Colorectal cancer (CRC) remains one of the leading causes of cancer‑related mortality worldwide. Despite advances in targeted therapies, drug resistance and limited efficacy in KRAS‑mutant CRC continue to present clinical challenges. Antrodia cinnamomea, a medicinal fungus, demonstrates antitumor properties; however, the mechanisms of its triterpenoid compound, 4‑acetylantrocamol LT3 (LT4), remain unclear. The present study investigated the effects of LT4 on KRAS‑mutant HCT116 CRC cells using cell viability, colony formation and migration assays. Western blotting was also employed to examine key signaling pathways. Transcriptome profiling via RNA sequencing was followed by Kyoto Encyclopedia of Genes and Genomes/Gene Ontology enrichment and protein‑protein interaction network analyses using STRING, CytoHubba and Molecular Complex Detection (MCODE). Molecular docking with PI3Kγ (PDB: 1E7U) was conducted to evaluate the predicted binding position and docking energy of LT4. The results indicated that LT4 significantly inhibited HCT116 cell proliferation and migration, induced a mesenchymal‑to‑epithelial transition, suppressed PI3K/AKT/mTOR and ERK signaling and activated the GSK3β/FOXO and phosphorylated‑p38/p21 axes. LT4 also reduced the levels of cyclooxygenase‑2 and anti‑apoptotic proteins (Bcl‑2 and Bcl‑XL) and reduced the expression of the mitochondrial respiratory chain protein cytochrome c oxidase subunit IV. Transcriptomic analysis identified the PI3K/AKT pathway as the most significantly enriched signaling cascade. Network topology analyses highlighted solute carrier family 3 member 2, Cyclin D1, phosphoserine aminotransferase 1 and ChaC glutathione‑specific γ‑glutamylcyclotransferase 1 as central nodes, linking the effects of LT4 to nutrient signaling, redox homeostasis and serine metabolism. Molecular docking confirmed that LT4 stably occupied the ATP‑binding pocket of PI3Kγ with a binding energy comparable to wortmannin and a conformation similar to antroquinonol. In conclusion, to the best of our knowledge, the present study is the first to comprehensively demonstrate the multi‑target anti‑CRC effects of LT4, highlighting its potential as a therapeutic agent, especially in KRAS‑mutant CRC.

MAGE family member A1 (MAGEA1), a cancer‑testis antigen (CTA), is aberrantly expressed in several malignancies such as lung and liver cancers. However, its role in cervical cancer remains to be elucidated. The present study investigated the functional significance and therapeutic potential of MAGEA1 in cervical cancer using lentiviral short hairpin RNA‑mediated knockdown, a series of functional assays, RNA sequencing (RNA‑seq), and nude mouse xenograft models. It was found that MAGEA1 was upregulated in cervical cancer cells and its knockdown substantially suppressed cell proliferation, migration, invasion, and in vivo tumor growth. RNA‑seq analysis further revealed that MAGEA1 silencing altered pathways related to apoptosis, DNA repair, and metabolism. Moreover, MAGEA1 knockdown enhanced chemosensitivity, indicating a potential role in mediating drug resistance. Collectively, the findings identified MAGEA1 as a key oncogenic driver in cervical cancer and highlighted its promise as both a prognostic biomarker and a therapeutic target, offering novel avenues for personalized treatment strategies in cervical cancer.

The global rise of antimicrobial resistance (AMR) in Enterobacterales, including the Enterobacter cloacae complex, is narrowing treatment options. Tigecycline, a last-resort antibiotic for the treatment of multidrug-resistant (MDR) Gram-negative pathogens, is increasingly compromised by emerging resistance mechanisms, notably efflux pump overexpression and regulatory network adaptation. In this study, sixty clinical isolates of Enterobacter cloacae (thirty-eight tigecycline-resistant [TGC-R], twenty-two tigecycline-susceptible [TGC-S]) were analyzed to investigate gene expression changes in efflux pumps and regulatory genes under tigecycline pressure (1/2 minimum inhibitory concentration [MIC]) and standard conditions. Tigecycline exposure markedly increased tolC and acrA together with the regulators marA and ramA, while acrB increased only modestly. This indicates a strong regulatory component to the tigecycline response. In contrast, TGC-S isolates exhibited significant induction of marA, marB without corresponding activation of efflux pumps. Δlog2FC analysis highlighted distinct transcriptional shifts between exposed and unexposed groups, with resistant strains displaying greater divergence. Heatmaps and boxplot visualizations, supported by Wilcoxon test statistics, underscored the regulatory responses associated with tigecycline pressure. These findings indicate that, alongside AcrAB-TolC upregulation, stress-responsive regulators (marA, ramA) are strongly induced by sub-inhibitory tigecycline, underscoring the multifactorial regulation of tigecycline response in the E. cloacae complex.

