Potassium (K) supplementation enhances apple (Malus ×domestica) resistance to Cytospora mali, thereby reducing apple Cytospora canker occurrence in the field. Nevertheless, the molecular mechanisms underlying this K-enhanced resistance remain largely unelucidated. Here, we found that brassinosteroid (BR) biosynthesis was active in apple tissues under sufficient K (SK) conditions, resulting in an elevation of brassinolide (BL), the bioactive BR. Exogenous BL application under low K (LK) conditions restored apple resistance in both detached branches and field-grown trees, while the BR inhibitor brassinazole (BRZ) attenuated SK-dependent resistance. Notably, BR biosynthetic gene MdBR6ox2 was upregulated under SK conditions. Silencing of MdBR6ox2 reduced BL content and consequently compromised resistance to C. mali in SK apple calli. Furthermore, we identified that MdMYB306 is responsible for transactivating MdBR6ox2 expression via binding to the MBS cis-element in its promoter. Gene function analysis confirmed the positive role of MdMYB306 in regulating BR biosynthesis. Importantly, K supplementation facilitated the recruitment of MdKIM1, which interacts with MdMYB306 and induces conformational changes in MdMYB306, and enhances its transactivation capacity toward MdBR6ox2. Genetic evidence confirmed the importance of the MdKIM1-MdMYB306 module in initiating BR biosynthesis and increasing resistance in apple under SK status. Collectively, our findings uncovered a MdKIM1-MdMYB306-MdBR6ox2 module that promotes BR accumulation under SK conditions, providing a promising strategy for managing Cytospora canker via combined application K and BR.

BACKGROUND The global prevalence of type 2 diabetes mellitus (T2DM) is increasing. Although globular adiponectin (gAd) shows potential in improving islet function, its clinical application is limited by rapid clearance. Given the promising prospects of adipose-derived mesenchymal stem cell exosomes (Exos) in targeted therapy, whether this nanocarrier can enhance gAd's efficacy in improving islet function warrants significant research attention. AIM To develop a new synergistic therapeutic strategy based on adipose-derived mesenchymal stem cells Exos loaded with gAd (gAd-Exo). METHODS A T2DM rat model was established using a high-fat diet and streptozotocin. Rats were randomized into control, T2DM, T2DM + gAd, T2DM + Exo, and T2DM + gAd-Exo groups, receiving respective treatments via tail vein injection for four weeks. Pancreatic tissues were subjected to histological, immunohistochemical, and biochemical analyses. Meanwhile in vitro experiments assessed the protective effects of gAd-Exo on palmitic acid-injured INS-1 cells. RESULTS gAd-Exo treatment significantly ameliorated hyperglycemia, improved pancreatic islet morphology, and reduced beta-cell apoptosis compared to other groups. It enhanced insulin sensitivity and down-regulated glucagon expression. Mechanistically, gAd-Exo activated the AMP-activated protein kinase/acetyl-CoA carboxylase signaling pathway. In vitro, gAd-Exo superiorly mitigated palmitic acid-induced oxidative stress and apoptosis in INS-1 cells. CONCLUSION This study shows that the combination of gAd and Exo produced a significant synergistic effect. gAd-Exo can relieve type 2 diabetes by reducing blood glucose, improving hyperinsulinaemia and islet function, and at the same time reducing islet beta cells apoptosis. It may be achieved by activating the AMP-activated protein kinase/acetyl-CoA carboxylase pathway. The discovery provides a new strategy with synergistic regenerative potential for diabetes treatment.

