目的 建立BRL-3A细胞系胰岛素抵抗模型,筛选棕榈酸诱导的最佳条件,并探究其作用机制.方法 采用CCK8法确定棕榈酸对BRL-3A细胞增殖的影响;采用葡萄糖检测试剂盒检测葡萄糖含量,确定不同浓度的棕榈酸对BRL-3A细胞摄取葡萄糖的影响;采用免疫印迹法(Western blotting)检测胰岛素受体底物1(IRS-1)、磷脂酰肌醇-3'-羟基激酶(PI3K)、磷酸化丝氨酸-苏氨酸激酶(p-AKT)、丝氨酸-苏氨酸激酶(AKT)、葡萄糖转运蛋白4(GLUT4)的蛋白表达.结果 0.2 mmol·L-1棕榈酸作用36 h对BRL-3A细胞增殖无影响;0.4 mmol·L-1棕榈酸诱导6 h、0.25和0.2 mmol·L-1棕榈酸诱导12 h,BRL-3A细胞开始出现明显胰岛素抵抗,移除刺激后48 h内仍然能够抑制胰岛素刺激状态下的糖消耗;在蛋白水平,棕榈酸降低了IRS-1、PI3K、p-AKT、GLUT4在BRL-3A细胞内的表达.结论 高浓度棕榈酸能够通过抑制IRS-1等靶蛋白表达诱导BRL-3A细胞产生胰岛素抵抗.

目的:建立LC-MS/MS法测定达芦那韦在大鼠血浆中的浓度并进行药动学研究.方法:以卡马西平为内标,采用甲醇沉淀蛋白方法处理样品,富集血浆样品中的达芦那韦,用LC-MS/MS进行检测.色谱柱为Inertsil?ODS3 C18柱(150 mm×4.6 mm,5μm),C18保护柱(50 mm×4.6 mm),流动相:甲醇-水溶液(90:10),流速:0.5 ml·min-1,柱温:30℃,采用电喷雾电离源(ESI),正离子模式,多反应监测(MRM)扫描分析,达芦那韦和内标的离子迁移通道分别为m/z 548.5→530.5[碰撞能量(CE)=25 eV,去簇电压(DP)=80 V],m/z 237.1→194.1(CE=18 eV,DP=150 V);大鼠口服给药后,分别于各时间点取大鼠血浆,测定血浆中的药物含量,并用DAS 2.0软件计算其药动学参数.结果:达芦那韦在0.01～10.0μg·ml-1范围内线性良好,提取回收率≥92.13％,日内与日间精密度RSD≤6.6％.大鼠口服达芦那韦后AUC(0-t)为(1416.23±370.13)mg·L-1·min,AUC(0-∞)为(1493.70±374.15)mg·L-1·min,tmax为360.00 min,Cmax为(3.03±0.85)μg·ml-1.结论:该方法方便、快速、精确,能够应用于大鼠血浆中达芦那韦的测定及药动学研究.

晕动病又称运动病,是指机体对异常运动感知的正常应激性生理反应,不论这种感知是真实的还是虚拟的,晕动病的严重程度往往与运动刺激程度和个体差异有关.通过晕动病的动物模型,深入了解眩晕的发病机制,对于眩晕的治疗有重要意义.本文分别从动物眩晕模型的造模方法和评价指标进行综述.

目的:探讨参花凝胶对豚鼠激素依赖性皮炎的作用.方法:将56只健康豚鼠(雌雄各半)随机分为空白对照组、模型组、空白基质组、参花凝胶低(以生药材计1.5％)、中(以生药材计3.0％)、高(以生药材计6.0％)剂量组及阳性药物他克莫司组(0.03％),每组8只.除空白对照组外,其余各组采用0.05％丙酸氯倍他索酊剂建立豚鼠激素依赖性皮炎动物模型.造模成功后,各组大鼠涂抹给药,剂量为使液体均匀覆盖裸露皮肤即可,2次/d,连续15 d.观察实验前后各组豚鼠的皮损和病理改变,测定血清IgE含量变化.结果:与空白对照组比较,模型组皮肤组织切片中乳头层和网织层排列最紊乱,炎症部位最多,并且血清中IgE水平显著升高(P<0.05),说明造模成功.与模型组比较,参花凝胶低、中、高剂量组皮肤外观好转,血清总IgE水平降低,并且参花凝胶高剂量组与模型组比较差异有统计学意义(P<0.05).HE染色和硝酸银染色后镜下观察,与模型组相比,各个给药组皮肤组织均得到一定修复,损伤和炎症减少.结论:参花凝胶对豚鼠激素依赖性皮炎有缓解和治疗作用.

Ex-RAD (ON01210) is a novel and efficient anti-radiation drug with low toxicity developed by US army and Onconova Pharmaceuticals in recent years. The significant survival advantage and low toxicity of Ex-RAD have contributed to the approval by the FDA as an investigational new drug in December 2008. Meanwhile, the drug is currently in phase I clinical trials in humans. In this paper, the chemical structure, synthesis route, detection method and pharmacological action of Ex-RAD were reviewed, and the appli-cation prospect of Ex-RAD was also explored.

