反油酸对人神经母细胞瘤细胞系SH-SY5Y细胞线粒体功能和细胞自噬的影响

目的研究反油酸对人神经母细胞瘤细胞系SH-SY5Y细胞线粒体功能和细胞自噬的影响。方法体外培养人神经母细胞瘤细胞系SH-SY5Y细胞,加入不同剂量(0、10、20、50和100μmol/L)的反油酸,24 h后收集细胞,采用透射电镜观察细胞自噬小体;采用细胞计数试剂盒8(CCK-8)检测细胞活力;采用实时荧光定量聚合酶链式反应(PCR)和蛋白免疫印迹(western blot)方法检测细胞自噬相关的基因和蛋白表达水平。结果透射电镜观察细胞结构发现,50和100μmol/L反油酸可使细胞内出现自噬小体,并可以引起细胞活力下降,差异均有统计学意义(P0.01);100μmol/L反油酸作用24和48 h后,细胞活力明显降低,差异均有统计学意义(P0.05);100μmol/L反油酸可使SH-SY5Y细胞中细胞色素C(cytochrome c,cyt-c)基因mRNA表达水平明显降低,差异有统计学意义(P0.05);随着反油酸浓度的升高,细胞浆中cyt-c蛋白表达水平逐渐升高,差异均有统计学意义(P0.05)。但是不同剂量的反油酸对SH-SY5Y细胞自噬微管相关蛋白轻链3B(LC3B)基因、Beclin1基因、p62基因、自噬相关基因5(atg5)、自噬相关基因12(atg12)mRNA水平表达差异无统计学意义(P0.05);但与对照组比较,50和100μmol/L反油酸可使SH-SY5Y细胞LC3B和Beclin1蛋白表达水平逐渐上调,差异均有统计学意义(P0.05)。结论反油酸可以引起人神经母细胞瘤细胞系SH-SY5Y细胞活力下降,可能与其引起细胞自噬小体形成,上调自噬相关蛋白表达水平,促进线粒体cyt-c蛋白释放入细胞浆有关。     

Randomised trial investigating effect of a novel nicotine delivery device (Eclipse) and a nicotine oral inhaler on smoking behaviour, nicotine and carbon monoxide exposure, and motivation to quit

OBJECTIVE: To monitor the effect of a novel nicotine delivery device that may produce fewer carcinogens (Eclipse) on cigarette smoking, carbon monoxide and nicotine concentrations, and motivation to give up smoking. The smoker's own brand of cigarette and a nicotine replacement product (Nicotrol inhaler) were used as comparisons. DESIGN: After baseline data were recorded, smokers were randomised to either Eclipse or inhaler for two weeks and then switched to the other product for another two weeks. Thereafter a second baseline was obtained. SETTING AND PARTICIPANTS: Fifty smokers were included and data are reported for the 40 with complete data sets. The smokers were not trying to quit but were interested in trying a new product to reduce their risk. They visited a smoking clinic 10 times during the six week period of the trial. INTERVENTION: No counselling to aid reduction by Eclipse or inhaler was given. MAIN OUTCOME MEASURES: At each visit smoking status and carbon monoxide concentrations were recorded. In half of the visits withdrawal symptoms, attitudes towards smoking, heart rate, and blood nicotine concentrations were also recorded. RESULTS: Eclipse use decreased the number of cigarettes smoked per day (cpd) from 19.1 cpd at baseline to 2.1 cpd (p < 0.001), but increased carbon monoxide concentrations in parts per million (ppm) from 21.0 ppm to 33.0 ppm (p < 0.001). A similar decrease in cigarettes smoked per day was seen with the Nicotrol inhaler, from 19.1 cpd to 4.8 cpd (p < 0.001), but carbon monoxide decreased from 21.0 ppm to 12.7 ppm (p < 0.001). The blood nicotine concentration remained fairly stable with Eclipse, increasing slightly from 16.8 ng/ml to 18.0 ng/ml, while for the inhaler a significant drop was noted, from 16.8 ng/ml to 12. 2 ng/ml (p < 0.002). Craving and withdrawal did not increase with Eclipse. Few significant adverse events occurred with Eclipse. CONCLUSIONS: Eclipse can dramatically decrease cigarette consumption without causing withdrawal symptoms or decreases in nicotine concentrations or motivation to quit altogether. Unlike the inhaler, Eclipse produces an increase in carbon monoxide concentration. Thus Eclipse may not be a safer cigarette.     

