A miR-99a-TGF-β1 feedback circuit drives miR-99a upregulation.

Background This study investigated the role of peptidylprolyl isomerase A (PPIA) in lung adenocarcinoma (LUAD) using bioinformatics approaches and experimental validation. Methods This study began by assessing PPIA expression and its prognostic significance in LUAD. We then performed functional enrichment analyses and examined CNVs, DNA methylation, DNMT activity, RNA modification, immune cell infiltration, and drug therapy efficacy. Cellular experiments were performed to confirm the role of PPIA in DNA damage repair. Results PPIA was overexpressed in LUAD tissues and was associated with poor prognosis, including shorter overall survival and progression-free interval. PPIA expression was correlated with CNVs, DNA methylation, DNMT activity, and RNA modification-related genes. It was also negatively associated with immune cell infiltration and immune-related gene expression, suggesting a potential immunosuppressive role. Drug sensitivity analysis indicated that PPIA may serve as a predictive biomarker for therapeutic efficacy in LUAD. In addition, PPIA expression was associated with DNA damage repair-related genes. Cellular experiments show that PPIA knockdown increases radiosensitivity and γ-H2AX foci formation, whereas PPIA overexpression exerts the opposite effects. Conclusion This study highlights the potential of PPIA as a prognostic biomarker and target in LUAD, particularly in the regulation of DNA damage repair and tumor immune microenvironment remodeling.

RNA processing and modification are critical for virus replication and pathogenesis, yet how the host exploits viral RNA features, particularly the poly(A) tail-for antiviral defense remains unclear. Through multi-omics integration and systematic functional screening, we identify the poly(A)-binding protein PABPC1 as a broad-spectrum restriction factor against multiple coronaviruses. We demonstrate that PABPC1 preferentially binds viral RNAs bearing short poly(A) tails-distinct from the longer and more heterogeneous host poly(A) tails and, in a poly(A)-length-dependent manner, recruits the mitochondrial exonuclease EXD2 to assemble a degradative RNA-protein complex. Both the recognition and subsequent degradation of viral RNA strictly depend on poly(A) tail length, enabling selective decay of short-tailed viral transcripts while sparing host mRNAs. Importantly, inflammatory signaling enhances the expression of PABPC1 and EXD2, suggesting its role as an inducible host defense pathway activated during viral infection. Capitalizing on this discovery, we engineered a synthetic fusion protein mimicking the PABPC1-EXD2 complex and achieved efficient delivery using lipid nanoparticles (LNPs). This rationally designed therapeutic exhibits robust suppression of coronavirus replication in both cellular and murine models. Collectively, our findings uncover a novel host antiviral strategy that targets a conserved viral RNA structural element and provide a conceptual framework for developing host-derived and broad-spectrum anti-coronavirus therapeutics. ### Competing Interest Statement The authors have declared no competing interest. National Key R&D Program of China, 2021YFA1300800 China NSFC projects, 82341061, 82502690 Fundamental Research Funds for the Central Universities, 2042022dx0003

