32只健康本地犬饲养1周后按体质量灌胃番泻叶水煎剂6 m L/kg人工复制脾虚泄泻证病理模型,造模成功后随机分为4组:黄芪多糖治疗组后海穴注射黄芪多糖注射液0.25 m L/kg,2次/d;硫酸庆大霉素治疗组后海穴注射硫酸庆大霉素注射液0.2 m L/kg,2次/d;后海穴针刺组后海穴注射生理盐水0.25 m L/kg,2次/d;病理对照组不作任何处理;同时设立4头健康犬作为空白对照,造模期间灌胃生理盐水。用药后治疗期间测定试验犬只的体质量、三大临床指标、部分血液指标和脾脏指数,并统计分析。结果表明,黄芪多糖治疗组血红蛋白、红细胞水平在较短时间内恢复正常,脾脏指数显著(P<0.05)高于硫酸庆大霉素治疗组;硫酸庆大霉素治疗组白细胞降低速度极显著(P<0.01)快于黄芪多糖治疗组。这表明后海穴注射黄芪多糖注射液可以用于治疗犬脾虚泄泻证。
[Objective] This study aimed to optimize the PCR amplification conditions for random ssDNA pool in SELEX technology. [Method] L16（45） orthogonal experimental design was adopted for optimization of five important factors affecting PCR reaction system for random single-stranded DNA pool including Mg2＋ concentration, dNTP concentration, amount of Taq DNA polymerase, primer concentration and amount of random single-stranded DNA pool at four levels. Meanwhile, the annealing temperature and number of PCR reaction cycles were optimized to establish the optimal reaction system and PCR procedure. [Result] The optimal combination of PCR reaction system for random ssDNA pool was obtained, with a total system volume of 20 μl containing 2.0 μl of 10 × Buffer, 0.5 ng of random ssDNA pool, 2.5 mmol/L Mg2＋, 0.25 mmol/L dNTP Mixture, 0.6 μmol/L upstream and downstream primers and 1.5 U of Taq DNA polymerase; the optimal annealing temperature was 68 ℃ and the optimal number of cycles was 12. Under the above conditions, clear and stable bands with high specificity for random ssDNA pool were amplified. [Conclusion] This study laid the foundation for selection of parameters with higher specificity in SELEX technology.
