Objective To investigate whether pyroptosis occurs in central post-stroke pain(CPSP)and whether modulation of the NLRP3/GSDMD/IL-1β pathway can alleviate central sensitization. Methods A CPSP model was established by distal middle cerebral artery occlusion via electrocoagulation(dMCAO).C57BL/6J mice were randomly assigned to four groups:Sham+Vehicle,Sham+VX-765,CPSP+Vehicle,and CPSP+VX-765.Protein levels of NLRP3,GSDMD-N,Caspase-1,and IL-1β were measured by Western blotting.Neuronal survival was assessed via Nissl staining.Mechanical and thermal withdrawal threshold of bilateral hind paws were evaluated using von Frey filaments and the Hargreaves test,respectively. Results On day 14 post-modeling,compared with the Sham+Vehicle group,the CPSP+Vehicle group exhibited significantly increased expression of NLRP3,GSDMD-N,Caspase-1 and IL-1β.Neurons around the infarct area showed dissolution and loss of Nissl bodies,along with reduced cell survival.Both mechanical and thermal withdrawal thresholds were significantly decreased.Following VX-765 intervention,the CPSP+VX-765 group displayed decreased expression level of pyroptosis-related proteins,increased neuronal survival around the infarct,and improved withdrawal thresholds. Conclusion VX-765 exerts neuroprotective effects and alleviates central sensitization in CPSP by inhibiting the NLRP3/GSDMD/IL-1β pathway.

Objective To identify core genes influencing immunotherapy response in gastric cancer and to establish a corresponding predictive model. Methods Gene expression data from gastric cancer patients treated with immunotherapy were obtained from the GEO and the TIDE databases.Differentially expressed genes were screened,and weighted gene co-expression network analysis(WGCNA)was performed,identifying 185 genes associated with immunotherapy response for gastric cancer.Core genes were further selected via three machine-learning algorithms:Least Absolute Shrinkage and Selection Operator(LASSO),Random Forest(RF),and Support Vector Machine-Recursive Feature Elimination(SVM-RFE).A predictive model was constructed based on these core genes and validated externally.Model performance was assessed using ROC curves.Additional validation was carried out on an independent dataset GSE62254.Single-cell RNA sequencing was employed to examine the expression of core genes across cell subpopulations. Results A predictive model comprising five core genes was developed,which stratified a cohort of 300 gastric cancer patients into high- and low-risk subgroups.The high-risk group exhibited higher tumor-microenvironment infiltration,TIDE scores,T-cell dysfunction,cancer-associated fibroblast activity,and expression of specific immune-checkpoint molecules,together with lower microsatellite instability scores,collectively indicating enhanced immune escape potential and poorer response to immune checkpoint inhibitors.Single-cell analysis confirmed heterogeneous expression patterns of the five core genes among different cell types. Conclusion A robust predictive model based on differentially expressed core genes related to gastric cancer immunotherapy response was constructed.This model may offer potential value for future research and could aid in clinical decision-making.

Objective To investigate key genes associated with mitophagy and ferroptosis in ischemic stroke(IS)and explore their relationship. Methods Gene expression profiles related to IS were obtained from the Gene Expression Omnibus(GEO),GeneCards,and ferroptosis-related databases.Commonly differentially expressed genes(co-DEGs)were identified,subjecting to functional enrichment and protein-protein interaction(PPI)network analyses.Topological algorithms were applied to screen potential candidate genes,whose diagnostic performance was evaluated by receiver operating characteristic(ROC)curve analysis.The identified target genes were further validated using independent disease-related datasets and animal experiments.Sprague-Dawley (SD)rats were randomly divided into sham(n=10)and model(n=10)groups.Western blotting and quantitative real-time PCR(qRT-PCR)were performed to assess gene expression levels. Results A total of 36 shared genes were identified,which were significantly enriched in pathways associated with autophagy and ferroptosis.Among them,SIRT3,STAT3,HMOX1,and NFE2L2 were identified as key targets.In IS,SIRT3 expression was markedly decreased(P<0.05),whereas STAT3,HMOX1,and NFE2L2 were significantly upregulated(P<0.05).ROC curve analysis yielded area under the curve(AUC)values of 1.0 for all four target genes.Consistent with the bioinformatics findings,animal experiments confirmed increased protein levels of STAT3,HMOX1,and NFE2L2,and decreased SIRT3 in the model group(P<0.05). Conclusion SIRT3,STAT3,HMOX1,and NFE2L2 may serve as critical genes linking mitophagy,ferroptosis,and IS pathogenesis.

