红斑狼疮皮损内角质形成细胞凋亡和SSA/Ro抗原的表达

目的：探索光敏感红斑狼疮(LE)皮损形成的机理和SSA／Ro抗原表达的来源。方法：用原位切口标记法和免疫荧光法测定了19例光敏感型红斑狼疮(LE)患新鲜皮损内自身抗原SSA／Ro表达情况和角质形成细胞凋亡情况。结果：发现7例LE皮损表皮内棘细胞周围有不同程度的颗粒状SSA／Ro抗原表达；而对照组无1例荧光着色，13例皮损表皮棘细胞内可见凋亡细胞荧光着色。结论：光敏感LE患的角质形成细胞通过细胞凋亡释放SSA／Ro抗原，并参与皮损形成。     

尖锐湿疣皮损角质形成细胞ICAM-1及HLA-DR表达的研究

目的:研究细胞间黏附分子-1(ICAM-1)及人白细胞分化抗原DR区(HLA-DR抗原)在尖锐湿疣(CA)发病中的作用。方法:采用免疫组织化学技术一链酶亲和素一生物素复合物法(SABC法)对比研究CA初发组、CA复发组及正常人角质形成细胞(KC)表面ICAM-1及HLA-DR抗原的表达。结果:CA初发组与复发组间ICAM-1及HLA-DR抗原表达无显著性差异(P>0.05)。病例组中KC表面ICAM-1及HLA-DR抗原表达有一定相关性(r=0.4316,P<0.05)。结论:CA患者病损处KC表面表达ICAM-1及HLA-DR抗原使局部细胞免疫状态发生变化,诱导T细胞在病损处聚集、活化,是有效地启动并参与局部免疫应答、促进CA病损消退的关键。     

尖锐湿疣损害中角质形成细胞凋亡与增殖细胞核抗原表达的关系

我们用ABC免疫组化技术和TUNEL原位标记法分别检测了尖锐湿疣（CA）损害中角质形成细胞增殖细胞核抗原（PCNA）的表达及细胞凋亡的变化，旨在探讨人乳头瘤病毒（HPV）引起上皮增殖的机理。一、材料与方法５１例CA标本来自我院１９９８年３月至２００１年１月存档的石蜡包埋组织，其中男２１例，女３０例，年龄１９～６０岁（平均３５．９岁），病程７d至７个月（平均５８d），全部病例均有典型的临床表现及组织病理变化。对照组１８例，其中男８例，女１０例，年龄１８～５８岁（平均３２岁），标本为手术切除的正常包皮或阴道粘膜…     

尖锐湿疣皮损角质形成细胞中β-catenin的表达

目的探讨β-catenin与尖锐湿疣皮损角质形成细胞增生的关系。方法采用聚合酶链反应(PCR)方法检测尖锐湿疣皮损感染的HPV分型,将HPV16/18型皮损分为高危型组(CA1),HPV6/11型皮损分为低危型组(CA2),每组30例;分别应用免疫组织化学及逆转录聚合酶链(RT-PCR)方法检测β-catenin蛋白及β-catenin mRNA在CA1组和CA2组角质形成细胞中的表达。结果在正常皮肤组织中β-catenin蛋白阳性物质均表达于胞膜;CA1组和CA2组中主要表达于胞核和胞质,CA1组和CA2组角质形成细胞中β-catenin蛋白均呈异常表达,异常表达率(分别为86.70%,40.00%)均显著高于正常对照组(0);CA1组和CA2组角质形成细胞中β-catenin mRNA的表达水平(阳性分别为23例,18例)亦均显著高于正常对照组(阳性10例)。结论尖锐湿疣皮损角质形成细胞中β-catenin表达异常,在胞膜、胞浆及胞核的分布与正常角质形成细胞不同,可能与细胞过度增殖有关。     

