实验性糖尿病兔角膜神经组织化学和超微结构观察

用四氧嘧啶诱发糖尿病兔模型，以观察糖尿病兔角膜神经分布的形态学改变。试验兔分别于６、８、１０、１２周各处死一批，用组织化学技术和透射电镜观察兔角膜神经的分布、形态和结构改变。结果发现，糖尿病兔自造模后６周，神经网丛密度明显稀疏，神经分支变细，神经活性降低。超微结构显示，糖尿病兔角膜神经轴突肿胀，其内神经原纤维分布不均，线粒体呈现不同程度的变性。随着病程延长，上述变化日趋明显。推测糖尿病时角膜神经的     

Ⅱ型糖尿病患者角膜细胞形态学特征的共焦显微镜观察

目的探讨II型糖尿病患者的角膜在共焦显微镜下各层细胞形态学特征。方法应用Confoscan3.0共焦显微镜对120例(146只眼)II型糖尿病患者和36例(36只眼)同年龄对照组的中央角膜进行检查,依据双目间接眼底镜和眼底荧光血管造影检查的结果,将糖尿病患者分为三组:糖尿病无眼底改变(NDRP)组、非增生性糖尿病视网膜病变(NPDR)组、增生性糖尿病视网膜病变(PDR)组,记录角膜各层细胞的图像,并对其进行分析。结果共焦显微镜下可以观察到糖尿病引起的角膜神经营养性上皮病变;和对照组比较,2型糖尿病患者角膜上皮下神经丛神经分支密度减少,神经纤维的密度在PDR组明显减少,NPDR组、NDRP组的减少不明显。角膜基质中形态异常的神经纤维在糖尿病组中出现的机率也明显高于对照组;角膜前基质细胞密度在PDR组有明显减少,后基质细胞在NPDR组和PDR组均明显降低,NDRP组减少不明显;糖尿病组的后弹力膜层病变表现为特征性的皱折形成;糖尿病各组角膜内皮细胞密度较对照组无显著性差异,但糖尿病各组的细胞面积变异系数高于对照组,六角形细胞所占的百分比在糖尿病各组均较对照组减少,且随着糖尿病视网膜病变的进展,六角形细胞的百分比亦逐渐减少,差异有统计学意义。结论共焦显微镜检查是一种有效、无创的角膜检查方法,糖尿病对角膜每一层结构均有重要影响,角膜上皮层、角膜神经、基质细胞密度、内皮细胞密度和形态均存在改变。     

2型糖尿病患者角膜知觉减退的研究进展

角膜知觉减退是由于多种原因造成的神经传导功能障碍。尽管角膜知觉减退是否是糖尿病的眼部症状还存在争议,但是最近有研究表明,它与糖尿病病程、血糖控制情况和糖尿病神经病变都存在相关性。近几年,角膜知觉减退作为糖尿病周围神经病变的一个监测指标成为研究热点之一。其机制可能是由于神经细胞内糖代谢产物累积、神经纤维数量减少和长度缩短、免疫机制所导致。我们现就角膜知觉减退与糖尿病的国内外研究进展做一综述。     

2型糖尿病患者角膜基质细胞形态学特征的共焦显微镜观察

目的 探讨2型糖尿病患者在共焦显微镜下角膜基质细胞形态学特征。方法 应用Confoscan3．0共焦显微镜对120例（146眼）2型糖尿病患者和36例（36眼）同年龄对照组的中央角膜进行检查,依据双目间接眼底镜和荧光素眼底血管造影（FFA）检查的结果,将糖尿病患者分为3组：糖尿病无眼底改变（NDR）组、非增生型糖尿病视网膜病变（NPDR）组、增生型糖尿病视网膜病变（PDR）组,记录角膜基质细胞和基质神经的图像,并对其进行分析。结果 与对照组比较,糖尿病各组角膜的中基质细胞密度均较对照组降低,而前基质细胞的密度只在PDR组有明显的减少,后基质细胞在糖尿病视网膜病变组均明显降低。角膜基质中形态异常的神经纤维在糖尿病组中出现的概率也明显高于对照组,两组间有显著统计学意义（,=36．450,P=0．000）。结论 共焦显微镜是一种有效无创的角膜检查方法;2型糖尿病患者角膜基质细胞密度减少,基质细胞、基质神经均存在形态学的异常。     

