Porcine proliferative enteropathy(PPE),an important infectious disease in pig production caused by an obligate intracellular bacterium Lawsonia intracellularis,is commonly associated with diarrhea and reduced weight g...Porcine proliferative enteropathy(PPE),an important infectious disease in pig production caused by an obligate intracellular bacterium Lawsonia intracellularis,is commonly associated with diarrhea and reduced weight gain in growing pigs widespread.An accurate method for detecting L.intracellularis is particularly important for preventing and controlling PPE.Heat shock protein 60(Hsp60)is an immunodominant bacterial antigen found in all eukaryotic and prokaryotic organisms.Thus,the purpose of the current investigation was to produce a novel L.intracellularis Hsp60 monoclonal antibody(mAb)useful for immunodiagnostics.Three hybridomas secreted anti-Hsp60 termed 3E5,4E2,and 9G6 were generated,and the titers of ascitic fluids of 3E5,4E2,9G6 were 1:1024000,1:2048000 and 1:2048000,respectively.The Western blotting analysis demonstrated that recombinant Hsp60(rHsp60)was recognized by mAbs 3E5,4E2 and 9G6.Subsequently,analyses of specificity showed all the mAbs were highly specific to L.intracellularis while could not significantly react with other enteric bacteria commonly found in the ileum of pigs,such as Escherichia coli,Salmonella Choleraesuis,Salmonella Typhimurium,and Brachyspira hyodysenteriae.Furthermore,the mAbs were useful for detecting L.intracellularis in the infected monolayer cells and histological sections of the ileum from PPE-affected pigs.Our research will provide a foundation for the development of immunological diagnostic tests.展开更多
Heat shock proteins 10/60(hsp10/60)are a family of conserved ubiquitously expressed heat shock proteins which are produced by cells in response to exposure to stressful conditions.Besides the chaperone and housekeepin...Heat shock proteins 10/60(hsp10/60)are a family of conserved ubiquitously expressed heat shock proteins which are produced by cells in response to exposure to stressful conditions.Besides the chaperone and housekeeping functions,they are also known to be involved in immune response during bacterial infection.In this study,we identified and annotated 10 hsp10/60 genes through bioinformatic analysis in Japanese flounder(Paralichthys olivaceus).Among them one member of hsp10(hspe)family and nine members of hsp60(hspd)family were identified.Phylogenetic and selection pressure analysis showed that the hsp10/60 genes were evolutionarily constrained and their function was conserved.Besides,hsp10/60 genes were involved in different embryonic and larval stages and acted as the sentinel role in an unchallenged organism.In addition,we also observed the expression patterns of hsp10/60 genes after Edwardsiella tarda infection,for the first time in Japanese flounder.Eight out of 10 genes were differentially expressed after bacterial challenges,the significantly regulated expressions of flounder hsp10/60 genes after bacterial infections suggested their involvement in immune response in flounder.Our results provide valuable information for clarifying the evolutionary relationship,and early insights of the immune functions of hsp10/60 genes in Japanese flounder.展开更多
目的:探讨血清anti-热休克蛋白60(Heat shock protein 60,HSP60)、睾酮水平对多囊卵巢综合征(PCOS)合并不孕的预测价值。方法:2018年2月至2021年2月选择在本院诊治的多囊卵巢综合征合并不孕患者88例作为研究组,同期选择在本院体检中心...目的:探讨血清anti-热休克蛋白60(Heat shock protein 60,HSP60)、睾酮水平对多囊卵巢综合征(PCOS)合并不孕的预测价值。方法:2018年2月至2021年2月选择在本院诊治的多囊卵巢综合征合并不孕患者88例作为研究组,同期选择在本院体检中心进行孕前体检的88例健康妇女作为对照组。检测研究组与对照组血清anti-HSP60、睾酮水平并进行多因素与预测价值分析。结果:研究组与对照组的年龄、体重指数、初潮年龄、腰围、空腹血糖、低密度脂蛋白胆固醇、高密度脂蛋白胆固醇、总胆固醇、甘油三酯,对比无显著差异(P>0.05)。研究组的血清anti-HSP60、睾酮水平高于对照组(P<0.05)。多因素Logistic回归分析显示,血清anti-HSP60、睾酮是多囊卵巢综合征合并不孕发生的独立危险因素(P<0.05)。ROC曲线分析显示血清anti-HSP60、睾酮预测PCOS合并不孕症的曲线下面积分别为0.782、0.813,预测敏感性与特异性都超过了70.0%。结论:血清anti-HSP60、睾酮水平对多囊卵巢综合征合并不孕患者中呈现高表达水平,也是导致不孕发生的重要因素,对于多囊卵巢综合征合并不孕的预测也具有重要价值。展开更多
