This study aims to reveal the interannual and seasonal variations in functional components in Polygonatum cyrtonema Hua. rhizomes and evaluate whether the variations significantly affect the quality of rhizomes as a traditional Chinese herbal medicine. The interannual and seasonal variations in total flavonoid content and total saponin content were analyzed. The global dynamic variation in secondary metabolites in the rhizomes during a five-year growth period and in two traditional harvesting seasons were investigated based on metabolomics method. Results clearly showed that the functional components in P. cyrtonema rhizomes exhibited a significant increase in accumulation during the one- to four-year growth period and a significant decrease in accumulation during the four- to five-year growth period. The most active accumulation occurred during the three- to four-year growth period. Drastic variations in functional components occurred from spring to autumn. The significant interannual variation and drastic seasonal variation were strongly associated with the enrichment in some pathways related to the biosynthesis of secondary metabolites and the metabolisms of amino acids. The interannual and seasonal variations in functional components significantly affected the quality of P. cyrtonema rhizomes. The four-year-old rhizomes had the most superior quality due to their higher content of functional components and much more newly formed components. The rhizomes harvested in spring or autumn had unequal quality because of their significant differences in composition and content of functional components. Specifically, the rhizomes from spring contained more flavonoids, alkaloids, and phenolic acids, while those from autumn comprised more steroids. In conclusion, this study reveals that the interannual and seasonal variations in functional components can significantly affect the quality of P. cyrtonema rhizomes as a traditional Chinese herbal medicine. This study provides foundational insights and theoretical guidance for determining an optimal cultivation period to obtain medicinal rhizomes with superior quality. It also offers a strategy for harvesting medicinal rhizomes in two different seasons to achieve unequal quality.

Insects have a robust capacity to produce offspring for propagation, and the reproductive events of female insects have been achieved at the molecular and physiological levels via regulatory gene pathways. However, the roles of MicroRNAs (miRNAs) in the reproductive development of the brown planthopper (BPH), Nilaparvata lugens, remain largely unexplored. To understand the roles of miRNAs in reproductive development, miRNAs were identified by Solexa sequencing in short-winged (SW) female adults of BPH. Small RNA libraries derived from three developmental phases (1 day, 3 days, and 5 days after emergence) were constructed and sequenced. We identified 905 miRNAs, including 263 known and 642 novel miRNAs. Among them, a total of 43 miRNAs were differentially expressed in the three developmental phases, and 14,568 putative targets for 43 differentially expressed miRNAs (DEMs) were predicted by TargetScan and miRanda. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of the predicted miRNA targets illustrated the putative roles for these DEMs in reproduction. The progress events were annotated, including oogenesis, lipid biosynthetic process, and related pathways such as apoptosis, ABC transporters, and amino acid metabolism. Four highly abundant DEMs (miR-9a-5p, miR-34-5p, miR-275-3p, and miR-317-3p) were further screened, and miR-34-5p was confirmed to be involved in the regulation of reproduction. Overexpression of miR-34-5p via injecting its mimics reduced fecundity and decreased Vg expression. Moreover, target genes prediction for miR-34-5p showed they might be involved in 20E signaling cascades, apoptosis, and gonadal development, including hormone receptor 4 (HR4), caspase-1 (Cp-1), and spermatogenesis-associated protein 20 (SPATA20). These findings provide a valuable resource for future studies on the role of miRNAs in BPH reproductive development.

The aims of this study were to explore the significant chemical changes in functional components induced by the traditional processing method and evaluate whether this method based on nine cycles of steaming and drying can effectively enhance the medicinal effects of Polygonatum cyrtonema rhizome. A global analysis on dynamic changes in secondary metabolites during nine processing cycles was performed, and the significantly differentially accumulated secondary metabolites were initially identified based on the secondary metabolome. Unsupervised principal component analysis (PCA), hierarchical clustering analysis (HCA), and orthogonal partial least squares discriminant analysis (OPLA-DA) on secondary metabolites clearly showed that processing significantly increased the global accumulation of secondary metabolites in processed P. cyrtonema rhizomes compared to unprocessed crude rhizomes. The first six processing cycles induced drastic changes in the accumulation of functional components, while the last three did not induce further changes. The accumulations of most functional components were significantly enhanced after the first three cycles and stabilized after six cycles; meanwhile, the first three cycles also led to numerous new components. However, the enhancing effects were unavoidably reversed or weakened under continued processing lasting 6–9 cycles. Furthermore, continued processing also reduced the contents of a small number of original components to undetectable levels. Processing induced some significantly enriched Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, among which the first three processing cycles enhanced the synthesis of various secondary metabolites and significantly affected the metabolisms of amino acids. In conclusion, this study not only reveals that processing can effectively enhance the medicinal effects, by two main mechanisms including enhancing chemical synthesis and inducing structural transformation of functional components, but also provides theoretical guidance for the optimization of the traditional processing method based on nine cycles of steaming and drying for achieving optimal effects on enhancing the medicinal effects of P. cyrtonema rhizome.

