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Role of arachidonic acid in hyposmotic membrane stretch-induced increase in calcium-activated potassium currents in gastric myocytes

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Acta Pharmacologica Sinica 2005年第10期26卷 1233-1242页

作者： Meng YANG~2, Xing-lan LI~3, Hui-ying XU~3, Jia-bin SUN~3, Bin MEI~4, Hai-feng ZHENG~3, Lian-hua PIAO~3, De-gang XING~3 Zhai-liu LI~3, Wen-xie XU~(2,5) ~2 Department of Physiology, Shanghai Jiaotong university school of medicine, Shanghai 200030 ~3 Department of Physiology Yanbian university College of medicine Yanji 133000 ~4 East china Normal university school of Life Science, Shanghai 200062, china Department of Physiology Shanghai Jiaotong University School of Medicine Shanghai China Department of Physiology Yanbian University College of Medicine Yanji China East china Normal university school of Life Science Shanghai China

Aim: To study effects of arachidonic acid （AA） and its metabolites on the hyposmotic membrane stretch-induced increase in calcium-activated potassium currents (IKCa) in gastric myocytes. Methods: Membrane currents were recorded by using a conventional whole cell patch-clamp technique in gastric myocytes isolated with collagenase. Results: Hyposmotic membrane stretch and AA increased both IK（Ca）and spontaneous transient outward currents significantly. Exogenous AA could potentiate the hyposmotic membrane stretch-induced increase in IK（Ca）The hyposmotic membrane stretch-induced increase inK（Ca）was significantly suppressed by dimethyleicosadienoic acid （100 μmol/L in pipette solution）, an inhibitor of phospholipase A2. Nordihydroguaiaretic acid, a lipoxygenase inhibitor, significantly suppressed AA and hyposmotic membrane stretch-induced increases in IK（Ca）. External calcium-free or gadolinium chloride, a blocker of stretch-activated channels, blocked the AA-induced increase in IK（Ca）significantly, but it was not blocked by nicardipine, an L-type calcium channel blocker. Ryanodine, a calcium-induced calcium release agonist, completely blocked the AA-induced increase inK（Ca）; however, heparin, a potent inhibitor of inositol triphosphate receptor, did not block the AA-induced increase in IK（Ca）. Conclusion: Hyposmotic membrane stretch may activate phospholipase A2, which hydrolyzes membrane phospholipids to ultimately produce AA; AA as a second messenger mediates Ca2+influx, which triggers Ca2+-induced Ca2+release and elicits activation of IK（Ca）in gastric antral circular myocytes of the guinea pig.

关键词： gastric myocyte arachidonic acid hyposmotic membrane stretch calcium-activated potassium current.

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Phorbol-induced surface expression of NR2A subunit homologues in HEK293 cells

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Acta Pharmacologica Sinica 2006年第12期27卷 1580-1585页

作者： Chan-ying ZHENG~2 Xiu-juan YANG~3 Zhan-yan fu~4 Jian-hong LUO~(2,5)2 Department of Neurobiology,Zhejiang university school of medicine,Hangzhou 310031,china 3 Institute of Neuroscience,Shanghai Institutes for Biological Sciences,the Chinese Academy of Sciences,Shanghai 200031,china 4 Department of Physiology and Biophysics,Georgetown university school of medicine Washington 20057,USA Department of Neurobiology Zhejiang University School of Medicine Hangzhou China Institute of Neuroscience Shanghai Institutes for Biological Sciences the Chinese Academy of Sciences Shanghai China Department of Physiology and Biophysics Georgetown University School of Medicine Washington USA

