2.Mechanism of MCP-1-JAK2/STAT3 signal transduction in spinal cord dorsal horn contributing to rats with type 2 diabetic neuropathic pain
Chao YUAN ; Jinchuan GUAN ; Hong CHAO ; Jun LI
Chinese Journal of Pathophysiology 2015;(8):1467-1471
AIM:ToexplorewhetherMCP-1-JAK2/STAT3signaltransductioninthespinaldorsalhornin-volves the formation and development of rat type 2 diabetic neuropathic pain (DNP).METHODS: The male Sprague-Dawley rats were fed with a high-fat and fructose diet for 8 weeks,and then received a single intraperitoneal streptozocin in-jection to prepare the type 2 DNP model.The type 2 DNP rats were randomly divided into 4 groups (n=16):DNP group, MCP-1 neutralizing (DM) group, DNP+AG490 (DA) group and solvent control (SC) group.A catheter of PE-10 was placed into the subarachnoid space of the rats in groups DM , DA and SC.After 3 d, the rats in DM,DA and SC groups were injected with MCP-1 inhibitor 10μL at 0.1 mg/L, AG490 10μL at 1 mmol/L and DMSO 10μL at 3.5%once a day for 14 days, respectively.Another 16 normal rats were selected as control (C) group and were fed with common forage. The mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were measured at 1, 3, 7 and 14 d after subarachnoid injection .The lumbar segments 4-6 of the spinal cord were removed at the same time for determination of the expressions of p-JAK2 and p-STAT3 by Western blot .RESULTS:Compared with C group , MWT was significantly de-creased, TWL was shortened and the expression of p-JAK2 and p-STAT3 in the spinal dorsal horn was up-regulated at 1, 3, 7 and 14 d in DNP and SC groups (P<0.05).Compared with DNP group, MWT was significantly increased, TWL was prolonged and the expression of p-JAK2 and p-STAT3 in spinal dorsal horn was down-regulated at 1, 3, 7 and 14 d in DM and DA groups (P<0.05).No significant difference in the MWT, TWL and expression of p-JAK2 and p-STAT3 between DNP group and SC group was observed (P>0.05).CONCLUSION:The MCP-1-JAK2/STAT3 signal transduction in the spinal dorsal horn involves the formation and development of DNP in rats .
4.Indication of video-assisted thoracoscopic decortication in treatment of tuberculous empyema
Jun WANG ; Chao CUI ; Jun ZHANG ; Miao LI ; Lei XU
China Journal of Endoscopy 2016;22(7):98-101
Objective To evaluate the efficacy of decortication by video-assisted thoracic surgery (VATS) in pa﹣tients with tuberculous empyema, and discuss its indications. Methods 60 patients with tuberculous empyema who underwent decortication by VATS for surgical management from December 2010 to December 2015 were included. Under a thoracoscope, we cleaned up the pus, separated adhesions, scraped granulation tissues and caseous necrosis on the inner wall of the abscess cavity, and stripped the thickened fiberboard of the parietal and visceral pleurae. Af﹣ter the procedure, sufficient drainage and antituberculosis therapy were carried out. Results All the patients in this group were operated successfully. All the patients were cured without perioperative death and complications. No re﹣currence of empyema was observed at the follow-up examination from 2 months to 5 years, and suffered pulmonary reexpansions were better. Conclusions The decortication by VATS for tuberculous empyema is safe, effective, mini﹣mally invasive. The imaging manifestations of pleural thickening in 1 cm, no obvious calcification, no serious lesions in the lungs are the indications for the operation.
5.An exprimental study on AZD5363 suppressing cholangiocarcinoma cells
Chao WU ; Yuncheng ZHANG ; Zhe LI ; Guangbing LI ; Jun LIU
Chinese Journal of General Surgery 2016;31(1):55-58
Objective To investigate the effect of Akt pathway inhibitor AZD5363 on cell proliferation and invasion of QBC939 and RBE cholangiocarcinoma cells and the mechanism.Methods Western blotting was used to detect Akt and downstream protein and mTOR protein expression in two cancer cell lines after process by AZD5363.Inhibition rate and cytotoxicity was tested by CCK-8 assay,and Transwell assay was used to evaluate the invasive ability of cancer cells.Results QBC9393 cell exposed to AZD5363 LD50 drug concentration (24 ±9) was significantly different compared with control group (t =4.47,P < 0.05),RBE cells LD50 drug concentration (21 ± 8) was significantly different compared with control (t =4.41,P < 0.05).Tumor invasion capacity of QBC939 in drug concentrations of 20 μmol/L (63 ± 12) and 0 μmoL/L (271 ± 27),the difference was statistically significant.RBE exposed AZD5363 upon drug concentrations of 20 μmol/L (58 ± 23) and 0 μmol/L (235 ± 21),the difference was statistically significant.AZD5363 promotes phosphorylation of mTOR in QBC939.Conclusions AZD5363 inhibits the proliferation and migration,inhibiting the phosphorylation of Akt and its downstream molecules.AZD5363 promotes phosphorylation of mTOR in QBC939.
