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High-Throughput Screening of Flavonoids Inhibitors of Catechol-O-Methyltransferase with a Two-Photon Fluorescent probe

High-Throughput Screening of Flavonoids Inhibitors of Catech...

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中国药理学会分析药理学专业委员会成立大会、第三届全国分析药理学学术大会暨贵州省药学会药学青年专业委员会成立大会

作者： Fangyuan Wang Ping Wang Guangbo Ge Ling Yang Shanghai University of Traditional Chinese Medicine

Aim:Catechol O-methyltransferase（COMT）,which catalyzes the catechol compounds to O-methylated metabolites,plays a key role in the metabolism of endogenous catecholamine neurotransmitters and dopaminergic drug such as *** of human brain COMT is an important therapy approach for Parkinson's disease,because the potent COMT inhibitors can slow down the metabolism of levodopa and dopamine,resulting in the increase of dopamine level in the *** flavonoids have been long recognized as an interesting class of COMT inhibitors due to their catechol skeletons and abilities to bind the active sites of the ***:We first synthesize a two-photon fluorescent probe,3-BTD,for determination of COMT activity in various biological samples and bio-imaging of endogenous COMT in the living cells,which displays good reactivity and high specificity to the ***,a high-throughput screening method is developed for COMT inhibitors from natural flavonoids using the 3-BTD as a small-molecule *** further inhibition kinetics assays demonstrate the inhibition potentials of these flavonoid ***:In this study,the C-8 phenolic hydroxyl of 3-BTD is selectively methylated by COMT,and the corresponding product,3-BTMD,exhibits about 150-fold fluorescence enhancement at 380 nm excitation/510 nm *** fluorescence quantum yields of 3-BTD and 3-BTMD are determined to be 0.0003 and 0.673,*** 3-BTD probe has been successfully applied to the high-throughput screening of COMT inhibitors,and the inhibitory effects of 50 flavonoid compounds are *** them,only 3 compounds（baicalein,scutellarein,and hispidulin） exhibit stronger inhibitory effects on COMT activity,with the IC values of 0.039,0.053 and 0.036 μM,***:3-BTD exhibited extremely weak fluorescent signal because of the interactions between the photo-induced electron transfer of 8-phenolic hydroxyl and the intramolecular charge transfer of 7-phenolic h

关键词： Catechol O-methyltransferase(COMT) high-throughput screening natural flavonoids two-photon fluorescent probe

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Regulation of key genes in bile acid pathways by chenodeoxycholic acid in primary hepatocytes

Regulation of key genes in bile acid pathways by chenodeoxyc...

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中国药理学会分析药理学专业委员会成立大会、第三届全国分析药理学学术大会暨贵州省药学会药学青年专业委员会成立大会

作者： Yanping Yang Lin Qin Yimei Du Yuqi He School of Pharmacy Zunyi Medical University The Key Laboratory of Basic Pharmacology of Ministry of Education Zunyi Medical University

AIM:To investigate the regulation of key genes in the bile acid pathways by farnesoid X receptor(FXR) agonist,chenodeoxycholic acid(CDCA) in primary mouse ***:C57 BL/6 mouse hepatocytes were isolated by modified two-step collagenase perfusion method,purified by density gradient centrifugation,and seeded at the density of 1×10～6 cells/well in a 6-well culture *** treated with CDCA at a final concentration of 25,50 and100 μmol/L for 24 *** genes expression of cholesterol 7-alpha hydroxy-lase(Cyp7 al),sterol12 a-hydroxylase(Cyp8 b1),organic solute transporter beta(OstP) and small heterodimer partner(Shp) were detected by reverse transcription-polymerase chain reaction(RT-PCR).RESULTS:Mouse hepatocytes were successfully isolated and viability was over 80%.Compared with control *** with increasing CDCA *** bile acid synthesis gene,100 μmol/L CDCA significantly upregulated the messenger RNA(mRNA) expression of Cyp7α1 in mouse hepatocytes(P<0.05),25 and 50 umol/L CDCA not significantly differ(P>0.05).25 and 100μmol/L CDCA significantly downregulated the mRNA expression of Cyp8 bl in mouse hepatocytes(P<0.05),however,50 umol/L CDCA seem to have no effect(P>0.05).Regulation of bile acid synthesis genes in the liver pathway,100 umol/L CDCA significantly upregulated the messenger RNA(mRNA) expression of Shp in mouse hepatocytes(P<0.05),25 and 50 umol/L CDCA not significantly differ(P>0.05).50 and 100 umol/L CDCA significantly upregulated the mRNA expression of Ostβ in mouse hepatocytes(P<0.05),and 25 umol/L CDCA not significantly differ(P>0.05).All of this indicates that FXR ligand — CDCA plays an important role in modulating the metabolism of bile acids,and the regulation of CDCA is most obvious at 100 umol/***:CDCA significantly changes expression levels of genes involved in bile acids homeostasis regulation in primary mouse hepatocytes.

