猪流感病毒HA基因的克隆及杆状病毒转移载体的构建

猪流感病毒(swine influenza virus, SIV)不仅给养猪业造成重大经济损失,而且威胁人类健康.血凝素(hemagglutinin,HA)是SIV囊膜上的重要免疫原,介导病毒吸附和膜融合.针对流感病毒的中和抗体主要以HA靶标阻断受体结合,本研究通过RT-PCR扩增了猪流感病毒A/Swine/Inner Mongolia/547/01(H3N2)的HA基因,并将其克隆入pMD18-T载体.重组pMD18-T经Kpn I 和 Pst I 酶切后得到的HA基因插入pMelBacA载体的Kpn I/Pst I位点.PCR和限制性内切酶分析鉴定表明成功构建了pMelBacA-HA转移载体.这为开发HA亚单位疫苗奠定了基础.     

A new <Emphasis Type="Italic">Dactylella</Emphasis> species from <Emphasis Type="Italic">Orbilia alba</Emphasis>

A new Dactylella species, Dactylella alba was isolated from the ascospores of Orbilia alba collected in Wenshan County, Yunnan Province, China. Conidiophores were either not branched or occasionally branched, bearing divergent sterigmata on the tip with single conidium on each. Conidia were elongated ellipsoids, 1–2 septate, mostly 1 septate. By combining the ITS sequence with morphological characteristics, a new anamorphic species is described and illustrated together with its teleomorph.     

Free radical scavenging activity of Lactobacillus fermentum in vitro and its antioxidative effect on growing–finishing pigs

Aims:  To evaluate the free radical-scavenging capacity of Lactobacillus fermentum and its effects on antioxidant enzyme levels in finishing pigs.
Methods and Results:  The free radical-scavenging activity of Lact. fermentum was analysed in vitro . The tested Lactobacillus showed a high scavenging ability against DPPH (1,1-diphenyl-2-picrylhydrazyl), superoxide and hydroxyl radicals which was dose dependent. Subsequently, 108 crossbred pigs weighing 20·67 BW, were allotted to dietary treatments including a basal diet or the basal diet supplemented with either aureomycin or 10·2 × 10⁷  Lact. fermentum CFU g⁻¹ diet. Supplementation of Lact. fermentum increased total antioxidant capacity ( P  < 0·01) in serum from 50 kg pigs, while serum superoxide dismutase ( P  = 0·01) and glutathione peroxidase ( P  < 0·01) increased, and malondialdehyde levels decreased ( P  < 0·01) in 90 kg pigs. Hepatic catalase ( P  = 0·04), muscle superoxide dismutase ( P  < 0·01) and copper–zinc-superoxide dismutase were enhanced ( P  = 0·01), whereas malondialdehyde levels were reduced ( P  = 0·05) by Lact. fermentum .
Conclusions:  The free radical-scavenging capacity of Lact. fermentum was dose dependent and its supplementation improved the antioxidant status of pigs.
Significance and Impact of the Study:  Lactobacillus fermentum could be used to alleviate oxidative stress and increase pig performance and improve pork quality.     

Comparative metabolomic analysis on industrial continuous and batch ethanol fermentation processes by GC-TOF-MS

The intracellular metabolic profile characterization of Saccharomyces cerevisiae throughout industrial ethanol fermentation was investigated using gas chromatography coupled to time-of-flight mass spectrometry. A total of 143 and 128 intracellular metabolites in S. cerevisiae were detected and quantified in continuous and batch fermentations, respectively. The two fermentation processes were both clearly distinguished into three main phases by principal components analysis. Furthermore, the levels of some metabolites involved in central carbon metabolism varied significantly throughout both processes. Glycerol and phosphoric acid were principally responsible for discriminating seed, main and final phases of continuous fermentation, while lactic acid and glycerol contributed mostly to telling different phases of batch fermentation. In addition, the levels of some amino acids such as glycine varied significantly during both processes. These findings provide new insights into the metabolomic characteristics during industrial ethanol fermentation processes. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

