我国汉族儿童维生素D受体基因多态性分布

目的　了解我国汉族儿童维生素D受体 (VDR)基因多态性分布。方法　利用限制性内切酶BsmⅠ ,采用聚合酶链反应限制性片段长度多态性技术 (PCR -RFLP) ,对 1 69名健康汉族儿童进行了VDR基因分型 ,并计算其基因型频率分布。结果　中国汉族儿童VDR基因bb、Bb、BB基因型分别为 91 1 2 %、7 1 %、1 77%,明显不同于高加索人种 ,与韩国人较为相似。结论　VDR基因多态性具有种族差异性     

维生素D受体基因多态性与乳腺癌的相关性

目的：研究维生素D受体基因的多态性与乳腺癌的关系。方法：收集86例乳腺癌患者及134名对照,用聚合酶链反应－限制性片段长度多态性方法,在维生素D受体基因的3’端分析了两个限制性酶切位点（ApaI及TaqI）的多态分布。结果：发现TaqI位点等位基因在两个群体间分布的差异有显著性（P＝0．0004）。进一步对基因型进行分析发现,Tt、tt基因型与乳腺癌相关。而ApaI位点两等位基因未发现在两群体中存在差异。对ApaI及TaqI两座位的单体型进行分析,发现tA间存在连锁不平衡。在两群体中分析单体型的分布发现tA在病例中的比例明显高于对照人群,提示tA单体型与乳腺癌相关。两个位点等位基因及单体型与临床指标的分析均未发现阳性结果。结论：维生素D受体基因的多态怀与乳腺癌有关,提示维生素D受体基因与乳腺癌有关。     

维生素D受体基因多态性与骨关节炎关系的Meta分析

背景:维生素D受体与骨关节炎存在密切联系,而维生素D受体基因多态性被认为能够调控维生素D受体,进而影响骨关节炎的发生,但现有的研究仍存在争议。目的:确认维生素D受体基因多态性与骨关节炎的关系。方法:系统检索PubMed、Web of Science、EMbase、Cochrane Library、中国生物医学文献数据库、中国学术期刊全文数据库、万方、维普等数据库,检索时限均为建库截至2019年10月,检索所有提供了骨关节炎患者和非骨关节炎患者维生素D受体基因(ApaI、Bsm I、TaqI和Fok I)多态性数据的病例对照研究,采用Stata 14.0统计软件进行分析。结果与结论:①总共纳入21篇相关研究,共7 109例患者,其中包括骨关节炎患者3 123例,非骨关节炎患者4 006例;②Meta分析显示,欧洲人维生素D受体Bsm I多态性与骨关节炎之间存在相关性[(BB vs. bb:OR=1.677,95%CI(1.051,2.676),P=0.030;BB vs. Bb+bb:OR=1.780,95%CI(1.175,2.697),P=0.007],但只有3篇研究;亚洲人维生素D受体FokI多态性与骨关节炎之间存在相关性[(FF vs. Ff+ff:OR=0.609,95%CI(0.410,0.907),P=0.015],只有3篇研究;维生素D受体TaqI和ApaI多态性与骨关节炎无显著相关性,在排除异质性后结果也无明显相关性;③结果表明,维生素D受体ApaI、Bsm I、TaqI和FokI多态性可能与骨关节炎无明显相关性。     

