肾上腺素受体和乙酰胆碱受体在血吸虫性肝纤维化肝脏组织中的表达研究

目的研究α1A-肾上腺素受体(α1A-AR)和M1型乙酰胆碱受体(mAChRM1)在日本血吸虫性肝纤维化小鼠肝脏组织中的表达情况,初步探讨神经递质受体在肝纤维化发生机制中的作用。方法清洁级昆明小鼠30只随机分为正常组(N组,10只)、模型组(M组,20只),模型组小鼠用腹部敷贴尾蚴法感染日本血吸虫制备肝纤维化模型。应用免疫组织化学、逆转录聚合酶链反应(RT-PCR)和免疫印迹(Western blot)等方法检测肝脏组织中α1A-AR和mAChRM1表达变化。结果模型组肝脏组织中α1A-AR和mAChRM1表达较正常组明显增加,其差异具有显著性统计学意义(P<0.05)。结论神经递质受体表达上调在血吸虫性肝纤维化发生发展中起一定作用。     

稻螟赤眼蜂对二化螟和台湾稻螟的控制潜能评价

为了明确稻螟赤眼蜂种群增长力与水稻螟虫种群增长力之间的关系和评价稻螟赤眼蜂的控害能力,通过生命表方法,组建了二化螟和台湾稻螟在水稻上的实
验种群生命表,以及稻螟赤眼蜂在二化螟卵和台湾稻螟卵上的生殖力表,测定了稻螟赤眼蜂对两种螟虫卵的寄生能力.结果表明：二化螟和台湾稻螟的世代历期（T）分别为56.40 d和47.80 d,内禀增长率（r_m）分别为0.0489和0.072.稻螟赤眼蜂在二化螟卵和台湾稻螟卵上的平均T分别为9.75 d和9.78 d,r_m分别为0.3161和0.3154.通过比较分析稻螟赤眼蜂与两种螟虫的实验种群生命表参数,可知稻螟赤眼蜂能够有效控制两种水稻螟虫,赤眼蜂种群增长力相对于螟虫种群是超前而不是跟随关系.     

大孔树脂吸附法分离海金沙总黄酮的研究

用大孔树脂吸附法分离海金沙中的总黄酮,考查了解吸剂浓度、吸附固液比(g/mL)、吸附温度和吸附时间对海金沙总黄酮提取率的影响。结果表明:用70%乙醇作解吸剂,在吸附固液比为1∶6(g/mL),50℃温度下吸附90 min,总黄酮提取率为1.81%。     

海金沙草总黄酮提取工艺的响应面优化

为了提高海金沙草总黄酮提取效率,运用了Plackett-Burman试验设计、爬陡坡试验和Box-Behnken设计对提取工艺进行了响应面优化试验。利用Plackett-Burman对影响总黄酮提取的诸多相关因素进行了评价并成功筛选出主效应因子,即提取时间、提取温度和乙醇浓度。在Plackett-Burman设计基础上,根据主效应因子作用大小与方向进行了爬陡坡试验。最后用Box-Behnken响应面技术优化了总黄酮提取工艺并建立了关键影响产量的二次多项式数学模型,解模型逆矩阵得最优解(优化方案),即提取温度X1=45.45℃,乙醇体积分数X2=47.1%、提取时间X3=84.8 min。模型预测结果为0.433 mg.L-1,验证试验结果为0.428±0.004 mg.L-1(n=3)。     

Histamine signalling in Schistosoma mansoni: Immunolocalisation and characterisation of a new histamine-responsive receptor (SmGPR-2)

In parasitic platyhelminthes, including Schistosoma mansoni, biogenic amines play several important roles in the control of motility, metabolism and reproduction. A bioinformatics analysis of the S. mansoni genome identified approximately 16 full-length G protein-coupled receptors (GPCRs) that share significant homology with aminergic receptors from other species. Six of these sequences are structurally related to SmGPR-1 (formerly SmGPCR), a previously described histamine receptor of S. mansoni, and constitute a new clade of amine-like GPCRs. Here we report the cloning of a second member of this clade, named SmGPR-2. The full-length receptor cDNA was expressed in Saccharomyces cerevisiae and shown to be activated by histamine and 1-methylhistamine, whereas other common biogenic amines had no significant effect. Antagonist assays showed that SmGPR-2 was inhibited by classical biogenic amine antagonists but the pharmacological profile was unlike those of known mammalian histamine receptors. Confocal immunolocalisation studies revealed that SmGPR-2 was expressed in the nervous system and was particularly enriched in the subtegumental neuronal plexus of adult S. mansoni and larvae. The ligand, histamine, was found to be widely distributed, mainly in the peripheral nervous system including the subtegumental plexus where the receptor is also expressed. Finally, SmGPR-2 was shown to be developmentally regulated at the RNA level. Quantitative PCR studies showed it was up-regulated in the parasitic stages compared with cercaria and expressed at the highest level in young schistosomula. The widespread distribution of histamine and the presence of at least two receptors in S. mansoni suggest that this transmitter is an important neuroactive substance in schistosomes.     

