apo（a）PNR基困多态性在汉族人群中的表达与冠心病的关系

目的：通过检测冠心病患者和正常人群apo(a)PNR基因型及血脂水平的变化，研究载脂蛋白(a)[apo(a)]五核苷酸重复序列(PNR)基因多态性与冠心病(CHD)及其血脂水平的关系。方法：采用聚合酶链反应结合高压变性聚丙烯酰胺凝胶电泳，分析了153例健康志愿者及88例冠心病患者(武汉大学中南医院，武汉大学亚太医院1998—1999住院患者)的apo(a)PNR基因型，同时检测基因型与血清脂质。脂蛋白和载脂蛋白水平之间的关系。结果：CHD组apo(a)PNR(TTTTA)5/8基因型频率(0．181)和(TTTTA)5等位基因频率(0．107)分别显著高于对照组(0．039，χ^2=13．779，P&;lt;0．01；0．026，r=6．39，P&;lt;0．05)；(TTTTA)5/8基因型者血清LP(a)水平高于(TTTTA)8/8，(TTTTA)8/9基因型者。结论：apo(a)PNR基因多态性与Lp(a)水平密切相关，apo(a)PNR(TTTTA)5等位基因可能是中国汉族人群冠心病的危险因子之一。此结论可为冠心病的康复、预防提供理论依据。     

脂蛋白(a)、载脂蛋白(B)基因多态性与 心肌梗死的相关性研究

目的了解脂蛋白(a)、载脂蛋B基因多态性与心肌梗死的关系。方法用免疫透射比浊法测定血清脂蛋白(a)。采用聚合酶链反应/限制性酶切法(PCR)/RFLP)分析ApoB基因。结果正常健康人组脂蛋白(a)(170±148)mg/L,ApoBX     

原发性肾病综合征患儿载脂蛋白(a)(TTTTA)n重复序列基因多态性与脂蛋白a水平的关系

目的　探讨Apo(a)五核苷酸重复序列基因多态性 (PNTR)在原发性肾病综合征 (PNS)中的分布情况及特点 ,并分析其与血清中Lp(a)水平的关系。方法　采用聚合酶链反应 (PCR)结合非变性聚丙烯酰胺凝胶电泳法检测了 6 0例肾病综合征患儿及 30例健康对照组Apo(a) (TTTTA)n基因多态性 ;并测定血清中脂蛋白a水平。结果　两组人群Apo(a)PNTR共检出 6种等位基因和 12种基因型。PNS组 (TTTTA) 5-7频率明显高于对照组 (P <0 0 5 )。原发性肾病综合征患者Lp(a)水平明显高于对照组 ,增高水平与TTTTA重复次数呈负相关。结论　原发性肾病综合征患者血清中Lp(a)水平与载脂蛋白 (a) (TTTTA)n基因多态性有关。     

apo(a)表型与维持性透析患者高Lp(a)水平关系研究

目的 研究载脂蛋白（a）[apo（a）]表型与维持性透析患者高脂蛋白（a）[Lp（a）]水平关系,探索血液透析患者Lp（a）代谢紊乱机制。方法 维持性透析患者66例,持续性透析治疗两年以上。终末期肾病患者51例,未经过血液透析、腹膜透析及肾移植治疗。随机选择62例体检健康者作为对照组。应用SDS-琼脂糖电泳连接蛋白免疫印迹方法检测各组患者apo（a）表型,免疫透射比浊法检测Lp（a）。结果 HMW apo（a）表型所占比例在MHD组、ESRD组与对照组三组中无显著性差异（P〉0.05）。在HMW apo（a）表型中,Lp（a）中位数浓度在MHD组、ESRD组与对照组分别为100.9mg/L8、3.5 mg/L5、6.5 mg/L,MHD组与ESRD组及对照组之间都存在显著性差异（P〈0.05）。对于LMW apo（a）表型,MHD组和ESRD组Lp（a）中位数浓度分别为220.5 mg/L2、15.8 mg/L,无显著性差异（P〉0.05）,但两组分别与对照组（163.0mg/L）相比,则均有显著性差异（P〈0.05）。结论 MHD治疗可促使Lp（a）浓度增加,并且以HMW apo（a）表型的Lp（a）浓度增高更为显著。     

