AIM:To construct Hsp90 antisense RNA eukaryotic expression vector, transfect it into SGC7901 and SGC7901/VCR of MDR-type human gastric cancer cell lines, HCC7402 of human hepatic cancer and Ec109 of human esophageal c...AIM:To construct Hsp90 antisense RNA eukaryotic expression vector, transfect it into SGC7901 and SGC7901/VCR of MDR-type human gastric cancer cell lines, HCC7402 of human hepatic cancer and Ec109 of human esophageal cancer cell lines, and to study the cell cycle distribution of the gene transected cells and their response to chemotherapeutic drugs.METHODS:A 1.03kb cDNA sequence of Hsp90beta was obtained from the primary plasmid phHSP90 by EcoR I and BamH I nuclease digestion and was cloned to the EcoR I and BamH I site of the pcDNA by T4DNA ligase and an antisense orientation of Hsp90beta expression vector was constructed. The constructs were transfected with lipofectamine and positive clones were selected with G418. The expression of RNA was determined with dot blotting and RNase protection assay, and the expression of Hsp90 protein determined with western blot. Cell cycle distribution of the transfectants was analyzed with flow cytometry, and the drug sensitivity of the transfectants to Adriamycin (ADR), vincrinstine (VCR), mitomycin (MMC) and cyclophosphamide (CTX) with MTT and intracellular drug concentration of the transfectants was determined with flow cytometry.RESULTS:In EcoR I and BamH I restriction analysis, the size and the direction of the cloned sequence of Hsp90beta remained what had been designed and the gene constructs were named pcDNA-Hsp90.AH-SGC7901, AH-SGC7901/VCR, AH-HCC7402 and AH-Ec109 cell clones all expressed Hsp90 anti-sense RNA. The expression of Hsp90 was down-regulated in AH-SGC7901, AH-SGC7901/VCR, AH-HCC7402 and AH-Ec109 cell clones. Cell cycle distribution was changed differently. In AH-SGC7901/VCR and AH-Ec109 cells, G(1) phase cells were increased; S phase and G(2) phase cells were decreased as compared with their parental cell lines. In AH-SGC7901 cell, G(1)phase cells were decreased, G(2) phase cells increased and S phase cells were not changed, and in AH-HCC7402 cells G(1), S and G(2) phase cells remained unchanged as compared with their parental cell lines. The sensitivity of展开更多
AIM:To investigate the anti-tumor effects of Paris chinensis dioscin(PCD)and mechanisms regarding cell cycle regulation and apoptosis in human gastric cancer SGC-7901 cells.METHODS:Cell viability was analyzed by the 3...AIM:To investigate the anti-tumor effects of Paris chinensis dioscin(PCD)and mechanisms regarding cell cycle regulation and apoptosis in human gastric cancer SGC-7901 cells.METHODS:Cell viability was analyzed by the 3-(4,5-di-methylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay.Cell apoptosis was evaluated by flow cytometry and laser scanning confocal microscope(LSCM)using Annexin-V/propidium iodide(PI)staining,and the cell cycle was evaluated using PI staining with flow cytom-etry.Intracellular calcium ions were detected under fluorescence microscope.The expression of cell cycle and apoptosis-related proteins cyclin B1,CDK1,cytochrome C and caspase-3 was measured by immunohistochemical staining.RESULTS:PCD had an anti-proliferation effect on human gastric cancer SGC-7901 cells in a dose-and time-de-pendent manner.After treatment of SGC-7901 cells with PCD,apoptosis appeared in SGC-7901 cells.Morpho-logical changes typical of apoptosis were also observed with LSCM by Annexin V/PI staining,and the cell number of the G0/G1 phase was decreased,while the number of cells in the G2/M phase was increased.Cell cycle-related proteins,such as cyclin B1 and CDK1,were all down-regulated,but caspase-3 and cytochrome C were up-regulated.Moreover,intracellular calcium accumulation occurred in PCD-treated cells.CONCLUSION:G2/M phase arrest and apoptosis induced by PCD are associated with the inhibition of CDK-activating kinase activity and the activation of Ca2+-related mitochondrion pathway in SGC-7901 cells.展开更多
