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Effects of genistein, daidzein and glycitein on the osteogenic and adipogenic differentiation of bone marrow stromal cells and on the adipogenic trans-differentiation of osteoblasts

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Progress in Natural science:Materials International 2005年第11期15卷 993-999页

作者： LI Xianghui, ZHANG Jinchao, SUI Senfang and YANG Mengsu (State key laboratory of {2., department of biological sciences and biotechnology, tsinghua university, beijing 100084, china department of Chemistry, College of Chemistry & Environmental science, Hebei {12., Baoding 071002. china department of Biology and Chemistry, City {12. of Hong Kong, Hong Kong, china) State key laboratory of {2. Department of Biological Sciences and Biotechnology Tsinghua University Beijing 100084 China department of Chemistry College of Chemistry and Environmental Science Hebei University Baoding 071002 China department of Biology and Chemistry City University of Hong Kong Hong Kong Hong Kong

The 3-(4,5-dimethylthiazol-2.yl)-2.5-diphenyl tetrazolium bromide (MTT), alkaline phosphatase (ALP) activity and oil red O assays were used to examine the effects of genistein, daidzein and glycitein on the osteogenic and adipogenic differentiation of primary mouse bone marrow stromal cells (MSCs) and the adipogenic trans-differentiation of primary mouse osteoblasts. The results indicated that daidzein, genistein and glycitein at concentrations from 1×10-8 mol/L to 1×10-5mol/L promoted the proliferation of MSCs and osteoblasts; genistein, daidzein and glycitein promoted osteogenic differentiation and inhibited adipogenic differentiation of MSCs, and inhibited adipocytic transdifferentiation of osteoblasts at appropriate concentrations as 17β-estradiol. It suggests that genistein, daidzein and glycitein regulate a dual differentiational process of MSCs into the osteogenic and adipogenic lineages, and trans-differentiational process of primary osteoblasts into the adipocyte lineages, causing a lineage shift toward osteoblast. Protective effects of them on bone may be mediated by a reversal of adipogenesis which may promote the proliferation, differentiation and mineralization of osteoblasts, and make adipocytes secrete less cytokines which may promote osteoclast formation and activation. In addition, the results also indicated that genistein, daidzein and glycitein may be helpful in preventing the development of steroid induced osteonecrosis.

关键词： genistein daidzein glycitein bone marrow stromal cells osteoblasts osteogenic differentiation adipogenic differentiation adipogenic trans-differentiation.

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Two forms of the membrane-bound state of the first C2.domain (C2.) of synaptotagminⅠand calcium-triggered membrane insertion

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Chinese science Bulletin 2003年第17期48卷 1822-1827页

作者： HE Yuhong, LI Xianghui, WANG Fu, XUE Yi & SUI Senfang department of biological {2.s & biotechnology, State key Labora-tory of biomembrane, tsinghua university, beijing 100084, china department of biological {2.s and biotechnology State Key Laboratory of Biomembrane Tsinghua University Beijing 100084 China

The synaptic vesicle protein synaptotagminⅠ (sytⅠ) is a vesicle transmembrane protein present in synap-tic vesicles, which has been proposed as the Ca2. sensor that regulates secretion. The C2. domain is the membrane proximal part of its cytoplasmic domain. The interaction between C2. and lipid bilayer has been considered to be essential for triggering neurotransmitter release. In the pre-sent work, the measurements of membrane surface tension and surface concentration showed that the C2. domain of sytⅠexhibited two membrane-bound states: the surface adsorption state and the membrane insertion state. The sur-face absorption state formed in a Ca2.-independent manner with lower affinity, while the membrane insertion state formed with high affinity was only found in the presence of Ca2.. Both the Ca2.-independent and Ca2.-dependent sytⅠ- membrane interactions required anionic phospholipids, such as phosphatidylserine (PS). When expressed into rat pheo-chromocytoma (PC12. cells and human embryonic kidney (HEK-2.3) cells, as demonstrated by immunofluorescence staining and subcellular fractionation, most of the C2. was found at the plasma membrane, even when the cells were depleted of Ca2. by incubation with EGTA. These results suggested a new molecular mechanism of sytⅠas a Ca2. sensor in membrane fusion. Ca2.-independent surface ad-sorption might attach sytⅠto the release site during the docking or priming step. When intracellular Ca2. increased, sytⅠtriggered the neurotransmitter release following the Ca2.-dependent penetration into the target membrane.

