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Characterization of MR-1,a Novel Myofibrillogenesis Regulator in Human Muscle

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Acta Biochimica et Biophysica Sinica 2004年第6期36卷 412-418页

作者： Tian-Bo LI, Xiu-Hua LIU1, Shuang FENG, Yang HU, Wei-Xi YANG, Yue HAN1, Yi-Guang WANG*, and Li-Min GONG2.Institute of Medicinal Biotechnology, CAMS and PUMC, beijing 100050, china 1{2. of Pathophysiology, PLA general hospital, beijing 100853, china 2.2. of Cardiology, the university of Texas MD Anderson Cancer Center, Houston, TX 77030, USA Institute of Medicinal Biotechnology CAMS and PUMCBeijing 100050 China department of Pathophysiology PLA General HospitalBeijing 100853 China department of Cardiology the University of Texas MD Anderson Cancer CenterHouston TX 77030 USA

The actin-myosin contractile apparatus consists of several thick filament and thin filament proteins. Specific regulatory mechanisms are involved in this highly ordered process. In this paper, we reported the identification and characterization of a novel myofibrillogenesis regulator, MR-1. The MR-1 gene was cloned from human skeletal muscle cDNA library by using a strategy that involves EST data base searching, PCR and RACE. The MR-1 gene is located on human chromosome 2.35 and encodes a 142.aa protein. Northern blot revealed that the mRNA level of MR-1 was highest in the skeletal muscle and certain level of MR-1 expression was also observed in heart, liver and kidney. Immunohistochemical assay confirmed that the MR-1 protein existed in human myocardial myofibrils. It was found by yeast two-hybrid screening and confirmed by in vitro binding assay that MR-1 could interact with sarcomeric proteins, such as myosin regulatory light chain, myomesin 1 and β-enolase. These studies suggested that MR-1 might play a regulatory role in the muscle cell and it was worth investigating further.

关键词： MR-1 gene cloning heart muscle yeast two-hybrid

Expression of MRP14 gene is frequently down-regulated in Chinese human esophageal cancer

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Cell Research 2004年第1期14卷 46-53页

作者： Jie WANG~(1*), Yan CAI~(1*), Hao XU~1, Jun ZHAO~2. Xin XU~1, Ya Ling HAN~1, Zhi Xiong XU~1, Bao Sheng CHEN~1, Hai HU~1, Min WU~1, Ming Rong WANG~(1**) ~1National Laboratory of Molecular Oncology, Cancer Institute (hospital), Chinese Academy of Medical Sciences, peking {2. of Medical School, beijing 10002., china. ~2.5} of Thoracic surgery, Cancer Institute (hospital), Chinese Academy of Medical Sciences, peking {2. of Medical School, beijing 10002., china. 1National Laboratory of Molecular Oncology Cancer Institute (Hospital) Chinese Academy of Medical Sciences Peking University of Medical School Beijing 100021 China. 1National Laboratory of Molecular Oncology Cancer Institute (Hospital) Chinese Academy of Medical Sciences Peking University of Medical School Beijing 100021 China. 1National Laboratory of Molecular Oncology Cancer Institute (Hospital) Chinese Academy of Medical Sciences Peking University of Medical School Beijing 100021 China. 2.0} of Thoracic surgery Cancer Institute (Hospital) Chinese Academy of Medical Sciences Peking University of Medical School Beijing 100021 China. 1National Laboratory of Molecular Oncology Cancer Institute (Hospital) Chinese Academy of Medical Sciences Peking University of Medical School Beijing 100021 China. 1National Laboratory of Molecular Oncology Cancer Institute (Hospital) Chinese Academy of Medical Sciences Peking University of Medical School Beijing 100021 China. 1National Laboratory of Molecular Oncology Cancer Institute (Hospital) Chinese Academy of Medical Sciences Peking University of Medical School Beijing 100021 China. 1National Laboratory of Molecular Oncology Cancer Institute (Hospital) Chinese Academy of Medical Sciences Peking University of Medical School Beijing 100021 China. 1National Laboratory of Molecular Oncology Cancer Institute (Hospital) Chinese Academy of Medical Sciences Peking University of Medical School Beijing 100021 China. 1National Laboratory of Molecular Oncology Cancer Institute (Hospital) Chinese Academy of Medical Sciences Peking University of Medical School Beijing 100021 China. 1National Laboratory of Molecular Oncology Cancer Institute (Hospital) Chinese Academy of Medical Sciences Peking University of Medical School Beijing 100021 China.

