AIM:To investigate whether the apoptotic activities of 8-bromo-7-methoxychrysin(BrMC) involve reactive oxygen species(ROS) generation and c-Jun N-terminal kinase(JNK) activation in human hepatocellular carcinoma cells...AIM:To investigate whether the apoptotic activities of 8-bromo-7-methoxychrysin(BrMC) involve reactive oxygen species(ROS) generation and c-Jun N-terminal kinase(JNK) activation in human hepatocellular carcinoma cells(HCC).METHODS:HepG2,Bel-7402 and L-02 cell lines were cultured in vitro and the apoptotic effects of BrMC were evaluated by flow cytometry(FCM) after propidium iodide(PI) staining,caspase-3 activity using enzymelinked immunosorbent assay(ELISA),and DNA agarose gel electrophoresis.ROS production was evaluated by FCM after dichlorodihydrofluorescein diacetate(DCHFDA) probe labeling.The phosphorylation level of JNK and c-Jun protein was analyzed by Western blotting.RESULTS:FCM after PI staining showed a dose-dependent increase in the percentage of the sub-G1 cell pop-ulation(P < 0.05),reaching 39.0% ± 2.8% of HepG2 cells after 48 h of treatment with BrMC at 10 μmol/L.The potency of BrMC to HepG2 and Bel-7402(32.1% ± 2.6%) cells was found to be more effective than the lead compound,chrysin(16.2% ± 1.6% for HepG2 cells and 11.0% ± 1.3% for Bel-7402 cell) at 40 μmol/L and similar to 5-flurouracil(33.0% ± 2.1% for HepG2 cells and 29.3% ± 2.3% for Bel-7402 cells) at 10 μmol/L.BrMC had little effect on human embryo liver L-02 cells,with the percentage of sub-G1 cell population 5.4% ± 1.8%.Treatment of HepG2 cells with BrMC for 48 h also increased the levels of active caspase-3,in a concentration-dependent manner.z-DEVD-fmk,a caspase-3specific inhibitor,prevented the activation of caspase-3.Treatment with BrMC at 10 μmol/L for 48 h resulted in the formation of a DNA ladder.Treatment of cells with BrMC(10 μmol/L) increased mean fluorescence intensity of DCHF-DA in HepG2 cells from 7.2 ± 1.12 at 0 h to 79.8 ± 3.9 at 3 h and 89.7 ± 4.7 at 6 h.BrMC did not affect ROS generation in L-02 cells.BrMC treatment failed to induce cell death and caspase-3 activation in HepG2 cells pretreated with N-acetylcysteine(10 mmol/L).In addition,in HepG2 cells treated with BrMC(2.5,5.0,10.0 μmol/L) for 12 h,JNK acti展开更多
Background The global strategy for the diagnosis, management and prevention of chronic obstructive pulmonary disease （COPD） guidelines classify patients into four groups according to the number of symptoms and the l...Background The global strategy for the diagnosis, management and prevention of chronic obstructive pulmonary disease （COPD） guidelines classify patients into four groups according to the number of symptoms and the level of future risk of acute exacerbation COPD （AECOPD）. This study aimed to compare the results of different methods used in diagnosis of COPD and evaluate the accuracy of the assessment methods in guiding clinical practice. Methods A survey was conducted of 194 COPD outpatients between March and September 2012. Demographic characteristics, the number of exacerbations the patient has had within the previous 12 months, COPD assessment test （CAT）, Modified British Medical Research Council （mMRC） scale, and results of the lung function tests were recorded. Results Of the 194 patients assessed, 21 had a CAT score 〉10 and an mMRC grade ≤1, 13 had a CAT score 〈10 and an mMRC grade ≥2. A predicted forced expiratory volume in one second （FEV1%） of 〈50% with less than two acute exacerbations was observed in 39 patients, while a predicted FEV1% of 〉50% was noted in 20 patients with two or more acute exacerbations. The sensitivity of a predicted FEV1% 〈50% in predicting the risk of AECOPD in the future was 80.9%, while that in the real number of AECOPD events recorded was 62.8%, the difference being statistically significant （P=0.004）. The sensitivity of CAT in predicting the severity of symptoms was 90%, while that of mMRC was 83.8%, and the difference was not statistically significant. Conclusions The COPD assessment method recommended by the global initiative for chronic obstructive pulmonary disease （GOLD） 2011 is complicated and should be simplified. CAT is more comprehensive and accurate than mMRC. The lung function classification is a better tool for predicting the risk of AECOPD in the future, and the number of AECOPD can be referred to when required.展开更多
