1.Factors affecting establishment of rodent alcohol drinking models and recent advances
Xin LIU ; Qianqian ZHANG ; Shengxia WANG ; Wangxin ZHANG ; Hanting ZHANG
Chinese Journal of Pharmacology and Toxicology 2017;31(6):607-614
Alcohol use disorder (AUD) is one of the main factors that threaten human health and life.WHO data show that about 3.3 million people die each year worldwide due to AUD.AUD has become a global public health concern,but its neurobiological and behavioral mechanisms remain unclear.Therefore,it is particularly urgent to develop valid and reliable animal models to investigate the underlying mechanisms of AUD.This review summarizes the animal species and modeling methods of AUD in order to explore the potential influence on the deviation of modeling in alcohol intake and provide helpful inputs to behavioral research of AUD.
2.Effects of hydrogen sulfide on high glucose-induced injury of mouse glo-merular podocytes
Ye LIU ; Li PENG ; Shengxia LU ; Yuejuan DU ; Yuantao LIU ; Yuqin FU
Chinese Journal of Pathophysiology 2014;33(4):670-674
AIM:To investigate the effect of hydrogen sulfide ( H2 S) on high glucose ( HG)-induced injury of the mouse podocyte cell line MPC5.METHODS: The cultured MPC5 cells were randomly divided into 4 groups: HG group, normal glucose (NG) group, NG+DL-propargylglycine (PPG) group, and HG+NaHS group.After treated for a certain time, the cells were collected for further detection .The expression of zonula occludens-2 (ZO-2), nephrin,β-cate-nin and cystathionine γ-lyase ( CSE) was determined by Western blotting .RESULTS:High glucose significantly reduced the expression of nephrin, ZO-2 and CSE (P<0.05), while the level of β-catenin was elevated obviously (P<0.05), all in a time-dependent manner.NG+PPG inhibited the levels of ZO-2 and nephrin significantly (P<0.05), and increased the level of β-catenin (P<0.05), all in a PPG concentration-dependent manner.HG+NaHS induced a more significant increase in the levels of ZO-2 and nephrin as compared with HG group (P<0.01), whereas a severe reduction of β-cate-nin in HG+NaHS group was observed as compared with HG group .Compared with NG group , the expression of ZO-2 and nephrin was decreased obviously , and the level of β-catenin was increased in HG +NaHS group.CONCLUSION:Down-regulation of CSE contributes to hyperglycemia-induced podocyte injury .Exogenous H 2 S protects against hyperglycemia-in-duced podocyte injury , possibly through up-regulation of ZO-2 and subsequent suppression of Wnt/β-catenin pathway .
3.Phosphatidylserine exposure in red blood cell in sepsis patients
Guoguang MA ; Hongxiang LIU ; Changfang LIU ; Wanhua YANG ; Hongze ZHANG ; Shengxia WAN ; Bin SHI
International Journal of Biomedical Engineering 2013;36(5):270-273,后插1
Objective To investigate the morphologic change and phosphatidylserine (PS) exposure of erythrocytes in sepsis patients.Methods 30 healthy volunteers (control group)and 30 sepsis patients were enrolled in this study and were collected venous sampling.Monitoring included Wright's staining blood smear test,erythrocyte aggregation index and the ratio of PS exposure of erythrocytes.A flow-cytometric assay based on FITC-Annexin V was used to measure the PS exposure of erythrocytes.Results The morphological changes of red blood cells included acanthocyte,lachrymiform,rouleaux,spherocyte in sepsis patients,and the peripheral blood erythrocyte aggregation and aggregation index were significantly higher than that of the healthy control group (P<0.05).The percentage of PS exposure of erythrocytes in sepsis patients were significantly higher than that of healthy volunteers (P<0.001).Conclusion The PS exposure of erythrocytes were significantly higher in sepsis patients,and the morphology of red blood cells is obvious abnormal.
