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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 Chinese Academy of Medical Sciences School of Basic Medicine Peking Union Medical College Beijing

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

Cloning of the full-length gene of activin receptor-interacting protein2a and characterization of its interaction with ActR Ⅱ A

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Progress in Natural Science:Materials International 2007年第2期17卷 172-176页

作者： CUI Xueling, TAI Guixiang, ZHANG Hongjun, FANG Lin and LIU Zhonghui (school of basic Medical sciences, Jilin University, Changchun 130021, China) school of basic Medical sciences Jilin University Changchun 130021 China

To investigate the mechanism of intracellular signal transduction mediated by activin receptors, the full-length gene encoding a novel activin receptor-interacting protein2a (ARIP2a) was identified from a mouse brain cDNA library. The sequences of ARIP2a and ARIP2, distribution of ARIP2a and ARIP2 mRNA in mouse tissues, and expression of ARIP2a and ARIP2 in activin-induced RAW264.7 cell were compared, and the interaction between ARIP2a and ActRIIA was confirmed. The sequence analysis revealed that the full-length gene of ARIP2a, which composed of 1008 bp and encoded 153 amino acid residues, shared high sequence identity with ARIP2 except the position of the 99th amino acid. RT-PCR assay showed that ARIP2a mRNA was highly expressed in brain, pituitary and testis, and moderately in pancreas and ovary, but undetectable in other tissues. Whereas, ARIP2 mRNA was widely distributed in all mouse tissues that we tested. Moreover, expression of ARIP2a mRNA was significantly decreased in activin-stimulated RAW264. 7 cells; however, the expression of ARIP2 mRNA was increased. Additionally, the interaction between ARIP2a and activin type IIA receptor (ActRIIA) was further demonstrated by mammalian two-hybrid assays and pull-down assays. Taken together, those results indicate that although ARIP2a is homologous to ARIP2, they are different in tissue distribution and responses to activin. ARIP2a could also interact with activin type II receptor as a novel member of ARIP family.

关键词： activin receptor-interacting protein tissue distribution.

Effects of retinoic acid on proliferation,phenotype and expression of cyclin-dependent kinase inhibitors in TGF-β1-stimulated rat hepatic stellate cells

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World Journal of Gastroenterology 2000年第6期6卷 819-823页

作者： Guang Cun Huang Jin Sheng Zhang Yue E Zhang Department of Pathology school of basic Medical sciences,Fudan University.Shanghai 200032,China Guang Cun Huang Jin Sheng Zhang Yue E Zhang Department of Pathology School of Basic Medical Sciences Fudan University Shanghai 200032 China Guang Cun Huang graduated from Medical Center of Fudan University (former Shanghai Medical University) in 1996 majoring in forensic medicine now assistant and master at the Department of Pathology School of Basic Medical Sciences Fudan University specialized in the study of hepatic pathology having 3 papers published.

AIM To study the molecular mechanisms ofretinoic acid(RA)on proliferation andexpression of cyclin-dependent kinase inhibitors(CKI),i.e.p16,p21 and p27 in cultured rathepatic stellate cells(HSC)stimulated withtransforming growth factor beta 1(TGF-β1).METHODS HSC were isolated from healthy ratlivers and *** stimulated with1 mg/L TGF-β1,subcultured HSC were treatedwith or without 1 nmol/L *** assay,immunocytochemistry(ICC)for p16,p21,p27and α-smooth muscle actin(α-SMA)protein,insitu hybridization(ISH)for retinoic acidreceptor beta 2(RAR-β2)and p16,p21 and p27mRNA and quantitative image analysis(partially)were *** RA inhibited HSC proliferation(41.50%,P<0.05),decreased the protein levelof α-SMA(55.09%,P<0.05),and induced HSCto express RAR-β2 *** addition,RAincreased the protein level of p16(218.75%,P<0.05)and induced p21 protein expression;meanwhile,p27 was undetectable by ICC in bothcontrol and RA-treated ***,RA hadno influence on the mRNA levels of p16,p21 orp27 as determined by *** Up-regulation of p16 and p21 on post-transcriptional level may contribule, in part to RA inhibition of TGF-β1-initiated rat HSC activation in vitro.