共焦显微镜观察2型糖尿病患者角膜神经分布及形态学特征

目的探讨2型糖尿病患者共焦显微镜下的角膜神经分布及形态学特征。方法应用Confoscan3．0共焦显微镜观察59例（65只眼）2型糖尿病患者和26例（26只眼）同年龄对照组白内障手术患者的中央角膜神经分布和形态学特征,依据双目间接检眼镜和荧光素眼底血管造影检查结果,将糖尿病患者分为3组：糖尿病无眼底改变（NDR）组、非增生性糖尿病视网膜病变（NPDR）组、增生性糖尿病视网膜病变（PDR）组,记录并分析患者角膜上皮下神经丛和角膜基质神经图像。结果与对照组相比,2型糖尿病患者角膜上皮下神经丛神经分支密度、神经纤维长度均减少,差异有统计学意义（P〈0．01）,神经纤维的密度仅PDR组明显减少,NPDR组、NDR组减少不明显。角膜基质中形态异常的神经纤维在糖尿病患者中出现的几率也明显高于对照组,两组差异有统计学意义（P〈0．01）。结论共焦显微镜是一种有效、无创的角膜神经检查方法,2型糖尿病患者角膜上皮下神经丛、角膜基质神经均显示形态学异常。（中华膑群杂志,2006,42：896—900）     

糖尿病性角膜感觉减退及其发病机理

糖尿病性角膜感觉减退及其发病机理同济医科大学附属协和医院李金瑛综述麦才铿，胡燕华审校糖尿病是一种常见且治疗很棘手的内分泌疾病，其在眼部的表现以糖尿病性视网膜病变和白内障最引人法目，角膜感觉减退常被忽略。角膜感觉是人体最敏感的防御反射之一，影响它的因素...     

2型糖尿病患者角膜上皮基底膜沉积物的激光共焦显微镜观察

目的 观察2型糖尿病患者角膜上皮基底膜沉积物在激光共焦显微镜下的表现,探讨糖尿病患者出现角膜上皮病变的可能因素.方法 应用激光共焦显微镜对9例(15眼)2型糖尿病患者和年龄、性别匹配的健康对照眼的全角膜进行分区扫描检查,记录角膜上皮层、前弹力层及前基质层的图像,同时行眼前节光学相干断层扫描(OCT)检查,对所有检查资料进行分析.结果 9例(15眼)中,激光共焦显微镜显示8例(11眼)出现角膜上皮基底膜的异常沉积物,部分沉积物周边可见纤细的神经纤维,角膜上皮层下神经纤维异常,神经纤维密度明显低于正常对照眼.眼前节OCT检查发现有4眼的基底膜、前弹力层及前基质层部位有强反射信号.角膜上皮基底膜沉积物与角膜上皮下神经纤维密度间无明显相关性(r=-0.265,P=0.430).结论 2型糖尿病患者角膜上皮基底膜出现异常沉积物,可能是糖基化终末产物(AGEs)在角膜上皮基底膜的沉积,对糖尿病角膜上皮病变及角膜上皮下神经病变有一定的影响.     

Ⅱ型糖尿病患者角膜的共聚焦显微镜观察

目的共聚焦显微镜下观察Ⅱ型糖尿病患者角膜上皮下神经纤维以及角膜各层细胞密度和形态学改变。设计比较性病例系列。研究对象选取确诊为Ⅱ型糖尿病的患者20例(20眼)为病例组和就诊于眼科门诊的无糖尿病的老年性白内障患者20例(20眼)为对照组。方法病例组和对照组均进行角膜共聚焦显微镜检查,对所得的图片和结果进行分析。主要指标角膜上皮下神经纤维密度、角膜上皮基底细胞密度、角膜浅基质细胞密度、角膜中基质细胞密度、角膜深基质细胞密度、角膜内皮细胞密度、内皮细胞的变异系数和六角形百分比。结果糖尿病组与对照组相比,上皮下神经纤维密度明显减少(t=-4.951,P=0.000);角膜浅基质细胞密度、中基质细胞密度和深基质细胞密度均有显著下降(P值均=0.000);角膜内皮细胞变异系数增加(t’=3.652,P=0.001),六角形百分比减少(t=-3.778;P=0.001)。结论共聚焦显微镜下显示Ⅱ型糖尿病患者角膜上皮下神经纤维密度和各层细胞密度下降。     

