尖锐湿疣凝集素标记免疫组化研究

我们应用ＣｏｎＡ、双花扁豆凝集素（ＤＢＡ）、ＬＣＡ、蓖麻凝集素（ＲＣＡ １２０）、ＳＢＡ、ＰＮＡ、ＵＥＡ １和ＷＧＡ等８种凝集素为细胞膜结合糖分子探针，采用免疫组织化学ＡＢＣ法，观察了尖锐湿疣表皮细胞膜凝集素结合特点，以探讨其变化的意义和在诊断中的应...     

红斑狼疮皮损中浸润细胞免疫表型和角质形成细胞HLA-DR抗原表达的研究

应用抗ＨＬＡＤＲ、ＣＤ３、ＣＤ４、ＣＤ８、ＣＤ２０的单克隆抗体和ｓｔｒｅｐｔｒａｖｉｄｉｎｐｅｒｏｘｉｄａｓｅｓｔａｉｎｉｎｇ（ＳＰ）技术对１０名正常人皮肤,１６例ＳＬＥ皮损和１９例ＤＬＥ皮损进行了免疫组化研究。观察到正常人皮肤角质形成细胞未见ＨＬＡＤＲ抗原表达,而ＳＬＥ（６／１６）,ＤＬＥ（８／１９）皮损处角质形成细胞可以表达ＨＬＡＤＲ抗原。在ＳＬＥ、ＤＬＥ真皮内浸润细胞主要为Ｔ淋巴细胞（ＣＤ３＋浸润细胞）,且以ＴＨ细胞（ＣＤ４＋浸润细胞）占优势。另外,还发现在两种ＬＥ表皮角质形成细胞表达ＨＬＡＤＲ抗原处,真皮内可见ＣＤ３＋浸润细胞和激活的Ｔ淋巴细胞（ＨＬＡＤＲ＋浸润细胞）。讨论了ＬＥ皮损角质形成细胞ＨＬＡＤＲ抗原表达及其与病损内浸润细胞免疫表型的关系。ＬＥ皮损处ＨＬＡＤＲ＋角质形成细胞可能具有抗原递呈作用,而角质形成细胞异常表达ＨＬＡＤＲ抗原则可能与真皮内浸润单个核细胞或淋巴细胞释放的ＩＦＮα,ＴＮＦγ等有关。     

ABC免疫光镜及免疫电镜证实硬红斑皮损炎症区郎格罕细胞

应用Ｌｅｕ６、ＨＬＡ－ＤＲ、Ｓ１００抗体ＡＢＣ免疫光镜及免疫电镜方法对ＥＩ皮损内Ｓ１００蛋白阳性组织细胞性质进行了研究。８例ＥＩ免疫光镜观察显示ＥＩ真皮深部及皮下炎症区Ｓ１００蛋白阳性组织细胞其Ｌｅｕ６及ＨＬＡ－ＤＲ染色亦阳性，且多呈树枝状或椭圆形。３例ＥＩ免疫电镜观察显示Ｌｅｕ６十细胞其细胞核是卷曲的脑回状，胞浆内可见不典型的棒状样的郎格罕颗粒，部分阳性细胞超微结构有受损伤的表现。上述结果表明，ＥＩ皮损内所谓的Ｓ１００蛋白阳性组织细胞为ＬＣ，ＬＣ不仅存在于表皮及真皮，亦存在于皮下组织中，且可能介导了ＥＩ细胞免疫的发生。     

系统性红斑狼疮患者外周血单一核细胞CD23的表达

为了揭示Ｂ细胞中ＣＤ２３的表达与ＳＬＥ发生发展的关系及在ＳＬＥ发病机理中可能的作用，我们应用ＡＢＣ免疫组化法和斑点核酸杂交技术对ＳＬＥ患者外周血单一核细胞（ＰＢＭＣ）ＣＤ２３蛋白和ｍＲＮＡ表达进行了检测。结果显示：３０例ＳＬＥ患者ＰＢＭＣＣＤ２３蛋白表达显著增高（Ｐ＜０．０１），且与疾病活动呈正相关关系（ｒｓ＝０．３８１４，Ｐ＜０．０５）；具有不同ＡＮＡ、抗ｄｓＤＮＡ抗体水平，有无伴肾损、脑损的ＳＬＥ患者，ＰＢＭＣＣＤ２３表达均无显著性差异（Ｐ均＞０．０５）；单纯使用皮质类固醇激素治疗或和其它免疫抑制剂联合治疗的ＳＬＥ患者，ＰＢＭＣＣＤ２３表达亦无显著性差异（Ｐ＞０．０５）。２０例ＳＬＥ患者ＰＢＭＣＣＤ２３ｍＲＮＡ表达较正常人显著增高（Ｐ＜０．０１）。经治疗病情稳定后，ＣＤ２３蛋白和ｍＲＮＡ表达均降至正常（Ｐ均＞０．０５）。提示在ＳＬＥ活动期Ｂ细胞高度激活、增殖并大量表达ＣＤ２３，且该种表达与ＡＮＡ、抗ｄｓＤＮＡ抗体产生水平无直接关系     

