Tuberculosis is a top killer among infectious diseases. Traditional tuberculosis treatment drugs have serious resistance issues and long treatment courses. Therefore, continuous improvement and new treatment strategies against TB are urgently required. Pyrvinium pamoate is a Food and Drug Administration (FDA) approved anthelminthic drug. Recently we have reported that pyrvinium pamoate can decrease mycobacterial burdens in Mycobacterium tuberculosis (M. tuberculosis)-infected macrophages and mice. However, the mechanism by which pyrvinium pamoate inhibits the survival of intracellular M. tuberculosis remains to be explored. In this study, we discovered that low dose (0.2 μg/mL, less than IC50, half-maximal inhibitory concentration) of pyrvinium pamoate could inhibit the survival of intracellular M. tuberculosis H37Rv/H37Ra growth through suppressing ferroptosis of the infected macrophage. We found that pyrvinium pamoate could bind to casein kinase (CK)1α protein and suppress M. tuberculosis- or RSL3 (a well-known inducer of cell ferroptosis)-induced lipid peroxidation ferroptosis in macrophages through reducing ATF4-xCT-GSH-GPX4 expression and activation of YAP1-ACSL4 and TFRC-Fe3+ pathways. CK1α siRNA or its inhibitor D4476 can reverse above effects by pyrvinium pamoate on the ferroptosis and intracellular M. tuberculosis survival. We unveil a previously unrecognized and multifaceted mechanism by which pyrvinium pamoate, via targeting CK1α, inhibits M. tuberculosis-induced ferroptosis. We propose that pyrvinium pamoate holds great promise as a host-directed therapy (HDT) drug for mycobacterial-induced ferroptosis.

BACKGROUND/OBJECTIVES:The mycobacterial complex cell envelope serves as a formidable barrier against host immunity and antibiotics. Lipomannan (LM) and lipoarabinomannan (LAM) are key structural components of the mycobacterial envelope and potent immunomodulators. The mycobacterial lipoarabinomannan biosynthesis mannosyltransferase MptC modifies the multiple α-(1→2)-linked branched mannan residues of LAM in the mycobacteria. However, the role of MptC in mycobacterial infectivity, antibiotic susceptibility and host immune regulation remains poorly understood. METHODS:An mptC (also named MSMEG_4247) knockout Mycobacterium smegmatis mc2-155 (M. smeg) strain (designated as M. smegΔmptC) was generated using CRISPR-Cas12a technology. The effects of MptC on bacterial physiology, cell wall permeability, drug sensitivity, immune cell function, and survival during infection are analyzed through glycogen staining, drug sensitivity tests, and cellular and mouse infection models. RESULTS:MptC deficiency results in a loss of LM and increase in LAM synthesis. The M. smegΔmptC mutant strain exhibits enhanced cell wall permeability and reduces hydrophobicity. Functionally, the mptC knockout strain increases the intracellular cytokines (IFN-γ, TNF-a and IL-17) production of T cells in mice. Consequently, results based on both macrophage and mouse infection models demonstrate that the M. smegΔmptC strain has less bacterial loads and higher susceptibility to antibiotic rifampicin. CONCLUSIONS:Mannosyltransferase MptC plays an important role in maintaining cell wall integrity (via LM/LAM synthesis), regulating T cell responses, and influencing antibiotic susceptibility in mycobacteria.

Tuberculosis (TB) remains a major global health burden, and Bacille Calmette-Guérin (BCG) provides inconsistent protection against adult pulmonary disease. Adjuvant strategies to enhance BCG efficacy are urgently needed. Here we show that whole β-glucan particles (WGP) enhance BCG-induced immune responses and improve protection against mycobacterial challenge. In vitro, WGP enhances macrophage functions associated with antigen processing during BCG exposure, including phagocytosis, lysosomal acidification, intracellular degradation, and the upregulation of MHC-II/CD80/CD86. WGP co-exposure partially attenuates the early IL-10 response induced by BCG. Mechanistically, whole β-glucan particles activate the Dectin-1-JAK1-STAT1 pathway, and disruption of this axis reduces MHC-II upregulation and impairs macrophage-supported OT-II CD4+ T-cell proliferation and IFN-γ production. In vivo, using female wild-type and Dectin-1 knockout C57BL/6 J mice, we find that split-site administration of WGP and BCG is better tolerated than same-site mixing and is associated with a less inflammatory early pulmonary myeloid profile. Under this optimized regimen, WGP plus BCG enhances CD4+ memory-associated responses and improves bacterial control while reducing lung inflammation in a Mycobacterium tuberculosis H37Ra challenge model. These findings support further evaluation of WGP as an adjunct candidate for BCG-based vaccination strategies and may inform the development of improved tuberculosis vaccines.