AIM:To gain new insights into tumor metabolism and to identify possible biomarkers with potential diagnostic values to predict tumor metastasis.METHODS:Human gastric cancer SGC-7901 cells were implanted into 24 severe...AIM:To gain new insights into tumor metabolism and to identify possible biomarkers with potential diagnostic values to predict tumor metastasis.METHODS:Human gastric cancer SGC-7901 cells were implanted into 24 severe combined immune deficiency (SCID) mice,which were randomly divided into metastasis group (n=8),non-metastasis group (n=8),and normal group (n=8).Urinary metabolomic information was obtained by gas chromatography/mass spectrometry (GC/MS).RESULTS:There were significant metabolic differences among the three groups (t test,P < 0.05).Ten selected metabolites were different between normal and cancer groups (non-metastasis and metastasis groups),and seven metabolites were also different between non-metastasis and metastasis groups.Two diagnostic models for gastric cancer and metastasis were constructed respectively by the principal component analysis (PCA).These PCA models were confirmed by corresponding receiver operating characteristic analysis (area under the curve=1.00).CONCLUSION:The urinary metabolomic profile is different,and the selected metabolites might be instructive to clinical diagnosis or screening metastasis for gastric cancer.展开更多
基金Project supported by the National Natural Science Foundation of China,No.39570806National Excel1ent Youth Scientific Foundation,No.3952020.
文摘AIM:To construct Hsp90 antisense RNA eukaryotic expression vector, transfect it into SGC7901 and SGC7901/VCR of MDR-type human gastric cancer cell lines, HCC7402 of human hepatic cancer and Ec109 of human esophageal cancer cell lines, and to study the cell cycle distribution of the gene transected cells and their response to chemotherapeutic drugs.METHODS:A 1.03kb cDNA sequence of Hsp90beta was obtained from the primary plasmid phHSP90 by EcoR I and BamH I nuclease digestion and was cloned to the EcoR I and BamH I site of the pcDNA by T4DNA ligase and an antisense orientation of Hsp90beta expression vector was constructed. The constructs were transfected with lipofectamine and positive clones were selected with G418. The expression of RNA was determined with dot blotting and RNase protection assay, and the expression of Hsp90 protein determined with western blot. Cell cycle distribution of the transfectants was analyzed with flow cytometry, and the drug sensitivity of the transfectants to Adriamycin (ADR), vincrinstine (VCR), mitomycin (MMC) and cyclophosphamide (CTX) with MTT and intracellular drug concentration of the transfectants was determined with flow cytometry.RESULTS:In EcoR I and BamH I restriction analysis, the size and the direction of the cloned sequence of Hsp90beta remained what had been designed and the gene constructs were named pcDNA-Hsp90.AH-SGC7901, AH-SGC7901/VCR, AH-HCC7402 and AH-Ec109 cell clones all expressed Hsp90 anti-sense RNA. The expression of Hsp90 was down-regulated in AH-SGC7901, AH-SGC7901/VCR, AH-HCC7402 and AH-Ec109 cell clones. Cell cycle distribution was changed differently. In AH-SGC7901/VCR and AH-Ec109 cells, G(1) phase cells were increased; S phase and G(2) phase cells were decreased as compared with their parental cell lines. In AH-SGC7901 cell, G(1)phase cells were decreased, G(2) phase cells increased and S phase cells were not changed, and in AH-HCC7402 cells G(1), S and G(2) phase cells remained unchanged as compared with their parental cell lines. The sensitivity of
