Compared with anti-citrullinated protein autoantibody (ACPA)-positive RA, the etiology of ACPA-negative RA remains unclear, posing significant challenges to diagnosis. Recently, evidence has emerged that ACPA-negative RA is characterized by metabolic reprogramming, as well as dysregulated lipid metabolism. The remnant cholesterol inflammation index (RCII) is a comprehensive index of lipid burden and inflammation; this study evaluated the predictive value of RCII as a novel biomarker for ACPA-negative RA. In the Beijing Hospital (BJH) cohort, we first compared RCII across various autoimmune diseases and undifferentiated inflammatory arthritis versus healthy controls (HCs). Subsequently, we assessed the independent association between RCII and ACPA-negative RA in the BJH cohort and UK Biobank (UKB) cohort using multivariable logistic regression, adjusting for confounders including age, sex, BMI, and cardiovascular comorbidities. Diagnostic performance was evaluated via receiver operating characteristic (ROC) analysis, and optimal diagnostic RCII intervals were defined using a stratified 5-fold cross-validation framework. Furthermore, the possible underlying mechanisms were investigated using untargeted serum lipidomics in a matched subset. In the BJH cohort, RCII levels were significantly elevated in ACPA-negative RA, ACPA-positive RA, undifferentiated inflammatory arthritis, dermatomyositis, and systemic lupus erythematosus compared with HCs, with the highest levels observed in ACPA-negative RA. Dual-cohort analysis confirmed elevated RCII in ACPA-negative RA, with independent associations (BJH: OR = 26.11; UKB: OR = 5.59) even after adjusting for age, sex, BMI, and cardiovascular disease (CVD). In the ROC analysis, RCII demonstrated relatively high diagnostic accuracy, achieving an area under the curve (AUC) of 0.929 in the BJH cohort and 0.785 in the UKB cohort. Cross-validation established stable diagnostic intervals, effectively minimizing the indeterminate decision zone, or “gray zone” to less than 10

ObjectivesVEXAS syndrome is a recently characterized hemato-inflammatory disorder caused by somatic mutations in the X-linked UBA1 gene in hematopoietic cells, which remains poorly characterized in Chinese populations. This study aims to address this gap.MethodsWe retrospectively analyzed 4512 consecutive patients with hematologic abnormalities at a Chinese academic hospital between June 2023 and July 2024, identifying 16 male VEXAS patients (median age 61.5 years, range 30-74).ResultsAll patients presented with anemia and lymphopenia, with or without neutropenia and thrombocytopenia. Diagnoses included CCUS, MDS, MGUS, as well as novel phenotypes of primary myelofibrosis and a hemolysis-like disorder. Most patients (11/16, 69%) exhibited constitutional symptoms and typical autoinflammation-associated multiorgan involvement, including skin lesions, ear chondritis, pulmonary infiltration, and deep vein thrombosis, etc. Hypercellular bone marrow was commonly seen in core biopsies and 11 patients (68.8%) exhibited typical vacuoles in myeloid and erythroid progenitors. Canonical UBA1 pathogenic variants were detected in 81.3% (13/16) of patients, with p.M41V being the dominant mutation. Three infrequent variants were also identified: c.118-1G>C, p.S56P, and p.S621C. Corticosteroids and immunosuppressants commonly provided symptomatic relief, while variable hematologic responses were achieved with androgens and erythropoiesis-stimulating agents.ConclusionsAs a relatively large cohort of VEXAS syndrome characterizing Chinese patients, our findings demonstrate that VEXAS should be considered in those with cytopenia, regardless of systemic symptoms or multiorgan involvement. Increased awareness among hematologists is critical to facilitate early diagnosis via UBA1 testing. This can prevent unnecessary diagnostic procedures and guide appropriate treatment, including consideration of pre-emptive stem cell transplantation.

