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含非甲基化胞苷-磷酸盐-鸟苷寡脱氧核苷酸在非小细胞肺癌治疗中的应用进展 被引量:2
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作者 胡炜科 任涛 《药学服务与研究》 CAS CSCD 2013年第1期30-33,共4页
非甲基化胞苷-磷酸盐-鸟苷(CpG)寡脱氧核苷酸(ODN)属于Toll样受体9(TLR9)激动剂,其功能主要通过TLR9信号系统来体现。根据构效关系,共鉴别出K型、D型、C型和P型4个类别的CpG ODN,分别对B细胞和浆细胞样树状突细胞产生不同刺激作用。无... 非甲基化胞苷-磷酸盐-鸟苷(CpG)寡脱氧核苷酸(ODN)属于Toll样受体9(TLR9)激动剂,其功能主要通过TLR9信号系统来体现。根据构效关系,共鉴别出K型、D型、C型和P型4个类别的CpG ODN,分别对B细胞和浆细胞样树状突细胞产生不同刺激作用。无论单独应用CpG ODN,抑或联合化疗、放疗,还是联合靶向药物或疫苗,对非小细胞肺癌的治疗均有一定的增效作用。该文也简述了CpG ODN的安全性。 展开更多
关键词 非甲基化胞苷-磷酸-寡脱氧核 非小细 Toll样受体9 肿瘤治疗方案
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5’-磷酸胞苷体外抗氧化作用的研究 被引量:1
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作者 钱佳 施用晖 +1 位作者 何倩 乐国伟 《营养学报》 CAS CSCD 北大核心 2008年第3期316-317,320,共3页
近年来的研究表明,5'-核苷酸具有提高免疫力、促生长、抗氧化、促进机体损伤修复、减少细胞凋亡、减轻炎性反应等功能。但关于5’-核苷酸的抗氧化作用机制尚无系统研究。本研究分析体外条件下5’-磷酸胞苷(5’-CMP)的清除活性氧能... 近年来的研究表明,5'-核苷酸具有提高免疫力、促生长、抗氧化、促进机体损伤修复、减少细胞凋亡、减轻炎性反应等功能。但关于5’-核苷酸的抗氧化作用机制尚无系统研究。本研究分析体外条件下5’-磷酸胞苷(5’-CMP)的清除活性氧能力及其对氧化损伤小鼠脾细胞的修复作用,旨在阐明其可能的作用机制。 展开更多
关键词 5’-磷酸(5’-CMP) 抗氧化作用 自由基 培养
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胞苷环3’,5’一磷酸对正常人淋巴细胞分泌IL-2和IL-2受体表达的抑制作用
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作者 汪明春 李扬秋 +1 位作者 廖继东 姚敏 《中国病理生理杂志》 CAS CSCD 北大核心 1991年第5期473-476,共4页
0.2mM 3’,5’-cCMP不仅能抑制正常人外周血淋巴细胞IL-2的诱生(P<0.01),而且能抑制IL-2受体的表达(P<0.01)。对后者的抑制率为28.9±1.9%。但是在充分IL-2存在的条件下,0.00625mM至0.2mM的3’,5’-cCMP对CTLL-2细胞的增殖有... 0.2mM 3’,5’-cCMP不仅能抑制正常人外周血淋巴细胞IL-2的诱生(P<0.01),而且能抑制IL-2受体的表达(P<0.01)。对后者的抑制率为28.9±1.9%。但是在充分IL-2存在的条件下,0.00625mM至0.2mM的3’,5’-cCMP对CTLL-2细胞的增殖有促进作用,这种促进作用在其浓度较低时更为明显。 展开更多
关键词 淋巴细 -磷酸 受体
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胞嘧啶核苷三磷酸的光合合成 被引量:2
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作者 詹谷宇 莫晓燕 席丹 《化学试剂》 CAS CSCD 北大核心 1996年第5期296-298,共3页
