单纯疱疹病毒对常用抗病毒药物的耐药性

单纯疱疹病毒(HSV)与药物共同培养,传数代后,对单一抗病毒药物无环鸟苷(ACV)、环胞苷(CC)、丙氧鸟苷(DHPG)和膦羧基甲酸钠(PFA)均可产生耐药性。但耐药性产生的时间和程度各有不同。联合用药可延缓并减少病毒耐药株的发生,同时可降低用药浓度和剂量,从而减少毒性反应。不同耐药株的交叉药敏试验结果为临床合理用药提供了实验依据。     

噻芬太尼的主要药效和致依赖性潜力的实验研究

本文研究噻芬太尼的镇痛药效和制动作用,评价其致身体依赖性潜力。小鼠热板法测得噻芬太尼的镇痛作用强度分别为吗啡、芬太尼和埃托啡的3260,22和1.5倍。以瘫痪作为制动指标测得噻芬太尼对大鼠、家兔、狗和猴制动作用强度为埃托啡的2～3倍。小鼠跳跃实验和大鼠饮药液自然戒断实验表明该药具有一定致身体依赖性潜力。大鼠ⅳ快速成瘾实验及长达20周的猴身体依赖性实验则未出现依赖性戒断症状。     

Expression and characterization of Gag protein of HIV-1 （CN） in Pichia pastoris

To express the core protein of HIV-1 of Chinese prevalent strata （HIV-1[CN]） in Pichia pastoris, the fulllength gag gene was inserted into the secretory expression vector pHILS1. Linearized recombinant plasmid pHILGAG by Sal I was electrotransformed into the yeast strain GS115, and the yeast transformants were identified by PCR. To induce the interest protein to be expressed, the PCR positive transformants were inoculated in the medium of BMGY and BMMY, mRNA of the strain was detected by RT-PCR, and the expressed protein was analyzed by SDS-PAGE, Western blotting and thin layer scanning. mRNA （1.3 kb） was amplified by RT-PCR. SDS-PAGE and Western blotting analysis showed that the molecular mass of the expressed protein was 55 kDa, which was similar to the expected value, and the expressed protein could react with McAb to HIV-1 p24. Thin layer scanning analysis demonstrated that the whole amount of the expressed protein was approximately 13% of the soluble protein in the supernatant. The recombinant yeast had good genetic stability. The optimal expression conditions of the engineering yeast were as follows： BMMY medium, 80%-90% of dissolved oxygen, 1% methanol, and 3-day-cultivation course. Gag proteins were expressed under the optimal expression condition and purified via gel filtration chromatography. The purity of the interest protein was up to 85%. After the purified proteins were inoculated into BALB/c mice, the anti-HIV-1 antibodies in the immunized mice could be detected by Western blotting.     

信号转导子与转录激活子3在豚鼠形觉剥夺性近视发展中的表达特征

目的:观察高度近视眼眼球后壁组织信号转导子与转录激活子3的动态表达及意义。方法:实验于2005-10/2006-08在郑州大学基础医学院完成。选用健康1周龄豚鼠30只,雌雄不拘,无眼部疾患,实验前对豚鼠进行检影验光,排除先天性近视。右眼遮盖作为实验组,将直径为15mm的半透明眼罩用502胶粘于豚鼠眼周围的皮肤上,分别以1,3,6周3个不同时间点持续遮盖,每个时间点10只。左眼不做任何处理作为对照组。两组均采用检影验光检测屈光度;眼科A超测定眼轴长度;对两组3个时间点眼球后壁行S-P法免疫组织化学染色,检测信号转导子与转录激活子3的动态表达。结果:纳入豚鼠30只,均进入结果分析,模型制备过程无眼罩脱落。①生后1周豚鼠均呈远视状态,眼轴短;随时间延长,对照组远视度数减低,眼轴延长;实验组由实验前远视眼快速演变为高度近视眼,并随遮盖时间延长,近视度数加深,眼轴延长,两组之间实验前眼屈光度数、眼轴长度无明显差异(P>0.05)。实验组形觉剥夺后,两组之间眼屈光度数、眼轴长度进行比较,差异有非常显著性意义(P<0.01)。②视网膜光感受器外节、色素上皮-脉络膜均有信号转导子与转录激活子3阳性表达、巩膜弱阳性表达;而对照组表达主要位于神经节细胞,随出生时间延长,两组信号转导子与转录激活子3表达均逐渐下调。与对照组相比,实验组信号转导子与转录激活子3表达明显上调,但两组之间比较,自实验第1周开始,实验组信号转导子与转录激活子3较对照组明显上调,差异有显著性意义(P<0.01)。结论:形觉剥夺可导致高度近视;在近视中信号转导子与转录激活子3表达主要位于视网膜光感受器外节、色素上皮层和脉络膜,可能参与了近视眼的形成与发展。     

扶正益气中药对力竭运动后优秀赛艇运动员淋巴细胞的作用

目的:观察以黄芪、灵芝提取物为主要成分的扶正益气中药对大强度力竭运动后优秀赛艇运动员外周血淋巴细胞的影响。方法:于2004-08/10选择国家健将级赛艇男运动员16名,随机数字表法分为对照组和实验组,每组8名。实验组和对照组受试者分别在测试前,每天早、晚各1次服用扶正益气中药(主要成分为黄芪、灵芝,由暨南大学生物医药基地提供,批号0929)或安慰剂(麦芽糊精),15g/次,连续服用30d。实验组测试当天6∶30服药1次。运动员在正式运动测试前进行5min的准备活动,休息5min后,开始以80%最大强度在划船测功仪上运动至力竭。采用ELITE流式细胞仪检测淋巴细胞数量和含量,CD4 T细胞、CD8 T细胞含量;采用ADVIACentaur化学发光免疫分析系统检测睾酮、皮质醇水平。结果:①淋巴细胞的变化:运动后24h,实验组受试者淋巴细胞含量、CD4 T细胞含量及CD4 与CD8 比值高于运动前及对照组,差异有显著性意义[分别为(41.36±4.80)%,(35.14±4.46)%,(34.08±3.45)%;(38.36±4.43)%,(31.71±4.98)%,(32.45±5.80)%;1.23±0.22,0.97±0.24,0.94±0.28,P<0.05]。②睾酮、皮质醇的变化:实验组运动后24h,血清睾酮、皮质醇水平低于运动前,差异无显著性意义[分别为(48.90±7.01),(55.58±9.25)ng/L;(1.48±0.20),(1.48±0.21)μg/L,P>0.05]。对照组运动后24h睾酮水平低于运动前,差异有显著性意义[分别为(49.01±8.16),(62.22±11.46)ng/L,P<0.05],皮质醇水平高于运动前,差异有显著性意义[分别为(1.88±0.20),(1.54±0.14)μg/L,P<0.05]。结论:扶正益气中药对大强度力竭运动后优秀赛艇运动员的CD4 与CD8 比值具有显著的升高作用,其作用机制之一可能为扶正益气中药很可能通过抑制运动后睾酮水平的降低及皮质醇水平的升高,达到提高CD4 T细胞含量,改善CD4 /CD8 比值,进而增强机体细胞免疫功能的作用。