冠心病ApoE基因多态性及其与血清ApoE的关系

目的探讨冠心病(Coronary Heart Disease, CHD)ApoE基因多态性及其与血清ApoE的关系.方法用银染PCR-RFLP法来检测ApoE基因多态性,其中健康对照者121例,CHD患者120例,测定所有样本的血清ApoE水平.结果 1. CHD组中E3/4基因型(含E4/4)含量(21)较对照组(10)高(P<0.05);CHD患者携带ε 4等位基因(31)较对照组(14)高(P<0.01).2. CHD组血清ApoE[(45.50±18.24) mg/L]明显高于对照组[(35.42±8.89) mg/L](P<0.001).3. CHD组E2/3(含E2/2)的ApoE水平[(53.34±22.0) mg/L]明显高于E3/3[(44.14±14.9) mg/L](P<0.001).CHD组E3/4(含E4/4的)ApoE水平[(34.42±8.37) mg/L]明显低于E3/3[(44.14±14.9) mg/L](P<0.001).CHD组E4/4的ApoE水平[(28.06±5.81) mg/L]明显低于E3/4[(38.12±8.56) mg/L](P<0.05).结论ε 4等位基因与CHD密切相关;ApoE基因多态性可能是通过影响ApoE水平而实现其CHD致病作用.     

冠心病ApoE基因多态性及其与血清ApoE的关系

目的　探讨冠心病 (CoronaryHeartDisease ,CHD)ApoE基因多态性及其与血清ApoE的关系。 方法　用银染PCR RFLP法来检测ApoE基因多态性 ,其中健康对照者 12 1例 ,CHD患者 12 0例 ,测定所有样本的血清ApoE水平。 结果　 1.CHD组中E3/ 4基因型 (含E4 / 4 )含量 (2 1)较对照组 (10 )高 (P <0 .0 5 ) ;CHD患者携带ε 4等位基因 (31)较对照组 (14 )高 (P <0 .0 1)。 2 .CHD组血清ApoE[(45 .5 0± 18.2 4 )mg/L]明显高于对照组[(35 .4 2± 8.89)mg/L](P <0 .0 0 1)。 3.CHD组E2 / 3(含E2 / 2 )的ApoE水平 [(5 3.34± 2 2 .0 )mg/L]明显高于E3/ 3[(44 .14± 14 .9)mg/L](P <0 .0 0 1)。CHD组E3/ 4 (含E4 / 4的 )ApoE水平 [(34.4 2± 8.37)mg/L]明显低于E3/ 3[(44 .14± 14 .9)mg/L](P <0 .0 0 1)。CHD组E4 / 4的ApoE水平 [(2 8.0 6± 5 .81)mg/L]明显低于E3/ 4[(38.12± 8.5 6 )mg/L](P <0 .0 5 )。 结论　ε 4等位基因与CHD密切相关 ;ApoE基因多态性可能是通过影响ApoE水平而实现其CHD致病作用     

脂蛋白（a）、载脂蛋白（B）基因多态性与心肌梗死的相关性研究

目的了解脂蛋白 (a)、载脂蛋 B基因多态性与心肌梗死的关系。方法用免疫透射比浊法测定血清脂蛋白 (a)。采用聚合酶链反应 /限制性酶切法 (PCR) /RFL P)分析 Apo B基因。结果正常健康人组脂蛋白 (a) (170±148) m g/L ,Apo BX 等位基因频率为 0 .0 82 ;高血压组脂蛋白 (a) (4 82± 15 1) mg/L ,Apo BX 等位基因为 0 .0 30 ;心肌梗死组脂蛋白 (a) (781± 2 78) mg/L ,Apo BX 等位基因频率为 0 .30 2。结论脂蛋白 (a)的水平 (≥ 30 0 0 m g/L ) ,Apo BX 等位基因频率与心肌梗死高度相关 ,高血压组的脂蛋白 (a)、Apo BX 等位基因频率与正常健康人相比较无明显差异。     

