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Development of a quantitative,cell-based,high-content screening assay for epidermal growth factor receptor modulators

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Acta Pharmacologica Sinica 2007年第10期28卷 1698-1704页

作者： Jue WANG~2.Xin XIE~(2.3)~2.The National Center for Drug Screening,shanghai Institute of Materia Medica,Chinese Academy of Sciences graduate {2. of Chinese Academyof Sciences shanghai 2.12.3,china The National Center for Drug Screening Shanghai Institute of Materia Medica Chinese Academy of Sciences graduate school of Chinese Academy of Sciences Shanghai China

Aim:To develop a robust,cell-based,high-content screening （HCS） assay basedon receptor internalization for the identification of novel modulators of the epi-dermal growth factor receptor （EGFR）.Methods:Agonist-induced receptorinternalization is part of the signaling cascade of ***-taggedepidermal growth factor （EGF） was used to visualize the internalized receptor-ligand *** fluorescent intracellular spots were detected and mea-sured with an ArrayScan HCS *** that can competitively bind toEGFR or interfere with EGFR internalization process would result in a reducednumber and intensity of intracellular fluorescent *** assay was validated,optimized,and applied to a large-scale screening of a library containing 48 000synthetic ***:The competition between fluorescent EGF andunlabeled EGF reveals the IC50of unlabeled EGF is approximately 0.2.nmol/L,which is comparable with other published *** compounds with arelatively high degree of interference with EGFR internalization were *** of the compounds was proven to be agonist of the EGFR since it inducedphosphorylation of the receptor and extracellular signal-regulated protein ki-nase （ERK）.Conclusion:This automated,objective,and easy-to-use assay pro-vided abundant information,quantitative results,and demonstrated the potentialuse of HCS methods in searching membrane receptor modulators.

关键词： high-content screening epidermal growth factor receptor receptor internalization competitive binding

Characteristics and geological significance of olivine xenocrysts in Cenozoic volcanic rocks from western Qinling

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Progress in Natural Science:Materials International 2006年第12期16卷 1300-1306页

作者： SU Benxun~ 1,2.**, ZHANG Hongfu~1, XIAO Yan~ 1,2.and ZHAO Xinmiao~ 1,2.(1. State Key Laboratory of Lithospheric Evolution, Institute of Geology and Geophysics, Chinese Academy of Sciences, Beijing 10002., china 2. graduate {2. of Chinese Academy of Sciences, Beijing 100039, china) State Key Laboratory of Lithospheric Evolution Institute of Geology and Geophysics Chinese Academy of Sciences Beijing 100029 China graduate school of Chinese Academy of Sciences Beijing 100039 China

Cenozoic volcanic rocks from the Haoti, Dangchang County of the western Qinling Mountains, contain a few clearly-zoned olivines. These olivines are relatively big in grain sizes and usually have cracks or broken features. Their cores have similar compositions （Mg#=90.4-91.0） to those for the peridotitic xenoliths entrained in host volcanic rocks and their rims are close to the compositions of olivine phenocrysts （Mg#=85.5-81.9）. The CaO contents in these zoned olivines are lower than 0.1%. These features demonstrate that the clearly-zoned olivines are xenocrysts and disaggregated from mantle peridotites. The zoned texture was the result of the interaction between the olivine and host magma. Available data show that the volcanic rocks would have been derived from the mantle source metasomatized by subducted hydrathermally-altered oceanic crust. The formation of these Cenozoic volcanic rocks was perhaps related to the rapid uplift of the Tibetan Plateau.

关键词： Western Qinling Dangchang Haoti Cenozoic volcanic rocks olivine xenocrysts zoned texture.

Dynamic learning rates algorithm for BPNN to forecast time series of dam security

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Progress in Natural Science:Materials International 2007年第12期17卷 1527-1530页

作者： Hu Daiping1, Wu Ruiming1, Lei Aizhong1 and Wang Qi2.1. Antai College of Economics & Management, shanghai Jiaotong University, shanghai 2.0052. {2. 2. graduate school, Lanzhou University, Lanzhou 730000, {2.) Antai College of Economics and Management Shanghai Jiaotong University Shanghai 200052 China graduate school Lanzhou University Lanzhou 730000 China

Time series data of dam security have a large number of observed values and should be forecasted accurately in time. Neural networks have the powerful approach ablilities of arbitrary functions and have been broadly utilized in many domains. In this paper, a dynamic learning rate training algorithm of back-propagation neural networks for time series forecasting is proposed and the networks with this algorithm are built to forecast time series of dam security. The application results demonostrate the efficiency of modelling and the effictiveness of forecasting.