6.Effect of Flos Daturae Alkaloids on TGF-beta1-induced Epithelial-Mesenchymal Transition of Human Pulmonary Adenocarcinoma A549 Cells.
Zhen WANG ; Jun-chao YANG ; Xiao-li TAN ; Xin LV
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(11):1340-1344
OBJECTIVETo study the effect of Flos Daturae alkaloids (FDA) on TGF-beta1-1uuuu;U epithelial-mesenchymal transition (EMT) of human pulmonary adenocarcinoma A549 cells.
METHODSA549 cells in vitro cultured were randomly divided into 5 groups, i.e., the blank control group, the TGF-beta1 group, the low dose FDA group, the medium dose FDA group, and the high dose FDA group. The morphologies of A549 cells were observed. Expressions of cytokeratin (CK)-19 and alpha-smooth muscle actin (alpha-SMA) were detected by Western blot and real-time PCR at 24, 48, and 72 h, respectively.
RESULTSA549 cells in the TGF-beta1, group turned from cobblestone to spindle shape gradually. Those in low, medium and high dose FDA groups showed similar shapes to those of the TGF-beta1 group. There was no statistical difference in the morphology of A549 cells among the 3 dose FDA groups (P > 0.05). Western blot showed that, when compared with the blank control group, the expression of CK-19 was down-regulated, but the expression of alpha-SMA was up-regulated in the TGF-beta1 group (P < 0.01). Compared with the TGF-beta1, group, the expression of CK-19 was up-regulated, but the expression of alpha-SMA was suppressed in low, medium and high dose FDA groups (P < 0.01). The CK-19 expression obviously increased, but the alpha-SMA expression was suppressed in high dose FDA group at 72 h (P < 0.01). Real-time PCR results showed, as compared with the TGF-beta1 group, the mRNA expression of CK-19 was increased, but the mRNA expression of alpha-SMA was reduced in low, medium and high dose FDA groups (P < 0.01).
CONCLUSIONSFDA had no effect on EMT morphological changes of TGF-beta1 induced A549 cells. FDA could reverse characteristic markers of A549 cells during EMT to some extent, such as expressions of CK-19 and alpha-SMA. The expression of CK-19 (as the epithelium marker) increased and the expression of alpha-SMA (as the mesenchymal marker) was reduced. Besides, they were most obviously seen in the high dose FDA group at 72 h in a dose- and time-dependent manner.
Actins ; Adenocarcinoma ; Alkaloids ; pharmacology ; Cell Line, Tumor ; Datura ; Drugs, Chinese Herbal ; pharmacology ; Epithelial Cells ; Epithelial-Mesenchymal Transition ; Epithelium ; Humans ; Transforming Growth Factor beta1 ; metabolism
8.Effect of curcumin on tumor necrosis factor-alpha-induced expression and release of monocyte chemoattractant protein-1 in rat astrocytes
Ruixian WANG ; Chao YUAN ; Lulu SHEN ; Jun LI ; Hong CAO
Chinese Journal of Anesthesiology 2013;33(8):916-919
Objective To investigate the effect of curcumin on tumor necrosis factor-alpha (TNF-α)-in-duced expression and release of monocyte chemoattractant protein-1 (MCP-1) in rat astrocytes.Methods The primary astrocytes were prepared from the cerebral cortex of 5 neonatal Sprague-Dawley rats and cultured.The cultured cells were identified by immunofluorescence staining with glial fibrillary acid protein.The cells were then divided into 6 groups (n =15 each):control group (group C),TNF-α group (group T),TNF-α+ different concentrations of curcumin groups (Cur5,Cur10 and Cur20 groups),and TNF-α+ solvent control group (D group).TNF-α with the final concentration of 20 ng/ml was added and the cells were incubated for 2h in T group.In Cur5,Cur10 and Cur20 groups,curcumin with the final concentrations of 5,10 and 20μmol/L was added,respectively,the cells were incubated for 24h and then the culture medium was abandoned,TNF-α with the final concentration of 20 ng/ml was added and the cells were then incubated for another 2h.In group D,dimethyl sulfoxide with the final concentration of 1 μl/ml was added,the cells were then incubated for 24h,then TNF-α with the final concentration of 20 ng/ml was added and the cells were incubated for another 2h.After treatment in each group,the expression of MCP-1 was determined by immunohistochemistry and the release of MCP-1 was determined by ELISA.Results Compared with group C,the expression and release of MCP-1 was significantly increased in the other five groups (P < 0.05).Compared with group T,the expression and release of MCP-1 was significantly decreased in group Cur20 (P < 0.05),and no significant changes in the expression and release of MCP-1 were found in Cur5,Cur10 and D groups (P > 0.05).Compared with group Cur5,the expression and release of MCP1 was significantly decreased in group Cur20 (P < 0.05),and no significant change in the expression and release of MCP-1 was found in group Cur10 (P > 0.05).Conclusion Curcumin can inhibit TNF-α-induced expression and release of MCP-1 in rat astrocytes and the effect is dose-related and may be one of the mechanisms of curcumin-induced reduction of neurophathic pain.