The 2025 National Natural Science Foundation of China (NSFC) Major Research Program “Decoding the Glycan Code of Life” has been sucessfully held at the East Lake International Conference Center in Wuhan, China, from December 17–19, 2025. The successful convening of this annual exchange meeting not only showcased significant progress in glycan synthesis and detection technologies, functional and regulatory mechanisms, and translational applications, but also strengthened interdisciplinary collaboration networks, sharpened core scientific objectives, and laid a solid foundation for China to maintain a leading international position in this strategic frontier field.

Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis (TB), is the leading cause of infectious disease-related death. As a major intracellular pathogen, Mtb can escape clearance by the immune system, but the underlying molecular mechanisms remain incompletely elucidated. Specific genomic regions of deletion (RD)-encoded proteins in virulent Mtb H37Rv have been implicated in modulating pathogenicity and immunity. Here, we report a novel RD15-encoding protein, Rv1977 (a mycobacterial cell wall protein with a size of 39 kDa, named MEM39), which facilitates Mtb survival in macrophages. The survival of the Mtb H37Rv MEM39-deficient strain is reduced in both macrophage and murine infection models. Furthermore, the mycobacterial MEM39 protein binds fructose-diphosphate aldolase A (ALDOA), a key enzyme of glycolysis, thereby impairing ALDOA enzyme activity, disrupting macrophage metabolite flux, and reducing lactate production. The MEM39-ALDOA interaction also suppresses lysosomal acidification; reduces NLRP3 inflammasome activation and the production of proinflammatory cytokines (TNF-α, IL-6 and IL-1β); and thereby promotes bacterial survival within macrophages. Disruption of the interaction between MEM39-ALDOA and a cell-penetrating synthetic peptide (VLARYASICQ) significantly suppressed Mtb survival by restoring lactate production, lysosome acidification and proinflammatory cytokine production in both macrophage and mouse infection models. These findings revealed that mycobacterial MEM39 negatively regulates host immune defense through reprogramming ALDOA-mediated glycolysis in macrophages, thereby forming a "mycobacterial MEM39 virulence factor-glycolysis metabolism-immunity" regulatory axis. Targeting MEM39 or the MEM39-ALDOA interaction interface holds promise as a new therapeutic strategy against tuberculosis.

Introduction Pyroptosis is an important inflammatory form of cell death and Mycobacterium tuberculosis ( M.tb ) chronic infection triggers excessive inflammatory pyroptosis of macrophages. Our previous research has confirmed that a small compound pyrvinium pamoate (PP) could inhibit inflammatory pathological changes and mycobacterial burden in M.tb -infected mice, but the potential mechanism of PP for inhibiting M.tb -induced inflammation remains unexplored. Methods The effects of PP on the NLRP3-ASC-Casp1 inflammasome assembly and activation, gasdermin D (GSDMD) mediated pyroptosis and inflammatory cytokines expression were assessed in human THP-1-derived macrophages after M.tb H37Rv/H37Ra/ Salmonella typhimurium ( S. typhimurium ) infection or LPS treatment by Transcriptome sequencing, RT-qPCR, Co-immunoprecipitation and Western Blot (WB) analysis. The lactate dehydrogenase (LDH) release assay was used to evaluate the CC50 of PP in M.tb -infected THP-1 cells. Results We found that M.tb / S. typhimurium infection and LPS treatment significantly activate NLRP3-ASC-Casp1 inflammasome activation, GSDMD-mediated pyroptosis and inflammatory cytokines (IL-1β and IL-18) expression in macrophages, whereas PP could suppress these inflammatory effects in a dose dependent manner. Regarding the PP-inhibition mechanism, we further found that this inhibitory activity is mediated through the PP-targeting casein kinase 1A1 (CK1α)–β-catenin–NF-κB pathway and CK1α–NRF2–mitochondrial oxidative phosphorylation (OXPHOS) pathway. In addition, a CK1α specific inhibitor D4476 or CK1α siRNA could reverse these inhibitory effects of PP on bacteria-induced inflammatory responses in macrophages. Conclusions This study reveals a previously unreported mechanism that pyrvinium can inhibit NLRP3 inflammasome and GSDMD–IL-1β inflammatory pyroptosis via targeting suppressing CK1α–β-catenin–NF-κB and CK1α–NRF2–mitochondrial OXPHOS pathways, suggesting that pyrvinium pamoate holds great promise as a host directed therapy (HDT) drug for mycobacterial-induced excessive inflammatory response.

