The repair of alveolar bone is of vital importance for maintaining oral health and promoting recovery following injury or disease. Functioning as an NAD+-dependent protein deacetylase, SIRT1 modulates diverse physiological activities, with particular relevance to metabolism and osteogenesis. However, its specific role in alveolar bone repair and the associated metabolic pathways have not been fully elucidated. In this study, we explored the function of SIRT1 in alveolar bone healing using a conditional knockout mouse model (Wnt1-Cre; SIRT1fl/fl mice) and evaluated its involvement in glycolytic metabolism through the Wnt/β-catenin signaling pathway. The deletion of SIRT1 resulted in significantly impaired bone healing within extraction sockets. Notably, bioinformatics analysis suggested that SIRT1 deficiency may alter the metabolic profile of orofacial mesenchymal stem cells (OMSCs). Consistently, glycolytic activity was markedly reduced in SIRT1-deficient OMSCs, as evidenced by decreased extracellular acidification rate (ECAR), reduced lactate production, and lower expression levels of glycolytic enzymes. Mechanistically, we demonstrated that SIRT1 interacts with β-catenin and that SIRT1 deficiency is associated with increased β-catenin acetylation and reduced nuclear localization, thereby impairing Wnt/β-catenin signaling and glycolytic metabolism. Both in vivo and in vitro rescue experiments using SKL2001, a Wnt/β-catenin signaling pathway agonist, revealed that SKL2001 was able to restore β-catenin nuclear translocation, enhance glycolytic metabolism, and improve the impaired osteogenic differentiation caused by SIRT1 deficiency. The results of this study highlight a previously unidentified role of SIRT1 in promoting alveolar bone repair by modulating glycolysis through the Wnt/β-catenin pathway. These findings not only advance our understanding of bone repair at the metabolic level but also propose SIRT1 and Wnt signaling as viable therapeutic avenues.

BACKGROUND:Myelodysplastic neoplasms (MDS) represent a group of heterogeneous clonal disorders characterized by immune dysregulation in their pathogenesis. Gut microbiota dysbiosis plays a critical role in immune modulation. METHODS:We collected the fecal samples of 23 newly diagnosed MDS, 10 hypomethylating agents (HMA) treated MDS and 13 age and sex matched healthy controls (HC), and analyzed the gut microbiota compositions and functional pathways using metagenomic next-generation sequencing (mNGS). RESULTS:Distinct microbial compositions were observed between newly diagnosed MDS and HC. Notably, the Veillonellaceae family was significantly enriched in MDS patients. Specific bacteroid species demonstrated significant correlations with lymphocyte subtypes, functional activation status, and serum inflammatory cytokines. Functional profiling revealed altered metabolic pathways in newly diagnosed patients, particularly in amino acid metabolism and ATP synthesis. Notably, glutamine/glutamate and tryptophan metabolism pathways were hyperactive in untreated MDS but downregulated following HMA treatment. CONCLUSIONS:The gut microbiota altered in MDS patients and was associated with immune dysregulation and inflammation, which may contribute to MDS pathogenesis and mediate therapeutic effects of HMA treatment, highlighting the gut microbiota-metabolism axis as a potential therapeutic target for MDS management.

Recently, mitophagy-mediated bone mineralization of mesenchymal stem cells has emerged as another bone formation pattern, but whether mitophagy-mediated bone mineralization shapes craniofacial development remains unknown. Here, we demonstrate that loss of OPTN, a keystone macroautophagy/autophagy receptor, impairs mitophagy and acidic calcium phosphate (ACP) transport in orofacial bone mesenchymal stem cells (OMSCs), leading to craniofacial bone mineralization defects. We substantiate that OPTN undergoes LLPS both in vitro and in vivo, driven by S173 phosphorylation within its intrinsically disordered N-terminal domain (NTD), facilitating the association of OPTN complexes with phagophore membranes. Additionally, the ubiquitin-binding domain (UBD) in OPTN's C-terminal domain (CTD) also promotes LLPS to recruit ubiquitin-modified mitochondria. Physiochemically, mutations at the conserved sites in human OPTN (S173A and D474N) disrupt the OPTN LLPS, as validated in mouse and zebrafish, thereby inhibiting mitophagy and impairing bone mineralization. Together, our findings reveal a new mechanism through which OPTN LLPS couples mitophagy-mediated mineralization to craniofacial bone development, highlighting its potential as a therapeutic target for treating orofacial malformations via modulation of mitophagy.Abbreviations: 1, 6HD: 1, 6-hexanediol; ACP: acidic calcium phosphate; ALP: alkaline phosphatase; ARS: Alizarin Red staining; BFR/BS: bone formation rate per bone surface; Baf-A1: bafilomycin A1; CCCP: carbonyl cyanide 3-chlorophenylhydrazone; CTD: C-terminal domain; dpf: days post-fertilization; EDS: energy dispersive spectroscopy; FL: full length; FRAP: fluorescence recovery after photobleaching; hpf: 24h post-fertilization; IDR: intrinsically disordered region; IHC: immunohistochemistry; LLPS: liquid-liquid phase separation; LC-MS/MS: liquid chromatography-tandem mass spectrometry; MAR: mineral apposition rate; MS/BS: mineralizing surface per bone surface; NTD: N-terminal domain; ODM: osteogenic differentiation medium; OMSCs: orofacial bone mesenchymal stem cells; OPTN: optineurin; P1: postnatal day 1; P21: postnatal day 21; PDB: Paget disease of bone; PTMs: post-translational modifications; qRT-PCR: quantitative real-time PCR; S173: serine 173; STK4: serine/threonine kinase 4; SEM: scanning electron microscopy; TMD: tissue mineral density; TEM: transmission electron microscopy; UBD: ubiquitin-binding domain; Ub: ubiquitin.

