Protein kinase D1 (PRKD1), a serine/threonine kinase of the PKD family, has been implicated in tumor biology, but its role in hepatocellular carcinoma (HCC) remains unclear. We explored PRKD1 expression and function using immunohistochemistry, bulk and single-cell RNA sequencing, and in vitro functional assays. PRKD1 protein levels were significantly elevated in 339 HCC tissues compared to corresponding adjacent non-tumorous samples (11.390 ± 1.560 vs. 6.277 ± 2.357, P < 0.0001). Multicenter bulk transcriptomic data confirmed consistent PRKD1 mRNA overexpression (SMD = 0.26, 95% CI = 0.14-0.39), with single-cell transcriptomic profiling indicating specific enrichment in endothelial cells, smooth muscle cells, and hepatocytes. High PRKD1 expression was associated with advanced tumor stages and worse overall survival. Functionally, PRKD1-associated genes were enriched in extracellular matrix and focal adhesion pathways. Immune profiling revealed positive correlations with M2 macrophages, regulatory T cells (Tregs), and myeloid-derived suppressor cells (MDSCs), and negative correlations with CD8+ T cells and CD4+ Th1 cells, suggesting an immunosuppressive role. Mechanistic experiments demonstrated that conditioned medium from PRKD1-knockdown HCC cells promoted M1 polarization and reduced M2 markers in THP-1 cells, while PRKD1 silencing increased PD-L1 and IDO1 expression. In contrast, IFN-γ treatment did not induce PRKD1 expression, indicating that PRKD1 actively contributes to, rather than responds to, immunosuppressive cues. PRKD1 knockdown markedly impaired HCC cell proliferation and migration. Pharmacologically, nitidine chloride significantly reduced PRKD1 expression in a dose-dependent manner, and molecular docking suggested a potential direct interaction. With respect to drug response, PRKD1-high HCC cases exhibited increased predicted sensitivity to multiple tyrosine kinase inhibitors (TKIs), while in vitro PRKD1 knockdown reduced sorafenib sensitivity, and sorafenib treatment suppressed both PRKD1 and p-ERK1/2 levels. Collectively, our findings identify PRKD1 as a multifaceted contributor to HCC progression, immune microenvironment modulation, and TKI responsiveness. These results highlight PRKD1 as a promising therapeutic target warranting further mechanistic and translational investigation.

Heat shock factor (HSF) family proteins modulate ferroptosis in various tumors. We previously confirmed that HSF4 performs a carcinogenic role in colorectal cancer (CRC), but its function in ferroptosis remains unclear. Therefore, this study aims to reveal whether HSF4 regulates the ferroptosis process in CRC and its potential molecular mechanisms. This study found that HSF4 overexpression markedly attenuated Erastin-induced cell death and mitochondrial damage in HT29 and HCT116 cells. HSF4 effectively reduced lipid peroxidation and Fe2+ levels in CRC cells and reversed Erastin-induced changes in ferroptosis marker, including GPX4, SLC7A11, and ACSL4. ChIP-seq combined with GEO dataset analysis identified 249 potential HSF4 target genes, among which the promoter region of the lipid metabolism-related gene MBOAT1/2 binds to HSF4 and is transcriptionally activated. In vitro experiments, MBOAT1/2 knockdown reversed the inhibitory effect of HSF4 overexpression on CRC cell death, mitochondrial damage, lipid peroxidation, Fe2 + accumulation, and changes in ferroptosis marker. In vivo experiments, MBOAT1/2 knockdown effectively reduced tumor volume and downregulated the number of Ki-67-positive cells, GPX4, and SLC7A11, while upregulating ACSL4. In conclusion, HSF4 alleviates ferroptosis in CRC cells and facilitates tumor progression by upregulating MBOAT1/2 transcription, thereby limiting lipid peroxidation and Fe2+ accumulation.

Fungiculture refers to the deliberate cultivation or agricultural practice involving the growth and management of fungi. The practice encompasses the intentional culture of diverse species of macrofungi, including mushrooms and truffles, within controlled habitats or under specified conditions, in order to fulfill human requirements especially for food purpose. As the global market for edible mushrooms grows quickly, it is becoming increasingly necessary to grow novel and improved strains of edible fungi. Growing and breeding edible fungi using traditional methods is both time-consuming and difficult. So, there is a need for evolving advanced techniques at a molecular level which can help breeding of edible fungi with much better efficiency. The CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease 9) system is one of the most effective techniques for accurately cutting and modifying the genomes of edible fungi. In this review, we discuss how genome editing using CRISPR/Cas9 has been utilized in many edible fungal species such as Pleurotus ostreatus, Agaricus bisporus, Cordyceps militaris, Ganoderma lucidum, Flammulina filiformis, Lentinula edodes, and others for their target specific breeding. We also discuss the working mechanism of the above-mentioned system in these mushroom species, and also the advantages and limitations of using this system in mushrooms.

