AIM: To investigate the effects of Recql5 deficiency on liver injury induced by lipopolysaccharide/D-galactosamine(LPS/D-Gal).METHODS: Liver injury was induced in wild type(WT) or Recql5-deficient mice using LPS/D-Gal...AIM: To investigate the effects of Recql5 deficiency on liver injury induced by lipopolysaccharide/D-galactosamine(LPS/D-Gal).METHODS: Liver injury was induced in wild type(WT) or Recql5-deficient mice using LPS/D-Gal,and assessed by histological,serum transaminases,and mortality analyses. Hepatocellular apoptosis was quantified by transferase d UTP nick end labeling assay and Westernblot analysis of cleaved caspase-3. Liver inflammatory chemokine and cytochrome P450 expression was analyzed by quantitative reverse transcription-PCR. Neutrophil infiltration was evaluated by myeloperoxidase activity. Expression and phosphorylation of ERK,JNK,p65,and H2 A.X was determined by Western blot. Oxidative stress was evaluated by measuring malondialdehyde production and nitric oxide synthase,superoxide dismutase,glutathione peroxidase,catalase,and glutathione reductase activity.RESULTS: following LPS/D-Gal exposure,Recql5-deficient mice exhibited enhanced liver injury,as evidenced by more severe hepatic hemorrhage,higher serum aspartate transaminase and alanine transaminase levels,and lower survival rate. As compared to WT mice,Recql5-deficient mice showed an increased number of apoptotic hepatocytes and higher cleaved caspase-3 levels. Recql5-deficient mice exhibited increased DNA damage,as evidenced by increased γ-H2 A.X levels. Inflammatory cytokine levels,neutrophil infiltration,and ERK phosphorylation were also significantly increased in the knockout mice. Additionally,Recql5-deficicent mice exhibited increased malondialdehyde production and elevated inducible nitric oxide synthase,superoxide dismutase,glutathione peroxidase,catalase,and glutathione reductase activity,indicative of enhanced oxidative stress. Moreover,CYP450 expression was significantly downregulated in Recql5-deficient mice after LPS/D-Gal treatment.CONCLUSION: Recql5 protects the liver against LPS/D-Gal-induced injury through suppression of hepatocyte apoptosis and oxidative stress and modulation of CYP450 expression.展开更多
AIM:To investigate the effects of phosphatase and tensin homolog deleted on chromosome 10(PTEN) deficiency on the cytotoxicity of chemotherapeutic agents toward colorectal cancer cells.METHODS:PTEN-deficient colorecta...AIM:To investigate the effects of phosphatase and tensin homolog deleted on chromosome 10(PTEN) deficiency on the cytotoxicity of chemotherapeutic agents toward colorectal cancer cells.METHODS:PTEN-deficient colorectal cancer(CRC) cells were generated by human somatic cell gene targeting using the adeno-associated virus system. The cytotoxic effects of compounds including curcumin,5-fluorouracil(5-FU),dihydroartemisinin(DHA),irinotecan(CPT-11)and oxaliplatin(OXA) on cancer cells were determined using the MTT assay. Enhanced cytotoxicity of curcumin in PTEN-deficient CRC cells was observed,and this was confirmed using clonogenic assays. Apoptosis and cell cycle progression were analyzed by flow cytometry.Levels of apoptosis and cell cycle-related proteins were examined by Western blotting.RESULTS:We developed an isogenic set of CRC cell lines that differed only in their PTEN status. Using this set of cell lines,we found that disruption of the PTEN gene had no effect on the sensitivity of CRC cells to5-FU,CPT-11,DHA,or OXA,whereas PTEN disruption increased the sensitivity of CRC cells to curcumin. Loss of PTEN did not alter the curcumin-induced apoptosis in CRC cells. However,PTEN deficiency led to an altered pattern of curcumin-mediated cell cycle arrest.In HCT116 PTEN+/+cells,curcumin caused a G2/M phase arrest,whereas it caused a G0/G1 phase arrest in HCT116 PTEN-/-cells. Levels of cell cycle-related proteins were consistent with these respective patterns of cell cycle arrest.CONCLUSION:Curcumin shows enhanced cytotoxicity toward PTEN-deficient cancer cells,suggesting that it might be a potential chemotherapeutic agent for cancers harboring PTEN mutations.展开更多
