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Differentiation of embryonic stem cells into insulin-producing cells promoted by Nkx2.2 gene transfer

World Journal of Gastroenterology 2005年第27期11卷 4161-4166页

作者： Akira Shiroi Shigehiko Ueda Yukiteru Ouji Ko Saito Kei Moriya Yuko Sugie Hiroshi Fukui Shigeaki Ishizaka Masahide Yoshikawa Division of Developmental Biology Department of Gastroenterology and Hepatology

AIM： To investigate the ability of a genetically altered embryonic stem （ES） cell line to generate insulin-producing cells in vitro following transfer of the Nkx2.2 ***： Hamster Nkx2.2 genes were transferred into mouse ES cells. Parental and Nkx2.2-transfected ES cells were initiated toward differentiation in embryoid body （EB） culture for 5 d and the resulting EBs were transferred to an attached culture system. Dithizone （DTZ）, a zincchelating agent known to selectively stain pancreatic beta cells, was used to detect insulin-producing *** outgrowths were incubated in DTZ solution （final concentration, 100μg/mL） for 15 rain before being examined microscopically. Gene expression of the endocrine pancreatic markers was also analyzed by RT-PCR. In addition, insulin production was determined immunohistochemically and its secretion was examined using an ***： DTZ-stained cellular clusters appeared after approximately 14 d in the culture of Nkx2.2-transfected ES cells （Nkx-ES cells）, which was as much as 2 wk earlier, than those in the culture of parental ES cells （wt-ES）. The frequency of DTZ-positive cells among total cultured cells on day 28 accounted for approximately 1.0% and 0.1% of the Nkx-ES- and wt-ES-derived EB outgrowths, respectively. The DTZ-positive cellular clusters were found to be immunoreactive to insulin, while the gene expressions of pancreatic-duodenal homeobox 1 （PDX1）, proinsulin 1 and proinsulin 2 were observed in the cultures that contained DTZ-positive cellular *** secretion was also confirmed by ELISA, whereas glucose-dependent secretion was not ***： Nkx2.2-transfected ES cells showed an ability to differentiate into insulin-producing cells.

关键词： Animals Cell Differentiation Cell Line Cricetinae Gene Transfer Techniques Homeodomain Proteins Insulin Islets of Langerhans mice mice, inbred strains Stem Cells Transcription Factors

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Fractionated radioimmunotherapy using low doses of iodine 131 labeled anti-CEA monoclonal antibody after tumor volume reduction

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Chinese Medical Journal 2000年第1期113卷 53-58页

作者：田嘉禾丁勇张锦明曹丽敏尹大一 Department of Nuclear Medicine Great Wall Hospital Beijing 100853 China Department of Nuclear Medicine Chinese PLA General Hospital Beijing 100853 China

OBJECTIVE: To assess the efficacy of fractionated administration of radiolabeled monoclonal antibody in the treatment of metastases after tumor volume reduction surgery, various experimental therapies were studied comparatively. METHODS: A total of 200 inbred mice received tumor implantation from a murine adenocarcinoma cell line. The mice were randomly grouped to give saline, Arc-a, 131I-C50 in single or fractionated doses, cold C50, or non-specific 131I-IgG with or without surgical removal of the implanted tumor xenograft. RESULTS: In comparison to controls, animals receiving Arc-a and radioactive agents had longer survival, smaller tumor, better clinical condition, and less metastases foci. The best therapeutic response was noted after fractionated doses of 131I-C50, which showed better results in every aspect than those treated with other modalities. The favorable outcome was even more pronounced after tumor volume reduction. CONCLUSIONS: Fractionated dosing may improve the deposition of radiolabeled monoclonal antibody (McAb) and provide the best therapeutic effect on implanted tumor and metastases. Thus fractionated radioimmunotherapy (RIT) after tumor volume reduction might be a practical method with promising therapeutic results.

关键词： Radioimmunotherapy Animals Antibodies, Monoclonal Body Weight Carcinoembryonic Antigen Dose Fractionation Iodine Radioisotopes mice mice, inbred strains Neoplasm Metastasis Neoplasms, Experimental Radiotherapy Dosage

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Molecular mechanism about lymphogenous metastasis of hepatocarcinoma cells in mice

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World Journal of Gastroenterology 2001年第4期7卷 532-536页

作者： Li Hou Ying Li Yong-Hua Jia Bo Wang Yi Xin Mao-Ying Ling Shen Lü Department of Pathology,Dalian Medical University,Dalian 116027,Liaoning Province,ChinaDepartment of Biochemistry,Dalian Medical University,Dalian 116027,Liaoning Province,China Department of Pathology Dalian Medical University Dalian 116027 China. houli72@***

