上海地区汉族人HLA-DQB1启动子多态性以及QBP-DQB1连锁分析

为分析上海地区汉族人HLA DQB1启动子 (QBP )多态性 ,以及QBP与DQB1的连锁不平衡 ,我们选用 2 4个序列特异性寡核苷酸探针 (SSO )检测QBP多态性。在目前已知的 12种QBP等位基因中 ,我们仅检测到 10种 (未发现新的QBP类型 ) ,并且发现QBP与DQB1的某些等位基因之间存在一定的连锁格局。我们的结果与高加索人群研究结果比较发现 ,无论是QBP基因频率还是QBP DQB1单元型频率 ,在两种不同人群中都存在明显差异。结果提示 ,由于不同的QBP等位基因功能不同 ,因此QBP DQB1之间不同的单倍型组合可能是HLA与疾病关联的另一个因素。     

上海地区汉族人HLA—DQB1启动子多态性以及QBP—DQB1连锁分析

为分析上海地区汉族人HLA－DQB1自动子（QBP）多态性,以及QBP与DQB1的连锁不平衡,我们选用24个序列特异性寡核苷酸探针（SS0）检测QBP多态性。在目前已知的12种QBP等位基因中,我们仅检测到10种（未发现新的QBP类型）,并且发现QBP与DQB1的某些等位基因之间存在一定的连锁格局。我们的结果与高加索人群研究结果比较发现,无论是QBP基因频率还是QBP－DQB1单元型频率,在两种不同人群中都存在明显差异。结果提示,由于不同的QBP等位基因功能不同,因此QBP－DQB1之间不同的单倍型组合可能是HLA与疾病关联的另一个因素。     

不明原因习惯性流产与肿瘤坏死因子水平及肿瘤坏死因子α-308基因多态性的相关性初步研究

目的 研究肿瘤坏死因子（TNF）水平及肿瘤坏死因子α-308基因多态性与不明原因习惯性流产的关系。方法 用放免法测定50例不明原因习惯性流产患者及50例健康对照者血清TNF水平，用聚合酶链反应扩增TNF-α-308基因片段，NcoI内切酶进行限制性酶切反应。结果 50例妊娠高血压综合征患者及50例正常对照TNF-α-308基因型及等位基因频率均无显著性差异，而血浆中TNF水平，不明原因习惯性流产组明显高于对照组（P〈0．05）。结论 炎性反应在不明原因习惯性流产病程中有重要作用，TNFα-308基因多态性与不明原因习惯性流产无关联。     

不明原因习惯性流产与肿瘤坏死因子水平及肿瘤坏死因子α-308基因多态性的相关性初步研究

目的研究肿瘤坏死因子(TNF)水平及肿瘤坏死因子α-308 基因多态性与不明原因习惯性流产的关系.方法用放免法测定50例不明原因习惯性流产患者及50例健康对照者血清TNF水平, 用聚合酶链反应扩增TNF-α-308 基因片段,NcoI内切酶进行限制性酶切反应.结果 50例妊娠高血压综合征患者及50例正常对照TNF-α-308基因型及等位基因频率均无显著性差异, 而血浆中TNF水平,不明原因习惯性流产组明显高于对照组(p<0.05).结论炎性反应在不明原因习惯性流产病程中有重要作用,TNFα-308基因多态性与不明原因习惯性流产无关联.     

HLA-DRB1、DQB1基因与汉族人群寻常型天疱疮的相关性研究

目的　探讨 HL A- DRB1、DQB1位点基因在汉族人群寻常型天疱疮易感性中的作用。方法用序列特异性引物 -聚合酶链反应方法 ,对 6 1例寻常型天疱疮 (pemphigus vulgaris,PV)患者和 5 7名正常对照进行了 HL A- DRB1、DQB1等位基因的分型 ,并分析了 DRB1、DQB1基因在两组中的分布。结果　与正常对照组比较 ,PV组 DR4、DRB1* 14 (* 14 0 1、* 14 0 4、* 14 0 5 )基因频率明显增高 (Pc分别 <0 .0 5及P<0 .0 1) ,差异有显著性 ;PV组 DQB1* 0 5 0 3、DQB1* 0 30 2基因频率明显增高 (Pc均 <0 .0 5 ) ,差异有显著性。对 DR4阳性样本的组内基因亚型分型结果发现 ,PV组中 DRB1* 0 4 0 3、DRB1* 0 4 0 6频率显著增高(Pc<0 .0 5 ) ,差异有显著性。 PV患者组单倍型 HL A- DRB1* 0 4 ,DQB1* 0 30 2和 HL A- DRB1* 14 ,DQB1* 0 5 0 3频率明显增高 (P<0 .0 5 )。结论　HL A- DRB1* 0 4 ,DQB1* 0 30 2和 HL A- DRB1* 14 ,DQB1* 0 5 0 3可能是汉族人 PV推测的易感单倍型。     

