Microbial Transformation of Gypenoside from Gynostemma pentaphyllum by Aspergullus glaucus and Its Biological Activities

以福建绞股蓝[Gynostemma pentaphyllum(Thunb)Makino]为材料,经乙醇提取和正丁醇萃取获得绞股蓝皂苷,利用灰绿曲霉(Aspergullus glaucus)微生物转化绞股蓝皂苷,通过测定绞股蓝皂苷和灰绿曲霉转化产物的抗癌、抗酪氨酸酶和抗氧化活性,比较绞股蓝皂苷经微生物转化修饰前后生物活性的差异.结果表明:灰绿曲霉转化产物对肝癌细胞SMMC7721有体外抑制作用,半抑制质量浓度(IC50)为91.66μg/m L,而绞股蓝皂苷抗癌效果不强;绞股蓝皂苷对蘑菇酪氨酸酶活力具有较强的抑制作用,IC50为0.14 mg/m L,其抑制作用属于可逆抑制,抑制类型为混合型抑制作用,而灰绿曲霉转化产物抗蘑菇酪氨酸酶活性较弱;绞股蓝皂苷和灰绿曲霉转化产物分别在0.2和2 mg/m L质量浓度下对DNA氧化损伤有保护作用.研究结果说明绞股蓝皂苷经微生物修饰后,抗癌效果增强,对蘑菇酪氨酸酶抑制作用和DNA氧化损伤保护作用减弱.该研究结果可为利用微生物转化法筛选抗癌绞股蓝皂苷奠定基础.Gynostemma pentaphyllum in Fujian Province was used as the material in this research.Gypenoside was extracted with ethanol and n-butyl alcohol. Gypenoside was microbially transformed by Aspergullus glaucus. The abilities of anticancer,anti-tyrosinase and anti-oxidation in gypenoside and transformation products were measured in our study.The difference of bioactivity between gypenoside and microbially transformed gypenoside was compared.The result showed that the products of transformation by A. glaucus affected the growth of hepatocellular carcinoma cells SMMC7721 with half maximal inhibitory concentration( IC50) values of 91. 66 μg/m L,but the anticancer activity of gypenoside was very weak.Gypenoside had strong inhibition to the mushroom tyrosinase. The value of IC50 was 0. 14 mg / m L. Moreover,the products of transformation by A. glaucus showed little anti-tyrosinase acti-vity.The kinesis study showed that the inhibition of gypenoside to mushroom tyrosinase was reversible and inhibition types were mixed. Gypenoside exhibited the active protective effect on H2O2-induced oxidative-stress damage at the concentration of 0. 2 mg / m L,and the products of transformation by A. glaucus were at 2 mg / m L. This research indicated that the anticancer effect of gypenoside modified by microorganisms was enhanced,but the activity of anti-tyrosinase and H2O2-induced oxidativestress damage became weakened.This study laid the foundation of screening anticancer gypenoside by means of microbial transformation.国家自然科学基金(81274149);; 厦门市科技创新项目(3502Z20132009,3502Z20154083);; 厦门市对台科技合作项目(3502Z20141041

溶癌灵悬乳剂肿瘤内注射的抗癌作用实验研究

目的:探讨溶癌灵合并鸦胆子种皮毒性部分研制新型肿瘤内注射药物的抗癌效果。方法:经乙醇抽提的鸦胆子种皮水溶性部分,以聚乙烯吡咯烷酮为载体,混合消癌灵制成溶癌灵悬乳剂,并以体外细胞毒杀伤及荷瘤动物肿瘤内注射抑瘤实验观察。结果:细胞毒实验显示该剂对体外培养细胞的杀伤效果与鸦胆子种皮提取液相当,但优于消癌灵及5-Fu;体内抑瘤实验表明在抑瘤作用及伤口愈合等方面,该制剂均优于消癌灵、酒精及5-Fu 组。结论:溶癌灵是一种适于肿瘤局部注射治疗的细胞毒性物质,有一定的开发前景

中药制剂局部注射的抗癌作用实验研究

近年来国内外十分重视内镜直视下治疗进展期胃癌,肿瘤内注人高浓度抗癌乳化剂,局部滞留时间长,癌组织及其所属的淋巴结内药物浓度高。我们以脂质体包载中药乳剂进行消化道肿瘤内注射,动物实验及临床应用效果明显。在此基础上,我们以鸦胆子种皮等中药提取液制成注射剂,探讨其对体外培养细胞及荷人肝癌裸鼠移植瘤模型局部注射的抗肿瘤影响。材料与方法1 材料药品:中药制剂为鸦胆子种皮、木芙蓉等中药提取液。细胞株:人肝癌细胞株 H_(9101)为厦门大学抗癌研究中心细胞培养室保存。荷人肝癌裸鼠移植瘤株 HHC_(15)由厦门大学抗癌研究中心医学实验动物室建立及保存。450型仪器微孔板

全反式维甲酸与PD98059联合应用抑制结肠癌细胞的增殖

目的本实验拟通过阐明全反式维甲酸(ATRA)与MEK1/2的特异性抑制剂PD98059联合作用对结肠癌细胞增殖的影响。方法实验分组:未加药对照组,ATRA加药组,ATRA与PD98058联合加药组。MTT法检测PD98059联合ATRA对结肠癌细胞株LS174T的增殖抑制作用。流式细胞仪检测结肠癌细胞凋亡情况。结果 MTT法显示PD98059联合ATRA应用引起细胞抑制效果明显优于单个药物作用的效果。流式细胞仪检测也表明两种药物联合应用引起结肠癌细胞凋亡效果明显优于任一单个药物。结论 ATRA与PD98059联合应用可通过引起结肠癌细胞凋亡,从而抑制癌细胞增殖