Objective To research the influence of Alismatis combined with bifendate on cytochrome P 450 in rat liver microsomes. Methods Twenty four healthy male SD rats were randomly divided into four groups:the experimental groups were given Alismatis at 140 mg·kg-1 , bifendateat 2.18 mg·kg-1 , and Alismatis plus bifendate at 140 mg+2.18 mg·kg-1 ,while the blank control group was given 0. 9% sodium chloride at 5 Ml · kg-1 . The liver microsomes were prepared upon differential centrifugation 7 days after administration.The microsomal protein concentration, cytochrome P450 content, Cytb5 content, NADPH cytochrome C reductase activity and amino pyrine N removal of methyl enzyme activity, erythromycin demethylase activity were determined by UV respectively. Results Compared with the normal control group, the combination use of Alismatis and bifendate redued the microsome content and cytochrome P450 content, while it increased the NADPH cytodrome C activity. The concentrations of cytochrome P450 were both increased by Alismatis and bifendate. Conclusion In contrast, combination ofAlismatis and bifendate reduces cytochrome P450 content which has a negative effect on phase I drug metabolism.Moreover, the combination of Alismatis and bifendate induced NADPH- cytochrome C reductase, accelerated the reduction of cytochrome P450 and inhibited cytochrome P450 isoform CYP2E1 activity.

Objective:To develop an LC-MS/MS method for the quantitative determination of phloretin in plasma of Beagle dogs to study the pharmacokinetics of phloretin in Beagle dogs. Methods:With resveratrol as the internal standard, LC-MS/MS was adopted to determine phloretin in plasma of Beagle dogs after the protein precipitation by ethyl acetate. An Agilent EC C18 column ( 100 mm × 4. 6 mm, 3. 5 μm) was used and the column temperature was 30℃. The mobile phase was acetonitrile-water (95 :5) and the flow rate was 0. 5 ml·min-1 . The injection volume was 5 μl. The samples were detected by tandem mass spectrometry with the electros-pray ionization ( ESI) source in the multiple reaction monitoring ( MRM) mode. The anion-selective channel of phloretin and the inter-nal standard was m/z 273. 0→166. 9 and 227. 0→184. 9, respectively. The concentration of phloretin in plasma was at various time points tested after the oral administration of phloretin capsules at the dosage of 30 mg·kg-1 ,60 mg·kg-1 and 120 mg·kg-1 and the data were processed with DAS. 2. 0 software. Results: The standard calibration curve of phloreitn was linear over the range 50. 0 ~125000. 0 ng·ml-1 . The recoveries obtained for phloreitn were greater than 93. 15%. The intra-day and inter-day RSDs were less than 6. 1%. After the oral administration at the dose of 30,60 and 120 mg·kg-1, tmax was (180. 0 ± 69. 28)min, 240 min and 240 min,and Cmax was (1025. 1 ± 189. 2) ng·ml-1 , (3075. 9 ± 242. 3) ng·ml-1 and (5712. 3 ± 600. 8) ng·ml-1 ,respectively. There was a positive correlation between AUC and administration dosage. Conclusion:The method is rapid, accurate and simple, which is successfully applied in the pharmacokinetic study of phloretin in Beagle dogs. As the dosage increases, AUC and Cmax both increase with stable t1/2 and decreased CL/F.

目的:建立LC-MS/MS法测定卡巴他赛在比格犬血浆中的浓度,并将该方法应用于卡巴他赛注射液在比格犬体内的药动学研究.方法:以卡马西平为内标,血浆样品经甲醇沉淀蛋白,富集生物样品中的卡巴他赛,然后进行LC-MS/MS检测.色谱柱采用Agilent EC C18柱(50 mm×4.6 mm,2.7 μm),柱温为30℃,以甲醇-水(95∶5)为流动相,流速为0.5 ml·min-1,进样量为5μl,采用电喷雾电离源(ESI),正离子多反应监测(MRM)扫描分析,卡巴他赛和内标的离子选择通道分别为m/z858.4→577.4,m/z 237.1→194.1;比格犬单剂量静脉静滴卡巴他赛注射液20 mg·h-1,于不同时间点取血测定血浆中药物的含量并利用DAS 2.0软件计算其药动学参数.结果:卡巴他赛在10.0～1 000.0 ng·ml-1范围内线性关系良好,平均回收率均大于93.53％,日内与日间精密度(RSD)均小于7.4％.比格犬静脉注射卡巴他赛后AUC(0-t)为(155 181.93±11 593.33)ng·ml-1·min-1,AUC(0-∞)为(167 528.12±16671.46) ng·ml-1·min-1,tmax为60 min,Cmax为(688.37±52.06) ng· ml-1.结论:该法快速、精确、简便,可用于比格犬血浆中卡巴他赛的测定及药动学研究.