关键词： chenodeoxycholic acid primary mouse hepatocytes Bile acid key genes

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Population Pharmacokinetics and Dosing Regimen Optimisation of Lopinavir in Chinese Adults Infected with HIV

Population Pharmacokinetics and Dosing Regimen Optimisation ...

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中国药理学会分析药理学专业委员会成立大会、第三届全国分析药理学学术大会暨贵州省药学会药学青年专业委员会成立大会

作者： NIU Wan-jie SUN Tao LIU li LIU Xiao-qian ZHANG Ren-fang YIN Lin WANG Jiang-rong JIA Xiao-fang LU Hong-Zhou ZHONG Ming-kang JIAO Zheng ZHANG Li-jun Department of Pharmacy Huashan HospitalFudan University

Objectives:Lopinavir（LPV） is a protease inhibitor（PI） for the treatment of human immunodeficiency virus（HIV） *** studies on LPV are mainly focused on Caucasians,and none have investigated the population pharmacokinetics（PPK） of LPV in Chinese *** present study aimed to develop a PPK model for oral LPV in a Chinese adults who are ***:A total of 460 LPV concentrations from 174 Chinese patients who received LPV/ritonavir（LPV/r） 400/100 mg orally every 12 hours（ql2 h） were analysed using the nonlinear mixed effects modelling *** of the LPV concentration profile were performed with different dose regimens.A one-compartment model with first-order absorption and elimination process described the ***:The estimated apparent clearance（CL/F） and volume of distribution（V/F）(%relative standard error[RSE]) for oral LPV were 5.9 L/h（3%） and 117 L（8%）,*** weight was identified as a covariate on CL/*** patients who weighted between 45 and 115 kg and received the standard400/100 mg ql2 h regimen,the probability of achieving target trough concentration(C) of1 mg/L was >98%for PI-naive patients and the probability of achieving target C of 4 mg/L was <80%for PI-pretreated ***:This is the first population pharmacokinetics study to characterise the PK of LPV in Chinese patients with HTV *** were no obvious ethnic differences in the PK of LPV between the Chinese and Caucasian *** simulations demonstrated that the standard dose regimen of 400/100 mg ql2 h（LPV/r tablets）appears to be sufficient for Pi-naive patients but suboptimal for Pl-pretreated ***,the regimen of 800/200 mg ql2 h was recommended for Pl-pretreated *** further investigation of dosages recommendation could be helpful in optimizing LPV therapy for HTV.

关键词： lopinavir population pharmacokinetics HIV Chinese patients simulation

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Effects of Dendrobium nobile *** intestinal transporters based on Metabolomics

Effects of Dendrobium nobile *** intestinal transporters bas...

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中国药理学会分析药理学专业委员会成立大会、第三届全国分析药理学学术大会暨贵州省药学会药学青年专业委员会成立大会

作者： LIU Hao ZHANG Yi-xin LING Lei FENG Ze-yu ZENG Xing-xing HE Yu-qi LU Yuan-fu LU Yan-liu Key Laboratory of Basic Pharmacology of Ministry of Education and Joint International Research Laboratory of Ethnomedicine of Ministry of Education Zunyi Medical University Pharmaceutical Lab National Demonstration Center for Experimental(Pharmacy) EducationSchool of PharmacyZunyi Medical Univesity