In Northern Europe, climate warming is driving the northward expansion of deciduous tree species such as aspen and silver birch, while simultaneously intensifying biotic stress from pests and pathogens. This creates an urgent need for improved understanding of molecular defence mechanisms underlying stress resistance and resilience in temperate forest trees, as a basis for the development of innovative biotechnological approaches. However, progress in this area remains limited by the lack of reproducible experimental systems and well-characterized molecular markers for systemic defence responses in deciduous tree species. In this study, we aimed to identify and validate known plant defence gene markers associated with systemic stress responses in hybrid aspen and silver birch to support future functional research. Using sequence mining and phylogenetic analyses, we identified homologues of biotic stress-response genes in the genomes of both species. We then employed in vitro propagated tree clones to assess defence gene activation in distal leaves following systemic signal induction by leaf wounding and bacterial flagellin treatment at 4 and 24 hours post-induction. We identified LOX2, MPK3, and EIN2 as early wounding-responsive genes in silver birch, while JAZ10 together with EIN2 showed robust induction in hybrid aspen in response to the combined effects of wounding and flagellin. Collectively, these findings establish a reproducible in vitro framework for validating stress responsive genes and provide a foundation for future studies of systemic signalling, tree-microbe interactions, and stress resilience in ecologically and economically important forest tree species.

The dysregulation of N6-methyladenosine (m6A) modification drives progression in non-small cell lung cancer (NSCLC), yet its interplay with traditional medicine-derived therapeutics remains largely unexplored. We propose a novel strategy that integrates m6A-based prognostic subtypes with Pueraria pharmacology to identify prognostic markers and therapeutic targets related to m6A regulators for NSCLC treatment. Multi-omics clustering of 1,661 NSCLC samples identified three distinct m6A modification patterns. Based on these, a robust 19-gene prognostic signature was constructed via Cox regression and validated in the GSE31210 dataset. This risk model significantly correlated with immune infiltration and patient survival. Furthermore, the expression patterns of these genes were validated via single-cell RNA-sequencing (scRNA-seq) and RT-qPCR in NSCLC cell lines. To identify pharmacological interventions, we intersected the m6A prognostic signature with 7,333 NSCLC-related genes and 366 Pueraria targets, revealing IGFBP1 as the core therapeutic nexus. Immunohistochemistry confirmed the expression of IGFBP1 in NSCLC tissues. Molecular docking and 100-ns molecular dynamics (MD) simulations confirmed stable binding of Pueraria compounds to IGFBP1, specifically 7,8,4'-trihydroxyisoflavone (binding energy = -8.3 kcal/mol) and genistein (-7.4 kcal/mol). This study establishes IGFBP1 as a therapeutic nexus connecting m6A-driven NSCLC progression and the anti-tumor effects of Pueraria. Our RNA-modification-guided pharmacology approach advances the integration of traditional medicines into precision oncology.

Carbapenem-resistant Acinetobacter baumannii (CRAB) has appeared as a leading cause of hospital-acquired infections, resulting in high mortality rates and limited treatment options. The development of novel antibacterial agents has lagged behind the rapid spread of antibiotic-resistant bacteria; thus, alternative therapeutic strategies are urgently needed. In this study, we investigated plumbagin, a natural compound derived from Plumbago zeylanica L., for its potential antibacterial and antibiofilm activities against CRAB. MIC and MBC determinations showed that plumbagin significantly inhibited growth and exerted bactericidal activity at low concentrations. Biofilm inhibition concentration and biofilm eradication concentration assays revealed that plumbagin both prevented biofilm formation and eradicated mature biofilms. Consistent with these findings, XTT reduction assays showed a marked decrease in metabolic activity after plumbagin treatment, and confocal laser scanning microscopy with COMSTAT analysis confirmed reduced biofilm biomass and decreased viability of biofilm-embedded cells. Further, quantitative polymerase chain reaction confirmed the downregulation of the carbapenem-resistance gene blaOXA-23 and biofilm-related genes, including bfmR, csuA/B, ompA, and bap. Collectively, these results reveal plumbagin as a therapeutic candidate against CRAB.

