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Adeno-associated virus-mediated bone morphogenetic protein-7 gene transfer induces C2C12 cell differentiation into osteoblast lineage cells

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Acta Pharmacologica Sinica 2005年第8期26卷 963-968页

作者： Min YANG~2, Qing-jun MA~(2,4., Geng-ting DANG~2, Kang-tao MA~3. Ping CHEN~3. Chun-yan ZHOU~(3.4. ~2department of Orthopedics,{4. third hospital,{3., china ~3.0} of Biochemistry and Molecular Biology, School of Basic Medical Sciences,{4. Health Science Center,{3., china department of Orthopedics Peking University Third Hospital Beijing China department of Biochemistry and Molecular Biology School of Basic Medical Sciences Peking University Health Science Center Beijing China

Aim: To investigate the effects of bone morphogenetic protein-7 (BMP7)-expressing recombinant adeno-associated virus (AAV) vector on the differentiation of C2C12 cells. Methods: AAV-BMP7 was packaged by infecting the stable cell clone BHK-21 (integrated with recombinant AAV vector plasmid pSNAV-BMP7) with recombinant herpes simplex virus type 1, which expresses AAV-2 Rep and Cap and possesses AAV packaging functions. Following infection with AAV-BMP7 at multiplicities of infection of lx 10～5 vector genomes per cell and subse-quent culture, C2C12 cells were assessed qualitatively for BMP7 production, alka-line phosphatase activity, osteocalcin production and Cbfal and MyoD expression. Results: C2C12 cells transduced with AAV-BMP7 could produce BMP7 protein until d 28. Alkaline phosphatase in the cultured C2C12 cell lysate was elevated. Secreted osteocalcin in the culture medium was detectable at d 12 and Cbfal mRNA expression level was upregulated, coinciding with downregulation of MyoD in a temporal manner. Conclusion: The present in vitro study demonstrated that AAV-BMP7 could infect and efficiently convert C2C12 cells from myoblasts into osteoblast lineage cells.

关键词： C2C12 adeno-associated virus bone morphogenetic protein-7 gene therapy

AICAR inhibits proliferation and induced S-phase arrest,and promotes apoptosis in CaSki cells

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Acta Pharmacologica Sinica 2007年第12期28卷 1984-1990页

作者： Tong-ju GUAN~2,Feng-jin QIN~3.Jian-hai DU~4.Li Geng~3.You-yi ZHANG~(4.5),Min LI~(2,5)~2 Institute of Pathology,The School of Basic Medicine,Lanzhou university,Lanzhou 73.000,{4. ~3.department of Gynaecology andObstetrics,{3. university third hospital,beijing 100083,{4. ~4.Institute of Vascular Medicine,{3. university third hospital andKey Laboratory of Molecular Cardiovascular Sciences Ministry of Education,beijing 100083,{4. Institute of Pathology The School of Basic Medicine Lanzhou University Lanzhou China department of Gynaecology and Obstetrics Peking University Third Hospital Beijing China Institute of Vascular Medicine Peking University Third Hospital and Key Laboratory of Molecular Cardiovascular Sciences Ministry of Education Beijing China

Aim:The aim of the present study was to determine the effect of 5-aminoimidazole-4.carboxamide-ribonucleoside （AICAR） on proliferation,cell cycle,and apoptosisin the human epithelial cervical cancer cell line CaSki ***:Cell countand 3.（4.5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay were usedto determine cell proliferation and *** 3.258 staining was con-ducted to distinguish the apoptotic *** cycle and Annexin-V/propidiumiodide staining were analyzed by fluorescence-activated cell sorting （FACS）.AWestern blot assay was used to evaluate the expression of AKT （also known asprotein kinase B）,mammalian target of rapamycin （mTOR）,p53.and extracellularsignal-regulated kinase （ERK）.Results:AICAR （500 μmol/L） significantly inhibi-ted the proliferation of CaSki cells treated for 24.4.,and 72 h as determined by *** cells at the G1and G2phases were dramatically decreased while cells atthe S phase were increased in response to AICAR treatment for 24.4.,and 72 *** MTT assay showed less viable cells and Hoechst fluorescent staining showedmore apoptotic cells upon AICAR *** results of the Annexin-Vstaining demonstrated a time-dependent increase of apoptosis in cells treatedwith AICAR for 24.3.,and 4. ***,AICAR activated caspase-3.in atime-dependent *** was also found that AICAR inhibited the phosphory-lation of AKT and mTOR,which are important kinases regulating cell growth *** stimulation obviously increased the expression of the tumorsuppressor p53.and the phosphorylation of ***:AICAR inhibitedproliferation and induced S phase arrest and promoted apoptosis in CaSki cells,which might be mediated by the downregulation of the AKT/mTOR pathway andthe upregulation of the p53.ERK pathway.