Dendrobium nobile *** a famous traditional Chinese herb used for thousands of years in *** characteristics of long term clinical usage make safety hazard could be potential through drug-drug interaction,however,without clear *** present research was designed to investigate the effects of *** on intestinal transporters using metabolomics analysis,as well as provide a prediction for potential drug ***-four rats were randomly divided into normal control group and *** treatment group（0.25 g/kg）.The rats were intragastrically administrated once every 6 h for 5 *** the last administration,the rats were fasted for 6 *** was drawn from the orbit in 6 h after the last treatment,and then centrifuged at 4500 rpm（4 ℃,15 min）.The plasma obtained was stored at -80 °C for later metabolomics *** small intestine tissue was dissected and immediately frozen at-80 ℃for future *** plasma samples were detected with Thermo QE *** chromatography separation was performed on a Hypersil Gold C column（150 × 2.1 mm,1.9 um）with acetonitrile-0.1%formic acid in water by gradient *** ionization（ESI）source was applied and operated in positive ion *** component analysis（PCA） and cluster analysis were used to analyze the metabolomics *** with the normal control group,the different metabolites in *** treatment group were identified,and then made the KEGG pathway *** expression levels of intestinal transporter genes were studied by the quantitative real-time PCR for *** results showed that dendrobine,mubironine B and dendramine,which are the main alkaloid compounds of ***,were detected in plasma samples after oral *** suggested that dendrobine,mubironine B and dendramine may be the main active substances for *** results of PCA and cluster analysis showed that there were obvious differences between the normal control group and ***

关键词： Dendrobium nobile Lindl. Metabolomics Intestinal transporter

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Triptolide inhibits stimulatory effects of radiation lung rigidity on myofibroblasts activation

Triptolide inhibits stimulatory effects of radiation lung ri...

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中国药理学会分析药理学专业委员会成立大会、第三届全国分析药理学学术大会暨贵州省药学会药学青年专业委员会成立大会

作者： Chun Chen Shanmin Yang Kaining Guo Yanwei Zhang Zhangjie Chen Mei Zhang Jianhua Xu Paul Okunieff Lurong Zhang Department of Pharmacology School of PharmacyFujian Medical University School of Pharmacy Fujian Key Laboratory of Natural Medicine PharmacologyFujian Medical University Institute of Materia Medica School of PharmacyFujian Medical University(FMU) Department of Radiation Oncology University of Florida Fujian Key Lab of Individualized Active Immunotherapy and Key Laboratory of Radiation Biology of Fujian Province Universities

OBJECTIVE:Excessive myofibroblast(MFB) activation results in massive extracellular matrix deposition and *** rigidity is the main microenvironmental factor that promotes MFB *** study demonstrated that Triptolide(TPL) reduced MFB activation in radiation-induced lung fibrosis(RILF).This study further explored if TPL inhibits the stimulatory effects of radiation lung rigidity on MFB ***:By using a co-culture system which mimics the in vivo environment,an in vitro 3 D matrix system and a mouse model of RILF,we observed the regulation of TPL on lung tissue matrix stiffness and MFB *** data indicated that the increased levels of Collagen I,LOX and LOXL-2 in RILF were the main causes of tissue rigidity,which was inhibited by *** core mechanism of stiff matrix stimulating MFB activation was mainly through integrins/FAK/RhoA pathway validated by analysis of comparative transcriptome,immunofluorescence and Western blotting,which was also inhibited by ***:These results demonstrate that anti-RILF effects of TPL is associated with multi-target inhibition of stimulatory effects of matrix rigidity on MFB activation.

关键词： radiation-induced lung fibrosis(RILF) matrix stiffness myofibroblast(MFB) triptolide(TPL)

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Dentrobium Alkaloids Promotes Neural Function after Cerebral Ischemia-Reperfusion Injury through Inhibiting Pyroptosis Induced Neuronal Dearth in Vivo and Vitro Models

Dentrobium Alkaloids Promotes Neural Function after Cerebral...

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中国药理学会分析药理学专业委员会成立大会、第三届全国分析药理学学术大会暨贵州省药学会药学青年专业委员会成立大会

作者： Daohang Liu Zhi Dong Chongqing Key Laboratory of Biochemistry and Molecular Pharmacology College of PharmacyChongqing Medical University

