多重SNP基因分型和基因表达检测法的构建及其在冠心病易感性和诊断中的应用研究
|
摘要
[目的]近年来我国CAD的患病率和死亡率呈逐年上升趋势。CAD是遗传因素和环境因素共同作用导致粥样硬化斑块在冠状动脉壁沉积而引起的。为了寻找与CAD相关的SNP位点和基因,本课题构建了多重SNP基因分型和多重基因表达检测方法,并对CAD患者和健康对照人群外周血白细胞的多个SNP位点和多个目标基因的表达水平进行了检测。
[方法](1)多重SNP基因分型的检测:提取外周血白细胞DNA,选择HapMap上公布的ADD1,PECAM-1,CRP,ecNOS,PC-1,SELL,GNB3,ACE,AT1R,AGT,ICAM-1,MTHFR和HL基因中国汉族人群的44个标签SNP,每个SNP设计3条引物(2条PCR扩增引物和1条延伸引物),通过PCR扩增、延伸及杂交,应用SNP stream基因分型系统对SNP位点进行基因分型。(2)多重基因表达水平的检测:提取外周血白细胞RNA,选择MTHFR,IL-8,ecNOS,TLR4,FOS,ID2,ADD1,ICAM-1,PECAM-1,CRP,LDLR,vWF,SELL,TNF-α等14个目标基因和β-actin、SRP14及18S-rRNA等3个管家基因,每个基因设计2条引物(1条反转录引物和1条PCR扩增引物),通过反转录和PCR扩增,应用高通量的GenomeLab GeXP分析仪检测17个基因的表达水平。(3)应用全自动生化分析仪检测血脂、CRP、ACE及Hcy水平。
[结果](1)构建的44个SNP分型成功32个,分型成功率为72.7%(32/44)。通过对583份DNA标本进行检测,32个SNP的平均样本检出率为98.3%。通过对中国汉族CAD患者和健康对照人群的32个SNP进行检测,结果显示CRP,ecNOS,tPc-1和ACE基因中的6个SNP的基因型频率和(或)等位基因频率在两组间存在统计学差异。根据连锁不平衡水平构建单倍型,结果显示ecNOS,PC-1,ACE基因的单倍型频率与CAD相关。实验发现CRP,ACE和MTHFR基因多态性分别与血清CRP,ACE和Hcy水平相关。(2)通过单重基因扩增、电泳,确认每个基因扩增产物的大小。结果显示扩增产物的大小与设计大小基本一致。根据每个基因扩增产物、质控基因和内标的片段大小,确认实验设计的每个基因扩增产物的特异性。在多重基因扩增中,根据电泳结果对外周血白细胞中高丰度表达基因的引物进行倍比稀释,确定了体系中每个基因的最佳引物浓度。通过对3个管家基因在两组之间表达水平的比较,发现以β-actin和SRP14基因的几何均数作为标准化因子可用以标准化本体系目标基因的表达。应用上述构建的14重基因表达检测方法对CAD患者和健康对照人群外周血白细胞RNA的基因表达水平进行检测,发现ecNOS,SELL,ADD1和MTHFR基因的表达水平在两组间存在统计学差异,其表达水平与部分SNP的基因型有关。
[结论]本实验构建的32重sNP基因分型和14重基因表达检测方法具有快速、准确、高通量、操作简单、微量化等优点。实验发现的与CAD相关的6个sNP位点和4个表达差异基因为心血管危险因素的预测和诊断提供了一种新思路,为心血管疾病寻找更有效的防治途径,早期预防和及时治疗提供理论指导。
[Objective]Coronary artery disease(CAD) is the leading cause of morbidity and mortality in recent years in China.It is a multifactorial disease caused by various genetic and environmental factors involved in accumulation of atherosclerosis in the walls of the coronary arteries.In order to find single nucleotide polymorphisms(SNP) and genes that were associated with CAD, multiplexed SNPs genotyping and genes expression methods were established in the current work.With the multiplexed methods,lots of tag SNPs and genes in peripheral blood white cells from CAD patients and normal control individuals were investigated.
[Methods](1) The method of multiplexed SNPs genotyping:DNA samples were extracted from peripheral blood white cells and 44 tag SNPs of 13 genes (ADD1,PECAM-1,CRP,ecNOS,PC-1,SELL,GNB3,ACE,AT1R,AGT, ICAM-1,MTHFR and HL) that were listed in the International Haplotype Map Project Database for CHB(Chinese Han nationality in Beijing) were selected in the current work.Three primers were designed for each SNP(two for polymerase chain reaction(PCR) and one for extension reaction).After PCR, extension and hybridization,genotypes of SNPs were investigated with the GenomeLab SNPstream genotyping system.(2) The method of multiplexed genes expression:RNA samples were extracted from peripheral blood white cells and two primers(one for reverse transcription and one for PCR) were designed for each of the 14 target genes(MTHFR,IL-8,ecNOS,TLR4,FOS, ID2,ADD1,ICAM-1,PECAM-1,CRP,LDLR,vWF,SELL and TNF-α) and 3 house keeping genes(β-actin,SRP14 and 18S-rRNA).After reverse transcription and PCR,expression levels of the 17 genes were detected with high-throughput GenomeLab GeXP Genetic Analysis System.(3) The levels of serum lipid,CRP,ACE and Hcy were detected with automatic biochemical analysis system.
[Result](1) Thirty two SNPs were genotyped and the rate of succeed SNP genotyping was 72.7%(32/44).The average genotyped rate of the 32 SNPs was 98.3%in the test of 583 DNA samples.With the 32 plexes SNPs genotyping method,DNA samples of CAD patients and normal control individuals from the local ethnic Han Chinese population were investigated.The results showed that the genotype and/or allele frequencies of 6 tag SNPs in CRP,ecNOS,PC-1 and ACE genes were significantly different in both groups.According the linkage-disequilibrium levels of SNPs,the haplotypes were constructed and the results showed that the frequencies of the constructed haplotypes of ecNOS, PC-1 and ACE genes were associated with CAD.Genotpyes of CRP,ACE and MTHFR genes were associated with serum CRP,ACE and Hcy level.(2) Confirmed by single plex PCR and capillary electrophoresis,the size of each gene's PCR production was consistent with original designed size.The specific of each target gene was confirmed according the capillary electrophoresis size of each gene's PCR production,control gene and size standard.The appropriate primer concentration of genes which were expressed abundant in peripheral blood white cells were confirmed by twofold serial dilutions in multiplexed PCR. In the current work we found the house keeping genes expression ofβ-actin and SRP14 were more stable than 18S-rRNA between the CAD and normal control groups,and the geometry average ofβ-actin and SRP14 could be regarded as normalization factor for quantification of target genes in the current work.We detected 14 genes expression profile in CAD patients and normal control subjects with the method described as above,and found expressions of ecNOS, SELL,ADD1 and MTHFR in the peripheral blood white cells were significantly different in both groups.The expression levels of these four genes were associated with genotypes of some SNPs.
[Conclusion]Multiplexed SNPs genotyping and genes expression methods established in the current work were quick,high-throughput,easy operation and microanalyse.Six SNPs and 4 genes discovered in the work that were associated with CAD may help in explaining the molecular bases of the disorder and the susceptibility to coronary atherosclerosis.It also can guide early predicting, preventing and therapy for CAD.
