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Cytotoxicity, apoptosis induction, and mitotic arrest by a novel podophyllotoxin glucoside, 4DPG, in tumor cells

Acta Pharmacologica Sinica 2005年第8期26卷 1000-1008页

作者： Yi-lin QI~(2,4), Fan LIAO~(2,4), Chang-qi ZHAO~2, Yong-da LIN~2, Ming-xue ZUO~(2,3,5) ~2 College of Life Sciences, beijing Normal University, beijing 100875 china ~3.key laboratory for molecular biology and gene engineering drugs of beijing, beijing 100875, china College of Life Sciences Beijing Normal University Beijing China key laboratory for molecular biology and gene engineering drugs of beijing Beijing China

Aim: To define the in vitro cytotoxic activities of 4-demethyl-picropodophyllotoxin 7'-O-β-D-glucopyranoside （4DPG）, a new podophyllotoxin glucoside. Methods: Antiproliferation activity was measured in several tumor cell lines by using the microculture tetrazolium MTT assays. Cell cycle distribution was analyzed using flow cytometry and mitosis index assays. Furthermore, transmission electron microscopy, TUNEL, DNA agarose electrophoresis, and activated caspase-3 were used to analyze the induction of apoptotic cell death. Moreover, intracellular changes in the cytoskeleton were detected using immunocytochemistry. Results: 4DPG effectively inhibited the proliferation of cancer cells （HeLa, CNE, SH-SY5Y, and K562 cell lines）. For the K562 cell line, the antiproliferation effect of 4DPG was much more potent than that of etoposide (IC50value: 7.79x10（-9） mol/L for 4DPG vs 2.23x 10（-5） mol/L for etoposide). Further, 4DPG blocked the cell cycle in the mitotic phase. The induction of apoptosis and elevated levels of activated caspase-3 were confirmed in cells treated with 4DPG. The microtubule skeleton of HeLa cells was disrupted immediately after treatment with 4DPG. Conclusion: The cytotoxicity of 4DPG is due to its inhibition of the microtubule assembly of cancer cells at a low concentration, thus inducing apoptosis. These properties qualify 4DPG to be a potential antitumor drug.

关键词： 4-demethyl-picropodophyllotoxin 7’-O-β-D-glucopyranoside podophyllotoxin flow cytometry microtubulin apoptosis

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Metal-amyloid-β peptide interactions: a preliminary investigation of molecular mechanisms for Alzheimer’s disease

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Science china Chemistry 2007年第4期50卷 453-467页

作者： JIAO Yong & YANG Pin key laboratory of Chemical biology and molecular engineering of Ministry of Education, Institute of molecular Science, Shanxi University, Taiyuan 030006, china key laboratory of Chemical biology and molecular engineering of Ministry of Education Institute of Molecular Science Shanxi University Taiyuan China

Although humans have spent exactly 100 years combating Alzheimer’s disease (AD), the molecular mechanisms of AD remain unclear. Owing to the rapid growth of the oldest age groups of the popula-tion and the continuous increase of the incidence of AD, it has become one of the crucial problems to modern sciences. It would be impossible to prevent or reverse AD at the root without elucidating its molecular mechanisms. From the point of view of metal-amyloid-β peptide (Aβ) interactions, we review the molecular mechanisms of AD, mainly including Cu2+ and Zn2+ inducing the aggregation of Aβ, cata-lysing the production of active oxygen species from Aβ, as well as interacting with the ion-channel-like structures of Aβ. Moreover, the development of therapeutic drugs on the basis of metal-Aβ interactions is also briefly introduced. With the increasingly rapid progress of the molecular mechanisms of AD, we are now entering a new dawn that promises the delivery of revolutionary developments for the control of dementias.

