HLA-B27基因在云南傣族健康人群和云南汉族白血病的研究
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摘要
目的:①运用序列特异性引物聚合酶链反应(polymerase chain reaction-sequence specific primers,PCR-SSP)扩增方法检测HLA-B27基因在云南西双版纳傣族健康人群中的分布,以了解HLA-B27基因在云南西双版纳傣族人群中的分布特征,并为种族遗传学提供重要的参考资料。
②运用序列特异性引物聚合酶链反应(polymerase chain reaction-sequencespecific primers,PCR-SSP)扩增方法检测HLA-B27基因与云南汉族白血病患者的关联性,为白血病的发病机制提供一些遗传学参考依据,并为下一步进行HLA与云南不同民族白血病的相关性研究提供基础数据。
方法:①根据“知情同意”的原则,在云南西双版纳傣族自治州景洪市傣族人群聚居地,选择3代以上相互之间无亲缘关系,均为本民族的健康个体61例。本研究首先合成扩增HLA-B基因的引物一对,扩增HLA-B27基因的引物两对,然后建立PCR-SSP方法。从EDTA抗凝的外周血提取基因组DNA,然后对HLA-B27基因进行PCR扩增。运用四格表X~2检验分析HLA-B27基因在云南傣族健康人群和云南汉族、纳西族、傈僳族、怒族、彝族、中国南、北方汉族及泰国健康人群中的分布差异。
②随机收集2007年5月-11月在昆明医学院第二附属医院血液科住院,确诊为白血病的患者26例作为病人组,健康对照组选取62例来自云南昆明汉族无血缘关系的健康者。本研究首先合成扩增HLA-B基因的引物一对,扩增HLA-B27基因的引物两对,然后建立PCR-SSP方法。从EDTA抗凝的外周血提取基因组DNA,然后对HLA-B27基因进行PCR扩增。按照病例-对照研究分析方法,分析HLA-B27基因在白血病组与健康对照组阳性率的差异及HLA-B27基因与疾病的相关性。
结果:①在关于61例云南傣族健康人群的HLA-B27基因研究中,14例为阳性,占22.9%,明显高于云南汉族(4.8%)、纳西族(0.6%)、傈僳族(0.9%)、彝族(4.2%)、怒族(0),亦明显高于中国南方汉族(4.3%)、北方汉族(4.29%)、泰国人群(10%),经四格表X~2检验分析,HLA-B27基因在云南傣族健康人群与云南汉族、纳西族、傈僳族、怒族、彝族、中国南、北方汉族及泰国人群中基因阳性率具有显著性差异。
②在关于26例云南汉族白血病患者的HLA-B27基因研究中,发现1例为阳性,占3.8%。在62例云南汉族健康人群中,有3例为阳性,占4.8%。按照病例-对照研究分析方法,经四格表X~2检验分析,无统计学差异。
结论:①在云南傣族健康人群的HLA-B27基因研究中,发现HLA-B27基因在云南傣族健康人群中的分布明显高于云南汉族、纳西族、傈僳族、怒族、彝族,亦明显高于中国南、北方汉族及泰国人群。本研究结果表明,西双版纳傣族是一个与汉族群体及其他群体不同的、具有HLA-B27基因高频率的群体。可能是由于不同区域、不同种族及人群HLA分布本身就存在很大的差异,也可能是由于不同人群的生存环境、生活习惯及实验方法不同等因素的影响,造成HLA-B27基因阳性率分布的不均衡,因此需要尽可能的排除影响因素,并进一步收集样本减少误差。
②在本组云南汉族白血病与HLA-B27基因的关联性研究中,没有发现HLA-B27基因与云南汉族白血病具有相关性,也没有发现HLA-B27基因在本组白血病患者与正常健康人中的分布有差异性,这可能与白血病的复杂的发病机制,分类较多及人群地理分布及样本量有关。需要进一步收集大量样本,对不同民族、不同类型的白血病进行与HLA关联性的研究。
Objective:①To understand the distribution of HLA-B27 gene in healthy population from Yunnan Dai Ethnic Group,by polymerase chain reaction-sequence specific primers(PCR-SSP),HLA-B27 gene was amplified from the healthy Dai inhabiting in Xishuangbanna Autonomous Prefecture in Yunnan Province,that will provide important references for racial genetics.
