血管生成拟态与翼状胬肉进行期及静止期的相关性研究

目的：研究血管生成拟态与翼状胬肉发生发展的相关性,初步探讨其临床意义。
　　方法：选取20例正常眼的结膜组织及翼状胬肉静止期和进行期各50例标本行苏木素－伊红（ HE）染色、过碘酸雪夫（ PAS）染色、免疫组化CD34、VEGF染色及免疫组化CD31染色联合PAS双重染色观察翼状胬肉新生血管和血管生成拟态情况。
　　结果：CD34和VEGF在翼状胬肉组织中的表达均高于正常结膜组织（两者P＜0．05）,并且两者在进行期翼状胬肉组织中的表达均高于静止期（两者P＜0．05）。 PAS染色血管拟态区网眼间隔染成紫红色,进行期和静止期出现血管生成拟态的例数分别是38例和11例,差异具有统计学意义（P＜0．05）,相关性分析显示血管生成拟态与进行期翼状胬肉呈显著密切正相关（ Spearman’s相关系数r＝0．540＞0．5,P＝0．000）。
　　结论：翼状胬肉除了新生小血管形成增多外,血管生成拟态也是其血供方式之一,并且血管生成拟态与翼状胬肉的进展具有密切关系。     

CD34在翼状胬肉中表达的意义

目的研究翼状胬肉组织中CD34的表达及意义。方法对50例手术切除的翼状胬肉标本行苏木精-伊红染色、过碘酸希夫染色，并进行形态学、免疫组织化学研究和免疫荧光共聚焦显微镜下观察CD34、波形蛋白（VIM）、血管内皮生长因子（VEGF）在翼状胬肉中的表达。结果在翼状胬肉标本中，CD34在血管内皮细胞、血管周细胞以及一些散在的星形、梭形细胞中表达阳性，而在血管壁上及成熟的胶原细胞上呈阴性表达；VIM阳性表达于大部分翼状胬肉组织中；组织中部分疏松排列的网状及裂隙样、管样区结构过碘酸希夫染色呈阳性的紫红色反应；网状区的梭形细胞VEGF阳性表达。结论翼状胬肉组织中CD34阳性细胞来源于间充质干细胞的分化。翼状胬肉除以血管发生和血管新生的方式生成血管外，血管拟态可能是另外一种供血途径。     

CD44、SOX9、MMP9、Vimentin在翼状胬肉中的表达及其意义

目的 研究CD44、SOX9、基质金属蛋白酶9(matrix metalloproteinase,MMP9)及波形蛋白(Vimentin)在原发性翼状胬肉组织中的表达及其意义.方法 对25例原发性胬肉组织和5例正常结膜组织的组织切片用CD44、SOX9、MMP9及Vimentin蛋白的抗体进行免疫组织化学染色,观察上述蛋白在翼状胬肉组织和正常结膜组织组织内的表达,对结膜上皮内CD44和SOX9染色阳性的细胞进行计数,并比较翼状胬肉组织与正常结膜组织内的细胞阳性率.结果 HE染色结果显示,正常结膜和翼状胬肉组织均由上皮层和固有层组成;正常结膜的上皮层次较少,固有层含有少量血管;翼状胬肉的上皮细胞层次明显增多,固有层内含有密集分布的血管.免疫组织化学染色阳性细胞计数的统计学数据表明,正常结膜组织内CD44阳性细胞率为15％±3％,翼状胬肉组织内为70％±15％;正常结膜组织内SOX9阳性细胞率为17％±4％,翼状胬肉组织内为75％±13％;两组的细胞阳性率差异均有显著统计学意义(均为P＜0.001).MMP9及Vimentin阳性细胞广泛分布于翼状胬肉组织的上皮及上皮下基质层内,在正常结膜组织内阳性细胞呈散在分布,翼状胬肉组织的免疫组织化学染色强度高于正常结膜组织.结论 CD44、SOX9、MMP9及Vimentin在翼状胬肉组织中高表达,可能在翼状胬肉的发生、发展中起着重要作用.     