AIM: To investigate the correlation between lymphogenous metastasis and matrix metalloproteinases (MMPs) activity and the expression of Fas ligand of tumor cells in lymph nodes. METHODS: Fifty-six inbred 615-mice were equally divided into 2 groups and inoculated with Hca-F and Hca-P cells. Their lymph node metastatic rates were examined. Growth fraction of lymphocytes in host lymph nodes was detected by flow cytometry. The Hca-F and Hca-P cells were cultured with extract of lymph node, liver or spleen. The quantity of MMPs in these supernatants was examined by zymographic analysis. The expression of Fas ligand, PCNA, Bcl-2 protein of Hca-F and Hca-P cells in the mice were examined by immunohistochemistry. The apoptosis signals of macro-phages in lymph nodes were observed with in situ DNA fragmentation. RESULTS: On the 28th day post-inoculation, the lymph node metastatic rate of HcaF was 80%(16/20), whereas that of Hca-P was 25%(5/20). The growth fraction of lymphocytes was as follows: in the Hca-F cells, the proliferating peak of lymphocytes appeared on the 14th day post inoculation and then decreased rapidly, while in HcaP cells, the peak appeared on the 7th day post inoculation and then kept at a high level. With the extract of lymph node, the quantity of the MMP-9 activity increased (P<0.01) and active MMP-9 and MMP-2 were produced by both Hca-F and Hca-P tumor cells, which did not produce MMPs without the extract of lymph node or with the extracts of the liver and spleen. The expression of Fas Ligand of Hca-F cells was stronger than that of Hca-P cells (P <0.01). The expressions of PCNA and Bcl-2 protein of Hca-F cells in the tumors of inoculated area were the same as that of Hca-P cells. In situ DNA fragmentation showed that the positive signals of macrophages were around Hca-F cells. CONCLUSION: Secretion of MMPs which was associated with metastatic ability of Hca-F and Hca-P tumor cells depends on the environment of lymph nodes. The increased expression

关键词： Animals DNA Fragmentation Flow Cytometry Gelatinase A Gelatinase B Liver Neoplasms, Experimental Lymph Nodes Lymphatic Metastasis Macrophages Membrane Glycoproteins mice mice, inbred strains Research Support, Non-U.S. Gov't Tumor Cells, Cultured

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Protective effects of cyclosporine A on T-cell dependent ConA-induced liver injury in Kunming mice

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World Journal of Gastroenterology 2001年第4期7卷 569-571页

作者： Xiu-Li Zhang Qi-Zhen Quan Zi-Qin Sun Yao-Jun Wang Xue-Liang Jiang Dong-Wang Wen-Bo Li Department of Gastroenterology,General Hospital of Jinan Military Command,Jinan 250031,Shandong Province,China Department of Gastroenterology Chinese PLA General Hospital of Jinan Command Area Jinan 250031 Shandong Province China. xiuliz@***

INTRODUCTIONThe T-cell dependent specific liver injury in mice induced by concanavalin A(ConA) is a newly cstablished experimental liver injury model,which is considered more eligible for the study on pathophysiology of several human liver discascs,such as viral hepatitis and autommune hepatitis[1-9].T cell activation and several cytokines release had been proven to play a critical role in ConA -induced liver injury[10-19].Cyclosprine A(CsA),an effective inhibitor of activation of T lymphocytc,hes been used widely in clinical treatment,especially in autoimmune diseases and organ transplantation[20-25].In this study,we investigated the possible effect of CsA on ConA-induced liver injury in Kunning mice.

关键词： Animals Concanavalin A Cyclosporine Disease Models, Animal Immunosuppressive Agents Liver Liver Diseases Male mice mice, inbred strains T-Lymphocytes Tumor Necrosis Factor-alpha

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Experimental study on antitumor effect of arsenic trioxide in combination with cisplatin or doxorubicin on hepatocellular carcinoma

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World Journal of Gastroenterology 2001年第5期7卷 702-705页

作者： Wei Wang~1 Shu-Kui Qin~1 Bao-An Chen~2 Hui-Ying Chen~1 1 Chinese PLA Cancer Center,Chinese PLA 81 Hospital,Nanjing 210002,Jiangshu Province,China2 Affliliated Zhongda Hospital of Southeast University Medical College,Nanjing 210087,Jiangsu Province,China Chinese PLA Cancer Center Chinese PLA 81 Hospital Nanjing 210002 Jiangshu Province China.

INTRODUCTIONThe main component of a traditional Chinese drug 'Pishuang'. arsenic trioxide (As2O3), has obviously selective anti-tumor effect on human hepatocellular carcinoma (HCC)in both in vitro and in vivo studies[1-5]. Due to limited effectiveness when any anti-carcinogen is used alone and obviously increased toxicity when the dose is raised, there is no exception for As2O3. Furthermore, combined chemotherapy contributes to improve therapeutic effectiveness, disperse toxicity and surmount drug-resistance,in which the combination of traditional Chinese and modern medicine has more advantages and characteristics. As a result,we made an experimental study on anti-tumor effect of As2O3in combination with cisplantin (PDD) or doxorubicin (ADM)on HCC. to investigate the possibility of AS2O3 in combination with PDD or ADM and nature of interaction between them,and to provide experimental basis for clinical application.

关键词： Animals Antineoplastic Agents Antineoplastic Combined Chemotherapy Protocols Arsenicals Carcinoma, Hepatocellular Cisplatin Doxorubicin Female Humans Liver Neoplasms, Experimental Male mice mice, inbred strains Neoplasm Transplantation Oxides Research Support, Non-U.S. Gov't Tumor Cells, Cultured

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