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（5×105），试图使它们适应MT-4细胞中的复制。在共培养的第7天（PHA-PBMC和MT-4细胞）收获无细胞上清液，并且病毒（称为HIV11MIXP0，其中P0代表第0代）在新鲜MT-4细胞中进一步繁殖用于选择实验。在第一次传代时，将MT-4细胞（5×105）暴露于混合病毒的培养上清液或每种感染性分子HIV-1克隆的500TCID50s，并在Tipranavir存在下以0.1至0.4μM的初始浓度培养。在每次传代的最后一天（大约第7天），收获1mL无细胞上清液，并在浓度增加的药物存在下转移到新鲜未感染的MT-4细胞培养物中，用于下一轮培养。 。在该轮培养中，使用三种药物浓度（与先前浓度相比增加1倍，2倍和3倍）。当通过大量Gag蛋白产生（大于200ng \u002F mL）证实培养物中HIV-1的复制时，三种浓度中的最高药物浓度用于继续选择（用于下一轮培养）。重复使用该方案直至药物浓度达到目标浓度。从感染细胞的裂解物中获得的前病毒DNA样品进行核苷酸测序[1]。",{"key":255,"value":256},"动物实验","小鼠[2]将所有小鼠（2-4个月大）维持在标准的12小时黑暗和12小时光照循环中，随意提供水和食物。对于代谢组学分析，通过球形管饲针施用替拉那韦（40mg \u002F kg），并将小鼠圈养在分开的代谢笼中18小时。收集尿液和粪便样品并储存在-20℃下用于进一步分析。对于组织分布和抑制研究，使用三组小鼠并用替拉那韦（100mg \u002F kg），RTV（40mg \u002F kg）和替拉那韦\u002F r（100mg \u002F kg替拉那韦和40mg \u002F kg RTV）口服治疗。 ）， 分别。处理后30分钟收集包括肝，脑，肺，肾，脾和眼的组织，并在-20℃下储存用于进一步分析。",{"key":258,"value":259},"参考文献","[1].\nAoki M, et al. 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