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Functional Analysis of Multiple Transcription Factor Sites in a Regulatory Element of Human ε-Globin Gene

Acta Biochimica et Biophysica Sinica 2004年第10期36卷 673-680页

作者： Chun-Hui HOU, Jian HUANG, and Ruo-Lan QIAN* State Key Laboratory of Molecular Biology, 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 State Key Laboratory of Molecular Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of Sciences

The developmental control of the human ε-globin gene expression is mediated by transcrip- tional regulatory elements in the 5’ flanking DNA of this gene. A previously identified negative regulatory element (–3028 to –2902 bp, termed ε-NRAII) was analyzed and one putative NF-κB site and two GATA sites locate at –3004 bp, –2975 bp and –2948 bp were characterized. Electrophoresis mobility shift assay (EMSA) showed that the putative NF-κB site was specifically bound by nuclear proteins of K562 cells. Data obtained from transient transfection showed that the expression of reporter gene could be upregulated about 50% or 100% respectively when ε-NRAII was inserted upstream of the SV40 promoter or ε-globin gene proximal promoter (–177 bp to +1 bp), suggesting that ε-NRAII might not be a classic silencer. Mutation in the putative NF-κB site or in the GATA site (at –2975 bp) slightly reduced the expression of reporter gene driven by SV40 promoter or ε-globin gene proximal promoter. However, the mutation of GATA site at –2948 bp remarkably reduced the reporter gene activity driven by SV40 promoter, but not by ε-globin gene proximal promoter. Further mutation analysis showed that the negative effect of mutation in GATA site at –2948 bp on SV40 promoter was not affected by the mutation of the putative NF-κB site, whereas it could be abolished by the mutation of GATA site at –2975 bp. Furthermore, the mutation of both GATA sites could synergistically reduce the reporter gene activity driven by ε-globin gene proximal promoter. Those results suggested that ε- NRAII might function differently on the SV40 promoter and ε-globin gene proximal promoter.

关键词： ε-globin gene regulatory element NF-κB GATA

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An Effective Method for Raising Antisera Against β-defensins: Double-copy Protein Expression of mBin1b in E. coli

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Acta Biochimica et Biophysica Sinica 2004年第8期36卷 571-576页

作者： Li-Qing XIAO, Ai-Hua LIU, and Yong-Lian ZHANG State Key Laboratory of Molecular Biology, 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 State Key Laboratory of Molecular Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of Sciences

Bin1b is a rat epididymis specific β-defensin which may have fertility related functions in addition to its antimicrobial activity. β-defensins are cysteine-rich cationic antimicrobial peptides that have their important implications in innate and adaptive immunity. Though considerable numbers of new β-defensins have been discovered, few corresponding antibodies have been reported. The small peptide with special structure and antimicrobial nature of β-defensins make them very difficult to express in prokaryotic system. Here we adopted a double-copy protein expression scheme based on which not only the mBin1b protein was successfully expressed but also the immunity of the antigen was enhanced. The validity of the antisera was verified by using Western blotting and immunohistochemical analyses. It will be a useful tool for deeply investigating the roles of Bin1b and also provide a simple but effective method in raising antisera against other members of the β-defensin gene family.

关键词： Bin1b epididymis β-defensin double-copy protein polyclonal antiserum

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Progress in glial cell studies in some laboratories in china

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Science china(Life sciences) 2010年第3期53卷 330-337页

作者： DUAN ShuMin institute of Neuroscience,State Key Laboratory of Neuroscience,shanghai institutes for Biological sciences,chinese academy of sciences,shanghai china,200031 1. institute of Neuroscience State Key Laboratory of Neuroscience Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China 200031

Glial cells in the central nervous system(CNS) consist of a heterogeneous population of cell types,each characterized by distinct morphological features,physiological properties,and specific *** contrast to the previous view that glial cells were passive elements in the brain,accumulating evidence suggests that glial cells are active participants in various brain functions and brain *** review summarizes recent progress of glial cell studies from several groups in china,ranging from studies about the mechanisms of neuron-glia crosstalking to investigations on the roles of glial cells in various CNS disorders.