[方法](1)多重SNP基因分型的检测:提取外周血白细胞DNA,选择HapMap上公布的ADD1,PECAM-1,CRP,ecNOS,PC-1,SELL,GNB3,ACE,AT1R,AGT,ICAM-1,MTHFR和HL基因中国汉族人群的44个标签SNP,每个SNP设计3条引物(2条PCR扩增引物和1条延伸引物),通过PCR扩增、延伸及杂交,应用SNP stream基因分型系统对SNP位点进行基因分型。(2)多重基因表达水平的检测:提取外周血白细胞RNA,选择MTHFR,IL-8,ecNOS,TLR4,FOS,ID2,ADD1,ICAM-1,PECAM-1,CRP,LDLR,vWF,SELL,TNF-α等14个目标基因和β-actin、SRP14及18S-rRNA等3个管家基因,每个基因设计2条引物(1条反转录引物和1条PCR扩增引物),通过反转录和PCR扩增,应用高通量的GenomeLab GeXP分析仪检测17个基因的表达水平。(3)应用全自动生化分析仪检测血脂、CRP、ACE及Hcy水平。
[结果](1)构建的44个SNP分型成功32个,分型成功率为72.7%(32/44)。通过对583份DNA标本进行检测,32个SNP的平均样本检出率为98.3%。通过对中国汉族CAD患者和健康对照人群的32个SNP进行检测,结果显示CRP,ecNOS,tPc-1和ACE基因中的6个SNP的基因型频率和(或)等位基因频率在两组间存在统计学差异。根据连锁不平衡水平构建单倍型,结果显示ecNOS,PC-1,ACE基因的单倍型频率与CAD相关。实验发现CRP,ACE和MTHFR基因多态性分别与血清CRP,ACE和Hcy水平相关。(2)通过单重基因扩增、电泳,确认每个基因扩增产物的大小。结果显示扩增产物的大小与设计大小基本一致。根据每个基因扩增产物、质控基因和内标的片段大小,确认实验设计的每个基因扩增产物的特异性。在多重基因扩增中,根据电泳结果对外周血白细胞中高丰度表达基因的引物进行倍比稀释,确定了体系中每个基因的最佳引物浓度。通过对3个管家基因在两组之间表达水平的比较,发现以β-actin和SRP14基因的几何均数作为标准化因子可用以标准化本体系目标基因的表达。应用上述构建的14重基因表达检测方法对CAD患者和健康对照人群外周血白细胞RNA的基因表达水平进行检测,发现ecNOS,SELL,ADD1和MTHFR基因的表达水平在两组间存在统计学差异,其表达水平与部分SNP的基因型有关。
[结论]本实验构建的32重sNP基因分型和14重基因表达检测方法具有快速、准确、高通量、操作简单、微量化等优点。实验发现的与CAD相关的6个sNP位点和4个表达差异基因为心血管危险因素的预测和诊断提供了一种新思路,为心血管疾病寻找更有效的防治途径,早期预防和及时治疗提供理论指导。
[Objective]Coronary artery disease(CAD) is the leading cause of morbidity and mortality in recent years in China.It is a multifactorial disease caused by various genetic and environmental factors involved in accumulation of atherosclerosis in the walls of the coronary arteries.In order to find single nucleotide polymorphisms(SNP) and genes that were associated with CAD, multiplexed SNPs genotyping and genes expression methods were established in the current work.With the multiplexed methods,lots of tag SNPs and genes in peripheral blood white cells from CAD patients and normal control individuals were investigated.
[Methods](1) The method of multiplexed SNPs genotyping:DNA samples were extracted from peripheral blood white cells and 44 tag SNPs of 13 genes (ADD1,PECAM-1,CRP,ecNOS,PC-1,SELL,GNB3,ACE,AT1R,AGT, ICAM-1,MTHFR and HL) that were listed in the International Haplotype Map Project Database for CHB(Chinese Han nationality in Beijing) were selected in the current work.Three primers were designed for each SNP(two for polymerase chain reaction(PCR) and one for extension reaction).After PCR, extension and hybridization,genotypes of SNPs were investigated with the GenomeLab SNPstream genotyping system.(2) The method of multiplexed genes expression:RNA samples were extracted from peripheral blood white cells and two primers(one for reverse transcription and one for PCR) were designed for each of the 14 target genes(MTHFR,IL-8,ecNOS,TLR4,FOS, ID2,ADD1,ICAM-1,PECAM-1,CRP,LDLR,vWF,SELL and TNF-α) and 3 house keeping genes(β-actin,SRP14 and 18S-rRNA).After reverse transcription and PCR,expression levels of the 17 genes were detected with high-throughput GenomeLab GeXP Genetic Analysis System.(3) The levels of serum lipid,CRP,ACE and Hcy were detected with automatic biochemical analysis system.
[Result](1) Thirty two SNPs were genotyped and the rate of succeed SNP genotyping was 72.7%(32/44).The average genotyped rate of the 32 SNPs was 98.3%in the test of 583 DNA samples.With the 32 plexes SNPs genotyping method,DNA samples of CAD patients and normal control individuals from the local ethnic Han Chinese population were investigated.The results showed that the genotype and/or allele frequencies of 6 tag SNPs in CRP,ecNOS,PC-1 and ACE genes were significantly different in both groups.According the linkage-disequilibrium levels of SNPs,the haplotypes were constructed and the results showed that the frequencies of the constructed haplotypes of ecNOS, PC-1 and ACE genes were associated with CAD.Genotpyes of CRP,ACE and MTHFR genes were associated with serum CRP,ACE and Hcy level.(2) Confirmed by single plex PCR and capillary electrophoresis,the size of each gene's PCR production was consistent with original designed size.The specific of each target gene was confirmed according the capillary electrophoresis size of each gene's PCR production,control gene and size standard.The appropriate primer concentration of genes which were expressed abundant in peripheral blood white cells were confirmed by twofold serial dilutions in multiplexed PCR. In the current work we found the house keeping genes expression ofβ-actin and SRP14 were more stable than 18S-rRNA between the CAD and normal control groups,and the geometry average ofβ-actin and SRP14 could be regarded as normalization factor for quantification of target genes in the current work.We detected 14 genes expression profile in CAD patients and normal control subjects with the method described as above,and found expressions of ecNOS, SELL,ADD1 and MTHFR in the peripheral blood white cells were significantly different in both groups.The expression levels of these four genes were associated with genotypes of some SNPs.
[Conclusion]Multiplexed SNPs genotyping and genes expression methods established in the current work were quick,high-throughput,easy operation and microanalyse.Six SNPs and 4 genes discovered in the work that were associated with CAD may help in explaining the molecular bases of the disorder and the susceptibility to coronary atherosclerosis.It also can guide early predicting, preventing and therapy for CAD.
引文
[1]Wilsom P W.Assessing coronary heart disease risk with traditional and novel risk factors[J].Clin Cardiol,2004,27(6Supp13):7-11.
[2]Ma J,Dempsy A,Stamatiou D,et al.Identifying leukocyte gene expression patterns associated with plasma lipid levels in human subjects[J].Atherosclerosis,2007,191(1):63-72.
[3]Vieman D,Barczyk K,Vogi T,et al.MRP8/MRP14 impairs endothelial integrity and induces a caspase-dependent and independent cell death program[J].Blood,2007,109(6):2453-2460.
[4]Li J J,Fang CH.C-reactive protein is not only an inflammatory marker but also a direct cause of cardiovascular diseases[J].Med Hypotheses,2004,62(4):499-506.
[5]Alanna C.M,Molly S.B,Aaron R.F,et al.ADD1 460W allele associated with cardiovascular disease in hypertensive individuals[J].Hypertension,2002,39(6):1053-1057.
[6]黄晓红,孙凯,宋燕等。α-内收蛋白基因及G蛋白β3亚单位基因多态与原发性高血压的关系[J]。中华医学杂志,2007,87(24):1682-1684。
[7]王丛,孙刚,丁燕程等。α-内收蛋白基因多态性与大动脉弹性相关性研究[J]。中国分子心脏病学杂志,2007,7(4):214-217。
[8]Yan L,Lutgarde T,Tatiana K,et al.Cardiovascular risk in relation to α-Adducin Gly460Trp polymorphism and systolic pressure[J].Hypertension,2005,46(3):527-532.
[9]潘闽,蒋文平,刘志华等。PECAM-1单核苷酸多态性与中国人群冠心病的相关性[J]。江苏医药,2004,34(4):366-367。
[10]常智堂,陈健,程龙献等。血小板内皮细胞黏附分子-1基因Leu 125 Val 多态性与冠心病及危险因素的关系[J]。中华老年心脑血管病杂志,2007,9(10):664-667。
[11]Lu Fang,Heming Wei,Sanual H.C,et al.Association of Leu125Val polymorphism of platelet endothelial cell adhesion molecule-1(PECAM-1)gene & soluble level of PECAM-1 with coronary artery disease in Asian Indians[J].Indian J Med Res,2005,121(2):92-99.
[12]Wei H,Fang L,Chowdhury SH,et al.Platelet-endothelial cell adhesion molecule-1 gene polymorphism and its soluble level are associated with severe coronary artery steecNOSis in Chinese Singaporean[J].Clin Biochem,2004,37(12):1091-1097.
[13]Rifai N,Ridker PM.High-sensitivity C-reactive Protein:a novel and promising marker of coronary heart disease[J].Clin Chem,2001,47(3):403-411.
[14]Kinjo K,Sato H,Ohnishi Y,et al.Impact of high-sensitivity C-reactive protein on predicting long-term mortality of acute myocardial infarction[J].Am J Cardiol,2003,91(8):931-935.
[15]Roy D,Quiles J,Avanzas P,et al.A comparative study of markers of inflammation for the assessment of cardiovascular risk in patients presenting to the emergency department with acute chest pain suggestive of acute coronary syndrome[J].Int J Cardiol,2006,109(3):317-321.