维生素D受体Fok Ⅰ基因多态性对SLE患者维生素D受体表达的影响

目的:检测维生素D受体(VDR)FokⅠ基因多态性与系统性红斑狼疮(SLE)的相关性,以及对SLE患者VDR mRNA表达的影响。方法:应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)分析技术检测VDRFokI基因位点及基因型在271例SLE患者和130例健康人中的分布情况;并应用实时定量聚合酶链反应(RT-PCR)技术检测VDR mRNA在48例SLE患者和38例健康对照组的表达。结果:VDRFokⅠ多态性等位基因F和f频率在SLE组和健康对照组有统计学差异(P=0.001),携带F等位基因个体发生SLE的相对危险度(OR)为1.630(95%CI=1.210-2.196,P=0.001);FF纯合子基因型频率在SLE组也明显高于健康对照组(42.8%vs25.4%,χ2=11.417,P=0.001)。同时,携带F等位基因的SLE患者(FF基因型和Ff基因型患者),浆膜炎发生率(P=0.001)及抗ds-DNA抗体(P=0.001)、抗Sm抗体(P=0.047)和抗组蛋白抗体(P=0.001)阳性率较F等位基因阴性SLE患者(ff基因型患者)明显升高。VDR mRNA在48例SLE患者表达下调,其?Ct值(?Ct值越大,表达量越小)为9.26±2.37,高于健康对照组的7.82±3.05(P=0.026)。而在SLE患者,携带F等位基因患者的VDR mRNA的?Ct值明显高于F等位基因阴性患者(10.54±1.88vs7.15±3.78,P=0.019)。结论:VDRFokⅠ多态性与SLE发病易感性有关,而且携带F等位基因的患者更容易发生浆膜炎和产生抗ds-DNA抗体和抗Sm抗体等自身抗体,此可能与F等位基因下调SLE患者的VDR mRNA表达有关。     

维生素D受体基因多态性与维生素D缺乏性佝偻病遗传关联性Meta分析

目的评价维生素D受体（VDR）基因多态性与维生素D缺乏性佝偻病（佝偻病）的遗传关联性。方法制定原始文献的纳入标准及检索策略,检索PubMed、Springer、Science Direct、Web of Science、中国期刊全文数据库、维普中文科技期刊数据库和万方数据库,收集VDR基因FokⅠ、ApaⅠ、BsmⅠ和TaqⅠ位点多态性与佝偻病相关性的病例对照研究,以佝偻病患儿为病例组。依据NHI-NHGRI研究工作组2007年制定的遗传关联性研究报告规范为基础,并依据相关文献选取其中的14条标准用于评价文献偏倚。以基因型频率为指标,提取数据后先确定最佳遗传模型,采用Stata 11.0软件进行Meta分析,计算合并的OR值及其95%CI。结果 19篇病例对照研究进入Meta分析。①FokⅠ位点采用共显性模型（FF基因型vsff基因型;FF基因型vsFf基因型）分析,病例组704例,对照组596例。Meta分析结果显示,亚洲人群FF基因型较ff基因型（OR=4.59,95%CI：2.98~7.07）和Ff基因型（OR=2.58,95%CI：1.79~3.73）患佝偻病的风险显著增加;高加索人群FokⅠ位点与佝偻病无显著关联性（FF基因型vsff基因型,OR=2.50,95%CI：0.76~8.19;FF基因型vsFf基因型,OR=1.18,95%CI：0.66~2.10）;非洲人群FF基因型较ff基因型患佝偻病的风险显著增加（OR=5.81,95%CI：1.21~27.98）。②ApaⅠ位点采用显性模型（AA＋Aa基因型vsaa基因型）分析,病例组338例,对照组459例。亚洲人群和非洲人群ApaⅠ位点与佝偻病均无显著关联性,OR分别为1.04（95%CI：0.72~1.49）和0.98（95%CI：0.57~1.71）;高加索人群AA＋Aa基因型患佝偻病的风险增高（OR=5.50,95%CI：1.22~24.75）。③BsmⅠ位点采用显性模型（bb基因型vsBb＋BB基因型）分析,病例组822例,对照组736例。亚洲人群BsmⅠ位点bb基因型较Bb＋BB基因型患佝偻病的风险降低（OR=0.46,95%CI：0.23~0.92）,非洲人群BsmⅠ位点与佝偻病无显著关联性（OR=1.65,95%CI：0.95~2.88）。④TaqⅠ位点采用隐性模型（TT基因型vsTt＋tt基因型）分析,病例组519例,对照组513例。亚洲人群（OR=1.22,95%CI：0.82~1.82）、高加索人群（OR=0.91,95%CI：0.35~2.35）和非洲人群（OR=1.18,95%CI：0.68~2.05）TaqⅠ位点与佝偻病无显著关联性。结论现有证据表明,亚洲人群FokⅠ位点FF基因型为患佝偻病的危险因素,而BsmⅠ位点bb基因型为佝偻病轻微的保护因素,尚不能认为ApaⅠ和TaqⅠ位点与佝偻病有关联性;由于高加索人群和非洲人群研究较少,VDR基因多态性与佝偻病的关联性尚不明确。     