Evodiamine (EVO), a major bioactive alkaloid isolated from Tetradium ruticarpum (A.Juss.) T.G.Hartley, possesses diverse pharmacological activities; however, its potential hepatotoxicity remains insufficiently characterized. This study aimed to evaluate the hepatotoxic effects of EVO and explore the molecular events associated with its toxicity. Male mice were administered EVO (10, 20, or 40 mg/kg) for 7, 14, or 28 days. EVO exposure caused dose- and time-dependent liver injury and oxidative stress, as shown by serum biochemistry, histopathology, and oxidative stress markers. An integrated multi-omics strategy combining network toxicology, transcriptomics, and metabolomics was applied to explore the underlying mechanisms. EVO exposure induced significant liver injury and oxidative stress in a dose- and time-dependent manner. Multi-omics analyses suggested that EVO treatment was associated with alterations in pathways related to inflammation, apoptosis, and lipid metabolism, including FOXO, PPAR, and NF-κB signaling pathways. Changes in the expression of SIRT1, CASP2, and FOXO3 and disturbances in glycerophospholipid metabolism were further observed. In conclusion, EVO induces dose- and time-dependent hepatotoxicity in mice. Multi-omics analyses suggest that inflammatory responses, apoptotic processes, and metabolic disturbances may contribute to EVO-induced liver injury. These findings provide toxicological evidence regarding the safety profile of EVO and identify biological pathways associated with its hepatotoxicity.

Background Metabolic reprogramming within the tumor microenvironment is a pivotal barrier to effective immune checkpoint blockade (ICB). While programmed death ligand 1 (PD-L1) is well characterized as a ligand inhibiting T-cell function, its intrinsic ‘reverse signaling’ role in regulating tumor metabolism and shaping the immune landscape remains poorly understood. Here, we investigated the metabolic determinants of resistance to anti-programmed cell death protein 1 (anti-PD-1) therapy and the underlying molecular mechanisms.Methods Integrated metabolomics and transcriptomics were performed on tumor samples from patients with non-small cell lung cancer and cell lines. Mechanisms were delineated using RNA sequencing, cleavage under targets and tagmentation assays, metabolic flux analysis, and coculture systems. The therapeutic efficacy of targeting metabolic effectors was evaluated in syngeneic mouse models and correlated with immune profiling.Results We identified a distinct metabolic signature characterized by aberrant pyruvate accumulation in patients resistant to anti-PD-1 therapy. Mechanistically, we demonstrate that antibody-mediated ligation of PD-L1 triggers an intrinsic endoplasmic reticulum (ER) stress response via the PERK–ATF4–CHOP axis. ATF4 acts as a transcriptional activator that directly upregulates pyruvate dehydrogenase kinase 4 (PDK4) (blocking pyruvate oxidation) and glutaminase (GLS) (promoting glutaminolysis), creating a ‘dual-hit’ metabolic rewiring that drives intracellular pyruvate build-up. Subsequently, tumor-secreted pyruvate is taken up by tumor-associated macrophages (TAMs) via MCT1, inducing mitochondrial reactive oxygen species accumulation and driving them into a state of cellular senescence. These senescent TAMs upregulate PD-L1 via STAT3 signaling, thereby reinforcing an immunosuppressive feedback loop. Pharmacological inhibition of PDK4 and GLS effectively abolished pyruvate accumulation, prevented macrophage senescence, and restored CD8+ T-cell cytotoxicity.Conclusions Our study identifies a novel ‘PD-L1–ER stress–pyruvate–macrophage senescence’ axis as a key mechanism underlying primary resistance to ICB. These findings highlight the non-canonical reverse-signaling function of PD-L1 in metabolic remodeling and propose that targeting the PDK4/GLS-dependent pyruvate surge offers a promising therapeutic strategy to sensitize tumors to anti-PD-1 immunotherapy.

