BACKGROUND:Pseudomonas aeruginosa (P. aeruginosa) is a common opportunistic pathogen responsible for skin infections. Tannic acid, a representative hydrolysable tannin, has broad-spectrum antibacterial properties and potential applications in wound healing. OBJECTIVE:To characterize the concentration-dependent antibacterial and anti-virulence effects of TA against P. aeruginosa, identify associated transcriptional responses, and evaluate its therapeutic potential in an infected wound model. RESULTS:High concentrations of tannic acid disrupted the integrity of the P. aeruginosa cell membrane, leading to leakage of intracellular components, and were associated with marked alterations in bacterial protein profiles. At sub-inhibitory concentrations (sub-MICs), tannic acid significantly attenuated bacterial virulence by suppressing pyocyanin and rhamnolipid biosynthesis, biofilm formation, proteinase activity, and bacterial motility. TA treatment at 64 μg/mL was associated with transcriptional changes in genes involved in biofilm formation and efflux functions. In a rat wound model infected with P. aeruginosa, tannic acid treatment significantly decreased bacterial burden, attenuated histopathological injury, contained infection progression, and expedited wound closure. CONCLUSION:This study evaluated the antibacterial and anti-virulence effects of tannic acid against P. aeruginosa, characterized associated transcriptional responses using RNA sequencing, and assessed its therapeutic potential in an infected wound model.

To elucidate the anti-rheumatoid arthritis (RA) mechanisms of Notopterygium incisum essential oil (NIEO) using an integrated strategy combining chemical profiling, network pharmacology, multi-omics data mining, and advanced machine learning, while developing an optimized nanoemulsion (NE) delivery system to enhance its therapeutic applicability. Gas chromatography-mass spectrometry was employed for chemical profiling of NIEO. Multi-omics integration and machine learning models were used to identify core therapeutic targets, with diagnostic potential assessed via receiver operating characteristic analysis. Computational modeling, including molecular docking and dynamics simulations, provided structural insights into NIEO components interacting with targets. An optimized NIEO-NE was developed and characterized for physicochemical properties [mean diameter, polydispersity index (PDI), encapsulation efficiency]. In vitro assays evaluated NIEO-NE effects on lipopolysaccharide-induced Caspase-1 (CASP1) expression, reactive oxygen species (ROS) production, and pro-inflammatory mediators including interleukin (IL)-6, tumor necrosis factor (TNF)-α, IL-1β, nitric oxide (NO) in RAW264.7 macrophages. GC-MS identified a-terpineol as the predominant constituent among 17 primary volatile compounds in NIEO. CASP1 was identified as the core target, with an area under curve of 0.985 for RA diagnosis. Molecular docking and dynamics revealed strong interactions between key NIEO components and CASP1’s active site. The optimized NIEO-NE exhibited a mean diameter of 82.3 nm, PDI of 0.12, and encapsulation efficiency of 94.8

Acute kidney injury (AKI) lacks targeted pharmacological interventions. While the DanShen-DaHuang (DS-DH) herb pair shows clinical potential for AKI treatment, and our prior study has validated its nephroprotective efficacy in a cisplatin-induced murine model, its specific molecular targets within the renal microenvironment remain undefined. In this study, we integrated network pharmacology and weighted gene co-expression network analysis (WGCNA) to screen AKI-related targets of the DS-DH pair. A multi-algorithmic machine learning pipeline (including LASSO, Boruta, Random Forest, GBM, XGBoost, and Decision Trees) was utilized to calculate feature importance scores and rank core genes. Subsequently, single-cell RNA sequencing (scRNA-seq) data (GSE197266) were analyzed for transcriptomic mapping, pseudotime trajectory, and cell-cell communication. Finally, molecular docking evaluated theoretical binding affinities. After database screening, a total of 603 drug-disease intersecting targets were obtained. Subsequently, 917 module genes significantly associated with AKI were identified by WGCNA, and 62 core candidate genes were determined after intersecting with the above targets. Multi-algorithm machine learning ranked the importance of the 62 targets, with transketolase (TKT) ranking the highest. To elucidate the mechanism of TKT in AKI, scRNA-seq analysis was performed on 77,593 high-quality cells. The results showed that Tkt was specifically enriched in renal macrophages, with the highest expression in the M2-polarized subset. Pseudotime analysis further revealed that Tkt expression dynamics were highly synchronized with the differentiation trajectory of M2 macrophages and positively correlated with the repair markers Arg1 and Mrc1. Cell-cell communication analysis predicted that Tkt+ M2 macrophages act as active communication hubs via the Spp1 and Mif signaling axes. Molecular docking validated the favorable binding affinity between core DS-DH compounds and the TKT active pocket. This computational framework predicts that the DS-DH herb pair might mitigate AKI by potentially targeting TKT, a metabolic enzyme closely associated with macrophage M2 polarization. By prioritizing targets via multi-algorithmic scoring, we provide a data-driven rationale and candidate targets for future experimental validation.