[16]Jahn J,Hellmann I,Maass M,et al.Time dependent changes ofhs-CRP serum concentration in patients with non ST elevation acute coronary syndrome[J].Herz,2004,29(8):795-801.
[17]Yip HK,Hung WC,Yang CH,et al.Serum concentrations of high-sensitivity C-reactive protein predict progressively obstructive lesions rather than late resteecNOSis in patients with unstable angina undergoing coronary artery stenting[J].Circ J,2005,69(10):1202-1207.
[18]丁艳萍,马周建。C反应蛋白及其基因多态性与冠心病发病的相关性研究[J]。山东医药,2006,46(19):36-37。
[19]夏晖,刘厂辉,阳辉等。内皮型一氧化氮合酶基因G894T突变及低镁血症与中国汉族冠心病的相关性[J]。南华大学学报,2007,35(5):664-667。
[20]Rossi GP,Cesari M,Zanchetta M,et al.The T-786C endothelial nitric oxide synthase genotype is a novel risk factor for coronary artery disease in Caucasian patients of the GENICA study[J].Am Coll Cardiol,2003,41-(6):930-937.
[21]Tangurek B,Ozer N,Sayar N,et al.The relationship between endothelial nitric oxide synthase gene polymorphism(T-786 C) and coronary artery disease in the Turkish population[J].Heart Vessels,2006,21(5):285-290.
[22]Lin NT,Lee M J,Lee RP,et al.Analysis of endothelial nitric oxide synthase gene polymorphisms with cardiovascular diseases in eastern Taiwan[J].Chin J Physiol,2008,51(1):42-47.
[23]于辉,郑红,赵洛沙等。河南汉族冠心病患者内皮型一氧化氮合酶基因多态性分析[J]。郑州大学学报,2006,41(2):293-295。
[24]冉军川,白锋,张钲等。eNOS基因5'侧翼区T-786 C多态性与冠心病的相关性研究[J]。临床内科杂志,2006,23(10):689-691。
[25]李梅蕊,吴于滨,刘华等。PC-1基因多态性与2型糖尿病及糖尿病肾病的相关性研究[J]。昆明医学院学报,2008,29(1):27-31。
[26]Simonetta B,Ornella L,Sabrina P,et al.The K121Q polymorphism of the ENPP1/PC-1 gene is associated with insulin resistance/atherogenic phenotypes,including earlier onset of type 2 diabetes and myocardial infarction[J].Diabetes,2005,54(10):3021-3025.
[27]陈肖俊,汪大望,吴建波等。G蛋白β3亚单位C825 T等位基因多态性与原发性高血压的关系[J]。临床内科杂志,2007,24(5):333-334。
[28]Von B,Schusterschitz Y,Koch W,et al.G protein beta 3 subunit 825T allele carriage and risk of coronary artery disease[J].Atherosclerosis,2003,167(1):135- 139.
[29]夏尊恩,李艳,熊小泉等。老年冠心病患者L-选择素P213S基因多态性研究[J]。中国老年学杂志,2006,26(5):594-596。
[30]Gardemann A,Weiss T,Schwartz O,et al.Gene polymorphism but not catalytic activity of angiotensin I-converting enzyme is associated with coronary artery disease and myocardial infarction in low-risk patients[J]. Circulation,1995,92(10):2796-2799.
[31]Acarturk E,Attila G,Bozkurt A,et al.Insertion/deletion polymorphism of the angiotensin converting enzyme gene in coronary artery disease in southern Turkey[J].J Biochem Mol Biol,2005,38(4):486-490.
[32]Zak I,Niemiec P,Sarecka B,et al.Cartier-state of D allele in ACE gene insertion/deletion polymorphism is associated with coronary artery disease,in contrast to the C677T transition in the MTHFR gene[J].Acta Biochim Pol,2003;50(2):527-534.
[33]张奎星,刘同宝,徐秋霞等。血管紧张素Ⅱ 1型受体基因单核苷酸多态性与原发性高血压和冠心病的相关研究[J]。中华心血管病杂志,2005,33(8):720-723。
[34]徐欣,徐雯,杨承健等。血管紧张素Ⅱ-1型受体基因A1 166/C多态性与中国人冠心病的关系[J]。山东医药,2007,47(27):135-136。
[35]杨涛,石毅,陈敬州等。血管紧张素Ⅱ 1型受体基因多态性与冠心病发病相关[J]。中国分子心脏病学杂志,2008年,8(2):91-94。
[36]杨帆,赵洛沙,郑红等。血管紧张素原基因G-6A多态性与冠心病的相关性研究[J]。山东医药,2008,48(1):19-21。
[37]任洁,贾永平,吕吉元等。血管紧张素原基因M235T多态性与冠心病的相关性研究[J]。山西医科大学学报,2005,36(4):439-442。
[38]Robert Y.L.Z,Suzanne C,Henry A.E,et al.Intercellular adhesion molecule 1(ICAM1) Lys56Met and Gly241Arg gene variants,plasma-soluble ICAM1concentrations,and risk of incident cardiovascular events in 23 014 Initially healthy white women[J].Stroke,2007,38(12):3152-3157.
[39]周有利,朱名安,丁妍等。细胞间黏附分子-1基因多态性与冠心病相关性研究[J]。临床心血管病杂志,2006,22(9):519-522。
[40]韦叶生,唐任光,袁锡华等。细胞间黏附分子-1基因K469E多态性与冠心病关系的研究[J]。中国免疫学杂志,2006,22(11):1056-1059。
[41]汪明,李艳,张平安等。冠心病患者细胞问黏附分子1基因多态性与其他暴露因素的交互作用的研究[J]。中华检验医学杂志,2006,29(12): 1123-1128.
[42]Mager A,Koren-Morag N,Shohat M,et al.Impact of ethnicity and MTHFR genotype on age at onset of coronary artery disease in women in Israel[J].Isr Med Assoc J,2008,10(7):516-519.
[43]Jos(?) M.G P,Salvador E C,Manuel J N,et al.Influence of 677 C→T polymorphism of methylenetetrahydrofolate reductase on medium-term progecNOSis after acute coronary syndromes[J].Tex Heart Inst J,2007,34(2):142-147.
[44]潘棱,沈晓丽,林立芳等。MTHFR基因多态性及血浆同型半胱氨酸水平与冠心病的关系[J]。心脏杂志,2006,18(5):542-543。
[45]祝鸿喜,马向红,黄体钢等。MTHFR基因多态性与冠心病的关系[J]。实用心脑肺血管病杂志,2007,15(8):569-572。
[46]Ghazouani L,Abboud N,Mtiraoui N,et al.Homocysteine and methylenetetrahydrofolate reductase C677T and A1298C polymorphisms in Tunisian patients with severe coronary artery disease[J].J Thromb Thrombolysis,2009,27(2):191-197.
[47]Laraqui A,Allami A,Carri(?) A,et al.Relation between plasma homocysteine,gene polymorphisms ofhomocysteine metabolism-related enzymes,and angiographically proven coronary artery disease[J].Eur J Intern Med,2007,18(6):474-483.
[48]Kerkeni M,Addad F,Chauffert M,et al.Hyperhomocysteinaemia,methylenetetrahydrofolate reductase polymorphism and risk of coronary artery disease[J].Ann Clin Biochem,2006,43(Pt 3):200-206.
[49]倪培华,应雅韵,傅毅等。肝脂酶基因启动子多态性与连锁不平衡分析[J]。诊断学理论与实践,2007,6(4):339-342。
[50]施金俏,倪培华,郑寿贵等。肝脂酶基因启动子250G/A多态性与冠心病的关系[J]。中华检验医学杂志,2006,29(6):515-517。
[51]胡敏,邵建国,朱毅等。肝脂肪酶基因-514C/T多态性与冠心病相关性研究[J]。中国药理学通报,2006,22(9):1118-1121。
[52]姜红,杜明,西雁等。Fc Y受体Ⅱ A基因多态性与冠心病和扩张型心肌病的关系[J]。国际遗传学杂志,2006,29(4):246-249。
[53]梁茜,董吁钢,杨希立等。基因芯片技术分析老年冠心病患者的易感基因[J]。中华老年医学杂志,2006,25(6):520-523。
[54]Pereira AC,Miyakawa AA,Lopes NH,et al.Dynamic regulation of MTHFR mRNA expression and C677T genotype modulate mortality in coronary artery disease patients after revascularization[J].Thromb Res,2007;121(1):25-32.
[55]Satterthwaite G,Francis SE,Suvarna K,et al.Differential gene expression in coronary arteries from patients presenting with ischemic heart disease:further evidence for the inflammatory basis of atherosclerosis[J].Am Heart J,2005,150(3):488-499.