Currently,tuberculosis(TB)is the second most lethal disease in the world caused by a single infectious pathogen.Rapid diagnosis of TB is of great importance for its treatment and management.Xpert MTB/RIF is a novel ra...Currently,tuberculosis(TB)is the second most lethal disease in the world caused by a single infectious pathogen.Rapid diagnosis of TB is of great importance for its treatment and management.Xpert MTB/RIF is a novel rapid diagnostic assay for the diagnosis of pulmonary TB(PTB).Use of the Xpert assay based on bronchoalveolar lavage fluid(BALF)samples is indicated when TB is suspected and sputum smears or cultures are negative.The aim of this meta-analysis was to systematically evaluate the diagnostic performance of the Xpert assay based on BALF samples for the diagnosis of PTB.A systematic review of previously published articles was performed,and rel-evant data were extracted.Meta-DiSc 1.4 and Stata 12.0 were used to analyze the data.When Mycobacterium tuberculosis cultures were used as the criterion standard,the combined sensitivity of BALF-based Xpert was 0.89(95%CI,0.87–0.91),the specificity was 0.87(95%CI,0.85-0.88),the positive likelihood ratio was 8.28(95%CI,5.39–12.71),the negative likelihood ratio was 0.14(95%CI,0.10–0.19)and the diagnostic ratio was 84.08(95%CI,42.00–168.31).When composite reference standard was used as the criterion standard,the above observations were 0.69(95%CI,0.67–0.72),0.98(95%CI,0.97–0.98),41.40(95%CI,14.56–117.71),0.28(95%CI,0.21–0.37)and 190.47(95%CI,50.56–717.54),respectively.The area under the summary receiver operating characteristic curve was close to 1 for both.Overall,the Xpert MTB/RIF assay based on BALF samples showed high sensitivity and specificity for the diag-nosis of PTB and seems to be a reliable rapid detection method.展开更多
基金Supported by National Natural Science Foundation of China,No.81101472 and No.81472556(to Liao W),and No.81372490(to Lu X)Zhejiang Provincial Natural Science Foundation,No.LZ14H160003(to Lu X)+2 种基金Zhejiang Provincial Program for the Cultivation of High-Level Innovative Health Talents(to Lu X)National Basic Research Program of China(973 Project),No.2011CB504603Wenzhou Municipal Science and Technology Bureau Foundation,No.Y20110090(to Li H)
文摘AIM: To investigate the effects of Recql5 deficiency on liver injury induced by lipopolysaccharide/D-galactosamine(LPS/D-Gal).METHODS: Liver injury was induced in wild type(WT) or Recql5-deficient mice using LPS/D-Gal,and assessed by histological,serum transaminases,and mortality analyses. Hepatocellular apoptosis was quantified by transferase d UTP nick end labeling assay and Westernblot analysis of cleaved caspase-3. Liver inflammatory chemokine and cytochrome P450 expression was analyzed by quantitative reverse transcription-PCR. Neutrophil infiltration was evaluated by myeloperoxidase activity. Expression and phosphorylation of ERK,JNK,p65,and H2 A.X was determined by Western blot. Oxidative stress was evaluated by measuring malondialdehyde production and nitric oxide synthase,superoxide dismutase,glutathione peroxidase,catalase,and glutathione reductase activity.RESULTS: following LPS/D-Gal exposure,Recql5-deficient mice exhibited enhanced liver injury,as evidenced by more severe hepatic hemorrhage,higher serum aspartate transaminase and alanine transaminase levels,and lower survival rate. As compared to WT mice,Recql5-deficient mice showed an increased number of apoptotic hepatocytes and higher cleaved caspase-3 levels. Recql5-deficient mice exhibited increased DNA damage,as evidenced by increased γ-H2 A.X levels. Inflammatory cytokine levels,neutrophil infiltration,and ERK phosphorylation were also significantly increased in the knockout mice. Additionally,Recql5-deficicent mice exhibited increased malondialdehyde production and elevated inducible nitric oxide synthase,superoxide dismutase,glutathione peroxidase,catalase,and glutathione reductase activity,indicative of enhanced oxidative stress. Moreover,CYP450 expression was significantly downregulated in Recql5-deficient mice after LPS/D-Gal treatment.CONCLUSION: Recql5 protects the liver against LPS/D-Gal-induced injury through suppression of hepatocyte apoptosis and oxidative stress and modulation of CYP450 expression.
