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Effects of disinhibition on spatiotemporal pattern of neuronal first recruitment in neuronal networks

Progress in Natural Science:Materials International 2009年第5期19卷 615-621页

作者： Liangbin Pana,b,c,1, Xindong Songa,b,c,1, Guangxin Xiang a,b,c, Jing Zhua,b,c, Jing Cheng a,b,c,d,a Medical Systems Biology Research Center, tsinghua university School of Medicine, beijing 100084,{2. b department of biological Sciences and biotechnology, tsinghua university, beijing 100084,{2. c National Engineering Research Center for beijing Biochip Technology, 18 Life Science Parkway, beijing 102.06,{2. d {{12. Key laboratory of biomembrane and Membrane biotechnology, tsinghua university, beijing 100084,{2. [a]Medical Systems Biology Research Center Tsinghua University School of Medicine Beijing 100084 China [b]Department of Biological Sciences and Biotechnology Tsinghua University Beijing 100084 China [c]National Engineering Research Center for Beijing Biochip Technology 18 Life Science Parkway Beijing 102206 China [d]State Key Laboratory of Biomembrane and Membrane Biotechnology Tsinghua University Beijing 100084 China

The propagation of neuronal activities is a key feature to understanding information processing in networks. The analysis based on first-spikes of bursts in turn plays an important role in the research of neuronal activity propagation. Our focus here is to investigate how spatiotemporal patterns of neuronal first-spikes are affected by disinhibition. Multi-electrode arrays were used to record stimulation-evoked bursts of multiple neurons in randomly cultured neuronal networks. Both the precise timing of and the rank relationships between first-spikes were analyzed. Compared with evoked bursts in the network’s native state, the precise first-spike latencies in its dis-inhibited state are more consistent and the propagation of its bursting activities is much faster. Additional points of interest are that disinhibited neuronal networks can be evoked to generate stable and distinguishable neuronal first recruitment spatiotemporal patterns specific to the stimulation site, and that the disinhibition may cause the original spatiotemporal patterns to change in a heterogeneous manner with regards to different propagation pathways.

关键词： Neuronal recruitment Spatiotemporal pattern Disinhibition Network Multi-electrode array

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Cloning and sequence characteristics of the genomic gene of a rice metallothionein

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Chinese Science Bulletin 2000年第2期45卷 153-156页

作者： YU Lihua, LIU Jinyuan, Masaaki Umeda, Hirofumi Uchimiya & ZHAO Nanming department of biological Sciences and {2., state Key laboratory of biomembrane and Membrane {2., beijing 100084, china Institute of Molecular and Cellular Bioscie department of biological Sciences and {2. State Key Laboratory of Biomembrane and Membrane Biotechnology Beijing China Institute of Molecular and Cellular Biosciences The University of Tokyo Tokyo Japan

Northern blot analysis showed that a metallothionein gene, ricMT is expressed strongly in the stem of rice with an expression level that could be more than 100-fold stronger than in leaf blades. The results suggest that the 5’ upstream region flanking the coding sequence of the ricMT may contain a fairly strong promoter To elucidatr its regulation and promoter structure, the genomic clones of ricMT were screened out from a rice genomic library and a fragment of about 4084 bp was sequenced. The fragment included a 5’ upstream region of ca. 2.70 bp, a transcription region of ca. 690 bp and a 3’ downstream region of ca. 42. bp. Computer analysis of the sequence homology showed that the 5’ upstream region included a putative TATA box, a putative CAAT box, and a typical metal-responsive element TGCGCGCG. The results will promote further understanding of the mechanisms of gene regulation and metal response of plant metallothionein proteins.

关键词： rice metallothionein genomic gene regulatory sequence.

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The excitation energy transfer between β-Car and Chla molecules in PS Ⅱ core antenna complexes CP43 and CP47

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Chinese Science Bulletin 2000年第17期45卷 1579-1583页

作者： SHAN Jixiu, WANG Jushuo, LI Liangbi,ZHAO Nanming & KUANG Tingyun1. laboratory of Photosynthesis Basic Research, Institute of Botany, Chinese Academy of Sciences, beijing 100093,{2. 2. department of biological Sciences and biotechnology, state Key laboratory of biomembrane and Membrane biotechnology, tsinghua university, beijing 100084,{2.Correspondence should be addressed to Kuang Tingyun laboratory of Photosynthesis Basic Research Institute of Botany Chinese Academy of Sciences Beijing China department of biological Sciences and {2. State Key Laboratory of Biomembrane and Membrane Biotechnology Tsinghua University Beijing China

