ACE基因rs4353,rs4461142和rs8066114位点多态性与心肌梗死的相关性分析

目的 探讨陕西地区汉族人群中,血管紧张素转换酶(ACE)基因多态性与心肌梗死(MI)的相关性。方法 选择2013年1月～2017年1月住院治疗的346例急性心肌梗死(AMI)患者纳入研究记为MI组,另选择300例健康志愿者作为对照组,采用ELISA法检测血清ACE水平,采用聚合酶链反应-限制性片段长度多态性法检测ACE基因rs4353,rs4461142和rs8066114多态性,并收集患者临床资料进行分组及统计学分析。结果 MI组与对照组比较,两组血清ACE水平34.7±13.8U/L vs 30.6±12.0 U/L,差异具有统计学意义(t=3.998,P<0.05)。对于对照组患者,ACE基因rs4353和rs4461142位点不同基因型血清ACE水平差异具有统计学意义(F=40.860,3.382,均P<0.05); rs8066114位点不同基因型血清ACE水平差异无统计学意义(F=0.176,P>0.05)。MI组和对照组比较,ACE基因rs4353位点基因型和等位基因分布频率差异具有统计学意义(χ²=8.146,7.585,均P<0.05),两组rs4461142和rs8066114位点基因型和等位基因分布频率差异均无统计学意义(χ²=2.263～4.016,均P>0.05)。多因素Logistic回归分析结果显示ACE基因rs4353位点A等位基因是MI危险因素,可显著增加MI发生风险(OR=1.620,95%CI 1.217～4.723,P=0.035)。结论 ACE基因rs4353位点多态性可能与MI易感性相关。     

高分辨率熔解曲线技术检测冠心病患者APOB、APOC-Ⅰ和LDLR基因rs693、rs4420638、rs688位点多态性

目的建立APOB基因rs693位点、LDLR基因rs688位点和APOC-Ⅰ基因rs4420638位点单核苷酸多态性(SNP)的聚合酶链式反应-高分辨率熔解曲线分析(PCR-HRM)的检测方法,并探讨该3个SNP位点与冠心病(CHD)易感的相关性。方法针对rs693、rs688和rs4420638 3个位点分别设计引物,建立PCR-HRM分子诊断方法,并通过直接测序进行验证。对311例CHD患者和300例体检健康者进行病例-对照研究,分析其与CHD易感性及血脂指标(TC、TG、HDL-C、LDL-C)的关系。结果rs693和rs4420638位点的基因频率和基因型频率在CHD组与对照组之间差异有统计学意义(P0.05)。rs693位点CT基因型降低CHD患病风险(OR=0.448,95%CI:0.246~0.817,P=0.009);rs4420638位点AG基因型增加CHD患病风险(OR=2.140,95%CI:1.241~3.688,P=0.006)。rs688(CT)和rs4420638(AG)位点组成的单倍型CG增加CHD患病风险(OR=1.715,95%CI:1.091~2.697,P=0.018)。病例组rs4420638位点AG基因型患者的血清TC水平显著高于AA基因型(F=4.281,P=0.040)。结论成功建立了rs693、rs688和rs4420638 3个SNP位点的PCR-HRM基因分型方法。rs693和rs4420638位点以及单倍型CG与中国兰州地区汉族人群CHD易感性有关,rs4420638位点联合血清TC水平检测可预警CHD发生。     

血管紧张素转换酶和载脂蛋白E基因单核苷酸多态性与维吾尔族糖尿病相关性

目的探讨新疆维吾尔族血管紧张素转换酶(angiotensin-converting enzyme,ACE)、载脂蛋白E(apolipoprotein E,ApoE)基因单核苷酸多态性位点(single nucleotide polymorphism site,SNPs)与糖尿病的关系。方法采用整群抽样法抽取喀什地区不同经济水平的3个乡镇/街道居民772例,根据是否有糖尿病分为糖尿病组112例和对照组660例,应用HaPloview 4.2软件查找ACE基因、ApoE基因标签SNPs,采用PCR-高分辨率熔点曲线对ACE基因、ApoE基因SNPs标签位点进行基因分型,比较2组ACE、ApoE基因标签SNPs基因型、等位基因分布频率,分析标签位点间的连锁不平衡和单倍体型。结果 2组ACE基因6个SNPs和ApoE基因2个SNPs基因型和等位基因频率分布比较差异均无统计学意义(P0.05);ACE基因rs4353和rs4461142位点存在较弱的连锁不平衡(D’=0.80,r~2=0.650),ApoE基因rs405509和rs769450位点存在连锁不平衡(D’=0.92,r~2=0.492);糖尿病组ApoE基因rs405509和rs769450构成的A-C单倍体型等位基因频率(33.6%)明显高于对照组(26.6%)(P0.05),ACE基因rs4353和rs4461142构成的单倍体型等位基因频率与对照组比较差异均无统计学意义(P0.05)。结论 ACE基因6个标签位点SNPs可能与糖尿病发病无关;ApoE基因2个标签位点SNPs基因型及等位基因频率与糖尿病发生无关,其构成的单倍体型可能与糖尿病发生有一定关系。     