关键词： neural network dynamic learning rate forecasting dam security.

Bio-regeneration of p-complexation desulfurization adsorbents

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Science china Chemistry 2005年第6期48卷 538-544页

作者： LI Wangliang1,2. XING Jianmin1, XIONG Xiaochao1,2. SHAN Guobin1,2.& LIU Huizhou1 1. Laboratory of Separation Science and Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100080, china 2. graduate {2. of the Chinese Academy of Sciences,Beijing 100039, china Laboratory of Separation Science and Engineering Institute of Process Engineering Chinese Academy of Sciences Beijing China graduate school of the Chinese Academy of Sciences Beijing China

The coupling of adsorption desulfurization and biodesulfurization is a new approach to produce clean fuels. Sulfur compounds are firstly adsorbed on adsorbents, and then the ad-sorbents are regenerated by microbial conversion. p-Complexation adsorbent, Cu(I)-Y, was ob-tained by ion exchanging Y-type zeolite with Cu2. and then by auto-reduction in helium at 450℃ for 3 h. Dibenzothiophene (DBT) was used as a model compound. The effects of cell concentra-tion, volume of oil phase, the ratio of aqueous phase to adsorbent on DBT desorption by a bac-terium were studied. The amounts of DBT desorbed and 2.HBP produced can be apparently increased with addition of n-octane. BDS activity can be improved by increasing cell concentra-tion and the ratio of water-to-adsorbent. 89% of DBT desorbed from the adsorbents can be converted to 2.HBP within 6 h and almost 100% within 2. h, when the volume ratio of oil-to-water was 1/5 mL/mL, the cell concentration was 60 g·L?1, and the ratio of adsorbent-to-oil was 0.03 g·mL?1. The amount of 2.HBP produced was strongly dependent on the volume ratio of oil-to- water, cell concentration and amount of adsorbent. Adsorption capacity of the regenerated ad-sorbent is 95% that of the fresh one after being desorbed with Pseudomonas delafieldii R-8, washed with n-octane, dried at 100℃ for 2. h and auto-reduced in He.

关键词： adsorption desulfurization p-complexation adsorbents bio-regeneration Pseudomonas delafieldii desorption.

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Prevention of lipopolysaccharide-induced injury by 3,5-dicaffeoylquinic acid in endothelial cells

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Acta Pharmacologica Sinica 2007年第8期28卷 1143-1148页

作者： Ruo-peng ZHA~(1,2. Wei XU~(1,2. Wen-yi WANG~(1,2. Li DONG~1 Yi-ping WANG~(1,3) ~1State Key Laboratory of Drug Research,shanghai Institute of Materia Medica,shanghai Institutes for Biological Sciences,Chinese Academy of Seiences,shanghai 2.12.3,china ~2.4} school of the Chinese Academy of Sciences,shanghai 2.12.3,china State Key Laboratory of Drug Research Shanghai Institute of Materia Medica Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China graduate school of the Chinese Academy of Sciences Shanghai China

Aim:To investigate the effect of 3,5-dicaffeoylquinic acid（3,5-diCQA）onlipopolysaccharide（LPS）-induced injury in human dermal microvascular endothe-lial cells（HMEC-I）.Methods:The anti-oxidant effect was detected using themalondialdehyde（MDA）assay in a rat liver microsome model of lipid *** viability was analyzed using the 3-（4,5-dimethylthiazol-2.yl）-2.5diphenyltetrazolium bromide *** lipid peroxide injury was measured bylactate dehydrogenase（LDH）*** cells were detected by flowcytometry,and confirmed by DNA fragmentation ***-3 activitywas measured using a specific assay *** level of intracellular reactive oxygenspecies（ROS）was determined by flow cytometry with a 2.7-dichlorodihydro-fluorescein diacetate fluorescence ***:The exposure of microsomesto ascorbate-Fe2.resulted in lipoperoxidation according to an increase in the levelof *** formation decreased in a dose-dependent manner on treatmentwith 5,10,or 50μmol/L 3,*** with LPS for 16 h resulted in a 60%decrease in cell viability and an increase in LDH release from 47.6% to 61.5%.DNA laddering was observed by agarose gel *** level ofapoptotic cells peaked at 2.% after treatment with LPS for 12.*** treat-ment with LPS for 12.h,intracellular ROS and caspase-3 activity ***-treatment with 3,5-diCQA at 5,10,or 50μmol/L for 1 h attenuated LPS-mediatedendothelial cell *** anti-apoptotic action of 3,5-diCQA was partiallydependent on its capacity for anti-oxidation and the suppression of ***:3,5-diCQA displays anti-oxidative and anti-apoptotic activ-ity in HMEC-1 due to scavenging of intracellular ROS induced by LPS,and thesuppression of caspase-3 activity.

