Retinitis pigmentosa (RP) exhibits complex genetic heterogeneity, and the downstream pathogenic mechanisms underlying different etiologies remain unclear. This study aimed to explore the differential mechanisms by which the metabolite uridine and its receptor P2Y6R regulate microglial activation and photoreceptor apoptosis in different RP animal models.

① Uridine content in the retina was measured using ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) in rd1 mice (aged 28 d, weighing 13 to 18 g, n=3), rd10 mice (aged 45 d, weighing 20 to 25 g, n=3) and age-matched C57BL/6J control mice (n=3/group), as well as RCS rats (aged 60 d, weighing 200 to 250 g, n=4) and age-matched RCS-rdy control rats (n=4). Retinal pyrimidine metabolites were analyzed using ultra performance liquid chromatography-high resolution mass spectrometry (UPLC-HRMS) at various disease stages in rd10 mice (aged 18 d, weighing 7 to 10 g; aged 25 d, weighing 11 to 15 g; aged 45 d; n=5), with age-matched C57BL/6J mice serving as controls (n=5). ② Five-week-old C57BL/6J mice received unilateral subretinal injection of uridine (100 mmol/L, 2 μL), with contralateral eyes injected with PBS as controls. After 7 d, electroretinography (ERG), outer nuclear layer (ONL) thickness analysis, immunofluorescence staining, and TUNEL assay were performed (n=3). ③ RT-qPCR was used to detect mRNA expression of P2Y receptor family members (P2ry1, P2ry2, P2ry4, P2ry6, P2ry12) in the retina of C57BL/6J mice following subretinal injection of uridine or PBS (n=3). ④ The P2Y6R inhibitor MRS2578 (3 mg/kg per day) or vehicle (corn oil containing 0.6% dimethyl sulfoxide) was administered to rd10 mice (aged 14 d, weighing 7 to 10 g, n=3) and RCS rats (aged 15 d, weighing 25 to 30 g, n=3) prior to degeneration onset. ERG, ONL thickness, photoreceptor apoptosis, and microglial number and morphology were evaluated. ⑤ RT-qPCR was performed to detect the expression of uridine synthesis and metabolism-related enzymes (Upp1, Upp2, Uck1, Uck2) in the retina of 45-day-old rd10 and C57BL/6J mice (n=3).

① Compared with age-matched C57BL/6J control mice, retinal uridine content was significantly elevated in rd1 and rd10 mice (P<0.05), whereas no significant change was observed in RCS rats compared with age-matched RCS-rdy control rats. Retinal uridine levels in rd10 mice progressively increased with disease progression compared with age-matched C57BL/6J control mice (P<0.000 1). ② Subretinal injection of uridine in C57BL/6J mice resulted in significant reductions in a-wave and b-wave amplitudes (P<0.05), ONL thinning (P<0.000 1), increased microglial activation (P<0.01) with migration toward the ONL, and increased TUNEL-positive cells (P<0.01). ③ After uridine injection, the mRNA expression of P2ry6 was upregulated in the retina of C57BL/6J mice (P<0.05). ④ Compared with the vehicle-treated group, MRS2578 treatment significantly improved the amplitudes of the a-wave and b-wave (P<0.05), increased the ONL thickness, reduced the number of TUNEL-positive cells (P<0.05), and promoted the reversal of microglia from an amoeboid-activated state to a resting, ramified state in rd10 mice. In contrast, in RCS rats, MRS2578 treatment failed to significantly inhibit microglial activation or delay retinal degeneration compared with vehicle-treated controls. ⑤ Compared with age-matched C57BL/6J control mice, rd10 mice exhibited upregulated expression of Upp1 (P<0.05), Upp2 (P<0.01), and Uck1 (P<0.01) in the retina, with decreased UMP and uracil levels.

The uridine-P2Y6R signaling axis exacerbates retinal degeneration through microglial activation in the rd10 mouse model of primary photoreceptor degeneration, whereas this mechanism is absent in the RCS rat model of primary retinal pigment epithelial dysfunction. These findings reveal the heterogeneity of retinal degeneration mechanisms across different etiological backgrounds, suggesting that targeting the uridine-P2Y6R axis may represent a potential therapeutic strategy for specific RP subtypes.

Blast-induced traumatic brain injury (bTBI) is frequently observed in industrial accidents and military operations, and presents unique mechanisms of injury and complex pathological evolution distinct from conventional ocular trauma when complicated by visual dysfunction. By investigating the effects of NLRP3 inhibitor MCC950 on retinal structure and function following blast wave exposure.

A bTBI mouse model was established using a biological shock tube. Male C57BL/6J mice (8 weeks old, weighing 23.5±1.5 g) were stratified by body mass and randomly divided into a sham group, a bTBI 28 d group, a bTBI 28 d+DMSO group, and a bTBI 28 d + MCC950 group (MCC950, 10 mg/kg, i.p.), with 8 animals in each group. After blast exposure, retinal function was evaluated by electroretinography (ERG, n=8); the protein levels of NLRP3, Caspase-1, Cleaved-GSDMD, IL-1β and IL-18 in the retina were detected by Western blotting (n=3); the activation of Müller cells and microglia was analyzed by immunofluorescence staining (IF) and Western blotting (n=3); and photoreceptor ultrastructural changes were observed with transmission electron microscopy (TEM, n=3).

Compared with the sham group, the bTBI 28 d group exhibited progressive visual dysfunction, manifested by significantly reduced amplitudes of the a-wave (P<0.001), b-wave (P<0.01), and oscillatory potentials (OPs) wave (P<0.001) in dark-adapted ERG; significant activation of Müller cells (P<0.05) and microglia (P<0.001) in the retina; and significant elevated retinal protein levels of NLRP3 (P<0.05), Caspase-1 (P<0.01), Cleaved-GSDMD (P<0.05), and downstream inflammatory factors IL-1β (P<0.01) and IL-18 (P<0.001). TEM revealed disorganized outer segment disc structures and inner segment vacuolization in photoreceptors of the bTBI 28 d group. MCC950 intervention significantly reversed these changes, as evidenced by significantly increased amplitudes of the a-wave, b-wave, and OPs wave in ERG in the bTBI 28 d+MCC950 group compared with the bTBI 28 d group (P<0.05), significantly inhibited activation of Müller cells (P<0.01) and microglia (P<0.05), significant downregulation of pyroptosis pathway proteins including NLRP3, Caspase-1, Cleaved-GSDMD, IL-1β, and IL-18 (P<0.01), and markedly attenuated photoreceptor ultrastructural damage.

