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Telomere and telomerase in the initial stage of immortalization of esophageal epithelial cell

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World Journal of Gastroenterology 2002年第2期8卷 357-362页

作者： Zhong-Ying Shen Li-Yan Xu Wei-Jia Cai Min-Hua Chen Jian Shen,Department of Tumor pathology,medical college of shantou university,shantou 515031,Guangdong province,china En-Min Li,Department of Biochemistry and Molecular Biology,medical college of shantou university,shantou 515031,Guangdong province,china Yi Zeng,Institute of Virology,Chinese Academy of preventive Medicine,Beijing 100052,china Department of Tumor pathology Medical College of Shantou University 22 Xinling Road Shantou 515031 Guandong Province China. zhongyingshen@***

AIM: To search for the biomarker of cellular immortalization, the telomere length, telomerase activity and its subunits in cultured epithelial cells of human fetal esophagus in the process of immortalization. METHODS: The transgenic cell line of human fetal esophageal epithelium (SHEE) was established with E(6)E(7) genes of human papillomavirus (HpV) type 18 in our laboratory. Morphological phenotype of cultured SHEE cells from the 6th to 30th passages, was examined by phase contrast microscopy, the telomere length was assayed by Southern blot method, and the activity of telomerase was analyzed by telomeric repeat amplification protocol (TrAp). Expressions of subunits of telomerase, hTr and hTErT, were assessed by rT-pCr. DNA content in cell cycle was detected by flow cytometry. The cell apoptosis was examined by electron microscopy (EM) and TUNEL label. rESULTS: SHEE cells from the 6th to 10th passages showed cellular proliferation with a good differentiation. From the 12th to the 16th passages, many senescent and apoptotic cells appeared, and the telomere length sharply shortened from 23kb to 17kb without expression of hTErT and telomerase activity. At the 20th passage, SHEE cells overcame the senescence and apoptosis and restored their proliferative activity with expression of telomerase and hTErT at low levels, but the telomere length shortened continuously to the lowest of 3kb. After the 30th passage cells proliferation was restored by increment of cells at S and G2M phase in the cell cycle and telomerase activity expressed at high levels and with maintenance of telomere length. CONCLUSION: At the early stage of SHEE cells, telomeres are shortened without expression of telomerase and hTErT causing cellular senescence and cell death. From the 20th to the 30th passages, the activation of telomerase and maintenance of telomere length show a progressive process for immortalization of esophageal epithelial cells. The expression of telomerase may constitute a biomark

关键词： Cell Transformation, Neoplastic Apoptosis Cell Division Cell Line Cell Size Epithelial Cells Esophagus Humans In Situ Nick-End Labeling papillomavirus, Human research Support, Non-U.S. Gov't Telomerase Telomere

Non-wandering points and the depth for graph maps

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Science china Mathematics 2007年第12期50卷 1818-1824页

作者： Jie-hua MAI~1 Tai-xiang SUN~(2+) ~1 Institute of Mathematics,shantou university,shantou 515063,china ~2 college of Mathematics and Information Sciences,Guangxi university,Nanning 530004,china Institute of Mathematics Shantou University Shantou China college of Mathematics and Information Sciences Guangxi University Nanning China

Let G be a graph and f:G→G be *** by r(f) andΩ(f) the set of recurrent points and the set of non-wandering points of f ***Ω_0(f) = G andΩ_n(f)=Ω(f|_(Ω_(n-1)(f))) for all n∈*** minimal m∈NU {∞} such thatΩ_m(f)=Ω_(m+1)(f) is called the depth of *** this paper,we show thatΩ_2 (f)=(?) and the depth of f is at most ***,we obtain some properties of non-wandering points of f.

关键词： graph map, non-wandering point, the depth of a graph map

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Inhibition of COX-1-mediated endothelium-dependent contraction by celecoxib in mouse arteries

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中国病理生理杂志 2012年第11期 2054-2055页

作者： LIU Bin,LUO Wen-hong,LI Hui,ZHANG Ying-zhan,ZHU Ning-xia,ZHOU Ying-bi(shantou university medical college,shantou 515041,china)

AIM:Some cyclooxygenase-2(COX-2)inhibitors have been suggested to cause increased incidence of cardiovascular ***,celecoxib,a COX-2 inhibitor that is still in clinical usage,has been showed to be less detrimental to t... 详细信息

关键词： COX Inhibition of COX-1-mediated endothelium-dependent contraction by celecoxib in mouse arteries

Induction of mucin secretion from human bronchial tissue and epithelial cells by rhinovirus and lipopolysaccharide

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Acta pharmacologica Sinica 2004年第9期 74-79页

