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Effect of leukemia inhibitory factor on embryonic stem cell differentiation:implications for supporting neuronal differentiation

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Acta Pharmacologica Sinica 2006年第1期27卷 80-90页

作者： Zhao HE~(2,3) Jing-jing LI~(2,3) Chang-hong ZHEN~(3,4) Lin-ying FENG~3 Xiao-yan DING~(2,5)2 Laboratory of Molecular and Cell Biology and Laboratory of Stem Cell Biology,institute of Biochemistry and Cell Biology:shanghai institutes for Biological sciences,chinese academy of sciences.shanghai 200031,china 3 shanghai institute of materia medica.chinese academy of sciences.shanghai 201203,china 4 Graduate School of the chinese academy of sciences.shanghai 200031,china. LaboratoryofMolecularandCellBiologyandLaboratoryofStemCellBiology InstituteofBiochemistryandCellBiology shanghaiinstitutesforBiologicalsciences ChineseAcademyofSciencesShanghai200031China

Aim:Leukemia inhibitory factor (LIF),a pleiotropic cytokine,has been used ex-tensively in the maintenance of mouse embryonic stem cell *** thiscurrent work,we examined the effect of the LIF signaling pathway in embryonicstem (ES) cell differentiation to a neural ***:In the presence of LIF(1000U/mL),the production of neuronal cells derived from embryoid bodies (EB)was tested under various culture *** of the LIF pathway wasexamined with specific *** effects of cell apoptosis and proliferationon neural differentiation were *** cell differentiation into three-germlayers was ***:Under various culture conditions,neuronal differ-entiation was increased in the presence of *** the LIF-activated STAT3signaling pathway with specific inhibitors abolished the neuronal differentiationof ES cells,whereas inhibition of the LIF-activated MEK signaling pathway im-paired the differentiation of ES cells toward a glial *** suppressed cellapoptosis and promoted cell proliferation during ES cell *** inhib-ited the differentiation of ES cells to both mesoderm and extraembryonic endo-derm fates,but enhanced the determination of neural ***:These results suggest that LIF plays a positive role during the differentiation ofES cells into neuronal cells.

关键词： leukemia inhibitory factor embryonic stem cells neural fate embryoid bodies

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Overcoming multidrug resistance(MDR) in cancer by nanotechnology

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Science china Chemistry 2010年第11期53卷 2226-2232页

作者： BU HuiHui,GAO Yu & LI YaPing shanghai institute of materia medica.chinese academy of sciences.shanghai 201203,china shanghai institute of materia medica Chinese Academy of Sciences Shanghai China

The emerging nanotechnology-based drug delivery holds tremendous potential to deliver chemotherapeutic drugs for treatment of multidrug resistance(MDR) *** drug delivery system could improve the pharmacokinetic behavior of antitumor drugs,deliver chemotherapeutic drugs to target sites,control release of drugs,and reduce the systemic toxicity of drugs in MDR *** review addresses the use of nanotechnology to overcome MDR classified on the bases of the fundamental mechanisms of MDR and various approaches to deliver drugs for treatment of MDR cancer.

关键词： nanotechnology nanoparticles multidrug resistance chemotherapeutic drugs cancer drug delivery system

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Serine racemase expression in mouse cerebral cortex after permanent focal cerebral ischemia

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Acta Pharmacologica Sinica 2004年第4期25卷 38-43页

作者： Li-zhenWANG, Xing-zu ZHU2 Department of Pharmacology, shanghai institute of materia medica, shanghai institutes for Biological sciences, chinese academy of sciences, shanghai 201203, china Department of Pharmacology Shanghai Institute of Materia Medica Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 上海 201203

AIM: To study the alterations of the expressions of serine racemase in C57BL/6 mouse brain after permanent focal cerebral ischemia. METHODS: The mRNA level and the protein level of serine racemase were assayed by semiquantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot, respectively. The amount of D-serine and L-serine were measured by HPLC. RESULTS: High levels of serine racemase were constitutively expressed in the normal cortex of mouse. At early stage after middle cerebral artery occlusion (MCAO), no significant change in expression of serine racemase was observed in temporoparietal cortex in ipsila- teral hemisphere. However, delayed transient decreases of serine racemase in both mRNA and protein levels were detected from d 6 to d 10 after ischemia. Correspondingly, D-serine concentration also declined in the ipsilateral cortex during this period when compared with the D-serine level in the contralateral cortex. CONCLUSION: Delayed decreases in serine racemase expression and D-serine level occurred in the temporoparietal cortex at the late stage after focal cerebral ischemia.