关键词： Alzheimer’s disease (AD) amyloid-β peptide Cu2+ Zn2+ molecular mechanisms aggregation active oxygen species ion channel

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Transgenic expression of DwMYB2 impairs iron transport from root to shoot in Arabidopsis thaliana

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Cell Research 2006年第10期16卷 830-840页

作者： Yan-Hong Chen~(1,2,3) Xue-Min Wu~5 Hong-Qing Ling~4 Wei-Cai Yang~1 ~1The key laboratory of molecular and Developmental biology,Institute of genetics and Developmental biology,Chinese Academy of Sciences,beijing 100101 china ~2Graduate University of the Chinese Academy of Sciences,19 Yuquan Road,beijing 100049,china ~3School of Life Science,Nantong University,Nantong 226007 china ~4The State key laboratory of Plant Cell and Chromosome engineering,Institute of genetics and Developmental biology,Chinese Academy of Sciences,beijing 100101,china The key laboratory of molecular and Developmental biology Institute of Genetics and Developmental Biology Chinese Academy of Sciences Beijing China Graduate University of the Chinese Academy of Sciences Beijing China School of Life Science Nantong University Nantong China Department of Botany University of British Columbia 6270 University Blvd. Vancouver Canada The State key laboratory of Plant Cell and Chromosome engineering Institute of Genetics and Developmental Biology Chinese Academy of Sciences Beijing China

In plant,iron uptake and homeostasis are tightly regulated to ensure its absorption from soil and to avoid excessiron in the *** genes involved in this process have been identified during past several years,but there are manyproblems remain unsolved in the genetic regulation of whole plant iron trafficking and *** transcriptionfactors contain tandem repeats of a～50 amino acid DNA-binding motif (R) and are involved in the regulation of manyaspects of plant development,hormone signaling and ***,we report that the ectopic expression of orchidR2R3-MYB gene DwMYB2 in Arabidopsis thaliana confers the transgenic plants hypersensitivity to iron *** DwMYB2 transgenic plants,the iron content in root is two-fold higher compared to that in wild-type root,while thereverse is true in *** imbalance of iron content in root and shoot suggested that the translocation of iron fromroot to shoot was affected by the expression of DwMYB2 in the transgenic ***,gene chip and reversetranscription-polymerase chain reaction analysis revealed that the ferric-chelate reductase gene,AtFRO2,and the irontransporter gene,AtIRT1 and AtIRT2,are up-regulated by DwMYB2 expression,while other potential iron transporterssuch as AtIREG1,AtFRD3 and NRAMP1 are *** addition,the expression of several putative peptidetransporters and transcription factors are also altered in the 35S::DwMYB2 transgenic *** data provide us insightinto the whole plant translocation of iron and identify candidate genes for iron homeostasis in plants despite the fact thata heterologous gene was expressed.

关键词： DwMYB2 iron uptake and homeostasis iron transporters Arabidopsis

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哺乳动物中Kelch超家族蛋白的结构与功能

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生物化学与生物物理进展 2011年第3期38卷 210-217页

作者：肖枫宋宏涛魏群北京师范大学生命科学学院北京市基因工程药物与生物技术重点实验室北京100875

在不同生物体,如病毒、植物、真菌和哺乳动物中,几百种含Kelch构象的蛋白质已经被鉴定出来了,但大部分此类蛋白质的功能未知.而在哺乳动物中,已经有接近41种Kelch蛋白被报道.这些蛋白质在体内参与了很多重要的生理过程.根据Kelch蛋白的... 详细信息

在不同生物体,如病毒、植物、真菌和哺乳动物中,几百种含Kelch构象的蛋白质已经被鉴定出来了,但大部分此类蛋白质的功能未知.而在哺乳动物中,已经有接近41种Kelch蛋白被报道.这些蛋白质在体内参与了很多重要的生理过程.根据Kelch蛋白的域结构不同,把这些哺乳动物中的Kelch蛋白分成了三类(BTB/Kelch蛋白,只含有Kelch结构域的蛋白质,及含其他结构域的Kelch蛋白),并把其已知的功能大致划分为5类:介导蛋白质-蛋白质的相互作用、参与蛋白质降解、信号传导、胞外功能及其他等.另外还认为Keap 1的Kelch结构域其晶体结构应该是研究哺乳动物Kelch蛋白的一个很好的参考模型.