皮肤型红斑狼疮皮损中SASPase的表达和意义

目的 探讨SASPase在皮肤型红斑狼疮皮损中的表达水平及其在发病机制中的意义.方法 无血清培养原代角质形成细胞,将提取的蛋白样品采用固相pH梯度双向凝胶电泳进行分离,应用ImageMaster 2D Platinum 5.0软件对图像进行匹配分析,选取差异表达蛋白质点,经基质辅助激光解吸附飞行时间质谱进行质谱鉴定.并通过免疫印迹验证表达水平.结果 成功培养角质形成细胞,获得重复性较好的双向电泳图谱,匹配点数在1200个左右,匹配率＞80％.鉴定SASPase在CLE皮损角质形成细胞表达升高,免疫印迹验证与双向电泳结果一致.结论 皮肤型红斑狼疮皮损的发生发展可能与SASPase的异常激活和过度表达有关.     

γ—干扰素诱导角质形成细胞凋亡及其Fas抗原表达研究

目的：探讨γ－干扰素在银屑病发病中的作用。方法：流式细胞仪测定亚二倍体含量、片段化DNA分析、Annexin V凋亡检测法、免疫组织化学、间接免疫荧光流式细胞仪测定Fas抗原表达。结果：γ－干扰素上调角质形成细胞Fas抗原表达（P＜0．01）,诱导角质形成细胞凋亡（P＜0．01）,结论：γ－干扰素可能通过上调角质形成细胞Fas抗原表达,进而诱导角质形成细胞凋亡而参与银屑病发病。     

γ—干扰素诱导角质形成细胞凋亡及其Fas抗原表达研究

目的：探讨γ－干扰素在银屑病发病中的作用。方法 通过流式细胞仪测定角持形成细胞中亚二倍体细胞含量、片段化DNA分析及AnnexinV法检测经γ－干扰素诱导的角质形成细胞凋亡;采用组织化学、流式细胞仪测定经γ－干扰素作用后的角质形成细胞Fas抗原表达。结果 γ－干扰素上调角质形成细胞Fas抗原表达（P＜0．01）,诱导角质形成细胞凋亡（P＜0．01）。结论 γ－干扰素可能通过上调角质形成细胞Fas抗原表达,进而诱导角质形成细胞凋亡而参与银屑病发病。     

他扎罗汀联合干扰素γ对角质形成细胞HLA-DR表达的影响

目的探讨他扎罗汀对角质形成细胞(KC)增殖及他扎罗汀、干扰素(IFN)-γ及二者联合对HLA-DR在KC上表达的影响,为他扎罗汀的抗增殖作用及其对炎症的影响进行初探。方法①体外培养正常人KC;②用MTT法检测他扎罗汀对KC增殖的影响;③用免疫组化的方法,分别观察他扎罗汀组、IFN-γ组及二者联合组对HLA-DR在KC上表达的影响。结果①10-7~10-5mol/L他扎罗汀作用KC24h、48h后,处理组的细胞增殖较对照组显著降低且呈剂量依赖性;②正常人KC基本不表达HLA-DR;③10-6mol/L他扎罗汀作用24h后,不能诱导KC表达HLA-DR;④500U/mLIFN-γ处理24h后可明显诱导KC表达HLA-DR;⑤10-7~10-5mol/L他扎罗汀联合IFN-γ组作用24h后,KC上HLA-DR的表达诱导作用较单独IFN-γ组显著增强(P<0.005),且呈剂量依赖性。结论他扎罗汀对体外培养的KC增殖有抑制作用;他扎罗汀对IFN-γ诱导KC表达HLA-DR有增强作用。     

角质形成细胞生长因子与银屑病的关系

角质形成细胞生长因子是近年来发现的有着重要生物功能的生长因子,它属于成纤维细胞生长因子家族,由成纤维细胞产生。角质形成细胞生长因子可刺激角质形成细胞的增生、分化、移行,促进上皮细胞的再生、增厚,对银屑病的发病机制及其治疗可能有一定的启示。本文就角质形成细胞生长长因子和的生物学功能及其与角质形成细胞及银屑病的关系做一定综述。     