关键词： D-serine L-serine serine racemase inbred C57BL mice brain ischemia

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Progress in studies of huperzine A,a natural cholinesterase inhibitor from chinese herbal medicine

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Acta Pharmacologica Sinica 2006年第1期27卷 1-26页

作者： Rui WANG Hart YAN Xi-can TANG State Key Lahoratory of Drug Research.shanghai institute of materia medica.shanghai institutes for Biological sciences chinese academy of sciences.shanghai 201203,china StateKeyLaboratoryofDrugResearch ShanghaiInstituteofMateriaMedicaShanghaiInstitutesforBiologicalSciencesChineseAcademyofSciencesShanghai201203China

Huperzine A (HupA),a novel alkaloid isolated flom the chinese herb Huperziaserrata,is a potent,highly specific and reversible inhibitor of acetylcholinesterase(ACHE).Compared with tacrine,donepezil,and rivastigmine,HupA has betterpenetration through the blood-brain barrier,higher oral bioavailability,and longerduration of AChE inhibitory *** has been found to improve cognitivedeficits in a broad range of animal *** possesses the ability to protectcells against hydrogen peroxide,β-amyloid protein (or peptide),glutamate,ischemia and staurosporine-induced cytotoxicity and *** protec-tive effects are related to its ability to attenuate oxidative stress,regulate theexpression of apoptotic proteins ***,P53,and caspase-3,protectmitochondria,upregulate nerve growth factor and its receptors,and interfere withamyloid precursor protein *** effects of HupA on N-me-thyl-D-aspartate receptors and potassium currents may also contribute to itsneuroprotection as *** studies in rodents,canines,and healthyhuman volunteers indicated that HupA was absorbed rapidly,distributed widelyin the body,and eliminated at a moderate rate with the property of slow andprolonged release after oral *** and clinical safety tests showedthat HupA had no unexpected toxicity,particularly the dose-limiting hepalotoxic-ity induced by *** phase Ⅳ clinical trials in china have demonstratedthat HupA significantly improved memory deficits in elderly people with benignsenescent forgetfulness,and patients with Alzheimer disease and vasculardementia,with minimal peripheral cholinergic side effects and no *** can also be used as a protective agent against organophosphateintoxication.

关键词： huperzine A Alzheimer disease acetylcholinesterase cholinesterase inhibitors cognitive enhancer neuroprotect agents oxidative stress apoptosis

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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 School of chinese academyof sciences.shanghai 201203,china TheNationalCenterforDrugScreening ShanghaiInstituteofMateriaMedicaChineseAcademyofSciences GraduateSchoolofchineseacademyofsciences shanghai201203 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

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Development of a complex scintillation proximity assay for highthroughput screening of PPAR7 modulators

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Acta Pharmacologica Sinica 2005年第3期26卷 339-344页

作者： Bin WU Jie GAO Ming-wei WANG~2 The National Center for Drug Screening,shanghai institute of materia medica,shanghai institutes for Biological sciences,chinese academy of sciences Graduate School of chinese academy of sciences.shanghai 201203,china The National Center for Drug Screening Shanghai Institute of Materia Medica Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of chinese academy of sciences Shanghai 201203 China

Aim:To develop a complex high-throughput screening （HTS） assay based onscintillation proximity assay （SPA） technology for identification of novel peroxi-some proliferator-activated receptor gamma （PPARγ） ***:Full-length PPARγ and retinoid X receptor alpha （RXRα）,biotinylated PPAR responseelement （PPRE）,[3H]BRL49653 and streptavidin-coated FlashPlate or microbeadwere used to develop an HTS assay based on SPA ***‘ABCDE’method was validated against conventional hydroxyapatite （HA） assay and ap-plied to large-scale screening of 16 000 synthetic compounds and natural ***:（1） IC50values of positive control compounds （BRL49653 andtroglitazone） obtained from the‘ABCDE’method and HA assay were compa-rable and consistent with those reported elsewhere;（2） Approximately 178compounds,showing more than 70% competitive inhibition on BRL49653 bind-ing to PPARγ,were identified initially by the‘ABCDE’method （microbead）;（3）Secondary screening using FlashPlate and cross-reactivity studies with RARα,β,γ and RXRα,β,γ confirmed that 12 compounds possessed specific PPARγ bind-ing properties including 2 with IC50values less than 0.5 μmol/L and novel chemi-cal ***:The‘ABCDE’method using either FlashPlate ormicrobead,is a highly efficient,automatable,and robust tool to screen potentialPPARγ modulators in HTS *** application may be expanded to othernuclear receptors that form heterodimers upon activation.

