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Antisense oligonucleotides-induced local blockade of T-bet expression leads to airway inflammation in rats

Acta Pharmacologica Sinica 2006年第5期27卷 561-567页

作者： Gang WANG~2 Chun-tao LIU~(3,4) Zeng-li WANG~3 Li-li JIANG~5 Cun-liang YAN~6 Feng-min LUO~3 ~2department of Integrated Traditional Chinese and western medicine.west china hospital.sichuan university,chengdu 610041,china.~3department of Respiratory medicine,west china hospital.sichuan university.chengdu 610041.china ~5department of pathology,west china hospital.sichuan university,chengdu 610041,china:~6department of laboratory medicine.shenzhen hospital,peking university.shenzhen 518036,china department of Integrated Traditional Chinese and western medicine West China Hospital Sichuan University Chengdu China department of Respiratory medicine West China Hospital Sichuan University Chengdu China department of pathology West China Hospital Sichuan University Chengdu China department of laboratory medicine Shenzhen Hospital Peking University Shenzhen China

Aim:To explore whether local blockade of T-box expressed in T cells (T-bet)expression in the lungs could lead to airway ***:Twenty-fourrats were randomly divided into 4 groups:saline group,ovalbumin (OVA)-sensi-tized group,nonsense group,and the antisense *** OVA-sensitized ratswere sensitized and challenged with OVA,and the rats in the nonsense andantisense groups were subjected to an aerosol delivery of the nonsense andantisense oligonucleotides (AS-ODN) of T-bet (0.1%,w/v).The levels of inter-feron-γ(IFN-γ),interleukin-4(IL-4),and IL-5 in the bronchoalveolar lavage fluid(BALF) were detected by ELISA,and the mRNA and the protein expression ofT-bet and GATA-3 genes were examined by in situ hybridization and western blotanalysis,***:The expression of T-bet mRNA and protein in thelungs of the rats in the antisense group were inhibited *** lungs ofthe rats in the antisense and OVA-sensitized groups showed eosinophil and lym-phocyte inflammatory infiltration,and eosinophilia located predominantly aroundthe *** number of GATA-3 mRNA-positive cells and the level of GATA-3protein in the lungs of the rats in the antisense and the OVA-sensitized groupssignificantly *** level of IL-4 and IL-5 in the BALF in the antisenseand OVA-sensitized groups were elevated,but the level of IFN-γ ***:Antisense ODN-induced local blockade of T-bet expres-sion leads to airway inflammation with a selective alteration in patterns of cytokineexpression and recruitment of eosinophil cells similar to that in the OVA-sensitizedanimals.

关键词： airway inflammation antisense oligonucleotides GATA3 transcription factor interferon type Ⅱ interleukin-4 interleukin-5 Tbox expressed in T cells

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A Novel Missense Mutation(L296Q)in Cholesteryl Ester Transfer Protein Gene Related to Coronary Heart Disease

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Acta Biochimica et Biophysica Sinica 2004年第1期36卷 33-36页

作者： Ke-Qin ZHENG, Si-Zhong ZHANG, Li ZHANG, De-Jia HUANG, Lin-Chuan LIAO, and Yi-Ping HOU ( department of Medical Genetics, west china hospital Division of Human Morbid Genomics, Key laboratory ofBiotherapy of Human Diseases of Ministry of Education, sichuan university, chengdu 610041, china department of Cardiology, west china hospital, sichuan university, chengdu 610041, china School of Basic and Forensic medicine, sichuan university, chengdu 610041, china) department of Medical Genetics West China Hospital Division of Human Morbid Genomics Key Laboratory of Biotherapy of Human Diseases of Ministry of Education Sichuan University Chengdu 610041 China department of Cardiology West China Hospital Sichuan University Chengdu 610041 China School of Basic and Forensic medicine Sichuan University Chengdu 610041 China