HLA—E与原因不明习惯性流产的关联研究

目的探索HLA-E与原因不明习惯性流产的发生是否存在关联.方法采用地高辛标记的寡核苷酸探针杂交技术,对上海地区53例原因不明习惯性流产病人和156例正常对照进行了HLA-E等位基因检测.结果在病人和对照组中HLA-E*0101均是最常见的等位基因,其基因频率分别为50%和43.09%,其次是E*01032分别为29.25%和33.12%.E*01031则分别占20.75%和23.79%,HLA-E*0102和E*0104在两组标本中均未检出.检出的3个等位基因在两组之间比较均无显著性差异(P＞0.05).结论HLA-E等位基因多态性与原因不明习惯性流产的发生可能没有直接关联.     

PAI-1,FV,FⅡ和MTHFR基因多态性对习惯性流产的影响

目的探讨血栓形成因子基因多态性与习惯性流产的关系。方法通过快速循环PCR和荧光共振能量转移技术分析纤溶酶原激活物抑制剂(Plasminogen activator inhibitor-1,PAI-1)5G/4G、凝血因子V(Factor V,FV)G1691A、凝血因子Ⅱ(FactorⅡ,FⅡ)G20210A和亚甲基四氢叶酸还原酶(Methylenetetrahydrofolate reductase,MTHFR)C677T的基因多态性在130例习惯性流产妇女和130例健康妇女之间分别频率的差异。结果在习惯性流产妇女中,FV和FII G/A杂合基因型频率(FV:11.5%,FⅡ:15.4%)显著高于对照组(FV:3.8%,FⅡ:2.3%)(P0.01);等位基因A的突变频率(FV:5.8%,FⅡ:7.7%)显著高于对照2组(FV:1.9%,FⅡ:1.2%)(P0.01)。PAI-1 5G/4G杂合子频率(40.8%)显著高于对照组(23.8%)(P0.05),且病例组纯合子4G/4G、5G/5G和等位基因频率与对照组之间没有显著性差异。病例组MTHFR C677T的基因型和等位基因频率与对照组之间没有显著性差异。结论 FV G1691A和FII G20210A G/A杂合基因型是习惯性流产发生的高危险因素,PAI-1 5G/4G杂合基因型是习惯性流产发生的低危险因素,而MTHFR C677T基因型与习惯性流产发生没有直接关系。     

白细胞介素Ⅰ受体拮抗剂基因多态性与习惯性流产的关系

目的 探讨白细胞介素Ⅰ受体拮抗剂基因(interleukin-1 receptor antagonist gene,IL1RA)多态性与原因不明习惯性流产的相关性.方法 应用聚合酶链反应技术,检测了97例习惯性流产患者(患者组)和99名正常孕妇(对照组)的白介素Ⅰ受体拮抗剂基因进行扩增,并用琼脂糖凝胶电泳观察结果.*1为%,IL1RA 3/3 91.8%,等位基因分布:IL1RA结果习惯性流产患者基因型分布:IL1RA 1/3 8.2*3为95.9%;正常孕妇基因型分布:IL1RA 1/3为8.1%,IL1RA 3/3为90.9%,IL1RA 3/4%,IL1RA4.1*1为4.0%,IL1RA*3为95.5%,IL-1RA*4为0.5%,两组间差异无统%,等位基因分布:IL1RA为1.0计学意义(P>0.05).结论 原因不明习惯性流产患者IL1RA基因多态性的分布与正常孕妇无明显相关性.     

白细胞介素Ⅰ受体拮抗剂基因多态性与习惯性流产的关系

目的 探讨白细胞介素Ⅰ受体拮抗剂基因(interleukin-1 receptor antagonist gene,IL1RA)多态性与原因不明习惯性流产的相关性.方法 应用聚合酶链反应技术,检测了97例习惯性流产患者(患者组)和99名正常孕妇(对照组)的白介素Ⅰ受体拮抗剂基因进行扩增,并用琼脂糖凝胶电泳观察结果.*1为%,IL1RA 3/3 91.8%,等位基因分布:IL1RA结果习惯性流产患者基因型分布:IL1RA 1/3 8.2*3为95.9%;正常孕妇基因型分布:IL1RA 1/3为8.1%,IL1RA 3/3为90.9%,IL1RA 3/4%,IL1RA4.1*1为4.0%,IL1RA*3为95.5%,IL-1RA*4为0.5%,两组间差异无统%,等位基因分布:IL1RA为1.0计学意义(P>0.05).结论 原因不明习惯性流产患者IL1RA基因多态性的分布与正常孕妇无明显相关性.     