脂蛋白(a)的多态性

本文主要综述脂蛋白(a)的一般生理特点、多态性及其临床意义。     

昆明地区汉族人群载脂蛋白B基因多态性频率分布与高脂血症关系研究

目的探讨昆明地区汉族人群载脂蛋白B(ApoB)基因中,MspI、XbaI、EcoRI基因型及等位基因频率分布及其与原发性高脂血症的相关性。方法采用病例-对照相关性研究策略,选择昆明地区汉族人群作为研究对象,对91例高脂血症患者及76例健康对照者进行基因多态性检测。结果 1)76例健康者中,MspI位点的+/+、+/-基因型频率分别是0.618、0.382,未检出-/-基因型;+和-的等位基因频率分别是0.809、0.191;XbaI位点的+/+、+/-、-/-基因型频率分别为0.013、0.145、0.842;+和-等位基因频率分别是0.086、0.914;EcoRI位点的+/+、+/-基因型频率分别是0.961、0.039,未检出-/-基因型;+和-等位基因频率分别是0.980、0.020。2)91例高脂血症患者中,XbaI位点的+/+、+/-及-/-基因型和等位基因频率与对照组比较,差异无统计学意义(P0.05);MspI位点+/+基因型频率高于对照组,差异有统计学意义(P0.05),Odd Ratio=27.40(95%CI:6.28～120.12);EcoRI位点的+/+基因型频率低于对照组,差异有统计学意义(P0.05),Odd Ratio=0.247(95%CI:0.068～0.901)。结论 1)昆明地区汉族健康人群ApoB基因MspI、XbaI、EcoRI基因多态性有地区特征;2)该地区汉族高脂血症患者中的MspI基因位点的+/+基因型与高脂血症易感性相关,而EcoRI基因位点+/+基因型与高脂血症发病保护性可能相关。     

载脂蛋白E基因多态性与汉族人群心肌梗死的相关性

心肌梗死(MI)是一种复杂的综合征,同时受到多种遗传基因和环境因素的影响,基本病因是冠状动脉粥样硬化。对于载脂蛋白E(ApoE)基因多态性和MI危险因素之间的关系已经研究了近40年,仍然未能达成一致的观点。近几年,MI与基因-基因、基因-环境的相互作用越来越被学者所认可,因此,ApoE基因多态性也成为全球学者研究的热点。     

The association of serum lipoprotein(a) levels, apolipoprotein(a) size and (TTTTA)(n) polymorphism with coronary heart disease.

BACKGROUND: The association between lipoprotein(a) levels, apolipoprotein(a) size and the (TTTTA)(n) polymorphism which is located in the 5' non-coding region of the apo(a) gene was studied in 263 patients with severe coronary heart disease and 97 healthy subjects. METHODS: Lp(a) levels were measured by ELISA, apo(a) isoform size was determined by SDS-agarose gel electrophoresis, and analysis of the (TTTTA)(n) was carried out by PCR. For statistical calculation, both groups were divided into low (at least one apo(a) isoform with < or = 22 Kringle IV) and high (both isoforms with >22 KIV) apo(a) isoform sizes, and into low number (<10 in both alleles) and high number of (> or =10 at least one allele) TTTTA repeats. RESULTS: Lp(a) levels were higher (P=0.007), apo(a) isoforms size < or =22 KIV and TTTTA repeats > or = 10 were more frequent (P=0.007 and 0.01) in cases than in controls. Lp(a) levels were found to be increased with low apo(a) weight in both groups (both P<0.0001). In multivariate logistic regression analysis, only the Lp(a) levels (P=0.005) and (TTTTA)(n) polymorphism (P=0.002) were found to be significantly associated with CHD. CONCLUSION: Nevertheless, these results indicate that in CHD patients the (TTTTA)(n) polymorphism has an effect on Lp(a) levels which is independent of the apo(a) size.     