Retinal injury following blast exposure is closely associated with NLRP3-Caspase-1-GSDMD signaling axis-mediated photoreceptor pyroptosis. Early administration of MCC950 to inhibit this pyroptosis pathway can effectively alleviate neuroinflammation and gliosis, thereby exerting protective effects on the structure and function of retinal photoreceptor cells.

Acute graft-versus-host disease (aGVHD) is a common immunological complication following allogeneic hematopoietic stem cell transplantation (allo-HSCT). This study aims to investigate the intervention effects of 6-gingerol on aGVHD and explore its potential mechanism in regulating T cell function and differentiation.

A murine allo-HSCT aGVHD model was established. SPF-grade male mice (8 to 12 weeks old, weighing 22 to 24 g) were randomly divided into a syngeneic control group (n=5), a vehicle control group (n=8,13), and 6-gingerol experimental groups at different concentrations (n=8,13). The survival status and body weight changes of recipient mice were observed, clinical scores and target organ histopathological scores were performed, and flow cytometry was used to detect changes in T cell subsets and surface markers in the spleen and liver. A murine B cell lymphoma cell line A20 graft-versus-leukemia (GVL) model was established, and tumor growth was evaluated through in vivo imaging and survival analysis. Splenic T cells were isolated and cultured in vitro, and divided into unstimulated, stimulated, and different concentrations of 6-gingerol treatment groups to assess proliferation, apoptosis, and cytokine changes. Mouse splenic CD4⁺ T cells were isolated and cultured under induction conditions for T helper 1 (Th1) cells, T helper 17 (Th17) cells, and T regulatory (Treg) cells, and were divided into a stimulated group and experimental groups with different concentrations of 6-gingerol. Changes in the differentiation direction of CD4⁺ T cells were analyzed via flow cytometry. Bulk RNA sequencing (bulk RNA-seq) was used to analyze gene expression profiles and differential pathway changes, and the transcriptomic results were validated using RT-qPCR. Receptor agonist intervention was further performed, and then the experiment was divided into: a stimulation group, a 6-gingerol monotherapy group, a 2-MeS-ADP monotherapy group, and a 6-gingerol + 2-MeS-ADP dual-drug group. The related mechanisms were validated through T-cell proliferation assays, in vitro induced differentiation assays, detection of cytoplasmic and mitochondrial Ca²⁺, and transmission electron microscopy (TEM) and quantitative analysis of mitochondrial cristae structure.

Following 6-gingerol intervention, the survival status, body weight, and clinical scores of aGVHD mice were significantly improved in the aGVHD mice (P<0.05). HE stained pathological section scores revealed alleviated liver and intestinal injury (P<0.05). Survival analysis results and in vivo bioluminescence imaging in the GVL model showed that 6-gingerol had no significant effect on the GVL effect. In vitro experiments demonstrated that 6-gingerol suppressed T cell proliferation (P<0.000 1) and promoted apoptosis (P<0.01) in partially activated T cells. Under specific polarization conditions, 6-gingerol regulated CD4⁺ T cell subset differentiation, manifested as inhibition of Th1 (P<0.05) and Th17 (P<0.05) differentiation and promotion of Treg cell differentiation (P<0.05). Transcriptomic analysis suggested that 6-gingerol treatment affected purinergic receptor and Ca²⁺-related pathways. RT-qPCR results showed that P2ry13 expression was significantly downregulated (P<0.000 1). Further mechanistic studies revealed that following 6-gingerol treatment, cytoplasmic Ca²⁺ levels in CD4⁺ T cells decreased (P=0.000 1), while mitochondrial Ca²⁺ accumulation in CD4⁺ T cells increased (P=0.000 2). Compared with the 6-gingerol monotherapy group, the two-drug treatment group, with the addition of the P2Y13 agonist 2-MeS-ADP showed partial recovery in T cell proliferation (P<0.000 1), Th1 and Th17/Treg differentiation (P<0.05), and cytoplasmic and mitochondrial Ca²⁺ concentrations (P<0.05). TEM revealed that mitochondrial ultrastructure in the two-drug group was improved compared to that of the 6-gingerol group, and quantitative analysis of mitochondrial cristae also supported this change (P<0.000 1), suggesting that 6-gingerol regulates T cell subset differentiation through the P2Y13-related Ca²⁺ signaling pathway.

6-Gingerol mitigates aGVHD by inhibiting Th17 cells and expanding Treg cells through the purinergic receptor P2Y13-related Ca²⁺ signaling pathway, providing a novel strategy for the prevention and treatment of aGVHD.

Sialylation, an important post-translational modification, plays a key role in processes such as cell recognition and signal transduction. This study aims to explore the role of α-2,3 sialylation in osteogenic and chondrogenic differentiation, further elucidate its impact on bone development by regulating extracellular matrix (ECM) homeostasis, and provide novel theoretical basis and potential intervention targets for disorders of skeletal development.

The expression profiles of sialyltransferase family genes in osteogenic and chondrogenic differentiation were analyzed using public datasets GSE109503 (chondrogenic differentiation) and GSE113253 (osteogenic differentiation). In vitro differentiation experiments were conducted using MC3T3-E1 osteoblast and ATDC5 cells, and qRT-PCR was performed to screen for core differentially expressed genes (DEGs) in the sialyltransferase family (screening threshold |Log₂FC|>1, P<0.05). Stable α-2,3-type sialylation-deficient MC3T3-E1 and ATDC5 cells were constructed using lentiviral vectors, followed by osteogenic or chondrogenic induction for 7 and 14 d. Differentiation capacity was evaluated by ALP staining, Alizarin Red S staining, and Alcian blue staining. The expression of osteogenic marker genes (Col1a1, Sp7) and chondrogenic marker gene (Col2a1) was detected by qRT-PCR and Western blotting. Transcriptome sequencing was performed on α-2,3 sialylation-deficient MC3T3-E1 cells (osteogenic induction for 7 d) and ATDC5 cells (chondrogenic induction for 7 d), and biological functions of DEGs were analyzed with GO and KEGG enrichment analyses. For in vivo experiments, newborn C57BL/6 mice (half male and half female, body weight 1.0 to 1.5 g, on the day of birth) were randomly divided into 2 groups: an sialidase-treated group (intraperitoneally injected with sialidase, 0.2 U/dose, every other day for 4 weeks), and a control group (same volume of normal saline), with 6 animals in each group. Micro-CT scanning was conducted to analyze the bone microstructure parameters of the femur.