Migration inhibitory factor-related protein 14 (MRP14) is one of calcium-binding proteins, referred as S100A9. The heterodimeric molecule formed by MRP14 with its partner MRP8 (S100A8) is the major fatty acid carrier in neutrophils. The MRP8/14 complex has been also implicated in the intracellular transport of arachidonic acid and its precursors in keratinocytes. We show here the involvement of MRP14 in human esophageal cancer. In an initial study, mRNA differential display-reverse transcription polymerase chain reaction (DD-PCR) was performed with two esophageal carcinomas, one esophageal adenocarcinoma and matched normal adjacent mucosa. DD-PCR with the arbitrary primer OPA3 showed that one cDNA band was highly expressed in normal tissues, but disappeared or substantially decreased in tumor counterparts. It was later identified to be the 3’-end of migration inhibitory factor-related protein 14 (MRP14). Northern blotting, RT-PCR and Western blotting corroborated the down-regulation of MRP14 in 58/64 sq

关键词： esophagus carcinoma MRP14 S100A9.

High Jc Epitaxial Superconducting YBa_2.u_3O_(7-x) Thin Films

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Rare Metals 1990年第1期10卷 66-67页

作者： Chen Lanfeng Zhou Li Wang Chaoqun general Research Institute of Non-ferrous Metals.beijing,{2. Li Lin Zhao Bairu Zhang Yinzi Qiu Xianggang Institute of Physics,Academia Sinica,beijing.{2. Pang Shijing beijing Laboratory of Vacuum Physics,{2. Kou Yuanheng department of Physics.peking university,beijing,{2. general Research Inst of Non-ferrous Metals China

Excellent epitaxial growth of supercon-ducting YBaCuOthin films have beenrealized on(100)SrTiOand(100)ZrOsubstrates by a planar rf or DC-magnetronsputtering apparatus with UHV *** of growth and the epitaxial orientationof the film strongly depended on substratetemperature,the substrate orientation and ox-ygen partial *** films exhibitedsuperconducting onset at 92. and zero resist-ance at 90K with critical current density of

关键词： YBCO high Thin Films High Jc Epitaxial Superconducting YBa2.u3O Cu

人心肌肌钙蛋白I-C融合蛋白在大肠杆菌中的可溶性表达及纯化

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生物化学与生物物理进展 2004年第9期31卷 812-817页

作者：闫存玲徐国宾夏铁安北京大学第一医院检验科北京100034

提取人心肌组织总RNA,RT PCR扩增出人心肌肌钙蛋白I(cardiactroponinI,cTnI)和肌钙蛋白C(TnC)基因,利用人工合成的可编码 19个中性氨基酸残基为主的DNA序列(Linker),将cTnI和TnC基因连接,插入到质粒pET15b中,将鉴定正确的重组pET15b质... 详细信息

提取人心肌组织总RNA,RT PCR扩增出人心肌肌钙蛋白I(cardiactroponinI,cTnI)和肌钙蛋白C(TnC)基因,利用人工合成的可编码 19个中性氨基酸残基为主的DNA序列(Linker),将cTnI和TnC基因连接,插入到质粒pET15b中,将鉴定正确的重组pET15b质粒转化入大肠杆菌BL2.1(DE3)pLysS中,用异丙基 β D$C硫代半乳糖苷(IPTG)诱导蛋白质表达.结果在大肠杆菌中成功实现了全长cTnI C融合蛋白(cTnI linker TnC)的稳定可溶性高表达,在摇瓶中表达量可达 2.1mg/L.利用目的蛋白N端的组氨酸标签"(His tag),经一步Ni2.+ Sepharose柱亲和层析纯化,得到了聚丙烯酰胺凝胶电泳(PAGE)纯蛋白质.经初步研究,cTnI C具有较好的免疫原性和稳定性,有望成为cTnI测定系统可溯源的候选参考物质。