Background/Objectives: Poly(lactic-co-glycolic acid) (PLGA) microspheres offer sustained drug delivery but often suffer from broad particle size distribution (PSD), leading to inconsistent release profiles. This study investigates wet sieving as a post-processing strategy to precisely control PSD, quantified by the Span value, and evaluates its impact on the performance of triamcinolone acetonide (TA)-loaded PLGA microspheres. Methods: Triamcinolone acetonide-loaded PLGA microspheres were prepared via emulsification-solvent evaporation. Wet sieving was employed as a post-processing strategy to obtain distinct particle size fractions and groups with defined polydispersity (Span values). The microspheres were characterized for particle size distribution, drug loading, surface morphology, and in vitro release kinetics. To establish the in vivo relevance of polydispersity control, the pharmacokinetic profiles of different Span groups were first determined using LC-MS/MS following intra-articular injection in rats. Subsequently, their therapeutic efficacy was evaluated in a rat model of knee osteoarthritis, with outcomes assessed by joint swelling measurement and histopathological analysis. Results: Microspheres were prepared, fractionated into distinct size groups (0–20, 20–28, 28–40, 40–50, >50 μm) and polydispersity groups (Span = 1.4, 0.8, 0.5). We identified Span as a dominant factor independent of mean particle size. Reducing the Span from 1.4 to 0.5 significantly decreased burst release (24.15% to 14.51%), prolonged mean residence time (MRT 88.52 to 123.53 h), and enhanced anti-inflammatory and cartilage-protective effects in a rat model of knee osteoarthritis. Conclusions: This work establishes Span ≤ 0.5 as a critical quality attribute and presents wet sieving as a simple, effective method to ensure batch-to-batch consistency and predictable in vivo performance for PLGA microsphere products.

IntroductionThe emergence of new SARS-CoV-2 variants with immune evasion capabilities underscores the importance of developing a broad-spectrum and effective vaccine. The receptor binding domain (RBD) of the Spike protein has been widely utilized in vaccine due to its high immunogenicity. However, the Spike protein, particularly the RBD region, exhibits significant variability in the evolution of SARS-CoV-2, leading to viral immune evasion and reduced vaccine effectiveness.MethodsA broad-spectrum antigen (M5-RBD) was developed via mutation patching, incorporating key high-impact mutation sites (K417T, L452R, T478K, E484K, N501Y). Additionally, extra mutations (N440K or G446S) were introduced into M5-RBD to evaluate their impact on immune response. M5-RBD was further combined with a novel CpG adjuvant HP007 for immunization.ResultsM5-RBD elicited high titers of broad-spectrum neutralizing antibodies against SARS-CoV-2 wild-type and various variants (Delta, Omicron BA.1, BA.2, BA.2.75, BA.5, BF.7, BQ.1.1, XBB, EG.5, JN.1, KP.3 strains). Introduction of N440K or G446S significantly diminished the immune response to viral strains. When combined with HP007 adjuvant, M5-RBD induced efficient and durable T cell responses, providing protection to K18-hACE2 KI mice against lethal infections with both wild-type and Omicron BA.2 strains.DiscussionRationally designed with key high-impact mutation sites, M5-RBD effectively overcomes SARS-CoV-2 variant immune evasion and elicits broad-spectrum neutralizing antibodies. The combination with HP007 adjuvant enhances immune protection, providing a promising strategy for the development of next-generation COVID-19 vaccines.

EV-miR-99a targets NLRP2 and activates NF-kB signaling in fibroblasts.

PURPOSE:Antibody-drug conjugate (ADC) targeting human epidermal growth factor receptor 2 (HER2) could be a promising strategy for HER2-expressing gastric cancer or gastroesophageal junction adenocarcinoma (GC/GEJ) and colorectal cancer (CRC). We conducted a phase I trial to assess trastuzumab rezetecan, a novel HER2-targeted ADC, in HER2-expressing advanced GC/GEJ and CRC. METHODS:Patients with HER2-expressing advanced GC/GEJ and CRC whose disease progressed on and/or had no available/applicable standard treatment were enrolled. Patients were intravenously given trastuzumab rezetecan at 3.2, 4.8, 6.4, and 8.0 mg/kg (once every 3 weeks) in an i3+3 dose-escalation scheme, followed by pharmacokinetics expansion at selected doses and then clinical expansion. The primary end points were dose-limiting toxicity (DLT) and safety. RESULTS:Between March 30, 2021, and August 1, 2023, 100 patients were enrolled (57 with GC/GEJ and 43 with CRC). One DLT occurred in the 8.0 mg/kg dose cohort. Grade ≥3 treatment-related adverse events (TRAEs) were reported in 66 (66.0%) patients. Only 5 (5.0%) patients discontinued treatment because of TRAEs. In HER2-positive GC/GEJ (n = 40), trastuzumab rezetecan achieved an objective response rate (ORR) of 45.0%, a median progression-free survival (PFS) of 9.0 months (95% CI, 7.0 to 11.3), and a median overall survival (OS) of 16.3 months (95% CI, 12.4 to not reached [NR]). In GC/GEJ with HER2 immunohistochemical 2+ and in situ hybridization-negative (n = 12), trastuzumab rezetecan had an ORR of 25.0%, a median PFS of 12.2 months (95% CI, 2.8 to 14.0), and an immature median OS. In HER2-positive CRC (n = 37), trastuzumab rezetecan had an ORR of 40.5%, a median PFS of 9.5 months (95% CI, 7.3 to 11.2), and a median OS of 22.7 months (95% CI, 17.5 to NR). CONCLUSION:Trastuzumab rezetecan showed tolerable safety and preliminary efficacy in HER2-expressing advanced GC/GEJ and CRC.