AIM: To determine the overall prevalence of H pylori and CagA positive H pylori infection and the prevalence of other bacterial and viral causes of chronic infection in patients with coronary heart disease (CHD), and ...AIM: To determine the overall prevalence of H pylori and CagA positive H pylori infection and the prevalence of other bacterial and viral causes of chronic infection in patients with coronary heart disease (CHD), and the potential role of anti-heat-shock protein 60 (Hsp60) anti- body response to these proteins in increasing the risk of CHD development. METHODS: Eighty patients with CHD and 160 controls were employed. We also compared the levels of anti- heat-shock protein 60 (Hsp60) antibodies in the two groups. The H pylori infection and the CagA status were determined serologically, using commercially available enzyme-linked immunosorbent assays (ELISA), and a Western blotting method developed in our laboratory. Systemic antibodies to Hsp60 were determined by a sandwich ELISA, using a polyclonal antibody to Hsp60 to sensitise polystyrene plates and a commercially available human Hsp60 as an antigen. RESULTS: The overall prevalence of H pylori infec- tion was 78.7% (n = 63) in patients and 76.2% (n = 122) in controls (P = 0.07). Patients infected by CagA- positive (CagA+) H pylori strains were 71.4% (n = 45) vs 52.4% of infected controls (P = 0.030, OR = 2.27). Sys-temic levels of IgG to Hsp60 were increased in H pylori- negative patients compared with uninfected controls (P < 0.001) and CagA-positive infected patients compared with CagA-positive infected controls (P = 0.007). CONCLUSION: CagA positive H pylori infection may concur to the development of CHD; high levels of anti- Hsp60 antibodies may constitute a marker and/or a con- comitant pathogenic factor of the disease.展开更多
several heat shock proteins have been investigated in relation to tooth development, no available information is available about the spatial and temporal expression pattern of heat shock protein 60 （Hsp 60）. To char...several heat shock proteins have been investigated in relation to tooth development, no available information is available about the spatial and temporal expression pattern of heat shock protein 60 （Hsp 60）. To characterize Hsp 60 expression in the structures of the developing tooth germ, we used Western blotting, immunohistochemistry and in situ hybridization. Hsp 60 was present in high amounts in the inner and outer enamel epithelia, enamel knot （EK） and stratum intermedium （SI）. Hsp 60 also appeared in odontoblasts beginning in the bell stage. To obtain data on the possible effect of Hsp 60 on isolated lower incisors from mice, we performed in vitro culturing. To investigate the effect of exogenous Hsp 60 on the cell cycle during culturing, we used the 5-bromo-2- deoxyuridine （BrdU） incorporation test on dental cells. Exogenously administered Hsp 60 caused bluntness at the apical part of the 16.5-day-old tooth germs, but it did not influence the proliferation rate of dental cells. We identified the expression of Hsp 60 in the developing tooth germ, which was present in high concentrations in the inner and outer enamel epithelia, EK, SI and odontoblasts. High concentration of exogenous Hsp 60 can cause abnormal morphology of the tooth germ, but it did not influence the proliferation rate of the dental cells. Our results suggest that increased levels of Hsp 60 may cause abnormalities in the morphological development of the tooth germ and support the data on the significance of Hsp during the developmental processes.展开更多
目的探讨利拉鲁肽联合达格列净对老年2型糖尿病患者血糖波动、服药安全性及热休克蛋白60(HSP60)、Nesfatin-1水平的影响。方法选取我院2019年5月至2021年3月收治的96例老年2型糖尿病患者,根据入院顺序将其分为对照组和观察组,各48例。...目的探讨利拉鲁肽联合达格列净对老年2型糖尿病患者血糖波动、服药安全性及热休克蛋白60(HSP60)、Nesfatin-1水平的影响。方法选取我院2019年5月至2021年3月收治的96例老年2型糖尿病患者,根据入院顺序将其分为对照组和观察组,各48例。对照组采用达格列净治疗,观察组采用利拉鲁肽联合达格列净治疗。比较两组的治疗效果。结果治疗后,观察组的空腹血糖(FPG)、餐后2 h血糖(2 h PG)、总胆固醇(TC)及甘油三酯(TG)水平均低于对照组(P<0.05)。治疗后,观察组的平均血糖(MBG)、血糖水平标准差(SDBG)、最大血糖波动幅度(LAGE)及餐后血糖波动幅度(PPGE)水平均低于对照组(P<0.05)。治疗后,观察组的脂联素、一氧化氮(NO)水平高于对照组,内皮素(ET)水平低于对照组(P<0.05)。两组的胃肠道不适、头晕、低血糖、皮疹发生率及不良反应总发生率比较,差异无统计学意义(P>0.05)。治疗后,观察组的HSP60及Nesfatin-1水平均低于对照组(P<0.05)。结论采用利拉鲁肽联合达格列净治疗老年2型糖尿病患者有助于降低血糖和血脂水平,减轻血糖波动,改善脂联素和血管内皮功能指标水平,降低HSP60、Nesfatin-1水平,且不会增加不良反应的发生。展开更多
基金supported by the National Key Research and Development Program of China(2021YFD1800400)the National Natural Science Foundation of China(31872480)+1 种基金the Jiangsu Agriculture Science and Technology Innovation Fund,China(CX(19)2020)the Priority Academic Program Development of Jiangsu Higher Education Institutions,China(PAPD).