文摘目的探讨适集思血糖控制(space glucose control,SGC)对ICU应激性高血糖患者血糖管理质量的影响。方法采取前瞻性交叉对照的类试验研究方法观察2018年1月至2019年1月期间收住苏州大学附属第二医院综合ICU应激性高血糖的患者。以常规血糖管理作为对照组,SGC方式作为试验组;入选患者每24 h两组间互换,试验终点为96 h。比较两种管理方式对患者血糖管理质量指标的差异,主要包括患者的平均血糖值、血糖最高值和最低值、平均血糖监测间隔时间、累积胰岛素用量。采用SPSS 23.0进行统计分析,正态分布的计量资料采用配对t检验;不符合正态分布的计量资料,采用非参数秩和检验;以P<0.05为差异有统计学意义。结果共有41例患者进入研究,SGC方式的血糖水平明显低于常规方式[(8.60±1.42)mmol/L vs(10.02±1.49)mmol/L,P<0.01],高血糖发生次数低于常规方式(16.59±8.56 vs 18.73±7.91,P=0.023),血糖值在目标血糖值范围内的频次明显高于常规血糖管理方式(53.07±19.11 vs 29.44±19.60,P<0.01)。但SGC方式的低血糖发生频次、血糖监测频次、累积胰岛素用量等均高于常规方式,分别为:[1(0,5)vs 0(0,2),P<0.01]、(1.36±0.23 vs 1.89±0.28,P<0.01)、[(139.61±77.06)U vs(107.49±64.41)U,P<0.01)]。结论SGC血糖管理方式能更优化控制血糖在目标血糖范围,但易导致轻度低血糖的发生,并且在一定程度上增加了医护人员的工作量。
基金Supported by The grant from the Department of Education of Shandong Province,China,No.J10LF18
文摘AIM:To investigate the anti-tumor effects of Paris chinensis dioscin(PCD)and mechanisms regarding cell cycle regulation and apoptosis in human gastric cancer SGC-7901 cells.METHODS:Cell viability was analyzed by the 3-(4,5-di-methylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay.Cell apoptosis was evaluated by flow cytometry and laser scanning confocal microscope(LSCM)using Annexin-V/propidium iodide(PI)staining,and the cell cycle was evaluated using PI staining with flow cytom-etry.Intracellular calcium ions were detected under fluorescence microscope.The expression of cell cycle and apoptosis-related proteins cyclin B1,CDK1,cytochrome C and caspase-3 was measured by immunohistochemical staining.RESULTS:PCD had an anti-proliferation effect on human gastric cancer SGC-7901 cells in a dose-and time-de-pendent manner.After treatment of SGC-7901 cells with PCD,apoptosis appeared in SGC-7901 cells.Morpho-logical changes typical of apoptosis were also observed with LSCM by Annexin V/PI staining,and the cell number of the G0/G1 phase was decreased,while the number of cells in the G2/M phase was increased.Cell cycle-related proteins,such as cyclin B1 and CDK1,were all down-regulated,but caspase-3 and cytochrome C were up-regulated.Moreover,intracellular calcium accumulation occurred in PCD-treated cells.CONCLUSION:G2/M phase arrest and apoptosis induced by PCD are associated with the inhibition of CDK-activating kinase activity and the activation of Ca2+-related mitochondrion pathway in SGC-7901 cells.
基金Supported by The Key Program of Science and Technology Commission of Shanghai Municipality,Project No.09JC1411600Natural Science Foundation of Shanghai,No.08ZR1411300
文摘AIM:To gain new insights into tumor metabolism and to identify possible biomarkers with potential diagnostic values to predict tumor metastasis.METHODS:Human gastric cancer SGC-7901 cells were implanted into 24 severe combined immune deficiency (SCID) mice,which were randomly divided into metastasis group (n=8),non-metastasis group (n=8),and normal group (n=8).Urinary metabolomic information was obtained by gas chromatography/mass spectrometry (GC/MS).RESULTS:There were significant metabolic differences among the three groups (t test,P < 0.05).Ten selected metabolites were different between normal and cancer groups (non-metastasis and metastasis groups),and seven metabolites were also different between non-metastasis and metastasis groups.Two diagnostic models for gastric cancer and metastasis were constructed respectively by the principal component analysis (PCA).These PCA models were confirmed by corresponding receiver operating characteristic analysis (area under the curve=1.00).CONCLUSION:The urinary metabolomic profile is different,and the selected metabolites might be instructive to clinical diagnosis or screening metastasis for gastric cancer.