Aim:N-methyl-D-aspartate receptors(NMDAR)are heteromeric complexes pri-marily assembled from NR1 and NR2 *** normal conditions,NR2 sub-units assemble into homodimers in the endoplasmic reticulum(ER).Thesehomodimers remain in the ER until they coassemble with NR1 dimers and aretrafficked to the cell ***,it still remains unclear whether functionalhomomeric NMDAR exist in physiological or pathological ***:We transfected GFP-NR2A alone into HEK293 cells,treated the cells with PKCactivator 12-myristate-13 acetate(PMA),and then detected surface NR2A sub-units with a live cell immunostaining *** also used a series of NR2Amutants with a partial deletion of its C-terminus to identify the regions that areinvolved in the PMA-mediated surface expression of NR2A ***:NR2A subunits were expressed on the cell membrane after incubation with PMA(200 nmol/L,30 min),although no functional NMDA channels were detected afterPMA-induced membrane *** with an ER marker alsorevealed that NR2A subunits were exported from the ER after PMA ***,the deletion of amino acids between 1149-1347 or 1354-1464 ofNR2A inhibited PMA-induced surface expression ofNR2A ***:First,our data suggests that PMA treatment can induce the surface expression ofhomomeric NR2A ***,this process is probably mediated bythe NR2A C-terminal region between positions 1149 and 1464.

关键词： NMDA receptor NR2A subunit PMA surface expression HEK293 cells

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Antitumor effect of cytosine deaminase/5-fluorocytosine suicide gene therapy system mediated by Bifidobacterium infantis on melanoma

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Acta Pharmacologica Sinica 2005年第5期26卷 629-634页

作者： Cheng YI~2,Ying HUANG~3,Zhi-ying GUO~3,Shu-ren WANG~(3,4) ~2Cancer Center,West china Hospital,Sichuan university Chengdu,610041,china ~3Department of Pathophysiology,West china school of Preclinical and Forensic medicine,Sichuan university,Chengdu 610041,china Cancer Center West China Hospital Sichuan University Chengdu China Department of Pathophysiology West China School of Preclinical and Forensic Medicine Sichuan University Chengdu China

Aim:To construct a Bifidobacterium infantis/CD targeting gene therapy systemand observe the antitumor effect of cytosine deaminase/5-fluorocytosine(CD/5-FC)suicide gene therapy system mediated by Bifidobacterium infantis on mela-noma in vitro and in ***:A recombinant CD/pGEX-1LamdaT plasmidwas transfected into Bifidobacterium infantis by ***-terium infantis transfected by recombinant CD/pGEX-1LamdaT plasmid was in-cubated with 5-FC *** the supernatant fluid was collected andadded to melanoma B 16-F10 cells to observe the killing effect for B 16-F10 *** were inoculated with melanoma B 16-F10 cells to establish animal *** mice were then injected with 5-FC and Bifidobacterium infantis transfectedby recombinant CD/pGEX-1LamdaT ***:Two segments of approxi-mate 4.9 kb and 1.3 kb were extracted from the 6.2 kb recombinant plasmid,whichwere equal to the size of the pGEX-1LamdaT plasmid and CD gene,*** results showed that the full length and sequence of nucleotide acid ofthe inserted gene in extracted recombinant plasmid was completely identical to theCD *** vitro,B16-F10 cells treated by supernatant fluid were remarkablydamaged morphologically,and the cell growth was significantly ***-ments on the mice melanoma model showed that after treatment with a combina-tion of transfected Bifidobacterium infantis and 5-FC,the tumor volume wassignificantly inhibited compared with ***:The foreign gene,CD gene,was correctly inserted into pGEX-1LambdaT plasmid and transferredinto Bifidobacterium ***/5-FC suicide gene therapy system mediated byBifidobacterium infantis demonstrated a good antitumor effect on melanoma invitro and in vivo.