Pyroptosis is a new-found lytic form of programmed cell death of cerebral ischemia injury that contributes to the activation and secretion of inflammatory cytokine IL-1β,IL-18 and IL-6,which also characterizes by plasma membrane blebbing prior to *** research have found that Dendrobium Alkaloids can exert neuroprotective effect against OGD/R damage in vitro,but the mechanisms remain *** this study,we detected whether Dendrobium Alkaloids show a therapeutic benefits to I/R damage vie ameliorating pyroptosis and ***22 cells pretreated with Dendrobium Alkaloids(0.03,0.3,3 mg/ml,24 h in advance) were involved in oxygen-glucose deprivation andre-oxygenation(OGD/R).Neuronal apoptosis,neuronal necrosis,neuronal pyrotosis,neuronal pathologic changes and tight junction protein Pannexin-1 were analyzed at 24 h followeing OGD/*** for a further investigation,Male C57/BL mice pretreated with Debdrobium Alkaloids at different concentration(0.5,Smg/ml,ip.) for 24 hours,MCC950(50 mg/kg,ip.) and VX765(50 mg/kg,ip.) undwent transient middle cerebral artery occlusion and ***,we verified Drendrobium Alkaloids pretreatment facilitated lower neurologic deficit score,less infarct volume,fewer pyroptotic cells,reductive inflammatory factors levels and degraded pyroptosis-related proteins at 24 hours after I/*** is highlight that pyroptosis-associated Caspase-1 DSDMD and GSDMD-C protein level were also decreased by Denreobium Alkaloids administration,particularly in the hippocampal CA1 region,and the same decrease was visible in IL-1β,IL-6 and IL-18 that were known for their significant inflammatory participant *** junction protein(Pannexin-l) were down-regulated by Dendrobium Alkaloids,which underlined the improvement of ultrastructural damage associated with OGD/***,the findings demonstrated Dendrobium Alkaloids serve a potentially therapeutic effect against cerebral ischemia damage through reducing pyroptotic neurons,providing ne

关键词： Dendrobium Alkaloids Pyroptosis Inflammation Ischemic-reperfusion injury Caspase-1 GSDMD Membrane pores

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Ginsenoside Rh2 enhances anti-tumor immunity through improving liver fatty acid metabolism in non-small cell lung cancer mice

Ginsenoside Rh2 enhances anti-tumor immunity through improvi...

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中国药理学会分析药理学专业委员会成立大会、第三届全国分析药理学学术大会暨贵州省药学会药学青年专业委员会成立大会

作者： Ming Xiang Ying Qian Rongrong Huang Senlin Li Rui Xie Department of Pharmacology School of PharmacyTongji Medical CollegeHuazhong University of Science and Technology

Aims:Ginsenoside Rh2(G-Rh2) as a natural active component extracted from traditional Chinese medicine ginseng,exhibits rich pharmacological activity,including anti-tumor *** the research we aimed to study the therapeutic effects and involving mechanism of G-Rh2 and its combination with cyclophosphamide(CY) on non-small cell lung cancer(NSCLC) of ***:We established a NSCLC subcutaneous xenograft model of C57 BL/6 j mice and nude mice to measure the anti-tumor effects of G-Rh2,CY and the combination of G-Rh2 and *** observe the tumor growth we recorded tumor volume every day and used in vivo imaging technology to track the accumulation of cancer cells in *** enzyme-linked immunosorbent assay and quantitative real-time polymerase chain reaction(qPCR) we analyzed fatty acid(FA)metabolism in *** applied flow cytometry and mixed lymphocyte reaction to study the immune function of *** we explored the cross-talk between hepatocytes and immune cells by ***:The result showed G-Rh2 had slight anti-cancer effect,while,once combined with CY,the synergistic therapeutic benefit was significantly enhanced with a relief of adverse ***,we found G-Rh2 alleviated abnormal FA metabolism of liver including inhibiting the synthesis of FA and promoting the oxidation of *** improvement of FA metabolism contributed to the enhancement of immune function which promoted the anti-tumor effect of *** cell cross-talk assay also indicated G-Rh2 enhanced function and differentiation of immune cells through affected the metabolism of *** research showed G-Rh2 decreased the expression of forkhead box gene,group O-1(FOXO1) which inhibited the expression of fatty acid synthase(FASN) to decrease FA production and increase oxidative decomposition of FA in ***:Our research found G-Rh2 played anti-NSCLC effect in part by improved FA metabolism to enhance immunity via FoxO1-FASN *** once combined with CY,the

关键词： Ginsenoside Rh2 Synergistic treatment Fatty acid metabolism Immunity

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Oleanolic acid and ursolic acid induce UGT1A1 expression by activating PXR rather than CAR

Oleanolic acid and ursolic acid induce UGT1A1 expression by ...