系统性红斑狼疮患者红细胞参数的分析

应用ＣｏｌｔｅｒＪＴ－ＩＲ型细胞分析仪对４０例红斑狼疮患者（ＳＬＥ）的红细胞相关参数进行分析。结果显示：与对照组比较ＳＬＥ患者均存在不同程度的变化。稳定期与活动期比较红细胞计数（ＲＢＣ），血红蛋白（ＨＧＢ）、平均红细胞体积（ＭＣＶ）、红细胞积（ＨＣＴ）、红细胞体积分布宽度（ＲＤＷ）之间存在显著性差异（Ｐ〈０．０１）。同时对且红细胞免疫功能进行检测，实验结果表明，ＳＬＥ患者红细胞免疫功能低下。并对Ｓ     

系统性红斑狼疮患者外周血单一核细胞中Th1型细胞因子的表达

目的 探讨系统性红斑狼疮（ＳＬＥ）患者中Ｔｈ１型细胞因子表达的情况。方法 用逆转录聚合酶链反应（ＲＴ－ＰＣＲ）法检测了ＳＬＥ患者外周血单一核细胞（ＰＢＭＣ）中白介素２（ＩＬ－２）ｍＲＮＡ表达（４１例）和γ干扰素（ＩＦＮ－γ）ｍＲＮＡ表达（３１例）的水平。结果 与正常人相比,活动期ＳＬＥ患者中ＩＬ－２表达水平降低者占７８．０％,ＩＦＮ－γ表达水平降低者占８３．９％。活动期ＳＬＥ组ＰＢＭＣ中ＩＬ－２和     

系统性红斑狼疮患者外周血淋巴细胞bcl-2的表达及临床意义

目的 为了探讨ｂｃｌ－２基因在系统红斑狼疮（ＳＬＥ）发病机制中的作用及临床意义。方法 应用逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）检测３１例ＳＬＥ患者外周血单一核细胞（ＰＢＭＣ）ｂｃｌ－２ｍＲＮＡ表达水平和流式细胞仪双标记法分析其Ｔ、Ｂ细胞ｂｃｌ－２蛋白表达。结果 活动期ＳＬＥ患者ＰＢＭＣｂｃｌ－２ｍＲＮＡ表达水平明显升高,占５５．６％,且活动期ＳＬＥ患者ＣＤ３＾＋、ＣＤ４＾＋和ＣＤ８＾＋Ｔ细胞亚群Ｐ     

SLE患者外周血单个核细胞中IL-6 IL-10表达研究

目的 探讨系统性红斑狼疮（ＳＬＥ）患者中Ｔｈ２型细胞因子ｍＲＮＡ的表达。方法 用逆转录－多聚酶链反应（ＲＴ－ＰＣＲ）法检测了１０例正常人和１５例活动期ＳＬＥ患者外周血单个核细胞（ＰＢＭＣ）中白介素６（ＩＬ－６）和白介素１０（ＩＬ－１０）ｍＲＮＡ表达的水平。结果 与正常相比,活动期ＳＬＥ患者ＰＢＭＣ中ＩＬ－６和ＩＬ－１０ｍＲＮＡ的表达水平均增高（均Ｐ〈０．０１）。结论 ＳＬＥ患者中Ｔｈ２型细胞因子Ｉ     