Metabolic-associated fatty liver disease (MAFLD) is a globally prevalent liver disorder, and long non-coding RNAs (lncRNAs) play a crucial role in its pathogenesis. However, the specific function of taurine up-regulated gene 1 (TUG1) remains incompletely understood. This study examined the molecular interactions among TUG1, miR-29a-3p, 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR) and sterol regulatory element-binding protein 2 (SREBP-2). Two MAFLD mouse models were established, and liver damage and lipid accumulation were assessed using hematoxylin and eosin (HE) and Oil Red O staining. The expression levels of these factors were measured in liver tissues, and TUG1 expression was knocked down to evaluate its functional impact. The results indicate that TUG1 could bind to miR-29a-3p, thereby regulating the expression of HMGCR and SREBP-2. Tug1 expression is upregulated in livers of MAFLD models compared with control groups, showing a negative correlation with miR-29a-3p and a positive correlation with HMGCR and SREBP-2. Furthermore, the knockdown of Tug1 in mouse livers reduced hepatic lipid deposition significantly. In summary, TUG1 could modulate the SREBP-2/HMGCR pathway by binding to miR-29a-3p, thus influencing lipid metabolism. These findings suggest that TUG1 may serve as a potential therapeutic target for MAFLD.

Breast cancer-related depression (BCRD) is a prevalent comorbidity that markedly reduces quality of life and can negatively influence treatment outcomes. The molecular basis of BCRD remains elusive, particularly the role of the IL-17 signaling pathway in the interaction between breast cancer and depression.We established a BCRD mouse model by inducing breast tumors and administering chronic corticosterone. Transcriptomic analysis was performed on brain and tumor tissues to identify differentially expressed genes (DEGs) associated with BCRD. Functional enrichment analyses were conducted to determine the biological functions and signaling pathways linked to these DEGs. For in vitro validation, lipopolysaccharide (LPS)-stimulated BV2 microglia cells were used to mimic neuroinflammation, and the effects of modulating IL-17 signaling on cellular activation were assessed.In vivo, BCRD mice exhibited increased immobility time in the tail suspension test and reduced sucrose preference, indicative of depressive-like behaviors. Transcriptomic analysis revealed substantial changes in immune-related genes, particularly those involved in the IL-17 signaling pathway. In vitro, LPS stimulation elevated IL-17, NF-κB p65, and other inflammatory markers in BV2 cells, whereas IL-17 inhibition attenuated these responses. Furthermore, we observed increased expression of IL-17 and NF-κB p65 in the brain tissues of BCRD mice, which was associated with increased microglial activation and blood-brain barrier permeability.These findings demonstrate that the IL-17 signaling pathway plays a crucial role in BCRD development, linking breast cancer progression to depressive symptoms through microglia activation and disruption of the blood-brain barrier. Targeting the IL-17 signaling pathway may offer a promising therapeutic strategy for treating BCRD.

Glycine C-acetyltransferase (GCSH), a component of the mitochondrial glycine cleavage system, has been previously linked to cuproptosis or tumor progression, but its role in colorectal cancer (CRC) remains incompletely understood. We analyzed GCSH expression across multiple CRC datasets, including the TCGA dataset (n = 689) and four independent GEO datasets (total n = 709), and validated findings in clinical tissues, blood samples, and CRC cell lines. Functional roles of GCSH were assessed using CCK-8, wound healing, and Transwell assays. Cuproptosis was induce, and evaluated by flow cytometry, Western blotting, and measurement of intracellular Cu2+, ROS, and FDX1 levels. The regulatory relationship between GCSH and FDX1, as well as the involvement of the PI3K/AKT pathway, was explored through rescue experiments and molecular docking analysis. The analysis of Bulk RNA-seq data and scRNA-seq datasets revealed the expression and the biological function of GCSH in CRC. The validation experiments confirmed that GCSH was consistently overexpressed in CRC tissues and blood samples, and its expression correlated with distant metastasis. GCSH knockdown inhibited cell viability, migration, and invasion, with minimal effects on apoptosis. Mechanistically, GCSH suppressed cuproptosis by downregulating FDX1 protein and reducing intracellular Cu2+ and ROS accumulation. Molecular docking suggested a potential interaction between GCSH and FDX1. Furthermore, GCSH was identified as a downstream effector of PI3K/AKT pathway, mediating its protective effect against cuproptosis. GCSH promotes CRC progression and confers resistance to cuproptosis via the PI3K/AKT-GCSH/FDX1 axis. These findings identify GCSH as a novel regulator of cuproptosis and a potential therapeutic target in CRC.