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中国药理学会分析药理学专业委员会成立大会、第三届全国分析药理学学术大会暨贵州省药学会药学青年专业委员会成立大会

作者： Na Yao Caiwen Zeng Tao Zhan Li Yuan Lingna Wu Hong Zhang Yuqing Xiong Chunhua Xia Clinical Pharmacology Institute Nanchang University Jiangxi Province Cancer Hospital

Background:Oleanolic acid(OA) and its isomer ursolic acid(UA) have recently emerged as research foci based on their biologic *** previously demonstrated that UA can inhibit the activities of UGT1 A3 and UGT1 A4,and OA impedes UGT1 A3 activity in liver ***,whether OA and UA affect the expression of UGT1 As in HepG2 cells and the underlying regulatory mechanism remain ***:The present study aimed to explore the effect of OA and UA on the expression of UGT1 As in HepG2 cells and the regulatory mechanisms on UGT1 A1 based on the pregnane X receptor(PXR) and constitutive androstane receptor(CAR) signaling ***:We analyzed the effect of OA and UA on UGT1 A expression and on the PXR/CAR regulatory pathway in HepG2 cells,in hPXR-silenced or hCAR-silenced HepG2 cells by Q-PCR,Western blotting,and dual-luciferase reporter gene ***:In HepG2 cells,OA and UA both significantly induced the expression of UGT1 A1,UGT1 A3,UGT1 A4,and UGT1 A9 and upregulated the expression of ***,OA and UA did not affect CAR expression.A dual-luciferase reporter assay showed that OA and UA could markedly promote PXR-mediated UGT1 A1 luciferase activity,whereas OA and UA did not affect CAR-mediated UGT1 A1 luciferase *** hPXR-silenced HepG2 cells,OA and UA did not elevate UGT1 A1 activity compared to the control ***,the expression of UGT1 A1 in hCAR-silenced HepG2 cells was markedly elevated compared to the control group or with non-silenced HepG2 cells treated with OA(10,20,and 40 μM) or UA(10,20,and 40 μM).Conclusions:OA and UA significantly induce the expression of UGT1 A1,UGT1 A3,UGT1 A4,and UGT1 A9 in HepG2 cells,and their induction on UGT1 A1 is mediated by PXR activation,not CAR.

关键词： OA UA UGT1A1 PXR CAR

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Fecal samples of Kazaks normal glucose tolerance modulates gut microbiota during alleviation of type 2 diabetes in db/db mice

Fecal samples of Kazaks normal glucose tolerance modulates g...

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中国药理学会分析药理学专业委员会成立大会、第三届全国分析药理学学术大会暨贵州省药学会药学青年专业委员会成立大会

作者： Ye Wang zhangpeipei hanxue luoxin Yicun Tao Xinmin Mao Linlin Li Pharmacological Department Basic Medicine CollegeXinjiang Medical University

Objective:The fecal samples of normal glucose tolerance in Kazak donor（KNGT） can improve the db/db mice symptoms in our previous *** this article we explore the fecal microbiota transplantation（FMT） of KNGT in regulating the intestinal microflora and unravel the mechanisms by which KNGT can maintain intestinal flora homeostasis in db/db ***:db/m and db/db mice were divided into M0,D0,M6,D6,H6 groups.（M0:db/m mice at Oweek.M6:db/m mice with PBS at 6 ***:db/db mice at 0 week.D6:db/db+PBS at 6 weeks.h6:db/db+ KNGT at 6 weeks,n=6/group）.Stool samples were collected 0 and 6 week for extracting genomic DNA by means of a commercial kit from *** V4 variable region of the16 S rRNA gene was subjected to high-throughput sequencing by MiSeq *** Qualified libraries are sequenced pair end on the MiSeq System with the sequencing strategy PE 250 or PE300 MiSeq Reagent *** obtained data were analyzed using bioinformatic tools and multivariable statistical ***:NMDS analysis showed the sample of H6 group was clustered in the middle and separated DO,D6 and m0、 m6 groups,suggesting that the community structure of db/db mice migrated to normal mice under the intervention of *** OTUs were significantly associated with the bacterial community structure in the intestine of the 5 groups of mice as determined by linear discriminant analysis effect size（LEfSe）*** are clostridiaceae-Clostridium,paraprevotella and F16,TM7,CW040,TM7-handsome were enriched in MO ***-Burkholderiales-Alcaligenaceae-Sutterella,s24-7,Ruminococcusflavefaciens were in M6 ***-clostridiales,firmicutesRuminococcaceae,mycoplasmatales-mycoplasmataceae-Mycoplasma were in D0;H6 group just showed selective enrichment in Rikenella community,while D6 showed selective enrichment community including enterobacteriales-enterobacteriaceae-escherichia-escherichiaoli;At each classification level,the different composition ratio of each strain

关键词： Fecal samples Kazaks normal glucose tolerance intestinal flora 16S rDNA db/db mice

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