Cholesteryl ester transfer protein (CETP) is a key participant in the reverse transport of cholesterol from the peripheral tissues to the liver. To understand the role that CETP gene plays in the pathogenesis of coronary heart disease (CHD) , the promoter region, all 16 exons and adjacent intronic regions of CETP gene were screened for single nucleotide polymorphisms (SNPs) in 203 CHD patients and 209 controls by a combination of PCR, denaturing high performance liquid chromatography (DHPLC), molecular cloning, and DNA sequencing. A novel missense mutation in the CETP gene was identified. This mutation (L296Q) was a T-to-A conversion at codon 296 of exon 10 which replaced the codon for leucine (CTG) with the codon for glutamine (CAG). Association study revealed that L296Q mutation was associated with CHD with a significantly higher mutant allele frequency in the CHD patients than that in the controls (0. 160 vs. 0.091,x2= 9.014, P = 0.003), and that the odds ratio for the development of CHD was 1.83 for

关键词： cholesteryl ester transfer protein gene (CETP gene) coronary heart disease (CHD) missense mutation plasma lipids single nucleoti.de polymorphism (SNP)

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Effects of lanthanum and gadolinium on proliferation and differentiation of primary osteoblasts

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Progress in Natural Science:Materials International 2007年第5期17卷 618-623页

作者： Zhang Dawei , Zhang Jinchao , Chen Yao , Yang Mengsu and Yao Xinsheng (department of Natural Products Chemistry, Shenyang Pharmaceutical university, Shenyang 110016, china shenzhen Research Institute of City university of Hong Kong, shenzhen 518057, china department of Biology and Chemistry, City university of Hong Kong, china department of Chemistry, College of Chemistry & Environmental Science, Hebei university, Baoding 071002, china Key laboratory for Research and Development of Traditional Chinese medicine and Natural medicine at shenzhen, shenzhen 518057, china) department of Natural Products Chemistry Shenyang Pharmaceutical University Shenyang 110016 China shenzhen Research Institute City University of Hong Kong Shenzhen 518057 Hong Kong department of Biology and Chemistry City University of Hong Kong Hong Kong Hong Kong department of Chemistry College of Chemistry and Environmental Science Hebei University Baoding 071002 China Key Lab. for R and D of Traditional Chinese medicine and Natural medicine at shenzhen Shenzhen 518057 China

The effects of La3+ and Gd3+ on the proliferation, differentiation and adipogenic transdifferentiation of rat calvarial osteoblast-like cells (ROB cells) were evaluated by MTT method, measuring the activity of alkaline phosphatase (ALP) and Oil red O measurement. Both of La3+ and Gd3+ inhibited the proliferation of ROB cells at all concentrations (1×10-5, 1×10-6, 1×10-7, 1×10-8, 1×10-9 mol·L-1). La3+ at concentration of 1×10-5mol·L-1 significantly increased the alkaline phosphatase activity of ROB cells up to 3 folds (P<0.01). However, the effects reversed to inhibit at other concentrations. Gd played a negative role in the alkaline phosphatase activity. La3+ inhibited the adipogenic transdifferentiation of ROB cells at all concentrations in a dose-dependent way. However, Gd promoted the adipogenic transdifferentiation of ROB cells at 1×10 and 1×10 mol·L . These findings suggested that the effects of rare earth elements on the proliferation, differentiation and adipogenic transdifferentiation of ROB cells were dependent on their concentrations and species.

关键词： lanthanum gadolinium osteoblast-like cells proliferation differentiation transdifferentiation.

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Lactate Dehydrogenase Isoenzyme of Lens in Fetus

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眼科学报 2002年第3期18卷 194-196页

作者： Kexi Ding, Yasong Zhangdepartment of Ophthalmology, peking university.shenzhen hospital, shenzhen 518036, chinadepartment of laboratory, Guangzhou Liuhua Bridge hospital, Guangzhou 510000, china department of Ophthalmology Peking University Shenzhen Hospital Shenzhen 518036 China

Purpose: The LDH isoenzyme patterns were evaluated in lens of fetus to study the metabolism in human ***: LDH isoenzyme electrophoresis was performed in 15 lenses of human fetus aged from 4～6 ***: There are 5 bands of LDH in fetus lens. LDH4 lever is the highest, and LDH1 is the lowest, especially in the cortex of lens. LDH1 is higher in lens of 4 months of age than in that of 5 and 6 months of ***: LDH isoenzyme in lens of fetus shows mainly anaerobic metabolism in development, but there is also aerobic metabolism, especially in younger fetus.