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Inducing adult cardiomyocyte proliferation to repair the infarcted heart remains a major therapeutic challenge. While metabolic reprogramming is known to drive regeneration, the specific organelle-level mechanisms governing this process, particularly the crosstalk between mitochondria and lipid droplets (LDs), remain elusive. Here, we identify Heat Shock Cognate 71 kDa Protein (Hsc70) as a critical physiological "metabolic brake" that maintains adult cardiomyocytes in a terminally differentiated state and suppresses cell cycle re-entry by tethering mitochondria to LDs via Mitofusin 2 (Mfn2). Using Ginsenoside Rb2, a bioactive small molecule identified from a clinically effective formula (Shuangshen Ningxin), we demonstrate that Rb2 directly binds to Hsc70 (KD ≈ 32 µM) and disrupts the Hsc70-Mfn2 interaction. This disruption pharmacologically uncouples the remaining mitochondria-LD contacts to f release this physiological barrier, restores metabolic homeostasis, and reactivates cardiomyocyte proliferation in myocardial infarction (MI) rats. Crucially, these regenerative effects were abrogated by AAV9-mediated Hsc70 overexpression, confirming Hsc70 as the non-redundant therapeutic target. Furthermore, a retrospective analysis of 60 patients treated with the Rb2-containing intervention showed significantly improved cardiac outcomes, highlighting the broad cardioprotective and clinical utility of this therapeutic strategy. Our findings reveal a fundamental mechanism linking organelle dynamics to tissue regeneration and highlight Hsc70 as a druggable target for heart failure treatment.

The accumulation of depolarized mitochondria commits T cells to exhaustion1-3, yet the precise mechanism remains unclear. Here we find that exhausted CD8+ T cells increase proteasome activity owing to the accumulation of depolarized mitochondria, which drives the selective degradation of mitochondrial proteins and the release of regulatory haem through haemoprotein breakdown. In turn, increased regulatory haem disrupts BACH2-mediated transcriptional regulation, thereby exacerbating T cell exhaustion and compromising stemness-like properties. Inhibition of nuclear import of regulatory haem prevents BACH2 degradation and enhances the anti-tumour efficacy of antigen-specific T cells. We find that the therapeutic efficacy of human CD19+ chimeric antigen receptor (CAR)-T cells in patients with B cell acute lymphoblastic leukaemia negatively correlates with the proteasome gene signature in their CAR-T cells. Manufacturing CAR-T cells in the presence of bortezomib, an FDA-approved proteasome inhibitor, prevents T cell exhaustion and improves therapeutic efficacy. Our findings identify a proteasome-guided haem signalling axis, governed by mitochondrial integrity, as a regulator of CD8+ T cell exhaustion and propose innovative therapeutic strategies that exploit this pathway to optimize adoptive cellular immunotherapy.

There is limited information on the uncontrolled status and burden of asthma among patients using inhaled corticosteroids (ICS) combined or not with long-acting β2-agonists (LABA) in China. This study aims to characterize the disease burden among these patients and identify risk factors for unfavorable outcomes. This retrospective observational study utilized data from a regional Electronic Medical Record (EMR) database in China. Patients (≥ 12 years) with asthma having at least two prescriptions for medium-to-high dose ICS or ICS/LABA within any 6 months were identified from the Shanghai Medical Database between 2016 and 2020. Data on characteristics and treatment patterns 1-year pre-index (baseline) and disease burden 1-year post-index (follow-up) were obtained from the database. Uncontrolled asthma was defined based on exacerbations and specific treatment. A multivariate logistic regression model was used to identify risk factors associated with the follow-up adverse outcomes. A total of 35,900 patients with asthma receiving medium-to-high dose ICS or ICS/LABA were included. Of these, 10,842 (30.2

The bone marrow (BM) niche serves as a critical protective environment for leukemia cells, particularly chemo-resistant leukemia cells, and plays a central role in driving therapeutic resistance and disease relapse in acute myeloid leukemia (AML). This specialized microenvironment not only promotes leukemia cell survival, but also inhibits T cell infiltration, which serves as a major obstacle to the effectiveness of CAR-T therapy in myeloid malignancies. To overcome this limitation, we targeted Rac1 GTPase, a central regulator of cytoskeletal dynamics that controls membrane protrusion and migration, by engineering primary human T cells and CD33 CAR-T cells to express constitutively active Rac1 (Rac1V12). Our results demonstrated that active Rac1 enhanced the migration of T cells and CD33 CAR-T cells and promoted their residence in the BM in vivo. Furthermore, CD33 CAR-T cells expressing Rac1V12 displayed enhanced cytotoxicity against leukemia cells in vitro, as demonstrated by transwell migration-dependent killing assays. Crucially, these engineered CAR-T cells achieved superior robust suppression of leukemia in vivo and significantly prolonged survival in xenograft models. Mechanistically, Rac1V12 CD33 CAR-T cells in the BM demonstrated enhanced immunological memory phenotype and lower tonic signaling, a combination that promotes T cell persistence and enhances anti-tumor efficacy in vivo. Our data suggest that active Rac1-engineered CD33 CAR-T cells represent a novel strategy for targeting BM leukemia cells, with the potential to eradicate AML cells.

