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陕西医学杂志 2010年第8期39卷 950-953页

作者：安小亮王瑞陕西中医学院附属医院麻醉科咸阳712000 西安市中心医院麻醉科

目的:研究大鼠全脑缺血再灌注后增殖细胞核抗原PCNA的表达时程及其与凋亡的关系。方法:采用4-VO法建立全脑缺血再灌注损伤模型,将各组各时间点脑组织切片分别进行HE染色:光镜下观察海马回CA1区神经元形态结构改变;免疫组化染色:观察PCN... 详细信息

目的:研究大鼠全脑缺血再灌注后增殖细胞核抗原PCNA的表达时程及其与凋亡的关系。方法:采用4-VO法建立全脑缺血再灌注损伤模型,将各组各时间点脑组织切片分别进行HE染色:光镜下观察海马回CA1区神经元形态结构改变;免疫组化染色:观察PCNA蛋白在CA1区的表达情况;TUNEL染色:观察海马回CA1区神经细胞凋亡情况,统计阳性细胞表达情况,计算细胞凋亡指数;将PCNA蛋白免疫组化切片进行图象分析测得平均灰度值。结果:PCNA蛋白于再灌注后2h在海马CA1区可见表达下降,再灌注后6～24h组明显下降,之后持续低表达。缺血再灌注后6h凋亡细胞开始增加,主要位于CA1区,24～48h为高峰,至72h明显减少。结论:PCNA蛋白主要在缺血再灌注后早期即24h前DNA损伤未积累到严重程度时执行抗凋亡的作用,对损伤神经元进行修复。

关键词：再灌注损伤/免疫学增殖细胞核抗原/免疫学模型,动物细胞凋亡脑大鼠

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增殖核抗原在胆管上皮细胞的表达

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临床小儿外科杂志 2009年第2期8卷 27-28页

作者：陈风汪健黄顺根王兴东上海南汇医院普外科 201300 苏州大学附属儿童医院普外科浙江省215003

目的通过先天性胆总管囊肿，胆管癌和流产胎儿胆管标本的免疫组织化学染色，探讨增殖核抗原（PCNA）在胆管黏膜上皮细胞的表达，了解胆管上皮细胞的增殖分化情况。方法取先天性胆总管囊肿标本20例，胆管癌标本8例，流产胎儿胆管标本20... 详细信息

目的通过先天性胆总管囊肿，胆管癌和流产胎儿胆管标本的免疫组织化学染色，探讨增殖核抗原（PCNA）在胆管黏膜上皮细胞的表达，了解胆管上皮细胞的增殖分化情况。方法取先天性胆总管囊肿标本20例，胆管癌标本8例，流产胎儿胆管标本20例，用福尔马林固定24h。标本常规石蜡包埋，切片，切片厚度3μm。行PCNA免疫组织化学染色。结果PCNA阳性染色时细胞核为棕色。正常胎儿胆管的表达为（5．45±1．90）％，先天性胆总管囊肿的表达为（47．75±5．48）％，胆管癌的表达为（83．88±7．24）％。3组之间比较，差异有统计学意义。结论PCNA在3种细胞的表达不同，先天性胆总管囊肿的上皮细胞处于增殖状态。

关键词：增殖细胞核抗原/免疫学胆管肿瘤/病因学胆总管囊肿/病因学胆管

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Survivin基因和增殖细胞核抗原在大肠癌组织中表达及其意义

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陕西医学杂志 2007年第11期36卷 1478-1480页

作者：许静洪成延萍魏晓丽刘敏丽张生军魏茂富延安大学医学院延安716000

目的:探讨Survivin和增殖细胞核抗原PCNA在大肠癌组织中的表达和意义。方法:采用免疫组织化学方法检测正常大肠粘膜、腺瘤和腺癌组织中Survivin和PCNA表达。结果;Survivin在正常大肠粘膜无表达,在大肠腺癌和腺瘤组织中表达的阳性率分别... 详细信息

