内皮素受体B基因多态性与慢性阻塞性肺疾病及吸烟因素的相关性研究

目的探讨内皮素受体B（EDNRB）基因的第4外显子-30G／A处单核苷酸多态性与慢性阻塞性肺疾病（COPD）易感性及吸烟因素之间关系。方法采用PCR—RFLP技术,检测并分析EDNRB基因一30G／A（L277L）单核苷酸多态性位点在COPD组和健康对照组中的基因型频率、等位基因频率,同时分析该多态性位点与吸烟相关COPD之间是否存在相关性。结果研究发现EDNRB基因一30G／A处基因型频率分布在COPD组和对照组之间差异有统计学意义（P〈0．05）,但等位基因频率的分布在两组之间差异无统计学意义（P〉0．05）。在对COPD组和对照组中吸烟者的基因型分布频率和等位基因分布频率比较后,发现两组之间差异无统计学意义（P〉0．05）,提示该突变位点与吸烟所致的COPD之间可能无相关性存在。结论EDNRB一30G／A位点多态性可能与COPD易感性相关,与吸烟相关的COPD无关。     

家族性高胆固醇血症患者低密度脂蛋白受体基因新的突变类型一例

目的：分析一例家庭性高胆固醇血症患的低密度脂蛋白受体基因突变位点。方法：以患儿的基因组DNA为模板，用聚合酶链反应（PCR）扩增该基因的18个外显子。用单链构象多态性（SSCP）方法分析检测PCR产物，对电泳结果异常进行DNA测序。结果：单链构象多态性分析发现患儿第10外显子存在一异常条带。DNA测序证实患儿第10外显子发生N515S纯合错义突变。结论：该病例为一个新的LDLR突变位点；聚合酶链反应－单链构象多态性分析（PCR－SSCP）可用于该突变位点的诊断。     

细胞毒性T淋巴细胞相关抗原4/CT60基因多态性与溃疡性结肠炎的关系

目的探讨细胞毒性T淋巴细胞相关抗原4（CTLA4）CT00位点基因多态性与中国湖北地区汉族人溃疡性结肠炎（UC）的相关性。方法共收集64例UC患者和109例正常对照静脉血标本提取基因组DNA。采用多聚酶链反应．限制性片段长度多态性分析（PCR-RFLP）方法检测CT60G／A（rs3087243）位点的多态性，计算CTLA4的基因型和等位基因频率。结果CT60G／A基因型分布及等位基因频率，UC组与正常对照组比较差异无显著性，中国湖北地区汉族人群CTLA4CT60基因型和等位基因分布与日本人群相似。而与西班牙人群分布存在显著差异。结论CT60基因多态性与中国湖北汉族人UC易感性无关。     

2型糖尿病与簇蛋白基因关联的初步研究

目的探讨2型糖尿病（T2DM）与簇蛋白（Glu）基因多态性的关联。方法聚合酶链式反应-变性梯度凝胶电泳（PCR-DGGE）和DNA测序技术分析62例T2DM患者及60例正常对照者的Glu基因外显子2和外显子5基因多态性。结果Glu基因外显子2发现1个多态位点：1963（＋A）;外显子5发现3个多态位点：（1）7476（-A）,（2）7534（-C）,（3）7521C→T。结论Glu基因外显子2和外显子5多态性在2型糖尿病组和正常对照组多态性位点基因频率分布差异有统计学意义。因此,Clu基因外显子2和外显子5多态性可能与T2DM易感性有关联。     

