观察到角膜基质细胞,6~12个月基质细胞密度增加,术后6个月可观察到基质内神经干细胞,角膜内皮细胞密度维持稳定,但在两例发生基质排异反应患者出现内皮细胞密度明显下降.结论共焦显微镜在组织学上证明甘油保存角膜材料行深板层角膜移植后可在一定程度上恢复正常生理结构,并与临床观察角膜愈合过程相吻合.     

角膜内皮炎共焦显微镜形态学分析

目的:观察角膜内皮炎的共焦显微镜下形态学特征。方法:应用Confoscan 4.0共焦显微镜对24例24眼角膜内皮炎患者的角膜进行扫描检查,记录并分析各层角膜图像。结果:所有患者前部基质混浊,角膜深基质层可见基质细胞排列紊乱及条索状高反光结构,深基质层中还可见低反光带为后弹力层皱褶,角膜内皮细胞前可见斑片状大小不等的高反光结构,病变区角膜内皮细胞水肿、变性呈无结构暗区,内皮细胞呈多形性改变。4眼角膜上皮细胞边界不清,排列疏松,细胞较大,细胞核呈高反光结构,其中可见泡状暗区。结论:共焦显微镜可活体检查角膜内皮炎患者角膜组织各层结构,起到类似病理组织切片的作用;角膜内皮炎以深基质层及内皮细胞层损害为特征;共焦显微镜检查对角膜内皮炎具有一定的参考价值。     

激光扫描共焦显微镜观察穿透角膜移植术后角膜的形态学特征

目的研究活体穿透角膜移植术后各层角膜组织的激光扫描共焦显微镜(Laser Scan-ning Confocal Microscope,LSCM)的形态学特征。方法应用海德堡视网膜激光断层扫描仪Ⅱ/Rostock角膜模块(HRTⅡ)对20例20眼穿透角膜移植术后40天～2年的患者进行检查,记录并分析其各层角膜图像。结果穿透角膜移植术后早期角膜上皮细胞排列欠规则;Bowman膜和Descemet膜在激光共焦显微镜下无细胞结构;所有患者浅基质层细胞均较深基质层密集;稳定角膜植片其内皮细胞排列紧密,形态为均一六边形结构;所有患者均未发现上皮下神经丛及基质神经;术后植片排斥者有其特殊形态学表现。结论激光扫描共焦显微镜可活体检查穿透角膜移植术后角膜各层组织结构和细胞的病理改变,为临床诊断及用药提供有价值的客观依据,并在跟踪随访方面具有重大意义。     

微型角膜刀联合纤维蛋白黏合剂行无缝线光学板层角膜移植术的实验研究

目的 探讨微型角膜刀联合纤维蛋白黏合剂行无缝线光学板层角膜移植术的可行性和安全性。方法 24只健康纯种新西兰白兔随机均分为两组,用微型角膜刀制作植片和植床,Ⅰ组应用市售纤维蛋白生物黏合剂Tisseel,Ⅱ组不用纤维蛋白生物黏合剂。术后3d、2周、1个月、3个月裂隙灯显微镜观察角膜透明度和排斥反应情况,自动验光角膜曲率仪检测角膜屈光状态的改变,共焦显微镜活体观察术后角膜的创伤愈合反应。同期取角膜标本进行常规苏木素-伊红(HE)观察,并用荧光显微镜观察层间新生胶原纤维的情况。结果 Tisseel组11只眼(92％)的植片在位,未用纤维蛋白生物黏合剂组所有植片均脱落。Tisseel组术后1个月角膜完全透明;共焦显微镜发现术后植片植床层间附近的基质细胞数量减少,而后基质细胞、内皮细胞数量和形态均无变化。HE染色显示,术后3d层间出现无固定形态的嗜曙红染色带,1个月时消失,荧光显微镜观察DTAF组织染色未见术后明显新生胶原纤维。结论 微型角膜刀联合纤维蛋白黏合剂行无缝线光学板层角膜移植术简便、安全,术后角膜的创伤愈合反应轻微,纤维蛋白黏合剂不影响角膜的光学性能。     

