Recombinant E.coli JM109, containing pHZ1818 plasmid which included the fused gene encoding human interleukin 6(IL 6), expressed a fusion protein with glutathion S transferase(GST). The fusion protein existed both...Recombinant E.coli JM109, containing pHZ1818 plasmid which included the fused gene encoding human interleukin 6(IL 6), expressed a fusion protein with glutathion S transferase(GST). The fusion protein existed both in the supernatant and inside the bacterial cell,but the insoluble protein had no biological activity and could not be refolded. The rotative speed of the shaker and the temperature of induction were optimized to maximize the expression of the soluble fusion protein. From the supernatant of the cell sonicates Glutathion Sephrose 4B affinity column chromatography was employed to isolate the fusion protein which could be purified to >80 0 0 in a single step. The yield of soluble GST IL 6 was about 10 mg per liter culture. The GST was site specifically cloven by 6 hours of treatment with thrombin and from the thrombin digest mixture IL 6 was purified by Q high performance ion exchange chromatography. From 1 liter of E.coli culture 2 mg refined IL 6 was obtained. The purified IL 6 had a purity of more than 95 0 0 and a biological activity of 1.02×10 8 IU/mg.展开更多
Objective:To investigate the effect of IL-6 gene transfe r into human cord blood hematopoietic stem cells on the production of megakaryocy t ic progenitors.Methods:IL-6 gene was transfected into human c o rd blood CD3...Objective:To investigate the effect of IL-6 gene transfe r into human cord blood hematopoietic stem cells on the production of megakaryocy t ic progenitors.Methods:IL-6 gene was transfected into human c o rd blood CD34+ cells using a retrovirus vector with the aid of recombinant fibro nectin fragments in the presence of a cocktail of cytokines (SCF,IL-6,sIL-6R,F L,and TPO).Colony-forming units-megakaryocyte (CFU-MK) assays were performed as IL-6 gene transduced CD34+ cells were incubated alone or in combination with I L -3 or sIL-6R, controlled with neoR gene transduced CD34+ cells.Result s :IL-6 alone or sIL-6R alone stimulated few CFU-MK colonies,the addit ion of sIL-6R to IL-6 gene transduced CD34+ cells significantly enhanced the prod uction of CFU-MK colonies.IL-6 gene transduced CD34+ cells showed a modest syn er gistic effect with IL-3.Conclusion:These results suggest that IL-6 gene transfer may protect patients from chemotherapy-induced thrombocytop en ia.展开更多
目的探讨人转运甲状腺素蛋白(transthyretin,TTR)、人凝聚素(human clusterin,CLU)、白细胞介素-6(interleukin-6,IL-6)与脑卒中后继发认知功能障碍的相关性及预后预测价值。方法选取高邮市中医医院收治的500例脑卒中患者,根据简易智能...目的探讨人转运甲状腺素蛋白(transthyretin,TTR)、人凝聚素(human clusterin,CLU)、白细胞介素-6(interleukin-6,IL-6)与脑卒中后继发认知功能障碍的相关性及预后预测价值。方法选取高邮市中医医院收治的500例脑卒中患者,根据简易智能精神状态检查量表(mini-mental state examination,MMSE)得分,分为认知功能障碍组180例和非认知功能障碍组320例,另选取200名健康志愿者作为对照组,对比3组受检者TTR、CLU、IL-6表达水平,并分析与脑卒中后继发认知功能障碍的相关性。随访患者预后情况,根据180例脑卒中后继发认知功能障碍患者治疗后的MMSE得分分为预后良好组110例和预后不良组70例,对比2组患者临床一般情况与TTR、CLU、IL-6表达水平,并用Logistic回归分析分析这些指标对卒中后继发认知功能障碍的预后预测价值。结果认知功能障碍组TTR、IL-6高于非认知功能障碍组和对照组,CLU低于非认知功能障碍组和对照组(P<0.05);Spearman相关分析结果显示,TTR、IL-6与脑卒中后继发认知功能障碍呈正相关,与CLU呈负相关(P>0.05);预后良好组与预后不良组患者体质量指数,是否首发,TTR、CLU、IL-6水平对比差异有统计学意义(P<0.05);Logistic回归分析显示,TTR、CLU、IL-6为脑卒中后继发认知功能障碍的预后独立影响因素(P<0.05)。结论血清TTR、CLU、IL-6水平不仅与脑卒中后继发认知功能障碍具有明显相关性,还为其预后独立影响因素。因此,临床上可考虑将TTR、CLU、IL-6三者作为脑卒中后认知功能障碍的诊断及预后预测指标。展开更多
