应用创造酶切位点PCR-RFLP检测乙醇脱氢酶2基因多态性

目的建立简便易行经济适用的乙醇脱氢酶2(ADH2)基因单核苷酸多态性检测方法。方法根据单碱基突变位点的碱基替代情况设计引物,其中一条引物根据突变位点邻近序列设计,人为引入错配碱基,使引物3′端和单碱基突变的一种突变型在PCR扩增后形成一个酶切位点,并应用相应内切酶进行酶切鉴定,通过PCR-RFLP技术判断ADH2基因SNP位点多态性。结果设计一对特异引物并PCR扩增含ADH2多态位点的DNA片段,使用限制性内切酶Bsh1236I酶切判断ADH2位点多态性,PCR和酶切效果均较好。结论应用创造酶切位点PCR-RFLP原理建立的ADH2多态位点检测方法具备简便、经济、快速的特点。     

维吾尔族人群代谢性疾病与ApoEε2等位基因

目的探讨维吾尔族代谢性疾病与载脂蛋白Eε2等位基因(ApoEε2)的关系。方法收集2006年1月-2006年12月在新疆医科大学附属中医医院和宝科达医院健康体检的386名维吾尔族人,采集血液标本,检测血液生化指标以及ApoE基因型。结果新疆维吾尔族ApoEε2等位基因的携带率为31.9%(123/386);与非携带组比较,除了高密度脂蛋白-胆固醇升高外,ApoEε2等位基因携带组的其余指标均有下降趋势,其中收缩压、舒张压及低密度脂蛋白差异有统计学意义(P0.05);与非携带组比较,携带者代谢性疾病检出率降低,尤其是对高胆固醇血症和高血压影响较大,其中高血压检出率差异,P=0.058。结论ApoEε2等位基因携带者血脂水平及代谢性疾病的检出率降低,是代谢性疾病的保护因素。     

绞股蓝属植物及其混淆品乌蔹莓的PCR-RFLP鉴别研究

目的：寻找简便、可重复的分子标记方法对绞股蓝属Gynostemma植物及其混淆品乌蔹莓Cayratia japonica进行鉴别。方法：对7种常见药用绞股蓝属植物及其混淆品乌蔹莓的6个cpDNA片段进行PCR 扩增，再利用TaqⅠ，HpaⅡ，EcoRⅠ，RsaⅠ，HhaⅠ，HindⅢ等6种限制性内切酶分别对扩增片段进行消化。结果：36种DNA片段/内切酶组合中，trnK1f -trnK2r片段与RsaⅠ组合可将乌蔹莓从绞股蓝属植物中区分出来，产物清晰、结果稳定。结论：PCR-RFLP分析方法可有效区分常见绞股蓝属植物与其混淆品乌蔹莓。     

Polymorphism of CYP1A1 gene variants rs4646903 and rs1048943 relation to the incidence of cervical cancer in Chhattisgarh

Cytochrome P450 CYP1A1 is a phase 1 xenobiotic metabolizing enzyme involved in the metabolism of toxins, endogenous hormones and pharmaceutical drugs. It is therefore possible that polymorphism of CYP1A1 gene producing functional changes in the enzyme may be susceptible factors in cervical carcinogenesis. This study was aimed to look association of CYP1A1 m1 (T > C) and m2 (A > G) gene polymorphisms in Chhattisgarh population. In this case-control study, we analyzed leukocyte DNA from a total of 200 subjects form Chhattisgarh (100 cases and 100 controls). All subjects were genotyped for CYP1A1 m1 (T > C) and m2 (A > G) using PCR-RFLP with statistical analysis by using SPSS version 16.0 and VassarStats (online). Among the two gene variants rs4646903 (T > C) and rs1048943 (A > G), individuals with AG and GG genotypes of CYP1A1 m2 polymorphism have significantly higher and increased risk of cervical cancer (OR = 2.0, 95%CI = 1.04-3.84, p = 0.035; OR = 62.9, 95%CI = 3.72-1063.83, p = 0.004 respectively) and the association of CYP1A1 m1 polymorphism did not show any significant relationship with cervical cancer patients (p = 0.23). The ‘G’ allele showed strong association with the disease (p < 0.0001). Thus, CYP1A1 m2 polymorphism showed an increased risk in the population leading to cervical cancer. Our study suggested that the presence of ‘C’ allele of rs4646903 (T > C) showed no risk and ‘G’ allele of rs1048943 (A > G) might be a leading allele to cause increased cervical cancer susceptibility due to significant association of CYP1A1 m2 gene polymorphism.     

中国汉族人群hGSTA1基因C-69T多态性分析

目的　研究中国汉族人群中人谷胱甘肽S 转移酶(hGSTA1 )C-69T基因多态性的分布。方法1 4 0例血样本来自中国2 5个省份的汉族人口,用聚合酶链反应 限制性片段长度多态性方法检测hGSTA1 69位点的变异。结果　中国汉族人群GSTA1基因 69位点的野生型纯合子(CC)基因型的分布频率为75 .0 %,突变型纯合子(TT)基因型为0 .7%,杂合子(CT)基因型为2 4 .3 %;C及T两种等位基因的频率分别为87.1 %及1 2 .9%。结论中国汉族人群GSTA1基因呈多态性分布,其等位基因和基因型频率不同于其他种族     

