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.

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.

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.

Multiple myeloma (MM) is an incurable plasma cell neoplasm that is highly reliant on endoplasmic reticulum-associated degradation (ERAD) to maintain protein homeostasis. Disrupting ERAD has been proposed as a therapeutic strategy to overcome proteasome inhibitor resistance; however, the identification of novel inhibitors has been limited. To address this, we conducted a cell-based high-throughput screen using the FDA repurposing library and identified omaveloxolone (RTA408) as a potent ERAD inhibitor that selectively impairs the degradation of ER luminal and membrane substrates, without affecting the degradation of key cytosolic proteins that are implicated in disease relapse. Surprisingly, although ER stress response pathways are activated after ERAD inhibition in MM, we find that apoptosis is mediated by altered lipid raft organization, leading to aberrant activation of the death-inducing signaling complex (DISC) and caspase 8 in the extrinsic apoptotic pathway. Notably, ERAD inhibition by RTA408 is cytotoxic to primary malignant plasma cells, including those resistant to proteasome inhibitors, and demonstrates in vivo anti-myeloma activity. Our findings establish a novel ERAD inhibitor, which is a valuable tool to dissect ERAD biology, and provide pre-clinical evidence for RTA408 as a therapeutic agent in MM.

The treatment of colorectal cancer (CRC) remains challenging due to chemotherapy resistance and genetic heterogeneity. Indole-3-lactic acid (ILA), a tryptophan metabolite derived from gut microbiota, exhibits promising anti-inflammatory and anticancer properties; however, its specific molecular targets and regulatory mechanisms in CRC remain poorly understood. In this study, we combined network pharmacology and machine learning with molecular docking to identify candidate targets and pathways for ILA in CRC. We identified 39 ILA-CRC common targets, ultimately identifying four hub genes through the intersection of machine learning models. Validation in independent GEO datasets confirmed significant differential expression of these genes in CRC tissues. Functional enrichment analyses linked these genes to the PPAR, PI3K-AKT, and IL-17 signaling pathways, and gene set enrichment analysis further implicated ascorbate and aldarate metabolism, DNA replication, and fatty acid metabolism. Immune infiltration analysis indicated associations between hub gene expression and immune cell populations, including mast cells, neutrophils, and macrophages, suggesting potential involvement in the tumor immune microenvironment. Molecular docking supported favorable binding of ILA to all four hub proteins, and 100-ns molecular dynamics simulations specifically validated the dynamic stability of the ILA-HMOX1 complex. In conclusion, these results highlight EPHA2, HMOX1, MMP3, and PARP1 as candidate targets and suggest that ILA may influence CRC-related signaling, metabolic programs, and immune contexture, providing a theoretical foundation for developing gut microbiota-derived metabolites as novel anticancer strategies.

Glucocorticoids (GCs) are steroid hormones that bind to the glucocorticoid receptor (GR) as ligands to initiate systemic anti-inflammatory effects. GCs are commonly administered alongside chemotherapy to reduce treatment-related side effects in breast cancer patients. However, GC administration has been shown to promote metastasis in breast cancer. In this study, we used quantitative mass-spectrometry-based approaches to analyze proteome and phosphoproteome of three breast cancer cell lines following treatment of a clinically approved synthetic GC, dexamethasone (Dex). By comparing MCF7, MDA-MB-231, and MDA-MB-436 cells, we suggest that the level of GR significantly affects Dex-mediated responses. Additionally, we identify noncanonical transcription factors (TFs) and kinases that are regulated by GR in different cell lines. Together, our data present Dex-induced protein modulations and modifications involving several TFs and kinases that regulate cytoskeletal remodeling and migration in breast cancer cell lines. These findings highlight the need for careful consideration of GC use in breast cancer therapy and identify potential molecular targets for mitigating adverse effects.

Cell surface receptors play vital roles in cancer growth and metastasis. Integrin αvβ3 is overexpressed in various cancer cells and interacts with different growth factors to stimulate cancer progression. Thyroid hormone binds to αvβ3 to activate signal transduction and cell proliferation. However, thyroxine (T4) deaminated analogue, tetraiodothyronine (tetrac), competes for the binding on integrin and inhibits cancer cell growth and metastasis. The current study investigated the pathogenic role of integrin αvβ3 and the potential of a novel therapeutic strategy targeted to integrin αvβ3. Pathogenetic studies of clinical samples revealed integrin αvβ3 cross-talked with EGFR and downstream signal transduction networks affected by thyroid hormone and EGF related to the progression of cholangiocarcinoma malignancy. Thyroxine and EGF stimulated PD-Ligand 1 (PD-L1) expression and cancer growth in cholangiocarcinoma. The thyroxine-induced PD-L1 accumulated in the nuclei and colocalized with p300. Alternatively, EGF increased cytosolic PD-L1 and nuclear accumulation of β-catenin. Targeting integrin αvβ3 with lipo-tetrac and its Dox-derivative induced anti-proliferation in vitro and in the xenografted animal model. Our research provides a fundamental understanding of the therapeutic role of integrin αvβ3 and the potential therapeutic approach in cholangiocarcinoma treatment.