4.Prokaryotic expression, purification and identification of human procalcitonin
Tao FU ; Danhua SU ; Yuan LIU ; Qing LIU ; Liang WU ; Shengxia CHENG ; Xugan JIANG
Chinese Journal of Clinical Laboratory Science 2017;35(3):226-229
Objective To construct the prokaryotic expression vector of human procalcitonin(PCT) and obtain the highly purified recombinant PCT protein.Methods PCT primers were designed based on the sequence of PCT gene from NCBI,and PCT gene was amplified by PCR.Then,the recombinant vector PCT/pET-22b(+) was constructed and transferred into E.coli BL21 to induce the expression of recombinant PCT protein.Last,the recombinant PCT protein was purified by the nickel column affinity chromatography and identified by Western blot and the colloidal gold method,respectively.Results The results of agarose gel electrophoresis showed that the product of PCR amplification was about 350 bp.The homology comparison analysis revealed that the PCT gene fragment(348 bp) was successfully inserted into pTG19-T vector,and that no any base mutation was found.When the recombinant plasmid of PCT/pET-22b(+) was digested with BamH Ⅰ and HindⅢ,two pieces of about 350 bp and 5 500 bp were obtained.SDS-PAGE showed that the recombinant PCT protein with about 14 000 of Mr was existed in soluble form,and was easily obtained by the nickel column affinity chromatography.Moreover,western blot and the colloidal gold method demonstrated that the recombinant PCT protein was successfully expressed and contained histidine label(His-tag).Conclusion The recombinant vector PCT/pET-22b(+) is successfully constructed by the recombinant DNA technology,and the recombinant PCT fusion protein is successfully obtained by the nickel column affinity chromatography.
5.Cloning,fusion expression and identification of thioredoxin encoding gene from Toxoplasma gondii
Zigang ZHANG ; Xiaomei CHEN ; Danhua SU ; Yuan LIU ; Tao FU ; Jiamiao DUANMU ; Liang WU ; Xugan JIANG ; Shengxia CHEN ; Jianping CAO
Chinese Journal of Schistosomiasis Control 2016;28(3):289-292
Objective To clone and express the thioredoxin(Trx)from RH strain tachyzoites of Toxoplasma gondii,estab?lish the prokaryotic expression vector and purify the recombinant protein,then produce the polyclonal anti?Trx antibody in rab?bits. Methods Trx fragment was amplified by PCR and cloned into the pET?28a(+)vector,and the recombinant protein was in?duced with IPTG and purified by Ni?NTA affinity chromatography. The polyclonal antibody specificity was detected by Western blotting. Results The trx gene was amplified from T. gondii cDNA by PCR. The recombinant plasmid trx/pET?28a(+)was use?fully constructed,and the recombinant TRX protein was expressed and purified. The TRX polyclonal antibody was also ob?tained. The specific band of TRX was detected by Western blotting. Conclusion Western blotting can detect the specificity of polyclonal anti?Trx antibody,which will facilitate the biological functions of Trx.
6.Construction of comprehensive quality of evaluation index system about palliative care in general hospitals based on the structure-process-outcome quality model
Yan WU ; Limei SUN ; Yanbo JI ; Yujie XU ; Shengxia LIU ; Ruirui ZHAO
Chinese Journal of Practical Nursing 2022;38(5):360-364
Objective:To construct of comprehensive quality of evaluation index system about palliative care in general hospitals, so as to provide reference for promoting the scientific and standardized development of palliative care.Methods:Based on the structure-process-outcome quality model, literature research and Delphi method were used to determine the quality of palliative care evalution index system and index weight for general hospitals.Results:A total of 12 experts were consulted for two rounds,the rates of questionnaire retrieve were 12/15 and 12/12 respectively. The authoritative coefficients were 0.909 and 0.879, the Kendall′s W values were 0.27, 0.32 and 0.26 respectively with good coordination degree ( χ2=6.50, 106.62, 494.64, all P<0.05). Finally, the quality of palliative care indicator system in general hospitals was constructed, which included 3 first-level indicators, 30 second-level indicators and 157 third-level indicators. Conclusions:The establishment process of the construction of quality of palliative care indicator system in general hospitals was scientific and reasonable, focusing on the development characteristics of palliative care and can make significant contributions to improve the quality of palliative care.