Public dataset analysis revealed that only St3gal1 was significantly upregulated during both osteogenic and chondrogenic differentiation (|Log₂FC| >1, P<0.05). The in vitro cell differentiation model confirmed that St3gal1was significantly upregulated in both differentiation processes (P<0.000 1). Consistent with the public dataset transcriptome sequencing results, this demonstrated that St3gal1-mediated α-2,3-type sialylation accompanies the progression of osteogenic and chondrogenic differentiation. Following disruption of α-2,3 sialylation, ALP staining and Alizarin Red S staining in MC3T3-E1 cells were significantly attenuated, along with the mRNA expression levels of Col1a1and Sp7reduced to 60.7% (P<0.000 1) and 49% (P<0.000 1) of the control group, respectively, and corresponding protein levels decreasing to 61.5% (P=0.000 2) and 56.1% (P=0.000 3), respectively. In ATDC5 cells, Alcian Blue staining was markedly diminished, and CollagenⅡ protein expression was reduced to 43.1% of the control group (P<0.000 1). Transcriptomic sequencing results demonstrated that DEGs in both cell types were significantly enriched to ECM structure, and molecular functions were associated with ECM structural constituent. Pathway enrichment analyses revealed that DEGs were significantly enriched in ECM-receptor interaction and related signaling pathways (|Log₂FC|>1, FDR<0.05), suggesting that sialylation may regulate osteogenic and chondrogenic differentiation through multiple pathways, including ECM homeostasis, thereby contributing to bone development. In vivo experiments confirmed that, the sialidase-treated group exhibited reductions in femoral bone mineral density (decreased by 0.078 g/cm³, P<0.000 1), trabecular bone volume fraction (decreased by 5.512%, P<0.000 1), cortical thickness (decreased by 0.012 mm, P=0.001 1), trabecular number (decreased by 1.39/mm, P<0.000 1), and trabecular thickness (decreased by 0.002 3 mm, P=0.026 7), and an increase in trabecular separation (increased by 0.35 mm, P<0.000 1) when compared with the control group, indicating impaired skeletal development.

α-2,3 Sialylation positively regulates osteogenic and chondrogenic differentiation through modulating ECM homeostasis and plays an essential role in skeletal development. Sialylation within the bone microenvironment may serve as a potential intervention target for skeletal dysplasia disorders.

Intervertebral disc degeneration (IVDD) is a primary cause of low back pain, and stemness exhaustion of nucleus pulposus stem cells (NPSCs) represents a key underlying mechanism. Thus, maintaining the quiescent state of NPSCs is of great significance in delaying disc degeneration. This study aimed to investigate the inductive effect of in vitro three‑dimensional (3D) spheroid culture on the quiescent state of NPSCs and its regulatory mechanism, thereby providing experimental evidence for targeted intervention in IVDD.

Intervertebral disc tissues were collected from juvenile and adult patients undergoing lumbar surgery in our department between December 2025 and January 2026. Tissue sections were prepared, and morphological differences were observed through HE staining. NPSCs were isolated from 24 male SD rats (8 weeks old, weighing 350±50 g). Stem cell immunophenotypes were identified by flow cytometry, and the stemness of NPSCs was verified by trilineage differentiation assay. NPSCs cultured under routine 2D conditions served as the monolayer culture (ML) group, while NPSCs cultured using the 3D spheroid culture system were established as the spheroid culture (SP) group. Cell morphology was observed under a microscope, cell cycle distribution was analyzed by flow cytometry, and the quiescent state was identified by EdU staining. Bulk transcriptome sequencing and bioinformatic analysis were performed on NPSCs from the ML and SP groups. The expression of autophagy‑related molecules and key molecules in the PI3K‑Akt‑mTOR signaling pathway, as well as the ultrastructure of autolysosomes, were detected by RT‑qPCR, Western blotting and transmission electron microscopy (TEM), respectively.

Clinical histological observation showed that nucleus pulposus cells in juvenile patients were scattered in distribution, whereas those in adult patients exhibited clustered distribution, and the proportion of clustered nucleus pulposus cells was significantly higher in adult patients than in juvenile patients(P<0.01). The successfully isolated rat NPSCs highly expressed mesenchymal stem cell‑positive markers CD90 (99.4%), CD44 (99.8%) and CD73 (94.2%), and lowly expressed hematopoietic stem cell‑negative markers CD34 (0.21%), CD45 (0.45%) and CD11 (1.08%) as determined by flow cytometry. Trilineage differentiation assays confirmed that NPSCs possessed osteogenic, chondrogenic and adipogenic differentiation potentials, consistent with the phenotypic characteristics of mesenchymal stem cells. In vitro spheroid culture induced NPSCs to form regular spheroids with smooth edges, with an average diameter of 52.3±11.6 μm. Cell cycle analysis revealed that the proportion of G0‑phase cells in the SP group was significantly higher than that in the ML group (P<0.05). Moreover, EdU‑positive cell proportion was markedly lower in the SP group than in the ML group (P<0.05), indicating a typical quiescent state. RT-qPCR showed that the expression of P27, SOX2 and CD34 in the SP group were significantly higher than those in the ML group (P<0.05). Bulk RNA‑sequencing demonstrated significant differentially expressed genes between the SP and ML groups, with significant enrichment of the autophagy pathway and the PI3K‑Akt‑mTOR pathway. Western blotting demonstrated that, compared with the ML group, the ratio of LC3‑Ⅱ/LC3‑Ⅰ was significantly increased, while the expression of p62 was notably decreased in the SP group. The expression levels of key molecules in the PI3K-Akt-mTOR pathway, such as PI3K, p‑Akt and p‑mTOR were all significantly downregulated (P<0.05). TEM revealed a markedly higher number of autolysosomes in the SP group than in the ML group. Rescue experiments further verified the regulatory relationship between the PI3K‑Akt‑mTOR pathway and autophagy (P<0.05). Clinical sample detection showed that the expression levels of Ki67 and p‑mTOR in nucleus pulposus tissues were significantly higher in juvenile patients than in adult patients, whereas the content of LC3B was significantly lower (P<0.05).

