免疫组化二抗试剂盒（小鼠/兔超敏聚合物法检测系统）

Pseudomonas aeruginosa pneumonia poses a significant therapeutic challenge. Nanoparticles serve as an effective tool for nucleic acid delivery to efficiently alleviate pneumonia. This study develops a hyaluronic acid (HA)-coated peptide nanoparticle system for targeted delivery of small interfering RNA (siRNA) against Tudor domain-containing protein 9 ( TDRD9 ), identified via RNA sequencing of bronchoalveolar lavage fluid-derived neutrophils from 21 recruited patients (11 males/10 females). Adoptive transfer of TDRD9-silenced polymorphonuclear neutrophils into neutrophil-depleted male mice attenuates lung inflammation and edema. Mechanistically, TDRD9 suppresses neutrophil cuproptosis by upregulating programmed death ligand 1 (PD-L1) through interaction with CD80 to activate p38 mitogen-activated protein kinase (MAPK) signaling. HA-si-TDRD9 nanoparticles enhance neutrophil cuproptosis, reduce pulmonary neutrophil accumulation, and ameliorate lung injury via PD-L1/CD80/MAPK. Importantly, HA-si-TDRD9 nanoparticles reduce bacterial growth, apoptosis, and inflammation in human lung organoids. This work demonstrates that targeting TDRD9 with siRNA nanoparticle platform presents a promising therapeutic strategy for treating bacterial lung injury.

Background Ambra1 has recently been identified as a key regulatory factor in the progression of mantle cell lymphoma (MCL). The objective of this study was to investigate the biological role and molecular mechanism of Ambra1 in MCL. Methods The m6A modification level of Ambra1 was detected by MeRIP-qPCR. Wild-type and mutant Ambra1 plasmids were constructed to verify the direct regulation of Ambra1 by METTL3-mediated m6A modification. The influence of METTL3/m6A/YTHDF2/Ambra1 on the viability, proliferation, migration, apoptosis, and cell cycle of MCL cells was evaluated by standard in vitro assays. RIP and RNA pull-down assays were performed to validate Ambra1 as a downstream target of YTHDF2. Xenograft tumor models were established using BALB/c nude mice to confirm the in vivo phenotype of METTL3 and Ambra1 silencing. Results Ambra1 was downregulated in MCL cells by METTL3-mediated m6A modification. Furthermore, knocking down METTL3 in the MCL cells inhibited their proliferation, migration, and invasion through the upregulation of Ambra1, while METTL3 overexpression had the opposite effect. The m6A reader protein YTHDF2 downregulated Ambra1 expression by binding to Ambra1-m6A. YTHDF2 knockdown inhibited the growth of MCL cells through Ambra1, while YTHDF2 overexpression had the opposite effect. Mechanistically, METTL3 downregulated Ambra1 in the MCL cells in an m6A-YTHDF2-dependent manner to inhibit apoptosis. Finally, METTL3 knockdown inhibited MCL progression in vivo by inducing Ambra1 expression. Conclusion METTL3 promotes MCL progression through YTHDF2-mediated degradation of Ambra1 mRNA, suggesting that the METTL3/YTHDF2/Ambra1 may serve as a potential therapeutic target for MCL.

Background PARP inhibitor (PARPi) maintenance therapy significantly extends progression-free survival of patients with homologous recombination repair deficiency (HRD) or BRCA mutations in ovarian cancer. However, more than 50% of patients lack HRD, highlighting the need to expand PARPi use for homologous recombination -proficient patients. In this study, the efficacy of GX15-070 combined with niraparib in ovarian cancer was evaluated. Methods Based on the core regulators of genome stability and homologous recombination (HR) repair pathway, a compound library was constructed. The effect of candidate drugs on niraparib sensitivity were measured using CCK-8 in ovarian cancer cell lines. Immunofluorescence and non-homologous end joining repair (NHEJ) assay were conducted to examine HR and NHEJ activity. Co-immunoprecipitation was used to investigate the interaction between Mcl1 and Ku70. BH3 domain deletion mutant of Mcl1 was generated to elucidate the structural basis of the interaction between Mcl1 and Ku70. Additionally, cell line-derived xenograft (CDX) and patient-derived xenograft (PDX) mouse models were established to evaluate the efficacy of GX15-070 combined with niraparib in vivo. Results We constructed a compound library based on the core regulators of genomic stability and HR repair. Through high-throughput drug screening, GX15-070, a Mcl1 inhibitor, was identified as a synergist of niraparib, independent of BRCA status. Inhibition of Mcl1 expression significantly impaired HR activity and potentiated niraparib sensitivity. High expression of Mcl1 was associated with a wore prognosis in ovarian cancer patients treating PARPi maintenance therapy. Mechanistically, Mcl1 directly interacts with Ku70 protein via its BH3 domain, serving as a functional switch in selecting between HR and NHEJ. GX15-070 disrupts the interaction by displacing Ku70, promoting a shift in DNA repair pathways from HR to NHEJ. Furthermore, the synergistic efficacy of the combination treatment was further validated in CDX and PDX models. Conclusions The study demonstrated that the combination of GX15-070 with niraparib might be a promising therapeutic strategy for ovarian cancer patients with limited PARPi response.

