昆明地区汉族人vWF基因多态性与急性脑梗死的相关性

目的 探讨血浆血管性假性血友病因子(von Willebrand factor vWF)表达水平及其基因19内含子MspI(Major surface protease I)多态性与急性脑梗死的相关性.阐明vWF基因19内含子MspI多态性在昆明地区汉族人群中的分布.方法 收集119例急性脑梗死(24 h内)和117名昆明地区汉族健康人为对照,免疫比浊法检测血浆vWF 表达水平;聚合酶链反应-限制性片段长度多态性结合MspI酶切技术进行vWF基因19内含子MspI多态性分析.结果 急性脑梗死血浆vWF表达水平为(189.37% ± 89.55%)明显高于对照组(P ＜ 0.05).昆明地区汉族人群中vWF基因19内含子MspⅠM+/M+、M+/M-和 M-/M-基因型频率分布分别为0.047、0.377和0.576;M+/M-等位基因型频率分布分别为0.235、0.765.脑梗死组MspⅠ多态性M-/M-基因型频率高于对照组(P ＜ 0.05).脑梗死组M+/M+、M+/M-和M-/M-型的循环vWF 水平分别为(170.00% ± 78.67%)、(148.62% ± 62.58%)和(205.38% ± 92.68%),各基因型之间差异有统计学意义(P ﹤0.05).结论 血浆vWF表达水平及vWF 基因19 内含子MspⅠ多态性M-/M-基因型与急性脑梗死发生有显著相关性;昆明地区汉族人群中vWF基因19内含子MspⅠ等位基因M+和M-的分布与其他人群不同.     

高甘油三酯血症患者载脂蛋白B基因的MspI多态性研究

利用Apo B cDNA探针（LB 1.5）,检测13名高甘油三酯血症（HTG）患者和21名正常对照者的Apo B基因MspI限制性片段长度多态性（RFLP）等位基因频率。研究结果显示,34份标本共出现2.35kb（M_1）和2.60kb（M_2）二种不同长度的杂交片段。M_2等位基因频率在HTG患者（0.192）和对照者（0.024）之间存在明显差异（P=0.02）。     

Identification of Candida species using PCR-RFLP in cancer patients in Iran

Opportunistic infections caused by Non–Candida albicans. have been increasing. Traditional methods that are used to identify clinical isolates of Candida species are time-consuming and not appropriate for rapid, accurate and reliable identification. Purpose: To identify Candida spp isolated from cancer patients using PCR-restriction enzyme. Materials and ethods: Using universal primers, ITS1 and ITS4, in this study, we could amplify ITS1-5.8S-ITS2 rDNA regions at both 80 clinical isolates and 3 standard strains. The PCR products were digested with two restriction enzymes MspI and BlnI separately. Result: We successfully identified all isolated species using two restriction enzymes (MspI, BlnI). Candida albicans was the most common species (77.5%), followed by C. glabrata (15%), C. tropicalis (5%), C. krusei (2.5%). Although the primers and enzyme had the ability to identify C. parapsilosis, C. guilliermondii, C. dubliniensis, present isolates did not include these among identified ones. Conclusion: RFLP-PCR using ITSI and ITS4 primers and restriction enzyme is a rapid, easy, reliable and also applicable method in clinical laboratory for identification of medically important Candida spp.     

The Relationship between Aryl Hydrocarbon Hydroxylase and Polymorphisms of the CYP1A1 Gene

We examined the relationship between aryl hydrocarbon hydroxylase (AHH) and the frequency of a Msp I mutation in the 3'-flanking region of cytochrome P450 (CYP) 1A1 (Mspl polymorphism) and another mutation in exon 7 (Ile-Val polymorphism) in 84 healthy male subjects in Fukuoka, Japan. AHH inducibility (3-methylcholanthrene (MC)-induced AHH activity/non-induced AHH activity) was correlated with the MspI polymorphism (P<0.0001) and age class (P=0.015), whereas no correlation was found for the Ile-Val polymorphism (P=0.509). Age-adjusted AHH inducibility (mean±SE) of the predominant homozygote (genotype A), the heterozygote (genotype B) and a homozygote rare allele (genotype C) genotypes was 4.89±0.36, 4.82±0.29 and 13.61±1.44, respectively. The genotype C showed much higher AHH inducibility than genotypes A and B (P<0.001), while no significant difference was observed between genotypes A and B. Non-induced AHH activity was also correlated with these polymorphisms. The AHH activity of a homozygous mutant Val/Val genotype (0.076±0.010 pmol/min/10⁶ cells) was significantly higher (P<0.05) than that of the wild-type homozygous Ile/Ile (0.044±0.004 pmol/min/10⁶ cells) and heterozygous Ile/Val (0.047±0.007 pmol/min/10⁶ cells) genotypes. Our study suggests that the genotypes C and Val/Val, which are more frequent in smoking-related lung cancer, are closely related with high AHH inducibility and high non-induced AHH activity, respectively. Thus, the positive relationship between AHH inducibility and lung cancer is supported by our study. If our results are confirmed and the assessment of genotype becomes feasible on a population basis, identification of smokers who have genetically high susceptibility to lung cancer (genotype C or Val/Val) may become important for the prevention of lung cancer.     