关键词： Bifidobacterium infantis cytosine deaminase gene therapy

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Animal models of Huntington's disease: implications in uncovering pathogenic mechanisms and developing therapies

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Acta Pharmacologica Sinica 2006年第10期27卷 1287-1302页

作者： Lin-hui WANG~2 Zheng-hong QIN~(3,4) Departments of ~2Physiology and ~3Pharmacology,Soochow university school of medicine,Suzhou 215123,china Department of Physiology Soochow University School of Medicine Suzhou China Department of Pharmacology Soochow University School of Medicine Suzhou China

Huntington’s disease (HD) is an autosomal dominant neurodegenerative disorder,which is caused by an abnormal expansion of Cytosine Adenine Guanine (CAG)trinucleotide repeat in the gene making huntingtin (Htt).Despite intensiveresearch efforts devoted to investigate molecular mechanisms of pathogenesis,effective therapy for this devastating disease is still not available at *** of various animal models of HD has offered alternative approachesin the study of HD molecular *** HD models,including chemical-induced models and genetic models,mimic some aspects of HD symptoms *** date,however,there is no ideal model which replicates all of theessential features of neuropathology and progressive motor and cognitive impair-ments of human *** a result,our understanding of molecular mechanisms ofpathogenesis in HD is still limited.A new model is needed in order to uncover thepathogenesis and to develop novel therapies for *** this review we discussedusefulness and limitations of various animal and cellular models of HD in uncov-ering molecular mechanisms of pathogenesis and developing novel therapies forHD.

关键词： Huntington’s disease huntingtin neurodegeneration animal models

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Magnetic resonance imaging based volumetry:a primary approach to unravelling the brain

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Progress in Natural Science:Materials International 2007年第7期17卷 751-760页

作者： Huang Xiaoqi~(1,2) LüSu~1 Li Dongming~1 Gong Qiyong~(1,3**) (1.Huaxi MR Research Centre(HMRRC),and Department of Radiology,West china Hospital/West china school of medicine,Chengdu 610041,china 2.Department of Psychiatry,West china Hospital/West china school of medicine,Chengdu 610041,china 3.Division of Medical Imaging,Faculty of medicine,university of Liverpool,United Kingdom) Huaxi MR Research Centre (HMRRC) Department of Radiology West China Hospital/West China School of Medicine Chengdu 610041 China Department of Psychiatry West China Hospital/West China School of Medicine Chengdu 610041 China Division of Medical Imaging Faculty of medicine University of Liverpool United Kingdom United Kingdom

Magnetic resonance(MR)imaging based volumetry is recognized as an important technique for studying the *** this review,two principle volumetric methods using high resolution MR images were introduced,namely the Cavalieri method and the voxel based morphometry(VBM).The Cavalieri method represents a manual technique that allows the volume of brain structures to be estimated efficiently with no systematic error or sampling bias,whereby the VBM represents an automated image analysis which involves the use of statistical parametric mapping of the MR imaging *** methods have been refined and applied extensively in recent neuro- science *** present paper aims to describe the development of methodologies and also to update the knowledge of their applica- tions in studying the normal and diseased brain.

关键词： MR imaging Cavalieri method voxel-based morphometry brain volumetry

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Altered subcellular distribution of nucleolar protein fibrillarin by actinomycin D in HEp-2 cells

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Acta Pharmacologica Sinica 2004年第7期25卷 56-60页

作者： Min CHEN2, Ping JIANG Research Center of Basic medicine, school of medicine, Jiangsu university, Zhenjiang 212001, china Research Center of Basic medicine School of Medicine Jiangsu University Zhenjiang 212001

AIM: To study the effects of actinomycin D on subcellular distribution of nucleolar protein fibrillarin in HEp-2 (human esophageal epithelial type 2) cells, and molecular mechanisms for maintenance of fibrillarin in nucleolus. METHODS: Indirect immunofluorescence assay was employed to investigate subcellular distribution of nucleolar protein fibrillarin and immunoblotting analysis was used to detect the total cellular amount of fibrillarin. RESULTS: Control cells with no drug treatment showed bright clumpy nucleolar staining, which indicated that fibrillarin decorated the nucleolus only. Treatment with actinomycin D caused dislocation of fibrillarin from nucleoli to nucleoplasm with numerous stained small nucleoplasmic entities. Immunoblotting showed that neither total cellular amount of fibrillarin nor the integrity of fibrillarin was changed upon the treatment. The dislocation of fibrillarin in cells treated at a lower concentration (0.05 mg/L) of actinomycin D, was totally reversible after removal of the drug from the medium. However, this reversion was not observed at a high drug concentration (1 mg/L). CONCLUSION: Actinomycin D induced dislocation of fibrillarin from nucleoli to nucleoplasm in HEp-2 cells. The retention of fibrillarin within the nucleolus was related to active RNA synthesis.