Programmed Death-Ligand 1 (PD-L1) has been proven to mediate various mechanisms to induce T-cell dysfunction in influenza A virus infection. However, whether PD-L1 similarly contributes to inflammatory responses during influenza B virus (IBV) infection remains unknown. Here we investigated PD-L1 expression patterns during acute IBV infection. A total of 24 IBV-infected patients and 33 healthy controls were recruited and assigned to three age subgroups. Using flow cytometry, we assessed PD-L1 expression on circulating leukocytes and measured the concentrations of plasma cytokines. Our findings demonstrated that IBV infection induced PD-L1 expression on human dendritic cells, T-cells and monocytes. Additionally, IL-2, IL-4, IL-6, IL-10, and TNF concentrations elevated significantly, indicating the occurrence of robust inflammatory responses. Notably, age did not influence PD-L1 expression on peripheral immune cells across the three age subgroups. These results reveal that IBV infection involves the expression of PD-L1 on key immune cell populations, and PD-L1 may serve as a target in the design of new therapeutic strategies for IBV.

Clonal hematopoiesis of indeterminate potential (CHIP), an age-related phenomenon, elevates the risk of hematological malignancies and cardiovascular diseases. JAK2V617F, a key driver mutation in CHIP and myeloproliferative neoplasms (MPNs), has emerged as a novel risk factor for cardiovascular diseases. Atrial fibrillation (AF), the most common arrhythmia, shares a similar onset age with MPNs and is linked to increased risks of thromboembolism and mortality. However, the direct causal relationship and underlying mechanisms between hematopoietic-specific JAK2V617F mutation to AF remain unclear.A retrospective analysis of 1331 patients with myeloid malignancies was performed. Using Fine-Gray multivariable competing risk models adjusted for age, sex, and cardiovascular risk factors, the JAK2V617F mutation was identified as an independent risk factor for new-onset AF (HR=2.20, 95%CI:1.45-3.35, P<0.001). Mediation analysis revealed that IL-1β played a significant mediating role in JAK2V617F-promoted new-onset AF (HR=0.039, 95%CI:0.002-0.080, P=0.042). A hematopoietic-specific JAK2V617F knock-in mouse model (JAK2VF) and littermate wild-type (WT) controls were used. In vivo invasive electrophysiology and ex vivo optical mapping showed that JAK2VF mice had significantly higher AF induction success rates and longer AF duration than WT mice (P<0.05). Regarding atrial structural remodeling: JAK2VF mice exhibited significant left atrial enlargement (P<0.05) and increased atrial fibrosis (P<0.05). Single-cell RNA sequencing of isolated left atrial tissues revealed a significantly activated phenotype of atrial fibroblasts in the JAK2VF group. CellChat analysis demonstrated enhanced macrophage-fibroblast interactions in the JAK2VF group. Further analysis showed that the proportion of CCR2+ macrophages in JAK2VFatrial tissues was significantly increased (P<0.05), along with upregulated expression of NLRP3 inflammasome-related genes Nlrp3 and Il1b (P<0.05). In the mouse model, the number of left atrial macrophages was significantly increased in the JAK2VFgroup, accompanied by NLRP3 inflammasome activation, with a significant co-localization of CD68 and NLRP3. Bone marrow-derived macrophages (BMDMs) from JAK2VFor WT mice were co-cultured with atrial fibroblasts from WT mice. JAK2VFBMDMs significantly promoted the expression of fibrosis-related proteins (collagen 1, α-SMA) in fibroblasts (P<0.05). Meanwhile, JAK2VFBMDMs showed significantly higher expression of key NLRP3 inflammasome components (NLRP3, ASC, Caspase-1) and elevated levels of IL-1β and IL-18 in the supernatant compared to WT BMDMs (all P<0.05). In addition, a retrospective analysis of the JAK2V617F-mutant patient cohort showed that patients receiving the JAK1/2 inhibitor ruxolitinib had a significantly lower incidence of new-onset AF than those not receiving ruxolitinib.In conclusion, this study is the first to demonstrate that the hematopoietic JAK2V617F mutation is an independent risk factor for new-onset AF in both human populations and animal models. Mechanistically, this effect is mediated primarily by JAK2V617F-positive macrophages through activating the NLRP3 inflammasome and releasing cytokines such as IL-1β, thereby driving atrial fibroblast activation and atrial structural remodeling. Clinical data indicate that ruxolitinib may reduce AF risk in JAK2V617F-mutant patients by inhibiting the JAK-STAT pathway and downstream inflammation. Our findings not only elucidate a novel mechanism by which CHIP, particularly JAK2V617F, promotes AF but also provide strong experimental and clinical evidence for targeting inflammatory pathways by using JAK inhibitors in the prevention or treatment of CHIP/MPN-related AF, warranting prospective studies for validation.