The main objective of this study was to preliminarily analyze the major flavonoid and phenolic acid components of the ethanolic extract of Gerbera delavayi Franch (E-GDF), and to evaluate its anti-inflammatory and antioxidant properties in lipopolysaccharide (LPS)-stimulated murine macrophage RAW264.7 cells and systemic inflammation mouse models. Results indicated that E-GDF was rich in flavonoids (16.35 ± 0.19 mg RT/g d.w. Plant Material) and polyphenolic compounds (36.15 ± 0.20 mg GAE/g d.w. Plant Material). LC-MS analysis of E-GDF revealed that its major flavonoid components included kaempferol glycosides, luteolin, and their glycosylated derivatives, while its phenolic acids were predominantly chlorogenic acid, caffeic acid, ferulic acid, and their corresponding glycosides. E-GDF exhibited good antioxidant activities, including the scavenging of DPPH, ABTS, •OH, and O2•- radicals. E-GDF treatment significantly inhibited the production of ROS and inflammatory mediators (NO, IL-6, TNF-α) in LPS-stimulated macrophages (RAW 264.7), while concurrently down-regulating the mRNA expression of COX-2, IL-1β, Casp1, and GSDMD-1. In addition, in vivo experiments revealed that E-GDF treatment effectively reduced the serum LPS, AST levels, as well as hepatic TNF-α, IL-6 levels in mice with LPS-induced acute liver injury. Furthermore, E-GDF significantly ameliorated LPS-induced liver pathological damage. These results provide a basis for G. delavayi as a potential antioxidant, anti-inflammatory, and hepatoprotective herbal medicine.

Acute kidney injury (AKI) presents a critical clinical challenge due to its rapid progression and lack of effective targeted therapies. The herbal combination of rhubarb and Salvia miltiorrhiza, a cornerstone of Traditional Chinese Medicine (TCM) for renal protection, shows promise, yet its bioactive components and mode of action remain incompletely understood. This study identifies and characterizes inherent nanoscale entities from this herbal pair as a novel nanotherapeutic platform. Self-assembled nanoparticles (designated RSNPs) were isolated from the ethanol extract via differential centrifugation. Comprehensive characterization revealed that RSNPs form stable nanostructures through spontaneous self-assembly, primarily driven by supramolecular interactions (e.g., π-π stacking and hydrogen bonding). UPLC-MS/MS quantification confirmed the co-assembly of multiple bioactive constituents within RSNPs. Network pharmacology and molecular docking initially predicted their synergistic action on AKI-related pathways. In a cisplatin-induced murine AKI model, RSNP administration markedly attenuated renal dysfunction and histopathological damage, mechanistically linked to the mitigation of oxidative stress (e.g., decreased MDA and increased SOD) and inflammation (e.g., downregulated TNF-α and IL-6). In vitro, RSNPs demonstrated enhanced cellular internalization and superior cytoprotection against cisplatin toxicity in renal tubular epithelial cells, significantly reducing apoptosis. These findings unveil that the therapeutic efficacy of the Rheum palmatum L.-Salvia miltiorrhiza Bunge pair is intrinsically embedded within its nanoscale architecture. RSNPs represent a new class of TCM-derived nanotherapeutics with a well-defined material basis and multimodal mechanisms, offering a promising strategy for AKI treatment.

