胆脂瘤上皮过度增殖行为的免疫组化研究

目的　探讨中耳胆脂瘤上皮的过度增殖性和生长方式。方法　用免疫组化SP法观察了表皮生长因子受体 (epidermalgrowthfactorreceptor,EGFR) ,增殖细胞核抗原Ki6 7、Ⅳ型胶原蛋白(collagenIV)和层粘连蛋白 (laminin ,LN)在 18例胆脂瘤标本的表达 ,并与 8例外耳道正常皮肤相比较。结果　EGFR有两种染色图案 ,棕黄色的线状或颗粒状胞膜强染色和颗粒状的胞浆染色 ,胆脂瘤标本染色均强于皮肤。在Ki6 7的免疫染色中 ,阳性细胞核为棕黄色 ,胆脂瘤上皮的阳性细胞核比皮肤多。CollagenIV和LN图案极其相似 ,为连续的棕黄色线状和 (或 )带状图案。 4例胆脂瘤上皮基膜下结缔组织中的血管比皮肤丰富。结论　EGFR和Ki6 7在胆脂瘤上皮的表达从不同角度反映了胆脂瘤上皮的过度增殖性。CollagenIV和LN的表达说明胆脂瘤属良性病变 ,血液供应可能比皮肤丰富     

Localization of laminin isoforms in the guinea pig cochlea

OBJECTIVE: To elucidate the pathogenesis of inner ear disorders by examining the distribution of a major component of basement membranes, the glycoprotein laminin. STUDY DESIGN: Animal model. METHODS: Adult guinea pig cochleae were fixed with methanol and sectioned. Sections were examined for the presence of laminins alpha2, alpha5, beta1, and gamma1 using immunohistochemistry. RESULTS: Laminins alpha2, alpha5, and beta1 were not detected. Laminin gamma1 was found in the limbus, spiral ligament, and stria vascularis. CONCLUSIONS: Anti-laminin antibodies have been found in patients with inner ear disorders. Laminin gamma1 demonstrates specific staining at multiple sites in the guinea pig cochlea. These structures may be targets of antibodies causing sensorineural hearing loss.     

GFAP,FVIII/RAg,Laminin, and fibronectin in gliosarcomas: An immunohistochemical study

Summary GFAP, Factor VIII/RAg, laminin, and fibronectin were immunohistochemically investigated in 15 glioblastomas and 15 gliosarcomas. GFAP was found variably positive in the glial areas. F VIII/RAg characterizes the endothelial cells and in gliosarcomas suggests the origin of the sarcomatous component from the endothelial proliferations. Laminin separates the two components and characterizes the inner and the outer basement membranes in the vessels. It is multiplied and thickened in endothelial proliferations, while it is often fragmented in the larger vessel wall proliferations. Our observations confirm that gliosarcoma represents the last stage of a process which starts with the endothelial hyperplasia of glioblastoma.Partially supported by CNR, Special Project Center, Grant no. 82.00410.96     

层粘素和纤粘素在胃癌中的分布

采用层粘素（ＬＮ）多抗和纤粘素（ＦＮ）多抗检测７６例胃癌。证实癌细胞分化程度越低和局部浸润能力越强，造成的基底膜（ＢＭ）缺损越严重。证明反应性间质增生与癌肿的浸润转移趋势有关。癌细胞穿透基底膜进入间质，引起间质反应性增生，后者可促进癌细胞的浸润和转移。     

Viability and induction of tyrosine aminotransferase in rainbow trout hepatocytes cultured on laminin and polylysine in a serum-free medium

The objective of this study was to find an economical substrate for long-term primary culture of rainbow trout hepatocytes in which viability and differentiated liver function could be maintained in a serum-free media. Attachment efficiency of primary cultures of rainbow trout (Oncorhynchus mykiss) hepatocytes was determined for cells grown on plastic and Falcon Primaria dishes, and dishes coated with ECM (extracellular matrix), Matrigel, fibronectin, collagen type 1, laminin and polylysine. On ECM, Matrigel, laminin and polylysine, attachment efficiency exceeded 90%. On plastic, Falcon Primaria, fibronectin and collagen-coated dishes attachment efficiency was 15–20% and the cells were easily dislodged by pipetting. Cells grown on ECM, laminin and polylysine were maintained up to 22 day with 76%, 60% and 80% viability, respectively. On ECM cells flattened and formed a confluent monolayer on day 2 in culture. Cells grown on matrigel, laminin and polylysine had a more rounded appearance with establishment of cell-to-cell contacts on the second day in culture. Induction of TAT (tyrosine aminotransferase), a marker enzyme for differentiated liver cell functions, was studied with DEX (dexamethasone) in cells grown on laminin and polylysine in serum-free media. Cells grown on laminin and polylysine showed TAT inducibility up to 15 and 19 days, respectively.     