BACKGROUND: Patients with thymic epithelial tumors (TETs) frequently show coexistence of various paraneoplastic syndromes, which severely affect their survival. Moreover, there is a lack of effective clinical treatment strategies for patients with unresectable metastatic and recurrent TETs. METHODS: To explore the genetic alterations that play a key role in the pathogenesis of TETs, we analyzed the whole-exome sequencing data from 24 patients diagnosed to have TETs at the First Affiliated Hospital of Nanjing Medical University. RESULTS: Mutated genes in TETs were enriched in several hormone-associated pathways such as insulin secretion; Cushing syndrome; parathyroid hormone; and thyroid hormone, as well as multiple classical tumor-associated pathways, including cAMP, Notch, PI3K-Akt, and WNT signaling pathway. Patients with paraneoplastic syndromes (PNS) exhibited more pronounced alterations in hormone-related pathways. RHPN2 is the most frequently mutated gene in TETs. TETs with RHPN2 mutation showed greater upregulation of the hormone-related signaling pathways such as thyroid hormone and parathyroid hormone as well as a trend toward shorter survival of patients. CONCLUSION: We analyzed the possible role of hormones in TETs on several levels, explored potential links between hormones and other genetic mutations, and found that RHPN2 may be a potentially valuable gene.

Background:Colorectal cancer (CRC) is one of the most prevalent and lethal malignancies worldwide, often characterized by the aberrant activation of multiple signaling pathways. Y-box binding protein 1 (YBX1), a multifunctional regulator of transcription and translation, has been identified as an oncogenic factor in various solid tumors. However, its expression profile and mechanistic role in CRC remain largely unclear. Methods:In this study, integrative bioinformatic analyses were conducted on The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) datasets to assess YBX1 expression and its correlation with CRC progression. Functional assays, including cell proliferation and migration assays, were performed to investigate the role of YBX1 in CRC cells. The impact of YBX1 on the phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) signaling pathway was evaluated, and the effects of the PI3K inhibitor buparlisib (BKM120) on YBX1-driven cellular phenotypes were also tested. Results:YBX1 was found to be significantly upregulated in CRC tissues and was closely associated with the activation of the PI3K/AKT signaling pathway. YBX1 overexpression promoted CRC cell proliferation and migration, whereas knockdown of YBX1 inhibited these processes. Mechanistically, YBX1 was shown to enhance PI3K/AKT signaling activity, promoting malignant phenotypes in CRC. Treatment with BKM120 partially reversed these effects. Additionally, Gene Set Enrichment Analysis (GSEA) identified enrichment of reactive oxygen species (ROS)-related pathways in YBX1-high CRC samples. Conclusions:This study highlights the oncogenic role of YBX1 in CRC and reveals a potential YBX1-PI3K/AKT regulatory axis that may serve as a promising therapeutic target. The findings suggest that targeting this axis could provide a novel strategy for CRC treatment, especially under hypoxic or microenvironmental stress conditions.

AIMS:Mesothelioma (MESO) is a rare and aggressive tumour originating from mesothelial cells, primarily affecting the pleura and peritoneum. Its diagnosis remains challenging due to non-specific clinical, radiological and histopathological features, compounded by morphological diversity and overlapping immunohistochemical profiles with other tumours. Building on our previous finding that GFPT2 is highly expressed in MESO tissues, we evaluated its diagnostic utility in distinguishing MESO from histological mimics. METHODS AND RESULTS:We conducted an immunohistochemical analysis of GFPT2 in 101 MESO cases and 266 histological mimics, including 100 non-small cell lung cancer (NSCLC), 40 high-grade serous ovarian cancer (HGSOC), 6 epithelioid hemangioendothelioma (EHE), 33 solitary fibrous tumour (SFT), 23 aggressive fibromatosis (AF), 6 synovial sarcoma (SS), 7 dedifferentiated liposarcoma (DDLPS), 6 leiomyosarcoma (LMS), 4 malignant peripheral nerve sheath tumour (MPNST), 8 sarcomatoid carcinoma, 20 reactive mesothelial hyperplasia (RMH) and 13 well-differentiated papillary mesothelial tumour (WDPMT) cases. GFPT2 positivity was detected in 85.15% (86/101) of MESO cases, significantly higher than in RMH (0%, 0/20), WDPMT (0%, 0/13), NSCLC (0%, 0/100), HGSOC (0%, 0/40), EHE (16.7%, 1/6), SFT (0%, 0/33), AF (21.74%, 5/23), SS (0%, 0/6), LMS (0%, 0/6), MPNST (25%, 1/4) and sarcomatoid carcinoma (12.5%, 1/8). However, DDLPS (85.7%, 6/7) showed high GFPT2 positivity. CONCLUSIONS:This study demonstrates GFPT2's diagnostic utility in MESO, effectively overcoming tumour heterogeneity challenges. It distinguishes malignant mesothelial lesions from benign/borderline ones (RMH/WDPMT), differentiates epithelioid MESO from epithelioid malignances(NSCLC/HGSOC/EHE) and aids in sarcomatoid MESO versus spindle cell tumours (SFT/AF/SS/LMS/MPNST/sarcomatoid carcinoma), though limited for DDLPS. In summary, GFPT2 is a promising novel antibody demonstrating 85.2% sensitivity and 94.7% specificity.