Trichophyton verrucosum is a zoophilic dermatophyte and the primary cause of bovine dermatophytosis, a contagious and economically significant skin disease with zoonotic potential. Despite its importance, high-quality genomic and transcriptomic resources for this species remain scarce, limiting insights into its pathogenic mechanisms and antifungal responses. Here, we present a chromosome-level genome assembly of T. verrucosum OR056436.1, generated using single-molecule real-time (SMRT), Illumina paired-end and high-throughput chromosomal conformation capture (Hi-C) technologies, followed by comprehensive annotation and comparative analyses with six other fungal pathogens. Transcriptome profiling under itraconazole (ITR) and Xuanjing - Compound Chinese Herbal Spray (CCM) treatments revealed antifungal-responsive genes. The final 23.44 Mb genome comprises 7,936 protein-coding genes, with 4.98% repetitive sequences, and shows 98.8% completeness based on BUSCO analysis. Functional annotation identified 262 Kyoto Encyclopedia of Genes and Genomes pathways, 4,861 Eukaryotic Orthologous Groups proteins, 34 secondary metabolite biosynthetic gene clusters, 1,157 carbohydrate-active enzymes, 2,920 pathogen-host interaction genes, and 59 virulence factor genes. Comparative genomics identified 45,425 orthogroups including 393 core and 184 single-copy orthogroups, placing T. verrucosum closest to Trichophyton rubrum. Transcriptomic analysis highlighted differentially expressed genes enriched in amino acid metabolism, ABC transporters, drug metabolism, and stress response pathways. Notably, subtilisin-like proteases and LysM-domain proteins were significantly regulated under antifungal treatments. This integrative genomic-transcriptomic study provides the most complete T. verrucosum genome to date, identifies key genes involved in pathogenicity and antifungal adaptation, and offers valuable resources for diagnostics, therapeutic development, and improved strategies for bovine dermatophytosis.

Mycoplasma synoviae (MS) is a prevalent pathogen in poultry farming, often causing infectious synovitis and arthritis in poultry, resulting in joint swelling, lameness, and growth retardation, thereby significantly impacting production efficiency. Tengchuan compound mixture (TCM), a traditional Chinese medicine formulation designed to against MS infection effectively, but its active components and mechanisms isn't elucidated. This study utilized liquid chromatography-mass spectrometry (LC-MS) to identify the active components in TCM and employed network pharmacology to predict key targets and pathways in against MS infection. A model of MS infection was established in specific-pathogen-free (SPF) chickens to assess the pharmacodynamic effects of TCM on parameters such as body weight, clinical manifestations, histopathology of tarsal joints, and serum inflammatory markers. Metabolomics analysis and 16S rDNA sequencing were conducted to elucidate the potential mechanism of TCM in preventing and treating MS infection. Western blot analysis confirmed the core targets identified by network pharmacology. TCM treatment effectively mitigated MS-induced joint lesions by suppressing the phosphorylation of proteins associated with the JAK2-STAT3 and PI3K-AKT signaling pathways. Pharmacokinetic analysis identified 18 major potential active ingredients. Network pharmacology identified 147 common targets of TCM against MS infection, including 13 core targets such as SRC, JAK2, PIK3CA, and STAT3. Metabolomics analysis revealed that MS infection disrupted amino acid and energy metabolism in chickens, which was partially restored by TCM treatment. Additionally, 16S rDNA sequencing indicated that TCM could recovery the gut microbiota disrupted by MS infection. In conclusion, this integrated multi-omics study provides mechanistic insights into TCM's effects during MS infection, highlighting its role in ameliorating infection-induced inflammation and joint pathology while modulating host metabolic pathways and gut microbiota. These findings offer a comprehensive understanding of TCM's therapeutic potential for managing Mycoplasma synoviae infection.