颗粒状角膜营养不良活体共焦显微镜形态学研究

目的研究颗粒状角膜营养不良角膜各层组织的共焦显微镜形态改变。方法应用Confoscan2.0共焦显微镜对13例(26眼)颗粒状角膜营养不良患者的角膜进行扫描检查,记录与分析各层角膜图像。结果所有患眼前基质细胞及16/26眼后基质细胞结构不清,排列紊乱,并可见短棒状多形性强反光;6/26眼前弹力层不规则并增厚,神经纤维密度明显下降;6/26眼角膜上皮基底细胞层可见不定型的强反光;2/26眼角膜上皮细胞边界不清,排列呈疏松的蜂窝状,并出现不透明的强反光;所有患者角膜内皮细胞形态基本正常。视力0.3以下的患眼角膜上皮细胞层、上皮基底细胞层、前弹力层、后基质层发生形态异常的比例高于0.3以上的患眼(P<0.05)。结论1.共焦显微镜可活体检查颗粒状角膜营养不良角膜组织各层结构,起到类似病理组织切片的作用。2.前基质层形态异常可能是颗粒状角膜营养不良最基本的共焦显微镜形态特征,病情越重,前基质层以外的其它层次发生形态异常的可能性越大,但内皮细胞层一般不受累。3.共焦显微镜检查对颗粒状角膜营养不良手术方式的选择具有一定的参考价值。     

完全钝性基质层间分离的深板层角膜移植术

目的 介绍一种虹膜恢复器辅助的完全钝性基质层间分离的深板层角膜移植术,并评估应用该术式治疗各种角膜病的有效性和安全性.方法 通过激光共聚焦显微镜检查,选择角膜病变深度未达后弹力层,内皮功能正常的各类角膜病患者35例35只眼.通过虹膜恢复器辅助的完全钝性基质层间逐层分离的方式行深板层角膜移植,并对其疗效进行观察、分析.结果 35只眼中,2只眼(5.71%)有小的钝性撕裂,在保留少量深基质后均能继续完成手术;术后2只眼(5.71%)出现双前房,3d内均自行吸收;细菌和真菌性角膜溃疡术后均未复发.其中18只眼拆线后两周视力高于0.5(约占51.4%),25只眼高于0.3(约占71.4%).术后角膜地形图散光呈现递减的趋势,角膜内皮细胞密度无明显下降.术后6个月角膜植片均透明愈合,无明显层间混浊.结论 通过虹膜恢复器辅助的基质层间钝性分离的深板层角膜移植术是一种安全有效的手术方式.激光共焦显微镜检查可活体检查角膜各层组织结构,可为深板层角膜移植术提供良好的客观依据.     

激光共焦显微镜观察兔眼角膜缘干细胞缺乏羊膜移植后的形态

目的:评价激光共焦显微镜在观察兔眼角膜缘干细胞缺乏后创面修复过程中的作用.方法:角膜缘干细胞缺乏家兔18只(雌雄不限)分A组为正常眼组,B组为角膜缘干细胞缺陷羊膜移植组.两组的家兔于术后第1,3,5,7,10,15,30d进行中央角膜各层组织激光共焦显微镜检查.结果:通过连续共聚焦扫描,获取手术前后角膜各层平面图像(x,y轴)及纵向断层图像(z轴),图像均非常清晰.角膜表皮细胞排列疏松,呈多角型,胞体发暗,边界清晰发亮,明亮的细胞核可见或不可见.术后30d后角膜上皮细胞排列整齐,光反射正常;前基质细胞边缘欠清,胞核明亮呈纺锤形.后基质角膜细胞核较长,相对前基质密度较低.术后30d基质细胞排列紊乱,散在的、反光较强的圆形物质,并产生反射较高的细胞外基质.一些细胞呈梭形,基质产生点状与线形交织的瘢痕组织;基质层内有少量的新生血管,见红细胞在血管内流动;内皮细胞呈规则的六边形,细胞边界呈黑色胞体发亮,看不到细胞核,边缘发暗.术后角膜内皮细胞形态及数量无明显变化.结论:激光共焦显微镜可对角膜进行无创伤、实时活体检查,具有高清晰度、确切的深度定位、时间动态观察、纵向断层扫描等优势,更可提供理想的角膜精确、可重复、深径度图像.     

