蒿甲醚对小鼠体内日本血吸虫的己糖激酶、磷酸葡萄糖异构酶和磷酸果糖激酶的影响(英文)

目的：研究蒿甲醚（Ａｒｔ）对血吸虫己糖激酶（ＨＫ）、磷酸葡萄糖异构酶（ＧＰＩ）和磷酸果糖激酶（ＰＦＫ）的影响。方法：感染血吸虫小鼠１次ｉｇＡｒｔ１００或３００ｍｇｋｇ－１，并分别于２４ｈ和４８ｈ剖杀取虫。按生成ＮＡＤＰＨ和耗用ＮＡＤＨ的方法测定上述３种酶活力。结果：感染鼠ｉｇ．Ａｒｔ３００ｍｇｋｇ－１后２４ｈ，♀、虫体ＨＫ活力抑制率分别为３３．３％和１３．７％，ＧＰＩ活力则分别抑制１４．７％和１７．５％，４８ｈ后，♀、虫体的ＧＰＩ活力抑制率分别为４６．２％和３２．９％。但Ａｒｔ剂量为１００或３００ｍｇｋｇ－１时，给药后２４ｈ和４８ｈ的♀虫ＰＦＫ抑制率分别为６４．９％和７１％，均明显高于虫的１６．３％和５４．２％。结论：ＰＦＫ可能是Ａｒｔ作用于血吸虫糖酶解途径的靶酶之一。     

蒿甲醚对小鼠体内日本血吸虫的己糖激酶、磷酸葡萄糖异构酶和磷酸果糖激酶的影响

目的：研究蒿甲醚（Ａｒｔ）对血吸虫己糖激酶（ＨＫ）、磷酸葡萄糖异构酶（ＧＰＩ）和磷酸果糖激酶（ＰＦＫ）的影响。方法：感染血吸虫小鼠１次ｉｇＡｒｔ１００或３００ｍｇｋｇ－１,并分别于２４ｈ和４８ｈ剖杀取虫。按生成ＮＡＤＰＨ和耗用ＮＡＤＨ的方法测定上述３种酶活力。结果：感染鼠ｉｇ．Ａｒｔ３００ｍｇｋｇ－１后２４ｈ,♀、虫体ＨＫ活力抑制率分别为３３．３％和１３．７％,ＧＰＩ活力则分别抑制１４．７％和１７．５％,４８ｈ后,♀、虫体的ＧＰＩ活力抑制率分别为４６．２％和３２．９％。但Ａｒｔ剂量为１００或３００ｍｇｋｇ－１时,给药后２４ｈ和４８ｈ的♀虫ＰＦＫ抑制率分别为６４．９％和７１％,均明显高于虫的１６．３％和５４．２％。结论：ＰＦＫ可能是Ａｒｔ作用于血吸虫糖酶解途径的靶酶之一。     

蒿甲醚对日本血吸虫糖原,蛋白质,硷性磷酸酶和…

感染日本血吸虫尾蚴３２－３５ｄ的小鼠１次ｉｇ蒿甲醚（Ａｒｔ）３００ｍｇ．ｋｇ＾－＾１后２４ｈ，其体内♀，♂血吸虫的糖原含量较对照组减少约５０％，７２ｈ后则分别减少６４．１％和７７．９％。给药后７２ｈ，♀，♂虫的蛋白质含较对照组分别减少６８．．１％和４９．３％。给药２４ｈ，♀，♂虫的硷性磷酸酶（ＡＫＰ）活力的抑制率约为２５％，７２ｈ后，♀虫的ＡＫＰ进一步被抑制，抑制率达６１．６％。酸性磷酸酶（ＡＣＰ     