关键词： AICAR proliferation apoptosis CaSki cells

Probability parametric imaging with DCE-MRI data to identify prostate cancer in the peripheral zone

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Progress in Natural Science:Materials International 2006年第9期16卷 948-953页

作者： HUANG Xinrui, WANG Qiang, XU Yufeng, WANG Xiaoying and BAO Shanglian ( The Key Laboratory of Medical Physics and Engineering, peking university, beijing 100871, china The Medical School of peking university,{4., china 3. The First hospital of peking university, beijing 10003., china) The Key Laboratory of Medical Physics and Engineering Peking University Beijing 100871 China The Medical School of peking university Beijing 100083 China The First hospital of peking university Beijing 100034 China

To quantitatively analyze prostate dynamic contrast-enhanced magnetic resonance images (DCE-MRI), a retrospective study was carried out to construct a mathematical model to predict whether a lesion is cancerous or not. With known diagnosis of 52 re-gions-of-interest (10 malignant, 26 benign and 16 normal) in peripheral zone (PZ) of prostate from 18 patients undergoing needle biopsy and 8 normal volunteers, we combined feature parameters (k1, k2 and tp) from the linear-slope model with logistic regression to determine the statistical probability model. Prostate cancerous probability maps were derived using the model on each subject to examine the correlation between the model-based prediction and the known diagnosis. Compared the model prediction results of 52 regions-of-interest with the histopathologic results, the total agreement rate was 90.4.% ; the agreement rate was 60 % for the malignant and 97. 6 % for the nonmalignant. The diagnostic results only referencing the probability maps by two radiologists showed a high agreement in tissue characteristics and lesion location with the combining results from other diagnostic techniques such as PSA (prostate specific antigen) level and MRS (magnetic resonance spectroscopy) .

关键词： time-intensity curve prostate DCE-MRI logistic regression probability parametric image.

Distinguishing malignant from normal stomach tissues and its in vivo, in situ measurement in operating process using FTIR Fiber-Optic techniques

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Science china Chemistry 2005年第2期48卷 167-175页

作者： XU Yizhuang1, YANG Limin1, XU Zhi3. ZHAO Ying2, LING Xiaofeng3. LI Qingbo1, WANG Jiansheng4. ZHANG Nengwei3. ZHANG Yuanfu1 & WU Jinguang1 1. The State Key Laboratory of Rare Earth Materials and Applications, College of Chemistry and Molecular Engineering, peking university, {4. 100871, china 2. Institute of Chemistry, Chinese Academy of Sciences, {4. 100080, china 3. department of Surgery, third hospital, peking university, beijing 100083, china 4. department of Oncology Surgery, the First hospital of Xi’an Jiaotong university, Xi’an 710061, china Correspondence should be addressed to Wu Jinguang (email: wujg@***) The State Key Laboratory of Rare Earth Materials and Applications College of Chemistry and Molecular Engineering Peking University Beijing China department of Surgery Third Hospital Peking University Beijing China Institute of Chemistry Chinese Academy of Sciences Beijing China department of Oncology Surgery the First Hospital of Xi’an Jiaotong University Xi’an China

Gastric tissue samples were studied using mid-IR fiber-optic attenuated total reflectance (ATR) spectroscopy. FTIR spectra of 90 tissue samples from 4. patients, including 3. normal and 58 malignant tissue samples, were chosen as examples. Malignancy was usually characterized by the absence of CH and C=O bands, a weak amide II band near 154. cm-1, a shift of the amide I band to lower wave number, a decrease in the ～14.0 cm-1 peak to less than the～14.0 cm-1 peak. Subtraction spectra indicate that the amide I and amide II bands of normal and malignant tissues have larger differences in peak positions and relative intensities. The statistical analysis results confirm this conclusion. The results indicate that FTIR fiber optic techniques provide important information about cancerous tissue of the stomach, which can be used to differentiate the malignant tissue from the normal tissue. Based on the above results we successfully realize the detection of the tumor tissues of digestive tract in vivo and in situ. And the results of detection cancer near operating room and in vivo and in situ in the operating room are consistent with the conclusions for the samples stored in liquid N2, which is the basis for the clinical application.

