Objective:To investigate the influence of ABCB1 polymorphisms on the plasma level of efavirenz in Thai adult cases infected with HIV-1.Methods:A single nucleotide polymorphism of ABCB13435 C>T(rs1045642)in the gene...Objective:To investigate the influence of ABCB1 polymorphisms on the plasma level of efavirenz in Thai adult cases infected with HIV-1.Methods:A single nucleotide polymorphism of ABCB13435 C>T(rs1045642)in the gene encoding ABCB1 was genotyped using real-time PCR-based alleles in 149 HIV-infected Thai adults receiving efavirenz treatment.Plasma concentrations of efavirenz were measured by high-performance liquid chromatography 12 hr after administration.The relationship between plasma efavirenz concentrations and ABCB13435 C>T polymorphisms was analyzed.Results:Logistic regression analysis showed no significant predictors of high plasma efavirenz concentration in relation to age,gender,body weight,CD4 count and plasma HIV-1 RNA,blood biochemical parameters,antiretroviral duration or ABCB13435 C>T polymorphisms,except for height(OR=0.902,95%CI:0.835-0.973)(P<0.05).The minor allele frequency of ABCB13435 C>T was0.446.The frequency of the heterozygous mutant ABCB13435 C/T was 53.02%(n=79),ABCB13435 T/T homozygous mutant was 18.12%(n=21)and the wild type ABCB13435 C/C genotype was 28.86%(n=43).The overall median plasma concentration of efavirenz in 149 HIV-infected Thai cases was 2.41 mg/L[IQR:(1.46-4.12)mg/L].The plasma concentration of efavirenz was higher in cases with ABCB13435 T/T homozygous mutant[2.73 mg/L,IQR:(2.02-4.19)mg/L]and ABCB13435 C/T heterozygous mutant[2.29 mg/L,IQR:(1.41-4.28)mg/L]genotypes compared to the wild type ABCB13435 C/C homozygous[2.1 mg/L,IQR:(1.37-3.53)mg/L].However,there was no statistically significant difference in the efavirenz concentration between the different genotypes(P>0.05).Objective:To investigate the influence of ABCB1 polymorphisms on the plasma level of efavirenz in Thai adult cases infected with HIV-1.Methods:A single nucleotide polymorphism of ABCB13435 C>T(rs1045642)in the gene encoding ABCB1 was genotyped using real-time PCR-based alleles in 149 HIV-infected Thai adults receiving efavirenz treatment.Plasma concentrations of efavirenz were measured by high-performance liquid chromatography 12 hr after administration.The relationship between plasma efavirenz concentrations and ABCB13435 C>T polymorphisms was analyzed.Results:Logistic regression analysis showed no significant predictors of high plasma efavirenz concentration in relation to age,gender,body weight,CD4 count and plasma HIV-1 RNA,blood biochemical parameters,antiretroviral duration or ABCB13435 C>T polymorphisms,except for height(OR=0.902,95%CI:0.835-0.973)(P<0.05).The minor allele frequency of ABCB13435 C>T was 0.446.The frequency of the heterozygous mutant ABCB13435 C/T was 53.02%(n=79),ABCB13435 T/T homozygous mutant was 18.12%(n=27)and the wild type ABCB13435 C/C genotype was 28.86%(n=43).The overall median plasma concentration of efavirenz in 149 HIV-infected Thai cases was 2.41 mg/L[IQR:(1.46-4.12)mg/L].The plasma concentration of efavirenz was higher in cases with ABCB13435 T/T homozygous mutant[2.73 mg/L,IQR:(2.02-4.19)mg/L]and ABCB13435 C/T heterozygous mutant[2.29 mg/L,IQR:(1.41-4.28)mg/L]genotypes compared to the wild type ABCB13435 C/C homozygous[2.1 mg/L,IQR:(1.37-3.53)mg/L].However,there was no statistically significant difference in the efavirenz concentration between the different genotypes(P>0.05).Conclusions:There is no statistical significance for a tendency toward higher plasma efavirenz concentration in the ABCB13435 T/T and ABCB13435 C/T genotypes.No parameters of physiological characteristics in this study except for height were found to be predictors of high plasma efavirenz concentration in Thai HIV-1 infected cases.展开更多