用流式细胞术分析双氢青蒿素对杜氏利什曼原虫DNA的作用

应用流式细胞术（ Ｆ Ｃ Ｍ ）检测分析了双氢青蒿素对杜氏利什曼原虫前鞭毛体 Ｄ Ｎ Ａ 的作用。４ 个不同浓度（１７７×１０－ ４,３５３×１０－ ４,７１×１０－ ４,１４１×１０４ｍ ｏｌ／ Ｌ）的药物对虫体 Ｄ Ｎ Ａ 均有抑制作用,用药组虫体 Ｄ Ｎ Ａ 含量的荧光分布随着药物浓度的升高而减弱,抑制率分别为 ３２３％ ,３２６％ ,８６４％ ,８９２％ ,与对照组相比有显著性差异（ Ｐ＜ ００５）。后两个用药组与前两个用药组抑制率有显著性差异（ Ｐ＜ ０００５）。本研究结果与光镜观察、３ Ｈ Ｔｄ Ｒ 掺入试验结果一致     

蒿甲醚对日本血吸虫糖原、蛋白质、硷性磷酸酶和酸性磷酸酶的影响

感染日本血吸虫尾蚴３２－３５ｄ的小鼠１次ｉｇ蒿甲醚（Ａｒｔ）３００ｍｇ·ｋｇ－１后２４ｈ，其体内♀、血吸虫的糖原含量较对照组减少约５０％，７２ｈ后则分别减少６４．１％和７７．９％。给药后７２ｈ，♀、虫的蛋白质含量较对照组分别减少６８．１％和４９．３％。给药２４ｈ，♀、虫的硷性磷酸酶（ＡＫＰ）活力的抑制率约为２５％，７２ｈ后，♀虫的ＡＫＰ进一步被抑制，抑制率达６１．６％。酸性磷酸酶（ＡＣＰ）的活力受抑制亦以♀虫较为明显，给药７２ｈ♀、虫抑制率分别为７５．７％和４７．６％。     

日本血吸虫3-磷酸甘油醛脱氢酶的抗原性和保护性免疫力的研究

３－磷酸甘油醛脱氢酶（ＧＡＰＤＨ），是血吸虫糖酵解中的重要酶，参与虫体内多种代谢功能，在抗血吸虫疫苗的研制中亦受到重视曼氏血吸虫ＧＡＰＤＨ３７ｋＤａ抗原性与保护性免疫力已被证实［１］，日本血吸虫菲律宾株ＧＡＰＤＨ克隆及具有生物活性重组体的特性与免疫原...     

日本血吸虫副肌球蛋白全基因克隆、测序及体内表达

目的： 将日本血吸虫大陆株副肌球蛋白（ Ｓｊｃ９７） 编码区全基因克隆及测序, 并研究 Ｓｊｃ９７ 全基因核酸疫苗在小鼠体内的表达。方法： 抽提日本血吸虫成虫总 Ｒ Ｎ Ａ, 逆转录 聚合酶链反应 （ Ｒ Ｔ Ｐ Ｃ Ｒ） 扩增编码 Ｓｊｃ９７的全基因ｃ Ｄ Ｎ Ａ, 双脱氧链末端终止法测 Ｄ Ｎ Ａ 序列。构建含 Ｓｊｃ９７ 全基因的质粒表达载体 （ｐ Ｃ Ｍ Ｖ Ｓｊｃ９７） , 并用免疫荧光染色法研究ｐ Ｃ Ｍ Ｖ Ｓｊｃ９７ 在小鼠体内的表达特性。结果与结论： 用 Ｒ Ｔ Ｐ Ｃ Ｒ 扩增获得了 Ｓｊｃ９７ 的编码区全基因ｃ Ｄ Ｎ Ａ, 并测定了该基因编码区的全序列。与日本血吸虫菲律宾株、日本株及曼氏血吸虫副肌球蛋白编码区全基因比较, 核苷酸序列同源性分别为： ９９４ ％ 、９９２ ％ 和９１０ ％ ; 推导的氨基酸序列同源性分别为：９９７ ％ 、９９８ ％ 和９６０ ％ 。ｐ Ｃ Ｍ Ｖ Ｓｊｃ９７ 核酸疫苗能在注射的小鼠局部肌肉组织表达 Ｓｊｃ９７ 。     