Despite advances in therapeutic regimens for managing cancer progression, ovarian cancer (OVC) still depends on platinum-based chemotherapy as its first-line treatment. Acquired resistance is accompanied by abnormal alterations in epigenetic regulation; however, in-depth mechanistic studies on cisplatin-resistant OVC are lacking. Herein, we show that abnormal overexpression of histone lysine demethylase 5B (KDM5B), but not KDM5A, strongly correlates with cisplatin resistance and OVC tumor progression. Genome-wide sequencing data revealed that KDM5B removes H3K4me3 from the promoter of dual-specificity phosphatase 4 (DUSP4), activating the MAPK pathway to increase cisplatin resistance. We also found that KDM5B protein stability is dynamically controlled via the ubiquitin-proteasome system (UPS), which is mediated by ubiquitin-specific protease 7 (USP7), F-box and WD repeat domain-containing 7 (FBXW7), and homeodomain-interacting protein kinase 1 (HIPK1). KDM5B and USP7 depletion effectively resensitizes OVC to cisplatin resistance, whereas DUSP4 silencing results in resistance in vitro and in vivo. Targeting KDM5B and USP7 synergistically represses tumor progression and increases sensitivity to cisplatin. Overall, we propose two new UPS-associated proteins, USP7 and FBXW7, which are responsible for abnormal KDM5B protein regulation, and suggest a novel mechanism to overcome cisplatin resistance in OVC by targeting the KDM5B-DUSP4 axis.

Multiple myeloma (MM) is characterized by kidney deficiency, phlegm, and blood stasis as core findings, specifically in Traditional Chinese Medicine (TCM), and the kidney-tonifying, phlegm-resolving, and blood stasis-removing (KPR) method is a fundamental therapeutic approach for MM in TCM. Western medicine primarily focuses on targeted immunotherapy or chemotherapy for MM treatment, whereas TCM characterizes MM through distinct pathological patterns that directly correspond to immune microenvironment dysregulation. Emerging evidence implicates the PHD finger protein 19 (PHF19)/enhancer of zeste homolog 2 (EZH2)/trimethylated histone H3 at lysine 27 (H3K27me3) epigenetic axis in immune microenvironment dysregulation and MM progression. Notably, TCM "blood stasis" correlates with hypoxia-induced immune gene silencing in MM bone marrow, and KPR (a clinically validated TCM decoction with 16 herbs) acts on this axis via its active components that regulate EZH2 and epigenetic function, merging TCM syndrome differentiation with modern epigenetics. We have designed a randomized controlled trial (RCT) to investigate the mechanism of action and safety of the KPR method in MM.

Pyriproxyfen (PPF) and diflubenzuron (DFB) are widely used pesticides with metabolic toxicity in humans underexplored. White adipose tissue (WAT) is a potential target for endocrine-disrupting chemicals.

