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An efficient method of sorting liver stem cells by using immuno.magnetic microbeads

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World Journal of Gastroenterology 2006年第19期12卷 3050-3054页

作者： Yu-Fei He Yin-Kun Liu Dong-Mei Gao Jun Chen Peng-Yuan Yang Institute Zhongshan Hospital Fudan University Shanghai 200032 China Center of Proteomics BioMedical Institutes Fudan University Shanghai 200032 China

AIM： To develop a method to isolate liver stem cells fast and efficiently. METHODS： Fetal mouse liver cells were characterized by cell surface antigens （c-Kit and CD45/TER119） using flow cytometry. The candidate liver stem cells were sorted by using immuno.magnetic microbeads and identified by -lone-forming culture, RT-PCR and immuno.luorescence says. RESULTS： The c-Kit-（CD45/TER119）- cell population with 97.9% of purity were purified by immuno.magnetic microbeads at one time. The yield of this separation was about 6% of the total sorting cells and the cell viability was above 98%. When cultured in vitro these cells had high clone-forming and self-renewing ability and expressed markers of hepatocytes and bile duct cells. Functionally mature hepatocytes were observed after 21 d of culture. CONCLUSION： This method offers an excellent tool for the enrichment of liver stem cells with high purity and viability, which could be used for further studies. It is fast, efficient, simple and no. expensive.

关键词： Liver stem cells Immuno.magnetic microbe.ads Sorting Flow cytometry

Proteomic dissection of biological pathways/processes through profiling protein-protein interaction networks

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science China Chemistry 2010年第4期53卷 737-746页

作者： CHEN Xian Institutes for Biomedical sciences, Fudan University, shanghai 200433, China Department of Biochemistry & Biophysics, School of Medicine, University of no.th Carolina-Chapel Hill, USA Institutes for Biomedical sciences Fudan University Shanghai China Department of Biochemistry & Biophysics School of Medicine University of North Carolina-Chapel Hill Chapel Hill USA

Cellular functions, either under the no.mal or pathological conditions or under different stresses, are the results of the coordinated action of multiple proteins interacting in macromolecular complexes or assemblies. The precise determination of the specific composition of protein complexes, especially using scalable and high-throughput methods, represents a systematic approach toward revealing particular cellular biological functions. In this regard, the direct profiling protein-protein interactions (PPIs) represent an efficient way to dissect functional pathways for revealing no.el protein functions. In this review, we illustrate the techno.ogical evolution for the large-scale and precise identification of PPIs toward higher physiologically relevant accuracy. These techniques aim at improving the efficiency of complex pull-down, the signal specificity and accuracy in distinguishing specific PPIs, and the accuracy of identifying physiological relevant PPIs. A newly developed streamline proteomic approach for mapping the binary relationship of PPIs in a protein complex is introduced.

关键词： Chen Proteomic dissection of biological pathways/processes through profiling protein-protein interaction networks

Nitrification characteristics of nitrobacteria immobilized in waterborne polyurethane in wastewater of corn-based ethano. fuel production

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Journal of Environmental sciences 2012年第6期24卷 999-1005页

作者： Yamei Dong Zhenjia Zhang Yongwei Jin Jian Lut Xuehang Cheng Jun Li Yan-yan Deng Ya-nan Feng Dongning Chen School of Environmental science and Engineering Shanghai Jiao Tong UniversityShanghai 200240China National Engineering Research Center for Nano.0} Shanghai 200241China Indian River Research and Education Center University of FloridaFort PierceFL 34945USA Division of Environmental science and Engineering Yamaguchi UniversityTokiwadai 2-16-1Yamaguchi 755-8611Japan School of Municipal and Environmental Engineering Shengyang Jianzhu UniversityShenyang 110168China

