Xuefeng Wang

Glucose-6-phosphate dehydrogenase (G6PD), the committed enzyme of the pentose phosphate pathway, plays a pivotal role in glucose metabolism, which is integral to platelet biology. Although G6PD deficiency is a prevalent enzyme defect worldwide, the specific role of G6PD itself in platelets remains unclear. Herein, we discover a previously unrecognized phenotype in both human patients and murine models, demonstrating that G6PD deficiency leads to elevated platelet counts through enhanced proplatelet formation (PPF). Mechanistically, we reveal a non-canonical function of G6PD in regulating actin dynamics via its interaction with an actin-binding protein, tropomodulin-3 (TMOD3). The G6PD-TMOD3 interaction, which is concentration-dependently enhanced by nicotinamide adenine dinucleotide phosphate (NADP ⁺ ), can limit actin rearrangement, thereby inhibiting PPF. Furthermore, we develop a G6PD-derived peptide, GC2P2, which potentiates the G6PD-TMOD3 interaction and significantly suppresses thrombocytopoiesis both in vitro and in vivo. In conclusion, our findings suggest that G6PD deficiency facilitates platelet biogenesis by altering TMOD3-mediated actin dynamics. The study provides new insights into thrombopoiesis and offers potential strategies for modulating platelet production.

BACKGROUND: Apo (apolipoprotein) AI glycation has been implicated in HDL (high-density lipoprotein) dysfunction, but its site-resolved landscape, clinical relevance, relationship with diabetic atherosclerosis, and underlying mechanisms remain incompletely defined. METHODS: We performed unbiased site-resolved glycation proteomics in 860 patients with type 2 diabetes and coronary atherosclerosis (CAS) and 294 controls with type 2 diabetes without CAS to define the plasma apo AI glycation landscape. Apo AI glycation signatures associated with diabetic CAS were identified using integrated unsupervised and supervised analyses. An Apo AI Glycation Index was developed using least absolute shrinkage and selection operator regression and evaluated in training, testing, and independent validation cohorts. To assess functional relevance, a glycation-resistant apo AI mutant, cross-linked apo AI (apo AI _CL ), was engineered and tested under glycation conditions using surface plasmon resonance, in vitro and in vivo reverse cholesterol transport assays, HDL remodeling analyses, and atherosclerosis models. RNA sequencing, macrophage-specific knockout models, receptor-binding assays, signaling inhibition, and cholesterol efflux rescue experiments were used to investigate the underlying macrophage pathway. RESULTS: Apo AI glycation patterns differed between patients with type 2 diabetes with and without CAS, with K96 and K106/107 emerging as prominent disease-associated glycation sites. The Apo AI Glycation Index was independently associated with CAS and coronary artery disease in patients with type 2 diabetes and was inversely associated with HDL-mediated reverse cholesterol transport and lecithin-cholesterol acyltransferase activity. Compared with native apo AI, apo AI _CL showed reduced key-site glycation, preserved structural stability, enhanced binding affinity to lecithin–cholesterol acyltransferase, and improved HDL-mediated reverse cholesterol transport under glycation conditions. In diabetic mouse models, apo AI _CL attenuated HDL dysfunction and atherosclerotic lesion formation compared with glycated apo AI. These data support a candidate mechanism in which glycated apo AI enhances RAGE (receptor for advanced glycation end products) interaction and activates ERK1/2 (extracellular signal-regulated kinase 1/2)–NF-κB (nuclear factor κB)/p65 signaling, leading to upregulation of NR2C2 (nuclear receptor subfamily 2 group C member 2). LXRα (liver X receptor α) activation and macrophage NR2C2 deficiency restored cholesterol efflux in glycated apo AI–treated macrophages. CONCLUSIONS: This study provides a comprehensive site-resolved map of apo AI glycation in patients with diabetes and identifies the Apo AI Glycation Index as a glycation signature associated with diabetic CAS and impaired HDL function. Experimental data support a mechanism in which site-specific apo AI glycation contributes to HDL dysfunction and macrophage cholesterol efflux impairment through RAGE-associated NR2C2–LXRα signaling. Glycation-resistant apo AI _CL preserves HDL function and attenuates atherosclerosis in preclinical models, supporting site-specific apo AI glycation as a mechanistically relevant feature of diabetic atherosclerosis. REGISTRATION: URL: https://www.clinicaltrials.gov ; Unique identifier: NCT05659043.

