Background The present study was undertaken to replicate the associations of representative polymorphisms in three genes （complement factor H （CFH）, complement factor B （BF） and HtrA serine peptidase 1 （HTRA1）...Background The present study was undertaken to replicate the associations of representative polymorphisms in three genes （complement factor H （CFH）, complement factor B （BF） and HtrA serine peptidase 1 （HTRA1）） with exudative age-related macular degeneration （AMD） in a Han Chinese population, and to test if the modifiable environmental factors affect AMD susceptibility associated with different type of genotype in these genes. Methods An age, gender and ethnicity matched case-control study was conducted to genotype the representative single neucleotide polymorphisms （SNPs） loci including rs1061170 and rs1410996 in CFH, rs641153 and rs4151667 in BF and rs11200638 in HTRA1 gene in 144 exudative AMD patients and 126 normal controls using PCR-RFLP and direct resequencing. The demographic characteristics and behavioral risk factors were also recorded. Allelic and genotypic associations for individual SNP and joint associations with two loci were performed. The gene-gene and gene-environment interactions were analyzed using multivariate non-conditional Logistic regression analysis. Results The C risk allele frequencies for CFH Y402H （rs1061170） in cases and controls were 12.5% and 5.4% respectively, which were much lower than those in Caucasians （P 〈0.001）. Compared with TT homozygous genotype, the CT heterozygous genotype was positively associated with AMD with odds ratio （OR） of 3.23 （1.36-5.07）. However, the population attributable risk （PAR） of C allele was only 3.3% （1.4%-4.3%）. rs1410996 was also associated with AMD independent of Y402H. The ORs of exudative AMD for individuals carrying one copy risk allele and two copy risk alleles were 2.57 （1.21-5.45） and 4.76 （2.15-10.55） respectively, with correspondent PARs of 28.3% （2.0%-40.5%） and 38.2% （21.8%-45.4%）. rs11200636 in HTRA1 was another susceptible locus for AMD and the risk homozygotes were significantly susceptible for exudutive AMD （OR=3.98, 1.88-8.43） with PAR of 38.9% （24.3%-45.8%）展开更多
子痫前期(preeclampsia)是妊娠期特有的疾病,目前仍是重要的公共卫生问题,是发展中国家孕产妇死亡的主要原因之一。研究表明子痫前期与细胞凋亡、血管痉挛、凝血系统激活、炎症反应等密切相关,其病理变化起始于胎盘浅着床和胎盘血管重...子痫前期(preeclampsia)是妊娠期特有的疾病,目前仍是重要的公共卫生问题,是发展中国家孕产妇死亡的主要原因之一。研究表明子痫前期与细胞凋亡、血管痉挛、凝血系统激活、炎症反应等密切相关,其病理变化起始于胎盘浅着床和胎盘血管重铸障碍,导致胎盘组织的缺血缺氧,引起子痫前期的发生。HtrA(high temperature requirement A)系丝氨酸蛋白酶家族,该蛋白家族在细胞热休克、氧化应激、炎症、缺血再灌注和肿瘤等应激条件下,降解细胞胞质内错误折叠的蛋白质,参与细胞凋亡、关节炎、胚胎形成、神经变性和神经肌肉障碍以及癌症等的病理生理过程,研究表明其亚型与子痫前期密切相关。总结近年来与子痫前期有关的HtrA的研究进展及其介导的调控网络,对进一步了解子痫前期的发病机制及提供新的治疗途径有重要意义。展开更多
To investigate the roles of HtrA protein in bacterial adaption to stresses, htra gene was cloned from an Escherichia coli strain isolated from silver fox, double digested, and ligated into the prokaryotic expression v...To investigate the roles of HtrA protein in bacterial adaption to stresses, htra gene was cloned from an Escherichia coli strain isolated from silver fox, double digested, and ligated into the prokaryotic expression vector pET32a. Then, the recombinant plasmid pET32a-htra was transformed into E. coli BL21 competent cells. The over-expression of His-HtrA protein was induced by the addition of IPTG at a final concentration of 1 mmol/L, and verified by SDA-PAGE and Western blot. In addition, the protein was purified with Ni-NTA agarose, and used to immunize mice,and the polyclonal antibody specifically bound to E. coli HtrA protein. The results will provide a theoretical basis for studying the roles of HtrA protein and developing vaccines for fur-bearing animals against E. coli.展开更多