②Using polymerase chain reaction-sequence specific primers to amplify HLA-B27 gene in patients with leukemia in Yunnan Han people and study the association of HLA-B27 gene with these patients,that provide some reference in genetics for the pathogenesis of leukemia and provide the basic data for the relevance of HLA to different ethnic groups with leukemia in Yunnan for study of the next step.
Methods:①A total of 61 healthy unrelated individuals from Dai ethnic groups lived in Xishuangbanna Autonomous Prefecture in yunnan province of southwestern China were enrolled in this study.The peripheral blood was collected after informed consents were obtained,and then the HLA-B27 gene was amplified by PCR -SSP.Using four-frame Table X~2 examination to analysis differences of HLA-B27 gene frequencies in healthy unrelated individuals from Yunnan Dai and Han、Naxi、Lisu、Nu、Yi、the South Han、the North Han in China and Thai people in Thailand.②26 patients with leukemia were diagnosised in the second hospital of Kunming Medical College,between in May 2007 and November 2007,who were selected randomly from the department of Hematology.62 Yunnan Han Who lived in Yunnan province of southwestern China were enrolled as the control.After extrating the genomic DNA from peripheral blood anticoagulanted by EDTA and then the HLA-27 gene was amplified by PCR-SSP.According to case-control study analysis,we analysised the difference of HLA-B27 gene frequencies between in the healthy control group and the Leukemia Group and studied the association of HLA-B27 gene with patients with leukemia.
Result:①61 cases in Yunnan Dai healthy population for HLA-B27 gene,14 cases were positive,accounting for 22.9%,significantly higher than that of Yunnan Han(4.8%)、Naxi(0.6%)、Lisu(0.9%)、Nu(0%)、Yi(4.2%),China Southern Han(4.3%)、the North Han(4.29%)and Thai people(10%).Using the four-frame table X~2 test, studied the distribution of HLA-B27 gene frequencies among Yunnan Dai、Han、Naxi、Lisu、Nu、Yi、Southern Han、North Han in China and Thai people.There are significantly different to frequencies of HLA-B27 gene among these people.
②In 26 cases of patients with leukemia in Yunnan about HLA-B27gene research, we found one cases is positive,for 3.8 percent,and in 62 cases Han people,three cases is positive,accounting for 4.8%.According to case-control study on the analysis method,there was no statistically different between the patients with leukemia and the control subject.
Conclusion:①In the study about,we found that frequence of HLA-B27 gene in healthy people from Yunnan Dai ethnic group was significantly higher than that of the Han、Naxi、Lisu、Nu、Yi、and also significantly higher than Southern Han、the North Han in China and Thai people in Thailand.The results of this study show that, Xishuangbanna Dai Han is a different group with a high frequency of HLA-B27 gene comparing with other groups.May be due to different regions,different ethnic groups and the distribution of HLA itself that there is a big difference,or be due to living environment where different groups of people lived,living habits of different groups of people and the different experimental methods,and so on,that result in the distribution of HLA-B27 gene frequency uneven.Hence need to exclude affecting factors as much as possible and need to collect a larger sample and go to further reduce the error.
②In the study of this group to the relevance of HLA-B27 gene and the patients with leukemia in Yunnan Han,we didn't find that there were the relevance of HLA-B27 gene and the patients with leukemia,and also didn't find that there were difference between in the patients with leukemia and in healthy people to the distribution of HLA-B27 gene frequencies.This may be related to the incidence of leukemia with complicated mechanism,more classification and geographical distribution of the samples,sample size,and so on.Need to collect large numbers of samples and study association of different types of leukemia from different nationalities with HLA.