In vitro 3D spheroid culture can effectively induce NPSCs to enter a quiescent state and maintain their stemness. The underlying mechanism may involve the activation of autophagy via inhibition of the PI3K‑Akt‑mTOR pathway.

Currently, endovascular interventional therapy represents the primary clinical approach for vascular lesions. Restenosis caused by mechanical procedures, however, has become increasingly prominent. This study aimed to explore the feasibility and underlying mechanisms of naked plasmid encoding hepatocyte growth factor (HGF) for preventing post-interventional luminal restenosis through a rat animal model and in vitro experiments.

① A rat carotid artery injury model was established by pulling inflated balloon. Eighteen male SD rats (6 to 8 weeks old, weighing 260 to 280 g) were randomly divided into a sham operation group, normal saline group, and HGF group (adentitial injection of HGF naked plasmid) (n=6 for each group). Peak systolic velocity was measured using carotid ultrasound, and the area of the vascular intima and HGF expression were assessed through HE staining and HGF immunofluorescence staining, respectively. The expression levels of phenotypic markers in the carotid media were determined by Western blotting. ② Vascular endothelial cells (ECs) and vascular smooth muscle cells (VSMCs) were obtained from rat carotid arteries through primary culture, and both ECs and VSMCs were transfected with the plasmids haboring HGF. The proliferative and migratory abilities of transfected and untransfected ECs were assessed through CCK-8 assay and cell scratch assay, respectively. After Ang Ⅱ-induced VSMC transformation, the effects of Ang Ⅱ induction alone, Ang Ⅱ induction followed by HGF plasmid transfection, Ang Ⅱ induction followed by exogenous HGF treatment on VSMC migratory ability were detected by cell scratch assay, and the expression levels of phenotypic markers in VSMCs were determined by RT-qPCR and Western blotting.

① Animal experiments showed that, compared with the normal saline group, the peak systolic velocity of the carotid artery in the HGF group was significantly lower on post-injury day 3 and day 7 (day 3: 422.62±16.99 vs 1 050.58±40.92 mm/s; day 7: 631.81±26.11 vs 1 309.78±73.13 mm/s; P<0.001), the increase in carotid artery intimal area was significantly less (day 3: 0.16±0.03 vs 0.83±0.09 mm²; day 7: 0.17±0.03 vs 1.15±0.31 mm²; P<0.001), the green fluorescence intensity of HGF in the carotid artery tissue was significantly higher (P<0.001), and the protein expression levels of synthetic phenotype markers vimentin, OPN, and PCNA were all significantly lower (P<0.001), whereas the protein expression level of contractile phenotype marker α-SMA was comparable (P>0.05). ② Cell experiment results showed that, compared with untransfected ECs, HGF plasmid-transfected ECs exhibited significantly enhanced proliferative activity and migratory ability (P<0.05). Compared with VSMCs receiving Ang Ⅱ induction alone, both Ang Ⅱ-induced VSMCs subsequently transfected with HGF plasmid and those treated with exogenous HGF demonstrated significantly reduced migratory ability (P<0.01), significantly decreased mRNA and protein expression levels of the synthetic phenotype marker vimentin (P<0.05), and significantly elevated mRNA and protein expression levels of the contractile phenotype marker SM-MHC (P<0.05).

HGF may promote vascular re-endothelialization and inhibit intimal hyperplasia by enhancing the proliferative and migratory abilities of vascular ECs and inhibiting the transition of VSMCs to a synthetic phenotype.

Colorectal cancer (CRC) is a cold tumor with a low response rate to programmed death-ligand 1 (PD-L1) inhibitors, and the regulatory mechanism underlying its expression remains incompletely understood. Lactylation is a novel post-translational modification involved in the regulation of gene transcription and protein function. This study aims to investigate the correlation between lactylation levels and PD-L1 expression in CRC, as well as its clinicopathological significance, so as to provide a novel potential target for immunotherapy.

① The GEPIA2 database was used to analyze the expression correlation between CD274 (encoding PD-L1) and 18 glycolysis genes (including HK1, HK2, and 16 others), lactate dehydrogenase genes (LDHA, LDHB), and lactate transporter genes (SLC16A1, SLC16A4). The TCGA colon adenocarcinoma (COAD) dataset with 453 samples was obtained from the XENA website, and GSEA software was applied to analyze gene ontology_biological process (GO_BP) pathway differences between the CD274 high- and low-expression groups. The Kaplan-Meier plotter database was used to evaluate the correlation between CD274 expression and overall survival in 4 consensus molecular subtypes (CMS) of CRC. ② Human CRC cell lines HT29 and SW480 were treated with 10 mmol/L lactic acid (Lac) or 10 mmol/L sodium lactate (Nala) for 24 h, with corresponding blank control groups, or cultured in low-glucose medium (1.8 g/L glucose) or high-glucose medium (4.5 g/L glucose). The transcription and translation levels of PD-L1 were detected by RT-qPCR and Western blotting, respectively. In addition, cells were treated with gradient concentrations of sodium lactate or Oxamate (a lactate dehydrogenase inhibitor) to verify the dose-dependent regulation of lactylation on PD-L1 expression. ③ Cancer tissues and paired adjacent non-tumor mucosal tissues were collected from 35 CRC patients (aged 59±11 years, a male-to-female ratio of 2.2∶1.0) who underwent surgical treatment at Department of General Surgery of First Affiliated Hospital of Army Medical University between February 2024 and October 2025. Immunohistochemistry (IHC) was performed to detect pan-lactylation (Pan-Kla) and PD-L1 expression, expression levels were evaluated by IHC scoring, and their correlations were analyzed with clinicopathological features.