A technology to achieve stable and high ammonia nitrogen removal rates for corn distillery wastewater （ethano. fuel production） treatment has been *** characteristics of nitrifying bacteria entrapped in a waterborne polyurethane （WPU） gel carrier were evaluated after *** the acclimation period,nitrification rates of WPU-immobilized nitrobacteria were monitored and polymerase chain reaction （PCR） was also carried out to investigate the change in ammonium-oxidizing *** results showed that the pellet nitrification rates increased from 21 to 228 mg-N/（L-pellet·hr） and the quantity of the ammonia oxidation bacteria increased substantially during the acclimation.A continuous ammonia removal experiment with the anaerobic pond effluent of a distillery wastewater system was conducted with immobilized nitrifying bacteria for 30 days using an 80 L airlift reactor with pellets at a fill ratio of 15% （V/V）.Under the conditions of 75 mg/L influent ammonia,hydraulic retention time （HRT） of 3.7-5.6 hr,and dissolved oxygen （DO） of 4 mg/L,the effluent ammonia concentration was lower than 10 mg/L and the ammonia removal efficiency was 90%.While the highest ammonia removal rate,162 mg-N/（L-pellet·hr）,was observed when the HRT was 1.3 hr.

关键词： immobilized nitrobacteria corn distillery wastewater ethano. fuel production nitrification rate waterborne polyurethane gel

"Small amount for multiple times" of H_(2)O_(2) feeding way in MoS_(2)-Fex heterogeneous fenton for enhancing sulfadiazine degradation

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Chinese Chemical Letters 2022年第3期33卷 1365-1372页

作者： Zhuan Chen Cheng Lian Kai Huang Jiahui Ji Qingyun Yan Jinlong Zhang Mingyang Xing Key Laboratory for Advanced Materials and Joint International Research Laboratory of Precision Chemistry and Molecular Engineering Feringa Nobel Prize Scientist Joint Research CenterFrontiers Science Center for Materiobiology and Dynamic ChemistryInstitute of Fine ChemicalsSchool of Chemistry and Molecular EngineeringEast China University of Science and TechnologyShanghai 200237China shanghai Engineering Research Center for Multi-media Environmental Catalysis and Resource Utilization East China University of Science and TechnologyShanghai 200237China

In recent years, MoS_(2) catalyzed/cocatalyzed Fenton/Fenton-like systems have attracted wide attention in the field of pollution control, but there are few studies on the effect of H_(2)O_(2) feeding way on the whole Fenton process. Here, we report a new type of composite catalyst (MoS_(2)-Fe_(x)) prepared in a simple way with highly dispersed iron to provide more active sites. MoS_(2)-Fe_(x) was proved to possess selectivity for singlet oxygen (^(1)O_(2)) in effectively degrading sulfadiazine with a wide pH adaptability (4.0~10.0). Importantly, the mechanism of the interaction between H_(2)O_(2) and MoS_(2) on the Fenton reaction activity was revealed through the combination of experiment and density functional theory (DFT) calculations. Compared to the traditional “a large amount for one time” feeding way of H_(2)O_(2), the “small amount for multiple times” of H_(2)O_(2) feeding way can increase the degradation rate of sulfadiazine from 36.9% to 91.1% in the MoS_(2)-Fe_(x) heterogeneous Fenton system. It is demonstrated that the “small amount for multiple times” of H_(2)O_(2) feeding way can reduce the side reaction of decomposition of H_(2)O_(2) by MoS_(2) and effectively improve the utilization rate of H_(2)O_(2) and the stability of MoS_(2)-Fe_(x). Compared with Fe_(2)O_(3)^(-)based Fenton system, MoS_(2)-Fe_(x) can significantly save the amount of H_(2)O_(2). Compared with nano.iron powder, the formation of iron sludge in MoS_(2)-Fe_(x) system was significantly reduced. Furthermore, long-term degradation test showed that the MoS_(2)-Fe75/H_(2)O_(2) system could maintain the effectiveness of degrading organic pollutants for 10 days (or even longer). This study has a guiding significance for the large-scale treatment of industrial wastewater by improved Fenton technology in the future.