Tuberculosis (TB) is one of the ten major factors threatening human life and health. At present, many factors limit the application of existing anti-tuberculosis drugs, such as a small range of available drug options, poor treatment compliance, and severe toxic and side effects. It is extremely urgent to develop novel anti-tuberculosis drugs. DprE1 is a potential anti-mycobacterial cell wall target, and some DprE1 inhibitors have entered the clinical research stage. Our research group found DprE1 inhibitor G50 with similar activity as isoniazid through virtual screening in the early stage. To obtain better DprE1 inhibitors, 45 new compounds were designed and synthesized based on the structure of G50. Among them, 12 selected DprE1 enzyme inhibitors could significantly inhibit the growth of Mycobacterium tuberculosis (M.tb) H37Ra and H37Rv growth in vitro. The MIC50 value of compound 42 against M.tb H37Ra is 1.071 ± 0.041 μM, with the selective index (SI) value of 186.74 (the SI value of linezolid is 119.9). Compared to G50, compound 42 exhibits a 5-fold increase in DprE1 enzyme inhibitory activity. In addition, the binding affinity of compound 42 is equivalent to that of G50. This study further enriches the examples of developing DprE1 inhibitors based on the backbone of pyrimidinetrione and also provides potential anti-tuberculosis lead compounds.

RNA viruses have high mutation rates and constitute an increasing global risk. As the viral target approach to develop antiviral drugs is inadequate for responding to an increasing diversity of viruses, an urgent need exists for the development of new antivirals to prevent future outbreaks. Here, we show that aglycone ionophores maduramycin (Mad) and endusamycin (End) from Streptomyces are broadly virucidal against cytoplasmic replicated viruses, including Japanese encephalitis virus (JEV), rabies virus, hepatitis C virus, vesicular stomatitis virus, hantavirus, dengue virus, Zika virus, chikungunya virus, and SARS-CoV-2 in vitro. Mechanistic studies suggest Mad and End can target actin filaments and displace the DNase-I-binding loop (D-loop) into an outward conformation for stabilizing actin filaments and primarily inhibit viral replication. Liposome-encapsulated Mad or End fully protects mice against JEV infection in vivo. Thus, our results may provide potential and naturally produced antivirals to prevent the spread of viruses in animals.

