1.Expression and purification of recombinant N-terminal peptide of human perforin
Weisong QIN ; Fangqiu LI ; Jianguo WU
Journal of Medical Postgraduates 2001;14(1):9-11
Objectives: To express and purify N-terminal peptide of human perforin(hPFP-N). Methods: The recombinant expressive plasmid pGEX-KG/hPFP-N was constructed and then introduced into a strain of E. coli BL21(DE3). Upon the induction of IPTG, GST/hPFP-N fusion protein was expressed. The expressed fusion protein was localized in inclusion bodies which could be solubilized by sonication after detergent lauroylsarcosine was added. The fusion protein was purified by affinity chromatography with glutathione agarose. After being cleaved by thrombin, GST and uncleaved fusion protein were removed by glutathione agarose beads once more, then purified recombinant rhPFP-N protein was obtained. Results and Conclusions: GST/hPFP-N fusion protein can be effectively expressed in E. coli and the protein hPFP-N was obtained after the purification process.
2.Establishment and clinical application of quantitative RT-PCR method for detecting perforin mRNA
Weisong QIN ; Fangqiu LI ; Jianguo WU
Chinese Journal of Clinical Laboratory Science 2001;19(3):137-139
Objective To establish the quantitative method for detecting perforin mRNA. Methods The competitive templates were prepared by restriction endonuclease method.The quantitative RT-PCR assay was established and used to detecting the perforin mRNA level in peripheral blood of tumor patients and healthy adults.Results Average perforin mRNA level of six healthy adults is 0.51±0.40 pmol/ml.The perforin mRNA level of patients with digestive tumors are significantly lower than that of healthy adults(P<0.01).But compared with healthy adults,no significant difference is showed in patients with breast cancer or leukemia.Conclusion The decreased perforin mRNA level probably attributes to the high incidence of cancers.Except for perforin-mediated cytolysis,additional effective mechanism against tumor cells maybe exist.
3.Soluble intracellular adhesion molecule-1 concentrations in the serum and peritoneal fluid from endometriosis patients
Qiuming ZHOU ; Yuanzhe WU ; Jinyan SHA ; Weisong QIN ;
Journal of Medical Postgraduates 2003;0(05):-
Objectives:To investigate the level of soluble intracellular adhesion molecule 1 (sICAM 1) concentrations in the serum and peritoneal fluid from the patients with or without endometriosis.To discuss the relationship within sICAM 1 and pelvic endometriosis. Methods:35 serum and 30 peritoneal fluid of patients with endometriosis (test group) and 26 serum and 4 peritoneal fluid without endometriosis (control group) were studied.Soluble intracellular adhesion molecule 1 levels were detected by an enzyme linked immunoassay(ELISA).The results were analysed by T test. Results: The serum concentration of sICAM 1 was significantly higher in test group than controll group. However,There was no significant difference in between two groups. Conclusions: These results suggest that sICAM 1 could play some role in the persistence of endometriotic lesions.
4.Expression of adhesion molecules in polymyositis
Wentao WU ; Weisong QIN ; Fuyuan SHAO ; Guanghui CHEN ; Xinfeng LIU ;
Journal of Medical Postgraduates 2003;0(12):-
Objective:To study the role of adhesion molecules in the pathogenesis of polymyositis. Methods:The abnormal expression of adhesion molecules on T cells in peripheral blood and muscle fibers from patients with myositis was analyzed by two colour immunofluoresence and RT PCR methods respectively. Results:The expression of adhesion molecules including lymphocyte function associated antigen 1(LFA 1 ),very late antigen 4(VLA 4) on T cells in peripheral blood and intercellular adhesion molecule l(ICAM 1) on muscle fibers from patients with myositis was markedly higher than that in the healthy control group. Conclusion: These findings suggested that adhesion molecules may be responsible for the migration of T cells and destraction of muscle fibers.
5.Mechanism of action of intestinal flora in hepatic encephalopathy and related treatment methods
Shenglan ZENG ; Chao LYU ; Weisong XIAO ; Xiaobin QIN ; Cong WU ; Dewen MAO
Journal of Clinical Hepatology 2020;36(10):2375-2379
At present, hepatic encephalopathy has a relatively high mortality and thus greatly affects patients’ quality of life. This article describes the changes of intestinal flora in patients with hepatic encephalopathy and analyzes the mechanism of action of intestinal flora in hepatic encephalopathy and related treatment methods. It is pointed out that the development of hepatic encephalopathy is closely associated with intestinal flora, and clinical treatment by regulating intestinal flora has achieved a marked effect in patients with hepatic encephalopathy. In the future, the research on intestinal flora in patients with hepatic encephalopathy can be deepened to provide better regimens for the treatment of hepatic encephalopathy.
6.Role of pyroptosis in liver diseases
Weisong XIAO ; Yingyu LE ; Shenglan ZENG ; Xiaobin QIN ; Cong WU ; Chengyu YA ; Dewen MAO
Journal of Clinical Hepatology 2020;36(12):2847-2850
As a novel form of programmed cell death different from cell necrosis, apoptosis, and autophagy discovered in recent years, pyroptosis is characterized by cell membrane rupture and release of cell contents and proinflammatory factors mediated by gasdermin, thus leading to cell death. Pyroptosis signaling pathways can be classified into classical pathways dependent on caspase-1 and non-classical pathways dependent on caspase-4/5/11; the activation of caspase-1 in classical pathways depends on the function of inflammasome, while the direct activation of caspase-4/5/11 is observed in non-classical pathways, which leads to the lysis of gasdermin D and induce the formation of membrane pores, the maturation and release of interleukin-1β and interleukin-18, and the rupture of cell membrane to cause pyroptosis. Latest research has shown that pyroptosis plays an important role in the development and progression of chronic liver diseases. This article introduces the mechanism of pyroptosis and summarizes the role of pyroptosis in the development and progression of nonalcoholic fatty liver disease, alcoholic liver disease, viral hepatitis, liver cirrhosis, and hepatocellular carcinoma, in order to provide new ideas and methods for the prevention and treatment of liver diseases in clinical practice.
7.Research advances in the pathogenesis of nonalcoholic fatty liver disease
Weisong XIAO ; Yingyu LE ; Shenglan ZENG ; Xiaobin QIN ; Cong WU ; Dewen MAO
Journal of Clinical Hepatology 2020;36(8):1874-1879
The development and progression of nonalcoholic fatty liver disease (NAFLD) have complex potential mechanisms. The traditional “two-hit” pathophysiological theory has been challenged, and in recent years, an increasing number of studies have been performed to investigate the interaction between insulin resistance, adipokines, and other unknown pathogenic factors in various organs. This article summarizes the factors of the liver, intestinal tract, hypothalamus, and extracellular cysts, as well as genetic factors, with an emphasis on the synergistic mechanism of action of the liver and extrahepatic organs in the pathogenesis of NAFLD, in order to provide a reference for obtaining new insights into NAFLD regulatory network and determining new targets for the prevention and treatment of NAFLD.