Breast cancer (BRCA) is the most common cancer worldwide, with an incidence exceeding that of lung cancer. Protein lactylation, a newly identified post-translational modification involving the binding of lactic acid to lysine residues, plays an important role in BRCA. However, its role in BRCA progression remains largely unexplored. This study aims to identify and characterize the lactylation-related genes involved in BRCA biology. Transcriptomic and clinical data of BRCA and normal breast tissues were obtained from TCGA and GEO. Lactylation-related genes were curated from literature and intersected with BRCA datasets to identify candidates. A prognostic risk model was constructed using LASSO and Cox regression. Functional enrichment was performed using KEGG, GSVA, and GSEA. Immune correlations were evaluated by ESTIMATE, CIBERSORT. Single-cell RNA-seq data were integrated to assess gene expression heterogeneity across tumor and immune compartments. In vitro, MDA-MB-231 cells were treated with sodium L-lactate and lactylation-inducing agents, and gene expression was validated by Western blot and RT-qPCR, while EdU and wound healing assays evaluated proliferation and migration. We identified six hub genes associated with the immune microenvironment. Notably, S100A4 is significantly underexpressed, suggesting their potential regulatory roles in BRCA. Further analysis demonstrated that lactylation-related genes are closely linked to immune regulation in BRCA, indicating a possible crosstalk between metabolic modification and tumor immunity. Additionally, we found that lactylation significantly influences gene expression patterns and immune infiltration in BRCA. Importantly, lactic acid ions were shown to upregulate lactylation levels in BRCA cells, underscoring the functional impact of metabolic signals on post-translational modifications in tumorigenesis. Our findings indicate a potential mechanism wherein lactylation affects BRCA progression via lactic acid-driven regulation of the immune microenvironment; they also highlight the possible involvement of S100A4 in this process and offer new insights that could contribute to the diagnosis and treatment of BRCA.

Sweet-waxy maize is a highly valuable specialty maize type with an increasing market demand, but conventional breeding methods for producing sweet-waxy maize are restricted by severe bottlenecks, such as long breeding cycles and linkage drag. This study was conducted to rapidly create sweet-waxy maize germplasm using CRISPR/Cas9 genome-editing technology. We used a CRISPR/Cas9 system to target maize Sh2 (regulating the super-sweet kernel trait) and Wx (controlling the waxy kernel trait), which are two key genes in the starch biosynthesis pathway. Two small-guide RNAs (sgRNAs) designed for each gene were incorporated into CRISPR/Cas9 vectors, which were then introduced into maize via Agrobacterium-mediated transformation. We obtained Cas9-free T3 homozygous sh2 and wx mutant lines with significant increases in kernel soluble sugar and amylopectin contents, respectively, but no adverse changes to major agronomic traits. Using these Cas9-free lines, we developed a new type of sweet-waxy maize germplasm, in which waxy and sweet kernels on the same ear segregated at a 3:1 ratio. Our results indicate that CRISPR/Cas9-mediated editing of Sh2 and Wx can efficiently generate sweet-waxy maize germplasm with no detectable linkage drag. The study methods would be useful for optimizing the molecular breeding of novel and innovative maize germplasm.

Excessive osteoclast activity drives inflammatory bone loss in osteoporosis, rheumatoid arthritis, and periodontitis. Natural compounds represent promising therapeutic candidates with favorable safety profiles; however, few exhibit pathway-biased mechanisms of action. Here, we report that angelic acid (AA), a naturally occurring unsaturated monocarboxylic acid, potently inhibits RANKL-induced osteoclastogenesis. This effect occurs with an IC50 of 1.9 µM without cytotoxicity. Mechanistically, AA selectively suppressed RANKL-activated phosphorylation of ERK1/2, p38, and JNK (all three MAPK branches), while leaving NF-κB transcriptional activity unaffected. This preferential MAPK suppression disrupted downstream NFATc1 nuclear translocation, thereby preventing NFATc1-driven transcription of osteoclast-specific effector genes including TRAP, cathepsin K, and Atp6v0d2. These findings identify AA as a novel inhibitor of the RANKL-MAPK-NFATc1 axis, providing a mechanistic foundation for its therapeutic development in osteoporosis and other osteolytic diseases.

