AIM:To investigate the clinical implications of hepatitis B virus(HBV) pre S1 deletion.METHODS:We developed a fluorescence resonance energy transfer-based real-time polymerase chain reaction(RT-PCR) that can detect fo...AIM:To investigate the clinical implications of hepatitis B virus(HBV) pre S1 deletion.METHODS:We developed a fluorescence resonance energy transfer-based real-time polymerase chain reaction(RT-PCR) that can detect four genotypes(wild type, 15-bp, 18-bp and 21-bp deletion).The PCR method was used in two cohorts of Korean chronic HBV subjects with genotype C infections.Cohort Ⅰ included 292 chronic HBV subjects randomly selected from Cheju National University Hospital(Jeju, South Korea) or Seoul National University Hospital(Seoul, South Korea), and cohort Ⅱ included 90 consecutive chronic HBV carriers recruited from Konkuk University Hospital(Seoul, South Korea); the cohort Ⅱ patients did not have hepatocellular carcinoma or liver cirrhosis.RESULTS:The method proposed in this study identified 341 of 382 samples(89.3%).Deletion variants were identified in 100(29.3%) of the 341 detected samples.In both cohorts, the subjects with deletions had a significantly higher Hepatitis B virus e antigen(HBe Ag)-positive seroprevalence [cohort Ⅰ, wild(51.0%) vs deletion(75.0%), P < 0.001; cohort Ⅱ, wild(69.2%) vs deletion(92.9%), P = 0.002] and higher HBV DNA levels [cohort Ⅰ, wild(797.7 pg/m L) vs deletion(1678.9 pg/m L), P = 0.013; cohort Ⅱ, wild(8.3 × 108 copies/m L) vs deletion(2.2 × 109 copies/m L), P = 0.049], compared to subjects with wild type HBV.CONCLUSION:HBV genotype C pre S1 deletion may affect disease progression in chronic HBV subjects through an extended duration of HBe Ag seropositive status and increased HBV replications.展开更多
The DNA fragments coding for preS2(120 146) and preS1(21 47) amplified by PCR were fused to both 5′ and 3′ ends of S gene at the position of amino acid 223. The fusion gene was placed downstream of the promoter P7.5...The DNA fragments coding for preS2(120 146) and preS1(21 47) amplified by PCR were fused to both 5′ and 3′ ends of S gene at the position of amino acid 223. The fusion gene was placed downstream of the promoter P7.5 of the universal vaccinia viral vector pGJP 5 and the recombinant vaccinia virus vS2SS1 was then selected by \%in vivo\% homogeneous recombination. Fusion protein S2SS1 could be expressed in the mammalian cells infected with vS2SS1. The investigation of expression, secretion, antigenicity and particle assembly of the S2SS1 protein demonstrated that S2SS1 protein could be assembled into particles which presented preS1, preS2 and S antigenicity and be efficiently secreted from the cells. It also showed that the level of its expression and secretion approached to that of the S protein expressed by the recombinant vaccinia virus.展开更多
A synthetic peptide, the N\|terminus of hepatitis B virus surface antigen Pre\|S1, was studied by two\| dimensional NMR techniques. A series of 1 H nuclear magnetic resonance experiments were used to complete the iden...A synthetic peptide, the N\|terminus of hepatitis B virus surface antigen Pre\|S1, was studied by two\| dimensional NMR techniques. A series of 1 H nuclear magnetic resonance experiments were used to complete the identification of spin systems and sequential assignments of this 28\|residue peptide. 157 distance constraints and 55 dihedral angle constraints were obtained. 