关键词： 3 5-dicaffeoylquinic acid lipopolysaccharide human dermal microvascular endo-thelial cells reactive oxygen species apoptosis caspase-3

Apoptosis initiated by carbon tetrachloride in mitochondria of rat primary cultured hepatocytes

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Acta Pharmacologica Sinica 2005年第8期26卷 969-975页

作者： Yan CAI~(1,2., Li-kun GONG~(1,2., Xin-ming QI~(1,2., Xiang-hong LI~1, Jin REN~(1,3) ~1State Key Laboratory of Drug Researeh, shanghai Institute of Materia Medica, shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, shanghai 2.12.3, china ~2.4} school of the Chinese Academy of Sciences, shanghai 2.12.3, china State Key Laboratory of Drug Research Shanghai Institute of Materia Medica Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China graduate school of the Chinese Academy of Sciences Shanghai China

Aim: To investigate the mitochondria-initiated apoptosis pathway involved in Carbon tetrachloride （CCl4） hepatotoxicity in vitro. Methods: Several cytotoxic-ity endpoints, including WST-8 metabolism, lactate dehydrogenase leakage and morphological changes, were examined. The 5,5'-dithio-bis（2.nitrobenzoic acid） reaction was used to measure reduced glutathione level, and the malondialdehyde level was determined using the thiobarbituric acid assay. The release of cyto-chrome c and Bcl-XLwas detected by Western blot. Caspase-3 activity was mea sured using the fluorogenic substrate Ac-DEVD-AMC. DNA fragmentation wasused to evaluate cell apoptosis. Results: A time-and dose-dependent decrease in cellular glutathione content was observed, along with a concomitant increase in malondialdehyde levels following the application of CCl4. Caspase 3 activity was stimulated at all doses of CCl4, with the most significant activation at 3 mmol/L. Cytochrome c was released obviously after CCl4treatment. A time-dependent decrease in BcI-XLexpression was observed. DNA fragmentation results revealed apoptosis and necrosis following CCl4treatment. Conclusion: Oxidative damage is one of the essential mechanisms of CCl4hepatotoxicity, which triggers apoptosis via the mitochondria-initiated pathway.

关键词： carbon tetrachloride hepatocytes mitochondria caspase 3 cytochrome c

Altered expression of cytochrome P450 and possible correlation with preneoplastic changes in early stage of rat hepatocarcinogenesis

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Acta Pharmacologica Sinica 2005年第6期26卷 737-744页

作者： Lin-lin LIU~(2.3), Li-kun GONG~(2.3), Xin-ming QI~(2.3),Yan CAI~(2.3), Hui WANG~2. Xiong-fei WU~(2.3),Ying XIAO~(2.3), Jin REN~(2.4) ~2.State Key Laboratory of Drug Research, shanghai Institute of Materia Medica, shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, shanghai 2.12.3, china ~3 graduate school of the Chinese Academy of Sciences, shanghai 2.12.3, china State Key Laboratory of Drug Research Shanghai Institute of Materia Medica Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China graduate school of the Chinese Academy of Sciences Shanghai China

