Objective To study the regulatory roles of SIRT1 on EZH2 expression and the further ef-fects on EZH2's repression of target gene expression. Methods The stable SIRT1 RNAi and Control RNAi HeLa cells were establish...Objective To study the regulatory roles of SIRT1 on EZH2 expression and the further ef-fects on EZH2's repression of target gene expression. Methods The stable SIRT1 RNAi and Control RNAi HeLa cells were established by in-fection with retroviruses expressing shSIRT1 and shLuc respectively followed by puromycin selection. EZH2 protein level was detected by Western blot in either whole cell lysate or the fractional cell extract. Reverse transcription-polymerase chain reaction was performed to detect the mRNA level of EZH2. Cycloheximide was used to treat SIRT1 RNAi and Control RNAi cells for protein stability assay. Chromatin immunoprecipitation (ChIP) assay was applied to measure enrichment of SIRT1, EZH2, and trimethylated H3K27 (H3K27me3) at SATB1 promoter in SIRT1 RNAi and Control RNAi cells. Results Western blot results showed that EZH2 protein level increased upon SIRT1 de-pletion. Fractional extraction results showed unchanged cytoplasmic fraction and increased chromatin fraction of EZH2 protein in SIRT1 RNAi cells. The mRNA level of EZH2 was not affected by knockdown of SIRT1. SIRT1 recruitment was not detected at the promoter region of EZH2 gene locus. The protein stability assay showed that the protein stability of EZH2 increases upon SIRT1 knockdown. Upon SIRT1 depletion, EZH2 and H3K27me3 recruitment at SATB1 promoter increases and the mRNA level of SATB1 decreases. Conclusions Depletion of SIRT1 increases the protein stability of EZH2. The regulation of EZH2 protein level by SIRT1 affects the repressive effects of EZH2 on the target gene expres-sion.展开更多
It is necessary to treat wastewater in a more environment-friendly fashion.Vegetated submerge bed constructed wetlands (CWs) has become the focus of the research in this field. In this paper, the overall capacity of v...It is necessary to treat wastewater in a more environment-friendly fashion.Vegetated submerge bed constructed wetlands (CWs) has become the focus of the research in this field. In this paper, the overall capacity of vertical subsurface flow CWs in the removal of pollutants from municipal wastewater effluent and the effects of the depth of vegetated submerged bed on the removal efficiency were studied. The results showed that the COD, NH+4-N and TN could be removed effectively in the vertical subsurface flow CWs and the best efficiency was obtained from the 10 cm run. However, the TP removal was not effective in all vertical CWs. A shallow depth of vegetated submerged bed may avoid the risk of substrate plugging effectively, and may keep a stable operation of CWs in long term. Decreasing the depth of vegetated submerged bed could create a sufficient aerobic circumstance in which the concentration of DO (dissolved oxygen) in bed was super saturation.展开更多
Reference materials for quantitative determination of regulated heavy metals, such as Pb and Cd in electronic components, were designed and investigated in terms of stability and homogeneity. Reference materials with ...Reference materials for quantitative determination of regulated heavy metals, such as Pb and Cd in electronic components, were designed and investigated in terms of stability and homogeneity. Reference materials with two concentration levels of heavy metals were prepared by spiking Pb and Cd compounds to epoxy molding compounds made by mixing silica powders and epoxy resin. The concentration changes of the reference materials during stability test for 1 a were not observed. In the homogeneity assessment, the as-prepared reference materials were studied by using three different analytical tools, inductively coupled plasma atomic emission spectrometry (ICP-AES), X-ray fluoroescence spectrometry (XRF) and laser ablation ICP mass. The results show different homogeneities by the characteristics of analytical tools and the materials.展开更多
基金Supported by National Natural Science Foundation of China (30721063)National Basic Research Program of China (973 Program) (2005CB522402, 2006CB910403)+1 种基金National Laboratory of Medical Molecular Biology grant (2060204)Beijing municipal government grant (YB20081002301)
文摘Objective To study the regulatory roles of SIRT1 on EZH2 expression and the further ef-fects on EZH2's repression of target gene expression. Methods The stable SIRT1 RNAi and Control RNAi HeLa cells were established by in-fection with retroviruses expressing shSIRT1 and shLuc respectively followed by puromycin selection. EZH2 protein level was detected by Western blot in either whole cell lysate or the fractional cell extract. Reverse transcription-polymerase chain reaction was performed to detect the mRNA level of EZH2. Cycloheximide was used to treat SIRT1 RNAi and Control RNAi cells for protein stability assay. Chromatin immunoprecipitation (ChIP) assay was applied to measure enrichment of SIRT1, EZH2, and trimethylated H3K27 (H3K27me3) at SATB1 promoter in SIRT1 RNAi and Control RNAi cells. Results Western blot results showed that EZH2 protein level increased upon SIRT1 de-pletion. Fractional extraction results showed unchanged cytoplasmic fraction and increased chromatin fraction of EZH2 protein in SIRT1 RNAi cells. The mRNA level of EZH2 was not affected by knockdown of SIRT1. SIRT1 recruitment was not detected at the promoter region of EZH2 gene locus. The protein stability assay showed that the protein stability of EZH2 increases upon SIRT1 knockdown. Upon SIRT1 depletion, EZH2 and H3K27me3 recruitment at SATB1 promoter increases and the mRNA level of SATB1 decreases. Conclusions Depletion of SIRT1 increases the protein stability of EZH2. The regulation of EZH2 protein level by SIRT1 affects the repressive effects of EZH2 on the target gene expres-sion.
基金supported by the Scientific Research Foundation of the State Human Resource Ministry for Returned Chinese Scholars, China (Grant No.SLZ2008012)Natural Science Basic Research Plan in Shaanxi Province of China (Grant No. SJ08-ZT07)+1 种基金the Research Program of the Education Department of Shannxi Province of China (Grant No. 08JK327)Program for Changjiang Scholars and Innovative Research Team in University(PCSIRT) (Grant No. IRT0853)
文摘It is necessary to treat wastewater in a more environment-friendly fashion.Vegetated submerge bed constructed wetlands (CWs) has become the focus of the research in this field. In this paper, the overall capacity of vertical subsurface flow CWs in the removal of pollutants from municipal wastewater effluent and the effects of the depth of vegetated submerged bed on the removal efficiency were studied. The results showed that the COD, NH+4-N and TN could be removed effectively in the vertical subsurface flow CWs and the best efficiency was obtained from the 10 cm run. However, the TP removal was not effective in all vertical CWs. A shallow depth of vegetated submerged bed may avoid the risk of substrate plugging effectively, and may keep a stable operation of CWs in long term. Decreasing the depth of vegetated submerged bed could create a sufficient aerobic circumstance in which the concentration of DO (dissolved oxygen) in bed was super saturation.
基金Project(2010-0008-276) supported by NCRC (National Core Research Center) program through the National Research Foundation of Korea funded by the Ministry of Education, Science and Technology
文摘Reference materials for quantitative determination of regulated heavy metals, such as Pb and Cd in electronic components, were designed and investigated in terms of stability and homogeneity. Reference materials with two concentration levels of heavy metals were prepared by spiking Pb and Cd compounds to epoxy molding compounds made by mixing silica powders and epoxy resin. The concentration changes of the reference materials during stability test for 1 a were not observed. In the homogeneity assessment, the as-prepared reference materials were studied by using three different analytical tools, inductively coupled plasma atomic emission spectrometry (ICP-AES), X-ray fluoroescence spectrometry (XRF) and laser ablation ICP mass. The results show different homogeneities by the characteristics of analytical tools and the materials.