Triple-negative breast cancer (TNBC) is an aggressive subtype lacking targetable hormone receptors, making conventional chemotherapy the primary treatment option, despite its associated toxicity and potential for drug resistance. Melatonin, a natural hormone with anticancer and immunomodulatory properties, has shown promise in multiple cancers; however, its role in TNBC remains unclear.

Prostate cancer (PCa) is the most prevalent malignancy in men and often correlates with distant metastasis in its advanced stages. The study aimed to investigate the effects of Ugonin J, a natural compound isolated from Helminthostachys zeylanica, on PCa metastasis.

B-cell lymphoma 6 (BCL6) is a transcriptional repressor whose overexpression is closely linked to the progression of diffuse large B-cell lymphoma (DLBCL), making it a promising therapeutic target. This study aims to identify a novel small molecule, synthesized via proteolysis-targeting chimeras (PROTACs), capable of degrading BCL6, thereby inhibiting DLBCL growth and providing a foundation for future preclinical studies.

Tamoxifen is a key drug that provides endocrine therapy for estrogen receptor (ER) α-positive breast cancer; however, resistance remains a significant clinical challenge. This study aims to investigate the molecular mechanisms of tamoxifen resistance in ERα-positive breast cancer, with particular focus on the role of SET Domain Containing 1A (SETD1A)-driven forkhead box A2 (FOXA2) as a key regulator of this resistance.

Clear cell renal cell carcinoma (ccRCC) is predominantly treated with anti-angiogenic therapies (AATs), such as sunitinib and axitinib. While these therapies initially improve outcomes, resistance frequently emerges, limiting long-term efficacy. Understanding the molecular mechanisms underlying AAT resistance is essential to optimize treatment strategies.

PD-L1 is one of the most critical immune checkpoint proteins, inhibiting T-cell immune responses by binding to PD-1. This study aims to validate that allicin can regulate PD-L1 expression through the IL-6/JAK2/STAT3 pathway, thereby inhibiting immune evasion in osteosarcoma.

Intracerebral hemorrhage (ICH) has high mortality and disability with limited efficient therapies. The inhibition of ferroptosis is implicated in the prognosis of ICH. Epigallocatechin gallate (EGCG) has iron chelating, anti-inflammatory and free radical scavenging properties, which connected with Nrf2-Keap1 pathway in previous researches. Our investigation aimed to identify the role of EGCG on ferroptosis in ICH rats and the underlying mechanisms through Nrf2-Keap1 pathway. In this study we observed that EGCG treatment effectively reduced hematoma volume, cellular destruction of brain tissue and ferroptosis in neuronal cells, and improved neurobehavioral outcomes in rat model of ICH. Our further studies showed that EGCG promoted the expression of nuclear factor erythroid 2-related factor 2 (Nrf2), stimulated Nrf2-Keap1 complex releasing Nrf2 and promoted the translocation of Nrf2 from the cytoplasm to the nucleus, and regulated the downstream ferroptosis-regulated proteins to inhibit ferroptosis, while Nrf2 inhibitor reversed the anti-ferroptosis effect, suggesting that EGCG inhibit ferroptosis by upregulating Nrf2. Moreover, ultilizing the in vitro ICH model, we revealed that when Keap1 is silenced, the regulation of ferroptosis-related mRNA by EGCG through Nrf2 was enhanced, which is direct opposite of the Nrf2 silencing result, indicating that Nrf2-Keap1 pathway plays an important role in EGCG treatment after ICH. Our study confirmed for the first time that EGCG could reduce the iron deposition by regulating downstream ferroptosis-related proteins via Nrf2-Keap1 pathway and inhibit ferroptosis after ICH, indicating that EGCG could be a potential drug for the treatment of ICH.

Cardiac arrest (CA) remains a major public health challenge with high incidence and mortality. Post-cardiac arrest brain injury (PCABI) is the primary determinant of poor neurological outcomes and survival. Although curcumin (Cur) exhibits neuroprotective effects in multiple cerebral injury models, its precise pharmacological mechanisms in PCABI remain incompletely understood.

