Wasabi leaf extracts attenuate adipocyte hypertrophy through PPARγ and AMPK

Hypertrophy of adipocytes in obese adipose tissues causes metabolic abnormality by adipocytokine dysregulation, which promotes type 2 diabetes mellitus, hypertension, and dyslipidemia. We investigated the effects of wasabi (Wasabia japonica Matsum) leaf extracts on metabolic abnormalities in SHRSP.Z-Leprfa/IzmDmcr rats (SHRSP/ZF), which are a model of metabolic syndrome. Male SHRSP/ZF rats aged 7 weeks were divided into two groups: control and wasabi leaf extract (WLE) groups, which received water or oral treatment with 4 g/kg/day WLE for 6 weeks. WLE improved the body weight gain and high blood pressure in SHRSP/ZF rats, and the plasma triglyceride levels were significantly lower in the WLE group. Adipocyte hypertrophy was markedly prevented in adipose tissue. The expression of PPARγ and subsequent downstream genes was suppressed in the WLE group adipose tissues. Our data suggest that WLE inhibits adipose hypertrophy by suppressing PPARγ expression in adipose tissue and stimulating the AMPK activity by increased adiponectin.     

Fluorescent labeling for clonal selection of Marc 145 cells secreting high levels of recombinant protein PBD-1

Despite the powerful impact gene expression markers like the green fluorescent protein (GFP) or enhanced GFP (EGFP) exert on linking the expression of recombinant protein for selection of high producers in recent years, there is still a strong incentive to develop more economical and efficient methods for isolating mammalian cell clones secreting high levels of recombinant proteins. Here we present a new method based on the co-expression of EGFP that allows clonal selection in standard 96-well cell culture plates. The genes encoding the EGFP protein and the related protein are linked by an internal ribosome entry site and thus are transcribed into the same mRNA in an independent translation process. Since both proteins arise from a common mRNA, the EGFP expression level correlates with the expression level of the therapeutic protein in each clone. By expressing recombinant porcine β-defensin 1 in Marc 145 cells, we demonstrate the robustness and performance of this technique. The method can be served as an alternative to identify high-producer clones with various cell sorting methods.     

Isolation and Identification of Pathogens Causing Marigold (Tagetes erecta L.) Black Spot in Beijing,China

Black spot leads to great marigold losses worldwide. The disease is characterized by black spots on leaves and stems in its early stages, and the whole plant has black rot at the advanced stage. In this report, 6 of 217 Alternaria strains isolated from lesions of marigold plants in Beijing were randomly selected. The morphological characteristics and a pathogenic tree based on two protein‐coding genes (gpd and alt a 1) indicated that Alternaria tagetica is the causal agent of marigold black spot in Beijing. All six Alternaria strains could successfully re‐infect marigold, but they could not infect carrot or zinnia by either spore spray in a greenhouse or planting experiments in the epidemic area. This is the first report of the A. tagetica pathogen being isolated from marigold in Beijing.     

Characterization of cell structural change,growth, lipid accumulation,and pigment profile of a novel oleaginous microalga, <Emphasis Type="Italic">Vischeria stellata</Emphasis> (Eustigmatophyceae), cultured with different initial nitrate supplies

The appropriate microalgal species and the optimal nitrogen supply in culture medium are vital factors in maximizing biomass and metabolite productivities in microalgae. Vischeria stellata is an edaphic unicellular eustigmatophycean microalga. Cytological and ultrastructural characteristics and the effects of different initial nitrate-nitrogen concentrations on growth, lipid accumulation, fatty acid profile, and pigment composition were investigated in the present study. The cell structures of V. stellata changed with the degree of nutrient utilization and growth phase. The initial nitrate concentration for the optimal growth of V. stellata ranged from 6.0 to 9.0 mM. The maximum total lipid (TLs), neutral lipid (NLs), and total fatty acid (TFAs) contents were 55.9, 51.9, and 44.7 % of dry biomass, respectively. The highest volumetric productivity of TLs, NLs, and TFAs reached 0.28, 0.25, and 0.21 g L^?1 day^?1, respectively. V. stellata had a suitable fatty acid profile for biodiesel production, as well as containing eicosapentaenoic acid (EPA) for nutraceutical applications. In addition, the content β-carotene, increased gradually as culture time was prolonged, resulting in its exclusive production at the end of cultivation. V. stellata is a promising microalgal strain for the production of biofuels and bioproducts.     

