Abamectin resistance in Drosophila is related to increased expression of P-glycoprotein via the dEGFR and dAkt pathways

Many insects have evolved resistance to abamectin but the mechanisms involved in this resistance have not been well characterized. P-glycoprotein (P-gp), an ATP-dependent drug-efflux pump transmembrane protein, may be involved in abamectin resistance. We investigated the role of P-gp in abamectin (ABM) resistance in Drosophila using an ABM-resistant strain developed in the laboratory. A toxicity assay, Western blotting analysis and a vanadate-sensitive ATPase activity assay all demonstrated the existence of a direct relationship between P-gp expression and ABM resistance in these flies. Our observations indicate that P-gp levels in flies' heads were higher than in their thorax and abdomen, and that both P-gp levels and LC₅₀ values were higher in resistant than in susceptible and P-gp-deficient strains. In addition, P-gp levels in the blood–brain barrier (BBB) of resistant flies were higher than in susceptible and P-gp-deficient flies, which is further evidence that a high level of P-gp in the BBB is related to ABM resistance. Furthermore, we found greater expression of Drosophila EGFR (dEGFR) in the resistant strain than in the susceptible strain, and that the level of Drosophila Akt (dAkt) was much higher in resistant than in susceptible flies, whereas that in P-gp-deficient flies was very low. Compared to susceptible flies, P-gp levels in the resistant strain were markedly suppressed by the dEGFR and dAkt inhibitors lapatinib and wortmannin. These results suggest that the increased P-gp in resistant flies was regulated by the dEGFR and dAkt pathways and that increased expression of P-gp is an important component of ABM resistance in insects.     

microRNA-448 inhibits stemness maintenance and self-renewal of hepatocellular carcinoma stem cells through the MAGEA6-mediated AMPK signaling pathway

Hepatocellular carcinoma (HCC) occurs mainly in patients with chronic liver disease and cirrhosis. Increasing evidence has identified the involvement of microRNAs (miRNAs) acting as essential regulators in the progression of HCC. As predicted by microarray analysis, miR-448 might potentially affect HCC progression by regulating the melanoma-associated antigen (MAGEA). Therefore, the present investigation focused on exploring whether or not miR-448 and MAGEA6 were involved in the self-renewal and stemness maintenance of HCC stem cells. The interaction among miR-448, MAGEA6, and the AMPK signaling pathway was evaluated. It was noted that miR-448 targeted and downregulated MAGEA6, thus activating the AMP-activated protein kinase (AMPK) signaling pathway in HCC. Furthermore, for the purpose of exploring the functional relevance of MAGEA6 and miR-448 on the sphere formation, colony formation, and invasion and migration of HCC stem cells, the CD133⁺CD44 ⁺ HCC stem cells were sorted and treated with the mimic or inhibitor of miR-448, small interfering RNA (siRNA) against MAGEA6 or an AMPK activator AICAR. MAGEA6 silencing or miR-448 overexpression was demonstrated to inhibit the abilities of sphere formation, colony formation, cell migration, and invasion of HCC cells. Afterwards, a rescue experiment was conducted and revealed that MAGEA6 silencing reversed the effects of miR-448 inhibitor on stemness maintenance and self-renewal of HCC stem cells. Finally, after the in vivo experiment was carried out, miR-448 was observed to restrain the tumor formation and stemness in vivo. Altogether, miR-448 activates the AMPK signaling pathway by downregulating MAGEA6, thus inhibiting the stemness maintenance and self-renewal of HCC stem cells, which identifies miR-448 as a new therapeutic strategy for HCC.     

灰飞虱内生病毒HiPV外壳蛋白VP1多克隆抗体的制备及在病毒检测中的应用

【目的】前期发现水稻条纹病毒(rice stripe virus, RSV)可与介体灰飞虱Laodelphax striatellus体内的HiPV病毒(Himetobi P virus, HiPV)互作。本研究旨在制备HiPV外壳蛋白VP1的多克隆抗体,并评估其在HiPV病毒检测中的可用性,以为深入研究HiPV-RSV和HiPV-灰飞虱的互作机制提供技术支持。【方法】以RT-PCR方法从灰飞虱成虫体内扩增HiPV主要外壳蛋白基因VP1,然后将VP1基因亚克隆至原核表达载体pET-32a中,构建表达载体pET-VP1。将重组质粒转化大肠杆菌Escherichia coli BL21 (DE3),经IPTG诱导、Ni²⁺-NTA亲和层析纯化,获得重组蛋白,免疫新西兰大白兔,制备抗体。【结果】从灰飞虱体内克隆到774 bp的HiPV外壳蛋白基因VP1,经原核表达、纯化,获得分子量约47.5 kD的融合蛋白,免疫新西兰大白兔后获得VP1多克隆抗体。该抗体间接ELISA效价达1∶819 200,与HiPV外壳蛋白VP1有特异性反应,而与灰飞虱蛋白无交叉反应。利用该多克隆抗体建立了检测单头灰飞虱成虫体内HiPV的Western blot和免疫捕获RT-PCR方法,检测结果显示HiPV在携带和不携带RSV的灰飞虱高亲和性群体内均广泛存在。【结论】利用制备的HiPV的VP1多克隆抗体可特异性检测灰飞虱体内HiPV。本研究为HiPV病毒的快速检测以及HiPV-RSV互作、HiPV-灰飞虱互作研究提供了技术支持。     