Pancreatic adenocarcinoma (PDAC) is a highly lethal malignancy characterized by profound resistance to immunotherapy. Converting immunologically “cold” tumors into “hot” tumors by enhancing T cell infiltration represents a promising therapeutic strategy, yet the vascular mechanisms regulating immune recruitment in PDAC remain poorly defined. Here, we integrated single-cell RNA sequencing with GEPIA analysis and spatial transcriptomics to investigate the functional role of Ras Interacting Protein 1 (RASIP1)-positive endothelial cells in PDAC. We identified RASIP1-positive tumor endothelial cells as a distinct endothelial subpopulation enriched in leukocyte transendothelial migration pathways, with upregulated adhesion molecules and spatial co-localization with T-effector and IFN-γ signatures. Multiplex immunohistochemistry of human PDAC tissues revealed prominent perivascular accumulation of CD8⁺/GranzymeB⁺ cytotoxic T lymphocytes surrounding RASIP1-positive vessels. Mechanistically, RASIP1 knockdown reduced ICAM1 expression, whereas RASIP1 overexpression enhanced ICAM1 signaling, and both modulated ERK phosphorylation dynamics, suggesting that RASIP1 regulates endothelial functionality through ERK-related signaling. Collectively, our findings identify a distinct endothelial state that actively shapes the immune microenvironment of PDAC. Targeting RASIP1-positive endothelial cells may represent a potential strategy to enhance tumor immunogenicity and improve responsiveness to immunotherapy.

The tumor microenvironment is crucial for cancer progression, but the mechanisms underlying the tumor-immune cell interactions in it remain poorly understood. Here, we identified latent transforming growth factor-β (TGFβ) binding protein 4 (LTBP4) deficiency in colorectal cancer (CRC) as a critical driver that reprogrammed tumor-associated macrophages (TAMs) and induced a distinct subset, which promoted tumor progression by coordinating immune evasion and extracellular matrix (ECM) remodeling. Clinically, LTBP4 deficiency correlated with CRC progression and poor patient survival. Ltbp4 knockout markedly promoted tumor growth and metastasis in immunocompetent mice, an effect attenuated in immunodeficient hosts, establishing the essential role of host immunity in mediating the effects of LTBP4 deficiency. Single-cell RNA sequencing revealed that LTBP4 deficiency induced a mannose receptor C-type 1-positive (MRC1+)/CD44+ TAM subset and correlated with reduced CD8+ T cell infiltration. Mechanistically, LTBP4 deficiency increased active TGFβ1 levels, which acted in a paracrine manner to up-regulate MRC1 in TAMs, whereas autocrine signaling induced HAS2 (hyaluronan synthase 2) expression and hyaluronan production to increase CD44. CD44 signaling in TAMs up-regulated matrix metalloproteinases for collagen degradation, whereas MRC1 mediated collagen internalization, cooperatively remodeling the ECM to facilitate tumor invasion. The TGFβ1-driven MRC1+/CD44+ TAMs further suppressed CD8+ T cell function by diminishing the C-X-C motif chemokine ligand 16-C-X-C motif chemokine receptor 6 (CXCL16-CXCR6) axis. Therapeutically, targeted depleting MRC1+/CD44+ TAMs enhanced the efficacy of PD-1 (programmed cell death-1) blockade in LTBP4-deficient tumors. Our study positions LTBP4 as a key modulator of tumor progression and reveals a therapeutic strategy for LTBP4-deficient CRC.