Objective:To explore HtrA1 gene expression aud its regulation in human gastric cancers.Methods:The HtrA1 mRNA levels were examined by QPCR analysis and coufirmed its expression with Northern blot analysis.The HtrA1 pr...Objective:To explore HtrA1 gene expression aud its regulation in human gastric cancers.Methods:The HtrA1 mRNA levels were examined by QPCR analysis and coufirmed its expression with Northern blot analysis.The HtrA1 protein levels in all six gastric epithelial cell lines were investigated by Western blot analysis.Gene copy number was accessed and then sequenced the coding region from each mRNA in all six cell lines.The HtrA1 promoter region DNA methylation status was detected by using bisulfite sequeucing analysis.Effect of decitabine and TSA on HTRA1 expression in gastric cancer cell line was determined by RTPCR.Results:HIC analysis indicated that HtrA1 was highly expressed in normal epithelium,but dramatically down-regulated in gastric carcinoma tissues and variably expressed in tumor-adjacent tissues.HtrA1 gene expression was dramatically decreased in gastric carcinoma cells compared to nontumorigenic counterparts.The HtrA1 gene loss in any of the 4 breast cancer cell lines was not detected.Total 14 CpGs in this region were all methylated in gastric cancer cells,whereas two normal cells.GES-1 and HFI-145,were having several unmethylated cytosines in this region.HtrA1 showed as^Mr 44,000,Expression of HtrA1 protein was not observed in any of the four gastric caucer cell lines.BGC-823.MKN-45.SGC-7901and MKN-28.HtrA1 expression was observed in the HF1-145and GES-1 cell lines.Conclusions:The epigenetic silencing for HtrA1gene expression could provide a possible strategy for re-activating Htrt1 gene expression in gastric cancer cells.thus facilitating further investigation of HtrA1's role in chemotherapy.展开更多
Legionella pneumophila,the causative agent of Legionnaires' disease,has been recognized as a major health problem responsible for an estimated number of 15 000-30 000 cases of severe pneumonia per year in Germany ...Legionella pneumophila,the causative agent of Legionnaires' disease,has been recognized as a major health problem responsible for an estimated number of 15 000-30 000 cases of severe pneumonia per year in Germany alone.Despite of the high clinical relevance,many aspects of the intracellular life cycle of Legionella,especially details on interactions with host cells,are not well understood.Structural information on virulence proteins helps unravel basal pathogenicity mechanisms and is a prerequisite for the rational development of effective drug molecules.Here we discuss structures of three important virulence proteins of Legionella that have been determined in our laboratory.The structure of the macrophage infectivity potentiator(Mip) protein of Legionella pneumophila is the first of a novel subgroup within the family of FK506-binding protein(FKBP) peptidyl-prolyl cis/trans isomerases.On the basis of the Mip structure,promising antibacterial agents are being designed.Recently,structures of two equally exciting Legionella proteins have been reported.The ferrous iron transport protein FeoB is a transmembrane protein responsible for Fe2+ aquisition after entry of the pathogen into the host cell.The structure of the cytoplasmic domain of FeoB provides insights into the family of prokaryotic G proteins and allows a detailed comparison with structures of related FeoBs.Furthermore,the characterization of DegQ,a periplasmatic chaperone-protease involved in protein quality control represents an intriguing example of how enzymatic activity is regulated by oligomerization as well as by an intrinsic loop activation cascade,depending on subtle conformational rearrangements.展开更多
