The tumour microenvironment (TME) of relapsed/refractory (R/R) diffuse large B-cell lymphoma (DLBCL) patients is associated with resistance of DLBCL cells to CD19 CAR-T cells. How to improve TME in DLBCL and improve the efficacy of CAR-T cell therapy remains to be further explored. We observed the sensitivity of HBL-1/U2932 cells pretreated with BTK inhibitors (BTKi) to CAR-T cells with flow cytometry (FCM). The effect of pretreatment of BTKi on the polarisation state of alternative activated M2 macrophages was observed with FCM, real-time PCR and Western blot method. The effect of Notch1 agonist on expressions of Arg-1 protein, iNOS protein, Notch1 protein and RBP-J protein in alternative activated M2 macrophages was observed by Western blot method. Then the expression consistency of Notch-1 and RBP-J in activated M2 macrophages was observed by siRNA transfection of Notch-1. The cytotoxicity of CD19 CAR-T cells on HBL-1/U2932 cells pretreated with ibrutinib/orelabrutinib was higher than that of HBL-1/U2932 cells unpretreated with ibrutinib/orelabrutinib. Cytotoxicity of CAR-T cells to HBL-1 cells in coculture system with alternative activated M2 macrophages was very low. This drug resistance could be reversed by replacing the M2 macrophages (M2 macrophages after 48 h pretreatment with BTKi) in coculture system. Pretreatment with BTKi could down-regulate the expression of CD206 and IL-10 in activated M2 macrophages. And pretreatment with BTKi down-regulated the expression of Arg-1 and upregulated the expression of iNOS in activated M2 macrophages. The upregulation polarisation of M2 macrophages by Notch1 agonist could be reversed by BTKi. Expression of RBP-J protein decreased in alternative activated M2 macrophages by siRNA silencing Notch 1. Pretreatment with BTKi could down-regulate the polarisation of M2 macrophages and reverse the resistance of DLBCL cells which were cocultured with alternative activated M2 macrophages to CAR-T cells. This effect might be achieved by downregulating the Notch-RBP-J pathway.

WHO grade 2 meningiomas exhibit highly heterogeneous clinical courses. While the Ki67 proliferation index is a standard biomarker, its prognostic utility remains limited by methodological inconsistency and potential time-dependent dynamics. We evaluated an automated, artifact-adjusted Ki67 assessment and its integration with molecular risk profiling. 98 WHO grade 2 meningiomas (WHO 2021) were analyzed using an automated QuPath-based pipeline with HistoART for artifact exclusion. Molecular risk was defined by methylation and copy number profiling to calculate the integrated molecular-morphologic risk score by Maas et al. We employed extended Cox models to account for proportional hazards violations. Automated Ki67 values were significantly lower than routine pathological estimates (median 2.91% vs. 10%; p < 0.001) and correlated modestly with integrated risk scores (ρ = 0.26, p = 0.009). We identified a biphasic risk pattern: within the first 38 postoperative months, an automated Ki67 > 3.62% was a strong independent predictor for local recurrence (HR 5.06, p < 0.001) and progression-free survival (HR 4.15, p = 0.002), remaining significant alongside subtotal resection and the integrated risk group. Beyond 38 months, prognostic impact attenuated. Ki67 and the integrated molecular risk score contributed independently in multivariable models, suggesting complementary biological dimensions. Automated, artifact-adjusted Ki67 quantification provides time-dependent, independent prognostic information in WHO grade 2 meningioma, complementary to molecular risk stratification. It may serve as a cost-effective surveillance marker-both as an adjunct to molecular profiling and as a standalone tool where molecular testing is unavailable.

Complete hydatidiform mole with a coexisting live fetus in a twin pregnancy is exceptionally rare and carries major maternal and fetal risks. We describe two such pregnancies to illustrate management options. The first followed frozen-thawed single blastocyst transfer and presented at 13 weeks with a live fetus, a separate molar mass, an intramural fibroid, markedly elevated serum beta-human chorionic gonadotropin, and biochemical hyperthyroidism. Multidisciplinary assessment led to hysterotomy with removal of the fetus, molar tissue, and fibroids, followed by rapid recovery and no gestational trophoblastic neoplasia. The second was a spontaneously conceived twin pregnancy with a live 9-week fetus and multicystic molar tissue that was evacuated by suction curettage. An initial decline in beta-human chorionic gonadotropin was followed by a low-level plateau that resolved after a single low-dose methotrexate cycle. These cases highlight the need for individualized, multidisciplinary care and close biochemical follow-up.