Esophageal adenocarcinoma (EAC) is an aggressive malignancy with poor patient outcomes and limited therapeutic options. WEE1, a G2/M checkpoint kinase, is often upregulated in cancers and associated with resistance to therapy. In this study, we identify a previously unrecognized cytoplasmic role of WEE1 in stabilizing the oncogenic transcription factor MYC and promoting drug resistance in EAC. WEE1 was found to be aberrantly overexpressed and mislocalized to the cytoplasm in EAC cell lines and patient samples. WEE1 depletion or inhibition by MK-1775 significantly reduced MYC protein levels and transcriptional activity by promoting proteasome-mediated degradation. Mechanistically, WEE1 inhibition activated GSK3β, leading to phosphorylation of MYC at threonine 58 and subsequent ubiquitin-dependent degradation. In contrast, overexpression of wild-type WEE1, but not a kinase-dead mutant (K328A), increased p-CDC2 (Y15) and MYC protein levels, confirming that WEE1's kinase activity is essential for maintaining MYC stability. WEE1 inhibition also downregulated the MYC-ABCC1 axis, decreased MRP1 expression, and impaired drug efflux. A high-throughput screen of 892 FDA-approved compounds identified the histone deacetylase inhibitor Panobinostat as a potent synergistic partner of MK-1775. Combination treatment induced robust apoptosis and markedly suppressed tumor growth in EAC organoids and patient-derived xenograft models. These findings reveal a novel cytoplasmic function of WEE1 in sustaining MYC stability and chemoresistance. Targeting WEE1 destabilizes MYC and enhances therapeutic response, supporting the combination of MK-1775 and Panobinostat as a promising treatment strategy for EAC.

Aristolochic acid (AA), commonly used in Chinese herbal medicine to treat various diseases, can cause acute kidney injury (AKI). The pregnane X receptor (PXR), a nuclear receptor, is involved in drug metabolism, carcinogenesis, inflammation, apoptosis, oxidative stress and energy metabolism. Here, we demonstrate that PXR plays a protective role in AA-induced AKI. First, PXR expression was dramatically decreased in mice and Boston University mouse proximal tubular (BUMPT) cells treated with AA. Overexpression of PXR in BUMPT cells alleviated apoptosis induced by AA in vitro. The specific agonist of PXR pregnenolone carbonitrile (PCN) relieved AA-induced AKI in mice, while the PXR inhibitor ketoconazole exacerbated the damage caused by AA in mice. Mechanistically, PXR bound to p53 in BUMPT cells and led to the ubiquitination and degradation of p53, thereby downregulating its expression. Taken as a whole, our data demonstrate that PXR may protect against AA-induced AKI by suppressing p53 expression.

Changes to organismal growth induced by environmental stress are orchestrated at the cellular level. These periods of stress may be followed by recovery periods, when plants have the opportunity to return to normal growth conditions. However, the cell cycle mechanisms underlying recovery are poorly understood. We tested the cell cycle regulation in roots during control, stress, and recovery period for salt, osmotic, cold, and heat stresses using Arabidopsis thaliana, Brachypodium distachyon, and Lolium multiflorum. During the salt and cold stress conditions, the cell cycle pauses at gap phase and is released from gap phase during stress recovery, which depends on cell cycle regulators CDKA;1 and ICK1. Cold stress and recovery, which affect cell division only, follow a conserved 'pause and play' mechanism of the cell cycle.

Fibroblasts are crucial in tissue engineering because of their ability to synthesize the extracellular matrix (ECM) and secrete growth factors. Orbital fibroblasts (OFs) from patients with thyroid eye disease (TED) exhibit enhanced fibroblastic properties, making them ideal candidates for regenerative medicine in ocular tissue. In the present study, we investigated the effect of nanoperlite on TED OFs. Nanoperlite, with its unique properties including high silica (SiO2) content, holds promise for enhancing fibroblast functions. Nanoperlite was prepared and characterized in terms of particle size and chemical composition. A sample of orbital adipose tissue was taken from a TED patient during orbital decompression surgery and OFs were expanded in vitro. The cells were then treated with nanoperlite at concentrations of 1 and 10 μg/mL for 24 h, and gene expression related to the fibrogenesis process was assessed using real-time PCR. Nanoperlite at 1 μg/mL significantly increased the expression of TGF-β, CD90, α-SMA, ZEB1, β-Catenin, and Snail genes in OFs. However, at 10 μg/mL, this effect was not observed. This study highlights nanoperlite's potential to enhance fibroblast activity specifically at the concentration of 1 μg/mL. This effect can potentially aid tissue engineering strategy for periorbital tissue repair and eyelid reconstruction. However, further research is needed to fully elucidate its therapeutic potential and safety profile.

Alcoholic liver disease (ALD) is driven by oxidative stress, lipid metabolism, inflammation, and apoptosis. Current therapies lack efficacy in targeting multi-pathway mechanisms. Xing-Pi-Qing-Gan decoction (XPQG) is an improved traditional Chinese medicine designed to alleviate ALD, but its molecular mechanism remains unknown.