Background: Chemotherapeutic resistance remains a major cause of treatment failure in bladder cancer. While the epithelial-mesenchymal transition (EMT) is a known driver of metastasis and drug resistance, its regulatory mechanisms require further elucidation. This study investigates the role of murine double minute 4 (MDM4), a key p53 suppressor, in mediating cisplatin resistance through the EMT program. Methods: A cisplatin-resistant T24 cell line (T24-CR) was established, with MDM4 overexpression confirmed through reverse-transcription Quantitative Real-time PCR (RT-qPCR). Functional assays including colony formation, transwell migration, apoptosis analysis, and western blot were performed following MDM4 knockdown. In vivo validation was conducted using a subcutaneous xenograft model in male BALB/c nude mice divided into wild-type T24 cell line (T24-WT), T24-CR, and MDM4-deficient cisplatin-resistant T24 cells (T24-CR-shMDM4) groups. Results: MDM4 expression was substantially elevated in chemoresistant cells T24-CR at both transcriptional and translational levels. MDM4 depletion effectively restored cisplatin sensitivity, as evidenced by significantly reduced cell viability and colony formation capacity. The knockdown also dramatically impaired metastatic potential, with migration and invasion rates decreasing to levels comparable with parental sensitive cells. At the molecular level, MDM4 inhibition activated the p53 pathway and reversed EMT progression, characterized by E-cadherin restoration and Vimentin downregulation. In vivo studies corroborated these observations, with MDM4-targeted tumors exhibiting slower growth kinetics, prolonged survival outcomes, and histopathological features consistent with mesenchymal-to-epithelial reversion. Conclusions: Our findings identify the MDM4/p53/EMT axis as a pivotal mechanism driving cisplatin resistance in bladder cancer. The study establishes MDM4 as a promising therapeutic target for overcoming chemoresistance, with experimental evidence showing that its inhibition restores p53 activity and reverses the EMT phenotype both in vitro and in vivo.

Purpose Sepsis-induced immunosuppression is a core factor contributing to opportunistic infections and high mortality in patients, with macrophage phagocytic dysfunction being a critical component. Currently, effective therapeutic approaches to reverse macrophage phagocytic capacity remain limited. Patients and methods The study employed an endotoxin-tolerant (ET) macrophage model and a mouse sepsis model induced by cecal ligation and perforation (CLP). The effects of clioquinol (ClioQ) on macrophage phagocytic function, inflammatory cytokine expression, and in vitro/in vivo clearance of fungi (Candida albicans, C. albicans) and bacteria (Escherichia coli, E. coli) were evaluated via flow cytometry-assessed phagocytosis, organ pathogen burden detection, and histopathological analysis. Results In the ET model, ClioQ treatment not only restored TNF-α secretion in response to lipopolysaccharide stimulation but also significantly rescued impaired phagocytosis and intracellular bactericidal functions. In the CLP-induced macrophage phagocytosis impairment model, ClioQ effectively improved macrophage phagocytosis of C. albicans and E. coli at non-antifungal pharmacological doses. In a secondary fungal infection model following CLP-induced sepsis, ClioQ (6 mg/kg) treatment significantly enhanced macrophage clearance efficiency against pathogens and markedly improved host survival. Conclusion ClioQ was identified as a potential immunomodulator that enhances host defense capabilities by directly repairing phagocytic dysfunction in macrophages during the immunosuppressive phase of sepsis. This approach significantly improves organ injury and increases survival rates, offering a novel candidate strategy for immune-adjuvant therapy in sepsis.

Superficial fungal infections are common worldwide and significantly impact public health. Understanding patients’ knowledge, attitudes, and practices (KAP) regarding their treatment and prognosis is essential for addressing gaps in care. This cross-sectional study utilized a self-designed KAP questionnaire to assess 456 patients with superficial fungal infections between October 15, 2023, and March 15, 2024. Data analysis included descriptive statistics, logistic regression, and structural equation modeling. The mean scores for knowledge, attitudes, and practices were 14.85 ± 7.75, 27.01 ± 4.28, and 22.91 ± 4.17, respectively. Key findings revealed that 32% of patients were unsure about long-term treatment, 51% misunderstood corticosteroid use, and 69% believed medications could be stopped once symptoms improved. Significant demographic disparities in KAP scores were observed, highlighting the influence of gender, age, education, and income. Patients exhibit inadequate knowledge, predominantly negative attitudes, and suboptimal practices regarding superficial fungal infections. Targeted educational interventions are crucial for enhancing treatment adherence, addressing misconceptions, and reducing antifungal resistance.

