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Developmental regulation of intracellular calcium transients during cardiomyocyte differentiation of mouse embryonic stem cells

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Acta Pharmacologica Sinica 2006年第7期27卷 901-910页

作者： Ji-dong FU Hui-mei YU Rong WANG Ji LIANG Huang-tian YANG~2.key laboratory of Stem Cell Biology,Institute of Health Sciences,{2. Institutes for Biological Sciences,Chinese Academy of Sciences and {2. Jiao Tong University School of medicine.{2. 2.002.,china keylaboratoryofStemCellBiology InstituteofHealthSciencesShanghaiInstitutesforBiologicalSciencesChineseAcademyofSciencesandShanghaiJiaoTongUniversitySchoolofMedicineShanghai200025China

Aim:To investigate the developmental regulation of intracellular Ca2.transients,an essential event in excitation-contraction coupling,during ***:Using the embryonic stem(ES)cell in vitro differentia-tion system and pharmacological intervention,we investigated the molecular andfunctional regulation of Ca2.handling proteins on the Ca2.transients at early,intermediate and later differentiation stages of ES cell-derived cardiomyocytes(ESCM).Results:Nifedipine,a selective antagonist of L-type Ca2.channels,totally blocked Ca2.transients even in the condition of field-electric stimulation inESCM at three differentiation *** Ca2.transients of ESCM were alsoinhibited by both ryanodine[an inhibitor of ryanodine receptors(RyRs)]and 2.aminoethoxydipheylborate[2.APB,an inhibitor of inositol-1,4,5-trisphosphatereceptors(IP3Rs)].The inhibitory effect of ryanodine increased with the time ofdifferentiation,while the effect of 2.APB decreased with the ***,an inhibitor of SR Ca2.-pump ATPase,inhibited Ca2.transientsequally at three differentiation stages that matched the expression ***~+free solution,which inhibits Na~+-Ca2.exchanger(NCX)to extrude Ca2.from cytosol,did not affect the amplitude of Ca2.transients of ESCM until the latter differentia-tion stage,but it significantly enhanced the basal Ca2.***:The Ca2.transients in ESCM depend on both the sarcolemmal Ca2.entry viaL-type Ca2.channels and the SR Ca2.release from RyRs and IP3Rs even at theearly differentiation stage;but NCX seems not to regulate the peak of Ca2.tran-sients until the latter differentiation stage.

关键词： Ca2. transients cardiac differentiation embryonic stem cells Ca2. handing proteins

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人纤溶酶原K5区基因真核表达载体的构建和活性鉴定

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复旦学报（医学版） 2003年第5期30卷 409-413,F002页

作者：杨健王跃祥官孝群马春姑陶贤梅宋后燕复旦大学上海医学院细胞与遗传学系-教育部分子医学重点实验室上海200032

目的　构建含人纤溶酶原K5区基因的真核表达载体 ,转染人乳腺癌细胞株MDA MB 2.31,观察阳性克隆表达的K5蛋白对人脐静脉内皮细胞株ECV30 4和MDA MB 2.31细胞增殖的影响。方法　应用PCR将人纤溶酶原信号肽序列引入K5cDNA ,所得的目的片... 详细信息

目的　构建含人纤溶酶原K5区基因的真核表达载体 ,转染人乳腺癌细胞株MDA MB 2.31,观察阳性克隆表达的K5蛋白对人脐静脉内皮细胞株ECV30 4和MDA MB 2.31细胞增殖的影响。方法　应用PCR将人纤溶酶原信号肽序列引入K5cDNA ,所得的目的片段与真核表达载体 pcDNA3重组 ,构建重组质粒pcDNA3K5 ,脂质体法将其转染MDA MB 2.31,G4 18筛选阳性克隆 ,PCR鉴定 ,RT PCR和Westernblot检测K5的表达。将鉴定正确的阳性克隆的培养上清作用于ECV30 4细胞 ,MTT法检测其增殖情况 ,并用MTT法检测转染 pcD NA3K5对MDA MB 2.31细胞增殖的影响。结果　构建的重组质粒 pcDNA3K5经酶切鉴定、测序正确 ,将其转染MDA MB 2.31后挑取的阳性克隆有 3个经PCR鉴定正确 ,并经RT PCR和Westernblot检测证实K5的表达。阳性克隆的培养上清作用于ECV30 4细胞后 ,其存活率降低 ;转染 pcDNA3K5对MDA MB 2.31细胞增殖无明显影响。结论　应用脂质体法将带有人纤溶酶原信号肽序列的K5cDNA转染MDA MB 2.31细胞后 ,其分泌产生的有生物学活性的K5 ,呈现对ECV30 4细胞增殖的抑制作用 ,而对MDA MB 2.31细胞的生长则无影响。