②运用序列特异性引物聚合酶链反应(polymerase chain reaction-sequencespecific primers,PCR-SSP)扩增方法检测HLA-B27基因与云南汉族白血病患者的关联性,为白血病的发病机制提供一些遗传学参考依据,并为下一步进行HLA与云南不同民族白血病的相关性研究提供基础数据。
方法:①根据“知情同意”的原则,在云南西双版纳傣族自治州景洪市傣族人群聚居地,选择3代以上相互之间无亲缘关系,均为本民族的健康个体61例。本研究首先合成扩增HLA-B基因的引物一对,扩增HLA-B27基因的引物两对,然后建立PCR-SSP方法。从EDTA抗凝的外周血提取基因组DNA,然后对HLA-B27基因进行PCR扩增。运用四格表X~2检验分析HLA-B27基因在云南傣族健康人群和云南汉族、纳西族、傈僳族、怒族、彝族、中国南、北方汉族及泰国健康人群中的分布差异。
②随机收集2007年5月-11月在昆明医学院第二附属医院血液科住院,确诊为白血病的患者26例作为病人组,健康对照组选取62例来自云南昆明汉族无血缘关系的健康者。本研究首先合成扩增HLA-B基因的引物一对,扩增HLA-B27基因的引物两对,然后建立PCR-SSP方法。从EDTA抗凝的外周血提取基因组DNA,然后对HLA-B27基因进行PCR扩增。按照病例-对照研究分析方法,分析HLA-B27基因在白血病组与健康对照组阳性率的差异及HLA-B27基因与疾病的相关性。
结果:①在关于61例云南傣族健康人群的HLA-B27基因研究中,14例为阳性,占22.9%,明显高于云南汉族(4.8%)、纳西族(0.6%)、傈僳族(0.9%)、彝族(4.2%)、怒族(0),亦明显高于中国南方汉族(4.3%)、北方汉族(4.29%)、泰国人群(10%),经四格表X~2检验分析,HLA-B27基因在云南傣族健康人群与云南汉族、纳西族、傈僳族、怒族、彝族、中国南、北方汉族及泰国人群中基因阳性率具有显著性差异。
②在关于26例云南汉族白血病患者的HLA-B27基因研究中,发现1例为阳性,占3.8%。在62例云南汉族健康人群中,有3例为阳性,占4.8%。按照病例-对照研究分析方法,经四格表X~2检验分析,无统计学差异。
结论:①在云南傣族健康人群的HLA-B27基因研究中,发现HLA-B27基因在云南傣族健康人群中的分布明显高于云南汉族、纳西族、傈僳族、怒族、彝族,亦明显高于中国南、北方汉族及泰国人群。本研究结果表明,西双版纳傣族是一个与汉族群体及其他群体不同的、具有HLA-B27基因高频率的群体。可能是由于不同区域、不同种族及人群HLA分布本身就存在很大的差异,也可能是由于不同人群的生存环境、生活习惯及实验方法不同等因素的影响,造成HLA-B27基因阳性率分布的不均衡,因此需要尽可能的排除影响因素,并进一步收集样本减少误差。
②在本组云南汉族白血病与HLA-B27基因的关联性研究中,没有发现HLA-B27基因与云南汉族白血病具有相关性,也没有发现HLA-B27基因在本组白血病患者与正常健康人中的分布有差异性,这可能与白血病的复杂的发病机制,分类较多及人群地理分布及样本量有关。需要进一步收集大量样本,对不同民族、不同类型的白血病进行与HLA关联性的研究。
Objective:①To understand the distribution of HLA-B27 gene in healthy population from Yunnan Dai Ethnic Group,by polymerase chain reaction-sequence specific primers(PCR-SSP),HLA-B27 gene was amplified from the healthy Dai inhabiting in Xishuangbanna Autonomous Prefecture in Yunnan Province,that will provide important references for racial genetics.