Breast cancer is associated with a higher incidence of depression and decreased quality of life. Previous studies have indicated that quercetin can mitigate the advancement of breast cancer-related depression (BCRD); however, the specific mechanism by which quercetin affects BCRD is yet to be determined. In this study, we aimed to examine the effect of quercetin on BCRD and explore the underlying mechanisms. We established a mouse model of BCRD and administered quercetin. LC–MS was used to analyze and determine distinct alterations in metabolites in mouse tumor samples. Polymorphonuclear neutrophils (PMNs) were extracted from mouse femurs and treated with PMA and quercetin/Sphingosine 1-phosphate (S1P). Mouse breast cancer cells 4 T1 were treated with lipopolysaccharides (LPS), neutrophil extracellular traps (NETs) and S1P. Neuronal cells were treated with LPS, NETs, S1P, and Corticosterone. Pearson's correlation coefficient was used to evaluate the relationship between differential metabolites and NETs. Quercetin inhibited NET formation in BCRD mice. In vitro, quercetin reversed NET-induced 4 T1 cell proliferation, migration, and ROS production. Quercetin also reversed the effects of NET-induced 4 T1 cells on neuronal cells. LC–MS analysis demonstrated that quercetin ameliorated the metabolic abnormalities in the tumors of BCRD mice. Pearson's correlation analysis showed that S1P, Oleoyl glycine, N-Arachidonoylglycine, 2, 3-butanediol apiosylglucoside, and tetracosatetraenoyl carnitine levels positively correlated with MPO DNA levels. Furthermore, in vitro, S1P enhanced NET-induced 4 T1 cell proliferation, migration, and ROS production, as well as enhanced NET-induced 4 T1 cell damage to neuronal cells. Quercetin alleviated BCRD by inhibiting NETs via inhibition of the S1P/S1PR axis.

Exposure to fine particulate matter (PM 2.5 ) represents a critical environmental health threat, with growing evidence linking it to accelerated chronic kidney disease (CKD) progression. However, the underlying mechanism of this toxicity remains poorly understood. This study investigated whether PM 2.5 exposure induces renal tubular cell senescence and explored the molecular basis of this process. We found that PM 2.5 exposure caused kidney injury in mice and upregulated senescence markers in both mice and human kidney proximal tubule epithelial (HK-2) cells. Mechanistically, PM 2.5 downregulated FOXP1 expression, relieving its transcriptional repression of CDKN1A (encoding P21), leading to P21 upregulation and subsequent cell cycle arrest. Overexpressing FOXP1 or treating with quercetin mitigated PM 2.5 -induced senescence in HK-2 cells. Our findings demonstrate that reduced FOXP1 drives cellular senescence in PM 2.5 -induced renal injury and identify quercetin as a potential therapeutic agent that activates FOXP1 and alleviates PM 2.5 nephrotoxicity.