研究了用菠菜叶绿体通过光合磷酸化反应将CMP合成为CTP,本方法具有叶绿体来源方便、生产设备简单、反应时间短、转化率高等优点。约280g菠菜叶可将lgCMP合成1.3gCTP,转化率85%左右。合成的粗品经离子交换提... 研究了用菠菜叶绿体通过光合磷酸化反应将CMP合成为CTP,本方法具有叶绿体来源方便、生产设备简单、反应时间短、转化率高等优点。约280g菠菜叶可将lgCMP合成1.3gCTP,转化率85%左右。合成的粗品经离子交换提纯后,成品的纯度为93.4%,对CMP的收率50.2%。 展开更多
关键词 磷酸 胞苷-磷酸 光合磷酸 生化试剂
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雷公藤4-(5'-二磷酸胞苷)-2-C-甲基-D-赤藓醇激酶基因的全长克隆与表达分析 被引量:8
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作者 童宇茹 苏平 +5 位作者 赵瑜君 张萌 王秀娟 胡添源 高伟 黄璐琦 《中国中药杂志》 CAS CSCD 北大核心 2015年第21期4165-4170,共6页
4-(5-焦磷酸胞苷)-2-C-甲基-D-赤藓醇激酶[4-(cytidine-5-diphospho)-2-C-methyl-D-erythritol kinase,CMK]是雷公藤甲素等萜类成分生物合成途径上关键酶之一。根据获得的雷公藤转录组数据中Tw CMK片段设计特异性引物,采用全长PCR... 4-(5-焦磷酸胞苷)-2-C-甲基-D-赤藓醇激酶[4-(cytidine-5-diphospho)-2-C-methyl-D-erythritol kinase,CMK]是雷公藤甲素等萜类成分生物合成途径上关键酶之一。根据获得的雷公藤转录组数据中Tw CMK片段设计特异性引物,采用全长PCR方法克隆Tw CMK全长c DNA,并进行生物信息学分析;采用实时荧光定量PCR(RT-q PCR)检测茉莉酸甲酯(Me JA)诱导雷公藤悬浮细胞后0~72 h Tw CMK基因的表达水平。结果表明,Tw CMK全长c DNA由1 732个核苷酸组成,具有完整的开放阅读框,共编码387个氨基酸,蛋白相对分子质量为42.85 k Da,理论等电点为5.79;Tw CMK基因受Me JA诱导后表达逐渐上升,在24 h时达到最高值。为进一步研究雷公藤甲素等萜类成分次生代谢调控和生物合成奠定了基础。 展开更多
关键词 雷公藤 4-(5-磷酸)-2-C-甲基-D-赤藓醇激酶(CMK) 生物信息学分析 表达分析
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胡黄连2-C-甲基-D-赤藓糖-4-磷酸胞苷转移酶基因的生物信息学分析 被引量:2
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作者 郑玉娟 陈容 周文化 《中草药》 CAS CSCD 北大核心 2014年第15期2218-2223,共6页
目的研究胡黄连Picrorhiza kurrooa 2-C-甲基-D-赤藓糖-4-磷酸胞苷转移酶(MCT)基因特征并预测MCT结构与功能位点。方法以胡黄连MCT基因为研究对象,利用NCBI网站及生物信息学软件对其碱基分布、氨基酸组成、亲疏水性、保守区及二级结构... 目的研究胡黄连Picrorhiza kurrooa 2-C-甲基-D-赤藓糖-4-磷酸胞苷转移酶(MCT)基因特征并预测MCT结构与功能位点。方法以胡黄连MCT基因为研究对象,利用NCBI网站及生物信息学软件对其碱基分布、氨基酸组成、亲疏水性、保守区及二级结构和三级结构进行预测和分析;并对9个物种的MCT基因进行多重比对与进化分析。结果胡黄连MCT基因的mRNA序列长为1 216 bp(GenBank JQ991625.1),编码由399个氨基酸组成的蛋白质,该蛋白质相对分子质量为44 448.5,其中量最高的为Ser(10.0%);整个多肽中疏水性氨基酸占59.5%,平均疏水值为0.050;与丹参Salvia miltiorrhiza MCT氨基酸序列对比同源性最高,达99%。结论胡黄连MCT基因处于稳定状态,编码的蛋白为疏水性蛋白,在进化过程中是相对保守的,获得的保守区段序列信息为其他物种MCT基因的克隆奠定了基础。 展开更多
关键词 2-C-甲基-D-赤藓糖-4-磷酸转移酶 胡黄连 丹参 生物信息学 蛋白质结构
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樟树4-二磷酸胞苷-2-C-甲基-D-赤藓醇激酶基因的克隆及表达分析 被引量:5
