Chronic ultraviolet (UV) exposure drives skin degeneration, causing photoaging and increased carcinogenesis risk. To address complex pathogenesis and limited treatments, we developed GS5, a novel anti-photoaging sericin. GS5 fuses natural sericin with Seq10, a Kelch-like ECH-associated protein 1 (Keap1)-nuclear factor erythroid 2-related factor 2 (Nrf2) protein-protein interaction (PPI) inhibitor identified through molecular docking/dynamics. Seq10 binds Keap1, activates Nrf2 transcription, and alleviates UVA-induced photoaging Nrf2-dependently. Given the application bottlenecks of peptide molecules, we efficiently expressed the GS5 recombinant protein using the silk gland reactor of the silkworm and optimized the enzymatic extraction process to obtain high-activity GS5 sericin. In vitro, GS5 outperformed wild-type (WT) sericin and Seq10, enhancing viability/proliferation in irradiated keratinocytes and fibroblasts while reducing senescence markers (Senescence-associated β-galactosidase (SA-β-gal), P21), reactive oxygen species (ROS), DNA damage, and inflammation. GS5's photoprotection mechanistically requires Nrf2 activation. In vivo, GS5 reversed skin damage in UVA-irradiated mice, improving appearance and histology. RNA-seq implicated Janus kinase (JAK)-signal transducer and activator of transcription (STAT) pathway inhibition via immune/inflammation-related gene modulation. This study innovatively combines a targeted PPI inhibitor with sericin to create GS5, which mitigates photoaging through dual Nrf2 activation and JAK-STAT inhibition, offering a safe, effective, and sustainable therapeutic strategy.

Silkworm silk is a natural protein fiber composed mainly of fibroin and sericin, with protease inhibitors representing an additional abundant constituent. However, the impact of protease inhibitors on the structure and properties of silk fibers has not yet been studied. In this study, we focused on the Kunitz-type protease inhibitor SPI51, the most abundant protease inhibitors of cocoon. CRISPR/Cas9 editing was used to generate a homozygous mutant of SPI51 (SPI51KO), resulting in premature translation termination at the 33rd amino acid. After knocking out SPI51, the mechanical properties of silk were significantly reduced compared with those of the wild type. Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) results revealed that this deterioration was associated with significantly reduced β-sheet content and lower crystallinity. Morphological observations showed that the fibroin area of SPI51KO silk was significantly smaller than that in the wild type. Further Western blot analysis showed that fibroin heavy chain (Fib-H), fibroin light chain (Fib-L), and fibrohexamerin (P25) were decreased after knocking out SPI51, which resulted in a reduction of silk fibroin layer and affected structure and mechanical properties. Our results provide valuable insights into the balance between proteases and protease inhibitors in the silk gland and reveal for the first time the roles of the protease inhibitor in silk protein synthesis and the structural and mechanical properties of silk fibers.

Polarity serves as a vital parameter in assessing diagnosing diseases, reflecting quality in cellular metabolic status. In this work, two novel dicyanoisophorone-based fluorescence probes (DCIV-1 and DCIV-2) have designed and synthesized, capable of detecting pH levels and polarity fluctuation effectively. A detailed analysis of the probe's optical properties reveals that DCIV-1 and DCIV-2 exhibit distinct fluorescence and colorimetric responses under varying pH conditions. Meanwhile, DCIV-1 and DCIV-2 exhibit excellent responsiveness to polarity change respectively. Biological application reveals that both probes are successfully applied for distinguishing between cancer cells and normal cells. Furthermore, DCIV-1 and DCIV-2 enable rapid pH values in real water samples. As a convenient detection method, probe-based test strips have developed for detecting pH changes. This work highlights the potential of DCIV-1 and DCIV-2 for pH levels and polarity monitoring in biological and environmental fields.

Insects spin non-cocoon silk for protection, adhesion, and transfer with the environment. Sericin is a major component of non-cocoon silk fibers, yet our understanding of the sericin proteins is quite limited. In this study, we used CRISPR/Cas9-mediated gene editing to prepare a homozygous mutant strain that resulted in premature termination of the translation of non-cocoon sericin protein Ser5. We found that the silk glands of the Ser5 mutant (Ser5-/-) were smaller than those of the wild-type (WT), and both silk yield and major silk proteins significantly decreased in the larval stage. Seven kinds of non-cocoon silk were collected, and we found that the morphology of silk did not vary, but the diameter of the silk was significantly reduced in Ser5-/-. Further research revealed that the adhesive strength of native silk dope in Ser5-/- was significantly lower than that of the WT silkworm. Proteomic data indicated that the autophagy and apoptosis proteins increased significantly in Ser5-/-; differentially expressed proteins were enriched in pathways related to cellular stress responses and transcription and translation. Detection of the autophagy-related gene ATG8 also indicated that knockout of the Ser5 gene may lead to a level of cell stress, thus affecting the synthesis and secretion of silk proteins. Our study highlights the importance of the silk sericin gene in silk formation, silk protein adhesion, and the cellular developmental processes of the silkworm. These findings enhance our understanding of the functional roles of sericin genes in insects and provide a foundation for the development of sericin-based biomaterials.

