中国汉族群体c—Ha—ras基因3‘端—VNTR位点遗传多态性的初步研究

为了探讨中国汉族ｃ－Ｈａ－ｒａｓ基因３′端ＶＮＴＲ位点遗传多态性，用ＡＦＬＰ－ＰＡＧＥ法对１１２例无亲缘关系的健康个体进行了ｃ－Ｈａ－ｒａｓ基因３′ＶＮＴＲ位点分析，共检出２１个等位基因，其片段大小分布范围在６３５～２６５１ｂｐ之间，基因频率分布在０．００４５～０．５９３８，父权排除率（Ｐｅ）＝０．４４４５；杂合度（Ｈ）＝０．６２５，个体识别力（Ｄｐ）＝０．８４４０，共发现３４种基因型，经Ｘ＾２检     

与成人多囊肾病PKD1紧密连锁的四种微卫星DNA在中国人群中的多态性

目的为了评估成人多囊肾病ＰＫＤ１基因内的微卫星ＤＮＡＫＧ８及与ＰＫＤ１紧密连锁的微卫星ＳＭ６、ＣＷ４和ＣＷ２在基因诊断中的有效性。方法采用ＰＣＲ、聚丙烯酰胺凝胶电泳（ＰＡＧＥ）和银染法分析了部分无血缘关系的中国汉族人。结果在中国汉族人群中ＫＧ８有６种等位片段，其多态信息量（ＰＩＣ）为０．３１２；ＳＭ６具有２４种等位片段，其ＰＩＣ为０．８０；ＣＷ４有９种片段，ＰＩＣ为０．８５０；ＣＷ２有７种，ＰＩＣ为０．８１４。而白种人中ＫＧ８有８种，ＰＩＣ为０．５４５；ＳＭ６有１６种，ＰＩＣ为０．６５３；ＣＷ４有９种，ＰＩＣ为０．７８２；ＣＷ２有１３种，ＰＩＣ为０．８０９。结论研究人群与文献报道的白种人群相比，这四种微卫星在种类和分布上均有差异，表明二核苷酸重复在不同民族中存在差异；ＳＭ６、ＣＷ４和ＣＷ２是高度多态的遗传标记，可用于ＡＰＫＤ的连锁基因诊断、法医学上的个体鉴定和亲权鉴定     

与成人多囊肾病PKD1紧密连锁的四种微卫星DNA在中国人群中的？…

目的 为了评估成人多囊肾病ＰＫＤ１基因内的微卫星ＤＮＡＫＧ８及与ＰＫＤ１紧密连锁的微卫星ＳＭ６，ＣＷ４和ＣＷ２在基因诊断中的有效性。方法 采用ＰＣＲ，聚丙烯酰胺凝胶电泳（ＰＡＧＥ）和银染法分的了部分无血缘关系的中国汉族人。结果 在中国汉族人群中的ＫＧ８有６种等位片段，其多态性信息量（ＰＩＣ）为０．３１２；ＳＭ６具有２４种等位片段，其ＰＩＣ为０．８０；ＣＷ４有９种片段，ＰＩＣ为０．８５０，ＣＷ２有７     

Rb1基因位点P123M1．8多态性的建立及其在Wilson病中的连锁关系

视网膜母细胞瘤易感基因（Ｒｂ１）基因位点位于１３ｑ１４～２１，与Ｗｉｌｓｏｎ病基因位点相距４．４分摩（ｃｅｎｔｉｍｏｒｇａｎｓ，ｃＭ）．中国人Ｒｂ１基因５′端第１个内含子中Ｐ１２３Ｍ１，８标记的等位片段与白种人相同，但杂合率有差异。中国人Ｐ１２３Ｍ１．８／ＢａｍＨⅠ的多态性信息含量（ＰＩＣ）为０．５２，并与Ｗｉｌｓｏｎ病基因位点呈紧密连锁关系（θ＝０．０５，Ｚ＝３．８４６）。应用Ｐ１２３Ｍ１．８标记对９个Ｗｉｌｓｏｎ病家系进行连锁分析，证实该标记可用于Ｗｉｌｓｏｎ病的症状前诊断及杂合子检出。     