RT-PCR扩增MUC1检测胃癌微转移研究

目的　以 RT- PCR检测胃周淋巴结胃癌转移细胞并评价其临床应用价值。　方法　以 MUC1c DNA的特异序列为 RT- PCR引物 ,酶切分析法及系列稀释法对该法的特异性和敏感性进行分析 ,对临床收集的胃周淋巴结样品作 RT- PCR检测及产物 DNA点杂交验证。　结果　 ( 1)胃及胃癌组织均存在 MUC1m RNA的表达 ;( 2 )该扩增体系具有较好的特异性 ;( 3)敏感性可达 1pgRNA,相当于从 10 5个淋巴细胞中检出 1个胃癌细胞 ;( 4)对临床样品检测的结果显示较病理检查敏感 ,PCR产物点杂交进一步证实 MUC1作为 PCR标志物有较好的可靠性。　结论　该法具有较高的可靠性和较好的应用前景

纳氏测氨法半定量诊断HP感染的方法及其应用

对纳氏测氨法尿素酶试验（ＮＵＴ法）进行半定量标准化后，进一步建立了纳氏测氨半定量诊断人胃粘膜中幽门螺杆幽（ＨＰ）感染程度的方法，使ＮＵＴ法可同时用于定性及半定量诊断ＨＰ感染及感染程度．对经过半定量检测的３５０例患者作分析，显示轻、中、重度感染比率分别为５４．６％、３４．０％及１１．４％，另对其中１８２例慢性浅表性胃炎进行病变程度与ＨＰ感染程度的相关性分析，表明慢性浅表性胃炎的病变程度与ＨＰ的感染程度呈明确的正相关性（ｐ＜０．０５），提示胃部疾病的恶性演变尚与感染程度有关

成人获得性维生素K依赖性凝血因子缺乏症伴多部位出血1例

患者,男,62岁,因“反复牙龈出血,血尿3月余,腹痛,便血3 d。“为主诉入院。患者缘于入院3月余前无明显诱因始出现反复牙龈出血,偶伴血尿,无尿频、尿急、尿痛;也无发热、畏冷;曾在当地治疗,好转后出院,入院3 d前无明显诱因出现腹痛,脐周明显,伴排稀水便,初为黑色,后为鲜红色,2~3次/d,无恶心、呕吐,也无返酸、嗳气。查体:体温36.2℃、脉搏96次/MIn、呼吸20次/MIn、血

Method of Detecting Gastric Cancer Micrometastases in Lymph Nodes by Nest RT-PCR Amplification of Keration 19 and its Application

[目的]建立RT PCR扩增Keratin19mRNA的方法 ,评价其应用前景。[方法]采用Keratin19cDNA的套式引物建立巢式RT PCR扩增体系 ,以酶切分析及DNA点杂交法鉴定扩增产物的特异性 ,逆转录产物系列稀释法分析检测敏感性 ,并对51例临床胃周淋巴结样品作初步检测。[结果]该扩增体系具有较好的扩增特异性 ,检测敏感性达1pgRNA ,相当于从105 个淋巴细胞中检出1个胃癌细胞 ;对临床样品检测结果显示该法较病理检查法敏感性高。[结论]该扩增体系具有较好的特异性、敏感性和较高的可靠性。福建省卫生厅基金!(编号 :96075);福建省自然科学基金!(F00024)资

DETECTING GASTRIC CANCER MICROMETASTASES IN AXILLARY LYMPH NODES BY RT-PCR AMPLIFICATION AND SILVER STAINING

作者单位: 1 厦门大学生命科学学院抗癌中心( 厦门361005)，2 厦门大学生命科学学院生物系，3 厦门市中山医院[中文文摘]目的　建立RT PCR检测胃周淋巴结胃癌转移细胞的方法 ,评价其临床应用价值。方法　以MUC1cDNA的引物建立了RT PCR扩增体系及PAGE 银染检测PCR扩增产物的方法 ,并作了优化 ;在对扩增体系的检测特异性和敏感性作了分析后 ,初步检测了临床样品。结果　该扩增体系具有较好的特异性 ,敏感性可达 10 - 6 μgRNA ,相当于从 10 5个淋巴细胞中检出 1个胃癌细胞 ;对临床样品检测的结果与病理检查结果相符。结论　该法具有较好的可靠性 ,可作为临床常规病理探查的补充检查手段。[英文文摘]Objective To develop a RT- PCR method for detecting gastric cancer micrometastases in axillary lymph nodes. Methods T he RT- PCR was set up fo r amplifying MUC1 mRNA and the PAGE- silver staining for detecting amplification products. After optmizing t he PCR system, the spec-i ficity and sensitivity were investigated by amplifying different selected samples and ser ial dilution method respectively . This method was performed on axillary lymph nodes and compared t he results wit h that of patholo gical examination for estimating t he reliability. Results This system showed a good specificity and a high sensitivity of detecting one gastric cancer cell out of 105 lymphocytes. The r esults corresponded with the pathological examination suggesting a good reliability of this method. Conclusion The RT- PCR method possesses a high reliability and promise in clinical appilcation.福建省卫生厅基金!(96075); 厦门大学肿瘤细胞工程开放实验室基金资助项

Challenge by Head Transplant

范瑞平：诸位好!受《中国医学伦理学》杂志王明旭主编所托,组织一篇＂换头术的挑战＂争鸣笔谈,特邀各位参与。请踊跃发表意见,观点不拘,长短不限,畅所欲言,各抒己见,若能针对已发观点形成争论,则更能增加读者兴趣