目的:探讨小檗胺对 NB4白血病细胞生长的影响及其机制。方法:体外培养 NB4人白血病细胞,经不同浓度小檗胺处理不同时间后,MIT 法检测药物对 NB4细胞增殖的抑制作用;细胞形态学观察和 DNA 琼脂糖电泳检测细胞凋亡;流式细胞仪检测 DNA 含...目的:探讨小檗胺对 NB4白血病细胞生长的影响及其机制。方法:体外培养 NB4人白血病细胞,经不同浓度小檗胺处理不同时间后,MIT 法检测药物对 NB4细胞增殖的抑制作用;细胞形态学观察和 DNA 琼脂糖电泳检测细胞凋亡;流式细胞仪检测 DNA 含量及周期;巢式 PCR 检测 PML/RARα融合基因表达。结果:NB4细胞经不同浓度小檗胺处理不同时间后细胞生长受抑,并呈时间剂量依赖性,48 h IC_(50)为3.86μg·mL^(-1);细胞形态学观察到细胞凋亡特征,DNA 琼脂糖电泳显示典型的凋亡梯形图,经12μg·mL^(-1)的小檗胺处理48 h 后流式细胞仪检测发现细胞凋亡率明显增加,从2.83%增加58.44%;但未发现药物作用后 NB4细胞 PML/RARα基因表达水平的改变。结论:小檗胺能抑制 NB4细胞生长,其机制之一可能是诱导细胞凋亡。展开更多
目的：研究连花清瘟胶囊抗肿瘤的药理作用。方法应用MTT法检测连花清瘟胶囊对乳腺癌 MCF－7细胞增殖的抑制程度，AO／EB双荧光染色法检测细胞凋亡形态，DNA琼脂糖凝胶电泳观察细胞 DNA 片段。结果在浓度为200～2667μg· mL －1内...目的：研究连花清瘟胶囊抗肿瘤的药理作用。方法应用MTT法检测连花清瘟胶囊对乳腺癌 MCF－7细胞增殖的抑制程度，AO／EB双荧光染色法检测细胞凋亡形态，DNA琼脂糖凝胶电泳观察细胞 DNA 片段。结果在浓度为200～2667μg· mL －1内，连花清瘟胶囊对乳腺癌MCF－7细胞的增殖有显著的抑制作用，其抑制作用随药物浓度的增加而增强，对MCF－7细胞增殖抑制率由10．50％增加到43．83％，与对照组比较呈显著差异性（P＜0．05vs正常组）。连花清瘟胶囊明显影响乳腺癌MCF－7细胞的染色质，发生固缩等凋亡形态的变化及细胞凋亡的 DNA梯带形状。结论连花清瘟胶囊具有抑制乳腺癌细胞增殖和诱导其凋亡的抗肿瘤的药理作用。展开更多
The MaMV-DC cyanophage,which infects the bloom-forming cyanobacterium Microcystis aeruginosa,was isolated from Lake Dianchi,Kunming,China.Twenty-one cyanobacterial strains were used to detect the host range of MaMV-DC...The MaMV-DC cyanophage,which infects the bloom-forming cyanobacterium Microcystis aeruginosa,was isolated from Lake Dianchi,Kunming,China.Twenty-one cyanobacterial strains were used to detect the host range of MaMV-DC.Microcystic aeruginosa FACHB-524 and plaque purification were used to isolate individual cyanophages,and culturing MaMV-DC with cyanobacteria allowed us to prepare purified cyanophages for further analysis.Electron microscopy demonstrated that the negatively stained viral particles are tadpole-shaped with an icosahedral head approximately 70 nm in diameter and a contractile tail approximately 160 nm in length.Using one-step growth experiments,the latent period and burst size of MaMV-DC were estimated to be 24–48 hours and approximately 80infectious units per cell,respectively.Restriction endonuclease digestion and agarose gel electrophoresis were performed using purified MaMV-DC genomic DNA,and the genome size was estimated to be approximately 160 kb.Sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE)analysis revealed four major structural proteins.These results support the growing interest in using freshwater cyanophages to control bloom-forming cyanobacterium.展开更多
AIM： To investigate the effect of 5-allyl-7-gen-difluoromethylenechrysin （ADFMChR） on apoptosis of human liver carcinoma HepG2 cell line and the molecular mechanisms involved.METHODS： HepG2 cells and L-02 cells we...AIM： To investigate the effect of 5-allyl-7-gen-difluoromethylenechrysin （ADFMChR） on apoptosis of human liver carcinoma HepG2 cell line and the molecular mechanisms involved.METHODS： HepG2 cells and L-02 cells were cultured in vitro and the inhibitory effect of