As hematological tumor patients are surviving long-term, the long-term toxicities of therapeutic regimens have become increasingly evident. The coexistence of two hematological tumors in the same patient is extremely rare and typically shows an aggressive clinical course and unsatisfactory prognosis. In the present case, we describe the case of a 64-year-old man who was admitted to the hospital because of fatigue. Biochemical showed an elevated monoclonal immunoglobulin M (IgM) at 37g/L. Next Generation Sequencing (NGS) analysis revealed MYD88L265p mutation, CXCR4 wild type. In August 2020, he was diagnosed with Waldenström macroglobulinemia (WM) and underwent six cycles of chemotherapy with bendamustine, zanubrutinib, and rituximab. However, he was admitted to the hospital in December 2022 following six-month history of Leukocytosis. Bone marrow (BM) flow cytometry (FCM) showed increased MO1 monocytes. Molecular studies were positive for TET2 mutations. He was finally diagnosed with WM and chronic myelomonocytic leukemia (CMML). Then he accepted hematopoietic stem cell transplantation (HSCT). Unfortunately, after 6 months, the patient died as a consequence of severe pulmonary infection.

Thymoma is often associated with myasthenia gravis (MG). Abnormal lymphocyte differentiation often occurs in the thymoma tumor microenvironment (TME), leading to thymoma-associated autoimmune diseases. Thymoma is closely related to MG, although the underlying mechanisms remain unclear. Patients diagnosed with thymoma were selected and divided into three groups on the basis of MG diagnosis and severity: thymoma alone (T), thymoma-associated MG with mild and moderate clinical symptoms (T + MGL), and severe thymoma-associated MG (T + MGH). Tumor tissue and peripheral blood samples were collected from each group of patients. In the thymoma TME, CD19+ B cells, CD19+CD5+CD1d+ regulatory B cells (Bregs), CD4+ T cells, and CD4+CXCR5+ T follicular helper cells (Tfhs) were localized via multilabel immunofluorescence staining to clarify the relationship between local immune infiltration in the TME and MG severity. Bregs, Tfhs, and other immune cells in the peripheral blood were assessed by flow cytometry. B-cell-enriched regions were detected around blood vessels in the thymoma TME. Breg infiltration in the TME decreased with MG aggravation, whereas the opposite trend was observed for Tfh cells. The Breg/Tfh ratios in the peripheral blood and TME were broadly consistent, and the levels of both types of cells were significantly lower in patients with aggravated MG. Our findings revealed a balance among the Breg/Tfh ratio, immune hyperactivity, and immune tolerance in thymoma-associated MG in both the peripheral blood and the TME. These observations provide new perspectives regarding disease pathogenesis and immunotherapy.

ABSTRACT Background The incidence, outcomes and risk factors for AF in the JAK2V617F‐positive MPN patients are still unknown. Methods The clinical profiles of patients with JAK2V617F‐positive MPN were retrospectively analyzed. Multivariable Cox regression analysis was performed to identify risk factors of AF, thereby developing a risk prediction model. Results A total of 439 patients were included (age 57 [12–87] years; 51.3% male). AF was associated with higher risks of stroke (p = 0.036, HR = 1.987, 95% CI 1.047–3.772) and mortality (p < 0.001, HR = 3.857, 95% CI 1.836–8.103). Multivariable Cox regression showed that TET2 mutation (p = 0.042, HR = 4.361, 95% CI 1.053–18.056) and increased IL‐1β (p = 0.012, HR = 5.476, 95% CI 1.547–28.123) were significant risk factors for AF in patients with JAK2V617F‐positive MPN. Nomograms were constructed, allowing patients to be categorized into high‐ and low‐risk groups. The 10‐year AF‐free survival rate was significantly lower in the high‐risk group (62% vs. 91.7%; log‐rank test: p = 0.002). The validation cohort confirmed that the survival without AF in the high‐risk group was significantly worse than that in the low‐risk group. The use of either interferon‐α or ruxolitinib, was associated with longer AF‐free survival in the high‐risk group (p < 0.05). Conclusion AF was significantly associated with higher risks of stroke and mortality. TET2 mutation and increased IL‐1β were independent risk factors of AF in patients with JAK2V617F‐positive MPN.

