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Expression of angiostatin cDNA in human gallbladder carcinoma cell line GBC-SD and its effect on endothelial proliferation and growth

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World Journal of Gastroenterology 2006年第17期12卷 2762-2766页

作者： ding-zhong yang Jing He Ji-Cheng Zhang Zuo-Ren Wang Department of Surgery The First Hospital Xi'an Jiaotong University Xi'an 710065 Shannxi Province China Deparment of Pharmacology University of Texas Medical Branch Galveston 77555 Texas State United States Department of surgery Union Hospital Fujian Medical University Fuzhou 350001 Fujian Province China Department of surgery The First Hospital Xi' an Jiaotong University Xi'an 710065 Shannxi Province China

AIM： To explore the influence of angiostatin up-regulation on the biologic behavior of gallbladder carcinoma cells in vitro and in vitro, and the potential value of angiostatin gene therapy for gallbladder carcinoma. METHODS： A eukaryotic expression vector of pcDNA3.1（＋） containing murine angiostatin was constructed and identified by restriction endonuclease digestion and sequencing. The recombinant vector pcDNA3.1-angiostatin was transfected into human gallbladder carcinoma cell line GBC-SD with Upofectamine 2000, and paralleled with the vector and mock control. The resistant clone was screened by G418 filtration. Angiostatin transcription and protein expression were examined by RT-PCR, immunofluorescence and Western-blot. The supernatant was collected to treat endothelial cells. Cell proliferation and growth in vitro were observed under microscope. RESULTS： Murine angiostatin cDNA was successfully cloned into the eukaryotic expression vector pcDNA3.1 （＋）. AFter 14 d of transfection and selection with G418, macroscopic resistant cell cloning was formed in the experimental group transfected with pcDNA 3.1（＋）-angiostatin and vector control. But untreated cells died in the mock control. Angiostatin was detected by RT-PCR and protein expression was detected in the experimental group by immunofluorescence and Western-blot. Cell proliferation and growth in v/tro in the three groups were observed respectively under microscope. No significant difference was observed in the growth speed of GBC- SD cells between groups that were transfected with and without angiostatin. After treatment with supernatant, significant differences were observed in endothelial cell （ECV-304） growth in vitro. The cell proliferation and growth were inhibited. CONCLUSION： Angiostatin does not directly inhibit human gallbladder carcinoma cell proliferation and growth in vitro, but the secretion of angiostatin inhabits endothelial cell proliferation and growth.

关键词： Angiostatin Gallbladder carcinoma Endothelial cell

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Angiostatin inhibits pancreatic cancer cell proliferation and growth in nude mice

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World Journal of Gastroenterology 2005年第32期11卷 4992-4996页

作者： ding-zhong yang Jing He Ji-Cheng Zhang Zhuo-Ren Wang Department of Surgery The First Hospital Xi'an Jiaotong University Xi'an 710065 Shaanxi Province China Department of Pharmacology University of Texas Medical Branch Galveston 77555 Texas State USA Department of Surgery Union Hospital Fujian Medical University Fuzhou 350001 Fujian Province China Zhuo-Ren Wang Department of Surgery The First Hospital Xi'an Jiaotong University Xi'an 710065 Shaanxi Province China

AIM: To observe the biologic behavior of pancreatic cancer cells in vitro and in vivo, and to explore the potential value of angiostatin gene therapy for pancreatic ***: The recombinant vector pcDNA3.1(+)-angiostatin was transfected into human pancreatic cancer cells PC-3 with Lipofectamine 2000, and paralleled with the vector and mock control. Angiostatin transcription and protein expression were determined by immunofluorescence and Western blot. The stable cell line was selected by G418. The supernatant was collected to treat endothelial cells. Cell proliferation and growth in vitro were observed under microscope. Cell growth curves were *** troms-fected or untroms-fected cells overexpressing angiostatin vector were implanted subcutaneously into nude mice. The size of tumors was measured, and microvessel density count (MVD) in tumor tissues was assessed by immunohistochemistry with primary ***: After transfected into PC-3 with Lipofectamine 2000 and selected by G418, macroscopic resistant cell clones were formed in the experimental group transfected with pcDNA 3.1(+)-angiostatin and vector control. But untreated cells died in the mock control. Angiostatin protein expression was detected in the experimental group by immunofluorescence and Western-blot. Cell proliferation and growth in vitro in the three groups were observed respectively under microscope. After treatment with supernatant, significant differences were observed in endothelial cell (ECV-304) growth in vitro. The cell proliferation and growth were inhibited. In nude mice model, markedly inhibited tumorigenesis and slowed tumor expansion were observed in the experimental group as compared to controls, which was parallel to the decreased microvessel density in and around tumor ***: Angiostatin does not directly inhibit human pancreatic cancer cell proliferation and growth in vitro,but it inhibits endothelial cell growthin vitro. It exerts the anti-tum