一氧化氮、角质形成细胞和银屑病

一氧化氮是一种多效性生物气体分子，在皮肤组织的生理和病理过程中均具有重要作用。银屑病皮损区释放的一氧化氮量和诱导型一氧化氮合酶的表达均高于非皮损区和正常皮肤，其在银屑病中的作用机理不清楚。现综述一氧化氮在角质形成细胞凋亡、增生分化和基因表达调控的研究进展及其在银屑病中的可能作用机制。     

蕈样肉芽肿患者外周血淋巴细胞HLA-DR抗原表达的研究

目的探讨蕈样肉芽肿（ＭＦ）患者外周血淋巴细胞的ＨＬＡ ＤＲ抗原表达及其意义。方法应用流式细胞术及免疫双荧光染色法,共检测１０例ＭＦ。结果ＭＦ患者外周血ＨＬＡ ＤＲ＋淋巴细胞数及ＣＤ＋３ ＨＬＡ ＤＲ＋淋巴细胞数与正常对照的差异均有显著性意义（Ｐ分别＜０．０１,＜０．００１）,ＨＬＡ ＤＲ＋淋巴细胞数及ＣＤ＋３ ＨＬＡ ＤＲ＋淋巴细胞数与疾病临床分期均呈显著正相关（ｒ值分别为０．７９６、０．７６７,Ｐ均＜０．０１）。ＣＤ＋４ ＨＬＡ ＤＲ＋Ｔ细胞、ＣＤ＋８ ＨＬＡ ＤＲ＋Ｔ细胞均高于正常对照,差异非常显著（Ｐ分别＜０．０５、＜０．００１）。结论ＭＦ患者外周血淋巴细胞ＨＬＡ ＤＲ抗原表达普遍存在异常,与ＭＦ的发病密切相关。     

体外培养人黑素细胞HLA—DR抗原的表达

为了解正常人表皮黑素细胞是否表达ＨＬＡ－ＤＲ抗原,黑素细胞经体外培养后,是否因细胞增减的添加或培养时间的长短而影响ＬＨＡ－ＤＲ抗原表达而应用免疫酶染色法进行检测。结果发现表皮基底层的黑素细胞不表达ＨＬＡ－ＤＲ抗原;接种于含ＴＰＡ增减液的黑素细胞,在传代后２周,黑素细胞表达ＨＬＡ－ＤＲ抗原,７周后ＨＬＡ－ＤＲ抗原失表达;接种于不含ＴＰＡ培养液的黑素细胞,在传代后２周和７周时均不表达ＨＬＡ－ＤＲ抗原是     

HLA—DR抗原在斑秃皮损中的表达

目的：探讨斑秃的发病机理。方法：应用免疫组化的方法（SP法）观察了HLA－DR抗原在15例斑秃皮损中及8例正常头皮组织中的表达情况。结果：HLA－DR抗原在斑秃皮损毛囊角度形成细胞中明显表达，占60％（9／15）（P＜0．05）；HLA－DR抗原在正常头皮中未见表示。结论：HLA－DR抗原参与了斑秃发病，免疫功能紊乱可能是斑秃的发病因素。     

红斑狼疮患者皮损角朊细胞中雌激素受体检测结果浅析

应用免疫组化方法对２１虺先驱工斑狼疮患者皮损及非皮损皮肤中角朊细胞内的雌激素受体（ＥＲ）进行检测，同时取肿瘤等疾病患者的正常皮肤进行对照研究，结果均阴性，并对此结果进行了初步分析。     

Epidermal Langerhans cells,dermal dendritic cells,and keratinocytes in viral lesions of skin and mucous membranes: an immunohistochemical study