Purpose: In our previous study, exacerbation of albuminuria was observed in A1 adenosine receptor knockout (A1AR^−/-,) mice with diabetic nephropathy (DN), but the mechanism was unclear. Here, we investigated the relationship of megalin loss and albuminuria, to identify the protective effect of A1AR in megalin loss associated albuminuria by inhibiting pyroptosis-related caspase-1/IL-18 signaling of DN. Methods: We successfully collected DN patients’ samples and built diabetes mice models induced by streptozotocin. Megalin, cubilin, and A1AR expression were detected in kidney tissue samples from DN patients and mice through immunohistochemical and immunofluorescent staining. A1AR, caspase-1, interleukin-18 (IL-18) expression were analyzed using Western blotting in wild-type and A1AR^−/-, mice. Human renal proximal tubular epithelial cells (PTC) were cultured with high glucose to observe the effect of A1AR agonist and antagonist on caspase-1/IL-18 and megalin injury. Results: The loss of megalin, co-localized with A1AR at PTC, was associated with the level of albuminuria in diabetic patients and mice. The injury of megalin-cubilin was accompanied with the A1AR upregulation (1.30±0.1 vs 0.98±0.2, P=0.042), the caspase-1 (1.33±0.1 vs 1.0±0.2, P=0.036), and IL-18 (1.26±0.2 vs 0.96±0.2, P=0.026) signaling activation in mice with DN. More severe pathological injury, 24 hrs urine albumin excretion (170.8±4.1 μg/d vs 132.0±2.9 μg/d vs 17.9±2.8 μg/d, P<0.001) and megalin-cubilin loss were observed in A1AR^−/-, DN mice with more pronounced caspase-1 (1.52±0.03 vs 1.20±0.01, P=0.017) and IL-18 (1.42±0.02 vs 1.21±0.02, P=0.018) secretion. High glucose could stimulate the secretion of caspase-1 (1.72 times, P≤0.01) and IL-18 (1.64 times, P≤0.01), which was abolished by A1AR agonist and aggravated by A1AR antagonist. Conclusion: A1AR played a protective role in proximal tubular megalin loss associated albuminuria by inhibiting the pyroptosis-related caspase-1/IL-18 signaling in DN.

Entomopathogenic fungi (EPFs) can influence plant–insect interactions through complex molecular and chemical mechanisms. This study investigates how EPF treatment of tomato plants modulates volatile organic compound (VOC) emissions and subsequent trophic interactions between tomato plants, the herbivorous pest Phthorimaea absoluta, and the parasitic wasp, Trichogramma chilonis. Our results demonstrate that EPF-treated plants exhibited reduced attractiveness to adult P. absoluta moths, which were actively repelled by EPF-induced VOCs. Conversely, these same plants showed enhanced recruitment of the parasitoid T. chilonis, which demonstrated positive chemotaxis toward the modified VOC profile. Chemical analysis revealed significantly elevated emissions of key VOCs in EPF-treated plants, particularly (E)-β-Caryophyllene, β-phellandrene, and α-Phellandrene. This increase is correlated with enhanced production of defense-related phytohormones, including JA, SA, and JA-Ile, which may regulate VOC biosynthesis pathways. Behavioral response studies using synthetic VOCs and electroantennogram (EAG) measurements confirmed that these EPF-induced VOCs elicited strong olfactory responses in both insect species. To summarize, EPF treatment reshapes multitrophic interactions by strategically modulating plant VOC emissions and activating defense signaling pathways in tomato plants, providing new insights for potential applications in sustainable pest management strategies.