甘露糖结合凝集素基因点突变与风湿性心脏病易感性

管外科中心金振晓等,为探讨风湿性心脏病易感性与甘露糖结合凝集素基因第一外显子点突变的关系,采用PCR和限制性片段长度多态性分析对36例慢性风湿性心脏病病人和39例正常人甘露糖结合凝集素(MBL)等位基因分布情况进行分析。     

甘露糖结合凝集素基因多态性与中枢神经系统脱髓鞘疾病的关系研究简

目的 探讨甘露糖凝集素（mannosebinding lectin,MBL）基因多态性与中枢神经系统脱髓鞘疾病的相关性.方法 采用聚合酶链反应-限制性片段长度多态性分析（PCR-RFLP）法检测45例中枢神经系统脱髓鞘疾病和60例健康人的MBL基因多态性的分布.结果 45例中枢神经系统脱髓鞘疾病患者甘露糖凝集素基因Exon Ⅰ54位密码子点突变中,野生型（GGC/GGC）为34例,占75.56％;突变杂合子型（GGC/GAC）为10例,占22.22％;突变纯合子型（GAC/GAC）为1例,占2.22％.基因频率：GGC为86.67％,GAC为13.33％.对照组野生型（GGC/GGC）为30例,占50.00％;突变杂合子型（GGC/GAC）为28例,占46.67％;突变纯合子型（GAC/GAC）为2例,占3.33％.基因频率：GGC为73.33％,GAC为26.67％.中枢神经系统脱髓鞘疾病患者GGC/GGC基因型频率增高,而GGC/GAC基因型频率降低,差异均有统计学意义（P＜0.05）;中枢神经系统脱髓鞘疾病患者GGC等位基因频率高于健康对照组,而GAC等位基因频率低于健康对照组,差异有统计学意义（P＜0.05）.结论 MBL基因突变可能是中枢神经系统脱髓鞘疾病发病的危险因素.     

甘露糖结合凝集素等位基因变异与系统性红斑狼疮患者动脉血栓形成的风险

背景:心血管疾病是系统性红斑狼疮(SLE)患者的一项重要并发症。甘露糖结合凝集素等位基因与SLE及严重动脉粥样硬化有关。目的:确定甘露糖结合凝集素等位基因变异是否与SLE患者动脉血栓形成的风险增加相关。方法:在91例丹麦籍SLE患者中,通过PCR测定甘露糖结合凝集素等位基因的基因型。在前瞻性研究中评估SLE确诊后患者动静脉血栓形成情况。而动静脉血栓形成可以通过适当的诊断方法确定。结果:54例患者不具有甘露糖结合凝集素等位基因变异(即为正常基因型A/A),30例为变异杂合子(基因型A/O),7例为变异纯合子(基因型O/O)。在9.1年的中…     

热休克因子1基因剔除小鼠的遗传特征及基因型分析

为观察热休克因子 1基金剔除小鼠的遗传特征和基因型 ,摸索获得较多纯合子的繁殖技术 ,建立 3组繁殖组合方式 :1野生型♂×野生型♀组 ;2杂合子♂×杂合子♀组 ;3纯合子♂×杂合子♀组 ,并比较各组子代的基因型。结果表明 ,杂合子♂×杂合子♀组和纯合子♂×杂合子♀组 ,获得纯合子子代分别为 1 0 .8%和 2 2 .0 %。纯合子♂×杂合子♀组每胎产仔数 ( 4 .3只 /胎 )较野生型♂×野生型♀组 ( 6.5只 /胎 )及杂合子♂×杂合子♀组 ( 6.4只 /胎 )少。通过 HSF1剔除鼠的不同繁殖方法的比较及遗传特性观察 ,揭示了获得较多纯合子 HSF1剔除小鼠的繁殖技术。     