不同化疗方案治疗老年人急性髓系白血病疗效观察

４５例老年人急性髓系白血病经不同化疗方案治疗，ＨＡ方案剂量个体化组治疗１９例，完全缓解（ＣＲ）率５２６％；ＤＡ方案组治疗８例，ＣＲ率５０％；ＨＯＡＰ方案组治疗９例，ＣＲ率４４４％；小剂量阿糖胞苷（ＬＤＡｒａＣ）组治疗９例，ＣＲ率１１１％。ＨＡ方案剂量个体化组ＣＲ率显著高于ＬＤＡｒａＣ组（Ｐ＜００５）。骨髓抑制的发生率和治疗相关病死率分别为３６８％和５６％（ＨＡ）、７５％和３７５％（ＤＡ）、４４４％和２２２％（ＨＯＡＰ）、３３３％和１１１％（ＬＤＡｒａＣ），ＤＡ方案组治疗相关病死率显著高于ＨＡ方案剂量个体化组（Ｐ＜００２５）。联合化疗治疗３６例，ＣＲ率为５０％，高于单用ＬＤＡｒａＣ组（Ｐ＜００５），骨髓抑制发生率为４７２％（１７／３６），治疗相关病死率为１６７％（６／３６），同ＬＤＡｒａＣ组比较无显著性（Ｐ＞００５）。     

武汉地区Ⅱ型糖尿病患者冠心病与载脂蛋白E基因型的关系

应用聚合酶链反应（ＰＣＲ）技术，对随机选择的１２５例ＮＩＤＤＭ患者和５０例非ＤＭ患者进行ＡｐｏＥ基因型检测，以研究ＮＩＤＤＭ患者ＣＨＤ与ＡｐｏＥ基因型间的关系。结果表明，ＮＩＤＤＭ患者心肌梗塞和缺血性心电图改变在Ａｐｏε４／４和ε４／３型组中发生率分别为２１％和４１％，但不同基因型组间差异无显著性。心绞痛在Ａｐｏε４／４和ε４／３型组中为５２％，显著高于ε３／３型组（３１％，Ｐ＜０．０５）。Ａｐｏε４／４和ε４／３型ＮＩＤＤＭ患者，任何证据的ＣＨＤ发生率为７２％，显著高于ε３／３型组（３７％）及ε２／２、ε３／２型组（３３％，Ｐ＜０．０１）。ＮＩＤＤＭ患者中ＣＨＤ组Ａｐｏε３／３和ε４／３基因型频率分别为５２％和３４％，分别低于（ε３／３）、高于（ε４／３）非ＣＨＤ组（７１％，Ｐ＜０．０５；１０％，Ｐ＜０．０１）；ＣＨＤ组ε４等位基因频率为２１％，明显高于非ＣＨＤ组（７％，Ｐ＜０．０１）。提示Ａｐｏε４／４和ε４／３为ＮＩＤＤＭ患者ＣＨＤ的重要危险指标。     

蒿甲醚早期治疗鼠血吸虫病的疗效

蒿甲醚系青蒿素的衍生物，不仅有抗疟作用，还用抗血吸虫（特别是ｄ７童虫）作用。本文应用Ａｒｔ的早期治疗方案，观察其控制血吸虫感染的可能性。结果表明小鼠于感染日本血吸虫尾蚴当天（ｄ０）ｉｇＡｒｔ３００ｍｇ．ｋｇ＾－１，然后每隔２－３ｗｋｉｇ１次相同剂量的的Ａｒｔ时，减早率和减雌虫率均低，若于感染后ｄ７开始ｉｇ首剂Ａｒｔ然后每ｗｋｉｇ１次，共给４次时，可使宿主体内的雌虫减少近９０％或９０％以上，且部分鼠     

