A new compound,3,5-dihydroxy-N-（3-nitrobenzylidene）benzohydrazide （DNBB） methanol solvate,was synthesized and characterized by single-crystal X-ray diffraction.DNBB crystallizes in a monoclinic system,space group ...A new compound,3,5-dihydroxy-N-（3-nitrobenzylidene）benzohydrazide （DNBB） methanol solvate,was synthesized and characterized by single-crystal X-ray diffraction.DNBB crystallizes in a monoclinic system,space group P2 1/c with a=8.387（4）,b=23.237（10）,c=9.133（3）,β=122.10（3）o,Z=4,V=1507.8（12） 3,D c=1.464 g/cm 3,F（000）=696.0,μ=0.116 mm-1,the final R=0.0480 and wR=0.1294.DNBB exhibited low toxicity on Caco-2 cell culture and produced significant inhibitory effect on the contraction of rat jejunal longitudinal smooth muscle （JLSM）.The mechanism of DNBB on JLSM was correlated to the stimulation of α and β adrenergic receptors since α receptor antagonist phentolamine and β receptor antagonist propranolol abolished the inhibitory effects of DNBB on the contraction of JLSM,respectively.展开更多
Objectives The cellular repressor of E1A-activated genes (CREG), a novel gene, was recently found to play a role in inhibiting cell growth and promoting cell differentiation. The purpose of this study was to obtain an...Objectives The cellular repressor of E1A-activated genes (CREG), a novel gene, was recently found to play a role in inhibiting cell growth and promoting cell differentiation. The purpose of this study was to obtain antibody against CREG protein and to study the expression of CREG protein in human internal thoracic artery cells (HITASY) which express different patterns of differentiation markers after serum withdrawal. Methods The open reading frame of CREG gene sequence was amplified by PCR and cloned into the pGEX-4T-1 vector. Glutathione-S-transferase (GST)-CREG fusion protein was expressed in E. Coli BL21 and purified from inclusion bodies by Sephacryl S-200 chromatography. Rabbits were immunized with the purified GST-CREG protein. Western blot examined with immunohistochemistry staining and the protein expression level was analyzed by Western blot in HITASY cells after serum removal. Results It was confirmed by using endonuclease digesting and DNA sequencing that the PCR product of CREG was correctly inserted into the vector. The GST-CREG protein was purified with gel filtration chromatography. Polyclonal antibody against GST-CREG was obtained from rabbits. CREG protein immunohistochemistry staining displayed a perinuclear distribution in the cytoplasm of HITASY cells. Results from Western blot suggested that comparing with the untreated cells upregulation of CREG polyclonal antibody against CREG was comfirmed. Using this antibody, the changes of CREG protein expression was observed in the process of phenotypic modulation of HITASY cells. These results provide basic understanding on the relationship of CREG gene with the cell phenotypic conversion.展开更多
目的:优化细辛多糖的提取工艺,研究细辛多糖对血管平滑肌细胞衰老的保护作用。方法:Box-Behnken响应面法优化多糖提取参数,AKTA explorer 100层析系统分级纯化多糖,利用形态学观察、MTT法、衰老β-半乳糖苷酶染色法评估细辛多糖对依托...目的:优化细辛多糖的提取工艺,研究细辛多糖对血管平滑肌细胞衰老的保护作用。方法:Box-Behnken响应面法优化多糖提取参数,AKTA explorer 100层析系统分级纯化多糖,利用形态学观察、MTT法、衰老β-半乳糖苷酶染色法评估细辛多糖对依托泊苷诱导大鼠主动脉平滑肌细胞衰老的保护作用。结果:细辛多糖的最佳提取条件为:提取温度90℃,提取时间2.37 h,液料比40∶1 mL/g,最大多糖得率为10.20%。细胞实验中,细辛多糖预处理明显减轻依托泊苷诱导所致的细胞衰老形态变化,提高细胞活力,下调衰老β-半乳糖苷酶活性水平。结论:细辛多糖的最佳提取工艺被确定,细辛多糖可明显抑制依托泊苷诱导的大鼠主动脉平滑肌细胞衰老和凋亡,对心血管疾病有保护作用。展开更多
基金supported by the National Natural Science Foundation of China (No. 30772601)
文摘A new compound,3,5-dihydroxy-N-（3-nitrobenzylidene）benzohydrazide （DNBB） methanol solvate,was synthesized and characterized by single-crystal X-ray diffraction.DNBB crystallizes in a monoclinic system,space group P2 1/c with a=8.387（4）,b=23.237（10）,c=9.133（3）,β=122.10（3）o,Z=4,V=1507.8（12） 3,D c=1.464 g/cm 3,F（000）=696.0,μ=0.116 mm-1,the final R=0.0480 and wR=0.1294.DNBB exhibited low toxicity on Caco-2 cell culture and produced significant inhibitory effect on the contraction of rat jejunal longitudinal smooth muscle （JLSM）.The mechanism of DNBB on JLSM was correlated to the stimulation of α and β adrenergic receptors since α receptor antagonist phentolamine and β receptor antagonist propranolol abolished the inhibitory effects of DNBB on the contraction of JLSM,respectively.
基金The work was supported by grant from The National Natural Sciences Foundation of China (No.30070280)
文摘Objectives The cellular repressor of E1A-activated genes (CREG), a novel gene, was recently found to play a role in inhibiting cell growth and promoting cell differentiation. The purpose of this study was to obtain antibody against CREG protein and to study the expression of CREG protein in human internal thoracic artery cells (HITASY) which express different patterns of differentiation markers after serum withdrawal. Methods The open reading frame of CREG gene sequence was amplified by PCR and cloned into the pGEX-4T-1 vector. Glutathione-S-transferase (GST)-CREG fusion protein was expressed in E. Coli BL21 and purified from inclusion bodies by Sephacryl S-200 chromatography. Rabbits were immunized with the purified GST-CREG protein. Western blot examined with immunohistochemistry staining and the protein expression level was analyzed by Western blot in HITASY cells after serum removal. Results It was confirmed by using endonuclease digesting and DNA sequencing that the PCR product of CREG was correctly inserted into the vector. The GST-CREG protein was purified with gel filtration chromatography. Polyclonal antibody against GST-CREG was obtained from rabbits. CREG protein immunohistochemistry staining displayed a perinuclear distribution in the cytoplasm of HITASY cells. Results from Western blot suggested that comparing with the untreated cells upregulation of CREG polyclonal antibody against CREG was comfirmed. Using this antibody, the changes of CREG protein expression was observed in the process of phenotypic modulation of HITASY cells. These results provide basic understanding on the relationship of CREG gene with the cell phenotypic conversion.
文摘目的:优化细辛多糖的提取工艺,研究细辛多糖对血管平滑肌细胞衰老的保护作用。方法:Box-Behnken响应面法优化多糖提取参数,AKTA explorer 100层析系统分级纯化多糖,利用形态学观察、MTT法、衰老β-半乳糖苷酶染色法评估细辛多糖对依托泊苷诱导大鼠主动脉平滑肌细胞衰老的保护作用。结果:细辛多糖的最佳提取条件为:提取温度90℃,提取时间2.37 h,液料比40∶1 mL/g,最大多糖得率为10.20%。细胞实验中,细辛多糖预处理明显减轻依托泊苷诱导所致的细胞衰老形态变化,提高细胞活力,下调衰老β-半乳糖苷酶活性水平。结论:细辛多糖的最佳提取工艺被确定,细辛多糖可明显抑制依托泊苷诱导的大鼠主动脉平滑肌细胞衰老和凋亡,对心血管疾病有保护作用。
