河北汉族人群D7S21位点DNA多态性研究

目的探讨D7S2 1位点在河北汉族人群分布的多态性 ,为DNA指纹数据库的构建及其法医学应用提供基础资料。方法应用MVR PCR方法和聚丙烯酰胺梯度凝胶电泳银染法对 12 4名河北汉族人群无关个体D7S2 1位点进行了快速检测 ,并进行数字编码。结果每一个体平均得到 3 6个数字编码 ,未发现任何两个无关个体所有编码相同 ,两无关个体 3 6个编码相同的机率为 3 .4 8× 10 -18。三种重复单位a 型、t 型和o 型出现的机率分别为 4 8.5 %、4 9.5 %和 2 .1%。该位点杂合度为 0 .9876,非父排除率为 0 .974 6,多态性信息含量为 0 .9872。结论D7S2 1位点在河北汉族人群中具有高度的多态性 ,聚丙烯酰胺梯度凝胶电泳银染法简便、快速 ,具有一定的实用价值     

Reevaluation of the Correlation between Angiotensinogen Gene M235T Polymorphism and Familial Essential Hypertension Using MS-PCR Technique

Renin- angiotensin system( RAS) candidat-ing gene,such as angiotensinogen( Ag T) as wellas angiotensin- converting enzyme( ACE) genepolymorphism were thought to have positive association with various cardiovasculardiseases.In1 992 ,Jeunemaitre[1] detected a microsatellitepolymorphism in the region of the an-giotensinogen gene which has linked to occur-rence of essential hypertension.Further analysisled to detection of several mutations in the cod-ing region of angiotensinogen gene[2 ] . Ma…     

云南个旧和宣威地区肺癌p53突变检测研究

目的　云南个旧及宣威地区是云南省肺癌的两大点状高发区 ,个旧以矿工为高发人群 ,宣威以农民为高发 ,该两地区肺癌发生的病因已初步阐明 ,本研究旨在深入探讨其发病的分子生物学基础。方法　对 2 6例个旧矿工肺癌患者 ,15例宣威农民肺癌患者 ,应用聚合酶链反应 -单链构象多态性法研究 p5 3第 5至第 8外显子突变 ,并对有突变的个体进行测序。结果　发现云锡矿工肺癌患者中有 13例、宣威农民肺癌患者中有 4例发生 p5 3突变 ,应用 DNA测序 ,发现云锡矿工肺癌中有 1例于第 135位密码子发生突变 ,由 GCC→ GTC,导致氨基酸改变 ,由丙氨酸→缬氨酸 (Ala→ Val) ,宣威农民肺癌患者中有 1例于 2 47位密码子突变 ,由 AAC→ TTC,导致氨基酸发生改变 ,由天冬酰氨→苯丙氨酸 (Asn→ Phe)。结论　 p5 3突变在云锡矿工肺癌和宣威农民肺癌中较常发生 ,并发现两例新的突变位点。 p5 3突变与病人的年龄、性别、坑龄及组织学分类无明显联系     

运动对骨骼－肌腱界面愈合组织的影响

利用纽西兰兔作体内实验，来分析手术后石膏固定，笼内活动，连续被动性活动三种不同的复健方法对肌腱－骨骼间界面愈合组织的影响。结果显示在术后第１５天愈合组织所能承受的最大张力，笼内活动组为０．８７５ｋｇ，石膏固定组为２．０１４ｋｇ，是笼内活动组的２３１．５２％，而连续被动性活动组为２．６３８ｋｇ，是笼内活动组的２６１．３５％。石膏固定组虽有很高的张力，但关节内粘连也较多。     