In order to isolate expeditiously the HIV1U26942 DNA fragments for preparation of DNA microarrays, the multiple gene fragments with sizes suitable for DNA microarrays, produced by digesting the dissociated HIV gene wi...In order to isolate expeditiously the HIV1U26942 DNA fragments for preparation of DNA microarrays, the multiple gene fragments with sizes suitable for DNA microarrays, produced by digesting the dissociated HIV gene with Sau 3AⅠ, were ligated with universal adapters. PCR primers were designed to match the universal adapters (including the restriction site sequence) but with one "nesting" base overhanging at the 3’ end. The PCR reactions that were performed with various single primers or primer combinations were divided into ten subgroups. PCR products were purified and then cloned into the T vectors. The positive clones were propagated and the plasmids were extracted. The target HIV gene fragments were isolated and sequenced, which were correlated precisely with the prediction of restriction analysis. Eighteen gene fragments ranging from 0.1kb to 1kb were prepared for DNA microarray. Restriction display is an effective and rapid method for the isolation of gene展开更多
目的对保定市新发现的1株pol区不能明确分型的HIV-1毒株(BD226AJ)进行近似全长基因组扩增,并分析其亚型、重组模式和基因特点。方法提取患者血浆中HIV-1RNA并逆转录为cDNA,使用近末端稀释法分2段对其进行近似全长基因组扩增并测序。使用...目的对保定市新发现的1株pol区不能明确分型的HIV-1毒株(BD226AJ)进行近似全长基因组扩增,并分析其亚型、重组模式和基因特点。方法提取患者血浆中HIV-1RNA并逆转录为cDNA,使用近末端稀释法分2段对其进行近似全长基因组扩增并测序。使用jpHMM和SimPlot 3.5软件对近似全长基因组序列进行重组模式和重组断点分析,采用MEGA 6.0软件分片段构建Neighbor-joining系统进化树进一步确认重组断点的准确性。构建该近似全长基因组序列及各亚型片段Neighbor-joining系统进化树,分析该毒株的亲本来源。结果经过近似全长基因组扩增、测序、序列拼接后,获得1条长度为8830 bp的HIV-1近似全长基因组序列。重组分析结果显示,该序列是由CRF01_AE和B亚型重组形成的,其近似全长基因组序列被3个断点分成了4个亚型片段,分别为ICRF01_AE(HXB2,823—4224 nt)、Ⅱ_(B)(HXB2,4225—5991 nt)、ⅢCRF01_AE(HX B2,5992—9295 nt)、ⅣB(HXB2,9296—9406 nt)。各亚型基因片段的系统进化树分析进一步表明该序列的可能亲本来源为CRF01_AE和B亚型。HIV BLAST的结果显示,该序列与CRF112_01B的相似性为96%,系统进化树分析进一步验证该序列与北京市男男性行为者(men who have sex with men,MSM)中的CRF112_01B序列聚集。结论本研究在保定市MSM人群中发现了1例由CRF01_AE和B亚型重组的新型重组毒株CRF112_01B,提示HIV-1CRF112_01B已通过MSM人群传入河北,并开始在保定市传播,因此加强该亚型或者类似新型毒株的监测,对有关部门采取针对性防控措施和遏制新型重组毒株在本地区的传播和流行具有重要意义。展开更多
人类免疫缺陷病毒1型(Human immunodeficiency virus type 1,HIV-1)是人获得性免疫缺陷综合征即艾滋病(AIDS)的病原,HIV-1感染人类免疫细胞,渐进性地破坏免疫系统,最终引发艾滋病。ANP32A (酸性核磷蛋白32A)与ANP32B同属于ANP32蛋白家族...人类免疫缺陷病毒1型(Human immunodeficiency virus type 1,HIV-1)是人获得性免疫缺陷综合征即艾滋病(AIDS)的病原,HIV-1感染人类免疫细胞,渐进性地破坏免疫系统,最终引发艾滋病。ANP32A (酸性核磷蛋白32A)与ANP32B同属于ANP32蛋白家族,是近几年新发现的与流感病毒复制相关的宿主蛋白。前期有研究证明宿主因子ANP32A/B是A型流感病毒聚合酶发挥功能所必需的宿主因子,并揭示了其与聚合酶相互作用的关键位点,为新型抗流感药物及转基因动物的研发提供了有效靶点(Zhang H et al. Journal of Virology 2019)。展开更多
基金supported by Mahidol University,Thailand Research Fund,Thailand Office of the Higher Education Commission under New Researchers Grant(MRG 5480136)the project CICECOAveiro Institute of Materials,national funds through the FCT/MCTES(FCT Ref.UID/CTM/50011/2019)Rachadapisek Sompote Fund for Postdoctoral Fellowship,Chulalongkorn University