① GEPIA2 database analysis demonstrated that CD274 expression was significantly positively correlated with glycolysis and lactate dehydrogenase genes (R=0.14 to 0.47, P<0.05), including hexokinase 1 (HK1), hexokinase 3 (HK3), phosphofructokinase platelet type (PFKP), triosephosphate isomerase 1 (TPI1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), phosphoglucomutase 2 (PGM2), phosphoglucomutase 3 (PGM3), enolase 1 (ENO1), enolase 2 (ENO2), pyruvate kinase M (PKM), lactate dehydrogenase A (LDHA), and lactate dehydrogenase B (LDHB). GSEA revealed that the glycolysis pathway was significantly activated in the CD274 high-expression group, while the oxidative phosphorylation pathway showed no obvious changes. Kaplan-Meier analysis indicated that in CMS Ⅰ (microsatellite unstable) and CMS Ⅳ (mesenchymal) subtypes, the CD274 low-expression group had significantly longer overall survival, whereas no significant correlation was found in CMS Ⅱ (canonical) and CMS Ⅲ (metabolic) subtypes. ② After treatment with Lac and Nala, the mRNA and protein levels of PD-L1 were significantly increased in HT29 and SW480 cells compared with the control group. Cells cultured in low-glucose medium exhibited lower pan-lactylation and PD-L1 expression levels than those cultured in high-glucose medium. Sodium lactate upregulated PD-L1 expression in a dose-dependent manner, and Oxamate treatment reduced PD-L1 expression. ③ Pan-lactylation and PD-L1 scores were significantly higher in CRC tissues than in paired adjacent non-tumor tissues (P<0.01). A significant positive correlation was observed between them (R²=0.195 9, P=0.007 8). Clinicopathological correlation analysis showed that mucinous adenocarcinoma had significantly higher lactylation scores than non-mucinous adenocarcinoma (P=0.047).

Lactylation upregulates the transcriptional and translational levels of PD-L1 in colorectal cancer cells and is significantly positively correlated with PD-L1 expression in tumor tissues. The prognostic value of PD-L1 is CMS subtype-specific, and low PD-L1 expression indicates a better prognosis in CMS Ⅰ/Ⅳ patients.

Brucella abortus (B.abortus) is one of the primary causative agents of brucellosis, posing potential threats to livestock production and public health security. This study aims to screen specific M13 phages targeting the bacteria and to establish a phage-functionalized electrochemical biosensor, thereby validating its detection performance and application potential in actual sample testing.

Using B.abortus as a target, biopanning of a phage display peptide library was performed, combined with negative screening to enrich candidate M13 phage clones. Indirect ELISA was employed to assess the affinity and specificity of each clone. The phages were immobilized onto a gold electrode surface, with bovine serum albumin used to block non-specific sites to construct an electrochemical biosensor. Differential pulse voltammetry (DPV) current response values were employed as an indicator to optimize phage titer, phage incubation time, and sample incubation time. The DPV current response values were measured for concentrations ranging from 1.0×10³ to 1.0×10⁷ CFU/mL. A four-parameter logistic function was used to fit the standard curve, and the limit of detection (LOD) was calculated. Staphylococcus aureus, Staphylococcus epidermidis, Acinetobacter baumannii, and Escherichia coli were selected as negative controls to evaluate the specificity of the sensor. The same concentration of B.abortus samples was repeatedly detected 10 times, and the relative standard deviation (RSD) was calculated to assess repeatability. The sensor was stored at 4 ℃, and B.abortus was detected at different time points to calculate the RSD and evaluate the stability. B.abortus bacterial suspensions were prepared as spiked serum samples using fetal bovine serum as the matrix. SPF female BALB/c mice (6 to 8 weeks old, weighing 16 to 18 g) were randomly divided into a control group and an infected group, with 5 animals in each group. The infection group was intraperitoneally inoculated with B.abortus, and the blank group was inoculated with the same volume of PBS. Mouse spleen samples were collected. Both the constructed electrochemical biosensor and plate counting method were used to detect the above 2 types of samples. The recovery rate was calculated to evaluate the detection accuracy of the sensor in actual samples.

Through biopanning and indirect ELISA validation, M13 phage clone P-1 capable of specifically binding to B. abortus was screened, and its binding affinity to B. abortus was superior to that of other control strains. The optimal construction conditions for the sensor were as follows: phage titer of 1.0×10¹¹ PFU/mL, phage incubation time of 2 h, and sample incubation time of 1 h. The fitting equation for this sensor within the bacterial concentration range of 1.0×10³ to 1.0×10⁷ CFU/mL is \mathrm{I}/\mathrm{\mu }\mathrm{A}=\mathrm{81.13}+\frac{(\mathrm{184.08}-\mathrm{81.13})}{{\left(\mathrm{1}+\frac{\mathrm{L}\mathrm{o}{\mathrm{g}}_{\mathrm{10}}\mathrm{C}}{\mathrm{5.01}}\right)}^{\mathrm{8.68}}} (R²= 0.999 9), with an LOD of 157 CFU/mL. The sensor demonstrated favorable detection specificity (P<0.000 1) and repeatability (RSD=2.70%), with stable performance (RSD=2.37%). The recovery rate of B.abortus detected by the sensor in fetal bovine serum samples ranged from 98.2% to 106.0%, while the bacterial load in spleen samples from infected mouse model was detected as 1.23×10³ CFU/mL, and the accuracy of this method was consistent with the gold standard plate culture method.

A specific M13 phage clone, P-1, targeting B. abortus was identified through screening, and this clone was used to construct an electrochemical biosensor capable of quantitatively detecting the target bacterium.

Hepatopulmonary syndrome (HPS) is characterized by inflammation, pulmonary vasodiation and angiogenesis. Elevated serum level of fatty acid binding protein 4 (FABP4) in patients with liver cirrhosis is associated with poor prognosis, and FABP4 is involved in the regulation of inflammation and angiogenesis. Using a bile duct ligation (BDL)-induced HPS rat model and the FABP4 inhibitor BMS309403 for intervention, this study aims to investigate the role of the FABP4/PPAR-γ signaling axis in the pathogenesis of HPS and to evaluate the protective mechanism of FABP4 inhibition against hepatopulmonary injury.