ABSTRACT:Arginine methylation catalyzed by protein arginine methyltransferases (PRMTs) is required for cancer cell proliferation, but whether PRMTs mediate resistance to therapy remains unclear. Here, we performed loss-of-function screens in venetoclax-resistant (VEN-R) acute myeloid leukemia (AML) patient-derived xenograft cells and found that PRMT9 plays a critical role in promoting VEN resistance. Specifically, VEN-R AML samples exhibited high levels of PRMT9, and PRMT9 inhibition resensitized AML cells to VEN treatment. In preclinical resistant models, genetic ablation of PRMT9 synergized with VEN to eradicate AML cells. Consistently, pharmacologic inhibition of PRMT9 combined with VEN produced similar effects in VEN-R AML mouse models. Mechanistically, PRMT9 ablation disrupted RNA splicing by inducing exon skipping in mRNA encoding ALG13, an uridine diphosphate (UDP)-N-acetylglucosaminyltransferase subunit, thereby downregulating expression of the VEN efflux transporter encoded by the adenosine triphosphate-binding cassette subfamily C member 1 gene. PRMT9 inhibition also suppressed protein synthesis, leading to downregulation of short-lived oncoproteins such as MCL1. These findings establish a connection between PRMT9-mediated arginine methylation and poor VEN responsiveness and demonstrate that targeting PRMT9 may represent a viable strategy to overcome VEN resistance.

As an emerging branch of clinical medicine, microbiota medicine has attracted worldwide attention from clinicians, medical educators, patient communities, and industry. However, this developing field still lacks consensus on its fundamental principles as well as guidelines for clinical and educational practice. An expert panel was convened by the journal Microbiota Medicine Research at the 2025 CHINAGUT Conference to develop the principles and practice guidelines of microbiota medicine using the Delphi method. This document provides a new framework for clinicians, educational institutions, and healthcare administrators. The expert panel developed 15 key statements, encompassing definitions of microbiota medicine and dysbiosis-related diseases, graded value evaluation of microbiome testing technologies, pathways for multidisciplinary discussions on complex dysbiosis-related diseases, and educational frameworks for physicians in microbiota medicine. The panel further recommends incorporating microbiota medicine into undergraduate and postgraduate medical education and emphasizes the application of artificial intelligence in supporting microbiota medicine. This guideline defines core competencies required for physicians specializing in this discipline. Collectively, this guideline aims to define the significant role of microbiota medicine in clinical practice and medical education, thereby advancing its sustainable development.

miR-99a associates with CRC progression and prognosis.

Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by persistent synovial inflammation and progressive bone destruction in which immune dysregulation plays a central role. Recent evidence has highlighted the gut–bone axis as a critical framework linking gut microbiota to skeletal and immune homeostasis. Gut microbiota dysbiosis disrupts intestinal barrier integrity by altering tight junction proteins and increasing intestinal permeability, facilitating microbial translocation and triggering systemic inflammatory responses. Microbiota-derived metabolites, including short-chain fatty acids, bile acids, and tryptophan metabolites, act as key mediators along the gut–bone axis. These metabolites regulate multiple signaling pathways and immune cell functions, particularly by modulating the balance between T helper 17 and regulatory T cells, suppressing B-cell hyperactivation, promoting macrophage M2 polarization, and inhibiting dendritic cell maturation. These actions may contribute to immune homeostasis and bone metabolism associated with RA. This review systematically summarizes the role of gut microbiota dysbiosis, intestinal barrier dysfunction, and microbial metabolites in RA pathogenesis within the framework of the gut–bone axis. Furthermore, microbiota-targeted therapeutic strategies, including probiotics, prebiotics, dietary interventions, fecal microbiota transplantation, and traditional Chinese medicine, are discussed as potential approaches to restore host-microbiota balance. However, most current evidence is derived from preclinical studies, highlighting the need for further clinical validation. Despite these limitations, a deeper understanding of microbiota-driven mechanisms along the gut–bone axis may provide novel insights into RA pathogenesis and facilitate the development of targeted and personalized therapeutic strategies.