The excitation energy transfer efficiency between p-Car and Chla molecules in purified CP43 and CP47 was calculated by comparing absorption and fluorescence excitation after normalization at 550 nm, CP43 had an energy transfer efficiency of 2..1% while the CP47 had an energy transfer efficiency of 62.8%, proving that excitation energy was transferred between p-Car and Chla molecules in CP43 and CP47 at normal conditions. The excitation energy transfer between p-Car and Chla molecules in CP43 and CP47 may occur through the "Dexter" mechanism and the distance between these two kinds of pigments should be less than 1 nm. In addition, the results were also used to discuss the conformational relationship between p-Car and Chla molecules in CP43 and CP47.

关键词： CP43 CP47 β-Car Chla excitation energy transfer.

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突触结合蛋白Ⅰ的胞质片段与磷脂膜的相互作用

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生物化学与生物物理进展 2003年第1期30卷 84-88页

作者：贺雨虹隋森芳清华大学生物科学与技术系

突触结合蛋白Ⅰ是神经细胞突触囊泡上的一个膜整合蛋白 ,C2.B是其具有重要功能的胞质片段 .近年的研究表明 ,突触结合蛋白Ⅰ在钙引发的神经递质快速释放过程中起到钙感受器的作用 ,它与神经细胞突触前膜的相互作用与其生理功能有关 ,但... 详细信息

突触结合蛋白Ⅰ是神经细胞突触囊泡上的一个膜整合蛋白 ,C2.B是其具有重要功能的胞质片段 .近年的研究表明 ,突触结合蛋白Ⅰ在钙引发的神经递质快速释放过程中起到钙感受器的作用 ,它与神经细胞突触前膜的相互作用与其生理功能有关 ,但是其作用机制还不清楚 .利用气 /液单层膜技术结合荧光发射光谱和圆二色光谱技术 ,发现C2.B倾向于插入带负电荷的磷脂膜中 (如磷脂酰丝氨酸 ) ,而且插膜是钙依赖性的 ;对于不带电荷的磷脂不插膜 .C2.B与膜之间的作用力主要为静电力 .荧光发射光谱和圆二色光谱结果显示 ,它与膜相互作用时二级结构不发生显著变化 .结果表明 ,突触结合蛋白Ⅰ钙依赖的插入负电荷膜特点 ,可以帮助解释其钙感受器的作用机制 .

关键词：突触结合蛋白I 胞质片段磷脂膜相互作用临界插膜压神经细胞

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In vitro and in vivo evaluation of a novel oral insulin formulation

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Acta Pharmacologica Sinica 2006年第10期27卷 1382-1388页

作者： Er-li MA Hong MA Zheng LIU Chang-xue ZHENG Ming-xing DUAN~2.state Key laboratory of Bio-membrane and {2.,department of biological Sciences and {2.,tsinghua university beijing 100084,china state Key laboratory of Bio-membrane and {2. Department of Biological Sciences and Biotechnology Tsinghua University Beijing 100084 China

Aim:To develop a stable self-emulsifying formulation for oral delivery of ***:Caco-2.cell line and diabetic beagles were used as in vitro and in vivomodels to study the absorption mechanism and the hypoglycemic efficacy of *** addition,various physicochemical parameters of the formulationsuch as droplet size,insulin encapsulation efficiency and stability were ***:This formulation enabled changes in barrier properties of Caco-2.onolayers,as referred by transepithelial electrical resistance(TEER)and appar-ent permeability coefficients(Papp)of the paracellular marker ranitidine(2.-foldgreater than control)but not transcellular marker propranolol,suggesting that theopening of tight junctions was *** diabetic beagle dogs,the bioavailabilityof this formulation was up to 15.2. at a dose of 2.5 IU/kg in comparison with thehypoglycemic effect of native insulin(0.5 IU/kg)delivered by ***:This formulation,recently approved by the china stateFood and Drug Administration to enter clinical trials,was stable,degradation-protected and absorption-enhanced,and provided a promising formulation fororal insulin delivery.