Effect of tri-o-cresyl phosphate on cytoskeleton in human neuroblastoma SK-N-SH cell

Cytoskeletal components play an important role in maintaining cellular architecture and internal organization, with clear involvement of defining cell shape, in cell division and other cellular processes, such as neurite extension and maintenance. Alterations of cytoskeleton in human neuroblastoma SK-N-SH cells after exposure to different concentrations of tri-ocresyl phosphate (TOCP) for 12 hr were investigated. TOCP decreased the cell viability in a dose-dependent manner; the viability of SK-N-SH was reduced to approximately 50% of baseline after a 12-hour exposure to TOCP at high concentration (5 mM). Biochemical characterization by western blotting revealed that 1 and 5 mM concentrations of TOCP significantly inhibited the expression of neurofilament high molecular weight protein (NF-H), and that 5 mM TOCP inhibited expression of microtubule-associated protein 2c and tau protein, but not β-actin. Indirect immunofluorescence analysis revealed that higher concentrations of TOCP decreased the length of neuritis and changed the structure of microfilaments, which are associated with NF-H. In addition, activities of neuropathy target esterase and acetylcholinesterase were significantly reduced after exposure to 5 mM TOCP for 12 hr. Together, these results suggested that the loss of cytoskeletal components is the early event during the process of TOCP toxicity towards human neuroblastoma SK-N-SH cells.     

Identification and characterization of a chondroitin synthase from Avibacterium paragallinarum

Applied Microbiology and Biotechnology - Avibacterium paragallinarum is a Gram-negative bacterium that causes infectious coryza in chicken. It was reported that the capsule polysaccharides...     

玉米农田水热通量动态与能量闭合分析

 基于锦州农田生态系统野外观测站玉米农田涡度相关系统近2年的水热通量观测数据，分析了玉米农田水热通量的日际、年际变化特征及其能量 平衡状况。结果表明： 1）玉米农田水热通量日变化与年变化均呈单峰型二次曲线，峰值出现在12∶00～13∶00左右，与净辐 射的日变化、年 变化同步，潜热通量最大可达到655 w•m^－2（出现在2004年7月8日1 3∶00），显热通量最大值大约为369 w•m^－2（出现在2004年5月31日13∶ 00）。2）玉米农田水热通量强度与局地的环境条件密切相关：显热通量与大气压的年变化呈负相关，潜热通量与气温年变化呈正相关。水热通 量受降水的影响较大，对降水的反应较敏感。其中，潜热通量(LE)不仅与降水的强度有关，而且随着降水的季节分布的不同而出现不同的响应 ，即使同样量级的降水在夜间与白天对LE的影响也是不同的。3）玉米农田通量观测呈现能量不闭合现象，主要原因可能是未包含0～5 cm土壤 热储量与冠层热储量，造成大约15.5%的能量损失。     

A strategy for introducing an endangered plant <Emphasis Type="Italic">Mosla hangchowensis</Emphasis> to urban area based on nitrogen preference

To introduce endangered plants to urban green space for ex situ conservation successfully, it is important to better understand the optimal NO₃ ^?/NH₄ ⁺ ratios for profitable plant. Increasing nitrogen deposition altered the nitrate to ammonium ratio (NO₃ ^?/NH₄ ⁺) in soil. This change may strongly affect the fate of endangered plants, which often have little ability to adapt to environmental changes. In this study, we carried out a microcosm hydroponic experiment by growing Mosla hangchowensis (an endangered species) to test its preference to NO₃ ^?/NH₄ ⁺ ratios and used congeneric M. dianthera (a widespread species) for comparison. Results showed that M. hangchowensis preferred an equal NO₃ ^?/NH₄ ⁺ ratio to NO₃ ^? as an N source, with a higher biomass observed under NO₃ ^?/NH₄ ⁺ ratios of 50/50 and 75/25 than other treatments. However, M. dianthera preferred NO₃ ^? as N source, with a higher biomass under NO₃ ^?/NH₄ ⁺ ratios of 100/0 and 75/25 than other treatments. NH₄ ⁺ is the dominant form of N in atmospheric deposition in China and continued increasing in nitrogen deposition may be detrimental to M. hangchowensis, while only have minimal effects on M. dianthera. Urban regions are expanding, and the high environmental heterogeneity in urban areas can provide potential habitats for M. hangchowensis. Based on this study, we advise that the ex suit conservation of M. hangchowensis in urban green spaces needs to adjust the fertilization strategy according to the situation of nitrogen deposition to achieve the optimal NO₃ ^?/NH₄ ⁺ ratio.