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LOD score exclusion analyses for candidate disease susceptibility genes using case-parents design

Progress in Natural Science:Materials International 2006年第1期16卷 30-38页

作者： DENG Hongwen1,2,3 and GAO Guimin2,4 (1.Laboratory of Molecular and Statistical Genetics, College of Life sciences, Hunan Normal university, Changsha 410081, china 2.Osteoporosis Research Center, Creighton university medical Center, Omaha NE 68131, USA 3. department of Biomedical sciences, Creighton university, Omaha NE 68131, USA 4. Virginia Bioin format ics Institute and department of Statistics, Virginia Polytechnic Institute and State university, Blacksburg VA 24061-0315, USA) [a]Laboratory of Molecular and Statistical Genetics College of Life Sciences Human National University Changsha China [b]Osteoporosis Research Center Creighton University Medical Center Omaha NE USA [c]department of Biomedical sciences Creighton University Omaha NE USA [d]Virginia Bioinformatics Institute and department of Statistics Virginia Polytechnic Institute and State University Blacksburg VA USA

The focus of almost all the association studies of candidate genes is to test for their importance. We recently developed a LOD score approach that can be used to test against the importance of candidate genes for complex diseases and quantitative traits in random samples. As a complementary method to regular association analyses, our LOD score approach is powerful but still affected by the population admixture, though it is more conservative. To control the confounding effect of population heterogeneity, we develop here a LOD score exclusion analysis using case-parents design, the basic design of the transmission disequilibrium test (TDT) approach that is immune to population admixture. In the analysis, specific genetic effects and inheritance models at candidate genes can be analyzed and if a LOD score is ≤ - 2. 0, the locus can be excluded from having an effect larger than that specified. Simulations show that this approach has reasonable power to exclude a candidate gene having small genetic effects if it is not a disease susceptibility locus (DSL) with sample size often employed in TDT studies. Similar to association analyses with the TDT in nuclear families, our exclusion analyses are generally not affected by population admixture. The exclusion analyses may be implemented to rule out candidate genes with no or minor genetic effects as supplemental analyses for the TDT. The utility of the approach is illustrated with an application to test the importance of vitamin D receptor (VDR) gene underlying the differential risk to osteoporosis.

关键词： association studies candidate genes case-parents design disease susceptibility locus exclusion mapping LOD score.

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Role of dorsal root ganglion P2X3 receptors in electroacupuncture analgesia

Role of dorsal root ganglion P2X3 receptors in electroacupun...

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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： Zhi XIAO~1,Shan OU~(2,3) 1 department of physiology,College of basic medical sciences,Zunyi medical College,Zunyi 563003,china 2 department of Neurobiology,College of basic medical sciences,Third Military medical university,Chongqing 400038,china 3 department of Anesthesiology,General Hospital of Chengdu Military Command,Chengdu 610083,china

Background:Extracellular ATP contributes to initiation and transmission of neuropathic pain at the spinal cord and dorsal root ganglion(DRG) sites via the P2X receptors and the P2X subtype plays an important role in this *** thousands years,electroacupuncture(EA) has been clinically used to manage and alleviate chronic ***,to date,the specific analgesic mechanisms in spinal and infra-spinal levels remain unclear. Aim:The aim of this study is to investigate the effect of EA on neuropathic pain produced by chronic construction injury(CCI) mediated by P2X receptor in DRG ***:In this study,the mechanical withdrawal threshold(MWT) and thermal withdrawal latency(TWL) were investigated,and the P2X receptor protein level and the expression location in DRG were evaluated by Western blotting and *** experiment was composed by 2 series,including 6 groups,i.e.,normal group,sham CCI group,CCI group,CCI+EA group,normal+EA group and CCI+non-acupuncture point EA group(CCI+NA-EA group):Series 1,Change of pain thresholds of rats after establishment of CCI *** measurement time points involved day 0(pre-operation), day 3,day 5,day 7,day 9,day 11,day 14,day 18 and day 21 after CCI *** treatment started at day 1 after CCI operation for uninterrupted 21 *** point was"Zusanli"of contralateral operated *** 2, change of P2X receptors expression in DRG was measured by immunohistochemical and western blotting experiments on day 14 after CCI ***:The pain thresholds were decreased while P2X receptor expression in DRG was up-regulated when neuropathic pain *** the CCI group and CCI+NA-EA group,the pain thresholds were decreased dramatically after operation compared with preoperation,of which,changes on day 14 were the most *** were no significant differences in the pain thresholds among normal group,sham CCI group and normal+EA group at any time point,and the P2X receptor expressio

