Melanoma represents a formidable challenge in dermatological oncology due to its resistance to conventional treatments. The Celandine Alkali Injection Formula (CAIF) offers benefits on clinical internal medicine treatments, within which chelidonine and tetrandrine are recognized as potential quality markers. However, their synergistic mechanisms facilitating their anti-melanoma action remain unveiled. This study embarked on an exploration of CAIF’s therapeutic potential through a multifaceted research design, integrating system pharmacological predictions with empirical molecular biological evaluations. The dual application of chelidonine and tetrandrine within CAIF exhibited a pronounced inhibitory effect on the proliferation of B16F10 cells, surpassing the effectiveness of individual compound administration. Computational predictions identified the top 50 targets, involved in key signaling pathways including cell cycle regulation, and melanogenesis. RNA sequencing further elucidated that the combinatory treatment modulated a broader spectrum of differentially expressed genes, implicating crucial biological processes including cell differentiation, and tyrosinase metabolism. The combination markedly enhanced melanogenesis and apoptotic indices, arrested cell cycle progression, and fostered cellular differentiation. Notably, chelidonine additionally curtailed the migratory capacity of B16F10 cells. Our findings underscore the therapeutic potential of chelidonine and tetrandrine, key components of CAIF, in effectively combating melanoma by targeting cell proliferation, migration, differentiation, and melanogenesis.

This research centers on the sand-fixing plant known as Stipagrostis pennata, from which the β-glucosidase gene SpBGLU25 was successfully cloned using the molecular cloning method. SpBGLU25 encodes a hydrophilic and stable protein made up of 193 amino acids, located in the cell membrane. qRT-PCR analysis indicated that the expression of the SpBGLU25 is closely linked to the drought stress tolerance of S. pennata. Following this, functional validation was performed using an Arabidopsis overexpression system. The overexpression of transgenic Arabidopsis lines showed significantly improved drought tolerance under PEG and mannitol treatments. Assessments of germination, root length, and physiological indicators such as proline, malondialdehyde content, soluble sugars, and relative leaf water content (RLWC) further confirmed the enhanced performance of the overexpressing plants. Additionally, the comparative transcriptomic analysis of SpBGLU25-OE Arabidopsis compared to the wild-type (WT) showed that differentially upregulated genes were primarily enriched in categories of “cellular process,” “cell,” and “catalytic activity.” KEGG pathway enrichment analysis indicated that the genes were mainly concentrated in the pathways of phenylpropanoid biosynthesis and plant hormone signal transduction. These findings provide a crucial foundation for further investigation into the function of the SpBGLU25 and its role in regulating plant tissue development and adaptation to stress. This research is anticipated to offer new theoretical insights and genetic resources for enhancing plant stress tolerance through genetic engineering.

Atherosclerosis (AS) is emerging as a major global public health problem. Researchers are developing and implementing various anti-AS strategies. This study aimed to investigate gut microbiota and metabolite changes associated with elevated serum low-density lipoprotein cholesterol (LDL-c). Mice were divided into two equal groups: Group C (mice with gut microbiota from healthy subjects) and Group B (mice with gut microbiota from AS). At the end of the 8-week study, blood samples were collected for blood lipid analysis; rectal feces were collected for microbial 16S rRNA sequencing analysis and metabolomics analysis. Our results showed that the B group significantly increased serum lipid levels of LDL-c. However, no statistically significant differences were observed in the richness and diversity of the gut microbiota, but we observed an increase in the ratio of Firmicutes to Bacteroidetes and an increase in the abundance of Parabacteroides Goldstein. In addition, untargeted metabolomic analysis of fecal samples revealed 128 metabolites that were differentially expressed between groups C and B. Notably, group B was found to have significantly increased levels of metabolites involved in lipid metabolism pathways, such as estrogen glucuronide, ginsenoside f1, Pe (16:1e/14,15-epete), and prostaglandin E1. Those data highlight the importance of understanding AS from the gut microbiota perspective and establish a foundation for future research on AS.