关键词： insulin oral delivery Caco-2 diabetic beagles

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MEK inhibitor PD98059 acutely inhibits synchronized spontaneous Ca2.oscillations in cultured hippocampal networks

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Acta Pharmacologica Sinica 2006年第7期27卷 869-876页

作者： Yan-fang RUI~1 Zhao-hui SUN~2.Jia-ping GU~1 Zhong-hua SHENG~1 Xiang-ping HE~1 Zuo-ping XIE~(1,3)1 department of biological Science and {2.,state Key laboratory of biomembrane and Membrane {2.,tsinghua university beijing 100084,china 2.Institute of Neurological Disorders,Life Science and Medical department,tsinghua university,beijing 100084,china departmentofbiologicalScienceand{2. StateKeyLaboratoryofBiomembraneandMembraneBiotechnologyTsinghuaUniversityBeijing100084China InstituteofNeurologicalDisorders LifeScienceandMedicalDepartment

Aim:To investigate the changes in synchronized spontaneous Ca2.oscillationsinduced by mitogen-activated protein kinase kinase（MEK）inhibitor PD98059 atdifferent concentrations in cultured hippocampal ***:Hippocam-pal neurons in culture for 1-2.weeks were used for this *** activities of these hippocampal neurons were examined by Ca2.imagingusing calcium-sensitive *** inhibitor PD98059（10,30,and 60μmol/L）andSB2.2.74（10 and 60μmol/L）,a negative control for mitogen-activated proteinkinase（MAPK）cascade study,were applied to the cells under the microscopewhile imaging was taking ***:PD98059 at a lower concentration of 10μmol/L had little effect on the Ca2.*** the higher concentration of 30μmol/L,5 min after application of PD98059,the spike frequency was decreased to2..38%±7.40%（mean±SEM,n=16,P2.2.74 atconcentrations of 10 and 60 μmol/L had little inhibition effect on the Ca2.***:These results indicate that PD98059 inhibits synchronized sponta-neous Ca2.oscillation through inhibition of MEK,which hints that the MAPKcascade is required to maintain synchronized spontaneous Ca2.oscillation.

关键词： PD98059 MEK calcium oscillation

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表面等离激元共振生物传感器研究整合素蛋白α_(Ⅱb) β_3与RGD多肽的特异结合

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生物化学与生物物理进展 2002年第5期29卷 796-800页

作者：路英杰张帆隋森芳清华大学生物科学与技术系生物膜与膜生物工程国家重点实验室北京100084

整合素蛋白αⅡbβ3 是血小板上的一种钙依赖性膜受体 ,其胞外结构域可与含有RGD序列的肽链特异结合 .通过将含有NTA DOGS的磷脂单分子层膜转移到 5 0nm厚的金膜上 ,制备了一种含有NTA头部的表面等离激元共振 (SPR)传感器敏感膜 .设计... 详细信息

整合素蛋白αⅡbβ3 是血小板上的一种钙依赖性膜受体 ,其胞外结构域可与含有RGD序列的肽链特异结合 .通过将含有NTA DOGS的磷脂单分子层膜转移到 5 0nm厚的金膜上 ,制备了一种含有NTA头部的表面等离激元共振 (SPR)传感器敏感膜 .设计并合成了含有 6个组氨酸和RGD基团的His tagged短肽P1,并利用SPR生物传感器 ,对整合素蛋白与含有RGD配基的支撑平面膜的特异相互作用以及Ca2.+ 、Mn2.+ 对该相互作用的影响进行了研究 .结果表明 ,NTA敏感膜能很好地将P1锚定在支撑平面膜表面 ,并能够保证P1维持一个有效的定向 .将Ca2.+ 从低结合位点去除或加入Mn2.+ 都能够增加整合素蛋白的配基结合活性 .二价阳离子对整合素蛋白配基结合能力的调节作用。

关键词：表面等离激元共振生物传感器整合素蛋白 αⅡbβ3 RGD多肽特异结合

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PTEN,a general negative regulator of cyclin D expression

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Cell Research 2007年第4期17卷 291-292页

作者： Eirong Diao~1 Ye-Guang Chen ~1state Key laboratory of {2. and Membrane biotechnology,department of biological Sciences and biotechnology Tsing-hua Universiry,beijing 100084,china stateKeylaboratoryof{2.andMembranebiotechnology DepartmentofBiologicalSciencesandBiotechnologyTsing-huaUniversityBeijing100084China

