Background: Attractive Toxic Sugar Baits (ATSBs) are an innovative vector control strategy based on the “attract-and-kill” principle. The core of ATSBs lies in the preparation of attractive and toxic baits through the mixing and proportioning of luring and active ingredients. Although previous studies have investigated the effects of ATSBs on mosquitoes, significant challenges remain for broader field application. Methods: This study evaluated five fruit juices as ATSBs for mosquitoes, focusing on feeding preferences. Preservative concentrations were assessed by measuring antimicrobial activity over time. Two commercial traps were tested for mosquito entry rates. The optimal insecticide species and concentration were determined based on mortality rates. An optimized ATSBs system was developed and tested under a semi-field cage. Statistical analysis was performed using GraphPad Prism. Results: Within 24 h, apple juice-based ATSBs had the highest attractant index for Culex quinquefasciatus and Anopheles sinensis, while a pear juice-based ATSB was most effective for Aedes albopictus. A 0.1% preservative concentration best maintained juice stability. The LC50 values of dinotefuran-based ATSBs for Cx. quinquefasciatus, Ae. albopictus, and An. sinensis were 1.18 × 10⁻³, 4.06 × 10⁻⁴, and 5.20 × 10⁻⁵ g/L, respectively. The Spodoptera frugiperda trap outperformed the Drosophilidae trap. Simulated semi-field cage tests showed 48 h mortality rates of 86.00% for Cx. quinquefasciatus and 95.67% for Ae. albopictus. Conclusion: This study optimized an ATSB system by screening various fruit juices, preservative concentrations, insecticides, and trap devices. The system’s efficacy in mosquito control was evaluated under a semi-field cage. These findings provide a strong foundation for the future application and refinement of ATSB-based mosquito control strategies.

Ricin toxin binding subunit B (RTB) is a galactose-binding lectin protein derived from the beans of the castor oil plant (Ricinus communis). Our previous studies have reported a direct immunomodulatory effect of recombinant RTB, which stimulates RAW264.7 cells to produce cytokines including TNF-α. However, the role of RTB in innate immune response and its specific mechanism have not been reported in detail. In this work, the results showed that RTB treatment of macrophages significantly increased TLR4 protein levels. RTB also activated TLR4 downstream events, including MyD88, IRAK, and TRAF6, resulting in macrophage activation and TNF-α production. This process is reflected in the increase of IκB phosphorylation. TLR4 knockdown macrophages treated with RTB exhibited greatly reduced IκB phosphorylation and TNF-α secretion. Moreover, treatment with MyD88 inhibitor also suppressed TNF-α production. The docking of RT and TLR4 was simulated by computer, and the contact residues were concentrated on RTB. Our results suggest that recombinant RTB can activate mouse macrophages to secrete TNF-α through activation of NF-κB via the TLR4 signaling pathways.

Increasing studies have concentrated on investigating circular RNAs (circRNAs) as pivotal regulators in the progression of numerous diseases and biological processes and abundant evidence shows that circRNAs are participated in the regulation of innate immune responses. Several studies showed that Ricin Toxin (RT) could induce inflammatory injury. There was no research on the particular functions and underlying mechanisms of circRNAs in RT-induced inflammation. In this study, RNA sequencing performed on RT-treated and normal RAW264.7 macrophage cells was used to investigated the differentially expressed circRNAs. Based on the dataset, the expression of circEpc1 (mmu_circ_0,000,842) was identified higher in RT-treated cells. Moreover, gain-and-loss function assays showed that circEpc1 function as a promoter in RT-induced inflammation in vivo and in vitro. Mechanistically, circEpc1 acted as a miR-5114 sponge to relieve the suppressive effect of miR-5114 on its target NOD2 and thereby activating NF-κB and MAPK signaling pathways. Our results illuminated a link between RT-induced inflammation and the circEpc1 regulatory loop and provided novel insight into the functions of circRNA in innate immune, which may emerge as a potential target in immunotherapy to control the RT-induced inflammatory injury.

Ricin toxin (RT), extracted from castor bean, is a type II ribosome-inactivating protein (RIP). As a biothreat agent, there is no effective antidote for RT to date, but recent advances in antibody research may have meet this need. This study aimed to produce human single-chain antibody variable fragments (HuscFvs) that bind to and interfere with RT activity for further clinical use. The purified RT component was used in phage biopanning to select ricin-bound HuscFv-displayed phage clones from synthetic (Tomlinson I + J) phage display libraries. The selected HuscFv, named JE11, showed an almost 100% protective effect to HeLa cells by mixing with RT at a 1:1000 ratio in vitro. The antibody showed an 83% protection against injection of 2 × LD₅₀ RT dose mice within 72 h. This research has therapeutic potential for the diagnosis and treatment of RT poisoning.