喉癌中EGFR基因扩增、表达及临床意义

目的研究喉癌中表皮生长因子受体(EGFR)基因的扩增、表达,探讨其在喉癌发生、发展中的作用及临床意义。方法采用差异PCR(differential PCR)方法检测40例喉鳞状细胞癌及配对癌旁正常组织中EGFR基因的扩增(即基因拷贝数增加);应用RT-PCR方法检测EGFR mRNA水平;应用SPSS13.0软件对数据进行统计学分析。结果喉癌组织中有13例(占32.5%)EGFR基因拷贝数增加,癌旁对照组中则未检测到(χ2=15.537,P<0.005);喉癌组织中EGFR mRNA平均积分光密度为872.356±62.340,癌旁对照组为346.425±57.380(t=5.959,P<0.001);喉癌组织分化程度越低,病理分期越晚,EGFR基因扩增和mRNA表达水平越高(P<0.05)。结论喉癌中EGFR基因在DNA水平上的扩增是EGFR mRNA过表达的原因之一,EGFR的扩增和过表达在喉癌的发生、进展中发挥一定作用。     

Quantitative real-time PCR detection of adenovirus in clinical blood specimens: A comparison of plasma, whole blood and peripheral blood mononuclear cells

BACKGROUND: Detection and quantification of adenovirus (ADV) in peripheral blood specimens has become an increasingly important tool in the management of immunosuppressed patients. Investigators have described the use of whole blood (WB), peripheral blood mononuclear cells (PBMC), serum and plasma but no studies have compared the utility of these different sample types for use in a clinical diagnostic assay. OBJECTIVES: To determine the optimal blood compartment for quantitative real-time measurement of adenovirus in peripheral blood specimens. STUDY DESIGN: WB, PBMC, and plasma representing 338 samples from 148 patients were tested for ADV by quantitative real-time PCR (qrt-PCR) and the results compared for concordance of both qualitative sensitivity and viral load among positive specimens. RESULTS: There was no significant difference in qualitative sensitivity among the three tested specimen types. Quantitative values of WB and plasma were similar and tended to be greater than those found in PBMC samples. Comparison of consecutive positive samples within individual patients showed that viral loads tracked similarly over time, irrespective of the sample type tested. CONCLUSION: While WB and plasma do not offer a significant increase in sensitivity over PBMC, they may offer benefits in terms of reduced processing costs and laboratory turn around time.     

阴道炎患者HPV6/11-DNA定量分析

目的：了解女性阴道炎患者人乳头瘤病毒6／11型（HPV6／11）的感染情况。方法：荧光定量聚合酶链反应（FQ－PCR）技术检测258例门诊患者阴道分泌物中HPV6／11的病毒拷贝数。结果：共检出HPV6／11阳性者94例,阳性率36．43％,拷贝数在10^5以上者78例,占阳性者中82．98％,40岁以上年龄组阳性率高且病毒复制量均在10^5以上。结论：（1）中老年阴道炎患者就诊晚,感染重。（2）FQ－PCR检测HPV敏感,快速,准确,特别是其定量特点对临床很有意义。     

EFFECT OF VL AND VH CONSENSUS SEQUENCE-SPECIFIC PRIMERS ON THE BINDING AND EXPRESSION OF A MINI- MOLECULE ANTIBODY DIRECTED TOWARDS HUMAN GASTRIC CANCER

Objective. To construct ScFv and Fab from murine anti-gastric cancer monoclonal antibody(mAb) 3H11.Methods. At first,3H11 ScFv and Fab were constructed with Ⅴ genes PCR amplified by degenerate primers for FR1 .The bacterial expressed 3H11 Ab fragments showed no antigen binding activity.Then,phage antibody library and random mutated library were constructed from 3H11 hybfidoma cells and panning selection was performed. Again the i-dentification of positive clone was failed. Finally the N-terminal sequences of Ⅴ regions were resumed to 3H11 original sequences by site-directed mutagenesis via PCR.Restdts. Binding activity to gastric cancer cells was detected only from N-terminal sequence corrected 3H11 ScFv and Fab, though the expression of the Ab fragments was not affected. Correction of either VL or VH N-terminal se-quences could partially resume the antigen binding activity. Conclusion. Sequence changes of Ⅴ region N terminal introduced by PCR may seriously affect antigen binding without affecting the expression of antibody.