1.Cloning, expression and purification of the human augmenter of liver regeneration (hALR) cDNA
Shimin CHEN ; Zailong CAI ; Jifang MAO ;
Academic Journal of Second Military Medical University 1985;0(06):-
Objective: To clone human augmenter of liver regeneration (hALR) cDNA, construct the recombinant expression vector, express and purify its product. Methods: The hALR cDNA was obtained by using RT PCR method with total RNA extracted from the fetal hepatic tissue. Then it was cloned into the pGEM T vector, and subcloned into expression vector pGEX 4T 3.After proved to be correct by sequencing, recombinant expression plasmid pGEX 4T 3(hALR) was transformed into E.coli BL21(DE3). The fusion protein GST hALR was produced by IPTG induction, isolated by affinity chromatography glutathione Sepharose 4B and cleaved by Thrombin. Results and Conclusion: Recombinant expression plasmid pGEX 4T 3(hALR) is constructed. The hALR is highly expressed in E.coli . The fusion protein in the plasma is resoluble. The procedure of purification and cleavage of fusion protein is easy and simple.
2.Preliminary study of brain function regional homogeneity in transient ischemic attack
Lan PENG ; Dongbai LIU ; Dinghua LIU ; Jianyu ZHANG ; Shimin CAI ; Xianping MENG
Chinese Journal of Postgraduates of Medicine 2012;35(25):7-9
ObjectiveTo investigate the brain function in the patients with transient ischemic attack (TIA) using index of regional homogeneity ( ReHo ).MethodsSixteen TIA patients ( TIA group ) and 16 age-matched normal controls(control group) underwent standard resting state functional MRI (fMRI)scan.The changes of the brain ReHo were studied by DPARSF analysis.ResultsCompared with that of control group,TIA group showed significantly decreased ReHo in the left cingulate gyrus (z =-3.72),left frontal gyrus (z =-3.02),right frontal gyrus (z =-3.23),right superior frontal gyrus (z =-3.75),right precuneus (z =-3.80),right inferior parietal lobule (z =-3.98 ),left inferior parietal lobule (z =-3.82),precentral gyrus ( z =- 3.85 ),right midfrontal gyrus ( z =-4.15 ),right midtemporal gyrus (z =- 3.43 ),and increased ReHo in the right hippocampus (z =3.37) and right cerebellum (z =3.55).Conclusion The rest-state brain function is abnormal in TIA interictal phase,and the increased ReHo in the hippocampus and cerebellum may reflect stress-induced brain protection of TIA and partial comoensatory response.
3.Mechanism of astragaloside IV promoting bone marrow EPCs mobilization in diabetic ulcer rats
Luyao ZHANG ; Shimin CAI ; Xi ZHANG ; Xiaoqin SONG ; Xiaoling ZOU ; Yuting XIAO ; Ying YANG ; Yang WEI ; Hongyu HUANG ; Wu XIONG
Journal of Chinese Physician 2024;26(3):376-381
Objective:To investigate the effect of astragaloside IV (AS-IV) regulating the signal axis of stromal cell-derived factor-1α (SDF-1α)/CXC chemokine receptor 4 (CXCR4) on the mobilization of bone marrow endothelial progenitor cells (EPCs) to peripheral blood in diabetes skin ulcer (DSU) rats.Methods:Twenty four SPF grade male Sprague Dawley (SD) rats were selected to make the model of type 2 diabetes rats by intraperitoneal injection of 30 mg/kg 1% (plastid ratio) streptozotocin, and then round full-thickness skin with a diameter of 2 cm was cut on both sides of the waist and back to make the skin ulcer model of diabetes rats. After that, they were randomly divided into AS-IV group (50 mg/kg AS-IV), blocker group (50 mg/kg AS-IV+ 5 mg/kg AMD3100) and model group. At the same time, a blank group ( n=8) was set up, The drug was administered via intraperitoneal injection, and the model group and blank group were treated with 0.9% NaCl of equal volume. On the 10th day, peripheral blood, femoral bone marrow, and wound neovascularization tissues of rats were collected. The number of EPCs in peripheral blood of each group of rats was measured by flow cytometry, and the protein expression of SDF-1α and CXCR4 in peripheral blood, femoral bone marrow, and wound neovascularization tissues of rats was detected by enzyme-linked immunosorbent assay (ELISA); At the same time, the wound healing rates of each group were tested. Results:On the 10th and 21st day after modeling, the wound healing rate of each group of rats was compared. The blank group healed the fastest, while the model group healed the slowest. The AS-IV group had better healing than the model group and the blocker group, and the difference was statistically significant (all P<0.05). On the 10th day after modeling, the positive rates of peripheral blood EPCs in the white group, AS-IV group, and blocker group were significantly higher than those in the model group (all P<0.05), while the positive rates of peripheral blood EPCs in the blocker group were significantly lower than those in the AS-IV group (all P<0.05). On the 10th day after modeling, the protein expression of SDF-1α and CXCR4 in the wound, serum, and bone marrow of the model group was the lowest, while the protein expression in the blank group was the highest (all P<0.05). The protein expression of SDF-1α and CXCR4 in the wound, serum, bone marrow of the AS-IV group was significantly higher than that of the blocker group and model group, and the difference was statistically significant (all P<0.05). Conclusions:Astragaloside IV can promote the mobilization and migration of endothelial progenitor cells from bone marrow to peripheral blood in diabetes ulcer rats by regulating SDF-1α/CXCR4 signal axis, and can participate in angiogenesis of diabetes ulcer wounds as seed cells to promote the healing of diabetes skin ulcers.
4.Application of Non-pharmacotherapy Treating Cervical Radiculopathy in Real-world Study
Xu WEI ; Yun GAO ; Qi XIE ; Liguo ZHU ; Xingping ZHANG ; Jinghua GAO ; Yong ZHAO ; Jinxue LI ; Shimin ZHANG ; Zhenyu ZHANG ; Jingyi CAI ; Yongli DONG ; Hao WANG ; Bin TANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2017;19(12):1960-1965
This study was aimed to analyze the application of non-pharmacotherapy in treating cervical radiculopathy (CR) in real-world,and to provide clinical reference for CR non-pharmacotherapy.The clinical real-world data of CR was extracted by using information sharing system of traditional Chinese medicine (TCM) and clinical research.Six hundred and twenty-eight inpatients and outpatients with CR were enrolled from December 2012 to July 2014 in the information system database of Wangjing Hospital.Basic characteristics of the non-pharmacotherapy groups were analyzed by statistical description method.The node degree and mutual information value were recorded for non-pharmacotherapy application of all patients by using liquorice software.Complex network diagrams were generated.The results showed that 47% of CR patients received non-pharmacotherapy (294/628),including 67 males and 227 females.The average age of patients was 49 years old,and the prevalence of the disease was the highest from 45 to 65 years old.In all patients,the usage of manipulation and cervical traction was higher,and the combination of manipulation and acupuncture was the most.Within outpatients,the proportion of cervical traction was higher,and the combination of manipulation and acupuncture was the most frequently.Within inpatients,the proportion of manipulation and cervical traction was higher,and the combination of comprehensive physical therapy and exercise therapy was the most frequently.It was concluded that non-pharmacotherapy has been commonly used in clinical treatment of CR.Cervical traction and manipulation was the widest applications.The combination treatment was in wide application.Future studies should increase the sample size of CR patients from different regions,and enhance gradually the level of evidence of clinical research for non-pharmacotherapy treating CR.