高分辨率熔解技术检测冠心病患者TNF-α及其受体基因多态性

目的用高分辨率熔解(HRM)技术检测兰州地区汉族人群肿瘤坏死因子α(TNF-α)及其受体基因单核苷酸多态性(SNP),探讨其与冠心病(CHD)易感性及血脂指标的相关性。方法建立TNFA-238GA、TNFA-308GA、TNFA-857CT、TNFA-863CA、TNFR1-383AC和TNFR2-exonTG 6个SNP位点的PCR-HRM检测方法,并对207例CHD患者和274例健康对照进行病例-对照研究,分析其与CHD易感性及血脂指标的关系。结果仅TNFA-863CA位点的基因频率和基因型频率在CHD组和对照组之间差异具有统计学意义(P=0.020,0.013),性别分层后仅存在于男性患者(P=0.026,0.018),CA基因型明显增加CHD患病风险(OR=1.999,P=0.029)。针对TNF-α基因的单倍型分析显示TNFA-238G/-308G/-857C/-863A(GGCA)单倍型明显增加CHD患病风险(OR=1.695,P=0.016)。病例组的血脂分析显示TNFR2-exon位点与CHD患者的高密度脂蛋白胆固醇(HDL-C)水平有关(F=5.126,P=0.024)。结论 TNFA-863CA位点和单倍型TNFA-GGCA与兰州地区汉族人群CHD易感性相关,TNFR2-exonTG位点与CHD患者HDL-C水平相关联。     

汉族健康人群血管紧张素转换酶多态性及其血清水平的研究

目的调查汉族健康人群血管紧张素转换酶(ACE)基因的单核苷酸多态性(SNP)分布与血清ACE水平的相关性。方法提取132例健康个体外周血有核细胞DNA,应用荧光标记单碱基延伸分型技术及寡核苷酸微阵列芯片杂交技术检测ACE基因的2个标签SNP(tag SNP)rs4353和rs4305;应用单一试剂速率法检测血清ACE水平。结果汉族健康人群ACE基因rs4353多态性中AA型占23.4%,AG型占47.7%,GG型占28.9%;rs4305多态性中AA型占10.1%,AG型占49.6%,GG型占40.3%。rs4353的AA和AG基因型血清ACE水平明显高于GG基因型;rs4305的AG基因型血清ACE水平明显高于GG基因型(P0.05)。结论 ACE基因2个tag SNP的基因型与血清ACE水平密切相关。     

胰岛素样生长因子Ⅱ及其受体基因与汉族人高度近视的关联研究

目的探讨胰岛素样生长因子Ⅱ(IGF2)、胰岛素样生长因子Ⅱ受体(IGF2R)基因与中国汉族人高度近视(HM)发病之间的相关性。方法采用Sequenom MassARRAY方法检测基因的12个单核苷酸多态性(SNPs),对基因分型测试数据进行了χ~2分析和利用Haploview软件进行连锁不平衡分析。结果 IGF2基因的SNP rs1003483与HM具有显著相关性(等位基因P=0. 0023)。未发现IGF2R基因的单核苷酸多态性与HM相关。结论胰岛素样生长因子Ⅱ基因变异会增加中国汉族人群高度近视易感性。     