血管生成拟态在翼状胬肉进展中的作用及可能机制

目的　研究血管生成拟态（VM）在翼状胬肉进展中的作用及其与基质金属蛋白酶-2（MMP-2）、基质金属蛋白酶-9（MMP-9）和血管内皮生长因子（VEGF）表达的关系,探讨VM与翼状胬肉进展的相关性及可能机制。方法　筛选符合条件的翼状胬肉标本,进行胬肉分期（静止期/进展期）、分型（初发型/复发型）及临床特征指标采集。采用CD31/PAS双重染色检测不同类型翼状胬肉中VM的形成差异。以初发型进展期翼状胬肉为研究对象,HE染色观察病理特征,免疫组织化学染色及Western blot检测MMP-2、MMP-9及VEGF的表达,分析VM与初发型进展期翼状胬肉临床病理特征及MMP-2、MMP-9、VEGF的相关性。结果　静止期翼状胬肉VM阳性率为15.38%,进展期翼状胬肉VM阳性率为68.50%,组间差异具有统计学意义（P=0.000）;初发型翼状胬肉VM阳性率为40.80%,复发型翼状胬肉VM阳性率为81.48%,组间差异亦具有统计学意义（P=0.000）。VM与进展期及复发型翼状胬肉均呈正相关（均为P<0.05)。VM阳性的初发型进展期翼状胬肉组织,其血管分级、上皮细胞层数、杯状细胞数、成纤维细胞数、血管数、炎症细胞数、纤维化程度、MMP-2、MMP-9及VEGF的表达均明显高于VM阴性者,而变性程度低于VM阴性者,差异均有统计学意义（均为P<0.05）。初发型进展期翼状胬肉VM与翼状胬肉血管分级、上皮细胞层数、杯状细胞数、成纤维细胞数、血管数、炎症细胞数、纤维化程度、MMP-2、MMP-9及VEGF的表达均呈正相关,与胬肉变性程度呈负相关（均为P<0.05)。结论　翼状胬肉组织中存在VM结构,可作为其血供途径之一,VM可促进翼状胬肉的进展,MMP-2、MMP-9及VEGF可能是翼状胬肉VM形成的分子机制。     

翼状胬肉组织中CD34、AC133、STRO-1、C—KIT的表达

目的评估源白骨髓的干细胞与翼状胬肉发病的相关性,探寻翼状胬肉的发病机制。方法选择翼状胬肉头部达到瞳孔缘的病例12例（12眼）,其中男6例,女6例;年龄51～66岁;原发性翼状胬肉8例,复发性翼状胬肉4例,均长于鼻侧。对照组材料来自同一眼球、距翼状胬肉边缘约10mm的正常球结膜组织。采用EnVision免疫组织化学法检测骨髓干细胞标志物来源的CD34、AC133、STRO-1、C-KIT的表达。结果光镜下可见翼状胬肉组织中有大量带有骨髓干细胞或祖细胞阳性标志物来源的细胞,翼状胬肉组织头部阳性表达更强。4种阳性细胞的分布及细胞形态类似。C—KIT阳性表达细胞聚集于翼状胬肉上皮基底组织和基质组织。上皮基底部阳性细胞呈圆形或椭圆形,而在基质组织中呈纺锤形且主要分布于血管周围。CD34阳性表达细胞主要分布于翼状胬肉组织上皮基底层,在血管内皮也有阳性表达,类似于C—KIT,但比C—KIT上皮层组织阳性细胞少。细胞形态在上皮基底部呈圆形或椭圆形。AC133阳性表达细胞主要分布于上皮层,基质和血管内皮中也有阳性表达,形态类似CD34和C-KIT。STRO-1阳性表达细胞的分布不同,主要见于基质层,阳性细胞形态呈纺锤形,而上皮层中未见表达。各种因子的表达在原发性和复发性翼状胬肉间无明显差别。对照组切片中标志物的表达均呈阴性。结论带有源白骨髓干细胞阳性标志物的细胞在翼状胬肉的发病和术后复发过程中可能起重要作用。     