Summary We wanted to evaluate the eventual expression of viral antigens and MHC class II products by keratinocytes as well as the alterations of epidermal Langerhans cells and dermal dendritic cells in viral lesions of skin and mucous membranes. Therefore we investigated 68 biopsy specimens of protracted viral lesions, such as warts, condylomas, and mollusca contagiosa, and of rapidly resolving viral lesions such as herpes simplex virus infection. For this we used immunohistochemical staining techniques and several monoclonal and polyclonal antisera. In most cases investigated viral antigens (human papilloma virus antigens or herpes simplex virus type 1 antigens) could be demonstrated in keratinocytic nuclei. Except for a few viral lesions in which epidermal Langerhans cells were rather numerous, epidermal Langerhans cells were reduced in number or absent in almost all viral lesions. Moreover, epidermal Langerhans cells and dermal dendritic cells showed changes in morphology, distribution, and immunophenotype. These alterations may be caused by a toxic effect of the virus on dendritic cells. HLA-DR⁺ keratinocytes could be identified in few viral lesions onlyl HLA-DQ⁺ keratinocytes were not seen. Possible explanations for this lack of MHC class II expression by keratinocytes are discussed.Aspirant of the NFWO (National Fund for Scientific Research)     

Thrombomodulin expression on dermal cells in normal and psoriatic skin

Abstract The various subsets of dermal cells with a dendritic appearance can be identified by phenotypic differences in cell markers. We report on the morphology and tissue distribution of dermal cells detected with a monoclonal antibody against thrombomodulin in histological sections of normal arm and scalp skin and psoriatic skin. Double staining with antibodies to factor XIIIa, CD34 and CD68 was also employed in scalp biopsies to elucidate the relationship between thrombomodulin⁺ dermal cells and dermal dendrocytes and macrophages described by others. Thrombomodulin⁺ dermal cells in normal arm skin had little cytoplasm with fine branched dendrites and tended to be localized just beneath the epidermis. In scalp skin these cells had longer, more numerous dendrites and were distributed in the papillae and perivascular adventitial dermis primarily in the upper and central reticular dermis. In psoriatic skin, thrombomodulin⁺ dermal cells had an increased cytoplasmic volume with stout, less branched dendrites and appeared in the papillae and among inflammatory cells. Dermal cells detectable by thrombomodulin expression were factor XIIIa^–, CD34^– and CD68^–, and seemed to represent a distinct subset of dermal cells which may function in tissue repair. However, thrombomodulin⁺ dermal cells and factor XIIIa⁺ dendrocytes were frequently seen close together and could act cooperatively to regulate extravascular thrombin homeostasis in both normal and pathological dermal environments. Received: 26 May 1997     

MiRNA-203在寻常性银屑病皮损的表达及其对HaCaT细胞增殖的影响

目的 研究微小核糖核酸(miRNA)-203在寻常性银屑病患者皮损中的表达,并探讨对角质形成细胞株(HaCaT细胞)增殖的影响.方法 取2014-2016年23例寻常性银屑病患者的皮损组织和相邻非皮损组织.荧光定量PCR法检测组织中miRNA-203的表达水平,并以5'端、3'端地高辛标记的探针对皮肤组织切片中目的miRNA进行原位杂交,观察miRNA-203在皮肤组织中的定位情况.将miRNA-203模拟物(miRNA-203模拟物组)和miRNA-203模拟物阴性对照(阴性对照组)分别转染HaCaT细胞,正常细胞培养组作为空白对照组,采用噻唑蓝(MTT)法、流式细胞仪和Western印迹法分别对HaCaT细胞的增殖、细胞周期及相关周期蛋白(Cyclin D1、Cyclin B1)的变化进行检测.结果 miRNA-203特异性地表达在表皮的角质形成细胞中,除细胞核外,细胞质亦有表达,且寻常性银屑病患者皮损组织中miRNA-203表达水平(1.35±0.28)显著高于非皮损组织(0.52±0.09),差异有统计学意义(t=6.76,P=0.012).转染miRNA-203模拟物能抑制HaCaT细胞增殖(F=9.36,P=0.007),且空白对照组、阴性对照组和miRNA-203模拟物组HaCaT细胞增殖率均随时间的延长逐渐增加(F=18.68,P＜0.001).与阴性对照组和空白对照组相比,miRNA-203模拟物组HaCaT细胞被阻滞在G2/M期(G2/M期细胞比例:31.33％±4.56％比17.02％±3.53％、16.67％±3.32％,均P＜0.05),HaCaT细胞周期蛋白周期蛋白D1表达水平较高(1.15±0.13比0.52±0.05、0.56±0.07,均P＜0.05),而周期蛋白B1水平较低(0.43±0.08比0.93±0.16、0.91±0.0.15,均P＜0.05).结论 miRNA-203可能参与了寻常性银屑病的发生发展过程.     