SLE患者外周血单个核细胞中IL—10与γ干扰素的表达

为了探讨系统性红斑狼疮（ＳＬＥ）患者中ＴＨ２／ＴＨ１型细胞因子自然表达的趋势,用逆转录聚合酶链反应（ＲＴ－ＰＣＲ）法检测了１０名正常人和１５例活动期ＳＬＥ患外周血单个核细胞（ＰＢＭＣ）中白介素１０（ＩＬ－１０）和γ干扰素（ＩＦＮγ）ｍＲＮＡ表达的水平。结果,与正常人相比,活动期ＳＬＥ患者ＰＢＭＣ中ＩＬ－１０ｍＲＮＡ的表达水平增高,而ＩＦＮγｍＲＮＡ的表达水平降低（均Ｐ〈０．０１）,表明活动性ＳＬＥ     

成人皮肌炎伴钙盐沉着1例

患男，３２岁，干部。１９９０年４月８日，因面部发红，四肢无力，行走困难诊于某医科大学。实验室检查：ＷＢＣ１６×１０９／Ｌ、Ｈｂ１１０、ＥＳＲ３２、ＣＰＫ（Ｒｏｓａｌｋｉ法）９０ｕ／Ｌ、ＡＬＤ１６０８ｎｍｌＳ－１／Ｌ、ＬＤＨ１７１ｕｍｏｌＳ－１／...     

皮脂腺肿瘤凝集素亲合组化研究

本研究采用１７种生物素化的凝集素，应用ＡＢＣ法研究了２３例正常皮肤和２２例皮脂腺良、恶性肿瘤的肿瘤细胞凝集素受体的定位及分布，发现ＬＣＡ在皮脂腺癌的阳性率较高；正常皮脂腺、皮脂腺痣、皮脂腺增生、皮脂腺腺瘤可见胞膜及胞浆的海绵、网状着染；皮脂腺癌中胞浆失去这种规则着染，着染形状及分布极不规则，说明在皮脂腺恶性转化过程中，细胞表面复合糖糖基及细胞的正常结构发生了明显的变化。     

Comparison of proliferating cells between human adult and fetal eccrine sweat glands

Studies of sweat glands had demonstrated that there were degenerating cells and proliferating cells in the eccrine sweat glands. To compare the differences in the proliferating cells between human adult and fetal eccrine sweat glands, immunostaining of proliferating-associated proliferating cell nuclear antigen (PCNA) and Ki67 nuclear antigen (Ki67) was performed, and the location and the percentage of the positive staining cells were analyzed. The results showed that a few cells of the secretory and ductal portion in both the adult and fetal eccrine sweat glands stained positive with Ki67 and PCNA. The labeling index of PCNA in adult eccrine sweat glands was 34.71 ± 8.37%, while that in the fetal was 62.72 ± 6.54%. The labeling index of PCNA in fetal eccrine sweat glands was higher than that in adult. Myoepithelial cells were negative staining with anti-PCNA antibody in adult eccrine sweat glands, while in the fetal a few myoepithelial cells were positive staining. Labeling index of Ki67 in adult eccrine sweat glands was similar to that in the fetal, ranging from 0.5 to 4.3%. Myoepithelial cells of the adult and fetal eccrine sweat glands both were negative staining with anti-Ki67 antibody. We concluded that the myoepithelial cells had proliferating ability only in fetal eccrine sweat glands, and that the proliferating ability of fetal eccrine sweat glands was stronger than that of the adult. Competing interest statement: The authors declare that they have no competing financial interests.     

Differential expression of collagen integrin receptor on fetal vs. adult skin fibroblasts: implication in wound contraction during healing

BACKGROUND: Fetal skin wound healing is characterized by an absence of contraction and scar formation, two important observations associated with adult healing often leading to pathological problems. OBJECTIVES: We have studied the capacity of adult and fetal human skin fibroblasts to contract collagen gels, collagen being the major structural component of dermal matrix. METHODS: In parallel with collagen gel contraction studies, we have used fluorescence-activated cell sorter analysis to study the levels of collagen receptors expressed at the surface of fibroblasts derived from fetal or adult skin samples. RESULTS: Strong differences were detected between freshly isolated fetal and adult fibroblasts. Fetal fibroblasts had a very low capacity to contract collagen gel, whereas adult cells significantly contracted gels in the same conditions. The expression of alpha1, alpha2 and alpha3 integrin subunits was also significantly different depending of the donor age: alpha1 and alpha3 integrin subunit expression was lower in fetal cells compared with adult cells, whereas alpha2 integrin subunit expression was higher. When grown in monolayers, adult cells showed rapid changes in their contractile capacity and integrin expression while fetal cells were only affected after several passages. CONCLUSIONS: These observations indicate that intrinsic differences between fetal and adult fibroblasts can strongly influence the quality of wound repair.     