白细胞介素Ⅰ受体拮抗剂基因多态性与习惯性流产的关系

目的 探讨白细胞介素Ⅰ受体拮抗剂基因(interleukin-1 receptor antagonist gene,IL1RA)多态性与原因不明习惯性流产的相关性.方法 应用聚合酶链反应技术,检测了97例习惯性流产患者(患者组)和99名正常孕妇(对照组)的白介素Ⅰ受体拮抗剂基因进行扩增,并用琼脂糖凝胶电泳观察结果.*1为%,IL1RA 3/3 91.8%,等位基因分布:IL1RA结果习惯性流产患者基因型分布:IL1RA 1/3 8.2*3为95.9%;正常孕妇基因型分布:IL1RA 1/3为8.1%,IL1RA 3/3为90.9%,IL1RA 3/4%,IL1RA4.1*1为4.0%,IL1RA*3为95.5%,IL-1RA*4为0.5%,两组间差异无统%,等位基因分布:IL1RA为1.0计学意义(P>0.05).结论 原因不明习惯性流产患者IL1RA基因多态性的分布与正常孕妇无明显相关性.     

HLA-DRB1等位基因多态性与原因不明习惯性流产相关性研究

目的探讨HLA-DRB1等位基因多态性与原因不明习惯性流产（UHA）相关性。方法采用PCR-SSP方法对36对UHA夫妇（UHA组）进行HLA-DRB1等位基因分型,并与34对正常夫妇（对照组）比较,将结果进行统计学处理。结果HLA-DRB1＊11等位基因在UHA组中的频率分布较对照组明显增高P〈0.05,HLA-DRB1＊04等位基因在UHA组中的频率分布较对照组明显降低P〈0.05;UHA组中妻子HLA-DRB1＊08等位基因频率分布较对照组妻子明显增高P〈0.05,HLA-DRB1＊07等位基因频率分布较对照组妻子明显降低P〈0.05。结论本地区汉族人群HLA-DRB1等位基因与UHA相关,HLA-DRB1＊11等位基因可能是UHA夫妇的易感基因;HLA-DRB1＊08等位基因可能是UHA妻子的易感基因。     

Possible susceptibility of the HLA-DPB1*0402 and HLA-DPB1*04 alleles to unexplained recurrent abortion: analysis by means of polymerase chain reaction-restricted fragment length polymorphism method

PROBLEM: To clarify whether HLA-DP antigens are associated with patient population of unexplained recurrent abortion. METHOD OF STUDY: The frequency of HLA-DPB1 alleles in patients with unexplained recurrent abortion, and the compatibility of HLA-DPB1 alleles between patient couples, were studied using a polymerase chain reaction (PCR)-restricted fragment length polymorphism (RFLP) method. Thirty patients who had a history of unexplained primary recurrent abortion, and their husbands, were typed for HLA-DPB1 genotype. Two hundred and ninety-nine base pair fragments from the second exon of HLA-DPB1 genes were selectively amplified using the PCR-primers. After amplification, the DNAs were digested with restriction endonucleases, and subjected to electrophoresis in a 12% polyacrilamide gel to determine HLA-DPB1 genotype. RESULTS: The frequency of HLA-DPB1*0402 and DPB1*04 alleles in the patient group (n = 30) was significantly increased, as compared to that in the normal fertile women (n = 30). The frequency of HLA-DPB1*04 allele in the patient group was significantly increased, as compared to that in the general population (n = 112). No significant compatibility of HLA-DPB1 alleles could be observed between patient couples and normal fertile couples. CONCLUSION: These findings suggest a possible new class II association with patient population of unexplained recurrent abortion.     