Two structurally new benzene derivatives, designated as Guignarphenyl A (1) and Guignarphenyl B (2), and one structurally unique meroterpenoid named Guignarpenoid A (6) were successfully isolated and identified from the solid fermentation products of the endophytic fungus Guignardia mangiferae strain VDL166 (derived from Vaccinium dunalianum Wight, Ericaceae). Among these new compounds, compound 1 is reported for the first time in nature, while compounds 2 and 6 are newly identified analogs. Additionally, thirteen known secondary metabolites (3-5, 7-16) were obtained. The structures of all compounds were elucidated by comprehensive nuclear magnetic resonance (NMR) spectroscopic analysis combined with computed 13C NMR chemical shift analysis and DP4+ probability analysis for key relative configurations, and electronic circular dichroism (ECD) calculations. All isolates were evaluated for their in vitro antifungal activities against four phytopathogenic fungi (Alternaria brassicicola, Botrytis cinerea, Phytophthora infestans, and Valsa mali). Among them, compound 6 exhibited the most potent and broad-spectrum activity with MICs ranging from 12.5 to 25.0 µg/mL, comparable to the positive controls (Carbendazim and Thiabendazole). SUMMARY: Two new benzene derivatives, guignarphenyl A (1) and guignarphenyl B (2), and a novel meroterpenoid, guignarpenoid A (6), were isolated from Guignardia mangiferae VDL166. Structures were elucidated by NMR, ECD calculations, and DP4+ probability analysis. Compound 6 showed potent broad-spectrum antifungal activity (MICs 12.5-25.0 µg/mL). Bioactivity-guided study confirms the antifungal potential of G. mangiferae metabolites.

The utility of silk protein spans from East Asian traditions to promising applications in modern biomaterials science. While its bioactive effect is widely accepted, its molecular mechanisms and the supporting data have remained elusive. This study reveals the anti-photoaging mechanism of silk protein enzymatic hydrolysate (SEH), identifying key peptides SO1 from silk seroin and SC1 from silk sericin. Mechanistically, SO1 targets PDGFRβ to inhibit the NF-κB/ERK/SASP cellular senescence axis, while SC1 targets TRPV4 to suppress MITF-driven melanin synthesis. Additionally, the SO1 and SC1 combination suppresses key hallmarks of photoaging. It reduced senescence, confirmed by diminished β-galactosidase activity and SASP, while concurrently mitigating oxidative stress and hyperpigmentation, and promoting cell migration. Further network pharmacology analysis reveals a multi-component interaction network connecting bioactive silk peptides, target receptors, and downstream signaling pathways involved in mitigating photoaging. This integrative mechanism was defined as the “Silk Peptides Mesh”, with SO1 and SC1 identified as representative core peptides within this system. Finally, a 28-d skin clinical trial involving 30 participants confirms these findings, showing that a cream containing SO1 and SC1 improves photoaging-related skin indicators. Importantly, this work establishes a paradigm shift from conventional single-molecule approaches to a multi-component network understanding of bioactive peptide functionality.

The exceptional adaptability of insects to diverse food sources is central to their survival and evolutionary success. However, the molecular mechanisms underlying this rapid adaptation remain largely uncharacterized. In this study, adaptive phenotypic, transcriptomic, and metabolomic differences in silkworms fed mulberry leaves versus artificial diets were investigated. The results showed that dietary changes induced enrichment of multiple detoxification pathways in the fat body, midgut, and Malpighian tubules, accompanied by significant accumulation of secondary metabolites and xenobiotics such as flavonoids, terpenoids and saponins in these tissues. Stimulation experiments with nine upregulated metabolites in silkworm BmE cells revealed that most metabolites inhibited cell viability and induced detoxification genes such as GST, UGT and CYP upregulated, with flavonoids like genistein and daidzin exhibiting obvious inductive effects. Among the upregulated genes, GSTd2 frequently responded and was significantly upregulated in artificial diet-fed silkworms. Notably, overexpressing GSTd2 in BmE cells enhanced cell tolerance to genistein and daidzin. Furthermore, silkworms overexpressing GSTd2 showed higher flavonoid tolerance and better adaptability to artificial diets. In conclusion, this study provides valuable genetic targets for improving silkworm rearing efficiency on artificial diets, providing reference to optimize feed formulations and theoretical basis for understanding metabolic adaptation mechanisms to artificial diets in silkworms.