关键词： neuron-glia interaction ATP plasticity pain ischemia neural injury and repair

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Synapse formation and remodeling

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Science china(Life sciences) 2010年第3期53卷 315-321页

作者： LUO ZhenGe institute of Neuroscience and State Key Laboratory of Neuroscience,shanghai institutes for Biological sciences,chinese academy of sciences,shanghai 200031,china 1. institute of Neuroscience and State Key Laboratory of Neuroscience Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China

Synapses are specialized structures that mediate information flow between neurons and target cells,and thus are the basis for neuronal system to execute various functions,including learning and *** are around 1011 neurons in the human brain,with each neuron receiving thousands of synaptic inputs,either excitatory or inhibitory.A synapse is an asymmetric structure that is composed of pre-synaptic axon terminals,synaptic cleft,and postsynaptic *** formation involves a number of cell adhesion molecules,extracellular factors,and intracellular signaling or structural *** the establishment of synaptic connections,synapses undergo structural or functional changes,known as synaptic plasticity which is believed to be regulated by neuronal activity and a variety of secreted *** review summarizes recent progress in the field of synapse development,with particular emphasis on the work carried out in china during the past 10 years(1999-2009).

关键词： synapse development neuronal activity synaptic plasticity

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Pain research in china

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Science china(Life sciences) 2010年第3期53卷 356-362页

作者： ZHANG Xu institute of Neuroscience,State Key Laboratory of Neuroscience,shanghai institutes for Biological sciences,chinese academy of sciences,shanghai 200031,china 1. institute of Neuroscience State Key Laboratory of Neuroscience Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China

In addition to investigating the anatomy,neurochemistry and neurophysiology of pain pathways,chinese researchers have extended their work into the molecular and cellular mechanisms of sensory afferent transmission at the spinal cord level as well as cognitive processing in the *** mechanism underlying acupuncture analgesia remains a subject of special interest for chinese pain researchers,with the aim of combining clinical practice with the understanding of pain transmission and analgesic mechanism.

关键词： dorsal root ganglion spinal cord nociception receptors ion channels gene expression

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Detection of Tumor Marker CA125 in Ovarian Carcinoma Using Quantum Dots

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Acta Biochimica et Biophysica Sinica 2004年第10期36卷 681-686页

作者： Hui-Zhi WANG, Hai-Yan WANG, Ru-Qiang LIANG, and Kang-Cheng RUAN* Key Laboratory of Proteomics, 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 Key Laboratory of Proteomics Institute of Biochemistry and Cell BiologyShanghai Institutes for Biological SciencesChinese Academy of SciencesGraduate School of the Chinese Academy of SciencesShanghai 200031China

Semiconductor quantum dots (QDs) offer several advantages over organic dyes in fluores- cence-imaging applications, such as higher quantum yield, exceptional photostability, and a narrow, tunable, and symmetric emission spectrum. To explore whether QDs could specifically and effectively label tumor markers and be used in immunohistochemistry as a novel type of fluorescent probe, we used quantum dots with maximum emission wavelength 605 nm (QD605) to detect the ovarian carcinoma marker CA125 in specimens of different types (fixed cells, tissue sections, and xenograft piece). Additionally, we compared the photostability of QD signals with that of a conventional organic dye, FITC. All labeling signals of QDs were found to be more specific and brighter than those of FITC. Moreover, the QDs exhibited exceptional photostability during continuous illumination for 1 h by a high-intensity laser (Ar laser power 100 mW) at 488 nm, while the FITC signals faded very quickly and became undetectable after 24 min of illumination. These results indicate that QD-based probes can offer substantial advantages over existing fluorophores in many applications, and can be used effectively in immunohistochemistry as a novel class of fluorescent probes.

关键词： quantum dots detection of tumor marker CA125 ovarian carcinoma

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Nucleic Acid Binding Activity of Pns6 Encoded by Genome Segment 6 of Rice Ragged Stunt Oryzavirus

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Acta Biochimica et Biophysica Sinica 2004年第7期36卷 457-466页

作者： Chao-Gang SHAO#, Hui-Juan Lü#, Jian-Hua WU, and Zu-Xun GONG* Key Laboratory of Proteomics, institute of Biochemistry and Cell Biology, shanghai institutes of Life sciences, chinese academy of sciences, Graduate School of the chinese academy of sciences, shanghai 200031, china Key Laboratory of Proteomics Institute of Biochemistry and Cell Biology Shanghai Institutes of Life Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of Sciences Shanghai 200031 China

