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.

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.

Starch degradation is a fundamental process underlying the development of key postharvest quality attributes in ripening mango fruit; however, its regulatory mechanisms remain incompletely understood. In this study, ethephon (ETH) and 1-methylcyclopropene (1-MCP) were used as ripening modulators to investigate the patterns of starch degradation and its transcriptional regulation in postharvest mango fruit. The results demonstrated that ETH fumigation accelerated the ripening process, while 1-MCP treatment effectively inhibited this process. Transcriptome and RT-qPCR analyses revealed and validated that six starch-degrading genes (MiAMY1.1, MiAMY3, MiBAM3, MiBAM9, MiGWD and MiPWD) were regulated in response to ETH. Furthermore, the transcription factor MiRAP2.11 was identified as a nucleus-localized transcriptional activator of MiBAM3 and MiPWD. Transient transformation assays demonstrated that MiRAP2.11 positively regulated starch degradation, thereby promoting development of ripening and quality attributes. The results indicate that MiRAP2.11 functions as a crucial regulator mediating ethylene-induced starch degradation during mango ripening.

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.

This study is aimed to elucidate the role of ubiquitin-specific proteases (USP) 15 in cerebral ischemia-reperfusion injury (CIRI) under the influence of sevoflurane (Sev).

Gastric cancer (GC) is one of the most prevalent gastrointestinal malignancies and continues to pose a significant global health burden. Repurposing clinically approved drugs is a cost-effective strategy for identifying new anticancer agents. In this study, the anticancer effects of linagliptin (LG), a dipeptidyl peptidase-4 (DPP-4) inhibitor, were evaluated in the human gastric cancer cell line HGC-27. LG significantly reduced cell viability in a dose-dependent manner, with a half-maximal inhibitory concentration (IC50) value of 50.29 μM, and increased apoptotic cell death as confirmed by flow cytometry. Gene expression analysis revealed significant downregulation of Smoothened (SMO) and split hand and foot malformation 1 (SHFM1) and upregulation of Sonic hedgehog (SHH), indicating modulation of Hedgehog signaling. In addition, LG treatment resulted in reduced global DNA methylation levels. Molecular docking analyses demonstrated favorable binding affinities between LG and key Hedgehog pathway proteins, supporting a potential mechanistic basis for the observed biological effects. Collectively, these findings suggest that LG exhibits antiproliferative and pro-apoptotic activity in gastric cancer cells and may represent a promising candidate for drug repurposing in gastric cancer therapy.

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.

To combat the growing challenge of bacterial infections, sonodynamic antibacterial technology has gained increasing attention. However, current approaches still face limitations, including suboptimal efficacy and a narrow antibacterial spectrum (only targeting a single bacterial species). To overcome these drawbacks, this study designed a composite sonosensitizer, HMME@AMP, by conjugating hematoporphyrin monomethyl ether (HMME) with the antimicrobial peptide LL-37. This composite achieves efficient inhibitory effects against representative Gram-negative and Gram-positive bacteria through sonodynamic therapy (SDT). Using optimized ultrasound parameters (0.5 W/cm2, 1 MHz, 60% duty cycle), we evaluated the antibacterial activity and mechanisms of HMME@AMP through agar culture, flow cytometry, bacterial weight measurement, scanning electron microscopy (SEM), whole-genome sequencing, and quantitative real-time PCR (qPCR). Results demonstrated that HMME@AMP exhibited strong, concentration-dependent antibacterial effects against Pseudomonas aeruginosa (P. aeruginosa) and Staphylococcus aureus (S. aureus) under ultrasound irradiation. At a concentration of 120 μg/mL, bacterial weight measurement results showed that the bacterial weight inhibition rates reach 56.4% (for P. aeruginosa) and 87.3% (for S. aureus), while flow cytometry indicated survival rates below 3 and 7%, respectively, confirming its excellent inhibitory ability against P. aeruginosa and S. aureus. SEM imaging revealed ultrasound-induced nanoscale pores and membrane collapse, indicating combined physical damage and ROS-mediated oxidative stress. Genome sequencing and qPCR further showed that HMME@AMP downregulated critical genes in P. aeruginosa (e.g., PA0876, PA4896, involved in phenazine synthesis and iron homeostasis) and S. aureus (e.g., SAOUHSC-02494, linked to ribosome function), disrupting bacterial metabolism and proliferation. This study demonstrates that HMME@AMP-mediated SDT achieves potent antibacterial effects through dual mechanismsphysical disruption and genetic regulationoffering a promising, antibiotic-free strategy for treating deep-tissue infections.