川贝母PCR-RFLP法鉴别检验能力验证活动分析

目的：考察相关实验室执行《中华人民共和国药典》川贝母PCR-RFLP法鉴别检验项目的技术能力,促进分子生物学技术在中药检定中的广泛应用。方法：测试样品为粉末,对样品的均匀性和稳定性进行评价,样品种类分为阳性和阴性两种,采用随机单盲方式分配至参与能力验证的实验室。各实验室按照作业指导书进行检测,并以规定格式书写原始记录。检测结果与样品性质一致的实验室为满意。对各实验室的反馈结果进行汇总和分析。结果：最终报名参加的实验室总计50家,能力验证结果为满意的26家,占52%,不满意24家,占48%。其中3家试验室未按要求返回结果,判定为不满意。分别统计不同类型参加实验室的满意率,地市级食药检机构满意率最高,为70.6%。结论：总体满意率偏低,川贝母PCR-RFLP法鉴别检验项目的整体技术水平有待提高。     

过氧化物增值激活受体γ2基因多态性对高血糖人群膳食干预效果的影响

目的 探讨过氧化物增值激活受体(PPAR)γ2基因多态性对高血糖人群膳食干预效果的影响.方法 在南京主城区筛选241位高血糖汉族居民,设立干预组(129人)和对照组(112人),干预组给予粗杂粮馒头同时进行健康教育,对照组只进行健康教育,6个月进行一次体检,干预期为1 a.结果 干预组体质指数(BMI)、腰臀比(WHR)、空腹血糖(FBG)、胆固醇(TC)、甘油三酯(TG)比干预前降低(P<0.05),高密度脂蛋白胆固醇(HDL-C)升高(P<0.05);干预组干预后BMI、WHR、FBG、TC、TG低于对照组(P<0.05),HDL-C高于对照组(P<0.05).无论Pro/Pro基因型还是Pro/Ala基因型人群,干预后体重、BMI、FBG、TC、TG均比干预前降低(P<0.05);Pro/Ala基因型人群体重、BMI、TG的下降幅度大于Pro/Pro型人群(P<0.05).结论 复合式膳食干预对高血糖人群具有良好的效果;Pro/Ala基因型人群干预效果好于Pro/Pro基因型人群.     

基于trnL-trnF序列分析的何首乌PCR-RFLP分子鉴别

目的:建立何首乌(Fallopia multiflora)DNA分子鉴别技术。方法:对何首乌与其近缘种及其混淆品的trnL-trnF(trnL基因和trnF基因间隔区)序列进行比较分析。结果:何首乌与其近缘种及其混淆品的trnL-trnF差异率为2.1%～22%,何首乌种内各居群间trnL-trnF差异率为0%～1.5%。基于何首乌与其近缘种及其混淆品的trnL-trnF序列的差异,找出一个位于trnL5'-trnL3'间区的何首乌特征性Xba I酶切位点(T↓CTAGA),用Xba I酶对不同采集地的何首乌样品trnL-trnF序列扩增产物酶切后均得到含约804～819 bp和256 bp两个片段的PCR-RFLP图谱,而其混淆品trnL-trnF序列扩增产物因不能被Xba I酶切,图谱呈单一条带。结论:利用建立的PCR-RFLP方法可以很好地区分何首乌及其混淆品植物。     

Asociación entre polimorfismos de los genes POLG y XRCC1 y aparición de queratocono entre pacientes egipcios

BackgroundKeratoconus is a progressive disorder distinguished by thinning of the corneal tissue and bulging forward into a cone-shaped fashion. Yet its etiology, which is multifactorial, despite intensive research remains elusive. Corneal exposure a reactive oxygen species causing oxidative DNA damage has been reported to be associated with KC and therefore suggesting that DNA base excision repair mechanism might lie behind the pathogenesis of the disease.MethodsWe studied the association of three variants in two BER genes (XRCC1 and POLG) and QC occurrence in a cohort of patients from Egypt. Genotyping of the three variants was performed using PCR and restriction enzymes analysis.ResultsWe observed that A allele and A/A genotype of the c.1196A>G variant in the XRCC1 gene were significantly associated with increased KC occurrence while the G allele was associated with decreased KC occurrence. Similarly, the A/A genotype of the c.–1370T>A polymorphism in the POLG gene and the A allele were associated with increased occurrence of KC, while T/A genotype and the T allele were accompanied with decreased occurrence of KC. On the other hand, no association was observed between the c.580C>T variant in the XRCC1 gene and KC occurrence among the studied group of patients.ConclusionOur results suggest that c.1196A>G variant of the XRCC1 and c.–1370T>A variant of the POLG gene may be involved in KC pathogenesis and might be considered as a genetic risk factors of the disease among Egyptian population.     

Comparison of DNA fingerprinting by PFGE and PCR-RFLP of the coagulase gene to distinguish MRSA isolates

Staphylococcus aureus isolates were collected from epidemiologically unrelated clinical sources in Japan between 1991 and 1993. A total of 40 isolates, five each of eight coagulase types, were analysed by polymerase chain reaction (PCR) of the coagulase gene, PCR-restriction fragment length polymorphism (RFLP) after AluI digestion, and pulsedfield gel electrophoresis (PFGE) of chromosomal DNA after SmaI digestion. The efficiency of discrimination among the isolates increased in the order of PCR < PCR-RFLP < PFGE, yielding five, 13 and 31 different types, respectively. To assess the clinical use of these methods, 42 additional methicillin-resistant S. aureus (MRSA) isolates collected from 27 inpatients in a hospital were analysed. PFGE and PCR-RFLP were able to discriminate 11 and four types, respectively. PFGE analysis detected cross-infection between four postoperative patients in an intensive-care unit, and in six neonates in intensive care. We conclude that of the three methods tested, PFGE analysis currently allows the most effective discrimination of MRSA strains.