人体角膜移植内皮排斥反应的共焦显微镜研究

目的：应用共焦显微镜探讨活体角膜内皮排斥反应的早期形态学特征和变化。方法：19例(19只眼)角膜移植术后发生内皮排斥反应的人群根据角膜内皮排斥反应轻重的不同分为轻、中、重三组。应用共焦显微镜(Confoscan2．0)对角膜植片进行检查，并分析其相关因素。结果：活体角膜内皮排斥反应特有的共焦显微镜改变主要包括：①早期发现在KP及其附近的内皮细胞面有少量的免疫细胞附着，紧靠KP处的角膜基质细胞出现轻度水肿，细胞排列较混乱，胞浆中出现少量空泡状暗区。②中期发现在内皮细胞表面可见较多的免疫细胞聚集并被完全激活，基质细胞明显水肿，细胞核不清楚，基质中有免疫细胞进入；可见由高反光的免疫细胞和不明成份的组织碎片混合组成内皮排斥线。③在晚期排斥的患者中可见大量“激活”的免疫细胞在角膜内皮细胞面聚集，内皮细胞几乎完全破坏；基质细胞明显水肿，部分被“激活”且伴有大量免疫细胞聚集。结论：①内皮排斥反应的早期能够发现角膜内皮面有免疫细胞沉积；②角膜内皮面的免疫细胞沉积与角膜内皮排斥反应的轻重成正相关；③角膜内皮排斥反应实际上是一个混合排斥反应的过程；④共焦显微镜对角膜内皮排斥反应具有实时、无创、敏感、准确以及可莺蒽、可比较、可活体观察等临床价值。     

准分子激光原位角膜磨镶术后弥漫性板层角膜炎的共焦显微镜观察

目的探讨准分子激光原位角膜磨镶术（LASIK）后弥漫性板层角膜炎（DLK）的临床病理学特征及发病机制。方法LASIK术后DLK患者30例（39只眼）（Ⅰ～Ⅳ期）,术后1、3、5、7d及1个月进行裂隙灯显微镜检查,术后3、7d及1个月进行共焦显微镜检查。结果Ⅰ期及Ⅱ期DLK的典型表现出现在术后3d,共焦显微镜观察所见：角膜板层切口前基质及层间可见大量直径12～20μm的圆形或卵圆形细胞,反光较强,散在分布或排列成行,细胞内可见偏心的高反光的核和低反光的细胞内结构。层间还可见大量直径8～12μm的圆形细胞,强反光,多聚集成簇或排列成行,细胞核形态不规则。术后7d上述细胞几乎消失。Ⅲ期DLK出现于术后3～5d,表现为前基质中的细胞浸润更浓密,层间无定形的高反光物质较明显。Ⅳ期DLK在术后5～7d出现明显的前基质结构模糊,高反光,角膜瓣全层皱褶,晚期形成大量高反光的瘢痕组织。结论LASIK术后弥漫性板层角膜炎是角膜瓣层间的炎性反应,主要病理学特征为角膜瓣层间的炎性细胞浸润,其发病是多种因素通过内源性途径和外源性途径共同作用的结果。     

Follow-up studies in pattern VECP in demyelinating diseases in children

Spectral sensitivity functions and the transient decrease of sensitivity to short wavelengths after the offset of yellow light (transient tritanopia) were measured by increment threshold techniques in patients suffering from hereditary macular degenerations. Color vision defects were determined by arrangement tests and the anomaloscope. Central areolar choroidal dystrophy was found to produce a mild protan defect and to reduce foveal spectral sensitivity throughout the visible spectrum by a factor of 100; it also abolishes transient tritanopia. Electroretinogram (ERG) was normal, electrooculogram (EOG) subnormal. Stargardt's disease, despite numerous fluorescent macular spots, does not abolish transient tritanopia nor does it reduce spectral sensitivity, although scotopic matches were performed on the Nagel anomaloscope. Only in severe, advanced cases was transient tritanopia reduced and spectral sensitivity found to follow the absorption spectrum of rods. Routine ERGs and EOGs were normal. Vitelliform macular degeneration, despite the ophthalmoscopically pronounced dystrophic macula, produced only very small changes in spectral sensitivity and transient tritanopia, although a widened matching range on the Nagel anomaloscope and electrophysiological abnormalities were found. Apparently damage of the retinal circuit which connects long and short wavelength-sensitive cones, caused by hereditary conditions, is different from that caused by retinotoxic drugs.     

p53 expression in pterygium in two climatic regions in Turkey

Purpose:

To assess accumulation of p53 protein in samples of primary pterygium from people living in two different climatic regions in Turkey.

Materials and Methods:

Group 1 included 101 pterygium specimens from people in Adana located in southern Turkey. Group 2 included 39 pterygium specimens from people in Ankara, located in the middle of Turkey. Climatic conditions throughout the year are sunnier and warmer in Adana than they are in Ankara. The control group (Group 3) included 30 specimens of conjunctiva that had been excised during cataract surgery from 30 patients without pterygium. The pterygial specimens and control conjunctiva were studied by immunohistochemistry using antibodies against p53 protein. Pearson''s chi-square test was used to compare the p53 immunoreactivity.