Cadmium ion (Cd²⁺) and phosphate hydrogen ion (HPO₄²⁻) are widely applied in our production and life, which are closely related to human health and environmental protection. Herein, leveraging the structural merits of isoborneol, a novel fluorescence probe IB-CdP has been designed and developed for sequential detection of Cd²⁺ and HPO₄²⁻. Upon interaction with Cd²⁺, IB-CdP exhibits a significant fluorescence enhancement at 530 nm, which is subsequently quenched by the addition of HPO₄²⁻. This probe achieves ultra-low detection limits (28.61 nM for Cd²⁺ and 30.89 nM for HPO₄²⁻) and rapid response times. In addition, practical applications results confirmed the reliability of probe IB-CdP in water samples analysis and cell imaging studies, demonstrating its extensive application in environmental monitoring and biomedical research.

DHX8 encodes a DEAH-box RNA helicase, an ATP-dependent enzyme that plays essential roles in RNA metabolism, including pre-mRNA splicing, transcription, and mRNA decay. Although DHX8 dysfunction has been linked with developmental abnormalities and disease pathogenesis in multiple model organisms, its biological functions in Lepidoptera, particularly in the silkworm Bombyx mori, remain unknown. To investigate the developmental role of B. mori DHX8 (BmDHX8), we generated knockout mutants using CRISPR-Cas9 genome editing. Genome sequencing confirmed frameshift mutations in the BmDHX8 locus. BmDHX8 mutants exhibited severe developmental defects such as dramatically reduced body size and premature lethality of silkworm larvae. Molecular characterization suggested systemic dysregulation, as evidenced by decreased triglyceride accumulation, impaired mTOR signaling activity, and increased aberrant splicing events. Therefore, these results indicate that loss of BmDHX8 is associated with aberrant splicing and alterations in lipid homeostasis and mTOR signaling pathways, potentially contributing to developmental defects. Taken together, our study offers an initial functional knockout analysis of BmDHX8 in regulating larval development in silkworms.

The study of volatile organic compounds (VOCs)-mediated plant growth promotion has long focused on various beneficial microbial species. As an important natural source of functional biomolecules, the biological function and potential value of VOCs released by plant pathogenic fungi in regulating plant growth still lack sufficient research, and further exploration is needed. In this study, a phytopathogenic fungus Alternaria alstroemeriae (strain Z84) was isolated from Vaccinium dunalianum for the first time, and the effects of its VOCs on the growth of Arabidopsis thaliana and Nicotiana benthamiana were systematically investigated. The results showed that after Z84 VOCs treatment, multiple phenotypic traits of the two plants were significantly improved, and the chlorophyll content was also markedly increased. Transcriptome analysis showed that a total of 1401 differentially expressed genes (DEGs) were identified in the treated A. thaliana, of which 629 were up-regulated and 772 were down-regulated. KEGG enrichment analysis showed that these DEGs were mainly enriched in photosynthesis-antenna proteins, plant-pathogen interaction, glutathione metabolism, plant hormone signal transduction, flavonoid biosynthesis and photosynthesis-related pathways. Metabolomics analysis revealed that Z84 VOCs treatment significantly changed the metabolic profile of A. thaliana, with the most significant changes in amino acid metabolism-related pathways. It is noteworthy that the plant hormone spectrum of A. thaliana was significantly changed after treatment, and the contents of salicylic acid (SA), abscisic acid (ABA) and gibberellins (GAs) were significantly up-regulated. These results not only demonstrate the potential of Z84-derived VOCs to facilitate plant growth but also provide an important basis for further dissecting the molecular mechanisms of plant-pathogenic fungi interactions.

