P^16蛋白,增殖细胞核抗原在肾癌的表达及与预后的关系

目的　探讨多肿瘤抑制基因Ｐ１６／ＭＴＳ１蛋白在肾细胞癌（ＲＣＣ）的表达及与临床病理指标、预后的关系。方法　采用免疫组化方法对Ｐ１６蛋白和增殖细胞核抗原（ＰＣＮＡ）在５６例ＲＣＣ的表达进行病理观察。结果　Ｐ１６蛋白在ＲＣＣ的阳性率为６０．７％（３４／５６）,各组织学分级的阳性率差异有显著性（Ｐ＜０．０１）,其阳性组和阴性组的５年生存率差异无显著性。ＰＣＮＡ标记指数（ＰＣＮＡＬＩ）在１％～８５％,平均为１９．７％,在不同组织学分级及临床分期中差异有显著性（Ｐ＜０．０１）。ＰＣＮＡ高表达组和ＰＣＮＡ低表达组的５年生存率差异有显著性（Ｐ＜０．０５）。Ｐ１６蛋白的表达与ＰＣＮＡ无一致性（Ｐ＜０．０５）。结论　ＰＣＮＡＬＩ是反映ＲＣＣ组织分化程度及预后的良好指标,Ｐ１６蛋白的胞浆型表达对肾癌临床分期及预后无明显影响。     

PCR和分离培养检测细胞培养支原体污染的比较

目的：检测细胞培养中支原体的污染,建立支原体通用的ＰＣＲ方法。方法：根据已出版的序列设计并合成一对支原体（柔膜体纲）１６ＳｒＲＮＡ基因组特异引物。对实验室所存的１２种支原体、大肠杆菌、变形杆菌及无支原体污染的组织培养细胞进行了ＰＣＲ扩增。以ＰＣＲ与分离培养比较检测了３３份细胞培养标本。结果：１２种支原体均出现了约６２０ｂｐ左右的特异性扩增带,敏感性为１００～１０００个支原体细胞,且常见的６种细胞培养污染支原体的扩增片段均能被ＨｉｎｄⅢ切成２８０与３４０ｂｐ左右的２条带。理论上推测这对引物可扩增所有支原体１６ＳｒＤＮＡ。但对大肠杆菌、变形杆菌及无支原体污染的组织培养细胞等未见有扩增带出现。在对３３份细胞培养标本的检测中,除ＰＣＲ和分离培养同时检出２份阳性外,前方法还另检出阳性３份。５份ＰＣＲ扩增物均被ＨｉｎｄⅢ切出２条带。结论：ＰＣＲ较分离培养敏感性高,可提高细胞培养污染支原体的检出率,防止出现假阴性结果;如做好质量控制,对于一般实验室检测细胞培养支原体污染,ＰＣＲ扩增较分离培养具有更大的应用意义     

聚合酶链反应检测细菌16S rRNA基因

根据细菌１６ＳｒＲＮＡ基因的高度保守性,设计合成所有细菌、革兰氏阳性细菌及革兰氏阴性细菌的共同引物,采用聚合酶链反应检测已知细菌１３株,三对引物分别扩增的阳性率为１００％,倍比稀释法能检出细菌的最低浓度为４ＣＦＵ·ｍｌ－１,同时检测临床样本４０份,阳性率为６７５％（２７／４０）,同期细菌培养阳性率为４５％（１８／４０）,二者比较差异有显著性（Ｐ＜０．０５）。结果提示聚合酶链反应检测细菌１６ＳｒＲＮＡ基因具有高度的特异性和敏感性。     

Ginkgo biloba extract EGb761 reduces the development of amphetamine-induced behavioral sensitization: effects on hippocampal type II corticosteroid receptors

Pretreatment of rats with the extract of Ginkgo biloba termed EGb761 reduced the behavioral sensitization induced by successive -amphetamine administrations (0.5 mg/kg) as estimated by increasing values of locomotor activity. EGb761 pretreatment also prevented the reduced density of [³H]dexamethasone binding sites in the dentate gyrus and the CA1 hippocampal regions of -amphetamine treated animals. These observations suggest that EGb761, by reducing glucocorticoid levels, could modulate the activity of the neuronal systems involved in the expression of the behavioral sensitization.     