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Genomic research for important pathogenic bacteria in china

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Science china(Life sciences) 2009年第1期52卷 50-63页

作者： YANG RuiFu1,GUO XiaoKui2,YANG Jian3,JIANG YongQiang1,PANG Bo4,CHEN Chen5,YAO YuFeng2,QIN JinHong2 & LI QingTian2 1 Laboratory of Analytical Microbiology,State Key Laboratory of Pathogen and Biosecurity,Beijing Institute of Microbiology and Epidemiology,Beijing 100071,china 2 department of medical Microbiology and Parasitology,Institutes of medical sciences,Shanghai Jiao Tong university school of Medicine,Shanghai 200025,china 3 State Key Laboratory for Molecular Virology and Genetic engineering,Institute of Pathogen Biology,Chinese Academy of medical sciences,Beijing 100176,china 4 State Key Laboratory for Infectious Disease Prevention and Control,Institute for Communicable Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,china 5 Beijing Genomics Institute at Shenzhen,Shenzhen 518083,china 1. Laboratory of Analytical Microbiology State Key Laboratory of Pathogen and Biosecurity Beijing Institute of Microbiology and Epidemiology Beijing 100071 China 2. department of medical Microbiology and Parasitology Institutes of Medical Sciences Shanghai Jiao Tong University School of Medicine Shanghai 200025 China 3. State Key Laboratory for Molecular Virology and Genetic engineering Institute of Pathogen Biology Chinese Academy of Medical Sciences Beijing 100176 China 4. State Key Laboratory for Infectious Disease Prevention and Control Institute for Communicable Disease Control and Prevention Chinese Center for Disease Control and Prevention Beijing 102206 China 5. Beijing Genomics Institute at Shenzhen Shenzhen 518083 China

Rapid accumulation of bacterial genomic data offered an unprecedented opportunity to understand bacterial biology from a holistic view of *** can thus closely look at the way in which a pathogen is evolved,and these data has been applied to molecular epidemiology and microbial forensics,and screening of novel diagnostic,vaccine and drug *** newly developed high-throughput low-cost sequencing technologies,such as 454,Solexa and SOLiD,will promote the acquisition and application of genomic data in new research areas that we dared not imagine previously,such as the metagenomics of human gastric-intestinal tract,for better and comprehensive understanding of human health and disease.

关键词： genome pathogenic bacteria sequencing application

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Establishment of Master Cell Stock and Working Cell Bank of MDCK Lines and Selection and Evaluation of the Lines as Candidate Viral Substrates for Approval Production of Combinational Canine Attenuated-live Virus Vaccines

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Agricultural sciences in china 2002年第9期1卷 1054-1066页