The promotor QAP and QBP alleles in Czech population and rheumatoid arthritis multicase families

HLA promotor alleles of DQA1 and DQB1 genes were analyzed in a group of 65 Czech healthy individuals and 58 members of 14 RA multicase families by PCR and oligonucleotide hybridization. QAP4.1 and QBP3.1 were the most frequent alleles (gf=0.315, GF=0.254) due to the linkage disequilibrium with DQA1×0501 and DQB1×0301. The second most frequent allele QAP1.2 (gf=0.162) was found to be common for DR2 alleles. The analysis of RA multicase families showed promotor alleles and haplotypes similar to those reported in the healthy population. QBP3.21 was in a commplete positive association with DQB1×0302-DRB1×04, except DQB1×0302-DRB1×0408, where was detected QBP3.1, associated also with DQB1×0301-DRB1×0401. Promotor polymorphism may reflect a level of expression and function of HLA structural alleles.     

复发性流产和不明原因不孕患者子宫内膜白血病抑制因子表达研究

目的探讨白血病抑制因子(leukaemia inhibitory factor,LIF)在复发性流产和不明原因不孕患者黄体中期子宫内膜的表达情况.方法采用实时荧光定量PCR技术,对31例不明原因不孕和30例复发性流产患者黄体中期子宫内膜LIF表达量的测定,对照组34例为继发性不孕患者(输卵管阻塞的)黄体中期子宫内膜.结果不明原因不孕组妇女子宫内膜中LIF表达量(M)为(169.00ng/l),与对照组(M) 为(240.00ng/l)相比,差异有显著性(P<0.01);习惯性流产组LIF表达量(M) 为(90.25ng/l),与对照组相比,差异有显著性(P<0.01),复发性流产组与不明原因不孕组相比差异无显著性(P>0.05).结论 LIF基因可能在启动胚泡着床和维持妊娠方面有重要作用, 黄体中期子宫内膜LIF基因表达量的减少可能是导致不明原因不孕和复发性流产的原因.     

Polymorphism in the upstream regulatory region of the DQB1 gene (QBP) and four point (DRB1, QBP, DQA1, DQB1) haplotypes in the Dai minority population in China.

Using polymerase chain reaction and Dig-ddUTP labeled oligonucleotides we have investigated the DNA polymorphism for the DQB1 promoter region (QBP) and we have deduced four point haplotypes in 65 unrelated healthy individuals of the Dai minority population. A total of 8 QBP alleles were detected. The most frequent allele is QBP 5.11 with 87.7% allele frequency followed by QBP 3.21, 3.1, 5.12 with 33.8%, 23.1% and 15.4% allele frequency respectively. Four QBP alleles, 3.22, 3.32, 4.1 and 6.12 were absent in the Dai minority. The linkage disequilibrium of the QBP allele with certain DQB1 alleles was very strong. Complete positive association was found for QBP 2.1-DQB1^*0201, QBP3.1-DQB1^*0301, QBP6.11-DQB1^*0601. A total of 32 different four point haplotypes were deduced. Among them the most common haplotypes were DRB1^*1602, DQA1^*0102, QBP5.11; DQB1^*0502, DRB1^*1602-QBP5.11, DQA1^*0102, DQB1^*0502 (N = 19); DRB1^*09 DQA1^*03, QBP3.21, DQB1^*03032 (N = 5); DRB1^*1401, DQA1^*01, QBP5.11, DQB1^*0502 (N = 12) and DRB1^*1202, DQA1^*0601, QBP3.1, DQB1^*0301 (N = 10). We conclude from these data that a) there is a reduced class II polymorphism in the Dai minority population an b) the relationship between QBP and DQB1 alleles is not different from that observed in other populations.     

Association of the A/G polymorphism at position 49 in exon 1 of CTLA-4 with the susceptibility to unexplained recurrent spontaneous abortion in the Chinese population

PROBLEM: To investigate whether the A/G polymorphism at position 49 in exon 1 of cytotoxic T lymphocyte antigen-4 (CTLA-4) gene, which delivers a negative signal to T-cell activation, confers the susceptibility to unexplained recurrent spontaneous abortion in the Chinese population. METHOD OF STUDY: A total of 168 patients with unexplained recurrent spontaneous abortion (RSA), who were treated in the Renji Hospital affiliated to the Shanghai Second Medical University, were matched against 117 women with normal pregnancy history. Case-control study to compare the frequency of G/A alleles, AA/AG/GG genotypes and A + (AA + AG) /G+ (GG + AG) phenotypes of CTLA-4 between RSA patients and controls were performed. After amplification of CTLA-4 exon-1 region by polymerase chain reaction (PCR), restriction fragment-length polymorphism (RFLP) was used to detect the polymorphism at position 49 in exon-1 of CTLA-4 gene. Statistical significance was tested by SPSS software. RESULTS: There were dissimilar distributions of G/A alleles, AA/AG/GG genotypes and A+/G+ phenotypes of CTLA-4 between RSA patients and controls. The frequencies of G allele (P = 0.032) and GG genotype (P = 0.011) in RSA patients were significantly higher than those in controls, while the frequencies of AG genotype (P = 0.039) and A + (AA + AG) phenotype in RSA patients were decreased significantly (P = 0.011). CONCLUSIONS: Our findings suggest that A/G polymorphism in exon-1 of CTLA-4 is associated with the immunopathogenesis of RSA, and it confers susceptibility to RSA in Chinese population.     