载脂蛋白（a）五核苷酸患联重复序列基因型的检测

目的 建立一种简便、快速、准确、实用的人载脂蛋白（ａ）五核苷酸串联重复序列（ｐｅｎｔａｎｕｃｌｅｏｔｉｄｅ ｒｅｐｅａｔｓ,ＰＮＲ）基因型的检测方法。方法 应用聚合酶链反应（ＰＣＲ）特异性扩增Ａｐｏ（ａ）５′端调控区ＰＮＲ,扩增产物用６％的变性聚丙烯酰胺凝胶电沪结合硝酸银染色,观察Ａｐｏ（ａ）ＰＮＲ的多态性。结果 应用该法检测了１５３例健康人Ａｐｏ（ａ）５′端调控区ＰＮＲ基因型,共检出７种等位基因     

血清胆红素和血脂水平与冠心病的相关性分析

目的探讨血清胆红素水平和血脂变化与冠心病(CHD)的关系,为CHD的预防、治疗提供临床依据。方法以87例CHD患者作为观察组,87例健康体检者为健康对照组,分别检测其血清总胆红素(TBIL)、直接胆红素(DBIL)、总胆固醇(TC)、三酰甘油(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)水平,并进行比较分析。结果 CHD观察组对比健康对照组血清TBIL、DBIL、HDL-C浓度明显降低,TC、TG、LDL-C浓度CHD观察组明显增高,差异具有统计学意义(P<0.05)。结论 TBIL、DBIL、HDL-C浓度降低和TC、TG、LDL-C浓度增高是影响CHD发生和发展的危险因素。     

Apolipoprotein B gene 3'VNTR polymorphism: association with plasma lipids and coronary heart disease in Han Chinese.

BACKGROUND: Studies that considered polymorphisms within the apolipoprotein B (APOB) gene as risk factors for coronary heart disease (CHD) have reported conflicting results. METHODS: The phenotypic effects of the 3'VNTR polymorphism of the APOB gene on the susceptibility to CHD were investigated in 120 unrelated healthy individuals and 137 CHD patients. The internal structure of APOB gene 3'VNTR alleles was also analyzed by the methods of SspI restriction mapping and DNA sequencing of the allele fragments. RESULTS: In total, 14 segregating alleles and 32 genotypes of APOB gene 3'VNTR were characterized in the pooled total of 257 subjects. The frequency of 3'VNTR-B alleles [hypervariable element (HVE) > or =38)] in the CHD cases was higher than that of the controls (10.95% vs. 5.00%, p<0.05). 3'VNTR-B allele was dependently related to total cholesterol levels (p<0.05). Compared with SS homozygotes, 3'VNTR-B allele carriers were associated with an increased risk of CHD (OR=2.137, 95% CI=1.055-4.328, p=0.0349). No significant differences in the internal structure and sequences of APOB gene 3'VNTR alleles were found between cases and controls. CONCLUSIONS: APOB gene 3'VNTR polymorphism exerts an impact on lipid metabolism and may contribute to the susceptibility to the development of CHD in Han Chinese.     