[56]O'Malley T,Ludlam C.A,Riemermsa R.A,et al.Early increase in levels of soluble inter-cellular adhesion molecule-1(sICAM-1)[J].European Heart Journal,2001(22),1226-1234.
[57]Atalar E,Aytemir K,Haznedaro(?)lu I,et al.Increased plasma levels of soluble selectins in patients with unstable angina[J].Int J Cardiol,2001,78(1):69-73.
[58]Inoue T,Komoda H,Nonaka M,et al.Interleukin-8 as an independent predictor of long-term clinical outcome in patients with coronary artery disease[J].Int J Cardiol,2008,124(3):319-325.
[59]Dominguez-Rodriguez A,Abreu-Gonzalez P,Garcia-Gonzalez M,et al.Prognostic value of interleukin-8 as a predictor of heart failure in patients with myocardial infarction and percutaneous intervention[J].Int J Cardiol,2006,111(1):158-160.
[60]Xiaoyong QI,Shuren LI,Junyong LI,et al.The prognostic value of IL-8 for cardiac events and restenosis in patients with coronary heart diseasesafter percutaneous coronary intervention[J].Jpn Heart J,2003,44(5):623-672.
[61]Xiaoyong QI,Junyong LI,Jian GU,et al.Plasma levels of IL-8 predict early complications in patients with coronary heart disease after percutaneous coronary intervention[J].Jpn Heart J,2003,44(4):451-461.
[62]Shimasaki Y,Saito Y,Yoshimura M,et al.The effects of long-term smoking on endothelial nitric oxide synthase mRNA expression in human platelets as detected with real-time quantitative RT-PCR[J].Clin Appl Thromb Hemost,2007,13(1):43-51.
[63]Shiraki R,Inoue N,Kobayashi S,et al.Toll-like receptor 4 expressions on peripheral blood monocytes were enhanced in coronary artery disease even in patients with low C-reactive protein[J].Life Sci,2006,80(1):59-66.
[64]Geng HL,Lu HQ,Zhang LZ,et al.Increased expression of Toll like receptor 4 on peripheral-blood mononuclear cells in patients with coronary arteriosclerosis disease[J].Clin Exp Immunol,2006,143(2):269-273.
[65]Willmar D,Patino,Omar Y,et al.Circulating transcriptome reveals markers of atherosclerosis[J].Proc Natl Acad Sci USA,2005,102(9):3423-3428.
[66]Anothai P,Roger R.T,Ian J,et al.LDL-Receptor mRNA expression in men is downregulated within an hour of an acute fat load and is influenced by genetic polymorphism[J].J Nutr,2007,137(9):2062-2067.
[67]Wang IJ,Chiang TH,Shih YF,et al.The association of single nucleotide polymorphisms in the MMP-9 genes with susceptibility to acute primary angle closure glaucoma in Taiwanese patients[J].Molecular Vision,2006,26(12):1223-1232.
[68]Pollin TI,Tanner K,Connell JR,et al.Linkage of plasma adiponectin levels to 3q27 explained by association with variation in the APM1 gene[J].Diabetes,2005,54(1):268-274.
[69]Damcott CM,Ott SH,Pollin TI,et al.Genetic variation in adiponectin receptor 1 and adiponectin receptor 2 is associated with type 2 diabetes in the old order Amish[J].Diabetes,2005,54(7):2245-2250.
[70]Akihiko M,Tomohiro N,Nobutaka D,et al.Polymorphism of the C-reactive protein (CRP) gene is related to serum CRP level and arterial pulse wave velocity in healthy elderly Japanese[J].Hypertens Res,2006,29(5):323-331.
[71]Russell Al,Cunninghame G DS,Shepherd C,et al.Polymorphism at the C-reactive protein locus influences gene expression and predisposes to-systemic lupus erythematosus[J].Hum Mol Genet,2004,13(1):137-147.
[72]Brull DJ,Serrano N,Zito F,et al.Human CRP gene polymorphism influences CRP levels:implications for the prediction and pathogenesis of coronary heart disease[J].Arterioscler Thromb Vasc Biol,2003,23(11):2063-2069.
[73]Zee RY,Ridker PM.Polymorphism in the human C-reactive protein (CRP) gene,plasma concentrations of CRP,and the risk of future arterial thrombosis[J].Atherosclerosis,2002,162(1):217-219.
[74]Szalai AJ,McCrory MA,Cooper GS,et al.Association between baseline levels of C-reactive protein (CRP) and a dinucleotide repeat polymorphism in the intron of the CRP gene[J].Genes Immun,2002,3(1):14-19.
[75]Dana C.C,Christopher L.S,Xiaoting Qin,et al.Genetic variation is associated with C-reactive protein levels in the third national health and nutrition examination survey[J].Circulation,2006,114 (23):2458-2465.
[76]Morita A,Nakayama T,Soma M.Association study between C-reactive protein genes and ischemic stroke in Japanese subjects[J].Am J Hypertens,2006,19 (6):593-600.
[77]Shen J,Arnett DK,Parnell LD,et al.Association of common C-reactive protein (CRP) gene polymorphisms with baseline plasma CRP levels and fenofibrate response:The GOLDN Study[J].Diabetes Care,2008,31(5):910-915.
[78]Kolz M,Koenig W,Müller M,et al.DNA variants,plasma levels and variability of C-reactive protein in myocardial infarction survivors:results from the AIRGENE study[J].Eur Heart J,2008,29(10):1250-1258.
[79]Mart(?)nez-Calatrava MJ,Gonz(?)lez-S(?)nchez JL,Mart(?)nez-Larrad MT,et al.Common haplotypes of the C-reactive protein gene and circulating leptin levels influence the interindividual variability in serum C-reactive protein levels[J].Thromb Haemost,2007,98(5):1088-1095.
[80]Wang XL,Mahaney MC,Sim AS,et al.Genetic contribution of the endothelial constitutive nitric oxide synthase gene to plasma nitric oxide levels[J].Arterioscler Thromb Vase Biol,1997,17(11):3147-3153.
[81]Tasic I,Milojkovic M,Sunder-Plassmann R,et al.The association of PC-1(ENPP1) K121Q polymorphism with metabolic syndrome in patients with coronary heart disease[J].Clin Chim Acta,2007,377(1-2):237-242.
[82]Hyun-Ju Seo,Soo-Geun Kim,Oh-Joong Kwon.The K121Q polymorphism in ENPP1(PC-1) is not associated with type 2 diabetes or obesity in Korean Male workers[J].J Korean Med Sci,2008,23(3):459-464.
[83]Jacek B,Grzegorz P,Krzysztof W,et al.New polymorphism of ENPP1(PC-1) is associated with increased risk of type 2 diabetes among obese individuals[J].Diabetes,2006,55(9):2626-2630.
[84]Kretowski A,McFann K,Hokanson JE,et al.Polymorphisms of the renin-angiotensin system genes predict progression of subclinical coronary atherosclerosis[J].Diabetes,2007,56(3):863-871.
[85]Guneri S,Baris N,Aytekin D,et al.The relationship between angiotensin converting enzyme gene polymorphism,coronary artery disease,and stent resteecNOSis[J].Int Heart J,2005,46(5):889-897.
[86]卢杨,屈会起。中国心肌梗死患者血管紧张素I转换酶基因插入/缺失多态性Meta分析[J]。循证医学,2001,1(1):18-21。
[87]李丽华,王曼丽,赵红等。冠心病患者血管紧张素转换酶基因多态性分析[J]。山东医药,2008,48(2):41-42。
[88]Stavroulakis GA,Makris TK,Krespi OG,et al.Predicting respose to chronic antihypertensivetreatment with fosinopril:the role of angiotensin converting enzyme gene polymorphism[J].Cardiovasc Drugs Ther,2000,14(4):427-432.
[89]L Tiret,B Rigat,S Visvikis,C Breda,et al.Evidence,from combined segregation and linkage analysis,that a variant of the angiotensin I-converting enzyme(ACE) gene controls plasma ACE levels[J].Am J Hum Genet,1992,51(1):197-205.
[90]Gordon V,Patrick P,Robert S,et al.Gene expression analysis of troglitazone reveals its impact on multiple pathways in cell culture:a case for in vitro platforms combined with gene expression analysis for early(idiosyncratic)toxicity screening[J].Inter J Toxicology,2006,25(2):85-94.
[91]Chen Q R,Gordon V,Kahuku O,et al.Diagnosis of the small round blue cell tumors using multiplex polymerase chain reaction[J].J Molecular Diagnostics,2007,9(1):80-88.
[02]Jean D.B,Tara M.R,Michele A.K,et al.Protection against simian/human immunodeficiency virus(SHIV) 89.6P in macaques after coimmunization with SHIV antigen and IL-15 plasmid[J].Proc Natl Acad Sci,2007,104(47):18648-18653.
