胰岛素受体底物1基因Gly972Arg多态性与海南黎族2型糖尿病的关系

目的 研究胰岛素受体底物1(IRS-1)基因Gly972Arg突变与海南黎族2型糖尿病(T2DM)的关系.方法应用聚合酶链反应-限制性片段长度多态性技术,对78例海南黎族T2DM患者(T2DM组)和78例糖耐量正常人群(对照组)进行IRS-1基因Gly972Arg突变位点基因型检测.结果 对照组IRS-1基因Gly972Arg突变频率5%,T2DM组未发现该基因突变.结论 IRS-1基因Cly972Arg突变可能不是海南黎族T2DM的重要遗传因素.     

低钾性周期性麻痹的分子生物学基础

低钾性周期性麻痹是一种与遗传有关的离子通道疾病,目前所知的突变分别是L型电压门控Ca2+通道编码基因(CACNAlS)Arg528His、Arg1086Cys、Arg1239Gly、Arg1239His、电压门控Na+通道编码基因(SCN4A)Arg669His、Arg672Gly、Arg672His、Arg672Ser、Arg672Cys、Pro1158Ser及电压门控K+通道编码基因(KCNE3)Arg83His.不同的突变类型具有不同的临床特点,CACNAlS Arg528His和CACNAIS Arg1239His突变者发病较早;SCN4A Arg672Gly突变者发病更早,在女性中表现为不完全外显,发作后肌痛重,病理表现为管性聚集,乙酰唑胺治疗会致肌无力的加重.突变引起的氨基酸改变影响了通道电压感受器的功能,产生异常的静息膜电位,引起兴奋收缩偶联的改变从而导致肌无力的发生.     

Ⅰ型原发性草酸尿症家系AGT基因新突变鉴定

目的分析Ⅰ型原发性草酸尿症(PH)家系的遗传方式、临床特征及致病基因丙氨酸乙醛酸氨基转移酶(AGT)突变。方法以2012年11月上海交通大学医学院收治的1例PH患者为研究对象,通过家系调查和血尿草酸浓度离子色谱分析,直接测序法分析AGT基因。同时选取100名健康正常人作为对照。结果该家系共有2例患者确诊为Ⅰ型原发性草酸尿症(PH1),遗传特点符合复杂常染色体隐性遗传。AGT基因测序分析发现先证者与其父亲8号外显子均存在一个新的杂合插入突变(C.946_947insAG),在蛋白水平上该突变引起AGT第275位丝氨酸开始的移码突变,导致羧基末端的119个氨基酸替换缩短为异常的38个氨基酸(P.Ser275Arg fs*38),且先证者与其母亲第2号外显子第467位碱基均存在杂合点突变(C.467GA),导致(P.Gly116Arg),此突变为已报道过的致病点突变。结论一个新的AGT突变位点被确定与隐性遗传性PH1有关。这是国内首次对遗传性PH1进行基因学研究。     

肥厚型心肌病致病基因型与临床表现的关系及基因筛查在肥厚型心肌病筛查及疾病鉴别诊断中的作用

目的:分析肥厚型心肌病(HCM)致病基因型与临床表现的关系及基因筛查在HCM筛查及疾病鉴别诊断中的作用。方法:选择一个HCM家系共14人,多重靶向测序技术对先证者的26个已知最常见的HCM致病基因进行全外显子捕获测序,用Sanger测序对发现的突变进行验证并对其他家系成员及307名健康对照进行该突变位点的筛查,分析其基因型与临床表型的特点。结果:先证者及其子携带MYH7基因c.2146 GA(Gly716Arg)突变,该突变位于MYH7基因19号外显子,导致第716位氨基酸残基由Gly变为Arg,其他25个基因未发现突变。Sanger测序验证后对其他家系成员进行突变筛查,其他家庭成员及对照组未发现该突变,该突变与HCM在该家系中共分离。先证者携带的致病突变为从头突变,并遗传给其子。先证者临床表现为发病早(14岁)、劳力性呼吸困难、胸痛、心悸、心力衰竭,其子出生时即发现心肌肥厚。其父虽然室间隔肥厚(15 mm),但结合其年轻时曾为运动员的经历及遗传筛查的阴性结果,可基本排除其为HCM患者,考虑为生理性肥厚。结论:该家系HCM由MYH7基因从头突变p.Gly716Arg导致,该突变临床发病早,症状较重,预后较差,为恶性突变。基因筛查在HCM家系筛查及疾病鉴别诊断中有重要意义。     