青蒿琥酯抗日本血吸虫童虫和成虫作用的研究

目的 观察青蒿琥酯对不同阶段日本血吸虫的损害作用,揭示青蒿琥酯的杀虫机制.方法小鼠感染尾蚴18 d时给予青蒿琥酯300 mg/kg,24 h后灌注法收集童虫,并用紫外吸收法和Bradford标准曲线法测定青蒿琥酯对日本血吸虫18 d童虫DNA和蛋白质含量的影响;分别将10条虫体在含有3H胸腺嘧啶核苷的培养基中培养24 h后,用滤膜法和匀浆法测定青蒿琥酯对标记核苷掺入童虫DNA的影响;感染血吸虫尾蚴21 d的小鼠一次性灌服青蒿琥酯300 mg/kg,给药后21 d,灌注法收集成虫,观察青蒿琥酯对日本血吸虫成虫生殖器官大小及生殖细胞发育的影响;小鼠感染血吸虫33 d时给予同样的青蒿琥酯进行治疗,24 h和48 h后处死小鼠,将肝脏制作组织切片,在光镜下观察同等剂量青蒿琥酯引起的成虫肝移和形态学变化的影响.结果 青蒿琥酯体内作用18 d的虫体,24 h后,童虫的DNA和蛋白质含量均较对照组明显减少,减少率分别为23.0%和33.6%(P＜0.01);在含有3H胸腺嘧啶核苷的培养基中培养24 h后,童虫摄入标记的核苷及核苷掺入童虫DNA的量较对照组也明显减少,减少率分别为61.1%和40.8%(P＜0.01);21 d给药组小鼠体内雌虫卵巢的成熟、发育受到了抑制;药物作用于虫体33 d后能明显引起虫体形态的变化,特别是虫体体表.结论 青蒿琥酯能减少日本血吸虫童虫蛋白质和DNA的含量,能明显抑制虫体对标记核苷的摄入以及标记核苷掺入虫体DNA的量,能引起虫体体表明显的组织形态学变化,并且能抑制雌虫卵巢的发育,降低或抑制雌虫的产卵,能有效减轻血吸虫病的病情和控制传播.     

蒿甲醚对日本血吸虫磷酸葡萄糖变位酶、醛缩酶、磷酸甘油酸变位酶和烯醇化酶的影响

[目的 ]观察蒿甲醚 (Art)对小鼠体内日本血吸虫磷酸葡萄糖变位酶 (GPM )、醛缩酶 (ALD)、磷酸甘油酸变位酶 (PGM)和烯醇化酶 (ENO)的影响。 [方法 ]小鼠感染血吸虫尾蚴 4～ 5wk后 ,1次灌服Art10 0mg/kg或 30 0mg/kg ,并于 2 4～ 48h后剖杀 ,收集日本血吸虫雌虫和雄虫 ,按NADPH的生成量或NADH的耗用量测定虫体的上述 4种酶活力。 [结果 ]经Art 10 0mg/kg作用 2 4h后 ,雌虫的GPM、ALD、PGM和ENO活力分别较对照组下降 15 %、 19%、 5 0 %和 46 % ,其间差别均具有显著意义 ,而雄虫仅PGM和ENO活力分别下降 2 2 %和 32 % ;48h后 ,雄虫的GPM和ALD活力亦分别下降 2 1%和 18% ,雌虫的GPM、ALD、PGM和ENO及雄虫的PGM和ENO活力则进一步下降。经Art 30 0mg/kg作用后 2 4～ 48h ,雌虫和雄虫的上述 4种酶活力均明显下降 ,且呈一定的时间效应关系。 [结论 ]Art对日本血吸虫尤其是雌虫的上述 4种酶有抑制作用。     

蒿甲醚诱导日本血吸虫雌虫总抗氧化能力下降(英)

目的 观察蒿甲醚对日本血吸虫成虫总抗氧化能力的影响。 方法 体外将血吸虫在含蒿甲醚和氯化血红素的培养液内培养24 h后,或体内感染小鼠经蒿甲醚300mg/kg治疗6～24 h后,测定虫体的总抗氧化能力。 结果体外50μmol/L的蒿甲醚与氯化血红素伍用引起雌虫总抗氧化能力明显下降。体内蒿甲醚作用血吸虫6 h,即见雌虫的总抗氧化能力明显下降。体内、体外试验中,蒿甲醚对雄虫的总抗氧化能力均无影响。 结论 蒿甲醚诱导雌虫总抗氧化能力下降。     

Oltipraz-induced reduction in schistosomal glucose utilization rates

The rate of phosphorylation of 2-deoxyglucose (2DG) was determined by sequential pulsing of schistosomes (male and female Schistosoma mansoni) with [3H- and 14C]-2-deoxy-D-glucose. The relative phosphorylation rate of 2-[3H]-2DG to 1-[14C]-D-glucose (i.e. the phosphorylation coefficient) was also measured in male and female schistosomes. Even though 2DG is taken up more rapidly than glucose, it is phosphorylated at a much slower rate in S. mansoni. Mated schistosomes phosphorylate 2DG and glucose at a greater rate than do unmated worms. In contrast, the phosphorylation coefficient is greater in separated than mated schistosomes. In schistosomes exposed to oltipraz for short time periods (6 min, at a concentration of 10 micrograms/ml) glucose utilization rates were significantly reduced in (both mated and separated) female S. mansoni and by a similar magnitude (not significant) in males.     