Background/Objectives: Blood is an essential component of the immune system. As post-transcriptional regulators, miRNAs, abundant in blood, are necessary aspects in blood’s immune and physiological functions. However, there is limited knowledge about the expression and function of miRNAs in the blood of giant pandas. Methods: We comparatively analyzed miRNA expression profiles in the blood of giant pandas of different ages using small-RNA sequencing technology. Results: We identified 393 known miRNAs, 219 conserved miRNAs, and 71 novel miRNAs in the blood of giant pandas, and functional enrichment analysis showed that the genes regulated by DE (differentially expressed) miRNAs were mainly enriched in the regulation of enzyme-linked receptor protein signaling pathways and the signaling pathways of MAPK, Hippo, and FoXO. Conclusions: Our study clarified giant pandas’ blood miRNA expression profiles at different developmental stages, which will help elucidate the blood immunity and regulation of blood cell physiological functions in giant pandas.

Pseudorabies virus (PRV), a swine alphaherpesvirus, is a double-stranded DNA virus. It may infect various animals, especially pigs. PRV infection in pigs leads to high mortality rates, and causes huge economic lose for swine industry. Currently, there are few effective antiviral treatments available. Rosmarinic acid (RA), a hydrophilic phenolic compound, shows potential for inhibiting herpes simplex virus. Given that PRV is a member of the Herpesviridae family, this study investigated the antiviral effects of RA against PRV infection through both in vitro and in vivo, as well as the underlying molecular mechanisms. PK-15 cells were used to assess the cytotoxicity of RA in vitro, followed by an investigation of its anti-PRV activity. The study then explored how RA regulates the cGAS-STING signaling pathway, along with inflammatory and apoptotic factors in PRV-infected cells. Molecular docking and dynamics simulations further elucidated the binding interactions between RA and cGAS-STING, providing insight into how RA activates the cGAS-STING pathway against PRV infection. In vivo, the antiviral efficacy of RA was evaluated in a PRV-infected mouse model by assessing tissue viral genome copies, the innate immune cGAS-STING signaling pathway activation, and inflammatory and apoptotic responses. The results showed that RA exhibited a half-maximal cytotoxic concentration (CC50) of 26.23 µg/mL on PK-15 cells and a half-maximal inhibitory concentration (IC50) of 0.84 µg/mL against PRV, resulting in a selectivity index (SI) of 31.22. These findings suggest that RA is a highly effective and low-toxicity compound. RA significantly inhibited PRV adsorption, penetration, and replication within cells. Additionally, while PRV infection suppresses the cGAS-STING signaling pathway, RA treatment activates the innate immune response, enhances downstream antiviral effector IFN-β expression, and reduces inflammation and apoptosis in PRV-infected cells. Molecular docking results showed that the docking scores of cGAS_RA and STING_RA complexes were both less than − 5 kcal/mol, suggesting that RA binds well to cGAS and STING proteins. Molecular dynamics simulations, including RMSD, RMSF, and MM-GBSA analyses, confirmed the high binding stability of cGAS with RA, further validating the potential activity of RA as a cGAS agonist. In vivo studies revealed that RA dramatically lowered viral genome copies in various organs, activated the cGAS-STING signaling pathway, inhibited PRV-induced inflammation and apoptosis, alleviated clinical symptoms, and decreased mortality rate in PRV-infected mice. Overall, RA significantly inhibited PRV proliferation in vitro and in vivo, effectively reduced inflammation and apoptosis, and decreased the mortality rate in infected mice. The study supports the development of RA as an antiviral drug and emphasizes its potential as a candidate for PRV therapy.

