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The Effect of CD3-Specific Monoclonal Antibody on Treating Experimental Autoimmune Myasthenia Gravis

Cellular & Molecular Immunology 2005年第6期2卷 461-465页

作者： Ruonan Xu~1 Jianan Wang~1 Guojiang Chen~1 Gencheng Han~1 Renxi Wang~1 Beifen Shen~1 Yan Li~(1,2) ~1institute of basic medical science,Academy of military medical science,Beijing 100850,China ~2Department of Molecular Immunology,institute of basic medical science,27 Taiping Road,Beijing 100850,China. institute of basic medical science Academy of Military Medical Science Beijing 100850 China.

CD3-specific monoclonal antibody was the first one used for clinical practice in field of transplantation. Recently, renewed interests have elicited in its capacity to prevent autoimmune diabetes by inducing immune tolerance. In this study, we tested whether this antibody can also be used to treat another kind of autoimmune disease myasthenia gravis （MG） and explored the possible mechanisms. MG is caused by an autoimmune damage mediated by antibody- and complement-mediated destruction of AChR at the neuromuscular junction. We found that administration of CD3-specific antibody （Fab）2 to an animal model with experimental autoimmune myasthenia gravis （EAMG）（B6 mice received 3 times of AChR/CFA immunization） could not significantly improve the clinical signs and clinical score. When the possible mechanisms were tested, we found that CD3 antibody treatment slightly down-regulated the T-cell response to AChR, modestly up-regulation the muscle strength. And no significant difference in the titers of IgG2b was found between CD3 antibody treated and control groups. These data indicated that CD3-specific antibody was not suitable for treating MG, an antibody- and complementmediated autoimmune disease, after this disease has been established. The role of CD3-specific antibody in treating this kind of disease remains to be determined. Cellular ＆ Molecular Immunology. 2005;2（6）：461-465.

关键词： CD3-specific monoclonal antibody immune tolerance EAMG

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Disruption of estrogen receptor beta in mice brain results in pathological alterations resembling Alzheimer disease

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Acta Pharmacologica Sinica 2004年第4期25卷 54-59页

作者： Qing-hong ZHANG2, Yan-hong HUANG3, Yu-zhen HU, Geng-ze WEI,Xue-feng HAN, Shun-yan LU, Yu-feng ZHAO 2Department of Physiology, institute of basic medical science, Departmentof Gynaecology and obstetrics, 3 Xijing Hospital, The Fourth Military medicalUniversity, Xi-an 710032, Shaanxi, China Department of Physiology Institute of Basic Medical Science Department of Gynaecology and obstetrics Xijing Hospital The Fourth Military Medical University

AIM: To study the pathological characteristics of the mice with estrogen receptor β (ERβ) disruption in brain. METHODS: Immunohistochemistry method was applied in the study. RESULTS: β-Amyloid peptide(Aβ42) and apolipoprotein E (ApoE) immunoreactive substances were accumulated notably in cortex and limbic structures such as the hippocampus and amygdala in brain, resembling the pathological changes of human Alzheimer disease (AD). Aβ formed cloudy-like deposits in parenchyma of brain, while apoE also deposited along or surrounding the blood vessels. CONCLUSIONS: ERβ is crucial to the development of neural degenerative disease, so modulation of Aβ metabolism via ERβ signal pathway might be beneficial for AD prevention or therapy.

关键词： beta-amyloid peptide apolipoprotein E estrogen receptor beta Alzheimer disease

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Molecular characterization of human reticulon 3 as a potential marker during the differentiation of human neuroblastoma SH-SY5Y cells

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Progress in Natural science:Materials International 2004年第7期 90-93页

作者： YANG Hongbo 1, HUANG Xiaowei 1, YIN Bin 1, PENG Xiaozhong 1, FAN Ming 2, FAN Wenhong 2, QIANG Boqin 1 and YUAN Jiangang 1*(1. National Laboratory of medical Molecular Biology, institute of basic medical sciences, Chinese Academy of medical sciences, Peking Union medical College, Beijing 100005, China 2. Department of Neurobiology, institute of basic medical science, Academy of Military medical sciences, Beijing 100850, China)

Neuroendocrine-specific protein (NSP) -reticulons are endoplasmic reticulum-associated protein complexes, which are localized in the endoplasmic reticulum (ER) and identified as markers for neuroendocrine differentiation. In the present study, human reticulon 3 gene (hRTN3) was cloned and its expression pattern in a variety of tissues was investigated. Truncated hRTN3s corresponding to the C-terminal (hRTN3-C) domain were expressed and purified. hRTN3 mRNA was down-regulated during the differentiation of human neuroblastoma cell line SH-SY5Y induced by all-trans-retinoic acid (RA), which suggests that, like other members of the reticulon family, hRTN3 is a potential marker for neuroendocrine differentiation.