Background: Predictors for the successful eradication of neutralizing anti-FVIII alloantibodies (inhibitors) in hemophilia A patients receiving immune tolerance induction (ITI) therapy remain limited. Maternal microchimerism (MMc) showed potential to protect hemophilia A patients from inhibitor development. Objectives: To investigated the role of MMc in ITI therapy. Patients/Methods: This observational study enrolled 121 pediatric with hemophilia A and inhibitors. MMc was determined using droplet digital PCR. Low-dose ITI (FVIII ~50IU/kg every other day) was administered, with adjunctive rituximab given to patients with higher risk clinical features. Results: Of the 101 patients evaluable for MMc, 88 completed ITI, 18 were MMc positive (MMc+) and 70 were MMc negative (MMc-). Success was achieved in 75 (85.2%) patients, including 16 of 18 MMc+ patients (88.9%) and 59 of 70 MMc- patients (84.3%). Compared with MMc- patients, MMc+ patients had a lower peak inhibitor during ITI (median, 5.3 vs. 37.8 BU/ml, p = 0.029), less frequent rituximab use (27.8% vs. 61.4%; p = 0.011), and a shorter time to ITI success (median, 3.0 vs 9.9 months; p = 0.009). Multivariate Cox regression identified MMc+ (HR = 2.770), pre-ITI inhibitor F8 mutations (HR = 2.344) as independent predictors of shorter time to ITI success. Conclusion: MMc was associated with more rapid ITI success in children with hemophilia A and inhibitors receiving low-dose ITI regimen, suggesting the potential role of MMc in facilitating the eradication of FVIII inhibitors.

Background: This study aims to systematically characterize the physiological trajectories of complete blood cell count (CBC) parameters and their derived inflammatory indicators with age, providing high‐quality evidence for assessing immunosenescence and the health of the elderly. Methods: This cross‐sectional study initially recruited 9331 participants aged 24–97 years. After applying exclusion criteria, 1630 healthy individuals were included for analysis. Data are presented as median (2.5th–97.5th percentiles). The Kruskal–Wallis test followed by Dunn's test with Bonferroni correction was used to compare CBC parameters and their derived indicators among four age groups within each sex. The Mann–Whitney U test was used to compare differences between males and females within each age group. Results: Red blood cell (RBC), hemoglobin (HGB), hematocrit (HCT), and mean corpuscular hemoglobin concentration (MCHC) decreased with age, while mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), and red blood cell distribution width (RDW‐CV) increased; sex‐specific differences persisted in most parameters. Monocyte percentage and absolute count gradually increased with age; other white blood cell differentials showed variable patterns. Among CBC derived indicators: PLR and SII decreased with age; MLR remained low with limited variability in males; NLR exhibited a fluctuating trend across age groups. Platelet (PLT) progressively decreased with age. Conclusion: The comprehensive dataset generated in this study has allowed observation of dynamic profiles of CBC parameters and their derived indicators in the healthy elderly population. This provides scientific evidence and theoretical support for aging and health assessment as well as chronic disease management among older adults.

Elevated FXI levels are associated with increased risk of venous thromboembolism, yet genetic mutations contributing to elevated FXI levels have not been reported yet. We described a patient with a history of deep venous thrombosis (DVT) who tested negative on routine laboratory screening but was found to carry a novel pathogenic FXI mutation, Gly397Ser (G397S). FXI mutant protein was expressed, and functional assays were conducted in vitro to explore the underlying thrombotic mechanism. Consistent with FXI activity (FXI:C) and antigen (FXI:Ag) determined in the patient, the FXI G397S mutant exhibited a 2-fold increase of FXI:C/FXI:Ag. FXIa hydrolysis assay demonstrated that the Km value for G397S mutant was comparable to the wild-type FXIa (186.2~281.9 μM V.S. 141.6~242.6 μM), whereas the mutant FXIa displayed approximately 3-fold increases in kcat values. FXI activation by polyanions was more pronounced in the G397S mutant. Notably, this activation was more significant when triggered by thrombin compared with FXIIa in the presence of heparin. Otherwise, the G397S FXI mutant exhibited similar hydrolytic activity of FXIa against FIX, and equivalent inhibition of FXIa by Protease nexin 2 (PN2). Clot lysis assay revealed that the lysis time was longer in G397S mutant than the wild-type and the enhanced fibrinolytic resistance was thrombin-activatable fibrinolysis inhibitor-dependent. In summary, the FXI G397S mutation exhibited higher FXIa activity mainly because of the enhanced activation of FXI, which sustained thrombin generation and manifested resistance to fibrinolysis, ultimately leading to the development of DVT. ^‡ These authors contributed equally to this article. Received: 19 February 2025 Accepted after revision: 05 November 2025 Article published online: 08 December 2025 © 2025. Thieme. All rights reserved. Georg Thieme Verlag KG Oswald-Hesse-Straße 50, 70469 Stuttgart, Germany

