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Novel cyclophilin D inhibitors derived from quinoxaline exhibit highly inhibitory activity against rat mitochondrial swelling and Ca2+ uptake/release

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Acta Pharmacologica Sinica 2005年第10期26卷 1201-1211页

作者： Hong-xia GUO~(2,3), Feng WANG~(2,3), Kun-qian YU~3, Jing CHEN~3, Dong-lu BAI~3, Kai-xian CHEN~3, Xu SHEN~(3,4,5), Hua-liang JIANG~(3,4,5) ~3 Drug Discovery and Design center, State Key Laboratory of Drug research, shanghai Institute of Materia Medica, shanghai institutes for Biologieal sciences Graduate School of the chinese academy of sciences, chinese academy of sciences, shanghai 201203, china ~4 School of Pharmacy, East china University of Science and Technology shanghai 200237, china Drug Discovery and Design center State Key Laboratory of Drug Research Shanghai Institute of Materia Medica Shanghai Institutes for Biological Sciences Graduate School of the Chinese Academy of Sciences Shanghai 201203 China.

Aim: To investigate methods for identifying specific cyclophilin D （CypD） inhibitors derived from quinoxaline, thus developing possible lead compounds toinhibit mitochondrial permeability transition （MPT） pore opening. Methods: Kinetic analysis of the CypD/inhibitor interaction was quantitatively performed byusing surface plasmon resonance （SPR） and fluorescence titration （FT） ***50values of these inhibitors were determined by PPIase inhibition activity ***: All the equilibrium dissociation constants （KD） of the seven compoundsbinding to CypD were below 10 μmol/L. The IC50values were all consistent withthe SPR and FT results. Compounds GW2, 5, 6, and 7 had high inhibition activitiesagainst Ca2+-dependent rat liver mitochondrial swelling and Ca2+uptake/*** GW5 had binding selectivity for CypD over CypA. Conclusion: Theagreement between the measured IC50values and the results of SPR and FT suggests that these methods are appropriate and powerful methods for fyingCypD inhibitors. The compounds we screened using these methods （GW1-7） arereasonable CypD inhibitors. Its potent ability to inhibit mitochondrial swelling and the binding selectivity of GW5 indicates that GW5 could potentially be used for inhibiting MPT pore opening.

关键词： cyclophilin quinoxalines surface plasmon resonance mitochondrial permeability transition: fluorescence titration inhibitor

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Thansgenic peanut plants obtained by particle bombardment via somatic embryogenesis regeneration system

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Cell research 2001年第2期 156-160页

作者： DENG XIANG YANG, ZHI MING WEI, HAI LONG AN Institute of Plant Physiology and Ecology, shanghai institutes for biological sciences, chinese academy sciences, shanghai 200032, china 中科院上海植物生理生态所上海 200032

After pre-culture and treatment of osmosis, cotyledons of immature peanut (Arachis hypogaea L.) zygotic embryos were transformed via particle bombardment with a plasmid containing a chimeric hph gene conferring resistance to hygromycin and a chimeric intron-gus gene. Selection for hygromycin resistant calluses and somatic embryos was initiated at 10th d post-bombardment on medium containing 10-25 mg/L hygromycin. Under continuous selection, hygromycin resistant plantlets were regenerated from somatic embryos and were recovered from nearly 1.6% of the bombarded cotyledons. The presence and integration of foreign DNA in regenerated hygromycin resistant plants was confirmed by PCR (polymerase chain reaction) for the intron-gus gene and by Southern hybridization of the hph gene. GUS enzyme activity was detected in leaflets from transgenic plants but not from control, non-transformed plants. The production of transgenic plants are mainly based on a newly improved somatic embryogenesis regeneration system developed by us.

关键词： Peanut (Arachis hypogaea) somatic embryo hygromycin resistance particle bombardment.

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Answering a century old riddle: brachydactyly type Al

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Cell research 2004年第3期 179-187页

作者： Bo GAO, Lin HE Institute for Nutritional sciences, shanghai institutes for biological sciences, chinese academy of sciences, shanghai 200031, china. Neuropsychiatric and Human Genetics Group, Bio-X Life Science research center, shanghai Jiaotong University, shanghai 200030, china. Department of Biochemistry, Faculty of Medicine, The University of Hong Kong, Hong Kong, china.