[93]Vandesompele J,De P K,Pattyn F,et al.Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes[J].Genome Biol,2002,3(7):RESEARCH0034.
[94]Huggett J,Dheda K,Bustin S,et al.Real-time RT-PCR normalisation;strategies and considerations[J].Genes Immun,2005,6(4):279- 284.
[95]Tsuji N,Kamagata C,Furuya M,et al.Selection of an internal control gene for quantitation of mRNA in colonic tissues[J].Anticancer Res,2002,22(6C):4173-4178.
[96]Fuster G,Vicente R,Coma M,et al.One-step reverse transcription polymerase chain reaction for semiquantitative analysis of mRNA expression[J].Methods Find Exp Clin Pharmacol,2002,24(5):253-259.
[97]Ajay P S A,Sarvesh R,Sachinandan De,et al.Expression stability of two housekeeping genes(18S rRNA and G3PDH) during in vitro maturation of follicular oocytes in buffalo(Bubalus bubalis)[J].Animal Reproduction Science,2008,103(1-2):164-171.
[98]陈应泰,姜冠潮,王丹等。18srRNA作为内部参考在肺癌相关基因LSCC-3研究中的应用[J]。广西医科大学学报,2005,22(2):209-211。
[99]Pilbrow AP,Ellmers LJ,Black MA,et al.Genomic selection of reference genes for real-time PCR in human myocardium[J].BMC Med Genomics,2008,29;1 (1):64.
[100]Pierre V,Marc R,Tanveer A.K,et al.Differences in gene expression profiles of diabetic and nondiabetic patients undergoing cardiopulmonary bypass and cardioplegic arrest[J].Circulation,2004,110(2):280-286.
[101]Hendrik J.M.de J,Rudolf S.N.F,Eveline S.J.M.de B,et al.Evidence based selection of housekeeping genes[J].PLoS ONE,2007,2(9):e898.
[102]Teupser D,Mueller MA,Koglin J,et al.CD36 mRNA expression is increased in CD 14+ monocytes of patients with coronary heart disease[J].Clin Exp Pharmacol Physiol,2008,35(5-6):552-556.
[103]Scherrer-Crosbie M,UUrich R,Bloch KD,et al.Endothelial nitric oxide synthase limits left ventricular remodeling after myocardial infarction in mice[J].Circulation,2001,104(11):1286-1291.
[104]Jones SP,Greer JJ,van Haperen R,et al.Endothelial nitric oxide synthase overexpression attenuates congestive heart failure in mice[J].Proc Natl Acad Sci,2003,100(8):4891-4896.
[105]Akira K,Shin M,Takahiro M,et al.Increased expression of monocyte CD11a and intracellular adhesion molecule-1 in patients with initial atherosclerotic coronary stenosis[J].Circ J,2004;68(1):6-10.
[106]Bowen SL,Bloor BK,Leigh IM,et al.Adducin expression in cutaneous and oral lesions:alpha-and beta-adducin transcripts down-regulate with keratinocyte differentiation in stratified epithelia[J].J Pathol,2003,201 (1):119-126.
[107]Huijun S,Tomonari K,Tomonaga I,et al.C-reactive protein in atherosclerotic lesions[J].Am J Pathol.2005,167(4):1139-1148.
[108]Randall S,Sherri O L,Lisa H,et al.C-reactive protein downregulates endothelial NO synthase and attenuates reendothelialization in vivo in mice[J].Circulation Research,2007;100(10):1452-1459.
[109]Senthil K V,Sridevi D,Ivan Y,et al.Demonstration that C-reactive protein decreases eNOS expression and bioactivity in human aortic endothelial cells[J].Circulation,2002;106(12):1439-1441.
[110]Paolo C,James T.W,Edward T.H.Y,et al.Inflammatory cytokines stimulated C-reactive protein production by human coronary artery smooth muscle cells[J].Circulation,2003;108(16):1930-1932.
[1]Hristov M,Erl W,Weber PC.Endothelial progenitor cells:mobilization,differentiation,and homing.Arterioscler Thromb Vasc Biol,2003,23(7):1185-1189.
[2]Werner N,Kosiol S,Schiegl,et al.Circulating endothelial progenitor cells and cardiovascular outcomes.N Engl J Med,2005,353(10):999- 1007.
[3]Schmidt-Lucke C,Rossig L,Fichtlscherer S,et al.Reduced number of circulating endothelial progenitor cells predicts future cardiovascular events:proof of concept for the clinical importance of endogenous vascular repair.Circulation,2005,111(22):2981-2987.
[4]Blann AD,Woywodt A,Bertolini F,et al.Circulating endothelial cells.Biomarker of vascular disease.Thromb Haemost,2005,93(2):228-235.
[5]Massa M,Rosti V,Ferrario M,et al.Increased circulating hematopoietic and endothelial progenitor cells in the early phase of acute myocardial infarction.Blood,2005,105(1):199-206.
[6]George J,Goldstein E,Abashidze S,et al.Circulating endothelial progenitor cells in patients with unstable angina:association with systemic inflammation.Eur Heart J,2004,25(12):1003-1008.
[7]Adams V,Lenk K,Linke A,et al.Increase of circulating endothelial progenitor cells in patients with coronary artery disease after exercise-induced ischemia.Arterioscler Thromb Vasc Biol,2004,24(4):684-690.
[8]Fadini GP,Sartore S,Albiero M,et al.Number and function of endothelial progenitor cells as a marker of severity for diabetic vasculopathy.Arterioscler Thromb Vasc Biol,2006,26(9):2140-2146.
[9]Pirro M,Schillaci G,Paltriccia R,et al.Increased ratio of CD31+/CD42-microparticles to endothelial progenitors as a novel marker of atherosclerosis in hypercholesterolemia.Arterioscler Thromb Vasc Biol,2006,26(11):2530-2535.
[10]HenrichD,HahnP,WahlM,etal.Serum derived from multiple trauma patients promotes the differentiation of endothelial progenitor cells in vitro:possible role of transforming growth factor-betal and vascular endothelial growth factor165.Shock,2004,21(1):13-16.
[11]Kunz GA,Liang G,Cuculi F,et al.Circulating endothelial progenitor cells predict coronary artery disease severity.Am Heart J,2006,152 (1):190-195.
[12]Laufs U,Werner N,Link A,et al.Physical training increases endothelial progenitor cells,inhibits neointima formation,and enhances angiogenesis.Circulation,2004,109(2):220-226.
[13]Schomig K,Busch G,Steppich B,et al.Interleukin-8 is associated with circulating CD 133+ progenitor cells in acute myocardial infarction.Eur Heart J,2006,27(9):1032-1037.
[14]YoonYS,Johnson LA,ParkJS,et al.Therapeutic myocardial angiogenesis with vascular endothelial growth factors.Mol Cell Biochem,2004,264 (1-2):63-74.
[15]Kusuyama T,Omura T,Nishiya D,et al.The effects of HMG-CoA reductase inhibitor on vascular progenitor cells.J Pharmacol Sci,2006,101 (4):344-349.
[2]Ma J,Dempsy A,Stamatiou D,et al.Identifying leukocyte gene expression patterns associated with plasma lipid levels in human subjects[J].Atherosclerosis,2007,191(1):63-72.
[3]Vieman D,Barczyk K,Vogi T,et al.MRP8/MRP14 impairs endothelial integrity and induces a caspase-dependent and independent cell death program[J].Blood,2007,109(6):2453-2460.
[4]Li J J,Fang CH.C-reactive protein is not only an inflammatory marker but also a direct cause of cardiovascular diseases[J].Med Hypotheses,2004,62(4):499-506.
[5]Alanna C.M,Molly S.B,Aaron R.F,et al.ADD1 460W allele associated with cardiovascular disease in hypertensive individuals[J].Hypertension,2002,39(6):1053-1057.
[6]黄晓红,孙凯,宋燕等。α-内收蛋白基因及G蛋白β3亚单位基因多态与原发性高血压的关系[J]。中华医学杂志,2007,87(24):1682-1684。
[7]王丛,孙刚,丁燕程等。α-内收蛋白基因多态性与大动脉弹性相关性研究[J]。中国分子心脏病学杂志,2007,7(4):214-217。
[8]Yan L,Lutgarde T,Tatiana K,et al.Cardiovascular risk in relation to α-Adducin Gly460Trp polymorphism and systolic pressure[J].Hypertension,2005,46(3):527-532.
