Efficient drug screening of advanced larval stages of <i>S</i>. <i>mansoni</i> generated in human serum.

NTS were cultured in HM supplemented with 200 U/ml Penicillin and 200 μg/ml Streptomycin and 20% HSe for 1 week to generate LuS (B, D, F and H) and 6 weeks to obtain LiS (A, C, E, G) schistosomula. (A, B) Praziquantel (PZQ), (C, D) Oxamniquine (OXM), (E, F) Mefloquine (MFQ) and (G, H) Artemether (ART) were dissolved in DMSO and added at indicated concentrations for the entire duration of the experiment. 1% DMSO in culture medium served as control. Viability was scored before treatment (0 h) and 3 hours, 1, 2, 3 and 7 days a.t. Each data point is shown as a mean ± SD of three independent experiments with at least three biological replicates each. ××× p ≤ 0.001, ×× p ≤ 0.01, × p ≤ 0.05 comparing control with 1 μg/ml drug; ┴┴┴ p ≤ 0.001 ┴┴ p ≤ 0.01 ┴ p ≤ 0.05 control with 10 μg/ml drug; ***p ≤ 0.001, **p ≤ 0.01 *p ≤ 0.05 control with 100 μg/ml drug. LuS, lung stage; LiS, liver stage; s.p., score points; a.t., after treatment. (PZFX)将NTS置于添加200 U/ml青霉素（Penicillin）、200 μg/ml链霉素（Streptomycin）及20% HSe的HM培养基中培养，培养1周可制备肺期血吸虫童虫（LuS，对应B、D、F、H组），培养6周则可获得肝期血吸虫童虫（LiS，对应A、C、E、G组）。A、B组为吡喹酮（Praziquantel, PZQ）处理组，C、D组为奥沙尼喹（Oxamniquine, OXM）处理组，E、F组为甲氟喹（Mefloquine, MFQ）处理组，G、H组为蒿甲醚（Artemether, ART）处理组；将各药物溶于二甲基亚砜（DMSO）中，于实验全程按指定浓度添加至培养基，以含1% DMSO的培养基作为空白对照。分别于给药前（0 h）及给药后（a.t.）3 h、1 d、2 d、3 d和7 d对虫体活力进行评分。所有数据点均以三次独立实验的平均值±标准差（SD）表示，每次独立实验至少设置三个生物学重复。以对照组为参照，与1 μg/ml药物处理组相比，×××对应p ≤ 0.001、××对应p ≤ 0.01、×对应p ≤ 0.05；与10 μg/ml药物处理组相比，┴┴┴对应p ≤ 0.001、┴┴对应p ≤ 0.01、┴对应p ≤ 0.05；与100 μg/ml药物处理组相比，***对应p ≤ 0.001、**对应p ≤ 0.01、*对应p ≤ 0.05。其中LuS即肺期（lung stage），LiS即肝期（liver stage），s.p.为评分点数（score points），a.t.为给药后（after treatment）。(PZFX)