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cDNA cloning,sequence analysis,and recombinant expression of akitonin beta,a C-type lectin-like protein from Agkistrodon acutus

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Acta Pharmacologica Sinica 2004年第03期25卷 118-123页

作者： Xiang-dong ZHA1,2.Jing LIU1,Kang-sen XU3 1 School of Life Science,University of Science and Technology,Hefei 2.002. School of Life Science,Anhui University,2.Hefei 2.0039 national institute for control of pharmaceuticals and biological products,beijing 100050,China 3 School of Life Science University of Science and Technology School of Life Science Anhui University national institute for control of pharmaceuticals and biological products Beijing 100050 China

AIM: To clone the cDNA of a new member of snake venom C-type lectin-like proteins, to study its structure- function relationships and to achieve its recombinant production. METHODS: PCR primers were designed based on the homology and cDNA was amplified by RT-PCR using total RNA from snake venom gland as the template. The PCR products were cloned into the plasmid pGEM-T and sequenced. The deduced protein sequence was analyzed with some bioinformatic programs. A recombinant expression plasmid was constructed using pBAD- TOPO as vector and transformed into *** TOP10 competent cells. RESULTS: A novel cDNA sequence encod- ing akitonin β was found and accepted by GenBank (accession number AF387100). Akitonin β consists of a typical carbohydrate recognition domain (CRD) of C-type lectins, and it is homologous with other snake venom C-type lectin-like proteins. It was predicted to be a platelet antagonist. Upon induction with arabinose rAkitonin β express- ing in E coli was achieved at a high level (superior to 150 mg/L). The recombinant fusion protein exhibited inhibitory activities on rat platelet aggregation in vitro. CONCLUSION: A new member of snake venom C-type lectin-like proteins was discovered and characterized, and an efficient recombinant expression system was estab- lished for its production.

关键词： snake venoms cDNA akitonin C-type lectin-like proteins recombinant proteins structure- activity relationship

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Chiral Separation of Esmolol and Terazosin by Cyclodextrinmodified Capillary Zone Electrophoresis

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Chinese Chemical Letters 1998年第6期9卷 565-568页

作者： Yi Yang Dong Xue Qin REN Ai Jin HUANG Yi Liang SUN Zeng Pei SUN(Department of Chendstry, Peking University, beijing 10087)(national {2. for the control of Pharmaceutical and biological products, beijing 100050) Peking Univ Dept Chem Beijing 100871 Peoples R China Natl Inst control Pharmaceut & Biol Prod Beijing 100050 Peoples R China

Cyclodextrin-modified CZE was applied to the chiral separation of two basic drugs, i.e., esmolol and terazosin. Selector screening and concentration optimization experiments were performed. Resolution 3.1 for esmolol, 1.2.for terazosin were achieved when using 60 mmol/L gamma-CD and 60 mmoL/L DM-P-CD, respectively, in 50 mmol/L pH 2.5 sodium phosphate buffer.

关键词： capillary zone electrophoresis chiral separation cyclodextrin esmolol terazosin

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Separation of chiral drugs with β-CD phosphate by capillary electrophoresis

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Chinese Chemical Letters 2000年第2期11卷 153-156页

作者： Xue Qin REN1 Xiao Dan SU1 Al Jin HUANG1 Yi Liang SUN1 Zeng Pei SUNZ2.(1 Department of Chemistry, Peking University, beijing 100871 2.national {2. for the control of pharmaceutical and biological products, beijing 100050) Peking Univ Dept Chem Beijing 100871 Peoples R China Natl Inst control Pharmaceut & Biol Prod Beijing 100050 Peoples R China

β-Cyclodextrin phosphate (β-CD-phosphate) was used as a selector for separating chiral drugs by capillary electrophoresis (CE). A solution comprising of 12. mmol/L Britton Robinson buffer (BRB) containing 10 mmol/L 0-CD phosphate with the pH adjusted to 7.0 was used as the background electrolyte (BGE), and a small amount of analyte was injected (600v/1s). Triethylamine, diethylamine, triethanolamine, diethanolamine, Tris added as modifier were compared. Isoprenaline, methoxamine, oxprenolol, practolol were successfully resolved.

关键词： Capillary electrophoresis chiral separation β-cyclodextrin-phosphate isoprenaline methoxamine oxprenolol practolol.