过敏性紫癜患者血浆GMP－140、cAMP、cGMP、IgG和IgA水平的研究

采用放射免疫法（ＧＭＰ－１４０用单位点免疫放射法）测定过敏性紫癜患者治疗前（ｎ＝２１）后（ｎ＝１９）血浆α－颗粒膜蛋白（ＧＭＰ－１４０）、环磷酸腺苷（ｃＡＭＰ）、环磷酸鸟苷（ｃＧＭＰ）、免疫球蛋白Ｇ和Ａ（ＩｇＧ、ＩｇＡ）含量，以３１例健康人作对照．结果，患者与对照比较，血浆ＧＭＰ－１４０（７５４±１６８分子数／血小板，Ｔ＝２．２９５，Ｐ＜０．０５），ｃＡＭＰ（１９．８９±７．９２对１４．２６±５．６３ｎｍｏｌ／Ｌ，Ｔ＝２．９９９，Ｐ＜０．０１），ｃＧＭＰ（４．８７±２．６２对３．４１±１．６９ｎｍｏｌ／Ｌ，Ｔ＝２．４４６，Ｐ＜０．０５），ＩｇＧ（１５．７５±５．５４对１１．４５±４．８６ｇ／Ｌ，Ｔ＝２．９５８，Ｐ＜０．０１）和ＩｇＡ（１．６８±０．８７对１．１６±０．５２ｇ／Ｌ，Ｔ＝２．６９８，Ｐ＜０．０１），且ｃＡＭＰ和ｃＧＭＰ水平升高呈正相关（ｒ＝０．４６９，Ｐ＜０．０５）。过敏性紫癜患者临床治愈后上述物质血浆含量下降，与对照比较无差异（Ｐ＞０．０５）．提示过敏性紫癜患者血浆ＧＭＰ－１４０、ｃＡＭＰ、ｃＧＭＰ、ＩｇＧ和ＩｇＡ可明显升高，临床治愈后恢复正常。     

P53融合蛋白的原核表达及其单克隆抗体的制备

目的 为研制出能特异与Ｐ＾５３结合的单克隆抗体,使对Ｐ＾５３的检测得到更为广泛的应用。方法 用ＰＣＲ技术扩增编码人Ｐ＾５３Ｎ－端１８０个氨基酸的ＤＮＡ片段并将其克隆在谷胱苷肽转移酶（ＧＳＴ）表达质粒Ｐ＾ＧＥＸ－２Ｔ中。用该重组质粒转化大肠杆菌ＪＭ１０９,并经异丙基β－Ｄ硫代半乳糖苷（ＩＰＴＧ）诱导产生Ｐ＾５３－ＧＳＴ融合蛋白。用Ｐ＾５３－ＧＳＴ免疫ＢＡＬＢ／ｃ小鼠。常规细胞融合,间接酶联免疫吸附试     

荷人鼻咽癌裸鼠全血GSH－PX和CAT活性的检测

用ＢＡＬＢ／ｃ裸小鼠（ｎｕｄｅｍｉｃｅ，ＮＭ）复制鼻咽癌（ｎａｓｏｐｈａｒｙｎｇｅａｌｃａｒｃｉｎｏｍａ，ＮＰＣ）模型，检测不同病期荷人ＮＰＣＮＭ全血谷胱甘肽过氧化物酶（ｇｌｕｔａｔｈｉｏｎｅｐｅｒｏｘｉｄａｓｅ，ＧＳＨ－ＰＸ）和过氧化氢酶（ｃａｔａｌａｓｅ，ＣＡＴ）活性，结果发现早期荷瘤鼠全血ＧＳＨ－ＰＸ活性明显高于正常裸鼠（Ｐ＜０．０１），然后随着病情发展有所下降，但中或晚期荷瘤鼠全血ＧＳＨ－ＰＸ活性仍高于正常水平（Ｐ＜０．０１）．而荷瘤鼠在潜伏、早和中期全血ＣＡＴ活性与正常裸鼠比较未见明显差异（Ｐ＞０．０５），仅在晚期荷瘤鼠全血ＣＡＴ活性明显低于正常裸鼠（Ｐ＜０．０１）．结果提示ＧＳＨ－ＰＸ和ＣＡＴ活性的改变与ＮＰＣ的发生和发展有关。     