关键词： human reticulon 3 expression differentiation.

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Effect of P145^(INK4b/MTS2) on the proliferation of human hepatoma cells SMMC-7721

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Chinese science Bulletin 2000年第15期45卷 1408-1412页

作者： ZHANG Hong, LIU Huitu & SHI Fawu1. Key Laboratory of Cell Proliferation and Regulation Biology, Beijing Normal University, Beijing 100875, China 2. institute of basic medical science, General Hospital of People’s Liberation Army LA, Beijing 100039, China 3. institute of medical Information, medical Academy of China, Beijing 100020, China Correspondence should be addressed to Liu Huitu Key Laboratory of Cetl Proliferation and Regulation Biology Beijing Normal University Beijing China institute of medical Information Medical Academy of China Beijing China institute of basic medical science General Hospital of People’s Liberation Army LA Beijing China

The full length cDNA coding for P15 INK4b, which is a cyclin-dependent kinase inhibitor, was cloned to plasmid PXJ41-neo (Eco R I IXho 1 site) and the new constructed plasmid pXJp15 was obtained. pXJp15 was transferred into the human hepatoma SMMC-7721 cells by lipofectine reagent. After G418 selection, a series of cell lines stably expressing high levels of P15 (named SHT) and the clone containing vector PXJ41-neo only (named SVXJ) were obtained by Northern and Western analysis. The results showed that the proliferation of SHT cells is inhibited compared with that of SVXJ cells. Cell cycle analysis indicated that overexpressing of P15 inhibited the growth of SHT cells by decreasing progrssion of cells from G1 to S and G2 to M phases. The levels of c-Myc and c-Fos were obviously decreased in SHT cells compared with control cells by Western blotting. The decreased expression of oncogene may be one of the molecular mechanisms of the effect of P15 on the proliferation of in SHT cells.

关键词： CKIP15 human hepatoma cell cycle oncogene.

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Human nectin-like 1, a novel membrane protein interacting with protein 4.1 R

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Progress in Natural science:Materials International 2005年第2期 94-97页

作者： YANG Hongbo1, ZHOU Yan1, HU Xiaoyan1, XU Yaqin1, YIN Bin1, ZHU Shengtao1, FAN Wenhong2, FAN Ming2, QIANG Boqin1, YUAN Jiangang1 and PENG Xiaozhong1*(1. National Laboratory of medical Molecular Biology, institute of basic medical sciences, Chinese Academy of medical sciences, Peking Union medical College, Beijing 100005, China 2. Department of Neurobiology, institute of basic medical science, Academy of Military medical sciences, Beijing 100850, China) National Laboratory of medical Molecular Biology Institute of Basic Medical SciencesChinese Academy of Medical SciencesPeking Union Medical CollegeBeijing 100005China Department of Neurobiology Institute of Basic Medical ScienceAcademy of Military Medical SciencesBeijing 100850China

Human nectin-like 1 (NECL1) full-length cDNA was cloned by bioinformatics method when searching for candidate membrane proteins interacting with members of protein 4.1 family. The cytoplasmic and extracellular regions of NECL1 were expressed in and purified from ***, and the polyclonal antibody was produced. Interaction between the cytoplasmic region of NECL1 and the 30 kD membrane binding domain of protein 4.1 on red blood cell (4.1R) was demonstrated by IAsys-biosensor system and GST pull-down experiment. Results of biotin-labeled peptide ELISA further demonstrated the key amino acids for the binding. The interaction research of NECL1's cytoplasmic domain provides basis for further study of the functions of NECL1 in nervous system.

关键词： nectin-like 1 protein-protein interaction protein 4.1 R.