Results:

The p53 immunoreactivity in Groups 1 and 2 was greater than it was in the control group (P<0.001). There were no differences in p53 immunoreactivity between Groups 1 and 2 (P= 0.060).

Conclusion:

The p53 immunoreactivity was not correlated with ultraviolet irradiation exposure. The p53 immunoreactivity in our pterygium specimens suggests that pterygium could be a result of uncontrolled cell proliferation.     

Changes in Bruch's membrane in experimental hypercholesteremia in rats

PURPOSE: We investigated the effect of high cholesterol diet for the aging changes in Bruch's membrane of rats. METHODS: After feeding a 4% cholesterol diet for 15 weeks to three young rats 3 months old and four aged rats 23 months old, we observed the morphological changes of Bruch's membrane by electron microscopy, and made a comparison with rats fed an ordinary diet. RESULTS: In one young rat fed a high-cholesterol diet, the endothelial basement membrane of the choriocapillaris formed multiple folds separated from the plasma membrane of the endothelium and showed lamellar thickening and crack in some areas. The elastic fiber layer in Bruch's membrane disappeared partly and some new microfibrils appeared. In one aged rat fed a high-cholesterol diet, the endothelial basement membrane of the choriocapillaris showed more lamellar thickening with lumps in some parts. Compared with rats fed an ordinary diet, rats fed a high-cholesterol diet showed thickening of the basement membrane and the changes were more severe. CONCLUSIONS: Our data indicated that high-cholesterol diet might promote age-related changes of Bruch's membrane.     

Trends in Patterns of Anterior Uveitis in a Tertiary Institution in Singapore

Purpose: To report the trends in etiology of patients with anterior uveitis (AU) in Singapore over 6 years.

Methods: A retrospective review of the clinical records of all new patients who presented with anterior uveitis to the uveitis subspecialty clinic from 2005 to 2010 at Tan Tock Seng Hospital, Singapore.

Results: There were 552 new cases of AU. This comprised 59.5% of a total of 928 new patients diagnosed with uveitis from 2005 to 2010. The mean age was 48.0?±?17.2 years. There was a male predominance (62.5%), with a male:female ratio of 1.7:1. The majority were of Chinese ethnicity (69%), followed by Malays (13.2%). Most cases were unilateral (79.5%) and idiopathic (50.4%). Common etiological causes included Fuchs heterochromic iridocyclitis (FHI) (5.6%), ankylosing spondylitis (AS)-related AU (5.1%), herpes simplex virus (HSV) (4.7%), and herpes zoster virus (HZV) (4.5%). There were increasing trends in AS-related AU from 3.2% in 2008 to 6.5% in 2010, and psoriasis-associated AU from 1.7% in 2005 to 4.0% in 2008. There were decreasing trends in the incidence of FHI from 10.6% in 2006 to 4.7% in 2009. No change in incidence of viral etiologies was noted, but cytomegalovirus-related immune-recovery uveitis (IRU) comprised 7.4%. IRU showed an increasing trend from 1.7% in 2005 to 11.9% in 2007, then decreased to 3.3% in 2010. Using the Pearson chi-square test, there was no statistically significant association between ethnicities (Chinese, Malay, Indian) comparing infectious and noninfectious cases (p?=?0.788), idiopathic and nonidiopathic cases (p?=?0.170), or between the various etiologies of uveitis (p?=?0.168).

Conclusions: AU was the predominant form of uveitis seen at our centers. Infectious etiologies (18.5%) are the most common among nonidiopathic cases, with herpes viruses (9.2%) being most prevalent. Despite increased use of polymerase chain reaction (PCR) in the detection of microbial and viral DNA, there was no overall increase in detection of infectious causes for uveitis. The changes in CMV-related immune recovery uveitis from 2005 to 2010 could reflect a change in HIV management in Singapore.     

Orthokeratology practice in children in a university clinic in Hong Kong*

Purposes: The aim of this study was to analyse clinical data of children undergoing orthokeratology (ortho‐k) and to investigate patients’/parents’ perspective on ortho‐k via telephone interviews. Methods: Clinical records of children undergoing ortho‐k from a university optometry clinic were reviewed and the effects of ortho‐k on refraction, vision and cornea were investigated. A telephone interview was conducted to solicit patients’/parents’ perspective of the treatment. Results: One hundred and eight files were reviewed. Median age of the children was nine years (range six to 15); mean (±SD) pre‐treatment refractive sphere was ‐3.56 ± 1.49 D and the median refractive cylinder was ‐0.50 D (range zero to ‐4.25 D). Significant refractive spherical reduction (58 per cent), improvement in unaided vision and corneal topographical changes were noted after only one night of wear. No significant change in astigmatism was found. Corneal staining was the most commonly observed complication with ortho‐k and more than 80 per cent of patients were advised to apply ocular lubricants to loosen the lens before lens removal. Ortho‐k was mainly undertaken for myopic control and about 90 per cent of the respondents reported good/very good unaided vision after ortho‐k and ranked the treatment as satisfactory or very good. Lens binding and ocular discharge were the most frequently reported problems during the treatment. Conclusion: Under close monitoring, overnight ortho‐k is effective and safe for reducing low to moderate myopia and the treatment is well accepted by the children.     