Fluorescence-based, nonradioactive method for efficient detection of the pentanucleotide repeat (TTTTA)(n) polymorphism in the apolipoprotein(a) gene

BACKGROUND: The apolipoprotein(a) [apo(a)] gene is a major predictor of plasma lipoprotein(a) concentrations, an independent risk factor for cardiovascular disease. The apo(a) gene contains a pentanucleotide repeat (PNR) polymorphism, 1.4 kb upstream from the apo(a) gene reading frame. This polymorphism has been suggested to be important in control of apo(a) gene expression. METHODS: We developed a fluorescence-based, nonradioactive procedure to detect the PNR polymorphism. After amplification of the polymorphism by PCR, the respective PCR products were separated by denaturing polyacrylamide gel electrophoresis and detected using a 3'-end fluorescently labeled oligonucleotide as a probe. We used the method to characterize the PNR polymorphism pattern in 313 individuals, 195 Caucasians and 118 African Americans. The new method efficiently separated DNAs corresponding to the different PNR repeats. RESULTS: Among both ethnic groups, alleles containing eight PNRs were most common. Smaller PNRs were more common among African Americans, and larger PNRs were more common among Caucasians. CONCLUSIONS: We developed a nonradioactive technique that separates the PNR polymorphism in the apo(a) gene and can be used in other studies involving closely sized polymorphisms.     

A pentanucleotide repeat polymorphism in the 5' control region of the apolipoprotein(a) gene is associated with lipoprotein(a) plasma concentrations in Caucasians.

The enormous interindividual variation in the plasma concentrations of the atherogenic lipoprotein(a) [Lp(a)] is almost entirely controlled by the apo(a) locus on chromosome 6q26-q27. A variable number of transcribed kringle4 repeats (K4-VNTR) in the gene explains a large fraction of this variation, whereas the rest is presently unexplained. We here have analyzed the effect of the K4-VNTR and of a pentanucleotide repeat polymorphism (TTTTA)n (n = 6-11) in the 5' control region of the apo(a) gene on plasma Lp(a) levels in unrelated healthy Tyroleans (n = 130), Danes (n = 154), and Black South Africans (n = 112). The K4-VNTR had a significant effect on plasma Lp(a) levels in Caucasians and explained 41 and 45% of the variation in Lp(a) plasma concentration in Tyroleans and Danes, respectively. Both, the pentanucleotide repeat (PNR) allele frequencies and their effects on Lp(a) concentrations were heterogeneous among populations. A significant negative correlation between the number of pentanucleotide repeats and the plasma Lp(a) concentration was observed in Tyroleans and Danes. The effect of the 5' PNRP on plasma Lp(a) concentrations was independent from the K4-VNTR and explained from 10 to 14% of the variation in Lp(a) concentrations in Caucasians. No significant effect of the PNRP was present in Black Africans. This suggests allelic association between PNR alleles and sequences affecting Lp(a) levels in Caucasians. Thus, in Caucasians but not in Blacks, concentrations of the atherogenic Lp(a) particle are strongly associated with two repeat polymorphisms in the apo(a) gene.     

人载脂蛋白基因A5 c.553 G＞T多态性与冠心病及血脂水平的相关性研究

目的分析中国江苏地区汉族人群载脂蛋白（apo）A5c．553G〉T基因多态性与冠心病及血脂水平的关系。方法采用聚合酶链反应-限制性片段长度多态性（PCR—RFLP）技术分别检测201例冠心病患者和222名健康对照者apoA5c．553G〉T多态性基因型和等位基因的分布，并采用生化方法检测研究对象的血脂水平。结果健康对照组和冠心病组apoA5基因c．553G〉T位点GG、GT、TT基因型分别为91．89％、8．11％、0％和82．58％、16．92％、0．50％（Χ^2=9．257，P=0．010）；T等位基因频率分别为4．05％和8．96％（Χ^2=8．481，P=0．004）。冠心病组T等位基因携带者血清三酰甘油和总胆固醇水平分别为（2．17±1．05）mmol／L和（5．16±0．95）mmol／L，均明显高于GG基因型人群（t=3．194和3．022；P=0．002和0．003）。此外，T等位基因携带者患冠心病的风险较G等位基因携带者高出2．39倍（OR=2．39，95％CI 1．31～4．37，P=0．005），经Logistic多元回归校正年龄、性别、体重指数、高血压、吸烟史、糖尿病史其他冠心病危险因素后，差异仍具有统计学意义（OR=2．25，95％CI1．18～4．30，P=0．014）。结论江苏地区汉族人群apoA5C．553G〉T基因多态性与冠心病存在一定的关联性，并影响血清三酰甘油和总胆固醇水平。     