Ethnopharmacological relevance Dahuang Mudan decoction (DMD) is a traditional Chinese prescription from Zhang Zhongjing's Synopsis of the Golden Chamber. In clinical practice, it is often used in the treatment of infectious diseases. Aim of the study To assess the therapeutic effect of DMD and its disassembled prescriptions on septic mice, and explore its potential mechanism. Materials and methods Cecal ligation and puncture (CLP) sepsis and endotoxemia mice models were established. The effects of DMD, its disassembled prescriptions and active compounds were studied. Xuebijing injection (XBJ) was used as positive drug. Mice 7-day survival rates, blood biochemical markers, hematoxylin and eosin (HE) staining and immune cell infiltration were used to evaluate the overall protective effect of the drugs on mice. Inflammatory cytokines and coagulation activation indicators were detected by enzyme-linked immunosorbent assay (ELISA). Results DMD, its Huoxue prescription, constituent drugs Mudanpi (MDP) and Taoren (TR) significantly protected mice with sepsis, improved the survival rate, reduced the degree of organ damage, and reduced the infiltration of immune cells in the lung tissues. The protective effect is comparable to that of XBJ. MDP and TR inhibited the levels of inflammatory factors and coagulation activation in septic mice. Paeonol and paeoniflorin in MDP showed significant protective effects on septic mice, and inhibited inflammatory cytokines level and coagulation activation. Conclusion These results confirm that DMD and its disassembled prescriptions have good therapeutic effect on septic mice, and the mechanism may be related to inhibition of the inflammatory response and coagulation activation.

Rationale: The lungs are the most prevalent site for invasive fungal infections, and the diagnosis and treatment of pulmonary fungal infections (PFIs) pose significant challenges, accompanied by a substantial disease burden. Global factors will likely enhance the risk of PFIs in the future. Assessing the global burden of PFIs is crucial for implementing appropriate measures for prevention and control. Objectives: To evaluate the burden of PFIs at the global, regional, and national levels from 1990 to 2021 and make projections for 2044. Methods: Data on deaths and disability-adjusted life years due to PFIs were extracted from the Global Burden of Disease database. Linear regression, complex inequality measures, and the Nordpred model were used for analysis and visualization. Measurements and Main Results: In 2021, the global incidence of PFIs was estimated at 5.62 million cases (95% uncertainty interval [UI], 4.93 to 6.40 million), with 45,542 deaths (95% UI, 39,299 to 51,944). The age-standardized mortality rate was 0.56 per 100,000 (95% UI, 0.48 to 0.64 per 100,000). From 1990 to 2021, the estimated annual percentage change was -1.03% (95% confidence interval, -1.13% to -0.93%), with an observed increase in mortality rates in low- and middle-income countries. The mortality rate significantly increased among individuals aged ⩾50 years. By 2044, more than 87,000 deaths are expected from PFIs, at a rate of about 0.58 per 100,000. Conclusions: PFIs represent a significant global challenge that warrants attention and necessitates international collaboration to collectively address this issue.

Remimazolam (Rema) is a novel anesthetic that is widely used in anesthesia and sedation in critically ill patients. Notably, Rema exerts effects in patients through activation of the γ‑aminobutyric acid (GABA) receptor. GABA may alleviate myocardial ischemia/reperfusion (I/R) injury; however, the impact of Rema and underlying molecular mechanism in myocardial I/R injury remain to be fully understood. Therefore, the present study aimed to investigate the effects of Rema on cardiac I/R injury and to determine the underlying mechanisms. An acute myocardial I/R model was established by ligating the left anterior descending artery in adult male C57BL/6 mice (8‑10 weeks). Cultured Raw264.7 cells treated with lipopolysaccharide (LPS) were also used to investigate the effect of Rema on macrophages. The results of the present study revealed that Rema improved I/R‑induced cardiac dysfunction by increasing the ejection fraction value and reducing the myocardial infarction area. In addition, Rema also alleviated I/R‑induced cardiac inflammatory cell infiltration based on H&E and immunofluorescence staining. Transmission electron microscopy and ROS measurements showed that Rema improved I/R‑induced mitochondrial structural disruption and oxidative stress in cardiomyocytes. Transcriptomics analysis and reverse transcription‑quantitative PCR revealed that Rema alleviated I/R‑induced release of inflammatory factors and cytokines by inhibiting the expression of IL‑1β, IL‑6, C‑C chemokine receptor 2 and C‑X‑C motif chemokine ligand 5. Rema also inhibited I/R‑induced CD68+ cell proliferation, IL‑1β release, and NOD‑like receptor thermal protein domain associated protein 3 (NLRP3) and IL‑1β expression. The results of in vitro assays revealed that Rema inhibited LPS‑induced increases in IL‑1β, IL‑6 and TNF‑α expression and release in cultured RAW264.7 macrophages. In conclusion, the present study revealed that Rema may alleviate I/R‑induced cardiac dysfunction and myocardial injury by inhibiting oxidative stress and inflammatory responses via the NLRP3/IL‑1β pathway.