Carthamus tinctorius L. is widely used for cardiovascular and cerebrovascular diseases. Hydroxysafflor yellow A (HSYA), its major quinochalcone glycoside, is effective against myocardial ischaemia/reperfusion injury (MI/RI), while other compounds' roles remain unclear. This study established chemical fingerprints of 9 C. tinctorius populations via ultra-performance liquid chromatography, evaluated cardiomyocyte protective effects using Cell Counting Kit-8 assays and performed spectrum-effect analysis integrating grey correlation analysis and partial least-square (PLSR), combined with compound identification via ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry. Seven potential active components were screened: HSYA, kaempferol-3-O-β-sophorose, saffloquinoside A, roseoside, quercetin-3,7-di-O-β-D-glucoside, 6-hydroxykaempferol-3,6-di-O-β-D-glucoside, saffloquinoside C. These may be key anti-MI/RI substances in C. tinctorius, with specific pharmacological mechanisms to be further verified, providing a reference for exploring its representative active components.

To develop and validate a prognostic model based on palmitoylation‑related genes for risk stratification in acute myeloid leukemia (AML), and to explore its associations with the immune microenvironment, drug sensitivity, and single‑cell expression patterns. A five‑gene prognostic signature was constructed using univariate Cox regression and LASSO‑Cox analyses in the TCGA‑LAML training cohort (n = 131) and validated in an independent GEO cohort (GSE71014, n = 104). Risk scores were calculated, and patients were divided into high‑ and low‑risk groups. Overall survival (OS) was compared by Kaplan‑Meier curves, and model performance was evaluated by time‑dependent ROC. Immune cell infiltration, immune checkpoint expression, and drug sensitivity (predicted IC50) were analyzed. Single‑cell RNA‑seq data (GSE116256) were used to examine cell‑type specific expression of the model genes. The five‑gene model (LYPLA2, ZDHHC11, ZDHHC7, ABHD6, ABHD17C) significantly distinguished high‑risk patients with poorer OS in both training (P < 0.001) and validation (P < 0.001) sets, with AUCs of 0.72, 0.73, 0.75 (training) and 0.67, 0.70, 0.60 (validation) for 1‑, 3‑, and 5‑year survival. Multivariate Cox regression confirmed independent prognostic value (HR = 2.78, P < 0.001). High‑risk patients showed increased regulatory T cells, activated dendritic cells, and follicular helper T cells, along with positive correlations with PD‑1, TIM‑3, LAG3, and VISTA expression. Ninety‑eight compounds with differential predicted IC50 values were identified. Single‑cell analysis revealed LYPLA2 and ZDHHC7 enrichment in monocytes/macrophages and dendritic cells. The five‑gene palmitoylation‑associated prognostic model effectively stratifies AML patients by risk, reflects immune microenvironment dysregulation, and offers potential guidance for personalized therapy.