In 1903, Farabee analyzed the heredity of the human digital malformation, brachydactyly, the first recorded disorder of the autosomal dominant Mendelian trait. In 1951, Bell classified this type of brachydactyly as type A1 (BDA1). Over 100 cases from different ethnic groups have so far been reported. However, the real breakthrough in identifying the cause of BDA1 has only taken place in the last few years with the progress of the mapping and identification of one of the genes responsible for this disorder, thus providing an answer for a century old riddle. In this article, we attempt to review the current state of knowledge on the genetic features of BDA1 with its century-old history and signalling pathway of IHH, and also discuss genotype-phenotype correlation not only of BDA1, but also of all types of brachydactyly.

关键词： Brachydactyly IHH GDF5 ROR2 BMPRIB.

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Different expression patterns of duplicated PHANTASTICA-like genes in Lotus japonicus suggest their divergent functions during compound leaf development

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Cell research 2005年第8期15卷 665-677页

作者： Jiang Hong LUO1, Jun YAN1, Lin WENG1, Jun YANG1, Zhong ZHAO3, Jiang Hua CHEN1, Xiao He HU1, Da LUO1,2,* 1National Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, shanghai institutes for Bio- logical sciences, and Graduate School of the chinese academy of sciences, chinese academy of sciences, 300 Fenglin Road, shanghai 200032, china 2School of Life Science and biotechnology, shanghai Jiao Tong University, 1954 Huashan Road, shanghai 200030, china 3Department of Biochemistry, School of Life sciences, Fudan University, shanghai 200433, china National Laboratory of Plant Molecular Genetics Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences and Graduate School of the Chinese Academy of Sciences Chinese Academy of Sciences Shanghai China Department of Biochemistry School of Life Sciences Fudan University Shanghai China School of Life Science and biotechnology Shanghai Jiao Tong University Shanghai China

Recent studies on leaf development demonstrate that the mechanism on the adaxial-abaxial polarity pattern formation could be well conserved among the far-related species, in which PHANTASTICA (PAHN)-like genes play important roles. In this study, we explored the conservation and diversity on functions of PHAN-like genes during the compound leaf development in Lotus japonicus, a papilionoid legume. Two PHAN-like genes in L. japonicus, LjPHANa and LjPHANb, were found to originate from a gene duplication event and displayed different expression patterns during compound leaf development. Two mutants, reduced leaflets1 (rel1) and reduced leaflets3 (rel3), which exhibited decreased adaxial identity of leaflets and reduced leaflet initiation, were identified and investigated. The expression patterns of both LjPHANs in rel mutants were altered and correlated with abnormalities of compound leaves. Our data suggest that LjPHANa and LjPHANb play important but divergent roles in regulating adaxial-abaxial polarity of compound leaves in L. japonicus.

关键词： adaxial identity LjPHANa LjPHANb gene duplication rel mutants Lotus japonicus.

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对水稻 4号染色体上一个编码二氢黄酮醇还原酶类似蛋白基因簇的结构和进化分析(英文)

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生物化学与生物物理学报 2002年第6期34卷 685-689页

作者：雷海燕周波张宇洪国藩韩斌中国科学院上海生命科学研究院国家基因研究中心中国科学院上海生命科学研究院国家基因研究中心上海200233 中国科学院上海生命科学研究院生物化学与细胞生物学研究所上海200031 上海200233

通过对水稻 4号染色体一段 32 3kb的序列测定和分析 ,在其中 56kb的区域内发现了一个由 7个编码二氢黄酮醇还原酶 (DFR)类似蛋白基因组成的基因簇。这 7个基因在基因簇中串联排列 ,每个基因都由 6个外显子和 5个内含子组成。这 7个基因... 详细信息

通过对水稻 4号染色体一段 32 3kb的序列测定和分析 ,在其中 56kb的区域内发现了一个由 7个编码二氢黄酮醇还原酶 (DFR)类似蛋白基因组成的基因簇。这 7个基因在基因簇中串联排列 ,每个基因都由 6个外显子和 5个内含子组成。这 7个基因的预测蛋白质序列都和DFR以及BANYULS蛋白序列类似。DFR和BANYULS都是植物次生代谢类黄酮醇生物合成途径中的结构基因 ,它们的缺失或突变都会造成植物花色素合成代谢的不正常。RT PCR实验证明这 7个基因在水稻的 5个组织中表达不同。文中讨论了这 7个基因的结构和功能特性以及它们的进化关系。