Background Sepsis-induced coagulopathy (SIC) is often a sign of high mortality and poor prognosis in patients with sepsis. Thrombomodulin (TM) plays an important anticoagulant role by activating protein (AP)C. Objectives Our previous study has shown that the overexpression of Nur77 upregulates TM expression in human umbilical vein endothelial cells (HUVECs). This study aimed to investigate whether upregulation of Nur77 using cytosporone (Csn)-B could ameliorate SIC. Methods A mouse model of SIC was prepared by cecum ligation and puncture (CLP) operation. Five hours after CLP, coagulation-related indicators and histopathologic injury of the liver, lungs, and kidneys were investigated. The effect of Csn-B on the clotting time of HUVECs transfected with Nur77 or TM small-interfering RNA in response to tumor necrosis factor α stimulation was observed. The effects of Csn-B on survival, organ damage, microthrombosis, coagulation factors, TM activated protein C anticoagulant system, fibrinolytic system, and complement system were observed in vascular endothelial conditional knockout Nur77 mice after CLP. Results Sepsis-induced upregulation of Nur77 in vascular endothelial cells. Knockout of Nur77 in vascular endothelium exacerbated organ damage and early coagulation dysfunction in sepsis. Csn-B attenuated the procoagulant response of HUVEC to tumor necrosis factor α stimulation, which is dependent on the activation of Nur77-TM pathway. Furthermore, Csn-B relied on activation of vascular endothelial Nur77 to inhibit the increase of coagulation factors, enhance activation of TM activated protein C, restore fibrinolysis homeostasis, and inhibit C3 and C5 activation to ameliorate hypercoagulability in SIC. Conclusion Csn-B improves early coagulopathy in sepsis by increasing endogenous TM through upregulating Nur77 in the vascular endothelium.

Messenger RNA (mRNA) vaccines have demonstrated significant potential in cancer immunotherapy by activating both innate and adaptive immunity. However, the detailed cellular and molecular dynamics underpinning these systemic immune responses remain incompletely understood. In this study, we characterized the systemic immune landscape following human papillomavirus (HPV)-targeted mRNA-lipid nanoparticle (LNP) vaccination using single-cell RNA sequencing (scRNA-seq) in a murine model of HPV-positive head and neck squamous cell carcinoma (HNSCC). Our study revealed a coordinated remodeling of the systemic immune landscape, involving the tumor microenvironment (TME), tumor-draining lymph nodes (TDLNs), spleen, and blood. Notably, we pioneered a distinct interferon-stimulated gene (ISG) signature across multiple lymphoid subsets in TDLNs, driven by the LNP component, which contributed to rapid, non-antigen-specific immune activation. Additionally, HPV mRNA-LNP vaccination induced an antigen-specific cycling burst of immune cells that mediated tumor control through a systemic coordination of multi-directional differentiation into anti-tumor cell compositions. These findings enhance our understanding of how mRNA-LNP vaccination orchestrates systemic anti-tumor responses and highlight the therapeutic potential of targeting ISG-expressing and cycling immune cells to improve vaccine efficacy, paving the way for future clinical applications in HPV-related cancers.