低钾性周期性麻痹的分子生物学基础

低钾性周期性麻痹是一种与遗传有关的离子通道疾病，目前所知的突变分别是L型电压门控Ca^2 通道编码基因(C4CNIS)Arg528His、Arg1086Cys、Arg1239G1y、Arg1239His、电压门控Na’通道编码基因(SCN4A)Arg669His、Arg672G1y、Arg672His、Arg672Ser、Arg672Cys、Pro1158Ser及电压门控K^ 通道编码基因(KCAE3)Arg83His。不同的突变类型具有不同的临床特点，CACNAIS Arg528His和CAC-NAIS Arg1239His突变者发病较早；SCN4A Arg672Gly突变者发病更早，在女性中表现为不完全外显，发作后肌痛重，病理表现为管性聚集，乙酰唑胺治疗会致肌无力的加重。突变引起的氨基酸改变影响了通道电压感受器的功能，产生异常的静息膜电位，引起兴奋收缩偶联的改变从而导致肌无力的发生。     

β2肾上腺素受体Arg16Gly基因多态性与原发性高血压的相关性研究

目的:研究河北地区汉族人β2肾上腺素受体(β2-Adrenergic Receptor, β2-AR)Arg16Gly基因多态性与原发性高血压(EH)的关系.方法:选择514例EH患者及550例正常人,采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)分析β2-AR Arg16Gly基因型.结果:EH组β2-AR Arg16Arg,Arg16Gly,Gly16Gly基因型频率分别为21.3%、54.8%和23.9%,正常对照组分别为28.2%、68.2%和3.6%,2组间3种基因型频率差异有统计学意义(P<0.05).EH组β2-AR Arg16等位基因频率为51.8%,Gly16等位基因频率为48.2%;正常对照组分别为63.9%和36.1%,2组间等位基因频率比较,差异有统计学意义(P<0.05).结论:β2-AR Arg16Gly多态性Gly等位基因可能是EH的遗传易感基因.     

新疆地区维吾尔族先天性心脏病患者CITED2基因突变分析

【】 目的：研究新疆维吾尔族先天性心脏病(congenital heart disease,CHD)患者与CITED2基因突变的关系。方法：收集150例散发型维吾尔族CHD患者和150例健康维吾尔族人群血液样本进行DNA提取、目的基因聚合酶链反应及测序,并与GeneBank进行比较以识别基因突变,并分析氨基酸序列。结果：在1例室间隔缺损患者发现1个新的纯合突变c.574A>G,并导致相应氨基酸序列错义突变(p.Ser192Gly),该突变在CITED2基因丝氨酸-甘氨酸富含区,而在健康对照组中未发现此突变。结论：新疆维吾尔族CHD患者中首次发现了CITED2基因纯合突变,Serl92GIy突变所在序列呈现一定保守性,可能与CHD发生有关。     

中国汉族家族性肥厚型心肌病人群MYH7基因Arg723Gly突变分析

目的研究中国汉族人群家族性肥厚型心肌病的致病基因突变位点,分析基因型与表型的关系。方法对5个肥厚型心肌病家系的先证者进行β-肌球蛋白重链基因扫描,聚合酶链反应扩增其功能区的外显子片段,双脱氧末段终止法测序。对阳性结果患者进行家系调查,收集临床资料,分析其临床表型。结果在1个家系中发现Arg723Gly杂合突变,而正常对照组同一位置未见异常,此为我国患者中首次发现Arg723Gly突变。结论β-肌球蛋白重链可能是我国家族性肥厚型心肌病的常见致病基因之一。Arg723Gly所致肥厚型心肌病外显率高、临床症状出现较早、进展较快、易发生心力衰竭、预后较差,心脏室、房扩大也较常见,是一种恶性突变。     

