2.Mechanism of liver and lung injury in septic mice
Chuan-Bo ZANG ; Rong-Qian WU ; Xu-Hua SONG ; Ying-Xin XU ; Rong LI ;
Academic Journal of Second Military Medical University 1985;0(06):-
Objective:To investigate the mechanism of liver and lung injury in mouse septic models.Methods:Twenty-four male Kunming mice were subjected to cecal ligation and puncture(CLP)or sham operation.The permeability of microvasculature,water contents,activities of myeloperoxidase(MPO)and the apoptosis of microvascular endothelial cells in lung microvasculature and liver sinus were examined 3 h and 12 h after operation.Results:Both the liver and lung showed a significant increase in microvessel permeability at 12 h in CLP group compared with sham operation group.MPO activity and water content in CLP group were obviously higher than those in the sham operation group.The apoptosis of lung microvascular endothelial cells at 12 h in CLP group(5.03?0.92)% was significantly higher than that of control group(3.48?1.21)%(P
3.Study of baicalin in inducing prostate cancer cell line DU145 apoptosis in vitro.
Zheng-Qin GU ; Ying-Hao SUN ; Chuan-Liang XU ; Yi LIU
China Journal of Chinese Materia Medica 2005;30(1):63-66
OBJECTIVEThe purpose of the present study was to investigate the in vitro effects of baicalin on induction of apoptosis in human prostate cancer cell line DU145.
METHODHuman prostate cancer cell line DU145 was treated with different concentration of baicalin in vitro. The apoptosis rate was determined by FACS analysis, cell cycle distribution was detected by flow cytometry, morphological changes and protein analysis were determined by means of electron microscope techniqueand immunohistochemical techniquerespectively.
RESULT50micromol x L(-1) and 125 micromol x L(-1) of baicalin dose-dependently induced apoptosis and inhibited the proliferation of prostate cancer cell DU145 in a dose and time-dependent manner. DNA flow cytometric analysis indicated that baicalin induced a arrest in G1 phase, showing a typical apoptosis peak. Electron microscopy detected a characteristic appearance of the apoptotic cells morphology. Immunohistochemical analysis revealed that induction of apoptosis by ways of inhibition of the bcl-2, loss of the Bax, and upregulation of Fas.
CONCLUSIONThe results indicate that baicalin may induce apoptosis and inhibit proliferation of prostate cancer cells, and has direct anti-tumor effects on human prostate cancer cells.
Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Flavonoids ; isolation & purification ; pharmacology ; G1 Phase ; Humans ; Male ; Plants, Medicinal ; chemistry ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Scutellaria ; chemistry ; bcl-2-Associated X Protein ; fas Receptor ; metabolism
4.Production of VEGF induced by GMCSF via ERK-NF-KB singling 'pathway in human fibroblasts during wound healing
Xiaoguang LI ; Min YAO ; Yong FANG ; Weirong YU ; Peng XU ; Ying WANG ; Chuan GU ; Yi WANG
Chinese Journal of Trauma 2011;27(8):731-736
ObjectiveTo observe production of vascular endothelial growth factor (VEGF) induced by granulocyte/macrophage colony-stimulating factor (GMCSF)via ERK nerve growth factor (NF)-κB singling pathway in human fibroblasts during wound healing and explore relating mechanism.MethodsHuman fibroblasts from the injured skin were used for this study and treated with GMCSF.RT-PCR was used for analyzing the protein and mRNA levels of VEGF and Western blotting was employed to determine the phosphorylation of ERK. The fibroblasts were pre-treated with ERK specific inhibitor PD98059 and further treated with GMCSF, then the fibroblasts and the supernatant were collected for detection of protein level of VEGF by means of Western blot. ERK signal pathway was inhibited to detect the activation of NF-κB by means of immunofluorescence staining. Furthermore, the nuclear and cytoplasmic extraction kit was used to separate the cytoplasm and nucleus and Western blot employed for observation of the NF-κB activation. ResultsThe mRNA level and protein level of VEGF were increased significantly with treatment with higher concentration of GMCSF in a dose-dependent manner. VEGF mRNA level was increased two hours after administration with GMCSF and reached peak at 4-6 hours. GMCSF could remarkably activate the ERK phosphorylation. Compared with GMCSF, the ERK specific inhibitor PD98059inhibited significantly the effect of GMCSF in inducing VEGF expression (P < 0.05). Western blot and immunofluorescence staining analyses showed that the activation of NF-ΚB was inhibited with reduced production of VEGF after GMCSF treatment.Conclusion GMCSF up-regulates production of VEGF through activating NF-κB via ERK signal pathway in the human fibroblasts.