关键词： Angiostatin Pancreatic cancer Endothelial cell Nude mice

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Effects of IGF-1 and oxLDL on expression of phosphatase PHLPP1 in vascular smooth muscular cells

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Journal of Geriatric Cardiology 2009年第4期6卷 237-240页

作者： Xing-Li Wu ding-You yang zhong-Su yang De-Yin Li Hui-Bin Xu Shi-Wen Wang Institute of Geriatric Cardiology Chinese PLA General Hospital Beijing 100853 China

Objective To investigate the effects of insulin-like growth factor-1 （IGF-1） and oxidized low density lipoprotein （oxLDL） on expression ofphosphatase PHLPP 1 in vascular smooth muscle cells （VSMCs）. Methods Rabbit aortic VSMCs were cultured. VSMCs proliferation ability was determined by measuring cell number and mitochondrial dehydrogenase （MD） activity with MTT assay. Western blot was used to detect the protein expression ofphosphatase PHLPP1. Results IGF-1 （100ug/L） increased cell number and MD activity to 3.02 and 3.59 times of that in control group, oxLDL（501xg/ml） elevated the above two parameters to 2.03 and 2.91 times respectively. Western blot showed that IGF-1 and oxLDL inhibited the expression of PHLPPI to 39.27% and 40.26% of the control group （P〈0.01 ）. Conclusion IGF- 1 and oxLDL may enhance the proliferation of VSMCs by decreasing the expression ofphosphatase PHLPP 1.

关键词： PH domain leucine-rich repeat protein phosphatasel (PHLPP1) insulin-like growth factor-l oxidized low density lipoprotein(oxLDL) vascular smooth muscle cells

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Types of uranium deposits in central Zhuguang Mountains in Hunan Province,South China and their metallogenic regularity and prospecting directions

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China Geology 2020年第3期3卷 411-424页

作者： Zhen-hua Xiao Shen-bang Xiong Chun-hua Li Ying Liu zhong-ding yang Xiao-xi Feng Xue-wen Liu No.302 Brigade of Hunan Nuclear Geology Chenzhou 432000China East China University of Technology Nanchang 330013China Tianjin Center China Geological SurveyMinistry of Natural ResourcesTianjin 300170China

The central Zhuguang Mountains in Hunan Province is located at the junction of the three provinces,namely Hunan,Jiangxi,and Guangdong,where the famous Lujing uranium ore field *** uranium deposits occurring in this area are all granite-related and they can be divided into three types,namely endogranitic ones,perigranitic ones,and contact zone *** endogranitic uranium deposits are mainly controlled by the structural alteration zones developing within granites,with fragmentation,hematitization,and alkali metasomatism as their main mineralization *** perigranitic uranium deposits are mainly produced in the carbonaceous,siliceous,and argillaceous composite layers of epimetamorphic rocks and are controlled by fractured zones formed due to interlayer *** contact zone type uranium deposits mainly occur in the contact parts between the granites and favorable *** have developed in favorable sections where multiple sets of structures are combined and *** main metallogenic regularities of uranium in the central Zhuguang Mountains are as *** basic conditions for the uranium mineralization in this area include the framework consisting of regional deep large faults and their associated multi-set multi-direction favorable metallogenic structures,multi-cycle and multi-stage uranium-rich rock masses,and uranium-rich folded ***,the uranium deposits in this area are closely related to granites in terms of genesis and space and they are formed in different structural parts subject to the same ***,based on the summary of the characteristics and regularities of uranium mineralization in this area,the controlling factors of different types of uranium deposits in the area were explored and six metallogenic target areas were *** these will provide references for the exploration of uranium deposits in this area.