Background: Hemophilia A (HA) arthropathy poses a significant clinical challenge, particularly in achieving perioperative hemostasis and surgical precision. Accounting for the majority of hemophilia cases, HA frequently leads to severe secondary joint damage. Although gene therapy holds promise, its clinical application in HA remains limited by challenges to durable efficacy. Crucially, the transient efficacy and unpredictable pharmacokinetics of gene therapy have not been evaluated in major orthopedic surgeries with a high potential for bleeding, such as total hip arthroplasty (THA). Case presentation: To our knowledge, this is the first documented case of robot-assisted (RA)-THA performed after gene therapy for severe HA-associated hip arthropathy, including a detailed surgical protocol and a postoperative follow-up of over six months. The patient received hemophilia gene therapy 56 weeks prior to surgery. Crucially, no exogenous FVIII supplementation was required preoperatively or intraoperatively. A minimal amount of FVIII was administered postoperatively. No abnormal bleeding events occurred throughout the perioperative period. The patient maintained normal coagulation status, achieved excellent wound healing, and demonstrated excellent functional recovery. Conclusion: This pioneering case demonstrates the potential safety and efficacy of combining gene therapy with minimally invasive RA-THA for HA hip arthropathy. The integrated approach significantly reduced dependence on exogenous FVIII and enabled precise anatomical reconstruction.

Long noncoding RNAs (lncRNAs) have been extensively studied for their regulatory roles in gene expression; however, relatively little is known about those specifically associated with super-enhancers (SEs)—genomic elements crucial for maintaining cell identity and function. This study profiles acute promyelocytic leukemia (APL)-specific SE-associated lncRNAs (SE-lncRNAs) using H3K27ac ChIP-seq and RNA-seq data from TCGA and Beat AML cohorts, identifying 44 SE-lncRNAs potentially involved in APL pathogenesis. Among these, we characterized a novel SE-lncRNA, RP11-750H9.5, located upstream of the SPI1 gene encoding the hematopoietic transcription factor PU.1, and designated it as lnc-SPI1U. Functional assays demonstrated that lnc-SPI1U suppressed differentiation, apoptosis and ATRA-induced cell proliferation inhibition, contrasting with the established role of SPI1 in promoting differentiation. Mechanistically, lnc-SPI1U interacted with heterogeneous nuclear ribonucleoproteins HNRNPH1/F, destabilizing SPI1 mRNA. Furthermore, we found that the induction of lnc-SPI1U during myeloid differentiation was dependent on PU.1, which established a feedback loop to fine-tune SPI1 expression at the optimal level for myeloid differentiation. In APL, the PML/RARα fusion protein inhibited PU.1-dependent transactivation of lnc-SPI1U by hijacking the SE region that overlapped with lnc-SPI1U locus. Our study provides mechanistic insight into how SE-lncRNAs contribute to the precise regulation of transcription factor activity during myeloid differentiation.

A 64-year-old male patient with hemophilia A was scheduled for the surgical removal of a pulmonary mass. Preoperative evaluation revealed that the coagulation factor Ⅷ (FⅧ) activity was 0.5%, with an F Ⅷ inhibitor level of 32 BU/ml; the R value could not be detected on the thromboelastogram. Thoracoscopic lobectomy was successfully completed. On the day of the operation and the first day after the operation, 6 mg of recombinant activated coagulation factor Ⅶ (rFⅦa) was intravenously administered every 6 h. On postoperative day 1, the patient's blood pressure dropped and the HGB gradually declined from 102 g/L to 65 g/L. Chest X-ray revealed a large amount of pleural effusion on the left side, and urgent thoracoscopic thoracic exploration was performed. A total of 3200 mL fresh blood was cleared, and a thoracic drainage tube was placed. On postoperative day 2, the rFⅦa dose was increased to 6 mg, which was intravenously administered every 4 h, and concentrated red cells were intermittently infused to correct anemia. Four days later, due to the inability to obtain rFⅦa, PCC (50 IU/kg every 8 hours) was administered. Additionally, treatment with methylprednisolone (40 mg/d) and cyclophosphamide (200 mg, every 2 weeks) was initiated to remove FⅧ inhibitors. The thoracic drainage tube was removed on postoperative day 9, and the patient was successfully discharged 3 weeks later.