Imatinib has a fatal impact on morphology, pairing stability and survival of adult Schistosoma mansoni in vitro

Schistosomes cause bilharzia (schistosomiasis), one of the most prevalent parasitic diseases for human and animals worldwide. Praziquantel (PZQ) is the only widely used drug for treatment and control of this parasitemia. Since a vaccine is not yet available, and in light of emerging resistance against PZQ, the search for alternatives has high priority. Here we present that Imatinib, a compound used in human cancer therapy (Gleevec; STI-571), significantly affected schistosome morphology and physiology in vitro. Besides its negative effect on gonad development and pairing stability, Imatinib led to pathological alterations of the gastrodermis, which finally caused the death of the parasite.     

Mapping the Catalytic Cycle of Schistosoma mansoni Thioredoxin Glutathione Reductase by X-ray Crystallography

Schistosomiasis is the second most widespread human parasitic disease. It is principally treated with one drug, praziquantel, that is administered to 100 million people each year; less sensitive strains of schistosomes are emerging. One of the most appealing drug targets against schistosomiasis is thioredoxin glutathione reductase (TGR). This natural chimeric enzyme is a peculiar fusion of a glutaredoxin domain with a thioredoxin selenocysteine (U)-containing reductase domain. Selenocysteine is located on a flexible C-terminal arm that is usually disordered in the available structures of the protein and is essential for the full catalytic activity of TGR. In this study, we dissect the catalytic cycle of Schistosoma mansoni TGR by structural and functional analysis of the U597C mutant. The crystallographic data presented herein include the following: the oxidized form (at 1.9 Å resolution); the NADPH- and GSH-bound forms (2.3 and 1.9 Å, respectively); and a different crystal form of the (partially) reduced enzyme (3.1 Å), showing the physiological dimer and the entire C terminus of one subunit. Whenever possible, we determined the rate constants for the interconversion between the different oxidation states of TGR by kinetic methods. By combining the crystallographic analysis with computer modeling, we were able to throw further light on the mechanism of action of S. mansoni TGR. In particular, we hereby propose the putative functionally relevant conformational change of the C terminus after the transfer of reducing equivalents from NADPH to the redox sites of the enzyme.     

Schistosoma japonicum: The design and experimental evaluation of a multivalent DNA vaccine

The aim of this study was to construct and evaluate the immunity efficacy of the DNA multivalent vaccine pVIVO₂SjFABP-23. The vaccine was constructed and produced as follows. Forty BALB/c mice were divided into four groups designated pVIVO₂, pVIVO₂Sj23, pVIVO₂SjFABP and pVIVO₂SjFABP-23. Each mouse was immunized with 100 μg of the corresponding plasmid DNA by intramuscular injection. 28 days post-vaccination, the mice were challenged with S. japonicum cercariae, and the worm and egg burdens were determined 42 days post-challenge. Serum samples were collected from all the mice before and after vaccination and at the end of the experiment, and used for antibody detection. The IFN-γ and IL-4 levels were quantified in the supernatants of specifically stimulated spleen cells. The number of worms was reduced by 52%, 40% and 42% in mice respectively immunized with pVIVO₂SjFABP-23, pVIVO₂Sj23 or pVIVO₂SjFABP. A respective 61%, 38% and 39% egg reduction was determined relative to those mice that only received the empty pVIVO2 plasmid. pVIVO₂SjFABP-23 immunization increased IgG levels against SWAP and SEA. Increased IFN-γ levels were detected in the supernatant of specific stimulated spleen cells from mice immunized with the 3 different constructs. The multivalent DNA vaccine developed induced higher levels of protection than the two monovalent tested vaccines.     

Photosynthetic rate and lectin activity of soybean leaves after inoculation with rhizobia together with homologous lectin

Nodule bacteria (Bradyrhizobium japonicum) of various activities were preincubated with homologous lectin and then used for inoculating soybean (Glycine max (L.) Merrill) seeds. The effect of this inoculation on the photosynthetic rate, lectin activity in leaves, and plant development at different supply of mineral nitrogen was investigated under the conditions of pot experiments. There was a positive relationship between the photosynthetic rate and the lectin activity of proteins isolated from soybean leaves. Under the conditions of effective symbiosis, activation of functioning of the symbiotic apparatus by preincubation of the rhizobia with lectin exerted an additional stimulating effect on the photosynthetic rate. It is suggested that a relationship between the effectiveness of legume-rhizobium symbiosis and the lectin activity in leaves is mediated by the regulation of photosynthesis through a demand for assimilates in the source-sink system of soybean plants.