作者： ZHANG De-li, LI Liu-Jin, XIA Geng-tian, HE Xu-yu, GAO Bu-xian, BAI Xiao-hong, HUANG Gao-sheng, LIU Shang-gao, [YEN Lung-fei] and FANG Fu-de( department of Immunology , school of basic medical sciences, Peking university Health Science Center , Beijing 100083 , P. R . china , Beijing Institute of Preventive Medicine, Beijing 100071, P. R . china department of Toxicology at Center for Research and Development of Animal Healthcare Products , Fourth Military medical university , Xi’an 710032 , P. R . china department of Pathology at school of Pre-clinical Education , Section of Pathology at Xi-Jing Hospital, Institute for Cancer Research at china Cancer Research Foundation , Fourth Military medical university, Xi’an 710032 , P. R . china Key Laboratory of Preventive Veterinary Medicine of State Ministry of Agriculture , College of Animal medical sciences and State Key Laboratory of Agricultural Biotechnology, College of Biological sciences , china Agricultural university, Beijing 100094 , P.R.china State Key Laboratory of medical Molecular Biology, school of basic medical sciences at the Peking Union medical Collage /Institute of basic medical sciences at the Chinese Academy of medical sciences , Beijing 100005 , P. R . china ) department of Immunology School ofBasic Medical Sciences Peking University Health Science Center Beijing 100083 P.R. China Beijing Institute of Preventive Medicine Beijing 100071 P. R. China department of Toxicology at Center for Research and Devel department of Toxicology at Center for Research and Development of Animal Healthcare Products Fourth Military Medical University Xi'an 710032 P. R. China Beijing Institute of Preventive Medicine Beijing 100071 P. R. China department of Pathology at school of Pre-clinical Education Section of Pathology at Xi-Jing Hospital Institute for Cancer Research at China Cancer Research Foundation Fourth Military Medical University Xi'an 710032 P. R. China Key Laboratory of Preventive Veterinary Medicine of State Ministry of Agriculture College of Animal Medical Sciences and State Key Laboratory of Agricultural Biotechnology College of Biological Sciences China Agricultural UniversityBeijing 100094 P. State Key Laboratory of medical Molecular Biology School of Basic Medical Sciences at the Peking Union Medical Colloge /Institute of Basic Medical Sciences at the Chinese Academy of Medical Sciences Beijing 100005 P. R . China

Under the prerequisite that the incidence of cancer or tumor in negative-control nude mice inoculated subcutaneously with primary feline or canine kidney cell cultures purified in vitro at passage 3 was 0 (0/22) and 0 (0/10), respectively. The incidence of the progressively-growing malignant tumor(MT) in positive-control nude mice inoculated subcutaneously with Hela cell cultures of KB, X, or NM20/X strain was 10/ 10, 25/25 and 5/51, respectively. The results showed that the incidence of tumor in nude mice with di-and hy-perploid YB strain of MDCK cell during 17 - 23 passages, with hyper- and hypoploid KA strain of MDCK cell during 6-8 passages, with hypoploid WB strain of MDCK cell on passage 6, with hyper-and hypoploid H strain of MDCK cell during 8-24 passages was 2/24, 6/10, 5/10 and 10/15, respectively. The chromosomal analysis results showed that the ratio of difference in the rate of modal chromosome number between high(mcs + n) and lowest (mcs)passages was not more than 5 - 15% and the structure aberrations was generally 0 -3 %. These results proved that the genetic characteristics of chromosomal number of cell lines determines their tumorigenicity, but it is species-specific. MDCK line has tumorigenicity no matter what its chromosome kary-otype is, at least it has very low tumorigenicity even when its modal chromosome number is hypoploid. It is thus evident that MDCK cell of WB or H strain can be approved as substrate for the preparation of attenuated viral vaccines, but MDCK cell of YB or KA strain can not be approved as substrate for the preparation of attenuated viral vaccines.

关键词： Cell line MDCK Master cell stock Working cell bank Karyotypes Carcinogenesis Tu-morigenicity

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Effect of PDS and lycopene on learning and memory of senile mice cumulating with lactic acid in atlanto-axial joint

Effect of PDS and lycopene on learning and memory of senile ...

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2008 International Conference on Ginseng

作者： FAN Wen-jing ZHU Tong-tong GUO Ya-xiong LIN Gui-miao ZHAO Li-juan department of Pathophysiology school of basic medicine sciences jilin university changchun 130021

＜正＞Objective To compare PDS with lycopene in the function of the improvement of learning and memory abilities by cumulating lactic acid for long time between atlas dentata vertebrae in senile *** 30% lactic acid 30μL one time every week was injected into the place between the first and the second cervical vertebrae,three times was *** changes of learning and memory in mice were observed with water-maze *** activities of acetylcholine esterase（ChE）, and the content of acetylcholine,（ACh）in encephalon were tested,Immunohistochemical staining withβ-amyloid protein in mouse brain,including the cortex and hippocampus was *** The model mice had longer swimming full time and more error times（p＜0.05）in water maze test compared with control *** could significantly shortened the latency period and wrong times in water-maze test（P＜0.01）,but lycopene’mice showing was not *** with model group,the activities of ChE increased（P＜0.01）,and the content of ACh decreased in the PDS and lycopene group（P＜0.01）,and in the two therapy groups,β-amyloid protein in mouse brain of cortex and hippocampus was less than the model ***:Our findings indicate that PDS and lycopene has improved the learning and memory abilities,as well as its ability to ameliorate the metabolism in senile mice encephalon, which may be the mechanism of its cerebral protective effect.