BACKGROUND:Environmental pollutants have been found to contribute to the development and acute exacerbation of asthma. Microplastics (MPs) have received widespread attention as an emerging global pollutant. Airborne MPs can cause various adverse health effects. Due to their hydrophobicity, MPs can act as a carrier for other pollutants, pathogens, and allergens. This carrier effect of MPs may adsorb allergens and thus make the body exposed to MPs and a large number of allergens simultaneously. We hypothesized that co-exposure to inhaled MPs and aeroallergens may promote the development of airway inflammation of asthma by disrupting the airway epithelial barrier. METHODS:The effects of co-exposure to Polyethylene microplastics (PE-MPs) and allergens on allergic airway inflammation and airway epithelial barrier were examined in a mouse model of asthma. The mice were divided into four groups: (i) Control group, treated only with PBS; (ii) MP group, exposed to PE-MPs and PBS; (iii) HDM group, mice were sensitized and challenged with HDM, and intranasally treated with PBS; (iv) HDM + MP group, mice were sensitized and challenged with HDM, and intranasally treated with PE-MPs. Histology and ELISA assays were used to evaluate the severity of airway inflammation. FITC-dextran permeability assay, immunofluorescence assay, and RT-PCR were used to evaluate the airway epithelial barrier function and the expression of relevant molecules. Transcriptomics analysis with lung tissue sequencing was conducted to identify possible pathways responsible for the effects of PE-MPs. RESULTS:Co-exposure of mice to PE-MPs and HDM induced a higher degree of inflammatory cell infiltration, bronchial goblet cell hyperplasia, collagen deposition, allergen sensitization, and Th2 immune bias than exposure to HDM alone. Co-exposure to PE-MPs and HDM aggravated oxidative stress injury in the lung and the production of cytokine IL-33 in the BALF. In addition, co-exposure of mice to PE-MPs and HDM resulted in a more pronounced decrease in the expression of relevant molecules of the airway epithelial barrier and more significant increase in the permeability of airway epithelia. Lung tissue transcriptomics analysis revealed that PE-MPs exposure was associated with CXCL1 signaling and neutrophil activation. CONCLUSION:Co-exposure to MPs and HDM may promote airway inflammation and airway epithelial barrier disruption and induce immune responses characterized by CXCL1 signaling and neutrophilic inflammation.

Hepatitis B virus (HBV) infection is a major etiological factor in the development of hepatocellular carcinoma (HCC). Despite extensive research efforts, the precise molecular mechanisms and critical host factors driving HBV-induced epithelial-mesenchymal transition (EMT), liver fibrosis and hepatocarcinogenesis remain to be explored. Emerging evidence has identified aberrant expression of Troponin T1 (TNNT1) in malignancies, implicating its potential role in HCC progression. However, the specific role and mechanism of TNNT1 in HBV-associated HCC remain elusive. In this study, we demonstrate that TNNT1 expression is markedly upregulated in HBV-positive HCC tissues, HBV infection/replication cell models and AAV-HBV1.3-infected mouse models. Mechanistically, HBV activates the transcription factor c-Myc via the PI3K/AKT/mTOR signaling pathway. HBV promotes HCC cell proliferation and EMT markers (Cyclin D1, Vimentin increased, E-cadherin decreased) and liver fibrosis marker α-smooth muscle actin (α-SMA) expression in a TNNT1-dependent manner. HBV infection-induced EMT and liver fibrosis can be abolished by hepatic-specific TNNT1 knockout or knockdown in mice. These findings provide novel insights into the role of TNNT1 in HBV-driven EMT and liver fibrosis, and establish a foundation for further exploration of TNNT1 as a potential therapeutic target in HBV-associated HCC progress.

BACKGROUND:Hepatocellular carcinoma (HCC) is a major global health issue, with poor prognosis often associated with dysregulated metabolic pathways, especially lactate metabolism. This study explored the prognostic significance of lactate-associated genes in HCC and their potential as therapeutic targets. METHODS:We analyzed RNA-seq and clinical data from 374 patients with HCC from The Cancer Genome Atlas (TCGA) database. Using Cox regression, LASSO analysis, and Kaplan-Meier survival curves, we identified key lactate-associated genes associated with patient outcomes. Functional validations, including Western blot, flow cytometry, and molecular docking studies, were performed to confirm the biological impact of these genes. RESULTS:G6PD, IK, and CALML5 were identified as significant prognostic markers for HCC. A prognostic model was developed that effectively stratified patients into risk groups, which correlated with survival. G6PD's role in immune modulation and its potential as a drug target were validated through biochemical assays and computational analyses. Functional assays in HepG2 cells confirmed that alterations in G6PD expression affect T cell activity, with knockdown enhancing IFN-γ production and overexpression inhibiting it, demonstrating G6PD's role in immune evasion. CONCLUSIONS:This study establishes lactate metabolism genes, particularly G6PD, as key prognostic markers in HCC. The validation of G6PD's immunomodulatory effects further supports its potential as a therapeutic target for strategies aimed at enhancing immune surveillance and treatment outcomes in HCC.