Introduction: Knee osteoarthritis (KOA) is a degenerative joint disease characterized by the progressive deterioration of cartilage and synovial inflammation. A critical mechanism in the pathogenesis of KOA is impaired efferocytosis in synovial tissue. The present study aimed to identify and validate key efferocytosis-related genes (EFRGs) in KOA synovial tissue by using comprehensive bioinformatics and machine learning approaches.Methods: We integrated three datasets (GSE55235, GSE55457, and GSE12021) from the Gene Expression Omnibus database to screen differentially expressed genes (DEGs) associated with efferocytosis and performed weighted gene co-expression network analysis. Subsequently, we utilized univariate logistic regression analysis, least absolute shrinkage and selection operator regression, support vector machine, and random forest algorithms to further refine these genes. The results were then inputted into multivariate logistic regression analysis to construct a diagnostic nomogram. Public datasets and quantitative real-time PCR experiments were employed for validation. Additionally, immune infiltration analysis was conducted with CIBERSORT using the combined datasets.Results: Analysis of the intersection between DEGs and EFRGs identified 12 KOA-related efferocytosis DEGs. Further refinement through machine learning algorithms and multivariate logistic regression revealed UCP2, CX3CR1, and CEBPB as hub genes. Immune infiltration analysis demonstrated significant correlations between immune cell components and the expression levels of these hub genes. Validation using independent datasets and experimental approaches confirmed the robustness of these findings.Conclusions: This study successfully identified three hub genes (UCP2, CX3CR1, and CEBPB) with significant expression alterations in KOA, demonstrating high diagnostic potential and close associations with impaired efferocytosis. These targets may modulate synovial efferocytosis-related immune processes, offering novel therapeutic avenues for KOA intervention.

Background: While programmed death-ligand 1 (PD-L1)-targeted immunotherapy represents an advancement in non-small cell lung cancer (NSCLC), patient outcomes remain suboptimal. Aberrant activation of the cyclic adenosine monophosphate (cAMP) response element binding protein (CREB)-regulated transcription coactivator (CRTC) is linked to malignant proliferation and functionality in lung cancer cells. This study investigates the involvement of CRTC1 in tumor immunity.Methods: CRTC1 and Notch1 expression were regulated in A549 and NCI-H1299 NSCLC lines through plasmid-mediated overexpression/silencing to assess their effects on cell viability, apoptosis, migration, and invasion. CRTC1/Notch1-dysregulated Lewis lung carcinoma (LLC) cells were co-cultured with T cells to evaluate T cell activation and function. The efficacy of combined CRTC1 knockdown/overexpression and atezolizumab (anti-PD-L1) was tested in an LLC xenograft mouse model.Results: CRTC1 promoted cell viability, migration, and invasion while suppressing apoptosis across NSCLC models. In LLC cells, CRTC1 upregulated tumor cell PD-L1 expression, suppressed T cell-derived IFN-γ and IL-2 production, diminished endogenous CXCL10/11 secretion, and impaired T cell proliferation and cytotoxicity. Mechanistically, CRTC1 interacted with Notch1 to activate the Notch1/Akt pathway, stimulating PD-L1 upregulation, thereby facilitating tumor immunosuppression and growth. Notably, CRTC1 overexpression reversed the protective effects of atezolizumab on tumor growth. Combining CRTC1 knockdown with atezolizumab synergistically enhanced anti-tumor T cell immunity, achieving the most significant tumor regression in xenografts.Conclusion: These findings indicate that CRTC1 in tumor cells suppresses PD-L1-mediated anti-tumor immunity and promotes tumorigenesis via the Notch1/Akt signaling axis. Dual targeting of CRTC1 and PD-L1 demonstrates therapeutic synergy, suggesting CRTC1 pathway inhibition could optimize immunotherapy outcomes in NSCLC patients.

Pancreatic cancer is highly challenging, with most patients developing intrinsic or acquired resistance to first-line chemotherapy drug gemcitabine (GEM). Although Matrix Metalloproteinase 28 (MMP28) is upregulated in pancreatic cancer and predicts a poor prognosis, its role in GEM resistance and molecular mechanism remain unclear. Here, we aimed to investigate the role of MMP28 in GEM resistance and molecular mechanism. First, differentially expressed genes in pancreatic cancer were identified through bioinformatics and validated in clinical samples and cells. MMP28 was significantly overexpressed in pancreatic cancer tissues and Capan-1 and PANC-1 cells, correlating with poor prognosis. Then, MMP28 knockdown was performed in Capan-1 and PANC-1 cells, followed by GEM treatment. Furthermore, in vivo experiments evaluated GEM sensitivity after MMP28 knockdown. The results showed that MMP28 knockdown enhanced GEM sensitivity both in vitro , reducing cell proliferation and survival, and in vivo , where tumor growth was significantly suppressed. Additionally, glycolysis-related changes were assessed. We revealed that glycolysis was implicated as a key pathway in this process, with reduced glucose uptake and lactate production observed after MMP28 knockdown. Protein-protein interaction analysis identified Staphylococcal nuclease domain-containing protein 1 (SND1) as a key interactor, and SND1 expression was upregulated in pancreatic cancer tissues. Moreover, MMP28 interacted with SND1 to regulate SND1′s recruitment of HK2 mRNA to promote glycolysis. However, overexpression of SND1 reversed the effects of MMP28 knockdown, restoring glycolysis and GEM resistance. In conclusion, MMP28 promoted tumor growth and GEM resistance in pancreatic cancer by regulating glycolysis via interaction with SND1.