关键词：胎儿晶状体乳酸脱氢酶同工酶

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Transforming growth factor-β1 gene polymorphisms associated with chronic obstructive pulmonary disease in Chinese population

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Acta Pharmacologica Sinica 2005年第6期26卷 714-720页

作者： Zhi-guang SU~2, Fu-qiang WEN, Yu-lin FENG, Min XIAO, Xiao-ling WU laboratory of Pulmonary medicine, west china hospital, sichuan university Key laboratory of Biotherapy of Human Disease, Ministry of Education, chengdu 610041, china laboratory of Pulmonary medicine West China Hospital Sichuan University Key laboratory of Biotherapy of Human Disease Ministry of Education Chengdu China

Aim: To determine the frequencies of polymorphism and haplotype in the transforming growth factor-beta 1 （TGF-1） gene promoter in the Chinese population and to investigate the susceptibility of this population to chronic obstructive pulmonary disease （COPD）. Methods: The target fragments of the TGF-1 gene promoter were amplified and analyzed by polymerase chain reaction-restriction fragment length polymorphism technique in 84 COPD patients and 97 age- and sex-matched healthy controls. The test for Hardy-Weinberg equilibrium was performed using HWE program of the LINKUTIL package and statistical analysis was carried out with the SPSS statistical package. An expectation maximization algorithm was used for the pairwise linkage disequilibrium test and haplotype analysis. Results: More carriers of the -800A allele, or fewer carriers of the -509T allele, were detected in the COPD patients compared with the non-symptomatic control subjects [for the -800A allele, 29.8% vs 14.4%, respectively, x2=6.257, degrees of freedom （df）=l, P=0.012; for the -509T allele, 27.3% vs 44.3%, respectively,x2=5.582, df=l, P=0.018]. The prevalence of the -800A allele was significantly higher in the COPD patients than in control subjects OP=0.009), whereas the frequency of the -509T allele was significantly higher in control subjects than in the COPD patients （P=0.008）. In addition, this distribution tendency for the -800A or -509T allele was similar in heavy smokers （smoking history 鈮?0 pack years）; （number of packs of cigarettes per day multiplied by the number of years of smoking） X2=7.235, P=0.007, and X2=5.636, P=0.018, respectively). The linkage disequilibrium was found between -800 G鈫扐 and -509 C鈫扵（D>0.60,P509, was significantly higher in patients with COPD than in controls （0.056 vs 0.021, P5）. Conclusions: The single nucleotide polymorphism （SNP） in the TGF-1 g

关键词： chronic obstructive pulmonary disease transforming growth factor-β1 single nucleotide polymorphisms haplotypes

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Different Effects of Homocysteine and Oxidized Low Density Lipoprotein on Methylation Status in the Promoter Region of the Estrogen Receptor α Gene

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Acta Biochimica et Biophysica Sinica 2007年第1期39卷 19-26页

作者： Yushan HUANG~1 Kejun PENG~2 Juan SU~1 Yuping HUANG~3 Yizhon XU~1 and Shuren WANG~1 ~1department of Pathophysiology,west china School of Preclinic Medical Sciences & Forensic medicine,sichuan university,chengdu 610041.china ~2department of laboratory medicine,chengdu Medical College,chengdu 610500,china ~3department of Biochemistry and Molecular Bilogy,Gannan Medical College,Ganzhou 341000,china department of Pathophysiology West China School of Preclinic Medical Sciences & Forensic Medicine Sichuan UniversityChengdu 610041 China department of laboratory medicine Chengdu Medical CollegeChengdu 610500 China department of Biochemistry and Molecular Biology Gannan Medical CollegeGanzhou 341000 China