关键词： peroxisome proliferator-activated receptor gamma retinoid X receptor alpha scintillation proximity assay high-throughput screening

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Expression and purification of lipoprotein-associated phospholipase A2,a key enzyme involved in atherosclerosis

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Acta Pharmacologica Sinica 2006年第6期27卷 679-684页

作者： Fu-jun ZHANG~(2,4)Mao-jun CAI~3 Jing-kang SHEN~3 Yi-ping WANG~(2,5)2 State Key Laboratory of Drug Research.3 Department of Medicinal Chemistry,shanghai institute of materia medica,shanghai institutes for Biological sciences,chinese academy of sciences.shanghai 201203,china 4 Graduate School of the chinese academy of sciences.chinese academy of sciences.shanghai 201203.china StateKeyLaboratoryofDrugResearch DepartmentofMedicinalChemistry ShanghaiInstituteofMateriaMedicaShanghaiInstitutesforBiologicalSciencesChineseAcademyofSciencesShanghai201203China GraduateSchoolofthechineseAc

Aim:To express and purify lipoprotein-associated phospholipase A2（Lp-PLA2）,and to establish a screening model for Lp-PLA2inhibitors using the expressed Lp-PLA2,Methods:We cloned the full-length cDNA of Lp-PLA2 from differentiatedTHP-I cells,and subcloned the cDNA into the baculovirus transfer *** addition,we introduced an N-terminal Kozak sequence for high-level translation initiation and a C-terminal sequence of 6 histidine residues forpurification,The fusion enzyme was expressed in Sf9 insect cells and purified byNi2+affinity *** Lp-PLA2activity was measured us-ing[3H]PAF as a substrate,and we examined the enzyme activity of recombinantLp-PLA2pretreated with the known Lp-PLA2inhibitor ***:Wesuccessfully cloned the full-length Lp-PLA2gene from differentiated THP-1 *** fusion enzyme was expressed in Sf9 insect cells at a high level and purifiedefficiently through a 2-step *** recombinant Lp-PLA2activity wasmeasured using[3H]PAF as a substrate,and proved to be enzymatically ***-PLA2inhibitor SB435495 produced a good inhibition curve for inhibition ofrecombinant Lp-PLA2with an IC50of 57±1μmol/***:We expressed andpurified Lp-PLA2at a high level in insect cell-baculovirus expression *** ratio was much greater than that obtained from human plasma and we estab-lished a screening model for Lp-PLA2inhibitors using the expressed Lp-PLA2.

关键词： lipoprotein-associated phospholipase A2 atherosclerosis coronary heart diseases cloning baculovirus purification

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Down regulation of cyclooxygenase-2 is involved in delayed neuroprotec-tion by ischemic preconditioning in rats

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Acta Pharmacologica Sinica 2005年第4期26卷 441-446页

作者： Liang XIAO Fei-li ZHAO Xing-zu ZHU~1 Department of Pharmacology II,shanghai institute of materia medica,shanghai institutes for Biological sciences,chinese academy of sciences.shanghai 201203,china DepartmentofPharmacologyII ShanghaiInstituteofMateriaMedicaShanghaiInstitutesforBiologicalSciencesChineseA

Aim:To examine whether the prostaglandins(PGs)pathway is involved in trigger-ing delayed neuroprotection by ischemic preconditioning(IPC)and evaluate theeffects of IPC on cyclooxygenase-2(COX-2)expression following focal cerebralischemia and reperfusion in ***:IPC was induced by 10 min of salineinfusion into the left internal carotid artery with the right common carotid arteryclamped at the same *** cerebral artery occlusion(MCAO)and reperfusionmodel was produced using intraluminal filament ***:IPC 48 h priorto MCAO significantly reduced infarct area as compared with MCAO *** inhibitor of COX indomethacin(3 mg/kg ip)applied 1 h prior to or 1 hafter IPC failed to affect its protective *** had no direct effect on thecortex COX-2 mRNA and protein expression 72 h later,but decreased the expres-sion of COX-2 mRNA and protein following ischemia and reperfusion ***:PGs pathways was not involved in triggering delayed neuroprotectionby IPC,and IPC induced down-regulation of COX-2 following focal cerebral is-chemia and reperfusion in rats in vivo.