Hepatocellular carcinoma (HCC) is a primary malignant tumour that impacts patients' quality of life. Currently, clinical experience from The First Affiliated Hospital of Guangzhou University of Chinese Medicine suggests that Jianpi Jiedu Xiaozheng Fang (JPJDXZF) demonstrates promising efficacy in the treatment of HCC. We aimed to explore the mechanisms of JPJDXZF in HCC based on network pharmacology. The components and their relevant targets of JPJDXZF were identified using databases such as SymMap, TCMID, TCMSP, and TCM-ID. Following ADME screening, 1443 active components of JPJDXZF were identified, and 435 corresponding drug targets were predicted using the SwissTargetPrediction database. Subsequently, prognosis-related differentially expressed genes (DEGs) associated with HCC were analyzed using TCGA and GTEx datasets, and a gene expression matrix was derived. Key genes involved in HCC regulation were identified, and functional analyses were performed. Furthermore, we explored the regulatory effects of JPJDXZF at the cellular, organoid, and animal levels. We identified 18 intersecting genes between HCC prognosis-related genes and JPJDXZF-target genes. Venn diagram analysis successfully identified BIRC5 and CYP2E1 as two potential targets for JPJDXZF in treating HCC. Pathway enrichment analysis indicated that the core targets of JPJDXZF were enriched in multiple signalling pathways, including the Hippo pathway, in which BIRC5 is involved as a downstream regulatory gene. In in vitro experiments, JPJDXZF-containing serum significantly reduced the viability and migration of HepG2 and MHCC97-H cells, leading to a decrease in organoid diameter and ATP activity in HCC organoids. In in vivo experiments, tumours in nude mice treated with JPJDXZF exhibited reduced volume and weight, along with decreased expression of BIRC5 and Hippo pathway effectors YAP and TAZ. At the mechanistic level, JPJDXZF treatment was associated with altered Hippo pathway-related signalling, accompanied by reduced YAP/TAZ activity and changes in BIRC5 expression, together with effects on HCC cell proliferation and apoptosis. In addition, siMST1/2 interference and EMT inhibitor-1 treatment partially attenuated the effects of JPJDXZF on cell viability, migration, and apoptosis. JPJDXZF regulates BIRC5 expression in association with Hippo pathway activity in HCC. In vitro, in vivo, and molecular mechanism analyses support JPJDXZF as a potential therapeutic strategy for HCC by modulating key proteins in the Hippo pathway, thus affecting HCC cell proliferation, apoptosis, and migration.

Histone deacetylase 6 (HDAC6) has emerged as a promising therapeutic target in cancer due to its immunomodulatory effects. While its prognostic significance remains debated, we demonstrate that HDAC6 loss significantly impairs myeloid leukemia progression in vivo, despite having no functional impact on leukemia cell proliferation in vitro. Global proteome and secretome profiling of HDAC6-knockout (KO) cells revealed upregulation of several immune-related modulators, including RNase T2, a tumor suppressor known to modulate the tumor microenvironment. Notably, RNase T2 upregulation upon HDAC6 loss was observed in myeloid leukemia cells but not in lymphoblastic leukemia cells. Moreover, pharmacological inhibition of HDAC6 recapitulated this phenotype, leading to RNase T2 upregulation in myeloid leukemia cells. ATAC-seq revealed increased chromatin accessibility of RNase T2 following HDAC6 loss, highlighting a functionally epigenetic regulatory contribution. Further functional assays conducted in an immunocompetent setting, both ex vivo and in vivo, demonstrated that HDAC6 inhibition sensitized murine myeloid leukemia cells to broad CD8+ T cell activation as evidenced by increased TNFα and CD107a expression. Consistently, in a syngeneic murine model, HDAC6 inhibition restricted the growth of myeloid leukemia cells. Moreover, an extended drug screening analysis identified Cytarabine and Clofarabine as significantly synergizing with HDAC6 inhibitor (Ricolinostat) in myeloid leukemia cell lines and in patient-derived xenograft (PDX) cells, while showing limited synergy in lymphoid leukemia cell lines, PDX, or healthy control cells. These findings suggest that HDAC6 represents a promising therapeutic target in myeloid lineage-derived leukemia cells by simultaneously enhancing immune activation and increasing chemosensitivity.

Silicosis remains a critical occupational health concern worldwide, lacking effective treatments due to unclear mechanisms. In this study, we investigated the citrullinated proteomic profile and its effects in mice exposed to silica. Our findings demonstrated elevated levels of citrullinated peptides and citrullinated vimentin (Cit-Vim) in silicotic mice and silica-treated macrophages, regulated by peptidylarginine deiminase (PADI2). Unlike vimentin, Cit-Vim amplified the production of tumor necrosis factor-α (TNF-α), Interleukin-6 (IL-6), and IL-1β in silica-treated macrophages through interaction with Toll-like receptor 4 (TLR4) signaling. RNA sequencing revealed that early growth response protein 1 (EGR1) is a target of PADI2, with Cit-Vim inducing lung inflammation via EGR1 signaling. Pharmacological inhibition or genetic knockout of Padi2 attenuated silica-induced lung inflammation and fibrosis. These findings suggest that targeting PADI2 may represent a novel therapeutic strategy of silicosis.

The aim of the present research is to deeply investigate the cytotoxic and immunomodulatory activities of the mucin extracted from Ereminia desertorum snails´ mucus against two tumor cell lines; human hepatocellular carcinoma (HepG-2) and human colon adenocarcinoma (CACO-2) cells. Both cell lines were treated with Ereminia desertorum snails´ mucin and the anti-cancer potential of the mucin was evaluated by the crystal violet assay test and gene expression analysis using reverse transcription- polymerase chain reaction (RT-PCR).