A total of 24 male SPF SD rats (6 to 8 weeks old, weighing 200 to 220 g) were enrolled and randomly divided into (n=6): a sham operation group (Sham group), the BDL postoperative 3-week group (BDL3W group), the BDL postoperative 5-week group (BDL5W group), and a BDL+BMS309403 treatment group (BDLB group). The BDL3W, BDL5W and BDLB groups underwent BDL to establish an HPS model, while the Sham group underwent only bile duct manipulation without ligation. The BDLB group received intraperitoneal injections of BMS309403 (5 mg/kg, twice per week) from postoperative day 15 for 3 weeks. The BDL3W group was sacrificed at week 3 post-surgery, while the Sham, BDL5W, and BDLB groups were sacrificed at week 5 post-surgery. Histopathological changes in the liver and lung tissues were evaluated using HE staining. The severity of hepatic fibrosis was assessed by Masson’s trichrome staining, while hepatic lipid accumulation was examined using Oil Red O staining. Microarray was performed on lung tissues from the Sham and BDL3W groups, and integrated analysis of differentially expressed genes (DEGs) was conducted, including: heatmap plotting based on specific gene sets, visualization of differential expression with volcano plots, and GO functional enrichment analysis. Immunohistochemistry was used to detect the localization and expression levels of CD31, iNOS, TNF-α, NF-κB p65, and PPAR-γ in lung tissues. Immunofluorescence staining was performed to determine the expression of FABP4. The protein levels of FABP4 and PPAR-γ in lung tissues were quantified by Western blotting. Arterial blood gas analysis was conducted to measure PaO₂ and P(A-a)O₂ for the evaluation of pulmonary function, and serum levels of ALT and AST were determined to evaluate hepatic function.

① Compared with the BDL3W group, the BDL5W group exhibited progressively disorganized liver structure, gradually aggravated fibrosis, and a significant increase in relative collagen areas (P<0.01). Serum ALT levels increased continuously from the Sham group to the BDL3W and BDL5W groups (P<0.01), with AST levels showing the same trend (P<0.01). In the lungs, inflammatory cell infiltration aggravated progressively, and PaO₂ decreased gradually from the Sham to BDL3W and BDL5W groups (P<0.05). Compared with the BDL3W group, P(A-a)O₂ was markedly elevated in the BDL5W group (P<0.01), confirming the successful establishment of the HPS model. ② In the BDL3W group, FABP4 gene was significantly upregulated in the lung tissues (FC≥1.5, P≤0.05). GO enrichment analysis revealed significant enrichment of lipid metabolism-related genes, suggesting that FABP4 and lipid metabolic processes play important roles in the pulmonary pathogenesis during HPS. ③Compared with the BDL5W group, the BDLB group showed significantly decreased serum AST and ALT levels (P<0.01), accompanied by attenuated hepatic fibrosis, as evidenced by a significant reduction in relative collagen area (P<0.01).Pulmonary inflammatory infiltration, hemorrhage, and atelectasis were alleviated, with a significant increase in PaO₂ (P<0.01) and an obvious decrease in P(A-a)O₂ (P<0.01). ④ Immunofluorescence and Western blotting revealed that FABP4 expression in the lung tissues of the BDLB group was significantly lower than that in the BDL5W group (P<0.05). Immunohistochemistry and Western blotting further demonstrated that PPAR-γ expression in the lung tissues was significantly enhanced in the BDLB group compared with the BDL5W group (P<0.05). ⑤ Compared with the BDL5W group, the BDLB group exhibited reduced hepatic lipid droplet accumulation. Immunohistochemical analysis showed a significant decrease in the diameter of pulmonary microvascular lumens (P<0.01), along with significantly downregulated expression of iNOS (P<0.01), and remarkably reduced expression levels of the inflammatory factors TNF-α and NF-κB p65 (P<0.01).

The FABP4 inhibitor BMS309403 ameliorates hepatopulmonary pathological injury and improves hypoxemia in HPS rats by modulating the FABP4/PPAR-γ signaling axis, ameliorating hepatic lipid metabolism disorder, inhibiting NF-κB p65/TNF-α-mediated pulmonary inflammatory response, and suppressing iNOS/NO-driven pulmonary microvascular dilation.

Cognitive impairment is common after intracerebral hemorrhage (ICH), yet conventional imaging metrics fail to explain its neural mechanisms from the perspective of whole-brain functional networks. This study aims to investigate abnormal changes in small-world topological properties of resting-state functional brain networks in patients with spontaneous ICH during the subacute phase, to characterize the topological reorganization at both global and nodal levels, and to further elucidate the relationship between differential topological metrics and cognitive impairment as well as the influence of lesion volume.

An observational cross-sectional study was conducted. A total of 28 patients with subacute spontaneous ICH (ICH group) who were treated at Department of Neurosurgery of Huizhou Central People’s Hospital between December 2024 and January 2026, and 35 healthy controls (HC) matched for age, sex, and years of education who were treated at Health Check-up Center or General Clinic of Huizhou Central People’s Hospital during the same period were enrolled. All participants underwent Mini-Mental State Examination (MMSE) assessment and 3.0 T resting-state functional MRI (rs-fMRI) scanning. For ICH patients, T1 structural images were subjected to semi-automatic lesion delineation using ITK-SNAP software, and mirror normalization was performed based on the Clinical Toolbox for SPM to reduce interference from focal lesions on spatial normalization. rs-fMRI data were preprocessed using the DPABI software package, including head motion correction, tissue segmentation, spatial normalization, linear detrending, nuisance regression, and bandpass filtering. When extracting the mean BOLD time series from 116 brain regions of interest defined by the automated anatomical labeling (AAL) atlas, voxels falling within the lesion and hematoma masks were excluded. Functional connectivity matrices were constructed using Pearson correlation coefficients followed by Fisher r-to-z transformation. The GRETNA toolbox was used to construct binary undirected functional brain networks within a sparsity threshold range of 0.05 to 0.40, and the area under the curve (AUC) was calculated for global topological metrics including clustering coefficient (Cp), characteristic path length (Lp), normalized clustering coefficient (Gamma), normalized characteristic path length (Lambda), small-world property (Sigma), global efficiency (Eglob), and local efficiency (Eloc), as well as nodal metrics including nodal efficiency (Ne) and nodal local efficiency (NLe).

