Rapid detection of the cisAB allele consisting of a chimera of normal A and B alleles by PCR-RFLPs

Summary. DNA samples were analysed from Japanese individuals with the very rare ABO variant phenotype, cisAB (A₂B₃), which is characterized by the apparent inheritance of both A and B genes on one chromosome. The nature of the bases present at nucleotide positions (nps) 261, 526, 703, 796 and 803 is important for the specificity of the alleles at the ABO locus and the DNA from the cis AB donors was analysed by polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) to determine which nucleotides are present at these positions.
The results indicated that the cisAB allele had the AAAB-structure, which was a chimera of normal A and B alleles, when the expression 'AAAA' and 'BBBB' indicated the nucleotides of normal A (C, G, C and G) and B (G, A, A and C) genes at nps 526, 703, 796 and 803, respectively. The AAAB allele was found in all 27 individuals (17 families) with the cisAB including three phenotypes A₂B₃, A₁B₃ and A₂B and no other chimeric gene was found. The causative gene of cisAB was the AAAB allele, and the A and B alleles were not on one chromosome.
The cisAB allele appeared to be a product of the normal A allele due to a point mutation at nucleotide position 803, from G to C. The AAAB allele is thought to be normally transcribed and translated to produce an unusual transferase polypeptide, which has weak A- and weaker B-specific activity.
PCR-RFLP is a rapid and useful means of detecting the cisAB allele (the AAAB allele) without a family study, even when they have A₁B₃ and A₂B phenotypes, because trans-type A₁B₃ and A₂B samples have obviously different PCR-RFLP profiles.     

血管紧张素转换酶基因多态性与白族脑血管病患者的相关性研究

目的研究血管紧张素转换酶(ACE)基因插入/缺失多态性与云南大理白族脑血管病患者的相关性。方法应用聚合酶链式反应-限制性片段长度多态性(PCR-RFLP)技术,对73例白族脑血管病(ICVD)组患者(其中脑梗死40例,脑出血33例)和43例性别年龄相匹配的白族健康对照组进行ACE基因型检测和基因插入/缺失多态性分析;并进行ACE基因双向测序。结果(1)脑梗死组的DD基因型频率和D等位基因频率明显高于对照组(P<0.05);(2)脑出血组与对照组相比较,未发现DD基因型频率和D等位基因频率有明显差异(P>0.05);(3)脑血管组内伴高血压与不伴高血压,伴糖尿病与不伴糖尿病,伴血脂异常与血脂正常者相比较,未发现DD基因型频率和D等位基因频率有显著差异(P>0.05);(4)D等位基因和I等位基因其核苷酸序列和长度与国内外文献报道无明显差别。结论(1)ACE基因插入/缺失多态性与脑梗死的发生有关联性,DD基因型和D等位基因是脑梗死患者的高危因素;(2)ACE基因插入/缺失多态性与脑出血发生未发现有关联性;(3)脑血管病的其他相关危险因素如高血压,糖尿病,血脂与ACE基因多态性可能无关联;(4)ACE基因16内含子...     

宁夏汉族人126名甘露糖结合凝集素基因多态性分析

目的检测宁夏汉族人群中甘露糖结合凝集素(MBL)基因多态性,获得数据为进一步研究MBL基因突变与疾病间的关联性提供依据。方法采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)方法,检测126名宁夏汉族健康个体的MBL基因3个多态性位点,并与广东汉族人群MBL基因多态性的分布进行比较。结果在宁夏汉族人群中,只检测出两种等位基因:野生型A和变异型B,未检出变异型C、D等位基因,同时发现MBL基因的基因频率A为0.853 2,B为0.146 8,基因型频率分别是AA为0.714 3,AB为0.277 8,BB为0.007 9。结论宁夏汉族MBL基因多态性符合Hardy-Weinberg平衡定律,具有群体代表性,与文献中的广东汉族人群相比较有一定的差异。     

IRS-4和GCKR基因多态性与2型糖尿病的关联性研究

目的 研究IRS-4和GCKR基因多态性与黑龙江汉族2型糖尿病的关联性.方法 采用PCR-RFLP技术检测黑龙江汉族2型糖尿病组、正常对照组人群的IRS-4基因密码子879C/G和GCKR基因P446L多态性,并分析其基因型频率及等位基因频率.结果 2型糖尿病组IRS-4基因879C/G的GG、GC、CC基因型频率分别为0.538、0.404、0.058,G及C等位基因频率分别为0.740、0.260;正常对照组IRS-4基因879C/G的GG、GC、CC基因型频率分别为0.556、0.352、0.092,G及C等位基因频率分别为0.732、0.268;糖尿病组和正常对照组之间IRS-4基因879C/G基因型频率及等位基因频率差异无统计学意义(P＞0.05).2型糖尿病组GCKR基因P446L的PP、PL、LL基因型频率分别为0.212、0.596、0.192,P和L等位基因频率分别为0.510、0.490;正常对照组GCKR基因P446L的PP、PL、LL基因型频率分别为0.148、0.463、0.389,P和L等位基因频率分别为0.380、0.620;糖尿病组和正常对照组之间GCKR基因P446L基因型频率及等位基因频率差异有统计学意义(P＜0.01).结论 IRS-4基因879C/G多态性与黑龙江省汉族2型糖尿病的发生无相关性,GCKR基因P446L多态性与黑龙江省汉族2型糖尿病的发生有相关性.     

