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Objective To investigate the effects of dehydrocostuslactone(Dehy) on human hepatic stellate LX-2 cells proliferation,apoptosis and the possible related mechanisms.Methods After treating the LX-2 cells by different concentrations of Dehy,the MTT assay and flow cytometry were used to assess the influence of Dehy on cell proliferation and cycle distribution of LX-2 cells.The Hoechst 33342 staining and AV-PI dual staining were used to detect the effect of Dehy on cell apoptosis in LX-2 cells.The apoptosis related proteins expression was detected by Western blot.Results After different concentrations of Dehy acting for 48 h could significantly inhibit the proliferation of LX-2 cells,blocked the cellular cycle at stage S and G2/M,meanwhile induced LX-2 cells apoptosis in the concentration dependent manner;Western blot results showed that Dehy could promote the up-regulation of P27 and bax expression and down-regulation of Bcl-2.Conclusion Dehy induces human liver LX-2 cells apoptosis and cell cycle arrest by adjusting the expression of Bax,Bcl-2 and P27.
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Objective To investigate the effects of dehydrocostuslactone(Dehy) on human hepatic stellate LX-2 cells proliferation,apoptosis and the possible related mechanisms.Methods After treating the LX-2 cells by different concentrations of Dehy,the MTT assay and flow cytometry were used to assess the influence of Dehy on cell proliferation and cycle distribution of LX-2 cells.The Hoechst 33342 staining and AV-PI dual staining were used to detect the effect of Dehy on cell apoptosis in LX-2 cells.The apoptosis related proteins expression was detected by Western blot.Results After different concentrations of Dehy acting for 48 h could significantly inhibit the proliferation of LX-2 cells,blocked the cellular cycle at stage S and G2/M,meanwhile induced LX-2 cells apoptosis in the concentration dependent manner;Western blot results showed that Dehy could promote the up-regulation of P27 and bax expression and down-regulation of Bcl-2.Conclusion Dehy induces human liver LX-2 cells apoptosis and cell cycle arrest by adjusting the expression of Bax,Bcl-2 and P27.
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Objective The aim of this study was to establish a rat model of blood hypercoagulable state by intra?venous injection of thrombin and to provide a model for researches on hypercoagulable state. Methods Rats were divided into six groups and were injected with normal saline and 2?5, 5, 10, 20, 40 U/kg thrombin solution through the femoral vein, respectively. Then, blood was drawn to test the activated partial thromboplastin time (APTT), prothrombin time ( PT) and fibrinogen ( FIB) , and to observe the death rate of rats in these groups to verify the optimal dosage. On this ba?sis, rats were injected thrombin of the best dose through the femoral vein, and blood samples were collected at 0, 10, 30, 60, 120, 180, 300 (s) to test APTT and PT and FIB for determining the best time for blood sampling. At last, the rats were divided into control group and thrombin group to inject normal saline or thrombin solution in the best dose via the fem?oral vein, and blood was taken at the best time to test APTT, PT, FIB and whole blood viscosity. Results APTT and PT values of the 10 U/kg thrombin group were the shortest, and FIB value of this group was the highest among these groups. APTT and PT values of blood sample collected at about 60 s after thrombin injection were the shortest, and FIB value was the highest. Compared with the control group, PT and APTT values of the thrombin group were shorter (P<0?05), and blood viscosity and FIB were higher ( P<0?05 ) . Conclusions Injecting thrombin solution into the femoral vein can be used to establish a rat model of hypercoagulable state. The best dose of thrombin solution is 10 U/kg in a concentration of 2 U/mL. The best time to collect blood sample is 60 s.
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Objective To explore the reversion effect of Gemcitabine-resistance A549 cell (A549/Gem) by silencing CXCR4.Methods A549 cell was induced by continuous stepwise exposure to Gemcitabine in order to obtain Gemcitabine-resistance A549 cell ( A549/Gem) in vitro.The CXCR4 expressions level of A549 and A549/Gem were detected by Quantitative RT-PCR ( RT-qPCR) and Western blot analyses.The CXCR4 shRNA vector was transfected into the A549/Gem cell by targeting silence CXCR4.Furthermore, MTT assay was used to explore the IC50 and RI in A549, A549/Gem and A549/Gem-CXCR4 cells.Moreover, Western blot analysis was performed to detect the expressions of phospho-JNK, phospho-p38 and phospho-ERK 1/2 in A549, A549/Gem and A549/Gem-CXCR4 cells.Results Gemcitabine-resistance A549 cell ( A549/Gem) was successful constructed by using continuous stepwise exposure to Gemcitabine in vitro.The expression level of CXCR4 was up-regulated in A549/Gem cell than in A549 cell.The CXCR4 shRNA vector could significantly decrease CXCR4 expression in A549/Gem cell.The IC50 values of Gemcitabine in A549, A549/Gem and A549/Gem-CXCR4 cell were (0.08 ±0.01)μmol/L, (14.01 ±0.21)μmol/L and (1.84 ±0.61)μmol/L, respectively.The RI value of Gemcitabine was (127.12 ±12.28) in A549/Gem cells, while the value of RI was (27.3 ±0.98) in A549/Gem-CXCR4 cells.The expression level of phospho-JNK, phospho-p38 and phospho-ERK 1/2 were also markedly inhibited in A549/Gem-CXCR4 cell than in A549/Gem cell.Conclusion CXCR4 is up-regulated in A549/Gem cell.Targeting silence CXCR4 can successfully reverse drug-resistance of Gemcitabine in A549/Gem cells, which hints CXCR4 is associated with lung cancer radiation therapy as an effective molecular target.