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作者 荆礼 郑汉 +3 位作者 姚娜 陈美兰 申业 黄璐琦 《中国中药杂志》 CAS CSCD 北大核心 2016年第9期1578-1584,共7页
4-二磷酸胞苷-2-C-甲基-D-赤藓醇激酶(4-diphosphocytidyl-2-C-methyl-D-erythritol kinase,CMK)是植物萜类化合物生物合成途径甲基赤藓醇-4-磷酸途径(methylerythritol-4-phosphate pathway,DXP/MEP)上的第4个关键酶。该研究根据樟树... 4-二磷酸胞苷-2-C-甲基-D-赤藓醇激酶(4-diphosphocytidyl-2-C-methyl-D-erythritol kinase,CMK)是植物萜类化合物生物合成途径甲基赤藓醇-4-磷酸途径(methylerythritol-4-phosphate pathway,DXP/MEP)上的第4个关键酶。该研究根据樟树转录组数据,利用RT-PCR(reverse transcription-PCR)方法从药用植物樟树Cinnamomum camphora中克隆得到4-二磷酸胞苷-2-C-甲基-D-赤藓醇激酶基因,命名为Cc CMK1(Gene Bank登录号Ku376098),对其序列进行生物信息学分析,结果表明该基因开放阅读框(ORF)为1 212 bp,编码403个氨基酸,推测其相对分子质量为44.46 k Da,等电点(theoretical p I)为4.99。跨膜结构分析表明CMK蛋白不存在跨膜结构。信号肽分析表明其为非分泌蛋白,不存在信号肽。亚细胞定位表明其定位于叶绿体中。利用荧光实时PCR检测了龙脑樟、油樟、芳樟、脑樟和异樟5种化学型樟树中Cc CMK1基因的表达量,结果表明Cc CMK1基因在油樟中的表达量最高,龙脑樟中表达量最低,为樟树萜类化合物生物合成途径关键酶基因元件挖掘提供研究基础。 展开更多
关键词 樟树 4-磷酸-2-C-甲基-D-赤藓醇激酶(CMK) 基因克隆 生物信息学分析 表达分析
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X-ray Examination of the Complex Adenosine, Guanosine and Cytidine with UO2^2+ Ions
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作者 Aibassov Yerkin Zhakenovich Yemelyanova Valentina +6 位作者 Shakieva Tatyana Nakisbekov Narymzhan Tussupbaev Nessipbay Abenov Bakhyt Bulenbayev Maxat Dossumova Binara Blagikh Evgeniy 《Journal of Chemistry and Chemical Engineering》 2015年第6期408-414,共7页
Interaction of dioxouranitm (VI) UO2^2+ ion with Adenosine-5'-triphosphate, Guanosine-5'-triphosphate and 3ytidine-5'-triphosphate were obtained a complexs of Adenosine, Guanosine and Cytidine with uranium UO2^2... Interaction of dioxouranitm (VI) UO2^2+ ion with Adenosine-5'-triphosphate, Guanosine-5'-triphosphate and 3ytidine-5'-triphosphate were obtained a complexs of Adenosine, Guanosine and Cytidine with uranium UO2^2+ ions and X-ray nethod to explore these complexes. 展开更多
关键词 DNA uranyl ion X-RAY ADENOSINE GUANOSINE Cytidine complexs.