甘露糖结合凝集素基因多态性与肺结核的相关性研究

目的探讨甘露糖结合凝集素(MBL)基因多态性与肺结核的相关性。方法收集新乡医学院第一附属医院肺结核患者158例作为研究组,其生物学父母作为对照组。运用聚合酶链反应扩增及单核苷酸多态性的分子生物学技术,对MBL基因的rs69287125、rs137824326多态性分型,进行肺结核与甘露糖结合凝集素基因多态性的关联分析和单体型相对风险率分析。结果 rs69287125等位基因与肺结核相关联(P=0.011),其中等位基因G是保护因素(Z=-2.31),A为危险因素(Z=2.31);rs137824326等位基因与肺结核相关联(P=0.007),其中等位基因G是保护因素(Z=-2.54),A为危险因素(Z=2.54)。rs69287125-rs137824326单体型的A/A、G/G与肺结核有关联(P=0.021,0.015;Z=2.16,―1.85)。结论 MBL基因与肺结核相关联。     

血浆甘露糖凝集素和C反应蛋白与缺血性脑血管病的相关性

目的检测缺血性脑血管病(ICVD)患者甘露糖凝集素(MBL)基因外显子I区54位密码子(ExonI54)点突变情况,分析血浆MBL及C反应蛋白(CRP)含量与ICVD的相关性。方法ICVD组患者100例,健康对照组60例,用聚合酶链反应—扩增产物的限制性片段长度多态性分析(PCR-RFLP)法测试MBLExonI54点突变,用酶联免疫吸附试验(ELISA)测定MBL水平,用免疫比浊法测定CRP水平。结果ICVD组血浆MBL基因点突变频率分别为GGC84.0%、GAC16.0%,MBL平均含量为(3372±661)μg/L,CRP平均含量为(7.5±4.9)mg/L,健康对照组MBL基因点突变频率分别为GGC84.2%,GAC15.8%,MBL平均含量为(2065±196)μg/L,CRP平均含量为(2.2±1.3)mg/L。ICVD组与对照组基因点突变频率的比较差异无统计学意义(P>0.05)。ICVD组MBL水平、CRP含量均高于对照组(均P<0.01),CRP与MBL水平成正相关(γ=0.5,P<0.01),结论ICVD患者MBL点基因突变频率与健康者无显著差异。MBL和CRP可能参与ICVD的发生、发展。     

甘露聚糖结合凝集素基因多态性与毛细支气管炎的相关性研究

目的 探讨甘露聚糖结合凝集素(MBL)与毛细支气管炎是否具有相关性.方法 用ELISA法定量检测50例病例组与50例对照组MBL的血浆水平(ng/ml),并用聚合酶链反应、DNA序列测定法分析其MBL基因第1外显子第52、54、57位密码子的碱基序列.结果 (1)甘露聚糖结合凝集素GTC等位基因频率约为0.135,未见52和57位密码子的点突变;(2)比较病例组、对照组的GTC等位基因频率、血浆水平差异均无统计学意义.结论 未发现MBL的基因多态性与毛细支气管炎的易感性有相关性.     

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。     

宁夏回族人群甘露糖结合凝集素基因多态性研究

目的 调查宁夏回族人群中甘露糖结合凝集素(MBL)基因多态性,为进一步研究MBL基因多态性与疾病关联性提供线索。 方法 应用PCR-RFLP方法检测117例宁夏回族健康人群的MBL基因的多态性分布,并与宁夏汉族及其他民族健康人群中MBL基因多态性的分布进行比较。结果 在宁夏回族人群中,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,获得等位基因分布频率显示宁夏回族健康人群与宁夏汉族健康人群MBL基因多态性分布差异无显著性,而宁夏汉族与广东汉族相比较,差异有显著性。结论 本研究证实了同一人种内部MBL等位基因分布存在地理差异,也进一步支持中国汉族人群可分为南北两类的假设。     

CYPⅡE1基因多态性与药物性肝损害

目的 :研究细胞色素P4 5 0ⅡE1 (CYPⅡE1 )基因多态性与药物性肝损害发生发展的关系。方法 :从 4 3例药物性肝病患者及 5 0例正常人 (对照组 )外周血单个核细胞抽提基因组DNA ,采用PCR RFLP方法对CYPⅡE1基因分型。结果 :CYPⅡE1可分为A型 (c1纯合子 )、B型 (c1 /c2杂合子 )和C型 (c2纯合子 )。 5 0例正常人中c1基因频率 93% ,c2基因频率 7% ;4 3例药物性肝损害患者中c1基因频率 5 2 .3% ,c2基因频率 4 7.7% ,两组间差异显著 (P <0 .0 1 )。结论 :药物毒性反应的个体差异可能与CYPⅡE1等位基因的突变有关。     