②Using polymerase chain reaction-sequence specific primers to amplify HLA-B27 gene in patients with leukemia in Yunnan Han people and study the association of HLA-B27 gene with these patients,that provide some reference in genetics for the pathogenesis of leukemia and provide the basic data for the relevance of HLA to different ethnic groups with leukemia in Yunnan for study of the next step.
Methods:①A total of 61 healthy unrelated individuals from Dai ethnic groups lived in Xishuangbanna Autonomous Prefecture in yunnan province of southwestern China were enrolled in this study.The peripheral blood was collected after informed consents were obtained,and then the HLA-B27 gene was amplified by PCR -SSP.Using four-frame Table X~2 examination to analysis differences of HLA-B27 gene frequencies in healthy unrelated individuals from Yunnan Dai and Han、Naxi、Lisu、Nu、Yi、the South Han、the North Han in China and Thai people in Thailand.②26 patients with leukemia were diagnosised in the second hospital of Kunming Medical College,between in May 2007 and November 2007,who were selected randomly from the department of Hematology.62 Yunnan Han Who lived in Yunnan province of southwestern China were enrolled as the control.After extrating the genomic DNA from peripheral blood anticoagulanted by EDTA and then the HLA-27 gene was amplified by PCR-SSP.According to case-control study analysis,we analysised the difference of HLA-B27 gene frequencies between in the healthy control group and the Leukemia Group and studied the association of HLA-B27 gene with patients with leukemia.
Result:①61 cases in Yunnan Dai healthy population for HLA-B27 gene,14 cases were positive,accounting for 22.9%,significantly higher than that of Yunnan Han(4.8%)、Naxi(0.6%)、Lisu(0.9%)、Nu(0%)、Yi(4.2%),China Southern Han(4.3%)、the North Han(4.29%)and Thai people(10%).Using the four-frame table X~2 test, studied the distribution of HLA-B27 gene frequencies among Yunnan Dai、Han、Naxi、Lisu、Nu、Yi、Southern Han、North Han in China and Thai people.There are significantly different to frequencies of HLA-B27 gene among these people.
②In 26 cases of patients with leukemia in Yunnan about HLA-B27gene research, we found one cases is positive,for 3.8 percent,and in 62 cases Han people,three cases is positive,accounting for 4.8%.According to case-control study on the analysis method,there was no statistically different between the patients with leukemia and the control subject.
Conclusion:①In the study about,we found that frequence of HLA-B27 gene in healthy people from Yunnan Dai ethnic group was significantly higher than that of the Han、Naxi、Lisu、Nu、Yi、and also significantly higher than Southern Han、the North Han in China and Thai people in Thailand.The results of this study show that, Xishuangbanna Dai Han is a different group with a high frequency of HLA-B27 gene comparing with other groups.May be due to different regions,different ethnic groups and the distribution of HLA itself that there is a big difference,or be due to living environment where different groups of people lived,living habits of different groups of people and the different experimental methods,and so on,that result in the distribution of HLA-B27 gene frequency uneven.Hence need to exclude affecting factors as much as possible and need to collect a larger sample and go to further reduce the error.
②In the study of this group to the relevance of HLA-B27 gene and the patients with leukemia in Yunnan Han,we didn't find that there were the relevance of HLA-B27 gene and the patients with leukemia,and also didn't find that there were difference between in the patients with leukemia and in healthy people to the distribution of HLA-B27 gene frequencies.This may be related to the incidence of leukemia with complicated mechanism,more classification and geographical distribution of the samples,sample size,and so on.Need to collect large numbers of samples and study association of different types of leukemia from different nationalities with HLA.
引文
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[1]Marsh SGE,Albert ED,Bodmer WF etal.Nomenclature for factors of the HLA system 2002[J].Eur J Immunogenet 2002,29:463-515.