作者： Shao-heng HE, Jian ZHENG, Ming-ke DUANAllergy and Inflammation research Institute, shantou university medical college, shantou 515031, china

AIM: To examine the effects of rhinovirus and lipopolysaccharide (LpS) on mucin secretion from bronchial tissue and epithelial cells in vitro. METHODS: Human small bronchial tissue fragments (HSBTF) and human bronchial epithelial cells (HBEC) were cultured with rhinovirus 16 and LpS, respectively and culture supernatants were collected for mucin measurement. To determine mucin levels in the culture supernatants, a MUC5AC enzyme linked immunosorbent assay and an enzyme linked lectin assay procedure with dolichos biflorus agglutinin (DBA) were developed, and mucin release was expressed as percentage increased (or decreased) secretion over baseline level. rESULTS: A concentration-dependent release of DBA mucin and MUC5AC mucin were observed when HSBTF were infected with various concentrations of rhinovirus 16 at 37℃. The maximum-induced DBA mucin and MUC5AC mucin release were approximately 258 % and 83 % over baseline. The response of HSBTF to rhinovirus was completely abolished by metabolic inhibitors. rh

关键词： rhinovirus lipopolysaccharides dolichos biflorus agglutinin mucins MUC5AC mucin bronchi epithelium human

Urotensin Ⅱ accelerates cardiac fibrosis and hypertrophy of rats induced by isoproterenol

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Acta pharmacologica Sinica 2007年第1期28卷 36-43页

作者： Yong-gang ZHANG~(2,5) Yu-guang LI~2 Bao-guo LIU~3 rui-hong WEI~2 Dong-ming WANG~2 Xue-rui TAN~2 Ding-fang BU~4 Yong-zheng pANG~4 Chao-shu TANG~42 Department of Cardiovascular Diseases,First Affiliated Hospital,shantou university medical college,shantou 515041,china 3 The Affiliated Hospital,medical School of Hebei university of Engineering,Hebei 056029,china,4 Institute of Cardiovascular research,peking university First Hospital,Beijing 100034,china Department of Cardiovascular Diseases First Affiliated Hospital Shantou University Medical College Shantou China The Affiliated Hospital Medical School of Hebei University of Engineering Hebei China Institute of Cardiovascular research Peking University First Hospital Beijing China

Aim:To study whether urotensin II（UII）,a potent vasoconstrictive peptide,isinvolved in the development of cardiac hypertrophy and fibrogenesis of ratsinduced by isoproterenol（ISO）.Methods:Thirty male Wistar rats were randomlydivided into 3 *** 1 was the healthy control group,group 2 was the ISOgroup,and group 3 was the ISO+UII *** groups 2 and 3,ISO(5 mg-kg-1.d-1)wasgiven（sc）once daily for 7 *** 3 was also given UII in the first day [3 nmol/kg（5μg/kg）,iv],followed by sc（1.5 μg/kg）twice *** 1 received 0.9% *** receptor（UT）mrNA expression was determined by *** contents ofUII and angiotensin II（Ang II）were determined by *** vitro,the effects of UII on DNA/collagen synthesis of cardiac fibroblasts were deter-mined by [3H]thymidine/[3H]proline ***:The ratio of heartweight/body weight,plasma lactate dehydrogenase activity,myocardialmalondialdehyde and hydroxyproline concentration increased significantly in theISO group,as well as UT mrNA expression,plasma and cardiac UII and ventricu-lar Ang II,compared with the control group（prdial ***,UII+ISO co-treatment significantly increasedthe changes of biochemical markers of injury and the degree of cardiac hypertro-phy and *** vitro,5×10～9-5×10-7mol/L UII stimulated[3H]thymidine/[3H]proline incorporation into cardiac fibroblasts in a dose-dependent manner（presults suggest that UII was involved in the development ofcardiac fibrosis and hypertrophy by synergistic effects with ISO.

关键词： cardiac remodeling cardiac fibrosis Gpr14 receptor urotensin II collagen

Antitumor activities of human dendritic cells derived from peripheral and cord blood

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World Journal of Gastroenterology 2002年第1期8卷 87-90页

作者： Jin-Kun Zhang Jun Li Hai-Bin Chen Jin-Lun Sun Yao-Juan Qu Juan-Juan Lu Cancer pathology Laboratory,shantou university,medical college,shantou 515031,Guangdong province,china Cancer pathology Laboratory Shantou University Medical College 22 Xinlinglu Shantou 515031 Guangdong Province China. jkzhang@***