关键词： Uranium deposits Magmatic hydrothermal type Metallogenic regularity Mineral exploration engineering Prospecting direction Zhuguang Mountain Hunan Province South China

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石蒜总生物碱的提取及其对酒精发酵过程的抑制机理

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食品科学 2011年第12期32卷 83-87页

作者：杨馨张梁丁重阳石贵阳工业生物技术教育部重点实验室江南大学生物工程学院生物资源与生物能源研究中心江苏无锡214122

基于对石蒜生物碱抑菌作用的研究,结合生物碱的药用价值,提出酒精发酵前提取生物碱的必要性。通过单因素试验探讨生物碱的提取方式和提取条件对生物碱提取率的影响,结果表明,提取剂为甲醇、料醇比1:15(g/mL)、提取温度60℃、提取时间1h... 详细信息

基于对石蒜生物碱抑菌作用的研究,结合生物碱的药用价值,提出酒精发酵前提取生物碱的必要性。通过单因素试验探讨生物碱的提取方式和提取条件对生物碱提取率的影响,结果表明,提取剂为甲醇、料醇比1:15(g/mL)、提取温度60℃、提取时间1h、提取液pH11.0条件下,生物碱的提取量达28.70mg/g;进一步对含有生物碱和不含生物碱的两种石蒜原料进行酒精发酵过程比较,结果显示,石蒜生物碱对糖化酶活性和工业酒精酵母的生长均具有明显抑制作用,对α-淀粉酶则影响较小,初步揭示了石蒜生物碱对酒精发酵过程的抑制机理。

关键词：石蒜生物碱提取酒精发酵抑制机理

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添加中药的鸡腿蘑发酵液对α-葡萄糖苷酶及非酶糖基化的抑制作用

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中国医药工业杂志 2006年第6期37卷 384-387页

作者：王锋丁重阳章克昌江南大学生物资源研究室教育部工业生物技术重点实验室江苏无锡214036

在鸡腿蘑发酵培养基中添加不同中药进行液体培养,比较对α-葡萄糖苷酶及非酶糖基化的抑制率。结果表明,添加苦瓜的鸡腿蘑发酵液的抑制率最高,分别为(41．6±1．1)％和(97．0±0．3)％。以四氧嘧啶诱发的糖尿病小鼠模型进行降血糖试验,... 详细信息

在鸡腿蘑发酵培养基中添加不同中药进行液体培养,比较对α-葡萄糖苷酶及非酶糖基化的抑制率。结果表明,添加苦瓜的鸡腿蘑发酵液的抑制率最高,分别为(41．6±1．1)％和(97．0±0．3)％。以四氧嘧啶诱发的糖尿病小鼠模型进行降血糖试验,结果表明添加苦瓜的鸡腿蘑发酵液有显著降血糖作用。

关键词：鸡腿蘑 α-葡萄糖苷酶非酶糖基化发酵

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根癌土壤杆菌产辅酶Q_(10)的补料分批发酵工艺

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过程工程学报 2011年第1期11卷 113-116页

作者：金赛徐俭张梁丁重阳石贵阳江南大学工业生物技术教育部重点实验室江南大学生物工程学院生物资源与生物能源研究中心江苏无锡214122

以根癌土壤杆菌(Agrobacterium tumefaciens Q14)为生产菌株,在15L发酵罐内对补料分批发酵生产辅酶Q10的工艺进行优化.通过逐步添加限制性营养物质,考察其对发酵的影响,获得了较优的补料分批发酵工艺,即流加葡萄糖和KH2PO4溶液,将葡萄... 详细信息

以根癌土壤杆菌(Agrobacterium tumefaciens Q14)为生产菌株,在15L发酵罐内对补料分批发酵生产辅酶Q10的工艺进行优化.通过逐步添加限制性营养物质,考察其对发酵的影响,获得了较优的补料分批发酵工艺,即流加葡萄糖和KH2PO4溶液,将葡萄糖和磷浓度分别控制在5～15g/L和50～120mmol/L,同时,在30和50h各加入5%()新鲜发酵培养基.在此工艺条件下,发酵周期为84h,菌体量达87.2g/L,辅酶Q10产量高达2166.2mg/L,生产强度为25.8mg/(Lh),比分批发酵提高了7.8倍.