关键词：基因簇水稻拟南芥二氢黄酮醇还原酶(DFR) 水稻类黄酮生物合成酶(OsEFS)

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Skeletal myogenesis by human embryonic stem cells

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Cell research 2006年第8期16卷 713-722页

作者： Jun Ke Zheng~1 Yi Wang~1 Aditi Karandikar~1 Qian Wang~1 Hui Gai~1 Ai Lian Liu~1 Chao Peng~1 Hui Zhen Sheng~(1,2) ~1center for Developmental Biology,Xinhua Hospital,School of Medicine,shanghai Jiao Tong University,1665 Kong Jiang Road,shanghai 200092,china:~2Laboratory of Stem Cell Biology,Institute of Biochemisny and Cell Biology,shanghai institutes of biological sciences,chinese academy of sciences,shanghai 200037,china center for Developmental Biology Xinhua Hospital School of Medicine Shanghai Jiao Tong University Shanghai China Laboratory of Stem Cell Biology Institute of Biochemistry and Cell Biology Shanghai Institutes of Biological Sciences Chinese Academy of Sciences Shanghai China

We have examined the myogenic potential of human embryonic stem(hES)cells in a xeno-transplantation *** we show that precursors differentiated from hES cells can undergo myogenesis in an adult environmentand give rise to a range of cell types in the myogenic *** study provides direct evidences that hES cells canregenerate both muscle and satellite cells in viro and are another promising cell type for treating muscle degenerativedisorders in addition to other myogenic cell types.

关键词： Skeletal myogenesis embryonic stem cell differentiation

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The spike protein of severe acute respiratory syndrome (SARS) is cleaved in virus infected Vero-E6 cells

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Cell research 2004年第5期14卷 400-406页

作者： Xiao Dong WU 1*, Bo SHANG1*, Rui Fu YANG2*, Hao YU3*, Zhi Hai MA1*, Xu SHEN3*, Yong Yong JI1, Ying LIN , Ya Di WU1, Guo Mei LIN1, Lin TIAN1, Xiao Qing GAN1, Sheng YANG2, Wei Hong JIANG2, 1 Er Hei DAI4, Xiao Yi WANG4, Hua Liang JIANG3, You Hua XIE1, Xue Liang ZHU1, Gang PEI1, Lin LI1, Jia Rui WU1**, Bing SUN 1** 1Institute of Biochemistry and Cell Biology, shanghai institutes for biological sciences, chinese academy of sciences, 320 Yueyang Road, shanghai 200031, china. 2Institute of Microbiology and Epidemiology, Institute of Basic Medical sciences, academy of Military Medical sciences, Beijing 100071, china. 3Institute of Plant Physiology and Ecology, shanghai institutes for biological sciences, chinese academy of sciences, 300 Fenglin Road, shanghai 200031, china. 3Institute of Materia Medica, shanghai institutes for biological sciences, chinese academy of sciences, 320 Taiyuan Road, shanghai 200031, china. Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yueyang Road Shanghai 200031 China.

Spike protein is one of the major structural proteins of severe acute respiratory syndrome-coronavirus. It is essential for the interaction of the virons with host cell receptors and subsequent fusion of the viral envelop with host cell membrane to allow infection. Some spike proteins of coronavirus, such as MHV, HCoV-OC43, AIBV and BcoV, are proteolytically cleaved into two subunits, S1 and S2. In contrast, TGV, FIPV and HCoV-229E are not. Many studies have shown that the cleavage of spike protein seriously affects its function. In order to investigate the maturation and proteolytic processing of the S protein of SARS CoV, we generated S1 and S2 subunit specific antibodies (Abs) as well as N, E and 3CL protein-specific Abs. Our results showed that the antibodies could efficiently and specifically bind to their corresponding proteins from *** expressed or lysate of SARS-CoV infected Vero-E6 cells by Western blot analysis. Furthermore, the anti-S1 and S2 Abs were proved to be capable of binding to SARS CoV under electron microscope observation. When S2 Ab was used to perform immune precipitation with lysate of SARS-CoV infected cells, a cleaved S2 fragment was detected with S2-specific mAb by Western blot analysis. The data demonstrated that the cleavage of S protein was observed in the lysate, indicating that proteolytic processing of S protein is present in host cells.