ADFMChR on their proliferation was measured by MTT assay. The apoptosis of HepG2 cells was determined by flow cytometry （FCM） using propidium iodide （PI） fluorescence staining. DNA ladder bands were observed by DNA agarose gel electrophoresis. The influence of ADFMChR on the proxisome proliferator-activated receptor γ （PPARγ）, NF-κB, Bcl-2 and Bax protein expression of HepG2 cells were analyzed by Western blotting.RESULTS： MTT assay showed that ADFMChR significantly inhibited proliferation of HepG2 cells in a dose- dependent manner, with little effect on growth of L-02 cells, and when ICs0 was measured as 8.45 μmol/L and 191.55 μmol/L respectively, the potency of ADFMChR to HepG2 cells, was found to be similar to 5-fluorouracil （5-FU, ICso was 9.27 μmol/L）. The selective index of ADFMChR cytotoxicity to HepG2 cells was 22.67 （191.55/8.45）, higher than 5-FU （SI was 7.05 （65.37/9.27）. FCM with PI staining demonstrated that the apoptosis rates of HepG2 cells treated with 3.0, 10.0 and 30.0 μmol/L ADFMChR for 48 h were 5.79%, 9.29% and 37.8%, respectively, and were significantly higher when treated with 30.0 μmol/L ADFMChR than when treated with 30.0 μmol/L ChR （16.0%） （P 〈 0.05） and were similar to those obtained with 30.0 μmol/L 5-FU（41.0%）. DNA agarose gel electrophoresis showed that treatment of HepG2 cells with 10.0 μmol/L ADFMChR for 48 h and 72 h resulted in typical DNA ladders which could be reversed by 10.00 pmol/1 GW9662, a blocker of PPARy. Western blotting analysis revealed that aEer 24 h of treatment with 3.0, 10.0, 30.0 μmol/L ADFMChR, PPARy and Bax protein expression in HepG2 cells increased but Bcl-2 and NF-κB expression decreased; however, pre-incubation with 10.0 μmol/L GW9662 could efficiently antagonize and weaken the regulatory effect of 3.0, 30.0 μmol/L ADFMChR on PPARy and NF-KB protein expression in HepG2 cells.CONCLUSION： ADFMChR induces apoptosis of HepG2 cell lines by activating PPARγ, inhibiting protein expression of Bcl-2 and NF-κB, and increasing Bax expression.展开更多
Objective： To investigate the association of XRCC4 polymorphic variants at G-1394T （rs6869366） with colorectal cancer susceptibility. Methods： In this hospital-based case-control study, the association of XRCC4 po...Objective： To investigate the association of XRCC4 polymorphic variants at G-1394T （rs6869366） with colorectal cancer susceptibility. Methods： In this hospital-based case-control study, the association of XRCC4 polymorphism with colorectal cancer risk in Chinese population was investigated. In total, 171 patients with colorectal cancer and 171 healthy individuals matched for age and gender were selected. The genomic DNAs of the patients and controls were extracted from peripheral blood and the 300 bp target DNA was amplified with Polymerase Chain Reaction. The products were then digested with restriction