Enhanced expression of heterologous inulinase in <Emphasis Type="Italic">Kluyveromyces lactis</Emphasis> by disruption of <Emphasis Type="Italic">hap1</Emphasis> gene

Inulinase gene (Kcinu) derived from Kluyveromyces cicerisporus was expressed extracellularly in Kluyveromyces lactis using an episomal vector directed by Kcinu promoter. The influence of hap1 gene disruption on the expression of inulinase was studied. Inulinase activity in the supernatant of the recombinant Klhap1Δ strain was 391 U ml⁻¹ after cultured 120 h, which was 2.2-fold that of the wild type host. The relative inulinase mRNA level of the Klhap1Δ strain was 11.3-fold that of the wild type strain, and the expression plasmid was more stable in the mutant host. Based on these results, the disruption of hap1 facilitated the high and stable expression of inulinase controlled by Kcinu promoter in K. lactis.     

北方常见农业土地利用方式对土壤螨群落结构的影响

土壤螨群落与土壤健康状况密切相关,农业土地利用方式会影响土壤螨的生存环境,进而影响螨的群落分布。在北方传统农业区研究了小麦玉米常规轮作农田,1年温室蔬菜大棚、4年温室蔬菜大棚和4年露天菜地下土壤主要理化指标和螨群落结构。研究结果表明露天菜地和温室大棚的利用方式均会提高土壤碱解氮和有机质含量,同时温室大棚的长期利用会增加土壤有效磷含量。螨群落分析显示常规农田中甲螨亚目为优势类群;常规农田转变为温室大棚后,由于施肥造成土壤营养物质含量增加,以及人类对土壤扰动程度的提高,无气门亚目取代甲螨亚目成为温室内螨优势类群。但随着温室年限的延长,螨多样性和粉螨科丰度都要有所下降,这可能是磷累积和强扰动效应的共同后果。常规农田转变为露天菜地后,加强的人类扰动也会降低甲螨亚目丰度,但与温室螨群落相比,4个亚目分布要相对均匀。温室在3种土地利用类型中对土壤螨的负面影响最为明显,从土壤功能的自我维持和修复方面来讲是十分不利的,温室内土壤生物多样性的保护尤其值得人们关注和重视。     

有机铬对实验性糖尿病大鼠胰岛β细胞形态结构及功能的影响

目的探讨有机铬(2-吡啶甲酸铬)对大鼠胰岛β细胞形态结构及内分泌功能的影响。方法首先通过尾静脉注射新鲜配制的1.5%四氧嘧啶溶液制备糖尿病大鼠模型;通过灌胃的方式提高糖尿病大鼠体内有机铬含量;治疗12周后,通过氧化酶法测定大鼠血清葡萄糖水平,利用免疫组织化学方法和放射免疫学方法分别观察大鼠胰岛β细胞形态结构的改变及大鼠血清胰岛素含量的变化。结果两治疗组大鼠血清葡萄糖水平明显下降,与糖尿病模型组相比,差异均有显著性;400μg治疗组大鼠体内胰岛素水平明显升高,与糖尿病模型组相比差异有显著性,而200μg治疗组大鼠血清胰岛素水平虽有所增加但与糖尿病组相比,差异无显著性;免疫组化显示治疗组大鼠胰岛β细胞数量增多,胞质内胰岛素免疫反应阳性颗粒明显增多。结论有机铬对糖尿病大鼠具有明显的降低血糖功能,并能促进受损胰岛及β细胞形态结构功能的恢复,对糖尿病大鼠病理状态具有明显的改善作用。     

热环境下复合电解质饮料的生理生化效应

本实验观察了复合电解质饮料对热环境下大白鼠与人体的生理生化效应。实验结果表明,复合电解质饮料具有维持人体在热环境下劳动时的水与电解质平衡,保留水份在体内,减少水份消耗,改善细胞能量代谢,提供能量,消除疲劳和提高耐热能力等作用。     

Efficient screening and breeding of Bacillus thuringiensis subsp. kurstaki for high toxicity against Spodoptera exigua and Heliothis armigera