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ATP-bioluminescence assay indicates the treatment of recurrent non-small cell lung cancer combined with malignant pleural effusion
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作者 Jing Zhu Lixia Ma Ying Cheng 《The Chinese-German Journal of Clinical Oncology》 CAS 2010年第8期441-443,共3页
Objective:This study was aimed to research the feasibility of ATP-bioluminescence assay(ATP-TCA) guiding the treatment on recurrent non-small cell lung cancer(NSCLC) combined with malignant pleural effusion.Methods:We... Objective:This study was aimed to research the feasibility of ATP-bioluminescence assay(ATP-TCA) guiding the treatment on recurrent non-small cell lung cancer(NSCLC) combined with malignant pleural effusion.Methods:We collected 30 pleural fluid samples which were approved to be positive by cytology from recurrent NSCLC patients.These cells were cocultured with chemotherapy medicines,single agent or drugs combination.Five drug concentrations,two parallel holes were examined in vitro for 4 days,the results were measured by adding luciferase-fluorescein working system and luminescence analyzer.We applied chemotherapy medicines according to the results in vitro of ATP-TCA.Results:There were differences among drug sensitivities of individuals.All the samples could be evaluated.Effective single drugs included cisplatinum,mitomycin C,doxorubicin,and pemetrexed disodium;sensitive drugs in the combination therapy were gemcitabine plus cisplatin,vinorelbine plus cisplatin,paclitaxel plus cisplatin,docetaxel plus cisplatin,and mitomycin C,vindesine plus cisplatin,in which gemcitabine + cisplatin(GEM + DDP) in vitro was the most efficient program.Conclusion:ATP-TCA in vitro sensitivity assay is rapid,reliable,and simple to guide the treatment of recurrent NSCLC with malignant pleural effusion,and can help clinicians to make the individual chemotherapy program. 展开更多
关键词 ATP-bioluminescence assay (ATP-TCA) recurrent advanced non-small cell lung cancer (NSCLC) pleural fluid
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CRPPA基因变异导致肢带型肌营养不良症1例
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作者 杨媛媛 刘蕊 +7 位作者 范宽 韩璐 刘嘉鹏 李世容 瞿浩 王添阳 刘坤雪灵 胡晓 《中国神经精神疾病杂志》 CAS CSCD 北大核心 2023年第7期431-435,共5页
报告1例CRPPA基因变异所致常染色体隐性遗传性肢带型肌营养不良症患者,总结CRPPA基因相关肌病的临床特点和致病变异谱。患者主要临床表现为双下肢无力7年,进行性加重。血清肌酸激酶水平轻度增高。肌电图提示肌源性损害。肌肉MRI表现为... 报告1例CRPPA基因变异所致常染色体隐性遗传性肢带型肌营养不良症患者,总结CRPPA基因相关肌病的临床特点和致病变异谱。患者主要临床表现为双下肢无力7年,进行性加重。血清肌酸激酶水平轻度增高。肌电图提示肌源性损害。肌肉MRI表现为双侧大腿、小腿多发肌群信号异常。骨骼肌活检组织电镜下可见肌细胞胞浆内肌浆网轻度扩张,小脂滴轻度增多,可见少量萎缩肌细胞。基因检测提示携带CRPPA基因c.1114_1116delGTT(p.V372del)纯合变异。患者父母携带该位点杂合变异。根据ACMG指南将该变异分类为致病性变异。患者明确诊断为肌营养不良-抗肌萎缩相关糖蛋白病C7型。本文拓展了CRPPA基因和抗肌萎缩相关糖蛋白病的遗传变异谱和临床表型谱。 展开更多
关键词 CRPPA基因 肢带型肌营养不良症 遗传性肌肉疾病 抗肌萎缩相关糖蛋白病 磷酸-核糖醇
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磷酸胆碱胞苷酰转移酶α在口腔鳞状细胞癌中的表达及潜在临床意义 被引量:2
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作者 路法超 谢小燕 +1 位作者 尹晓敏 高义军 《中华口腔医学杂志》 CAS CSCD 北大核心 2018年第4期254-258,共5页