角膜激光共焦显微镜在糖尿病视网膜病变中的应用

目的　探讨角膜激光共焦显微镜在观察糖尿病视网膜病变（diabetic retinopathy,DR）患者角膜上皮下神经丛、角膜细胞密度及形态变化中的价值。方法　选取确诊的DR患者94例（114眼）,其中非增生型糖尿病视网膜病变（non-proliferative diabetic retinopathy,NPDR）患者41例（52眼,NPDR组）、增生型糖尿病视网膜病变（proliferative diabetic retinopathy,PDR）患者53例（62眼,PDR组）,收治时间2016年1月至2017年4月,同期糖尿病无眼底改变的患者40例（40眼）作为对照组,三组均采用角膜激光共焦显微镜进行检测,对比各组角膜上皮下神经丛、角膜细胞密度及形态的变化。结果　NPDR组和PDR组患者的角膜基底层、浅基质层、中基质层、深基质层的细胞密度均显著低于对照组（均为P<0.05）;PDR组患者的角膜基底层、浅基质层、中基质层、深基质层的细胞密度均显著低于NPDR组（均为P<0.05）;NPDR组和PDR组患者的角膜内皮细胞密度、六边形细胞比例、神经纤维密度、神经纤维长度均显著低于对照组（均为P<0.05）,NPDR组和PDR组患者的内皮细胞变异率、神经分支密度均显著高于对照组（均为P<0.05）;PDR组患者的角膜内皮细胞密度、六边形细胞比例、神经纤维密度、神经纤维长度分别为（1962.0±117.3）个·mm^-2、46.1%±5.5%、（15.4±3.3）根·mm^-2、（6.2±2.7）mm·mm^-2,均显著低于NPDR组的（2381.4±144.0）个·mm^-2、58.2%±7.0%、（20.6±3.8）根·mm^-2、（8.6±2.4）mm·mm^-2（均为P<0.05）,PDR组患者的内皮细胞变异率、神经分支密度均显著高于NPDR组（均为P<0.05）。结论　角膜激光共焦显微镜能有效观察DR患者角膜上皮下神经丛、角膜细胞密度及形态变化,为临床诊断、治疗提供指导。     

正常人角膜不同位置角膜知觉的测定分析

人类角膜神经分布密集，大多数神经纤维为感觉神经。这些感觉神经对外界的机械、化学和热刺激所作反应的敏感程度即为角膜知觉。以往采用棉丝检测角膜知觉，近年来发明的CohetBonnet角膜知觉计，可使角膜知觉得以量化测定并分析。为探讨正常人角膜不同位置的角膜知觉分布情况，我们应用CohetBonnet角膜知觉计，检测126位自愿者的角膜中央、鼻侧、颞侧、上方及下方的角膜知觉，现将结果报告如下。     

增殖性糖尿病视网膜病变患者活体角膜组织学变化的研究

目的探讨增殖性糖尿病视网膜病变（PDR）患者角膜知觉、激光共焦显微镜下角膜上皮、神经纤维及内皮细胞的组织形态学变化。方法采用激光共焦显微镜对2006年6月至11月于玻璃体视网膜科住院的30例增殖性糖尿病视网膜病变患者进行角膜上皮、神经纤维、内皮细胞形态观察,并行角膜知觉检查;与同期白内障中心住院的同年龄30例非糖尿病患者对照。结果PDR患者角膜知觉较对照组明显减退（P〈0.001）。共焦显微镜检查显示,PDR组角膜上皮细胞、内皮细胞密度均低于对照组（P〈0.05）：角膜神经丛的密度明显低于对照组（P〈0.001）;PDR患者角膜神经纤维纤细,走行迂曲,分支减少多呈钝角。结论激光共焦显微镜已成为在活体状态下观察角膜形态的新方法,增殖性糖尿病视网膜病变患者角膜知觉减退与角膜神经纤维数目减少有关。     

In vivo confocal microscopy in hydroxychloroquine-induced keratopathy

Background Vortex keratopathy, arising as a side effect of several medications, is characterized by golden-brown deposits in the cornea. Methods A 41-year-old woman treated for sarcoidosis with hydroxychloroquine therapy and suffering from vortex keratopathy was examined by in vivo confocal microscopy. Scans of both corneas were performed. Results By slit lamp examination, the left but not the right eye showed a golden-brown deposit throughout the cornea. In vivo confocal microscopy revealed the presence of highly reflective, dot-like intracellular inclusions concentrated in the basal epithelial layer. They were also detected within the anterior and posterior stroma, but not within the endothelium. In regions of the anterior stroma, devoid of inclusions, hyperreflective ramified keratocytes were observed, forming an extended interconnecting network. Conclusion In addition to the granular deposits, in vivo confocal microscopy revealed hyperreflective, possibly phagocytic keratocytes.     