[2]Schreuder GMT,Hurley CK,Marsh SGE etal.The HLA dicyionary 2004:a summary of HLA-A,-B,-C,-DR and -DQB1 alleles and their association with serologically defined HLA-A、-B,-C,-DR and-DQ antigens[J].Int J Immunogenet 2005,32:19-69.
[3]ThudeH,Hardt C,Schorner U,Ferencik S,Helfricht,Barz D.Identification of a new HLA-B allele,HLA-B*4443,in a German family[J].Tissue Antigens 2005,66:696-9.
[4]Robinson J,Waller MJ,Parham P,de Groot N,Bontrop R,etal.IMGT/HLA and IMGT/MH -C:sequence databases for thestudy of the major histoco mpatibility complex[J].Nucleic Acids Research,2003,31(1):31
[5]沈关心 主编.医学免疫学,第二版[M].人民卫生出版社,2007:74-76
[6]邓建强.HLA和疾病的相关性.西安医科大学学报[J].1998,19:650-652
[7]Klitz W.Immunogenetics.Viruses,cancer and the MHC[J].Nature,1992,356:17-18
[8]戴云鹏,阎文英,陈力军等.小儿急性淋巴细胞白血病HLA-A、B、D等位基因多态性研究[J].白血病·淋巴瘤,2006,15:330-332
[9]段渊,胡亚美,崔建涛等,HLA抗原与儿童急性白血病的相关性研究[J].中华血液学杂志.2000,8:21-22.
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[12]何 军,陈子兴,姚 利等,HLA基因多态性与慢性粒细胞和急性淋巴细胞白血病易感性的关联分析[J].基础医学与临床,2006,11:1260-02
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[1]Marsh SGE,Albert ED,Bodmer WF etal.Nomenclature for factors of the HLA system 2002[J].Eur J Immunogenet 2002,29:463-515.
[2]Schreuder GMT,Hurley CK,Marsh SGE etal.The HLA dicyionary 2004:a summary of HLA-A,-B,-C,-DR and -DQB1 alleles and their association with serologically defined HLA-A、-B,-C,-DR and-DQ antigens[J].Int J Immunogenet 2005,32:19-69.
[3]ThudeH,Hardt C,Schorner U,Ferencik S,Helfricht,Barz D.Identification of a new HLA-B allele,HLA-B*4443,in a German family[J].Tissue Antigens 2005,66:696-9.
[4]Robinson J,Waller MJ,Parham P,de Groot N,Bontrop R,etal.IMGT/HLA and IMGT/MH -C:sequence databases for thestudy of the major histoco mpatibility complex[J].Nucleic Acids Research,2003,31(1):31
[5]沈关心 主编.医学免疫学,第二版[M].人民卫生出版社,2007:74-76
[6]邓建强.HLA和疾病的相关性.西安医科大学学报[J].1998,19:650-652
[7]Klitz W.Immunogenetics.Viruses,cancer and the MHC[J].Nature,1992,356:17-18
[8]戴云鹏,阎文英,陈力军等.小儿急性淋巴细胞白血病HLA-A、B、D等位基因多态性研究[J].白血病·淋巴瘤,2006,15:330-332
[9]段渊,胡亚美,崔建涛等,HLA抗原与儿童急性白血病的相关性研究[J].中华血液学杂志.2000,8:21-22.
[10]梁晓岚,杨丛林,万长春,等.HLA基因与成人急性淋巴细胞白血病的相关性研究[J].白血病·淋巴瘤,2004,13(1):17-19.
[11]高素青,邓志辉,金士正等。中国南方汉族急性淋巴细胞白血病与HLA基因相关性研究[J]。中国实验血液学杂志,2005,13(2):210-214
[12]何 军,陈子兴,姚 利等,HLA基因多态性与慢性粒细胞和急性淋巴细胞白血病易感性的关联分析[J].基础医学与临床,2006,11:1260-02
[13]BortinMM,D'Amaro J,Bach FH,et al.HLA associations with leukemia[J].Blood,1987,70:227-232
[14]Linet MS.Bias W B,Dorgan JF,etal.HLA antigens in chronic lymphocytic leukemia[J].Tissue Antigens,1988,31:71-78.