HPV18-positive cervical large cell neuroendocrine carcinoma (LCNEC) is exceptionally rare. We report a 40-years-old woman with FIGO 2018 stage IVB LCNEC who progressed rapidly despite paclitaxel-carboplatin-bevacizumab plus pembrolizumab, followed by concurrent chemoradiotherapy and later cytotoxic regimens. Biopsy showed chromogranin A and synaptophysin positivity, diffuse p16, and Ki-67%~80%. Comprehensive profiling revealed very low tumor mutational burden (TMB, 1.21/Mb), microsatellite stability, no homologous recombination deficiency, and minimal PD-L1 (~1%), consistent with a poorly immunogenic tumor microenvironment (TME). This profile may explain the limited effect of immune checkpoint inhibition in this histology. The case suggests that HPV or p16 positivity alone did not predict clinical benefit from pembrolizumab-based therapy in advanced cervical LCNEC, and supports careful biological assessment before treatment selection. These observations provide practical guidance for clinicians and support evaluation of alternative or investigational strategies for this aggressive tumor.

Colorectal cancer (CRC) is a leading cause of cancer-related morbidity and mortality worldwide; thus, understanding its molecular mechanisms is critical for developing novel therapeutic targets. Long non-coding RNAs (lncRNAs) play crucial isoform-specific roles in cancer. While oncogenic lncRNA LINC00673 is known to be involved in multiple malignancies and possesses five distinct transcript variants, the functional role of LINC00673-V4-a highly expressed transcript variant in CRC-remains largely unexplored. This study investigated how LINC00673-V4 drives CRC proliferation by modulating the Hippo-Yes-associated protein (Hippo-YAP) signaling pathway.

Survival analysis was performed using the GSE39582 dataset to assess the prognostic value of LINC00673 in patients with CRC. The expression of LINC00673 transcript variants in CRC cell lines was detected by quantitative PCR. Cell Counting Kit-8 and 5-ethynyl-2'-deoxyuridine (EdU) incorporation assays were used to assess cell proliferation upon LINC00673-V4 knockdown or overexpression. Western blotting and immunofluorescence staining were utilized to analyze the expression of Hippo-YAP target genes. To confirm the interaction between LINC00673-V4 and fused in sarcoma (FUS), RNA immunoprecipitation (RIP) was performed, while co-IP was used to investigate potential protein-protein interactions.

High LINC00673 expression was observed in CRC cell lines and was associated with a poor prognosis in patients with CRC. Among the five LINC00673 transcript variants, LINC00673-V3 and LINC00673-V4 were the predominantly expressed isoforms; however, only LINC00673-V4 significantly promoted CRC cell proliferation. LINC00673-V4 inhibited YAP phosphorylation at Ser127, promoted YAP nuclear translocation, and upregulated the expression of Hippo-YAP target genes (connective tissue growth factor, cysteine-rich angiogenic inducer 61, survivin). RIP assays confirmed an association between LINC00673-V4 and FUS, while co-immunoprecipitation (co-IP) assays revealed that FUS interacted with both YAP and large tumor suppressor 1 (LATS1), suggesting FUS may facilitate LATS1-mediated YAP phosphorylation. FUS was found to promote LATS1-mediated YAP Ser127 phosphorylation, induce YAP cytoplasmic retention, and down-regulate Hippo-YAP target gene expression. Rescue experiments showed that LINC00673-V4 reversed the FUS-induced suppression of Hippo-YAP target gene expression.

Our study identified LINC00673-V4 as an isoform-specific oncogenic lncRNA in CRC. By associating with FUS, LINC00673-V4 sequesters FUS away from the LATS1/YAP complex; thereby inhibiting FUS-mediated YAP phosphorylation and enhancing Hippo-YAP target gene expression. These findings expand the landscape of lncRNA isoform-specific regulation in cancer, highlighting LINC00673-V4 as a potential prognostic biomarker and therapeutic target for CRC.