Objective: There is no unified standard for the optimal treatment of relapsed/refractory acute myeloid tumors (R/R MN). At present, salvage chemotherapy and hematopoietic stem cell transplantation (HSCT) are mainly used, but most of these patients are not suitable for intensive chemotherapy due to age, physical status, comorbidities and other factors. Cord blood (UCB) has been shown to play a role in hematopoietic reconstruction and graft anti-tumor. The purpose of this study was to explore the clinical efficacy and to analyze the mechanism of cord blood microtransplantation in UNFIT AND R/R MN patients.Methods: A total of 55 patients with ECOG score ≥2 and R/R MN admitted to the Department of Hematology of the Second Hospital of Tianjin Medical University from January 2020 to December 2024 were included in the study (Median age 69 years; Range 26-90 years). They were randomly divided into cord blood microtransplantation group and control group, and all enrolled patients underwent 1 cycle of salvage chemotherapy. The umbilical cord blood micrograft (UCB) group was infused with UCB as adjuvant therapy 1 day after the end of chemotherapy (Total number of MNC (14.45×109, 2-21.81×109); total CD34+ count (3.90×107, 0.93-8.79×107); Percentage of NK cells (1.49%, 0.22-2.48%). Clinical efficacy analysis mainly assesses the overall complete response rate (CR) or complete response with incomplete haematological recovery (CRi). Overall survival (OS), event-free survival (EFS), minimal residual disease (MRD) negativity rate, and blood cell recovery time were used as secondary evaluation indicators. The number of immune cells and cytokine levels were analyzed 28 days after treatment to evaluate the micro-changes in immunity.Results: The median follow-up was 178.5 (1-1505) days. Compared with the control group, 17 patients in the UCB group achieved CR and PR (45.1% vs 33.3%, P=0.026) ORR. The median event-free survival (EFS) was better (P=0.019), the median overall survival was better (P=0.026), the median overall survival was better (P=0.039) in the patients with NR (P=0.039), and the patients with TP53 mutation had better overall survival in the umbilical cord blood adjuvant group (P=0.04). 11 patients in the UCB group achieved MRD negativity (P= 0.026). The median time to neutrophil, hemoglobin and platelet recovery was 22 days (P=0.008), 21 days (P=0.002), and 22 days (P=0.006), respectively. The median recovery time of neutrophils, red eggs and platelets was 22 days (P=0.008), 21 days (P=0.002) and 22 days (P=0.006). No acute graft-versus-host disease (aGVHD) or toxicity-related deaths occurred in any of the patients. At 28 days after microtransplantation, the number of CD3+ T cells, CD8+ effector memory T cells, NK cells and NK Mature cells increased compared with those before reinfusion (P=0.005, P=0.05, P=0.05, P=0.05), and the number of CD19+ B cells decreased (P=0.05). INF-γ level (P=0.05) and TNF-α level increased (P=0.03) at 28 days after microtransplantation.Conclusion: Umbilical cord blood microtransplantation can improve OS and EFS in patients with R/R myeloid tumors who are not suitable for intensive chemotherapy, and reduce the time of hematopoietic reconstruction after chemotherapy. The increase in the proportion of NK and T cells after cord blood microtransplantation improves chemotherapy outcomes by inducing microGVL effects.

Background:Thymic epithelial tumors (TETs), categorized predominantly as thymoma (T) or thymic carcinoma (TC), face a challenging prognosis and limited treatment options. Although chemotherapy remains the established treatment for advanced TETs, its responses tend to be short-lived. The emergence of immunotherapy, particularly programmed cell death-1 (PD-1) and programmed death ligand-1 inhibitors (PD-L1), is increasingly being regarded as a promising new treatment option for various malignancies. Methods:Herein, we present a case series of eight patients with TETs who received tislelizumab treatment at Jiangsu Provincial Hospital between 2021 and 2023. All cases were histologically confirmed as either thymoma or thymic carcinoma. Among these eight cases, six patients (5 thymic carcinomas [TC] and 1 thymoma [T]) received tislelizumab in combination with chemotherapy following multiple cycles of prior chemotherapy without achieving significant therapeutic response. Two TC patients were administered this combination regimen as first-line treatment. Following the initiation of immunotherapy, patients received tislelizumab at a dose of 200 mg every three weeks until disease progression or the occurrence of unacceptable toxicity. Treatment response was assessed by the investigators according to the Response Evaluation Criteria in Solid Tumors (RECIST) version 1.1 guidelines. Results:The 8 patients described had a median age of 59 years (range, 47-72). During the course of immunotherapy, five patients (62.5%) achieved partial response, and notably, even after transitioning to maintenance therapy with tislelizumab, the lesions continued to shrink, with the longest sustained partial response lasting over 2 years. Three patient (37.5%) experienced stable disease as their best response to immunotherapy. Among all these patients, three patients (37.5%) demonstrated initial efficacy but subsequently exhibited progressive disease (median progression-free survival of 14 months). All patients are still being followed up, with the longest PFS extending to 31 months. Notably, five of the eight patients underwent PD-L1 testing and were all found to be negative. Despite this, no immune-related Grade 3-5 adverse events (AEs) were reported and all AEs were manageable with supportive measures. Grade 1-2 AEs were adrenal insufficiency (n=1), thyroid dysfunction (n=1), and pneumonia (n=1). Conclusions:Our study findings suggest that the combination of immunotherapy and chemotherapy yields durable clinical responses in patients with TETs, suggesting its potential as a safe and effective first-line treatment strategy for advanced TETs. Notably, the therapeutic benefits of chemo-immunotherapy appear to extend beyond patients with high PD-L1 expression (≥50%), indicating that this treatment approach may not be strictly limited to individuals with elevated PD-L1 levels.