A Cocktail ELISA Semi‐nested RT‐PCR Assay to Detect Bean pod mottle virus in Soya bean Seeds

Bean pod mottle virus (BPMV) has been identified as an important pathogen for plant quarantine in China because large quantities of soya bean seeds (approximately 7 × 10⁷ tons) are imported annually. To develop a practical detection programme for BPMV, a cocktail enzyme‐linked immunosorbent assay (ELISA) nested RT‐PCR using a combination of serological and molecular methods was designed for soya bean seeds. The single‐vessel detection assay was performed in a 96‐well ELISA plate, which served as a carrier for the subsequent nested RT‐PCR assay. Assay specificity was demonstrated by the production of the expected 330‐ and 296‐bp bands using the external and internal primers, respectively. This method was 10⁴‐fold more sensitive than immunocapture‐RT‐PCR (IC‐RT‐PCR). In particular, it is important to note that this assay resulted in successful micro‐extraction from soya bean seeds and combined the advantages of each individual technique. The cocktail ELISA nested RT‐PCR is a specific, sensitive, rapid and economical procedure to rapidly identify and characterize BPMV and could be suitable for both primary‐level platforms and laboratories.     

四川米仓山自然保护区台湾水青冈群落学特征沿海拔梯度的变化

为了解不同生境中台湾水青冈(Fagus hayatae)的群落特征,对四川米仓山国家级自然保护区不同海拔的台湾水青冈群落的物种组成、区系特征、生活型谱、重要值和物种多样性等进行了研究。结果表明,台湾水青冈群落中的植物种类随海拔存在差异。群落中植物区系均以北温带分布类型为主,且其分布比例与海拔呈正相关关系。不同海拔群落的物种生活型谱主要以高位芽植物为主,其它生活型较少。群落乔木层中台湾水青冈的重要值随海拔上升不断增大。中海拔群落中的物种多样性指数均低于高海拔和低海拔,具有物种种类少、多样性低的特点。因此,不同海拔段上的台湾水青冈群落学特征有明显的差异。     

鄱阳湖浮游甲壳动物群落结构特征

鄱阳湖是中国第一大淡水湖,具有"丰水为湖,枯水为河"的独特特点。为探讨鄱阳湖浮游甲壳动物群落结构及其时空分布的特征,于2009年全年采集其不同季节、不同水位期的浮游甲壳动物样品进行定量分析。结果显示,鄱阳湖浮游甲壳动物群落结构总体与河流浮游甲壳群落具有相似性。无节幼体、象鼻溞、剑水蚤等河流优势类群在鄱阳湖浮游甲壳动物中占优势;而哲水蚤和溞属仅在低水位季节占优势,枝角类丰度仅在高温、高水位、流速缓的季节高过桡足类。丰水期浮游甲壳动物平均丰度和生物量远远高于枯水期,可达枯水期的50倍,差异极其显著(P0.01)。温度和水位变化引起的环境因子改变是导致鄱阳湖浮游甲壳动物发生季节演替的主要原因;而营养盐对浮游甲壳动物的影响并不显著。空间上浮游甲壳动物群落构成明显不同,年均丰度最高和最低的点均出现在河口地区。因此:对于换水周期短,水交换速率快的水体,应该充分考虑水文条件对生物的影响。     

竹产业生态系统结构及演化规律——以贵州省赤水市为例

以"中国竹子之乡"命名的贵州省赤水市为例,研究了竹产业生态系统结构及演化规律。结果表明:(1)竹子各构件都有开发价值,包括生态服务、食药、竹材加工和文化产品等5方面,产业系统呈3级链环模式,第二、第三产业链环由近200家加工企业及6大系列近300个竹加工品种组成,包括一般加工和深加工产品,第一产业链环主要由占全市森林面积47.3%的竹林资源构成。第一产业链环是环境保障与原材料的基础,第二和第三产业链环是产业化能力,三者互相依存。(2)竹产业生态系统由环境资源、原生产业、外生产业、共生产业和产业创新5个子系统组成,子系统间相互作用、重要性不同,出现原生产业子系统为主(Ⅰ)、外生-原生产业子系统为主(Ⅱ)、原生-外生产业子系统为主(Ⅲ)和外生产业子系统为主(Ⅳ)4种状态,赤水市竹产业生态系统处于Ⅱ→Ⅲ级状态。竹外生产业子系统产值占全市GDP的40%以上、呈逐年上升的趋势,2014年竹产业生态系统中原生、外生和共生产业子系统产值分别为0.95×10~9、2.85×10~9元和2.30×10~9元,总产值6.10×10~9元。(3)依据各子系统间相互作用及生产要素转化过程,赤水市竹产业生态系统演化过程划分为4个阶段。原始发展阶段(约1950年以前)处于Ⅰ级状态,手工利用阶段(1951—1980年)整体处于Ⅰ级、毛竹林资源及加工利用部分有向Ⅱ级转化的趋势,工业利用阶段(1981—2000年)处于Ⅱ级状态,系统利用阶段(2001年至今)整体处于Ⅱ级、杂竹林资源及加工利用部分有向Ⅲ级转化的趋势。现时竹产业发展需提高原生产业子系统规模与生产力,依靠产业创新子系统引领竹产业生态系统走创新驱动之路。     