Rapid repair and regeneration of damaged rabbit sciatic nerves by tissue-engineered scaffold made from nano-silver and collagen type I

A tissue-engineered scaffold with nano-silver and collagen type I was constructed and investigated for its ability to adsorb laminin and the usefulness in the repair and regeneration of damaged peripheral nerves in animals. The nano-silver scaffold displayed ideal microtubule structure under electronic microscope; even distribution of the nano-silver particles was also seen with energy spectrometry. After immersion in a laminin solution, the laminin-attached scaffolds were implanted into rabbits to repair a 10-mm injury of the sciatic nerve. At 30 days post-implantation, regeneration of the damaged nerve was evaluated by transmission electron microscopy, electrophysiological examination and fluoro-gold (FG) retrograde labelling. Compared with the control collagen-scaffold without nano-silver, the nano-silver-containing scaffold showed a higher rate of laminin adsorption, regenerated a nerve with a thicker myelin sheath and improved the nerve conduction velocity and nerve potential amplitude. FG retrograde labelled the newly grown axons in the spinal cord cortex anterior horn and the dorsal root ganglion. These results demonstrate the superior functionality of the nano-silver-collagen scaffold in the adsorption to laminin and subsequent regeneration of damaged peripheral nerves.     

Immunohistochemical study of merosin-negative congenital muscular dystrophy: laminin α2 deficiency in skin biopsy

We studied the immunohistochemical expression of laminin subunits α2, α1, β1 in muscle and skin biopsy samples from three patients with congenital muscular dystrophy (CMD), and from ten control patients investigated for various neuromuscular disorders. Merosin α2 chain was not detectable in the basement membrane of muscle fibers, or in the nerve endings, cutaneous nerves, and corium in the skin of the CMD patients, whereas it was clearly expressed in the skin biopsy samples from control patients, especially in the nerve endings of the arrector pili muscles. Laminin α1 chain was expressed in the corium, in the muscle fiber membranes of arrector pili muscles and in cutaneous nerve fibers, perineurium and blood vessels in controls and in CMD patients. Laminin β1 chain was faintly expressed in the corium, and a diffuse labeling was detected on arrector pili muscle with enhanced expression at nerve endings, intracutaneous nerves and capillaries, with similar findings in all biopsy specimens. For merosin-negative CMD patients, skin biopsy may provide a diagnostic alternative to muscle biopsy since merosin deficiency can be demonstrated in the skin neural structures, and in particular in the nerve endings of the arrector pili smooth muscles. Received: 29 October 1996 / Revised, accepted: 27 January 1997     

自体微粒皮与异体脱细胞微粒真皮混合移植对大鼠创面纤维连接蛋白和层粘连蛋白表达的影响

目的 探讨自体微粒皮与异体脱细胞微粒真皮混合移植对创面愈合的影响,并对有关机制做进一步研究.方法 Wistar大鼠作为供体,SD大鼠为受体,在SD大鼠背部建立全层皮肤损伤模型.90只SD大鼠分为5组,每组18只,第1组为自体微粒皮组;第2组为异体脱细胞微粒真皮移植组;第3、4、5组为混合移植组.混合移植组中自异体微粒皮的面积比例分别为:1∶1、1∶0.5、1∶0.25.术后第2、3、4周分别测量每组创面的愈合率,采集创面标本,做HE染色,检测纤维连接蛋白(FN)和层粘连蛋白(LN)、进行组间比较.结果 混合移植组与自体微粒皮移植组比较,混合移植组创面愈合率及FN、LN均高于自体微粒皮组,其中1∶0.25混合移植组最高,差异有统计学意义(P＜0.05或P＜0.01).结论 混合移植创面愈合率高于自体微粒皮移植,且自体微粒皮与异体脱细胞微粒真皮混合移植的面积比例按1∶0.25效果最佳,这可能与创面纤维连接蛋白和层粘连蛋白升高有关.     

复方壳多糖组织工程皮肤的组织学特性

目的评价制备的复方壳多糖组织工程皮肤在组织学方面的特性，为其修复皮肤缺损创面提供实验依据。方法通过苏木精-伊红（HE）染色、高碘酸-希夫（PAS）染色、免疫组织化学染色及电镜，观察组织工程皮肤的组织学特征，包括表皮、真皮及两者的连接结构基底膜。结果制备的复方壳多糖组织工程皮肤表皮细胞增殖活跃，分层分化良好，厚约150μm，真皮层细胞生长正常，排列有序。PAS染色示真表皮间有均匀红染的条带出现，免疫组织化学染色结果显示Ⅳ型胶原、Ⅶ型胶原、层黏蛋白反应阳性，在电镜下可见角质形成细胞间大量桥粒，基底层内侧较多半桥粒，与机体皮肤结构相似。结论复方壳多糖组织工程皮肤组织结构良好，符合新型皮肤替代物在治疗皮肤缺损时的组织学要求。     

血清HA、PLD、PCⅢ、LN、Ⅳ—C联合检测对慢性肝病的诊断价值探讨

目的:寻找肝纤维化等慢性肝病敏感可靠的血清学诊断指标。方法:对196例各型肝病联合检测血清透明质酸(HA)、脯氨酸肽酶(PLD)、Ⅲ型前胶原(PCⅢ)、层粘蛋白(LN)、Ⅳ型胶原(Ⅳ-C),并与10例肝组织学和30例健康献血员做对照研究。结果:五项指标分别在急性肝炎、慢性肝炎和肝硬变中依次升高,差异显著(P<0.05或P<0.01),其改变情况与轻、中、重型纤维化程度呈同步性,有良好的正相关。结论:五项指标联合检测可极大地增加慢性肝病临床诊断的准确性与可靠性,动态观察其变化可掌握病情演变。