HPV16、18-DNA亚型检测早期诊断宫颈癌84例分析

目的:探讨HPV16、18-DNA亚型检测对宫颈癌的早期诊断价值。方法:84例标本分别采用FQ-PCR法和巴氏染色法。结果:HPV16、18-DNA阳性19例,阳性率为22.6%,19例HPV16、18-DNA阳性分泌物中4例为CIN2或以上。HPV16、18亚型是导致80%宫颈上皮瘤样病变的主要因素。讨论:HPV16、18-DNA检测可作为宫颈癌早期诊断的有效手段,能准确地预测宫颈癌发生可能性。     

通用引物PCR-SSCP技术分析16Sr RNA基因特征快速鉴定细菌

目的探讨通用引物聚合酶链反应-单链构象多态性(PCR-SSCP)技术在细菌检测中的应用.方法选取9种常见的细菌,采用通用引物对细菌的16SrRNA基因进行PCR扩增,产物进行单链构象多态性分析,并以标准菌株为对照.结果一对引物扩增产物SSCP图谱各细菌之间难以区别;两对引物扩增产物SSCP图谱条带数目、相对迁移率及条带之间的间距存在明显,可相互区别;3种细菌的标准菌株和临床菌株的SSCP图谱完全一致.结论采用针对16SrRNA的通用引物PCR-SSCP技术以标准菌株为对照可方便快捷地检测细菌.     

In vivo quantification of fluorescent molecular markers in real-time: A review to evaluate the performance of five existing methods

With the advent of molecular-targeted fluorescent markers, there is a renewed interest in fluorescence quantification methods that are based on continuous wave excitation and multi-spectral image acquisition. However, little is known about their in vivo quantification performance. We reviewed the performance of five selected methods by analytically describing these and varying input parameters of irradiance, excitation geometry, collection efficiency, autofluorescence, melanin content, blood volume, blood oxygenation and tissue scattering using optical properties representing those for human skin. We identified one method that corrects for variations in all parameters. This requires image acquisition before and after marker administration, under identical geometry. Hence, it is suited for applications where the site of interest can be relocated (e.g. anaesthetized animals and dermatology). For applications where relocation is not possible, we identified a second method where the uncertainty in the fluorescence signal was ±20%. Hence, use of these methods can substantially aid in vivo fluorescence quantification compared to use of the raw fluorescence signal, as this changed by more than 3 orders of magnitude. Since these methods can be computed in real-time, they are of particular interest for applications where direct feedback is critical, as diagnostic screening or image-guided surgery.     

胰腺癌中p16基因甲基化改变及其蛋白表达分析

目的:探讨胰腺癌与癌旁组织中p16基因启动子区异常甲基化的改变及其蛋白表达的特点,以及其与胰腺癌发生发展的关系。方法:分别采用免疫组化SP法及甲基化特异PCR(MSP)检测46例人胰腺癌和癌旁组织中p16基因表达及其甲基化的水平,并结合临床资料进行分析。结果:胰腺癌中p16蛋白表达率为41.3%(19/46),而癌旁组织表达率为95.7%(44/46),两者有差异显著性(P<0.01)。p16蛋白阳性的19例胰腺癌标本中均未检出基因甲基化;p16蛋白缺失的27例标本中有18例检出基因甲基化,甲基化率为39.1%。p16基因甲基化与蛋白缺失有明显关系(P<0.05)。p16基因表达及其启动子区甲基化的发生率与胰腺癌的大小,患者性别﹑年龄的差异无显著意义(P>0.05),但与组织分化程度﹑淋巴结转移﹑PTNM分期有显著意义(P<0.05)。结论:p16基因启动子的异常甲基化可影响p16蛋白的表达,它们与胰腺癌的发生发展有关;p16甲基化和蛋白表达可能成为胰腺癌诊断及预后的候选标志物之一。     