Background: This study investigates the neuroprotective effects of 1,8-cineole (1,8-CH), against hippocampal oxidative stress in a chronic unpredictable mild stress (CUMS) mice model of depression, focusing on the underlying molecular mechanisms. Methods: The effects of CUMS exposure were assessed by measuring oxidative stress markers, antioxidant activity, and neuronal damage in the hippocampus using histopathology, network pharmacology, Western blot analysis, and small interfering RNA (siRNA) knockdown experiments. Results: 1,8-CH significantly alleviated depression-like behaviors in CUMS mice. CUMS exposure induced oxidative stress in the hippocampus, evidenced by elevated MDA levels, decreased antioxidant activity, and neuronal damage. DHE staining revealed ROS accumulation. Treatment with 1,8-CH alleviated oxidative stress by reducing MDA, restoring antioxidant activity, and lowering ROS levels, while improving neuronal structure. Network pharmacology identified the PI3K/Akt/Nrf2 pathway as a key mediator of 1,8-CH’s neuroprotection, which was supported by Western blot results, demonstrating PI3K/Akt activation and a potential enhancement of Nrf2 nuclear translocation. Furthermore, in corticosterone-induced PC12 cells, the antioxidant effects of 1,8-CH were abolished by Nrf2 inhibition and siRNA knockdown, confirming Nrf2’s role. Conclusions: These findings suggest that 1,8-CH alleviates hippocampal oxidative stress in CUMS-induced depression via the PI3K/Akt/Nrf2 pathway, highlighting its potential as a health supplement for managing depression.

Pseudorabies virus (PRV) can infect most mammals and has caused significant economic losses in global pig production. The emergence of new mutants significantly reduces the protective effect of vaccination, indicating an urgent need for the development of specific therapeutic agents against PRV infection. In this study, we analyzed the changes in the cellular proteome after PRV infection in resveratrol-treated PK-15 cells using TMT quantitative proteomics combined with LC-MS/MS. The results identified the differential proteins osteopontin (iOPN) and interleukin-1 receptor accessory protein (IL-1RAP), which have significant biological implications. The regulation of OPN-IL-1β signaling by PRV infection was further studied through the OPN-ERK/JNK-IL-1β signaling axis. The transcriptional levels of OPN, C-JUN, IL-1RAP, and IL-1β, along with the protein levels of ERK, JNK, C-Jun, and their phosphorylated forms at 8, 12, and 16 h post-infection, were determined. The results showed that PRV infection inhibited the activation of this signaling axis, which was upregulated by resveratrol treatment. Down-regulation of OPN by siRNA increased PRV proliferation and inhibited the activation of the signaling axis, which was antagonized by resveratrol treatment. In PRV-infected mice, resveratrol treatment produced the same changes observed in vitro. The present study demonstrated that resveratrol can promote innate immune responses by regulating the OPN-ERK/JNK-IL-1β signaling axis, thereby activating host antiviral defenses against PRV infection. SIGNIFICANCE: Resveratrol targets the OPN-ERK/JNK-IL-1β axis to enhance innate immunity, offering a novel antiviral strategy against PRV infection. This study identifies OPN as a key regulator of host defense, linking ERK/JNK signaling to IL-1β-mediated antiviral responses. In vivo validation demonstrates resveratrol's therapeutic potential, reducing PRV replication and mortality in mice via immune pathway activation.

Parishin C (PaC) is an active ingredient in Gastrodia elata Bl. that has neuroprotective effects. However, research on its role in oxidative stress and neuroinflammation is still limited. This study used LPS–stimulated HT22 cells to investigate the antioxidant properties of PaC. Through the co–culture system of HT22 and BV2 cells, the effect of PaC on neuroinflammation was explored. The current results indicated that PaC can inhibit the levels of reactive oxygen species and peroxides in LPS–stimulated HT22 cells and increase the levels of antioxidant factors. Meanwhile, PaC can also inhibit neuronal ferroptosis and the levels of pro–inflammatory cytokines in BV2 cells. Importantly, the antioxidant and anti–inflammatory effects of PaC are achieved by activating the Nrf2 signaling pathway. The WB and IF results indicated that PaC can promote nuclear translocation of Nrf2, activate downstream antioxidant factors, and thereby regulate inflammatory responses. Inhibition of Nrf2 can significantly inhibit the regulation of PaC on the Nrf2 signaling pathway. These results indicated that PaC can activate the Nrf2 signaling pathway to inhibit oxidative stress and inflammation.