EVs from highly metastatic CRC cells promote fibroblast activation.

This study aims to compare the efficacy of modified transumbilical single-port and conventional 3-port laparoscopic minimally invasive cholecystectomy based on the levels of inflammatory factors and serum amylase. Ninety-one patients with benign gallbladder diseases admitted to our hospital from January 2023 to December 2024 were retrospectively analyzed. Patients were divided into group A (n = 45) and group B (n = 46) according to the surgical method. The group A accepted traditional laparoscopic cholecystectomy and the group B accepted modified transumbilical single-port laparoscopic cholecystectomy. The perioperative indicators, degree of pain, levels of inflammatory factors, liver function, serum amylase level, immune function, patients' satisfaction with aesthetic incision and incidence of complications were compared in both groups. Compared with group A, group B was associated with a longer operative time (95% confidence interval: -15.9 to -1.5; P = .018). Group B demonstrated significantly lower pain scores at both 12 hours (postoperative day [POD] 0.5) and 24 hours (POD 1) postoperatively (both P <.001), with multivariate analysis confirming these differences remained significant after adjustment for covariates (P <.001). While both groups showed postoperative elevations in inflammatory markers [high-sensitivity C-reactive protein (hs-CRP), interleukin-8 (IL-8), IL-2, tumor necrosis factor-alpha (TNF-α)], liver enzymes [total bilirubin (TBIL), aspartate aminotransferase (AST), glutamyltranspeptidase (GGT)], and serum amylase, Group B maintained significantly lower hs-CRP levels after adjustment (P = .018). Immunological assessment revealed better-preserved immunoglobulin levels [Immunoglobulin A (IgA), IgG, IgM] in Group B at POD 7 (all P <.001). Patients receiving the single-port technique reported higher cosmetic satisfaction (95% confidence interval: -0.88 to -0.45, P <.001) and experienced fewer complications (P <.05). No significant intergroup differences were observed in recovery milestones (first flatus, first defecation), hospital stay, or intraoperative blood loss (all P >.05). The modified transumbilical single-port laparoscopic cholecystectomy may reduce the degree of pain, reduce the inflammatory response, reduce serum amylase level, improve the liver function and immune function, promote patients' satisfaction with aesthetic incision and reduce the incidence of complications. However, the retrospective design limits causal inferences and requires validation by prospective studies.

Immune checkpoint inhibitors (ICIs) are the preferred treatment for advanced hepatocellular carcinoma (HCC). However, most patients do not respond to initial immunotherapy alone, even when combined with other therapies. Our study aimed to profile ICI-based therapies for HCC patients in real-world clinical practice and to identify factors associated with survival and treatment efficacy. A retrospective cohort study was conducted to describe ICI-based therapies for HCC patients at our center, including combination modalities, effectiveness, and safety. Pre- and post-treatment indicators were collected to explore factors related to progression-free survival (PFS) and treatment efficacy in these patients. All 110 patients received therapy based on immune checkpoint inhibitors (ICIs) using three strategies, with the majority (60.9