关键词： retinoic acid cyclindependent kinase inhibitor hepatic stellate cell cell culture transforming growth factor beta 1 liver fibrosis

Assembly and structure of protein phosphatase 2A

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Science China(Life sciences) 2009年第2期52卷 135-146页

作者： SHI Yi Gong Center for Structural Biology, Department of Biological sciences and Biotechnology, and school of medicine, Tsinghua University, Beijing 100084, China 1. Center for Structural Biology Department of Biological Sciences and Biotechnology and School of Medicine Tsinghua University Beijing 100084 China

Protein phosphatase 2A (PP2A) represents a conserved family of important protein serine/threonine phosphatases in species ranging from yeast to human. The PP2A core enzyme comprises a scaffold subunit and a catalytic subunit. The heterotrimeric PP2A holoenzyme consists of the core enzyme and a variable regulatory subunit. The catalytic subunit of PP2A is subject to reversible methylation, medi-ated by two conserved enzymes. Both the PP2A core and holoenzymes are regulated through interac-tion with a large number of cellular cofactors. Recent biochemical and structural investigation reveals critical insights into the assembly and function of the PP2A core enzyme as well as two families of holoenzyme. This review focuses on the molecular mechanisms revealed by these latest advances.

关键词： protein phosphorylation dephosphorylation PP2A structure mechanism

Preparation and Characterization for Bispecific Antibodies of Anti-CD3×Anti-Idiotype to BCellLym phocytic Leukem ia

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Journal of Huazhong University of Science and Technology(Medical sciences) 1999年第3期19卷 166-169页

作者： SHEN Guanxin , ZHANG Yue , ZHU Huifen , YANG Jing , WANG Xiaolin Department of Immunology, school of basic Medical sciences, Tongji Medical University, Wuhan 430030 Department of Immunology School of Basic Medical Sciences Tongji Medical University Wuhan 430030

Bispecific antibodies (BsAbs) of anti CD3×anti idiotype (Id) to B cell lymphocytic leukemia (CLL) were prepared by chemical conjugation and direct hybridization technique of hybridoma and hybridoma without screening markers. The specificity of BsAbs from culture supernatants or ascites was assayed by indirect ELISA and indirect immunoflurescence (IF). The results showed that BsAbs could specifically react with homologous serum IgM from patients with B CLL and cells carrying CD3 marker respectively. Cell combination test and LDH assay demonstrated that BsAb significantly increased the conjugate formation between lymphocyte activated kill (LAK) cells and Daudi cells, and enhanced the cytotoxic activity of LAK cells against Daudi cells.

关键词： B chronic lymphocytic leukemia bispecific antibody quadruma chemical conjugation

Kinetics of thymocyte developmental process in fetal and neonatal mice

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Cell Research 2003年第4期13卷 265-273页

作者： SHI YUN XIAO, YAN LI, WEI FENG CHENDepartment of Immunology, school of basic Medical sciences, Peking University Health Science Center, 38 Xueyuan Road, Beijing 100083, China 1Department of Immunology School of Basic Medical Sciences Peking University Health Science Center 38 Xueyuan Road Beijing 100083 China 1Department of Immunology School of Basic Medical Sciences Peking University Health Science Center 38 Xueyuan Road Beijing 100083 China 1Department of Immunology School of Basic Medical Sciences Peking University Health Science Center 38 Xueyuan Road Beijing 100083 China

Kinetics of thymocyte development in vivo during embryogenesis was pursued. The early development of thymocytes in the fetal and neonatal BALB/c mice was discontinuous, with four waves of cell proliferation occurring at fetal day (Fd) 14 to 17, Fd 18 to day (D) 1 after birth, D 2 to D 5 and D6 thereafter. The first three proliferation waves coincided with the generation of CD4/"CD8/" (DP), TCR+CD4/hiCD8-/lo (CD4 SP), and TCR+CD4-/loCD8int/hi (CD8 SP) thymocytes, respectively. The transition from DN to DP cells was further investigated and it was found out that there were two differential pathways via immature single positive (ISP) cells in the BALB/c mice, each functioning at different fetal ages. One is via TCR-CD4-CD8+ cells, occurring between Fd 15 and Fd 17 and the other is via TCR-CD4+CD8- cells, occurring from Fd 17 until birth. In contrast, the TCR-CD4-CD8+ pathway dominated overwhelmingly in the C57BL/6 mice. These findings shed new light on the hypothesis that the differential pathway preferenc

关键词： fetal thymus neonatal mice thymocyte ontogeny.