20 structures with the lowest target function were selected by the distance geometry program DIANA. Energy minimization and the following 100 ps time\|averaged restrained molecular dynamics （TRMD） simulation in aqueous solution were performed for each conformer. After TRMD simulation, three locally convergent regions corresponding to residues 22\31, 36\40, 41\46 were found. The averaged pairwise root\|mean\|square deviation （RMSD） of backbone atoms for them were （1.71±0.49）?, （0.76±0.31）?, （1.05±0.52）?, respectively. Four reverse turns found in these regions, residues 22\25, 37\40, 41\44 and 43\46, correspond to several important antibody binding sites revealed in relevant immunological research.展开更多
目的研究蜕膜各淋巴细胞亚群中程序性细胞死亡蛋白1(programmed cell death protein 1,PD-1)、程序性细胞死亡蛋白配体1(programmed cell death ligand 1,PD-L1)和淋巴细胞活化基因3(lymphocyte-activation gene 3,LAG-3)的表达,及其与...目的研究蜕膜各淋巴细胞亚群中程序性细胞死亡蛋白1(programmed cell death protein 1,PD-1)、程序性细胞死亡蛋白配体1(programmed cell death ligand 1,PD-L1)和淋巴细胞活化基因3(lymphocyte-activation gene 3,LAG-3)的表达,及其与子痫前期免疫失衡的关系。方法纳入2019年5月至2020年1月于山东第一医科大学附属省立医院剖宫产分娩的单胎妊娠子痫前期患者25例为子痫前期组,按照1∶1的比例选取剖宫产日期和分娩孕周匹配的产前检查正常并择期剖宫产分娩的健康单胎妊娠孕妇25例为正常妊娠组。剖宫产胎盘娩出后收集蜕膜组织。采用流式细胞技术测定PD-1、PD-L1和LAG-3在蜕膜T细胞、自然杀伤(natural killer,NK)和自然杀伤T(natural killer T,NKT)细胞表面的表达水平。采用两独立样本t检验比较2组间PD-1、PD-L1和LAG-3的表达差异。结果子痫前期组蜕膜中PD-1在T细胞和NK细胞表面的表达水平低于正常妊娠组(37.84±3.82与57.02±3.89,t=3.529,P<0.001;3.28±0.48与5.69±0.99,t=2.184,P=0.034),而在NKT细胞表面的表达水平2组差异无统计学意义(P=0.461)。子痫前期组蜕膜中PD-L1在NK细胞表面的表达水平低于正常妊娠组(0.60±0.11与1.32±0.19,t=3.319,P=0.002),而在T细胞和NKT细胞表面的表达水平2组差异无统计学意义(P值均>0.05)。子痫前期组蜕膜中LAG-3在T细胞和NKT细胞表面的表达水平低于正常妊娠组(2.32±0.36与4.09±0.67,t=2.335,P=0.024;35.40±4.97与56.27±4.49,t=3.282,P=0.002),而在NK细胞表面的表达水平2组差异无统计学意义(P=0.112)。结论免疫检查点PD-1、PD-L1和LAG-3在子痫前期蜕膜淋巴细胞亚群中表达水平明显降低,可能通过母胎界面免疫细胞过度激活参与子痫前期的免疫失衡。展开更多
基金Supported by Grants from National Research Foundation of Koreagrant funded by the Korean government(Ministry of Education,Science,and Technology),No.2013-005810Foundation of Seoul National University Hospital(SNUH research fund),No.0320140140
文摘AIM:To investigate the clinical implications of hepatitis B virus(HBV) pre S1 deletion.METHODS:We developed a fluorescence resonance energy transfer-based real-time polymerase chain reaction(RT-PCR) that can detect four genotypes(wild type, 15-bp, 18-bp and 21-bp deletion).The PCR method was used in two cohorts of Korean chronic HBV subjects with genotype C infections.Cohort Ⅰ included 292 chronic HBV subjects randomly selected from Cheju National University Hospital(Jeju, South Korea) or Seoul National University Hospital(Seoul, South Korea), and cohort Ⅱ included 90 consecutive chronic HBV carriers recruited from Konkuk University Hospital(Seoul, South Korea); the cohort Ⅱ patients did not have hepatocellular carcinoma or liver cirrhosis.RESULTS:The method proposed in this study identified 341 of 382 samples(89.3%).Deletion variants were identified in 100(29.3%) of the 341 detected samples.In both cohorts, the subjects with deletions had a significantly higher Hepatitis B virus e antigen(HBe Ag)-positive seroprevalence [cohort Ⅰ, wild(51.0%) vs deletion(75.0%), P < 0.001; cohort Ⅱ, wild(69.2%) vs deletion(92.9%), P = 0.002] and higher HBV DNA levels [cohort Ⅰ, wild(797.7 pg/m L) vs deletion(1678.9 pg/m L), P = 0.013; cohort Ⅱ, wild(8.3 × 108 copies/m L) vs deletion(2.2 × 109 copies/m L), P = 0.049], compared to subjects with wild type HBV.CONCLUSION:HBV genotype C pre S1 deletion may affect disease progression in chronic HBV subjects through an extended duration of HBe Ag seropositive status and increased HBV replications.
