吡喹酮和硝硫氰醚对日本血吸虫体内5-HT含量的影响

本文研究了吡喹酮和硝硫氰醚与日本血吸虫拟似神经递质 5-羟色胺间的关系。以吡喹酮 ( 5× 1 0 - 7mol/ L和 5× 1 0 - 8mol/ L )、硝硫氰醚 ( 1 0 - 4mol/ L和 1 0 - 5mol/ L )分别培养日本血吸虫成虫 4h和 8h ,反相离子对高效液相法测定其体内 5-HT的含量变化。结果显示 :吡喹酮和硝硫氰醚对日本血吸虫体内 5-HT等含量的影响与对照相比均无显著性差异。表明二者抗虫作用似不通过影响虫体 5-HT的生物合成和降解。     

Changes in the glutathione thiol-disulfide status in wheat grain by foliar sulphur fertilization: consequences for the rheological properties of dough

Storage proteins and glutathione in wheat play an important role in gluten network formation and can be modified by supplementation of nitrogen (N) and sulphur (S) in wheat plants. The glutathione thiol-disulfide status and its relationship to the molecular weight distribution wheat polymeric protein and dough rheological properties have been examined after different foliar S fertilizations (S derived from micronized elemental S and NS, a mixture of N urea and elemental S) applied at the post-anthesis stage. Changes in levels of reduced glutathione (GSH), glutathione disulfide (GSSG), polymeric protein-glutathione mixed disulfide (PPSSG) were analysed by reversed phase high performance liquid chromatography, during grain development using the wheat cultivars, Soissons and Trémie. During the grain desiccation phase, S supplementation (i) increased the GSSG/GSH ratio by 23–25% (ii) induced PPSSG accumulation, and (iii) decreased the formation of SDS-unextractable polymeric protein (UPP) and its molecular mass distribution. However, simultaneous N and S supplementation results in: (i) a decrease in PPSSG formation by 20–30% and (ii) an increase of UPP by 7–18% by enhancing both the branching of the aggregated proteins and their molecular weight. The mixograph parameters show that all forms of endogenous glutathione are linked to dough weakening and are negatively correlated with dough mixing tolerance, dough strength and consistency, while UPP is positively correlated with dough strength and consistency. These findings indicate that S nutrition influences dynamics of the glutathione forms in the grain and results in modification the degree of polymerization of storage protein. Thus both the changes in the form of glutathione and protein polymerization influence the rheological properties of dough.     

Molecular cloning, expression, and characterization of a phi-type glutathione S-transferase from Oryza sativa

The structural gene for glutathione S-transferase in Oryza sativa was successfully cloned from a cDNA library by the polymerase chain reaction method. The deduced amino acid sequence of this gene showed 44-66% similarity to the sequences of the class phi GSTs from Arabidopsis thaliana and Zea mays. This gene was expressed in Escherichia coli with the pET vector system and the gene product was purified to homogeneity by GSH-Sepharose affinity column chromatography. The expressed OsGSTF3-3 was a homo-dimer composed of 24 kDa subunit and its pI value was approximately 7.3. The OsGSTF3-3 was retained on GSH affinity column and its K_m value for GSH was 0.28 mM. The OsGSTF3-3 displayed high activity toward 1-chloro-2,4-dinitrobenzene, a general GST substrate and also had high activities towards acetanilide herbicides, alachlor, and metolachlor. The OsGSTF3-3 was highly sensitive to inhibition by benastatin A and S-hexyl-GSH. From these results, the expressed OsGSTF3-3 is a phi class GST and seems to play an important role in the conjugation of the chloroacetanilide herbicides.     

Se-containing spirulina phycocyanin attenuated liver injury induced by carbon tetrachloride in mice

AIM:To investigate the effect of Se-containing spirulina phycocyanin (Se-SPC) on liver injury of mice induced by carbon tetrachloride (CCl₄). METHODS:The mouse model was conducted by intragastric feeding with 2% CCl₄ oil for three times, meanwhile Se-SPC, spirulina phycocyanin (SPC) and Na₂SeO₃ were injected (ip) to various groups for 7 days. Then selenium (Se), glutathione peroxidase (GPx), superoxide dismutase (SOD), alanine aminotransferase (ALT), malondiaoldehyde (MDA) and nitric oxide (NO) levels in blood and liver were measured. RESULTS:The level of Se,GPx and SOD activities were obviously higher(P<0.05)but ALT activity,MDA and NO₂^-/NO₃^- levels were remarkably lower(P<0.05)in Se-SPC treated groups than those in CCl₄ groups,and effects of high dose Se-SPC on Se,GPx,MDA and NO₂^-/NO₃^- were even more significant(P<0.01).Under the same dose of Se or protein,effects of all selected targets in Se-SPC groups were more efficient than those in SPC groups and inorganic-Se groups.Furthermore,Se levels had a positive correlation with GPx activity(r=01705),which had negative correlation with levels of MDA,NO₂^-/NO₃^- and ALT(r=-0.629,r=-0.336,r=-0.457,respectively), and positive correlations between ALT activity and MDA or NO₂^-/NO₃^- level were found (r=0.519,r=0.641). CONCLUSION:These results indicated that Se-SPC may attenuate liver injury of mice induced by CCl₄ through its anti-inflammatory action and enhancing selenoenzyme expression.     

