To investigate the genotoxicity and reveal the potential toxicological mechanisms of Hexabromocyclododecane （HBCD）, human breast cells HBL-100 were exposed to a sequence of HBCD concentrations （0, 5, 10, and 50 mg/...To investigate the genotoxicity and reveal the potential toxicological mechanisms of Hexabromocyclododecane （HBCD）, human breast cells HBL-100 were exposed to a sequence of HBCD concentrations （0, 5, 10, and 50 mg/L） for 24 h. With a series of zymology and molecular biology methods, we found that HBCD induced dose-dependent oxidative stress on HBL-100 DNA. As revealed in q RT-PCR, activated prognostic factor ATM down-regulated tumor suppressor gene BRCA1 and prompted DNA repair genes h OGG1 and h MTH1 expression in lower concentrations of HBCD （〈 10 mg/L）. However, DNA repair were inhibited as well as cell proliferation rate by higher concentrations of HBCD （50 mg/L）. The results inferred that the genotoxicity of HBCD was dose-dependent and related to DNA repair pathway.展开更多
[ Objective ] To develop a rapid efficient method for detecting mycoplasma contamination in cell cultures. [ Method] A pair of primers was designed according to two highly conserved nucleotide sequences of the 16S RNA...[ Objective ] To develop a rapid efficient method for detecting mycoplasma contamination in cell cultures. [ Method] A pair of primers was designed according to two highly conserved nucleotide sequences of the 16S RNA from six kinds of mycoplasma that commonly contaminated cells. Then the mycoplasma contamination of 25 cell samples was defected by PCR and DNA fluorescence staining. EResultl When these cell samples were detected by DNA fluorescence staining, the positive rate and probable positive rate were respectively 24% and 16%. And when they were detected by PCR, the positive rate was 36%. [ Condusion] The PCR method is more sensitive and specific than the DNA fluorescence staining, and combining these two methods is the optimal way to detect mycoplasma contamination in cell cultures.展开更多
The ＇double T-DNA＇ binary vector p13HSR which harbored two independent T-DNAs, containing hygromycin phosphotransferase gene （hpf） in one T-DNA region and three target genes （hLF, SB401, RZ10） in another T-DNA r...The ＇double T-DNA＇ binary vector p13HSR which harbored two independent T-DNAs, containing hygromycin phosphotransferase gene （hpf） in one T-DNA region and three target genes （hLF, SB401, RZ10） in another T-DNA region, was used to generate selectable marker-free transgenic rice by Agrobacterium-mediated transformation. The regenerated plants with both the three target genes and the selectable marker gene hpt were selected for anther culture. RT-PCR analysis indicated that target genes were inserted in rice genomic DNA and successfully transcribed. It took only one year to obtain double haploid selectable marker-free transgenic plants containing the three target genes with co-transformation followed by anther culture technique, and the efficiency was 12.2%. It was also noted that one or two target genes derived from the binary vector were lost in some transgenic rice plants.展开更多
目的:Siloranes是一种含硅的新型牙科复合树脂环氧乙烷单体,较传统的含碳的环氧乙烷单体具有更好的生物相容性。本研究的目的为用哺乳动物细胞培养检测姐妹染色体交换的方法评估Siloranes的基因毒性,并与传统的环氧乙烷比较。方法:研究...目的:Siloranes是一种含硅的新型牙科复合树脂环氧乙烷单体,较传统的含碳的环氧乙烷单体具有更好的生物相容性。本研究的目的为用哺乳动物细胞培养检测姐妹染色体交换的方法评估Siloranes的基因毒性,并与传统的环氧乙烷比较。方法:研究所用的传统环氧乙烷为CyracureTM UVR6105。Silorane包括Ph-Sil,TET-Sil和二者50/50(w/w)的混合物(3M-ESPE)。用CHO细胞在有或无S9情况下检测姐妹染色体交换(SCE)。Bonferron post hoc统计分析检测有无显著性差异。结果:与溶剂对照相比,在有或无S9情况下,环氧乙烷UVR6105显著性地增加了培养CHO细胞的SCE数目。3种Silorane单体Ph-Sil,TET-Sil或Sil-Mix均未能增加SCE数目。结论:用哺乳动物细胞培养检测姐妹染色体交换的方法评估显示,3种新型Silorane单体Ph-Sil,TET-Sil或Sil-Mix均未检测出基因毒性效果,提示Silorane单体具有较低基因毒性,是适于研发牙科树脂的新型单体。展开更多
