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A patch-clamp study on human sperm Cl^- channel reassembled into giant liposome

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Asian Journal of Andrology 2001年第3期3卷 185-191页

作者： Jun-Ping BAI yu-liang shi Key Laboratory of Neurobiology,Intitute of physiology,shanghai institutes for biological sciences,chinese academy of Science, shanghai 200031,china Key Laboratory of Neurobiology Institute of Physiology Chinese Academy of Sciences Shanghai.

Aim: To record the single-channel currents and characterize the electrophysiological properties of the Cl^- channels inhuman sperm membrane. Methods: The membrane proteins extracted from the human sperm were reassembled intoliposome bilayer, and the liposomes were fused into giant liposomes with a diameter more than 10μm by dehydration-rehydration procedure. The giant liposomes were used to study the Cl^- channel activities by patch-clamp ***: By patch clamping the giant liposome in an asymmetric NMDG (N-methyl-D-glucamine)-Cl (bath 100//pipette 200 mmol/L) solution system, three kinds of single-channel events with unit conductances of (74.1 ± 8.3) pS,(117.0±5.7) pS and (144.7±4.5) pS, respectively, were detected. Their activities were voltage-dependent and allwere blocked by SITS (4-acetamido-4'-isothiocyanato-stilbene-2', 2'-disulfonic acid) in a concentration-dependentmanner. By constructing the open and close dwell time distribution histograms and then fitting them with exponentialfunction, two time constants were obtained in both the open and the close states. The burst activity and conductancesubstate of the channels were observed. Conclusion; There exist three kinds of Cl^- channels with different conduc-tance in human sperm membrane at least. (Asian J Androl 2001 Sep; 3: 185 - 191)

关键词： Cl^-channel giant liposome patch clamp human sperm membrane ion channel reassembly

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Identification of connexin 50 and 57 mRNA in A-type horizontal cells of the rabbit retina

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Cell Research 2005年第3期15卷 207-211页

作者： He HUANG*,**,Hui li*,shi Gang HE institute of Neuroscience,shanghai institutes for biological sciences,chinese academy of sciences,320 yueyang Road,shanghai 200031,china institute of Neuroscience Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China

Horizontal cells (HCs) mediate negative feedback to photoreceptors. In the mammalian retina, there are two types of HCs, which are extensively coupled to neighboring cells through homologous gap junctions. The permeability and therefore the strength of feedback can be regulated by light intensity, dopamine and many other factors. However, the component(s) of the most prominent gap junctions, those between A-type HCs in the rabbit retina, is still unknown. In this study, we compared the sequences of many types of mammalian connexins, obtained partial sequences of rabbit connexin 50 and 57. Using specific primers designed against the rabbit sequences, we identified mRNAs of connexin 50 and/or 57 in visually selected single A-type HC using multiplex RT-PCR.

关键词： gap junctions dissociated retinal neurons single cell multiplex RT-PCR sequencing.

In Vitro Reassembly of Tobacco Ribulose-1,5-bisphosphate Carboxylase/Oxygenase from Fully Denatured Subunits

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Acta Biochimica et Biophysica Sinica 2006年第10期38卷 737-745页

作者： Zhen-Hua YONG Gen-yun CHEN* Jiao-Nai shi Da-Quan XU institute of Plant physiology and Ecology,shanghai institutes for biological sciences,chinese academy of sciences,and Graduate School of the chinese academy of sciences,shanghai 200032,china institute of Plant physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences and Graduate School of the Chinese Academy of SciencesShanghai 200032 China

It has been generally proved impossible to reassemble ribulose-1,5-bisphosphate carboxylase/oxygenase （Rubisco） from fully denatured subunits in vitro in higher plant,because large subunit of fullydenatured Rubisco is liable to precipitate when the denaturant is removed by common methods of directdilution and one-step *** our experiment,the problem of precipitation was resolved by an improvedgradual dialysis method,which gradually decreased the concentration of ***,fully denaturedRubisco subunits still could not be reassembled into holoenzyme using gradual dialysis unless chaperonin 60was *** restored activity of reassembled Rubisco was approximately 8% of natural *** of reassembled Rubisco increased greatly when heat shock protein 70 was present in the *** and Mg2+ were unnecessary for in vitro reassembly of Rubisco,and Mg2+ inhibited the *** reassembly was weakened when ATP,Mg2+ and K+ existed together in the reassembly process.

