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Characteristics and application of established luciferase hepatoma cell line that responds to dioxin-like chemicals

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World Journal of Gastroenterology 2003年第7期9卷 1460-1464页

作者： Zhi-Ren Zhang Shun-Qing Xu Xi Sun Yong-Jun Xu Xiao-Kun Cai Zhi-Wei Liu Xiang-Lin Tan Yi-Kai Zhou Jun-Yue Zhang Hong Yan Institute of Environmental Medicine,Tongji Medical college,Huazhong University of science and Technology,Wuhan,4.0030,Hubei Province,china Institute of Environmental Medicine Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 Hubei Province China.

AIM: To establish a luciferase reporter cell line that responds dioxin-like chemicals (DLCs) and on this basis to evaluate its characteristics and application in the determination of ***: A recombinant luciferase reporter plasmid was constructed by inserting dioxin-responsive element (DREs)and MMTV promoter segments into the pGL3-promoter plasmid immediately upstream of the luciferase gene, which was structurally demonstrated by fragment mapping analysis in gel electrophoresis and transfected into the human hepatoma cell line HepG2, both transiently and stably, to identify the inducible expression of luciferase by 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD). The time course,responsive period, sensitivity, structure-inducibility and doseeffect relationships of inducible luciferase expression to DLCs was dynamically observed in HepG2 cells stably transfected by the recombinant vector (HepG2-Luc) and compared with that assayed by ethoxyresorufin-O-deethylase (EROD) in non-transfected HepG2 cells (HepG2-wt).RESULTS: The inducible luciferase expression of HepG2-Luc cells wa s noted in a time-, dose-, and AhR-dependent manner, which peaked at 4.h and then decreased to a stable level at 14.h after TCDD treatment. The responsiveness of HepG2-Luc cells to TCDD induction was decreased with culture time and became undetectable at 10th month of HepG2-Luc cell formation. The fact that luciferase activity induced by 3, 3', 4. 4.-PCB in HepG2-Luc cells was much less than that induced by TCDD suggests a structureinducibility relationship existing among DLCs. Within the concentrations from 3.5× 10-12 to 5× 10-9 mol/L, significant correlations between TCDD doses and EROD activities were observed in both HepG2-luc and HepG2-wt cells. The correlation between TCDD doses from 1.1×10-13 to 1×10-8 mol/L and luciferase activities was also found to be significant in HepG2-luc cells (r=0.997, P＜0.001), but not in their HepG2-wt counterparts. For the comparison of the enzyme respons

关键词：肝细胞癌荧光素酶二噁英类药物应用

Experimental Researches on Carcinogenesis or Tumorigenicity of MDCK Canine Kidney Cell(CKC) Lines and Analysis of Their Chromosome Karyotypes

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Agricultural sciences in china 2002年第7期1卷 816-827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, YAN Long-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 & Development of Animal Healthcare Products , Fourth Military Medical University , Xian 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, Xian 710032 , P. R . china Key Open Laboratory of Preventive Veterinary Medicine of State Ministry of Agriculture, college of Animal Medical sciences & 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 Colloge /Institute of Basic Medical sciences at the Chinese Academy of Medical sciences , Beijing 100005 , P. R . china) Department of Immunology School of Basic Medical Sciences Peking University Health Science Center Beijing 100083 P. R.China Beijing Institute of Preventive Medicine Beijing 100071P.R.China Department of Pathology at School of Pre-clinical Education Department of Toxicology at Center for Research Development of Animal Healthcare Products Fourth Military Medical University Xian 710032 P. R . China Beijing Institute of Preventive Medicine Beijing 100071P.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 Xian 710032 P. R . China Key Open Laboratory of Preventive Veterinary Medicine of State Ministry of Agriculture College of Animal Medical Sciences State Key Laboratory of Agricultural Biotechnology College of Biological Sciences China Agricultural University Beijing 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

The chromosomal number variations & structural aberrations of the MDCK cell line, primary feline or canine kidney cell(FKC or CKC) and Hela cell line were investigated and their karyotypes of conventional chromosome bands were analyzed. The carcinogenesis or tumorigenicity testing of these cell lines in about 232 nude mice and for colony formation in soft agarose and for haemagglutination under different concentration of plant lectins of these cells were carried out. 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 tetraploid YA strain of MDCK cell during 20 - 4. passages, with hy-podiploid JB strain of MDCK cell on passage 25, with di-and hypoploid JC strain of MDCK cell during 2-15 passages or with hypoploid M strain of MDCK cell during 9 - 27 passages was 28/58, 1/5, 4.18 and 0/31, 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 karyotype is, at least it has very low tumorigenicity even when its modal chromosome number is hypoploid. The repeatedly frozen, thawed and split controls of tumorigenicity-positive cell lines(X strain of Hela, M strain of BHK-21, JA strain of Vero, YA strain of MDCK) have much