The ORF of genome segment 6 (S6) of rice ragged stunt oryzavirus (RRSV) Philippines isolate was cloned and sequenced based on the S6 sequence of the Thailand isolate. Pns6, the 71 kD product of S6 expressed in E. coli, was demonstrated to be a viral non-structural protein of RRSV by Western blotting. The gel mobility shift assays showed that Pns6 had nucleic acid binding activity. Pns6 could interact with single- and double-stranded forms of DNA and RNA, showing a preference for single-stranded nucleic acid and a slight preference for RRSV ssRNA over the rice ssRNA, as demonstrated by both competition and displacement assays. The binding of Pns6 to nucleic acids is strong and sequence non-specific. By using five truncated derivatives of Pns6, it was found that the basic region from amino acid 201 to 273 of Pns6 was the unique nucleic acid binding domain. Subcellular fractionation of leaf tissues of RRSV-infected rice plants and subsequent Western blotting had shown that Pns6 accumulated predominately in the cytoplasmic membrane fraction. The possible role of RRSV Pns6 in virus replication and assembly is discussed.

关键词： rice ragged stunt oryzavirus (RRSV) nucleic acid binding non-structural protein S6

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X-ray Powder Diffraction Pattern of Bi_4(SiO_4)_3

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Journal of Materials Science & Technology 1997年第2期13卷 145-148页

作者： Hongchao LIU Changlin KUO (shanghai institute of Ceramics, chinese academy of sciences, shanghai 200050, china) shanghai institute of Ceramics Chinese Academy of Sciences Shanghai 200050 China

In cooperation with figure-of-merits the Rietveld analysis can appraise both angular and intensity data of powder diffraction. In this work, X-ray diffraction pattern of Bi4(SiO4)3 was redetermined with intensity figure-of-merits, which qualify agreement between observed and calculated relative intensities. F30 is 158.90 (0.0059, 32), intensity figure of merit Rint is 8.7, I20(17), 8.0. The values of figure-of-merits show that the data of JCPDS cards are distorted. Both the experimental and calculated peak positions and heights are listed in detail.

关键词： SiO4 X-ray Powder Diffraction Pattern of Bi4 Bi

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Valence Distribution in AT_xV_(6-X)O_(11) Compounds

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Journal of Materials Science & Technology 1999年第6期15卷 551-554页

作者： Hongchao LIU and Changlin KUO(shanghai institute of Ceramics, chinese academy of sciences, shanghai 200050, china) chinese Acad Sci Shanghai Inst Ceram Shanghai 200050 Peoples R China

The distribution of trivalent and tetravalent cations in several ATxV6-xO11 compounds was quantitatively studied by the bond valence method. In SrV6O11, the M(3) sites were mainly occupied by trivaient cations; while M(1) and M(2) sites were shared by trivalent and tetravalent cations, the relative content of tetravalent at M(1) sites was higher than at M(2) sites. During substitution process, the trivalent cations preferred to occupy M(3) sites, tetravalent ones preferred to occupy M(2) sites. The occupancy of trivalent and tetravalent cations at M sites would change with the substitution

关键词： AT X)O Valence Distribution in AT_xV Compounds

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Directional migration of leukocytes: their pathological roles in inflammation and strategies for development of anti-inflammatory therapies

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Cell Research 2001年第2期11卷 85-88页

作者： GENG JIAN Guo institute of Biochemistry and Cell Biology, shanghai institutes for Biological sciences, chinese academy of sciences, 320 Yue Yang Road, shanghai 200031, china institute of Biochemistry and Cell Biology Chinese Academy of Sciences Shanghai. jggeng@***

Directional migration of leukocytes is indispensable to innate immunity for host defense. However, recruitment of leukocytes to a site of tissue injury the constitutes a leading cause for inflammatory responses. Mechanistically, it involves a cascade of cellular events precisely regulated by temporal and spatial present tation of a repertoire of molecules in the migrating leukocytes and their surroundings (microenvironments). Here I will summarize the emerging evidence that has shed lights on the underlying molecular mechanism for directional migration of leukocytes, which has guided the therapeutical development for innovative anti-inflammatory medicines.

关键词： Directional cell migration leukocytes inflammation therapy.

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