激光共焦显微镜与组织病理学对翼状胬肉组织中微血管密度研究的比较

背景 研究表明,翼状胬肉是眼表的一种异常细胞增生性疾病,血管因素在它的生长和复发中起重要作用.多种抗新生血管治疗方法被应用于翼状胬肉,但活体观察翼状胬肉血管情况的研究尚少见.目的 比较角膜激光共焦显微镜与免疫组织化学染色对翼状胬肉组织中微血管密度研究的结果. 方法 采用前瞻性病例对照研究方法,收集原发性翼状胬肉患者20例20眼为翼状胬肉组,手术中获取翼状胬肉组织;另收集年龄和性别匹配的行内眼手术及斜视手术的患者20例20眼为正常球结膜组,手术中获取正常鼻侧球结膜组织作为对照,翼状胬肉组患者均于术前用海德堡角膜激光共焦显微镜采集5幅翼状胬肉头部的微血管图像,同时采集正常球结膜组正常球结膜微血管图像.对两组患者手术中获得的标本进行常规组织病理学检查和CD31免疫组织化学染色,计算组织标本中微血管密度值,对翼状胬肉组患者激光共焦显微镜所测的微血管密度值以及手术标本中的微血管密度值分别与正常球结膜组进行比较,并分别将两组用两种方法测得的血管密度值进行相关分析.结果 角膜激光共焦显微镜下翼状胬肉基质中的微血管密度( MVD)值为( 8929±2993) μm/mm2,正常球结膜中的微血管密度值为( 4202±692) μm/mm2,二者比较差异有统计学意义(t=6.881,P＜0.01).翼状胬肉的组织标本中CD31染色阳性的微血管数为(21.00±4.06)个/高倍视野,正常球结膜组为(6.07±1.75)个/高倍视野,二者比较差异有统计学意义(t=12.312,P＜0.01).翼状胬肉组和正常球结膜组激光共焦显微镜活体测得的MVD值与免疫组织化学离体测得的MVD值均呈显著正相关(翼状胬肉组:r=0.649,P＜0.01;正常球结膜组:r=0.572,P＜0.01).结论 角膜激光共焦显微镜活体动态观察翼状胬肉组织微血管的方法优于离体的免疫组织化学检测法,角膜激光共焦显微镜对翼状胬肉的血管形态进行活体研究有助于进一步研究翼状胬肉组织的生物学行为及其机制.     

基质细胞衍生因子及其受体CXCR4在翼状胬肉的异常表达

目的探讨基质细胞衍生因子(stromal cell-derived factor-1,SDF-1)及其受体CXCR4在翼状胬肉的表达及意义。方法用免疫组织化学染色方法对4例正常人结膜组织、11例初发性翼状胬肉和9例复发性翼状胬肉组织中SDF-1/CXCR4蛋白的表达进行检测。结果在正常结膜组织中SDF-1/CXCR4仅表达于基底部上皮细胞;胬肉组织中有17例,全层上皮细胞均有SDF-1/CXCR4的表达,胬肉基质层中浸润的炎性细胞、血管内皮细胞均有CXCR4的蛋白表达;3例初发性翼状胬肉表达方式与正常结膜相似。结论翼状胬肉组织SDF-1及其受体CXCR4蛋白表达增加;上皮细胞分泌的SDF-1通过与其受体CXCR4的相互作用可能参与了翼状胬肉的血管形成和炎性细胞浸润。     

多西环素抑制视网膜母细胞瘤血管生成拟态的体外研究及组织学观察

研究多西环素对人视网膜母细胞瘤(RB)细胞体外血管生成拟态 （VM）和细胞增生的抑制效应。方法建立人RB细胞株三维培养系统,缺氧诱导后观察RB细胞能否形成血管网状结构及其特点;用5～20 mg/L不同浓度多西环素处理RB细胞, 过碘酸雪夫氏(PAS)染色观察对RB细胞体外血管生成拟态的抑制效应;噻唑蓝(MTT)比色法检测多西环素对RB细胞增生活性的变化; 逆转录聚合酶链式反应（RT-PCR）技术检测不同条件下基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)mRNA的相对表达量; 收集12例RB石蜡包埋样本进行CD34/PAS免疫组织化学双重染色,观察RB中的血管生成拟态。结果 RB细胞在三维培养系统形成管网状的拟态血管; 5～20 mg/L多西环素能抑制RB细胞体外管状结构数量。MTT检测显示多西环素加药组3、5、7 d的吸光度A［旧称光密度（OD）］值与对照组比较差异均有统计学意义(t=15.320,P <0.01),且不同浓度的多西环素组之间细胞的增生抑制率差异亦有统计学意义(F=9.442,11.729,12.120;P <0.05),其浓度与细胞的增生呈负相关(r=-0.924, P <0.01), 氯化钴缺氧组MMP-2、MMP-9的mRNA表达量明显增强,各浓度多西环素加药组显著低于氯化钴缺氧组(t=16.469,P <0.01)。在12例RB组织标本中可见CD34阴性PAS阳性的瘤细胞围成管道样结构,部分管腔中可见红细胞。结论 多西环素能够抑制RB体外血管生成拟态形成和细胞的增生,其机制可能是通过抑制MMP-2、MMP-9 mRNA的表达而实现。     