载脂蛋白A5基因多态性与冠心病的相关性研究

目的研究太原汉族人群中载脂蛋白A5(ApoA5)-1131T/C基因多态性与冠心病(CHD)的关系。方法采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)检测了249例CHD患者和176例健康人群的ApoA5-1131T/C的多态性基因型和等位基因的分布。结果两组间ApoA5-1131T/C等位基因和基因型频率存在明显差异,等位基因C的频率在CHD组显著低于对照组(37.1%VS42.0%,P<0.05)。结论 ApoA5的基因多态1131T/C与CHD的发病率有一定的相关性(P<0.05)。     

汉族人凝血酶活化的纤溶抑制物编码区的基因多态性与冠状动脉粥样硬化性心脏病相关性研究

目的研究汉族人血浆中凝血酶活化的纤溶抑制物水平及编码区的基因多态性分布特征与冠状动脉粥样硬化性心脏病之间的相关性。方法应用ELISA及发色底物法分别检测了126例冠状动脉粥样硬化性心脏病患者和45名健康对照组血浆中的TAFI抗原及活性水平，同时应用PCR结合限制性片段长度多态性技术研究了浙江地区156名汉族健康人与126例冠状动脉粥样硬化性心脏病患者TAFI基因编码区Thr325Ile、Thrl47Ala多态性分布特点，并分析这两个多态性位点与冠状动脉粥样硬化性心脏病之间的关系。结果急性心肌梗死组患者血浆中TAFI抗原与活性水平分别为（106．82±24．51）％和（6．32±1．58）μg／ml，心绞痛组患者血浆中TAFI抗原与活性水平分别为（100．73±30．39）％和（6．92±1．78）μg／ml，经方差分析，二者之间及其与对照组之间的差异无统计学意义（P〉0．05）；在浙江汉族人群中TAFI Thr325Ile的基因多态性分布中Thr325Thr（1040C／C）占0．6670，Thr325Ile（1040C／T）占0．2620，Ile325Ile（1040T／T）占0．0710；1040位C与T等位基因频率的分别为0．7980与0．2020，TAFI Thrl47Ala的基因型分布中Alal47Ala（505G／G）为0．6030，Ala147 Thr（505G／A）为0．2780，Thrl47Thr（505A／A）为0．1190，505位G与A等位基因频率分别为0．7420与0．2580，冠状动脉粥样硬化性心脏病组与对照组中基因型的分布频率和等位基因频率之间的差异也无统计学意义（P〉0．05）；另外，在Thr325Ile基因多态性中，纯合子Thr325Thr（1040C／C）者血浆TAFI抗原水平为（114．89±22．53）％，较其他两型（1040C／T、1040T／T）高，差异有统计学意义（P〈0．05），而后两型之间差异无统计学意义（P〉0．05）；纯合子Ile325Ile（1040T／T）的TAFI活性为（3．08±3．63）μg／ml，较其他两型（1040C／T、1040T／T）低，差异有统计学意义（P〈0．05），而后两者之间的差异无统计学意义（P〉0．05）；而Thrl47Thr（505A／A）、Thrl47Ala（505C／T）、Alal47Ala（505G／G）3种基因型血浆中TAFI水平（抗原与活性）之间的差异均无统计学意义（P〉0．05）。结论TAFI编码区Thr325Ile基因多态性对血浆中TAFI抗原与活性水平有明显的影响，但TAFI Alal47Thr与Thr325Ile两种基因多态性与冠状动脉粥样硬化性心脏病的发生无明显的相关性。