关键词：乳腺癌基因治疗重组新生血管生成抑制因子K5 基因表达真核表达载体构建活性鉴定

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Fabrication of gelatin-hyaluronic acid hybrid scaffolds with tunable porous structures for soft tissue engineering

Fabrication of gelatin-hyaluronic acid hybrid scaffolds with...

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生物材料与再生医学的现状与未来——2.10年第十届上海地区医用生物材料研讨会

作者： Fan Zhang~2.Chuanglong He~(1,2.3*),Lijun Cao~2.Wei Feng~2.Hongsheng Wang~2.Xiumei Mo~(1,2.3) Jinwu Wang~(4*) 1 key laboratory of Textile Science & Technology,{2. of {2.,Donghua University,shanghai,P.R. china 2.162. 2.College of Chemistry,Chemical Engineering and Biotechnology,Donghua University,shanghai,P.R.china 2.162. 3 State key laboratory for Modification of Chemical Fibers and Polymer Materials,Donghua University, shanghai,P.R.china 2.162. 4 Department of Orthopaedic Surgery,Ninth People’s Hospital,shanghai Jiao Tong University School of medicine. shanghai,P.R.china 2.0011

The development of three -dimensional(3 -D) scaffolds with highly open porous structure is one of the most important issues in tissue *** this study,3-D macroporous gelatin/hyaluronic acid(GE/HA) hybrid scaffolds with varying porous morphology were prepared by freeze -drying their blending solutions and subsequent chemical crosslinking by using 1 -Ethyl -3 -(3 -dimethylaminopropyl) carbodiimide hydrochloride (EDC).The resulting scaffolds were characterized by scanning electron microscopy(SEM) and Fourier transform infrared spectroscopy(FTIR).Their swelling,in vitro degradation properties and compressive strength were also *** evaluate in vitro cytocompatibility of scaffolds,mouse L92. fibroblasts were seeded onto the scaffolds for cell morphology and cell viability *** was found that the porous structure of scaffolds can be tailored by varying the ratios of gelatin to HA,both the swelling ratios and degradation rate increased with the increase of HA content in hybrid scaffolds,and crosslinking the scaffolds with EDC improved the degradation resistance of the scaffold in culture media and increased the mechanical strength of *** in vitro results revealed that the prepared scaffolds do not induce cytotoxic effects and suitable for cell growth,especially in the case of scaffolds with higher gelatin *** combined results of the physicochemical and biological studies suggested that the developed GE/HA hybrid scaffolds exhibit good potential and biocompatibility for soft tissue engineering applications.

关键词： Gelatin Hyaluronic acid Porous scaffold Freeze-drying Tissue engineering

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Activation of alphal-adrenergic receptors upregulates the miR-199a by serum response factor in cardiomyocytes

Activation of alphal-adrenergic receptors upregulates the mi...

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第七届海峡两岸心血管科学研讨会

作者： Wei Feng,Youyi Zhang Institute of Vascular medicine.Peking University the Third Hospital and key {2. of {2. Cardiovascular Science, ministry of education,Beijing 100191,P.R.china