雌激素相关基因及生殖因素对乳腺癌协同作用

目的研究CYP1A1 MspⅠ，ERα PvuⅡ，ERα XbaⅠ基因多态性与生殖、生育因素在乳腺癌发生中的协同作用。方法本研究样本来自1999年1月-2001年4月，在上海市进行的以人群为基础的乳腺癌病例对照研究。采用PCR-限制性酶切长度多态性法（PIER—RFLP）对282例乳腺癌病例和298例口腔黏膜细胞对照标本进行3个候选基因位点的检测，拟合Logistic回归模型，估计基因多态与生殖、生育因素对乳腺癌协同作用OR值和95％CI，并采用Rothman方法对基因一环境的协同作用进行估计。结果良性乳腺疾病史与CYPIA1 MspⅠ多态存在正相加协同作用。与基准组比较，OR为5．66（95％CI=3．28-9．76），协同作用指数S=2．05，协同作用归因比AP=0．42，经检验差异有统计学意义（P〈0．05）。未发现其他生殖、生育因素与3个基因位点有显著的协同作用。结论良性乳腺疾病史与CYP1A1 MspⅠ基因之间存在增加乳腺危险的协同作用。     

CYP1A1 MspI Polymorphism and Cervical Carcinoma Risk in the Multi-Ethnic Population of Malaysia: a Case-Control Study

Background: Tobacco smoking is considered a risk factor for cervical cancer development due to the presence of tobacco based carcinogenic metabolites in cervical cells of female smokers. In this study, we investigated the role of the T3801C (MspI) polymorphism of CYP1A1, a gene encoding an enzyme necessary for the initiation of tobacco based carcinogen metabolism, on cervical cancer risk. The T to C substitution may alter CYP1A1 activities, potentially elevating cervical cancer risk. Since results of gene-disease association studies vary according to the study population, the multi-ethnic population of Malaysia provides an excellent representative cohort for identifying and comparing the cervical cancer risk among the 3 major ethnics in Southeast Asia in relation to CYP1A1 MspI polymorphism. Materials and Methods: A total of 195 Thin Prep Pap smear samples from HPV negative and cancer free females were randomly selected as controls while 106 formalin fixed paraffin embedded samples from females with invasive cervical cancer were randomly selected for the cases group. The polymorphisms were identified using restriction fragment length polymorphism (RFLP) PCR. Results: We found no significant associations between CYP1A1 MspI polymorphism and cervical cancer in the general Malaysian female population. However, upon ethnic stratification, the variant C/C genotype was significantly associated with a 4.66-fold increase in cervical cancer risk in Malay females (95% CI= 1.21 17.9; p=0.03). No significant association was observed in the Chinese and Indian females. Additionally, there were no significant associations in the dominant model and allele frequency model analysis in both the general and ethnically stratified female population of Malaysia. Conclusions: Our findings suggest that the C/C genotype of CYP1A1 MspI polymorphism is associated with the development of cervical carcinoma in the Malay females of Malaysia.     

DNA polymorphisms in linkage disequilibrium at the 3' end of the human APO AII gene: relationships with lipids, apolipoproteins and coronary heart disease

Two investigations were undertaken to analyze the 3' region of the apolipoprotein AII (Apo AII) gene in patients with myocardial infarction (MI) and controls. Previous studies have suggested that a MspI polymorphism in this gene may be associated with hypertriglyceridaemia, high levels of HDL cholesterol and Apo AII. To verify this hypothesis, the distribution of MspI genotypes and their possible associations with several plasma lipid variables were studied in 882 subjects (411 cases with MI and 471 controls) from the ECTIM study. There were no differences in genotype and allele frequencies between cases and controls, and no differences in lipid variable levels in controls carrying the less frequent MspI allele vs other controls. Using single-strand conformation polymorphism (SSCP) analysis, we detected a new polymorphism which caused by a C-to-T transition located in the third intron near the splice junction site (acceptor). This polymorphism modifies a Bst Nl restriction site. The ECTIM population was screened for this new marker, and no significant associations with MI and plasma lipid levels were found. Our results suggest that these two variants located in the coding region of the Apo AII gene are unlikely to contribute significantly to the level of plasma lipid variables and the risk of coronary heart disease (CHD) in the European population.     