阿昌族与汉族维生素D受体基因Fok多态性

目的　了解维生素 D受体基因多态性在中国不同民族中的分布。方法　应用聚合酶链反应 -限制性片段长度多态性分析、基因测序等技术检测 6 8名阿昌族人和 92名汉族人的维生素 D受体基因Fok 多态性 ,比较两组维生素 D受体基因型和等位基因的分布频率。结果　在 6 8名阿昌族人中 FF基因型占 18%、Ff基因型占 35 %、ff基因型占 4 7% ,而在 92名汉族人中 FF基因型占 2 2 %、Ff基因型占 5 2 %、ff基因型占 2 6 %。两组维生素 D受体基因型的分布频率差异有显著性 (χ2 =7.716 ,P=0 .0 2 1)。结论　阿昌族与汉族维生素 D受体基因 Fok 多态性分布频率差异有显著性。     

蒙古族人群维生素D受体基因Fok Ⅰ多态性

目的了解维生素D受体基因Fok Ⅰ多态性在蒙古族人群中的分布。方法应用聚合酶链反应一限制性片段长度多态性技术，分析506名内蒙古地区蒙古族人的维生素D受体Fok Ⅰ基因座的基因型和等位基因的分布频率。结果维生素D受体基因型FF、Ff、伍在本研究人群的分布频率分别为31％、52％和17％。等位基因F占57％，f占43％。基因型频率分布符合Hardy-Weinberg平衡定律。结论中国蒙古族人群维生素D受体基因Fok Ⅰ多态性分布频率有其自身的特点。     

蒙古族人群维生素D受体基因FokⅠ多态性

目的了解维生素D受体基因FokⅠ多态性在蒙古族人群中的分布。方法应用聚合酶链反应-限制性片段长度多态性技术,分析506名内蒙古地区蒙古族人的维生素D受体FokⅠ基因座的基因型和等位基因的分布频率。结果维生素D受体基因型FF、Ff、ff在本研究人群的分布频率分别为31%、52%和17%。等位基因F占57%,f占43%。基因型频率分布符合Hardy-Weinberg平衡定律。结论中国蒙古族人群维生素D受体基因FokⅠ多态性分布频率有其自身的特点。     

维生素D受体基因多态性与乙型肝炎病毒感染的关联研究

目的探讨中国汉族人群维生素D受体（vitamin D receptor，VDR）基因多态性是否与乙型肝炎病毒（hepatitis B virus，HBV）感染结局相关联。方法以184例慢性乙肝患者和205名无症状HBV携带者为研究对象，收集外周血，提取基因组DNA，应用聚合酶链反应．限制性片段长度多态性（polymerase chain reaction-restriction fragmentlength polymorphism，PCIR-RFLP）方法，分析VDR基因第2外显子Fok Ⅰ位点、第9外显子Taq Ⅰ位点的多态性分布。结果单因素分析结果显示Fok Ⅰ位点FF基因型在慢性乙肝组的频率44．6％显著高于无症状HBV携带组的31．7％（P〈0．05）。经多因素非条件Logistic回归分析调整混杂作用后，结果仍然显示FF基因型在慢性乙肝组与无症状HBV携带组之间的差异存在统计学意义（OR=1．95，P〈0．05）。FokⅠ位点与TapⅠ位点组成的FT单倍型在慢性乙肝组的分布频率显著高于无症状HBV携带组（OR=1．45，P〈0．05），fT单倍型在慢性乙肝组的分布频率显著低于无症状HBV携带组（OR=0．72，P〈0．05）。结论维生素D受体基因多态性可能影响HBV感染的遗传易感性。     