关键词： FTIR, Fiber-Optics, stomach tissues, cancer, diagnosis.

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Sodium hydrosulfide alleviated pulmonary vascular structural remodeling induced by high pulmonary blood flow in rats

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Acta Pharmacologica Sinica 2006年第8期27卷 971-980页

作者： Xiao-hui LI~2 Jun-bao DU~(2,6) Ding-fang BU~3.Xiu-ying TANG~4.Chao-shu TANG~5 ~1department of Pediatrics,peking university First {3.,Key Laboratory of Molecular Cardiovascular Diseases,Ministry of Education ~3.Laboratory Center ~4.aboratory of Electron Microscopy,peking university First {3.,beijing 10003.,china ~5department of Physiology,peking university Health Science Center,beijing {13.,china department of Pediatrics Peking University First Hospital Key Laboratory of Molecular Cardiovascular Diseases Ministry of Education Beijing China Laboratory Center Peking University First Hospital Beijing China Laboratory of Electron Microscopy Peking University First Hospital Beijing China department of Physiology Peking University Health Science Center Beijing China

Aim:To explore the possible role of endogenous hydrogen sulfide （H2S）,a novelgasotransmitter,in the pathogenesis of pulmonary vascular structural remodeling（PVSR） induced by high pulmonary blood ***:Thirty-two Sprague-Dawley male rats were randomly divided into sham,shunt,sham+NaHS （a H2Sdonor） and shunt+NaHS *** in shunt and shunt+NaHS groups under-went an abdominal aorta-inferior vena cava shunt,and rats in shunt+NaHS andsham+NaHS groups were intraperitoneally injected with *** was inves-tigated using optical microscope and transmission electron *** H2S was evaluated by sulfide-sensitive *** oxide synthase（NOS）,heme oxygenase （HO-1）,proliferative cell nuclear antigen （PCNA） andextracellular signal-regulated kinase （ERK） activation were analyzed by ***:After 11 weeks of shunting,PVSR developed with a decrease inlung tissue H2S production and an increase in nitric oxide （NO）.However,lungtissue carbon monoxide （CO） did not *** the treatment with NaHS for11 weeks,H2S donor ameliorated PVSR and downregulated PCNA expression andERK activation with an increase in lung tissue CO production and HO-1 proteinexpression but a decrease in NO production,NOS activity and eNOS proteinexpression in shunted ***:H2S exerted a regulatory effect on PVSRinduced by high pulmonary blood ***,H2S down-regulated theERK/MAPK signal pathway,inhibited the NO/NOS pathway and enhanced theCO/HO pathway in rats with high pulmonary blood flow.

关键词： hydrogen sulfide pulmonary hypertension structure remodeling nitric oxide/nitric oxide synthase carbon monoxide/heme oxygenase

Rat adipose-derived stromal cells expressing BMP4.induce ectopic bone formation in vitro and in vivo

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Acta Pharmacologica Sinica 2006年第12期27卷 1608-1615页

作者： Lin LIN~2 Xin FU~2 Xin ZHANG~2 Lian-xu CHEN~2 Ji-ying ZHANG~2 Chang-long YU~(2,4.Kang-tao MA~3.Chun-yan ZHOU~(3.4.2 Institute of Sports Medicine,peking university{4.,{3. 100083,china 3.department of Biochemistry and Molecular Biology,peking university School of Basic Medical Science,{3. 100083,china Institute of Sports Medicine Peking University Third Hospital Beijing China department of Biochemistry and Molecular Biology Peking University School of Basic Medical Science Beijing China

Aim:Bone morphogenetic protein 4.BMP4.is one of the main local contributingfactors in callus formation in the early phase of fracture ***-derivedstromal cells(ADSC)are multipotent *** present study was conducted toinvestigate the osteogenic potential of ADSC when exposed to adenovirus con-taining BMP4.cDNA(Ad-BMP4..Methods:ADSC were harvested from Sprague-Dawley *** exposure to Ad-BMP4.ADSC were assessed by alkaline phos-phatase activity(ALP)assay,RT-PCR and yon Kossa ***4.expressionwas assessed by RT-PCR,immunofluorescence and Western blot analysis,ADSCtransduced with Ad-BMP4.were directly injected into the hind limb muscles ofathyrnic *** Ad-EGFP(enhanced green fluorescence protein)served *** animals were examined by X-ray film and histological ***:The expression of BMP4.was confirmed at both mRNA and protein *** expression of the osteoblastic gene,ALP activity and yon Kossa stainingconfirmed that ADSC transduced with Ad-BMP4.underwent rapid and markedosteoblast differentiation,whereas ADSC transduced with Ad-EGFP and cells leftalone displayed no osteogenic differentiation.X-ray and histological examinationconfirmed new bone formation in athymic mice transplanted with ADSC trans-duced with ***:Our data demonstrated successful osteo-genic differentiation of ADSC transduced with Ad-BMP4.in vitro and in *** may be an ideal source of mesenchyme lineage stem cells for gene therapyand tissue engineering.