关键词： ischemic preconditioning brain ischemia prostaglandins cyclooxygenase-2

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Salvicine,a novel topoisomerase Ⅱ inhibitor,exerts its potent anticancer activity by ROS generation

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Acta Pharmacologica Sinica 2007年第9期28卷 1460-1465页

作者： Ling-hua MENG Jian DING Division of Antitumor Pharmacology,State Key laboratory of Drug Research,shanghai institute of materia medica.chinese academy of sciences.shanghai 201203,china Division of Antitumor Pharmacology State Key laboratory of Drug Research Shanghai Institute of Materia Medica Chinese Academy of Sciences Shanghai China

Salvicine is a novel diterpenoid quinone compound obtained by structural modi-fication of a natural product lead isolated from a chinese herb with potent growthinhibitory activity against a wide spectrum of human tumor cells in vitro and inmice bearing human tumor xenografts. Salvicine has also been found to have aprofound cytotoxic effect on multidrug-resisitant (MDR) cells. Moreover, Salvicinesignificantly reduced the lung metastatic foci of MDA-MB-435 orthotopicxenograft. Recent studies demonstrated that salvicine is a novel non-intercalativetopoisomerase II (Topo II) poison by binding to the ATPase domain, promotingDNA-Topo II binding and inhibiting Topo II-mediated DNA relegation and ATPhydrolysis. Further studies have indicated that salcivine-elicited ROS plays acentral role in salvicine-induced cellular response including Topo II inhibition,DNA damage, circumventing MDR and tumor cell adhesion inhibition.

关键词： Salvicine is a novel diterpenoid quinone compound obtained by structural modi-fication of a natural product lead isolated from a chinese herb with potent growthinhibitory activity against a wide spectrum of human tumor cells in vitro and inmice

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Magnesium lithospermate B ameliorates renal cortical microperfusion in rats

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Acta Pharmacologica Sinica 2006年第2期27卷 217-222页

作者： Chun-guang CHEN~(2,3) Yi-ping WANG~(2,4) ~2State Key Laboratory of Drug Research.shanghai institute of materia medica,shanghai institutes for Biological sciences Graduate School of the chinese academy of sciences,chinese academy of sciences.shanghai 201203,china StateKeyLaboratoryofDrugResearch ShanghaiInstituteofMateriaMedicaShanghaiInstitutesforBiologicalSciences GraduateSchoolofthechineseacademyofsciences ChineseAcademyofSciencesShanghai201203China

Aim:To investigate the effects of magnesium lithospermate B（MLB）isolatedfrom Salviae miltiorrhizae on renal microcirculation,and renal and systemic he-modynamics in Sprague-Dawley ***:MLB（10,30,and 60 mg/kg）wasinjected intravenously and renal blood flow（RBF）,renal cortical microperfusion（RCM）,and systemic hemodynamic function parameters including heart rate（HR）,mean arterial pressure（MAP）,left ventricular systolic pressure（LVSP）,left ven-tricular end-diastolic pressure（LVEDP）,and maximal velocity of pressure increase(dp/dtmax)were measured for 45 min after ***:IntravenousMLB at doses of 10,30,and 60 mg/kg increased RCM significantly,but had noobvious effects on RBF or systemic *** effect of MLB on RCMreached its peak 15 rain after injection and returned to baseline after 45 *** to60 mg/kg MLB increased RCM by 62.4%±20.2%（changes from baseline,P3.7%±9.7% vs baseline）and renal vascular resistance（-1.4%±9.1%vs baseline）did not obviously ***:These results indicate thatMLB ameliorates renal microcirculation in a dose-dependent manner,which maybe related to the renoprotective effects of MLB.

关键词： magnesium lithospermate B hemodynamics renal blood flow renal cortical microperfusion

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