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Mechanism for chronic pain generation

第十届全国针刺麻醉针刺镇痛及针刺调整效应学术研讨会

作者： Zhi Xiaoa.b,Shan Oua,c,Wen-Juan Hea,Yan-Dong Zhaoa,Xiao-Hong Liub, Huai-Zhen Ruana, a Department of Neurobiology,college of basic medical sciences,Third Military medical University,Chongqing 400038.China,bdepartment of physiology.college of basic medical sciences,Zunyi medical college,Zunyi,Guizhou 563003,China, cDepartment of Anesthesiology,General Hospital of Chengdu Military Command, Chengdu,Sichuan 610083,China

Extracellular ATP facilitates pain transmission at peripheral and spinal sites via the P2X receptors and the P2X3 subtype is an important candidate for this effect. Electroacupuncture(EA) has been clinically utilized to manage chronic *** this study,with neuropathic pain model of Sprague-Dawley(SD) rats,the P2X3 receptor protein level and expression location in the midbrain periaqueductal gray(PAG),a

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Chinese Science Bulletin 2000年第9期45卷 775-783页

作者： XIE YikuanInstitute of basic medical sciences, Peking Union medical college, Chinese Academy of medical sciences, Beijing 100005. Institute of basic medical sciences Peking Union Medical College Chinese Academy of Medical Sciences Beijing China

Neuropathic pain and the other abnormalities of sensation induced by axon injury or by peripheral nerve inflammation should result from functional compensations of the injured neurons during their regeneration. Ectopic distribution of proteins related to Na+, K+ and Ca2+ channels as well as of receptors on both membranes of injured axon and its cell body becomes a main pacemaker from which spontaneous ectopic afferent of primary sensatory neurons and crosstalk between neurons occur. Abnormal ectopic afferent activities lead to disorders of the sensation, such as hyperalgesia, allodynia, spontaneous pain and paraesthesia. Administration of some ion channel agents and/or α2-adrenergic blockers has shown efficiency in preventing neuropathic pain development and in relieving neuropathic pain.

关键词： chronic pain nerve injury ectopic firing ion channels sympathetic activity.

Cardiac protective role of a novel erythrocyte-derived depressing factor on rats and its Ca^2+ mechanism

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Chinese Science Bulletin 2003年第24期48卷 2710-2714页

作者： WANG Yutang, WEN Yunyi, MA Ning & SHI Lei Department of physiology, Institute of basic medical sciences, Chinese Academy of medical sciences, School of basic Medicine, Peking Union medical college, Beijing 100005, China Correspondence should be addressed to Wen Yunyi (e-mail: wenyunyi@ ***) Department of physiology Institute of Basic Medical Sciences School of Basic Medicine Beijing 100005 China

The cardiac protective role of a novel erythro-cyte-derived depressing factor (EDDF) on spontaneous hy-pertensive rats (SHR), calcium overload (CaO) rats and Wistar rats and its mechanism was evaluated. Mean artery pressure (MAP), heart rate (HR) and LVdp/dtmax were measured by physiological recorder. The effect of EDDF on the Ca2+-ATPase activity in myocardial sarcoplasmic reticu-lum (SR) of CaO rats was determined by inorganic phos-phate assay. Calcium transport in myocytes was measured by 45Ca2+ radioactive isotope measurement. The phosphoryla-tion levels of extracellular signal-regulated protein kinases (ERK1/2) in myocardial tissue of SHR and CaO rats were measured by Western blot method. And the ultrastructures of cardiac muscle cells were observed with the transmission electron microscope. The results indicated that EDDF could significantly decrease MAP, HR and LVdp/dtmax in a dose dependent manner (P < 0.05). It seems that the mechanism might relate with activating the Ca2+-APTase, enhancing the uptake and release of Ca2+ from SR (P < 0.05), decreasing the phosphorylation levels of ERK1/2 of myocytes (P < 0.01) and lightening the ultrastructural lesion of cardiac muscle cells. In CaO rats, the Ca2+-ATPase activity decreased clearly com-pared to control (64.99 7.16 vs 94.48 7.68 nmol·min-1 ·mg-1 protein, P < 0.01), while EDDF (100 mg/mL) could significantly increase the activity (87.93 ?9.54 vs 64.99 ?7.16, P < 0.05, n = 7). Both uptake and release rate of Ca2+ (祄ol 45Ca2+/g protein/min) from myocardial SR of CaO rats re-markably decreased compared to control (32.40 ?2.70 and 15.46 ?1.49 vs 61.09 ?10.89 and 25.47 ?4.29, P < 0.05); EDDF (100 mg/mL) could significantly stimulate their activi-ties (50.48 6.76 and 21.76 2.75 vs 32.40 2.70 and 15.46 1.49, P < 0.05). EDDF could evidently down-regulate the phosphorylation of ERK1/2 in myocardial tissue from SHR and CaO rats (P < 0.01), lighten the ultrastructural lesion of cardiac muscle cells of SHR as well. It is