基金Supported by Hunan Provincial Natural Science Foundation of China,No.03JJY5009
文摘AIM:To investigate whether the apoptotic activities of 8-bromo-7-methoxychrysin(BrMC) involve reactive oxygen species(ROS) generation and c-Jun N-terminal kinase(JNK) activation in human hepatocellular carcinoma cells(HCC).METHODS:HepG2,Bel-7402 and L-02 cell lines were cultured in vitro and the apoptotic effects of BrMC were evaluated by flow cytometry(FCM) after propidium iodide(PI) staining,caspase-3 activity using enzymelinked immunosorbent assay(ELISA),and DNA agarose gel electrophoresis.ROS production was evaluated by FCM after dichlorodihydrofluorescein diacetate(DCHFDA) probe labeling.The phosphorylation level of JNK and c-Jun protein was analyzed by Western blotting.RESULTS:FCM after PI staining showed a dose-dependent increase in the percentage of the sub-G1 cell pop-ulation(P < 0.05),reaching 39.0% ± 2.8% of HepG2 cells after 48 h of treatment with BrMC at 10 μmol/L.The potency of BrMC to HepG2 and Bel-7402(32.1% ± 2.6%) cells was found to be more effective than the lead compound,chrysin(16.2% ± 1.6% for HepG2 cells and 11.0% ± 1.3% for Bel-7402 cell) at 40 μmol/L and similar to 5-flurouracil(33.0% ± 2.1% for HepG2 cells and 29.3% ± 2.3% for Bel-7402 cells) at 10 μmol/L.BrMC had little effect on human embryo liver L-02 cells,with the percentage of sub-G1 cell population 5.4% ± 1.8%.Treatment of HepG2 cells with BrMC for 48 h also increased the levels of active caspase-3,in a concentration-dependent manner.z-DEVD-fmk,a caspase-3specific inhibitor,prevented the activation of caspase-3.Treatment with BrMC at 10 μmol/L for 48 h resulted in the formation of a DNA ladder.Treatment of cells with BrMC(10 μmol/L) increased mean fluorescence intensity of DCHF-DA in HepG2 cells from 7.2 ± 1.12 at 0 h to 79.8 ± 3.9 at 3 h and 89.7 ± 4.7 at 6 h.BrMC did not affect ROS generation in L-02 cells.BrMC treatment failed to induce cell death and caspase-3 activation in HepG2 cells pretreated with N-acetylcysteine(10 mmol/L).In addition,in HepG2 cells treated with BrMC(2.5,5.0,10.0 μmol/L) for 12 h,JNK acti
文摘Background The global strategy for the diagnosis, management and prevention of chronic obstructive pulmonary disease （COPD） guidelines classify patients into four groups according to the number of symptoms and the level of future risk of acute exacerbation COPD （AECOPD）. This study aimed to compare the results of different methods used in diagnosis of COPD and evaluate the accuracy of the assessment methods in guiding clinical practice. Methods A survey was conducted of 194 COPD outpatients between March and September 2012. Demographic characteristics, the number of exacerbations the patient has had within the previous 12 months, COPD assessment test （CAT）, Modified British Medical Research Council （mMRC） scale, and results of the lung function tests were recorded. Results Of the 194 patients assessed, 21 had a CAT score 〉10 and an mMRC grade ≤1, 13 had a CAT score 〈10 and an mMRC grade ≥2. A predicted forced expiratory volume in one second （FEV1%） of 〈50% with less than two acute exacerbations was observed in 39 patients, while a predicted FEV1% of 〉50% was noted in 20 patients with two or more acute exacerbations. The sensitivity of a predicted FEV1% 〈50% in predicting the risk of AECOPD in the future was 80.9%, while that in the real number of AECOPD events recorded was 62.8%, the difference being statistically significant （P=0.004）. The sensitivity of CAT in predicting the severity of symptoms was 90%, while that of mMRC was 83.8%, and the difference was not statistically significant. Conclusions The COPD assessment method recommended by the global initiative for chronic obstructive pulmonary disease （GOLD） 2011 is complicated and should be simplified. CAT is more comprehensive and accurate than mMRC. The lung function classification is a better tool for predicting the risk of AECOPD in the future, and the number of AECOPD can be referred to when required.