Background: Diffuse large B-cell lymphoma (DLBCL) is predominant subtype of non-Hodgkin lymphoma and can be effectively treated. Nevertheless, a subset of patients experiences refractory or relapsed disease, highlighting the need for new therapeutic strategies. Methods: Depmap database based on CRISPR/Cas9 knock out analysis was employed to identify the essential gene SH3GL1, which encodes endophilin A2, as crucial for the proliferation and survival of DLBCL cells. Immunohistochemistry (IHC) staining was performed on the 126 paraffin-embedded clinical DLBCL samples to investigate the association between SH3GL1 expression levels and the prognosis. To investigate the specific mechanism modulated by SH3GL1 in the progression of DLBCL, an integrative approach was employed. This approach combined high-throughput sequencing technologies, such as Deep-DIA and LC-MS, with functional validation techniques, including CRISPR/Cas9 gene editing, xenograft models, and molecular pathway analyses. Results: Our study found that high expression levels of SH3GL1 correlate with poor prognosis in a cohort of 126 newly diagnosed DLBCL patients, underscoring its significance in disease progression. Mechanistically, we found that SH3GL1 deficiency triggers ferritin heavy chain 1 (FTH1)-mediated ferroptosis, specifically ferritinophagy-induced ferroptosis, in DLBCL cells. Additionally, high expression of SH3GL1 suppresses doxorubicin-induced ferroptosis. Cancer cells' resistance to conventional therapies is associated with increased sensitivity to ferroptosis. Conclusions: These findings emphasise SH3GL1 as a promising prognostic biomarker and a potential therapeutic target in DLBCL, offering new avenues for treatment strategies aimed at overcoming drug resistance and improving patients' outcomes. Key points Elevated SH3GL1 expression in DLBCL patients was associated with a negative prognosis. SH3GL1 plays a crucial role in promoting DLBCL cell survival through the regulation of FTH1-mediated ferroptosis and doxorubicin resistance.