关键词： Cell line MDCK Hela Feline or canine kidney cell Karyotypes Carcinogenesis or tuniori-genicity

Synthesis of a New Kind of Acyl Thiosemicarbazide

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Synthesis of a New Kind of Acyl Thiosemicarbazide

Construction of the Classification and Grading Index system of Cultivated Land Based on the Viewpoint of Sustainable Development

中国化学会第四届有机化学学术会议

作者： LI, Jian-Ping ZHENG, Peng-Zhi (college of Chemical & Environmental science, Henan Normal University, Xinxiang 4.3007)

Acyl thiosemicarbazides cause much more attention because of their herbicidal, antibacterial, anti-inflammatory and plant-growth regulating activities. Most of them are synthesized from acyl isothiocyanates and *** this paper, we report syntheses of a series of new kind of thiosemicarbazides by reaction between acyl isothiocyanates and 4.phenyl-

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Asian Agricultural Research 2010年第9期2卷 45-48,52页

作者： CUI Qian, WU Yu-ling, LI Qing college of Urban and Environmental sciences, Huazhong Normal University, Wuhan 4.0079, china

In order to objectively and reasonably evaluate the actual and potential value of cultivated land, both social and ecological values are introduced into the classification and grading index system of cultivated land based on the viewpoint of sustainable development, after considering the natural and economic values of cultivated land. Index system construction of the sustainable utilization of cultivated land should follow the principles of economic viability, social acceptability, and ecological protection. Classification of cultivated land should take into account the soil fertility of cultivated land. Then, grading of cultivated land is carried out from the practical productivity (or potential productivity) of cultivated land. According to the existing classification index system of cultivated land, the soil, natural and environmental factors in plains, mountains and hills are mainly modified in the classification index system of cultivated land. And index systems for the cultivated land classification in plains, mountains and hills are set up. The grading index system of cultivated land is established based on the economic viability (economic value), social acceptability (social value) and protection of cultivated land (ecological value). Quantitative expression of cultivated land grading index is also carried out.

关键词： Classification and grading of cultivated land Inde

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Intranet安全保障体系的分析与设计

天中学刊 2002年第2期17卷 31-33页

作者：傅丰驻马店师专计算机科学系河南驻马店463000

分析了 Intranet的安全问题 ,包括其使用的 Internet应用技术安全方面的脆弱性和企业组建 Intranet时所特有的安全需要 ,并给出了一个组建 Intranet安全保障体系的较为完整的设计方案 .

关键词： Intranet 安全保障体系网络安全体系设计体系结构安全策略企业网

利用亲和质谱技术从中药材提取物中筛选马尔堡病毒核蛋白的小分子配体

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南开大学学报（自然科学版） 2016年第5期49卷 1-7页

作者：付旭李利新汪志华王跃飞郭宇水雯箐天津科技大学生物工程学院天津300457 天津国际生物医药联合研究院高通量分子药物筛选中心天津300457 天津中医药大学中医药研究院天津300193 南开大学生命科学学院天津300071

马尔堡病毒的RNA基因组编码7种结构蛋白,其中核蛋白在基因组的复制和核衣壳形成中发挥重要作用,成为十分有吸引力的抗病毒药物潜在靶标.实验利用亲和质谱技术从甘草提取物中筛选马尔堡核蛋白的配体.首先从甘草粗提物中发现了可能与蛋白... 详细信息

马尔堡病毒的RNA基因组编码7种结构蛋白,其中核蛋白在基因组的复制和核衣壳形成中发挥重要作用,成为十分有吸引力的抗病毒药物潜在靶标.实验利用亲和质谱技术从甘草提取物中筛选马尔堡核蛋白的配体.首先从甘草粗提物中发现了可能与蛋白结合的配体甘草查尔酮A.在此基础上,对甘草中已知组分的单体混合物和甘草查尔酮A单体分别进行亲和质谱分析,验证了粗提物的质谱筛选结果.而后进一步利用SPR实验确认甘草查尔酮A单体对核蛋白靶标的结合作用,进而通过一系列生化实验发现该单体对马尔堡核蛋白热稳定性和构象均能产生功能性影响.实验发展的亲和质谱技术为从中草药提取物中直接筛选新型抗病毒药物分子提供了简便高效的新方法.