Alzheimer's disease (AD) is a multifactorial neurodegenerative disorder characterized by cholinergic dysfunction, amyloid-β aggregation, mitochondrial stress, and aberrant kinase activity. Carotenoids, naturally occurring pigments with antioxidant and neuroprotective properties, have emerged as promising candidates for AD intervention. In this study, we performed a systematic stepwise computational screening of a large carotenoid library (n = 1191) to identify multitarget candidates against AD-related proteins. The workflow consisted of predefined ADMET filtering (oral absorption > 90%, Caco-2 > 0.9, logBB > -1, and absence of major CYP inhibition and toxicity alerts), reducing the dataset to 61 compounds, followed by multi-target molecular docking against AChE, BChE, BACE-1, MAO-B, and GSK3-β. Compounds were ranked using an aggregated mean docking score across all five targets, and the top-performing candidate was subjected to detailed mechanistic analyses. Hopkinsiaxanthin emerged as the highest-ranked multitarget carotenoid and was further evaluated using frontier molecular orbital (FMO) analysis, pharmacophore modeling, 100 ns molecular dynamics (MD) simulations, MM/PBSA binding free energy calculations, and per-residue decomposition. Docking predicted favorable estimated binding affinities toward all targets. MD simulations confirmed stable receptor-ligand complexes with low RMSD values (0.278-0.285 nm). MM/PBSA analysis indicated favorable binding free energies, particularly for GSK3-β (-22.73 kcal/mol) and AChE (-21.50 kcal/mol). Per-residue decomposition identified key hotspot residues driving stabilization. Overall, this structured computational framework identifies Hopkinsiaxanthin as a promising multitarget scaffold and supports its prioritization for experimental validation in AD models.

Salmonella Enteritidis is a major threat to poultry health and food safety, underscoring the need for safe alternatives to conventional antibiotics. In this study, quercetin, a natural flavonoid with antibacterial and immunomodulatory properties, was evaluated using an integrated approach combining network pharmacology, molecular docking, in vitro antibacterial assays, and preliminary in vivo validation. Potential targets of quercetin and Salmonella Enteritidis were identified from the TCMSP and GeneCards databases, followed by protein-protein interaction analysis, topological screening, and GO/KEGG enrichment analyses. Five core targets, namely IL1B, IL6, STAT1, PTGS2, and IFNG, were identified and were mainly enriched in immune- and inflammation-related pathways. Molecular docking suggested favorable interactions between quercetin and these predicted targets. In vitro, quercetin showed moderate antibacterial activity against Salmonella Enteritidis, with a minimum inhibitory concentration of 256 μg/mL and a minimum bactericidal concentration of 512 μg/mL. In vivo, quercetin alleviated intestinal histopathological damage and reduced the transcriptional expression of the five target genes in infected chicks in a dose-dependent manner, with more evident effects at doses of 512 mg/kg or higher. These findings provide preliminary evidence that quercetin may exert both direct antibacterial and host-associated protective effects against Salmonella Enteritidis, although the underlying mechanisms require further validation.

Endometrial cancer (EC) is the most common gynecologic malignancy in developed countries, with increasing incidence linked to obesity and metabolic dysfunction. While early-stage EC is often curable, advanced and recurrent disease remains difficult to treat due to resistance and limited therapeutic options. Natural products derived from traditional Chinese medicine have attracted attention as complementary strategies in EC management. These compounds exhibit multi-target effects, including modulation of estrogen signaling, inhibition of proliferation, induction of apoptosis, and regulation of immune and inflammatory pathways. This review summarizes current evidence on natural products in EC, integrating preclinical findings, emerging clinical data, and mechanistic insights from molecular and systems biology approaches. Key challenges, including variability, bioavailability, and insufficient clinical validation, are discussed. Future directions emphasize the integration of natural products into precision oncology frameworks.