关键词： Kelch蛋白哺乳动物细胞形变蛋白质降解信号传导

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Overexpression of p27^(KIP1)induced cell cycle arrest in G_1 phase and subsequent apoptosis in HCC-9204 cell line

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World Journal of Gastroenterology 2000年第4期6卷 513-521页

作者： Jiang Li Xin Ke Yang Xin Xin Yu Meng Liang Ge Wen Liang Wang Jie Zhang Yun De Hou Department of Pathology,Fourth Military Medical University,Xi’an 710033,Shaanxi Province,china State key laboratory for molecular Virology and genetic engineering,beijing 100052,china Department of Dermatology,beijing Hospital,beijing 100016,china Institute of Radiation Medicine,beijing 100085,china 第四军医大学陕西西安中国预防医学科学院病毒所北京中国预防医学科学院病毒所分子病毒基因工程国家重点实验室北京北京医院北京解放军军事医学科学院放射医学所北京

AIM We have previously reported that inducible over-expresaion of Bak may prolong cell cycle in G1 phase and lead to apoptosis in HCC-9204 cells. This study is to investigate whether p27KIP1 plays an important role in this process. MEHODS In order to elucidate the exact function of p27KIP1 in this process, a zinc inducible p27KIP1 stable transfectant and transient p27KIP1- GFP fusion transfectant were constructed. The effects of inducible p27KIP1 on cell growth, cell cycle arrest and apoptosis were examined in the mock, control pMD vector, and pMD-KIP1 transfected HCC-9204 cells. RESULTS This p27KIP1-GFP transfectant may transiently express the fusion gene. The cell growth was reduced by 35% at 48 h of p27KIP1 induction with zinc treatment as determined by trypan blue exclusion assay. These differences remained the same after 72 h of p27KIP1 expression, p27KIP1 caused cell cycle arrest after 24 h of induction, with 40% increase in G1 population. Prolonged p27KIP1 expression in this cell line induced apoptotic cell death reflected by TUNEL assay. Fourty-eight h and 72 h of p27KIP1 expression showed a characteristic DNA ladder on agarose gel electrophoresis.

关键词： p27^(KIP1) apoptosis cell cycle inducible expression system carcinoma hepatocellular liver neoplasms

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Expressed sequence tag analysis and cloning of trehalose-6-phosphate synthase gene from marine alga Laminaria japonica(Phaeophyta)

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Acta Oceanologica Sinica 2012年第6期31卷 139-148页

作者： XUAN Jinsong 1,4,FENG Yanbin 1,WENG Manli 1,ZHAO Ge 1,SHI Jinfeng 1,YAO Jianting 2,WANG Xiuliang 2,GUO Baotai 3,QIAO Lixian 3,DUAN Delin 2,WANG Bin 1 1 The State key laboratory of Plant Genomics,Institute of genetics and Developmental biology,Chinese Academy of Sciences,beijing 100101,china 2 The State key laboratory of Marine biology,Institute of Oceanology,Chinese Academy of Sciences,Qingdao 266071,china 3 College of Life Sciences,Qingdao Agricultural University,Qingdao 266109,china 4 Department of Biological Science and engineering,School of Chemical and Biological engineering,University of Science and Technology beijing,beijing 100083,china The State key laboratory of Plant Genomics Institute of Genetics and Developmental Biology Chinese Academy of Sciences Beijing China Department of Biological Science and engineering School of Chemical and Biological Engineering University of Science and Technology Beijing Beijing China The State key laboratory of Marine biology Institute of Oceanology Chinese Academy of Sciences Qingdao China College of Life Sciences Qingdao Agricultural University Qingdao China

