Objective: To discuss the relationship between hypercholesterolemic disease and the functional and structural changes of Sphincter of Oddi (SO) by the study of effect of Cholesterol Liposome (CL) on structural and qua...Objective: To discuss the relationship between hypercholesterolemic disease and the functional and structural changes of Sphincter of Oddi (SO) by the study of effect of Cholesterol Liposome (CL) on structural and quantitative changes of SO cells. Methods: Rabbit SO was isolated for primary cell culture and subculture. After subcultured with different concentration of CL culture medium for 20 h, the structural and quantitative changes of SO cells were analyzed and detected by MTT-test, flow cytometer (FCM), electronic microscope and electrophoresis technique respectively. Results: CL contributed a prominent stimulus to SO cells proliferation at middle concentration (<0. 5 - 0. 8 mg/ml), which could be confirmed by FCM analysis which indicated the number of SO cells in S-phase increasing remarkably; however, high concentration of CL inhibited SO cells' proliferation (>1. 0 mg/ml) and induced apoptosis of SO cells. Swelled mitochondria and dilated endoplasmic reticulum as well as disjoined and diminished microfilaments were found in SO cells by electronic microscopy. The content of SO cells actin decreased with the increment of cholesterol concentration. There was a significant difference of actin content between CL groups and control group (P<0. 05). Conclusion: CL may change SO cell membrane's function, organelle's structure and especially the quantity and configuration of microfilaments, at the same time, CL at different concentration can induce changes of SO cells cycle and lead to different changes in the number of SO cells.展开更多
Objective: To amplify human era (Hera) gene, then express it in E.coli. Methods: Human era gene, after amplified by PCR and identified by sequencing, was inserted into the expression vector pGEX-4T3 in which exogenous...Objective: To amplify human era (Hera) gene, then express it in E.coli. Methods: Human era gene, after amplified by PCR and identified by sequencing, was inserted into the expression vector pGEX-4T3 in which exogenous gene was controlled by Ptac promoter. The recombinant plasmid pGEX-Hera was transformed into DH5 (and induced with IPTG chemically. Results: The human era gene was amplified and the sequence was correct. When the bacteria with pGEX-Hera was induced, an anticipated 65 000 protein band appeared on SDS-PAGE gel and amounted to 23% of total bacterial protein. Conclusion: The human era gene has been successfully amplified and efficiently expressed in E.coli.展开更多
文摘Objective: To discuss the relationship between hypercholesterolemic disease and the functional and structural changes of Sphincter of Oddi (SO) by the study of effect of Cholesterol Liposome (CL) on structural and quantitative changes of SO cells. Methods: Rabbit SO was isolated for primary cell culture and subculture. After subcultured with different concentration of CL culture medium for 20 h, the structural and quantitative changes of SO cells were analyzed and detected by MTT-test, flow cytometer (FCM), electronic microscope and electrophoresis technique respectively. Results: CL contributed a prominent stimulus to SO cells proliferation at middle concentration (<0. 5 - 0. 8 mg/ml), which could be confirmed by FCM analysis which indicated the number of SO cells in S-phase increasing remarkably; however, high concentration of CL inhibited SO cells' proliferation (>1. 0 mg/ml) and induced apoptosis of SO cells. Swelled mitochondria and dilated endoplasmic reticulum as well as disjoined and diminished microfilaments were found in SO cells by electronic microscopy. The content of SO cells actin decreased with the increment of cholesterol concentration. There was a significant difference of actin content between CL groups and control group (P<0. 05). Conclusion: CL may change SO cell membrane's function, organelle's structure and especially the quantity and configuration of microfilaments, at the same time, CL at different concentration can induce changes of SO cells cycle and lead to different changes in the number of SO cells.
基金research grant from the National NaturalScience Foundation of China (No. 39870380, 39670006) the ScienceFoundation of PLA (
文摘Objective: To amplify human era (Hera) gene, then express it in E.coli. Methods: Human era gene, after amplified by PCR and identified by sequencing, was inserted into the expression vector pGEX-4T3 in which exogenous gene was controlled by Ptac promoter. The recombinant plasmid pGEX-Hera was transformed into DH5 (and induced with IPTG chemically. Results: The human era gene was amplified and the sequence was correct. When the bacteria with pGEX-Hera was induced, an anticipated 65 000 protein band appeared on SDS-PAGE gel and amounted to 23% of total bacterial protein. Conclusion: The human era gene has been successfully amplified and efficiently expressed in E.coli.