翼状胬肉组织中环氧化酶-2及p21-ras的表达研究

目的 通过免疫组织化学方法检测翼状胬肉组织中环氧化酶-2（COX-2）和p21ras的表达，进一步证实氧自由基在翼状胬肉形成和发展中的作用。方法 选择临床切除的静止期、进行期和复发期胬肉材料共40例，行苏木精-伊红染色及COX-2和p21ras免疫组织化学染色。结果 COX-2及p21ras在正常球结膜中表达微弱，静止期胬肉组织中COX-2和p21ras阳性染色主要位于复层鳞状上皮化生细胞，在进行期及复发期翼状胬肉组织中COX-2及p21ras阳性染色弥漫性分布于复层鳞状上皮化生细胞的胞浆和杯状细胞的胞浆中，胬肉组织中的炎性细胞和血管内皮细胞胞浆可见少量COX-2及p21ras表达。结论 p21ras and COX-2的过度表达在翼状胬肉的发生和发展过程中可能起重要作用。     

翼状胬肉组织病理学研究及相关因子的检测

目的：研究翼状胬肉组织病理学特征并检测翼状胬肉组织中碱性成纤维细胞生长因子（ｂＦＧＦ）、转化生长因子－β１（ＴＦＧ－β１）的表达,探讨这些因子对翼状胬肉的促增殖作用,为临床治疗提供有益的启示。方法：以２３例正常结膜组织为对照,行５６例翼状胬肉组织石蜡切片,Ｈ－Ｅ染色、Ｍａｓｓｏｎ’ｓ三色法染色以及免疫组织化学ＳＰ法染色,观察翼状胬肉内各种组织病理改变,并检测翼状胬肉组织中相关因子ｂＦＧＦ、ＴＧＦ－β１的表达情况。结果：翼状胬肉上皮以增生表现为主,部分静止期胬肉上皮变薄;基质层可有增生和变性两种改变：基质浅层多为增生性病变,并有大量新生血管和炎性细胞浸润。基质深层主要为胶原纤维变性,１８例静止期胬肉病理学特征以变性为主;３２例进行期胬肉同时存在增生和变性改变;６例复发型胬肉则以增生为其主要病理表现。ｂＦＧＦ及ＴＧ     

Angiogenesis in pterygium: morphometric and immunohistochemical study

OBJECTIVE: To evaluate the role of angiogenesis in the pathogenesis of pterygium by comparing the expression of von-Willebrand factor (vWF) and vascular endothelial growth factor (VEGF) in pterygium, and in normal superior bulbar conjunctiva. METHODS: 23 human samples from pterygium and the superior bulbar conjunctiva were stained using rabbit anti-vWF and anti-VEGF antibodies. The density of vWF and VEGF positive vessels, VEGF staining intensity and the number of VEGF positive stromal, epithelial and vascular endothelial cells were evaluated. RESULTS: Pterygium specimens had higher average vWF and VEGF positive microvascular counts per high power field (P = 0.0012), higher average VEGF staining intensity scores in epithelial, stromal and endothelial cells (p < 0.0001) and higher VEGF positive cell counts (P < 0.0001) than normal conjuctiva. CONCLUSIONS: Over-expression of VEGF in pterygium tissue, together with the abundance of vWF-stained new vessels, may support previous suggestions that angiogenesis may play a role in the formation of pterygium.     

Investigation of vasculogenic mimicry in sebaceous carcinoma of the eyelid

Purpose: Vasculogenic mimicry (VM) is a newly proposed pattern of tumour angiogenesis that has been identified in some malignancies and is associated with poor prognosis. The purpose of this study was to investigate whether sebaceous carcinomas of the eyelid exhibit VM and to determine whether these fluid‐conducting patterns are associated with clinicopathologic features, the number of microvessels and the levels of endothelial growth factor (VEGF) and matrix metalloprotease‐2 (MMP‐2) in tumours. Methods: Forty paraffin‐embedded samples of sebaceous carcinoma of the eyelid were collected, along with complete clinical and pathologic data for all the cases. Tissue sections were stained for CD34, periodic acid and Schiff (PAS), VEGF and MMP‐2. VM was identified by the presence of PAS‐positive and CD34‐negative loops lined by tumour cells. The VM status of tumour samples was compared with the clinical and pathological data using statistical tests. The levels of VEGF, MMP‐2 and the number of microvessels were compared between patients with and without VM. Results: VM was detected in 14 of 40 (35%) tumour samples. The existence of VM in tumours was associated with tumour size (p = 0.007) and recurrence (p = 0.021). The number of microvessels was lower in tumours with VM (13.03 ± 4.02 versus 22.99 ± 7.72; p < 0.0001). The staining index of MMP‐2 was higher in tumours with VM (27.43, range: 0–5.3) compared to tumours without VM (16.77, range: 0–2.7; p = 0.004). However, there was no difference in the expression of VEGF between groups with and without VM (p = 0.244). Conclusions: Vasculogenic mimicry is present in sebaceous carcinoma of the eyelid making it an unfavourable prognosis sign. MMP‐2 is associated with VM formation in sebaceous carcinoma of the eyelid.     