目的:探讨Survivin和增殖细胞核抗原PCNA在大肠癌组织中的表达和意义。方法:采用免疫组织化学方法检测正常大肠粘膜、腺瘤和腺癌组织中Survivin和PCNA表达。结果;Survivin在正常大肠粘膜无表达,在大肠腺癌和腺瘤组织中表达的阳性率分别为70.9%和62.5%;大肠癌组和大肠腺瘤组OD值显著高于正常大肠粘膜组;大肠癌组中PCNA-LI为(50.2±13.2)%,显著高于正常大肠粘膜组(12.6±3.1)%和大肠腺瘤组(33.2±7.9)%;大肠癌组中Survivin阳性的PCNA-LI为(61.3±9.3)%,显著高于Survivin阴性的PCNA-LI(36.4±6.6)%。结论:Survivin和PCNA的高表达在大肠癌的发生、发展中起重要作用,PCNA可反映大肠癌的增殖状况,两者可作为判断其预后的参考指标。

关键词：肠肿瘤/免疫学 @Survivin基因增殖细胞核抗原/免疫学细胞凋亡免疫组织化学

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Cancer and autoimmunity: autoimmune and rheumatic features in patients with malignancies

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ANNALS OF THE RHEUMATIC DISEASES 2001年第5期60卷 433-440页

作者： Abu-Shakra, M Buskila, D Ehrenfeld, M Conrad, K Shoenfeld, Y Soroka Med Ctr Dept Med IL-84101 Beer Sheva Israel Soroka Med Ctr Rheumat Dis Unit IL-84101 Beer Sheva Israel Ben Gurion Univ Negev IL-84105 Beer Sheva Israel Chaim Sheba Med Ctr Res Ctr Autoimmune Dis IL-52621 Tel Hashomer Israel Tel Aviv Univ Sackler Fac Med IL-69978 Tel Aviv Israel Tech Univ Dresden Inst Immunol D-8027 Dresden Germany

Objectives-To review the autoimmune and rheumatic manifestations of patients with malignancy. Methods-A Medline search of all published papers using keywords related to malignancies, autoimmunity, rheumatic diseases, and paraneoplastic syndromes. Results-Patients with malignant diseases may develop autoimmune phenomena ana rheumatic diseases as a result of (a) generation of autoantibodies against various autoantigens, including oncoproteins (P185, 1-myc, c-myc, c-myb), tumour suppression genes (P53), proliferation associated antigens (cyclin A, B1, D1, E;CENP-F;CDK, U3-RNP), onconeural antigens (Hu, Yo, Ri, Tr), cancer/testis antigens (IMAGE, GAGE, RAGE, SSX, ESO, SCP, CT7), and rheumatic disease associated antigens (RNP, Sm). The clinical significance of the various autoantibodies is not clear. Anti-oncoprotein and anti-tumour suppression gene antigens are detected before the diagnosis of the cancer or in the early stages of the malignant disease, suggesting a potential diagnostic or prognostic role. Anti-onconeural antibodies are pathogenic and are associated with specific clinical neurological syndromes (anti-Hu syndrome and others). (b) Paraneoplastic syndromes,a wide range of clinical syndromes, including classic autoimmune rheumatic diseases that develop among patients with cancer. (r) Rheumatism after chemotherapy, a clinical entity characterised by the development of musculoskeletal symptoms after combination chemotherapy for malignancy Conclusion-Autoimmune and rheumatic features are not rare among patients with malignancies. They are the result of various diverse mechanisms and occasionally they may be associated with serious clinical entities.