目的探讨磷酸胆碱胞苷酰转移酶α(cytidine triphosphate:phosphocholine cytidylyltransferase-α,CCT-α)在口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)中的表达及潜在临床意义。 方法以2016年5月至2016年7月在中南... 目的探讨磷酸胆碱胞苷酰转移酶α(cytidine triphosphate:phosphocholine cytidylyltransferase-α,CCT-α)在口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)中的表达及潜在临床意义。 方法以2016年5月至2016年7月在中南大学湘雅二医院口腔中心收集的58例OSCC组织标本为实验组,同一患者的癌旁组织为对照组。免疫组织化学法检测CCT-α在OSCC组织及癌旁组织中的表达情况,结合患者的临床病理资料分析CCT-α与OSCC患者肿瘤的临床病理特征的关系。采用实时荧光定量PCR及蛋白质印迹法检测OSCC细胞株和正常口腔上皮细胞株中CCT-α mRNA及CCT-α蛋白的表达情况。 结果免疫组化染色结果示CCT-α阳性染色位于细胞核,CCT-α在OSCC组中的表达显著高于癌旁组(P=0.000)。临床病理资料分析示OSCC组织中CCT-α的表达水平与患者吸烟、饮酒史(P=0.001,P=0.004)及肿瘤大小(P=0.005)、分化程度(P=0.000)、是否发生淋巴结转移(P=0.000)密切相关。有吸烟、饮酒史的患者,CCT-α蛋白的表达水平明显升高。实时荧光定量PCR结果显示CCT-α mRNA在OSCC细胞株中的表达显著高于口腔上皮细胞株(P=0.016)。蛋白质印迹法结果显示CCT-α蛋白在OSCC细胞株中的表达显著高于口腔上皮细胞株,其蛋白表达趋势与CCT-α mRNA一致。 结论CCT-α可能在OSCC的发生发展中有重要作用。 展开更多
关键词 口腔鳞状细 口腔肿瘤 磷酸胆碱酰转移酶-α
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滇龙胆GrCMS基因的克隆与表达分析 被引量:2
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作者 张晓东 李彩霞 王元忠 《植物研究》 CAS CSCD 北大核心 2016年第2期258-265,共8页
2-C-甲基-D-赤藓醇4-磷酸胞苷酰转移酶是赤藓糖磷酸酯(MEP)途径中的第三个催化酶。以滇龙胆转录组为基础,采用RT-PCR技术从滇龙胆幼叶克隆2-C-甲基-D-赤藓醇4-磷酸胞苷酰转移酶基因GrCMS,并进行原核表达和组织特异性表达分析。序列分析... 2-C-甲基-D-赤藓醇4-磷酸胞苷酰转移酶是赤藓糖磷酸酯(MEP)途径中的第三个催化酶。以滇龙胆转录组为基础,采用RT-PCR技术从滇龙胆幼叶克隆2-C-甲基-D-赤藓醇4-磷酸胞苷酰转移酶基因GrCMS,并进行原核表达和组织特异性表达分析。序列分析显示,GrCMS基因(登录号KJ917164)开放阅读框(ORF)长933 bp,编码310氨基酸,推测分子量为34.23 kD,等电点为7.68。蛋白质序列分析显示,GrCMS无信号肽,为亲水稳定蛋白,主要由α-螺旋和无规则卷曲构成,可能定位叶绿体;具有CMS保守结构域。进化分析结果表明,GrCMS蛋白与长春花CrCMS蛋白亲缘关系最近。原核表达结果表明,GrCMS与预期蛋白大小一致。定量PCR结果表明,GrCMS基因主要在叶中表达。结果为进一步研究该基因功能和龙胆苦苷生物合成途径奠定基础。 展开更多
关键词 滇龙胆 2-C-甲基-D-赤藓醇4-磷酸酰转移酶 基因克隆 表达分析
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OSMOTIC STRESS DECREASES THE ACTIVITY OF ATPASE ASSOCIATED WITH THE ROOT CAP PLASMODESMATAIN ZEA MAYS 被引量:1
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作者 苗龙 王学臣 张伟成 《Acta Botanica Sinica》 CSCD 1997年第8期774-777,共4页
With light and electron microscopy the substructural change and the ATPase activity of corn (Zea mays L.) root cap cells after short-term osmotic stress were studied. Some spoke-like fine strands originating from the ... With light and electron microscopy the substructural change and the ATPase activity of corn (Zea mays L.) root cap cells after short-term osmotic stress were studied. Some spoke-like fine strands originating from the departed periplasm and stretching towards cell wall could be observed even after plasmolysis. By observing the precipitation of ATPase activity product (lead phosphate) at plasma membrane and plasmodesmata, it was found that the fine strands were plasma membrane-lined channels surrounding the cytoplasm and that they still firmly connected to the plasmodesmata during plasmolysis. Compared with the control (unstressed), a sharp decrease of ATPase activity in the plasmodesmata of the stressed cells was observed. Inhibition of energy metabolism in these limited locales would affect the physiological activity, maybe including the regulation of permeability and the change of size exclusion limit (SEL) of plasmodesmata. 展开更多