Acinetobacter baumannii, a major pathogen that causes nosocomial infections, exhibits drug resistance. Efflux pumps are critical in developing multidrug resistance. Berberine hydrochloride exhibited inhibitory effects against bacteria and toxins, demonstrating antibacterial efficacy. However, there is limited research on berberine hydrochloride combined with multiple antibiotics through regulation of efflux pumps in multidrug-resistant A. baumannii. We evaluated the antibacterial effect of berberine hydrochloride in combination with multiple antibiotics, explored the resistance mechanisms regulated by efflux pumps, and analyzed the differential expression of active efflux pump genes (adeR, adeS, adeJ, adeM, adeG, and adeB) before and after berberine hydrochloride exposure, to provide new therapeutic strategies for managing multidrug-resistant A. baumannii infections. One hundred non-repetitive A. baumannii strains were isolated from patients with respiratory infections. The microdilution method was used for the determination of minimal inhibitory concentration (MIC) and the checkerboard method for the determination of the fractional inhibitory concentration (FIC) index. Reverse transcription polymerase chain reaction quantified efflux pump gene expression in sensitive and resistant strains before and after berberine hydrochloride treatment. Strains showing significant differences were selected for transcriptomic sequencing. Berberine hydrochloride showed synergistic effects with nine antibiotics, the strongest being amikacin, meropenem, and cefepime. Resistance reversal occurred especially with levofloxacin. adeJ and adeB expression differed significantly between sensitive and resistant strains. Berberine hydrochloride induced notable changes in adeS, adeG, adeB, and adeR. Transcriptome sequencing revealed the significant expression of adeA, adeN, and adeT2. Berberine hydrochloride enhances antibiotic sensitivity and reverses resistance by regulating efflux pump gene expression, providing a theoretical foundation for improved treatment.IMPORTANCEThis study investigates multidrug-resistant Acinetobacter baumannii (MDRAB) strains isolated from clinical infections using the microplate dilution method to evaluate the antibacterial efficacy of berberine in combination with various antibiotics. The research simultaneously analyzes the differential expression levels of active efflux pump genes before and after berberine treatment. The findings aim to identify novel therapeutic strategies for MDRAB, explore the drug resistance mechanisms mediated by efflux pumps in AB, and elucidate berberine's role in reversing AB resistance through efflux pump regulation. These insights provide theoretical foundations for improving cure rates in MDRAB-infected patients and advancing drug target design.

Dengue virus (DENV) remains a major global health concern without effective treatments. Previously, we identified sulfonyl anthranilic acid (SAA) derivatives (compounds 1 and 2) as potent pan-DENV inhibitors, likely targeting a primate-specific factor. Here, mass spectrometry-based target deconvolution revealed that SAA compounds downregulate ribosomal protein expression, some of which are essential for DENV replication, as confirmed by siRNA-knockdown studies. This novel mechanism aligns with the broad-spectrum antiviral activity of compounds 1 and 2. Moreover, compound 1 was also effective against the Zika virus in a human brain organoid model. The subsequent medicinal chemistry optimization process resulted in the identification of compound 7, which demonstrated an EC50 value of 50 nM against DENV-2, promising broad-spectrum potential and favorable in vitro ADME properties. Further studies indicated that these compounds modulate the 5'-terminal oligopyrimidine (5'-TOP) motif in ribosomal mRNAs. These findings open a new avenue for antiviral development by targeting a previously unexplored host pathway.

As an inhibitor of DNA methyltransferases (DNMTs) and an anti-cancer drug, 5-aza-cytidine (5-azaC)'s many effects on gene expression remain unclear. Here, we show that 5-azaC treatment of cultured GH3 pituitary tumor cells increases relative usage of genomic terminal exons (GTEs) across the transcriptome. This effect is largely achieved by shifting mRNA polyadenylation from proximal poly(A) sites to GTEs, which harbor a more optimal consensus motif of poly(A) signals. Consistent with this shift, 5-azaC upregulates the mRNA anti-termination factors Scaf4 and Scaf8 while downregulating the early termination enhancer E2f2. In MOLM-13 leukemia cells, 5-azaC similarly promotes the production of full-length transcripts and regulates alternative polyadenylation factors, some of which are in the same direction as observed in GH3 cells. Moreover, PCF11, a factor known to promote proximal poly(A) site usage, is upregulated in both cell lines, suggesting a homeostatic response by these cells to counteract transcript lengthening during 5-azaC treatment. Together, these findings uncover a previously unknown effect of 5-azaC on gene expression: directional promotion of terminal polyadenylation site usage, driving a transcriptome-wide switch from shortened to full-length mRNAs in tumor or cancer cells and consequently altering the alternative usage of multiple 3' exons.