关键词：心肌肌钙蛋白I-C 融合蛋白表达纯化

Differential regulation of cadherin expression by osteotropic hormones and growth factors in vitro in human osteoprogenitor cells

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Acta Pharmacologica Sinica 2005年第6期26卷 705-713页

作者： Peng LIU~(2.3), Jian-hao LIN~(1,3), Bin ZHANG~1 ~1 Arthritis Clinic and Research Center, People's hospital, Medical Health Center, peking {2., beijing 100044, china ~2.department of Genetics and Developmental Biology, {2. of Connecticut Health Center, Farmington, CT 06030, USA departmentofGeneticsandDevelopmentalBiology UniversityofConnecticutHealthCenterFarmingtonCT06030USA

Aim: To examine if cadherins are expressed constitutively in human bone marrow stromal cells （hBMSC） and investigate the regulation of cadherin expression by various osteotropic hormones and local factors. Methods: Cadherin expression was examined in first passaged （secondary） hBMSC as well as in the conditionally-immortalized human osteoprogenitor cell line （hOP-7）. Using a monoclonal antibody （MoAb C-182.） to a cytoplasmic domain common to all known cadherins （pan-cadherin MoAb）, cadherins were immunolocalized in first passaged hBMSC as well as in hOP-7 cells. In addition, intense immunostaining for cadherin expression was associated with alkaline phosphatase （ALP） in nodules formed in the high density cultures of hOP-7 cells. Human E-cadherin （HECD） was specifically detected by Western blotting in extracts of untreated hBMSC using an anti-HECD MoAb 004FD. Results: Differential regulation of cadherin expression by various osteotropic hormones and local factors （parathyroid hormone, dexamethasone, estradiol, prostaglandin E2. basic fibroblast growth factor, and tumor necrosis factor-） was also observed. In addition, blocking cadherins with the MoAb C-182. increased basal ALP activity and had an additive effect on 1, 2.（OH）2.3-induced ALP activity. Conclusion: Cadherins are expressed in human osteoprogenitor cells and are involved in the osteogenic differentiation. The differential modulation of cadherin expression by osteotropic agents indicates that these agents may regulate osteoprogenitor cells through different cadherins and these cadherins may play different roles.

关键词： cadherins stromal cells cell differentiation dexamethasone prostaglandin E2 tumor necrosis factor basic fibroblast growth factor estradiol parathyroid hormone

High-and Low-Frequency Probe Tone Tympanometry in Chinese Infants

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Journal of Otology 2007年第1期2卷 42-51页

作者： SHANG Ying-ying1, NI Dao-feng1, LIU Shi-lin2.1 department of Otorhinolaryngology, peking Union Medical College {2., Chinese Academy of Medical Sciences and peking Union Medical College, beijing, china 2.department of Otorhinolaryngology, beijing Children’s {2., Capital Medical university, beijing, china department of Otorhinolaryngology Peking Union Medical College Hospital Chinese Academy of Medical Sciences and Peking Union Medical College Beijing China department of Otorhinolaryngology Beijing Children’s Hospital Capital Medical University Beijing

Objective To assess the utility of low- and high-frequency tympanometry in the diagnosis of middle ear dysfunction in Chinese infants. Methods Tympanograms were obtained with 2.6 Hz, 678 Hz and 1000 Hz probe tones from infants aged 5-2. weeks with normal auditory brainstem responses(ABRs)(15 infants, 30 ears) and with prolonged wave I latencies(17 infants, 2. ears), suggesting middle ear dysfunction, using a GSI Tympstar middle ear analyzer Version II. Results The single-peaked tympanogram was the most characteristic type in both groups and seen in 2. ears (83.3%) in the normal ABR group and in 18 ears (90%) in the delayed wave I group, respectively. The peak pressure, peak compensated static acoustic admittance and gradient of 2.6 Hz tympanometry were of no significant differences between the two groups. The 678 Hz tympanograms of admittance, susceptance and conductance demonstrated non-peak, single-, double- and tri-peaked patterns in both groups. The agreement between ABRs and 678 Hz tympanograms of admittance,susceptance and conductance were 70.0%, 58.0% and 64.0%(kappa=0.32., 0.2.4 and 0.118) respectively. For 1000 Hz probe tone, admittance, susceptance and conductance tympanograms showed single peaked patterns in 2.(93.3%), 2. (83.3%) and 2.(86.7%) of the 30 normal ears. Admittance, susceptance and conductance tympanograms using the 1000 Hz probe tone were flat in 15 (75%), 17(85%) and 13 (65%) of the ears in infants with prolonged wave I latencies. For 1000 Hz admittance, susceptance and conductance Tympanograms, the agreement between tympanometry and ABR results were 90.0%, 92.0% and 86.0% with kappa at 0.783, 0.831 and 0.690, respectively. Conclusion 1000 Hz probe tone tympanometry is a promising middle ear function test for infants of 1-6 months age, while 2.6 Hz and 678 Hz probe tones are less efficient in detecting middle ear dysfunction in infants.

