1.Transforming growth factor-beta 1 at different concentrations induces the differentiation of bone marrow derived mesenchymal stem cell into smooth muscle-like cell
Yi CHEN ; Wenwei CAI ; Jing SHENG
Chinese Journal of Tissue Engineering Research 2009;13(45):8833-8837
BACKGROUND:Bone marrow derived mesenchymal stem cells (BMSCs) will be homing to the lesions after balloon injury in the inflammatory reaction process.However,the molecular mechanism of transforming growth factor-β1 (TGF-β1) to promote BMSC into smooth muscle-like cell remains unclear.OBJECTIVE:To investigate the BMSC differentiation rate under different TGF-β1 levels after acute vascular injury.DESIGN,TIME AND SETTING:A randomized controlled animal experiment and in vitro induced cell observation were performed at the SPF Laboratory Animal Center and Laboratory of Tissue Engineering,Ninth People's Hospital of Shanghai Jiao Tong University Medical School between January 2008 and May 2009.MATERIALS:A total of 24 male SD rats were randomly divided into normal control and model groups with 12 animals in each group.In addition,6 4-weak-old male SD rats were used to prepare BMSCs.METHODS:Model of acute carotid artery injury was established in model group.Serum of normal control group and model group after 1,3,7 days of injury to detect TGF-β1 level after the vascular injury by ABC-ELISA.The BMSCs after one passage were cultured at a density of (1.0-3.0)×10~5 cells/100 mm in culture dish,and divided into two groups:in routine culture group,cells were cultured in high-glucose DMEM containing 20% fetal bovine serum;in TGF-β1 group,cells were induced by different concentrations of TGF-β1 (1,3,5 and 10 μg/L) based on routine culture for 1 week.MAIN OUTCOME MEASURES:Serum TGF-β1 level after the vascular injury;,cell morphological changes by inverted phase contrast microscopy;smooth muscle α-actin expression.RESULTS:The normal serum TGF-β1 level in rat was low,but increased rapidly after 1 day of injury,reached its peak at 3 days,and declined gradually but did not return to the basic level till day 7.After 1 week of induction,BMSCs were confluent,forming peak valley appearance.Immunocytochemistry showed that compared with routine culture,the rate differentiation of smooth muscle-like cell was significantly increased in cells stimulated by TGF-β1,especially 5 and 10 μg/L TGF-β1 (P < 0.01).Real-time quantitative PCR results were similar to immunocytochemistry.CONCLUSION:Serum TGF-β1 level increased after acute vascular injury and peaked at day 3.In vitro,a similar concentration of TGF-β1 could induce cultured BMSC to differentiate into smooth muscle-like cells.
2.Analysis of the relationship between the psychological factors and effect of reduce blood pressure in aged hypertension
Jian ZHU ; Wenwei CAI ; Juxian YANG
Chinese Journal of Behavioral Medicine and Brain Science 2001;10(2):109-110
Objective To analyse the relationship between the psychological factors and effect of reduce blood pressure. To study the influence of the psychological factors in therapeutic effect of reduce blood pressure. Methods While using reduce blood pressure drug, behavioral intervention and biofeedback technique were used to analyse the relationship between the psychological factors and the effect of reduce blood pressure. Results The change values were positive correlation with the psychological factors and the reduce blood pressure. The change data of Te, E, Hy significantly affected the change data of blood pressure. Conclusion Behavioral intervention and biofeedback technique could increase the effect of reduce blood pressure drug in aged hypertension, improve their psychological quality of life.