BACKGROUND:Lepidopteran pests cause major agricultural losses, and overreliance on chemical pesticides raises environmental and health concerns. Genes controlling insect wing development offer promising targets for sustainable pest management. Here, we used the silkworm (Bombyx mori) as a model to investigate the transcription factor BmDll (BmDistal-less) in appendage development and to evaluate its potential as a target for RNA-based biopesticides. RESULTS:CRISPR/Cas9-mediated knockdown of BmDll caused severe defects in adult antennae, thoracic legs, and wings, including shortening, hardening, scale loss and complete wing malformation. Transcriptomic and quantitative real-time PCR (qPCR) analyses revealed that BmDll regulates cell differentiation and chitin assembly primarily through Hox and Notch signaling pathways. We developed chitosan (CS)/dsRNA (double-stranded RNA) nanoparticles targeting BmDll; oral delivery to silkworms and Spodoptera litura suppressed target gene expression and recapitulated the wing deformity phenotypes observed in the knockdown model. CONCLUSION:BmDll is a central regulator of lepidopteran appendage morphogenesis and a feasible target for dsRNA-based biopesticides. Nanoparticle-mediated delivery of BmDll-dsRNA provides a novel, environmentally friendly strategy to disrupt appendage development in pest insects. This work demonstrates the translation of fundamental developmental gene discovery into a potential agricultural application. © 2026 Society of Chemical Industry.

Nicotinamide adenine dinucleotide (NAD⁺) is a central metabolic coenzyme that regulates redox homeostasis, DNA repair, and cellular longevity. While the role of NAD⁺ metabolism in mammalian aging has been well studied, its significance in invertebrate systems remains underexplored. Here, we establish the silkworm (Bombyx mori) as a novel model for investigating NAD⁺-dependent lifespan regulation. Through phenotypic comparisons among silkworm strains, we found that longer-lived strains exhibit higher levels of NAD⁺ and elevated expression of BmNmnat1, a key enzyme in NAD⁺ biosynthesis. CRISPR/Cas9-mediated knockout and RNAi knockdown of BmNmnat1 led to embryonic lethality, increased DNA damage, disrupted cell cycle progression, and morphological aging phenotypes. Supplementation with nicotinic acid (NA) significantly reversed these aging-associated changes both in vitro and in vivo, including improved redox balance, reduced oxidative stress markers, and prolonged adult lifespan. Our results highlight the evolutionarily conserved role of BmNmnat1-mediated NAD⁺ metabolism in aging and establish the silkworm as a valuable invertebrate model for mechanistic studies and antiaging intervention screening.

As a key chemical raw material, hydrazine hydrate (N2H4) is widely used in chemical manufacturing, aerospace propulsion and pharmaceuticals synthesis. However, excessive N2H4 causes serious harms to the environment and organisms. Herein, a ratiometric fluorescence probe CDP has been designed and synthesized from p-hydroxycinnamic acid. Upon interaction with N2H4, the major fluorescence emission peak at 570 nm is blue-shifted to 455 nm, accompanied by a fluorescence color change from pale orange to blue. The detection limit of CDP for N2H4 is determined as 17.08 nM. Experimental data further indicate that probe CDP can effectively detect N2H4 across a broad pH range (5–12). More notably, probe CDP demonstrates significant potential for imaging N2H4 in living cells and mice models. Moreover, probe CDP can be employed for the detection of N2H4 in soil samples and water samples. This work describes a versatile fluorescence probe for detecting N2H4, underscoring its considerable potential in environmental analysis and public health monitoring.

Insect wing development involves tissue patterning, cell fate transitions, and hormone signaling, yet its spatiotemporal logic remains unclear. The silkworm, with large wing discs and defined stages, provides an ideal model for high resolution analysis. Here, we construct a spatiotemporal single-cell atlas of the silkworm wing disc across 10 timepoints, identifying 12 major cell types and their developmental transitions. Wing morphogenesis (Wm) cells act as central progenitors, differentiating into epithelial and cuticle lineages under lineage-specific transcription factors. Time‑resolved snRNA‑seq reveals hierarchical transcriptional reprogramming, with Wm cells functioning as early signaling hubs. Functional modules and signaling pathways were activated in spatiotemporal controlled manner. 20‑hydroxyecdysone treatment rapidly accelerates fate transitions and gene expression, recapitulating natural development within hours. Integration of morphology, hormone levels, and gene expression supports a five-stage Gene Transition Model describing progressive fate resolution. This work reveals wing development in silkworm and provides insights into hormone-driven organogenesis and potential manipulation of insect development in agriculture.