关键词： SARS-CoV spike protein nucleocapsid protein envelope protein 3CL protease polyclonal antibody.

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Gene expression and metabolite profiles of cotton fiber during cell elongation and secondary cell wall synthesis

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Cell research 2007年第5期17卷 422-434页

作者： Jin-Ying Gou1,2,3 Ling-Jian Wang1,2 Shuang-Ping Chen4 Wen-Li Hu1,2 Xiao-Ya Chen1,2 1 National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, shanghai institutes for biological sciences, chinese academy of sciences, 300 Fenglin Road, shanghai 200032, china 2 CAS center of shanghai Chenshan Botanical Garden, 300 Fenglin Road, shanghai 200032, china 3 Graduate School of chinese academy of sciences, shanghai 200032, china 4 Department of Computer Science and Technology, University of Science and Technology of china, Hefei 230026, china National Key Laboratory of Plant Molecular Genetics Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China CAS center of shanghai Chenshan Botanical Garden Shanghai China Graduate School of chinese academy of sciences Shanghai China Department of Computer Science and Technology University of Science and Technology of China Hefei China

Cotton fibers elongate rapidly after initiation of elongation, eventually leading to the deposit of a large amount ofcellulose. To reveal features of cotton fiber cells at the fast elongation and the secondary cell wall synthesis stages, wecompared the respective transcriptomes and metabolite profiles. Comparative analysis of transcriptomes by cDNA arrayidentified 633 genes that were differentially regulated during fiber development. Principal component analysis (PCA)using expressed genes as variables divided fiber samples into four groups, which are diagnostic of developmental *** grouping results are also found if we use non-polar or polar metabolites as variables for PCA of developingfibers. Auxin signaling, wall-loosening and lipid metabolism are highly active during fiber elongation, whereas cellulosebiosynthesis is predominant and many other metabolic pathways are downregulated at the secondary cell wall synthesisstage. Transcript and metabolite profiles and enzyme activities are consistent in demonstrating a specialization process ofcotton fiber development toward cellulose synthesis. These data demonstrate that cotton fiber cell at a certain stage has itsown unique feature, and developmental stages of cotton fiber cells can be distinguished by their transcript and metaboliteprofiles. During the secondary cell wall synthesis stage, metabolic pathways are streamed into cellulose synthesis.

关键词： cotton fiber transcriptome metabolite profile auxin cell elongation cellulose synthesis

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Nuclear reprogramming: the strategy used in normal development is also used in somatic cell nuclear transfer and parthenogenesis

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Cell research 2007年第2期17卷 135-150页

作者： Tianlong Gao~(1,*) Junke Zheng~(2,*) Fengying Xing~(2,*) Haiyan Fang~2 Feng Sun~1 Ayong Yan~2 Xun Gong~2 Hui Ding~3 Fan Tang~2 Hui Z Sheng~2 ~1Program for Graduation Studies,shanghai institutes for biological sciences,chinese academy of sciences,shanghai 200031,china ~2center for Developmental Biology,Xinhua Hospital,shanghai Jiao Tong University,School of Medicine,1665 Kong Jiang Road,shanghai 200092,china ~3School of Animal Science and Technology,Yangzhou University,Yangzhou 225009,china Program for Graduation Studies Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China center for Developmental Biology Xinhua Hospital Shanghai Jiao Tong University School of Medicine Shanghai China School of Animal Science and Technology Yangzhou University Yangzhou China

Somatic cell nuclear transfer(SCNT)and parthenogenesis are alternative forms of reproduction and development,building new life cycles on differentiated somatic cell nuclei and duplicated maternal chromatin,*** thepreceding paper(Sun F,et al.,Cell Res 2007;17:117-134.),we showed that an"erase-and-rebuild"strategy is used innormal development to transform the maternal gene expression profile to a zygotic ***,we investigate if the samestrategy also applies to SCNT and *** relationship between chromatin and chromatin factors(CFs)during SCNT and parthenogenesis was examined using immunochemical and GFP-fusion protein *** fromthese studies indicated that soon after nuclear transfer,a majority of CFs dissociated from somatic nuclei and were re-distributed to the cytoplasm of the *** erasure process in oogenesis is recaptured during the initial phase in *** CFs entered pseudo-pronuclei shortly after their *** parthenogenesis,all parthenogenotes underwentnormal oogenesis,and thus had removed most CFs from chromosomes before the initiation of *** CFswere subsequently re-associated with female pronuclei in time and sequence similar to that in fertilized *** these data,we conclude that the"erase-and-rebuild"process observed in normal development also occurs in SCNTand in parthenogenesis,albeit in altered *** process is responsible for transcription reprogramming in ***"erase"process in SCNT is compressed and the efficiency is compromised,which likely contribute tothe developmental defects often observed in nuclear transfer(nt)***,results from this study indicatedthat the cytoplasm of an egg contains most,if not all,essential components for assembling the zygotic program and canassemble them onto appropriate diploid chromatin of distinct origins.