风湿热患儿血清可溶性肿瘤坏死因子受体Ⅰ的变化

OBJECTIVE: To study the changes and the effects of soluble tumor necrosis factor receptor I (sT-NFR I) in patients with rheumatic fever, and to find out the laboratorial marker of clinic activity in patients with rheumatic fever. METHODS: The serum concentrations of sTNFR I in 16 patients with ARF, 16 patients with chronic inactive rheumatic heart disease (CRHD) and 15 healthy controls were measured by methods of ELISA and the relationship between sTNFR I and ESR or CRP were studied. RESULTS: The sTNFR I serum concentrations in patients with ARF (3.92 +/- 1.28) ng.ml-1 were significantly higher than those with CRHD (1.43 +/- 0.66) ng.ml-1 and healthy controls (1.06 +/- 0.42) ng.ml-1 (all P < 0.01). The serum concentrations of sTNFR I in patients with CRHD were almost the same with those in healthy controls (P > 0.05); Their positive correlation was significantly observed between sTNFR I and CRP(P < 0.001), and wasn't observed between sTNFR I and ESR. CONCLUSION: The Patients with ARF have high serum levels of sTNFR I. It may be used as the laboratorial marker in the clinic assessment of activity in patients with rheumatic fever.     

中国维吾尔族人群MBL基因 CGT52TGT、GGC54GAC和GGA57GAA点突变基因频率的研究

目的了解我国维吾尔族人群甘露聚糖结合凝集素(MBL)结构基因外显子1第52、54和57位密码点突变(CGT52TGT、GGC54GAC和GGA57GAA)的情况。方法收集新疆维吾尔族自治区维吾尔族一般人群血标本,提取白细胞基因组DNA,以PCR扩增目的基因片段并应用荧光探针杂交可视技术检测点突变。结果95例维吾尔族样本中,检出54位密码点突变纯合子2例和杂合子28例,未发现CGT52TGT和GGA57GAA点突变。结论维吾尔族人群MBL结构基因CGT52TGT、GGC54GAC和GGA57GAA点突变的频率分别为0、0.168和0。     

Homocysteine concentrations and molecular analysis in patients with congenital heart defects

BACKGROUND: Congenital heart defects are the result of incomplete heart development and, like many diseases, have been associated with high homocysteine concentration. METHODS: We evaluated homocysteine, folic acid and vitamin B(12) concentrations, and the mutations 677C>T and 1298A>C in MTHFR, 844ins68 in CBS and 2756A>G in MTR genes in 58 patients with congenital heart defects, 38 control subjects, and mothers of 49 patients and 26 controls. RESULTS: Control and patients presented normal range concentrations for homocysteine (7.66 +/- 3.16 microM and 6.95 +/- 3.12 microM, respectively), folic acid (8.31 +/- 3.00 ng/mL and 11.84 +/- 10.74 ng/mL) and vitamin B(12,) (613.56 +/- 307.57 pg/mL and 623.37 +/- 303.12 pg/mL), which did not differ among groups. For the mothers studied, homocysteine and vitamin B(12) concentrations also did not differ between groups. However, folic acid concentrations of mothers showed significant difference, the highest values being in the group of patients. No difference was found in allele frequencies among all groups studied. CONCLUSIONS: In the studied groups, high homocysteine seems not to be correlated with congenital heart defects, as well as folic acid and vitamin B(12). The mutations studied, in isolation, were not related to congenital heart defects, but high concentration of maternal homocysteine is associated with the presence of three or four mutated alleles.     