Neutrophil extracellular traps (NETs) contribute to chronic obstructive pulmonary disease (COPD) pathogenesis by amplifying airway inflammation. Gasdermin D (GSDMD)-mediated pyroptosis is a critical driver of COPD progression. This study provides insights into COPD pathogenesis and provides a theoretical basis for potential therapeutic targets. Mice were exposed to cigarette smoke (CS) for 16 weeks to establish a COPD model. In vitro , alveolar macrophages (AMs) (MH-S) and alveolar epithelial cells (MLE-12) were treated with cigarette smoke extract (CSE). Subsequently, NETs were isolated from phorbol-12-myristate-13-acetate (PMA)-stimulated neutrophils. Lung histopathology, inflammatory markers, and pyroptosis-related proteins were analyzed. Co-immunoprecipitation analysis was used to verify the binding of GSDMD and ubiquitin molecules in cells. Interventions included DNase1 to degrade NET and GSDMD knockdown. In CS-exposed mice, NETs increased the levels of proinflammatory cells and mediators, and lung structure was further disrupted. Pyroptosis of AMs was increased, while phagocytosis of AMs was inhibited. However, treatment with DNAse1 partially reversed the results caused by CS exposure and NET induction. Consistently, NETs aggravated inflammatory response and pyroptosis in the CSE-induced MH-S cell model. Furthermore, NETs significantly caused an increase in ROS, which promoted the activation of GSDMD deubiquitination and subsequent pyroptosis pathway in AMs. DNase1 treatment or GSDMD silencing attenuated pyroptosis, reduced inflammatory mediators, and improved lung function. NETs aggravated CS-induced lung inflammation and injury by activating GSDMD to promote pyroptosis in AMs. Targeting GSDMD or NETs represents a novel therapeutic strategy for COPD. © 2026 The Pathological Society of Great Britain and Ireland.

Aims Myocardial hypertrophy is a key pathological basis for heart failure, closely related to disturbances in cardiac lipid metabolism and impaired mitochondrial function. Long-chain acyl-CoA synthetase 6 (ACSL6) is a pivotal enzyme in fatty acid metabolism, but its role in myocardial hypertrophy remains unclear. This study aims to investigate the role of ACSL6 in myocardial hypertrophy and its potential mechanisms. Materials and methods Mouse models of myocardial hypertrophy induced by isoproterenol (ISO) and neonatal mouse cardiomyocyte (NMCM) hypertrophy models intervened by angiotensin II (Ang II) were established. Using lentivirus-mediated ACSL6 overexpression, co-immunoprecipitation, mass spectrometry, lipidomics, transmission electron microscopy, and molecular biology techniques, the functions and mechanisms of ACSL6 were explored. Key findings ACSL6 was downregulated in ISO-induced myocardial hypertrophic mouse tissues and Ang II-treated NMCMs, with expression decreasing as Ang II intervention duration increased. ACSL6 overexpression significantly alleviated myocardial hypertrophy, improved cardiac function, and mitigated cell damage and hypertrophic marker upregulation. Additionally, ACSL6 overexpression inhibited myocardial lipid synthesis and accumulation, ameliorated lipid metabolic disorder, and enhanced mitochondrial function in ISO-induced mice. Mechanistically, KRT17 bound to ACSL6, competing with E3 ubiquitin ligase MIB1, protecting ACSL6 from ubiquitination and degradation. KRT17 knockdown reversed the protective effects of ACSL6 overexpression, exacerbating lipid accumulation and mitochondrial dysfunction. Significance ACSL6 alleviated myocardial hypertrophy by ameliorating cardiac lipid synthesis and mitochondrial function. KRT17 stabilized ACSL6 expression by inhibiting its ubiquitination and degradation, mediating ACSL6's protective effects. Targeting the KRT17-ACSL6 axis emerged as a promising strategy for treating myocardial hypertrophy.