文摘The DNA fragments coding for preS2(120 146) and preS1(21 47) amplified by PCR were fused to both 5′ and 3′ ends of S gene at the position of amino acid 223. The fusion gene was placed downstream of the promoter P7.5 of the universal vaccinia viral vector pGJP 5 and the recombinant vaccinia virus vS2SS1 was then selected by \%in vivo\% homogeneous recombination. Fusion protein S2SS1 could be expressed in the mammalian cells infected with vS2SS1. The investigation of expression, secretion, antigenicity and particle assembly of the S2SS1 protein demonstrated that S2SS1 protein could be assembled into particles which presented preS1, preS2 and S antigenicity and be efficiently secreted from the cells. It also showed that the level of its expression and secretion approached to that of the S protein expressed by the recombinant vaccinia virus.
文摘A synthetic peptide, the N\|terminus of hepatitis B virus surface antigen Pre\|S1, was studied by two\| dimensional NMR techniques. A series of 1 H nuclear magnetic resonance experiments were used to complete the identification of spin systems and sequential assignments of this 28\|residue peptide. 157 distance constraints and 55 dihedral angle constraints were obtained. 20 structures with the lowest target function were selected by the distance geometry program DIANA. Energy minimization and the following 100 ps time\|averaged restrained molecular dynamics （TRMD） simulation in aqueous solution were performed for each conformer. After TRMD simulation, three locally convergent regions corresponding to residues 22\31, 36\40, 41\46 were found. The averaged pairwise root\|mean\|square deviation （RMSD） of backbone atoms for them were （1.71±0.49）?, （0.76±0.31）?, （1.05±0.52）?, respectively. Four reverse turns found in these regions, residues 22\25, 37\40, 41\44 and 43\46, correspond to several important antibody binding sites revealed in relevant immunological research.
文摘目的研究蜕膜各淋巴细胞亚群中程序性细胞死亡蛋白1(programmed cell death protein 1,PD-1)、程序性细胞死亡蛋白配体1(programmed cell death ligand 1,PD-L1)和淋巴细胞活化基因3(lymphocyte-activation gene 3,LAG-3)的表达,及其与子痫前期免疫失衡的关系。方法纳入2019年5月至2020年1月于山东第一医科大学附属省立医院剖宫产分娩的单胎妊娠子痫前期患者25例为子痫前期组,按照1∶1的比例选取剖宫产日期和分娩孕周匹配的产前检查正常并择期剖宫产分娩的健康单胎妊娠孕妇25例为正常妊娠组。剖宫产胎盘娩出后收集蜕膜组织。采用流式细胞技术测定PD-1、PD-L1和LAG-3在蜕膜T细胞、自然杀伤(natural killer,NK)和自然杀伤T(natural killer T,NKT)细胞表面的表达水平。采用两独立样本t检验比较2组间PD-1、PD-L1和LAG-3的表达差异。结果子痫前期组蜕膜中PD-1在T细胞和NK细胞表面的表达水平低于正常妊娠组(37.84±3.82与57.02±3.89,t=3.529,P<0.001;3.28±0.48与5.69±0.99,t=2.184,P=0.034),而在NKT细胞表面的表达水平2组差异无统计学意义(P=0.461)。子痫前期组蜕膜中PD-L1在NK细胞表面的表达水平低于正常妊娠组(0.60±0.11与1.32±0.19,t=3.319,P=0.002),而在T细胞和NKT细胞表面的表达水平2组差异无统计学意义(P值均>0.05)。子痫前期组蜕膜中LAG-3在T细胞和NKT细胞表面的表达水平低于正常妊娠组(2.32±0.36与4.09±0.67,t=2.335,P=0.024;35.40±4.97与56.27±4.49,t=3.282,P=0.002),而在NK细胞表面的表达水平2组差异无统计学意义(P=0.112)。结论免疫检查点PD-1、PD-L1和LAG-3在子痫前期蜕膜淋巴细胞亚群中表达水平明显降低,可能通过母胎界面免疫细胞过度激活参与子痫前期的免疫失衡。