Both groups exhibited small-world topological characteristics (Lambda≈1, Sigma>1); however, the ICH group demonstrated significant decreases in Sigma, Gamma, Eloc, Eglob, and Lambda (all FDR-corrected P<0.05), an increase in Lp (FDR-corrected P=0.001), and no statistical difference in Cp (FDR-corrected P=0.086). At the nodal level, following Bonferroni correction, the ICH group exhibited decreased nodal efficiency in the left globus pallidus, bilateral amygdala, right posterior cingulate gyrus, left putamen, left cerebellar CrusⅠ, and cerebellar vermis (P<0.05), and increased nodal efficiency in the left medial superior frontal gyrus and left supplementary motor area (P<0.05). Regarding nodal local efficiency, the left globus pallidus, left amygdala, and right posterior cingulate gyrus were decreased (P<0.05). Correlation analysis revealed that nodal efficiency of the left medial superior frontal gyrus was negatively correlated with spatial orientation scores (r=-0.452, P=0.023), nodal efficiency of the left amygdala was negatively correlated with tongue twister repetition scores (r=-0.433, P=0.031), and nodal local efficiency of the right posterior cingulate gyrus was negatively correlated with tongue twister repetition scores (r=-0.422, P=0.036). No significant correlations were observed between global topological metrics and cognitive scores. Lesion volume was negatively correlated only with spatial orientation scores (r=-0.572, P=0.016), and showed no significant correlation with any of the functional network topological metrics.

Patients with subacute ICH exhibit whole-brain functional network reorganization characterized by diminished small-world properties, decreased integration efficiency, and impaired local information processing, suggesting that cognitive impairment after ICH has a broad network basis.

With the increasing frequency of human heat exposure events driven by global climate warming, this study employed the health ecological model to systematically investigate the influencing factors of heat tolerance and their interaction pathways and mechanisms across multiple dimensions including personal traits, behavioral characteristics, environmental adaptation behaviors, and interpersonal networks, aiming to address the risk of heat injury in high-temperature environments.

A cross-sectional survey was conducted on 2 596 university faculty and students in Chongqing from August 21 to September 13, 2024, via the Wenjuanxing and WeChat platforms. Based on the survey data, heat tolerance was classified into 3 levels (low, moderate, strong). Chi-square test, multinomial logistic regression, and structural equation modeling were applied to analyze group differences, independent effects, and path relationships of demographic characteristics and factors at each level of the health ecological model on heat tolerance.

Heat tolerance exhibited group differences with sociodemographic characteristics (sex, age, heatwave experience), personal traits, behavioral characteristics, environmental adaptation behaviors (shower frequency, wearing hats outdoors), and interpersonal networks (weather monitoring, timely seeking help for heatstroke)（P<0.05）. Specifically, females demonstrated significantly lower probabilities of achieving moderate heat tolerance (OR=0.496, 95%CI: 0.390 to 0.631) and strong heat tolerance (OR=0.250, 95%CI: 0.178 to 0.349) compared to males (P<0.001). The participants aged 16 to 18 years (OR=3.778, 95%CI: 2.028 to 7.039) and 19 to 21 years (OR=1.913, 95%CI: 1.021 to 3.586) were more likely to attain strong heat tolerance than those aged ≥26 years（P<0.05）, and those without heatwave experience showed significantly reduced probability of achieving strong heat tolerance (OR=0.475, 95%CI: 0.257 to 0.878,P<0.05). Structural equation modeling further revealed the multipath interactions among these factors. The male advantage in heat tolerance was manifested not only through direct physiological effects (standardized path coefficient=0.261) but also indirectly via enhancement of personal traits (standardized path coefficient=0.185). Although increasing age exerted a direct negative effect on heat tolerance (standardized path coefficient=-0.067), its promotion of personal traits (standardized path coefficient=0.038) indirectly offset this adverse effect. Personal traits emerged as the core determinant of heat tolerance, with a standardized path coefficient as high as 0.696, substantially exceeding all other factors, and serving as a critical mediating variable regulating the overall effects of sex, age, and behavioral characteristics on heat tolerance. Furthermore, although behavioral characteristics showed no significant direct effect on heat tolerance (P=0.871), they generated significant indirect effects through strengthening personal traits (standardized path coefficient=0.304), whereas environmental adaptation behaviors demonstrated a negative impact (standardized path coefficient=-0.143), suggesting that over-reliance on environmental adaptation may suppress the expression of heat tolerance.

Heat tolerance is influenced by multiple integrated factors, among which personal traits constitute the core determinant exerting significant direct and indirect effects on heat tolerance. Enhancing personal traits represents the key strategy for improving heat tolerance and reducing the risk of heat injury.

With the deepening of global aging and neurological disorders having become the second leading cause of death and primary cause of disability worldwide, the prevention of neurological disorders has emerged as an increasingly critical issue. This study employed a two-sample Mendelian randomization (MR) analysis to evaluate the causal associations between birth weight and risk of 6 common neurological disorders, thereby providing genetic epidemiological evidence for the early origins hypothesis of neurological disorders.

This study utilized publicly available genome-wide association study summary statistics. Genetic variants significantly associated with birth weight were selected as instrumental variables to construct two instrumental variable sets comprising 7-single nucleotide polymorphism (SNP) and 60-SNP, respectively. The two-sample MR outcomes included Alzheimer’s disease (AD), Parkinson’s disease (PD), multiple sclerosis, epilepsy, migraine, and cerebrovascular disease, with all relevant data obtained from publicly available datasets on the OpenGWAS platform. Inverse variance weighting was used as the primary analysis, with robustness validation by MR-Egger, weighted median, simple mode, and weighted mode methods. Heterogeneity and horizontal pleiotropy were assessed using Cochran’s Q test, horizontal pleiotropy test, leave-one-out analysis, and funnel plots.