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Crystal and Molecular Structure of Quinide

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Chinese Journal of Structural Chemistry 1996年第5期15卷 400-403页

作者： WANG Gang-Li ZHAO Shu-Jie CHEN De-Chang(national institute for the {2. of Pharmaceutical and biological Product, beijing, 100050)LU Yang WU Nan ZHENG Qi-Tai(institute of Material Medical,the Chinese Academy of Medical Sciences, beijing, 100050) Natl. Inst. Contr. Pharmaceut./B.P. Beijing 100050 institute of Material Medical Chinese Academy of Medical Sciences Beijing 100050

A pure title compound was isolated from water soluble extraction of the fruits of Gardenia sootepensis Hutch.,and its crystal structure was determined by X-ray diffraction. The crystal, chemical formula C_7H_(10)O_5 and Mr 174. 15, is colorless,transparent and belongs to monoclinic system, space group P2.1. The crystal data are as follows:a=6.467(4), b=18.349(4), c=7. 041(4), β= 116. 2. (5)°,V=749.7(6), Z=4,Dx=1.543 g/cm￣3,F(000)=368,μ=0.08 mm-1,λ(MoKα) =0. 7106,R=0.063. It is weak hydrobonds that result in slight conformation difference between the intermolecular H-bond associated molecules.

关键词： gardenia sootepensis quinide crystal structure

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Partial sequencing of 5' non-coding region of 7 HGV strains isolated from different areas of China

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World Journal of Gastroenterology 1999年第5期5卷 432-434页

作者： WANG Xing-Tai ZHUANG Hui SONG Hai-Bo Ll He-Min ZHANG Hua-Yuan and YU Yang(national institute for the {2. of bormceutical and biologicalPriducts, beijing 100050, China)(Department of Microbiology, beijing Medical University, beijing100083, China)(L national institute for the {2. of Pharmaceutical and biological products Beijing 100050 China Department of Virology Department of Microbiology Beijing Medical University Beijing 100083 China Laboratory for Clinical Diagnosis Hefei 230000 Anhui Province China

INTRODUCTIONAlthough sensitive tests for detection of known hepatitis viruses are available, the etiology of 10%-15% post-transfusion and community-acquired hepatitis cases has remained undefined. It suggests the exis... 详细信息

关键词： hepatitis G virus polymerase chain reaction nucleotide sequence RNA, viral

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Granulocyte-macrophage colony-stimulating factor DNA prime-protein boost strategy to enhance efficacy of a recombinant pertussis DNA vac-cine

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Acta Pharmacologica Sinica 2006年第11期27卷 1487-1494页

作者： Qing-tian LI~(2.3,5)Yong-zhang ZHU~(2.3,5)Jia-you CHU~(2.6)Ke DONG~3 Ping HE~3 Chun-yan FENG~3 Bao-yu HU~3 Shu-min ZHANG~4 Xiao-kui GUO~(3,6)2.institute of Medical Biology,Peking Union Medical College and Chinese Academy of Medical Sciences,Kunming 650118,China 3 Department of Medical Microbiology and Parasitology,Shanghai Jiao Tong University School of Medicine,Shanghai 2.002.,China 4 Chinese national institute for the control of Pharmaceutical and biological products,beijing 100050,China instituteofMedicalBiology PekingUnionMedicalCollegeandChineseAcademyofMedicalSciencesKunming650118China DepartmentofMedicalMicrobiologyandParasitology ShanghaiJiaoTongUniversitySchoolofMedicineShanghai200025China

Aim:To investigate a new strategy to enhance the efficacy of a recombinantpertussis DNA *** strategy is co-injection with cytokine plasmids asprime,and boosted with purified homologous ***:A recombinantpertussis DNA vaccine containing the pertussis toxin subunit 1(PTS 1),fragmentsof the filamentous hemagglutinin(FHA)gene and pertactin(PRN)gene encodingfilamentous hemagglutinin and pertactin were ***/c mice were im-munized with several DNA vaccines and antigen-specific antibodies anti-PTS1,anti-PRN,anti-FHA,cytokines interleukin(IL)-10,IL-4,IFN-γ,TNF-α,and spleno-cyte-proliferation assay were used to describe immune ***:Therecombinant DNA vaccine could elicit similar immune responses in mice as that ofseparate plasmids encoding the 3 fragments,*** immunized withDNA and boosted with the corresponding protein elicited more antibodies thanthose that received DNA as *** particular,when the mice were co-immu-nized with murine granulocyte-macrophage colony-stimulating factor plasmidsand boosted with proteins,all 4 cytokines and the 3 antigen-specific antibodieswere significantly increased compared to the pVAX1 ***-PTS1,anti-FHA,IL-4 and TNF-α elicited in the colony stimulating factor(CSF)prime-proteinboost group showed significant increase compared to all the other ***:This prime and boost strategy has proven to be very useful in im-proving the immunogenicity of DNA vaccines against pertussis.