文摘Recombinant E.coli JM109, containing pHZ1818 plasmid which included the fused gene encoding human interleukin 6(IL 6), expressed a fusion protein with glutathion S transferase(GST). The fusion protein existed both in the supernatant and inside the bacterial cell,but the insoluble protein had no biological activity and could not be refolded. The rotative speed of the shaker and the temperature of induction were optimized to maximize the expression of the soluble fusion protein. From the supernatant of the cell sonicates Glutathion Sephrose 4B affinity column chromatography was employed to isolate the fusion protein which could be purified to >80 0 0 in a single step. The yield of soluble GST IL 6 was about 10 mg per liter culture. The GST was site specifically cloven by 6 hours of treatment with thrombin and from the thrombin digest mixture IL 6 was purified by Q high performance ion exchange chromatography. From 1 liter of E.coli culture 2 mg refined IL 6 was obtained. The purified IL 6 had a purity of more than 95 0 0 and a biological activity of 1.02×10 8 IU/mg.
文摘Objective:To investigate the effect of IL-6 gene transfe r into human cord blood hematopoietic stem cells on the production of megakaryocy t ic progenitors.Methods:IL-6 gene was transfected into human c o rd blood CD34+ cells using a retrovirus vector with the aid of recombinant fibro nectin fragments in the presence of a cocktail of cytokines (SCF,IL-6,sIL-6R,F L,and TPO).Colony-forming units-megakaryocyte (CFU-MK) assays were performed as IL-6 gene transduced CD34+ cells were incubated alone or in combination with I L -3 or sIL-6R, controlled with neoR gene transduced CD34+ cells.Result s :IL-6 alone or sIL-6R alone stimulated few CFU-MK colonies,the addit ion of sIL-6R to IL-6 gene transduced CD34+ cells significantly enhanced the prod uction of CFU-MK colonies.IL-6 gene transduced CD34+ cells showed a modest syn er gistic effect with IL-3.Conclusion:These results suggest that IL-6 gene transfer may protect patients from chemotherapy-induced thrombocytop en ia.
文摘目的探讨人转运甲状腺素蛋白(transthyretin,TTR)、人凝聚素(human clusterin,CLU)、白细胞介素-6(interleukin-6,IL-6)与脑卒中后继发认知功能障碍的相关性及预后预测价值。方法选取高邮市中医医院收治的500例脑卒中患者,根据简易智能精神状态检查量表(mini-mental state examination,MMSE)得分,分为认知功能障碍组180例和非认知功能障碍组320例,另选取200名健康志愿者作为对照组,对比3组受检者TTR、CLU、IL-6表达水平,并分析与脑卒中后继发认知功能障碍的相关性。随访患者预后情况,根据180例脑卒中后继发认知功能障碍患者治疗后的MMSE得分分为预后良好组110例和预后不良组70例,对比2组患者临床一般情况与TTR、CLU、IL-6表达水平,并用Logistic回归分析分析这些指标对卒中后继发认知功能障碍的预后预测价值。结果认知功能障碍组TTR、IL-6高于非认知功能障碍组和对照组,CLU低于非认知功能障碍组和对照组(P<0.05);Spearman相关分析结果显示,TTR、IL-6与脑卒中后继发认知功能障碍呈正相关,与CLU呈负相关(P>0.05);预后良好组与预后不良组患者体质量指数,是否首发,TTR、CLU、IL-6水平对比差异有统计学意义(P<0.05);Logistic回归分析显示,TTR、CLU、IL-6为脑卒中后继发认知功能障碍的预后独立影响因素(P<0.05)。结论血清TTR、CLU、IL-6水平不仅与脑卒中后继发认知功能障碍具有明显相关性,还为其预后独立影响因素。因此,临床上可考虑将TTR、CLU、IL-6三者作为脑卒中后认知功能障碍的诊断及预后预测指标。