关键词： Rubisco reassembly gradual dialysis chaperonin 60 Hsp70

Association between Alcohol, Smoking and HLA-DQA1*0201 Genotype in Psoriasis

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Acta Biochimica et Biophysica Sinica 2004年第9期36卷 597-602页

作者： Guang-Yong ZHENG1,2#, Sheng-Cai WEI3#, Tie-liu shi1, and Yi-Xue li1* 1Bioinformatics Center, shanghai institutes for biological sciences, chinese academy of sciences, Graduate School of the chinese academy of sciences, shanghai 200031, china Research Center for Proteome Analysis, institute of Biochemistry and Cell Biology, shanghai institutes for biological sciences, chinese academy of sciences, Graduate School of the chinese academy of sciences, shanghai 200031, china 3Guangzhou institute of Dermatology, Guangzhou 510095, china Bioinformatics Center Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200031 China Research Center for Proteome Analysis Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200031 China Guangzhou institute of DermatologyGuangzhou 510095 China

Psoriasis is a chronic skin disease triggered by genetic, environment or other risk factors such as infection, drugs, stress, moisture, alcohol, and smoking. A major psoriasis susceptibility locus at 6p21.3 has been identified. Further studies found that HLA-DQA1*0201 allele was associated with psoriasis. However, there were few data exploring an association between the environmental factors and susceptibility genes. In this study, the samples of 189 patients with psoriasis and 333 healthy controls were collected with their consent and were carried on analysis through polymerase chain reaction sequence-specific primer (PCR- SSP) method. The proportion of male psoriasis patients engaging in the smoking and alcohol was much higher than that of the control group (P6.91, P<1.0×10–4).

关键词： psoriasis HLA-DQA1*0201 allele smoking alcohol association

Transcriptional, post-transcriptional and post-translational regulations of gene expression during leaf polarity formation

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Cell Research 2007年第6期17卷 512-519页

作者： lin Xu li YangHai Huang National Laboratory of Plant Molecular Genetics, shanghai institute of Plant physiology and Ecology, shanghai institutes for biological sciences, chinese academy of sciences, 300 fenglin Road, shanghai 200032, china National Laboratory of Plant Molecular Genetics Shanghai Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China Present address: IBMP CNRS Universite Louis Pasteur 67084 Strasbourg France Present address: Department of Biology University of Pennsylvania Philadelphia PA 19104 USA

Leaf morphogenesis requires the establishment of adaxial-abaxial polarity after primordium initiation from the shootapical meristem (SAM). Several families of transcription factors are known to play critical roles in promoting adaxial orabaxial leaf fate. Recently, post-transcriptional gene silencing pathways have been shown to regulate the establishmentof leaf polarity, providing novel and exciting insights into leaf development. For example, microRNAs (miR165/166)and a trans-acting siRNA (TAS3-derived tasiR-ARF) have been shown to repress the expression of several key transcrip-tion factor genes. In addition, yet another level of regulation, post-translational regulation, has been revealed recently bystudies on the role of the 26S proteasome in leaf polarity. Although our understanding regarding the molecular mecha-nisms underlying establishment of adaxial-abaxial polarity has greatly improved, there is still much that remains *** review aims to discuss recent progress, as well as the remaining questions, regarding the molecular mechanismsunderlying leaf polarity formation.