In vivo confocal microscopy of pre-endothelial deposits

BACKGROUND: Deposits in various layers of the cornea might result from long-term medical therapy, photorefractive surgery, and longterm use of contact lenses or corneal dystrophies. METHODS: A 46-year-old woman was referred to our department with the suspected diagnosis of posterior polymorphous dystrophy. Slit-lamp biomicroscopy revealed bilateral small-sized deposits in the posterior part of the cornea. In vivo confocal microscopy was performed to evaluate these deposits in detail. RESULTS: In vivo confocal microscopy of the cornea identified hyperreflec-tive "dot-like" structures in the deep stromal layer and anterior to the endothelial cell layer. The morphology and number of keratocytes of the posterior stroma and of endothelial cells appeared normal. CONCLUSIONS: In vivo confocal microscopy is a very useful tool to analyze and visualize pre-endothelial deposits. Because there is no family history of corneal disease, the exact origin of the pre-endothelial deposits in our case remains unclear.     

中国人正常角膜的共焦显微镜检查

摘要目的探讨正常人共焦显微镜下角膜各层细胞的活体细胞形态学特征及密度.方法对28例(年龄36.6士14.5岁,范围19～68岁)41眼(男14眼,女27眼)正常人中央部角膜进行活体共焦显微镜检查,描述角膜各层结构的图象特点,并分析角膜各层细胞的密度与年龄的相关性.结果上皮表层细胞排列疏松,边界清楚,核反光较强;基底层细胞排列紧密,胞质和核反光弱,偶尔可见点状细胞核,细胞平均密度为5724.41士562.5个/mm2.Bowman's膜为无一定形状的膜状物,仅可分辨的结构是上皮下的念珠状神经纤维.基质层中可见相对较暗的背景下明亮的角膜细胞核.前基质内有放射状分布的神经纤维.前基质层角膜细胞平均密度为1099.10士164.90个/mm     

激光共焦显微镜对正常人眼角膜缘和中央角膜的观察

目的应用激光共焦显微镜对正常人眼角膜缘和中央角膜的组织结构与细胞形态的观察和分析。方法选择15名正常人的28只眼接受常规裂隙灯显微镜和检眼镜检查后,作为正常健康眼入选本研究。使用激光共焦显微镜对其上、下方角膜缘和角膜中央区进行检查,各层角膜图像均被记录,观察组织结构和细胞形态,对细胞密度进行计数并分析。结果所获角膜缘和角膜中央各层平面图像(x,y轴)及纵向断层图像(z轴)均非常清晰,同时获取动态录像。上、下方角膜缘均呈现Vogt栅栏状结构,并动态观察到血细胞在血管内的流动。表层上皮细胞排列非常疏松,边界明亮,胞体发暗,上方和下方角膜缘表层上皮细胞平均密度分别为(812±297)个/mm2和(785±263)个/mm2,二者比较差异无统计学意义(P>0.05)。上皮下可见明亮的Langerhans细胞,形态不规则,呈树枝状,上方和下方角膜缘Langerhans细胞平均密度分别为(288±102)个/mm2和(254±127)/mm2,二者比较差异无统计学意义(P>0.05)。角膜中央表层上皮细胞排列疏松,边界发亮,胞体发暗,细胞平均密度为(1098±315)个/mm2,多于上方和下方角膜缘(P<0.05)。基底上皮细胞排列紧密。上皮下和前基质层可见反光强烈的神经纤维丛,旁边偶见明亮的Langerhans细胞,形态不规则,细胞密度难以计算。浅层的神经纤维细小、弯曲度大、多小分支,深层的神经纤维粗大、弯曲度小、少见分支。基质层暗背景下散在分布细长的基质细胞,边缘欠清,细胞核明亮呈纺锤形。内皮细胞为排列整齐的六边形细胞,胞体发亮,边界发暗。角膜中央全层、基质层、上皮层厚度分别为(543.0±62.9)、(462·0±69.5)、(59.9±11.2)μm。结论激光共焦显微镜不仅可以对角膜进行无创的、实时的、活体的检查,而且与传统的光学共焦显微镜相比,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缘图像,对角膜疾病尤其是角膜缘疾病的基础研究与临床诊断将更有价值。     

In vivo confocal laser scanning microscopy of the cornea in dry eye

Background We carried out an investigation into the morphological and quantitative corneal properties in dry eye with various underlying pathologies.Methods Ten patients with aqueous tear deficiency, 8 with dysthyroid ophthalmopathy, 8 with chronic lagophthalmos and 10 normal participants were examined. Confocal microscope images were taken at the centre and at the lower and upper periphery of the cornea. Quantitative and morphological assessments of the epithelium, of the sub-basal nerves, of the stroma and the endothelium were made. The epithelial and corneal thicknesses were measured.Results The mean superficial and intermediate epithelial cell densities in the central cornea in the patient groups were significantly lower than in normal participants (p<0.01). The peripheral epithelial thickness was smaller (p<0.01); it was smallest in the lagophthalmos group. The cornea was thinner in the patient groups (p<0.01). For sub-basal nerves, the density had decreased (p<0.05), and in lagophthalmos the number of beadlike formations had increased (p<0.001); in some patients we found irregular branching patterns.Conclusions Dry eye patients showed significant alterations in the cornea, presumably due to increased desquamation of the superficial cell layer. This was most pronounced at the lower periphery of the cornea in patients with exposure keratopathy.The authors had no financial relationship with the organisation that supported the research.The authors had full control of all primary data and agree to allow Graefe’s Archive for Clinical and Experimental Ophthalmology to review the data if requested.The results were partly presented at DOG in 2005.     

