Bacillus thuringiensis Cry1Ac toxin binds to midgut proteins, as cadherin (CAD) and ABCC2 transporter, to form pores leading to larval death. In cell lines, co-expression of CAD and ABCC2 enhance Cry1Ac toxicity significantly, but the mechanism remains elusive. Here, we show that the expression of Helicoverpa armigera CAD (HaCAD-GFP) in Hi5 cells induces susceptibility to Cry1Ac and enhanced Cry1Ac toxicity when co-expressed with H. armigera ABCC2 (HaABCC2-GFP), since Cry1Ac toxicity increased 735-fold compared to Hi5 cells expressing HaCAD-GFP alone or 28-fold compared to HaABCC2-GFP alone. In contrast, the expression of the Spodoptera litura CAD (SlCAD-GFP) in Hi5 cells did not induce susceptibility to Cry1Ac nor it potentiated Cry1Ac toxicity with HaABCC2-GFP. To identify the CAD regions involved in the enhancement of Cry1Ac toxicity with ABCC2, the different CAD domains were replaced between SlCAD-GFP and HaCad-GFP proteins, and cytotoxicity assays were performed in Hi5 cells in the absence or presence of HaABCC2-GFP. The HaCAD toxin-binding region (TB), specifically the CAD repeat-11, was necessary to enhance Cry1Ac toxicity with ABCC2. We propose that CAD TB is involved in recruiting Cry1Ac to localize it in a good position for its interaction with the ABCC2, resulting in efficient toxin membrane insertion enhancing Cry1Ac toxicity.

Arma chinensis is a natural enemy that preys on various species and can suppress agricultural and forest pests in the orders Lepidoptera and Coleoptera. Here, we aimed to determine the genome of A. chinensis assembled at the chromosome-level using PacBio and Hi-C technologies. The assembled genome was 986 Mb, with a contig N50 of 2.40 Mb, scaffold N50 of 134.98 Mb, and BUSCO completeness of 96.10%. Hi-C data aided in anchoring the assembly onto seven chromosomes. A sequence of ~ 496.2 Mb was annotated as a repeat element, constituting 51.15% of the genome. We functionally annotated 84.79% of 20,853 predicted protein-encoding genes. This high-quality A. chinensis genome provides a novel genomic resource for future research on Pentatomidae insects.

Objectives: Nasopharyngeal carcinoma (NPC) is a type of nasopharyngeal disease with high metastasis and invasion properties. Tumor-associated alternative activated (M2) macrophages are evidenced to connect with NPC. Based on this, this study purposes to explore the mechanism and participation of microRNA-18a (miR-18a) from M2 macrophages in NPC. Methods: Peripheral blood mononuclear cells were differentiated to macrophages and macrophages were polarized to M2 type by interleukin-4. SUNE-1 and CNE2 cells were transfected with restored or depleted miR-18a or transforming growth factor-beta III receptor (TGFBR3) to explore their roles in NPC progression with the involvement of the TGF-β signaling pathway. Next, SUNE-1 and CNE2 cells were co-cultured with M2 macrophages that had been treated with restored or depleted miR-18a or TGFBR3 to comprehend their combined roles in NPC with the involvement of the TGF-β signaling pathway. Results: MiR-18a was highly expressed and TGFBR3 was lowly expressed in NPC cells. MiR-18a restoration, TGFBR3 knockdown or co-culture with miR-18a mimics, or si-TGFBR3-transfected M2 macrophages promoted SUNE-1 cell progression, tumor growth in mice, decreased p-Smad1/t-Smad1, and elevated p-Smad3/t-Smad3. miR-18a downregulation, TGFBR3 overexpression, or co-culture with miR-18a inhibitors or OE-TGFBR3-transfected M2 macrophages depressed CNE2 cell progression, tumor growth in mice, increased p-Smad1/t-Smad1, and decreased p-Smad3/t-Smad3. Conclusion: Our study elucidates that miR-18a from M2 macrophages results in promoted NPC cell progression and tumor growth in nude mice via TGFBR3 repression, along with the Smad1 inactivation and Smad3 activation.

Natural enemies of arthropods contribute considerably to agriculture by suppressing pests, particularly when combined with chemical control. Studies show that insect recovery after insecticide application is rare. Here, we discovered the recovery of the predatory bug Arma chinensis from knockdown following the application of β-cypermethrin but not five other insecticides. A. chinensis individuals were more tolerant to β-cypermethrin than lepidopteran and coleopteran larvae, which did not recover from knockdown. We assessed A. chinensis recovery by monitoring their respiration and tracking locomotion through the entire process. We identified and verified the trans-regulation of detoxifying genes, including those encoding cytochrome P450s and α/β-hydrolase, which confer recovery from β-cypermethrin exposure in A. chinensis, by mitogen-activated protein kinase (MAPK) and cAMP response element binding protein (CREB). Furthermore, we discovered a novel mechanism, the neurotransmitter clearance, in vivo during the recovery process, by which the insect initiated the removal of excessive dopamine with a degrading enzyme ebony. Overall, these results provide mechanistic insights into the detoxification and neurotransmitter clearance that jointly drive insect recovery from insecticide exposure.