[9]潘闽,蒋文平,刘志华等。PECAM-1单核苷酸多态性与中国人群冠心病的相关性[J]。江苏医药,2004,34(4):366-367。
[10]常智堂,陈健,程龙献等。血小板内皮细胞黏附分子-1基因Leu 125 Val 多态性与冠心病及危险因素的关系[J]。中华老年心脑血管病杂志,2007,9(10):664-667。
[11]Lu Fang,Heming Wei,Sanual H.C,et al.Association of Leu125Val polymorphism of platelet endothelial cell adhesion molecule-1(PECAM-1)gene & soluble level of PECAM-1 with coronary artery disease in Asian Indians[J].Indian J Med Res,2005,121(2):92-99.
[12]Wei H,Fang L,Chowdhury SH,et al.Platelet-endothelial cell adhesion molecule-1 gene polymorphism and its soluble level are associated with severe coronary artery steecNOSis in Chinese Singaporean[J].Clin Biochem,2004,37(12):1091-1097.
[13]Rifai N,Ridker PM.High-sensitivity C-reactive Protein:a novel and promising marker of coronary heart disease[J].Clin Chem,2001,47(3):403-411.
[14]Kinjo K,Sato H,Ohnishi Y,et al.Impact of high-sensitivity C-reactive protein on predicting long-term mortality of acute myocardial infarction[J].Am J Cardiol,2003,91(8):931-935.
[15]Roy D,Quiles J,Avanzas P,et al.A comparative study of markers of inflammation for the assessment of cardiovascular risk in patients presenting to the emergency department with acute chest pain suggestive of acute coronary syndrome[J].Int J Cardiol,2006,109(3):317-321.
[16]Jahn J,Hellmann I,Maass M,et al.Time dependent changes ofhs-CRP serum concentration in patients with non ST elevation acute coronary syndrome[J].Herz,2004,29(8):795-801.
[17]Yip HK,Hung WC,Yang CH,et al.Serum concentrations of high-sensitivity C-reactive protein predict progressively obstructive lesions rather than late resteecNOSis in patients with unstable angina undergoing coronary artery stenting[J].Circ J,2005,69(10):1202-1207.
[18]丁艳萍,马周建。C反应蛋白及其基因多态性与冠心病发病的相关性研究[J]。山东医药,2006,46(19):36-37。
[19]夏晖,刘厂辉,阳辉等。内皮型一氧化氮合酶基因G894T突变及低镁血症与中国汉族冠心病的相关性[J]。南华大学学报,2007,35(5):664-667。
[20]Rossi GP,Cesari M,Zanchetta M,et al.The T-786C endothelial nitric oxide synthase genotype is a novel risk factor for coronary artery disease in Caucasian patients of the GENICA study[J].Am Coll Cardiol,2003,41-(6):930-937.
[21]Tangurek B,Ozer N,Sayar N,et al.The relationship between endothelial nitric oxide synthase gene polymorphism(T-786 C) and coronary artery disease in the Turkish population[J].Heart Vessels,2006,21(5):285-290.
[22]Lin NT,Lee M J,Lee RP,et al.Analysis of endothelial nitric oxide synthase gene polymorphisms with cardiovascular diseases in eastern Taiwan[J].Chin J Physiol,2008,51(1):42-47.
[23]于辉,郑红,赵洛沙等。河南汉族冠心病患者内皮型一氧化氮合酶基因多态性分析[J]。郑州大学学报,2006,41(2):293-295。
[24]冉军川,白锋,张钲等。eNOS基因5'侧翼区T-786 C多态性与冠心病的相关性研究[J]。临床内科杂志,2006,23(10):689-691。
[25]李梅蕊,吴于滨,刘华等。PC-1基因多态性与2型糖尿病及糖尿病肾病的相关性研究[J]。昆明医学院学报,2008,29(1):27-31。
[26]Simonetta B,Ornella L,Sabrina P,et al.The K121Q polymorphism of the ENPP1/PC-1 gene is associated with insulin resistance/atherogenic phenotypes,including earlier onset of type 2 diabetes and myocardial infarction[J].Diabetes,2005,54(10):3021-3025.
[27]陈肖俊,汪大望,吴建波等。G蛋白β3亚单位C825 T等位基因多态性与原发性高血压的关系[J]。临床内科杂志,2007,24(5):333-334。
[28]Von B,Schusterschitz Y,Koch W,et al.G protein beta 3 subunit 825T allele carriage and risk of coronary artery disease[J].Atherosclerosis,2003,167(1):135- 139.
[29]夏尊恩,李艳,熊小泉等。老年冠心病患者L-选择素P213S基因多态性研究[J]。中国老年学杂志,2006,26(5):594-596。
[30]Gardemann A,Weiss T,Schwartz O,et al.Gene polymorphism but not catalytic activity of angiotensin I-converting enzyme is associated with coronary artery disease and myocardial infarction in low-risk patients[J]. Circulation,1995,92(10):2796-2799.
[31]Acarturk E,Attila G,Bozkurt A,et al.Insertion/deletion polymorphism of the angiotensin converting enzyme gene in coronary artery disease in southern Turkey[J].J Biochem Mol Biol,2005,38(4):486-490.
[32]Zak I,Niemiec P,Sarecka B,et al.Cartier-state of D allele in ACE gene insertion/deletion polymorphism is associated with coronary artery disease,in contrast to the C677T transition in the MTHFR gene[J].Acta Biochim Pol,2003;50(2):527-534.
[33]张奎星,刘同宝,徐秋霞等。血管紧张素Ⅱ 1型受体基因单核苷酸多态性与原发性高血压和冠心病的相关研究[J]。中华心血管病杂志,2005,33(8):720-723。
[34]徐欣,徐雯,杨承健等。血管紧张素Ⅱ-1型受体基因A1 166/C多态性与中国人冠心病的关系[J]。山东医药,2007,47(27):135-136。
[35]杨涛,石毅,陈敬州等。血管紧张素Ⅱ 1型受体基因多态性与冠心病发病相关[J]。中国分子心脏病学杂志,2008年,8(2):91-94。
[36]杨帆,赵洛沙,郑红等。血管紧张素原基因G-6A多态性与冠心病的相关性研究[J]。山东医药,2008,48(1):19-21。
[37]任洁,贾永平,吕吉元等。血管紧张素原基因M235T多态性与冠心病的相关性研究[J]。山西医科大学学报,2005,36(4):439-442。
[38]Robert Y.L.Z,Suzanne C,Henry A.E,et al.Intercellular adhesion molecule 1(ICAM1) Lys56Met and Gly241Arg gene variants,plasma-soluble ICAM1concentrations,and risk of incident cardiovascular events in 23 014 Initially healthy white women[J].Stroke,2007,38(12):3152-3157.
[39]周有利,朱名安,丁妍等。细胞间黏附分子-1基因多态性与冠心病相关性研究[J]。临床心血管病杂志,2006,22(9):519-522。
[40]韦叶生,唐任光,袁锡华等。细胞间黏附分子-1基因K469E多态性与冠心病关系的研究[J]。中国免疫学杂志,2006,22(11):1056-1059。
[41]汪明,李艳,张平安等。冠心病患者细胞问黏附分子1基因多态性与其他暴露因素的交互作用的研究[J]。中华检验医学杂志,2006,29(12): 1123-1128.
[42]Mager A,Koren-Morag N,Shohat M,et al.Impact of ethnicity and MTHFR genotype on age at onset of coronary artery disease in women in Israel[J].Isr Med Assoc J,2008,10(7):516-519.
[43]Jos(?) M.G P,Salvador E C,Manuel J N,et al.Influence of 677 C→T polymorphism of methylenetetrahydrofolate reductase on medium-term progecNOSis after acute coronary syndromes[J].Tex Heart Inst J,2007,34(2):142-147.
[44]潘棱,沈晓丽,林立芳等。MTHFR基因多态性及血浆同型半胱氨酸水平与冠心病的关系[J]。心脏杂志,2006,18(5):542-543。
[45]祝鸿喜,马向红,黄体钢等。MTHFR基因多态性与冠心病的关系[J]。实用心脑肺血管病杂志,2007,15(8):569-572。
[46]Ghazouani L,Abboud N,Mtiraoui N,et al.Homocysteine and methylenetetrahydrofolate reductase C677T and A1298C polymorphisms in Tunisian patients with severe coronary artery disease[J].J Thromb Thrombolysis,2009,27(2):191-197.
[47]Laraqui A,Allami A,Carri(?) A,et al.Relation between plasma homocysteine,gene polymorphisms ofhomocysteine metabolism-related enzymes,and angiographically proven coronary artery disease[J].Eur J Intern Med,2007,18(6):474-483.
[48]Kerkeni M,Addad F,Chauffert M,et al.Hyperhomocysteinaemia,methylenetetrahydrofolate reductase polymorphism and risk of coronary artery disease[J].Ann Clin Biochem,2006,43(Pt 3):200-206.