PURPOSE:The rising prevalence of food allergy (FA) has prompted investigations into dietary factors such as trans fatty acids (TFAs). While ruminant TFAs may protect against allergies, the role of industrial TFAs remains unclear. This study evaluated the effects of industrial TFAs on FA in a murine model. METHODS:20 Balb/c mice were divided into 4 groups: control (standard diet), OVA (ovalbumin-sensitized, standard diet), TFAs (industrial TFA-enriched diet), and TFAs+OVA (TFA diet + OVA sensitization). After two weeks, OVA and TFAs+OVA groups underwent OVA sensitization/challenge. Symptoms (anal temperature drop, diarrhea), serum immunoglobulin E (tIgE, OVA-sIgE), cytokines, immune cell profiles, and gut microbiota were analyzed. RESULTS:The incidence rates of anal temperature drop and diarrhea, the serum levels of tIgE, OVA-sIgE, interleukin-4 were significantly higher in the OVA and TFAs+OVA groups compared to the control group. The TFAs+OVA group had a higher degree of anal temperature drop and diarrhea score, and higher serum levels of tIgE and OVA-sIgE compared to the OVA group. The expression of interferon-γ mRNA and the numbers of Th1 cells increased in the spleen of the TFAs and TFAs+OVA group compared to the control group, whereas the numbers of spleen Th2 cells were significantly elevated in the TFAs, OVA, and TFAs+OVA groups compared to the control group. In addition, the numbers of mast cells (MCs) in the esophagus and intestinal mucosa, and the serum concentrations of MCs protease-1 were significantly increased in TFAs, OVA, and TFAs+OVA groups compared to the control group. Cecal microflora among these groups exhibited distinct patterns of differential diversity and composition. CONCLUSIONS:Industrial TFAs may promote OVA-induced FA, Th1 and Th2-associated inflammation in mouse model, accompanied by the activation of MCs and intestinal microbiome dysbiosis.

BACKGROUND:Type 2 inflammation has emerged as a pivotal mechanism for asthma, which involves both innate and adaptive immunity. Human ficolin (FCN)-2 (L-ficolin, P35) and its mouse homolog FCN-A are one of the major pattern recognition molecules of plasma/serum, acting as important initiators of the lectin complement system and playing important roles in immunity, including respiratory immunity. However, little is known about the role of FCN-2/A in allergic asthma. METHODS:Serum FCN-2 and IgE levels in 90 allergic asthmatic patients and 48 healthy controls were measured by ELISA. Aeroallergen house dust mite (HDM)-induced mouse model of asthma was generated in both wild type (WT) and FCN-A knockout (KO) mice. Mouse serum and bronchoalveolar lavage fluid (BALF) IgE levels, lung innate lymphoid cells (ILC)1/2/3, the expression of transcription factors GATA3, T-bet, and RORγt, and the concentrations of type 2 cytokines in serum and BALF were measured by FCM, RT-qPCR, Western blot, and ELISA. RESULTS:Serum FCN-2 concentrations in patients with allergic asthma were significantly lower than those in healthy controls. Similarly, lower serum and BALF FCN-A concentrations were observed in HDM-induced asthma mouse models compared to those of uninduced mice. In the asthma mouse model, FCN-A KO asthmatic mice had higher levels of total IgE and HDM-specific IgE (sIgE), β-hexosaminidase (β-HEX) and histamine secretion, as well as increased airway epithelial permeability with the release of FITC-dextran in sera, inflammatory cell infiltration and eosinophil counts, and displayed more severe disease symptoms with histological damage compared to WT asthmatic mice. FCN-A KO asthmatic mice showed decreased T-bet+ ILC1 and increased IL-5+/IL-13+ ILC2/ILC2 proportions, p-GATA3 expression, serum and BALF type 2 cytokines IL-4, IL-5, and IL-13, Th17 cytokine IL-17, and chemokines CCL2/4 production. Importantly, the administration of exogenous FCN-A protected against mouse allergic airway inflammation with decreased ILC2 proportions and type 2 cytokines expression, serum total and allergen-specific IgE production. These results suggest that FCN-A suppresses both ILC2 innate immunity and IgE-mediated adaptive immunity during asthma. CONCLUSION:Our findings provide previously unreported evidence that FCN-A protects against allergic asthma by suppressing lung ILC2-driven type 2 inflammation.