半胱氨酸蛋白酶抑制因子Cystatin C基因与中国人脑卒中的关系

目的本文通过多中心病例．对照研究，拟探讨Cystatin C基因启动子区多态性位点rs2897119（-177C／T）和rs6114208（-1704C／G）与中国人脑卒中和血浆中半胱氨酸水平的关系。方法采用聚合酶链反应-限制性片段长度多态性（PCR-RFLP）分析技术，检测脑卒中组520例（血栓性脑梗塞组211例，腔隙性脑梗塞组145例，脑出血组164例），和非脑卒中对照组630例Cystatin C基因多态性的分布。用高效液相色谱仪和荧光检测仪测定血浆总Hey水平。结果脑卒中组和对照组Cystatin C基因rs2897119和rs6114208多态性基因型频数分布符合Hardy-Weinber平衡。两组rs2897119和rs6114208多态性基因型和等位基因型分布差异无明显统计学意义。各亚型组与对照组的基因型和等位基因型相比差异也均无显著统计学意义。两组多态性基因型与相应血浆同型半胱氨酸的水平也无显著统计学意义。结论Cystatin C基因多肽位点rs2897119（-177C／T）和rs6114208（-1704C／G）与中国人脑卒中和血浆同型半胱氨酸的水平不存在关联关系。Cystatin C基因多肽位点rs2897119和rs6114208可能不是中国人脑卒中的遗传影响因素。     

冠心病患者对氧磷酶启动子基因-108C/T、-162A/G多态性分析

目的探讨中国人对氧磷酶（PON1）基因-108C／T、-162A／G遗传多态性与冠心病（CHD）的关系。方法应用多聚酶链反应（PCR）对86例冠心病患者和128例正常对照者PON1基因酶切位点的限制性片段长度多态性（RFLP）进行研究。结果PON1—108C／T多态性位点基因型分布在CHD组和对照组间存在显著性差异（P〈0．05）。PON1-162A／G多态性位点基因型频率和等位基因频率在两组间未见显著性差异。CHD不随两个多态性位点危险基因型个数的增加而发病率呈上升趋势。结论PON1—108C／T基因多态性与中国人群CHD相关联，PON1—108C／T和PON1—162A／G基因多态性对CHD的影响无协同性。     

OCTN1/OCTN2基因多态性与中国汉族人群炎症性肠病相关性的初步研究

近年研究证实OCTN1/OCTN2基因单核苷酸多态性（SNP）与西方白种人群克罗恩病（CD）明显相关,其中OCTN1基因rs1050152位点和OCTN2基因rs2631367位点SNP与CD的相关性尤为显著.目的：初步探讨OCTN1基因rs1050152位点和OCTN2基因rs2631367位点基因多态性与中国汉族人群炎症性肠病（IBD）的相关性.方法：KR联合直接测序法检测45例CD患者、45例溃疡性结肠炎（UC）患者和50名正常对照者OCTN1基因rs1050152位点和OCTN2基因rs2631367位点的基因型,比较各组基因型和等位基因频率.结果：CD,UC和正常对照组间OCTN1基因rs1050152位点基因型和等位基因频率差异均无统计学意义（R〉0.05）;三组OCTN2基因rs2631367位点均未发现突变基因型.结论：OCTN1基因rs1050152位点和OCTN2基因rs2631367位点基因多态性与中国汉族人群IBD不相关.     

海南黎族冠心病患者胆固醇酯转运蛋白基因多态性测定及分析

目的探讨胆固醇酯转运蛋白（CETP）基因多态性在海南黎族人群冠心病发病中的作用。方法应用等位基因特异性PCR方法,对海南黎族47例冠心病患者（观察组）及330例健康查体者（对照组）的CETP基因6种多态位点进行检测,比较两组基因型频率和等位基因频率。结果两组TaqIB突变位点可检测出B1B1、B1B2、B2B23种基因型,I405V突变位点可检测出Ⅱ、Ⅳ、ⅤⅤ3种基因型,D442G突变位点可检测出DD、DG2种基因型;两组TaqIB突变位点的等位基因频率及D442G突变位点的基因型分布和等位基因频率存在显著差异（P〈0.05）。结论 CETP基因TaqIB和D442G位点的多态性与海南黎族人群冠心病显著相关;本研究样本数较少,有关CETP基因多态性与海南黎族人群冠心病的确切关系尚有待于开展大样本流行病学调查。     