甘露糖结合凝集素在系统性红斑狼疮发病机制中的初步研究

目的通过检测系统性红斑狼疮(SLE)患者甘露糖结合凝集素(MBL)的血清水平及其基因变异情况,探讨MBL在SLE发病机制中的作用。方法以30名健康献血员为对照,以40例SLE患者为研究对象。通过固相ELISA方法检测MBL的血清水平。根据MBL2*LXPA(基因库X15422)单倍体序列设计了MBL外显子1区和启动子区的引物和荧光标记的寡核苷酸杂交探针,通过L ight Cyc ler实时PCR技术检测2组标本的基因变异情况。结果(1)SLE患者的MBL血清水平显著低于健康献血员(107.2μg/L对290.2μg/L,P<0.01)。(2)MBL基因外显子1区的突变以第54位密码子的突变为主,其突变率在SLE患者为37.1%(15/40),对照组为13.3%(5/30),两组比较χ2=3.914,P<0.05。(3)健康献血员和SLE患者在H/L位点均存在多态性,两组的L频率相当。(3)发生第54位密码子突变的SLE患者血清MBL水平显著下降,发生突变者与未发生突变者分别为141.7μg/L,49.8μg/L,两者比较P<0.01;两者的SLE活动指数(DAI)积分分别为12.87和7.44,P<0.01,且SLEDAI积分与MBL血清水平呈负相关(r=-0.48),启动子区H/L多态性与SLEDAI无关。结论发生MBL基因第54位密码子突变可能是SLE发病的易感因素;MBL的血清水平可能是SLE疾病活动性的潜在生物标记。     

风湿热患儿血清可溶性肿瘤坏死因子受体I的变化

目的 :了解肿瘤坏死因子系统在急性风湿热 (ARF)中的作用。方法 :用ELISA法对ARF、风湿性心脏病风湿活跃期 (活动期组 ) ,风湿性心脏病风湿静止期 (静止期组 )及健康人对照组血清可溶性肿瘤坏死因子受体I(sTN FRI)水平进行检测 ,并与血沉 (ESR) ,C反应蛋白 (CRP)进行相关分析。结果 :活动期组血清sTNFRI浓度为 (3 92±1 2 8)ng·ml- 1 明显高于静止期组 (1 43± 0 6 6 )ng·ml- 1 及健康人对照组 (1 0 6± 0 42 )ng·ml- 1 (均 P<0 .0 1 ) ;静止期组与健康人对照组比较 ,差异无显著性 (P >0 .0 5 )。活动期组血清sTNFRI改变与CRP呈正相关 (r =0 82 4 ,P <0 .0 0 1 )。结论 :ARF患儿血清sTNFRI水平可作为判断风湿活跃的一项实验室指标。     

家族性帕金森病Parkin基因外显子缺失突变

目的: 研究Parkin基因外显子缺失突变与中国家族性帕金森病(PD)发病的关系,探讨Parkin基因缺失突变在家族性PD发病机制中的作用及与临床特点、手术疗效之间的关系.方法: 家族性PD患者36例为研究对象,正常人20例作为对照,提取外周血白细胞基因组DNA,采用聚合酶链式反应(PCR)扩增目的DNA片段,琼脂糖凝胶电泳检测Parkin基因外显子4,6,7,10缺失情况,观察外显子的缺失分布及缺失率,并结合PD患者的手术疗效分析.结果: 家族性PD患者36例有10例外显子缺失突变,其中外显子4缺失8例,6和7缺失各1例,缺失突变率为27.8%,外显子10未发现异常,对照组中无缺失突变;有外显子缺失突变患者的手术疗效较好.结论: Parkin基因外显子4,6,7缺失突变是我国家族性PD的致病原因之一,PD患者的手术效果与Parkin基因外显子缺失突变与否有关.     

细胞毒性T淋巴细胞相关抗原-4基因多态性与Graves病复发的相关性研究

目的:探讨细胞毒性T淋巴细胞相关抗原(CTLA)-4基因多态性与Graves病(GD)复发的相关性.方法:选择102例GD复发患者(A组60例为停药3年内复发60例,B组为3年后复发42例)、40例GD初发患者和80名健康人群(正常对照组),应用聚合酶链反应-限制性内切酶片段长度多态技术进行CTLA-4基因外显子1第17密码子49位点A/G多态性检测.结果:CTLA-4基因第一外显子49位点基因型及等位基因的频率分布显示,GD初发组患者GG基因型频率及G等位基因频率均高于正常对照组(P<0.01).GD复发A组GG基因型频率均高于B组和初发组(P<0.01),B组GG基因型频率高于GD初发组(P<0.01).结论:CTLA-4基因外显子1第17密码子49位点A/G多态性与GD的发生存在相关性,该位点GG基因型可能是GD患者对抗甲状腺药物疗效差及早期复发的原因.     

