贵州三都水族居民葡萄糖-6-磷酸脱氢酶缺乏症基因突变研究

为了了解贵州省少数民族居民葡萄糖-6-磷酸脱氢酶(glucose-6-phosphate dehydrogenase，G6PD)缺乏症的发病率、基因突变类型特点及分布特征，进一步从分子水平揭示G6PD基因突变的异质性，对贵州省三都水族自治县1090名当地水族居民采用噻唑蓝定性法、G6PD／6PGD活性比值法进行G6PD缺乏症的筛查，再经错配引物介导的聚合酶链反应/限制性酶切分析法检测中国人中最常见的3种G6PD基因突变型：1376G→T、1388G→A、95A→G。结果表明：在受检的1090人中，共检出G6PD缺乏症98例，检出率8．99％，在G6PD缺乏症中检出最常见的3种G6PD基因突变型：1376G→T24例；1388G→A12例；95A→G9例；并在国内首次检出1376G→T、95A→G复合型突变1例。1376G→T突变频率为0．245；1388G→A突变频率为0．122；95A→G突变频率为0．092。结论：1376G→T、1388G→A、95A→G为贵州省三都水族居民的常见G6PD突变型，这个结果提示贵州三都水族与中国其它少数民族在起源上可能有共同的渊源。     

急性白血病患儿葡萄糖-6-磷酸脱氢酶活性的探讨

白血病是一组造血干细胞恶性克隆性疾病,可存在多种红细胞酶和同工酶的异常.我们检测了82例急性白血病患儿红细胞葡萄糖-6-磷酸脱氢酶（G6PD）活性,并探讨其临床意义,现报道如下.     

葡萄糖-6-磷酸脱氢酶两种测定方法的比较

目的对两种检测全血中红细胞葡萄糖-6-磷酸脱氢酶(G6PD)活性的方法进行比较分析,观察其优、缺点和相关性。方法每天分别用两种方法对25例患者标本进行检测,连续检测3d,共75例,并对检测结果进行比较分析,观察其相关性。结果手工定量比值法有2例判为轻度缺乏的标本,仪器紫外速率法的结果为186U/L和251 U/L;手工定量比值法有4例重度缺乏的标本,仪器紫外速率法的结果均小于150U/L;其余手工定量比值法正常(比值大于1.0)的标本,仪器紫外速率法的结果均大于500U/L。结论两种方法结果一致,相关性较好,但各有优缺点。     

浙江省葡萄糖-6-磷酸脱氢酶缺乏症筛查现况调查

目的探讨新生儿葡萄糖-6-磷酸脱氢酶(G6PD)活性的流行病学分布特征、G6PD缺乏症患病率及cut-off值水平。方法选取2015年3月至2017年9月,浙江省10个地市共计约144万例新生儿G6PD筛查数据。用荧光分析法测定滤纸血片中G6PD活性,初筛阳性者召回并用G6PD/6PGD直接比值法确证。用非参数检验和卡方检验等方法进行统计分析。结果不同性别、出生孕周、出生体重、采血时年龄和采血季节等因素下,G6PD活性分布差异有统计学意义(P0.01)。浙江省G6PD缺乏症男性患病率高于女性,丽水市患病率最高(0.38%),舟山市患病率最低(0.11%),在浙江省内基本呈现南高北低的趋势。当G6PD活性cut-off值在2.60～2.80 U/g Hb范围时,男、女性新生儿G6PD缺乏症筛查的敏感性均为100%,且约登指数也最高(约为0.99)。结论 G6PD活性与人群和时间等因素有关,G6PD缺乏症患病率受性别、地区影响较大,建议浙江省G6PD筛查cut-off值定为男性2.60 U/g Hb,女性2.80 U/g Hb。     

应用荧光法筛查葡萄糖6-磷酸脱氢酶缺乏

目的建立适于葡萄糖6-磷酸脱氢酶缺乏筛查的检测方法。方法用荧光法对新生儿筛查滤纸干血片标本进行检测,对阳性者召回抽静脉血以G6PD/6PGD比值法进行确诊。结果用荧光法测定7780份新生儿筛查滤纸血片标本的G6PD活性,阳性358例,阳性率为4.6%(358/7780),对召回的358例阳性以比值法进行确诊试验,阳性315例,阳性率为4.0%,两者符合率为87.9%(315/358)。结论荧光法是一种半定量的方法,具有较高的特异性和准确性,且操作简便、检测快速、成本低廉,适合用滤纸干血片标本进行大规模的新生儿筛查。     

新生儿黄疸葡萄糖-6-磷酸脱氢酶活性检测及临床意义

目的通过新生儿葡萄糖-6-磷酸脱氢酶(G-6-PD)活性检测,了解耒阳地区新生儿黄疸患儿G-6-PD缺乏情况,为新生儿黄疸的临床诊断及治疗提供科学依据。方法对235例新生儿黄疸患者进行血浆G-6-PD活性测定。结果 235例新生儿中G-6-PD缺乏者4例,阳性率为1.7%。结论 G-6-PD缺乏为新生儿黄疸的重要原因,对G-6-PD缺乏的新生儿进行早期干预,能有效减轻G-6-PD缺乏,从而减轻新生儿溶血的程度,避免发生核黄疸。     

我国葡萄糖-6-磷酸脱氢酶缺乏症的分布特征和基因突变

葡萄糖-6-磷酸脱氢酶(glucose-6-phosphate dehydrogenas,G6PD)缺乏症是最常见的遗传性酶缺陷疾病,可导致急性溶血性贫血、新生儿高胆红素血症和慢性非球形红细胞溶血性贫血等疾病,其发生率和基因突变类型有明显的地域和种族特异性,本文主要综述了中国不同民族和地区G6PD缺乏症的分布特点和基因突变.     