＜正＞MicroRNA miR-199a,which was expressed in heart,was modulated in hypertrophic or failing hearts from rodent and *** study was aimed to determine whether alpha1-adrenergic receptor（alpha1-AR）stimulated by phenylephrine regulates miR-199a expression in rat hearts and,if so,to examine the underlying *** found that incubation of neonatal rat cardiomyocytes with phenylephrine（1μmol/L and 10μmol/L for 2. h）increased mature miR-199a expression（1.32.0.11 and 1.61±0.2. fold of control respectively）and enhanced miR-199a promoter activity, which was fully abolished by a alpha1-AR antagonist,prezosin（1 mol/L）.Further promoter assays by deletion and mutation of the 5’ upstream region of miR-199a promoter showed that serum response element（SRE）was critical for miR-199a *** of SRF resulted in a strong stimulation of miR-199a promoter activity about 3.2.±0.2. *** the transcription factor serum response factor（SRF）binding to miR-199a promoter was increased after phenylephrine simulation confirming by chromatin *** conclusion,the SRF was the downstream transcriptional mediator of phenylephrine-induced miR-199a *** miR-199a is controlled by SRF via a SRE and might play a role as a novel intermediate in alpha1-adrenergic effects in the heart.

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New Strategy to Construct Fused/Bridged/Spiro Carbocyclic Scaffolds Based on the Design of Novel 6-C Synthon Precursor

New Strategy to Construct Fused/Bridged/Spiro Carbocyclic Sc...

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第十六届全国金属有机化学学术讨论会

作者： Jia Liu,~(1,2. Xi Wang,~1 Chang-Liang Sun,~1 Bi-Jie Li,~1 Min Wang,~2.Zhang-Jie Shi~(*1,3) 1 Beijing National laboratory of molecular Sciences(BNLMS) and {2. laboratory of Bioorganic Chemistry and molecular Engineering of ministry of education,College of Chemistry and molecular Engineering and Green Chemistry Center,Peking University,Beijing 100871 and Department of Chemistry,2.University of Agriculture of china,Beijing 100094 and 3 State {2. laboratory of Organometallic Chemistry,shanghai Institute of Organic Chemistry,Chinese Academy of Sciences,shanghai 2.0032.china

＜正＞Fused/bridged/spiro carbocyclic ring scaffolds are one of most important motifs in natural and synthetic organic *** design a new 6-C synthon,which could be applied to construct complex poly-,spiro-and bridged systems through the simple sequential chemical transformations from acyclic precursor by a one-pot strategy from

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D2. like but not D1- like dopamine receptors in ventrolateral orbital cortex are involved in apomorphine-induced antiallodynia in a neuropathic pain model of rats

D2- like but not D1- like dopamine receptors in ventrolatera...

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中国神经科学学会第四次会员代表大会暨第八届全国学术会议

作者： Yong-Hui DANG~1,Yan ZHAO~1,Bo XING~1,Xiao-Jie ZHAO~1,Teng CHEN~1,Jing-Shi TANG~2.Fu-Quan HUO~2.Feng-Yi XU~2. Chao-LingQU~(2.*) 1 Department of Forensic medicine.key {2. of Environment and Gene Related to Diseases,{2. of education, Xi''an Jiaotong University School of medicine.Xi''an 710061,china 2.Department of Physiology and Pathophysiology,key {2. of Environment and Genes Related to Diseases, {2. of education,Xi''an Jiaotong University School of medicine.Xi''an 710061,china

＜正＞Objective The ventrolateral orbital cortex（VLO） is part of an endogenous analgesic system consisting of an ascending pathway from the spinal cord to VLO via the thalamic nucleus submedius（Sm） and a descending pathway to the spinal cord relaying in the periaqueductal gray（PAG）.The aim of the present study was to examine whether dopamine and its receptors are involved in the antinociception in *** The animal model of persistent peripheral neuropathic pain-spared nerve injury（SNI） was *** withdrawal threshold in response to von-Frey hair stimulation of the injury side hindpaw was measured as an index of mechanical ***（1） Microinjection of the nonselective dopamine receptor agonist apomorphine（1.0,2.5,5.0μg in 0.5μl） into the VLO significantly increased the paw withdrawal threshold in a dose-dependent manner,which was reversed by pretreatment with selective D2.like dopamine receptor antagonist raclopride（1.5μg in 0.5μl）,but not selective D1-like dopamine receptor antagonist（SCH2.390 5.0μg in 0.5μl）.（2. Microinjection of raclopride or SCH2.390 alone into the VLO had no effects on mechanical paw withdrawal threshold. Conclusion The results suggest that dopamine neurotransmitter is involved in VLO-induced anti-allodynia,and this effect is mediated by D2.like but not Dl-like dopamine receptors.