关键词： Arabidopsis leaf development polarity formation

Opioid receptor antagonists increase [Ca2+]i in rat arterial smooth muscle cells in hemorrhagic shock

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Acta Pharmacologica Sinica 2004年第3期25卷 141-146页

作者： li KAI,Zhong-feng WANG2,yu-liang shi2,3,liang-ming liU,De-yao HU institute of Surgery,Department of Pharmacology,Third Military Medical University,Chongqing 400042 Key Laboratory of 2 Neurobiology,shanghai institutes for biological sciences,chinese academy of sciences,shanghai 200031,china institute of Surgery Department of Pharmacology Third Military Medical University 重庆 400042 中科院上海生命科学院 Key Laboratory of Neurobiology CAS 上海 200031

AIM: To examine the effects of opioid receptor antagonists and norepinephrine on intracellular free Ca2+ concentra- tion ([Ca2+]i) in mesenteric arterial (MA) smooth muscle cells (SMC) isolated from normal and hemorrhagic shocked rats in the vascular hyporesponse stage. METHODS: The rat model of hemorrhagic shock was made by withdrawing blood to decrease the artery mean blood pressure to 3.73-4.26 kPa and keeping at the level for 3 h. [Ca2+]i of vascular smooth muscle cells (VSMC) were detected by the laser scan confocal microscopy. RESULTS: In the hyporesponse VMSC of rats in hemorrhagic shock, selective δ-, κ-, and μ-opioid receptor antagonists (naltrindole, nor-binaltorphimine, and β-funaltrexamine, 100 nmol/L) as well as norepinephrine 5 μmol/L significantly increased [Ca2+]i by 47 %±13 %, 37 %±14 %, 33 %±10 %, and 54 %±17 %, respectively, although their effects were lower than those in the normal rat cells (the increased values were 148 %±54 %, 130 %±44 %, 63 %±17 % and 110 %±38 %, respectively); and the norepinephrine-induced increase in [Ca2+]i was further augmented by three δ-, κ-, and μ-opioid receptor antagonists (50 nmol/L, respectively) application (from 52 %±16 % to 99 %±29 %, 146 %±54 % and 137 %±47 %, respectively). CONCLUSION: The disorder of [Ca2+]i regulation induced by hemorrhagic shock was mediated by opioid receptor and α-adrenoceptor, which may be partly responsible for the vascular hyporesponse, and the opioid receptor antagonists improved the response of resistance arteries to vascu- lar stimulants in decompensatory stage of hemorrhagic shock.

关键词： hemorrhagic shock vascular smooth muscle opioid receptors calcium

Comparative Analysis of Two-component Signal Transduction System in Two Streptomycete Genomes

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Acta Biochimica et Biophysica Sinica 2007年第5期39卷 317-325页

作者： WU WEI~(1,3#), Weihua WANG~(2#), Zhiwei CAO~(3#), Hong yu~3, Xiaojing WANG~1, Jing ZHAO~3, Hao TAN~3, Hao XU~3, Weihong JIANG~(2*), and Yixue li~(1,3*) 1 Bioinformation Center, Key Lab of Systems Biology, shanghai institutes for biological sciences, chinese academy of sciences, Graduate School of the chinese academy of sciences, shanghai 200031, china 2 Laboratory of Molecular Microbiology, institute of Plant physiology and Ecology, shanghai institutes for biological sciences, chinese academy of sciences, Graduate School of the chinese academy of sciences, shanghai 200032, china 3 shanghai Center for Bioinformation Technology, shanghai 200235, china Bioinformation Center Key Lab of Systems Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200031 China shanghai Center for Bioinformation Technologyshanghai 200235 China Laboratory of Molecular Microbiology Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200032 China

Species of the genus Streptomyces are major bacteria responsible for producing most natural antibiotics. Streptomyces coelicolor A3(2) and Streptomyces avermitilis were sequenced in 2002 and 2003, respectively. Two-component signal transduction systems (TCSs), consisting of a histidine sensor kinase (SK) and a cognate response regulator (RR), form the most common mechanism of transmembrane signal transduction in prokaryotes. TCSs in S. coelicolor A3(2) have been analyzed in detail. Here, we identify and classify the SK and RR of S. avermitilis and compare the TCSs with those of S. coelicolor A3(2) by computational approaches. Phylogenetic analysis of the cognate SK-RR pairs of the two species indicated that the cognate SK-RR pairs fall into four classes according to the distribution of their orthologs in other organisms. In addition to the cognate SK-RR pairs, some potential partners of non-cognate SK-RR were found, including those of unpaired SK and orphan RR and the cross-talk between different components in either strain. Our study provides new clues for further exploration of the molecular regulation mechanism of streptomycetes with industrial importance.