Introduction:The fat mass and obesity-associated protein (FTO), a key RNA N6-methyladenosine (m6A) demethylase, has been highlighted for its important role in inflammatory response. Emerging evidences link the O-GlcNAcylation to numerous human diseases, particularly inflammation. However, the specific role and underlying mechanism of FTO O-GlcNAcylation in inflammation remain elusive. Methods:The FTO O-GlcNAcylation modification was determined by co-immunoprecipitation (Co-IP) assay, metabolic glycan labeling combined with click reaction, and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Chromatin immunoprecipitation (ChIP)-qPCR and dual-luciferase reporter assay were used to determine FOXO1 binding to the Gfat2 promoter and the Gfat2 promoter activity during LPS stimulation. The FTO ubiquitination modification and the interaction between FTO and TRIM21 were detected by confocal microscopy, pull-down, and mass spectrometry analysis. The effects of FTO O-GlcNAcylation on the ubiquitination degradation of FTO were assessed by Co-IP and protein stability assays. The Socs1 mRNA m6A methylation levels were detected by m6A-RNA-immunoprecipitation (RIP)-qPCR. The myeloid-specific Fto deletion (myeFto -/-) mice, the macrophage depletion and reconstitution experiments, and enzyme-linked immunosorbent assays (ELISA) were used to evaluate inflammatory responses in Salmonella Typhimurium (S. Typhimurium) or bacterial endotoxin (lipopolysaccharide, LPS) induced sepsis mouse models. Results:We demonstrate that FTO undergoes O-GlcNAcylation specifically at the serine 95 (Ser95) site. LPS enhances FTO O-GlcNAcylation modification levels by increasing the FOXO1-regulated GFAT2 expression. O-GlcNAcylation of FTO promotes TRIM21-mediated K48-ubiquitination degradation of FTO, which further induces suppressor of cytokine signaling (Socs) 1 m6A methylation, thus sustains SOCS1 protein expression and suppresses multiple inflammatory cytokines IL-1β/IL-6/TNF-α production in LPS-stimulated macrophages. FTO O-GlcNAcylation mutation (S95A) aggravates S. Typhimurium or LPS-induced sepsis, while FTO O-GlcNAcylation suppresses the hyperinflammatory phenotype in mice. Promotion of O-GlcNAcylation by an OGA inhibitor Thiamet-G alleviates LPS-induced inflammatory responses and septic shock in mice. Conclusion:These findings reveal a mechanism that FTO O-GlcNAcylation promotes its ubiquitination degradation, and thus induces Socs1 m6A methylation and downregulates LPS-mediated inflammatory response, which maintains the negative feedback control of macrophage inflammatory cytokine storm in sepsis. Regulation of FTO O-GlcNAcylation may offer a potential therapeutic strategy for combating endotoxin-induced inflammatory disease and other FTO abnormal expression-associated diseases.