关键词： PDS lycopene learning and memory atlanto-axial joint β-amyloid

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Overexpression of heat-shock protein 20 in rat heart myogenic cells confers protection against simulated ischemia/reperfusion injury

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Acta Pharmacologica Sinica 2005年第9期26卷 1076-1080页

作者： Yan-hui ZHU Xian WANG~2 department of physiology and Pathophysiology,basic medical College,Key Laboratory of Molecular Cardiovascular Biology,Ministry of Education,Peking university,Beijing 100083,china departmentofphysiologyandPathophysiology BasicMedicalCollegeKeyLaboratoryofMolecularCardiovascularBiologyMin

Aim:To explore whether overexpression of the small heat shock protein HSP20 inrat cardiomyocytes protects against simulated ischemia/reperfusion(SI/R)***:Recombinant adenovirus expressing HSP20 was used to infect rat H9c2cardiomyocytes at high efficiency,as assessed by green fluorescent protein.H9c2cells were subjected to SI/R stress;survival was estimated through assessment oflactate dehydrogenase and cell apoptosis through caspase-3 ***:Overexpression of HSP20 decreased lactate dehydrogenase release by 21.5% andcaspase-3 activity by 58.8%.Pretreatment with the protein kinase C inhibitor Ro-31-8220(0.1 μmol/L)for 30 min before SI/R canceled the protective effect of *** selective mitochondrial K+awP channel inhibitor 5-hydroxydecanoate(100 μmol/L)had a similar ***,the non-selective K~+ATPchannel inhibitorglibenclamide(100 μmol/L)had no significant ***:These dataindicate that the protective effect of HSP20 in vitro is primarily due to reducednecrotic and apoptotic death of cardiomyocytes,possibly via the protein kinaseC/mitochondrial K~+ATP pathway.

关键词： reperfusion injury adenovirus apoptosis necrosis heat-shock proteins

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Temporal Integration of Sound Intensity Determines the Firing Threshold of an Inferior Collicular Neuron in Mice

Temporal Integration of Sound Intensity Determines the Firin...

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第十一次中国生物物理学术大会暨第九届全国会员代表大会

作者： Xiao Wang,Weiguo Yang,Xiaodong Tan,Zhongju Xiao department of physiology,school of basic medical sciences,Southern medical university,Guangzhou 510515

＜正＞Neuron discharging is thought to be triggered by a fixed threshold *** auditory system studies,however,what is the neuronal firing threshold has been debated not only in

关键词： first spike latency envelope of tone burst neuron auditory system

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Effect of transforming growth factor-beta on activity of connective tissue growth factor gene promoter in mouse NIH/3T3 fibroblasts

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Acta Pharmacologica Sinica 2004年第4期25卷 87-91页

作者： Qing ZHAO, Nan CHEN2, Wei-ming WANG, Jian LU3,Bing-bing DAI3 department of Nephrology, Ruijin Hospital 3department of Biochemistry, Shanghai Second medical university, Shanghai 200025, china department of Nephrology Ruijin Hospital