关键词 ATPASE Osmotic stress PLASMODESMATA Zea mays
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不同强化剂对柯乐猪CMAH和GGTA1基因表达的影响 被引量:1
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作者 杨仕钰 燕志宏 +2 位作者 刘东云 吴开慧 张芸 《中国畜牧杂志》 CAS 北大核心 2022年第3期161-165,共5页
为提升地方猪猪肉品质,开发高端猪肉食品,本实验研究了添加5’-胞苷单磷酸(5’-CMP)、山奈酚(KA)、单宁酸(TA)以及混合强化剂对柯乐猪肝脏和背最长肌CMAH和GGTA1基因表达的影响。实验选取体重相近、日龄相当和免疫水平等外部因素均相同... 为提升地方猪猪肉品质,开发高端猪肉食品,本实验研究了添加5’-胞苷单磷酸(5’-CMP)、山奈酚(KA)、单宁酸(TA)以及混合强化剂对柯乐猪肝脏和背最长肌CMAH和GGTA1基因表达的影响。实验选取体重相近、日龄相当和免疫水平等外部因素均相同的50头去势柯乐猪,并将其随机分为5组,饲喂不同强化剂60 d,饲喂后随机选择3头进行屠宰,收集肝脏和背最长肌,提取总RNA,并使用实时荧光定量PCR检测CMAH和GGTA1在柯乐猪肝脏和背最长肌中的实际表达量。结果表明:饲料中添加5’-CMP和KA能抑制肝脏和背最长肌CMAH基因表达,添加TA和含有TA的混合组对肝脏CMAH基因抑制效果不显著,但在背最长肌中显著高于KA组和5’-CMP组;TA组和混合组肝脏GGTA1基因表达量差异极显著高于对照组、5’-CMP组、KA组,对照组中GGTA1基因表达量与5’-CMP组和KA组差异不显著,混合组和对照组背最长肌GGTA1基因表达量显著高于5’-CMP组、KA组,与TA组差异不显著。本次研究为开发高端猪肉市场和器官移植、红肉过敏症等研究提供了基础数据。 展开更多
关键词 CMAH GGTA1 5’-磷酸 山奈酚 单宁酸
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Induction of nucleoside phosphorylase in Enterobacter aerogenes and enzymatic synthesis of adenine arabinoside 被引量:5
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作者 Xiao-kun WEI Qing-bao DING +3 位作者 Lu ZHANG Yong-li GUO Lin OU Chang-lu WANG 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2008年第7期520-526,共7页
Nucleoside phosphorylases (NPases) were found to be induced in Enterobacter aerogenes DGO-04, and cytidine and cytidine 5′-monophosphate (CMP) were the best inducers. Five mmol/L to fifteen mmol/L cytidine or CMP cou... Nucleoside phosphorylases (NPases) were found to be induced in Enterobacter aerogenes DGO-04, and cytidine and cytidine 5′-monophosphate (CMP) were the best inducers. Five mmol/L to fifteen mmol/L cytidine or CMP could distinctly increase the activities of purine nucleoside phosphorylase (PNPase), uridine phosphorylase (UPase) and thymidine phosphorylase (TPase) when they were added into medium from 0 to 8 h. In the process of enzymatic synthesis of adenine arabinoside from adenine and uracil arabinoside with wet cells of Enterobacter aerogenes DGO-04 induced by cytidine or CMP, the reaction time could be shortened from 36 to 6 h. After enzymatic reaction the activity of NPase in the cells induced remained higher than that in the cells uninduced. 展开更多
关键词 Nucleoside phosphorylase (NPase) Enterobacter aerogenes CYTIDINE Cytidine 5'-monophosphate (CMP) Adenine arabinoside
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Co regulative effects of the cAMP/PKA and DAG/PKC signal pathways on human gastric cancer cells during differentiation induced by traditional Chinese medicines *
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作者 谷善青 梁云燕 +2 位作者 樊立人 李宝元 王代树 《World Journal of Gastroenterology》 SCIE CAS CSCD 1997年第1期56+53-55,53-55,共4页
AIM To evaluate the role of cAMP/PKA and DAG/PKC pathways of the MGc80 3 cells treated with traditional Chinese medicine of compound Bailong preparation (Bailong).