β1肾上腺素能受体Gly 389Arg多态性与原发性高血压的关系

目的:探讨汉族人群β1肾上腺素能受体Gly 389Arg多态性与原发性高血压的关系. 方法:采用聚合酶链反应-限制性长度片段多态性技术分析原发性高血压患者和正常人群β1肾上腺素能受体Gly 389Arg多态性.结果:高血压组Arg/Arg,Arg/Gly,Gly/Gly基因型频率分别为59.06%、35.09%、5.85%,正常对照组分别为43.55%、45.97%、10.48%;两组间三种基因型频率分布有统计学差异(x2=7.420,P＜0.05);高血压组Arg等位基因频率为76.61%,Gly等位基因频率为23.39%,正常对照组分别为66.53%、33.47%,两组间等位基因频率分布存在统计学差异(x2=7.299,P＜0.05);高血压组Arg纯合子基因型频率和Arg等位基因频率均明显高于对照组. 结论:β1肾上腺素能受体Gly 389Arg多态性可能与原发性高血压发病有关.     

海南汉族2型糖尿病患者胰岛素受体底物1基因Gly972Arg多态性的研究

目的 研究胰岛素受体底物1（IRS。1）基因Gly972Argr矢变与海南汉族2型糖尿病（T2DM）的关系。方法应用聚合酶链反应-限制性片段长度多态性技术对海南汉族60例T2DM患者和60例糖耐量正常人群进行IRS-1基因Gly972Arg突变位点基因型检测。结果T2DM组IRS-1基因Gly972Arg突变频率13．3％,对照组IRS-1基因Gly972Arg突变频率3．3％,两者比较有统计学差异（r=7．2,P〈0．01）。结论IRS—1基因Gly972Arg突变可能与海南汉族T2DM的发生有相关性。     

胰岛素受体底物1（IRS-1）基因Gly972Arg突变与中国人糖尿病的相关性

目的 研究胰岛素受体物１（ＩＲＳ－１）基因Ｇｌｙ９７２Ａｒｇ突变与中国人糖尿病的相关性。方法 随机选取上海地区中国人３５９例,用ＰＣＲ／ＢｓｔＮ１酶解法检测Ｇｌｙ９７２Ａｒｇ突变。结果 本组中国人中仅发现２例该突变且均为非糖尿病者。结论 该突变可能不是中国人糖尿病的重要遗传因素。     

Molecular scanning for mutations in the insulin receptor substrate-1 (IRS-1) gene in Turkish with type 2 diabetes mellitus

Insulin receptor substrate-1 (IRS-1) is an endogenous substrate for the insulin receptor tyrosine kinase, which plays a key role in insulin signaling. Recent studies have identified several polymorphisms in the human IRS-1 gene (Irs-1) that are increased in prevalence among type 2 diabetic patients. To determine whether variation in the Irs-1 contributes to genetic susceptibility to type 2 diabetes in Turkish people, PCR-RFLP and DNA sequencing method were utilized to analyze the coding region of Irs-1 in 70 subject and 116 control patients. Three missense mutations were detected (Gly972Arg, Ala512Pro, Ser892Gly). There was no significant association found with any of these variants and diabetes. The Gly972Arg mutation, however, was relatively more common in with 10/70 diabetic patients and 15/116 non-diabetic controls being heterozygous and 1/70 being and 0/116 non-diabetic controls being homozygous for this variant. As a conclusion, Ala512Pro, Ser892Gly mutations were rare and Met613Val, Ser1043Tyr and Cys1095Tyr mutations were not found in the populations studied. Gly972Arg is more common than other known mutations in our population but may not be a major determinant in genetic susceptibility to type 2 diabetes.     