P53、P16、Bcl-2基因及产物在原发性肺癌中的表达及其临床病理意义

目的 :探讨原发性肺癌 Mtp5 3、p16、Bcl- 2的异常表达与肺癌发生、发展的关系 ,以及它们之间的调控关系。 方法 :用免疫组化技术检测 (L SAB) 114例原发性肺癌组织中 p5 3、p16、Bcl- 2的表达。并用 PCR技术对 6 2例 p16蛋白丢失的肺癌组织中 p16外显子 2 (p16 E2 )的缺失进行分析。 结果:p5 3蛋白异常表达与肺癌组织学类型、分化程度无关 (P >0 .0 5 ) ,但与淋巴结转移情况有关 (P >0 .0 5 )。 p16蛋白阳性表达率与肺癌的组织学类型无关 ,但是其表达水平与非小细胞肺癌 (NSCL C)的细胞分化程度和淋巴结转移密切相关 (P <0 .0 5 )。 NSCL C中 p16E2的缺失率为 45 .2 %,其缺失水平随淋巴结转移和组织学分级的升高而升高 (P <0 .0 5 )。 SCL C(小细胞肺癌 )和正常肺组织中无 p16 E2的缺失。 Bcl- 2在小细胞肺癌中阳性率 6 2 .2 %显著高于 Sq Ca(鳞癌 )的 2 5 %和 Ad Ca(腺癌 )的 9.1%(P <0 .0 5 ) ,与细胞分化程度和淋巴结转移情况无关。另外 ,p5 3与 p16蛋白之间存在调控关系。 结论 :(1) p16、p5 3、Bcl- 2的异常参与肺癌的发生、发展过程。 (2 ) p16基因及产物的异常表达与 NSCL C的组织学分级和淋巴结转移相关 ,可能参与 NSCL C的发生、发展和转移过程。 (3) Bcl- 2反映 SCL C的分之生物学行为和临床     

LightCycler实时监测PCR定量分析血清HBV DNA

目的 检验LightCycler实时监测PCR（real-time detection PCR,RTD-PCR)对血清中HBV DNA定量检测的灵敏性和可重复性，探讨HBV血清标志物与HBV DNA定量的关系。方法 HBV定量按深圳匹基公司乙肝PCR荧光检测试剂盒使用说明，对乙肝标志物已明确的773例血清中HBV DNA定量结果进行统计分析。结果 实时监测PCR对血清中HBV DNA定量检测的灵敏性高，可检测低至1000拷贝／ml血清；可重复性好，批间误差＜20％；各种标志物类型的血清HBV DNA含量分布情况表明，HBV血清标志物中HBeAg与HBV DNA含量有明显的关系，一般HBeAg阳性血清HBV DNA含量较高，但也有相当一部分例外。结论 LightCycler实时监测PCR对血清中HBV DNA定量检测灵敏性高可重复性好；仅根据HBV血清标志物往往不能确定乙肝患者HBV DNA复制水平的高低，HBV DNA定量检测具有十分重要的临床意义。     

弥漫性大B细胞淋巴瘤的病理、免疫组化特征及IgH基因重排的检测

目的 探讨弥漫性大B细胞淋巴瘤(DLBCL)临床和病理组织学特征以及免疫组化特异性抗体及IgH基因重排检测在其诊断和鉴别诊断中的价值。方法 收集30例弥漫性大B细胞淋巴瘤及其临床资料，用免疫组化孓P法标记LCA，CD20，CD79a，CD30及bcl-2抗体和用PCR方法检测15例IgH基因重排。结果 70％(21／30)弥漫性大B细胞淋巴瘤发病年龄在40～70岁，淋巴结内外都可累及。组织病理学：中心母细胞淋巴瘤占83．3％(25／30)，免疫母细胞淋巴瘤占3．3％(1／30)，间变性大细胞淋巴瘤占6．7％(2／30)，及富于T细胞B细胞淋巴瘤占6．7％(2／30)。免疫标记LCA均表现阳性，CD20、CD79a、CD30、bcl-2表达率分别为86．7％(26／30)，93．3％(28／30)，6．7％(2／30)，20％(6／30)。15例DLBCL中IgH基因重排阳性为66．7％(10／15)。结论 弥漫性大B细胞淋巴瘤是一组异质性肿瘤，必须结合其组织病理学形态和特异抗体的免疫组化检测，对一些疑难病例需做IgH基因重排检测以进行诊断和鉴别诊断。     

The role of HLA‐DQβ1 alleles in susceptibility to rheumatoid arthritis

Aim: Rheumatoid arthritis (RA) is the most common chronic inflammatory erosive joint disease with the worldwide distribution of approximately 0.5–1.0%. Etiology of RA is not exactly known but immunologic and genetic factors play an important role in the pathogenesis of the disease. Genetic factors such as human leukocyte antigens (HLA) are responsible for many autoimmune diseases; therefore we decided to look for a correlation between RA and the presence of HLA‐DQβ1 alleles as possible genetic markers. Methods: Genomic DNA from the whole blood samples of 25 patients with RA and 86 normal individuals as control group were extracted by salting out method. The genomic DNA was amplified by polymerase chain reaction‐sequence specific primer (PCR‐SSP) technique. HLA‐typing was done by this method after optimizing the PCR reaction for each allele. In this procedure seven serological subclasses of HLA‐DQβ1 can be detected. Results: Comparing the results between the patients and controls show a significant increase in the frequency of HLA‐DQ8 (*0302, *0305) alleles in RA patients. The P‐values were 0.007 and the relative risk for these alleles was evaluated higher than 1. Conclusions: The results suggest that DQ8 is the dominant HLA‐DQβ1 allele that is associated with susceptibility to RA in north‐eastern Iran.