A high quality cDNA library was constructed from the brown alga Laminaria japonica,with the titer of 1.2×10 5 pfu/*** average insert size of the cDNA library is about 1.6 *** the cDNA library,591 cDNA clones were randomly selected and *** a result,574 EST(expressed sequence tag) sequences were *** of the 574 ESTs were submitted to the dbEST database section of GenBank with the accession numbers from CX942625 to *** cDNA library was screened with a α-32 p labeled 453 bp T P S gene probe,which is a partial sequence yielded from Porphyra *** positive cDNA clones were screened and the sequencing data showed that these four cDNA clones covered majority of *** TPS cDNA *** PCR amplification,sequencing and assembling,the entire ORF(open reading frame) sequence of the T P S gene was obtained,which was named *** encodes a protein containing 908 amino acids with a calculated molecular mass of 101 674 *** LjTPS gene was successfully expressed in *** and *** LjTPS gene has potential application both in plant breeding to stress tolerance and in deciphering the T P S gene function and mechanism to stress tolerance.

关键词： cDNA library comparative study expressed sequence tag(EST) analysis Laminaria japonica trehalose-6-phosphate synthase(TPS) T P S gene

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generation of a recombinant herpes simplex virus which can provide packaging function for recombinant adeno-associated virus

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Chinese Science Bulletin 1999年第8期44卷 715-719页

作者： WU Zhijian, WU Xiaobing and HOU YundeState key laboratory for molecular Virology and genetic engineering , Institute of Virology , Chinese Academy of Preventive Medicine , beijing 100052, china Corresponding author State key laboratory for molecular Virology and genetic engineering Institute of Virology Chinese Academy of Preventive Medicine Beijing China

The generation of a recombinant HSV (rHSV) that can provide packaging function for rAAV production is described. A set of cosmids including cos48, cos28, cos6, cos14 and cos56, which represents the HSV-1 genome was used for generation of this rHSV. Rep and cap genes of AAV-2 were inserted into Xba Ⅰ site of UL2 gene on cos6, generating cos6-rcΔUL2. After being digested with Pac Ⅰ , cos6-rcΔUL2 and the other 4 cosmids were cotrans-fected into BHK-21 cells. The recombinant virus HSV1-rc/ΔUL2 carrying rep and cap genes was generated due to the homologous recombination of the 5 cosmids. The results showed that the existence of rep and cap genes on this rHSV was stable from passage to passage and the rHSV could support the packaging of rAAV either in cells transiently transfected with AAV vector or in stable cell line harboring AAV vector. Further modification of this rHSV and optimization of conditions involved in rAAV preparation may lead to a large-scale production of rAAV in the near future.

关键词： recombinant AAV packaging function recombinant HSV.

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Improved electrochemical performances of polyaniline nanotubes-poly-L-lysine composite for label-free impedance detection of DNA hybridization

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Science china Chemistry 2010年第6期53卷 1371-1377页

作者： YANG Tao,JIANG Chen,ZHANG Wei & JIAO Kui key laboratory of Eco-chemical engineering,Ministry of Education College of Chemistry and molecular engineering,Qingdao University of Science and Technology,Qingdao 266042,china key laboratory of Eco-chemical engineering Ministry of Education College of Chemistry and molecular engineering Qingdao University of Science and Technology Qingdao China

A sensitive label-free DNA hybridization biosensing platform was fabricated based on the synergistic effect of polyaniline nanotubes (PANInt) and poly-L-lysine (pLys).The composite of pLys and PANInt was coated onto the carbon paste electrode (CPE) to form a uniform and very stable nanocomposite *** pLys in the composite film not only acts as a membrane to retain good electron transfer capability of PANInt even at physiological pH,but also possesses fine biocompatibility for *** probes with negatively charged phosphate groups were readily linked to the positively charged pLys surface due to the strong electrostatic *** synergistic effect of PANInt and pLys could significantly enhance the sensitivity of DNA hybridization *** phosphinothricin acetyltransferase (PAT) gene fragment from transgenic corn and the polymerase chain reaction amplification of the terminator of nopaline synthase gene from the real sample of a kind of transgenic soybean were detected by this DNA electrochemical biosensor via label-free impedance *** stable composite gives convenient permselectivity properties as a transducer material for the design of modern electrochemical impedance biosensor using [Fe(CN)6]3-/4as an indicator.

