辣椒素对大鼠肠系膜下神经诱发动作电位的影响

目的:探讨辣椒素对肠系膜下神经诱发动作电位的影响。方法:对大鼠肠系膜下神经节施加不同浓度(分别为2.5、5、10g/L)的辣椒素或其载体(对照)后,在其中枢端给予能引起反应的方波刺激,记录节后神经外周端的动作电位。结果:虽然实验动物存在个体差异,且辣椒素对其神经作用的阈值也有所不同,但在大多数情况下,当辣椒素浓度为2.5～5g/L、作用3min后,即可使神经的敏感性降低;并表现出了较为明显的剂量相关性,即随着辣椒素浓度的升高,其对神经的脱敏作用也逐渐增强。结论:辣椒素对肠系膜下神经节内的交感和副交感神经均具有抑制作用。     

Prostaglandin E2 inhibits TNF production in murine bone marrow-derived dendritic cells

Exposure to pathogens induces dendritic cells to release inflammatory cytokines and chemokines. The inflammatory response is controlled by endogenous agents such as anti-inflammatory cytokines, glucocorticoids, anti-inflammatory neuropeptides, and lipid mediators. This study is the first report on the inhibition by prostaglandin E2 (PGE2) of TNF release from bone marrow-derived dendritic cells stimulated with lipopolysaccharide (LPS), a TLR4 ligand, or peptidoglycan, a TLR2 ligand. The inhibition of TNF occurs at both mRNA and protein level. The inhibitory effect of PGE2 is mediated by the EP2 and EP4 receptors, and involves both PKA signaling and mediation by DC-derived IL-10. Intraperitoneal administration of PGE2 together with LPS results in a reduction in serum TNF and intracellular TNF in peritoneal exudate cells, compared to LPS alone. In addition, administration of PGE2 in vivo reduces the numbers of CD11c+ DCc that accumulate in the peritoneal cavity in response to LPS. The various implications of the PGE2-induced reduction in TNF are discussed.     

迷迭香植物精油对白纹伊蚊的驱避作用及其化学成分

应用水蒸气蒸溜法和超临界二氧化碳萃取法提取生长于云南的迷迭香茎叶植物精油,测试2种精油对白纹伊蚊Aedes albopictus Skuse的驱避作用,并利用GC/MS联用仪分析2种精油的化学成分。结果表明:超临界二氧化碳提取的迷迭香精油对白纹伊蚊的驱避时间是(5.82±1.13)h,达到国标驱避剂评价的B级标准。2种精油化学成分中都含有驱纹活性物质:柠檬烯、樟脑、Eucalyptol、(+)-4-Carene、1-methyl-4-(1-methyle thylidene)-cyclohexene、Beta-pinene等,且驱蚊活性成分在挥发油中占有较高比例,显示该植物精油在驱蚊方面有一定开发价值。     

小麦秆锈抗性遗传及抗性基因研究进展

目前国际上已发现近80个小麦抗秆锈基因,其中45个抗秆锈基因已被正式定名,58个抗秆锈基因已定位在小麦特定染色体上,其中12个基因被标记。本文对小麦抗秆锈病基因抗源、抗秆锈性遗传、分子标记研究现状及存在问题加以综述,并对抗秆锈分子遗传前景进行展望。     

"油脂酸价的测定"实验中若干问题的探讨

通过油脂酸价测定的预实验,发现了一些应注意的问题。滴定油脂所用氢氧化钾溶液的浓度应根据油脂酸败的程度来确定,实验应该考虑所用油脂的种类,实验结果平行数之间的误差不应太大。提示应充分重视学生实验的预实验。     

体外诱导骨髓间充质干细胞向肝细胞样细胞方向分化

探讨成纤维生长因子-2(FGF-2)在体外定向诱导大鼠骨髓间充质干细胞(BM-MSCs)向肝细胞样细胞分化的作用及量化关系。体外分离培养大鼠BM-MSCs,将第3代BM-MSCs采用不同剂量的FGF-2诱导。诱导后,在显微镜下观察细胞形态学的改变;用免疫细胞化学法检测白蛋白和CK19的分泌;Shiff染色法检测糖原的分泌。诱导后BM-MSCs由梭形向多角形、卵圆形方向变化,白蛋白、CK19和糖原12 d即有阳性表达,以后随着诱导时间的延长阳性率逐渐升高。20 ng/mL FGF-2诱导比10 ng/mL FGF-2诱导细胞白蛋白、CK19和糖原的表达量均多。20 ng/mL FGF-2具有较强的诱导BM-MSCs向肝细胞样细胞分化的能力。     

Human umbilical cord-derived MSC culture: the replacement of animal sera with human cord blood plasma

Human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) hold great potential for their therapeutic use in various clinical diseases. Many publications have reported on human blood-derived alternatives to animal serum for culturing mesenchymal stem cells, such as human serum, allogenic umbilical cord blood serum, and human platelet derivatives. However, it is not clear whether human umbilical cord blood plasma (UCBP), as the surplusage of umbilical cord blood mesenchymal stem cell extraction, could be used. In this study, in order to make the best of umbilical cord blood, the human UCBP was dialyzed to replace fetal bovine serum (FBS) in the culture medium. hUC-MSCs were cultured in the new medium. Cell growth rate, specific biomarkers, and differentiation properties were detected to characterize the cell proliferation and MSC-specific properties. The hUC-MSCs cultured in such derived medium were verified with proliferation rate, cluster differentiation markers, cell cycle, as well as differentiation capabilities. Such dialyzed human UCBP is fully comparable with, if not superior to, FBS in deriving and culturing hUC-MSCs.     