中国2型糖尿病人中CYP2C8、CYP2C9基因多态性

目的：查明CYP2C9、CYV2C8等抗糖尿病药物主要代谢酶的基因多态性在中国2型糖尿病（T2DM）A-群中的分布频率和分布特征。方法：运用多聚酶链反应．限制性长度多态性（PCR．RFLP）方法和变性高效液相色谱法（DHPLC）对222名中国T2DM患者进行了有功能意义的CYP2C8＊3、CYP2C8P404A和CYP2C9＊3,以及可能存在功能意义的CYP2C8IVs2（-5insertt）突变体等等位基因型检测,并计算了各等位基因的频率。结果：222名中国T2DM患者的CYP2C8基因中未检出CYP2C8＊3、P404A型,CYP2C8IVS2（-5insertt）和CYP2C9＊3等位基因的频率分别为43．0％、2．48％,二者突变等位基因在男、女性别分布中不存在差异（P〉0．05）,同时为CYP2C8IVS2（-5insertt）和CYP2C9＊1＊3基因的频率为6．3％,该联合突变基因型的分布也不存在性别差异（P〉0．05）。结论：中国T2DM患者中CYP2C9＊3的等位基因频率为2．48％;未检出CYP2C8。3和P404A型,CYP2C81VS2（-5insertt）突变体发生频率为43．0％,其是否影响CYV2C8的代谢活力有待于进-步研究。     

Ile<Superscript>105</Superscript>Val <Emphasis Type="Italic">GSTP1</Emphasis> polymorphism and susceptibility to colorectal carcinoma in Bulgarian population

Background and aims Etiologically, the sporadic colorectal cancer (CRC) is a complex and multifactorial disease that is linked to both exogenic and endogenic factors. Accumulating evidence indicates that susceptibility to cancers, including CRC, is mediated by genetically determined differences in the effectiveness of detoxification of potential carcinogens. A member of the glutathione-S-transferase (GST) family, GSTP1, is an important candidate for involvement in susceptibility to carcinogen-associated CRC. An A→G transition in exon 5 of the GSTP1 gene resulting in Ile¹⁰⁵Val amino acid substitution has been identified. This change leads to alteration in catalytic efficiency of variant enzyme. The aim of the current study was to evaluate the influence of Ile¹⁰⁵Val GSTP1 polymorphism on susceptibility to CRC. Materials and methods The GSTP1 genotyping was conducted in a case-control study of 80 ethnic Bulgarian CRC patients and 126 unaffected controls using polymerase chain reaction restriction fragment length polymorphism method. Results A statistically significant case-control difference in genotype frequencies was observed: 0.69 vs 0.54 for Ile/Ile, 0.22 vs 0.39 for Ile/Val, and 0.09 vs 0.07 for Val/Val (p = 0.049). The odds ratio (OR) for Val/Val was close to 1 (0.96, 95%CI: 0.35–2.66, p = 0.942), whereas the OR for Ile/Val was significantly lower, 0.45 (95%CI: 0.24–0.86, p = 0.016), compared to the referent Ile/Ile genotype. Although a prevalence of the GSTP1 variant allele-containing genotypes (Ile/Val or Val/Val) was found in controls than in patients (OR = 0.53, 95%CI: 0.30–0.96, p = 0.035), the allele frequencies did not show significant difference between cases and controls (p = 0.127). Conclusions Based on the obtained protective effect of Ile/Val GSTP1 genotype, we could suggest that Ile¹⁰⁵Val GSTP1 polymorphism may play some role in susceptibility to CRC.     

云南滇南小耳猪与广西巴马小型猪mtDNA D-Loop的初步研究

目的研究封闭群滇南小耳猪与广西巴马小型猪线粒体DNA（mtDNA）的多态性,探索小型猪遗传监测的方法。方法用18种限制性内切酶对23头滇南小耳猪和21头广西巴马小型猪mtDNA D-loop的PCR产物进行RFLP分析。结果滇南小耳猪和广西巴马小型猪的mtDNA D—loop在品种内和品种间均表现出相同的酶切图谱,未见多态性。结论滇南小耳猪与广西巴马小型猪mtDNA D—loop的遗传多样性贫乏,mtDNA PCR—RFLP技术在其遗传监测中不具应用价值。     

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.     

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。     

新疆哈萨克及维吾尔族SLE和RA患者MBL基因多态性调查

目的 对新疆哈萨克及维吾尔族人群中系统性红斑狼疮(SLE)及类风湿关节炎(RA)患者甘露糖结合凝集素基因54位密码子点突变进行测定并分析.方法 PCR-RFLP.结果 42例哈萨克族SLE患者基因频率:0.536(GGC),0.464(GAC);24例维吾尔族SLE患者基因频率:0.667(GGC),0.333(GAC);22例哈萨克族RA患者基因频率:0.614(GGC),0.386(GAC);15例维吾尔族RA患者基因频率:0.733(GGC),0.267(GAC).结论 MBL不同的基因型和这些疾病发生可能存在相关.