Single Nucleotide Polymorphism Typing of Global Salmonella enterica Serovar Typhi Isolates by Use of a Hairpin Primer Real-Time PCR Assay

Salmonella enterica serovar Typhi is highly homogeneous. Single nucleotide polymorphisms (SNPs) have been shown to be valuable markers for molecular typing of S. enterica serovar Typhi. Here, we used a hairpin primer real-time PCR assay for SNP typing of S. enterica serovar Typhi isolates. Forty-two SNPs were selected from a comparison of 19 published S. enterica serovar Typhi genomes and sequences from other studies. The SNPs were used to type 71 global S. enterica serovar Typhi isolates and differentiated these S. enterica serovar Typhi isolates and the 19 genome sequenced strains into 25 SNP profiles. Phylogenetic analysis revealed that these SNP profiles were grouped into six major clusters. These clusters can be identified by using five SNPs, while the full differentiation of the 25 SNP profiles requires a minimum of 24 SNPs. This real-time PCR-based SNP typing method will be useful for global epidemiological analysis.Salmonella enterica serovar Typhi is highly homogeneous (10, 17, 18). The lack of genetic diversity is a major challenge to the development of suitable typing methods to differentiate S. enterica serovar Typhi isolates for both phylogenetic and epidemiological purposes. Single nucleotide polymorphisms (SNPs) are considered the most valuable markers, particularly for studying the evolutionary relationships of isolates of homogeneous pathogenic clones, such as Bacillus anthracis (16), Mycobacterium tuberculosis (4), and Yersinia pestis (1).SNPs have been used as markers for molecular typing of S. enterica serovar Typhi in a large study by Roumagnac et al. (17). A total of 88 SNPs, found from analysis of 200 gene fragments from 105 diverse S. enterica serovar Typhi isolates, could differentiate 481 global S. enterica serovar Typhi isolates into 85 haplotypes (SNP profiles) and five major clusters (17). However, despite the large number of SNPs used, each of the five clusters was supported by only a single SNP and there was little resolution of the relationships of the haplotypes within a cluster. Eighty of the SNPs have also been used to differentiate 140 Indonesian S. enterica serovar Typhi isolates into nine haplotypes (2).We have also shown that genome-wide SNPs are useful for molecular typing and determining the relationships of global S. enterica serovar Typhi isolates (14). Thirty-seven SNPs selected from a comparison of the genomes of S. enterica serovar Typhi strains CT18 (15) and Ty2 (3) were typed using restriction enzyme digestion to differentiate 73 global S. enterica serovar Typhi isolates into 23 SNP profiles and four distinct genetic groups. As the SNPs were selected by comparison of only two S. enterica serovar Typhi genomes, this resulted in a phylogenetic bias which revealed the full path of the last common ancestors connecting strains CT18 and Ty2 but only the node locations for the other SNP profiles (14).Advances in technology, such as high-throughput sequencing, allow SNPs to be discovered to obtain a full resolution of the phylogenetic relationships of isolates. A recent study by Holt et al. (8) utilized 454 and/or Solexa technologies to sequence 19 S. enterica serovar Typhi isolates selected from the five major clusters found by Roumagnac et al. (17). There were more than 1,700 SNPs found, and these gave a fully resolved phylogenetic tree of these isolates. These SNPs are invaluable resources for investigation of the evolutionary history of global S. enterica serovar Typhi isolates. This study aimed to select a better set of SNPs on the basis of the genome tree and the previous SNP studies by Roumagnac et al. (17) to differentiate and establish the phylogenetic relationships of global S. enterica serovar Typhi isolates, using real-time (R-T) PCR assays based on hairpin (HP) primers (6).     