5.Detection of apoptosis of Th1 and Th2 cells in C57BL/6 mice chronically infected with Schistosoma japonicum
Xinyu XU ; Jiaqing ZHAO ; Ying CHI ; Lei HE ; Xiaoyun WEN ; Chuan SU
Chinese Journal of Schistosomiasis Control 1989;0(01):-
Objective To observe the apoptosis of Th1 and Th2 cells in C57BL/6 mice chronically infected with Schistosoma japonicum.Methods The apoptotic Th1 and Th2 cells in spleen and lymph node from C57BL/6 mice infected with Schistosoma japonicum for 13 weeks were examined by three-color and indirect flow cytometery with staining surface molecule and intracellular cytokines.Results Compared with the normal mice,the proportion of apoptotic Th1 and Th2 cells of 13-week post-infection was significantly high,and the apoptotic Th1 cells increased more than apoptotic Th2 cells in the infected C57BL/6 mice,and the Th1 cells were more susceptible to apoptosis than Th2 cells.Conclusions Unequal susceptibility to apoptosis in Th1 and Th2 cells may be one of the reasons leading to Th2 polarization on mice chronically infected with Schistosoma japonicum,which provides the new proof of Th polarization.
6.Characteristics of proliferation and myotube cell formation of rat skeletal muscle satellite cells cultured in vitro
ke, XU ; zu-jun, FANG ; ying-chuan, LI ; jie, ZHENG ; qiang, DING
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(07):-
Objective To establish a method of isolation and purification of rat skeletal muscle satellite cells,and observe the characteristics of proliferation and myotube cell formation of skeletal muscle satellite cells cultured in vitro. MethodsPurified skeletal muscle satellite cells were obtained by improved two-step enzymatic digestion method and pre-plating technique.Immunohistochemical staining was employed to identify the skeletal muscle satellite cells cultured in vitro.The growth of skeletal muscle satellite cells cultured in vitro was examined by MTT assay.The differentiation of skeletal muscle satellite cells was observed by inverted microscopy. Results The skeletal muscle satellite cells with higher purity were obtained and confirmed by the high expression of desmin.When cultured in vitro,the latent phase of skeletal muscle satellite cells was the first to the second day,and the platform phase was the fifth to the sixth day.Myotube cells gradually formed when cell confluence was more than 60% to 70% or differential medium with lower fetal bovine serum was used.Conclusion The combination of improved two-step enzymatic digestion method and pre-plating technique serve as an easy and practical way to obtain skeletal muscle satellite cells with higher purity.Skeletal muscle satellite cells can form myotube cells with contraction characteristics without any special induction.
7.Effect of high glucose on expression of MIP-2 and MCP-1 in macrophages cultured in vitro
chuan, GU ; yong, FANG ; tao, NI ; wei-rong, YU ; peng, XU ; zhu-ying, GUO
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(11):-
0.05).The protein and mRNA expression of MIP-2 in high glucose group significantly increased after culture for 4 h,and guadually decreased then.The protein and mRNA expression of MCP-1 began to increase significantly after culture for 8 h,reached peak at 12 h,and slightly decreased after culture for 24 and 48 h. Conclusion High glucose promotes the protein and mRNA expression of MIP-2 and MCP-1 from mouse peritoneal macrophages cultured in vitro,which indicates that high glucose may delay the wound healing by increasing the expression of chemokines in diabetic mice.
8.Overexpression of β(1)-adrenoceptor can not protect rat cardiomyocytes from injury induced by isoprenaline.