QKI基因多态位点与四川地区汉族人群冠心病的相关性分析

目的研究QKI基因单核苷酸多态性(SNP)位点与四川地区汉族人群冠心病(coronary heart disease,CHD)发病的相关性。方法入选570例CHD患者和735例正常对照(NC)样本,通过病例-对照关联研究方法,选择QKI基因上4个标签SNP(rs7756185、rs6941513、rs7745161、rs7751144),以单碱基延伸(SNa Pshot)方法进行基因分型,并分析其与CHD的相关性。结果 QKI基因4个SNP位点的基因型分布均符合Hardy-Weinberg平衡(P0.05)。其等位基因频率在CHD组与NC组之间差异有统计学意义(均P0.01)。通过单倍体型分析发现,这4个SNP位点处于同一个连锁不平衡区域,其风险单倍体型TGGG可以增加CHD易感性0.25倍(P=0.0051),而保护型的单倍体型GACA可降低CHD患病风险31%(P=0.0003)。结论 QKI基因多态性位点与四川地区汉族人群CHD发病显著相关,其保护型等位基因可降低CHD的易感性。     

ATG5基因启动子区域单核苷酸多态性与急性心肌梗死的相关性分析

目的探讨自噬相关蛋白5(autophagy-related protein,ATG5)基因启动子序列单核苷酸多态性(single nucleotide polymorphisms,SNPs)与急性心肌梗死(acute myocardial infarction,AMI)的关系。方法采用PCR和Sanger基因测序法检测ATG5基因启动子序列SNPs;通过卡方检验、Logistic回归分析及单倍型分析法对378例AMI患者和374例健康对照人群的SNPs进行分型和关联分析。结果发现ATG5基因启动子序列中的2个SNPs:rs506027[OR=1.4,95%CI(0.6～3.0),P=0.411]和rs510432[OR=1.6,95%CI(0.7～3.4),P=0.275]并未增加AMI的易感性,且未发现该2个SNPs与AMI相关的单倍型。结论ATG5基因启动子多态性与AMI易感性无关。     

EZH2基因多态性与结直肠癌遗传易感性的关联性研究

摘要：目的：探讨EZH2(en hancer of zeste homolog 2)基因单核苷酸多态性（SNPs)与结直肠癌（CRC）遗传易感性的相关性。 方法：用病例-对照方法分析EZH2基因的2个SNPs位点（rs887569与rs1880357）在中南地区汉族人群中的分布。提取96例CRC患者与100例体检健康者的外周血DNA;用聚合酶链式反应-限制性片段长度多态性法（PCR-RFLP）检测EZH2基因型并计算相应等位基因频率;DNA测序验证基因分型结果。 结果：CRC组与健康对照组年龄分布（t=0.693,P=0.489）与性别构成（χ²=0.403,P=0.526）差异无统计学意义。CRC组与健康对照组EZH2基因rs887569位点与rs1880357位点基因频率分布均符合Hardy-Weinberg平衡（P均>0.05）,证明所观察的样本具有群体代表性。EZH2基因rs887569位点TT、CC和TC基因型（χ²=1.531,P=0.465）及rs1880357位点GG、CC和GC基因型（χ²=0.670,P=0.413）在CRC患者和健康对照者间分布差异无统计学意义。对等位基因而言,rs887569位点C>T与rs1880357位点C>G等位基因分布在CRC患者与健康对照者间差异亦无统计学意义（OR=0.875,95%CI:0.488~1.566,P=0.667;OR=0.793,95%CI:0.403~1.563,P=0.503）。 结论：EZH2基因rs887569与rs1880357 SNPs位点多态性可能与CRC发病风险无关。     

LTA基因单核苷酸多态位点与胃癌易感性的研究

目的通过对江西地区汉族胃癌患者和正常人群中淋巴毒素-α（LTA）基因单核苷酸多态（single nucleotide polymorphism,SNP）位点rs909253基因型的检测,探讨rs909253位点与胃癌患者易感性的关系。方法利用Sequenom-MassArray-IPLEX检测194例胃癌和250例对照LTA基因多态位点rs909253的基因分型,并利用非条件logistic回归对检测结果进行相关统计。结果 LTA rs909253多态位点GG、GA、AA三种基因型在胃癌的频率分别为26.4%、52.0%和21.6%,与对照组（35.6%、46.4%和18.0%）相比,统计学上无显著意义（χ2=4.403,P=0.111）;但胃癌组和正常组的等位基因频率分布差异临近显著,P值达0.059;对年龄和性别进行校正后,携带等位基因G的胃癌发病风险有所增加（OR=1.315,95%CI：1.001~1.728）。结论江西汉族人群胃癌的遗传易感性可能与LTA基因rs909253位点的单核苷酸多态性有关,G等位基因为其发病的危险因素。     