实时荧光定量PCR在检测人ANP基因表达的应用

目的建立心房钠尿肽（ANP）基因mRNA表达水平的实时荧光定量PCR检测方法,并对该方法进行初步评价。方法以基因表达产物为模板,建立实时荧光定量PCR检测方法,对样本中的心房钠尿肽含量进行相对定量,比较不同样本组的基因表达水平。结果所建立的实时荧光定量PCR方法熔解曲线中熔解峰单一。肺癌患者胸腔样本的ANP含量为对照样本的4.68倍,血清样本为对照样本的16.03倍。结论所建立的ANP实时荧光定量PCR检测方法具有较高的特异性。肺癌患者胸腔液和血清中ANP的含量明显增高。     

Serum 25-Hydroxy Vitamin D Levels and Association of Vitamin D Receptor Gene Polymorphisms in Vitiligo

BackgroundThe role of vitamin D deficiency and vitamin D receptor (VDR) gene polymorphisms has been established in many autoimmune diseases, including vitiligo, but the result is still controversial.ObjectivesThe aim of this study was to investigate the serum vitamin D levels in vitiligo patients and to compare the association of VDR gene polymorphisms in vitiligo patients and healthy controls.MethodsWe collected the data of age, sex, serum 25-hydroxy vitamin D (25[OH]D) level, thyroid autoantibodies, disease duration, types of vitiligo, family history and the affected body surface area of vitiligo from 172 patients. And we analyzed the VDR gene polymorphisms in 130 vitiligo and 453 age-sex-matched control subjects.ResultsThe mean serum level of 25(OH)D in 172 vitiligo patients was 18.75 ± 0.60 ng/mL, which had no significant difference with a mean serum value of 25(OH)D in the Korean population. However, there were significant differences according to the duration of the disease and family history. Also, there were no significant differences in the genotypic and allelic distributions of 37 examined SNPs of VDR gene between vitiligo patients and healthy controls.ConclusionSerum level of 25(OH)D in vitiligo patients was not significantly different from the mean serum value of the Korean population. Also, there were no significant differences in the genotypic distributions of VDR gene between vitiligo patients and healthy controls.     

Development of a New Molecular Subtyping Tool for Salmonella enterica Serovar Enteritidis Based on Single Nucleotide Polymorphism Genotyping Using PCR

The lack of a sufficiently discriminatory molecular subtyping tool for Salmonella enterica serovar Enteritidis has hindered source attribution efforts and impeded regulatory actions required to disrupt its food-borne transmission. The underlying biological reason for the ineffectiveness of current molecular subtyping tools such as pulsed-field gel electrophoresis (PFGE) and phage typing appears to be related to the high degree of clonality of S. Enteritidis. By interrogating the organism''s genome, we previously identified single nucleotide polymorphisms (SNP) distributed throughout the chromosome and have designed a highly discriminatory PCR-based SNP typing test based on 60 polymorphic loci. The application of the SNP-PCR method to DNA samples from S. Enteritidis strains (n = 55) obtained from a variety of sources has led to the differentiation and clustering of the S. Enteritidis isolates into 12 clades made up of 2 to 9 isolates per clade. Significantly, the SNP-PCR assay was able to further differentiate predominant PFGE types (e.g., XAI.0003) and phage types (e.g., phage type 8) into smaller subsets. The SNP-PCR subtyping test proved to be an accurate, precise, and quantitative tool for evaluating the relationships among the S. Enteritidis isolates tested in this study and should prove useful for clustering related S. Enteritidis isolates involved in outbreaks.     