关键词：抗体抗核/免疫学抗原肿瘤/免疫学抗肿瘤联合化疗方案/副作用自身抗体/免疫学自身免疫疾病/病因学自身免疫疾病/免疫学血液肿瘤/药物疗法血液肿瘤/免疫学干扰素类/副作用肿瘤/药物疗法肿瘤/免疫学癌基因蛋白质类/免疫学类肿瘤综合征/药物疗法类肿瘤综合征/免疫学增殖细胞核抗原/免疫学风湿性疾病/病因学风湿性疾病/免疫学女(雌)性人类男(雄)性

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Immunoelectron microscopy of PCNA as an efficient marker for studying replication times and sites during pollen development

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CHROMOSOMA 2000年第6期109卷 397-409页

作者： González-Melendi, P Testillano, PS Ahmadian, P Reyes, J Risueño, MC CSIC Ctr Invest Biol Lab Nucl Org During Plant Dev E-28006 Madrid Spain Ctr Ingeneria Genet & Biotechnol La Habana Cuba John Innes Ctr Plant Sci Res Dept Cell Biol Norwich NR4 7UH Norfolk England

Here we report for the first time the ultrastructural localization of DNA replication sites in the nucleus of plant cells and the timing of replication through the pollen developmental programme by proliferating cell nuclear antigen (PCNA) immunogold labelling. Replication sites were identified by labelling with anti-PCNA antibodies in fibrils of the interchromatin region close to the condensed chromatin, defining a perichromatin sub-domain in the interchromatin space where DNA replication takes place. The same nuclear structures are decorated by anti-BrdU (5-bromo-2'-deoxyuridine) immunogold after short pulses of BrdU labelling. Double immunogold labelling for PCNA and DNA show colocalization on these perichromatin structures. PCNA immunoelectron microscopy also allows correlation of replicative activity with the dynamics of chromatin condensation. DNA replication was also monitored at different phases during pollen development by PCNA immunoelectron microscopy, revealing two peaks of DNA synthesis, at the beginning (early tetrad), and the end (late vacuolate), of microspore interphase. High-resolution autoradiography after [H-3]thymidine incorporation also showed high replicative activity at the same two periods of microspore interphase. In the bicellular pollen grain, PCNA immunogold labelling revealed that DNA replication in the generative cell starts at an intermediate stage of pollen maturation, whereas the vegetative nucleus does not replicate and is arrested in G1. The use of anti-PCNA antibodies at the ultrastructural level is an easier, faster and more feasible method than the detection of in vivo-incorporated nucleotides, especially in plant systems with long cell cycles. PCNA immunogold labelling is, there fore, proposed as an efficient marker for mapping the sites and timing of replication at the electron microscopy level.

关键词：放射自显影术/方法溴脱氧尿苷/代谢染色质/代谢染色质/超微结构 DNA复制/生理学免疫印迹法分裂间期显微镜检查免疫电子/方法有丝分裂洋葱/遗传学花粉/遗传学花粉/生长和发育增殖细胞核抗原/分析增殖细胞核抗原/免疫学增殖细胞核抗原/代谢胸苷/代谢

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3-methylcholanthrene triggers the differentiation of alveolar tumor cells from canine bronchial basal cells and an altered p53 gene promotes their clonal expansion

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CARCINOGENESIS 2000年第8期21卷 1477-1484页

作者： TenHave-Opbroek, AAW Shi, XB Gumerlock, PH Leiden Univ Med Ctr Dept Pulm NL-2300 RC Leiden Netherlands Univ Calif Davis Med Ctr Dept Internal Med Sacramento CA 95817 USA Univ Calif Davis Med Ctr Dept Urol Sacramento CA 95817 USA

Alveolar type II cells arising in canine bronchial autografts following exposure to 3-methylcholanthrene (MCA) give rise to carcinomas of varying glandular and squamous growth patterns. To study the role of the tumor suppressor gene p53 in this process, sections from progressive lesions were immunostained for p53 protein;microdissected regions were screened for p53 mutations. Adjacent sections were examined for type II cell expression [cuboid cell shape, large roundish nucleus, cytoplasmic staining for surfactant protein A (SP-A)] and proliferating cell nuclear antigen expression. Evidence for an altered p53 function (nuclear staining, missense mutations) was found in most carcinomas of all histologic types and in all grades of bronchial dysplasia, but not in hyperplastic or normal bronchial epithelium. It was primarily associated with the hyperplastic type II cell populations present in the basal zone of the lesions. In addition, we found SP-A staining in hyperplastic (but not in normal) bronchial basal cells. These data suggest that MCA initiates type II cell differentiation through phenotypic selection (basal cells). Inactivation of the p53 gene promotes the clonal expansion of the type II cells into discernible populations of (squamous or glandular) alveolar tumor cells. This in vivo study is the first to show that p53 is involved in a specific pathway leading to bronchogenic carcinoma.