Edwardsiella piscicida is a highly pathogenic and stress-resistant bacterium that poses significant threats to the aquaculture industry. Ecotin (EcoT) is a multifunctional serine protease inhibitor that plays important roles in the pathogenicity of various bacteria. Its ability to inhibit host proteases and protect bacteria from immune attacks makes it a significant factor in bacterial infections. However, the role of EcoT in bacterial virulence remains largely unexplored. This study presents the first comprehensive elucidation of the diverse and critical roles of EcoT in E. piscicida. Our findings revealed that the mutation of ecoT significantly increases the production of bacterial extracellular polysaccharides (EPS) and enhances bacterial biofilm production. Under the strong acid stress, the expression of ecoT was significantly induced. Consistently, the mutation of ecoT significantly reduced bacterial capability to consume H+ ions and impaired its survival under strong acid stress conditions. Infection experiments demonstrated that ecoT deletion diminished the bacterium's resistance to host serum-mediated killing, proliferation within phagocytes, dissemination in immune tissues, and overall virulence. Based on these findings, we have formulated a hypothesis concerning the mechanism of action of EcoT. Under acidic conditions within the host, E. piscicida increases the levels of the periplasmic protein EcoT. EcoT enhances bacterial resistance to acidic stress, thereby facilitating bacterial survival within host cells. Concurrently, EcoT reduces EPS production and inhibits host protease activity, aiding the bacteria in evading the host's immune response. Our results suggest that EcoT functions as a novel virulence factor in E. piscicida, underscoring its significance in bacterial biofilm formation, stress tolerance, and pathogenicity.

Constant activation of stimulator of interferon genes (STING), resulting from aberrant metabolism or mutations in STING1, can initiate inflammatory damage or autoimmune disease. STING antagonists have the potential to be used as therapeutics for inflammatory and autoimmune diseases. Based on the structures of the covalent STING inhibitor H151 and C178, we designed, synthesized, and evaluated a novel series of indole derivatives for STING inhibition. Several compounds exhibited efficacious STING inhibitory activity. One of these novel chemical entities, 4dc, was more potent than H151, with IC50 values of 0.14 μM in RAW-LuciaTM ISG cells and 0.39 μM in THP1-Dual™ cells. The compound effectively relieved the symptoms of renal injury in a cisplatin-induced acute kidney injury mouse model. Compound 4dc represents a new chemotype of STING inhibitor that deserves further investigation as anti-inflammatory agent.

Background:Previous studies have demonstrated a significant association between neuroinflammation and major depressive disorder (MDD). (6aS,10S,11aR,11bR,11cS)-10-methylamino-dodecahydro-3a,7a-diaza-benzo(de)anthracene-8-thione (MASM), a derivative of matrine, has recently been shown to display anti-inflammatory properties. However, its effects on lipopolysaccharide (LPS)-induced depression and the underlying mechanisms remain unexplored. This study aimed to assess the effects of MASM on depressive-like behaviors induced by LPS and to investigate the potential mechanisms involved. Methods:Following intraperitoneal injection of LPS (0.83 mg/kg), MASM was administered. Depressive-like behaviors were assessed through the forced swim test (FST) and tail suspension test (TST). To further explore the mechanisms, LPS-induced BV2 microglial cell models were established. Enzyme-linked immunosorbent assay (ELISA) was used to quantify the expression of TNF-α and high mobility group box 1 (HMGB1), while immunoblotting was performed to assess heme oxygenase-1 (HO-1), sirtuin 1 (SIRT-1), p62, and microtubule-associated protein 1A/1B-light chain 3-phosphatidylethanolamine conjugate (LC3-II) expression. Reactive oxygen species (ROS) levels were evaluated using flow cytometry. Results:MASM pretreatment markedly ameliorated acute depressive-like behaviors in LPS-treated mice and upregulated HO-1 expression in the hippocampus. In LPS-stimulated BV2 cells, MASM reduced the levels of proinflammatory markers TNF-α and HMGB1. Furthermore, MASM mitigated LPS-induced oxidative stress, as evidenced by increased ATP, HO-1, and SIRT-1 levels, along with decreased ROS levels. MASM also restored autophagic function, demonstrated by increased LC3-II expression and reduced p62 levels. Conclusion:These findings suggests that MASM alleviates LPS-induced neuroinflammation and acute depressive-like behaviors, possibly by reducing oxidative stress and promoting autophagy.

