Expression of recombinant and mosaic Cry1Ac receptors from Helicoverpa armigera and their influences on the cytotoxicity of activated Cry1Ac to Spodoptera litura Sl-HP cells

Bacillus thuringiensis (Bt) toxin receptors play important roles in the killing of pests, and investigation on characterization of the receptors is essential for utilization of Bt and management of insect resistance. Here, recombinant and mosaic receptors of Bt Cry1Ac toxin from Helicoverpa armigera were expressed in Spodoptera litura Sl-HP cells and their influences on cytotoxicity of activated Cry1Ac toxin were investigated. When H. armigera aminopeptidase N1 (APN1), alkaline phosphatase 2 (ALP2) and cadherin fused with or without GFP tag were, respectively, expressed in Sl-HP cells, live cell-immunofluorescence staining detection revealed that the quantity of the toxin binding to cadherin or cadherin-GFP was much more than that binding to ALP2 and APN1 or their fusion proteins with GFP, and only the cadherin- or cadherin-GFP-expressing cells showed aberrant cell morphology after the treatment of the toxin at low concentrations. ALP2 and APN1 fused with or without GFP tag did not significantly enhance the cadherin-mediated cytotoxicity of the toxin. The mosaic ALP-TBR-GFP-GPI was located on cell membrane, but did not bind to the toxin. The mosaic truncated cadherin-GFP-GPI was not located on cell membrane even if the signal peptide was sustained. The concentrations of the toxin resulting in swelling of 50 % cells for noncadherin-expressing Sl-HP cells and cadherin-expressing Hi5 cells were 5.08 and 9.50 µg/ml within 1 h, respectively. Taken together, our data have indicated that the binding affinity of ALP2 and APN1 to activated Cry1Ac toxin is much weaker than that of cadherin and both ALP2 and APN1 do not enhance the cytotoxicity of the toxin even though cadherin is co-expressed, and the mosaic receptor of ALP2 inserted with cadherin toxin binding domain does not mediate cytotoxicity of the toxin. In addition, the noncadherin-expressing Sl-HP cells are more susceptible to activated Cry1Ac than the cadherin-expressing Hi5 cells.     

鹤山南亚热带草坡生态系统的热量平衡

本文以广东鹤山丘陵综合试验站草坡为研究对象,分析了鹤山南亚热带草坡的能量平衡特征,主要结果如下：1．辐射平衡的各分量中,蒸发散耗热占净辐射的74.7％,其年变化幅度较大;湍流热通量占净辐射的21．2％,其月变化没有明显的规律,波动较大,土壤热通量与其它分量相比,其比重要小得多。2．虽然鹤山年幅射量和降雨量较多,但水热条件有几个月配合欠佳,使得草坡的净辐射能有90％以上用于地面以上植被和空气湍流交换所消耗,小气候条件较差,影响了这一生态系统的生产潜力的发挥。3．太阳辐射透过大气层以后,仅有4775．2MJm-2a-1左右的辐射能到达草坡,草坡反射了822．5MJm-2a-1,其净幅射收人为2915．6MJm-2a-1,净辐射收人中又有2077．8MJm-2a-1用于蒸散作用,618.1MJm-2a-1用于湍流热交换,87．5MJm-2a-1用于土壤热交换,植被贮热约38.8MJm-2a-1;用于净光合作用耗热93．4MJm-2a-1。     

海南岛河港海莲红树林凋落物动态的研究

 本文应用凋落物收集网对海南岛海莲红树群落进行连续4年(1984.1—1987.12)的凋落物动态研究。结果表明： (1)在保护良好的河港海滩上,海莲群落的年凋落物量高达1255g／m2,是热带地区凋落物量最大的群落类型之一;与世界上红树群落凋落物量相比处于较高水平。(2)4年平均凋落物各组分占总量的比例为叶64.32%、花10.63%,果21.34%、枝3.71%。 (3)月凋落物量(Y,g／m2)与月均气温(X,℃)的回归公式为 Y=5.009X–13.18(r=0.44**,df=46),相关极显著。 (4)不同年份凋落物的变化率R=1.33。     

<Emphasis Type="Italic">Lactobacillus plantarum</Emphasis> prevents the development of colitis in IL-10-deficient mouse by reducing the intestinal permeability

It is reported that defects exist in the small intestinal epithelial barrier of inflammatory bowel disease, which might be associated with increased intestinal permeability at a very early stage. Our study aims to investigate the role of Lactobacillus plantarum on the decrease of epithelial permeability and the further protective effects on the intestinal epithelial barrier using the IL-10-deficient mouse model. Our study showed that tight junction associated proteins were increased after the pre-treatment of L. plantarum by fluorescence staining, western blot, real-time PCR and transmission electron microscope. Oral gavage of milk containing L. plantarum was effective in decreasing small intestinal permeability using methods of Ussing chamber assay and sugar probe. Assay of pro-inflammatory cytokines IFN-γ, TNF-α, MPO, and colonic histology by ELISA showed protective effects of L. plantarum on the intestinal epithelial barrier. Therefore, L. plantarum may prevent the development of colitis in IL-10-deficient mice by blocking changes in the expression of TJ proteins, TJ structure and intestinal permeability.     