9.Peripheral blood stem cell transplantation in the treatment of dilated cardiomyopathy: A 3-month follow-up in 42 cases
Tong ZHANG ; Chao LIU ; Jun YANG ; Xiang ZHANG ; Wen LI
Chinese Journal of Tissue Engineering Research 2007;0(16):-
AIM: Recent studies showed that stem cells could replace injured cardiomyocyte and increase the number of functional cardiomyocytes. Researching the pertinent literature in China Journal Full-text Database (CJFD) published between 2005 and 2008 indicated that the researches on stem cell transplantation in the treatment of primary dilated cardiomyopathy were few. This study investigated the therapeutic efficacy and security of peripheral blood stem cell transplantation in coronary artery in treatment of dilated cardiomyopathy and its effects on left ventricular function. METHODS: Forty-two patients with dilated cardiomyopathy between December 2006 and September 2007 were enrolled at the Department of Cardiology of First People's Hospital of Yunnan, including twenty-eight males and fourteen females, averagely aged (56?3) years. Inclusive criteria: patients with less than 65 years, left ventricular enlargement, and left ventricular ejection fraction (LVEF) ≤ 45%, and without coronary artery disease after coronary arteriongraphy. Informed consents were obtained from patients. Patients were divided into stem cell transplantation group (n=15) and control group (n=27) on the basis of whether being treated by stem cell transplantation. Patients in the stem cell transplantation group were consecutively administered granulocyte colony-stimulating factor (G-CSF) by hypodermic injection to stimulate bone marrow stem cells themselves based on conventional treatment for five days. Peripheral blood stem cell suspension was disassociated on the 6th day, and the collected suspension was injected into left anterior descending branch over the wire saccule tube for autologous peripheral blood stem cell transplantation. Patients in the control group were administered by conventional treatment of dilated cardiomyopathy. Security and adverse reaction were observed during the mobilization, collection and returning injection of peripheral blood stem cell by coronary artery. Morphous, cardiac function and motion index of left ventricle wall were evaluated using ultrasoundcardiogram before and 3 months after transplantation. Survival rate and incidence rate of heart incidents were compared. RESULTS: Three months after stem cell transplantation in coronary artery, there were a significant decrease in cardiac end-systolic volume (ESV), cardiac end-diastolic volume (EDV) and motion index of left ventricle wall, but a significant increase in LVEF(P
10.In vitro study of transfection of mesenchymal stem cells with adenoviral vector overexpressing human uPA
jun-jie, HU ; chao, SUN ; ling, LAN ; ding-guo, LI
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(04):-
Objective To identify the isolated rat bone marrow derived mesenchymal stem cells(MSCs),and evaluate the efficiency of adenoviral vector expressing human urokinase type plasminogen activator(uPA) in transfection of rat MSCs and its effect on proliferation of MSCs. Methods MSCs were isolated and purified by pasted wall purification,and were identified by immunicytochemistry.The transfection efficiency of uPA was detected by fluorescent microscopy,the expression of uPA in MSCs was detected by Western blotting,and the proliferation of MSCs was evaluated by MTT. Results The harvested MSCs exhibited the typical appearance of MSCs,and it was revealed by immunohistochemistry that the expression of MSCs markers CD29 and CD90 was positive,while that of CD34 and CD45 was negative.A tendency of increase in expression of green fluorescent protein(GFP) was observed with increase of multiplicity of infection(MOI).After transfection with AduPA for 72 h,the transfection efficiency reached(94.0?1.5)% at MOI of 80,and positive GFP cells could still be observed even after 7 d.The transfected uPA had no effect on the proliferation of MSCs. Conclusion MSCs are favourable genetic vectors to express uPA,and can be used for treatment of liver fibrosis.