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Cloning of cytochrome P-450 2C9 cDNA from human liver and its expression in CHL cells

World Journal of Gastroenterology 2002年第2期8卷 318-322页

作者： Ge-Jian Zhu Ying-Nian Yu,Department of Pathophysiology and Laboratory of Medical Molecular Biology,zhejiang university School of Medicine,hangzhou 310031,zhejiang Province,china Xin Li,Department of pharmaceutical analysis & drug metabolism,College of Pharmacology Science,zhejiang university,hangzhou 310031,zhejiang Province,china Yu-Li Qian, Present address:center of laboratory,Women’s hospital,School of Medicine,zhejiang university,hangzhou 310031,zhejiang Province,china Department of Pathophysiology and Laboratory of Medical Molecular Biology Zhejiang University School of Medicine Hangzhou 310031 Zhejiang Province China.

AIM: Using bacterial, yeast, or mammalian cell expressing a human drug metabolism enzyme would seem good way to study drug metabolism-related problems. Human cytochrome P-450 2C9(CYP2C9) is a polymorphic enzyme responsible for the metabolism of a large number of clinically important drugs. It ranks among the most important drug metabolizing enzymes in humans. In order to provide a sufficient amount of the enzyme for drug metabolic research, the CYP2C9 cDNA was cloned and expressed stably in CHL cells. METHODS: After extraction of total RNA from human liver tissue, the human CYP2C9 cDNA was amplified with reverse transcription-polymerase chain reaction (RT-PCR), and cloned into cloning vector pGEM-T. The cDNA fragment was identified by DNA sequencing and subcloned into a mammalian expression vector pREP9. A transgenic cell line was established by transfecting the recombinant vector of pREP9-CYP2C9 into CHL cells. The enzyme activity of CYP2C9 catalyzing oxidation of tolbutamide to hydroxy tolbutamide in S9 fraction of the cell was determined by high performance liquid chromatography(HPLC). RESULTS: The amino acid sequence predicted from the cDNA segment was identical to that of CYP2C9*1, the wild type CYP2C9. However, there were two base differences, i.e. 21T>C, 1146C>T, but the encoding amino acid sequence was the same, L7, P382. The S9 fraction of the established cell line metabolizes tolbutamide to hydroxy tolbutamide; tolbutamide hydroxylase activity was found to be 0.465 +/- 0.109 ***(-1).g(-1) S9 protein or 8.62 +/- ***(-1).mol(-1) CYP, but was undetectable in parental CHL cell. CONCLUSION: The cDNA of human CYP2C9 was successfully cloned and a cell line of CHL- CYP2C9, efficiently expressing the protein of CYP2C9, was established.

关键词： Cloning, Molecular Animals Aryl Hydrocarbon Hydroxylases Cell Fractionation Cell Line china Gene Expression Humans Hypoglycemic Agents Liver Protein Isoforms Recombinant Proteins Research Support, Non-U.S. Gov't Tolbutamide

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ENDOR AND ESR STUDIES OF RADICALS FORMED IN FRIEDEL-CRAFTS REACTIONS

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Science china Chemistry 1991年第11期34卷 1296-1303页

作者：桑鸿汪汉卿陈涛刘旭勤center of analysis and Research East china university of Chemical Technology Shanghai 200237 PRC Lanzhou Institute of Chemical Physics Academia Sinica Lanzhou 730000 PRC center of analysis and Research East China University of Chemical Technology

The present study deals with the study of some Friedel-Crafts alkylating system and the accurate measurement of hyperfine coupling constants by ESR and ENDOR techniques. The results indicate that the observed ESR spectra are due to polycyclic aromatic radical cations formed from their parent *** is suggested that benzyl halides produced in the Friedel-Crafts alkylating reaction undergo Scholl condensation reaction to give polycyclic aromatic hydrocarbons, which were converted into the corresponding polycyclic aromatic radical cations in the presence of aluminum chloride.

关键词： radical cations ESR ENDOR Friedel-Crafts reactions.

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A modified rat model for hepatocellular carcinoma

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Hepatobiliary & Pancreatic Diseases International 2004年第4期3卷 585-587页

作者： Fu-Chun Yang, Shu-Sen Zheng and Tian-An Jiang hangzhou, china Department of Hepatobiliary Pancreatic Surgery and Liver Transplantation center, First Affiliated Hospital, zhejiang university School of Medicine, hangzhou 310003, china Department of Hepatobiliary Pancreatic Surgery and Liver Transplantation center First Affiliated Hospital Zhejiang University School of Medicine Hangzhou 310003 China.