BACKGROUND:Aspirin is frequently employed for the prevention of cardiovascular events, but its clinical utility is hindered by the risk of severe gastrointestinal injury when taken orally. Fufanglongxuejie capsules (FFLXJ), a Chinese patent medicine known for promoting wound healing and alleviating congestion and pain, may offer a promising solution to this clinical challenge. METHODS:Using network pharmacology, candidate targets of FFLXJ, gastrointestinal disorders, intersection targets, and associated signaling pathways were examined. Prior to the creation of myocardial ischemia-reperfusion (MI/R) models, male Sprague-Dawley (SD) rats were orally administered FFLXJ and/or aspirin for a consecutive month. Subsequently, serum motilin (MTL), gastrin (GAS), HE staining, transmission electron microscopy analysis, and western blot analysis were performed on the blood samples or gastric tissues. Molecular docking analysis on core targets and relative compounds was conducted using Discovery Studio software. The expressions of core targets were verified by Western blot. RESULTS:Compared with aspirin-treated MI/R rats, FFLXJ restored downregulated serum MTL and GAS levels and lessened aspirin-induced gastrointestinal lesions. Network pharmacology research revealed that the top 4 core targets were TNF, IL-10, PTGS2, and VEGFA. In MI/R rats, aspirin treatment markedly increased the level of stomach IL-10, while FFLXJ administration decreased the expression of PTGS2 and IL-10 compared with aspirin-treated group. CONCLUSION:Oral aspirin harmed the gastrointestinal mucosa in MI/R rats; however, FFLXJ was able to mitigate the damage. The protective property of FFLXJ was related to the regulation of inflammation.

Endoplasmic reticulum stress (ERS) and apoptosis are hallmark pathological features of myocardial ischemia-reperfusion injury (MIRI). Lamin A/C, a nuclear lamina protein associated with cardiac disorders, has been implicated in ERS and apoptosis regulation, yet its role in MIRI remains elusive. Meanwhile, P4HB, an ERS-associated chaperone, may interact with lamin A/C to modulate MIRI progression. We hypothesized that lamin A/C interacts with P4HB to modulate MIRI progression. We assessed the distribution of P4HB and lamin A/C in MIRI SD rat hearts and oxygen-glucose deprivation/reoxygenation (OGD/R)-treated primary neonatal rat cardiomyocytes (PNRCMs). By knocking down lamin A/C expression, we examined the impact of lamin A/C on P4HB distribution, ERS and apoptosis induced by OGD/R modeling. STRING network analysis and protein-protein docking were employed to predict the structural basis of lamin A/C/P4HB interaction. Our results demonstrated OGD/R treatment triggered P4HB nuclear envelope translocation, ERS activation, and apoptosis in PNRCMs, while those effects were attenuated by lamin A/C knockdown. The physical interaction between lamin A/C and P4HB in cardiac tissue was observed under both normal and MIRI conditions. The P4HB-lamin A/C interaction may be dynamically modulated by calreticulin. Collectively, our findings propose that lamin A/C regulates P4HB to mitigate OGD/R-induced ERS and apoptosis, potentially through a calreticulin-mediated dynamic mechanism.

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Immune-related adverse events (irAEs) are associated with anti-PD-1/PD-L1 therapy in gastrointestinal cancers. Altered phenotypes and frequencies of T cell subsets play a critical role in irAEs. This study characterized the Tfh/Treg balance in irAEs patients and assessed the efficacy of LDIL-2 therapy in mouse models. A prospective study evaluating the immunological characteristics of 26 gastrointestinal cancer patients through both cellular and transcriptomic analyses before and after anti-PD-1/PD-L1 therapy was performed. Patients were categorized into two groups: the AE group (patients who developed irAEs during the follow-up period) and the NAE group (patients who did not). Meanwhile, MRL/MpJ-Faslpr mice, a lupus mouse model, and tumor-bearing C57BL/6 J mice were used to evaluate the efficacy of LDIL-2 in treating anti-PD-L1 induced irAEs and its impact on antitumor effectiveness. Our study discovered that in AE patients, Tfh-related genes were significantly upregulated during the early stages of anti-PD-1/PD-L1 therapy (IL-21: p < 0.01; PDCD1: p < 0.05), whereas the upregulation of Treg-related genes was less pronounced compared to Tfh-related genes. This distinction was not observed in NAE patients. Flow cytometry results further confirmed a substantial increase in the Tfh/Treg ratio in AE patients during the early stages of therapy (p < 0.001), a phenomenon absent in NAE patients. In MRL/MpJ-Faslpr mice, anti-PD-L1 therapy induced a shift in the Tfh/Treg ratio, along with severe organ inflammatory lesions and elevated anti-dsDNA antibody levels. In contrast, LDIL-2 treated mice maintained a balanced Tfh/Treg ratio, exhibiting significantly fewer organ inflammatory lesions and reduced anti-dsDNA production. Furthermore, in tumor-bearing mice, LDIL-2 did not impair the antitumor efficacy of anti-PD-L1 therapy. Our study underscores the importance of Tfh/Treg balance in the development of irAEs and demonstrates the potential of LDIL-2 as a therapeutic option for irAEs, providing a promising alternative to traditional immunosuppressive therapies for managing irAEs.