The present study investigated the effects of different humic acid (HA) and zinc (Zn) application methods on membrane durability index (MDI), leaf relative water content (RWC), turgor loss (TL), proline content (PC), and C-repeat binding factor (CBF) gene expressions in oat plants exposed to various low temperatures. For this purpose, two oat cultivars-Albatros (cold-sensitive) and Checota (cold tolerant)-were grown under controlled conditions with HA and Zn applied to the seeds or soil, either individually or in combination, until the 3-4 leaf stage. The plants were subsequently exposed to temperatures of 4 °C, 0 °C, -5 °C, -10 °C, and -15 °C for 24 h each. The results indicated that the application methods of HA and Zn substantially influenced the plants' responses to low temperature. Among the treatments, soil application of HA+Zn (SA_HA+Zn), seed priming with HA combined with soil-applied Zn (SP_HA+SA_Zn), and seed priming with Zn combined with soil-applied HA (SP_Zn+SA_HA) provided the greatest protection against cold stress, as evidenced by improved MDI, RWC, TL, and PC levels. Gene expression analyses further revealed that low temperatures upregulated the CBF genes and the related regulatory genes VRN1 and ZAT12, with the strongest induction observed under SA_HA+Zn, suggesting that this combined approach more effectively activates the plant's cold defense mechanisms.

Pyroptosis, a type of programmed cell death, exerts direct influence on inflammatory processes and immune response. A previous study suggests that miltirone exhibits notable anti-tumor activities and has been shown to induce tumor cell pyroptosis. Nevertheless, the therapeutic value of miltirone in kidney renal clear cell carcinoma (KIRC) remains underexplored.

KRASG12C inhibitors, such as sotorasib, show clinical efficacy for non-small cell lung cancer (NSCLC) positive for the G12C mutations of KRAS, but primary and acquired resistance to these drugs remains a clinical problem. In this study, we show that the development of resistance to sotorasib in KRASG12C-positive NSCLC cells was mediated by constitutive activation of EGFR resulting from downregulation of the protein tyrosine phosphatase receptor type R (PTPRR). PTPRR has been identified as a physiologic regulator of ERK signaling in several cancer types. In our study, PTPRR was demonstrated to bind directly to EGFR, facilitating its dephosphorylation on tyrosine residues. Resumption of PTPRR expression in the resistant cells attenuated EGFR phosphorylation and restored sotorasib sensitivity. PTPRR downregulation was associated with gene promoter hypermethylation in the sotorasib-resistant cells and NSCLC tissue samples. Furthermore, low PTPRR expression in tumor specimens was associated with shorter progression-free and overall survival for patients with NSCLC treated with sotorasib. In contrast to sotorasib, high PTPRR expression was associated with a poor response to EGFR tyrosine kinase inhibitors in EGFR-mutated NSCLC, suggesting that PTPRR may broadly regulate EGFR dependence in NSCLC. Finally, dual blockade of KRASG12C and EGFR showed a substantial antitumor effect in a xenograft model of sotorasib-resistant NSCLC. This approach is therefore a rational therapeutic strategy for KRASG12C-positive NSCLC, especially for tumors showing PTPRR downregulation.

Glioblastoma (GBM) is the most aggressive primary brain tumour, associated with a dismal prognosis and an urgent need for innovative therapeutic strategies. To address this challenge, our group developed DMC-GF, a novel brain-targeted curcumin analog engineered to enhance blood-brain barrier permeability by blocking metabolic sites and improving GLUT1 recognition. Although its activity against glioma stem cells has been reported, the direct mechanisms by which DMC-GF acts on GBM cells remain unclear. In this study, we systematically investigated the molecular actions of DMC-GF using phenotypic assays, transcriptome sequencing, and bioinformatics analysis. DMC-GF exerted dose-dependent inhibitory effects on GBM cell proliferation, migration and invasion and concurrently promoted apoptosis, as reflected by reduced Bcl-2 expression, activation of Bax/Caspase-3 and reversal of epithelial-mesenchymal transition (E-cadherin↑, N-cadherin↓, MMP-3↓). Transcriptomic profiling identified THBS1 as a key downstream target, showing marked suppression following DMC-GF treatment. Functional experiments further confirmed that THBS1 knockdown mimics the anti-tumour effects of DMC-GF, whereas THBS1 overexpression partially mitigates its inhibitory actions. Mechanistic studies revealed that DMC-GF suppresses the non-canonical, Smad-independent TGF-β1 pathway by downregulating THBS1, thereby inhibiting PI3K/AKT signalling, as reflected by reduced phosphorylation of AKT, GSK3β and mTOR. Collectively, this work provides the first evidence that DMC-GF exerts anti-GBM effects through modulation of the THBS1/TGF-β1/PI3K-AKT axis. These findings suggest DMC-GF as a compelling brain-targeted therapeutic candidate, providing new mechanistic insights and a potential clinical strategy to overcome therapeutic resistance in GBM.