BACKGROUND: Rat hepatocellular carcinoma ( HCC ) model which has a high analogy to clinical liver cancer is of great value in understanding the pathogenesis and evolution of liver cancer, in searching effective anti-cancer treat- ments ( drug, hepatectomy and liver transplantation ), and designing cancer prevention strategies. In this study we es- tablished a modified rat model of hepatocellular carcinoma to enhance rats' physique and surgical endurance. METHODS: Wistar rats were fed with diethylnitrosamine (DENA) by three methods for evaluation of general condi- tions for 130 days: Doppler ultrasonographic measurement, laparotomy and histopathological examination. RESULTS: No rat died in control group ( group A) and modified DENA-induction-HCC group ( group C), but 6 deaths in classical DENA-induction-HCC group (group B) (survival rate 80%). All survived rats in groups B and C de- veloped diffusive hepatocellular carcinoma and liver cirrho- sis. General appearance of rats in the group C was better than that in the group B. CONCLUSION: With good general conditions for surgery, the modified rat model for hepatocellular carcinoma has a high carcinogenic rate and a high survival rate.

关键词： hepatocellular carcinoma model rat diethylnitrosamine

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NMR MICROSCOPY OF γ-IRRADIATED cis 1.4-POLYBUTADIENE

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Chinese Chemical Letters 1993年第7期4卷 619-620页

作者： Yi Min SHEN Xin ZHAO Lun FEI center for analysis and measurement, Fudan university, Shanghai, 200433

NMR microscopy has received great attention from various fields of research, especially material science by its possibility to study materials directly without destruction. Although this technique is almost limited to the detection of fluids constricted in solid samples, such as water in biological systems, solvent diffused in a polymer material^1 and liquid saturated rock systems^2, the mechanical defects of high elastic rubbers have also been detected^3 We are extending this technique to the direct study of the effect of y-irradiation on cis 1, 4-polybutadiene (PB). This article deals with the investigation of radiation induced crosslinking of PB by NMR microscopy.

关键词： NMR MICROSCOPY OF IRRADIATED cis 1.4-POLYBUTADIENE cis

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RHOMBUS THINKING METHOD AND ITS APPLICATION IN SCHEME DESIGN

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Chinese Journal of Mechanical Engineering 2001年第2期14卷 156-159页

作者： Zhao Yanwei Wang Wanliang Science Research center,zhejiang university of Technology Zhang Guoxian Shanghai university 浙江工业大学科学研究中心浙江杭州上海大学上海

Rhombus thinking, a new creative thinking method,is the combination of divergent thinking process and convergent thinking process,in which qualitative analysis is carried out before quantitative analysis This method tries to solve the bottle neck problem in intelligent CAD based on the extension theory The rhombus thinking method to the scheme design of new products is applied In this process, firstly, the matter element expression for the know information are set up, and then a set of matter elements are opened up by matter elements extension method; Finally,the useful information are got by appraisal method of dependent degree It has been successfully applied to the scheme design for the cutter store of machining center Theoretical and experimental results demonstrated fhat the method is much more accurate,objective and efficient than the traditional one

关键词： Extension theory Rhombus thinking method Scheme design Artificial Intelligence

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Crystal and molecular structure of guanidinium hexatungsto-bis(p-aminophenylarsonate)tetrah ydrate,[C(NH_2)_3]_4[(p-NH_3C_6H_4As)_2W_6O_(25)].4 H_2O

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Chinese Journal of Chemistry 1990年第1期8卷 37-47页

作者： LIU Ben-Yao Yih-Tong KU (GU Yi-Dong) Department of Chemistry,Fudan university,Shanghai 200433WANG Xin center of analysis and measurement,Lanzhou university,Lanzhou Department of Chemistry Fudan University Shanghai 200433 center of analysis and measurement Lanzhou University Lanzhou

The crystal structure of a new type arylarsonic polytungstate[C(NH_)_]_4[p-NH_3C_6H_4As)_2- W_6O_(25)]·4H_O was determined by single-crystal X-ray diffraction *** belongs to triclinic,space group P,with cell dimensions a=12.863(3),b=18.912(3),c=21.383(4)α=91.14(2)°,β= 93.65(3)°,γ=92.25(3)°,V=5185.9~3,Z=4,D_c=2.753g/cm^*** intensity data were collected on an Enraf-Nonius CAD4 diffractometer with Mo Kα *** positions of all tungsten and arsenic atoms were determined by direct *** other non-hydrogen atoms were revealed by difference Fourier *** structure was refined by fullmatrix least-squares procedure to a final R value of *** crystal structure contains two similar but nonidentical *** similar anions consist of a ring of six WO_6 octahedra,which are connected with one face-sharing,two corner-sharings and three edge-sharings,and two p-aminophenylarsonic tetrahedra capped above and below the ring. In each WO_6 ring,four tungsten atoms,which are joined with edge-sharing oxygen atoms,are almost coplanar,while the two others,which are joined with face-sharing oxygen atoms,protrude out of the ring towards the same *** two arsenic atoms in each anion are not equivalent in their bonding *** each anion,all non-hydrogen atoms of each organic group are in the same *** molecule contains one anion,four C(NH_2)_3^+ cations and four water *** are many hydrogen bonds between cations and anions throughout the whole *** amino groups can accept protons,so that the charge of the resulting anion decreases and[(RAs)_2W_6O_(25)]^(4-)type complexes are formed.