文摘Porcine proliferative enteropathy(PPE),an important infectious disease in pig production caused by an obligate intracellular bacterium Lawsonia intracellularis,is commonly associated with diarrhea and reduced weight gain in growing pigs widespread.An accurate method for detecting L.intracellularis is particularly important for preventing and controlling PPE.Heat shock protein 60(Hsp60)is an immunodominant bacterial antigen found in all eukaryotic and prokaryotic organisms.Thus,the purpose of the current investigation was to produce a novel L.intracellularis Hsp60 monoclonal antibody(mAb)useful for immunodiagnostics.Three hybridomas secreted anti-Hsp60 termed 3E5,4E2,and 9G6 were generated,and the titers of ascitic fluids of 3E5,4E2,9G6 were 1:1024000,1:2048000 and 1:2048000,respectively.The Western blotting analysis demonstrated that recombinant Hsp60(rHsp60)was recognized by mAbs 3E5,4E2 and 9G6.Subsequently,analyses of specificity showed all the mAbs were highly specific to L.intracellularis while could not significantly react with other enteric bacteria commonly found in the ileum of pigs,such as Escherichia coli,Salmonella Choleraesuis,Salmonella Typhimurium,and Brachyspira hyodysenteriae.Furthermore,the mAbs were useful for detecting L.intracellularis in the infected monolayer cells and histological sections of the ileum from PPE-affected pigs.Our research will provide a foundation for the development of immunological diagnostic tests.
基金This work was supported by the National Key Research and Development Program of China(No.2018YFD0900601)the Natural Science Foundation of Shandong Province(No.ZR2017MC072).
文摘Heat shock proteins 10/60(hsp10/60)are a family of conserved ubiquitously expressed heat shock proteins which are produced by cells in response to exposure to stressful conditions.Besides the chaperone and housekeeping functions,they are also known to be involved in immune response during bacterial infection.In this study,we identified and annotated 10 hsp10/60 genes through bioinformatic analysis in Japanese flounder(Paralichthys olivaceus).Among them one member of hsp10(hspe)family and nine members of hsp60(hspd)family were identified.Phylogenetic and selection pressure analysis showed that the hsp10/60 genes were evolutionarily constrained and their function was conserved.Besides,hsp10/60 genes were involved in different embryonic and larval stages and acted as the sentinel role in an unchallenged organism.In addition,we also observed the expression patterns of hsp10/60 genes after Edwardsiella tarda infection,for the first time in Japanese flounder.Eight out of 10 genes were differentially expressed after bacterial challenges,the significantly regulated expressions of flounder hsp10/60 genes after bacterial infections suggested their involvement in immune response in flounder.Our results provide valuable information for clarifying the evolutionary relationship,and early insights of the immune functions of hsp10/60 genes in Japanese flounder.