We investigated the effects of homocysteine (Hcy) and oxidized low density lipoprotein (ox-LDL) on DNA methylation in the promoter region of the estrogen receptor α (ERos) gene,and its potentialmechanism in the pathogenesis of *** smooth muscle cells (SMCs) of humans weretreated by Hcy and ox-LDL with different concentrations for different periods of *** DNA methylationstatus was assayed by nested methylation-specific polymerase chain reaction,the lipids that accumulated inthe SMCs and foam cell formations were examined with Oil red O *** proliferation of SMCs wasassayed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide *** results showedthat ox-LDL in moderate concentrations (10-40 mg/L) induced de novo methylation in the promoter regionof the ERα gene of ***,high concentrations (50 mg/L) of ox-LDL,resulted in demethylation ofERα.The Hcy treatment resulted in de novo methylation in the promoter region of the ERα gene with aconcentration- and treating time-dependent manner,and a dose-dependent promoting effect on *** data indicated that the two risk factors for atherosclerosis had the function of inducingde novo methylation in the promoter region of the ERα gene of SMCs. However,high concentrations (50rag/L) of ox-LDL induced demethylation,indicating that different risk factors of atherosclerosis with differentpotency might cause different aberrant methylation patterns in the promoter region of the ERα *** mechanism of Hcy might involve the hypermethylation of the ERα gene,leading to the proliferationof SMCs in atherosclerotic lesions.

关键词： atherosclerosis homocysteine oxidized low density lipoprotein estrogen receptor α DNA methylation

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Inhibiting the expression of hepatocyte nuclear factor 4 alpha attenuates lipopolysaccharide/ D-galactosamine-induced fulminant hepatic failure in mice

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Hepatobiliary & Pancreatic Diseases International 2012年第6期11卷 624-629页

作者： En-Qiang Chen, Dao-Yin Gong, Xiao-Hua Leng, Lang Bai, Cong Liu, Li-Chun Wang , Hong Tang Center for Infectious Diseases, west china hospital and State Key laboratory of Biotherapy ,department of Forensic pathology, College of Basic medicine and Forensic medicine , sichuan university, chengdu 610041, china Center for Infectious Diseases West China Hospital and State Key Laboratory of Biotherapy Sichuan University Chengdu 610041 China department of Forensic pathology College of Basic Medicine and Forensic Medicine Sichuan University Chengdu 610041 China

BACKGROUND: Hepatocyte nuclear factor 4 alpha (HNF4α) plays an important role in regulating cytokine-induced inflammatory responses. This study aimed to investigate the role of HNF4α in the development of fulminant hepatic failure (FHF) induced by lipopolysaccharide/D-galactosamine (LPS/D-GalN). METHODS: The FHF model was induced by simultaneous intraperitoneal injection of LPS/D-GalN in mice. Three days prior to LPS/D-GalN administration, HNF4α short-hairpin interfering RNA expression plasmid or physiological saline was injected via the tail vein with a hydrodynamics-based procedure. The degree of hepatic damage and cumulative survival rate were subsequently assessed. RESULTS: The expression of HNF4α was increased in the early stage after LPS/D-GalN administration. Inhibiting the expression of HNF4α reduced serum levels of alanine aminotransferase and aspartate aminotransferase, alleviated histological injury, and improved the survival of mice with FHF. In addition, both serum and hepatic tumor necrosis factor alpha expression were suppressed when HNF4α expression was inhibited in mice with FHF. CONCLUSION: Inhibiting HNF4α expression protects mice from FHF induced by LPS/D-GalN, but the exact mechanism behind this needs further investigation.

关键词： hepatocyte nuclear factor 4α short-hairpin RNA fulminant hepatic failure lipopolysaccharide D-galactosamine

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Decreased expression of human aquaporin-5 correlated with mucus overproduction in airways of chronic obstructive pulmonary disease

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Acta Pharmacologica Sinica 2007年第8期28卷 1166-1174页

作者： Ke WANG Yu-lin FENG Fu-qiang WEN~2 Xue-rong CHEN Xue-mei OU Dan XU Jie YANG Zhi-ping DENG Division of Pulmonary Diseases,State Key laboratory of Biotherapy of china,and department of Respiratory medicine,west china hospital of sichuan university,sichuan 610041,china Division of Pulmonary Diseases and Department of Respiratory Medicine State Key Laboratory of Biotherapy of China West China Hospital of Sichuan University Sichuan China