Feather follicles are specialized skin appendages that are essential for thermoregulation, protection, and down production in birds, forming through complex genetic and epigenetic interactions during embryogenesis. In this study, we examined skin and follicle development in Hungarian white goose embryos, focusing on dynamic epigenetic-transcriptomic changes. Histology showed smooth epidermis at E10, feather buds at E13, and columnar follicles with medullary tissue and secondary follicles at E18. Transcriptomics revealed 1327 and 1847 DEGs enriched in epidermal development, differentiation, and adhesion. Primordial initiation at E10-E13 featured Wnt, TGF-β, and melanogenesis, whereas follicle formation at E13-E18 involved lipid metabolism and VEGF signaling. Keratinization genes were continuously upregulated, and Wnt, Shh, and muscle pathways were activated late. Key regulators included LEF1, MSX2, and FOXN1. ATAC-seq showed dynamic chromatin accessibility, stage-specific promoter openness, and motifs for YY1, KLF5, and KLF4. Differentially accessible regions enriched genes in Wnt and TGF-β signaling, cell adhesion, and mitophagy, shifting from proliferation and basic metabolism at E10-E13 to differentiation, lipid metabolism, and homeostasis at E13-E18. Integrated analyses linked fatty acid metabolism, MAPK, and FoxO signaling to differentiation, while downregulation of redox and migration pathways preserved homeostasis. Some fatty acid metabolism and cell polarity genes increased expression despite reduced accessibility at E13-E18, indicating epigenetic pre-programming and post-transcriptional interplay. This work delineates coordinated epigenetic-transcriptional regulation of goose embryonic skin and feather follicle morphogenesis, offering insights for avian and vertebrate skin appendage studies.

The global rise of antimicrobial resistance has intensified the demand for novel antimicrobial agents with broad-spectrum efficacy and unique mechanisms of action. Herein, a marine-derived strain, Bacillus velezensis (B. velezensis) AM12, exhibiting clear inhibitory activity against eight major foodborne pathogens, was isolated from coastal seawater near Zhanjiang, China. Whole-genome sequencing revealed a 3,992,266bp circular chromosome with a GC content of 46%, and functional annotation indicated extensive metabolic potential. Genome mining using the antiSMASH platform identified 14 secondary metabolite biosynthetic gene clusters, including those encoding surfactin, macrolactin H, bacillaene, fengycin, difficidin, bacillibactin, and bacilysin, as well as several additional clusters potentially responsible for the biosynthesis of novel antimicrobial compounds. Analysis using the BAGEL4 database further detected four bacteriocin/RiPP biosynthetic loci, corresponding to Competence/ComX, Amylocyclicin, LCI, and a Lichenicidin-like cluster. These collectively explain the broad-spectrum antibacterial activity of AM12 and suggest potential antifungal capability. Notably, the core genes of ComX and LCI exhibited relatively low similarity (46.67% and 73.91%, respectively) to known references, and B. velezensis AM12-origin lichenicidin component 2 showed no detectable homology, suggesting that AM12 may encode structurally and functionally distinct novel bacteriocins. Comparative genomic analysis with closely related terrestrial B. velezensis strains revealed that AM12-specific genes are predominantly enriched in functions associated with marine environmental adaptation and antimicrobial activity. Furthermore, machine learning-based prediction using the AM12-specific gene set identified seven high-confidence antimicrobial peptide candidates characterized by cationic charge, amphipathicity, and structural stability. This study elucidates the genomic and molecular basis of the broad-spectrum antibacterial activity pertaining to B. velezensis AM12, and establishes a systematic framework for the identification of novel antimicrobial peptides, offering candidate genes for subsequent functional characterization.