关键词： nuclear reprogramming SCNT parthenogenesis chromatin factors chromatin embryogenesis

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Embryonic stem cells generated by nuclear transfer of human somatic nuclei into rabbit oocytes

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Cell research 2003年第4期13卷 251-263页

作者： YING CHEN, ZHI Xu HE, AILIAN LIU, KAI WANG, WEN WEI MAO, JIAN XIN CHU, YONG LU, ZHENG Fu FANG, YING TANG SHI, QING ZHANG YANG, DA YUAN CHEN, MIN RANG WANG, JIN SONG LI, SHAO LIANG HUANG, XIANG YIN KONG, YAO ZHOU SHI, Zm QIANG WANG, JIA HuI XIA, ZHI GAO LONG, ZHI GANG XUE, WEN XIANG DING, HuI ZHEN SHENG center for Developmental Biology, shanghai Second Medical University, shanghai 200092, china Laboratory of Stem Cell Biology, Institute of Biochemistry and Cell Biology, shanghai institutes for biological sciences, chinese academy of sciences, 1665 Kong Jiang Road, shanghai 200092, china center for Stem Cell research, the Second Affiliated Hospital, Sun Yat-Sen University, Guangzhou 510120,china State Key Laboratory of Reproductive Biology, Institute of Zoology, chinese academy of sciences, 19 Zhong-guan Cun Road, Haidian District, Beijing 100080, china Health Science center, shanghai institutes of biological sciences, chinese academy of sciences and shanghai Second Medical University, shanghai 200025, china National Laboratory of Medical Genetics, Central South University, 88 Xiangya Road, Changsha 410078, china Xinhua Hospital, shanghai Second Medical University, 1665 Kong Jiang Road, shanghai 200092, china center for Developmental Biology Shanghai Second Medical University Shanghai 200092 China Laboratory of Stem Cell Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 1665 Kong Jiang Road Shanghai 200092 China center for Stem Cell research the Second Affiliated Hospital Sun Yat-Sen University Guangzhou 510120 China State Key Laboratory of Reproductive Biology Institute of Zoology Chinese Academy of Sciences 19 Zhong-guan Cun Road Haidian District Beijing 100080 China Health Science center Shanghai Institutes of Biological Sciences Chinese Academy of Sciences and Shanghai Second Medical University Shanghai 200025 China National Laboratory of Medical Genetics Central South University 88 Xiangya Road Changsha 410078 China Xinhua Hospital Shanghai Second Medical University 1665 Kong Jiang Road Shanghai 200092 China

To solve the problem of immune incompatibility, nuclear transplantation has been envisaged as a means to produce cells or tissues for human autologous transplantation. Here we have derived embryonic stem cells by the transfer of human somatic nuclei into rabbit oocytes. The number of blastocysts that developed from the fused nuclear transfer was comparable among nuclear donors at ages of 5, 42, 52 and 60 years, and nuclear transfer (NT) embryonic stem cells (ntES cells) were subsequently derived from each of the four age groups. These results suggest that human somatic nuclei can form ntES cells independent of the age of the donor. The derived ntES cells are human based on karyotype, isogenicity, in situ hybridization, PCR and immunocytochemistry with probes that distinguish between the various species. The ntES cells maintain the capability of sustained growth in an undifferentiated state, and form embryoid bodies, which, on further induction, give rise to cell types such as neuron and muscle, as well

关键词： nuclear transfer (NT) somatic cell nuclear transfer (SCNT) embryonic stem cells (ES cell) therapeutic cloning rabbit oocyte.

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