Uniqueness and local stability for the inverse scattering problem of determining the cavity

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Science China Mathematics 2005年第8期48卷 1113-1123页

作者： FENG Lixin & MA Fuming LMAM, school of Mathematical sciences, Peking University, Beijing 100871, China school of Mathematical sciences, Heilongjiang University, Harbin 150080, China school of Mathematical sciences, Jilin University, Changchun 130012, China 1. LMAM School of Mathematical Sciences Peking University 100871 Beijing China 2. school of Mathematical sciences Heilongjiang University 150080 Harbin China 3. school of Mathematical sciences Jilin University 130012 Changchun China

Considering a time-harmonic electromagnetic plane wave incident on an arbitrarily shaped open cavity embedded in infinite ground plane, the physical process is modelled by Maxwell's equations. We investigate the inverse problem of determining the shape of the open cavity from the information of the measured scattered field. Results on the uniqueness and the local stability of the inverse problem in the 2-dimensional TM (transverse magnetic) polarization are proved in this paper.

关键词： uniqueness, stability, Maxwell's equations, inverse scattering problem.

Inhibitory Effect of TCCE on CCl4-induced Overexpression of IL-6 in Acute Liver Injury

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Acta Biochimica et Biophysica Sinica 2004年第11期36卷 767-772页

作者： Jing GAO, Huan DOU, Xin-Hui TANG, Li-Zhi XU, Yi-Mei FAN, and Xiao-Ning ZHAOschool of medicine, school of Life sciences, Nanjing University, Nanjing 210093, China school of medicine School of Life Sciences Nanjing University Nanjing 210093 Chin

Terminalia catappa L. leaves have been shown to protect against acute liver injury produced by some hepatotoxicants, but the active components and mechanisms are not clear. This study was designed to characterize the protective effects of the chloroform fraction of the ethanol extract of T. catappa leaves (TCCE) against carbon tetrachloride (CCl4)-induced hepatotoxicity in mice, and analyze the changes in expression level of interleukin-6 (IL-6) in the process. It was found that TCCE pretreatment (10 or 30 mg/kg, ig) protected mice from CCl4 toxicity, as evidenced by the reversed alterations in serum alanine aminotrans-ferase (sALT) and serum aspartate aminotransferase (sAST) activities. Additionally liver tissues were subjected to RT-PCR, Western blot and immunohistochemistry to analyze changes in IL-6 expression. It was found that TCCE markedly suppressed the CCl4-induced over-transcription of IL-6 gene. Consistent with the result, the expression of IL-6 protein was also blocked by TCCE in CCl4-stimula

关键词： acute liver injury TCCE CCl4 IL-6 expression

A New Peptide with Membrane-permeable Function Derived from Human Circadian Proteins

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Acta Biochimica et Biophysica Sinica 2004年第9期36卷 629-636页

作者： Tao PENG, Ying-Hui LIU, Chun-Lei YANG, Chao-Min WAN, Yao-Qi WANG, and Zheng-Rong WANG* Department of Biomedical Engineering, school of basic medicine and Forensic, Sichuan University, Chengdu 610041, China Department of Biomedical Engineering School of Basic Medicine and ForensicSichuan UniversityChengdu 610041China

basic peptides such as human immunodeficiency virus type 1 (HIV-1) Tat-(48-60) and Droso- phila Antennapedia-(43-58) have been reported to have a membrane permeability and a carrier function for intracellular protein delivery. Based on the fluorescence microscopic observations of the vascular endothelial cells (ECV-304) and the primary cultured neuroglial cells, we found that human Clock protein DNA-binding peptide [residue 35–47, hClock-(35-47)] had a translocation activity very similar to Tat-(48-60). The cellular uptake of hClock-(35-47) increases with the increase of incubation time and concentration. The internalization effect at 4 °C was same as that at 37 °C. Internalization of hClock-(35-47) was saturable and could be inhibited by the excess of the other MPPs. Moreover, The uptake of these peptides were significantly inhibited in the presence of heparan sulfate. These results strongly suggested that the hClock-(35-47) shared a common or very similar internalization pathway with other MPPs. Furthermore, we injected rat through the common carotid artery with hClock-(35-47)-FITC peptide, and cryostat sections of the brain were prepared and observed using a fluorescence microscope. Result showed that the peptide had the ability to translocate through the blood-brain barrier. It is promising to provide a new safe carrier for the intracellular and encephalic treatment.

关键词： membrane-permeable peptide (MPP) cell membrane hClock DNA-binding peptide blood-brain barrier