内皮细胞型NO合酶基因第7外显子894G→T点突变与糖尿病肾病相关性研究

目的 探讨内皮细胞型NO合酶（eNOS)基因第7外显子894G→T点突变与中国北方汉族人2型糖尿病（T2DM）合并肾病（DN）之间的关系。方法 运用聚合酶链式反应限制性片段长度多态性技术（PCR－RFLP），结合DNA测序技术，检测了228例中国北方汉族人的eNOS基因第7外显子894G→T错义突变位点的基因型，其中T2DM患者143例（DN79例），健康成人85例，并对各组间的等位基因频率与基因型频率进行了比较。结果 ①T2DM组的T等位基因及TG基因型频率与正常对照（N）组无显著性差异（P＞0．05）。②DN＋组T等位基因及TG基因型频率显著高于糖尿病非肾病患者（P＜0．05）。③SBP、HbA1c、TC、TG和eNOS基因第7外显子894G→T点突变均与糖尿病肾病有关（P＜0．05）。结论 eNOS基因第7外显子894G→T点突变的T等位基因可能是中国人2型糖尿病易患肾病的独立危险因素。     

原发性肝癌患者DNA损伤修复酶基因多态性分析

目的探讨DNA损伤修复酶基因多态性与原发性肝癌发生发展的关系。方法 PCR-RFLP法检测150例肝癌患者（肝癌组）和150例健康体检者（对照组）DNA损伤修复酶基因的表型,并进行比较。结果肝癌组XRCC1 Arg399Gln位点野生型的比率明显低于对照组（P〈0.05）;XRCC1（Arg194Trp、Arg280His）、hOGG1Ser326Cys位点多态性型别在两组中出现的频率相近（P〉0.05）;各基因位点不同表型者的尿8-羟基脱氧鸟苷（8-OHdG）水平相比,P均〉0.05。结论 DNA修复酶基因多态性与肝癌的发生发展无明确的相关关系。     

Clinical relationship between EDN-3 gene, EDNRB gene and Hirschsprung＇s disease

AIM: To investigate the mutation of EDNRB gene and EDN-3 gene in sporadic Hirschsprung‘s disease (HD) in Chinese population. METHODS: Genomic DNA was extracted from bowel tissues of 34 unrelated HD patients which were removed by surgery. Exon 3, 4, 6 of EDNRB gene and Exon 1, 2 of EDN-3 gene were amplified by polymerase chain reaction (PCR) and analyzed by single strand conformation polymorphism (SSCP). RESULTS: EDNRB mutations were detected in 2 of the 13 short-segment HD. One mutant was in the exon 3, the other was in the exon 6. EDN-3 mutation was detected in one of the 13 short-segment HD and in the exon 2. Both EDNRB and EDN-3 mutations were detected in one short-segment HD. No mutations were detected in the ordinary or longsegment HD. CONCLUSION: The mutations of EDNRB gene and EDN-3 gene are found in the short-segment HD of sporadic Hirschsprung‘s disease in Chinese population, which suggests that the EDNRB gene and EDN-3 gene play important roles in the pathogenesis of HD.     

MEN1基因突变所致多内分泌腺瘤病1型二例及其家系研究

目的 探讨多内分泌腺瘤病1型的发病与MEN1基因突变的相互关系及其遗传特征。方法 从2个家系中的患者及其家庭成员抽提外周血淋巴细胞基因组DNA，运用PCR、DNA测序分析技术检测MEN1基因所有10个外显子，进一步用亚克隆测序鉴定其杂合性。结果发现两个家系中的先证者MENI基因均在第二号外显子存在杂合突变，分别为372-373insACCTGTCTATCATCGCCG和357-360delCTGT，前一种突变为一种新的突变类型。结论 在2个多内分泌腺瘤病1型中国人家系检出2种MEN1基因突变，其中MEN1基因372-373insACCTGTCTATCATCGCCG为一种新的MEN1基因突变。     