关键词： adipose-derived stromal cells BMP4 gene therapy bone formation

Progress and review of the studies on the end-Triassic mass extinction event

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Science china Earth Sciences 2005年第12期48卷 2049-2060页

作者： DENG Shenghui1,2, LU Yuanzheng1,2,3.& XU Daoyi4.1. Research Institute of Petroleum Exploration and Development, beijing 100083, china 2. Key Laboratory of Oil and Gas Reservoir of Petrochina, beijing 100083, china 3. china university of Geosciences, beijing 100083, china 4. Institute of Geology, china Earthquake Administration, beijing 100029, china Research Institute of Petroleum Exploration and Development Beijing China Key Laboratory of Oil and Gas Reservoir of Petrochina Beijing China china university of Geosciences Beijing China Institute of Geology China Earthquake Administration Beijing China

The mass extinction at the end-Triassic is one of the five biggest in the Phanerozoic. However,it is the least well understood among these five events, and only till last decade it became a great academic interesting subject to geologists. The evidences for this event come obviously from bivalves, brachiopods, ammonites, corals, radiolaria, ostracods and foraminifera of marine habitats, and plants and tetrapods of terrestrial realm. By contrast, for some of other groups, such as marine gastropods and marine vertebrates, no mass extinction has been recog-nized yet. The extinction event is strongly marked at specific level but shown a complicated situa-tion at generic and family levels. Dramatic changing of the environment, such as the temperature raise due to the greenhouse effect, the marine anoxic habitats caused by a sudden transgression after the regression at the end of Triassic, has been claimed to be the main cause of the extinction. Many hypotheses have been suggested to demonstrate the mechanisms of the environment changing, among which two popular ones are the bolide impact and volcanic eruption. The Triassic-Jurassic (Tr-J) boundary mass extinction event is still poorly understood because no enough data have been obtained from the Tr-J boundary successional sections of both marine and terrestrial sediments, and most of these studies were regionally restricted. The basic aspects of the event and its associated environmental conditions remain poorly characterized and the causal mechanism or mechanisms are equivocal. Some authors even doubt its occurrence. china has many successionally developed terrestrial and marine Tr-J sections. Detailed studies of these sections may be important and significant for well understanding of the event.

关键词： progress, review, end-Triassic, mass extinction, mechanism, Tr-J boundary.

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Population pharmacokinetics of valproate in Chinese children with epilepsy

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Acta Pharmacologica Sinica 2007年第10期28卷 1677-1684页

作者： De-chun JIANG~(2,3.,Li WANG~4.Yu-qin WANG~(2,6),Lin LI~3.Wei LU~5,Xiang-rong BAI~2~2 department of Pharmacy,Xuan-wu hospital of Capital Medical {4.,{3. 100053.china ~4.department of Pediatrics,peking {4.First hospital,{3. 10003.,china ~3.department of Pharmacology,Xuan-wu hospital of Capital Medical {4.,Education Ministry KeyLaboratory for Neurodegenerative Diseases,{3. 100053.china ~5 School of Pharmaceutical Science,peking {4.,{3. 100083,china department of Pharmacy Xuan-wu Hospital of Capital Medical University Beijing China department of Pharmacology Xuan-wu Hospital of Capital Medical University Education Ministry Key Laboratory for Neurodegenerative Diseases Beijing China department of Pediatrics Peking University First Hospital Beijing China School of Pharmaceutical Science Peking University Beijing China