Immunolocalization of epidermal growth factor receptors in normal developing human skin.

The embryogenesis of normal human skin is a complex process involving multiple cell types and developmentally regulated growth factors. The immunohistochemical localization of epidermal growth factor receptors (EGF-R) was studied in human fetal skin because this receptor modulates all known actions of EGF and TGF-alpha. EGF-R are present in developing skin as early as the 42nd day of gestation. Immunoreactive EGF-R are present in keratinocytes, endothelial, and skeletal muscle cells. In contrast to normal adult human skin in which the EGF-R are primarily restricted to the basal and immediately suprabasal keratinocytes, the fetal epidermis showed a persistent expression of EGF-R in all cell layers. The absence of EGF-R on the outer, apical surface of periderm cells that are exposed to amniotic fluid was unexpected and may reflect down-regulation of EGF-R by EGF/TGF-alpha or related fetal growth factors present in amniotic fluid. The complex regulation of EGF-R in embryonic hair follicles and sebaceous glands indicates an active and perhaps regulatory role for EGF/TGF-alpha in the development and function of pilosebaceous glands as well as mammalian skin in general.     

Distribution of metallothionein in normal and pathological human skin

The expression and distribution of metallothionein (MT) in frozen sections of normal and pathological human skin was studied using the monoclonal antibody L2E3 directed against MT derived from human fetal liver. Immunohistochemical staining of normal fetal and adult skin revealed strong reactivity in basal keratinocytes of epidermis and outer hair root sheath, hair matrix cells and the secretory coil, but not the exocrine portion of eccrine glands; myoepithelial cells around apocrine sweat glands were similarly stained. In epidermal hyperplasia, variable numbers of suprabasal keratinocytes were stained, whereas in interface dermatitis, interrupted staining was found in the basal layer. Weak or scattered staining was observed in squamous tumours, whereas basal cell carcinomas did not show consistent staining. The distribution of MT in normal skin was in line with the germinative role of basal keratinocytes and hair matrix cells, whereas its distribution in hyperplastic epidermis was in line with experimental animal data, and reflected the increase in the germinative pool in these conditions. It is concluded that monoclonal antibody L2E3 may serve as a valuable immunohistochemical marker in diagnostic cutaneous pathology since it labels basal keratinocytes selectively, and since it discriminates between eccrine and apocrine sweat glands.     

Immunohistochemical studies on epithelial cells in mixed tumor of the skin

We performed studies on the lectin-binding pattern in epithelial tumor cell components of 4 cases of mixed tumor of the skin developing on the face in addition to identification of keratin and carcinoembryonic antigen (CEA), compared with those of normal sweat glands. Normal eccrine glands showed specific labelling with Dolichos biflorus agglutinin (DBA) and soybean agglutinin (SBA), whereas none of the studied lectins reacted specifically with normal apocrine glands. In mixed tumors the dark cells, which form the inner layer of the tubuloalveolar and ductal structure, showed the presence of keratin and CEA, as well as of specific sugar structures that bind to DBA and SBA. On the other hand the light cells that form the outer layer of the tubular structures or the solid epithelial cell nests gave only a faint to moderate staining of keratin, and no staining of CEA or lectins. It is probable that the inner dark cells differentiate toward the cells that have the same sugar structures on the cell surface as those of the normal eccrine gland cells, while the outer light cells appear to be immature or in a less differentiated state.     

不同剂量UVA1辐射硬皮病鼠模型的研究

目的 探讨不同剂量的UVA1辐射硬皮病鼠模型后皮肤厚度和胶原含量的变化。方法 用100 μL（400 μg/mL）的博来霉素皮下注射BALB/c小鼠背部,共4周,建立硬皮病的小鼠模型,然后将小鼠随机分为空白对照组、模型对照组、UVA1照射组（100 J/cm2、60 J/cm2、20 J/cm2）及阴性对照组。每周照射3次,共10周,观察并比较各组小鼠皮肤组织病理改变、皮肤厚度和胶原含量的变化。结果 模型对照组与空白对照组相比,皮肤厚度（t = 4.945,P < 0.001）和胶原含量（t = 3.712,P < 0.01）的差异均有统计学意义。UVA1照射组小鼠皮肤变软、变薄,但只有HD-UVA1组小鼠皮肤厚度、胶原含量与模型对照组、阴性对照组比较,差异有统计学意义（P < 0.05）。照射组之间相互比较,皮肤厚度差异F = 14.853,P < 0.01,胶原含量差异F = 6.317,P < 0.01,尤其是HD-UVA1与MD-UVA1和LD-UVA1之间有显著差异。结论 HD-UVA1照射能明显改善博来霉素所致的硬皮病鼠模型的皮肤厚度、胶原含量的变化,可能与其抑制皮肤的胶原增生、降低胶原含量有关。     

