Background: Rheumatoid arthritis (RA) is a type of difficult-to-cure arthralgia with a worldwide prevalence. It severely affects people’s living standards. For a long time, bee venom has been used to treat RA and has shown good results. Melittin is the main active component of bee venom used for RA treatment, but the molecular mechanism of melittin in RA treatments remains unclear. Methods: Potential melittin and RA targets were obtained from relevant databases, and common targets of melittin and RA were screened. The STRING database was used to build the PPI network and screen the core targets after visualization. The core targets were enriched by Gene Ontology functional annotation and Kyoto Encyclopedia of Genes and Genomes pathway. Finally, the binding of melittin to target proteins was evaluated through simulated molecular docking, which verified the reliability of the prediction results of network pharmacology. Results: In total, 138 melittin targets and 5795 RA targets were obtained from relevant databases, and 90 common targets were obtained through intersection. Eighteen core targets, such as STAT3, AKT1, tumor necrosis factor, and JUN, were screened out. Enrichment analysis results suggested that melittin plays an anti-RA role mainly through tumor necrosis factor, interleukin-17, toll-like receptors, and advanced glycation end products–RAGE signaling pathways, and pathogenic bacterial infection. Molecular docking results suggested that melittin has good docking activity with core target proteins. Conclusion: RA treatment with melittin is the result of a multi-target and multi-pathway interaction. This study offers a theoretical basis and scientific evidence for further exploring melittin in RA therapy.

Glyphosate (GY) is the most widely used herbicide in agriculture worldwide. Lead is a common heavy metal in the natural environment. Honeybees, as pollinators, are exposed to these pollutants. So far, few reports have evaluated the toxic effects of GY mixed with heavy metals on honeybees (Apis cerana cerana). This study found that the acute toxicity of lead (LC50 = 1083 mg/L) is much greater than that of GY (LC50 = 4764 mg/L) at 96 h. The acute toxicities of the mixed substances were as follows: LC50 = 621 mg/L of lead and LC50 = 946 mg/L of GY. The combination of lead and GY was more toxic than either of the individual substances alone. Compared to the individual toxicity, combined treatment significantly affected the bees’ learning and cognitive abilities and changed the relative expression of genes related to immune defense and detoxification metabolism in A. c. cerana. The combination of lead and GY seriously affected the behavior and physiology of the studied honeybees. This study provides basic data for further research on the combined effects of GY and heavy metals on bee health. It also serves as a reference for effective colony protection.

Honey bees play a critical role as pollinators. However, their reproduction success and survival face severe threats due to the deterioration of their living environment. Notably, environmental conditions during their preimaginal stage inside brood cells can influence their immune capabilities and overall health after emergence. During the in-cell developmental stage, workers are in close contact with cocoons, which can become a source of stress due to accumulated metals. To investigate this potential threat, experiments were conducted to examine the impact of cocoons in brood cells used to rear different generations on the metal content and detoxification gene expression levels in Apis cerana cerana. Our findings indicated significant differences in the layers, weight, base thickness, and metal contents like Cr, Cd, Pb, Mn, Ni, and As of cocoons in multi-generation brood cells compared to single-generation brood cells. These increases led to significant elevations in metal levels and upregulations of the four CYP450 detoxification genes in both six-day-old larvae and newly emerged workers. In conclusion, this study highlights the negative impact of cocoons in multi-generation brood cells on bee health and provides evidence supporting the development of rational apiculture management strategies for ecosystem stability.

Background/Objectives: The larval stage plays a pivotal role in determining caste and sex in Apis mellifera. This study integrates RNA-seq, WGCNA, and alternative splicing analyses to explore gene expression differences among 2-day-old worker, drone, and queen larvae. Methods: RNA-seq was conducted on 2-day-old larvae from all three castes. Differential expression, WGCNA, and alternative splicing patterns were investigated. A deep learning TCN model was trained using WGCNA-derived modules and demonstrated high classification accuracy. Results: The TCN model highlighted a top-10 gene set, including PDHB, Fibroin3, and LOC724161. Significant caste- and sex-specific splicing events were detected in Tk, Csd, and Fem, with AF events being most prevalent. Splicing differences between sexes exceeded those observed among castes. Conclusions: The 2-day-old larval stage is crucial for both caste and sex differentiation in honeybees. This study identifies key genes and splicing events, offering new insights into the molecular mechanisms underlying caste formation and sex determination.

Background/Objectives: Early larval development is critical for caste and sex differentiation in honeybees. This study investigates molecular divergence in 4-day-old Apis mellifera larvae and introduces a customized deep learning model for hub-gene discovery. Methods: Genome-guided RNA-seq, DEGs, WGCNA, and splicing analyses were integrated. A hybrid convolution–attention model, ACmix-Swin, combined with WGAN-GP augmentation, was developed to classify larvae and prioritize caste-biased genes. Selected genes were validated by qPCR. Results: Significant caste- and sex-specific divergence was detected in cuticle formation, hormone metabolism, and reproductive signaling. ACmix-Swin achieved the highest accuracy among baseline models and consistently identified key regulators, including Vg, LOC725841, LOC412768, and LOC100576841. qPCR confirmed RNA-seq trends. Conclusions: Caste- and sex-specific transcriptional programs are established early in larval development. The ACmix-Swin framework provides an effective strategy for high-dimensional transcriptome interpretation and robust hub-gene identification.

The size of comb cells is a key factor influencing the body size of honey bee workers. Comb cells and the body size of Chinese honey bee workers are smaller than those of Italian honey bee workers. To increase the size of Chinese honey bee workers, this study used newly built combs from Chinese honey bee colonies (control group) and Italian honey bee colonies (treatment group). These combs were provided to Chinese honey bee colonies for queens to lay fertilized eggs with the aim of rearing larger workers. Workers emerging from the control and treatment combs were designated as control and treatment workers, respectively. We compared 13 external morphological traits, including right forewing length and width; linear length of veins a, b, c, and d; proboscis length; right hind femur length; tibia length; metatarsal length and width; and the longitudinal diameters of the third and fourth tergites between the two groups. The results identified six types of cell contents in the combs, excluding empty cells: capped honey (most abundant), followed by capped brood, uncapped honey, and smaller amounts of pollen, larvae, and eggs. Additionally, the average body weights of 6-day-old worker bee larvae, white-eyed pupae, adult worker bees, and honey stomachs containing sucrose solution were significantly higher in the treatment group than in the control group. Except for proboscis length, the average size of 12 out of 13 traits in the treatment group was significantly larger than in the control group, indicating that increasing cell size can produce larger Chinese honey bee workers. In the control group, only the left forewing was significantly shorter than the right, with no significant side-to-side differences in the other morphological traits. In the treatment group, the left forewing width, hind leg femur length, tibia length, and hind leg metatarsus width were significantly smaller than their right-side counterparts, while the other six traits showed no significant side-to-side differences. These findings provide a basis for using Italian honey bee combs to rear larger workers in Chinese honey bee colonies, offering beekeepers a strategy to enhance foraging capacity and improve honey production.