Aim:The aim of the present study is to establish a population pharmacokinetic（PPK） model of valproate （VPA） in Chinese epileptic children to promote thereasonable use of anti-epileptic ***:Sparse data of VPA serumconcentrations from 4.7 epileptic children were *** patients weredivided into 2 groups:the PPK model group （n=3.7） and the PPK valid group（n=100）.The PPK parameter values of VPA were calculated by NONMEM soft-ware using the data of the PPK model group.A basic model and a final modelwere set *** validate the 2 models,the concentrations of PPK valid groupwere predicted by each model,*** mean prediction error （MPE）,mean squared prediction error （MSPE）,root mean squared prediction error（RMSPE）,weight residues （WRES）,and the 95% confidence intervals （95%CI） were also ***,the values between the 2 models were ***:The PPK of VPA was determined by a 1-compartment model with afirst-order absorption *** basic model was:Ka=3.09 (h-1),V/F=20.4.L）,CL/F=0.296 （L/h）.The final model was:Ka=0.251+2.24.（1-HS）(h-1),V/F=2.88+0.157·WT （L）,CL/F=0.1060.98·CO+0.0157·AGE （L/h）.For thebasic model,the MPE,MSPE,RMSPE,WRES,and the 95% CI were-23.53.-3..3.,-16.70）,3.28.96 （2872.72,4.85.20）,3..62 （3..3.,4..90）,and-0.06 （-0.14.0.02）,*** the final model,the MPE,MSPE,RMSPE,WRES,and the95% CI were-1.16 （-4.85,2.53.,1002.83.（1050.64.114.0}61）,23.04.（21.12,24.96）,and0.08 （-0.04.0.20）,*** final model was more optimal than the ***:The PPK model of VPA in Chinese epileptic children wassuccessfully *** will be valuable to facilitate individualized dosageregimens.

关键词： population pharmacokinetics valproic acid epilepsy children Bayesian method NONMEM software individualized dosage regimens

Taurine inhibits ischemia/reperfusion-induced compartment syndrome in rabbits

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Acta Pharmacologica Sinica 2005年第7期26卷 821-827页

作者： Ji-xian WANG~(2,3. Yan LI~2 Li-ke ZHANG~4.Jing ZHAO~5 Yong-zheng PANG~(2,3. Chao-shu TANG~(3.5) Jing ZHANG~(2,3.6) 1 Institute of Cardiovascular Disease Research,The First {4. of peking university,beijing 10003.,{3. 3.Key Laboratory of Molecular Cardiovascular Science,Ministry of Education,beijing 100083,{3. 4.department of Pathophysiology,Capital Institute of Medicine,beijing 100054.{3. 5 department of Physiology Health Center of Medical Sciences,peking university,beijing 100083,{3. Institute of Cardiovascular Disease Research The First Hospital of Peking University Beijing China Key Laboratory of Molecular Cardiovascular Science Ministry of Education Beijing China department of Pathophysiology Capital Institute of Medicine Beijing China department of Physiology Health Center of Medical Sciences Peking University Beijing China

Aim:To investigate effects of taurine on ischemia/reperfusion(I/R)-induced com-partment syndrome in rabbit hind ***:Rabbits underwent femoralartery occlusion after ligation of branches from terminal aorta to femoral *** a 7-h ischemia,reperfusion was established with the use of heparinizedpolyethylene *** received taurine(1 g/kg)or normal saline(control)by iv infusion 10 min before shunt *** reperfusion,anterior com-partment pressure(ACP)was monitored continuously in the left lower *** muscle triphenyltetrazolium chloride(TTC)level,taurine contentand myeloperoxidase activity were *** stress was induced in thein vitro gastrocnemius muscle slices by free radical generating systems(FRGS),and the malondialdehyde content was measured in presence or absence of ***:After 7 h of ischemia,none of the parameters that we measured weredifferent from those before ischemia,except that TTC reduction decreased by80%.In the control group,after 2 h of reperfusion,ACP increased 4.5-fold,andgastrocnemius muscle taurine content was reduced by 3.%.In taurine-treatedanimals,at 2 h reperfusion,the mean arterial blood pressure and heart rate wereincreased,by 6% and 10%.ACP decreased by 3.%,muscle edema decreased by16%,TTC reduction increased by 150%,and lactate dehydrogenase decreased by3.% compared to control *** and muscle taurine content increased by70% and 88%,*** the taurine-treated group,at 2 h repeffusion,plasmamalondialdehyde and conjugated diene content were decreased by 3.% and 23.,respectively,and muscle malondialdehyde and conjugated diene content decreasedby 22% and 3.%,respectively compared to the control *** 2 h reperfusion,myeloperoxidase activity was increased 3.5-fold in control *** the in vitrostudy,taurine decreased malondialdehyde content in muscle slices incubatedwith hypochlorous acid in a dose-dependent manner,but there was no changewhen incubated

关键词： taurine compartment syndromes reperfu sion injury