The P. aeruginosa, widely spread and highly detrimental nosocomial pathogen, exhibits extensive intrinsic and acquired antibiotics resistance. Naringenin, a flavonoid compound, exhibits diverse pharmacological activities, especially in inhibiting the growth of many pathogenic bacteria. Our study revealed that naringenin showed significant inhibition on the virulence factors of P. aeruginosa at non bactericidal concentrations. At the concentration of 200 μg·mL−1, naringenin inhibited the swarming and swimming motility of P. aeruginosa, reduced pyocyanin secretion, and flagella assembly. Quantitative real-time polymerase chain reaction experiment was conducted, and the results showed that naringenin could reduce the expression of bacterial virulence factors related to P. aeruginosa. Naringenin inhibited bacterial adherence capability to cells in vitro. The larvae of Galleria mellonella showed a higher survival rate in treatment with naringenin in vivo. These findings suggested a promising role of naringenin in suppressing the virulence of P. aeruginosa, which encompasses the growth inhibition of various pathogenic bacteria. This indicates substantial potential for future therapeutic interventions targeting bacterial virulence.

To reveal the molecular regulatory differences in physiological adaptations of pigeons with different breeding objectives, this study compared the transcriptomic differences in the hearts and cerebellar vermis of homing pigeons (HP) and meat pigeons (MP). Phenotypic analysis showed that the heart index of HP was significantly higher than that of MP, which helps enhance functions such as blood circulation, supporting the metabolic demands for long-duration activity. However, no significant differences were observed in the cerebellar vermis indices. Transcriptomic analysis identified 162 differentially expressed genes (DEGs) in the heart and 192 in the cerebellar vermis. Differential genes in the heart were primarily enriched in pathways related to circulatory system processes, ventricle development, fatty acid metabolism, and hypoxia response, including key genes related to angiogenesis and metabolism, such as HSP90AA1, ACACB, and LPIN1, suggesting that HP have stronger cardiovascular function and energy metabolism capacity. Differential genes in the cerebellar vermis were enriched in extracellular matrix organization, synaptic organization, axon guidance, and NRF2 pathway, indicating enhanced cerebellar neural adaptability and oxidative stress resistance in HP. Notably, circadian rhythm-related genes (PER2, PER3) were downregulated in both the hearts and cerebellar vermis of HP, which may affect the adaptive growth and remodeling of the heart. Alternative splicing analysis revealed 149 and 151 significant splicing events in the heart and cerebellar vermis, respectively, with exon skipping being the predominant splicing type. These results systematically reveal the tissue-specific molecular characteristics of the heart and cerebellar vermis in MP and HP, providing new insights into the molecular basis of their cardiac and metabolic adaptation.

Background and aimsPolygonum cuspidatum Sieb.et Zucc. (P. cuspidatum) and its active components have been clinically proven to have anti-hepatocellular carcinoma effects. However, the potential targets of P. cuspidatum for these effects have not yet been revealed.MethodsWe used network pharmacology and single-cell transcriptomic analysis with molecular docking to elucidate the active components and targets of P. cuspidatum for hepatocellular carcinoma.ResultsCDK1, ESR1, HSP90A11, and MAPK1 were shown to be the key targets of P. cuspidatum for hepatocellular carcinoma. P. cuspidatum was found to be likely correlated with the improved abnormal expression of CDK1 and ESR1 and the poor prognosis of HSP90AA1 and MAPK1. CDK1 was identified as the most potential anti-hepatocellular carcinoma target of P. cuspidatum. Among the active components of P. cuspidatum, physcion diglucoside was found to have the most potential to treat hepatocellular carcinoma by targeting CDK1.ConclusionOur study provides novel insights into the anti-hepatocellular carcinoma pharmacological effects of P. cuspidatum, which could serve as a scientific basis for its development as a medicinal resource and the targeting of CDK1 for hepatocellular carcinoma treatment.