基金Supported by The National Natural Science Foundation of China,No.81101472 to Liao WQNo.31071086 to Lu XC
文摘AIM:To investigate the effects of phosphatase and tensin homolog deleted on chromosome 10(PTEN) deficiency on the cytotoxicity of chemotherapeutic agents toward colorectal cancer cells.METHODS:PTEN-deficient colorectal cancer(CRC) cells were generated by human somatic cell gene targeting using the adeno-associated virus system. The cytotoxic effects of compounds including curcumin,5-fluorouracil(5-FU),dihydroartemisinin(DHA),irinotecan(CPT-11)and oxaliplatin(OXA) on cancer cells were determined using the MTT assay. Enhanced cytotoxicity of curcumin in PTEN-deficient CRC cells was observed,and this was confirmed using clonogenic assays. Apoptosis and cell cycle progression were analyzed by flow cytometry.Levels of apoptosis and cell cycle-related proteins were examined by Western blotting.RESULTS:We developed an isogenic set of CRC cell lines that differed only in their PTEN status. Using this set of cell lines,we found that disruption of the PTEN gene had no effect on the sensitivity of CRC cells to5-FU,CPT-11,DHA,or OXA,whereas PTEN disruption increased the sensitivity of CRC cells to curcumin. Loss of PTEN did not alter the curcumin-induced apoptosis in CRC cells. However,PTEN deficiency led to an altered pattern of curcumin-mediated cell cycle arrest.In HCT116 PTEN+/+cells,curcumin caused a G2/M phase arrest,whereas it caused a G0/G1 phase arrest in HCT116 PTEN-/-cells. Levels of cell cycle-related proteins were consistent with these respective patterns of cell cycle arrest.CONCLUSION:Curcumin shows enhanced cytotoxicity toward PTEN-deficient cancer cells,suggesting that it might be a potential chemotherapeutic agent for cancers harboring PTEN mutations.
文摘Currently,tuberculosis(TB)is the second most lethal disease in the world caused by a single infectious pathogen.Rapid diagnosis of TB is of great importance for its treatment and management.Xpert MTB/RIF is a novel rapid diagnostic assay for the diagnosis of pulmonary TB(PTB).Use of the Xpert assay based on bronchoalveolar lavage fluid(BALF)samples is indicated when TB is suspected and sputum smears or cultures are negative.The aim of this meta-analysis was to systematically evaluate the diagnostic performance of the Xpert assay based on BALF samples for the diagnosis of PTB.A systematic review of previously published articles was performed,and rel-evant data were extracted.Meta-DiSc 1.4 and Stata 12.0 were used to analyze the data.When Mycobacterium tuberculosis cultures were used as the criterion standard,the combined sensitivity of BALF-based Xpert was 0.89(95%CI,0.87–0.91),the specificity was 0.87(95%CI,0.85-0.88),the positive likelihood ratio was 8.28(95%CI,5.39–12.71),the negative likelihood ratio was 0.14(95%CI,0.10–0.19)and the diagnostic ratio was 84.08(95%CI,42.00–168.31).When composite reference standard was used as the criterion standard,the above observations were 0.69(95%CI,0.67–0.72),0.98(95%CI,0.97–0.98),41.40(95%CI,14.56–117.71),0.28(95%CI,0.21–0.37)and 190.47(95%CI,50.56–717.54),respectively.The area under the summary receiver operating characteristic curve was close to 1 for both.Overall,the Xpert MTB/RIF assay based on BALF samples showed high sensitivity and specificity for the diag-nosis of PTB and seems to be a reliable rapid detection method.