The effects of epidermal keratinocytes and dermal fibroblasts on the formation of cutaneous basement membrane in three-dimensional culture systems

The cutaneous basement membrane (BM) plays an important role in normal and pathological conditions. However, few studies have addressed the formation of the cutaneous BM using three-dimensional culture systems. In this study, to elucidate the effects of human epidermal keratinocytes and dermal fibroblasts on the formation of the cutaneous BM, keratinocytes were cultured on several dermal substrates in the presence or absence of fibroblasts at the air–liquid interface. After 2 weeks of culture, immunohistochemical stainings for the components of the BM and electron microscopic studies of the BM zone (BMZ) were performed. In cultures of keratinocytes alone on dead reticular dermis or collagen gel without fibroblasts, 4 integrin chain, laminin, type IV and VII collagens were all expressed. However, ultrastructurally, BMZ was not formed. In cultures of keratinocytes on fibroblast-populated collagen matrix, laminin, and type IV and VII collagens were expressed more strongly than in the absence of fibroblasts. In addition, elements of the BMZ such as hemidesmosomes, lamina lucida, lamina densa and anchoring fibrils were formed, although it was still incomplete. In the culture of keratinocytes alone on de-epidermized dermis (DED) (surface up), 4 integrin chain, laminin, and type IV and VII collagens were strongly expressed. Also, the BMZ appeared similar to that in normal skin. In cocultures of keratinocytes and fibroblasts on DED or cultures of keratinocytes on DED combined with fibroblast-populated collagen matrix, type IV collagen was expressed more strongly than in cultures of keratinocytes alone. Ultrastructurally, similar findings to those of cultures of keratinocytes alone on DED were seen. Interestingly, when keratinocytes and fibroblasts were cocultured on DED, some fibroblasts were seen in the upper dermis as a result of migration into the dermis through partial loss of the lamina densa. These results show that keratinocytes produce most of the components of the BM such as laminin, and type IV and VII collagens. In addition, fibroblasts stimulate the expression of the components of the BM and the formation of a BMZ, suggesting that fibroblasts may produce laminin, and type IV and VII collagens or influence the effects of keratinocytes on the formation of the BM through a keratinocyte–fibroblast interaction.This investigation was supported by a grant (04-2001-027) from the Seoul National University Hospital Research Fund and partly by the Pacific Corporation.     

狼疮性肾炎患者肾组织FaS和Bax抗原的表达

为探讨Fas,Bax抗原的表达与狼疮性肾炎（lupus nephritis,LN)发病过程之间的关系，用免疫组化方法检测狼疮肾活检标本石蜡切片中Fas,Bax抗原的表达情况，狼疮性肾炎Bax抗原阳性表达率为70％，肾小管，肾小球，间质三个部位均有阳性表达，Fas抗原阳性表达率为75％，阳性表达仅见于肾小管，Fas,Bax阳性表达与LN病理分型间无明显的相关性（P＞0．05），促凋亡基因的过度表达，可能是狼疮性肾炎的发生，发展的重要因素。     

Fc gamma-receptors and HLA-DR antigens on endothelial cells in psoriatic skin lesions

Receptors for the Fc-part of IgG (FcR) and HLA-DR antigens on endothelial cells in normal and lesional skin from patients with psoriasis were studied in cryostat sections, using soluble immune complexes and monoclonal antibodies. FcR and HLA-DR antigens were detected on endothelial cells of dermal vessels both in sections of normal and lesional skin. The expression of FcR varied from one vessel to another and on endothelial cells within one and the same vessel. The expression of FcR and HLA-DR antigens was enhanced in sections of lesional skin compared with normal skin and most pronounced in lesional skin from active psoriasis. The enhanced expression may be mediated by interferon produced in psoriatic lesions. The presence of FcR and HLA-DR antigens on endothelial cells adds further evidence of he involvement of these cells in immune processes in the skin.