Abstract Myelodysplastic syndromes/tumors (MDS) are a highly heterogeneous group of myeloid malignancies. It has been reported that 1.38%~20% of MDS patients were accompanied by T cell large granulosa lymphocyte proliferation (LGLP) at the initial diagnosis, and is different from that of MDS combined with T-LGLL in mechanism. Komrokji et al. suggested that MDS-LGLP may be a benign expansion phenomenon caused by innate immunity due to tumor cell activation (Komrokji RS, et al. Leukemia, 2020). While enhanced T cytotoxicity inhibits bone marrow hematopoiesis and causes a decrease in normal blood cells, it also shows higher killing activity against tumor cells, suggesting that CTLs play a role in inhibiting malignant clones in early MDS (Tentori CA, et al. Hemasphere, 2024).In this study, we first performed a retrospective analysis of clinical and laboratory data from 22 MDS-LGLP patients and 66 age-, sex-, and diagnosis-matched MDS patients. The results showed that patients in the MDS-LGLP group had better survival outcomes (P=0.02). The Th/Tc, proportion of CD4+ Tcm cells, total NK cells, mature NK cells, pDCs, and mDCs in the MDS-LGLP group was higher respectively (P<0.05, P<0.01, P<0.01, P<0.05, P<0.01, P<0.001). The proportion of Th17 cells, and CD8+ Tcm cells in the MDS group was higher (P<0.05, P<0.05). In terms of cytokine levels, the MDS-LGLP group showed significantly reduced levels of immune suppressive factors associated with immunomodulation and tumor immune escape (IL-10, TGF-β, IL-6; P<0.01, P<0.001, P<0.001). It suggests that the higher overall survival rate of MDS-LGLP patients may be related to the enhanced anti-tumor immune microenvironment.Given that the presence of LGLP may delay the progression of MDS disease, we set to explore the molecular mechanism of MDS-LGLP occurrence at the single-cell sequencing level. We collected bone marrow mononuclear cells from 3 MDS-LGLP and 3 MDS patients for single-cell RNA sequencing (scRNAseq) and analyzed the results, distinguished different cell subsets by cell surface markers, observed the differences in hematopoietic stem/progenitor cells, lymphoid subsets and TCR clones, and tracked the differences in cell differentiation and development. Through cell differentiation trajectory analysis, we found that hematopoietic progenitor cells (HSPCs) of MDS-LGLP patients had lower tumor stem cell characteristics, and early lymphatic cells showed more terminal differentiation characteristics. The KEGG apoptosis-related pathway score of HSPCs in MDS-LGLP patients was higher than that in MDS patients, consistent with the results of cell differentiation trajectory analysis, suggesting that HSPCs of MDS patients had higher tumor stem cell characteristics and apoptosis arrest. Transcriptome combined with V(D)J sequencing showed that the TCR CDR3 distribution of MDS-LGLP and MDS patients was similar, indicating that MDS-LGLP may be a reactive proliferation of T cells under tumor proliferation stimulation, and the TCR cloning pattern of MDS-LGLP showed a significant trend of shifting from polyclonal to oligoclonal.In addition, DEGs analysis of lymphocyte subsets in MDS-LGLP and MDS groups showed that TMSB10 was significantly highly expressed in MDS patients than MDS-LGLP patients, and was widely positively correlated with immunosuppressive genes such as PD-1, TIM-3 (P＜0.001, P＜0.001 respectively). Immune infiltration analysis showed that the expression level of TMSB10 was significantly positively correlated with the degree of CD8+ T cell infiltration (P＜0.05). GSEA analysis showed that high expression of TMSB10 in the MDS group could upregulate NFAT/NF-κB and IL-10 signaling pathways, which has been shown to be closely related to T cell depletion and anti-tumor immune tolerance.In summary, our comprehensive analysis of clinical data, laboratory characteristics and scRNAseq data showed that in MDS-LGLP patients, cytotoxic T lymphocytes and NK cells functions as to recognize and kill tumor cells by self-proliferation, and low expression of TMSB10 in lymphocytes may be a characteristic change marker of the disease state. The reduction of TMSB10 level may improve the immune tolerance status of T cells, enhance anti-tumor immunity, and render these patients a better survival outcomes. Further experiments are underway to fully understand the underlying mechanisms and potential clinical treatments for this disease.

Background:Angiogenesis inhibition represents a key therapeutic strategy in oncology, with apatinib showing significant efficacy for advanced gastric cancer (AGC) therapy. Currently, no biomarkers can reliably predict which AGC patients will benefit most from apatinib. The purpose of this study was to evaluate apatinib plus tegafur gimeracil oteracil potassium capsule (S-1) as a second-line therapy in AGC and to investigate prognostic biomarkers associated with outcomes. Methods:All 60 patients received apatinib (250 or 500 mg once daily) in combination with S-1 (40 mg/m2, administered twice daily from day 1 to day 14). Treatment was administered in 28-day cycles, with each course encompassing two such cycles. Multiparameter flow cytometry was used to analyze the subset distribution and immunophenotypes of T cells and natural killer (NK) cells in peripheral blood mononuclear cell samples from AGC patients. Results:Among all 60 evaluable patients, no complete responses were observed. Partial responses were observed in 33.3% (20/60) of patients (95% confidence interval (CI): 21.1-45.6%), stable disease was observed in 38.3% (23/60) of patients (95% CI: 25.7-51.0%), while progressive disease occurred in 28.3% (95% CI: 16.6-40.1%). Notably, patients with NK cell proportions greater than 17% experienced longer progression-free survival (PFS) and overall survival (OS) than those with lower proportions. Conclusions:Apatinib combined with S-1 exhibited favorable tolerability and promising clinical activity as a second-line option for AGC patients. Importantly, we identified peripheral blood NK cell proportion as a potential predictor of treatment response, where a low NK cell proportion correlated with poorer outcomes.

Hepatocellular carcinoma (HCC) is a malignant tumour with high prevalence and mortality rate worldwide. Metabolic reprogramming of cancer cells may be a major factor in the process of this disease. Glucose transporter proteins (GLUTs) are members of the major facilitator superfamily of membrane transporters, playing a pivotal role in the metabolic reprogramming and tumour progression in HCC. This review discusses the advances in the study of GLUTs in HCC, including the expression patterns, functions and possibilities of GLUTs. In HCC, the expression levels of GLUTs are closely associated with tumour aggressiveness, metabolic reprogramming and prognosis. A series of inhibitors have been demonstrated efficacy in inhibiting HCC cell growth and glucose uptake in in vitro and in vivo models. These inhibitors offer a novel approach to HCC treatment by reducing the glucose metabolism of tumour cells, thereby impeding tumour growth, and concurrently enhancing the sensitivity to chemotherapeutic agents. This reminds us of the urgent need to elucidate GLUTs' roles in HCC and to determine the most effective ways to translate these findings into clinical practice.