Correlation of mutational analysis to clinical features in Taiwanese patients with Gilbert's syndrome

OBJECTIVES: Mutations in the promoter as well as in the coding region of the bilirubin UDP-glucuronosyltransferase gene (UGT1A1) have been found to be associated with Gilbert's syndrome. However, the genetic basis of Gilbert's syndrome in our population and correlation of these mutations to fasting serum bilirubin levels in patients with Gilbert's syndrome remain to be clarified. METHODS: We applied polymerase chain reaction-based direct-sequencing assays to examine mutations in UGT1A1 gene in 20 unrelated Gilbert's patients and in a family with Gilbert's syndrome. RESULTS: We studied three mutations that were previously reported to be associated with Gilbert's syndrome (i.e., the TATAA-box mutation, Gly71Arg, and Pro229Gln) in 20 patients with Gilbert's syndrome. Of the patients, 16, five, and six were found to have the TATAA-box, Gly71Arg and Pro229Gln mutations, respectively. Seven patients had simultaneous mutations both in the TATAA box and in the coding region. Of note, all six patients with Pro229Gln also had the TATAA-box mutation. Localization of Pro229Gln on the allele containing the TATAA-box mutation was demonstrated in a family with Gilbert's syndrome. The patients simultaneously heterozygous for both the TATAA-box mutation and Gly71Arg usually had serum bilirubin levels similar to those found in the patients homozygous for the TATAA-box mutation, but usually higher than those found in the patients heterozygous for the TATAA-box mutation alone. On the other hand, concurrence of Pro229Gln in patients with TATAA-box mutation or with Gly71Arg did not significantly affect serum bilirubin levels. CONCLUSIONS: The TATAA-box mutation and Gly71Arg are the major causes for Gilbert's syndrome in our population. Concurrence of mutations of Gly71Arg and TATAA-box usually exerts a synergistic effect on hyperbilirubinemia. Pro229Gln, which is regularly linked to the TATAA-box mutation, may not have a significant effect on serum bilirubin levels.     

Intermittent jaundice in patients with acute leukaemia: A common mutation of the bilirubin uridine-diphosphate glucuronosyltransferase gene among Asians

The Gly71Arg mutation of the hepatic bilirubin UDP glucuronosyltransferase (B-UGT) gene associated with Gilbert syndrome prevails among Japanese and its gene frequency is 0.13. Among 20 patients with acute leukaemia, 4 patients showed intermittent unconjugated hyperbilirubinaemia during the course of combined chemotherapy. The Gly71Arg mutation was detected in all 4 patients with hyperbilirubinaemia, but was not found in 16 patients without hyperbilirubinaemia. Two of them were heterozygotes and one was a homozygote for the Gly71Arg mutation, and the other was a compound heterozygote of the Gly71Arg mutation and TA insertion mutation in the TATA box of the B-UGT gene. In addition to the complications leading to hyperbilirubinaemia, including liver damage due to drugs, viral infections or tumour cell infiltrations and alloimmune haemolysis, carrier status for the Gly71Arg mutation should be considered in a patient with leukaemia showing intermittent hyperbilirubinaemia during the course of chemotherapy, especially among Japanese, Koreans and Chinese owing to its prevalence in those populations.     