Dipyridamole inhibits reversion by thymidine of methotrexate effect and increases drug uptake in Sarcoma 180 cells.

The combined effect of methotrexate (MTX) with dipyridamole, an inhibitor of nucleoside transport, was studied in ascitic Sarcoma 180 cells. It was determined that 10 microM MTX inhibits by greater than 90% deoxy[3H]uridine incorporation into DNA and that this MTX concentration inhibits DNA synthesis as revealed by deoxy[3H]cytidine but not [3H]thymidine incorporation into DNA. Exogenous thymidine (greater than or equal to 1 microM) in the cell culture medium enhances DNA synthesis in nontreated cells and fully restores it in MTX-treated cells, whereas hypoxanthine has no appreciable effect on DNA synthesis. Dipyridamole inhibits deoxy[3H]cytidine and [3H]thymidine uptake by these cells (IC50 = 0.2 and 3 microM, respectively) and blocks the increase in TTP pool produced by 1 microM thymidine in MTX-treated cells (23.1 +/- 4.7 pmol per 1 X 10(6) cells vs. 80.4 +/- 18.9 pmol per 1 X 10(6) cells). Dipyridamole at 10 microM enhances [3H]MTX accumulation by Sarcoma 180 cells and diminishes the efflux of the drug in previously loaded cells. It is suggested that the combination of inhibitors of the de novo pathway for pyrimidine biosynthesis, such as MTX, with inhibitors of the salvage pathway, such as dipyridamole, may increase the cytotoxic activity of MTX alone.     

Insulin-like growth factor I has independent effects on bone matrix formation and cell replication

The effects of insulin-like growth factor-I (IGF-I) and insulin on bone matrix synthesis and bone cell replication were studied in cultured 21-day-old fetal rat calvariae. Histomorphometry techniques were developed to measure the incorporation of [2,3-3H]proline and [methyl-3H]thymidine into bone matrix and bone cell nuclei, respectively, using autoradiographs of sagittal sections of calvariae cultured with IGF-I, insulin, or vehicle for up to 96 h. To confirm an effect on bone formation, IGF-I was also studied for its effects on [3H]proline incorporation into collagenase-digestible protein (CDP) and noncollagen protein and on [3H]thymidine incorporation into acid-precipitable material (DNA). IGF-I at 10(-9)-10(-7) M significantly increased the rate of bone matrix apposition and CDP after 24 h by 45-50% and increased cell labeling by 8-fold in the osteoprogenitor cell zone, by 4-fold in the osteoblast cell zone, and by 2-fold in the periosteal fibroblast zone. Insulin at 10(-9)-10(-6) M also increased matrix apposition rate and CDP by 40-50%, but increased cell labeling by 2-fold only at a concentration of 10(-7) M or higher and then only in the osteoprogenitor cell zone. When hydroxyurea was added to IGF-I-treated bones, the effects of IGF-I on DNA synthesis were abolished, but the increase in bone matrix apposition induced by IGF-I was only partly diminished. In conclusion, IGF-I stimulates matrix synthesis in calvariae, an effect that is partly, although not completely, dependent on its stimulatory effect on DNA synthesis.     

Actions of insulin and hydrocortisone on macromolecular synthesis in primary epithelial cell cultures from mouse mammary glands.