关键词： poly-L-lysine polyaniline nanotubes DNA electrochemical biosensor PAT gene NOS gene

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AtbHLH29 of Arabidopsis thaliana is a functional ortholog of tomato FER involved in controlling iron acquisition in strategy I plants

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Cell Research 2005年第8期15卷 613-621页

作者： You Xi YUAN1,2, Juan ZHANG1,2, Dao Wen WANG1, Hong Qing LING1,* 1The State key laboratory of Plant Cell and Chromosome engineering, Institute of genetics and Developmental biology, Chinese Academy of Sciences, Datun Road, Chaoyang District, beijing 100101, china 2Graduate School of the Chinese Academy of Sciences, Yuquan Road, Shijingshan District, beijing 100039, china The State key laboratory of Plant Cell and Chromosome engineering Institute of Genetics and Developmental Biology Chinese Academy of Sciences Beijing China Graduate School of the Chinese Academy of Sciences Beijing China

AtbHLH29 of Arabidopsis, encoding a bHLH protein, reveals a high similarity to the tomato FER which is proposed as a transcriptional regulator involved in controlling the iron deficiency responses and the iron uptake in tomato. For identification of its biological functions, AtbHLH29 was introduced into the genome of the tomato FER mutant T3238fer mediated by Agrobacterium tumefaciencs. Transgenic plants were regenerated and the stable integration of AtbHLH29 into their genomes was confirmed by Southern hybridization. molecular analysis demonstrated that expression of the exogenous AtbHLH29 of Arabidopsis in roots of the FER mutant T3238fer enabled to complement the defect functions of FER. The transgenic plants regained the ability to activate the whole iron deficiency responses and showed normal growth as the wild type under iron-limiting stress. Our transformation data demonstrate that AtbHLH29 is a functional ortholog of the tomato FER and can completely replace FER in controlling the effective iron acquisition in tomato. Except of iron, FER protein was directly or indirectly involved in manganese homeostasis due to that loss functions of FER in T3238fer resulted in strong reduction of Mn content in leaves and the defect function on Mn accumulation in leaves was complemented by expression of AtbHLH29 in the transgenic plants. Identification of the similar biological functions of FER and AtbHLH29, which isolated from two systematically wide-diverged “strategy I” plants, suggests that FER might be a universal gene presented in all strategy I plants in controlling effective iron acquisition system in roots.

关键词： tomato AtbHLH29 iron uptake Arabidopsis FER plant nutrition.

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Stacking interaction in metal complexes with compositions of DNA and heteroaromatic N-bases

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Chinese Science Bulletin 2002年第1期47卷 1-9页

作者： Jl Liangnian & LE XueyiDepartment of Chemistry, key laboratory for the gene engineering of the Ministry of Education of china, and State key laboratory of Ultrafast Laser Spectroscopy, Zhongshan University, Guangzhou 510275, china Department of Applied C Department of Chemistry Key Laboratory for the Gene Engineering of the Ministry of Education of China and State Key Laboratory of Ultrafast Laser Spectroscopy Zhongshan University Guangzhou China Department of Applied Chemistry South-China Agricultural University Guangzhou China

The current development in the intramolecular aromatic-ring stacking interactions in the complexes with compositions of DNA and heteroaromatic N-bases has been reviewed to a great extent, especially the significant contributions in several important systems about ternary mixed-ligand complexes, including nucleotide-metal ion-po-lyaromatic amine, amino acid-metal ion-polyaromatic amine, nucleotide-metal ion-pyridine-like aromatic amine, nucleo-tide-metal ion-amino acid, nucleotide-metal ion-nucleic acid base, nucleic acid base-metal ion, and the important factors affecting the intramolecular aromatic-ring stacking interac-tions in the complexes. Based on the study of stacking inter-action in the complexes, the mechanism of interaction be-tween DNA molecules and complexes of heteroaromatic N-bases has been established, which is crucial for the design and synthesis of the complexes acting as molecular devices of DNA.

关键词： ternary mixed-ligand complexes stacking interaction transition metal ion heteroaromatic N-base nucleotide amino acid.

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