目的观察伯氏疟原虫静息巯基氧化酶(PbQSOX)的表达阶段及表达特点。方法分别采用SignalP-5.0 Server与TMHMM Server 2.0在线软件对PbQSOX蛋白的信号肽与跨膜区进行预测分析。利用同源重组技术,将PbQSOX-HA标签型打靶质粒经限制性内切酶...目的观察伯氏疟原虫静息巯基氧化酶(PbQSOX)的表达阶段及表达特点。方法分别采用SignalP-5.0 Server与TMHMM Server 2.0在线软件对PbQSOX蛋白的信号肽与跨膜区进行预测分析。利用同源重组技术,将PbQSOX-HA标签型打靶质粒经限制性内切酶NotⅠ酶切使其线性化,将线性化的质粒电转染至伯氏疟原虫株内并感染小鼠。经乙胺嘧啶筛选耐药型疟原虫血症小鼠,取小鼠外周血进行PCR鉴定,获取PbQSOX-HA标签型疟原虫。培养纯化裂殖体、配子体与动合子阶段的PbQSOX-HA标签型疟原虫,采用蛋白质印迹法与间接免疫荧光实验检测PbQSOX蛋白在伯氏疟原虫中的表达阶段与表达特点。结果生物信息学分析发现PbQSOX蛋白存在信号肽,不存在跨膜区,蛋白均在细胞膜外表达。经PCR鉴定,PbQSOX-HA标签型打靶质粒与伯氏疟原虫同源重组正确,并成功获得了PbQSOX-HA标签型疟原虫及纯化的PbQSOX-HA标签型裂殖体、配子体与动合子。蛋白质印迹法检测显示PbQSOX蛋白主要在配子体与动合子阶段表达,间接免疫荧光实验结果显示PbQSOX表达于配子体、雄配子、合子、retort与动合子表面。结论PbQSOX是伯氏疟原虫在有性生殖阶段表达的蛋白,主要表达于配子体、雄配子、合子、retort与动合子表面。展开更多
目的:研究疟原虫抗青蒿酯钠的产生机理。方法:分别采用感染抗青篙酯钠伯氏疟原虫株小鼠血清(SM SA-R)、感染青蒿酯钠敏感株伯氏疟原虫小鼠血清(SM SA-S)和无原虫感染的正常小鼠血清(SM)与青蒿酯钠(SA)水溶液的混合物治疗感染正常伯氏疟...目的:研究疟原虫抗青蒿酯钠的产生机理。方法:分别采用感染抗青篙酯钠伯氏疟原虫株小鼠血清(SM SA-R)、感染青蒿酯钠敏感株伯氏疟原虫小鼠血清(SM SA-S)和无原虫感染的正常小鼠血清(SM)与青蒿酯钠(SA)水溶液的混合物治疗感染正常伯氏疟原虫小鼠,于给药后3天(d3)、4天(d4)和5天(d5)涂片检查其感染率。结果:SM SA-R 加 SA 组、SMSA-S 加 SA 组和 SM 加 SA 组的原虫感染率分别为:d3:(16.8±1.7)%、(10.3±2.5)%、(9.0±2.0)%,d4:(28.6±5.0)%、(20.9±5.2)%、(17.6±4.4)%,d5:(39.2±8.5)%、(27.0±7.1)%、(23.1±5.4)%。SM SA-R 加 SA 组与其他两组原虫感染率比较,差异显著(P<0.01)。结论:SM SA-R 对 SA 抗疟药药效具有拮抗作用。提示 SA-R 疟原虫可能产生了某种“特异性抗药物质”。展开更多
Objective:To evaluate the in rivo antiplasmodial activities of the extract and fractions(n-hexane. chloroform,ethylacetate.butanol.aqueous) of the whole plant in Plasmodium berghei berghei infected mice.Methods:Oral a...Objective:To evaluate the in rivo antiplasmodial activities of the extract and fractions(n-hexane. chloroform,ethylacetate.butanol.aqueous) of the whole plant in Plasmodium berghei berghei infected mice.Methods:Oral administrations of the extract(200.400,and 600 my/kg) of Eleacine indica and fractions(400 mg/kg) were screened in the 4-day.repository and curative tests. Chloroquine(5 mg/kg).pyrimethamine(1.2 mg/kgi and artesunate(5 mg/kgi were used as controls. Results:The extract showed significant[P【 0.05-0.001) dose-dependent,antiplasmodial activity in the 4-day.repository and curative tests and increased the survival times of the infected mice. All the fractions exhibited significant antiplasmodial activity with the highest being ethylacetate fraction.Conclusions:Eleucine indica extract and fractions possess antimalarial activity which confirms the ethnobotanical use of this plant as a malarial remedy and opens a new highway to further investigate its potentials in the on-going fight against malaria.展开更多