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噬菌体肽库中筛选NMDA受体抗原表位

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生物化学与生物物理进展 2003年第4期30卷 599-604页

作者：康晓楠孙长凯范明薛沿宁邵宁生赵杰韩大跃施广霞大连医科大学脑疾病研究所大连116027 军事医学科学院基础医学研究所北京100850 解放军第210医院大连116021 大连医科大学病理生理学教研室大连116027

为了获得人N 甲基 D 天冬氨酸受体 (NR)主亚基 (NR1)M3 M4环B细胞表位 ,以人NR1分子M3 M4环单克隆抗体MAB36 3淘筛噬菌体展示随机 12肽库 ,对筛选克隆进行特异性ELISA检测和竞争结合实验分析 .从 30个克隆中得到一个阳性克隆序列“VHTN... 详细信息

为了获得人N 甲基 D 天冬氨酸受体 (NR)主亚基 (NR1)M3 M4环B细胞表位 ,以人NR1分子M3 M4环单克隆抗体MAB36 3淘筛噬菌体展示随机 12肽库 ,对筛选克隆进行特异性ELISA检测和竞争结合实验分析 .从 30个克隆中得到一个阳性克隆序列“VHTNPSTWQPIL” (克隆 1) ,原核表达的NR1M 3 M 4环可以与克隆 1竞争结合MAB36 3.固相合成 5个表位探针短肽 ,发现其中只有R (2 2 )LRNPSKD可以与M3 M4环竞争结合抗体 ,将NPS三个氨基酸残基逐一敲除 ,缺失N或NP的合成肽和M 3 M 4环竞争抗体的能力减弱 ,缺失NPS的合成肽完全没有竞争能力 ,证明MAB36 3的表位为NPS ,S可能是关键性残基 .

关键词：噬菌体肽库 NMDA受体抗原表位 N-甲基-D天冬氨酸受体

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Evoked catecholamine release from sympathetic nerves in rodent cardiac slices

Evoked catecholamine release from sympathetic nerves in rode...

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中国生理学会第23届全国会员代表大会暨生理学学术大会

作者： J LV ~(1,2),Y XIU~(1,2),Y MU~(1,3),S GUO ~(1,2),XY ZHANG~1,Q LEI~(1,3),JL GU~(1,2),XJ KANG~1,CX ZHANG~1,LH LI~3,JM CAO~2,ZZHOU~1 1 institute of Molecular Medicine,Peking University,Beijing 100871,China 2 institute of basic medical science 3 Fuwai Hospital, Peking Union medical College,Beijing 100005,China

Neurotransmitter release from presynaptic cells can be recorded by postsynaptic potentials/currents in central nerve ***,little is known about how to record nerve release in cardiovascular system in fresh tissue or in *** have developed two novel methods for recording catecholamine release induced by nerve action potentials in cardiac slices and in vivo,*** beating is regulated by peripheral(vegas and sympathetic) nerves,which innervate and release neurotransmitters in the *** are interested in how nerves regulate heart *** the present study,we demonstrate first recordings from fresh cardiac *** have developed a novel method(termed cSEC) to record catecholamine release from rat and mouse sinus atrial nodes (SAN).Using micro carbon fiber electrode(CFE),catecholamine release from sympathetic nerve terminals were recorded as amperometric current(I) following depolarization made by field electric stimulation,high KCl,ACh, or *** spatial mapping of evoked catecholamine signals is determined in SAN.

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Evoked catecholamine release from sympathetic nerves in venticular slices

Evoked catecholamine release from sympathetic nerves in vent...

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中国生理学会第九届全国青年生理学工作者学术会议

作者： Shu Guo~(1,2),L(u|¨) J~(1,2),Xiu Y~(1,2),Mu Y~(1,2),Zhang XY~1,Lei Q~(1,3),Gu JL~(1,2),Kang XJ~1, Zhang CX~1,Li LH~3,Cao JM~2,Zhou Z~1 (1 institute of Molecular Medicine,Peking University,Beijing 100871,China 2 institute of basic medical science,and 3 Fuwai Hospital,Peking Union medical College,Beijing 100005,China.)

Neurotransmitter release from presynaptic cells can be recorded by postsynaptic potentials/currents in central nerve ***,little is known about how to record nerve release in cardiovascular system in fresh tissue or in *** have developed a novel method for recording catecholamine release induced by nerve action potentials in cardiac slices. Heart beating is regulated by peripheral(vegas and sympathetic) nerves,which innervate and release neurotransmitters in the *** are interested in how nerves regulate heart *** the present study,we demonstrate first recordings from fresh cardiac *** have developed a novel method(termed cSEC) to record catecholamine release from rat and mouse ventricular *** micro carbon fiber electrode(CFE),catecholamine release from sympathetic nerve terminals were recorded as amperometric current(I) following depolarization made by field electric stimulation, high KCl,or low *** are applying cSEC to determine phenotypes of sympathetic release in genetic disease models.