Acute myeloid leukemia (AML) is a hematological malignancy with a high mortality rate and heterogeneous prognosis. Traditional risk stratification is based on the genetic classification in the 2022 guidelines of the European Leukemia Net. However, the risks of some patients remain unclear, and other prognostic assessment methods are required to improve the risk assessment of these patients. Apoptosis-related genes (ARGs) play critical roles in regulating the survival and drug resistance of AML cells. Therefore, we collected gene expression and clinical data from patients with AML from The Cancer Genome Atlas Acute Myeloid Leukemia (TCGA-LAML) datasets to develop a risk assessment model based on 5 ARGs. Using the least absolute shrinkage and selection operator Cox regression (LASSO-Cox) model, we identified 5 key ARGs (DDIT4, HSP90B1, ENO1, SOD1, and SLC7A11) and constructed a 5-ARG prognostic model. Using this model, we successfully stratified patients in both TCGA-LAML training and independent external validation cohorts, with high-risk patients consistently exhibiting significantly poorer clinical outcomes. In addition, high-risk patients exhibited significant enrichment in pathways related to TP53 dysfunction, mechanistic target of rapamycin complex 1 (mTORC1) signaling activation, and pro-inflammatory responses, which were closely correlated with NPM1c-FLT3 co-mutations. Decitabine, sunitinib, and MK-1775 were identified as potential therapeutic agents. In summary, we established a 5-ARG prognostic model that may facilitate risk stratification and inform therapeutic decision-making in AML.

Abstract Introduction In systemic lupus erythematosus (SLE), aberrant B cell activation is fueled by elevated oxidative phosphorylation (OXPHOS), yet checkpoints restraining this metabolic hyperactivity remain undefined. We identify mitochondrial carrier homolog 2 (MTCH2),as a critical metabolic gatekeeper that limits OXPHOS-driven autoreactive B cell expansion and maintains germinal center (GC) tolerance in autoimmunity. Methods We analyzed metabolic profiles of B cells from SLE patients and healthy controls using scRNA-seq. B cell-specific MTCH2-deficient mice (Mb1-Cre; Mtch2fl/fl) were subjected to pristane-induced lupus. GC responses, BCR repertoire, and somatic hypermutation (SHM) were assessed by flow cytometry, and BCR sequencing. Mechanistic studies employed immunoprecipitation and GTPase activity to define the MTCH2—FUNDC2 axis, and a high-throughput screen identified a pharmacological agonist. Results B cells from SLE patients exhibited an OXPHOS signature and concomitantly downregulated MTCH2. B cell-specific MTCH2 deletion in mice exacerbated lupus, with expanded GCs, elevated autoantibodies. Sc-metabolic profiling revealed profound metabolic reprogramming across B cell subsets, with dark-zone GC B cells exhibiting the highest OXPHOS activity. This metabolic rewiring skewed the BCR, promoting the clonal expansion of autoreactive cells with excessive SHM. Mechanistically, MTCH2 governed mitochondrial dynamics via the FUNDC2—MFN1/2 axis which led to dysregulated mitochondrial fusion and B cell activation threshold. Pharmacological upregulation of MTCH2 by agonist restored mitochondrial homeostasis and ameliorated lupus in vivo. Conclusion We establish MTCH2 as a pivotal metabolic checkpoint that restrains autoreactive B cell expansion. Loss of MTCH2 disrupts GC tolerance, driving clonal dominance of autoreactive B cells and accelerating systemic autoimmunity. Targeting the MTCH2 axis represents a clinically translatable strategy to reinstate metabolic control and restore immune tolerance in SLE. Funding Source National Natural Science Foundation of China ( 82230060, 32141004, 32430036, 32441093) Topic Categories Basic Autoimmunity (BA)