基金supported by the National Natural Science Foundation of China(No.41406088)The open fund of Key Laboratory for Ecological Environment in Coastal Areas,State Oceanic Administration(201506)
文摘To investigate the genotoxicity and reveal the potential toxicological mechanisms of Hexabromocyclododecane （HBCD）, human breast cells HBL-100 were exposed to a sequence of HBCD concentrations （0, 5, 10, and 50 mg/L） for 24 h. With a series of zymology and molecular biology methods, we found that HBCD induced dose-dependent oxidative stress on HBL-100 DNA. As revealed in q RT-PCR, activated prognostic factor ATM down-regulated tumor suppressor gene BRCA1 and prompted DNA repair genes h OGG1 and h MTH1 expression in lower concentrations of HBCD （〈 10 mg/L）. However, DNA repair were inhibited as well as cell proliferation rate by higher concentrations of HBCD （50 mg/L）. The results inferred that the genotoxicity of HBCD was dose-dependent and related to DNA repair pathway.
基金Supported by Key Project of Anhui Province Natural Science Foundation(KJ2008A085)Key Sci-tech Research Project of Anhui Province(08010302179)2008 NSFC General Project of China ( 30872253)~~
文摘[ Objective ] To develop a rapid efficient method for detecting mycoplasma contamination in cell cultures. [ Method] A pair of primers was designed according to two highly conserved nucleotide sequences of the 16S RNA from six kinds of mycoplasma that commonly contaminated cells. Then the mycoplasma contamination of 25 cell samples was defected by PCR and DNA fluorescence staining. EResultl When these cell samples were detected by DNA fluorescence staining, the positive rate and probable positive rate were respectively 24% and 16%. And when they were detected by PCR, the positive rate was 36%. [ Condusion] The PCR method is more sensitive and specific than the DNA fluorescence staining, and combining these two methods is the optimal way to detect mycoplasma contamination in cell cultures.
文摘The ＇double T-DNA＇ binary vector p13HSR which harbored two independent T-DNAs, containing hygromycin phosphotransferase gene （hpf） in one T-DNA region and three target genes （hLF, SB401, RZ10） in another T-DNA region, was used to generate selectable marker-free transgenic rice by Agrobacterium-mediated transformation. The regenerated plants with both the three target genes and the selectable marker gene hpt were selected for anther culture. RT-PCR analysis indicated that target genes were inserted in rice genomic DNA and successfully transcribed. It took only one year to obtain double haploid selectable marker-free transgenic plants containing the three target genes with co-transformation followed by anther culture technique, and the efficiency was 12.2%. It was also noted that one or two target genes derived from the binary vector were lost in some transgenic rice plants.
文摘目的:Siloranes是一种含硅的新型牙科复合树脂环氧乙烷单体,较传统的含碳的环氧乙烷单体具有更好的生物相容性。本研究的目的为用哺乳动物细胞培养检测姐妹染色体交换的方法评估Siloranes的基因毒性,并与传统的环氧乙烷比较。方法:研究所用的传统环氧乙烷为CyracureTM UVR6105。Silorane包括Ph-Sil,TET-Sil和二者50/50(w/w)的混合物(3M-ESPE)。用CHO细胞在有或无S9情况下检测姐妹染色体交换(SCE)。Bonferron post hoc统计分析检测有无显著性差异。结果:与溶剂对照相比,在有或无S9情况下,环氧乙烷UVR6105显著性地增加了培养CHO细胞的SCE数目。3种Silorane单体Ph-Sil,TET-Sil或Sil-Mix均未能增加SCE数目。结论:用哺乳动物细胞培养检测姐妹染色体交换的方法评估显示,3种新型Silorane单体Ph-Sil,TET-Sil或Sil-Mix均未检测出基因毒性效果,提示Silorane单体具有较低基因毒性,是适于研发牙科树脂的新型单体。