关键词 Chinese medicine cAMP/PKA DAG/PKC Gastric cancer
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Inhibitory effects of extracellular adenosine triphosphate on growth of esophageal carcinoma cells 被引量:4
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作者 Ming-Xia Wang Lei-Ming Ren Bao-En Shan 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第38期5915-5919,共5页
AIM: To study the growth inhibitory effects of ATP on TE-13 human squamous esophageal carcinoma cellsin vitro.METHODS: NTT assay was used to determine the inhibition of proliferation of ATP or adenosine (ADO) on T... AIM: To study the growth inhibitory effects of ATP on TE-13 human squamous esophageal carcinoma cellsin vitro.METHODS: NTT assay was used to determine the inhibition of proliferation of ATP or adenosine (ADO) on TE-13 cell line. The morphological changes of TE-13 cells induced by ATP or ADO were observed under fluorescence light microscope by acridine orange (AO)/ethidium bromide (EB) double stained cells. The intemudeosomal fragmentation of genomic DNA was detected by agarose gel electrophoresis. The apoptotic rate and cell cycle after treatment with ATP or ADO were determined by flow cytometry.RESULTS: ATP and ADO produced inhibitory effects on TE-13 cells at the concentration between 0.01 and 1.0 mmol/L. The ICs0 of TE-13 cells exposed to ATP or ADO for 48 and 72 h was 0.71 or 1.05, and 0.21 or 0.19 mmol/L, respectively. The distribution of cell cycle phase and proliferation index (PI) value of TE-13 cells changed, when being exposed to ATP or ADO at the concentrations of 0.01, 0.1, and 1 mmol/L for 48 h. ATP and ADO inhibited the cell proliferation by changing the distribution of cell cycle phase via either G0/G1 phase (ATP or ADO, 1 mmol/L) or S phase (ATP, 0.1 mmol/L) arrest. Under light microscope, the tumor cells exposed to 0.3 mmol/L ATP or ADO displayed morphological changes of apoptosis. A ladder-like pattern of DNA fragmentation was obtained from TE-13 cells treated with 0.1-1 mmol/L ATP or ADO in agarose gel electrophoresis. ATP and ADO induced apoptosis of TE-13 cells in a dose-dependent manner at the concentration between 0.03 and 1 mmol/L. The maximum apoptotic rate of TE-13 cells exposed to ATP or ADO for 48 h was 16.63% or 16.9%, respectively.CONCLUSION: ATP and ADO inhibit cell proliferation, arrest cell cycle, and induce apoptosis of TE-13 cell line. 展开更多
关键词 Extracellular adenosine triphosphate Esophageal carcinoma cells APOPTOSIS Growth inhibition
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LYSOPHOSPHATIDIC ACID ACTIVATES NUCLEAR NUCLEOSIDE TRIPHOSPHATASE IN RAT HEPATOCYTES
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作者 李菊香 李载权 +2 位作者 伍期专 唐朝枢 杜军保 《Chinese Medical Sciences Journal》 CAS CSCD 2003年第4期213-217,共5页
Objectives.To observe if lysophosphatidic acid(LPA)can influence nuclear nucleoside triphos-phatase(NTPase)activity of isolated hepatocyte from rat,and to investigate the possible mechanisms by which LPA affects the N... Objectives.To observe if lysophosphatidic acid(LPA)can influence nuclear nucleoside triphos-phatase(NTPase)activity of isolated hepatocyte from rat,and to investigate the possible mechanisms by which LPA affects the NTPase.Method.Isolated and cultured hepatocytes from rat liver were exposed to LPA(1×10 -9 ,1×10 -8 and5×10 -8 mol/L)with or without inhibitors of protein kinase C(PKC)and mitogen activating protein kinase kinase(MAPKK),and the NTPase activity on nuclear envelope was assayed using ATP and GTP as substrate,respectively.Results.Nuclear NTPase activity of rat hepatocytes was potently stimulated by incubation of hepato-cytes with LPA in concentration?and time ?dependent manners.In hepatocytes incubated with LPA,nu-clear NTPase activity was significantly higher than that of the control(P<0.01).In hepatocytes preincu-bated with PKC inhibitor H-7or MAPKK inhibitor PD98059,LPA-stimulated activation of nuclear NT-Pase was obviously attenuated.In addition,direct incubation of isolated hepatic nuclei with LPA had no effect on nuclear NTPase activity.Conclusion.LPA is involved in modulating nuclear NTPase activity in hepatocytes.The stimulating effect of LPA on the nuclear NTPase is mediated at least partly by PKC and MAPK-dependent pathway. 展开更多