endonuclease HinclI, followed by agarose electrophoresis to identify the genotype. Results： We found a significant difference in the frequency of the XRCC4 G-1394T genotype between the colorectal cancer and control groups in female （1/127 vs 8/122, P〈0.05）. Those with G/T at XRCC4 G-1394T showed a decreased risk of colorectal cancer susceptibility compared with those with T/T （OR 0.113, 95%CI 0.014-0.932）. However, in overall population or in male, there was no significant difference of the distribution between the colorectal cancer and control groups. Conclusion： Our findings with decreased risk of colorectal cancer susceptibility suggested that the G allele of XRCC4 G-1394T were associated in female.展开更多
文摘目的:探讨小檗胺对 NB4白血病细胞生长的影响及其机制。方法:体外培养 NB4人白血病细胞,经不同浓度小檗胺处理不同时间后,MIT 法检测药物对 NB4细胞增殖的抑制作用;细胞形态学观察和 DNA 琼脂糖电泳检测细胞凋亡;流式细胞仪检测 DNA 含量及周期;巢式 PCR 检测 PML/RARα融合基因表达。结果:NB4细胞经不同浓度小檗胺处理不同时间后细胞生长受抑,并呈时间剂量依赖性,48 h IC_(50)为3.86μg·mL^(-1);细胞形态学观察到细胞凋亡特征,DNA 琼脂糖电泳显示典型的凋亡梯形图,经12μg·mL^(-1)的小檗胺处理48 h 后流式细胞仪检测发现细胞凋亡率明显增加,从2.83%增加58.44%;但未发现药物作用后 NB4细胞 PML/RARα基因表达水平的改变。结论:小檗胺能抑制 NB4细胞生长,其机制之一可能是诱导细胞凋亡。
文摘目的：研究连花清瘟胶囊抗肿瘤的药理作用。方法应用MTT法检测连花清瘟胶囊对乳腺癌 MCF－7细胞增殖的抑制程度，AO／EB双荧光染色法检测细胞凋亡形态，DNA琼脂糖凝胶电泳观察细胞 DNA 片段。结果在浓度为200～2667μg· mL －1内，连花清瘟胶囊对乳腺癌MCF－7细胞的增殖有显著的抑制作用，其抑制作用随药物浓度的增加而增强，对MCF－7细胞增殖抑制率由10．50％增加到43．83％，与对照组比较呈显著差异性（P＜0．05vs正常组）。连花清瘟胶囊明显影响乳腺癌MCF－7细胞的染色质，发生固缩等凋亡形态的变化及细胞凋亡的 DNA梯带形状。结论连花清瘟胶囊具有抑制乳腺癌细胞增殖和诱导其凋亡的抗肿瘤的药理作用。
基金National Natural Science Foundation of China(grant nos.31072239,31270213)Knowledge Innovation Program of the Chinese Academy of Sciences(grant no.KSCX2-EW-Z-3)StateKey Laboratory of Freshwater Ecology&Biotechnology Program(grant no.2011FBZ12)
文摘The MaMV-DC cyanophage,which infects the bloom-forming cyanobacterium Microcystis aeruginosa,was isolated from Lake Dianchi,Kunming,China.Twenty-one cyanobacterial strains were used to detect the host range of MaMV-DC.Microcystic aeruginosa FACHB-524 and plaque purification were used to isolate individual cyanophages,and culturing MaMV-DC with cyanobacteria allowed us to prepare purified cyanophages for further analysis.Electron microscopy demonstrated that the negatively stained viral particles are tadpole-shaped with an icosahedral head approximately 70 nm in diameter and a contractile tail approximately 160 nm in length.Using one-step growth experiments,the latent period and burst size of MaMV-DC were estimated to be 24–48 hours and approximately 80infectious units per cell,respectively.Restriction endonuclease digestion and agarose gel electrophoresis were performed using purified MaMV-DC genomic DNA,and the genome size was estimated to be approximately 160 kb.Sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE)analysis revealed four major structural proteins.These results support the growing interest in using freshwater cyanophages to control bloom-forming cyanobacterium.