与成人型多囊肾病基因位点1连锁的微卫星的遗传多态性研究

目的探讨广泛应用于高加索人多囊肾病（ＰＫＤ１）基因诊断的两个微卫星在中国汉族及壮族人群中的多态性及其等位基因频率是否存在群体差异。方法用聚合酶链反应（ＰＣＲ）扩增与ＰＫＤ１连锁的两个微卫星，１０％非变性聚丙烯酰胺凝胶电泳结合ＤＮＡ序列测定检测ＰＣＲ产物。结果ＡＣ２．５和ＳＭ７两个微卫星：（１）在汉族中分别观察到１０和１１个等位基因，期望杂合度分别为６８．４％及７７．４％，多态信息含量（ＰＩＣ）分别为０．６８及０．７７；（２）在壮族中分别观察到８及９个等位基因，期望杂合度分别６７．１％及５７．５％，ＰＩＣ分别为０．６７及０．５７；（３）６个ＡＣ２．５等位基因的频率及８个ＳＭ７等位基因的频率存在群体差异（Ｐ＜０．０５）。结论ＡＣ２．５和ＳＭ７在汉族和壮族中均高度多态（ＳＭ７在壮族中稍差），可用于汉、壮族人ＰＫＤ１基因诊断。同时，这两个微卫星的等位基因频率分布具有群体差异，在应用它们进行基因诊断和疾病关联性研究时应引起注意。     

Rb1基因位点P123M1.8多态性的建立及其在Wilson病中的…

视网膜母细胞瘤易感基因（Ｒｂ１）基因位点位于１３ｑ１４￣２１，与Ｗｉｌｓｏｎ病基因位点相距４．４分摩（ｃＭ）。中国人Ｒｂ１基因５＇端第１个内含子中Ｒ１２３Ｍ１．８标记的等位片段与白种人相同，但杂合率有差异。中国人Ｐ１２３Ｍ１．８／ＢａｍＨＩ的多态性信息含量（ＰＩＣ）为０．５２，并与Ｗｉｌｓｏｎ病基因位点呈紧密连锁关系（θ＝０．０５，Ｚ＝３．８４６）。应用Ｐ１２３Ｍ１．８标记对９个Ｗｉｌｏｓｎ病家     

超顺磁性氧化铁粒子（SPIO）的驰豫率及磁化率研究

本文采用Ｃａｒｒ－Ｐｕｒｅｌｌ－Ｍｅｉｂｏｏｍ－Ｇｉｌｌ序列及古埃及氏法分别测定了超磁显注射液与Ｅｎｄｏｒｅｍ的驰豫率和磁化率，结果表明：超磁显注射液的Ｔ２驰豫率与磁化率为０．１３８２９＊１０＾６ｍｏｌ＾－１．ｓｅｃ＾－１和３０３４６＊１０＾－５ｃｇｓ／ｇＦｅ与进口Ｅｎｄｏｒｅｍ的Ｔ２驰豫率及磁化率十分相近。     

Genome-wide analysis of microsatellite polymorphism in chicken circumventing the ascertainment bias

Studies of microsatellites evolution based on marker data almost inherently suffer from an ascertainment bias because there is selection for the most mutable and polymorphic loci during marker development. To circumvent this bias we took advantage of whole-genome shotgun sequence data from three unrelated chicken individuals that, when aligned to the genome reference sequence, give sequence information on two chromosomes from about one-fourth (375,000) of all microsatellite loci containing di- through pentanucleotide repeat motifs in the chicken genome. Polymorphism is seen at loci with as few as five repeat units, and the proportion of dimorphic loci then increases to 50% for sequences with approximately 10 repeat units, to reach a maximum of 75%-80% for sequences with 15 or more repeat units. For any given repeat length, polymorphism increases with decreasing GC content of repeat motifs for dinucleotides, nonhairpin-forming trinucleotides, and tetranucleotides. For trinucleotide repeats which are likely to form hairpin structures, polymorphism increases with increasing GC content, indicating that the relative stability of hairpins affects the rate of replication slippage. For any given repeat length, polymorphism is significantly lower for imperfect compared to perfect repeats and repeat interruptions occur in >15% of loci. However, interruptions are not randomly distributed within repeat arrays but are preferentially located toward the ends. There is negative correlation between microsatellite abundance and single nucleotide polymorphism (SNP) density, providing large-scale genomic support for the hypothesis that equilibrium microsatellite distributions are governed by a balance between rate of replication slippage and rate of point mutation.     

Norepinephrine Content in the Hypothalamus and Medulla Oblongata of ISIAH Rats is Regulated by Several Genetic Loci

Five genetic loci regulating epinephrine content in rat hypothalamus and two loci in the medulla oblongata were identified using polymorphic microsatellite DNA markers. Allele polymorphism of these loci determines the differences between norepinephrine levels in brain compartments of hypertensive ISIAH and normotensive WAG rats. The interactions between the detected epinephrine-regulating loci are additive.     