正常中国汉族儿童角膜的共焦显微镜研究

目的采用角膜激光共焦显微镜对正常中国汉族儿童活体角膜各层组织结构进行观察,分析正常中国汉族儿童角膜各层细胞的活体细胞形态学特征及密度。方法 49例6～13岁正常中国汉族儿童中央部角膜进行活体共焦显微镜检查,研究角膜各层结构的图象特点,并分析角膜各层细胞的密度,与成年人进行对比。结果中国汉族儿童正常角膜上皮表层细胞排列疏松,边界清楚,胞体较大,核反光较强,又称翼状细胞;基底层细胞排列紧密,呈规则的蜂巢状,胞质反光弱,正常情况下未见细胞核,细胞平均密度为5947.58±769.3个/mm2。前弹力膜为无细胞结构的膜状物,可见串珠状的神经纤维走行。基质层中可见在相对较暗的背景下明亮的角膜基质细胞核。角膜基质内有神经纤维分布。前基质层角膜细胞平均密度为1621.12±123.26个/mm2,后基质层角膜细胞平均密度为984.71±113.17个/mm2,前后基质层细胞密度有显著性差别（P〈0.05）。后弹力层中无细胞结构,为均一无结构组织。内皮细胞层表现为规则的六边形结构,细胞结构清晰,细胞平均密度为3682.38±251.87个/mm2。左右眼及男女之间角膜各层细胞密度的差异无统计学意义（P=0.341～0.611和P=0.194～0.855）。各层角膜细胞的面积和密度与性别和眼别无差异。中国汉族儿童角膜各层细胞的密度较正常成人高。结论角膜激光共焦显微镜能够在实时、活体和三维空间从细胞水平清晰观察中国汉族儿童活体角膜各层细胞的形态结构,可以对角膜各层结构进行定性和定量分析,在儿童角膜病的研究和临床诊断方面是十分有用的工具。     

In vivo confocal microscopy in blepharitis

BACKGROUND: Dysfunction of the meibomian glands with inflammation and obstruction has been suggested to be an important factor in the pathogenesis of chronic blepharitis. Few objective tests are, however, available to examine the meibomian glands directly. PATIENTS AND METHODS: Nineteen patients with anterior blepharitis, meibomitis, meibomian gland dysfunction or severe keratoconjunctivitis sicca associated with blepharitis as well as 10 patients with normal lid margins were examined with the HRTII/RCM in vivo confocal microscope. Scans of the tear film, the tarsal conjunctiva, the hair follicles and the meibomian glands were analysed by a masked observer. RESULTS: Patients with normal lid margins exhibited a minimal round cell infiltrate in the tarsal conjunctival epithelium and largely normal ducts of the meibomian glands lined with a multilayered epithelium as well as normal gland acini. In patients with anterior blepharitis, blepharitis associated with autoimmune peripheral ulcerative keratitis and blepharitis in the context of severe dry eye, confocal microscopy disclosed normal meibomian glands. In 12 patients with blepharitis/meibomitis or meibomian gland dysfunction, profound pathology was visible with dilatation and obstruction of the meibomian gland ducts. In 15 of 19 patients with blepharitis/meibomitis, but not in meibomian gland dysfunction, an intense inflammation was observed in the tarsal conjunctival epithelium and stroma. In one patient, demodex folliculorum was evident in vivo. In patients with normal lid margins as well as in patients with blepharitis, hair follicles appeared within normal limits. CONCLUSIONS: In vivo confocal microscopy allowed the examination of the tear film, the tarsal conjunctiva, the lid margin including the lash follicles and the meibomian glands. In patients with meibomian gland disease pathological changes could be visualised and documented objectively. The presence of an inflammatory infiltrate permitted us to differentiate between meibomitis and meibomian gland dysfunction. Changes of the lash follicles do not seem to play an important role in blepharitis. Thus, in vivo confocal microscopy represents an objective technique in the classification and follow-up of patients with blepharitis.