Spodoptera exigua is one of the most renowned agricultural pest insects and relatively insensitive to Bacillus thuringiensis subsp. kurstaki strains which are widely used commercial products to control lepidopterans such as Heliothis armigera. In the current study, we have developed a new and efficient approach to screen and breed a B. thuringiensis subsp. kurstaki strain exhibiting high toxicity against S. exigua while retaining its high toxicity against H. armigera. UV and diethyl sulfate methods were used for mutagenesis, followed by an agar plug plate diffusion assay for preliminary screening of Zwittermicin A over-producing mutants, from which we obtained a mutant strain, designated here as B. thuringiensis subsp. kurstaki D1-23, with high toxicity against S. exigua. The toxicity of D1-23 against S. exigua and H. armigera was improved by 115.4 and 25.9%, respectively, compared to its parental commercial strain BMB005.     

应用系统聚类分析的方法评价珠江流域北江水系的水质状况

硅藻被认为是水体环境的良好指示藻类。本文利用着生硅藻结合水体中的物理、化学等因素,以系统聚类分析方法对珠江流域北江水系的8条河流的水质状况进行评价,证明这些河流的水质是清洁的。     

镉污染对不同生境拟水狼蛛氧化酶和金属硫蛋白应激的影响

为探讨镉(Cd)对机体抗氧化功能及金属硫蛋白(MT)的影响,在室内分别用不添加Cd2 和添加浓度为20 mgkg-1 Cd2 培养基培养的黑腹果蝇来饲喂4种不同生境下(S1,S2,S3和S4)拟水狼蛛,于饲喂5d、10d和20d后,分别测定其体内MT和丙二醛 (MDA)的含量及超抗氧化酶(GST、SOD和CAT)活性。结果表明：1,不同生境拟水狼蛛用不添加Cd2 培养基培养的黑腹果蝇饲喂后,不添加Cd2 对照组拟水狼蛛镉的积累量和MT含量无显著变化,但均显著低于添加Cd2 污染组。添加Cd2 污染组拟水狼蛛镉的积累量和MT含量都显著高于对照组,且均随着饲喂时间的延长而显著升高,具有明显的时间–效应关系(p<0.05)。2,在饲喂5d和10d后,不添加Cd饲喂的拟水狼蛛MDA含量和抗氧化酶系差异都不显著。添加Cd2 污染组(S1,S2和S3)MDA含量显著高于对照组(S4),MDA含量与饲喂时间呈显著正相关(p<0.05);GST、SOD和CAT等抗氧化酶活性污染组显著低于对照组,与饲喂时间呈显著负相关(p<0.05);饲喂20d后,污染组MDA含量和抗氧化酶（SOD和CAT）活性均与对照组无显著差异,但GST活性差异显著。     

NeuronGrowth, a software for automatic quantification of neurite and filopodial dynamics from time-lapse sequences of digital images

We developed NeuronGrowth, a software for the automatic quantification of extension and retraction of neurites and filopodia, from time-lapse sequences of two-dimensional digital micrographs. NeuronGrowth requires a semiautomatic characterization of individual neurites in a reference frame, which is then used for automatic tracking and measurement of every neurite over the whole image sequence. Modules for sequence alignment, background subtraction, flat field correction, light normalization, and cropping have been integrated to improve the quality of the analysis. Moreover, NeuronGrowth incorporates a deconvolution filter that corrects the shadow-cast effect of differential interference contrast (DIC) images. NeuronGrowth was tested by analyzing the formation of outgrowth patterns by individual leech neurons cultured under two different conditions. Phase contrast images were obtained from neurons plated on CNS homogenates and DIC images were obtained from similar neurons plated on ganglion capsules as substrates. Filopodia were measured from fluorescent growth-cones of chick dorsal root ganglion cells. Quantitative data of neurite extension and retraction obtained by three different users applying NeuronGrowth and two other manually operated software packages were similar. However, NeuronGrowth required less user participation and had a better time performance when compared with the other software packages. NeuronGrowth may be used in general to quantify the dynamics of tubular structures such as blood vessels. NeuronGrowth is a free plug-in for the free software ImageJ and can be downloaded along with a user manual, a troubleshooting section and other information required for its use from http://www.ifc.unam.mx or http://www.ifc.unam.mx/ffm/index.html.