关键词：马尔堡核蛋白甘草粗提物亲和质谱配体筛选

A Scalable Intrusion Detection system for IPv6

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Wuhan University Journal of Natural sciences 2006年第6期11卷 1723-1726页

作者： LIU Bin LI Zhitang LI Zhanchun Network and Computing Center Huazhong University of Science and Technology Wuhan 430074 Hubei China

The next generation protocol IPv6 brings the new challenges to the information security. This paper presents the design and implementation of a network based intrusion detection system that support both IPv6 protocol and IPv4.protocol. This system＇s architecture is focused on performance, simplicity, and scalability. There are four primary subsystems that make it up： the packet capture, the packet decoder, the detection engine, and the logging and alerting subsystem. This paper further describes a new approach to packet capture whose goal is to improve the performance of the capture process at high speeds. The evaluation shows that the system has a good performance to detect IPv6 attacks and IPv4.attacks, and achieves 61% correct detection rate with 20% false detection rate at the speed of 100 Mb·s^-1.

关键词： intrusion detection system ipv6 packet capture

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氟化物对雏鸡细胞免疫功能的影响

畜牧与兽医 2001年第2期33卷 10-12页

作者：冯军王政富池仕红佛山科学技术学院动物医学系广东南海528231

本文研究了氟化物对雏鸡细胞免疫功能的影响。试验组Ⅰ和试验组Ⅱ在 4. 4.d分别喂含氟量为 30 0mg/kg和 50 0mg/kg的饲料 ,4.～ 56d喂常规饲料 ,对照组在整个试验期内喂常规饲料。试验结果 :在 4. 8周内 ,鸡增重减慢 ,胸腺器官指数降... 详细信息

本文研究了氟化物对雏鸡细胞免疫功能的影响。试验组Ⅰ和试验组Ⅱ在 4. 4.d分别喂含氟量为 30 0mg/kg和 50 0mg/kg的饲料 ,4.～ 56d喂常规饲料 ,对照组在整个试验期内喂常规饲料。试验结果 :在 4. 8周内 ,鸡增重减慢 ,胸腺器官指数降低 ,外周血淋巴细胞ANAE阳性率和淋巴细胞转化率显著下降。试验结果表明。

关键词：氟化物雏鸡细胞免疫抑制作用氟中毒胸腺器官指数

The relationship between loss expression of DPC4.Smad4.gene and carcinogenesis of pancreatobiliary carcinoma

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Hepatobiliary & Pancreatic Diseases International 2002年第4期1卷 624-629页

作者： Zhao-Hui Tang Sheng-Quan Zou You-Hua Hao Bao-Ju Wang Xiang-Ping Yang Qi-Qi Chen Fa-Zu Qiu the Department of General Surgery Institute of Clinical Immunology Tongji Hospital, Tongji Medical college, Huazhong University of science and Technologr, Wuhan 4.0030, china Department of Biochemistry, Rheinisch-Wistfalische Technische Hochschule (RWTH), D-52074.Aachen, Germany Department of Pathology, University of lowa, lowa city, IA5224., USA Department of General Surgery Tongji Hospital Huazhong Univ. of Science/Technology Wuhan 430030 China Institute of Clinical Immunology Tongji Hospital Huazhong Univ. of Science/Technology Wuhan 430030 China Department of Biochemistry RWTH D-52074 Aachen Germany Department of Pathology University of Iowa Iowa City IA 52242 United States

Objective: To clarify the relationship between loss of DPCA gene expression and pathogenesis of pancreato- biliary carcinoma. Methods: 75 slides of normal duct (20), hyperplasia (15), dysplasia (15), invasive carcinoma (25) from patients with pancreatic diseases including pancreatic carcinoma (25 patients), chronic pancreatitis (6), pancreas injury (2) and 71 slides of common bile duct (CBD) carcinoma (38), gallbladder carcinoma (18), hilar bile duct (HBD) carcinoma (15) from patients with primary biliary tract carcinoma were analyzed for the expression of DPC4.protein by im- munohistochemical staining. Results: All specimens from 20 cases of normal duct and 15 cases of hyperplasia showed marked expres- sion of DPC4.protein. The frequency of loss expres- sion of the DPC4.gene was 33 % in dysplasia, and 4.% in invasive carcinoma. There was a significant statistical difference between byperplasia and dyspla- sia (P4.gene was 4..3% in CBD carcinoma, 11% in gallbladder carcinoma, and 13% in HBD carcinoma. The frequency of loss expression of the DPCA gene was significantly different in CBD carcinoma vs gall- bladder carcinoma and HBD carcinoma (P4.gene occurs late in the neoplastic progression of pancreatic carci- noma. The frequency of DPC4.gene alternation was different in various locations of biliary tract carcino- ma. In CBD carcinoma, this frequency is similar to that in pancreatic carcinoma, indicating their similar molecular alternations.

关键词： DPC4.gene pancreatic carcinoma biliary tract carcinoma