Attention-Deficit/Hyperactivity Disorder (ADHD), affecting 5-10% of children globally, faces treatment limitations due to adverse effects and uncertain long-term risks of current pharmacotherapies. This study investigated the therapeutic potential of sepia ink (SI), a marine-derived natural complex from cuttlefish, in a scopolamine-induced ADHD-like mouse model. The chemical constituents of SI were characterized via Ultra-Performance Liquid Chromatography-Mass Spectrometry (UPLC-MS). The behavioral assessments, histopathological examinations, flow cytometry, and complete blood counts were utilized to evaluate its effects on ADHD-like phenotypes, neuroinflammation, and immune function. Integrated transcriptomic, plasma metabolomic, and 16S rRNA sequencing were used to explore the underlying mechanisms. SI significantly alleviated hyperactivity and improved spatial learning and memory deficits. It reduced hippocampal neuronal damage, attenuated neuroinflammation, and reversed scopolamine-induced immunosuppression in spleen and thymus. SI also restored the balance of immune cell subsets in both mesenteric lymph nodes and spleen, and the peripheral blood cell counts. Multi-omics analyses suggested that the beneficial effects of SI were associated with reduced neuroinflammation, rebalanced systemic immune responses, partial correction of lipid metabolic disturbances, and restoration of gut microbiota homeostasis. Collectively, our findings indicate that SI effectively mitigates the in vivo ADHD-like impairments by coordinating immune, metabolic, and gut microbiota-related processes, thereby supporting its potential as a marine-derived therapeutic candidate for further ADHD treatment.

We utilized molecular dynamics (MD) simulations to explore the interaction of the RGD peptide with the αvβ3 integrin receptor, a key process for targeted drug delivery to tumors. The goal of these simulations was to model the transport of boron atoms by the RGD peptide and to characterize the binding event between this vector and its receptor. The study focused on the interaction processes and spatial arrangements of the solvated RGD-integrin system. Simulations were run for 100 ns to achieve relaxed-state configurations. Our model featured two RGD peptides: one pre-localized within the integrin's binding site and another initially positioned externally. The external peptide was observed to diffuse freely and subsequently bind to the αvβ3 integrin. This spontaneous binding event provides valuable insights into the pharmacological specificity and mechanisms of the RGD-integrin interaction, informing the design of effective drug delivery systems.

Obesity represents one of the most critical global public health challenges. Pancreatic lipase (PL) serves as a key therapeutic target for obesity control, whereas clinical synthetic PL inhibitors are greatly restricted by adverse reactions. Traditional Chinese medicines (TCMs) have a long-standing history in regulating lipid metabolism and ameliorating obesity-related disorders, and are characterized by remarkable structural diversity, low toxicity, and mild side effects, thus representing a promising source for developing safe and efficient PL inhibitors. In this work, an integrated strategy combining in silico screening and in vitro validation was employed to identify potential PL inhibitors from TCM components, including molecular docking, molecular dynamics simulation, MM/PBSA binding free energy computation, and in vitro enzymatic assay. Six compounds with docking scores ranging from -9.9 to -9.0 kcal/mol were selected for further investigation. Molecular dynamics simulations verified the favorable structural stability of the corresponding ligand-PL complexes, and MM/PBSA calculations demonstrated negative binding free energies from -21.24 ± 0.39 to -12.03 ± 0.40 kcal/mol. In vitro experiments indicated that three compounds (Hydroxygenkwanin, Atractylenolide I, and Peiminine) showed effective PL inhibitory activity, with IC50 values of 0.128 ± 0.009, 0.584 ± 0.031, and 0.748 ± 0.042 mM, respectively. These values are comparable to quercetin (0.231 ± 0.034 mM) but significantly higher than orlistat (0.481 ± 0.023 μM), which is attributed to their non-covalent binding pattern. Collectively, this study validated the reliability of the integrated in silico and in vitro screening strategy, identified three effective pancreatic lipase inhibitors derived from TCMs, established a robust paradigm for the discovery of natural PL inhibitors, and laid a solid foundation for subsequent research on natural anti-obesity agents.

Early identification of ischemic heart failure (IHF) is critical for improving patient prognosis and clinical outcomes. However, effective diagnostic biomarkers and targeted therapeutic strategies for IHF remain limited. Total flavonoids from Dracocephalum moldavica L. (TFDM) exert potential cardioprotective effects; however, the molecular mechanisms by which TFDM acts against IHF have not been fully elucidated. Therefore, this study aims to identify diagnostic biomarkers for IHF and explore the potential therapeutic mechanism of TFDM targeting these key genes. Given the small sample size (n = 17) of the clinical dataset, LASSO regression and Random Forest were employed due to their superior performance in feature selection, noise reduction, and stability in small-sample scenarios. In this study, we screened key characteristic genes of IHF through bioinformatics analysis and further investigated the binding potential between these key genes and active components of TFDM using molecular docking, thus providing new targets for the early diagnosis of IHF and new evidence for the intervention mechanism of TFDM in IHF.