The association between folate metabolism abnormalities and the development of colorectal cancer (CRC) remains controversial. Here, we report that the folate exerts a tumor-suppressive role in CRC; however, the manifestation of this effect is restricted by the expression level of folate transporter SLC46A1 in CRC cells. Multi-cohort profiling revealed significant downregulation of SLC46A1 in CRC tissues compared to adjacent normal tissues, where low expression independently predicted poor overall survival. Functional studies demonstrated that SLC46A1-mediated folate uptake suppressed tumor proliferation, migration, and invasion both in vitro and in vivo. Mechanistically, SLC46A1 deficiency restricted intracellular folate availability and impaired cellular methylation potential, as evidenced by a reduced SAM/SAH ratio, leading to DNA hypomethylation at specific sites such as the FOS proto-oncogene promoter. This epigenetic reprogramming triggers transcriptional activation of key oncogenic effectors CCND1, BCL2, and PLAU involved in CRC progression. Clinically, we found a significant inverse correlation between SLC46A1 expression and folate levels in tumor interstitial fluids of CRC, suggesting impaired folate uptake in low SLC46A1 tumors. Multi-color immunofluorescence across two cohorts further demonstrated conserved inverse associations between SLC46A1 and FOS expression in primary tumors and metastatic lesions. This study elucidates the molecular mechanism by which folate inhibits CRC progression through the “SLC46A1-epigenetic-transcriptional regulation” axis, providing mechanistic insights into folate deficiency-driven CRC progression and biomarkers for precision CRC intervention.

Safety assessment of LY2584702 in mice.

Extracellular matrix (ECM) remodeling contributes to retinal vascular basement membrane thickening, an early structural hallmark of diabetic retinopathy (DR). This study aimed to identify key ECM-related genes (ECMGs) associated with DR. Transcriptomic data of DR and ECMGs from MatrixDB were integrated to identify differentially expressed ECMGs. Six machine learning (ML) models, including Extra Trees (ET), Logistic Regression, Adaptive Boosting, Random Forest, Extreme Gradient Boosting, and naive Bayes classifier, were employed to construct DR classification models, with SHapley Additive exPlanation (SHAP) used to interpret feature contributions. Functional enrichment analysis using GSEA and immune infiltration analysis using CIBERSORT were conducted to explore the potential mechanisms by which key ECMGs regulate DR. Regulatory networks were constructed using predicted miRNAs, lncRNAs, and transcription factors (TFs) via the ENCORI, miRWalk, and miRNet databases. Drug-key ECMGs-DM-related diseases interactions were further explored using the DGIdb and CTD databases. Nine candidate ECMGs were identified by overlapping 356 DM-associated DEGs, 1,626 DR-associated DEGs, and 1,023 ECMGs, including CILP2, FN1, DEFA3, COL17A1, CRISP3, TPSAB1, SFRP1, GPHA2, and ECM2. Among the six ML algorithms, the ET classifier exhibited the best overall performance, and five ECMGs (SFRP1, CILP2, FN1, TPSAB1, and ECM2) with non-zero SHAP values were retained as key genes. These genes showed distinct expression patterns across the healthy, DM, and DR groups, and were enriched in neural-related pathways, such as axon guidance, glycosphingolipid biosynthesis ganglio series, and neuroactive ligand receptor interaction. Immune profiling and correlation analysis revealed that FN1, TPSAB1, and CILP2 were correlated with memory/naive B cells, CD8 + T cells, activated memory CD4 + T cells, Tregs, monocytes, and neutrophils. Additionally, the ceRNA network contained five miRNAs, 7 lncRNAs, and two ECMGs, and further regulatory and pharmacologic analysis further linked key ECMGs to specific TFs, drugs, and diabetes-related diseases. This study identified SFRP1, CILP2, FN1, TPSAB1, and ECM2 as key ECMGs in DR, revealing their coordinated involvement in ECM remodeling, neural signaling, and immune modulation. These findings provide novel insights into DR pathogenesis and potential therapeutic targets.