Effect of 4-chloro-2-methylphenoxyacetic acid and 2,4-dimethylphenol on human erythrocytes

The effects of exposure of human erythrocytes to different concentrations of 4-chloro-2-methylphenoxyacetic acid (MCPA) and its metabolite—2,4-dimethylphenol (2,4-DMP) were studied. The investigations concerned mainly the content of glutathione (GSH and GSSG), glutathione peroxidase (GSH-Px), glutathione transferase (GST), and the level of adenine energy charge (AEC). Reactive oxygen species (ROS) such as hydroxyl radical, superoxide anion, hydrogen peroxide, and nitric oxide are produced during normal processes in the cell. Under normal conditions, antioxidant systems of the cell minimize damage caused by ROS. When ROS generation increases to an extent that it overcomes the cellular antioxidant systems, the result is oxidative stress. We observed that MCPA and 2,4-DMP decreased the level of GSH in erythrocytes in comparison with control. MCPA did not affect glutathione peroxidase and glutathione transferase activity, while 2,4-DMP increased their activity. 2,4-DMP decreased the level of ATP and increased the content of ADP and AMP, leading to the fall of the level of AEC. MCPA and 2,4-DMP transform hemoglobin into methemoglobin, thus preventing oxygen transport. Comparison of the toxicity of MCPA and 2,4-DMP revealed that the most prominent changes occurred in human erythrocytes incubated with 2,4-DMP.     

日本大耳白兔抗氧化物酶活性增龄性变化研究

选择日本大耳白兔,按年龄分为幼年、青年、老年三组,分别测定动脉血血清SOD、MDA、GSH—px活性水平,以探讨日本大耳白兔抗氧化物酶活性增龄性变化。结果显示：随着年龄的增长,血清SOD活性在中年期以前是随增龄而增加,青年组显著高于幼年CR（P〈0．05,P〈0．01）,在中年期以后是随增龄而降低,老年组明显低于青年组（P〈0．05,P〈0．01）;血清MDA含量是随增龄而增加,老年组明显高于青年组和幼年组（P〈0．05,P〈0．01）,青年组明显高于幼年CR（P〈0．05）;血清GSH—px活性随增龄而降低,老年组显著低于青年组和幼年组（P〈0．01）,青年组显著低于幼年组（P〈0．05,P〈0．01）。机体在衰老过程中体内自由基生成增加。抗氧化物酶活性全面降低可能是加速机体衰老的原因之一。     

pVAX1/SjTSP2 DNA疫苗免疫预防小鼠日本血吸虫病的效果评估

为评估SjTSP2分子作为日本血吸虫DNA疫苗候选抗原分子的潜力,本实验扩增了编码该分子的DNA片段,构建了重组真核质粒pVAX1/SjTSP2,检测其在哺乳动物细胞中表达情况后,通过基因枪轰击法免疫小鼠,应用ELISA、流式细胞术等检测重组质粒诱导的免疫反应,计算减虫减卵率,评估对小鼠日本血吸虫病的免疫保护效果。间接免疫荧光试验结果显示该质粒能在293T细胞中表达。质粒免疫小鼠诱导了显著升高的特异性IgG抗体和IFN-γ、IL-4等细胞因子。小鼠免疫保护试验结果表明,免疫组小鼠虫荷数较PBS对照组增加12.4%(P=0.368),但肝组织虫卵减少10.01%(P=0.486)。说明制备的DNA疫苗能刺激小鼠产生较强的免疫反应,但诱导的免疫保护作用并不理想,为血吸虫DNA疫苗研究提供了参考数据。     

自然感染日本血吸虫病耕牛血清循环抗原消长规律的研究

为了观察疫区自然感染日本血吸虫病耕牛血清循环抗原消长规律,本试验将血吸虫病非疫区的水、黄牛各１０头,转运至安徽省血吸虫病疫区,接受为期８周的自然感染。用斑点酶联免疫吸附试验测定牛血清循环抗原。结果,水、黄牛血清循环抗原滴度均于自然感染第４周后明显上升,水、黄牛自然感染第８周的血清循环抗原最高滴度分为１∶１０２４０和１∶５１２０,最低滴度则分别为１∶１２８０和１∶６４０,水牛循环抗原滴度高于黄牛。虫荷数与血清循环抗原滴度未见有相关性。     

日本血吸虫谷胱甘肽S－转移酶小鼠免疫试验

用谷胱甘肽琼脂糖亲和层析柱纯化中国大陆株日本血吸虫成虫谷胱甘肽Ｓ－转移酶（ＧＳＴ）。把纯化的ＧＳＴ结合福氏性剂免疫了二批达ＢＡＬＢ／ｃ小鼠，获得了２９．５８～３２．７１％的减虫率和５２．９４～６８．１３％的粪便减卵率。ＥＬＩＳＡ试验表明免疫小鼠产生了体液免疫应答，免疫印迹试验（ＷｅｓｔｅｒｎＢｌｏｔｔｉｎｇ）显示日本血吸虫水溶性成虫抗原和ＧＳＴ抗原的２６、２８ｋＤ蛋白被免疫小鼠血清所识别。     

氨氮胁迫对四鼻须鲤鱼肝脏GST酶活力的影响

实验采用生态毒理学方法,以NH4Cl为实验药物,设置0、50、150和250mg/L实验浓度。通过对野生和养殖群体的四鼻须鲤鱼氨氮对肝脏谷胱甘肽巯基转移酶（GST）活力的影响,比较两个群体抗氨氮能力。结果表明：在相同的氨氮浓度和胁迫时间下,四鼻须鲤鱼养殖群体GST酶活力的变化趋势与野生群体大致相同。除250mg/L浓度组在胁迫5d和10d时,野生群体酶活性略低于养殖群体外,在其余时间点的野生群体酶活性均高于养殖群体,而且,在高浓度氨氮胁迫下,野生群体肝脏的GST酶活性达到峰值的时间比养殖群体更短,显示出较强的抗氨氮能力。