基金Supported by Research Grant of Department of Science and Technology of Hunan Province,2007TP4017
文摘AIM： To investigate the effect of 5-allyl-7-gen-difluoromethylenechrysin （ADFMChR） on apoptosis of human liver carcinoma HepG2 cell line and the molecular mechanisms involved.METHODS： HepG2 cells and L-02 cells were cultured in vitro and the inhibitory effect of ADFMChR on their proliferation was measured by MTT assay. The apoptosis of HepG2 cells was determined by flow cytometry （FCM） using propidium iodide （PI） fluorescence staining. DNA ladder bands were observed by DNA agarose gel electrophoresis. The influence of ADFMChR on the proxisome proliferator-activated receptor γ （PPARγ）, NF-κB, Bcl-2 and Bax protein expression of HepG2 cells were analyzed by Western blotting.RESULTS： MTT assay showed that ADFMChR significantly inhibited proliferation of HepG2 cells in a dose- dependent manner, with little effect on growth of L-02 cells, and when ICs0 was measured as 8.45 μmol/L and 191.55 μmol/L respectively, the potency of ADFMChR to HepG2 cells, was found to be similar to 5-fluorouracil （5-FU, ICso was 9.27 μmol/L）. The selective index of ADFMChR cytotoxicity to HepG2 cells was 22.67 （191.55/8.45）, higher than 5-FU （SI was 7.05 （65.37/9.27）. FCM with PI staining demonstrated that the apoptosis rates of HepG2 cells treated with 3.0, 10.0 and 30.0 μmol/L ADFMChR for 48 h were 5.79%, 9.29% and 37.8%, respectively, and were significantly higher when treated with 30.0 μmol/L ADFMChR than when treated with 30.0 μmol/L ChR （16.0%） （P 〈 0.05） and were similar to those obtained with 30.0 μmol/L 5-FU（41.0%）. DNA agarose gel electrophoresis showed that treatment of HepG2 cells with 10.0 μmol/L ADFMChR for 48 h and 72 h resulted in typical DNA ladders which could be reversed by 10.00 pmol/1 GW9662, a blocker of PPARy. Western blotting analysis revealed that aEer 24 h of treatment with 3.0, 10.0, 30.0 μmol/L ADFMChR, PPARy and Bax protein expression in HepG2 cells increased but Bcl-2 and NF-κB expression decreased; however, pre-incubation with 10.0 μmol/L GW9662 could efficiently antagonize and weaken the regulatory effect of 3.0, 30.0 μmol/L ADFMChR on PPARy and NF-KB protein expression in HepG2 cells.CONCLUSION： ADFMChR induces apoptosis of HepG2 cell lines by activating PPARγ, inhibiting protein expression of Bcl-2 and NF-κB, and increasing Bax expression.
文摘Objective： To investigate the association of XRCC4 polymorphic variants at G-1394T （rs6869366） with colorectal cancer susceptibility. Methods： In this hospital-based case-control study, the association of XRCC4 polymorphism with colorectal cancer risk in Chinese population was investigated. In total, 171 patients with colorectal cancer and 171 healthy individuals matched for age and gender were selected. The genomic DNAs of the patients and controls were extracted from peripheral blood and the 300 bp target DNA was amplified with Polymerase Chain Reaction. The products were then digested with restriction endonuclease HinclI, followed by agarose electrophoresis to identify the genotype. Results： We found a significant difference in the frequency of the XRCC4 G-1394T genotype between the colorectal cancer and control groups in female （1/127 vs 8/122, P〈0.05）. Those with G/T at XRCC4 G-1394T showed a decreased risk of colorectal cancer susceptibility compared with those with T/T （OR 0.113, 95%CI 0.014-0.932）. However, in overall population or in male, there was no significant difference of the distribution between the colorectal cancer and control groups. Conclusion： Our findings with decreased risk of colorectal cancer susceptibility suggested that the G allele of XRCC4 G-1394T were associated in female.