关键词：突触小泡蛋白质膜贯通蛋白质钙离子薄膜插入 C2. 薄膜边界状态

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Zinc up-regulated the expression of the rice metallonthionein gene family and enhanced the zinc tolerance of yeast cells

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Progress in Natural science:Materials International 2007年第10期17卷 1166-1171页

作者： Xu Yufeng, Zhou Gongke, Li Yiqin and Liu Jinyuan(laboratory of Molecular Biology and Protein science laboratory of the Ministry of Education, department of biological sciences and biotechnology, tsinghua university, beijing 100084, china) laboratory of Molecular Biology and Protein science laboratory Department of Biological Sciences and Biotechnology Tsinghua University Beijing 100084 China

Northern blot and functional complementation assay were employed to analyze the effects of zinc on expression of ten rice metallothionein genes (OsMT-Is) in rice seedlings and the growth of yeast cells transformed with OsMT-Is. Northern blot revealed that in shoots of the rice seedlings treated with different Zn2.concentrations, expression of most members of OsMT-I family was increased, except the type 4 OsMT-Is (OsMT-I-4a, 4b and 4c). In roots, Zn2.significantly increased the transcription of OsMT-I-1b and OsMT-I-2., but reduced the trascription of OsMT-I-1a and OsMT-I-3a. When these ten cDNAs were heterologously expressed in zinc sensitive yeast mutant, all transgenic yeasts showed increased tolerance to Zn2., and zinc accumulation in these yeast cells also increased. These indicated that OsMT-I family members might respond to extra Zn2., and they could enhance Zn2. tolerance of cells by direct binding Zn2..

关键词： rice metallothionein induced expression Zn2. tolerance.

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Identification of two distinct transactivation domains in the pluripotency sustaining factor nanog

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Cell Research 2003年第6期13卷 499-502页

作者： GUANG Jin PAN, DUAN QING PEI Institute of Pharmacology, department of biological {2.s and biotechnology and State key Labora tory of biomembrane and membrane biotechnology, tsinghua Institutes of Biomedicine, tsinghua university, 100084 beijing,{12.. E-mail: duanqing@aol.comdepartment of Pharmacology, university of Minnesota, Minneapolis, MN55455, USA department of biological {2.s and biotechnology and State key laboratory of biomembrane and membrane biotechnology Institute of Pharmacology Tsinghua Institutes of Biomedicine Tsinghua University Beijing China department of Pharmacology University of Minnesota Minneapolis USA

Nanog is a newly identified homeodomain gene that functions to sustain the pluripotency of embryonic stem cells. However, the molecular mechanism through which nanog regulates stem cell pluripotency remains unknown. Mouse nanog encodes a polypeptide of 305 residues with a divergent homeodomain similar to those in the NK-2.family. The rest of nanog contains no apparent homology to any known proteins characterized so far. It is hypothesized that nanog encodes a transcription factor that regulates stem cell pluripotency by switching on or off target genes. To test this hypothesis, we constructed fusion proteins between nanog and DNA binding domains of the yeast transcription factor Gal4 and tested the transactivation potentials of these constructs. Our data demonstrate that both regions N- and C- terminal to the homeodomain have transcription activities. Despite the fact that it contains no apparent transactivation motifs, the C-terminal domain is about 7 times as active as the N-terminal one. This unique

关键词： nanog pluripotency stem cells self renewal transactivation domain homeoprotein homeodomain

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Expression profiles and transactivation analysis of a novel ethylene-responsive transcription factor gene GhERF5 from cotton

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Progress in Natural science:Materials International 2009年第5期19卷 563-572页

作者： Longguo Jin, Bo Huang, Hui Li, Jinyuan Liu laboratory of Molecular Biology and MOE laboratory of Protein {2., department of biological {2.s and biotechnology, tsinghua university, beijing 100084, china laboratory of Molecular Biology and MOE laboratory of Protein {2. Department of Biological Sciences and Biotechnology Tsinghua University Beijing 100084 China

Plant AP2.EREBP transcription factors play important roles in plant development and in plant responses to biotic and abiotic stresses. A novel gene for ethylene-responsive element binding protein (EREBP), designated GhERF5, which encodes a protein of 2.5 amino acids, was isolated by RACE-PCR from cotton (Gossypium hirsutum) seedlings. Sequence alignment revealed that GhERF5 contains a typical AP2.ERF domain, and belongs to the B3 subgroup of the ERF subfamily. Particle bombardment assay showed that GhERF5 functions as an in vivo transcription activator in tobacco cells, and it is located in the nuclei of onion epidermis cells. Semi-quantitative RT-PCR revealed that the expression of GhERF5 was highly and rapidly induced when plants were treated with exogenous ethylene, abscisic acid (ABA), salt, cold and drought. Promoter analysis indicated that there are conserved cis-acting elements induced by these stresses in the 50-upstream region of the GhERF5 gene. These results suggest that the GhERF5 gene might play an important role in cotton response to ethylene, ABA and environmental stresses.