Genetic polymorphisms in sepsis

The number of genetic polymorphisms shown to play a role in sepsis continues to increase. At the same time, platforms for genetic sequencing and expression analysis are being refined, allowing unprecedented data generation. International databases may soon facilitate synchrony of genotypic and phenotypic data using enormous numbers of septic patients. If this occurs, 2 strategies for investigating polymorphisms in sepsis are likely to gain favor. In the first strategy, sepsis will continue to be viewed as a single entity. High-throughput genetic techniques will be used to evaluate numerous polymorphisms, each with fractional disease responsibility. Nongenetic variables, such as pathogen characteristics, underlying host medical conditions, and type and timing of resuscitation, will be considered cofactors. Using this approach, principal components that predict susceptibility to and outcomes during sepsis are likely to be identified. In the second strategy, sepsis will be divided into subtypes based on the concentration of specific variables. Categories will be based on features like the presence or absence of specific polymorphisms, gram-positive or gram-negative staining of causative organisms, age and comorbid conditions of the host, recent administration of chemotherapeutic agents, and hospital setting (ie, community vs teaching institution). Each category will be used to create homogenous sepsis subgroups for detailed evaluation. This approach will increase the odds of finding single dominant factors responsible for predilection and/or outcome within well-defined groups among those with sepsis. Several elements will be essential for the success of both these strategies. Firstly, databases that are extremely detailed will have to be generated. Secondly, better clinical information technology systems will be needed to facilitate large-scale phenotyping. Thirdly, standardization of protocols will need to take place to ensure uniformity of data sets. If the rapid advances in technology and informatics continue, they may catalyze paradigm shifts with regard to how clinicians address sepsis. Clinicians may change their focus from aggressive uniform treatment strategies to rapid stratification and subcategorization, with subsequent aggressive targeted therapeutic interventions. Advances in technology have the potential to change our primary goal in sepsis from rapid treatment to prevention for those most at risk. The cost savings to the US health care systems from such changes could be substantial.     

Genetic polymorphisms and sepsis

Sepsis is a polygenic and complex syndrome that is initiated by infection and is characterized by a systemic inflammatory response. Genetic polymorphisms in the immune response to infection have been shown to be associated with clinical outcomes. Functional and association studies involving genetic polymorphisms in essential genes, including Toll-like receptors, cytokines, and coagulation factors, have provided important insights into the mechanisms involved in the pathogenesis of sepsis-induced organ dysfunction. The advancement of high-throughput single nucleotide polymorphism (SNP) genotyping will provide valuable information on the interaction of multiple allelic variants and clinical outcome. More precise categorization of patients based on genetic background is likely to lead to individualized targeted treatment. Future therapeutic trials as well as actual treatment regimens for patients with sepsis are likely to be designed to target specific genotypes and associated cellular responses, maximizing clinical response and patient safety.     

Genomic polymorphisms in sepsis

OBJECTIVE: This article aims to review all relevant genetic polymorphism studies that may contribute to the pathogenesis of sepsis with emphasis on polymorphisms of the innate immunity, pro- and anti-inflammatory cytokines, and coagulation mediators. DATA SOURCE: Published articles reporting on studies of associations between genetic polymorphisms, sepsis, septic shock, and other relevant infectious disease models. DATA ANALYSIS: Research into the pathogenesis of sepsis has led to the development of many potential therapeutic strategies. Several therapeutic agents and treatment modalities have been shown to decrease mortality rates in large, prospective, and randomized clinical trials. However, although these advances have resulted in improved survival for certain patient populations, the overall mortality rate for septic patients remains high. With the rapid development of molecular and genetic techniques, substantial interests have developed in using genomic information to define disease-mediating genetic variants in sepsis. Combined with microarray technology, it is anticipated in the near future that one will be able to tailor drug selection and dosage and predict outcome by correlating genetic profile with disease presentation. Numerous genetic association studies in sepsis have already been reported and more are likely to be published. CONCLUSIONS: Although studies examined in this review are of small heterogeneous populations, the identification of strong associations between certain genetic polymorphisms and increased mortality rate or susceptibility to severe sepsis is intriguing and supports further research using this approach. The establishment of these associations does not equal causation, and further research is required in both genetic and molecular aspect of sepsis.     

