Neutropenia is a common complication in oncology patients receiving chemotherapy, and rapid regeneration of functional neutrophils is critical for effective management. Bletilla striata polysaccharide (BSP) has shown therapeutic potential, but its mechanisms and molecular targets remain unclear. Here, we demonstrate that BSP accelerates the recovery of white blood cells, particularly neutrophils, in a chemotherapy-induced neutropenia (CIN) mouse model with cyclophosphamide (CY). The regenerated neutrophils retained phagocytic activity against bacteria, and BSP treatment significantly reduced mortality in the endotoxin-induced mouse death model. Furthermore, BSP enhanced the repopulation of hematopoietic stem and progenitor cells (HSPCs) in the bone marrow and promoted cell-cycle entry, resulting in increased frequencies of long-term hematopoietic stem cells (LT-HSCs), multipotent progenitors 2 (MPP2), and MPP3/4 subsets. Both in vitro colony formation and in vivo competitive transplantation assays confirmed that BSP reshapes hematopoietic reconstitution and corrects aberrant myeloid differentiation. PCR array analysis of HSPCs indicated that this process is mediated by C/EBPε and its downstream genes (LTF, LCN2, and ELANE). Consistently, BSP failed to support myeloid reconstitution following C/EBPε knockdown in vitro. In a C/EBPε knockout mouse model, HSPCs repopulation and regeneration were impaired, and BSP failed to promote neutrophil recovery after CY challenge or the mobilization of MPP2 and MPP3/4 subsets. The regulatory effects of BSP on C/EBPε target genes were also abolished. In conclusion, our findings identify C/EBPε as a key mediator of BSP activity, driving HSPCs repopulation and restoring hematopoietic function. These results highlight BSP as a potential therapeutic strategy for chemotherapy-induced neutropenia.

The dysregulation of intestinal lipid metabolism induced by a high-fat diet (HFD) is associated with metabolic diseases; however, the validity of intestinal organoids (IOs) as substitutes for traditional research systems remains uncertain. This study aimed to determine whether fatty acid (FA)-treated IOs accurately replicate intestinal lipid metabolism observed in HFD mice. Male C57BL/6 mice were fed either a normal chow diet (NCD) or HFD for 12 weeks, while mouse small intestinal crypt-derived IOs were treated in vitro with either an FA cocktail or a vehicle. Mice on the HFD exhibited phenotypes characteristic of metabolic syndrome, including intestinal lipid accumulation, upregulation of lipid catabolic genes, and downregulation of lipogenic genes. FA-treated IOs demonstrated enhanced budding frequency, lipid droplet accumulation, enriched lipid catabolism pathways, and suppressed lipogenesis, aligning with the in vivo findings. Omics analyses of the top 87 variable genes indicated a strong congruence between FA-treated IOs and intestinal tissues from HFD mice, with control groups clustering distinctly. The differentially expressed genes in both models were implicated in metabolic reprogramming, immune modulation, and barrier remodeling. Therefore, FA-treated IOs recapitulate key morphological and transcriptional characteristics of intestinal lipid metabolism in HFD mice, offering a valuable complementary model for investigating intestinal metabolic disorders.

Background/Objectives: Neutrophil extracellular traps (NETs) contribute to thromboinflammation and microvascular obstruction after ischemic brain injury. Borneol has anti-inflammatory and microcirculation-related pharmacological activities, but its effects on cerebral microcirculatory dysfunction and NET-related changes remain unclear. This study aimed to determine whether borneol attenuates polyethylene microsphere-induced cerebral microcirculatory dysfunction and to examine its association with NET-related markers. Methods: Rat bone marrow-derived neutrophils were stimulated with PMA with or without borneol. Male Sprague–Dawley rats were subjected to polyethylene microsphere-induced cerebral microcirculatory dysfunction and treated intragastrically with borneol (0.1, 0.2, or 0.4 g/kg). NET formation, neurological deficits, hemodynamics, cerebral microvascular perfusion, hemorheology, histopathology, inflammatory and oxidative stress markers, and NET-related molecules were assessed by immunofluorescence, ELISA, quantitative PCR, and Western blotting. Results: Borneol reduced PMA-induced NET formation in vitro. In vivo, borneol improved neurological scores, hemodynamic indices, microvascular perfusion, hemorheological parameters, and histopathological injury. It also reduced serum TNF-α, IL-1β, and ROS, decreased cerebral MPO/CitH3 signals, and downregulated MPO, PADI4/PAD4, CitH3, TNF-α, IL-6, IL-8, ICAM-1, and MIP2. Conclusions: These findings suggest that borneol alleviates polyethylene microsphere-induced cerebral microcirculatory dysfunction, accompanied by reductions in NET-related markers. However, as borneol was administered prophylactically before model induction, these results should be interpreted with caution and do not directly support a post-insult therapeutic application.