关键词： Ethylene-responsive element binding factor (ERF) Abiotic stresses Nuclear localization Transactivation Gossypium hirsutum

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

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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 State key laboratory of biomembrane and membrane biotechnology, tsinghua university, beijing 100084,{2. Medical Systems Biology Research Center Tsinghua University School of Medicine Beijing 100084 China department of biological {2.s and biotechnology Tsinghua University Beijing 100084 China National Engineering Research Center for beijing Biochip Technology 18 Life Science Parkway Beijing 102206 China State key laboratory of {2. 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 {2.s and biotechnology, State key laboratory of biomembrane and membrane biotechnology, beijing 100084, china Institute of Molecular and Cellular Bioscie department of biological {2.s and biotechnology 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, {2. 100093, china 2. department of biological sciences and biotechnology, State key laboratory of biomembrane and membrane biotechnology, tsinghua university, beijing 100084, chinaCorrespondence should be addressed to Kuang Tingyun laboratory of Photosynthesis Basic Research Institute of Botany Chinese Academy of Sciences Beijing China department of biological {2.s and biotechnology 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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Detecting robust gene signature through integrated analysis of multiple types of high-throughput data in liver cancer

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Acta Pharmacologica Sinica 2007年第12期28卷 2005-2010页

作者： Xin-yu ZHANG~(2.3,4,6),Tian-tian LI~(5,6),Xiang-jun LIU~(2.3,4,7)~2.department of biological {2. and biotechnology,~3 School of Biomedicine and ~4 Ministry of Education key laboratory of Bioinformatics,tsinghua university beijing 100084,china ~5 laboratory of Medical Genetics,department of Biology,Harbin Medical university,Harbin150086.china department of biological {2. and biotechnology Tsinghua University Beijing China School of Biomedicine Tsinghua University Beijing China Ministry of Education key laboratory of Bioinformatics Tsinghua University Beijing China department of Biology Laboratory of Medical Genetics Harbin Medical University Harbin China

Aim:To investigate the robust gene signature in liver cancer,we applied anintegrated approach to perform a joint analysis of a highly diverse collection ofliver cancer genome-wide datasets,including genomic alterations and transcrip-tion ***:1-class Significance Analysis of Microarrays coupled withranking score method were used to identify the robust gene signature in livertumor ***:In total,1 62. 051 gene expression measurements from 16public microarrays,2.pairs of serial analyses of gene expression experiments,and2.2.loss of heterozygosity reports obtained from 568 publications were used inthis integrated *** resulting robust gene signatures included 90 genes,which may be of great importance to liver cancer research.A system assessmentanalysis revealed that our integrative method had an accuracy of 92. and acorrelation coefficient value of ***:The system assessment resultsindicated that our method had the ability of integrating the datasets from varioustypes of sources,and eliciting more accurate results,as can be very useful in thestudy of liver cancer.

关键词： liver cancer integrated analysis robust gene signature gene expression map microarray SAGE

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Effect of daidzin,genistin,and glycitin on osteogenic and adipogenic differentiation of bone marrow stromal cells and adipocytic transdifferentiation of osteoblasts

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Acta Pharmacologica Sinica 2005年第9期26卷 1081-1086页

作者： Xiang-hui LI~1 Jin-chao ZHANG~(2.3) Sen-fang SUI~(1,4) Meng-su YANG~(3,4)1 State key laboratory of {2. department of biological sciences and biotechnology,tsinghua university,beijing 100084,china 2.department of Chemistry College of Chemistry and Environmental science,Hebei university,Baoding 071002.china 3 department of Biology and Chemistry,City university of Hong Kong,Hong Kong,china department of biological {2.s and biotechnology State Key Laboratory of Biomembrane Tsinghua University Beijing China department of Chemistry College of Chemistry and Environmental Science Hebei University Baoding China department of Biology and Chemistry City University of Hong Kong Hong Kong China

Aim:To examine the effect of daidzin,genistin,and glycitin on the osteogenic andadipogenic differentiation of bone marrow stromal cells（MSC）and the adipogenictransdifferentiation of ***:MTT test,alkaline phosphatase（ALP）activity measurement,Oil Red O stain and measurement were ***:Daidzin,genistin,and glycitin 1×10-8,5×10-7,1×10-6,5×10-6,and 1×10-5mol/L all promoted the proliferation of primary mouse bone MSC and *** 5×10-7mol/L and genistin 1×10-6mol/L promoted the osteogenesis *** 1×10-8,5×10-7,1×10-6,5×10-6,and 1×10-5mol/L and glycitin 1×10-8,1×10-6,and 1×10-5mol/L inhibited the adipogenesis of ***,genistin,and glycitin 1×10-8,5×10-7,1×10-6,5×10-6,and 1×10-5mol/L all inhibited the adipocytictransdifferentiation of ***:Daidzin,genistin,and glycitinmay modulate differentiation of MSC to cause a lineage shift toward the osteo-blast and away from the adipocytes,and could inhibit adipocytic transdifferen-tiation of *** could also be helpful in preventing the developmentof osteonecrosis.

关键词： daidzin genistin glycitin bone marrow stro-real cells osteogenic differentiation adipogenic differentiation osteoblasts adipocytic transdifferentiation

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