Association between beta2-adrenergic receptor genetic polymorphisms and nocturnal asthmatic patients of Chinese Han nationality

BACKGROUND: As a result of the finding that the mutation of Arg into Gly at beta(2)-adrenergic receptor (beta(2)-AR)16 loci could promote the downregulation effect triggered by the beta(2)-agonist, it was supposed that Gly16 might be associated with the downregulation of beta(2)-AR in patients with nocturnal asthma. OBJECTIVE: It was the aim of this study to analyze the association between beta(2)-AR genetic polymorphisms and nocturnal asthmatic patients of Chinese Han nationality. METHODS: A polymerase chain reaction allele-specific oligonucleotide hybridization assay was used to determine 16 and 27 loci alleles of beta(2)-AR genetic polymorphisms in 25 nocturnal asthmatic patients (nocturnal asthma group), 22 non-nocturnal asthmatic patients (non-nocturnal asthma group), and 72 healthy people (control group). All people investigated were of Chinese Han nationality. RESULTS: The distribution frequency of genotype Arg/Arg, Arg/Gly, and Gly/Gly at beta(2)-AR 16 loci was 12, 16 and 72% in the nocturnal asthma group; and 27, 41 and 32% in the non-nocturnal asthma group. There was a significant increase in the frequency of genotype Gly/Gly and allele Gly in the nocturnal asthma group compared with the non-nocturnal asthma group (p < 0.01). However, there was no significant difference in the frequency of genotype Gly/Gly and allele Gly in the non-nocturnal asthma group, compared with the control group. There was no significance in the frequency of the genotypes and alleles of beta(2)-AR 27 loci among the three groups (p > 0.05). CONCLUSION: The Gly16 polymorphism of beta(2)-AR was overrepresented in nocturnal asthmatic patients, correlated with nocturnal asthma, and therefore appeared to be an important genetic factor in the expression of this asthmatic phenotype.     

Absence of autoantibodies to peptides shared by HLA-B27.5 and Klebsiella pneumoniae nitrogenase in serum samples from HLA-B27 positive patients with ankylosing spondylitis and Reiter''s syndrome.

Some microorganisms which are pathogenic in humans share amino acid sequences with human proteins (molecular mimicry). It has been suggested that molecular mimicry might be a reason for autoimmunity as a result of immunological cross reactivity. A homologous sequence of six amino acids has been found in both Klebsiella pneumoniae nitrogenase and the HLA-B27.5 molecule. In addition, (auto)antibodies to a synthetic peptide that contained the HLA-B27.5/klebsiella mimicking epitope have been detected in serum samples from HLA-B27 positive patients with ankylosing spondylitis and Reiter's syndrome. Confirmation of these data is important, because ankylosing spondylitis and Reiter's syndrome have so far been assumed to be 'seronegative' rheumatic diseases. It was, however, not possible to confirm the presence of autoantibodies against the mimicking peptide in serum samples from patients with ankylosing spondylitis and Reiter's syndrome. Serum samples from 81 patients with ankylosing spondylitis, 38 patients with Reiter's syndrome, and 81 healthy blood donors were tested against the 'mimicking peptide' in an enzyme linked immunosorbent assay (ELISA). Some of the serum samples from patients showed high but non-specific binding to the mimicking peptide. A highly significant correlation between binding to plastic coated with the mimicking peptide, to plastic coated with an irrelevant peptide, and even to non-coated plastic was observed. The nature of the serum component(s) in these patient serum samples (and some control serum samples) responsible for the high non-specific binding to plastic remains unclear. It was also shown that antibodies to the HLA-B27 peptide (containing the mimicking epitope) induced in rabbits do not cross react with the klebsiella peptide and vice versa.     

Systemic and mucosal antibodies to Klebsiella in patients with ankylosing spondylitis and Crohn''s disease.