文摘Objective:To investigate the influence of ABCB1 polymorphisms on the plasma level of efavirenz in Thai adult cases infected with HIV-1.Methods:A single nucleotide polymorphism of ABCB13435 C>T(rs1045642)in the gene encoding ABCB1 was genotyped using real-time PCR-based alleles in 149 HIV-infected Thai adults receiving efavirenz treatment.Plasma concentrations of efavirenz were measured by high-performance liquid chromatography 12 hr after administration.The relationship between plasma efavirenz concentrations and ABCB13435 C>T polymorphisms was analyzed.Results:Logistic regression analysis showed no significant predictors of high plasma efavirenz concentration in relation to age,gender,body weight,CD4 count and plasma HIV-1 RNA,blood biochemical parameters,antiretroviral duration or ABCB13435 C>T polymorphisms,except for height(OR=0.902,95%CI:0.835-0.973)(P<0.05).The minor allele frequency of ABCB13435 C>T was0.446.The frequency of the heterozygous mutant ABCB13435 C/T was 53.02%(n=79),ABCB13435 T/T homozygous mutant was 18.12%(n=21)and the wild type ABCB13435 C/C genotype was 28.86%(n=43).The overall median plasma concentration of efavirenz in 149 HIV-infected Thai cases was 2.41 mg/L[IQR:(1.46-4.12)mg/L].The plasma concentration of efavirenz was higher in cases with ABCB13435 T/T homozygous mutant[2.73 mg/L,IQR:(2.02-4.19)mg/L]and ABCB13435 C/T heterozygous mutant[2.29 mg/L,IQR:(1.41-4.28)mg/L]genotypes compared to the wild type ABCB13435 C/C homozygous[2.1 mg/L,IQR:(1.37-3.53)mg/L].However,there was no statistically significant difference in the efavirenz concentration between the different genotypes(P>0.05).Objective:To investigate the influence of ABCB1 polymorphisms on the plasma level of efavirenz in Thai adult cases infected with HIV-1.Methods:A single nucleotide polymorphism of ABCB13435 C>T(rs1045642)in the gene encoding ABCB1 was genotyped using real-time PCR-based alleles in 149 HIV-infected Thai adults receiving efavirenz treatment.Plasma concentrations of efavirenz were measured by high-performance liquid chromatography 12 hr after administration.The relationship between plasma efavirenz concentrations and ABCB13435 C>T polymorphisms was analyzed.Results:Logistic regression analysis showed no significant predictors of high plasma efavirenz concentration in relation to age,gender,body weight,CD4 count and plasma HIV-1 RNA,blood biochemical parameters,antiretroviral duration or ABCB13435 C>T polymorphisms,except for height(OR=0.902,95%CI:0.835-0.973)(P<0.05).The minor allele frequency of ABCB13435 C>T was 0.446.The frequency of the heterozygous mutant ABCB13435 C/T was 53.02%(n=79),ABCB13435 T/T homozygous mutant was 18.12%(n=27)and the wild type ABCB13435 C/C genotype was 28.86%(n=43).The overall median plasma concentration of efavirenz in 149 HIV-infected Thai cases was 2.41 mg/L[IQR:(1.46-4.12)mg/L].The plasma concentration of efavirenz was higher in cases with ABCB13435 T/T homozygous mutant[2.73 mg/L,IQR:(2.02-4.19)mg/L]and ABCB13435 C/T heterozygous mutant[2.29 mg/L,IQR:(1.41-4.28)mg/L]genotypes compared to the wild type ABCB13435 C/C homozygous[2.1 mg/L,IQR:(1.37-3.53)mg/L].However,there was no statistically significant difference in the efavirenz concentration between the different genotypes(P>0.05).Conclusions:There is no statistical significance for a tendency toward higher plasma efavirenz concentration in the ABCB13435 T/T and ABCB13435 C/T genotypes.No parameters of physiological characteristics in this study except for height were found to be predictors of high plasma efavirenz concentration in Thai HIV-1 infected cases.