Two new hybrids of dehydrosterigmatocystin with a phenolic acid or a phenolic ether, designated aspergthones A (1) and B (2), together with four known compounds (3–6), were isolated from the EtOAc extract of Aspergillus versicolor VDL117, an endophytic fungus isolated from the leaf of Vaccinium dunalianum (Ericaceae). Specifically, aspergthone A (1) features dehydrosterigmatocystin linked to 2,4-dihydroxy-6-methylbenzoperoxoic acid, a compound also obtained from the same fungus, via an ester bond formed by dehydration condensation between the carboxyl group of the benzoperoxoic acid and a hydroxyl group of dehydrosterigmatocystin. In contrast, aspergthone B (2) is composed of dehydrosterigmatocystin conjugated to 3-(2,6-dihydroxy-4-methylphenoxy)-5-methylbenzene-1,2-diol through a similar condensation pathway. The planar structures and relative configurations were established by comprehensive spectroscopic analysis, and the absolute configurations of compounds 1 and 2 were determined by comparison of experimental and calculated electronic circular dichroism (ECD) spectra. In cytotoxicity assays, aspergthone B (2) showed moderate activity against paclitaxel-resistant human HepG-2 and SW480 cell lines, with IC₅₀ values of 26.99 ± 2.06 µM and 27.14 ± 1.63 µM, respectively, while the remaining compounds were inactive against both cell lines.

Intestinal microbiota are essential for insect nutrition and immunity, but how lepidopteran hosts sense and buffer microbial fluctuations remains poorly understood. Here we identify BmMBF2-11, a midgut-enriched multiprotein bridging factor 2 (MBF2) family protein in Bombyx mori, as a microbe-responsive factor that maintains gut homeostasis. RNA-seq of larval midguts from mulberry-fed (MF), artificial-diet-fed (AF), and Enterococcus mundtii-supplemented AF (AFE) groups, together with tissue expression analyses by qPCR, demonstrate that BmMBF2-11 is strongly induced by artificial-diet-associated dysbiosis and oral challenge with several opportunistic bacteria, and that its expression positively tracks intestinal bacterial burden. Using CRISPR/Cas9-mediated knockout and midgut-specific overexpression lines, together with 16S rRNA gene sequencing and infection assays, we demonstrate that BmMBF2-11 limits total bacterial load, preserves α-diversity and community structure under both mulberry and artificial-diet conditions, and promotes the inducible expression of multiple antimicrobial peptide genes in response to E. mundtii and Staphylococcus sciuri. Loss of BmMBF2-11 accelerates the expansion of E. mundtii, impairs bacterial clearance, and reduces host survival after oral infection. Our findings identify BmMBF2-11 as an MBF2-domain protein that converts microbial perturbations into appropriately scaled antibacterial responses, thereby contributing to the maintenance of gut microbiota homeostasis in Bombyx mori.

Four lignans (1–4), including a new one, namely (7R,8S)-9-hydroxy-3,4-methylenedioxy-3′,9′-dimethoxy[7-O-4′,8-5′]neolignan (1), were isolated from the ethanolic extract of the dried leaves of Phyllanthus emblica L. Their structures were determined on the basis of extensive MS, 1D and 2D NMR spectroscopic techniques, and comparison with literature data. Compounds 1–4 exhibited low sensitivity towards five specific human cancer cell lines and no obvious antioxidant activity.

Centromere protein J (CENPJ) is a core centrosomal protein essential for centriole duplication and microtubule organization, and mutations in the gene cause aberrant neuronal morphology and primary microcephaly in mammals. However, its functions in insect development remain poorly understood. In this study, we investigated the role of CENPJ in the silkworm Bombyx mori, a lepidopteran model featuring unique holocentric chromosomes. We found that CENPJ is evolutionarily conserved and predominantly expressed in silkworm gonads. Cellular assays demonstrated that CENPJ exhibits cytoplasmic localization and regulates cell proliferation and DNA replication. CRISPR-Cas9-mediated knockout of CENPJ caused almost sterility in both sexes, leading to severe defects in ovarian and testicular development, impaired formation of both eupyrene and apyrene sperm, and abnormal structures of the spermatheca and seminal vesicle. Transcriptional analysis revealed downregulation of genes involved in cell cycle and gonad development in mutant individuates. These findings establish CENPJ as a master regulator of insect reproductive development and highlight its potential as a molecular target for reproductive disruption in lepidopteran pest management.

Cadmium ion (Cd2+) and phosphate hydrogen ion (HPO42-) are widely applied in our production and life, which are closely related to human health and environmental protection. Herein, leveraging the structural merits of isoborneol, a novel fluorescence probe IB-CdP has been designed and developed for sequential detection of Cd2+ and HPO42-. Upon interaction with Cd2+, IB-CdP exhibits a significant fluorescence enhancement at 530 nm, which is subsequently quenched by the addition of HPO42-. This probe achieves ultra-low detection limits (28.61 nM for Cd2+ and 30.89 nM for HPO42-) and rapid response times. In addition, practical applications results confirmed the reliability of probe IB-CdP in water samples analysis and cell imaging studies, demonstrating its extensive application in environmental monitoring and biomedical research.