内毒素诱导的大鼠葡萄膜炎的巨噬细胞中Toll样受体4的表达

目的 探讨在内毒素诱导的Wistar大鼠葡萄膜炎中Toll样受体4(TLR4)阳性细胞与虹膜组织中巨噬细胞的动态变化和分布.方法 实验研究.Wistar大鼠50只,用随机数字法随机分为5组,每组10只,分别为正常对照(0 h)组、6 h组、12 h组、24 h组及48 h组.除0 h组外其余各组均足垫部注射霍乱弧菌内毒素200μg,注射后于裂隙灯显微镜下观察双眼前节炎症反应变化.按实验分组于0、6、12、24、48 h处死大鼠.取虹膜一睫状体及脉络膜组织.通过葡萄膜铺片免疫组织化学方法检测TLR4和巨噬细胞的标记CD163的表达.人工计数虹膜中TLR4~+与CD163~+的细胞并计算细胞密度,计算圆形和多形性的CD163~+细胞占所有CD163~+细胞的百分比.进一步采用免疫荧光双标记检测TLR4和CD163共表达的情况.通过单因素方差分析分别对大鼠虹膜内阳性细胞密度以及圆形、多形性CD163~+细胞的百分比进行统计学检验.结果 正常大鼠虹膜睫状体组织不表达TLR4.6 h组有2只大鼠虹膜内可见少量TLR4~+细胞,12～48 h组所有大鼠虹膜内TLR4~+细胞明显增多(F=167.2,P＜0.001),虹膜内TLR4~+细胞密度分别为(506.1±39.5)个/mm~2(12 h组)、(492.3±54.5)个/mm~2(24 h组)及(663.8±150.2)个/mm~2 (48 h组).在注射LPS后12～48 h期间TLR4~+细胞形态无明显变化.0～48 h组大鼠虹膜内均有CD163~+细胞,0 h组圆形和多形性CD163~+细胞百分比为13%,12～48 h组其百分比约为80%,且圆形细胞主要位于虹膜基质层.免疫荧光双标记可见TLR4和CD163的共表达,TLR4位于细胞膜,CD163位于细胞质.5组大鼠脉络膜内均未见TLR4表达.结论 内毒素诱导的大鼠葡萄膜炎中虹膜内TLR4表达增高,部分虹膜固有巨噬细胞表达TLR4.TLR4可能在葡萄膜炎的发生发展中起一定作用.     

弱视眼底光学相干断层扫描的研究进展

弱视是由于视觉发育关键期内各种异常的视觉经验导致单眼或双眼最佳矫正远视力低于正常同龄儿童,而眼部无明显器质性病变。目前普遍观点认为,弱视的发病机理主要源于视皮层。近年来,光学相干断层扫描（OCT）作为一种先进的活体成像技术,促进了对视网膜形态结构的大量研究,同时也被应用到弱视的研究领域。陆续有不同的研究人员利用OCT发现弱视患者眼底视网膜、脉络膜等眼部结构存在改变。笔者将对弱视眼底OCT的研究进展做一综述。     

实验性糖尿病视网膜微血管病变的病理研究

目的：观察糖尿病视网膜病变（diabetic retinopathy,DR）的组织学改变。方法：应用光镜、免疫组织化学、电镜及组织化学电镜等技术,研究在不同时间点Spregue-Dawley(SD)大鼠视网膜毛细血管基底膜中的Ⅳ型胶原蛋白及层黏蛋白和视网膜毛细胞血管基底膜的厚度,以及其负电荷位点数目的变化。结果：随着糖尿病病程的发展,视网膜毛细血管基底膜下不断增厚伴有Ⅳ型胶原蛋白及层黏蛋白的增加,同时负电荷位点数目减少。结论：视网膜毛细血管基底膜增厚,Ⅳ型胶原蛋白及层黏蛋白的增加,负电荷位点数目减少可能是导致DR渗出性病变的病理基础。