Gut microbiota dysbiosis is a crucial driver of the initiation and progression of colorectal cancer (CRC), where functional gut microbes and their metabolites play key roles in the microecological regulation of CRC. Currently, the association between Lachnospiraceae and CRC progression, as well as the underlying mechanisms, remains incompletely understood and warrants further investigation.

Bioinformatics analysis was performed to explore the co-pathway association between gut microbiota and metabolites in CRC patient samples. In vivo animal models were established to assess the regulatory effects of Lachnospiraceae on CRC tumorigenesis and gut microbiota homeostasis. The anti-CRC activities of Lachnospiraceae and its metabolite malate were investigated using in vitro experiments that measured cell viability, proliferation, apoptosis, and colony formation. Western blotting was performed to detect the expression levels of key proteins in the Wingless/Integrated (Wnt)/β-catenin signaling pathway.

Bioinformatics analysis revealed that malate was significantly downregulated in CRC patients, accompanied by gut microbiota dysbiosis driven predominantly by short-chain fatty acid (SCFA)-related Firmicutes such as Lachnospiraceae and Ruminococcaceae. In vivo, Lachnospiraceae restored gut microbiota homeostasis, reduced tumor number, and decreased tumor load. In vitro, Lachnospiraceae suppressed colorectal tumorigenesis and increased colonic D-malate levels.

Lachnospiraceae bacterium biologics abstracts accession-2278 (BAA-2278) is associated with anti-CRC effects in preclinical models, potentially mediated through regulation of gut microbiota homeostasis and inhibition of Wnt/β-catenin signaling via its metabolite malate.

Multiple myeloma (MM) is a heterogeneous hematological malignancy characterized by the clonal proliferation of plasma cells in the bone marrow, with distinct subtypes including smoldering MM (SMM), newly diagnosed MM (NDMM), and relapsed/refractory MM (RRMM). Despite therapeutic advances, outcomes remain unsatisfactory, especially for RRMM, due to unclear heterogeneity, progression mechanisms, and crosstalk between tumor cells and the bone marrow microenvironment (BMME) via direct interactions or extracellular vesicles (EVs).

Single-cell RNA sequencing (scRNA-seq) was performed on bone marrow samples from 12 MM patients, and major cell types were identified. Plasma cells were subjected to re-clustering to explore subpopulation heterogeneity. Functional enrichment analysis of differentially expressed genes (DEGs) was conducted. Cellular stemness was evaluated by CytoTRACE, and developmental trajectories were inferred via Monocle and Slingshot. Cell-cell communication was analyzed by CellChat, while transcription factor (TF) regulatory networks were identified through SCENIC analysis. Metabolic pathway activity was also assessed. Finally, loss-of-function experiments (siRNA-mediated ASS1 knockdown) were conducted to validate its functional role.

The analysis identified 7 major cell types. Plasma cells were further stratified into 6 subpopulations (C0-PCSK1N+, C1-IGHGP+, C2-IGHA1+, C3-ASS1+, C4-CD27+, C5-STMN1+). C3 and C5 were enriched in RRMM, while C1 and C4 were dominant in SMM. C3 exhibited hyperactive EV signature and higher stemness-like scores. Pseudotime trajectory analysis identified C3 as poorly differentiated malignant progenitors driving disease progression. C3 showed strong crosstalk with monocytes/macrophages/conventional dendritic cells (cDCs) via MIF and ICAM signaling. Key TFs regulating C3 included ATF5, TP73, MYB, CEBPB, and NFIA. Metabolic pathway analysis indicated enhanced vitamin B6 metabolism, phenylalanine metabolism, and oxidative phosphorylation in C3 and RRMM. ASS1 silencing inhibited proliferation, clonogenic capacity and migration, while promoting apoptosis in MM cells.

Our study delineates MM heterogeneity, developmental dynamics, and regulatory networks at the single-cell level. ASS1+ plasma cells represent a highly malignant subpopulation associated with RRMM, driving disease progression through unique TF regulatory networks, metabolic reprogramming, and crosstalk with the BMME. These findings provide novel insights into MM pathogenesis and identify ASS1 as a potential therapeutic target for MM patients, particularly those with RRMM.