QAP and QPB polymorphism in Mexican mestizos and Indians

Although the sequences of the class II promoters are highly conserved, diversity has been found in the URRs of DR and DQ loci. For the promoter region of DQA1, 10 QAP alleles are defined and 12 QBP for DQB1 region; DNA of 46 Mexican Mestizos and 101 Seri Indians was typed for QAP, QBP and class II alleles using the PCR-SSO protocols of the 12th W, as part of the promoter component chaired by E. Albert. PCR-SSP was done to distinguish between QBP6.2 and 6.3. In both groups, all QAP alleles previously described were detected, excepting for 3.2, absent because the associated DQA1^*0302 is also lacking. One unusual haplotype accounting for 16.3% was observed: DRB1 ^*0802-QAP4.2-DQA1^*0401-DQB1^*0402-QBP4.1. QBP3.22 described in Whites, was not present in this haplotype. Neither QBP4.1 that associates with DQB1 ^*0401, or the latter have been found in Mexicans. A recombination possibly occured in the DQB1 region of an ancestral haplotype carrying QBP4.1. In Seri Indians, only 8 haplotypes were detected. Of these, 3 seem to be the ancestral ones: ^*0407-^*03011-3.1-^*0302-3.21; ^*0802-4.2-^*0401-^*0402-4.1; and ^*1402-4.1-^*0501-^*0301-3.1. The frequencies of 43.5%, 36.6% and 13.9% accounting for 94% of the haplotypes, indicate from an evolutionary viewpoint, that these originated from Orientals and probably conferred a great biological advantage. They are also the prevalent haplotypes in Mestizos (60.9%). One unusual QBP4.1 combination was detected in 2 Seris with DRB1^*0407-DQB1^*0302. The functional role of these variants in expression and in development of disease, must be explored.     

TNF-α基因启动子多态性与多发性骨髓瘤易感性关系的观察

为了探讨TNF α基因启动区 30 8、 2 38位点多态与多发性骨髓瘤 (MM )易感性的关系。采用SSP PCR (sequencespecificprimer PCR )方法检测 5 6例中国汉族MM患者和 114名中国汉族健康对照者TNF α基因启动区 30 8、 2 38位点等位基因频率 ,分析两位点多态性与MM易感性的关系。结果显示 ,MM组与对照组 2 38位点A的频率分别为 15 18% (17/ 112 ) ;15 79%(36 / 2 2 8) ,两组间比较无显著差异 (P =0 884 ) ;MM组与对照组 30 8位点A的频率分别为 1 79% (2 / 112 ) ;9 2 1% (2 1/2 2 8) ,两组间比较有显著差异 (P =0 0 10 ) ,MM组明显低于对照组。结论 :在所研究的人群中未发现TNF α基因启动子 2 38位点多态性与MM易感性相关 ,但MM患者TNF α基因启动区 30 8位点A的频率明显低于对照组 ,推测TNF α 30 8位点A为MM发病的保护性因素     

Significant compatibility does not exist at the HLA-DQB gene locus in couples with unexplained recurrent abortions.

The polymerase chain reaction-restricted fragment length polymorphism (PCR-RFLP) method was used for both examining compatibility at the HLA-DQB1 gene locus and determining HLA-DQ antigen polymorphism in spouses of unexplained recurrent abortions. Genomic DNA samples were prepared from peripheral mononuclear cells from patient and control couples. Two hundred and thirty base pair fragments of the second exon of the HLA-DQB genes were selectively amplified. Amplified DNAs were digested with the restriction endonucleases, Fok I, Hae III, Hha I, Rsa I and Sau3A I, and subjected to electrophoresis in a polyacrylamide gel. The RFLPs showed that habitual aborters and their husbands had neither significantly frequent alleles nor shared common alleles at the HLA-DQB locus when compared to the control group. Since significant HLA-DQB compatibility was not observed between the spouses and unexplained recurrent aborters, in order to determine whether or not HLA compatibility is responsible for the genesis of unexplained recurrent abortions, it is imperative to further examine the compatibility between other HLA gene loci.