The Orphan Nuclear Receptor NR4A1 Protects Pancreatic β-Cells from Endoplasmic Reticulum (ER) Stress-mediated Apoptosis

The role of NR4A1 in apoptosis is controversial. Pancreatic β-cells often face endoplasmic reticulum (ER) stress under adverse conditions such as high free fatty acid (FFA) concentrations and sustained hyperglycemia. Severe ER stress results in β-cell apoptosis. The aim of this study was to analyze the role of NR4A1 in ER stress-mediated β-cell apoptosis and to characterize the related mechanisms. We confirmed that upon treatment with the ER stress inducers thapsigargin (TG) or palmitic acid (PA), the mRNA and protein levels of NR4A1 rapidly increased in both MIN6 cells and mouse islets. NR4A1 overexpression in MIN6 cells conferred resistance to cell loss induced by TG or PA, as assessed by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, and TUNEL assays indicated that NR4A1 overexpression also protected against ER stress-induced apoptosis. This conclusion was further confirmed by experiments exploiting siRNA to knockdown NR4A1 expression in MIN6 cells or exploiting NR4A1 knock-out mice. NR4A1 overexpression in MIN6 cells reduced C/EBP homologous protein (CHOP) expression and Caspase3 activation induced by TG or PA. NR4A1 overexpression in MIN6 cells or mouse islets resulted in Survivin up-regulation. A critical regulatory element was identified in Survivin promoter (−1872 bp to −1866 bp) with a putative NR4A1 binding site; ChIP assays demonstrated that NR4A1 physically associates with the Survivin promoter. In conclusion, NR4A1 protects pancreatic β-cells against ER stress-mediated apoptosis by up-regulating Survivin expression and down-regulating CHOP expression, which we termed as “positive and negative regulation.”     

Disruption of a receptor-mediated mechanism for intracellular sorting of proinsulin in familial hyperproinsulinemia

In familial hyperproinsulinemia, specific mutations in the proinsulin gene are linked with a profound increase in circulating plasma proinsulin levels. However, the molecular and cellular basis for this disease remains uncharacterized. Here we investigated how these mutations may disrupt the sorting signal required to target proinsulin to the secretory granules of the regulated secretory pathway, resulting in the unregulated release of proinsulin. Using a combination of molecular modeling and site-directed mutagenesis, we have identified structural molecular motifs in proinsulin that are necessary for correct sorting into secretory granules of endocrine cells. We show that membrane carboxypeptidase E (CPE), previously identified as a prohormone-sorting receptor, is essential for proinsulin sorting. This was demonstrated through short interfering RNA-mediated depletion of CPE and transfection with a dominant negative mutant of CPE in a beta-cell line. Mutant proinsulins found in familial hyperproinsulinemia failed to bind to CPE and were not sorted efficiently. These findings provide evidence that the elevation of plasma proinsulin levels found in patients with familial hyperproinsulinemia is caused by the disruption of CPE-mediated sorting of mutant proinsulins to the regulated secretory pathway.     

华西雨屏区香樟人工林土壤表层细根生物量和碳储量

2010年11月-2011年12月, 研究了华西雨屏区31年生香樟人工林土壤表层(0～30 cm)细根生物量及碳储量.结果表明: 香樟人工林土壤0～30 cm层细根总生物量(活根+死根)和碳储量的平均值分别为1592.29 kg·hm^-2和660.68 kg C·hm^-2,其中活细根贡献率分别为91.1%和91.8%.随着土壤深度的增加,香樟1～5级活细根和死细根的生物量及碳储量均显著减少;随着根序等级的升高,香樟活细根生物量及碳储量显著增加.香樟细根总生物量及碳储量均在秋季最高、冬季最低,死细根生物量及碳储量为冬季最高、夏季最低;1级根和2级根生物量及碳储量均在夏季最高、冬季最低,而3~5级根则为秋季最高、冬季最低.土壤养分和水分的空间异质性是导致细根生物量和碳储量变化的主要原因.     

Effects of Crosslinking Agent-DFDNB on Energy-transducing Reactions of Chloroplast Thylakoid Membrane Proteins

The effects of crosslinking agent-DFDNB (difluoro dinitro benzene) on functions of chloroplast thylakoid membrane proteins were investigated. DFDNB inhibited activities of PSP and membrane-bound ATPase in chloroplasts. It decreased proton uptake of light-inducted chloroplast thylakoids and the relative value of fluorescence quenching of 9-aminoacridine, and inhibited the rate of fast electrogenic phase of absorption change at 515 nm in chloroplasts. In addition, the isolated CF1-ATPase was crosslinked with DFDNB. The pattern of polymers of crosslinked CF1-ATPase was observed on SDS-PAGE.     

非人灵长类是否回避近亲繁殖?

近亲繁殖回避现象是人类学者、生物学者、心理学者和社会学者最关心的研究课题之一。针对灵长类的社会特点和近亲繁殖回避假说,本文引入了三个主要假设:假设1)近亲繁殖可能会导致个体繁殖适合度降低等损失;假设2)近亲繁殖有利有弊,但是弊端可能明显大于利益;假设3)如果弊大于利,那么动物应该进化形成回避近亲繁殖的机制,以减少该行为产生的损失。然后,本文综述分析了6科19个灵长类种群的现有繁殖数据,验证了灵长类的行为回避、迁移和繁殖抑制等机制,旨在强调灵长类回避近亲繁殖的必要性及其生物学背景,并为人类学、生物学和心理学的相关研究提供跨学科素材。