目的:在体外非应激条件下,研究乳牙变异链球菌(streptococcus mutans,S.mutans)高温需要A蛋白(high temperature requirement serine proteinase A,HtrA)基因缺陷株和HtrA高毒力株的葡聚糖结合蛋白C(glucan-binding protein C,gbpC)表...目的:在体外非应激条件下,研究乳牙变异链球菌(streptococcus mutans,S.mutans)高温需要A蛋白(high temperature requirement serine proteinase A,HtrA)基因缺陷株和HtrA高毒力株的葡聚糖结合蛋白C(glucan-binding protein C,gbpC)表达能力的差异,探讨HtrA基因对gbpC表达的影响。方法:选用前期已获得的乳牙HtrA高毒力株和高致龋性HtrA基因缺陷株,在BHI培养基培养至指数期第12 h后进行细菌革兰染色及生化鉴定。以qPCR和Western Blot分别检测两组S.mutans菌株中gbpC基因和蛋白的表达情况。结果:HtrA高毒力株相对HtrA基因缺陷株具有更高的gbpC基因和蛋白表达量。结论:S.mutans gbpC的表达会受HtrA的影响。HtrA基因可能是S.mutans高致龋性的重要因素之一。展开更多
文摘Background The present study was undertaken to replicate the associations of representative polymorphisms in three genes （complement factor H （CFH）, complement factor B （BF） and HtrA serine peptidase 1 （HTRA1）） with exudative age-related macular degeneration （AMD） in a Han Chinese population, and to test if the modifiable environmental factors affect AMD susceptibility associated with different type of genotype in these genes. Methods An age, gender and ethnicity matched case-control study was conducted to genotype the representative single neucleotide polymorphisms （SNPs） loci including rs1061170 and rs1410996 in CFH, rs641153 and rs4151667 in BF and rs11200638 in HTRA1 gene in 144 exudative AMD patients and 126 normal controls using PCR-RFLP and direct resequencing. The demographic characteristics and behavioral risk factors were also recorded. Allelic and genotypic associations for individual SNP and joint associations with two loci were performed. The gene-gene and gene-environment interactions were analyzed using multivariate non-conditional Logistic regression analysis. Results The C risk allele frequencies for CFH Y402H （rs1061170） in cases and controls were 12.5% and 5.4% respectively, which were much lower than those in Caucasians （P 〈0.001）. Compared with TT homozygous genotype, the CT heterozygous genotype was positively associated with AMD with odds ratio （OR） of 3.23 （1.36-5.07）. However, the population attributable risk （PAR） of C allele was only 3.3% （1.4%-4.3%）. rs1410996 was also associated with AMD independent of Y402H. The ORs of exudative AMD for individuals carrying one copy risk allele and two copy risk alleles were 2.57 （1.21-5.45） and 4.76 （2.15-10.55） respectively, with correspondent PARs of 28.3% （2.0%-40.5%） and 38.2% （21.8%-45.4%）. rs11200636 in HTRA1 was another susceptible locus for AMD and the risk homozygotes were significantly susceptible for exudutive AMD （OR=3.98, 1.88-8.43） with PAR of 38.9% （24.3%-45.8%）
文摘子痫前期(preeclampsia)是妊娠期特有的疾病,目前仍是重要的公共卫生问题,是发展中国家孕产妇死亡的主要原因之一。研究表明子痫前期与细胞凋亡、血管痉挛、凝血系统激活、炎症反应等密切相关,其病理变化起始于胎盘浅着床和胎盘血管重铸障碍,导致胎盘组织的缺血缺氧,引起子痫前期的发生。HtrA(high temperature requirement A)系丝氨酸蛋白酶家族,该蛋白家族在细胞热休克、氧化应激、炎症、缺血再灌注和肿瘤等应激条件下,降解细胞胞质内错误折叠的蛋白质,参与细胞凋亡、关节炎、胚胎形成、神经变性和神经肌肉障碍以及癌症等的病理生理过程,研究表明其亚型与子痫前期密切相关。总结近年来与子痫前期有关的HtrA的研究进展及其介导的调控网络,对进一步了解子痫前期的发病机制及提供新的治疗途径有重要意义。
基金Supported by China Spark Program(2015GA620002)Key Technologies Research and Development Program of Hebei Province(14826613D)+2 种基金PhD Start-up Fund of Hebei Normal University of Science and Technology(2015YB002)Scientific and Technological Research Program of Institutions of Higher Education of Hebei Province(ZD2016067)the Fund of Qinhuangdao Academy of Agricultural Sciences(2014-04)
文摘To investigate the roles of HtrA protein in bacterial adaption to stresses, htra gene was cloned from an Escherichia coli strain isolated from silver fox, double digested, and ligated into the prokaryotic expression vector pET32a. Then, the recombinant plasmid pET32a-htra was transformed into E. coli BL21 competent cells. The over-expression of His-HtrA protein was induced by the addition of IPTG at a final concentration of 1 mmol/L, and verified by SDA-PAGE and Western blot. In addition, the protein was purified with Ni-NTA agarose, and used to immunize mice,and the polyclonal antibody specifically bound to E. coli HtrA protein. The results will provide a theoretical basis for studying the roles of HtrA protein and developing vaccines for fur-bearing animals against E. coli.