[49]倪培华,应雅韵,傅毅等。肝脂酶基因启动子多态性与连锁不平衡分析[J]。诊断学理论与实践,2007,6(4):339-342。
[50]施金俏,倪培华,郑寿贵等。肝脂酶基因启动子250G/A多态性与冠心病的关系[J]。中华检验医学杂志,2006,29(6):515-517。
[51]胡敏,邵建国,朱毅等。肝脂肪酶基因-514C/T多态性与冠心病相关性研究[J]。中国药理学通报,2006,22(9):1118-1121。
[52]姜红,杜明,西雁等。Fc Y受体Ⅱ A基因多态性与冠心病和扩张型心肌病的关系[J]。国际遗传学杂志,2006,29(4):246-249。
[53]梁茜,董吁钢,杨希立等。基因芯片技术分析老年冠心病患者的易感基因[J]。中华老年医学杂志,2006,25(6):520-523。
[54]Pereira AC,Miyakawa AA,Lopes NH,et al.Dynamic regulation of MTHFR mRNA expression and C677T genotype modulate mortality in coronary artery disease patients after revascularization[J].Thromb Res,2007;121(1):25-32.
[55]Satterthwaite G,Francis SE,Suvarna K,et al.Differential gene expression in coronary arteries from patients presenting with ischemic heart disease:further evidence for the inflammatory basis of atherosclerosis[J].Am Heart J,2005,150(3):488-499.
[56]O'Malley T,Ludlam C.A,Riemermsa R.A,et al.Early increase in levels of soluble inter-cellular adhesion molecule-1(sICAM-1)[J].European Heart Journal,2001(22),1226-1234.
[57]Atalar E,Aytemir K,Haznedaro(?)lu I,et al.Increased plasma levels of soluble selectins in patients with unstable angina[J].Int J Cardiol,2001,78(1):69-73.
[58]Inoue T,Komoda H,Nonaka M,et al.Interleukin-8 as an independent predictor of long-term clinical outcome in patients with coronary artery disease[J].Int J Cardiol,2008,124(3):319-325.
[59]Dominguez-Rodriguez A,Abreu-Gonzalez P,Garcia-Gonzalez M,et al.Prognostic value of interleukin-8 as a predictor of heart failure in patients with myocardial infarction and percutaneous intervention[J].Int J Cardiol,2006,111(1):158-160.
[60]Xiaoyong QI,Shuren LI,Junyong LI,et al.The prognostic value of IL-8 for cardiac events and restenosis in patients with coronary heart diseasesafter percutaneous coronary intervention[J].Jpn Heart J,2003,44(5):623-672.
[61]Xiaoyong QI,Junyong LI,Jian GU,et al.Plasma levels of IL-8 predict early complications in patients with coronary heart disease after percutaneous coronary intervention[J].Jpn Heart J,2003,44(4):451-461.
[62]Shimasaki Y,Saito Y,Yoshimura M,et al.The effects of long-term smoking on endothelial nitric oxide synthase mRNA expression in human platelets as detected with real-time quantitative RT-PCR[J].Clin Appl Thromb Hemost,2007,13(1):43-51.
[63]Shiraki R,Inoue N,Kobayashi S,et al.Toll-like receptor 4 expressions on peripheral blood monocytes were enhanced in coronary artery disease even in patients with low C-reactive protein[J].Life Sci,2006,80(1):59-66.
[64]Geng HL,Lu HQ,Zhang LZ,et al.Increased expression of Toll like receptor 4 on peripheral-blood mononuclear cells in patients with coronary arteriosclerosis disease[J].Clin Exp Immunol,2006,143(2):269-273.
[65]Willmar D,Patino,Omar Y,et al.Circulating transcriptome reveals markers of atherosclerosis[J].Proc Natl Acad Sci USA,2005,102(9):3423-3428.
[66]Anothai P,Roger R.T,Ian J,et al.LDL-Receptor mRNA expression in men is downregulated within an hour of an acute fat load and is influenced by genetic polymorphism[J].J Nutr,2007,137(9):2062-2067.
[67]Wang IJ,Chiang TH,Shih YF,et al.The association of single nucleotide polymorphisms in the MMP-9 genes with susceptibility to acute primary angle closure glaucoma in Taiwanese patients[J].Molecular Vision,2006,26(12):1223-1232.
[68]Pollin TI,Tanner K,Connell JR,et al.Linkage of plasma adiponectin levels to 3q27 explained by association with variation in the APM1 gene[J].Diabetes,2005,54(1):268-274.
[69]Damcott CM,Ott SH,Pollin TI,et al.Genetic variation in adiponectin receptor 1 and adiponectin receptor 2 is associated with type 2 diabetes in the old order Amish[J].Diabetes,2005,54(7):2245-2250.
[70]Akihiko M,Tomohiro N,Nobutaka D,et al.Polymorphism of the C-reactive protein (CRP) gene is related to serum CRP level and arterial pulse wave velocity in healthy elderly Japanese[J].Hypertens Res,2006,29(5):323-331.
[71]Russell Al,Cunninghame G DS,Shepherd C,et al.Polymorphism at the C-reactive protein locus influences gene expression and predisposes to-systemic lupus erythematosus[J].Hum Mol Genet,2004,13(1):137-147.
[72]Brull DJ,Serrano N,Zito F,et al.Human CRP gene polymorphism influences CRP levels:implications for the prediction and pathogenesis of coronary heart disease[J].Arterioscler Thromb Vasc Biol,2003,23(11):2063-2069.
[73]Zee RY,Ridker PM.Polymorphism in the human C-reactive protein (CRP) gene,plasma concentrations of CRP,and the risk of future arterial thrombosis[J].Atherosclerosis,2002,162(1):217-219.
[74]Szalai AJ,McCrory MA,Cooper GS,et al.Association between baseline levels of C-reactive protein (CRP) and a dinucleotide repeat polymorphism in the intron of the CRP gene[J].Genes Immun,2002,3(1):14-19.
[75]Dana C.C,Christopher L.S,Xiaoting Qin,et al.Genetic variation is associated with C-reactive protein levels in the third national health and nutrition examination survey[J].Circulation,2006,114 (23):2458-2465.
[76]Morita A,Nakayama T,Soma M.Association study between C-reactive protein genes and ischemic stroke in Japanese subjects[J].Am J Hypertens,2006,19 (6):593-600.
[77]Shen J,Arnett DK,Parnell LD,et al.Association of common C-reactive protein (CRP) gene polymorphisms with baseline plasma CRP levels and fenofibrate response:The GOLDN Study[J].Diabetes Care,2008,31(5):910-915.
[78]Kolz M,Koenig W,Müller M,et al.DNA variants,plasma levels and variability of C-reactive protein in myocardial infarction survivors:results from the AIRGENE study[J].Eur Heart J,2008,29(10):1250-1258.
[79]Mart(?)nez-Calatrava MJ,Gonz(?)lez-S(?)nchez JL,Mart(?)nez-Larrad MT,et al.Common haplotypes of the C-reactive protein gene and circulating leptin levels influence the interindividual variability in serum C-reactive protein levels[J].Thromb Haemost,2007,98(5):1088-1095.
[80]Wang XL,Mahaney MC,Sim AS,et al.Genetic contribution of the endothelial constitutive nitric oxide synthase gene to plasma nitric oxide levels[J].Arterioscler Thromb Vase Biol,1997,17(11):3147-3153.
[81]Tasic I,Milojkovic M,Sunder-Plassmann R,et al.The association of PC-1(ENPP1) K121Q polymorphism with metabolic syndrome in patients with coronary heart disease[J].Clin Chim Acta,2007,377(1-2):237-242.
[82]Hyun-Ju Seo,Soo-Geun Kim,Oh-Joong Kwon.The K121Q polymorphism in ENPP1(PC-1) is not associated with type 2 diabetes or obesity in Korean Male workers[J].J Korean Med Sci,2008,23(3):459-464.
[83]Jacek B,Grzegorz P,Krzysztof W,et al.New polymorphism of ENPP1(PC-1) is associated with increased risk of type 2 diabetes among obese individuals[J].Diabetes,2006,55(9):2626-2630.
[84]Kretowski A,McFann K,Hokanson JE,et al.Polymorphisms of the renin-angiotensin system genes predict progression of subclinical coronary atherosclerosis[J].Diabetes,2007,56(3):863-871.
[85]Guneri S,Baris N,Aytekin D,et al.The relationship between angiotensin converting enzyme gene polymorphism,coronary artery disease,and stent resteecNOSis[J].Int Heart J,2005,46(5):889-897.