多西环素对人翼状胬肉成纤维细胞血管生成拟态形成的影响

目的：探讨多西环素(DOX)对人翼状胬肉成纤维细胞(HPFs)血管生成拟态(VM)形成的影响及其可能的分子机制。

方法：采用波形蛋白(Vimentin)和角蛋白(CK)免疫细胞化学染色鉴定原代培养的HPFs。分别设置对照组、DOX低、中、高浓度(50、100、200mg/L)组,应用CCK-8法和划痕实验检测各组HPFs活力及迁移能力,利用细胞三维培养及过碘酸-雪夫(PAS)染色观察各组细胞VM生成情况,并通过Western blot法检测各组细胞基质金属蛋白酶(MMP)-9及血管内皮生长因子(VEGF)的表达情况。

结果：免疫细胞化学染色结果显示,细胞呈梭形且Vimentin阳性表达,CK阴性表达,符合成纤维细胞特性。与对照组相比,DOX组细胞活力、迁移能力、VM密度及MMP-9和VEGF蛋白的表达均显著降低(P<0.05),DOX各浓度组组间相比均有差异(P<0.05)。相关性分析提示HPFs形成的VM密度与MMP-9和VEGF蛋白表达水平呈显著正相关(r=0.949、0.960,均P<0.05)。

结论：DOX可通过减少MMP-9及VEGF的表达抑制HPFs的活力及迁移能力,减少VM的形成,提示MMP-9和VEGF可能是翼状胬肉形成VM的分子机制。     

Distinctive distribution of HLA class II presenting and bone marrow derived cells in the anterior segment of human eyes.

Cells of bone marrow origin that normally occupy the stroma of the murine iris and ciliary body have been implicated in the immune phenomenon, anterior chamber associated immune deviation (ACAID). Following injection of antigen into the anterior chamber, cells of this type deliver an ACAID inducing signal into the systemic circulation, presumably through the outflow tract. In an effort to identify such cells in man, anterior chambers of 34 human donor eyes of different age groups were stained immunocytochemically with monoclonal antibodies directed at HLA class II molecules, CD 45 (a molecular marker of bone marrow-derived cells) and macrophage-associated membrane molecules (CD 68, CD 14). Within the outflow tissue, the cells of the filtering trabecular meshwork stained with none of those reagents. However, infrequent single, dispersed, dendritic cells were positively stained in the intertrabecular spaces. More numerous labelled cells were found in the anterior- and posterior-most portions of the non-filtering part of the trabecular meshwork. These cells were continuous with stained cells adjacent to the outer wall of Schlemm's canal and to the collector channels. Numerous labelled cells were seen in the vicinity of the intra- and episcleral vessels, the ciliary meshwork, the stroma of the ciliary muscle and epithelial processes, and the iris stroma. With advancing age, increasing numbers of CD 45+, HLA class II expressing cells appeared to accumulate in the so-called uveoscleral pathway. These results indicate that bone marrow-derived cells with the potential to function of ACAID induction reside within human eyes, and that cells of this type are located not only in the stroma of iris and ciliary body, but within the non-filtering portions of the trabecular meshwork and the uveoscleral pathway. The appearance of rare CD 45+ cells "in transit" in the filtering trabecular meshwork is compatible with the view that cells carrying ACAID-inducing signals to the systemic immune apparatus escape from the eye by this route.     