Objective Chemotherapy is one of the important therapeutic approaches for cancer treatment. However, the emergence of multidrug resistance and side effects significantly limit its application. To address these challenges, chemotherapy is often combined with other drugs or therapies. Among the 13 human fucosyltransferases (FUTs) identified, FUT8 (alpha-(1,6)-fucosyltransferase) is the only enzyme responsible for core fucosylation. Core fucosylation plays an important role in cancer occurrence, metastasis and chemotherapy resistance, making the suppression of FUT8 a potential strategy for reversing multidrug resistance. This study aims to evaluate the feasibility of combining the small molecule FUT8 inhibitor 2FF (2-deoxy-2-fluoro-L-fucose) with the clinical chemotherapeutic drug doxorubicin (DOX) for treating malignant tumors. Methods The human hepatocellular carcinoma cell line HepG2 and mouse colon cancer cell line CT26 cells were treated with 2FF, DOX or their combination and core fucosylation levels were assessed using Lectin blot. HepG2 and CT26 cells were exposed to 50 mu mol/L 2FF for 72 h, followed by treatment with a gradient concentration of DOX for 24 h. Cell viability and IC50 values were determined via the CCK-8 assay. Transwell invasion assays were conducted to evaluate the combined effect of 2FF and DOX on the invasion ability of HepG2 cells. Flow cytometry was performed to analyze the impact of 2FF, DOX and their combination on membrane PD-L1 expression of HepG2 cells. To assess the in vivo effect, 6- to 8-week-old female BALB/c mice (20-25 g), were subcutaneously injected with 1x10(6) CT26 cells into the right axilla (four groups, six mice in each group). After the average tumor volume reached 100 mm(3), mice were treated with DOX, 2FF, their combination, or saline (mock group) every other day. DOX was administrated intraperitoneally (2 mg/kg), 2FF intravenously (5 mg/kg), and the combination group, received the both treatment. Tumor size was measured every other day using a vernier caliper. Results This study demonstrated that DOX upregulates the core fucosylation levels in HepG2 and CT26 cells, while 2FF effectively inhibits this DOX-induced effect. Furthermone, 2FF enhanced the sensitivity of HepG2 and CT26 cells to DOX. The combination of 2FF and DOX synergistically inhibited the invasion ability of HepG2 cells, and enhanced the anti-tumor efficacy of CT26 subcutaneous tumor model in BALB/c mice. However the combination treatment led to weight loss in mice. In addition, DOX increased the cell surface PD-L1 expression in HepG2 cells, which was effectively suppressed by 2FF. Conclusion The FUT8 inhibitor 2FF effectively suppresses DOX-induced upregulation of core fucosylation and PD-L1 levels in tumor cells, and 2FF synergistically enhances the anticancer efficacy of DOX.

Accumulating evidence indicates that G-quadruplexes (G4s) are involved in transcriptional regulation. Previous studies have demonstrated that DHX36 preferentially resolves G4s, suggesting its potential impact on gene transcription mediated by these structures. However, systematic validation is required to establish a link between DHX36 activity and its roles in transcriptional regulation. In this study, we investigate the role of DHX36 in transcription. First, we employ the cleavage under targets and tagmentation (CUT&Tag), an efficient method for mapping protein–DNA interactions, to identify the binding sites in the chromatin of MCF-7 cells. Subsequently, we use the auxin-inducible degron (AID) protein degradation system and improved nascent RNA sequencing method acrylonitrile-mediated uridine-to-cytidine conversion sequencing (AMUC-seq) to pinpoint genes directly regulated by DHX36. Our results reveal a significant enrichment of G4 structures at DHX36 target sites, predominantly located in active genomic regions. In vitro assays further demonstrate DHX36's interaction with G4 sequences from three specific oncogenes. These findings underscore the potential role of DHX36 in modulating gene transcription through G4 structures.

Tuberculosis (TB), caused by Mycobacterium tuberculosis ( M. tb), remains one of the leading causes of fatal infectious diseases worldwide. The only licensed vaccine, Mycobacterium bovis Bacillus Calmette-Guérin (BCG), has variable efficacy against TB in adults. Insufficiency of immune cell function diminishes the protective effects of the BCG vaccine. It is critical to clarify the mechanism underlying the antimycobacterial immune response during BCG vaccination. Macrophage mannose receptor (MR) is important for enhancing the uptake and processing of glycoconjugated antigens from pathogens for presentation to T cells, but the roles of macrophage MR in the BCG-induced immune response against M. tb are not yet clear. Here, we discover that macrophage MR deficiency impairs the antimycobacterial immune response in BCG-vaccinated mice. Mechanistically, macrophage MR triggers JAK-STAT1 signaling, which promotes antigen presentation via upregulated MHC-II and induces IL-12 production by macrophages, contributing to CD4 + T cell activation and IFN-γ production. MR deficiency in macrophages reduces the vaccine efficacy of BCG and increases susceptibility to M. tb H37Ra challenge in mice. Our results suggest that MR is critical for macrophage antigen presentation and the antimycobacterial immune response to BCG vaccination and offer valuable guidance for the preventive strategy of BCG immunization.