野鸦椿种子内源抑制物活性初探

以野鸦椿(Euscaphis japonica)种壳和胚为材料,甲醇浸提得到种壳浸提液和胚浸提液,配制浸提液浓度梯度为原浸提液浓度的10%、20%、30%、40%,研究不同浓度的种壳和胚浸提液对白菜、小麦、绿豆种子底物酶活性、发芽率、幼苗根长和苗高的影响,萃取和分离种壳和胚甲醇浸提液中的内源抑制物质,探讨野鸦椿种子内源抑制物质的活性与成分。结果表明:随着种壳和胚浸提液浓度的增加,白菜种子酸性磷酸酶活性和发芽率均显著降低(P0.05),表现为抑制作用递增,种壳的抑制作用小于胚,而幼苗的根长和苗高则表现为低促高抑,在10%浸提液处理下根长和苗高达最大值,种壳的促进效果弱于胚;小麦种子淀粉酶活性及幼苗的根长和苗高递减(P0.05),表现为抑制作用递增,而发芽率则在浓度≤20%时差异不明显(P0.05),浓度为30%时显著降低(P0.05),40%时发芽率为0,种壳的抑制作用大于胚;绿豆种子蛋白酶活性、发芽率、幼苗根长和苗高均在浸提液浓度≥20%时显著下降(P0.05),且种壳的作用效果小于胚。种子内源抑制物萃取及分离表明,外壳中含酚酸类较胚多,含碱类较胚少。综上认为,野鸦椿种壳和胚中均含有较高活性的内源抑制物,但性质、成分及含量存在差异,外壳内源抑制物主要作用对象为淀粉类物质,胚乳内源抑制物主要作用于油脂类和蛋白类物质。     

白花除虫菊化学成分及其生物活性的研究

该研究应用柱层析法、薄层层析法和波谱法对白花除虫菊(Pyrethrum cinerariifolium)全株甲醇提取物进行分离纯化、结构鉴定,并分别采用 MTT 法、生物活性测定法和抑菌圈法测定所得化合物抗肿瘤、杀线虫和抑菌活性。结果表明：经波谱法鉴定化合物结构分别为 tulirinol (1),sesamin (2),β-cyclopyrethrosin (3)和3,5-dihydroxy-2-(4-hydroxy-3-methoxyphenyl)-7,8-dimethoxy-4H-1-benzopyran-4-one (4)。抗肿瘤活性显示化合物3对白血病细胞株 HL-60、肝癌细胞株 SMMC-7721、肺癌细胞株 A-549、乳腺癌细胞株 MCF-7和结肠癌细胞株SW480均表现出显著的抑制活性,其 IC50分别为3.800、2.890、2.930、4.600和5.160μmol?L-1;化合物1活性比3稍弱,其 IC50分别为5.020、10.760、12.310、12.310和12.250μmol?L-1;其中化合物3对各肿瘤细胞株 IC50值均小于顺铂。抗菌活性表明化合物3对大肠杆菌、蜡样芽孢杆菌、枯草芽孢杆菌和金黄色葡萄球菌均表现出明显的抑菌活性,且随着浓度增加活性逐渐增强,而化合物1和化合物2仅对部分菌株表现出微弱抑菌活性;杀线虫活性显示化合物3对秀丽隐杆线虫和全齿复活线虫均表现出显著的毒杀活性,且随着时间、浓度增加活性逐渐增强;而在同一时间点对秀丽隐杆线虫的毒杀活性强于全齿复活线虫。从白花除虫菊中分离所得4个化合物均为首次从该植株中发现,且首次报道了化合物3杀线虫和抑菌活性,值得进一步开发应用。