关键词： Ca^2+ 钙离子肌质网 SR 心律 HR 红细胞血压

DNA-protein interaction at erythroid important regulatory elements of MEL cells bv in vivo footprinting

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Chinese Science Bulletin 2000年第12期45卷 1101-1108页

作者： Jl Xinjun, LIU Depei, XU Dongdong, LI Lei, WANG Jing & LIANG ZhiquanNational Laboratory of medical Molecular Biology, Institute of basic medical sciences, Chinese Academy of medical sciences, Faculty of basic medical sciences, Peking Union medical college, Beijing 100005, China National Laboratory of medical Molecular Biology Institute of Basic Medical Sciences Chinese Academy of Medical Sciences Faculty of Basic Medical Sciences Peking Union Medical College Beijing China

Using ligation-mediated PCR method to study the status of DNA-protein interaction at hypersensitive site 2 of locus control Region and βmaj promoter of MEL cell line before and after induction, MEL cell has been cultured and induced to differentiation by Hemin and DMSO, then the live cells have been treated with dimethyl sulfate. Ligation mediated PCR has been carried out following the chemical cleavage. The results demonstrate that before and after induction, the status of DNA-protein interaction at both hypersensitive site 2 and βmaj promoter change significantly, indicating that distal regulatory elements (locus control region, hypersensitive sites) as well as proximal regulatory elements (promoter, enhancer) of β-globin gene cluster participate in the regulation of developmental specificity.

关键词： β-globin gene HS LM-PCR in vivo footprinting MEL cell line.

Antibody gene therapy: an attractive approach for the treatment of cancers and other chronic diseases

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Cell Research 2007年第4期17卷 303-306页

作者： Dexian Zheng ~1National Laboratory of medical Molecular Biology,Institute of basic medical sciences,Chinese Academy of medical sciences and Peking Union medical college,Beijing 100005,China National Laboratory of medical Molecular Biology Institute of Basic Medical Sciences Chinese Academy of Medical Sciences and Peking Union Medical College Beijing China

Monoclonal antibodies(mAb)have been successfullyapplied to the treatment of chronic diseases,such as cancer,inflammation and immune *** the technicaladvances in antibody engineering,development of smallrecombinant antibody fragments as high affinity therapeu-tics with reduced immunogenicity has become under thespotlight.A popular format of the engineered recombinantantibody fragments is the single-chain fixed-variable(scFv)molecules,in which the VH and VL regions of the parentalantibody are jointed together by a polypeptide *** fragment retains the target specificity and antigen-binding affinity of the intact antibody,and can be geneti-cally designed and produced in large quantity by ectopicallyexpressing both VH and VL regions from a single cDNAin *** to its smaller size,the scFv molecule showsimproved pharmacokinetics in tumor penetration and isbetter tolerated by the host immune *** thesepotential advantages of using scFv molecules for immu-notherapy,especially for chronic diseases such as cancers,the treatment efficacy,however,is often compromised bythe rapid blood clearance and insufficient concentration ofthe infused scFv fragments at the target *** achievehigh local concentrations in the target tissues,large amountof the recombinant scFv proteins(hundreds milligrams or

关键词： gene an attractive approach for the treatment of cancers and other chronic diseases Antibody gene therapy

Influenza A virus H5N1 entry into host cells is through clathrin-dependent endocytosis

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Science China(Life sciences) 2009年第5期52卷 464-469页

作者： WANG HongLiang & JIANG ChengYu National Key Laboratory of medical Molecular Biology, Institute of basic medical sciences, Chinese Academy of medical sciences and Peking Union medical college, Beijing 100005, China National Key Laboratory of medical Molecular Biology Institute of Basic Medical Sciences Chinese Academy of Medical Sciences and Peking Union Medical College Beijing 100005 China.

Influenza A virus H5N1 presents a major threat to human health. The entry of influenza virus into host cells is believed to be mediated by hemagglutinin (HA), a virus surface glycoprotein that can bind terminal sialic acid residues on host cell glycoproteins and glycolipids. In this study, we elucidated the pathways through which H5N1 enters human lung carcinoma cell line A549. We first proved that H5N1 can enter A549 cells via endocytosis, as lysosomotropic agents, such as bafilomycin A1 and chloroquine, can rescue H5N1-induced A549 cell death. By using specific inhibitors, and siRNAs that target the clathrin pathway, we further found that H5N1 could enter A549 cells via clathrin-mediated endocytosis, while inhibitors targeting caveolae-mediated endocytosis could not inhibit H5N1 cell entry. These findings expand our understanding of H5N1 pathogenesis and provide new information for anti-viral drug research.