Single Nucleotide Polymorphism in the Interleukin 12B Gene is Associated with Risk for Breast Cancer Development

Yellow nail syndrome (YNS) is a rare disorder of unknown aetiology that is characterized by yellow nails associated with lymphoedema and chronic respiratory manifestations. There are no detailed immunological studies in YNS. In this study, we present first extensive immunological analysis of both adaptive and innate immunity in two patients with YNS. One patient has common variable immunodeficiency, whereas second patient has specific antibody deficiency syndrome. Severe lymphopaenia, a striking deficiency of naïve CD4⁺ and CD8⁺ T cells and total B cells, and increased transitional B cells were observed. T cell proliferative response to mitogens and antigens was significantly reduced in both patients. Both patients failed to make specific antibody response to pneumococci. Complement, natural killer cell activity and neutrophil oxidative burst were normal. Immunoglobulin administration resulted in decreased frequency and severity of infections, and an impressive effect was observed on lymphoedema and on the recurrence of pleural effusion. Our data show that YNS is associated with both T and B cell defects. Furthermore, Immunoglobulin may be beneficial in clinical manifestations of lymphoedema.     

A Single Nucleotide Polymorphism of IL-21 Gene is Associated with Systemic Lupus Erythematosus in a Chinese Population

The aim of this study was to examine the association of single-nucleotide polymorphisms (SNPs) in IL-21 gene with susceptibility to systemic lupus erythematosus (SLE) in a Chinese population. A total of 605 independent SLE patients and 666 unrelated healthy controls were recruited for the case?Ccontrol association study. Two SNPs (rs2221903 and rs907715) within the IL-21 gene intronic region were genotyped by TaqMan SNP allelic discrimination methods. The allele T frequency of SNP rs2221903 in patients and healthy controls was 89.4?% and 86.8?%, respectively [T versus C, odds ratio (OR)?=?1.287, 95?% confidence interval (CI)?=?1.010?C1.640]. No significant differences in genotype frequencies were shown between SLE patients and healthy controls (P value?=?0.705, 0.406, respectively). However, the effect of recessive model (TT versus CC?+?CT, OR?=?1.368, 95?% CI?=?1.050?C1.781) was observed. Distributions of allele and genotype frequencies of the SNP rs907715 showed no significant differences between SLE patients and controls. Analysis of the haplotypes revealed that CC haplotype was significantly associated with SLE (OR?=?0.734, 95?% CI?=?0.573?C0.941). In conclusion, our findings suggest that a SNP (rs2221903) and CC haplotype (rs2221903 and rs907715) of the IL-21 gene is associated with SLE in the Chinese population. However, further studies are needed to determine the functional consequences of this polymorphism with SLE susceptibility.     

E-选择素基因第2外显子G98T单核苷酸多态性的调查

目的 :研究湖北地区汉族人群E 选择素 (E selectin)基因第 2外显子 98位点的单核苷酸多态性 (SNP) ,比较种族间单核苷酸的基因频率分布差异。方法 :应用聚合酶链反应 限制性片段长度多态性(PCR RFLP)的分析方法 ,检测了 2 40名健康者E selectin第 2外显子 98位点单核苷酸的基因型。结果 :E selectin各基因型频率GG型91.3 % ,GT型 8.7% ;G ,T各等位基因频率分别为 95 .6% ,4.4% ,这种基因多态性分布在男女间均无显著性差异 (P >0 .0 5 )。与其它种族比较 ,发现不同种族间E selectin基因型分布及等位基因频率均存在显著差异 (P <0 .0 5 )。结论 :在湖北地区汉族人群中存在E se lectin基因第 2外显子 98位点的单核苷酸多态性 ,这种多态性在种族间可能存在着较大的差异     

维生素D受体基因多态性与骨代谢疾病研究进展

维生素D受体(VDR)与骨代谢密切相关,在维持机体骨钙平衡中起着重要作用。对VDR基因型与骨钙代谢相关性研究,有助于推断骨质疏松、佝偻病等骨代谢疾病和遗传因素之间的关系,并且为从基因水平认识疾病的发病机理提供重要线索。     

Multiple Nucleotide Substitutions in the Neisseria meningitidis Serogroup C ctrA Gene Cause False-Negative Detection by Real-Time PCR

Two strains of Neisseria meningitidis serogroup C with disparate sequences of ctrA were isolated. The nucleotide substitutions did not alter the corresponding protein sequences, but they impeded the detection of these meningococcal isolates by real-time PCR.