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Lipids are main energy source for insects reproduction, which are becoming emerging target for pest management. Azadirachtin (AZA) is a multi-targeted and promising botanical insecticide, but its reproduction toxicity mechanism related to lipids metabolism is poorly understood. Here, we applied lipidomic and transcriptomic to provide a comprehensive resource for describing the effect of AZA on lipids remodeling in ovary of Spodoptera litura. The results showed that AZA exposure obviously altered the contents of 130 lipids subclasses (76 upregulated and 54 downregulated). In detail, AZA exposure changed the length and saturation degrees of fatty acyl chain of most glycerolipid, phospholipid and sphingolipid as well as the expression of genes related to biosynthesis of unsaturated fatty acids and fatty acids elongation. Besides, following the abnormal lipids metabolism, western blot analysis suggested that AZA induce insulin resistance-like phenotypes by inhibiting insulin receptor substrates (IRS) /PI3K/AKT pathway, which might be responsible for the ovary abnormalities of S. litura. Collectively, our study provided insights into the lipids metabolism event in S. litura underlying AZA exposure, these key metabolites and genes identified in this study would also provide important reference for pest control in future.

Summary: Lysosome motility is critical for the cellular function. However, Rab7-related transport elements showed genetic differences between vertebrates and invertebrates, making the mechanism of lysosomal motility mysterious. We suggested that Rab7 interacted with RILP as a feature of highly evolved organisms since they could interact with each other in Spodoptera frugiperda but not in Drosophila melanogaster. The N-terminus of Sf-RILP was identified to be necessary for their interaction, and Glu61 was supposed to be the key point for the stability of the interaction. A GC-rich domain on the C-terminal parts of Sf-RILP hampered the expression of Sf-RILP and its interaction with Sf-Rab7. Although the corresponding vital amino acids in the mammalian model at the C-terminus of Sf-RILP turned to be neutral, the C-terminus would also help with the homologous interactions between RILP fragments in insects. The significantly different interactions in invertebrates shed light on the biodiversity and complexity of lysosomal motility.

Azadirachtin is one of the most successful botanical pesticides in agricultural pest control. To build a repertoire of proteins and pathways in response to azadirachtin exposure during ovarian development, iTRAQ-based comparative proteomic was conducted. 1423 and 1686 proteins were identified as differentially accumulated proteins (DAPs) by comparing the protein abundance in adult ovary with that in pupal ovary under normal and azadirachtin exposure condition, respectively. Bioinformatics analysis indicated that pupae-to-adult transition requires proteins related to proteasome and branched chain amino acids (BCAAs) degradation for ovary development. Azadirachtin exposure strongly affected glycosylation-related pathway. And proteins related to vitamin B6 synthesis were necessary for ovary development under normal and AZA-exposure condition. RNAi assays confirmed the essential roles of DAPs related to glycosylation and vitamin B6 synthesis in moth growth and ovary development. The results enhance our understanding of the molecular regulatory network for ovary development and provide valuable resources for using AZA-responsive proteins to develop novel bio-rational insecticides.

Chemical pesticides and adjuvants have caused many negative effects. Botanical compounds provide solutions for the development of environment friendly pesticides and the management of increasing pest resistance. Curcumin, a natural polyphenol, showed synergistic effects on avermectin upon the destructive agricultural pest, Spodoptera litura. However, the botanical synergist and its relevant mechanisms remain unclear. In the article, curcumin significantly enhanced the growth inhibition and midgut structural damage of avermectin on the larvae of S. litura, and the synergistic effects were confirmed with pot experiments. There were only a few influences on the gene expression of avermectin targets, while apoptotic and autophagic related genes and proteins were accumulated in the avermectin/curcumin mixed regent (0.013/0.0013 μg/mL) treated group. Moreover, the potential mechanism was explored with an in vitro model, insect Spodoptera frugiperda Sf9 cell line. Morphology observation featured the damage on cells and Hoechst33258 staining revealed the fragments of DNA after treating with the avermectin/curcumin mixed regent (10/1 μg/mL). Dansylcadaverine and LysoTracker staining, as well as the gene expressions, supposed that curcumin exhibited autophagy inducing effects and the mixed regent possessed a higher ability to induce apoptosis and autophagy. All these results suggested that the synergistic effects of curcumin on the pest management of avermectin potentially mainly derived from the enhancement of programed cell death. It provides new sights for the application of natural compounds in integrated pest management and enriches examples of synergistic mechanisms.

Objective: To measure vascular endothelial function (VED) in obese/overweight people grouped according to the constitutional theory of traditional Chinese medicine (TCM). We wished to predict the risk of VED and cardiovascular diseases in obese/overweight populations. Methods: This was a cross-sectional study. Eighty-six obese/overweight volunteers from Beijing Chao-Yang Hospital were enrolled and divided into four groups based on body-constitution type: phlegm dampness (PD); phlegm dampness with blood stasis (PDBS); qi-deficiency (QD); balance (BA). Height, weight, waist circumference and hip circumference were measured, and percent body fat and reactive hyperemia index (RHI) calculated. Levels of vascular endothelial growth factor (VEGF), von Willebrand factor (vWF) and plasma endothelial-1 (ET-1) were detected by immunohistochemistry. Results: The RHI and vWF levels in the PDBS group were significantly lower than those in the BA group and QD group (P .05). These two indicators displayed no significant differences among BA, QD, and PD groups (P > .05). There was no significant difference among the four groups in level of ET-1 or VEGF (P > .05). Conclusion: Of the four indicators selected, the RHI and vWF displayed significant differences between the PDBS group and BA group, and between the PDBS group and QD group. Hence, in an obese/overweight population with different constitutional types, compared with BA and QD groups, a relatively higher risk of suffering from VED was detected in the PDBS group. Meanwhile, this study provides the experimental basis for confirming the feasibility of constitutional classification in obese/overweight population. Keywords: Obese/overweight, Constitution classification, Vascular endothelial dysfunction, Reactive hyperemia index

Objective: To explore the possible correlation between traditional Chinese medicine (TCM) constitution and facial features in color images and to improve the accuracy of automated constitution classification. Methods: Color images were taken of 5150 individuals of different professions. Automated face detection and key point positioning were performed on the collected images, which were then transformed into a standard size. The relationship between facial features and TCM constitution based on the red, green, blue (RGB) pixel and the local binary pattern (LBP) texture features was explored. Results: The overall accuracy rate and robustness of TCM constitution classification based on RGB features were low. Classification results of the phlegm-dampness, damp-heat, blood stasis, and balance constitutions achieved high accuracy rates. Classification accuracy rate using the LBP texture feature was higher than that of the RGB feature, with the best accuracy observed for the balance constitution. Conclusion: Application of computer image acquisition and processing of facial features may serve as an adjunct to the TCM diagnostic method of inspection.