Epithelial metaplasia of the corneal endothelium in Fuchs endothelial dystrophy

PURPOSE: To evaluate the immunohistochemical characteristics of human corneas with the diagnosis of Fuchs endothelial dystrophy (FED). METHODS: Formalin-fixed, paraffin-embedded sections of corneas with the diagnosis of FED (15 patients) and 10 control corneas were stained with hematoxylin-eosin and periodic acid-Schiff (PAS). Adjacent histologic sections were stained with monoclonal antibodies that react with epithelial antigens: pancytokeratin, cytokeratins (CK) 7 and 20 CAM 5.2, epithelial membrane antigen (EMA), and Ber EP4. Eight corneas were stained with antibodies to vimentin, smooth-muscle actin (SMA), and CD 68. RESULTS: The endothelial cells in FED were attenuated and atrophic; some contained pigment consistent with melanin. The endothelial cells stained for pancytokeratin, CK 7, and vimentin in all corneas of FED, whereas variable staining was noted with CAM 5.2. No staining of endothelium was noted with CK 20, EMA, BerEP4, SMA, or CD 68. CONCLUSION: Some cytokeratins that are normally restricted to true epithelium are present in the endothelium of FED. Epithelial metaplasia of endothelium in FED may represent a nonspecific response of distressed endothelial cells, as previously reported in posterior polymorphous dystrophy, congenital hereditary endothelial dystrophy, and iridocorneal endothelial syndrome.     

视网膜母细胞瘤血管生成拟态与临床病理学特征关系的研究

目的 探讨视网膜母细胞瘤(Rb)标本中是否存在血管生成拟态(VM),并分析VM与Rb临床病理特征及其与缺氧诱导因子-1αa(HIF-1α)、血管内皮生长因子(VEGF)蛋白表达的关系,进一步阐述VM的临床意义.方法 实验研究.收集60例Rb患者、10例正常人视网膜组织石蜡标本及相关临床病理资料,应用过碘酸-雪夫(PAS)与CD34双重染色以及免疫组织化学染色法检测60例Rb中是否存在VM和分布,观察与临床病理学特征的关系特点;检测HIF-1α、VEGF蛋白表达,分析Rb瘤组织中VM与HIF-1α、VEGF表达的关系;用CD34抗体进行肿瘤血管内皮染色,并计数肿瘤微血管密度(MVD).Rb和正常视网膜组织中的VM、HIF-1α、VEGF蛋白阳性表达的比较采用χ2检验;VM阳性组和VM阴性组的MVD值的比较采用q检验.结果 HE染色法观察到Rb中存在VM,其为Rb细胞构成的管腔样结构,无内皮细胞衬覆,腔内可见红细胞.CD34和PAS双重染色结合CD34、神经元特异性烯醇化酶(NSE)免疫组织化学染色进一步证明Rb中存在VM,60例Rb中VM阳性率为18.33%(11/60),其中随着R-E分级的增高,VM阳性表达率明显增加,差别具有统计学意义(χ2=8.861,P<0.05);分化型Rb中VM阳性率(4.34%)低于未分化型中VM阳性率(22.02%)(χ2=4.872,P<0.05);此外,HIF-1α、VEGF在Rb的VM形成过程中的阳性表达率依次为VM阳性组大于VM阴性组大于正常组;微血管密度计数高低与VM的表达有关,VM阴性组的MVD值(49.77±2.05)高于VM阳性组MVD值(36.53±1.15).结论 Rb中存在VM,且肿瘤恶性程度越高,形成VM能力越强.存在VM的Rb组织HIF-1α、VEGF高表达,提示HIF-1α、VEGF在VM的形成中可能起促进作用.     

Connective tissue growth factor in pterygium: simultaneous presence with vascular endothelial growth factor – possible contributing factor to conjunctival scarring

BACKGROUND: Various growth factors have been detected in pterygium and been associated with its vasculogenesis. The basic pathophysiological mechanisms responsible especially for the fibrotic activity in pterygium are, however, not yet known. Connective tissue growth factor (CTGF) has been shown to be substantially involved in various processes of fibrosis. We report on the presence of CTGF in pterygium and its simultaneous presence with vascular endothelial growth factor (VEGF). METHODS: Samples of pterygium were collected during surgery with informed consent of the patients. Specific, non-commercial primary antibodies against CTGF were used to detect CTGF using immunohistochemistry. Specificity of antibodies was confirmed with Western-blot analysis. The same specimens were stained with commercial antibodies for VEGF. Additionally RT-PCR analysis was performed from pterygium samples. RESULTS: CTGF was detected in the epithelium of all samples as well as in some stromal keratocytes. The RT-PCR confirmed the identity of CTGF in these samples. The staining pattern differed slightly from that of VEGF, which was detected in all samples. The control sections were negative. CONCLUSION: CTGF is present in the epithelium of a majority of pterygia and probably contributes to fibrosis. Simultaneous presence with VEGF suggests growth factor interaction and possible involvement in apoptotic dysregulation.