探讨Mce4E蛋白在牛结核分枝杆菌致病机理中的作用。以Mce4E蛋白刺激肺泡巨噬细胞24和48h后,MTT检测分析表明,Mce4E蛋白对巨噬细胞的活性有显著的抑制作用(P<0.05);用重组表达的Mce4E蛋白、人分枝杆菌纯化蛋白衍生物(Purified Protein...探讨Mce4E蛋白在牛结核分枝杆菌致病机理中的作用。以Mce4E蛋白刺激肺泡巨噬细胞24和48h后,MTT检测分析表明,Mce4E蛋白对巨噬细胞的活性有显著的抑制作用(P<0.05);用重组表达的Mce4E蛋白、人分枝杆菌纯化蛋白衍生物(Purified Protein Derivative of M.tuberculosis,MtbPPD)、牛结核分枝杆菌纯化蛋白衍生物(Purified Protein Derivative of M.bovis,MbPPD)、卡介苗(Bacille Calmette Guerin,BCG)和刀豆蛋白A(con-canavalin A,ConA)分别刺激肺泡巨噬细胞48h后,real-time PCR方法检测显示,Mce4E蛋白能够促使牛肺泡巨噬细胞TNF-α和IL-6 mRNA的表达上调(P<0.05)、抑制肺泡巨噬细胞iNO的mRNA表达(P<0.05)而对IL-12表达没有影响(P>0.05)。Mce4蛋白能够促使肺泡巨噬细胞分泌炎性细胞因子,并对肺泡巨噬细胞有抑制作用,证实Mce4蛋白在分枝杆菌中具有重要的作用。展开更多
Objective This study aims to investigate and compare the toxic effects of four types of metal oxide(ZnO,TiO_(2),SiO_(2),and Al_(2)O_(3))nanoparticles with similar primary size(-20 nm)on human fetal lung fibroblasts(HF...Objective This study aims to investigate and compare the toxic effects of four types of metal oxide(ZnO,TiO_(2),SiO_(2),and Al_(2)O_(3))nanoparticles with similar primary size(-20 nm)on human fetal lung fibroblasts(HFL1)in vitro.Methods The HFL1 cells were exposed to the nanoparticles,and toxic effects were analyzed by using MTT assay,cellular morphology observation and Hoechst 33258 staining.Results The results show that the four types of metal oxide nanoparticles lead to cellular mitochondrial dysfunction,morphological modifications and apoptosis at the concentration range of 0.25-1.50 mg/mL and the toxic effects are obviously displayed in dose-dependent manner.ZnO is the most toxic nanomaterials followed by TiO_(2),SiO_(2),and Al_(2)O_(3)nanoparticles in a descending order.Conclusion The results highlight the differential cytotoxicity associated with exposure to ZnO,TiO_(2),SiO_(2),and Al_(2)O_(3)nanoparticles,and suggest an extreme attention to safety utilization of these nanomaterials.展开更多
CUEDC2(CUE domain containging protein 2)是近年来发现的一个功能尚未十分明确的蛋白质。实验室前期的研究表明,CUEDC2通过影响孕激素受体PR抑制乳腺癌细胞生长。此外,研究还发现CUEDC2通过招募PP1磷酸酶,促进IKK复合体的去磷酸化,抑...CUEDC2(CUE domain containging protein 2)是近年来发现的一个功能尚未十分明确的蛋白质。实验室前期的研究表明,CUEDC2通过影响孕激素受体PR抑制乳腺癌细胞生长。此外,研究还发现CUEDC2通过招募PP1磷酸酶,促进IKK复合体的去磷酸化,抑制NF-kB信号通路的激活。为了深入研究CUEDC2的功能,构建了CUED2可诱导表达载体(p617-neo-T-CUEDC2-I-tTA4),并通过逆转录病毒系统获得tet-off可诱导表达CUEDC2的稳定细胞系。该诱导表达细胞株的建立为CUEDC2功能基因的研究提供了必要的手段。展开更多
A quantitative protocol for the rapid analysis of Microcystis cells and colonies in lake sediment was developed using a modified flow cytometer, the CytoSense. For cell enumeration, diluted sediment samples containing...A quantitative protocol for the rapid analysis of Microcystis cells and colonies in lake sediment was developed using a modified flow cytometer, the CytoSense. For cell enumeration, diluted sediment samples containing Microcystis were processed with sonication to disintegrate colonies into single cells. An optimized procedure suggested that 5 mg dw （dry weight）/mL dilution combined with 200 W x 2 min sonication yielded the highest counting efficiency. Under the optimized determination conditions, the quantification limit of this protocol was 3.3x104 cells/g dw. For colony analysis, Microcystis were isolated from the sediment by filtration. Colony lengths measured by flow cytometry were similar to those measured by microscopy for the size range of one single cell to almost 400 ~tm in length. Moreover, the relationship between colony size and cell number was determined for three Microcystis species, including Microcystisflos-aquae, M. aeruginosa and M. wessenbergii. Regression formulas were used to calculate the cell numbers in different- sized colonies. The developed protocol was applied to field sediment samples from Lake Taihu. The results indicated the potential and applicability of flow cytometry as a tool for the rapid analysis of benthic Microcystis. This study provided a new capability for the high frequency monitoring of benthic overwintering and population dynamics of this bloom-forming cyanobacterium.展开更多