BRAF inhibitors (BRAFi) have transformed the treatment of BRAF mutant melanoma, but inherent and acquired resistance remains a major barrier to curative outcomes. Resistance arises from interconnected mechanisms: genetic alterations reactivating the MAPK pathway or bypass cascades (e.g., PI3K/AKT/RTK), epigenetic modulation, metabolic reprogramming, and the tumor microenvironment (TME) remodeling. Despite extensive research into these mechanisms, a cohesive framework linking each resistance module to targeted therapeutic strategies is lacking. This review systematically categorizes resistance into intrinsic and acquired subtypes: intrinsic resistance is driven by constitutive molecular traits of BRAF mutant melanoma (e.g., persistent MAPK activation, baseline PI3K/AKT hyperactivity), while acquired resistance emerges via therapeutic pressure-induced genetic mutations, epigenetic shifts, metabolic reprogramming, or TME modifications. For each identified resistance mechanism, we provide a detailed examination of corresponding therapeutic advancements. These encompass the development of next-generation BRAFi, strategically designed combination therapies, epigenetic modulators, immunotherapeutic approaches, and RNA-based therapeutic agents. Furthermore, we underscore the pivotal role of state-of-the-art technologies, such as liquid biopsies, single-cell multi-omics analyses, and artificial intelligence, in facilitating precise resistance monitoring and personalized therapy selection. By integrating these insights, we present a structured, translationally focused framework to guide basic research and clinical decision-making, ultimately advancing precision salvage therapy and trials aimed at preventing or overcoming BRAFi resistance.

Programmed cell death (PCD) is a major cause of reduced cell viability following cryopreservation, yet the underlying mechanism remains unclear. In this study, pollen from Paeonia lactiflora was used as the experimental material to investigate the role of the microtubule cytoskeleton in PCD during pollen cryopreservation, which exhibits significant viability decline after cryopreservation. The results showed that post-cryopreservation addition of the microtubule-depolymerizing agent oryzalin significantly decreased pollen viability. This effect was accompanied by the activation of caspase-like proteases, reduced mitochondrial membrane potential, elevated intracellular cytochrome C levels, accumulation of PCD signaling molecules, and ultimately increased apoptosis rates. In contrast, treatment with the microtubule-stabilizing agent paclitaxel exerted the opposite effect. At the transcriptional level, paclitaxel treatment induced 754 differentially expressed genes (DEGs); oryzalin treatment resulted in 575 DEGs, a total of 63 DEGs were shared between the two treatments. At the protein level, paclitaxel treatment yielded 262 differentially expressed proteins (DEPs), while oryzalin treatment led to 270 DEPs, with 100 DEPs overlapping between the two groups. Integrated transcriptomic and proteomic analyses revealed that these DEGs and DEPs were significantly enriched in two key pathways: cysteine and methionine metabolism, and protein processing in the endoplasmic reticulum. Notably, heat shock proteins were prominently expressed at both the transcriptional and protein levels in the endoplasmic reticulum protein processing pathway, while malate dehydrogenase played an extremely critical role in cysteine and methionine metabolism pathway. Collectively, these findings indicate that the microtubule cytoskeleton is involved in regulating PCD during pollen cryopreservation, with cysteine and methionine metabolism and endoplasmic reticulum protein processing serving as the core pathways.