Current toxicology and cancer biology investigations have focused on developing alternative models that better recapitulate the in vivo architecture of tissues and organs. The present study evaluated the anticancer effects of the flavone cirsimarin, which presented successful antitumor activity on breast tumor cells. We assessed the impact of flavone on cell viability, proliferation, and migration, as well as on DNA integrity and modulation of related cellular pathways. In the 2D model, cirsimarin reduced cell viability at concentrations ≥ 80 μM after 24 h of treatment (resazurin assay), selectively in A549 cells compared to MRC-5 non-tumor cells. Apoptosis was induced at concentrations ≥ 40 μM, and clonogenicity was reduced by approximately 50% only at 160 μM. In the wound healing assay, cirsimarin (1-80 μM) completely inhibited cell migration and induced DNA damage (comet assay). These apoptotic and anti-migratory effects were associated with the downregulation of key genes involved in cell proliferation, death, and extracellular matrix remodeling, including TNF-α (0.32-fold), TP53 (0.17-fold), MMP-2 (0.18-fold), MMP-9 (0.43-fold), and MMP-11 (0.04-fold), as revealed by RT-qPCR analysis. In the 3D model, after 216 h of treatment, cirsimarin reduced cell viability (≥ 40 μM) and spheroid area (≥ 80 μM) while antimigratory effects were observed only in the highest concentration evaluated (160 μM). These findings could indicate a potential reduction in lung tumor growth and metastasis, warranting further investigation, particularly of the antimetastatic effect of this flavone.

Rosmarinus officinalis (Ro), popularly known as rosemary, is an aromatic perennial shrub originated from Mediterranean region and belongs to family Lamiaceae. Although the health promoting effects of Ro in dermatology, especially in wound healing and hair loss were documented to a certain extent, specific studies in this field are lacking. In this study it is investigated the molecular bases of these cutaneous effects, by determining gene expression levels of VEGF and IL-1α, in a human keratinocyte cell line (HaCaT) treated with Ro extract.

Alfalfa (Medicago sativa L.) is a globally important forage legume and the most widely cultivated sown pasture species in China. Drought, as one of the most common abiotic stresses, limits alfalfa growth and development. Nitric oxide (NO), a key signaling molecule, plays an essential role in plant growth, development, and responses to various abiotic stresses. In this study, exogenous NO was applied to alfalfa seedlings under drought stress, followed by physiological and transcriptomic analyses. The results showed that sodium nitroprusside (SNP)-treated alfalfa seedlings grew better than untreated controls (CK), with improved leaf tissue structure. Meanwhile, SNP treatment increased proline content, reduced malondialdehyde accumulation, and enhanced hydroxyl radical scavenging capacity. Under drought stress, lignin content increased in alfalfa seedlings. Following exogenous NO application, lignin content in alfalfa seedlings further increased. RNA-Seq analysis identified 20,183 differentially expressed genes (DEGs) in alfalfa seedlings treated with PEG, SNP, or PEG + SNP. KEGG enrichment analysis indicated that the DEGs under drought stress were involved in the phenylpropanoid biosynthesis pathway, which regulates lignin biosynthesis and enhances drought tolerance. GO enrichment analysis revealed that these DEGs participated in the response to water deprivation, thereby modulating drought stress tolerance and improving drought resistance. Furthermore, we assessed the transcript-level changes in genes induced by phenylpropanoid biosynthesis in alfalfa. Among them, 124 DEGs were identified as participating in phenylpropanoid biosynthesis, including 10 up-regulated DEGs (three of which encode key enzymes associated with lignin synthesis), while the remaining DEGs were down-regulated. These findings provide new insights into the transcriptomic mechanisms of SNP-mediated drought adaptation in alfalfa and reveal key pathways contributing to drought tolerance in this species.

Large-scale randomized trials have found that multivitamin-multimineral (MVM) supplements and cocoa flavanols may benefit several age-related chronic conditions among older adults, but it remains unclear whether these two supplements directly slow the biological aging process. This prespecified ancillary study evaluated the 2-year effect of a daily MVM (Centrum Silver) and cocoa extract (500 mg cocoa flavanols per day, including 80 mg (-)-epicatechin) on five DNA methylation measures of biological aging (PCHannum, PCHorvath, PCPhenoAge, PCGrimAge and DunedinPACE) among 958 participants (482 women and 476 men) in the COcoa Supplement and Multivitamin Outcomes Study (COSMOS). Compared with placebo, daily MVM supplementation modestly reduced the rate of increase of second-generation epigenetic clocks, with a between-group difference in yearly change of -0.113 years (95% confidence interval (CI) -0.205 to -0.020; P = 0.017) for PCGrimAge and -0.214 years (-0.410 to -0.019; P = 0.032) for PCPhenoAge. MVM had a stronger effect on PCGrimAge among those with accelerated biological aging at baseline (-0.236 [-0.380 to -0.091]) compared with those with normal or decelerated biological aging (-0.013 [-0.130 to 0.104]; P = 0.018 for interaction). Cocoa extract did not have an effect on the five epigenetic clocks tested. Although the statistically significant but small effects of daily MVM supplementation on slowing biological aging are encouraging, additional studies are needed to determine the clinical relevance of daily MVM supplementation on epigenetic clocks and whether such effects can help explain the beneficial effects of MVM supplementation on aging-related chronic conditions.