关键词：支气管/病理学支气管/移植致癌物癌支气管原/化学诱导癌支气管原/遗传学癌支气管原/病理学细胞分化/药物作用细胞转化肿瘤/药物作用细胞转化肿瘤/遗传学细胞转化肿瘤/病理学克隆细胞基因 p53/遗传学肺肿瘤/化学诱导肺肿瘤/遗传学肺肿瘤/病理学甲基胆蒽小鼠裸突变误义点突变增殖细胞核抗原/分析增殖细胞核抗原/免疫学蛋白脂质类/分析蛋白脂质类/免疫学肺泡/药物作用肺泡/代谢肺泡/病理学肺表面活性物质相关蛋白质A 肺表面活性物质相关蛋白质类肺表面活性剂/分析肺表面活性剂/免疫学肿瘤抑制蛋白质p53/遗传学肿瘤抑制蛋白质p53/代谢动物狗小鼠

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Presentation of autoantibody to proliferating cell nuclear antigen in patients with chronic hepatitis B and C virus infection

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ANNALS OF THE RHEUMATIC DISEASES 1999年第10期58卷 630-634页

作者： Tzang, BS Chen, TY Hsu, TC Liu, YC Tsay, GJ Chung Shan Med & Dent Coll Dept Med Taichung 402 Taiwan Natl Tsing Hua Univ Dept Life Sci Hsinchu Taiwan

Objectives-To study the association of antibodies to proliferating cell nuclear antigen (PCNA) in patients with chronic hepatitis B (HBV) and C (HCV) virus infection. Methods-Sera from 243 patients with chronic HBV infection;379 patients with chronic HCV infection;80 patients with systemic lupus erythematosus (SLE);28 patients with rheumatoid arthritis;15 patients with Sjogren's syndrome;eight with polymyositis;eight with primary biliary cirrhosis;and 33 healthy control subjects were tested for the presentation of anti-PCNA antibodies by enzyme Linked immunosorbent assay (ELISA) and immunoblotting using recombinant PCNA as antigen. The distribution of immunoglobulin isotypes of anti-PCNA antibody was measured by ELISA assay. Results-By ELISA, anti-PCNA antibodies mere detected in 30 (12.3%) patients with chronic HBV infection, 71 (18.7%) patients with chronic HCV infection, and five (6.3%) patients with SLE. The inhibition of binding with these sera by purified PCNA was shown to exceed 71%. By immunoblotting, the frequency of anti-PCNA in patients with chronic HBV and HCV infection was 17 of 243 (7%) and 41 of 379 (11%), respectively. Absorption studies on indirect immunofluorescence showed the typical nuclear speckled staining pattern by anti-PCNA sera was abolished by preincubation of sera with PCNA. Anti-PCNA antibody was not: detected in sera from patients with autoimmune diseases except SLE. Anti-PCNA antibodies in patients with chronic HBV and HCV infection were predominantly IgG. Conclusion-These data suggest that anti-PCNA antibody are also present in patients with chronic HBV and HCV infection. Anti-PCNA antibody may not be specific for SLE.

关键词：关节炎类风湿/免疫学自身抗体/血液 CHO细胞病例对照研究仓鼠亚科酶联免疫吸附测定荧光抗体技术间接乙型肝炎慢性/免疫学丙型肝炎慢性/免疫学免疫印迹法免疫球蛋白G/血液肝硬化胆汁性/免疫学红斑狼疮系统性/免疫学多发性肌炎/免疫学增殖细胞核抗原/免疫学干燥综合征/免疫学动物人类

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