[86]卢杨,屈会起。中国心肌梗死患者血管紧张素I转换酶基因插入/缺失多态性Meta分析[J]。循证医学,2001,1(1):18-21。
[87]李丽华,王曼丽,赵红等。冠心病患者血管紧张素转换酶基因多态性分析[J]。山东医药,2008,48(2):41-42。
[88]Stavroulakis GA,Makris TK,Krespi OG,et al.Predicting respose to chronic antihypertensivetreatment with fosinopril:the role of angiotensin converting enzyme gene polymorphism[J].Cardiovasc Drugs Ther,2000,14(4):427-432.
[89]L Tiret,B Rigat,S Visvikis,C Breda,et al.Evidence,from combined segregation and linkage analysis,that a variant of the angiotensin I-converting enzyme(ACE) gene controls plasma ACE levels[J].Am J Hum Genet,1992,51(1):197-205.
[90]Gordon V,Patrick P,Robert S,et al.Gene expression analysis of troglitazone reveals its impact on multiple pathways in cell culture:a case for in vitro platforms combined with gene expression analysis for early(idiosyncratic)toxicity screening[J].Inter J Toxicology,2006,25(2):85-94.
[91]Chen Q R,Gordon V,Kahuku O,et al.Diagnosis of the small round blue cell tumors using multiplex polymerase chain reaction[J].J Molecular Diagnostics,2007,9(1):80-88.
[02]Jean D.B,Tara M.R,Michele A.K,et al.Protection against simian/human immunodeficiency virus(SHIV) 89.6P in macaques after coimmunization with SHIV antigen and IL-15 plasmid[J].Proc Natl Acad Sci,2007,104(47):18648-18653.
[93]Vandesompele J,De P K,Pattyn F,et al.Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes[J].Genome Biol,2002,3(7):RESEARCH0034.
[94]Huggett J,Dheda K,Bustin S,et al.Real-time RT-PCR normalisation;strategies and considerations[J].Genes Immun,2005,6(4):279- 284.
[95]Tsuji N,Kamagata C,Furuya M,et al.Selection of an internal control gene for quantitation of mRNA in colonic tissues[J].Anticancer Res,2002,22(6C):4173-4178.
[96]Fuster G,Vicente R,Coma M,et al.One-step reverse transcription polymerase chain reaction for semiquantitative analysis of mRNA expression[J].Methods Find Exp Clin Pharmacol,2002,24(5):253-259.
[97]Ajay P S A,Sarvesh R,Sachinandan De,et al.Expression stability of two housekeeping genes(18S rRNA and G3PDH) during in vitro maturation of follicular oocytes in buffalo(Bubalus bubalis)[J].Animal Reproduction Science,2008,103(1-2):164-171.
[98]陈应泰,姜冠潮,王丹等。18srRNA作为内部参考在肺癌相关基因LSCC-3研究中的应用[J]。广西医科大学学报,2005,22(2):209-211。
[99]Pilbrow AP,Ellmers LJ,Black MA,et al.Genomic selection of reference genes for real-time PCR in human myocardium[J].BMC Med Genomics,2008,29;1 (1):64.
[100]Pierre V,Marc R,Tanveer A.K,et al.Differences in gene expression profiles of diabetic and nondiabetic patients undergoing cardiopulmonary bypass and cardioplegic arrest[J].Circulation,2004,110(2):280-286.
[101]Hendrik J.M.de J,Rudolf S.N.F,Eveline S.J.M.de B,et al.Evidence based selection of housekeeping genes[J].PLoS ONE,2007,2(9):e898.
[102]Teupser D,Mueller MA,Koglin J,et al.CD36 mRNA expression is increased in CD 14+ monocytes of patients with coronary heart disease[J].Clin Exp Pharmacol Physiol,2008,35(5-6):552-556.
[103]Scherrer-Crosbie M,UUrich R,Bloch KD,et al.Endothelial nitric oxide synthase limits left ventricular remodeling after myocardial infarction in mice[J].Circulation,2001,104(11):1286-1291.
[104]Jones SP,Greer JJ,van Haperen R,et al.Endothelial nitric oxide synthase overexpression attenuates congestive heart failure in mice[J].Proc Natl Acad Sci,2003,100(8):4891-4896.
[105]Akira K,Shin M,Takahiro M,et al.Increased expression of monocyte CD11a and intracellular adhesion molecule-1 in patients with initial atherosclerotic coronary stenosis[J].Circ J,2004;68(1):6-10.
[106]Bowen SL,Bloor BK,Leigh IM,et al.Adducin expression in cutaneous and oral lesions:alpha-and beta-adducin transcripts down-regulate with keratinocyte differentiation in stratified epithelia[J].J Pathol,2003,201 (1):119-126.
[107]Huijun S,Tomonari K,Tomonaga I,et al.C-reactive protein in atherosclerotic lesions[J].Am J Pathol.2005,167(4):1139-1148.
[108]Randall S,Sherri O L,Lisa H,et al.C-reactive protein downregulates endothelial NO synthase and attenuates reendothelialization in vivo in mice[J].Circulation Research,2007;100(10):1452-1459.
[109]Senthil K V,Sridevi D,Ivan Y,et al.Demonstration that C-reactive protein decreases eNOS expression and bioactivity in human aortic endothelial cells[J].Circulation,2002;106(12):1439-1441.
[110]Paolo C,James T.W,Edward T.H.Y,et al.Inflammatory cytokines stimulated C-reactive protein production by human coronary artery smooth muscle cells[J].Circulation,2003;108(16):1930-1932.
[1]Hristov M,Erl W,Weber PC.Endothelial progenitor cells:mobilization,differentiation,and homing.Arterioscler Thromb Vasc Biol,2003,23(7):1185-1189.
[2]Werner N,Kosiol S,Schiegl,et al.Circulating endothelial progenitor cells and cardiovascular outcomes.N Engl J Med,2005,353(10):999- 1007.
[3]Schmidt-Lucke C,Rossig L,Fichtlscherer S,et al.Reduced number of circulating endothelial progenitor cells predicts future cardiovascular events:proof of concept for the clinical importance of endogenous vascular repair.Circulation,2005,111(22):2981-2987.
[4]Blann AD,Woywodt A,Bertolini F,et al.Circulating endothelial cells.Biomarker of vascular disease.Thromb Haemost,2005,93(2):228-235.
[5]Massa M,Rosti V,Ferrario M,et al.Increased circulating hematopoietic and endothelial progenitor cells in the early phase of acute myocardial infarction.Blood,2005,105(1):199-206.
[6]George J,Goldstein E,Abashidze S,et al.Circulating endothelial progenitor cells in patients with unstable angina:association with systemic inflammation.Eur Heart J,2004,25(12):1003-1008.
[7]Adams V,Lenk K,Linke A,et al.Increase of circulating endothelial progenitor cells in patients with coronary artery disease after exercise-induced ischemia.Arterioscler Thromb Vasc Biol,2004,24(4):684-690.
[8]Fadini GP,Sartore S,Albiero M,et al.Number and function of endothelial progenitor cells as a marker of severity for diabetic vasculopathy.Arterioscler Thromb Vasc Biol,2006,26(9):2140-2146.
[9]Pirro M,Schillaci G,Paltriccia R,et al.Increased ratio of CD31+/CD42-microparticles to endothelial progenitors as a novel marker of atherosclerosis in hypercholesterolemia.Arterioscler Thromb Vasc Biol,2006,26(11):2530-2535.
[10]HenrichD,HahnP,WahlM,etal.Serum derived from multiple trauma patients promotes the differentiation of endothelial progenitor cells in vitro:possible role of transforming growth factor-betal and vascular endothelial growth factor165.Shock,2004,21(1):13-16.
[11]Kunz GA,Liang G,Cuculi F,et al.Circulating endothelial progenitor cells predict coronary artery disease severity.Am Heart J,2006,152 (1):190-195.
[12]Laufs U,Werner N,Link A,et al.Physical training increases endothelial progenitor cells,inhibits neointima formation,and enhances angiogenesis.Circulation,2004,109(2):220-226.
[13]Schomig K,Busch G,Steppich B,et al.Interleukin-8 is associated with circulating CD 133+ progenitor cells in acute myocardial infarction.Eur Heart J,2006,27(9):1032-1037.
[14]YoonYS,Johnson LA,ParkJS,et al.Therapeutic myocardial angiogenesis with vascular endothelial growth factors.Mol Cell Biochem,2004,264 (1-2):63-74.
[15]Kusuyama T,Omura T,Nishiya D,et al.The effects of HMG-CoA reductase inhibitor on vascular progenitor cells.J Pharmacol Sci,2006,101 (4):344-349.