西双版纳自然保护区9种植被亚型象甲科多样性比较

2004年3–4月对西双版纳自然保护区季节性雨林、山地雨林、半常绿季雨林、落叶季雨林、石灰岩山季雨林、季风常绿阔叶林、暖温性落叶阔叶林、暖热性松林、热性竹林等9种植被亚型中象甲科昆虫多样性进行了研究。运用震落法采集标本853号,隶属于18亚科52属96种。跳象亚科个体数量最多,隐喙象亚科种类最丰富。季节性雨林和半常绿季雨林中不仅物种数最丰富,狭适种和稀有种也最多。暖温性落叶阔叶林及季节性雨林个体数量最丰富,落叶季雨林及半常绿季雨林最贫乏。9种植被亚型中象甲群落极不相似。季节性雨林和半常绿季雨林在生物多样性保护上具有重要地位,应予以优先考虑。     

Snapin-regulated late endosomal transport is critical for efficient autophagy-lysosomal function in neurons

Neuron maintenance and survival require late endocytic transport from distal processes to the soma where lysosomes are predominantly localized. Here, we report a role for Snapin in attaching dynein to late endosomes through its intermediate chain (DIC). snapin(-/-) neurons exhibit aberrant accumulation of immature lysosomes, clustering and impaired retrograde transport of late endosomes along processes, reduced lysosomal proteolysis due to impaired delivery of internalized proteins and hydrolase precursors from late endosomes to lysosomes, and impaired clearance of autolysosomes, combined with reduced neuron viability and neurodegeneration. The phenotypes are rescued by expressing the snapin transgene, but not the DIC-binding-defective Snapin-L99K mutant. Snapin overexpression in wild-type neurons enhances late endocytic transport and lysosomal function, whereas expressing the mutant defective in Snapin-DIC coupling shows a dominant-negative effect. Altogether, our study highlights new mechanistic insights into how Snapin-DIC coordinates retrograde transport and late endosomal-lysosomal trafficking critical for autophagy-lysosomal function, and thus neuronal homeostasis.     

Development and validation of a rapid capillary zone electrophoresis method for the determination of aconite alkaloids in aconite roots

Introduction – Aconites, with aconite alkaloids as the major therapeutic and toxic components, are used for the treatment of analgesic, antirheumatic and neurological symptoms. Quantification of the aconite alkaloids is important for the quality control of aconite‐containing drugs. Objective – To establish a validated capillary zone electrophoresis (CZE) method for the simultaneous determination of six major alkaloids, namely aconitine, mesaconitine, hypaconitine, benzoylaconine, benzoylmesaconine and benzoylhypaconine, in crude and processed aconite roots. Methodology – The CZE method was optimised and validated using a stability‐indicating method. The optimised running buffer was a mixture of 200 mm Tris, 150 mm perchloric acid and 40% 1,4‐dioxane (pH 7.8) with the capillary thermostated at 25°C. Results – Using the optimised method, six aconite alkaloids were well separated. The established method showed good precision, accuracy and recovery. Contents of these alkaloids in crude and processed aconites were determined and it was observed that the levels of individual alkaloids varied between samples. Conclusion – The developed CZE method was reliable for the quality control of aconites contained in herbal medicines. The method could also be used as an approach for toxicological studies. Copyright © 2009 John Wiley & Sons, Ltd.     

CCR2-64I allele is associated with the progression of AIDS in a Han Chinese population

It has been reported that the CCR5-Delta32, CCR2-64I and SDF1-3′A polymorphisms have protective effects against HIV-1 infection and can delay the progression of AIDS in European populations. The aim of this study was to investigate the associations of the three loci with HIV-1 infection and the progression of AIDS in the Han population of northern China. We recruited 78 HIV-1-seropositive individuals and 70 population-matched healthy controls from north China. PCR-RFLP was used to genotype these three polymorphisms in all samples. There were no significant differences in the frequencies of any allele between HIV-1-seropositive individuals and healthy controls. In cases, the associations between the three loci and CD4⁺ T-lymphocyte counts were not significant; however, there was a significant association between the genotype frequency of CCR2-64I and clinical category (P < 0.05). We have identified a potentially important role of CCR2-64I allele in AIDS progression in the northern Chinese population. Further studies are necessary to clarify the contribution of the CCR2-64I allele to the progression of AIDS for the relatively small sample size.