A 43-year-old male patient presented with recurrent edema in different anatomical sites for over 10 years, with facial edema worsening 1 day prior to admission. He had been repeatedly admitted to dermatology, general surgery, and emergency departments of external hospitals due to " acute abdomen" and " laryngeal edema, " resistant to antihistamines and glucocorticoid therapy. Physical examination revealed non-pitting swelling of the right upper eyelid, bilateral cheeks, and lips asymmetrically. On the night of admission, he developed acute laryngeal edema with dyspnea, which was promptly treated, leading to clinical stabilization. Laboratory screening during the attack revealed decreased serum complement C4 levels, along with reduced functional activity and concentration of C1 esterase inhibitor, confirming a diagnosis of type 1 hereditary angioedema. The patient received lanadelumab for prophylaxis and achieved satisfactory clinical outcomes. He remains under long-term follow-up.

Toxoplasma gondii, an obligate intracellular protozoan parasite infecting nucleated cells of warm-blooded vertebrates, causes severe complications in immunocompromised hosts. Current therapies remain limited by suboptimal efficacy and toxicity, necessitating novel anti-toxoplasmic agents. Piceatannol (PIC), a natural stilbenoid, demonstrates multifaceted bioactivity including antimicrobial and anti-parasitic effects, suggesting therapeutic potential against T. gondii. Our previous study revealed PIC’s potent anti-parasitic activity, selectively inhibiting T. gondii proliferation and altering parasite morphology without host cytotoxicity. In this study, mechanistic analyses indicated that PIC disrupts mitochondrial integrity in tachyzoites, reducing mitochondrial membrane potential and ATP production while elevating ROS levels. Transcriptomic profiling identified significant suppression of oxidative phosphorylation-related genes, consistent with mitochondrial dysfunction. These findings establish PIC as a promising candidate targeting T. gondii through the mechanism of mitochondrial impairment.

Tumor-infiltrating CD4+ T cells orchestrate the adaptive immune response through remarkable plasticity, and the expression patterns of exhaustion-related inhibitory receptors in these cells differ significantly from those of CD8+ T cells. Thus, a better understanding of the molecular basis of CD4+ T cell exhaustion and their responses to immune checkpoint blockade (ICB) is required. Here, we integrated multiomics approaches to define the phenotypic and molecular profiles of exhausted CD4+ T cells in oropharyngeal squamous cell carcinoma (OPSCC). Two distinct immune-promoting (Module 1) and immunosuppressive (Module 2) functional modules in tumor-infiltrating CD4+ T cells were identified, and both the immune-promoting function of Module 1 cells and immunosuppressive function of Module 2 cells were positively associated with their corresponding exhaustion states. Furthermore, the application of ICBs targeting effector CD4+ T cells in Module 1 (αPD-1) and Treg cells in Module 2 (αCTLA-4) in mouse models could help reinvigorate the effector function of Module 1-exhausted CD4+ T cells and reduce the immunosuppressive function of Module 2-exhausted CD4+ T cells, ultimately promoting OPSCC tumor regression. Taken together, our study provides a crucial cellular basis for the selection of optimal ICB in treating OPSCC.

Nonalcoholic fatty liver disease (NAFLD) is the most prevalent type of chronic liver disease. However, the disease is underappreciated as a remarkable chronic disorder as there are rare managing strategies. Several studies have focused on determining NAFLD-caused hepatocyte death to elucidate the disease pathoetiology and suggest functional therapeutic and diagnostic options. Pyroptosis, ferroptosis, and necroptosis are the main subtypes of non-apoptotic regulated cell deaths (RCDs), each of which represents particular characteristics. Considering the complexity of the findings, the present study aimed to review these types of RCDs and their contribution to NAFLD progression, and subsequently discuss in detail the role of necroptosis in the pathoetiology, diagnosis, and treatment of the disease. The study revealed that necroptosis is involved in the occurrence of NAFLD and its progression towards steatohepatitis and cancer, hence it has potential in diagnostic and therapeutic approaches. Nevertheless, further studies are necessary.