In the 7-SNP analysis, genetically predicted low birth weight was associated with an increased risk of multiple sclerosis, with an OR of 1.323 (95%CI: 1.011 to 1.732, P=0.041) obtained using inverse variance weighting. No significant causal relationships were observed for the remaining 5 neurological disorders (AD, PD, migraine, cerebrovascular disease, and epilepsy). In the 60-SNP analysis, the findings for multiple sclerosis were consistent in direction with the aforementioned association and remained statistically significant, yielding an OR value of 1.319 (95%CI: 1.006 to 1.731, P=0.045) by inverse variance weighting. Although PD showed negative association effects in the weighted median and weighted mode methods, the results of inverse variance weighting did not reach significance (OR=0.811, 95%CI: 0.608 to 1.081, P=0.154). There were no significant causal relationships for the remaining 4 neurological disorders (AD, cerebrovascular disease, epilepsy, and migraine). Sensitivity analyses for multiple sclerosis did not indicate significant systematic pleiotropy in either the 7-SNP and 60-SNP models; heterogeneity increased after expanding from 7-SNP to 60-SNP, but leave-one-out analysis demonstrated that the pooled effect was not driven by any single SNP.

Genetic evidence supports a potential causal relationship between low birth weight and an increased risk of multiple sclerosis, whereas no clear causal relationships are identified with AD, epilepsy, migraine, PD, or cerebrovascular disease.

As a critical support component of the operational system, military logistics and equipment support personnel play a pivotal role in the generation and maintenance of combat force, with their health literacy levels directly influencing these outcomes. However, research focusing on the health literacy of this specific population remains lacking. This study aimed to investigate the current status of health literacy and its influencing factors among logistics and equipment support personnel, thereby providing empirical evidence for constructing a health management system.

A cross-sectional study design was adopted. From April 15 to 30, 2025, 722 logistics and equipment support personnel stationed in a city in western China were selected by cluster sampling. Military Personnel Health Literacy Questionnaire was employed for on-site assessments, encompassing 4 dimensions (basic health concepts, basic health knowledge, basic health skills, and basic health behaviors) and 6 specific categories (military personnel health concepts, prevention of military training injuries, military personnel mental health, combat self-aid and buddy-aid, prevention and control of common diseases in military units, and health management). The questionnaire consists of 40 items with a total score of 50 points. Health literacy was defined as achieving ≥80% of the total score (score ≥40 points). The health literacy rate was calculated using the formula: Health literacy rate = (Number of individuals with adequate health literacy/Total number of valid respondents) × 100%. Spearman analysis was used to examine health literacy-related factors, and multivariable logistic regression was applied to analyze influencing factors for health literacy.

The overall rate of adequate health literacy among the surveyed military logistics and equipment support personnel was 38.14%. Among the 4 dimensions of health literacy, the rate of basic health concept literacy demonstrated the highest level (57.57%), while the rate of basic health knowledge literacy showed the lowest level (22.57%). Among the 6 specific categories of health literacy, the rate of military health concept literacy was as high as 99.57%, whereas the rate of literacy in the prevention and control of common diseases in military units was only 16.00%. The overall correct response rate for health literacy was 77.99%, with 7 items having a correct response rate of less than 60%; among these, the correct response rate for limb fracture fixation techniques was the lowest (31.86%). Multivariate logistic regression analysis revealed that identity category (officer, conscript, etc.) and smoking status were independent influencing factors for health literacy. Compared with conscripts, officers had a significantly higher likelihood of having adequate health literacy (OR=4.960, 95%CI: 1.785 to 13.783). In addition, smokers had a lower level of health literacy than non-smokers (OR=0.662, 95%CI: 0.471 to 0.930).

Health literacy among logistics and equipment support personnel is characterized by strong health concepts, weak combat casualty care skills, and poor competence in the prevention and control of common diseases. It is recommended that future health intervention measures focus on smokers and conscripts, with routine training programs focusing on the prevention and control of military common diseases, military personnel mental health, and combat self-aid and buddy-aid skills.

Elderly patients with metabolic syndrome often present with concurrent subjective cognitive decline and frailty, yet the underlying mechanisms linking these conditions remain unclear. This study aims to explore the parallel mediating effects of physical activity levels and depression on the relationship between subjective cognitive decline and frailty in elderly patients with metabolic syndrome, identify potential pathways and key factors through which subjective cognitive decline influences frailty, and provide evidence to support the development of intervention strategies.

A cross-sectional study was conducted on 348 hospitalized elderly patients with metabolic syndrome admitted to 10 internal medicine departments (including Respiratory Medicine, Neurology, Hypertension and Endocrinology, and Cardiology) in Daping Hospital of Army Medical University from June to December 2024. Subjective cognitive decline was assessed using the Subjective Cognitive Decline Questionnaire (SCD-Q9), depression using the Center for Epidemiologic Studies Depression Scale 10 (CES-D 10), frailty using the Fatigue, Resistance, Ambulation, Illnesses, and Loss of Weight (FRAIL) Scale, and physical activity levels using the International Physical Activity Questionnaire Short Form (IPAQ-SF). Pearson correlation analysis was performed to assess the correlation coefficients between frailty, subjective cognitive decline, depression, and physical activity levels. The Process macro in SPSS 27.0 was used to construct the mediation effect model.

Subjective cognitive decline was positively correlated with depression (r=0.210, P<0.001) and frailty (r=0.275, P<0.001), and negatively correlated with physical activity levels (r=-0.199, P<0.001). Physical activity levels were negatively correlated with frailty (r=-0.228, P<0.001), and frailty was positively correlated with depression (r=0.251, P<0.001). Physical activity levels and depression exerted parallel mediating effects between subjective cognitive decline and frailty, with effect values of 0.009 (95%CI: 0.003 to 0.017) and 0.012 (95%CI: 0.004 to 0.021), accounting for 12.50% and 16.67% of the total effect, respectively.

Physical activity levels and depression play parallel mediating roles between subjective cognitive decline and frailty in elderly patients with metabolic syndrome. Clinicians should emphasize screening for subjective cognitive decline in these patients. And interventions aimed at enhancing physical activity and targeting depressive symptoms may help delay frailty progression and improve quality of life.