BACKGROUND:Airway epithelial barrier dysfunction has been proved to contribute to the development of type 2 inflammation of asthma. Interleukin (IL)-37 is a negative regulator of immune responses and allergic airway inflammation. However, whether IL-37 has any effect on airway epithelial barrier has been unknown. METHODS:We evaluated the role of IL-37 in both mouse model and cultured 16HBE cells. Histology and ELISA assays were used to evaluate airway inflammation. FITC-dextran permeability assay was used to evaluate the airway epithelial barrier function. Immunofluorescence, western blot and quantitative Real-Time PCR (RT-PCR) were used to evaluate the distribution and expression of tight junction proteins. RT-PCR and Ca2+ fluorescence measurement were used to evaluate the mRNA expression and activity of store-operated calcium entry (SOCE). RESULTS:IL-37 inhibited house dust mite (HDM)-induced airway inflammation and decreased the levels of IgE in serum and type 2 cytokines in bronchoalveolar lavage fluid (BALF) compared to asthmatic mice. IL-37 protected against HDM-induced airway epithelial barrier dysfunction, including reduced leakage of FITC-dextran, enhanced expression of TJ proteins, and restored the membrane distribution of TJ proteins. Moreover, IL-37 decreased the level of IL-33 in the BALF of asthmatic mice and the supernatants of HDM-treated 16HBE cells. IL-37 decreased the peak level of Ca2+ fluorescence induced by thapsigargin and HDM, and inhibited the mRNA expression of Orai1, suggesting an inhibiting effect of IL-37 on SOCE in airway epithelial cells. CONCLUSION:IL-37 plays a protective role in airway inflammation and HDM-induced airway epithelial barrier dysfunction by inhibiting SOCE.

Hepatitis C virus (HCV) is a positive-stranded RNA virus that mainly causes chronic hepatitis, cirrhosis and hepatocellular carcinoma. Recently we confirmed m5C modifications within NS5A gene of HCV RNA genome. However, the roles of the m5C modification and its interaction with host proteins in regulating HCV’s life cycle, remain unexplored. Here, we demonstrate that HCV infection enhances the expression of the host m5C reader YBX1 through the transcription factor MAX. YBX1 acts as an m5C reader, recognizing the m5C-modified NS5A C7525 site in the HCV RNA genome and significantly enhancing HCV RNA stability. This m5C-modification is also required for YBX1 colocalization with lipid droplets and HCV Core protein. Moreover, YBX1 facilitates HCV RNA replication, as well as viral assembly/budding. The tryptophan residue at position 65 (W65) of YBX1 is critical for these functions. Knockout of YBX1 or the application of YBX1 inhibitor SU056 suppresses HCV RNA replication and viral protein translation. To our knowledge, this is the first report demonstrating that the interaction between host m5C reader YBX1 and HCV RNA m5C methylation facilitates viral replication. Therefore, hepatic-YBX1 knockdown holds promise as a potential host-directed strategy for HCV therapy.

Lung granuloma is a very common lung disease, and its specific diagnosis is important for determining the exact cause of the disease as well as the prognosis of the patient. And, an effective lung granuloma detection model based on computer-aided diagnostics (CAD) can help pathologists to localize granulomas, thereby improving the efficiency of the specific diagnosis. However, for lung granuloma detection models based on CAD, the significant size differences between granulomas and how to better utilize the morphological features of granulomas are both critical challenges to be addressed. In this paper, we propose an automatic method CRDet to localize granulomas in histopathological images and deal with these challenges. We first introduce the multi-scale feature extraction network with self-attention to extract features at different scales at the same time. Then, the features will be converted to circle representations of granulomas by circle representation detection heads to achieve the alignment of features and ground truth. In this way, we can also more effectively use the circular morphological features of granulomas. Finally, we propose a center point calibration method at the inference stage to further optimize the circle representation. For model evaluation, we built a lung granuloma circle representation dataset named LGCR, including 288 images from 50 subjects. Our method yielded 0.316 mAP and 0.571 mAR, outperforming the state-of-the-art object detection methods on our proposed LGCR.