关键词： ventrolateral orbital cortex dopamine receptor raclopride SCH2.390 mechanical allodynia spared nerve injury rat

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生长激素相关基因突变与原发闭经的关系

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华中科技大学学报（医学版） 2005年第4期34卷 472-474页

作者：刘学飞杨真荣王慧周波陈燕唐艳平复旦大学上海医学院分子医学教育部重点实验室上海200032 华中科技大学同济医学院遗传学教研室武汉430030

目的探讨生长激素相关基因突变与原发闭经的关系。方法对130例确诊为原发闭经的患者和30名健康女性个体进行核型分析,并采用单链构象多态性分析(SSCP)等方法,对原发闭经患者的生长激素释放激素受体基因(GHRHR)进行检测。结果有33例原发... 详细信息

目的探讨生长激素相关基因突变与原发闭经的关系。方法对130例确诊为原发闭经的患者和30名健康女性个体进行核型分析,并采用单链构象多态性分析(SSCP)等方法,对原发闭经患者的生长激素释放激素受体基因(GHRHR)进行检测。结果有33例原发闭经患者出现染色体核型异常,1例出现GHRHR基因点突变,且该突变位点在第7个外显子中。结论GHRHR基因第7个外显子的点突变可能是原发闭经的病因之一。

关键词：原发闭经单链构象多态性分析生长激素生长激素释放激素受体

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Understanding structure and function of acid-sensing ion channels

Understanding structure and function of acid-sensing ion cha...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Tian-Le XU~(1,2. 1 Institute of Neuroscience and State key laboratory of Neuroscience,{2. Institutes for Biological Sciences, Chinese Academy of Sciences,{2. 2.0031,china 2.Neuroscience Division,Department of Biochemistry and molecular Cell Biology,Institute of Medical Sciences, {2. Jiaotong University School of medicine.{2. 2.002.,china

The acid-sensing ion channel(ASIC) has a prominent role in tissue pH-sensing and neuronal signalling. Subunits of ASICs are members of the degenerin/epithelial Na channel protein *** date,seven ASIC subunits have been identified:ASIC1a,ASIC1b,ASIC1b2.ASIC2.,ASIC2.,ASIC3,and *** subunit contains cytosolic N and C termini,two alpha-helical transmembrane domains(M1-M2.,and a disulfide-rich multidomain extracellular *** form a variety of homo- and heterotrimeric ion channels that are activated by extracellular acidosis associated with neurotransmission,injury,inflammation and ischemia,triggering membrane depolarization via Na influx into target *** the operating mechanisms of these ion channels could therefore provide the catalyst for further studying ASIC physiology and pharmacology,as well as promoting the development of experimental techniques for other families of ion channels and/or *** this talk,I will present our current work on ASIC structure and function,in particular those of ASIC1a and ASIC3 *** in this area has been stimulated in recent years by the publication of high-resolution structures of ASIC1,which have permitted the structure-directed approaches and the creation of ASIC homology models.I will first discuss the crucial roles of ASICs in mediating neuronal injury and pain *** a series of computational,chemical and functional approaches,and nonproton ligands(GMQ,agmatine and amiloride) as probes,I will then delineate the nonproton ligand sensing domain in ASIC3 channels at the atomic level and describe how chemical biology,homology modeling and ligand docking can provide useful information about ligand interactions.I will also consider some of the ongoing projects in my laboratory that are investigating the pathophysiological roles of ASICs using ASIC subunit gene knockout *** report ends with a discussion on the generality of ASICs as a model system for the study of struc

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Switching of N-methyl-D-aspartate(NMDA) receptor-favorite intracellular signal pathways from ERK1/2.to p38 MAPK leads to developmental changes in NMDA neurotoxicity

Switching of N-methyl-D-aspartate(NMDA) receptor-favorite in...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Lin XIAO~*,Chun HU,Chun-Zhi FENG,Yi-Zhang CHEN~* Institute of Neuroscience,key laboratory of {2. neurobiology,{2. of education,Second Military Medical University,shanghai 2.0433,china