目的:建立L6细胞胰岛素抵抗模型,筛选出最佳诱导条件,并探讨其作用机制.方法:用MTT法确定棕榈酸、胰岛素对L6细胞增殖的影响;用葡萄糖检测试剂盒检测葡萄糖含量,确定不同浓度的棕榈酸、胰岛素对L6细胞葡萄糖消耗的影响;通过Western Blot法检测IRS-1、PI3K、P-AKT、AKT、GLUT4的蛋白表达.结果:0.4 mmol·L-1棕榈酸诱导12 h、5×10-7mol· L-1胰岛素诱导24 h以上,对L6细胞生长无影响且出现明显胰岛素抵抗,移除刺激后24h内能够抑制胰岛素刺激状态下的糖转运;棕榈酸降低了IRS-1、PI3K、P-AKT、GLUT4在L6细胞上的表达.结论:高浓度棕榈酸能够通过抑制IRS-1等靶蛋白表达诱导L6细胞产生胰岛素抵抗.

Theacrine is one kind of natural purine alkaloids, which mainly exists in an unusual Chinese tea known as Kucha. It shows various biological activities, such as anti-depression, sedative and hypnotic effects and anti-inflammatory and analgesic activi-ties. The study on theacrine dates back to a few decades ago. According to the references published in recent years, the resource, preparation, characterization and pharmacological effects of theacrine were reviewed, and its application prospect was also explored.

目的 探讨人小细胞肺癌多药耐药细胞H446/CDDP细胞内活性氧(ROS)介导的线粒体功能不全及其与顺铂耐药性之间的关系.方法 用MTT法检测H446细胞及其多药耐药细胞H446/CDDP在5μg/ml顺铂作用后的细胞生存率,流式细胞术检测细胞凋亡,荧光显微镜观察顺铂诱导后两种细胞内ROS含量以及线粒体膜电位(MMP)水平.实验中均设置空白对照组.结果 顺铂诱导后细胞的生存率均随药物作用时间的延长明显降低,且H446/CDDP细胞在各个时间点的生存率均明显高于H446细胞;5μg/ml顺铂作用24 h后H446和H446/CDDP两种细胞的早期凋亡百分比分别为(8.63±0.45)％和(1.97±0.42)％,具有明显的统计学差异(P ＜0.05);H446细胞内ROS的水平在顺铂诱导24 h后明显升高,而H446/CDDP细胞内ROS的水平无明显变化;与此同时,H446细胞MMP下降水平明显高于H446/CDDP细胞.结论 顺铂可使小细胞肺癌H446细胞内ROS的产生增加,MMP下降,从而诱导细胞凋亡,而多药耐药细胞H446/CDDP顺铂耐药性的产生与顺铂作用后细胞内ROS减少,MMP下降抑制所致的线粒体介导的内源性细胞凋亡途径耐受有关.

Objective To develop an HPLC-MS/ MS method for quantitative determination of PA-824 in rat plasma and to study the pharmacokinetics of PA-824 in rat after oral administration. Methods An HPLC-MS/ MS method was developed and validated for determination of PA-824 in rat plasma using metronidazole as internal standard.The proteins in plasma samples were precipitated with methanol,and PA-824 was enriched for analysis by HPLC-MS/ MS.An Inertsil? ODS3 C18 column (150 mm×4.6 mm,5 μm) was applied with mobile phase composed of methanol- 0.03% triethylamine (TEA) in water (90:10) ,at a flow rate of 0. 5 mL ? min-1 and column temperature of 30 ℃ . Quantitation was performed on a triple quadrupole mass spectrometer applying electrospray ionization technique and operating in multiple reaction monitoring (MRM) and positive ion mode with transitions at 360.1/ 175.0 for PA-824 and 172.0/ 128.0 for metronidazole.The concentration of PA-824 in plasma was tested after oral administration at various time points and the data were processed with software DAS.2.0. Results The standard calibration curve for spiked rat plasma containing PA-824 was linear over the range of 0. 1 - 10. 0 μg?mL-1 . The recoveries obtained for PA-824 were greater than 92.13%.Intra-day and inter-day coefficient of variation were less than 6.6%.After oral administration,the main pharmacokinetic parameters were AUC(0-t) : ( 3 297. 503 ± 320. 958) mg ? L-1 ? min-1 , AUC(0-∞ ) :(3 558.315±338.860)mg?L-1?min-1 ,tmax:(360.000±64.143)min,Cmax:(3.5±0.3)μg?mL-1 . Conclusion The method is rapid,accurate,simple,and successfully applied in a pharmacokinetic study of fixed dose oral administration of PA-824 in rats.

目的 改进抗肿瘤药物卡巴他赛的合成方法.方法 以10-去乙酰基巴卡亭Ⅲ(10-DABⅢ)为起始原料,经过双甲醚化、酯化偶联、脱保护3步反应制得目标产物.结果与结论 采用不同保护基的C-13侧链对酯化偶联反应具有重要的影响;确定了一条实用的制备卡巴他赛的合成路线,3步反应总收率达到41.7％(以10-DABⅢ计).目标产物的结构经1 H-NMR和MS谱确证.该合成路线适于批量合成.