关键词： granulocyte-macrophage colony-stimulating factor pertussis DNA vaccines

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Plasmacytoid Dendritic Cells Act as the Most Competent Cell Type in Linking Antiviral Innate and Adaptive Immune Responses

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Cellular & Molecular Immunology 2005年第6期2卷 411-417页

作者： Zheng Zhang~1 Fu-Sheng Wang~(1,2. ~1Research Center of biological Therapy,beijing 302.Hospital,beijing 100039,China ~2.esearch Centre of biological Therapy,beijing institute of Infectious Diseases,beijing 302.Hospital,100 Xi Si Huan Middle Road,beijing 100039,China. Research Center of biological Therapy Beijing 302 Hospital Beijing 100039 China.

Appropriate in vivo control of plasmacytoid dendritic cell (pDC) recruitment and activation is a fundamentalrequirement for defense against viral *** this process,a pivotal event that influences the outcome ofviral infection is the production of high levels of type I interferon by *** particular,recent research findingsshowed that pDCs not only shape the nature of innate resistance,but are also responsible for the successfultransition from innate to adaptive immunity for viral *** addition,pDCs can differentiate into antigenpresenting cells that may regulate tolerance to a given ***,in a series of recent clinical studies,pDCs appeared to be defective in number and function in conditions of chronic viral diseases such as infected withHIV-1,HBV or ***-associated clinical antiviral therapy is also *** review describes researchfindings examining the functional and antiviral properties of in vivo pDC *** & MolecularImmunology.2.05;2.6):411-417.

关键词： virus plasmacytoid dendritic cell innate immunity adaptive immunity

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Effects of the CNTF-collagen gel-controlled delivery system on rat neural stem/progenitor cells behavior

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Science China(Life Sciences) 2010年第4期53卷 504-510页

作者： YANG ZhaoYang1, QIAO Hui2.& LI XiaoGuang1,3 1School of biological Science and Medical Engineering, Beihang University, beijing 100191, China 2.ublic Heath School, Ningxia Medical University, Yinchuan 750004, China 3beijing institute for Neuroscience, Capital Medical University, beijing 100069, China 1. School of biological Science and Medical Engineering Beihang University Beijing 100191 China2. Public Heath School Ningxia Medical University Yinchuan 750004 China3. Beijing Institute for Neuroscience Capital Medical University Beijing 100069 China

The injury of central nervous system (CNS) usually causes the cavity formation. Although transplantation of neural stem/precursor cells (NSPCs) into the lesioned area of CNS has been shown to be implicated in the functional restoration, the therapeutic result is limited by the poor survival of NSPCs as well as their insufficient proliferation and differentiation abilities. Type-1 collagen is considered as a candidate scaffold or drug delivery system to overcome the aforementioned obstacle. This study observed the effects of the CNTF (ciliary neurotrophic factor)-collagen gel-controlled delivery system and daily addition of soluble-form CNTF on the NSPC survival, migration, proliferation and differentiation. The results showed that, within 12.h of the initial co-culture, CNTF was released in a burst pattern, then the CNTF-collagen gel-controlled delivery system stably released CNTF for up to 12.d. The cell viability test, together with immunohistochemistry, RT-PCR and Western blotting, showed that the CNTF-collagen gel-controlled delivery system supported the NSPCs seeded on the surface of collagen gel survival and facilitated their migration and proliferation. The daily addition of soluble-form CNTF to the medium had similar effects to the CNTF-collagen gel-controlled delivery system, but large quantities of soluble-form CNTF were consumed during the entire process. Taken together, the CNTF-collagen gel-controlled delivery system not only provides a physical scaffold for the transplanted NSPCs to adhere and migrate, but also facilitates the NSPC survival, growth and proliferation, simultaneously reducing the consumption of the expensive growth factors. This system may be used to enhance the microenvironment in the lesioned area of CNS.