文摘探讨Mce4E蛋白在牛结核分枝杆菌致病机理中的作用。以Mce4E蛋白刺激肺泡巨噬细胞24和48h后,MTT检测分析表明,Mce4E蛋白对巨噬细胞的活性有显著的抑制作用(P<0.05);用重组表达的Mce4E蛋白、人分枝杆菌纯化蛋白衍生物(Purified Protein Derivative of M.tuberculosis,MtbPPD)、牛结核分枝杆菌纯化蛋白衍生物(Purified Protein Derivative of M.bovis,MbPPD)、卡介苗(Bacille Calmette Guerin,BCG)和刀豆蛋白A(con-canavalin A,ConA)分别刺激肺泡巨噬细胞48h后,real-time PCR方法检测显示,Mce4E蛋白能够促使牛肺泡巨噬细胞TNF-α和IL-6 mRNA的表达上调(P<0.05)、抑制肺泡巨噬细胞iNO的mRNA表达(P<0.05)而对IL-12表达没有影响(P>0.05)。Mce4蛋白能够促使肺泡巨噬细胞分泌炎性细胞因子,并对肺泡巨噬细胞有抑制作用,证实Mce4蛋白在分枝杆菌中具有重要的作用。
基金supported by grants from the National Basic Research Program of China(2011CB933404)the Science Foundation of Jiangsu Key Laboratory for Biomaterials and Devices(2010LBMD05)the Science Foundation of Southeast University(XJ2008335)
文摘Objective This study aims to investigate and compare the toxic effects of four types of metal oxide(ZnO,TiO_(2),SiO_(2),and Al_(2)O_(3))nanoparticles with similar primary size(-20 nm)on human fetal lung fibroblasts(HFL1)in vitro.Methods The HFL1 cells were exposed to the nanoparticles,and toxic effects were analyzed by using MTT assay,cellular morphology observation and Hoechst 33258 staining.Results The results show that the four types of metal oxide nanoparticles lead to cellular mitochondrial dysfunction,morphological modifications and apoptosis at the concentration range of 0.25-1.50 mg/mL and the toxic effects are obviously displayed in dose-dependent manner.ZnO is the most toxic nanomaterials followed by TiO_(2),SiO_(2),and Al_(2)O_(3)nanoparticles in a descending order.Conclusion The results highlight the differential cytotoxicity associated with exposure to ZnO,TiO_(2),SiO_(2),and Al_(2)O_(3)nanoparticles,and suggest an extreme attention to safety utilization of these nanomaterials.
文摘CUEDC2(CUE domain containging protein 2)是近年来发现的一个功能尚未十分明确的蛋白质。实验室前期的研究表明,CUEDC2通过影响孕激素受体PR抑制乳腺癌细胞生长。此外,研究还发现CUEDC2通过招募PP1磷酸酶,促进IKK复合体的去磷酸化,抑制NF-kB信号通路的激活。为了深入研究CUEDC2的功能,构建了CUED2可诱导表达载体(p617-neo-T-CUEDC2-I-tTA4),并通过逆转录病毒系统获得tet-off可诱导表达CUEDC2的稳定细胞系。该诱导表达细胞株的建立为CUEDC2功能基因的研究提供了必要的手段。
基金supported by the National Basic Research Program (973) of China (No. 2008CB418006)the National Special Program of Water Environment (No.2009ZX07106-001-002)+1 种基金the National Natural Science Foundation of China (No. 31070355)the National Major Science and Technology Program for Water Pollution Control and Treatment (No. 2009ZX07101-013)
文摘A quantitative protocol for the rapid analysis of Microcystis cells and colonies in lake sediment was developed using a modified flow cytometer, the CytoSense. For cell enumeration, diluted sediment samples containing Microcystis were processed with sonication to disintegrate colonies into single cells. An optimized procedure suggested that 5 mg dw （dry weight）/mL dilution combined with 200 W x 2 min sonication yielded the highest counting efficiency. Under the optimized determination conditions, the quantification limit of this protocol was 3.3x104 cells/g dw. For colony analysis, Microcystis were isolated from the sediment by filtration. Colony lengths measured by flow cytometry were similar to those measured by microscopy for the size range of one single cell to almost 400 ~tm in length. Moreover, the relationship between colony size and cell number was determined for three Microcystis species, including Microcystisflos-aquae, M. aeruginosa and M. wessenbergii. Regression formulas were used to calculate the cell numbers in different- sized colonies. The developed protocol was applied to field sediment samples from Lake Taihu. The results indicated the potential and applicability of flow cytometry as a tool for the rapid analysis of benthic Microcystis. This study provided a new capability for the high frequency monitoring of benthic overwintering and population dynamics of this bloom-forming cyanobacterium.