Analysis of NPM1 Gene Mutations in Chinese Adults with Acute Myeloid Leukemia

Many European groups have recently described that mutations at exon-12 of the nucleophosmin (NPM1) gene are the most frequent genetic lesion in patients with acute myeloid leukemia (AML), especially in the presence of a normal karyotype. This study explored the prevalence and clinical profile of NPM1 mutations in a cohort of 156 Chinese adults with AML. NPM1 exon-12 mutations were detected using direct sequencing or fragment analysis of genomic DNA polymerase chain reaction products. NPM1 mutations were present in 28.2% of the overall population, including 1/1 (100%) of M0, 11/27 (40.7%) of M1, 11/46 (23.9%) of M2, 0/29 (0%) of M3, 2/9 (22.2%) of M4, 18/39 (46.2%) of M5, and 1/5 (20.0%) of M6. NPM1 gene mutations were more prevalent in patients with a normal karyotype (37 of 90; 41.1%) when compared with patients with karyotypic abnormalities (7 of 66; 10.6%;P < .001). Sequence analysis of 25 NPM1-mutated cases revealed known mutations (type A, D, N(M), and P(M)) as well as one novel sequence variation (here named as type S). All mutational types were heterozygous and showed a 4 bp insertion. NPM1 mutations were significantly associated with old age (P < .05), high peripheral white blood cell count (P < .05), and the subtypes of French-American-British categories M1/M5, but negatively associated with expression of CD34 (P < .05) and CD117 (P < .05). Thus, this study provides the methods of NPM1 exon-12 mutations detection and related clinical data of NPM1 mutated cases in a Chinese population.     

变性梯度凝胶电泳分析检测结直肠癌APC和p53基因突变

目的: 探讨APC和p53基因突变在结直肠癌中的意义.方法:采用变性梯度凝胶电泳(DGGE), DNA测序法分析15例正常人和60例散发性结直肠癌标本的APC基因15外显子和p53基因第5, 7外显子的基因突变.结果: 在结直肠癌组检出14例APC和16例p53基因突变, 测序证实其中13/14例APC发生在突变集中区域;9/16例p53基因突变位于第5外显子, 7/16例p53基因突变位于第7外显子, 2例同时检出了APC基因和p53基因突变.结论:DGGE是一种快速、简便、高效、灵敏的突变检测技术. 同时也证明APC基因突变和p53基因突变均参与了结直肠癌的发生、发展过程.     

广西艾滋病疫情严重地区壮族和汉族普通人群CCR5 CCR2 SDF1等位基因检测及其意义

目的调查艾滋病病毒Ⅰ型（HIV—1）感染相关的CCR5、CCR2及SDF1等位基因,在广西壮族和汉族普通人群中的多态性及其流行病学意义。方法在广西天等县和南宁市,选取150名壮族和90名汉族健康人作为研究对象,采集外周血,提取基因组DNA,用聚合酶链反应（PCR）扩增CCR5、CCR2及SDF1基因的特定片段。直接根据PCR扩增结果分析CCR5基因多态性,运用限制性片段长度多态性（RFLP）技术分析CCR2及SDF1的多态性。结果全部研究对象的CCR5基因均为野生型,未发现CCR5△32突变;CCR2—64Ⅰ等位基因频率在壮、汉族普通人群中分别为25．7％、26．1％;SDF1—3’A等位基因频率分别为27．7％和27．2％。结论广西壮、汉族普通人群缺乏艾滋病抗性的CCR5△32等位基因,而CCR2-641和SDF1—3’A等位基因频率较高。该研究为深入研究广西壮、汉族普通人群对于HIV—1感染的遗传易感性,以及对艾滋病疫情和病程的影响提供了比较全面、可靠的数据。     

Point mutation of 5＇ noncoding region of BCL-6 gene in primary gastric lymphomas

AIM: To investigate the mutations of the 5' noncoding region of BCL-6 gene in Chinese patients with primary gastric lymphomas. METHODS: PCR and direct DNA sequencing were used to identify BCL-6 gene mutations in the 5' noncoding region in 29 cases of gastric diffuse large B-cell lymphoma (DLBCL) and 18 cases of gastric mucosa-associated lymphoid tissue (MALT) lymphoma as well as 10 cases of reactive hyperplasia of lymph node (LRH). RESULTS: Six of 29 gastric DLBCLs (20.7%), 4 of 18 gastric MALT lymphomas (22.2%) and 1 of 10 LRHs(10%) were found to have mutations. All mutations were single-base substitutions and the frequency of single-base changes was 0.20×1O~(-2)-1.02×1O~(-2)per bp. CONCLUSION: Point mutations in the 5' noncoding region of BCL-6 gene are found in Chinese patients with primary gastric DLBCLs and MALT lymphomas, suggesting that they may, in some extent, participate in the pathogenesis of primary gastric DLBCLs and MALT lymphomas.     

