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cruzi疗法副作用小[4]。每日两次剂量≥15mg\u002F kg体重,Posaconazole可延长小鼠的存活时间并减轻组织负担[5]。",{"key":253,"value":254},"细胞实验","寄生虫的epimastigote形式在肝输注胰蛋白胨培养基中培养，其中补充有10％新出生的小牛血清，在28℃下进行强烈（120rpm）搅拌。培养物以2×106 epimastigotes mL-1的细胞密度开始，并且以0.5-1.0×107 epimastigotes mL-1的细胞密度添加药物。通过使用电子粒子计数器以及通过血细胞计数器直接计数来测量细胞密度。细胞活力随后是台盼蓝排除，使用光学显微镜检查。如前所述，无鞭毛体在Vero细胞中培养，所述Vero细胞维持在补充有1％胎牛血清的最小必需培养基中，在37℃下在潮湿气氛（95％空气-5％CO 2）中。每个细胞用10个组织培养物衍生的trypomastigotes感染细胞2小时，然后用磷酸盐缓冲盐水（PBS）洗涤三次以除去非粘附寄生虫。添加含有和不含药物的新鲜培养基，将细胞培养96小时，培养基更换为48小时。使用光学显微镜直接测定感染细胞的百分比和每个细胞的寄生虫数量，并如前所述进行结果的统计分析。使用GraFit程序通过非线性回归计算IC 50值。对照和药物处理的细胞外epimastigotes中的细胞质游离Ca 2+浓度通过荧光测定法使用Fura-2测定，再次如前所述。通过使用时间扫描共聚焦显微镜，在用克氏锥虫无鞭毛体感染的个体Vero细胞上监测亚细胞Ca 2+水平和线粒体膜电位，如其他地方详细描述的。简而言之，用T. cruzi无鞭毛体严重感染（72小时）的Vero细胞接种到22×40mm玻璃盖玻片（0.15mm厚度）上，并与10μM细胞渗透性Rhod-2和10μg\u002F mL罗丹明-123同时孵育。在37℃下在培养基中50分钟，然后洗涤并与含有或不含有胺碘酮的林格氏溶液一起温育。在所使用的条件下，Rhod-2的荧光主要来自细胞内富含Ca2 +的区室，如线粒体，因为其对Ca2 +的低亲和力限制了其在Vero细胞或无鞭毛体的Ca2 +  - 细胞质中的荧光。罗丹明-123是一种线粒体特异性阳离子染料，严格根据其膜电位分布在线粒体内膜上。",{"key":256,"value":257},"动物实验","通过使用急性恰加斯病的鼠模型进行体内研究，其中雌性NMRI-IVIC小鼠（20-25g）用105或103个血流试验鞭毛体感染，并且在24小时后开始药物治疗。对于泊沙康唑（30剂）和\u002F或每隔一天50mg \u002F kg对于胺碘酮（15剂），以20mg \u002F kg \u002F d连续30天给予治疗。阴性对照（即未治疗的动物）仅接受载体，而阳性对照用抗-T治疗。克唑子化合物，硝呋莫司，50mg \u002F kg \u002F d，30天。每天进行生存，每周进行寄生虫血症，后者通过直接显微镜检查。在感染后观察动物60天，之后通过使用血培养，异种诊断和血液PCR测试的组合来评估寄生虫学治疗。",{"key":259,"value":260},"参考文献","[1].\nBenaim G, et al. Amiodarone has intrinsic anti-Trypanosoma cruzi activity and acts synergistically with posaconazole. J Med Chem. 2006 Feb 9;49(3):892-9.\n[2].\nSabatelli F, et al. In vitro activities of posaconazole, fluconazole, itraconazole, voriconazole, and amphotericin B against a large collection of clinically important molds and yeasts. Antimicrob Agents Chemother. 2006 Jun;50(6):2009-15.\n[3].\nSansone-Parsons A, et al. Effect of a nutritional supplement on posaconazole pharmacokinetics following oral administration to healthy volunteers. Antimicrob Agents Chemother. 2006 May;50(5):1881-3.\n[4].\nVeiga-Santos P, et al. Effects of amiodarone and posaconazole on the growth and ultrastructure of Trypanosoma cruzi. Int J Antimicrob Agents. 2012 Jul;40(1):61-71.\n[5].\nSun QN, et al. In vivo activity of posaconazole against Mucor spp. in an immunosuppressed-mouse model. 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