姜黄素对乳腺癌MCF-7细胞的抗肿瘤作用及其分子机制

在本实验中,通过MTT法和LDH法检测姜黄素对MCF-7细胞抗肿瘤和毒性作用,通过流式细胞仪的分析揭示了姜黄素对细胞周期、细胞凋亡、线粒体膜电位（△ψm）的影响,利用caspase试剂盒来检验caspase的活性以及用western blot法检测凋亡蛋白Bax、Bcl-2、PARP、细胞色素c的表达。结果显示姜黄素对MCF-7细胞有抗肿瘤以及毒性作用;姜黄素能使MCF-7阻滞在G0/G1期,可以诱导细胞凋亡。在姜黄素作用下细胞的线粒体膜电位（△ψm）增加,caspase-3、caspase-9、Bax、PARP蛋白表达上调,同时细胞色素c从线粒体释放到细胞液中,而Bcl-2的表达减少。这些结果表示姜黄素可以通过使MCF-7细胞的周期阻滞在G0/G1期和通过线粒体途径诱导的凋亡两方面来引起对乳腺癌MCF-7细胞的抗肿瘤作用。      

碳酸氢钾和氯化钠对烟草转化株烟碱转化的诱导作用

以白肋烟TN90高转化株为材料研究了无机盐处理烟株早期叶片对烟碱转化的诱导作用及机理。结果表明,1.2%的碳酸氢钾水溶液和1.2%的氯化钠水溶液均具有显著的诱导转化株早期叶片烟碱转化作用,碳酸钠效果较差。比较碳酸氢钾处理、氯化钠处理分别对常规TN90品种烟株转化株的早期鉴定结果和自然晾制后烟叶的烟碱转化率发现,采用2种无机盐鉴别出的转化株均包括了所有在自然晾制后产生的转化株,且还可鉴别出一些在正常晾制条件下不能有效鉴别的低转化株;早期测定的烟碱转化率一般高于烟叶自然晾制后的烟碱转化率,有利于提高鉴定的准确性和有效性。碳酸氢钾可显著促进叶片内源乙烯的释放,在处理溶液中添加10-4mol/LAVG抑制乙烯的合成可逆转碳酸氢钾对烟碱转化的诱导。     

Grafting tomato plant on tobacco plant and its effect on tomato plant yield and nicotine content

BACKGROUND: Two different tomato scions, cv. Elazig and cv. Sweet (cherry) (Solanum lycopersicum L.) were self‐grafted and grafted onto tobacco root stock (Nicotiana tobacum L.). Then, grafted tomato plants were evaluated in terms of flower and fruit yield. Tobacco‐grafted tomato plant fruits were also evaluated for nicotine content. RESULTS: Tobacco grafting had a positive effect on the tomato plant cultivation performance; the onset of flowering was almost 15 days earlier and the tomato flower and fruit yields increased in both tomato cultivars. Tobacco grafting resulted in 5.0% and 30.1% increase in total fruit weight for cv. Sweet and cv. Elazig, respectively. Because the level of nicotine was within acceptable ranges, tobacco‐grafted tomato fruits were considered to be safe for consumption. Self‐grafted tomato cultivars also had flowering time onsets almost 11 days earlier. However, self‐grafting caused 6.0% and 7.6% less total fruit yield per cv. Sweet and cv. Elazig, respectively. CONCLUSION: In conclusion, our results show that tomato–tobacco grafting is a novel and promising technique for improvement of not only tomato plant performance and yield, but also that it can be employed to various tomato varieties. Copyright © 2009 Society of Chemical Industry     

烤烟烟碱合成及农艺调节效应研究进展

从烟碱合成的部位、前体物质、信号物质以及烟碱在器官间、叶位间和叶片中的分布阐述了烤烟烟碱合成和分布的规律,并对氮、磷、钾等矿质元素,品种、打顶留叶、密度、地膜覆盖、灌溉、成熟度、外源调节物质、环切和断根等农艺措施以及土壤、气候、海拔等环境因子对烤烟烟碱合成的影响进行了综述.     