ABCB11 gene mutations in Chinese children with progressive intrahepatic cholestasis and low γ glutamyltransferase

Background: Progressive familial intrahepatic cholestasis type 2 (PFIC2) is a severe autosomal recessive liver disorder of childhood that can cause cholestasis and progress to end‐stage liver disease. ABCB11 gene mutations causing PFIC2 have been reported in some population groups, but not in mainland Chinese. Aims: To elucidate the existence of and characterize ABCB11 gene mutations in mainland Chinese with progressive intrahepatic cholestasis and low γ glutamyltransferase (GGT). Methods: Twenty‐four children presenting with progressive intrahepatic cholestasis and low GGT were admitted to a tertiary paediatric hospital in eastern China from January 2004 to July 2007. All encoding exons and flanking areas of the ABCB11 gene were sequenced. Hepatic histopathology results were obtained by review of the medical record. Results: Twelve novel mutations of ABCB11 gene were found in seven patients: three nonsense mutations, six missense mutations, two splicing mutations and one intronic mutation. Giant cell transformation of hepatocytes was demonstrated in all the four patients with ABCB11 mutations and four of 12 patients without mutations in coding sequences of ABCB11 gene who received liver needle biopsy. Conclusions: ABCB11 gene mutations play an important role in Chinese patients with progressive intrahepatic cholestasis and low GGT. The characteristics of ABCB11 gene mutations in Chinese are different from other population groups. Histological examination may be helpful in diagnosis of PFIC2.     

Islet autoimmunity and genetic mutations in Chinese subjects initially thought to have Type 1B diabetes.

AIMS: To explore the contribution of islet autoimmunity and genetic mutations in Chinese patients initially thought to have Type 1B diabetes. METHODS: A group of 33 Chinese patients with newly diagnosed Type 1B diabetes, were identified by the absence of autoantibodies to glutamic acid decarboxylase (GAD), IA-2, insulin, thyroid globulin or thyroid peroxidase, or high-risk HLA-DQ haplotypes. The cohort was further characterized by measurement of autoantibodies to carboxypeptidase H (CPH) and SOX13 using radioligand assays, and testing for genetic mutations associated with MODY3/MODY6 and mitochondrial diabetes. Mutations of HNF-1alpha (MODY3) and neuroD1/beta2 (MODY6) genes were screened using the single-strand conformation polymorphism (SSCP) technique and sequencing. Mitochondrial DNA mutations were analysed with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: Within the cohort, we found one patient with a novel mutation, R321H (CGC-->CAC) in exon 5 of the HNF-1alpha gene, one with ND1 mt3316 G-->A mutation in mitochondrial DNA, five with Ala45Thr polymorphisms in the neuroD1/beta2 gene, and two patients with autoantibodies to SOX13. CONCLUSIONS: Some of the Chinese patients originally thought to have Type 1B diabetes do have other evidence of islet autoimmunity and genetic mutations involved in the underlying aetiology. This suggests that more rigorous screening for these conditions is needed before classifying subjects as having Type 1B diabetes.     

青壮年猝死综合征家系及散发病例SCN5A基因突变的检测

目的研究中国人青壮年猝死综合征（SUNDS）病例是否存在SCN5A基因突变。方法应用直接测序技术对74例散发病例及两个家系病例的4个成员的血纱样本进行SCN5A基因突变的检测。结果在散发病例发现突变位点R1512W、A1121A,其中A1121A为新发现的突变。结论中国人SUNDS病例存在SCN5A基因突变,后者可能与部分SUNDS的发生相关。