AIM: To observe the biological specialization of humanperipheral blood dendritic cells (DC) and cord blood derivedDC and its effects on effector cells killing humanhepatocarcinoma cell line BEL-7402 in ***: The DC biological characteristics were detectedwith immunohistochemical and MTT assay. Two antitumorexperimental groups are: peripheral blood DC and cordblood DC groups. peripheral blood DC groups used LAKcells as the effector cells and BEL-7402 as target cells, whilecord blood DC groups used CTL induced by tumor antigentwice pulsed DC as effector cells and BEL-7402 as targetcells, additional peripheral blood DC and cord blood DC areadded to observe its stimulating activities to effector *** effector's cytotoxicity to tumor cells were detected withneutral red colorimetric assay at two effector/target ratios of5:1 and 10: ***: peripheral blood DC and cord blood DC highlyexpressed HLA-ABC, HLA-Dr, HLA-DQ, CD54 and S-100protein. The stimulating activities to lymphocyteproliferation were compared between experimental groups(DC added) and control group (no DC added). In sixexperiment subgroups, the DC/lymphocyte ratio wassequentially 0.25: 100, 0.5: 100, 1: 100, 2: 100, 4: 100 and 8:100, A values(x± s) were 0.75396± 0.009, 0.84916± 0.010,0.90894± 0.012, 0.98371 ± 0.007, 1.01299 ± 0.006 and 1.20384± 0.006 in peripheral blood DC groups and 0.77650 ± 0.005,0.83008± 0.007, 0.92725 ± 0.007, 1.05990 ± 0.010, 1.15583 ±0.011, 1.22983 ± 0.011 in cord blood DC groups. A value was0.59517 ± 0.005 in control group. The stimulating activitieswere higher in experimental groups than in control group ( p＜ 0.01 ), which were increased when the DC concentrationwas enlarged ( p ＜ 0.01 ). Two differently derived DCs hadthe same phenotypes and similar stimulating activities ( p ＞0.05). In peripheral blood DC groups, the cytotoxicity (x ±s) of the LD groups (experimental groups) and L groups(control group) was 58.16% ± 2.03% (5: 1), 46.18% ±2.25% (10:l) and 38.13% ± 1.

关键词：肝癌树突状细胞抗肿瘤机制外周血脐带血

Stimulation of mucin secretion from human bronchial epithelial cells by mast cell chymase

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Acta pharmacologica Sinica 2004年第6期25卷 125-130页

作者： Shao-heng HE1,2,3, Jian ZHENG1 1Allergyand Inflammation research Institute, shantou university medicalcollege, shantou 515031, china 2ImmunopharmacologyGroup, Southampton General Hospital, Southampton, UK Allergy and Inflammation research Institute Shantou University Medical College Shantou 515031

AIM: To investigate the effect of chymase on the mucin secretion from human bronchialepithelial cells. METHODS: primarily-cultured human bronchial epithelial (pCHBE) cells and normal human bronchial epithelial (NHBE) cells were cultured with chymase or other stimulus in a mixture of bronchial epithelial growth medium (BEGM) and Dulbecco’s modified Eagle’s medium (DMEM), and the quantities of stimulatory mucin release were recorded. MUC5AC mucin was measured with an ELISA and dolichos biflorus agglutinin (DBA) mucin was determined with an enzyme linked DBA assay. rESULTS: A dose-dependent secretion of DBA mucin from pCHBE cells was observed with chymase with a maximum secretion of 98 % above baseline being achieved following 3 h incubation. The action of chymase started from 1 h, peaked at 3 h and dramatically decreased at 20 h following incubation. Chymase was able to also stimulate approximately 38 % increase in MUC5AC mucin release from pCHBE cells, and about 121 % increase in DBA mucin release from NHBE cells. A chymase inhibitor soybean trypsin inhibitor (SBTI) was able to inhibit up to 85 % chymase induced mucin release, indicating that the enzymatic activity was essential for the actions of chymase on bronchial epithelial cells. CONCLUSION: Chymase is a potent stimulus of mucin secretion from human bronchial epithelial cells. It can contribute to mucus hypersecretion process in the patients with chronic obstructive pulmonary disease or asthma.