Aim: Correlation of cytochrome P450 (CYPs) with preneoplastic changes in the early stage of hepatocarcinogenesis is still unclear. To detect the expression of carcinogen-metabolizing related microsomal P450 enzymes, namely the CYP1 Al, CYP1A2. CYP2.1/2. CYP2.1, and CYP3A, we performed the medium-term bioas-say of Ito's model in Sprague-Dawley rats. Methods: The amount and activity of CYP were assessed by biochemical and immunohistochemical methods in week 8. The correlation between CYP expression and microsomal oxidative stress was investigated by comparing the generation of microsomal lipid peroxidation in the presence or absence of specific CYP inhibitor. Results: In the DEN-2.AAF and 2.AAF alone groups, the expression of CYP1 Al and CYP2.1 were up-regulated and the expression of CYP2.1/2.and CYP1A2.were quite the contrary. Strong staining of CYP2.1 and CYP2.1/2.was found around the centrolobular vein and weak staining in the altered hepatic foci revealed by immunohistochemical procedure. There was no significant change in the activity of CYP3A among the 4 groups. Altered hepatic tissue bore more microsomal NADPH (nicotinamide adenine dimucleotide phosphate,reduced form)-dependent lipid peroxidation than normal tissue. And the difference among the 4 groups disappeared when CYP2.1 was inhibited. More microsomal lipid peroxidation was generated when incubated with CYP1A inhibitor a-naphthoflavone. Conclusion: CYP altered their expression levels and these alterations can play important roles in the alteration of cell redox status of preneoplastic tissue in the early stage of hepatocarcinogenesis.

关键词： cytochrome P450 preneoplastic changes hepatocarcinogensis

Novel Tumor-associated Antigen of Hepatocellular Carcinoma Defined by Monoclonal Antibody E4-65

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Acta Biochimica et Biophysica Sinica 2007年第5期39卷 359-365页

作者： Ke ZOU~(1,2., Jihang JU~(1,2., and Hong XIE~~(1*)1 Institute of Biochemist13, and Cell Biology, shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, shanghai 2.0031, {2.. 2.graduate school of the Chinese Academy of Sciences Beijing 100049, {2. Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of SciencesShanghai 200031 China graduate school of the Chinese Academy of SciencesBeijing 100049 China

A monoclonal antibody, E4-65, produced by immunizing mice with SMMC-772. cells, a human hepatocellular carcinoma (HCC) cell line, was used to identify and characterize an unreported HCC-associated antigen. Indirect immunofluorescence studies showed that E4-65 antibody reacted with five out of eight HCC cell lines, but not with 10 non-HCC tumor cell lines or a normal liver cell line. Using immunohistochemical examination, E4-65 antigen was detected on the cell membranes and in the cytoplasm of human liver tumor tissues, but was not found in most other tumors, or normal adult or fetal tissues, except for a weakly positive reaction in tissues of the digestive system. Western blot analysis showed that E4-65 antibody bound to a 45 kDa protein in the human HCC cell line and tissue lysates. Enzyme treatment and lectin blotting did not detect the carbohydrate chain in E4-65 antigen. This HCC-associated protein represents a potentially useful target for diagnoses and immunotherapy of human HCC.

关键词： hepatocellular carcinoma immunohistochemistry monoclonal antibody neoplasm antigen

Paleoceanographic evolution recorded in the northern South china Sea since 4 Ma

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Science china Earth Sciences 2005年第12期48卷 2166-2173页

作者： HUANG Baoqi1,2. JIAN Zhimin2.& WANG Pinxian2.1. school of Earth and Space Sciences, Peking University, Beijing 100871, china 2. Laboratory of Marine Geology, Tongji University, {2. 2.0092. china school of Earth and Space Sciences Peking University Beijing China Laboratory of Marine Geology Tongji University Shanghai China

Upper water structure over the last 4 million years in the northern South china Sea (SCS) was reconstructed by sea surface temperature (SST) and primary productivity estimated by changes in the abundance of planktonic foraminifer, planktonic foraminifer transfer function, and the accumulation rate of benthic foraminifera. Results of SST and primary productivity show that SST gradually decreased since 4 Ma, but differential seasonal temperature and primary productivity increased in steps mainly at about 3.1, 2.7, 2.1, 1.6, 0.9 and 0.5 Ma. Comparison with tectonics and orbital variations indicates that phased rising of the Tibet since 3.6 Ma, closing of Panama and Indonesian seaways, and orbital changes were responsible for the changes of upper water structure in the SCS, respectively.

关键词： northern South china Sea, upper water structure, Late Pliocene, planktonic foraminifer, ODP 1146.

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