Germline RET mutations in exons 10 and 11: an Iranian survey of 57 medullary thyroid carcinoma cases

The susceptibility gene for hereditary Medullary Thyroid Carcinoma (MTC) is the RET proto-oncogene. The aim of this study was to evaluate the prevalence of common germline RET mutations in exons 10 and 11 among Iranian MTC patients. Fifty-seven non-related MTC patients were examined in this study (Females: Males =1.2:1.0, Mean age = 40.0 +/- 11.5 years) and the existence of mutations was assessed through the PCR-RFLP technique. The only Multiple Endocrine Neoplasia type 2A (MEN2A) patient displayed a C634W mutation in exon 11. Among 53 apparently sporadic MTC patients, one patient showed a C620R mutation in exon 10 and two other patients displayed C624Y mutations in exon 11 of RET proto-oncogene. Neither the only Multiple Endocrine Neoplasia type 2B (MEN2B) patient nor two Familial MTC patients was found to carry germline mutations in exons 10 and 11. This study reports, for the first time, the prevalence of common RET mutations among Iranian, apparently sporadic MTC patients, underlining the critical importance of screening for RET mutations in such patients.     

HLA-DRB1 genes in 5 rheumatic disease multi-case families

OBJECTIVE: To detect HLA-DRB1 (DR1-10) alleles in 5 families with multi-case rheumatic diseases, and to study the possible influence of DRB1 genes in the pathogenesis of rheumatic diseases. METHODS: Sequence-Specific Primer PCR (PCR-SSP) method was used to examine HLA-DRB1 alleles. Totally 36 members of 5 families and 166 healthy people were involved in this study. The results were assessed by Chi-square test. RESULTS: The HLA-DRB1 allele frequency in the patients and their relatives was similar. No significant difference was found. But DR4 allele frequency in the patients (90.9%) and their relatives (68%) was much higher than that in normal controls (16.8%) and the difference was statistically significant (P < 0.0001). In family 4, two RA patients have different DRB1 alleles, while in family 5, two patients have the same DRB1 alleles, one developed SLE and the other developed RA. CONCLUSIONS: DR4 is closely related to rheumatoid arthritis. The nelatives of RA patients may be at greater risk to develop RA than individuals without family history. Some patients had the same DRB1 allele but developed different rheumatic diseases. This suggested that there might be some common pathways in genetic predisposing of rheumatic diseases. On the other hand, only a few patients with the same DRB1 allele developed rheumatic diseases during their life, so other factors besides DRB1 gene might also be involved in the pathogenesis of rheumatic diseases.     

HBsAg和HBsAb双阳性慢性乙肝患者血清中HBV基因型与S区突变的关系

目的 探讨HBsAg和HBsAb同时阳性（HBsAg＋/HBsAb＋）慢性乙肝患者血清中HBV基因型与S区氨基酸序列突变的关系.方法 将43例HBsAg＋/HBsAb＋慢性乙肝患者作为实验组,并选取35例HBsAg＋/HBsAb-慢性乙肝患者作为对照组,对血清中HBV DNA S区基因序列进行PCR扩增和测序,根据测序结果对患者的基因型进行分型并对S区氨基酸序列突变进行比对分析.结果 实验组中B基因型8例、C基因型35例,对照组中B基因型9例、C基因型26例.实验组C基因型患者的年龄[（50.2±16.3）岁]明显大于B基因型[（34.4±13.4）岁]（P=0.015）;实验组B基因型和C基因型的S区氨基酸突变率分别为0.77%和1.64%（P=0.005）;而对照组的B、C两种基因型的突变率分别为0.59%和0.49%（P=0.597）.同样为C基因型的两组患者,实验组S区氨基酸突变明显多于对照组（P=0.000）,而且实验组S区氨基酸突变大多分布在主要亲水结构区域（MHR）α决定簇的第一个环状结构区内;而B基因型只有少数患者有α决定簇内的突变.结论 HBsAg＋/HbsAb＋的乙肝患者C基因型较B基因型更易发生突变;且C基因型患者的氨基酸突变大多发生于S区的α决定簇内,可能正是由于此种突变使HbsAg的抗原性发生改变,最终导致了HbsAg和HbsAb同时阳性.