葡萄糖-6-磷酸脱氢酶缺乏症的分子机制及诊断方法研究进展

葡萄糖-6-磷酸脱氢酶(G6PD)是催化6-磷酸葡萄糖脱氢,以维持抗氧化物质谷胱甘肽(GSH)的还原性,清除细胞内过氧化物的毒性,保护血红蛋白及细胞膜巯基蛋白,而维持红细胞结构和功能的稳定。G6PD缺乏症是全球最常见的X连锁不完全显性遗传性酶缺陷综合征,俗称蚕豆病,全球约4亿人受累,患者在食用蚕豆、服用氧化性药物和感染等情况下,血红蛋白氧化变性,红细胞破坏而导致溶血[1]。中国南方地区为此病高发区。现对G6PD缺乏症的临床症状、人群分布、发病机     

酶耦联法测定葡萄糖-6-磷酸脱氢酶的研究

目的建立葡萄糖-6-磷酸脱氢酶的酶耦联测定法。方法用葡萄糖-6-磷酸作为底物启动酶促反应,340nm连续监测NADPH生成速率。结果此酶的反应的最适pH9．0,方法线性可达1000U／L,批内变异系数（CV）为3％～8％,回收率为98．9％～101.5％。结论葡萄糖-6-磷酸脱氢酶的酶耦联测定法与既往所述的单机法比较具有良好的相关性,相关系数（r）=0．99,y=0．989X＋0．03,     

武汉地区葡萄糖-6-磷酸脱氢酶缺乏症患者基因突变分析

目的:探讨武汉地区葡萄糖-6-磷酸脱氢酶(G6PD)缺乏症患者的基因突变特征。方法:对湖北省妇幼保健院1 321例筛查阳性的新生儿及门诊患者进行G6PD基因检测,首选多色探针熔解曲线法(MMCA)检测中国人群常见的12种G6PD基因突变,对于MMCA法未检出者,分析其酶活性和临床信息,必要时进行测序检测。结果:在1 321例受检者中共检出768例突变,检出率58.1%。共检出18种G6PD基因变异类型,包括c.1388G>A、c.1376G>T、c.95G>A、c.1024C>T、c.871G>A、c.392G>T、c.487 G>A、c.1360C>T、c.1004C>A、c.517T>C、c.592C>T、c.94C>G、c.152C>T、c.320A>G、c.1028A>G、c.1316G>A、c.1327G>C和c.1376G>C,其中男性半合子683例,女性纯合子3例,女性杂合子80例,女性复杂杂合2例。结论:本研究共检出18种G6PD基因变异类型,并首次在中国人群中报...     

O1 and non-O1 Vibrio cholerae bacteremia produced by hemolytic strains

Vibrio cholerae are Gram-negative bacteria capable of producing serious infections. They are differentiated into O1 and non-O1 serogroups, depending on their ability to agglutinate with specific antiserum. In contrast to non-O1 V. cholerae, which are more prone to invading the bloodstream, V. cholerae O1 is rarely the cause of bacteremia. We describe 2 cases of O and non-O1 V. cholerae bacteremia in patients with hepatitis C virus cirrhosis. We postulate that the hemolytic properties of the isolates contributed to their virulence in immunocompromised hosts.     

Vibrio cholerae O1 serotype Ogawa in a neonate

In India, cholera is endemic and affects usually the 3 to 5-year-old age group. There have been occasional reports in the neonatal period with Vibrio cholerae O139 Bengal. We report here a case of Vibrio cholerae O1 diarrhea in a 2-day-old, breastfed male, who had been delivered in the hospital and developed severe dehydration.     

间日疟原虫乳酸脱氢酶GST融合表达载体的构建及表达

目的构建间日疟原虫乳酸脱氢酶（PvLDH）融合表达载体,并表达PvLDH的GST融合蛋白。方法将PvLDH基因片段克隆到表达载体pGEX-4T-1中,构建PvLDH/pGEX-4T-1融合表达载体,IPTG诱导表达目的基因,SDS-PAGE电泳分析表达产物,Western blot检测其抗原性。结果成功构建了PvLDH/pGEX-4T-1融合表达系统,在大肠杆菌BL21中以包涵体形式高效表达,表达产物能与恶性疟原虫和间日疟原虫感染患者血清反应,而不与正常人血清反应。结论间日疟原虫LDH蛋白在大肠杆菌中获得高效表达,表达产物具有良好的抗原性。     