Ying WANG ; Feng ZHOU ; Chuan-Ying XU ; Hong SUN
Acta Physiologica Sinica 2010;62(6):505-510
The purpose of this study was to investigate the effect of the overexpression of β(1)-adrenoceptor (β(1)-AR) on the contractile function and cell survival of rat cardiomyocytes injured by isoprenaline (ISO). The rat cardiomyocytes were isolated using the collagenase perfusion method and then transfected with β(1)-AR gene using adenoviruses vector. Four hours after the infection, the rat cardiomyocytes were treated with ISO for 24 h to imitate the high catecholamine levels of chronic heart failure. Western blot was performed to measure the protein expression of β(1)-AR. The percentages of rod cells were measured to test cell survival. Video-based edge-detection system was used to measure the contractile function of the cardiomyocytes. The results indicated that the expression of β(1)-AR in β(1)-AR-transfected cardiomyocytes was significantly increased compared with that in control group (P<0.01). Meanwhile, β(1)-AR transfection also increased β(1)-AR protein levels in ISO-injured cardiomyocytes. The cardiomyocyte survival was significantly decreased in ISO group compared with that in control group. β(1)-AR-transfection alone had no effect on cardiomyocyte survival in β(1)-AR group, but it further decreased cardiomyocyte survival in β(1)-AR+ISO group. Contractile amplitudes of ISO-injured cardiomyocytes were significantly decreased regardless of whether they were transfected with β(1)-AR or not, although β(1)-AR-transfected cardiomyocytes showed significantly increased contractile function compared with control group (P<0.05). These results suggest that the overexpression of β(1)-AR has no significant protective effect on rat cardiomyocytes injured by ISO.
Animals
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Cell Survival
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Cells, Cultured
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Female
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Heart Failure
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metabolism
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Isoproterenol
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pharmacology
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Male
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Myocardial Contraction
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drug effects
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Myocytes, Cardiac
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cytology
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metabolism
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pathology
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Rats
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Rats, Sprague-Dawley
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Receptors, Adrenergic, beta-1
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genetics
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metabolism
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Transfection
9.Detection of growth phenotype and blood biochemical parameters of wild type-derived chromosome 1 substitution mouse strain
Chuan GAO ; Wei XU ; Fuyi XU ; Yaoqi ZHANG ; Ying ZHAO ; Liya ZHAO ; Yuxun ZHOU ; Kai LI ; Junhua XIAO
Acta Laboratorium Animalis Scientia Sinica 2015;(3):233-238
Objective To analyze the growth phenotype and blood biochemical parameters of chromosome 1 substi-tution mouse strain(CSS1), and investigate their potential of QTL mapping .Methods Body weight, body length, tail length, organ weight of the CCS1 mice were measured at different days to create a growth curve while blood biochemical in -dexes were measured at about the 80th day.Results The CCS1 mice were different from C57BL/6 mice in several inde-xes.Compared with the C57BL/6 mice during different developmental stages , six strains including B6-Chr1KM mice were significantly different in body weight .There were five strains including B6-Chr1CM mice significantly different with C57BL/6 mice in body length, and all of the CSS1 mice were significantly different from C57BL/6 mice in tail length.Part of CCS1 mice were significantly different from C57BL/6 mice in the weight of liver, spleen, kidney and brain.The ALT of female B6-Chr1CM mice was significantly higher than that in the C 57BL/6 mice.The ALP of female B6-Chr1HZ mice was signifi-cantly higher than that in the male C57BL/6 and B6-Chr1KM mice, and was significantly lower than that in the C57BL/6 mice.The TB of male B6-Chr1CM, B6-Chr1SMX and B6-Chr1HZ mice was significantly higher than that of the C 57BL/6 mice.The TG of male B6-Chr1SMX mice and male B6-Chr1TW mice was significantly higher than that in the C 57BL/6 mice. Conclusions The phenotype of Chr1 CSS mice is quite different from commonly used inbred strain C 57BL/6 mice.CCS1 mice show great potential in QTL mapping for their characteristic growth phenotype and blood biochemical indexes .
10.Treatment of aneurysms rupture during endovascular embolization
Xu-Ying HE ; Chuan-Zhi DUAN ; Tie-Lin LI ; Qiu-Jing WANG ; Fang-Ming YIN ; Ru-Xiang XU ;
Journal of Interventional Radiology 1994;0(02):-
Objective To evaluate the treatment of aneurysms rupture during endovascular embolization.Methods Nine aneurysms ruptured during the embolization and were treated with endovascular embolization.The reasons of aneurysms rupture during embolization,the prevention and the first aid after aneurysms rupture were analysed.Results Seven patients recovered and 2 died.Conclusions The optimal treatment of aneurysms rupture during endovascular embolization is effective,(J Intervent Radiol,2007,16: 132-134)