Disruption of YPS1 and PEP4 genes reduces proteolytic degradation of secreted HSA/PTH in Pichia pastoris GS115

Human serum albumin (HSA) and human parathyroid hormone (1-34) [PTH (1-34)] fusion protein [HSA/PTH (1-34)] is a promising long-acting form of PTH (1-34) for osteoporosis treatment. Secretory expression of intact HSA/PTH (1-34) in Pichia pastoris GS115 was accompanied by two degradation fragments, with molecular weights around 66 kDa, in addition to the well-known ~45 kDa HSA-truncated fragment, resulting in a low yield of intact protein. In this study, two internal cleavage sites were identified in the PTH (1-34) portion of the fusion protein by Western Blot analysis. To minimize proteolytic cleavages, several protease genes including PEP4 (encoding proteinase A), PRB1 (proteinase B) and seven YPSs genes (yapsin family members) were knocked out respectively by disruption of the individual genes and the selective combinations. Reduced degradation was observed by single disruption of either PEP4 gene or YPS1 gene, and the lowest level of degradation was observed in a pep4△yps1△ double disruptant. After 72 h of induction, more than 80 % of the HSA/PTH (1-34) secreted by the pep4△yps1△ double disruptant remained intact, in comparison to only 30 % with the wild-type strain.     

全氟辛烷磺酰基化合物对剑尾鱼肝脏抗氧化物酶活性的影响

目的研究全氟辛烷磺酰基化合物(PFOS)暴露对剑尾鱼(Xiphophorus helleri Heckel)抗氧化物酶活性的影响,探讨PFOS对鱼类的致毒机理。方法使用浸润法以3.5、7.0、14.0和28.0 mg/L四个PFOS浓度为剑尾鱼染毒,定量测定了96 h内肝脏组织中的超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-PX)活性及丙二醛(MDA)含量的变化。结果 PFOS暴露12 h后,除28.0 mg/L组SOD活性被显著性抑制外,其余各组与对照组均无显著性差异(P>0.05);7.0 mg/L组和14.0 mg/L组在24 h被极显著诱导(P<0.01),并且一直保持至96 h。CAT活性随PFOS浓度的升高而降低,12 h时,除3.5 mg/L组外,其余各组CAT活性被显著或极显著抑制,至24 h时,各组CAT活性有上升趋势,但48 h后,各组呈不断下降趋势持续至96 h,其CAT活性恢复到12 h水平。GSH-PX活性变化与CAT活性变化趋势相似,其中28.0 mg/L组在不同时间均被显著性抑制,并在96 h时抑制率达到最高值64.8%。MDA含量在12 h时呈小幅下降趋势,但随着暴露时间的延长,各处理组MDA含量呈连续上升趋势,并在96 h时达到最高点,诱导率分别为71.2%、70.1%和85.1%。结论结果表明,SOD的高活性是由于机体中超氧阴离子的存在,而高浓度的超氧阴离子能够灭活CAT和GSH-PX活性,因此,CAT和GSH-PX活性始终低于对照组。GSH-PX对PFOS的敏感性高于CAT。MDA含量持续升高反映出细胞组织已经遭受到氧化损伤。剑尾鱼活体的实验表明,PFOS能够诱导肝脏氧化应激反应,氧化损伤是PFOS致毒的主要途径之一。     

Proteomic Analysis of Interactions Between the Generalist Herbivore <Emphasis Type="Italic">Spodoptera exigua</Emphasis> (Lepidoptera: Noctuidae) and <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

In this study, comparative proteomics was used to investigate the interaction of Spodoptera exigua and Arabidopsis thaliana. By using 2-D electrophoresis of differentially expressed proteins, combined with high-throughput matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) and MALDI-TOF/TOF MS, the changes in the abundance of proteins induced by insect feeding were studied in A. thaliana. More than 1,100 protein spots were reproducibly detected on each gel. The intensities of 30 protein spots in particular changed significantly, showing differences in volume of at least twofold. Among these, 17 protein spots were upregulated, and 13 were downregulated following an 8-h insect feeding period. Nineteen insect-feeding-responsive proteins were identified, all of which were involved in metabolic regulation, binding functions or cofactor requirement of protein, cell rescue, and defense and virulence, as assessed by Munich Information Center for Protein Sequences function category. About 50% of these were involved in metabolism, including transketolase, S-adenosylmethionine synthase 3, 2,3-biphosphoglycerate-independent phosphoglycerate mutase, beta-ureidopropionase, GDP-d-mannose 3′,5′-epimerase, and fatty acid synthase. The identification of insect-feeding-responsive proteins on Arabidopsis provides not only new insights into insect stress but also a good start for further investigation of their functions. Understanding how the plant responses to insects in the proteomic level will provide tools for a better management of insect pest in the field.