关键词： spinal cord neural stem/progenitor cells control release type-1 collagen CNTF

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The Toxicity Evaluation of Recombinant Replicating Vaccinia Virus Vaccine in BALB/c Mice

The Toxicity Evaluation of Recombinant Replicating Vaccinia ...

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第十三届中国科协年会第5分会场-化学物的环境危害与健康风险评估国际研讨会

作者： Sanlong WANG 1 , Limei CAO 1 , Yin LIU 2., Danli DUAN 2., Xiuwen WANG 1 , Bo LI 1 , Yiming SHAO 2.(1. national Center for Safety Evaluation of Drugs, national {2. for the control of Pharmaceutical & biological products, Tianjin 300050, China 2. national Center for AIDS/STD Prevention and control, China CDC, beijing 100050, China)

To evaluate the preclinical safety of recombinant replicating vaccinia virus-based HIV vaccines (rVVV)in BALB/c mice. The study was composed of groups 1, 2. 3, 4, wherein the groups 1 and 2.received only one priming immunization and the groups 3, 4 received prime-boost immunizations at a 6-week interval. The dose given was 50μl/skin of back/2.dosing point by intradermal injection approximately 2.2.10 6 PFU. Cellular responses were evaluated in spleenocytes after restimulation on the following basis: detection of IFN-γ producing CD8 + -positive T cells, and CD4 + /CD8 + T cells ratio by FACS assay. Humoral responses were evaluated in serum samples on the basis of specific anti-rVVV antibodies. A real-time quantitative PCR was employed for detection of biodistribution of rVVV in tissues and organs. The results showed that there were no significant toxic effects and damages to major organs in control and rVVV-treated animals. The results of FACS analysis assay for the group 4 that received prime-boost immunization showed cellular immune responses, as indicated by high IFN-γ production by CD8 + -positive T cells. There was no significant change for the CD4 + /CD8 + T cells ratio in control and rVVV-treated animals. Immunogenicity of rVVV given with prime-boost doses was stronger than that of rVVV given with a single priming dose in BALB/c mice. Following intradermal injection of rVVV in mice, all tissues were Q-PCR-negative on Day 35 after dosing. The study showed that there were no significant toxic effects and the rVVV could induce both humoral and cellular immune responses in BALB/c mice.

关键词： Toxicity evaluation Recombinant replicating vaccinia virus Vaccine BALB/c Mice

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人vasostatin的克隆、表达、纯化及活性检测

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生物化学与生物物理进展 2003年第3期30卷 447-452页

作者：李永红王军志饶春明张翊中国药品生物制品检定所北京100050

从成人肝脏cDNA文库中 ,PCR扩增得到人vasostatin基因编码区序列 ,将此序列插入原核表达载体pQE3 0进行表达 ,SDS 聚丙烯酰胺凝胶电泳 (SDS PAGE)测定表明产物以包涵体形式存在 ,表达量占菌体总蛋白量的 5 0 %以上 .包涵体洗涤后溶于 8m... 详细信息

从成人肝脏cDNA文库中 ,PCR扩增得到人vasostatin基因编码区序列 ,将此序列插入原核表达载体pQE3 0进行表达 ,SDS 聚丙烯酰胺凝胶电泳 (SDS PAGE)测定表明产物以包涵体形式存在 ,表达量占菌体总蛋白量的 5 0 %以上 .包涵体洗涤后溶于 8mol/L尿素溶液 ,在变性条件下通过镍氨三乙酸 (Ni NTA)金属螯合亲和层析柱进行纯化后 ,再经透析进行复性 .N端氨基酸序列、分子质量、等电点等理化指标的测定结果与理论值相符 .用内皮细胞增殖试验、内皮细胞迁移试验以及鸡胚尿囊膜血管生成试验等方法进行活性检测 ,证实复性的表达产物具有抑制内皮细胞增殖和迁移。

关键词：人vosostatin基因克隆表达纯化活性检测生物学活性抗癌药物血管生成抑制因子

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