Background and aims:To identify biomarkers to predict acute liver failure and investigate the mechanisms and immune-related pathways linked to its onset and progression. Methods:We analyzed gene expression differences between patients with acute liver failure(ALF)and controls in the GSE14668 dataset.Clinically relevant modules and key ALF-associated genes were identified using weighted gene co-expression network analysis(WGCNA)in conjunction with differential gene expression(DEG)analysis.Enrichment analysis was carried out and protein-protein interaction networks were constructed to understand the functions and pathways.Six potential diagnostic biomarkers were identified using machine learning algorithms.Diagnostic performance was assessed via column charts and area under the curve calculations.Single-sample gene set enrichment analysis evaluated the relationship between known marker gene sets and potential biomarker expression.We also examined diagnostic biomarker mRNA levels in ALF models in vivo and in vitro.We estimated the relative infiltration levels of 22 immune cell subpopulations in ALF samples,and explored the link between diagnostic biomarkers and infiltrating immune cells. Result:We found 352 DEGs associated with ALF.WGCNA analysis and intersecting DEGs identified 191 significant ALF-related genes.Machine learning identified HORMAD2,WNT10A,ATP6V1E2,CMBL,ARRDC4,and LPIN2 as potential diagnostic biomarkers.Cell experiments and quantitative real-time polymerase chain reaction supported the therapeutic potential of eriodictyol for ALF.Immune infiltration analysis suggested that plasma cells,CD4 memory resting and activated T cells,macrophages,and neutrophils might play roles in the progression of ALF. Conclusion:We identified HORMAD2,WNT10A,ATP6V1E2,CMBL,ARRDC4,and LPIN2,as diagnostic biomarkers for ALF and demonstrated the effectiveness of eriodictyol for treating ALF.Immune cell infiltration may play a significant role in the pathogenesis and progression of ALF.

Naringenin is a natural dihydro-flavonoid compound with various beneficial pharmacological activities. However, the extremely poor water solubility and bioavailability pose significant challenges for its application. In this study, chitosan-casein-naringenin nanoparticles (Cs-cas-Nar) were prepared to enhance naringenin's water solubility and bioavailability. Cs-cas-Nar had a particle size of 225.8 nm, a PDI of 0.155, a zeta potential of 27.2 mV, and an encapsulation efficiency of 87.02 %. FT-IR analysis indicated that casein encapsulated naringenin through hydrogen bonds and non-covalent interactions. Simulated gastrointestinal digestion and in vivo release results demonstrated that Cs-cas-Nar significantly improved naringenin's water solubility and bioavailability. The sitecontrolled and rapid release of Cs-cas-Nar provided higher drug concentration in the intestine and lung tissue. In in vivo experiments, Cs-cas-Nar provided a protective effect against Pseudomonas aeruginosa-induced pneumonia in mice. Cs-cas-Nar not only reduced weight loss and lung bacterial load of pneumonia mice, but also reduced the production of pro-inflammatory factors IL-1 beta and IL-6 by >50 % and restored the antioxidant capacity of the lungs. Through computer simulation technology and qPCR detection, we confirmed that Cs-cas-Nar can inhibit the assembly of P. aeruginosa flagella. These findings suggest that Cs-cas-Nar is a simple and easily manufactured formulation with potential application value in treating P. aeruginosa infection.

The rapid emergence of antibiotic-resistant strains of Staphylococcus aureus presents a substantial challenge to global public health, underscoring the urgent need for novel antibiotics with diverse mechanisms of action. In this study, we conducted mutagenesis on the C-terminal region of the lantibiotic ripcin C to enhance its antimicrobial efficacy against S. aureus. The resulting optimized variant, ripcin CP23A, demonstrated potent and selective antimicrobial activity, with a minimal inhibitory concentration of 2-4 mg/L against S. aureus. Beyond its strong antimicrobial properties, ripcin CP23A exhibited significant antibiofilm activity against methicillin-resistant S. aureus (MRSA). Mechanistic studies revealed that, in addition to targeting lipid II, ripcin CP23A disrupts bacterial membranes, a capability absent in ripcin C, which may contribute to its superior antimicrobial and antibiofilm effects. Moreover, ripcin CP23A displayed favorable biosafety and plasma stability profiles. Notably, in a mouse model of MRSA-induced mastitis, ripcin CP23A effectively reduced bacterial load, alleviated inflammation, and preserved the normal histomorphology of mammary glands. This study introduces ripcin CP23A as a promising antibiotic candidate for the treatment of MRSA-related infections.