The tumor-suppressor phosphatase with tensin homol-ogy(PTEN)is frequently mutated in many malignanciesand is one of the most well studied tumor suppressor genes[1,2..PTEN,a lipid and protein dual phosphatase,plays avital role in embryonic development,cell growth,apoptosisand cell *** well-known function of PTEN isphosphatidylinositol-3(PI)-phosphatase,which functionsas a negative regulator of the PIkinase(PI3K)*** well established that PTEN regulates the G1-S transitionby modulating the expression of cyclin D1 and *** passage of any cell through cell cycles is subtlymodulated by a series of proteins,among which the mainplayers in animal cells include cyclins,CDKs(cyclin-de-pendent kinases,positive regulators)and CDK inhibitors(negative regulators such as p2.and *** Dsare key regulators during the G1 phase and there are threetypes of cyclin Ds,D1,D2.and D3 in mammalian cells[3].Although the phenotypic analyses of single cyclin Dknockout mice revealed that each of the D type cyclinsis sufficient to drive normal development of the majority

关键词： PTEN,a general negative regulator of cyclin D expression

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Cloning, Expression, Purification and Crystallization of NHR3 Domain from Acute Myelogenous Leukemia-related Protein AML1-ETO

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Acta Biochimica et Biophysica Sinica 2004年第8期36卷 566-570页

作者： Hai-Tao YANG , Dong-Hui WU1 , Xiao-Yu XUE, Wen-Xue LIANG1, Xiao-Yu MIAO,Hai PANG , and Sai-Juan CHEN1laboratory of Structural Biology, department of {2. Sciences and biotechnology & Protein Sciences laboratory of MOE, tsinghua university, beijing 100084, china Shanghai Institute of Hematology, state Key laboratory of Medical Genomics,1 Shanghai Second Medical university, Shanghai 2.002., china laboratory of Structural Biology Department of Biological Sciences and Biotechnology & Protein Sciences Laboratory of MOE Tsinghua University Shanghai Institute of Hematology State Key Laboratory of Medical Genomics Shanghai Second Medical University

The t(8;2.) translocation is one of the most frequent chromosome abnormalities in acute myeloid leukemia. This translocation creates a fusion between the acute myelogenous leukemia 1 (AML1, a transcription factor) gene on chromosome 2. and the eight-twenty-one (ETO, a zinc finger nuclear protein) gene on chromosome 8, leading to the repression of certain AML1 target genes. We cloned NHR3 domain coding fragment into vector pGEX-6p-1 using PCR and obtained recombinant plasmid pGEX-6p-1-NHR3, which can be induced to stably overexpress fusion protein in E. coli. Through the purification on GST affinity chromatography column and PreScission protease cleavage, a large amount of NHR3 protein with high purity was obtained. In order to avoid possible interference of some strong negative charged molecules, NHR3 protein was further purified by Mono Q anion exchange chromatography. The NHR3 crystals were grown with hanging drop/vapor diffusion method and the first crystals appeared after four weeks at 18 °C in 0.2.M Tris-sodium citrate dihydrate, 0.1 M sodium cacodylate, pH 6.5, and 30% iso-propanol (V/V). ESI mass spectrum showed that the molecular weight of this domain was in agreement with its primary structure sequence prediction, and circular dichroism spectral data (190–2.0 nm) showed that NHR3 had a well-defined secondary structure of 2..9% α-helix, 2..2. random coil and 50.9% turn, which was consistent with GOV4 software prediction.

关键词： AML1-ETO NHR3 crystallization ESI mass spectrum circular dichroism spectrum glutathione S-transferase (GST)

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禽流感及其免疫防制研究

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生物化学与生物物理进展 2004年第3期31卷 189-197页

作者：段明星何宏轩张强哲清华大学生物科学与技术系

禽流感一直是严重危害世界各国养禽业发展的头号大敌 ,近期又成为继严重急性呼吸道综合症 (SARS)后又一严重威胁人类生命安全的重要疾病 ,由于禽流感病毒抗原及其致病力的易变性 ,这就要求未来的防制策略要采取快速检测 ,并用先进的分... 详细信息

禽流感一直是严重危害世界各国养禽业发展的头号大敌 ,近期又成为继严重急性呼吸道综合症 (SARS)后又一严重威胁人类生命安全的重要疾病 ,由于禽流感病毒抗原及其致病力的易变性 ,这就要求未来的防制策略要采取快速检测 ,并用先进的分子生物学技术进行病毒鉴定、检疫及免疫保护等措施 ,建立全国性甚至全球性禽流感检测防制网络 ,并且搞清禽流感与人类流感的关系 ,从而保证人和动物的安全 .

关键词：禽流感人类分子生物学技术病毒鉴定检疫免疫保护防治禽类流行性感冒

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