Excitotoxicity mediated by overactivation of N-methyl-D-aspartate receptors(NMDARs) has been implicated in a variety of neuropathological conditions in the central nervous system(CNS).It has been suggested that NMDA neurotoxicity is developmentally regulated,but the definite pattern of the regulation has been controversial and the underlying mechanism remains largely ***,we show that NMDA treatment leads to significant cell death in mature(9,12.IV) hippocampal neurons or hippocampi of young(P12. and adult rats but not in immature (3,6DIV) neurons or embryonic(E18) and neonatal rat *** contrast,NMDA promotes survival of immature neuron against tropic ***,it is found that NMDA preferentially activates p38 mitogen activated protein kianse(MAPK) in mature neuron and adult rat hippocampus,but favors extracellular signal-regulated kinases1/2.ERK1/2. activation in immature neuron and P0 rat ***,it is shown that NMDA neurotoxicity in mature neuron is mediated via p38 MAPK activation,and neuroprotection in immature neuron is mediated via ERK1/2.activation,while all these effects are NR2.-containing NMDARs dependent,as well as Ca *** also revealed that mature and immature neuron showed no difference in the amplitude of NMDAinduced intracellular calcium([Ca]) ***,the basal level of[Ca] is shown to elevate with the maturation of neuron,and this elevation is attributable to the changes in NMDA neurotoxicity but not to the switch of NMDAR signaling *** together,our results suggest that a switch of NMDA receptor-favorite intracellular signal pathways from ERK1/2.to p38 MAPK and the elevated basal level of[Ca] with age might be critical for the developmental changes in NMDA neurotoxicity in hippocampal neuron.

关键词： NMDA receptor neurotoxicity development MAPK calcium

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Nongenomic effects of corticosterone on NMDA receptor-mediated current in cultured embryonic 18d rat hippocampal neurons

Nongenomic effects of corticosterone on NMDA receptor-mediat...

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中国神经科学学会第四次会员代表大会暨第八届全国学术会议

作者： Yan-Min ZHANG,Hui SHENG,Yan-Ming CHENG,Xin NI* Department of Physiology,the key laboratory of {2. Neurobiology,{2. of education,the Second Military Medical University,shanghai 2.0433,china

Objective Glucocorticoids(Gc),main stress hormone,has been shown to modulate synaptic plasticity and the process of learning and memory in *** exerts long-term effects on central nervous system(CNS) through classical *** is increasing evidence for rapid Gc actions triggered by the activation of membrane-associated receptors and nongenomic signaling *** N-mentyl-D-aspartate(NMDA) receptor plays a pivotal role in a variety of physiological processes in the CNS,including LTP,with involvement in learning and ***, nongenomic effects of Gc on N-mentyl-D-aspartate(NMDA) receptor,the key receptor for synaptic plasticity,remain *** Whole-cell patch-clamp was performed on primary cultured E18 hippocampal *** PLC-β3 was measured by Western *** cAMP content was measured by RIA ***(1) Primary cultured embryonic hippocampal neurons were deficient in glucocorticoid *** corticosterone(B)(10-7 mol/L) and BSA-conjugated corticosterone(B-BSA)(10-7 mol/L) elicited a rapid depression of NMDA-induced currents in a dose-dependent *** effect was reversed by the G-protein antagonist(GDP-β-S),but not by glucocorticoid receptor antagonist(Ru38486) or mineralocorticoid receptor antagonist(spironolactone).(2. Investigations on the signaling pathways of B showed that B dose-dependently induced phosphorylated PLC-βexpression and increased intracellular cAMP content in these cells.(3) Blocking protein kinase A(H89) and adenylate cyclase(SQ 2.536) partly abolished the Binduced depression of NMDA current.(4) Application of inhibitor of phospholipase C(U7312.),IP3 receptor antagonist (2.PB),Ca chelator(BAPTA) and protein kinase C inhibitor(chelerythrine) prevented B-induced depression of NMDA current.(5) Application of inhibitors of JNK(SP60012.) and p38 MAPKs(SB2.2.90) prevented B-induced depression of NMDA current,whereas blocking ERK activity failed to do the *** results sug

关键词： stress corticosterone N-mentyl-D-aspartate receptor nongenomic

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