CDCP1胞外段基因的克隆及在大肠杆菌中表达

目的：构建携带 CDCP1胞外段基因的原核表达载体。方法以 A549的 mRNA 为模板,应用所设计的引物通过 RT-PCR 法扩增 CDCP1胞外段基因;将 PCR 产物与 pGEX-KG 载体连接,获得重组质粒 pGEX-KG/CDCP1;经双酶切、PCR 及 DNA 序列测定进行鉴定;IPTG 诱导表达。结果①RT-PCR 扩增得到约270 bp 大小的 CDCP1胞外段基因目的片段;②目的片段正确插入到 pGEX-KG 中;③经 IPTG 诱导表达,可见在相对分子质量约35kD 处出现明显的诱导蛋白条带,与预期一致。结论成功构建了携带 CDCP1胞外段基因的原核表达载体,在大肠杆菌中获得大量表达。     

Experimental non-O group 1 Vibrio cholerae gastroenteritis in humans.

In this study, 27 volunteers received one of three non-O group 1 Vibrio cholerae strains in doses as high as 10(9) CFU. Only one strain (strain C) caused diarrhea: this strain was able to colonize the gastrointestinal tract, and produced a heat-stable enterotoxin (NAG-ST). Diarrhea was not seen with a strain (strain A) that colonized the intestine but did not produce NAG-ST, nor with a strain (strain B) that produced NAG-ST but did not colonize. Persons receiving strain C had diarrhea and abdominal cramps. Diarrheal stool volumes ranged from 154 to 5,397 ml; stool samples from the patient having 5,397 ml of diarrhea were tested and found to contain NAG-ST. The median incubation period for illness was 10 h. There was a suggestion that occurrence of diarrhea was dependent on inoculum size. Immune responses to homologous outer membrane proteins, lipopolysaccharide, and whole-cell lysates were demonstrable with all three strains. Our data demonstrate that V. cholerae of O groups other than 1 are able to cause severe diarrheal disease. However, not all strains are pathogenic for humans: virulence of strain C may be dependent on its ability both to colonize the intestine and to produce a toxin such as NAG-ST.     

一起输入性霍乱弧菌的实验室检测

目的分离培养患者粪便标本中的霍乱弧菌并检测其毒力基因及药物敏感性。 方法常规方法分离菌株，根据其生化及血清学反应结果鉴定菌株，聚合酶链反应 (PCR) 检测毒力基因IctxA、ace、tcpA、zot，/IK-B法测定药物敏感性。 结果 从患者粪便中分离到埃尔托生物型6f霍乱弧菌1株，带有4种毒力基因，对多种药物敏感，不产-内酰胺酶。 结论结果提示该株霍乱弧菌为流行株，对多种常用药物敏感，PCR可较快完成对霍乱弧菌毒力强弱的检测。     

GM1-dot-EIA for the detection of toxin-producing Vibrio cholerae strains

A new variant of enzyme immunoassay (EIA) has been developed on the basis of GM1 gangliosides to detect the toxin-producing Vibrio cholerae strains--GM1-dot-EIA. Experiments were run using a nitrocellulose membrane to bind GM1 gangliosides and polyclonal antitoxic serum to detect cholerogen. GM1-dot-EIA testing identified cholera toxin in 11 of 13 supernatants of V. cholerae eltor ctx(+) strains isolated from man and in 3 of 7 supernatants of V. cholerae eltor ctx(+) strains isolated from water. These data agree with those obtained in CM1-EIA. There was no reaction with the supernatants of other microorganisms. The sensitivity of the technique was 10 ng/ml. Thus, the simple and specific GM1-dot-EIA may be recommended to detect toxin-producing V cholerae strains isolated from man and water.     

Fatalities associated with Vibrio parahaemolyticus and Vibrio cholerae non-O1 infections in Florida (1981 to 1988)

Vibrio infections constitute a continuing source of morbidity and mortality in Florida. Seven fatal infections caused by Vibrio parahaemolyticus or V cholerae non-O1 were reported in Florida between 1981 and 1988. Review of those seven medical records and Vibrio case investigation forms showed that although all patients died of sepsis, gastrointestinal signs and symptoms characterized the early illness in four patients, whereas the other three initially had painful swelling and/or lesions of the lower extremities. All patients had preexisting chronic diseases. Five patients (71%) had eaten seafood during the week before oneset of illness, including four (57%) who had eaten raw oysters. To reduce the risk of acquiring Vibrio infections, raw or undercooked seafood should be eliminated from the diet, particularly by persons with underlying chronic diseases.     

霍乱弧菌的分子分型方法

在霍乱流行和暴发调查中,追溯传染源和调查传播途径往往需要对霍乱弧菌进行分型分析.对霍乱弧菌进行血清分型和噬菌体一生物分型,是得到菌株基本信息不可缺少的手段.如果想要揭示菌株在分子水平上的变异和进化规律,则需要进行分子分型分析.本研究对霍乱弧菌的各种分子分型方法进行逐一介绍并加以综合比较.