子宫平滑肌肿瘤P16抑癌基因表达的定量研究与临床病理关系

目的 :为了探明抑癌基因 P16在子宫平滑肌瘤发生发展中的作用及其在诊断中的临床意义。方法 :应用细胞免疫荧光技术及流式细胞术 ,对 5 5例子宫平滑肌肿瘤石蜡组织块的 P16表达定量分析。结果 :正常子宫平滑肌组织 P16表达均为阴性。 14例平滑肌瘤仅 2例出现过度表达 (14 % ) ;交界性平滑肌肿瘤 12例 ;多数为过度表达 (75 % ) ;平滑肌肉瘤 2 9例中 ,则有 2 7例 (93 % )出现过度表达。从平滑肌瘤、交界性平滑肌瘤到平滑肌肉瘤 ,P16表达量显著递增。P16表达量随肿瘤细胞分裂相、异形性、密度的增高也有明显增高。P16低表达者 5年生存率为 85 % ,而过度表达者为 4 8.5 %。结论 :子宫平滑肌肿瘤一旦出现 P16过度表达应考虑为恶性 ;P16表达可能和平滑肌瘤向平滑肌肉瘤的转化密切相关 ,P16表达可作为判断病人预后的一项客观指标。     

Human papillomavirus type 16 DMA in an odontogenic keratocyst

DNA was extracted from an odontogenic keratocyst and assayed for the presence of human papilloma virus (HPV) type 16 DNA sequences using high stringency Southern blot hybridization. HPV type 16 homologous DNA sequences were detected at a copy number of 50-100 genome copies per diploid cell. The oral HPV DNA was not identical to the prototype HPV 16 when cleaved with the restriction enzyme Pst-I, since it appeared to lack the Pst-I C fragment (L2/L1 ORFs) and contained "off-sized" high molecular weight fragments suggestive of integration events into the host cell chromosome.     

p16和p15第二外显子HapⅡ位点在成人急性髓性白血病中的突变

目的　探讨p16和p15基因第二外显子HapⅡ位点在成人急性髓性白血病 (AML)中的突变。方法　用甲基敏感和甲基非敏感限制性内切酶切消化基因组DNA后 ,经PCR扩增和琼脂糖电泳研究 3 1例AML中无p16/p15基因第二外显子缺失的患者中HapⅡ位点的突变情况。结果　在 3 1例成人AML样本中 ,p16E2 的 4个HapⅡ位点和p15E2 的 6个HapⅡ位点均未发生突变。结论　本研究提示p16和p15基因第二外显子中因甲基化而引起的CCGG位点的突变在成人AML患者中不是主要失活方式。     

Epithelial mucin expression in bladder cancer: correlation with pathological and clinical parameters

Recently, attention has been drawn to the role of polymorphic epithelial mucin (PEM) as a possible target for cancer immunotherapy. To investigate the expression of this molecule in bladder tissue, we used two mouse monoclonal antibodies (HMFG1 and HMFG2) raised against the core protein of the PEM. The localization of these two anti-PEM antibodies was examined in normal (n=10), inflammatory (n=10) and malignant (n=67) bladder tissue samples with the use of a three-step avidin-biotin method. For HMFG1 and HMFG2 localization was successful in 78% and 60% of the bladder cancer samples, respectively, where as they were localized only in 30% and 40% of normal bladder tissue samples, respectively. Staining of either antibodies did not correlate with the grade, stage, or survival of bladder cancer patients. We conclude that PEM is frequently overexpressed by bladder cancer cells and HMFG1 is the antibody of choice to be used as a carrier of a cytotoxic agent for application of intravesical targeted therapy of bladder cancer. Received: 25 August 1999 / Accepted: 1 March 2000     

子宫内膜癌中p16、E-cad mRNA表达的研究

目的 :探讨p16及E 钙粘附蛋白 (E cad)基因mRNA在子宫内膜癌中的表达及意义。方法 :采用RT PCR技术检测 3 0例子宫内膜癌及 8例正常子宫内膜组织的p16及E cadmRNA的表达。结果 :①子宫内膜癌p16mRNA的阳性表达明显低于正常子宫内膜 (P <0 .0 1) ,但p16mRNA的表达与子宫内膜癌的组织病理分级无关 ;②E cadmRNA在子宫内膜癌中呈低表达 ,正常组织内呈强阳性表达 ,二者差异非常显著 (P <0 .0 1) ,并且在子宫内膜癌中E cadmRNA的阳性表达与组织病理学分级呈负相关 (P <0 .0 5 )。结论 :p16、E cadmRNA的表达在子宫内膜癌的发生及进展过程中起重要作用