关键词： p-NH3C6H4As Crystal and molecular structure of guanidinium hexatungsto-bis NH2 H2O p-aminophenylarsonate)tetrah ydrate

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NUMERICAL SIMULATION METHODS AND THEIR APPLICATION IN MATERIAL HOT-WORKING

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Acta Metallurgica Sinica(English Letters) 2000年第2期13卷 494-501页

作者： J. T. Niu, H. T. Li, X. D. Meng and P. Karjaleinen 1) analysis and measurement center, Harbin Institute of Technology, Harbin 150001, china 2) Department of Mechanocal Engineering, The university of Oulu, Finland

With the development of modern science and technology, especially computer science, the numerical simulation method has been widely used in material hot-working. Mary achievements have been made in this field by using the numerical simulation method. The numerical simulation method, especially finite element method fully described in this *** of the numerical simulation method in material hot-working are also discussed. Finally, the future of the numerical simulation method is outlined.

关键词： numerical simulation material hot-working

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The construction of the full-length cDNA library of Haliotis discus hannai Ino

The construction of the full-length cDNA library of Haliotis...

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贝类学会第七次会员代表大会暨第十一次学术讨论会

作者： Xiao Liu Qi-kang Gao Guo-fan Zhang Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, Shandong, china Biomacromolecular Lab, analysis and measurement center of zhejiang university, hangzhou, 31002. zhejiang, china

Five cDNA expression library of Pacific abalone Haliotis discus hannai Ino, HD-emb, HD-met, HD-JRh, HD-RwRh and HD-tri had been established. HD-emb was constructed from the embryos sampled at gastrula and early trochophore larva. HD-met was a library focused on the metamorphosis and established from the tissue mixture of several developmental stages from trochophore larva to juvenile 11 days post-spawning. HD-JRh was from a rapid growth family at

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Synthesis and Crystal Structure of Monochloridehepta（dimethyl sulfoxide）gadolinium（Ⅲ） di（tetraphenyl boron） Salt 〔Gd（dmso）7Cl〕〔BPh4〕2

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Chinese Journal of Structural Chemistry 2000年第1期19卷 1-5页

作者： ZHANG Dao a* GUO Song Shan a WANG Han Zhang a YU Kai Bei b ( a Departmentof Chemistry, Suzhou university,Suzhou 215006 b Chengdu center of analysis and measurement, the Chinese Academy of Sciences, Chengdu 610041) 苏州大学江苏苏州 21 5006 苏州大学江苏苏州 215006 中科院成都分析测试中心四川成都 610041

The crystal of the title compound (C 62 H 82 B 2ClGdO 7S 7 M r =1378.02) is triclinic with space group P1. a=12.532(2), b=12.774(2), c=24.580(5); α=93.389(9), β=104.640(8), γ=112.923(9)°, V=3451.4(8) 3, Z=2, D c =1 326g/cm 3. μ (Mo Kα )=1.258 mm -1 , F(000)=1426. R=0.036 , R w =0.091 for 12101 reflections with I>2σ(I) . The crystal is composed of a discrete cation 〔Gd(dmso) 7Cl〕 + and two anions 〔BPh 4〕 -. The Gd(Ⅲ) ion is coordinated by a chlorine and seven oxygen atoms from seven monodenate dimethylsulfoxides formimg a distorted square antiprism coordination polyhedron.

关键词： crystal structure Gadolinium complex dimethyl sulfoxide tetraphenylboron

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Ab initio Structure Determination of [Co(NH_3)_5Br]Br_2 Using Conventional X-ray Powder Diffraction

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Chinese Chemical Letters 1997年第7期8卷 647-648页

作者： Hong Xiang WU and Li Dun MA(center for analysis and Measuremnt, Fudan university, Shanghai, 200433)(Project supported by the National Natural Science Foundation of china(NSFC) NO. 29573098) FUDAN UNIV CTR ANAL & MEASUREMENTSHANGHAI 200433PEOPLES R CHINA

The crystal structure of [Co(NH3)5Br]Br2 has been determined ab initio from the conventional X-ray powder diffraction data. The approximate structure with all 7 indeyendent non-H atoms was solved by direct methods. The final orthorhombic unit-cell parameters after Rietveld refinement are: a=13.6927, b=10.7071, c=6.9400A, V=1017.47A3, F30=93(0.0075,43), M20=49, Z=4. Space group is Pnma. The structure agreement factors are: Rp=0.066,Rwp=0.090, RF=0.041, RB=0.042.

关键词： Br2 Using Conventional X-ray Powder Diffraction Ab initio Structure Determination of NH3

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