AIM: To investigate the regulatory mechanism of transforming growth factor-betaon activity of connective tissue growth factor promoter in mouse NIH/3T3 fibroblasts. METHODS: The regulation fragment of the 5' flanking region of the human CTGF gene was linked to pGL3-basic vector, a firefly luciferase reporter construct without promoter. The recombinant plasmid pCTGF-luc was transiently transfected to NIH/3T3 fibroblasts. The activity of CTGF promoter after treatment of TGF-β1 and MAPK pathway inhibitors were assayed with luciferase reporter gene assay system. RESULTS: TGF-β1-induced increase of CTGF promoter activity was concentration-dependent, with a plateau at 5 μg/L by 2.67-fold vs control (P2 h and maintained to 24 h by 2.76- and 2.20-fold vs control, respectively (P203580 (10 μmol/L), the p38 MAP kinaseinhibitor, decreased basal and TGF-β1-induced activation of CTGF promoter. However, inhibition of c-Jun-N-terminal kinase/stress-activated protein kinase by SP600125 (20 μmol/L) was without effect. CONCLUSION: TGF-β1stimulated the transcriptional activity of CTGF gene promoter in NIH/3T3 fibroblasts in a dose- and time-dependent manner. MAPK pathway may play a role in the regulation of TGF-β1-induced CTGF expression.

关键词： transforming growth factor-beta connective tissue growth factor promoter mitogen-activated protein kinases signal transduction

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Human endothelial senescence induced by IL-1α in vitro

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Progress in Natural Science:Materials International 2004年第3期 26-30+97页

作者： YAO Aiyu 1,2 , ZHOU Jianjun 1, LIU Yabing 1, FENG Meifu 1* and ZHOU Rouli 2*(1. State Key Laboratory of Biomembrane and Membrane Biotechnology, Institute of Zoology, Chinese Academy of sciences, Beijing 100080, china 2. department of Cell Biology and medical Genetics, school of basic medical sciences, Health Science Center, Peking university, Beijing 100083, china) StateKeyLaboratoryofBiomembraneandMembraneBiotechnology InstituteofZoologyChineseAcademyofSciencesBeijing100080China departmentofCellBiologyandmedicalGenetics SchoolofBasicMedicalSciencesHealthScienceCenterPekingUniversityBeijing100083China

Interleukin 1 (IL 1) is an important proinflammatory cytokine that causes pleiotropic effects. Vascular endothelial cells stimulated by IL 1α can lead to the inflammatory response. Reactive oxygen species (ROS) are also generated at the site of inflammation and serve as an important factor against foreign invader. Here we report that long term stimulation of human vein endothelial cells with IL 1α can accelerate their senescence associated with β galactosidase activity. The flow cytometric analyses showed that most of the induced cells entered G 0 G 1 phase. DNA damage was more severe in senescent cells by comet assay. The induced cells by IL 1α had higher levels of ROS and malonyldialdehyde (MDA), lower activity of antioxidant enzymes and lower capacity of total antioxidant systems than control, which led to cell damage and cell degeneration, that is to say, which contributed to cellular senescence. Our results gave a direct proof to a new hypothesis—“the inflammation hypothesis of aging” on cellular level, and also provided a basis for the study on anti aging and aging related diseases.

关键词： IL 1α endothelial cell senescence ROS antioxidant systems.

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Cloning of Chicken Anemia Virus vp3 Gene and Apoptosis Inductive Effect of vp3 Gene In Vitro

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Journal of Huazhong university of Science and Technology(medical sciences) 2003年第4期23卷 329-331,334页

作者：孙军王宇哲宗义强屈伸 department of Biochemistry and Molecular Biology

Using PCR technique, the vp3 gene of chicken anemia virus (CAV) was cloned into the eukaryotic expression vector pcDNA3 to construct a recombinant pcDNA vp3. Restriction enzyme digestion and sequencing analysis revealed that CAV vp3 gene was correctly inserted into the blank vector pcDNA3. After LipofectAMINE TM mediated transfection in vitro with pcDNA vp3 and pcDNA3 respectively, the total mRNA was extracted from liver carcinoma cell lines HepG2 and diploid cell line L 02, and RT PCR was performed afterward. The results of RT PCR suggested that vp3 gene was expressed in these two cell lines. At the same time, using in situ apoptotic detection assay, TUNEL kits, the apoptotic cells were found in pcDNA vp3 transfected HepG2, but not in mock transfected cell lines. VP3 could induce cell death by apoptosis in cancer cell lines, but not in diploid cell lines. All the results indicated that CAV vp3 gene, a potential therapeutic agents, has the potential of being used for cancer treatment.

关键词： chicken anemia virus gene cloning apoptosis vp3 gene

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