关键词： dactinomycin nuclear proteins fluorescence antibody technique immunoblotting

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Effects of GSM 1800 MHz on dendritic development of cultured hippocampal neurons

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Acta Pharmacologica Sinica 2007年第12期28卷 1873-1880页

作者： Wei NING~2,Shu-jun XU~2,Huai CHIANG~3,Zheng-ping XU~3,Su-ya ZHOU~2,Wei YANG,Jian-hong LUO~(2,4)~2 Department of Neurobiology,Zhejiang university school of medicine,Hangzhou 310058,china ~3 Bioelectromagnetics Laboratory,Zhejianguniversity school of medicine,Hangzhou 310058,china Department of Neurobiology Zhejiang University School of Medicine Hangzhou China Bioelectromagnetics Laboratory Zhejiang University School of Medicine Hangzhou China

Aim:To evaluate the effects of global system for mobile communications (GSM)1800 MHz microwaves on dendritic filopodia,dendritic arborization,and spinematuration during development in cultured hippocampal neurons in ***:The cultured hippocampal neurons were exposed to GSM 1800 MHz microwaveswith 2.4 and 0.8 W/kg,respectively,for 15 min each day from 6 days in vitro (DIV6)to *** subtle structures of dendrites were displayed by transfection withfarnesylated enhanced green fluorescent protein (F-GFP) and GFP-actin on DIV5into the hippocampal ***:There was a significant decrease in thedensity and mobility of dendritic filopodia at DIV8 and in the density of maturespines at DIV14 in the neurons exposed to GSM 1800 MHz microwaves with2.4 W/*** addition,the average length of dendrites per neuron at DIV10 andDIV14 was decreased,while the dendritic arborization was unaltered in ***,there were no significant changes found in the neurons ex-posed to the GSM 1800 MHz microwaves with 0.8 W/***:Thesedata indicate that the chronic exposure to 2.4 W/kg GSM 1800 MHz micro-waves during the early developmental stage may affect dendritic developmentand the formation of excitatory synapses of hippocampal neurons in culture.

关键词： global system for mobile communications microwaves dendritic filopodia dendritic arborization spines hippocampal neurons

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Suppression of Type 1 Insulin-like Growth Factor Receptor Expression by Small Interfering RNA Inhibits A549 Human Lung Cancer Cell Invasion in vitro and Metastasis in Xenograft Nude Mice

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Acta Biochimica et Biophysica Sinica 2007年第2期39卷 137-147页

作者： Jianfang QIAN~1 Aiqiang DONG~1 Minjian KONG~1 Zhiyuan MA~2 Junqiang FAN~1 and Guanyu JIANG~3 ~1Department of Cardiothoracic Surgery,Second Affiliated Hospital,school of medicine,Zhejiang university,Hangzhou 310009,china ~2Department of Cardiovascular Surgery,Shanghai Jiaotong university Affiliated First People's Hospital,Shanghai 200080,china ~3Department of Emergency,Second Affiliated Hospital school of medicine,Zhejiang university,Hangzhou 310009,china Department of Cardiothoracic Surgery Second Affiliated Hospital School of Medicine Zhejiang UniversityHangzhou 310009 China Department of Cardiovascular Surgery Shanghai Jiaotong University Affiliated First People's HospitalShanghai 200080 China Department of Emergency Second Affiliated Hospital School of Medicine Zhejiang UniversityHangzhou 310009 China