文摘目的观察伯氏疟原虫静息巯基氧化酶(PbQSOX)的表达阶段及表达特点。方法分别采用SignalP-5.0 Server与TMHMM Server 2.0在线软件对PbQSOX蛋白的信号肽与跨膜区进行预测分析。利用同源重组技术,将PbQSOX-HA标签型打靶质粒经限制性内切酶NotⅠ酶切使其线性化,将线性化的质粒电转染至伯氏疟原虫株内并感染小鼠。经乙胺嘧啶筛选耐药型疟原虫血症小鼠,取小鼠外周血进行PCR鉴定,获取PbQSOX-HA标签型疟原虫。培养纯化裂殖体、配子体与动合子阶段的PbQSOX-HA标签型疟原虫,采用蛋白质印迹法与间接免疫荧光实验检测PbQSOX蛋白在伯氏疟原虫中的表达阶段与表达特点。结果生物信息学分析发现PbQSOX蛋白存在信号肽,不存在跨膜区,蛋白均在细胞膜外表达。经PCR鉴定,PbQSOX-HA标签型打靶质粒与伯氏疟原虫同源重组正确,并成功获得了PbQSOX-HA标签型疟原虫及纯化的PbQSOX-HA标签型裂殖体、配子体与动合子。蛋白质印迹法检测显示PbQSOX蛋白主要在配子体与动合子阶段表达,间接免疫荧光实验结果显示PbQSOX表达于配子体、雄配子、合子、retort与动合子表面。结论PbQSOX是伯氏疟原虫在有性生殖阶段表达的蛋白,主要表达于配子体、雄配子、合子、retort与动合子表面。
文摘目的:研究疟原虫抗青蒿酯钠的产生机理。方法:分别采用感染抗青篙酯钠伯氏疟原虫株小鼠血清(SM SA-R)、感染青蒿酯钠敏感株伯氏疟原虫小鼠血清(SM SA-S)和无原虫感染的正常小鼠血清(SM)与青蒿酯钠(SA)水溶液的混合物治疗感染正常伯氏疟原虫小鼠,于给药后3天(d3)、4天(d4)和5天(d5)涂片检查其感染率。结果:SM SA-R 加 SA 组、SMSA-S 加 SA 组和 SM 加 SA 组的原虫感染率分别为:d3:(16.8±1.7)%、(10.3±2.5)%、(9.0±2.0)%,d4:(28.6±5.0)%、(20.9±5.2)%、(17.6±4.4)%,d5:(39.2±8.5)%、(27.0±7.1)%、(23.1±5.4)%。SM SA-R 加 SA 组与其他两组原虫感染率比较,差异显著(P<0.01)。结论:SM SA-R 对 SA 抗疟药药效具有拮抗作用。提示 SA-R 疟原虫可能产生了某种“特异性抗药物质”。
基金supported by research grant from Education Trust Fund of University of Uyo,Uyo
文摘Objective:To evaluate the in rivo antiplasmodial activities of the extract and fractions(n-hexane. chloroform,ethylacetate.butanol.aqueous) of the whole plant in Plasmodium berghei berghei infected mice.Methods:Oral administrations of the extract(200.400,and 600 my/kg) of Eleacine indica and fractions(400 mg/kg) were screened in the 4-day.repository and curative tests. Chloroquine(5 mg/kg).pyrimethamine(1.2 mg/kgi and artesunate(5 mg/kgi were used as controls. Results:The extract showed significant[P【 0.05-0.001) dose-dependent,antiplasmodial activity in the 4-day.repository and curative tests and increased the survival times of the infected mice. All the fractions exhibited significant antiplasmodial activity with the highest being ethylacetate fraction.Conclusions:Eleucine indica extract and fractions possess antimalarial activity which confirms the ethnobotanical use of this plant as a malarial remedy and opens a new highway to further investigate its potentials in the on-going fight against malaria.