基质金属蛋白酶-2和-9在食管鳞状细胞癌中的过表达

目的 探讨人食管鳞癌组织中基质金属蛋白酶(MMP)-2、-9的表达及其临床意义.方法 采用免疫组织化学法,检测57例食管鳞癌及10例食管正常黏膜石蜡标本中MMP-2、-9蛋白的表达情况.结果 食管鳞癌组织中MMP-2及MMP-9阳性表达率(40.3%,61.4%)显著高于正常组织(0.0%,10.0%)(P＜0.05).食管鳞癌组织中MMP-2及MMP-9阳性表达与淋巴结转移和癌组织浸润深度有显著相关性(P＜0.05),而与患者的性别、年龄和组织分化程度无显著相关性(P＞0.05).结论 MMP-2和MMP-9在食管癌中显著高表达,与食管癌的转移及侵袭有关,其异常表达可能共同参与食管癌的发生、发展过程,检测MMP-2、-9可作为食管癌病理学特点的参考指标.     

载脂蛋白AⅠ基因启动子区G→A置换及内含子ⅠC→T置换与冠心病的关联

目的探讨冠心病患者载脂蛋白(apo)AⅠ基因MspⅠ酶切位点变异频率及其对血脂水平的影响.方法应用多聚酶链反应对成都地区汉族178例正常人和118例冠心病患者apoAⅠ基因启动子(-78bp)及内含子Ⅰ(+83bp), MspⅠ限制性片段多态性进行分析.结果冠心病组及对照组apoAⅠ基因MspⅠ的多态性以G/G基因型占优势.apoAⅠ基因启动子区MspⅠ酶切位点A等位基因频率冠心病组与对照组比较有升高趋势,但无显著性差异(0.331 vs. 0.270,P=0.067),+83bp T等位基因频率冠心病组较对照组显著升高(0.072 vs. 0.034,P<0.05).在所研究对象中具有A/A基因型者血清TG水平较具有G/G基因型者显著升高(P<0.05).结论 apoAⅠ基因内含子Ⅰ(+83bp)MspⅠ酶切位点的突变与中国人冠心病有一定关联,apoAⅠ基因A/A基因型对血清甘油三酯的升高有一定影响.     

New MspI polymorphism at +83 bp of the human apolipoprotein al gene: Association with increased circulating high density lipoprotein cholesterol levels

We recently showed that loss of a MspI restriction site in the 5′-end (intron 1) of the apolipoprotein (apo) AI gene is due to a C to T transition (+83 bp) and/or a G to A transition (+84 bp). Since this region may be relevant to the regulation of apo AI gene expression and therefore to plasma high density lipoprotein cholesterol (HDL-C), we explored the association between this MspI polymorphic site and circulating HDL-C levels in 243 healthy Caucasians. There were 143 aged 18–67 years (60 males and 83 females) and 100 aged 6–12 years (58 males and 42 females). We also compared this association with a known MspI polymorphic site, a G to A transition at −75 bp of the apo AI gene. The rare allele (−) frequency for the polymorphism at +83 bp was 4.1% and 22.1% for the polymorphism at −75 bp. Subjects heterozygous for the loss of the MspI restriction site at +83 bp (genotype: M2+−, n = 20) had higher HDL-C levels than M2++ subjects (mean ± SD: 1.73 ± 0.31 vs. 1.41 ± 0.39 mmol/l, P < 0.05 for adults; 1.71 ± 0.33 vs. 1.34 ± 0.29 mmol/l, P < 0.01 for children). Adults with the G to A substitution at −75 bp also had higher HDL-C levels (1.56 ± 0.36 mmol/l for AA, 1.53 ± 0.38 mmol/l for GA, and 1.36 ± 0.38 mmol/l for GG, P < 0.05); this difference was not observed in the children. The MspI polymorphisms at both sites were in linkage disequilibrium. Their joint effect on the HDL-C levels was also significant and individuals with rare alleles (−) at both sites had the highest HDL-C levels. In an analysis of variance, the MspI polymorphism at +83 bp, and at −75 bp and gender independently accounted for 6.5%, 1.7%, and 5.9%, respectively, of the variance in circulating HDL-C levels when age was controlled as a covariate. We conclude that loss of the MspI site by the C to T (+83 bp) and/or the G to A (+84 bp) transitions is highly associated with increased HDL-C levels. The association appears to be more significant than that of the G to A transition at −75 bp. © 1996 Wiley-Liss, Inc.