Cancer invasion and metastasis, involving a variety of pathological processes andcytophysiological changes,contribute to the high mortality of lung *** type 1 insulin-like growthfactor receptor (IGF-1R),associated with cancer progression and invasion,is a potential anti-invasion andanti-metastasis target in lung *** inhibit the invasive properties of lung cancer cells,we successfullydown-regulated IGF-1R gene expression in A549 human lung cancer cells by small interfering RNA (siRNA)technology,and evaluated its effects on invasion-related gene expression,tumor cell in vitro invasion,andmetastasis in xenograft nude mice.A549 cells transfected with a plasmid expressing hairpin siRNA forIGF-1R showed a significantly decreased IGF-1R expression at the mRNA level as well as the *** biological assays,transfected A549 cells showed a significant reduction of cell-matrix adhesion,migration and *** with these results,we found that down-regulation of IGR-1Rconcomitantly accompanied by a large reduction in invasion-related gene expressions,including MMP-2,MMP-9,u-PA,and IGF-1R specific downstream *** tail vein injections of plasmid expressinghairpin siRNA for IGF- 1R significantly inhibited the formation of lung metastases in nude *** resultsshowed the therapeutic potential of siRNA as a method for gene therapy in inhibiting lung cancer invasionand metastasis.

关键词： lung cancer invasion metastasis type 1 insulin-like growth factor receptor small interfering RNA

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Inactivation of TGF-βsignaling in CAR-T cells

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Cellular & Molecular Immunology 2024年第3期21卷 309-310页

作者： Zaopeng Yang Yang-Xin fu Changping Laboratory Beijing102206China Department of Basic Medical Sciences School of MedicineTsinghua UniversityBeijing100084China

Accumulating evidence suggests that the transforming growth factor(TGF)-βpathway might be an ideal target for optimizing chimeric antigen receptor(CAR)-T-cell therapy for solid *** Cellular&Molecular Immunology,Liang et al.(2024)reported that CAR-T cells expressing SMAD7 displayed favorable antitumor efficacy and a reduced risk of systemic toxicity.

关键词： CAR Immunology

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Antioxidant Effects of Dehydroepiandrosterone Are Related to Up-regulation of Thioredoxin in SH-SY5Y Cells

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Acta Biochimica et Biophysica Sinica 2005年第2期37卷 119-125页

作者： Jing GAO Hui-Ying SUN Zeng-Rong ZHU Zhen DING Li ZHU school of medicine.school of Life Sciences,Nanjing university.Nanjing 210093.china Institute of Nautical medicine,Nantong university.Nantong 226001,china school of medicine School of Life Sciences Nanjing UniversityNanjing 210093 China Institute of Nautical medicine Nantong UniversityNantong 226001 China

Neuroprotective effects of dehydroepiandrosterone（DHEA）have been shown to be associatedwith its antioxidant properties,but the mechanisms remain *** that the thioredoxin（Trx）system,an important cellular redox modulation system,changes under oxidative stress and could exertprotective effects,the relationship between the antioxidant effects of DHEA and the Trx system regulationwas *** MTT assay and morphological observation,the effects of DHEA in the model of H2O2-induced oxidative stress in SH-SY5Y cells were analyzed,then RT-PCR and Western blot assay were used todetect the alteration in mRNA and protein level of Trx,The results showed that a pre-treatment of DHEA（10-100nM）protected cells against the toxicity induced by H2O2in a dose-dependent manner,which could beconfirmed in morphological observation by phase-contract *** addition,Trx mRNA transcriptionwas inhibited by H2O2（300μM）,which could be reversed by the pre-administration of DHEA in variousconcentrations（0.1-100nM）.Western blot assay confirmed that protein level of Trx could be elevated by thepre-treatment of DHEA（10-100nM）with the exposure of *** together,these data suggest thatDHEA may be useful in treating age-related neurodegenerative diseases based on its up-regulating effects onan antioxidant and neuroprotective protein thioredoxin,a substrate in the Trx redox system.

关键词： dehydroepiandrosterone(DHEA) thioredoxin(Trx) H2O2 SH-SY5Y cell

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