关键词： Streptomyces two-component system cross-talk phylogenetic analysis

Sequencing and Analysis of a Genomic Fragment Provide an Insight into the Dunaliella viridis Genomic Sequence

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Acta Biochimica et Biophysica Sinica 2006年第11期38卷 812-820页

作者： Xiao-Ming SUN~(1#) yuan-Ping TANG~(1#) Xiang-Zong MENG~1 Wen-Wen ZHANG~1 Shan li~1 Zhi-Rui DENG~1 Zheng-Kai XU~(1,2) Ren-Tao SONG~1* 1 shanghai Key Laboratory of Bio-energy Crops,School of life sciences,shanghai University,shanghai 200444,china 2 institute of Plant physiology & Ecology,shanghai institutes for biological sciences,chinese academy of sciences,shanghai 200032,china shanghai Key Laboratory of Bio-energy Crops School of Life Sciences Shanghai UniversityShanghai 200444 China institute of Plant physiology & Ecology Shanghai Institutes for Biological Sciences Chinese Academy of SciencesShanghai 200032 China

Dunaliella is a genus of wall-less unicellular eukaryotic green *** exceptional resistancesto salt and various other stresses have made it an ideal model for stress tolerance ***,very littleis known about its genome and genomic *** this study,we sequenced and analyzed a 29,268 bpgenomic fragment from DunalieIla *** fragment showed low sequence homology to the *** the nucleotide level,only a segment with significant sequence homology to 18S rRNA *** fragment contained six putative genes,but only one gene showed significant homology at theprotein level to GenBank *** average GC content of this sequence was 51.1%,which was muchlower than that of close related green algae Chlamydomonas （65.7%）.Significant segmental duplicationswere found within this *** duplicated sequences accounted for about 35.7% of the *** amounts of simple sequence repeats （microsatellites） were found,with strong bias towards（AC）_n type （76%）.Analysis of other Dunaliella genomic sequences in the GenBank database （total 25,749bp） was in agreement with these *** sequence features made it difficult to sequence Dunaliellagenomic *** investigation should be made to reveal the biological significance of these uniquesequence features.

关键词： Dunaliella viridis genomic sequence sequence feature sequence duplication simple sequence repeat

p53 Promoter-based Reporter Gene in vitro Assays for Quick Assessment of Agents with Genotoxic Potential

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Acta Biochimica et Biophysica Sinica 2007年第3期39卷 181-186页

作者： Huaixing li~(2#) Ke shi~(1#) Ruiwen CHEN~1 Yan HE~1 Dan WU~1 Shuhan SUN~1* 1 Department of Medical Genetics.Division of Basic Medical sciences,Second Military Medical University,shanghai 200433,china 2 institute for Nutritional sciences.shanghai institutes of biological sciences,chinese academy of sciences,shanghai 200031,china institute for Nutritional sciences Shanghai Institutes of Biological Sciences Chinese Academy of Sciences Shanghai 200031 China Department of Medical Genetics Division of Basic Medical Sciences Second Military Medical University Shanghai 200433 China

The p53 promoter-based green fluorescent protein(GFP)and luciferase reporter gene assayshave been established for detecting DNA damage induced by genotoxic *** evaluate the system,NIH3T3 cells transfected with either pHP53-GFP or pMP53-GFP construct were treated with mitomycin *** of the GFP reporter gene was significantly and specifically induced in the cellsexposed to mitomycin or *** we treated NIH3T3 cells harboring pHP53-Luc or pMP53-Luc vector with mitomycin,5-fluorouracil or cisplatin at various ***,exposure of thecells to these agents with genotoxic potentials resulted in a dose-dependent induction in luciferase reportergene ***,these in vitro reporter gene assays could provide an ideal system for quick assess-ment or screening of agents with genotoxic potential.

关键词： p53 promoter luciferase green fluorescence protein(GFP) genotoxic agent