关键词 lysophoshatidic acid nucleoside triphosphatase HEPATOCYTE
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Ultrastructural and Ultrahistochemical Studies on Rat Hepatocytes under the Effect of Lead Acetate Administration
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作者 Awatef Mohamed Ali Wafaa Abdel-Rhaman Ahmad Mona Abdel-Hamed Yehia 《Journal of Life Sciences》 2010年第6期21-28,共8页
The present study was carried out to investigate the effect of lead acetate on the ultrastructure of albino rat hepatocytes with special reference to its effect on the mitochondrial and lysosomal activity. Lead acetat... The present study was carried out to investigate the effect of lead acetate on the ultrastructure of albino rat hepatocytes with special reference to its effect on the mitochondrial and lysosomal activity. Lead acetate was given orally to albino rats at a dose of 1% lead acetate / 100 g body weight, three times / week for one month (Gila), two months (Glib), three months (GIIc). Liver total protein, body / liver weight ratio and cytochrome P-450 value were calculated. Three parts of the liver samples were incubated in media containing adenosine triphosphate, p-nitrophenyl phosphate and 2% glutlraldehyde and prepared for electron microscopy to visualize adenosine triphosphatase, acid phosphatase and the fine structures ofhepatocytes respectively. The main changes of the fine structures was found in the nucleus, as irregularity of the nuclear membrane, clumped heterochromatin and sun radiation of lead inclusion bodies. The cytoplasm was characterized by shortened, dilated rough endoplasmic reticulum, destroyed and hazy mitochondria with increased number of lysosomes with storage secretion. Ultrastructure findings showed hepatocytes damage due to increased hydrolysis enzymes and decreased cytotoxic enzymes (cytochrome P-450) as well as oxidative enzymes that proved the toxic effect of lead according to the duration of exposure. 展开更多
关键词 Acid phosphatase adenosine triphosphatase HEPATOCYTE lead acetate u|trastructure.
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茅苍术AlCMK基因的克隆及原核表达分析 被引量:1
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作者 鲁继梅 徐睿 +4 位作者 吴君贤 邹立思 刘超 彭华胜 查良平 《药学学报》 CAS CSCD 北大核心 2022年第9期2876-2884,共9页
4-二磷酸胞苷-2-C-甲基-D-赤藓醇激酶[4-(cytidine-5′-diphospho)-2-C-methyl-D-erythritol kinase,CMK]是生成萜类化合物MEP途径的关键酶之一。本研究以茅苍术转录组数据为依据,克隆获得CMK基因序列,命名为AlCMK(GenBank登录号为OM283... 4-二磷酸胞苷-2-C-甲基-D-赤藓醇激酶[4-(cytidine-5′-diphospho)-2-C-methyl-D-erythritol kinase,CMK]是生成萜类化合物MEP途径的关键酶之一。本研究以茅苍术转录组数据为依据,克隆获得CMK基因序列,命名为AlCMK(GenBank登录号为OM283293)。研究结果表明,AlCMK包含一个全长为1230 bp的开放阅读框(ORF),编码409个氨基酸,推测其相对分子质量为44752.53,蛋白理论等电点为6.67;跨膜结构分析表明其无跨膜结构,二级结构显示其主要由无规则卷曲(48.17%)结构组成;同源氨基酸序列分析表明,茅苍术与除虫菊、桂花、杜仲、忍冬、丹参CMK基因编码的氨基酸序列具有较高同源性;系统进化树分析表明,茅苍术AlCMK与菊科植物CMK蛋白亲缘关系较近;构建pET-32a-AlCMK原核表达载体,并转化至大肠杆菌BL21(DE3)表达感受态,蛋白表达结果显示其分子质量约为65 kDa;利用qRT-PCR分析AlCMK基因在不同组织和茉莉酸甲酯(MeJA)处理后的表达模式,同时采用ELISA试剂盒法测定茅苍术CMK酶活性,结果表明,不同产地茅苍术AlCMK基因具有组织表达差异性,且受外源MeJA诱导表达,酶活结果显示不同产地茅苍术CMK酶活性均在叶中较高;亚细胞定位显示该基因位于叶绿体中。本研究为进一步阐明茅苍术中萜类化合物合成途径AlCMK基因的生物学功能提供参考。 展开更多
关键词 茅苍术 4-磷酸-2-C-甲基-D-赤藓醇激酶 基因克隆 原核表达 表达分析 亚细定位
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