Monolayer cultures of mammary gland epithelial cells were prepared from the abdominal glands of midpregnancy mice. After collagenase digestion of mammary tissue and separation by differential centrifugation, the isolated epithelial cells were cultured in Eagle's Minimal Essential Medium supplemented with 10% fetal bovine serum and insulin (6 micrograms/ml). Six days later, when the cultures were in log growth and nearly confluent, the effects of insulin and/or hydrocortisone on the rates of RNA, DNA, and protein synthesis were determined in a serum-free medium. At physiological concentrations, insulin enhanced the rates of uptake and incorporation of [3H]uridine into RNA, of [3H]thymidine into DNA, and of [3H]leucine into protein. Hydrocortisone was shown to be biphasic with regard to concentration in attenuating or augmenting insulin's effects on macromolecular synthesis.     

Organic iodine inhibits deoxyribonucleic acid synthesis and growth in FRTL-5 thyroid cells

FRTL-5 thyroid epithelial cells in culture were used to study the possible inhibitory effects of iodine on thyroid growth. NaI exerted a dose-dependent, thyroid epithelial cell-specific inhibitory effect on [methyl-3H]thymidine incorporation into DNA, reduced the DNA content in the cell layer, and limited the increase in cell number mediated by TSH. The inhibitory effects of sodium iodide applied to growth stimulated by TSH-, cAMP-, and non-cAMP-dependent mechanisms and were prevented by 1-methylimidazole-2-thiol (methimazole) and 2-ethylthioisonicotinamide (ethionamide). The latter findings indicate that the inhibitory effects of NaI are mediated by some iodine-containing organic compound. The inhibitory effects of organic iodine on growth subsided 24-48 h after removal of excess NaI from the culture medium. In contrast, NaI had no effect on normal rat kidney fibroblast or thyroid fibroblast [methyl-3H]thymidine incorporation stimulated by epidermal growth factor or serum. These data demonstrate a specific inhibitory effect of organic iodine on thyroid epithelial cell growth.     

Aging of FRTL-5 rat thyroid cells causes sensitivity to cytotoxicity induced by tumor necrosis factor-alpha.

While investigating the modulation of the growth and function of the FRTL-5 rat thyroid cell line by recombinant human tumor necrosis factor-alpha (TNF alpha), we noticed that pronounced changes in several response parameters occurred with increasing passage number. For young cells (passage less than 20), TNF alpha by itself slightly increased [3H]thymidine incorporation and DNA content, and had a minimal effect on basal 125I uptake. When combined with TSH, TNF alpha had no influence on TSH-stimulated [3H]thymidine incorporation, but significantly inhibited TSH-stimulated 125I uptake. Compared with young cells, aged cells (passage greater than 40), in contrast, developed a high sensitivity to TNF alpha. TNF alpha markedly stimulated [3H]thymidine incorporation into DNA, inhibited TSH-stimulated 125I uptake per micrograms DNA, but dramatically decreased the total DNA content and cell number. TSH augmented the TNF alpha effect in aged cells, resulting in a further reduction of DNA content. Aphidicolin, a specific inhibitor of DNA polymerase-alpha which is associated with DNA replication, dramatically inhibited TNF alpha-induced [3H]thymidine incorporation in both young and aged cells; this suggested that the effect of TNF alpha on FRTL-5 cell growth is related to DNA replication, rather than DNA repair. 51Cr release from FRTL-5 cells, a measure of cytotoxicity, increased 2-fold over baseline in aged cells at a dose of 400 ng/ml TNF alpha and decreased to 70% of baseline in young cells at this same dose. The protein kinase-A (PKA) and protein kinase-C (PKC) signal transduction mechanisms of TNF alpha in aged cells (passage greater than 40) were also studied. TNF alpha increased cAMP and also increased relative PKA and PKC activity in 1-40 min. Phorbol myristate acetate (PMA), an activator of PKC, increased [3H]thymidine incorporation and DNA content. PMA did not affect the TNF alpha-induced increase in [3H]thymidine incorporation or its reduction of DNA content. When the cells were pretreated with a high concentration of PMA (1 microM/24 h) to down-regulate PKC, the TNF alpha dose-dependent increase in [3H]thymidine incorporation and decrease in DNA content were only slightly inhibited, suggesting that the main effects of TNF alpha are independent of PKC. We conclude that the sensitivity of FRTL-5 cells to the cytotoxic effect of TNF alpha increases with aging.