血尿细胞角质蛋白20的表达

目的探讨血尿患者尿脱落细胞细胞角质蛋白20（CK20）的表达及其意义。方法采用逆转录-聚合酶链反应（RT-PCR）对47例首发症状为血尿的尿路移行细胞癌患者、18例以血尿就诊的非泌尿系肿瘤患者以及10例健康志愿者尿脱落细胞CK20的表达进行检测。结果47例尿路移行细胞癌中有38例脱落细胞CK20阳性，对照组无CK20阳性表达。结论检测尿脱落细胞CK20表达，对诊断尿路移行细胞癌敏感性和特异性高，可作为血尿患者筛选尿路移行细胞癌的方法之一。     

P504S联合CK34βE12、p63检测在前列腺癌诊断中的意义

目的：评价P504S、CK34βE12、p63检测在前列腺癌诊断中的应用价值。方法：采用（EliVisionTM-plus）免疫组化方法,观察P504S、CK34βE12、p63在前列腺癌、前列腺增生中的表达情况。结果：CK34βE12和p63在良性前列腺增生中阳性率100%,少数腺泡周围间断表达,个别腺腔阴性表达;前列腺癌仅1例腺泡周围间断表达（P〈0.01）。P504S在良性前列腺增生中阳性率17.6%,为弱阳性;在前列腺癌中阳性率100%,多为强阳性（P〈0.01）。结论：P504S联合CK34βE12、p63检测可提高前列腺癌诊断的准确率。     

紫外线灭活的EB病毒蛋白对抗角蛋白自身抗体产生的调节作用

目的研究紫外线(UV)灭活和热处理的EB病毒刺激培养的人脐带血B细胞产生抗角蛋白自身抗体(AKautoAb)的作用。方法用淋巴细胞分离液分离人脐带血单一核细胞,L-亮氨酸甲酯去除单核细胞、NK细胞和细胞毒性T细胞,2-氨乙基硫脲溴化物处理的绵羊红细胞去除T细胞,从而获得纯化B细胞。用UV灭活和热处理EBV处理培养的B细胞。流式细胞仪检测UV灭活EBV组细胞CD5、CD3、CD4和CD8的表达。ELISA法检测培养上清液中AKautoAbIgG和IgM的产生,并与EBV转化B细胞产生的AKautoAbIgG和IgM作对比。结果细胞培养14d时,未检测到T细胞,CD5 B细胞占43%;28d时,CD5 B细胞占47%。UV灭活EBV组AKautoAbIgG18d以后各时间点有显著变化(P<0.05),AKautoAbIgM26d与其他时间点差异有显著性(P<0.05)。热处理EBV组AKautoAbIgG和IgM无明显变化(P>0.05)。EBV转化B细胞40d,AKautoAbIgG和IgM的产生与同期的UV灭活EBV组差异有显著性(P<0.05)。结论UV灭活EBV有诱导AKautoAb产生的作用,而热处理EBV不能诱导抗体的产生,提示灭活EBV诱导AKautoAb产生可能是通过EBV的蛋白成分实现的。     

角蛋白16和17在部分表皮肿瘤中的表达和意义

目的 研究角蛋白16 和17（K16、K17）在部分表皮肿瘤中的表达和意义。方法 采用免疫组化法检测K16、K17在正常皮肤、脂溢性角化病、日光性角化病、皮角、基底细胞癌、鳞状细胞癌中的表达。结果 K16、K17 在正常皮肤表皮各层呈阴性表达;在脂溢性角化病表皮各层中除了角质层外大多呈阳性表达;在日光性角化病表皮K16 均不表达,K17 除了角质层外在其他各层也不表达;K16 在皮角表皮棘层呈强阳性表达,但在其他各层均呈阴性表达,K17 在皮角表皮各层基本呈阳性或强阳性表达;在基底细胞癌中,K16、K17除了在棘层呈弱阳性表达外,在其他各层均呈阴性表达;在鳞状细胞癌中,K16 在基底层呈阴性表达,但在其他各层及K17 在表皮全层均呈阳性或强阳性表达。K16 除基底层外在肿瘤表皮其他各层表达比较差异均有统计学意义（均P＜0.05）;K17 在表皮各层表达差异均有统计学意义（均P＜0.05）。结论 K16、K17的表达状况可为表皮肿瘤的鉴别诊断提供病理学方面的依据。     