烤烟烟碱含量与其他化学成分的相关关系及其阈值的研究

本文采用数理统计的方法,系统分析了烤烟烟叶中烟碱含量与总氮、总糖的相关关系。研究表明烤烟烟叶中烟碱含量存在明显的阈值;以烟碱阈值为界,烤烟烟叶中烟碱含量与总氮、总糖的含量存在两种相反的相关关系,当烟碱含量大于阈值时,烟碱与总氮呈正相关关系,与总糖呈负相关关系,当烟碱含量小于阈值时,烟碱与总氮呈负相关关系,与总糖呈正相关关系。      

烟碱抑制单胺氧化酶B活性及其对抗帕金森病作用研究

[背景和目的]流行病学调查结果表明,吸烟与帕金森发病率呈负相关,但吸烟对抗帕金森病的机制仍不清楚.本文拟探讨烟碱对单胺氧化酶B (MAO-B)的抑制作用,明确对抗帕金森病的机制.[方法]通过小分子与蛋白对接模型模拟技术,分析烟碱和MAO-B相互作用及蛋白活性抑制率;对细胞及果蝇的帕金森病模型给予烟碱药物干预,利用分子生...     

一株新的具有高效降低烟碱含量的短小芽孢杆菌MK21的分离筛选及作用研究

采用以烟碱作为唯一碳氮源的选择性培养基,对烟草表面的细菌进行选择性分离,建立降低烟碱含量菌库,通过富集初筛复筛等试验后,从中筛选出一株具有高效降解烟碱能力的细菌MK21。根据形态学特征和生理生化反应及16SrDNA的鉴定,鉴定其为短小芽孢杆菌。将该菌制成菌剂喷施于50 g烟丝样品上,发酵处理5 d。通过测定烟丝的总糖、还原糖、烟碱、氯、钾含量。结果表明:处理后的烟丝中烟碱含量均有下降,其中最高的达到了26.3%。通过短小芽孢杆菌MK21处理后的烟叶样品,达到刺激性降低、掩盖杂气和改善卷烟吸味的效果。     

烟碱脱氢酶基因的原核表达及酶学特性研究

将含烟碱脱氢酶基因重组质粒pBlueScript SK-ndh用限制性内切酶EcoRI、SpeI双酶切,目的基因再亚克隆到非融合表达质粒载体pET-17B,获得重组子质粒pET-17B-ndh,转化大肠杆菌BL21(DE3)pLYS获得高酶活力表达的工程菌株。即使在不用IPTG诱导情况下,该工程菌也产生较高酶活力的可溶性酶,酶产量比野生菌A. nicotinovorans提高25倍。通过硫酸铵沉淀、离子交换层析、凝胶过滤提纯了烟碱脱氢酶,酶被纯化了39.82倍,SDS-PAGE分析得到电泳纯单一条带。该酶活力的最适温度为40℃,该酶组分在pH5.5~8.0范围内稳定,最适pH7.0,对烟碱作用的Km值为7.694×10-4mol/L。Mn2+、Co2+是酶的激活剂,Cu2+是酶的抑制剂。烟碱降解条件研究表明在反应体系中需要辅酶或电子接受体参与介导质子(H+)的转移,该酶催化反应是一种氧化还原反应体系。      

外源乙烯对桃果实采后贮藏过程中线粒体内细胞色素氧化酶的影响

以硬溶质型桃果实‘加纳岩’为试材,研究外源乙烯处理对桃果实采后贮藏过程中呼吸速率、乙烯释放量、细胞色素氧化酶（cytochrome C oxidase,COX）活性以及线粒体编码的COX 3 种亚基（COXⅠ 、COXⅡ 、COXⅢ ）基因表达量的影响。结果表明：外源乙烯处理加速了硬溶质型桃果实‘加纳岩’在采后贮藏过程中硬度的下降,促进了呼吸作用和内源乙烯释放,使呼吸高峰和乙烯释放高峰提前出现。同时外源乙烯抑制了COX酶活性,抑制了贮藏后期COXⅠ 和COXⅡ 亚基基因的表达,抑制了整个贮藏过程中COXⅢ 亚基基因的表达。外源乙烯处理抑制了COX途径的呼吸,加速了果实的软化衰老。