文摘目的:探讨血清anti-热休克蛋白60(Heat shock protein 60,HSP60)、睾酮水平对多囊卵巢综合征(PCOS)合并不孕的预测价值。方法:2018年2月至2021年2月选择在本院诊治的多囊卵巢综合征合并不孕患者88例作为研究组,同期选择在本院体检中心进行孕前体检的88例健康妇女作为对照组。检测研究组与对照组血清anti-HSP60、睾酮水平并进行多因素与预测价值分析。结果:研究组与对照组的年龄、体重指数、初潮年龄、腰围、空腹血糖、低密度脂蛋白胆固醇、高密度脂蛋白胆固醇、总胆固醇、甘油三酯,对比无显著差异(P>0.05)。研究组的血清anti-HSP60、睾酮水平高于对照组(P<0.05)。多因素Logistic回归分析显示,血清anti-HSP60、睾酮是多囊卵巢综合征合并不孕发生的独立危险因素(P<0.05)。ROC曲线分析显示血清anti-HSP60、睾酮预测PCOS合并不孕症的曲线下面积分别为0.782、0.813,预测敏感性与特异性都超过了70.0%。结论:血清anti-HSP60、睾酮水平对多囊卵巢综合征合并不孕患者中呈现高表达水平,也是导致不孕发生的重要因素,对于多囊卵巢综合征合并不孕的预测也具有重要价值。
基金Supported by a grant from the University of Siena, PAR 2004 "H pylori infection, hosts’ aplotypes of inflammatory cytokines and the risk of ischemic heart disease"
文摘AIM: To determine the overall prevalence of H pylori and CagA positive H pylori infection and the prevalence of other bacterial and viral causes of chronic infection in patients with coronary heart disease (CHD), and the potential role of anti-heat-shock protein 60 (Hsp60) anti- body response to these proteins in increasing the risk of CHD development. METHODS: Eighty patients with CHD and 160 controls were employed. We also compared the levels of anti- heat-shock protein 60 (Hsp60) antibodies in the two groups. The H pylori infection and the CagA status were determined serologically, using commercially available enzyme-linked immunosorbent assays (ELISA), and a Western blotting method developed in our laboratory. Systemic antibodies to Hsp60 were determined by a sandwich ELISA, using a polyclonal antibody to Hsp60 to sensitise polystyrene plates and a commercially available human Hsp60 as an antigen. RESULTS: The overall prevalence of H pylori infec- tion was 78.7% (n = 63) in patients and 76.2% (n = 122) in controls (P = 0.07). Patients infected by CagA- positive (CagA+) H pylori strains were 71.4% (n = 45) vs 52.4% of infected controls (P = 0.030, OR = 2.27). Sys-temic levels of IgG to Hsp60 were increased in H pylori- negative patients compared with uninfected controls (P < 0.001) and CagA-positive infected patients compared with CagA-positive infected controls (P = 0.007). CONCLUSION: CagA positive H pylori infection may concur to the development of CHD; high levels of anti- Hsp60 antibodies may constitute a marker and/or a con- comitant pathogenic factor of the disease.
基金supported by the European Union and the State of Hungary and co-financed by the European Social Fund in the framework of TA′MOP 4.2.4.A/2-11-12012-0001‘National Excellence Program’supported by a Ja′nos Bolyai fellowship from the Hungarian Academy of Science
文摘several heat shock proteins have been investigated in relation to tooth development, no available information is available about the spatial and temporal expression pattern of heat shock protein 60 （Hsp 60）. To characterize Hsp 60 expression in the structures of the developing tooth germ, we used Western blotting, immunohistochemistry and in situ hybridization. Hsp 60 was present in high amounts in the inner and outer enamel epithelia, enamel knot （EK） and stratum intermedium （SI）. Hsp 60 also appeared in odontoblasts beginning in the bell stage. To obtain data on the possible effect of Hsp 60 on isolated lower incisors from mice, we performed in vitro culturing. To investigate the effect of exogenous Hsp 60 on the cell cycle during culturing, we used the 5-bromo-2- deoxyuridine （BrdU） incorporation test on dental cells. Exogenously administered Hsp 60 caused bluntness at the apical part of the 16.5-day-old tooth germs, but it did not influence the proliferation rate of dental cells. We identified the expression of Hsp 60 in the developing tooth germ, which was present in high concentrations in the inner and outer enamel epithelia, EK, SI and odontoblasts. High concentration of exogenous Hsp 60 can cause abnormal morphology of the tooth germ, but it did not influence the proliferation rate of the dental cells. Our results suggest that increased levels of Hsp 60 may cause abnormalities in the morphological development of the tooth germ and support the data on the significance of Hsp during the developmental processes.
文摘目的探讨利拉鲁肽联合达格列净对老年2型糖尿病患者血糖波动、服药安全性及热休克蛋白60(HSP60)、Nesfatin-1水平的影响。方法选取我院2019年5月至2021年3月收治的96例老年2型糖尿病患者,根据入院顺序将其分为对照组和观察组,各48例。对照组采用达格列净治疗,观察组采用利拉鲁肽联合达格列净治疗。比较两组的治疗效果。结果治疗后,观察组的空腹血糖(FPG)、餐后2 h血糖(2 h PG)、总胆固醇(TC)及甘油三酯(TG)水平均低于对照组(P<0.05)。治疗后,观察组的平均血糖(MBG)、血糖水平标准差(SDBG)、最大血糖波动幅度(LAGE)及餐后血糖波动幅度(PPGE)水平均低于对照组(P<0.05)。治疗后,观察组的脂联素、一氧化氮(NO)水平高于对照组,内皮素(ET)水平低于对照组(P<0.05)。两组的胃肠道不适、头晕、低血糖、皮疹发生率及不良反应总发生率比较,差异无统计学意义(P>0.05)。治疗后,观察组的HSP60及Nesfatin-1水平均低于对照组(P<0.05)。结论采用利拉鲁肽联合达格列净治疗老年2型糖尿病患者有助于降低血糖和血脂水平,减轻血糖波动,改善脂联素和血管内皮功能指标水平,降低HSP60、Nesfatin-1水平,且不会增加不良反应的发生。