关键词： influenza A H5N1 endocytosis lysosomotropic agents clathrin

Establishment and Utilization of a Tetracycline-controlled Inducible RNA Interfering System to Repress Gene Expression in Chronic Myelogenous Leukemia Cells

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Acta Biochimica et Biophysica Sinica 2005年第12期37卷 851-856页

作者： Fan YANG Yun ZHANG Ying-Li CAO Shu-Hui WANG Li LIU Department of Microbiology and Etiology,Institute of basic medical .sic medical sciences. Chinese Academy q/’medical sciences and Peking Union medical college.Beijing 100005.China Department of Microbiology and Etiology Institute of Basic Medical Sciences Chinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100005 China

RNA interference (RNAi),a posttranscriptional gene silencing process mediated by small double-stranded RNA specifically complementary to the targeted transcript,has been used extensively in the developmentof novel therapeutic approaches against various human diseases including chronic myelogenous leukemia (CML).*** report the successful construction of a tetracycline-controlled siRNA in CML cell line K562.A K562cell line stably expressing the reverse tetracycline-controlled transactivator (rtTA) was constructed.A tetracy-cline responsive element(TRE) was integrated into the RNA polymerase Ⅲ promoter region of pBS/U6 thatwas used to drive specific siRNA to target the novel cytokine receptor-like factor 3 (CRLF3) gene,The resultsshow that rtTA was able to recognize the TRE to prevent siRNA-mediated exogenous and endogenous CRLF3gene ***,CRLF3-siRNA mediated gene repression could be induced in a dose-dependentmanner in the presence of ***,the inducible siRNAi system in K562 cells might be useful for thestudy of RNAi-mediated therapeutic approaches against CML.

关键词： inducible siRNA tetracycline reverse tetracycline-controlled transactivator U6 promoter CRLF3

Cloning and Characterization of a Novel Gene SRG-L Expressed in Late Stages of Mouse Spermatogenic Cells

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Acta Biochimica et Biophysica Sinica 2004年第5期36卷 315-322页

作者： Rui GUO, Peng-Peng MA, Jing MA, Ye-Hua GE, She-Pu XUE, and Dai-Shu HAN*Department of Cell Biology, Institute of basic medical sciences, Chinese Academy of medical sciences & Peking Union medical college,Beijing 100005, China Department of Cell Biology Institute of Basic Medical Sciences Chinese Academy of Medical Sciences & Peking Union Medical College Beijing 100005 China

A cDNA expressed specifically in late stages of mouse spermatogenic cells during spermato- genesis was cloned by using overlapping RT-PCR and RACE. The cDNA contained an open reading frame (ORF) of 2625 base pairs that encoded an 874 amino acids protein. Comparison analysis of amino acid sequence showed 91% and 80% identity to a rat homologue XP-226242 and a monkey homologue BAB63115 respectively. The expression of the mRNA was only observed in pachytene spermatocytes, round, and elongating spermatids. We named this gene as SRG-L (spermatogenesis related gene expressed in the late stages of spermatogenic cells, GenBank accession No. AY352586). The tissue-specific analysis showed that the SRG-L was highly expressed in spleen and testis. The results suggested that SRG-L might play special roles during spermatogenesis, particularly related to meiosis and spermiogenesis. Analysis of the amino acid sequence showed there was a coiled-coil region near the N-terminal region and rich phosphorylation sites, suggesting SRG-L might function as a transmembrane protein mediating signal transduction. This study also demonstrated that gene cloning by RT-PCR was applicable and convenient when its homologous gene was known.

关键词： spermatogenesis RT-PCR RACE gene cloning

Avian influenza H5N1:an update on molecular pathogenesis

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Science China(Life sciences) 2009年第5期52卷 459-463页

Avian influenza A virus constitutes a large threat to human health. Recent outbreaks of highly pathogenic avian influenza H5N1 virus in poultry and in humans have raised concerns that an influenza pandemic will occur in the near future. Transmission from avian species to humans remains sporadic, but the mortality associated with human infection is very high (about 62%). To date, there are no effective therapeutic drugs or a prophylactic vaccines available, which means that there is still a long way to go before we can eradicate or cure avian influenza. This review focuses on the molecular pathogenesis of avian influenza H5N1 virus infection. An understanding of the viral pathogenesis may facilitate the development of novel treatments or effective eradication of this fatal disease.

关键词： avian flu H5N1 acute lung injury