[15]谭建明,周永昌,唐孝达,组织配型技术与临床应用[M],北京:人民卫生出版社,2002,521-522.
[16]Hester JP,Rossen R,Trujillo J,et al.Frequency of HLA antigens in chronic myelocytic leukemia [J].SouthMed J,1977;70:691-693
[17]Ruuska P,Silvennoinen Kassinen S,Timonen T,et al.Chronic mye-locytic leukemia:HLA association and decreased erythrocyte C3b receptor expression[J].Clin Exp Immunol,1985,59:300
[18]Posthuma EF,Falkenburg JH,Apperley JF,et al.HLA-B8 and HLA-A3 coexp ressed with HLA-B8 are associated with a reduced risk of development of chronic myeloid leuKemia[J].Blood,1999,93:3863- 3865.
[19]Posthuma EFM,Falkenburg JHF,Apperley JF,etaI.HLA-DR4 is associated with a diminished risk of the development of chronic myeloid leukemia(CML)[J].Leukemia,2000,14:859-862.
[20]曹孟德,秦东春,岳保红等.HLA-DRBl基因多态性与急性淋巴细胞白血病和慢性髓性白血病易感性关联的研究[J].中华微生物和免疫学杂志,2001,21(3):278-280.
[21]Fatma D,Sevgi D,etal.HLA System Affects the Age-at-Onset in Chronic Myeloid Leukemia [J].Am.J.Hematol.2003.73:256-262
[22]Cortes J,Fayad L,Kantarjian H,etal..Association of HLA phenotype and response to interferon -alpha in patients with chronic myelogenous leukemia[J].Leukemia,1998,12(4):455-462
[23]李丹,郄斌,于燕等.HLA与慢性粒细胞白血病相关性的调查[J].中国输血杂志,2001,8:14-4.
[24]周 丹,邹红岩,金士正等.南方汉族人群白血病患者与HLA基因相关性的研究[J].江西医学检验,2005,23:316-319.
[25]李丹,郗斌,刘海燕,于燕.HLA-I类基因与白血病相关性研究[J],中国实验血液学杂志,2005,(13)4:563-566
[26]戴云鹏,阎文英,马秀峰等.山东省急性淋巴细胞白血病患儿HLA-A、B、DRBl等位基因多态性研究[J].山东大学学报·医学版,2003,41:288-291
[27]王燕,郭玉红.HLA-DRBl基因多态性与儿童急性淋巴细胞白血病易感性的关联[J].山西医科大学学报,2002,27:220-221
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[30]Amiel JI..Study of the leukocyte Phenotypes in Hodgkin's disease Histocompatibility Testing.Baltimore[J].Williams and Wikins.1967.79
[31]孙步彤,顾春梅,关 丽等.非霍奇金淋巴瘤HLA表达的研究[J].白求恩医科大学学报,1999,25:407-409
[32]Peter Reimer,Wolfgang Fischbach.,etal.Decreased frequency of HLA-B35 in patients with gastric MALT lymphoma[J].Ann Hematol,2004,83:232-236
[33]Hee Baeg Choi,Sang-Young Roh,etal.Association of HLA alleles with non-Hodgkin's lymphoma In Korean population[J].Int J Hematol,2008,87:203-209
[34]邵文硕,再生障碍性贫血与HLA[J]。医学综述,1998,4:420-422
[35]Gluckman E,PaponL,HorsJ,etal.HLA markers in patients suffering from aplastican aemia[J].Haematologia,1981,14:165-172
[36]殷石,马文成.再障性贫血与HLA相关性研究[J].黑龙江医学,1996,9:6-7
[37]杨华,孙明艳,张伟杰等.再生障碍性贫血与HLA相关性研究[J],哈尔滨医药,1997,17:11-12
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