Aim:To investigate the relationship between aquaporin-5(AQP5)expression andmucus overproduction in the airways of Chinese patients with chronic obstruc-tive pulmonary disease(COPD)and the correlation with pulmonary ***:Bronchial tissues were obtained from fiberoptic bronchoscopyand bronchial biopsy in west china hospital from April to July ***-fivepatients were diagnosed as COPD patients,and another 20 were diagnosed as thecontrol *** expressions ofAQP5,mucin 5AC(MUC5AC),and mucin inbronchial tissues were detected by semiquantitative RT-PCR,in situ hybridization,and immunohistochemical and alcian blue-periodic acid-Schiff(AB-PAS)staining,***:Compared with the control group,an attenuated expres-sion of AQP5was detected throughout the bronchial tissues from patients withCOPD(P0.05).Simultaneously,increased staining of MUC5AC and mucus in submucosal glandswere noted(P5and increased the staining of MUC5AC and mucus in submucosal glands in theCOPD groups(P0.05).The decreased expression of AQP5mRNA was correlatedwith forced expiratory volume in 1 s(FEV1)/forced vital capacity(FVC)(r=0.60,P60,P50%vital capacity(V50)% predicted value(r=0.55,P5% vital capacity(V25)% predicted value(r=0.45,P5AC mRNA of the epitheliumairways(r=-0.45,P61,P5AC mRNA correlated with the posi-tively AB-PAS-stained area of submucosal glands and correlated negativelywith FEV1/FVC(r=-0.53;P53;P50%predicted value(r=-0.48;P5% predicted value(r=-0.4

关键词： aquaporin-5 mucin 5AC mucin mucus overproduction chronic obstructive pulmonary disease

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Analysis of CD8^+CD28^-T-suppressor cells in living donor liver transplant recipients

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Hepatobiliary & Pancreatic Diseases International 2009年第3期8卷 241-246页

作者： Yi-Xin Lin,Lan-Lan Wang,Lu-Nan Yan,Pei Cai,Bo Li,Tian-Fu Wen and Yong Zeng Center of Liver Transplantation and department of Surgery,Division of Clinical Immunology and department of laboratory medicine,west china hospital.sichuan university,chengdu610041,china Center of Liver Transplantation and department of Surgery West China Hospital Sichuan University Chengdu China.

BACKGROUND:Human CD8 + CD28 - T-suppressor(Ts) cells have been considered to indicate a reduced need for immunosuppression in pediatric liver-intestine transplant recipients and recipients of deceased heart-kidney ***,in adult-to-adult living donor liver transplantation(A-A LDLT)little information is available and the clinical significance is still unknown. METHODS:Flow cytometry was used to detect the population of CD8+CD28 -Ts cells present in peripheral blood in A-A LDLT recipients(n=31),patients with end- stage liver disease(n=24)and healthy controls(n=19). Meanwhile,we tested the graft function and trough levels of immunosuppression in *** clinical and follow- up data of 31 transplant recipients were analyzed. RESULTS:Compared with diseased controls(P=0.007) and healthy individuals(P=0.000),a notable expansion of CD8 + CD28 - Ts cells was found in recipients of A-A *** was associated with graft function,levels of immunosuppression and rejection episodes. CONCLUSIONS:To monitor the CD8 + CD28 - Ts cells levels is important to evaluate the immune state of recipients. Meanwhile,it is also important to promote expansion of CD8+CD28 -Ts cells in recipients of A-A LDLT,not only to sustain good graft function and decrease the dosage of immunosuppressants,but also to reduce the occurrence of rejection.

关键词： T-suppressor cells CD8-positive living donor liver transplantation clinical analysis

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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 730000,china ~3 department of Gynaecology andObstetrics,peking university Third hospital,Beijing 100083,china ~4 Institute of Vascular medicine,peking university Third hospital andKey laboratory of Molecular Cardiovascular Sciences Ministry of Education,Beijing 100083,china 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 *** 33258 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 24,48,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 24,48,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 24,36,and 48 ***,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

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