文摘In order to isolate expeditiously the HIV1U26942 DNA fragments for preparation of DNA microarrays, the multiple gene fragments with sizes suitable for DNA microarrays, produced by digesting the dissociated HIV gene with Sau 3AⅠ, were ligated with universal adapters. PCR primers were designed to match the universal adapters (including the restriction site sequence) but with one "nesting" base overhanging at the 3’ end. The PCR reactions that were performed with various single primers or primer combinations were divided into ten subgroups. PCR products were purified and then cloned into the T vectors. The positive clones were propagated and the plasmids were extracted. The target HIV gene fragments were isolated and sequenced, which were correlated precisely with the prediction of restriction analysis. Eighteen gene fragments ranging from 0.1kb to 1kb were prepared for DNA microarray. Restriction display is an effective and rapid method for the isolation of gene
文摘目的对保定市新发现的1株pol区不能明确分型的HIV-1毒株(BD226AJ)进行近似全长基因组扩增,并分析其亚型、重组模式和基因特点。方法提取患者血浆中HIV-1RNA并逆转录为cDNA,使用近末端稀释法分2段对其进行近似全长基因组扩增并测序。使用jpHMM和SimPlot 3.5软件对近似全长基因组序列进行重组模式和重组断点分析,采用MEGA 6.0软件分片段构建Neighbor-joining系统进化树进一步确认重组断点的准确性。构建该近似全长基因组序列及各亚型片段Neighbor-joining系统进化树,分析该毒株的亲本来源。结果经过近似全长基因组扩增、测序、序列拼接后,获得1条长度为8830 bp的HIV-1近似全长基因组序列。重组分析结果显示,该序列是由CRF01_AE和B亚型重组形成的,其近似全长基因组序列被3个断点分成了4个亚型片段,分别为ICRF01_AE(HXB2,823—4224 nt)、Ⅱ_(B)(HXB2,4225—5991 nt)、ⅢCRF01_AE(HX B2,5992—9295 nt)、ⅣB(HXB2,9296—9406 nt)。各亚型基因片段的系统进化树分析进一步表明该序列的可能亲本来源为CRF01_AE和B亚型。HIV BLAST的结果显示,该序列与CRF112_01B的相似性为96%,系统进化树分析进一步验证该序列与北京市男男性行为者(men who have sex with men,MSM)中的CRF112_01B序列聚集。结论本研究在保定市MSM人群中发现了1例由CRF01_AE和B亚型重组的新型重组毒株CRF112_01B,提示HIV-1CRF112_01B已通过MSM人群传入河北,并开始在保定市传播,因此加强该亚型或者类似新型毒株的监测,对有关部门采取针对性防控措施和遏制新型重组毒株在本地区的传播和流行具有重要意义。
文摘人类免疫缺陷病毒1型(Human immunodeficiency virus type 1,HIV-1)是人获得性免疫缺陷综合征即艾滋病(AIDS)的病原,HIV-1感染人类免疫细胞,渐进性地破坏免疫系统,最终引发艾滋病。ANP32A (酸性核磷蛋白32A)与ANP32B同属于ANP32蛋白家族,是近几年新发现的与流感病毒复制相关的宿主蛋白。前期有研究证明宿主因子ANP32A/B是A型流感病毒聚合酶发挥功能所必需的宿主因子,并揭示了其与聚合酶相互作用的关键位点,为新型抗流感药物及转基因动物的研发提供了有效靶点(Zhang H et al. Journal of Virology 2019)。