Background/Objectives: Myocardial microcirculatory dysfunction is a critical pathological feature of cardiovascular diseases, closely associated with inflammation, oxidative stress, and excessive neutrophil activation. Neutrophil extracellular traps (NETs) serve as crucial mediators of myocardial microvascular inflammatory injury. Dalbergia odorifera volatile oil (DOVO) demonstrates anti-inflammatory and antioxidant properties; however, its protective role against myocardial microcirculatory damage and its regulatory effect on NET formation remain inadequately characterized. This study investigates the protective effects of DOVO on myocardial microcirculatory disturbances and elucidates the underlying mechanisms related to NETs. Methods: A rat model of myocardial microcirculatory dysfunction was established through polyethylene microsphere injection, and an in vitro neutrophil inflammation model was generated using differentiated HL-60 cells. DOVO was administered at various doses both in vivo and in vitro, and hemodynamics, inflammatory cytokines, oxidative stress, and NET-related markers, including MPO and CitH3, were analyzed. Results: DOVO dose-dependently ameliorated microcirculatory impairment, hemodynamic disorders, inflammation, and oxidative stress in rats, significantly suppressing NET formation. In differentiated HL-60 cells, DOVO similarly reduced inflammatory gene expression and inhibited LPS-induced NETs production by downregulating MPO and CitH3. Conclusions: DOVO suggests a protective effect against myocardial microcirculatory injury by inhibiting oxidative stress, inflammatory responses, and subsequent NET formation. These findings elucidate a novel mechanism by which DOVO alleviates microcirculation-related cardiac damage and provide a theoretical basis for its application in cardiovascular injury.

Objective: This study aims to examine the association between the frailty index (FI) and stroke risk among Chinese adults, describe the burden of stroke in China, and explore the causal role of FI in ischemic stroke through Mendelian randomization. Methods: Data from the China Health and Retirement Longitudinal Study (CHARLS) included 13,473 participants aged 45 years and older without a history of stroke. Cox models, restricted cubic splines, and sensitivity analyses were employed to assess the association between the modified frailty index (mFI) and incident stroke. Additionally, data from the Global Burden of Disease (GBD) 2021 data report were utilized to describe stroke trends in China from 1990 to 2021. Two-sample Mendelian randomization was conducted to evaluate the causal effects of FI on ischemic stroke subtypes. Results: During a median follow-up period of approximately 7 years, 811 incident strokes were recorded. Each 0.1-point increase in mFI was associated with a 16% increase in stroke risk (HR = 1.16, 95% CI: 1.06–1.27), demonstrating a linear dose–response relationship. From 1990 to 2021, the proportion of ischemic stroke rose from 46.9% to 63.2%. Mendelian randomization analysis provided genetic evidence supporting a causal association between FI and ischemic stroke (OR = 1.191, 95% CI: 1.046–1.357), particularly driven by large-artery atherosclerotic (OR = 1.852) and small-vessel stroke (OR = 1.415), but not by cardioembolic stroke. Conclusions: A higher FI is associated with an increased risk of stroke among Chinese adults, with genetic evidence supporting a causal role in ischemic stroke. Therefore, FI may serve as a valuable addition to existing risk assessment tools.