关键词： Fenton reaction Co-catalytic Molybdenum sulfide Hydrogen peroxide Singlet oxygen

PCR-based approaches for identification of multi-copy transgene integration sites in mouse geno.e

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Chinese science Bulletin 2006年第18期51卷 2231-2235页

作者： ZHAO Xudong DANG Suying LIANG Bin LEI Xia CHEN Zheng WANG Long YAN Lanzhen SUN Hantang FU Jiliang FEI Jian WANG Zhugang shanghai Research Center for Model Organisms Shanghai 201203 China Laboratory of Genetic Engineering Department of Medical Genetics Institute of Health Science jointly-funded by Shanghai Institutes for Biological Sciences of Chinese Academy of Sciences and Shanghai Second Medical University Shanghai 200025 China State Key Laboratory of Medical Geno.ics Rui-Jin Hospital Affiliated to Shanghai Second Medical University Shanghai 200025 China

Generation of transgenic mice by DNA microinjection has become one of the most important techno.ogies in studying gene function in vivo. Ran- dom integration of transgene often results in inser- tional mutation which may complicate pheno.ype analysis of transgenic mice and/or create a good opportunity to study the function of endogeno.s gene. However,the utilization of these potentially valuable resources is hampered due to lack of efficient ap- proaches for rapid identification of multi-copy trans- gene integration sites. Here we propose two PCR-based approaches to identifying trans- gene/geno.e junction sequences. The efficiency of these two strategies was tested in 9 independent transgenic mouse lines. Among them,the transgene in 8 mouse lines was clearly localized to certain chromosome regions. These rapid and efficient ap- proaches will greatly facilitate the study of insertional mutation due to transgene integration.

关键词：转基因结合部位插入突变 PCR

The predictive value of chromosome 8p deletion for metastasis of hepatocellular carcino.a:a summary of works in 10 years

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Frontiers of Medicine 2008年第3期2卷 211-215页

作者： Lunxiu QIN Zhaoyou TANG Xinyuan GUAN Qinghai YE Huliang JIA Ning REN Liver Cancer Institute&Zhongshan Hospital Fudan UniversityShanghai 200032China

Hepatocellular carcino.a(HCC)represents an extremely poor progno.tic cancer,which is mainly due to the high frequency of metastasis/recurrence after surgical *** the molecular mechanisms involved in HCC metastasis could be helpful in the pre-diction and early diagno.is of HCC recurrence and could also provide new therapeutic targets for HCC *** the recent decade,we analyzed the geno.ic aberrations of the clinical specimens,as well as the metastatic models and cell lines of human HCC to identify the genetic mar-kers related to HCC metastasis and to verify their clinical values in the prediction and control of metastasis of *** the comparative geno.ic hybridization(CGH)technique,we compared the differences of chromosomal aberrations between primary HCC tumors and their matched metastatic lesions,and found that chromosome 8p deletions might contribute to HCC *** no.el finding was further confirmed by comparison between nude mice models of HCC with different meta-static *** the more sensitive geno.e-wide microsatellite analysis,8p deletion was defined to 8p23.3 and 8p11.2,which are two likely regions harboring meta-stasis-related genes of ***‘8p-specific’microar-rays,two no.el metastatic suppressors(HTPAP and MRSA)were identified,and were proven to suppress in vitro invasion and in vivo metastasis of *** studies indicate that 8p deletion detected in HCC or cir-culating plasma DNA of patients is a useful predictor for metastatic recurrence and progno.is,even for patients with early stage *** no.el findings are regarded as important advances in the study of the molecular mechanisms of HCC metastasis,which provide no. only a holistic view on the molecular cytogenetic bases of HCC metastasis,but also candidate regions for further study to identify metastatic suppressor genes.

关键词： hepatocellular carcino.a metastasis chromosomes,human,pair 8 genes,tumor suppressor prediction progno.is