关键词：根癌土壤杆菌辅酶Q10 补料分批发酵

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10种中药对灵芝液体发酵的影响

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食品与生物技术学报 2008年第2期27卷 123-126页

作者：刘媛丁重阳章克昌石贵阳食品科学与技术国家重点实验室江南大学江苏无锡214122

观察中药对灵芝菌液体发酵过程中生物量和灵芝酸的影响。结果证明金银花、淡豆豉、淡竹叶、连翘和甘草的添加对灵芝菌的生长及发酵液中灵芝酸的产生均有一定促进作用;薄荷、芦根对灵芝菌生长有促进作用,但不能促进发酵液中灵芝酸的产生。

关键词：灵芝中药发酵灵芝酸

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墨汁鬼伞液体发酵营养因子对菌丝体生长和胞外多糖产量影响的研究

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食品科学 2008年第7期29卷 291-296页

作者：毕华南丁重阳石贵阳章克昌江南大学生物工程学院工业生物技术教育部重点实验室江苏无锡214122

本研究实现了墨汁鬼伞[Coprinus atramentarius(***.)Fr.]的液体培养,通过单因素试验研究了不同碳源、氮源及无机盐对墨汁鬼伞菌丝体生长和胞外多糖产量的影响。在此基础上,通过响应曲面法(RSM)研究了各营养因子的最佳水平,得到当葡萄... 详细信息

本研究实现了墨汁鬼伞[Coprinus atramentarius(***.)Fr.]的液体培养,通过单因素试验研究了不同碳源、氮源及无机盐对墨汁鬼伞菌丝体生长和胞外多糖产量的影响。在此基础上,通过响应曲面法(RSM)研究了各营养因子的最佳水平,得到当葡萄糖、玉米粉、麸皮、KH2PO4和MgSO4·7H2O分别为13.51、17.07、7.62、2.29和2g/L时,胞外多糖最大预测值为293.73mg/L;当上述变量为10.07、18.02、11.63、1.58和4g/L时,菌丝体最大生长量为6.24g/L。

关键词：墨汁鬼伞液体发酵胞外多糖生物量响应曲面法

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富锌姬松茸胞内多糖的分离及体内抑制小鼠肝癌的研究

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食品与生物技术学报 2007年第2期26卷 43-48页

作者：张笑然丁重阳章克昌江南大学工业生物技术教育部重点实验室江苏无锡214122

研究了经DEAE-纤维素离子交换柱分离的富锌液态发酵姬松茸胞内多糖组分对小鼠肝癌实体型Heps的体内抑制效果,同时考察了柱层析过后各组分中锌元素的分布情况及给药后小鼠血清锌的变化情况。分离所得组分MP-2在5 mg/(kg.d)剂量下抑瘤率为... 详细信息

研究了经DEAE-纤维素离子交换柱分离的富锌液态发酵姬松茸胞内多糖组分对小鼠肝癌实体型Heps的体内抑制效果,同时考察了柱层析过后各组分中锌元素的分布情况及给药后小鼠血清锌的变化情况。分离所得组分MP-2在5 mg/(kg.d)剂量下抑瘤率为59.5%(p<0.001),MP-3在10 mg/(kg.d)剂量下抑瘤率为68.1%(p<0.001),两样品在10 mg/(kg.d)剂量下均能使小鼠脾指数显著提高。组分MP-2含锌率3.38‰、MP-3含锌率2.46‰,远高于组分MP-1的含锌率0.12‰;注射样品MP-2、MP-3的实验组小鼠血清锌浓度皆显著高于不给予富锌样品的阴性对照组(p<0.05)。

关键词：姬松茸多糖锌离子交换肝癌

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