Akt和mTOR在皮肤基底细胞癌中的表达

目的 检测Akt和雷帕霉素靶蛋白（mammalian target of rapamycin, mTOR）在皮肤基底细胞癌( basal cell carcinoma, BCC)中的表达情况,探讨Akt/mTOR信号通路在BCC发病机制中的作用及其临床意义。方法 免疫组织化学方法检测37例BCC以及16例正常皮肤标本中的Akt和mTOR蛋白的表达情况。结果 BCC中Akt蛋白的阳性表达率为62.2%,mTOR蛋白的阳性表达率为83.8%,与正常皮肤组织中Akt蛋白的阳性表达率12.5%和mTOR蛋白的阳性表达率37.5%相比,显著增高(均为P<0.05)。Akt和mTOR的表达水平在BCC中存在正相关,有统计学意义(均为P<0.05)。结论 Akt和mTOR在BCC中的过度表达,提示Akt/mTOR通路可能是BCC发生发展的机制之一。     

Ultrastructural localization of lectin-binding sites in normal human eccrine and apocrine glands

The distribution of carbohydrate residues in eccrine and apocrine glands of normal human skin was studied using a post-embedding technique with Lowicryl K4M. Thin sections were incubated with Ulex europaeus agglutinin I (UEA I), wheat germ agglutinin (WGA), peanut agglutinin (PNA), concanavalin A (Con A), soybean agglutinin (SBA), and dolichos biflorus agglutinin (DBA). All lectins except for PNA showed labeling of the plasma membranes of dark cells, clear cells, and apocrine cells. The granules of the eccrine gland were labeled with all lectins except for DBA. The mitochondrial granules of the apocrine gland were not labeled with any lectin, whereas the lysosomal granules showed a positive reaction with all lectins except for PNA. After incubation with PNA, in eccrine glands the granules were the only structure labeled, whereas in apocrine glands the luminal side of the plasma membrane and cytoplasmic vesicles beneath it were the only structures labeled.     

Changes of expression of intermediate filament proteins during ontogenesis of eccrine sweat glands.

The intermediate filament expression in fetal and adult human eccrine sweat glands was studied by immunoperoxidase microscopy performed on cryostat sections using monoclonal antibodies against various cytokeratins (CK), vimentin, and actin. In palmar skin of 14-week-old fetuses, the early dermal cords showed a primitive CK pattern similar to that of epidermal basal cells. From week 15 on (distal finger skin), inner cells of the proximal (ductal) portion of the glandular anlagen expressed CK 1/10/11 and 19 (markers of adult eccrine ductal luminal cells). In addition, CK 4 was expressed in ductal luminal cells mainly in the fetal period. In the distal portion of the sweat gland anlagen the increased or new expression of the simple-epithelium-type CK 7, 8, 18, and 19 was detected at week 15, indicating the onset of the secretory differentiation pathway. Two subsegments of the prospective secretory portion could be distinguished (elongated part and end bud). Interestingly, in fetuses, most secretory portion cells co-expressed vimentin in addition to CK. From week 22 on, peripheral cells of the secretory portion were stained for CK 17 and smooth-muscle-type actin, suggesting myoepithelial differentiation. In newborn and adult eccrine glands, secretory cells expressed mainly CK 7, 8, 18, and 19, whereas myoepithelial cells were conspicuous by their co-expression of certain CK (including CK 5 and 17), vimentin, and smooth-muscle-type actin and sometimes even glial filament protein (GFP), similar to myoepithelial cells of other glands. These results throw further light onto the complex processes of fetal development of eccrine sweat glands and their cellular diversification. The possible biologic significance of the differential CK expression in the various glandular cell types is discussed.