3.Changes of neurobehavior and expression of tyrosine hydroxylase messenger RNA in mesencephalon of rat parkinsonism model
Wenwei LI ; Dingfang CAI ; Xiaohong CHEN
Chinese Journal of Geriatrics 2000;0(06):-
0 05). The forelimb functional tests demonstrated that, the number of food pellets eaten by PD rats with the contralateral side of 6 OHDA lesion (17 71?4 82) was significantly reduced compared with the controls (34 43?3 55)( P
4.Effect of peroxisome proliferators-activated receptor gamma agonist on phenotypic transition of adventitia fibroblasts induced by transforming growth factor beta 1 in vitro
Shaojun MA ; Wenwei CAI ; Jing SHENG
Chinese Journal of Tissue Engineering Research 2007;0(02):-
BACKGROUND: Previous studies demonstrated that adventitia fibroblasts exhibit important role in the hyperplasia of newly born endomembrane after blood vessel injury. However, the mechanism regarding phenotypic transition of adventitia fibroblasts in vitro remains unclear. OBJECTIVE: To observe effect of peroxisome proliferators-activated receptor ?agonist (PPAR ?) on phenotypic transition of adventitia fibroblasts induced by transforming growth factor ?1 (TGF-?1) in vitro, as well as the expression of collagen protein Ⅰ. DESIGN, TIME AND SETTING: A randomized grouping control experiment was performed at the Tissue Engineering Laboratory of Ninth Affiliated Hospital of Shanghai Jiao Tong Medical College between July 2007 and October 2008. MATERIALS: The aorta thoracica's adventitial fibroblasts were cultured from Sprague Dawley rats, with 2-month-old, weighing 120 g. The TGF-?1 was supplied by American GenWay Biotech Company. The rosiglitazone power was purchased from Beijing Comens Chemical Co., Ltd. METHODS: The experiment was divided into 3 groups: In the TGF-?1 group, fibroblasts was induced with TGF-?1 with different concentrations (3, 5, 10, 15 ?g/L) for 48 hours; in the rosiglitazone group, fibroblasts was induced with rosiglitazone (5, 10, 30, 50 ?mol/L) for 45 minutes followed by co-culture with 10 ?g/L TGF-?1 for 48 hours. There was no intervention measure in the control group. MAIN OUTCOME MEASURES: The phenotypic transition of fibroblasts was measured by ?-smooth muscle actin, the expression of smooth muscle ? 2-actin, as well as collagen type Ⅰ was determined by protein immunoblotting. RESULTS: Compared with the control group, 5 ?g/L TGF-?1 could significantly stimulate the changes of phenotypic transition of fibroblasts as well as the expression of smooth muscle ?2-actin and collagen type Ⅰ, which reach a peak level when treated with 10 ?g/L TGF-?1 (P 0.05). Compared with control group, 30 ?mol/L rosiglitazone could significantly inhibit changes of phenotypic transition of fibroblasts as well as the expression of smooth muscle ? 2-actin and collagen type Ⅰ(P
5.The effects and the mechanisms of exogenous fatty acids on colorectal cancer cells
Wenwei CAI ; Tianfang HUA ; Jun CAO ; Qing GU ; Dun SHI ;
Parenteral & Enteral Nutrition 1997;0(01):-
Objectives: To observe the effects and the mechanisms of exogenous fatty acids on colorectal cancer cells(LoVo). Methods:The effect of exogenous fatty acids on LoVo cells was determined by 5 diphenyl tetrazolium bromide(MTT) assay.The content of MDA was examined to evaluate the extent of lipid peroxidation. Results:The growth of LoVo cells was inhibited by polyunsaturated fatty acids(PUFAs).The inhibitory effect of saturated fatty acids and monounsaturated fatty acids on LoVo cells was not showed.None of the fatty acids was found to fibroblast cells(HLF).PUFAs caused the significant rising of intracellular MDA content. Conclusions:PUFAs could inhibit the growth of colorectal cancer cells.The strengthening of lipid peroxidation may be one of the mechanisms.
6.The change in metabolism of fatty acids in breast cancer cells and its significance
Wenwei CAI ; Tianfang HUA ; Jun CAO ; Qing GU ; Dun SHI ;
Parenteral & Enteral Nutrition 1997;0(02):-
Objectives:To observe the changes in metabolism of fatty acids in breast cancer cells and the effects of inhibiting fatty acid synthase(FAS) on the growth of breast cancer cells. Methods:By RT PCR,the expression of FAS mRNA in breast cancer cell line was examined.The growth inhibition of breast cancer cells by specific FAS inhibitor, cerulenin, was determined by MTT assay.Flow cytometric analysis was used to study the changes of cell cycle. Results:FAS mRNA expression in MCF 7 cells was high.We found that cerulenin caused dose and time dependent inhibition of growth of MCF 7 cells.The growth inhibition of MCF 7 after 48 hours exposure of cerulenin at 2.5,5,10 and 20 mg/L was ( 43.47 ?4.58)%,(62.92?2.68)%,(81.93?0.91)% and (67.7?12.27)%( P
7.Protecting effect of Cistanche extracts on MPP+-induced injury of the Parkinson's disease cell model
Hu WANG ; Wenwei LI ; Dingfang CAI ; Ru YANG
Journal of Integrative Medicine 2007;5(4):407-11
OBJECTIVE: To investigate the potential protective role of Cistanche extracts on 1-methyl-4-phenylpyridinium ion (MPP(+))-induced Parkinson's disease (PD) cellular model, and to find out whether this effect is achieved through the regulation of growth arrest- and DNA damage-induced gene 153 (GADD153). METHODS: MPP(+))-induced cellular injury and the protective effect of Cistanche extracts on the SH-SY5Y cell line viability treated by MPP(+)) were investigated by using methyl thiazolyl tetrazolium (MTT) assay. The mRNA of GADD153 in SH-SY5Y cell line treated by MPP(+)) and Cistanche extracts were evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR). The protein level of GADD153 in SH-SY5Y was assessed by Western blotting. RESULTS: Cistanche extracts (100 mug/ml) increased the cell viability (P<0.01). And the mRNA of GADD153 in the Cistanche extracts pretreatment group was much less than that in the MPP(+)) group (P<0.01). The result of Western blotting showed that GADD153 had a lower level in the Cistanche extracts pretreatment group, compared with MPP(+)) group, especially in the 100 microg/ml group (P<0.01). CONCLUSION: Cistanche extracts pretreatment has a protective effect on the MPP(+))-treated SH-SY5Y cell line, and its down-regulation of GADD153 may contribute to the effect.