基金supported by Natural Science and Technology Fund of Hubei Povince(012726334)
文摘Objective:To explore HtrA1 gene expression aud its regulation in human gastric cancers.Methods:The HtrA1 mRNA levels were examined by QPCR analysis and coufirmed its expression with Northern blot analysis.The HtrA1 protein levels in all six gastric epithelial cell lines were investigated by Western blot analysis.Gene copy number was accessed and then sequenced the coding region from each mRNA in all six cell lines.The HtrA1 promoter region DNA methylation status was detected by using bisulfite sequeucing analysis.Effect of decitabine and TSA on HTRA1 expression in gastric cancer cell line was determined by RTPCR.Results:HIC analysis indicated that HtrA1 was highly expressed in normal epithelium,but dramatically down-regulated in gastric carcinoma tissues and variably expressed in tumor-adjacent tissues.HtrA1 gene expression was dramatically decreased in gastric carcinoma cells compared to nontumorigenic counterparts.The HtrA1 gene loss in any of the 4 breast cancer cell lines was not detected.Total 14 CpGs in this region were all methylated in gastric cancer cells,whereas two normal cells.GES-1 and HFI-145,were having several unmethylated cytosines in this region.HtrA1 showed as^Mr 44,000,Expression of HtrA1 protein was not observed in any of the four gastric caucer cell lines.BGC-823.MKN-45.SGC-7901and MKN-28.HtrA1 expression was observed in the HF1-145and GES-1 cell lines.Conclusions:The epigenetic silencing for HtrA1gene expression could provide a possible strategy for re-activating Htrt1 gene expression in gastric cancer cells.thus facilitating further investigation of HtrA1's role in chemotherapy.
基金sponsored by Chinese Academy of Sciences Visiting Professorship for Senior International Scientists(2010T1S6)by the DFG Cluster of Excellence"Inflammation at Interfaces"(EXC 306)by the Fonds der Chemischen Industrie
文摘Legionella pneumophila,the causative agent of Legionnaires' disease,has been recognized as a major health problem responsible for an estimated number of 15 000-30 000 cases of severe pneumonia per year in Germany alone.Despite of the high clinical relevance,many aspects of the intracellular life cycle of Legionella,especially details on interactions with host cells,are not well understood.Structural information on virulence proteins helps unravel basal pathogenicity mechanisms and is a prerequisite for the rational development of effective drug molecules.Here we discuss structures of three important virulence proteins of Legionella that have been determined in our laboratory.The structure of the macrophage infectivity potentiator(Mip) protein of Legionella pneumophila is the first of a novel subgroup within the family of FK506-binding protein(FKBP) peptidyl-prolyl cis/trans isomerases.On the basis of the Mip structure,promising antibacterial agents are being designed.Recently,structures of two equally exciting Legionella proteins have been reported.The ferrous iron transport protein FeoB is a transmembrane protein responsible for Fe2+ aquisition after entry of the pathogen into the host cell.The structure of the cytoplasmic domain of FeoB provides insights into the family of prokaryotic G proteins and allows a detailed comparison with structures of related FeoBs.Furthermore,the characterization of DegQ,a periplasmatic chaperone-protease involved in protein quality control represents an intriguing example of how enzymatic activity is regulated by oligomerization as well as by an intrinsic loop activation cascade,depending on subtle conformational rearrangements.