Y chromosome polymorphisms in medicine

Ninety‐five percent of the length of the human Y chromosome is inherited as a single block in linkage from father to male offspring as a haploid entity. Thus, the Y chromosome represents an invaluable record of all mutations that have occurred along male lineages throughout evolution. For this reason, Y chromosomal DNA variation has been mainly used for investigations on human evolution and for forensic purposes or paternity analysis. Recently, Y chromosomal polymorphisms have been applied in molecular medicine from the perspective of male‐specific (spermatogenic failure, testis and prostate cancer) and prevalently male‐associated (hypertension, autism) diseases. The absence of recombination on the MSY (male‐specific Y) region means that polymorphisms, located in this region, are in tight association with potential functional variations associated with Y‐linked phenotypes. Thus, an indirect way to explore if Y chromosome genes are involved in the etiology of a specific disease is the definition of Y chromosome haplogroups in patients versus disease‐free and/or the general population. Data on patients with reduced sperm count and prostate cancer indicate that the ‘at risk Y haplogroup’ may be different in different populations. The situation is rather contradictory for other male‐specific or male‐associated diseases and further multicenter – possibly multiethnic – studies are needed.     

Genotyping Parkinson disease-associated mitochondrial polymorphisms

OBJECTIVE: The purpose of this study was to establish a system for rapidly detecting single nucleotide polymorphisms (SNPs) in mitochondrial DNA (mtDNA) using hybridization probes and melting temperature (T(m)) analysis. This technology should prove useful for population-based studies on the interaction between genetic factors and environmental exposures and the risk of Parkinson disease (PD). METHODS: Mitochondrial DNA (mtDNA) was extracted from whole blood. Rapid polymerase chain reaction (PCR) and melting curve analyses were performed with primers and fluorochrome-labeled probes on a LightCycler (Roche Molecular Biochemical, Mannheim, Germany). Genotyping of 10 SNPs in 15 subjects was based on the analysis of allele-specific T(m) of detection probes. The results of melting curve analyses were verified by sequencing all 150 PCR products. RESULTS: Real-time monitoring showed optimal PCR amplification of each mtDNA fragment. The nucleotide changes at positions 1719, 4580, 7028, 8251, 9055, 10398, 12308, 13368, 13708, and 16391 from wild-type to mutant genotype resulted in 6.51, 8.29, 3.26, 7.82, 4.79, 2.84, 2.73, 9.04, 8.53, and 9.52 degrees C declines in T(m) of the detection probes, respectively. Genotyping of all 150 samples was verified by 100% correspondence with the results of sequencing. Fourteen subjects were haplogrouped by combining results for all 10 SNPs. CONCLUSION: A rapid and reliable detection system for identifying mitochondrial polymorphisms and haplotypes was developed based on hybridization probe technology. This method may be suitable for mitochondrial genotyping of samples from large-scale epidemiology studies, and may prove useful for exploring the molecular etiopathogenesis of PD, identifying markers of genetic susceptibility, and protecting susceptible individuals from PD.     

Genetic polymorphisms of drug metabolism

The molecular mechanisms of 3 genetic polymorphisms of drug metabolism have been studied at the level of enzyme activity, enzyme protein and RNA/DNA. As regards debrisoquine/sparteine polymorphism, cytochrome P-450IID6 was absent in livers of poor metabolizers; aberrant splicing of premRNA of P-450IID6 may be responsible for this. Moreover, 3 mutant alleles of the P-450IID6 locus on chromosome 22 associated with the poor metabolizer phenotype were identified by Southern analysis of leucocyte DNA. The presence of 2 identified mutant alleles allowed the prediction of the phenotype in approximately 25% of poor metabolizers. The additional gene-inactivating mutations which are operative in the remainder of poor metabolizers are now being studied. Regarding mephenytoin polymorphism, although the deficient reaction, S-mephenytoin 4'-hydroxylation, has been well defined in human liver microsomes, the mechanism of this polymorphism remains unclear. All antibodies prepared to date against cytochrome P-450 fractions with this activity recognize several structurally similar enzymes and several cDNAs related to these enzymes have been isolated and expressed in heterologous systems. However, which isozyme is affected by this polymorphism is not known. As regards N-acetylation polymorphism, N-acetyltransferases have been purified from human liver, specific antibodies prepared; it was observed that immunoreactive N-acetyltransferase is decreased or undetectable in liver of "slow acetylators". Two genes that encode functional N-acetyltransferase were characterized. The product of one of these genes has identical activity and characteristics as the polymorphic liver enzyme. Cloned DNA from rapid and slow acetylator individuals has been analyzed to identify the structural or regulatory defect that causes deficient N-acetyltransferase.