8.An in vitro study of thrombomodulin-protein C-EPCR system coagulation function in hemorrahgic shock rats
Ye SHEN ; Wenwei CAI ; Shuanghua WU ; Meiqi ZHANG ; Jingye PAN
Chinese Journal of Emergency Medicine 2012;(12):1354-1357
Objective To investigate the coagulant function of Thrombomodulin-Protein C-EPCR System in rats with hemorrhagic shock in virto study.Methods In this study,30 rats were divided into three groups,with each group 10 rats:control group,the sham shock group,hemorrhagic shock group.Hemorrhagic shock model was subjected to computer-controlled arterial hemorrhage to 40 mm Hg for 60 min The rat blood serum samples were obtained from rats in sham shock group and shock group after maintaining 3 hours,and fetal bovine serum (FBS) was used as control group.Then medium contained fore-mentioned serums cultured with SV40-transformed aortic rat endothelial cells for 6 hours,12 hours and 18hours.The messenger RNA (mRNA) expression of thrombmodulin (TM),Endothelial cell protein C receptor in the SV40-transformed aortic rat endothelial cells which were treated with serums were detected by reverse-transcription polymerase chain reaction.Results Compared to control group,SVARECs were incubated for 6hrs with shock serum,the mRNA expression of TF,TM and t-PA was significantly higher,and continued elevating at the time of 18 hours (P < 0.01).Conclusions Serums from hemorrhagic shock rats upregulate TMmRNA EPCRmRNA expression on the endothelial cells surface.It is suggested that Thrombomodulin-Protein C-EPCR System pay a key role of anticoagulation,the protection of endothelial function.
9.Early diagnostic value of soluble triggering receptor expressed on myeloid cells1 in patients with ventilator associated pneumonia
Xianghong YANG ; Renhua SUN ; Minhua CHEN ; Wenwei CAI
Chinese Journal of Emergency Medicine 2014;23(2):200-203
Objective To evaluate the early diagnostic values of soluble triggering receptor expressed on myeloid cell-1 (sTREM-1) in patients with ventilator associated pneumonia (VAP).Methods A total of 112 clinical suspicion VAP ventilator-associated pneumonia (VAP) patients accepted from January 2008 through December 2010 were enrolled for prospective and observational study.Two independent intensivists without aware of the results of the sTREM-1 assay separately made diagnosis of VAP present or absent depending on the clinical symptoms and results of microbial culture.Patients were categorized into two groups:VAP group (n =74) and non-VAP group (n =38).The levels of sTREM-1 in broncho-alveolar lavage fluid (BALF) collected with Gibot method in unemployment of bronchoscope and in serum were measured by enzyme-linked immunosorbent assay (ELISA) on the first day of suspected diagnosis.Comparison of sTREM-1 level between BALF and serum was made by t-test and Receiver Operating Characteristic (ROC) curve.Results A total of 112 clinical suspicion VAP patients admitted from January 2008 through December 2010 were enrolled for prospective and observational study.Two independent intensivists without aware of the results of the sTREM-1 assay made diagnosis of VAP present or absent depending on the clinical symptoms and results of microbial culture.Patients were categorized into two groups:VAP group (n =74) and non-VAP group (n =38).The levels of sTREM-1 in broncho-alveolar lavage fluid (BALF) collected with Gibot method in unemployment of bronchoscope and in serum were measured by enzyme-linked immunosorbent assay (ELISA) on the first day of suspected diagnosis.Comparison of sTREM-1 level between BALF and serum was made by t-test and Receiver Operating Characteristic (ROC) curve.Conlclusions In suspected VAP patients,measurement of sTREM-1 levels in BALF and in serum could help identify VAP in early stage.
10.Treatment of hair apposition technique with tissue glue on scalp lacerations
Wenwei CAI ; Yueliang ZHENG ; Xin CHEN ; Haifei HE ; Jianfeng TU
Chinese Journal of Emergency Medicine 2008;17(6):638-641
Objective To treat scalp lacerations by using the hair apposition technique (HAT) and to compare the HAT with standard suturing in a controlled prospective trial. Method Fifty patients with scalp lacerations were treated either by HAT or by standard suturing. Two groups were evaluated in consumed times for operation, pain sores, and complications. Results There were 30 HAT patients and 20 patients treated with suturing. The took shorter operation time consumed[(3.33 vs. (6.05 t = 4.85.P < 0.01], and HAT produced significantly lower pain score [(1.73vs. (3.20t = 4.01,P < 0.01]. There was a trend that more and more patients were willing to have HAT performed. Conclusions The advantages of HAT include a shorter time consumed for operation, less pain, satisfactory wound healing, and high acceptance by patients. HAT is acceptable for treating scalp lacerations in emergency room.