雷氏按蚊多态微卫星DNA位点的筛选和特征

本研究对雷氏按蚊的微卫星DNA序列进行了分离,并筛选了其中具有多态性的微卫星位点.应用雷氏按蚊基因组DNA的酶切片段与生物素标记的寡核苷酸探针杂交,亲合素富集和超滤离心浓缩目的片段,扩增放大后克隆并测序,获得雷氏按蚊微卫星DNA序列.在其中挑选合适的微卫星位点,建立扩增体系.应用雷氏按蚊现场样本对不同的微卫星DNA进行扩增,聚丙烯酰胺凝胶电泳筛选具有多态性的位点.本研究结果获得了雷氏按蚊微卫星DNA序列58条,GenBank注册登记号为EF620299～EF620356.其中,完整序列41条,占70.7%;非完整序列7条,占12.1%,复合序列9条,占15.7%;仅1条为非典型序列.电泳结果显示其中14个微卫星位点中有11个具有多态性.     

Association of polymorphic MHC microsatellites with GVHD, survival, and leukemia relapse in unrelated hematopoietic stem cell transplant donor/recipient pairs matched at five HLA loci

In order to determine whether matching/mismatching for microsatellite polymorphism provides useful information on acute graft-vs-host disease (GVHD), survival, and leukemia relapse in hematopoietic stem cell (HSC) transplantation, we genotyped for polymorphisms at 13 microsatellite loci within the major histocompatibility complex (MHC) of 100 unrelated HSC transplant donor-recipient pairs who were matched at five classical human leukocyte antigen (HLA) loci. A high percentage of allele matching was obtained for five microsatellite loci, DQCARII (96%), MICA (93%), MIB (89%), C1-3-1 (93%), and D6S510 (97%), that are localized within 100 kb of the HLA-DR, HLA-DQ, HLA-B, HLA-C, or HLA-A locus. In contrast, the other eight microsatellites are located farther away from the HLA classical loci and have much lower percentages of allele matching [e.g. tumor necrosis factor a (TNFa) (73%), TNFd (74%), D6S273 (64%), C3-2-11 (46%), C5-3-1 (50%), C5-4-5 (63%), C5-2-7 (68%), and D6S265 (81%)]. Therefore, there were at least eight microsatellite markers with relatively high percentages of mismatches in the donor/recipient pairs with acute or chronic GVHD, poor graft survival, and leukemia relapse. However, there were no statistically significant associations between mismatched donor-recipient pairs at the 13 microsatellite loci and acute or chronic GVHD, graft survival, and leukemia relapse. Nevertheless, allele matching at the microsatellite TNFd locus near the TNFa gene was found by the Fisher's exact double-sided test to be significantly associated with decreased survival in the grade III/IV acute GVHD group. Overall, these results suggest that the matching of microsatellite polymorphisms within the HLA region, especially the ones farthest from the classical HLA loci, was not useful indicator for the outcome of HSC transplantation from unrelated donors. In this regard, the future determination of the genome-wide microsatellite genotypes in HLA-matched donor-recipient pairs, outside the MHC, may be a better possibility for identifying minor histocompatibility genes in linkage disequilibria with microsatellites as potential predictive markers for the occurrence of acute GVHD and survival rate in HSC transplantation.     

上海汉族人群MICA基因第5外显子微卫星多态性研究

目的 了解上海地区汉族人群ＭＩＣＡ基因第５外显子微卫生多态性分布,以及ＭＩＣＡ基因与其紧密连锁的ＨＬＡ－Ｂ基因位点的关系。方法 用ＰＣＲ－异源双链分析法,对１７５名正常无关个体的ＭＩＣＡ基因第５外显子微卫星多态性的分布进行研究。结果 （１）上海地区汉族人群ＭＩＣＡ基因第５外显子存在５种等位基因,其中ＭＩＣＡ＊Ａ５最为常见（３９．１４％）,其次为＊Ａ５．１（２２．２９％）;（２）ＭＩＣＡ等位基因与Ｈ     

非小细胞肺癌6号染色体长臂的基因扫描分析

目的 探讨6号染色体长臂上与非小细胞肺癌发生相关的微卫星位点。方法 应用多重PCR对41例非小细胞肺癌中6号染色体长臂上的18个微卫星位点进行扩增。PCR产物应用聚丙烯酰胺凝胶电泳分离，电泳结果用Gene Scam^TM,Genotyper^TM软件进行分析，结果 各个位点有明显的不同的杂合性缺失（loss of heterozygosity,LOH）频率，从3.85%到38.45%不等，总LOH频率为58.5%(24/41)。LOH频率超过20％的位点共有8个，主要分布的6q24及6q27。具体范围为6q24-6q25.3[D6S1699(35%)、D6S409(23.33%)、D6S441(33.33%)]及6q26-27[D6S1550(38.45%)、D6S264(20%)、D6S1585(25%)、D6S446(33.33%)、D6S281(30.77%]。结论 6q24及6q27附近可能存在与非小细胞肺癌发生相关的肿瘤抑制基因。     