Allene oxide cyclase (AOC) is an integral enzyme in jasmonic acid (JA) biosynthesis and a key regulator of plant stress responses. However, the regulatory mechanisms and enzymatic activities of cotton AOC under stress responses remain poorly understood. In our research, using BLASTP and conserved domain validation, we identified 68 AOC gene members across 10 Gossypium species. Phylogenetic, gene-structure, and syntenic analyses showed high evolutionary conservation of AOC genes, accompanied by lineage-specific expansion. Promoter sequence analysis indicated that the AOCs contained abundant cis-elements associated with hormone signaling, abiotic-stress response, and transcription factor (TF) regulation. Expression profiling revealed tissue-specific and stress-responsive expression patterns among GhAOC genes, with GhAOC3A showing the most pronounced upregulation under chilling stress. GhAOC3A was identified as a chloroplast-localized protein based on the subcellular localization assay. GhAOC3A overexpression enhanced cotton chilling tolerance with elevated JA accumulation, stronger antioxidant defense, stable osmotic homeostasis, and retained chlorophyll content, and VIGS results provided supportive evidence for the function of GhAOC3A in chilling tolerance. Among the predicted TFs that could regulate the expression of GhAOC3A, GhWRKY65 was further identified and confirmed to bind to the W-box element and activate GhAOC3A transcription. Overexpression of GhWRKY65 significantly enhanced cotton chilling tolerance, as well as elevated GhAOC3A expression and increased JA accumulation. Conversely, silencing GhWRKY65 compromised cotton chilling tolerance. Collectively, these findings suggest that the GhWRKY65-GhAOC3A regulatory module positively modulates cotton chilling tolerance and JA levels. Our study provides insights into the transcriptional network associated with plant stress response and offers effective candidates for breeding chilling-tolerant cotton materials.

Verticillium wilt (VW), caused by the soil-borne fungus Verticillium dahliae, is a major disease that markedly compromises both the yield and fiber quality of cotton. In this study, we explored the function and underlying mechanism of the cotton expansin gene GhEXLB2 in response to VW infection. Expression profiling revealed that members of the GhEXL family exhibit distinct patterns across tissues and under various biotic and abiotic stresses. Notably, GhEXLB2, which encodes an extracellular protein, showed the strongest induction following V. dahliae challenge. Ectopic expression of GhEXLB2 in Arabidopsis thaliana promoted root elongation and root hair formation, and was associated with improved resistance to the pathogen. In contrast, silencing GhEXLB2 in cotton via virus-induced gene silencing (VIGS) led to pronounced vascular browning, increased pathogen recovery, and a lower level of disease resistance. In addition, RNA-seq profiling of GhEXLB2-silenced (VIGS) cotton plants revealed that most differentially expressed genes were enriched in pathways related to phytohormone signaling and plant–pathogen interactions, with salicylic acid (SA) signaling and WRKY transcription factors emerging as central regulatory components. Analysis of the GhEXLB2 promoter further identified multiple cis-acting elements associated with stress and hormone responsiveness. When integrated with protein–protein interaction (PPI) prediction data, these results suggest that GhEXLB2 may be modulated by a network of transcription factors and signaling pathways. Collectively, the evidence supports a positive association between GhEXLB2 and VW resistance. This study provides a framework for understanding expansin functions in cotton defense against VW.

Metabolic reprogramming is closely linked to tumor proliferation, invasion, and immune escape. Despite its central role in amino acid metabolism, the regulatory mechanisms of asparagine metabolism in hepatocellular carcinoma (HCC) progression remain poorly characterized. Rather than focusing on canonical metabolic genes, prognostic markers were identified from co-expression modules associated with asparagine metabolism signatures. Using the TCGA database and asparagine metabolism-related gene sets, a prognostic risk-scoring model was developed through differential expression analysis, univariate Cox regression, and the LASSO algorithm and externally validated with the GEO dataset (GSE14620). Survival analysis, ROC curve evaluation, nomogram construction, scRNA-seq, GSEA, and drug sensitivity analysis were performed to systematically delineate the molecular mechanisms by which asparagine metabolism drives HCC progression. A three-gene signature comprising BOP1, SAC3D1, and PDE2A effectively stratified patients into high- and low-risk groups. High-risk patients exhibited markedly poorer overall survival, enrichment in tumor proliferation-associated pathways, increased tumor purity, reduced immune cell infiltration, and a substantially higher TP53 mutation rate (38% vs. 13%). In contrast, the low-risk group showed enrichment in pathways linked to hepatoblastoma suppression and liver function, alongside improved predicted response to immunotherapy. Single-cell analysis identified NK cells and endothelial cells as central mediators of asparagine metabolism-driven HCC progression, with BOP1, SAC3D1, and PDE2A displaying dynamic expression patterns during differentiation. Furthermore, the high-risk group was predicted to be more sensitive to chemotherapeutics such as cyclophosphamide and 5-fluorouracil. These findings highlight a potential interplay between nitrogen metabolism and asparagine metabolism in HCC and suggest mechanisms by which these pathways may influence NK cell and endothelial cell function to promote disease progression. This study establishes a novel prognostic model and identifies potential chemotherapeutic vulnerabilities in high-risk patients, warranting further experimental and clinical validation.