关键词： ABR High-and Low-Frequency Probe Tone Tympanometry in Chinese Infants

Stromal cell can promote the proliferation of epithelial cell in prostate cancer

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Stromal cell can promote the proliferation of epithelial cel...

KAI1 inhibits HGF-induced invasion of pancreatic cancer by sphingosine kinase activity

第十三届全国泌尿外科学术会议暨第八届全球华人泌尿外科学术会议

作者： Guo Hongfeng and Na Yanqun department of Urology, peking {2. first hospital, Institute of Urology, peking {2., beijing 100034

Objective To find the effect of stromal cell on the proliferation of epithelial cell in prostatic carcinoma. Methods PC3 and PC3-AR cells were cocultured with immortal stromal cells. For experiments in vitro, we did g...

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Hepatobiliary & Pancreatic Diseases International 2011年第2期10卷 201-208页

作者： Xu Liu,Xiao-Zhong Guo,Wei-Wei Zhang,Zhuo-Zhuang Lu,Qun-Wei Zhang, Hai-Feng Duan and Li-Sheng Wang State Key Laboratory of Cancer Biology and Institute of Digestive Diseases,Xijing hospital of Digestive Disease,Fourth Military Medical university,Xi’an 710032.{2. department of Gastroenterology,Shenyang general hospital of PLA,Shenyang 110016, china department of Experimental Hematology,beijing Institute of Radiation Medicine,beijing 100850,{2. State Key Laboratory of Cancer Biology Institute of Digestive Diseases Xijing Hospital of Digestive Disease Fourth Military Medical University department of Gastroenterology Shenyang General Hospital of PLA

BACKGROUND:KAI1/CD82.has been reported to attenuate the process of metastases in a variety of tumors;however,its mechanism of action in invasion has not been fully elucidated. The present study aimed to investigate the importance of KAI1 in invasion and its correlation with activation of sphingosine kinase(SPK)in human pancreatic cancer PANC1 and Miapaca-2.cell lines. METHODS:The expression of KAI1 in PANC1 and Miapaca-2.cells,which was mediated by recombinant adenovirus(Ad-KAI1), was assessed by a flow cytometer and Western *** successful infection was established,in vitro growth curve and invasive ability in Boyden Chamber assay were *** presence of KAI1 correlating with c-Met and SPK was detected by co-immunoprecipitation and[γ-32.]ATP incorporation. RESULTS:KAI1 genes had no significant effects on the curve representing cell *** infection with the KAI1 gene,decreased invasive ability in the Boyden Chamber assay was observed in PANC1 and Miapaca-2.cells that were induced by hepatocyte growth ***-expression of KAI1 in the cells led to the deactivation of SPK and a decreased level of intracellular *** correlation was observed between c-Met and KAI1 during co-immunoprecipitation. CONCLUSION:The results of this study for the first time demonstrated a regulatory role for KAI1 in SPK activation, which leads to decreased invasive ability in disease progression of human pancreatic cancer.

关键词： adenovirus vector KAI1 gene sphingosine kinase pancreatic cancer cell lines

The role of Caveolin-1 in high glucose-induced Tissue factor activation in RAW2.4.7

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The role of Caveolin-1 in high glucose-induced Tissue factor...