关键词： chymase mucins mast cells epithelial cells MUC5AC protein

Experimental and clinic-opathologic study on the relationship between transcription factor Egr-1 and esophageal carcinoma

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World Journal of Gastroenterology 2001年第4期7卷 490-495页

作者： Ming-Yao Wu Mao-Huai Chen Ying-rui Liang Guo-Zhao Meng Huan-Xing Yang Chu-Xiang Zhuang Department of pathology,shantou university medical college,shantou 515031,Guangdong province,china Supported by the National Natural Science Foundation of china,No.39670298. Department of pathology Shantou University Medical College 22 Xinling Road Shantou 515031 Guangdong Province China. mywu@***

AIM: To observe the growth suppression effect of exogenous introduction of early growth response gene-1 (Egr-1 gene) on esophageal carcinoma tissue as well as on esophageal carcinoma cell line Eca109 and to explore the potential application of Egr-1 gene in gene therapy of tumor. METHODS: Eukaryotic expression vector of pCMV-Egr-1 plasmid was introduced into Eca109 cell line which expressed no Egr-1 protein originally with lipofectamine transfection method. The introduction and expression of pCMV-Egr-1 plasmid into Eca109 cell line was confirmed by G418 selection culture, pCr amplification of neogene contained in the vector, Western blot analysis and immunocytochemical analysis. The cell growth curve, soft agar colony formation rate and tumorigenicity in SCID mice were examined to demonstrate the growth suppression effect of exogenous Egr-1 gene on Eca109 cell line. The Egr-1 mrNA and Egr-1 protein were also detected in 50 surgical specimens of esophageal carcinoma by in situ hybridization and immunohistochemistry. rESULTS: Exogenous Egr-1 gene was introduced successfully into Eca109 cell line and expressed Egr-1 protein stably. The transfected Eca109 cell line grew more slowly than control Eca109 as shown by cell growth curves, the soft agar colony formation rate (4.0% vs 6.9%, p rage growth rate of tumor in SCID mice (35.5 +/- 7.6 vs 65.8 +/- 7.6, p pression level of Egr-1 mrNA and protein significantly increased in dysplastic epithelia adjacent to cancer rather than in cancer tissues (65.8% vs 20.0% by ISH and 57.9% vs 0.01). CONCLUSION: Exogenous Egr-1 gene shows the strong effect of growth inhibition in Eca109 cell line. Egr-1 in the cancer tissue shows down-regulated expression that supports the inhibited function of Egr-1 in cancer growth and suggests Egr-1 may have an important role in gene therapy of esophageal carcinoma.

关键词： Gene Expression regulation, Neoplastic Animals Blotting, Western Carcinogenicity Tests Cell Division DNA-Binding proteins Early Growth response protein 1 Esophageal Neoplasms Humans Immediate-Early proteins Mice Mice, SCID plasmids research Support, Non-U.S. Gov't Transcription Factors Transfection Tumor Cells, Cultured

relationship between ABO blood groups and carcinoma of esophagus and cardia in Chaoshan inhabitants of china

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World Journal of Gastroenterology 2001年第5期7卷 657-661页

作者： Min Su~1 Shan-Ming Lu~1 Dong-ping Tian~1 Hu Zhao~1 Xiao-Yun Li~2 De-rui Li~3 Zhi-Chao Zheng~4 1 Department of pathology,2 Tumor Hospital,3,4 Second Teaching Hospital,shantou university medical college,shantou 515031,china Department of pathology Shantou University Medical College Shantou 515031China. minsu@***

AIM:To study the relationship between ABO blood groups and carcinoma of esophagus and cardia in Chaoshan inhabitants of china, which is a unique Littoral high-risk area of esophageal carcinoma in china. The poor communication and transportation in the past has made Chaoshan a relatively closed area and kept its culture and custure of old china thousand years ago. METHODS: Data on age, sex, ABO blood type and X-ray or pathological diagnose of the patients with carcinoma of esophagus or cardia were collected from the Tumor Hospital. First Affiliated Hospital, Second Affiliated Hospital of shantou university medical college; and the Central Hospital of shantou and the Central Hospital of Jieyang. A total of 6685 patients with esophageal carcinoma (EC) and 2955 patients with cardiac cancer (CC) in Chaoshan district were retrospectively assessed for their association with ABO blood groups. rESULTS: The distribution of ABO blood groups in patients with EC or CC was similar to the normal local population in Chaoshan. However, blood group B in male patients with CC and in the patients with carcinoma in the upper third esophagus was 2.3% and 4.7% higher than the corresponding controls. The relative risk B O was 1.1415 (p2696 (prespectively. No relationship was found between ABO blood groups and tumor differentiation. CONCLUSION: ABO blood group B is associated with the incidence of CC in male individuals and carcinoma in the upper third esophagus. The distribution of ABO blood groups varies in the different geographical and ethnic groups. As a result, proper controls are very important for such studies.

关键词： ABO Blood-Group System Cardia china Esophageal Neoplasms Female Gene Frequency Genetic predisposition to Disease Humans Male research Support, Non-U.S. Gov't retrospective Studies risk Factors Sex Distribution Stomach Neoplasms