Whole gut lavage fluid is a useful source of material for the study of intestinal immunity and inflammation in humans. Systemic and mucosal antibodies to Klebsiella pneumoniae were measured by enzyme linked immunosorbent assay (ELISA) in serum samples and whole gut lavage fluid from 14 patients with ankylosing spondylitis, 14 with Crohn's disease, and 16 immunologically normal controls. As the concentration of IgG in whole gut lavage fluid reflects disease activity in Crohn's disease, this approach was used to detect intestinal inflammation in patients with ankylosing spondylitis who also had disease activity and use of non-steroidal anti-inflammatory drugs (NSAIDs) recorded. Small intestinal permeability to cellobiose and mannitol was also studied. In serum samples, levels of IgA antibody to klebsiella were high in patients with Crohn's disease and in patients with active ankylosing spondylitis, and were significantly correlated with the erythrocyte sedimentation rate in patients with ankylosing spondylitis. Levels of IgG antibody to klebsiella were also high in patients with Crohn's disease. Studies of whole gut lavage fluid showed similar levels of IgA antibody to klebsiella in the three study groups, but levels of whole gut lavage fluid IgM and IgG antibodies to klebsiella were high in patients with Crohn's disease. Levels of IgG in whole gut lavage fluid were high in patients with Crohn's disease but in only one patient with ankylosing spondylitis, though the cellobiose/mannitol permeability ratio was abnormal in eight of 13 patients with ankylosing spondylitis. It is concluded that high levels of serum IgA antibody to klebsiella are not specific to ankylosing spondylitis, and that there is no evidence of an abnormal intestinal IgA antibody response to klebsiella in patients with ankylosing spondylitis.     

Cell-mediated immunity to collagen and collagen α chains in rheumatoid arthritis and other rheumatic diseases

Peripheral blood mononuclear cells from patients with rheumatoid arthritis, gout, ankylosing spondylitis and degenerative joint disease were cultured in the presence of native types I, II and III collagens and α chains from each of these types of collagen. The culture supernatant fluids were harvested and assayed for lymphocyte-derived chemotactic factor for monocytes. Reactions to one or more of the native collagens was found in 50 per cent (10 of 20) of the patients with rheumatoid arthritis, 20 per cent (two of 10) of the patients with gout and ankylosing spondylitis but in none of the 10 patients with degenerative joint disease or in normal subjects. Reaction to one or more α chains was found in 90 per cent (18 of 20) of the patients with rheumatoid arthritis, 60 per cent (six of 10) of the patients with gout, 50 per cent (five of 10) of the patients with ankylosing spondylitis, 30 per cent (three of 10) of the patients with degenerative joint disease and in 10 per cent of the normal subjects (one of 10). All the reactions were quantitatively stronger in patients with rheumatoid arthritis. These results indicate that patients with rheumatoid arthritis have cell-mediated immunity to homologous native and denatured collagens but that the reaction is not specific for rheumatoid arthritis. Some patients with gout, ankylosing spondylitis and degenerative joint disease also have low levels of immunity.     

A functional interleukin-10 mutation in Dutch patients with Crohn''s disease

BACKGROUND AND AIMS: Interleukin-10 is an anti-inflammatory and immunomodulatory cytokine. Interleukin-10 deficient mice are prone to develop chronic colitis. Administration of recombinant human interleukin-10 has been proposed to have a beneficial effect in a subgroup of patients with Crohn's disease. Recently, we found an interleukin-10 Gly15Arg mutation in a family with Crohn's disease which is associated with reduced interleukin-10 secretion by in vitro stimulated monocytes and lymphocytes. We hypothesised that this interleukin-10 mutation plays a role in maintaining the inflammatory process in Crohn's disease in some families. PATIENTS AND METHODS: We evaluated interleukin-10 Gly15Arg in 379 patients with Crohn's disease, and 75 unrelated healthy controls. Also, first degree family members of interleukin-10 Gly15Arg carriers were evaluated. Additionally, mutation carriers and their relatives were evaluated for CARD15 R702W, G908R, and 1007fs. RESULTS: Two patients with Crohn's disease were heterozygous for the interleukin-10 Gly15Arg mutation. No homozygotes were found. The Gly15Arg mutation was not observed in the controls. In first degree family members of the Crohn's disease-affected interleukin-10 Gly15Arg carriers, the mutation was found in Crohn's disease-affected as well as in their apparently healthy individuals. All family members carried one or two CARD15 mutation(s). CONCLUSION: The interleukin-10 Gly15Arg mutation is rare in patients with Crohn's disease, and is not associated with the disease in the Netherlands.