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胶束薄层色谱法同时测定APC片中3组分含量

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沈阳药科大学学报 2001年第5期18卷 338-340页

作者：赵怀清王学娅杨丽程丽珠沈阳药科大学药学院辽宁沈阳110016 辽宁省基础医学研究所辽宁沈阳110005 辽宁电业中心医院药剂科辽宁沈阳110015

应用胶束薄层色谱法同时测定复方乙酰水杨酸片 (APC)中的阿司匹林 (A)、非那西汀 (P)和咖啡因 (C)的含量。在聚酰胺薄膜上 ,以 2 %SDS 乙腈 NaAc Na2 HPO4 ·1 2H2 O(3∶1∶1 )为展开剂 ,测定波长为 2 73nm ,单波长反射法锯齿扫描 ,结... 详细信息

应用胶束薄层色谱法同时测定复方乙酰水杨酸片 (APC)中的阿司匹林 (A)、非那西汀 (P)和咖啡因 (C)的含量。在聚酰胺薄膜上 ,以 2 %SDS 乙腈 NaAc Na2 HPO4 ·1 2H2 O(3∶1∶1 )为展开剂 ,测定波长为 2 73nm ,单波长反射法锯齿扫描 ,结果 ,点样量在 0 9～ 4 5 μg,A组分的点样量X(mi)与斑点积分面积的比值Y(Ai/As)呈良好的线性关系 ,其回归方程为 :Y =2 0 0 3 1X +0 81 5 9,r =0 9981 ,平均回收率为 1 0 1 3 % ,RSD 1 9% ;点样量在 0 6～ 3 0 μg,P组分的回归方程为 :Y =1 2 1 4 5X +0 4 1 0 6,r=0 9978,平均回收率为 99 2 % ,RSD 2 4 % ;点样量在 0 1 4～ 0 7μg,C组分的回归方程为 :Y =2 1 3 63X +0 64 5 2 ,r=0 9984 ,平均回收率为 :98 4 % ,RSD 3 1 %

关键词：胶束薄层色谱法阿司匹林非那西汀咖啡因 APC 复方乙酰水杨酸片

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Establishment of liver specific glucokinase gene knockout mice: a new animal model for screening anti-diabetic drugs

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Acta Pharmacologica Sinica 2004年第12期25卷 93-99页

作者： Ya-li ZHANG, Xiao-hong TAN2, Mei-fang XIAO, Hui LI, Yi-qing Mao, Xiao YANG2, Huan-ran TAN3 Department of Pharmacology, School of basic medical science, Peking University Health science Center, Beijing 100083 2institute of Biotechnology Academy ofMilitary medical science, Beijing 100071, China Department of Pharmacology School of Basic Medical Science Peking University Health Science Center Beijing 100083 China

AIM: To characterize the liver-specific role of glucokinase in maintaining glucose homeostasis and to create an animal model for diabetes. METHODS: We performed hepatocyte-specific gene knockout of glucokinase in mice using Cre-loxP gene targeting strategy. First, two directly repeated loxP sequences were inserted to flank the exon 9 and exon 10 of glucokinase in genomic DNA. To achieve this, linearized targeting vector was electroporated into ES cells. Then G418- and Gancyclovir-double-resistant clones were picked and screened by PCR analysis and the positives identified by PCR were confirmed by Southern blot. A targeted clone was selected for microinjection into C57BL/6J blastocysts and implanted into pseudopregnant FVB recipient. Chimeric mice and their offspring were analyzed by Southern blot. Then by intercrossing the Alb-Cre transgenic mice with mice containing a conditional gk allele, we obtained mice with liver-specific glucokinase gene knockout. RESULTS: Among 161 double resistant clones 4 were positive to PCR and Southern blot and only one was used for further experiments. Eventually we generated the liver specific glucokinase knockout mice. These mice showed increased glucose level with age and at the age of 6 weeks fasting blood glucose level was significantly higher than control and they also displayed impaired glucose tolerance. CONCLUSION: Our studies indicate that hepatic glucokinase plays an important role in glucose homeostasis and its deficiencies contribute to the development of diabetes. The liver glucokinase knock- out mouse is an ideal animal model for MODY2, and it also can be applied for screening anti-diabetic drugs.

关键词： animal disease model non insulin dependent diabetes mellitus type 2 diabetes mellitus glucokinase knockout mice

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