瘢痕表皮角蛋白的变化规律及正常表皮细胞培养液对成纤维细胞胶原合成的影响

目的：研究增生性瘢痕和正常皮肤表皮角蛋白的变化规律以及正常表皮细胞培养液对瘢痕和正常皮肤成纤维细胞增殖和胶原合成的影响。方法：4例烧伤患者,每人取少量增生性瘢痕和临近的正常皮肤组织,用免疫组化法观察两种组织中与细胞增殖有关的角蛋白14、与细胞分化有关的角蛋白10和与细胞活化有关角蛋白16的变化。收集正常表皮细胞培养12h后的无血清培养液,用DMEM依次配成体积分数为0、5％、10％和20％,分别加入到加有正常和瘢痕组织成纤维细胞的培养板中。分别以MTT法测定吸光度（A）值,Beckman液闪计数仪测定3H-脯氨酸掺人值,分别用以反映成纤维细胞的增殖和胶原合成速率。结果：与正常表皮比较,瘢痕表皮中角蛋白16明显表达增强,角蛋白10降低,角蛋白14无明显变化;正常表皮细胞培养液能促进正常皮肤和瘢痕成纤维细胞的增殖并抑制胶原的合成,但对两种成纤维细胞的作用程度不尽相同,对正常成纤维细胞增殖的促进作用和对胶原合成的抑制作用都较瘢痕成纤维细胞显著。结论：瘢痕表皮的活化可能是瘢痕增生的一个重要因素;正常表皮细胞培养液既能促进成纤维细胞增殖,还能抑制胶原的合成,但对瘢痕成纤维细胞胶原合成的抑制作用较正常成纤维细胞弱。     

Elevated Serum Levels of Cell Death Circulating Biomarkers,M30 and M65, In Patients with β-Thalassemia Major

Deposition of iron in visceral organs, mainly in the liver, causes tissue damage in β-thalassemia major (β-TM) patients. Keratin 18 (K18) represents one of the major caspase substrates during apoptosis of hepatocytes. To better characterize the hepatic apoptosis and/or necrosis in β-thal patients, the circulating levels of M65 (soluble intact K18) and M30 (the caspases-generated K18 fragment) were measured in 40 β-TM patients and compared with 40 healthy controls. The ratio of M30/M65 (caspase-cleaved to total K18) was also determined in thalassemic and normal subjects. Results of the ELISA assays revealed that the serum levels of hepatocyte death markers, M65 and M30, were significantly increased in β-thal patients compared to healthy controls (p <0.0001). M30 serum levels were also positively correlated with the serum levels of liver transaminases including aspartate aminotransferase (AST) (r = 0.337, p = 0.047) and alanine aminotransferase (ALT) (r =0.391, p = 0.02).     

Role of phosphorylation in ethanol-induced aggregation of keratin intermediate filaments

BACKGROUND: Keratins are members of a diverse group of tissue-specific cytoskeletal components known as intermediate filaments. Regulation of the structure and intracellular distribution of intermediate filaments is known to be related to the phosphorylation state of their structural subunits. It also is known that disruption of the keratin filaments of hepatocytes in response to chronic ethanol ingestion is characteristic of alcoholic liver disease. METHODS: To characterize the mechanism of ethanol-induced keratin filament reorganization and dephosphorylation, cells were grown in culture with and without ethanol, and then were treated at the end of the incubation period for 1 hr with either 8-bromo-adenosine 3':5'-cyclic monophosphate (8Br), water-soluble forskolin (ws-forskolin), H-89 diHCL, or okadaic acid. Morphology of the cells was examined by immunofluorescence microscopy, and keratin phosphorylation levels were determined by analysis of 32p labeling. RESULTS: We found that treatment of hepatoma cells with 300 mM ethanol results in disruption and aggregation of the keratin network in the vicinity of the nucleus as well as a hypophosphorylation of keratin subunits from ethanol-treated cells compared with non-ethanol-treated controls. 8Br and ws-forskolin treatment of ethanol groups restored keratin phosphorylation to control levels and reversed the ethanol-induced aggregation of keratin filaments. When H-89, an inhibitor of A-kinase, was added to control cells, keratin filament disorganization and dephosphorylation was observed. H-89 produced only a slight additional decrease in keratin phosphorylation in ethanol-treated cells, with no change in keratin distribution. Okadaic acid treatment of control cells produced hyperphosphorylation and filament network disruption, whereas in ethanol groups a reversal of the ethanol-mediated hypophosphorylation was observed but without reversal of the keratin filament aggregation. CONCLUSIONS: These results suggest that site-specific phosphorylation of keratin filaments is important in maintaining their integrity and that activation of the A-kinase system can antagonize the effects of ethanol, whereas its inhibition results in filament dephosphorylation and reorganization, mimicking effects of ethanol treatment.     

细胞角蛋白CK34βE12对子宫内膜良恶性病变的鉴别作用研究

目的研究细胞角蛋白CK34βE12在子宫内膜良恶性病变中的表达情况，探讨鉴别诊断子宫内膜良恶性病变的方法。方法选择子宫内膜病变蜡块标本158例，应用SP免疫组织化学染色法进行染色观察，根据病变性质分组进行统计学分析。结果CK34βE12阳性表达率在子宫内膜普通性增生性病变组为90．36％（75／83），其中，单纯性增生组为93．02％（40／43）、腺囊性增生组为87．50％（35／40）、非典型增生组为76．74％（33／43）；子宫内膜腺癌组为9．38％（3／32）。CK34βE12阳性表达率普通性增生性病变组高于子宫内膜腺癌组，非典型增生组高于子宫内膜腺癌组，差异均有统计学意义（P〈0．001）。普通性增生性病变组CK34βE12阳性表达率高于非典型增生组，但差异无统计学意义（／9〉0．05）。结论CK34βE12检测可作为鉴别诊断子宫内膜良恶性病变的辅助方法。