Regulatory T cells (Tregs) are central to immune regulation, preventing excessive immune responses, maintaining immune tolerance, and modulating inflammatory microenvironments. Dysregulation of Treg development, differentiation, proliferation, or function contributes significantly to autoimmune diseases, inflammatory disorders, and tumor immune evasion. Protein kinases, key mediators of cellular signaling, regulate diverse processes including motility, metabolism, transport, and cell cycle progression; their emerging roles in immune regulation make them promising therapeutic targets for inflammatory diseases, autoimmunity, and immunotherapy-treated cancers. Notably, protein kinases modulate Treg differentiation and function by controlling the expression of the lineage markers Forkhead box protein 3 (Foxp3, intracellular) and CD25 (cell surface). This mechanistic review addresses: (1) fundamental Treg characteristics, functions, and disease relevance; (2) protein kinase-mediated regulatory mechanisms in Tregs; and (3) progress on protein kinase inhibitors for Treg-related diseases. The review aims to outline the kinase regulatory network governing Treg biology and guide future identification of kinase targets for treating autoimmune, inflammatory, and malignant diseases.

Galectins, a structurally conserved family of β-galactoside-binding lectins, have emerged as key regulators of cancer progression. However, their integrated roles across tumor immunity, metabolic rewiring, and metastasis have yet to be fully defined. In this review, we synthesize current evidence on major galectin family members, particularly galectin (Gal)-1, Gal-3, Gal-4, Gal-7, Gal-9, and Gal-13, and delineate how they drive tumor progression through mechanistically distinct yet convergent pathways. Specifically, galectins induce T-cell dysfunction through T-cell immunoglobulin and mucin-domain containing-3 (TIM-3)- and programmed cell death protein 1 (PD-1)-associated signaling, reprogram macrophages and myeloid-derived suppressor cells (MDSCs) via phosphoinositide 3-kinase/protein kinase B (PI3K/AKT), Janus kinase/signal transducer and activator of transcription (JAK/STAT), and nuclear factor kappa B (NF-κB) pathways, and modulate innate immune effectors, including natural killer (NK) cells, neutrophils, and dendritic cells, in a context-dependent manner. Beyond immune regulation, galectins reshape tumor metabolism through effects on glycolysis, lipid metabolism, and glycan remodeling, and differentially regulate ferroptosis susceptibility, with the contrasting roles of Gal-1 and Gal-13 underscoring functional diversity within the family. Emerging evidence further implicates galectins in resistance to chemotherapy, targeted therapy, immune checkpoint blockade, and chimeric antigen receptor T-cell (CAR-T) therapy, positioning them as candidate therapeutic targets. We also discuss galectin-directed strategies, including small-molecule inhibitors, nanomedicine-based platforms, vaccination approaches, and rational combination therapies, and highlight the promise of multi-galectin biomarker panels for precision oncology.

Glioblastoma (GBM) is a highly malignant tumor with poor prognosis and limited effective treatment options. Emerging studies have suggested that gut microbiota may influence glioma progression through the gut-brain axis, though the precise mechanisms remain largely unclear. In this study, we employed a comprehensive multi-omics approach-encompassing single-cell transcriptomics, spatial transcriptomics, metagenomics, metabolomics, and m6A-seq-to investigate how antibiotic-induced gut microbiota disruption impacts glioma progression in a mouse model. Gene expression analysis revealed significant alterations in antibiotics-treated mice (ABX-treated mice), including reduced expression of Epha6 and upregulated expression of Tead1, key genes associated with glioma progression and immune modulation. Spatial transcriptomics and metabolomic profiling identified reduced methionine levels in gliomas of ABX-treated mice, linking gut-derived metabolite changes to epigenetic regulation via m6A methylation. Single-cell RNA sequencing further demonstrated an increased proportion of AC-like cells, disrupted intercellular communication, and aberrations in the EPHA and NRXN signaling pathways. These findings highlight the interplay between gut microbiota, immune signaling, and epigenetic modifications in shaping the glioma microenvironment. This study advances our understanding of the gut-brain axis in glioma biology and proposes the EPHA pathway as a promising biomarker for the immune-mediated modulation of tumor progression, thereby providing new insights into the role of the gut-brain axis in glioma regulation.