Twenty-six new polymorphic microsatellite markers around the HLA-B, -C and -E loci in the human MHC class I region

Abstract: The human major histocompatibility complex (MHC) class I region is believed to contain a large number of disease-related loci for diseases such as Behçet's disease and psoriasis vulgaris. Although many novel genes have recently been identified in this region, it still appears to be difficult to relate any of these new genes to MHC class I-associated diseases as causative genetic factors. During the course of large-scale genomic sequencing of the human MHC class I region, we identified 262 microsatellite sequences with dinucleotide to pentanucleotide repeats around the HLA-B, -C and HLA-E genes. Of these, 26 microsatellites were investigated for repeat polymorphism using 60 HLA homozygous B-cell lines and 60 healthy random individuals. The average number of alleles at these microsatellite loci was 9.6 with a PIC (polymorphism content value) of 0.69. These new polymorphic microsatellite markers will probably be very useful for precise mapping of disease-related genes within the HLA class I region in linkage analysis. Moreover, they will provide a powerful tool to study recombination events in this region, which contributes to haplotypic diversification.     

应用微卫星DNA基因分型技术进行双生子卵型鉴定

目的 用微卫星DNA基因扫描和分型技术建立双生子卵型鉴定法。方法 选取69对同性别双生子、6对异性别双生子和17对同胞对,抽提基因组DNA,单盲设计,随机编号后,采用9对荧光标记的,在中国人群中具有高度杂合度的短串联重复序列（short tandem repeat,STR)引物,进行基因扫描和分型分析。根据这9个STR基因型的一致性来鉴别卵型。结果 9个STR基因型完全一致的63对受检者全部为同性别双生子,6对异性别双生子和17对同胞对的STR基因型均不完全一致,另有6对同性别双生子的STR基因型也不完全一致,经计算采用6个或5个STR位点判定同性别同卵双生的可信性分别大于99.6%和99%,采用全部9个位点判定同性别同卵双生的可信性大于99.95%。结论 STR基因扫描和分型技术为在基因组水平上直接判别双生子卵型,提供了一种准确,可靠的鉴定方法,它还有快速,简便等优点。     

具有家族史鼻咽癌4p15.1-4q12区等位基因杂合性丢失分析

目的确定有家族史鼻咽癌患者4p15．1-4q12区域等位基因杂合性丢失（loss of heterozygosity，LOH）的分布和频率，为进一步缩小该区域内易感基因的范围提供新的线索和依据。方法收集具有家族史的鼻咽癌患者石蜡包埋的活检组织标本，采用显微切割的方法在肿瘤组织石蜡切片上分离肿瘤细胞和正常淋巴细胞，选定7个定位于4p15．1-4q12区域内的高密度微卫星位点，多重PCR扩增和丙烯酰胺凝胶电泳后，Genescan软件对各位点LOH进行分析。结果25例具家族史鼻咽癌患者中，23例在4p15．1-4q12区至少存在一个微卫星位点的LOH（92％）。其中D4S2382位点LOH的频率最高，达到56％；D4S350和D4S1547位点LOH频率均约为50％。最小共同缺失区位于位点D4S350和D4S1547之间。结论鼻咽癌4p15．1—4q12区域内的易感基因可能位于微卫星位点D4S350和D4S1547附近。     

广西壮族人群13个单核苷酸基因座遗传多态性

目的 采用荧光标记复合扩增单核苷酸多态性(single nucleotide polymorphism,SNP)体系对广西地区壮族314名个体的13个SNP基因座进行等位基因频率调查,并评价其法医学应用价值.方法 选择13个常染色体双等位基因SNP基因座,应用荧光标记片段长度差异等位基因特异性复合扩增SNP分型体系,对广西地区壮族人群进行群体调查.结果 获得了广西地区壮族人群13个SNP基因座的等位基因频率,13个SNP基因座等位基因频率分布均符合Hardy-Weinberg平衡,杂合度位于0.2166和0.5478之间,多态信息总量位于0.2084与0.3750之间,13个SNP基因座累积个人识别几率和累积非父排除率均分别达到99.99%和87.71%.结论 用荧光标记复合扩增片段长度差异等位基因特异性PCR法可同时对多个SNP基因座进行分型;13个SNP基因座在法医学个人识别领域具有较高的应用价值.