Objective To investigate the neuroprotective effect of dexmedetomidine(Dex)on isoflurane-induced postoperative cognitive dysfunction(POCD)in mice and elucidate its underlying mechanism. Methods Fifty SPF-grade aged mice were randomly divided into five groups:control,model,Dex,SFN(Nrf2 activator),and Dex+SFN.Except for the control group,all mice underwent open abdominal surgery under isoflurane anesthesia to establish the POCD model.Cognitive function was assessed using the open field test and Morris water maze test.Hematoxylin-eosin(HE)staining was used to observe morphological changes in hippocampal neurons.TUNEL staining was employed to detect hippocampal neuronal apoptosis.Western blotting was performed to measure the expression levels of Nrf2 and HO-1 proteins in hippocampal tissue. Results Compared with the control group,the model group exhibited significant behavioral inhibition:Reduced total distance traveled[(13.45±1.01)vs.(16.35±1.08)m,P<0.01]and decreased time spent in the center of the open field[(11.05±1.07)vs.(14.21±1.14)s,P<0.01].The model group also showed severe spatial learning and memory impairment:Increased escape latency[(52.34±3.25)vs.(11.45±0.52)s,P<0.01]and decreased number of platform crossings[(2.45±0.13)vs.(8.63±0.41),P<0.01].Degenerative changes were observed in hippocampal neurons,including disordered arrangement,reduced cell count,and structural alterations.The neuronal apoptosis rate was significantly higher in the model group[(51.05 ± 2.58)%vs.(0.12 ± 0.01)%,P<0.01],and Nrf2 and HO-1 protein expression were reduced(both P<0.01).Compared with the model group,all treatment groups exhibited a significant amelioration in behavioral inhibition:The Dex group showed increased distance traveled[(14.50±1.03)m,P<0.05]and increased center time[(12.58±1.08)s,P<0.05];the SFN group also showed increased distance traveled[(14.87±1.02)m,P<0.05]and increased center time[(12.69±1.09)s,P<0.05)].All treatment groups showed significant improvement in cognitive function:The Dex group exhibited decreased latency[(30.43±1.82)s,P<0.01]and increased crossings[(4.85±0.22)times,P<0.01];the SFN group showed decreased latency[(28.74±1.76)s,P<0.01]and increased crossings[(5.12±0.25)times,P<0.01).Hippocampal neuronal damage was markedly ameliorated in all treatment groups.The Dex group had a reduced apoptosis rate[(14.71 ± 1.15)%vs.(51.05 ± 2.58)%,P<0.01];the SFN group also had a reduced apoptosis rate[(18.13 ± 1.32)%vs.(51.05 ± 2.58)%,P<0.01].Both the Dex and SFN group showed increased Nrf2 and HO-1 expressions(P<0.05,P<0.01).Compared with the Dex group,the Dex+SFN group showed further increased distance traveled[(15.68±1.05)m,P<0.05],increased center time[(13.71±1.12)s,P<0.05],decreased latency[(20.03±1.17)s,P<0.01],and increased crossings[(6.74±0.36)times,P<0.01];more significant improvement in hippocampal neuronal damage was observed,along with further reduced apoptosis rate[(10.81 ± 0.94)%vs.(14.71 ± 1.15)%,P<0.01],and further increased Nrf2 and HO-1 expression(both P<0.05). Conclusion Dex exerts a significant neuroprotective effect against isoflurane-induced POCD in mice,which may be associated with the regulation of Nrf2/HO-1 signaling pathway.

Objective To investigate the effects of prenatal alcohol exposure(PAE)on hippocampal GABAergic interneurons in mice. Methods Pregnant C57BL/6 mice were randomly assigned to a pair-fed control(PF-Cont)group or a PAE group.The PAE group received intragastric administration of 4.5 g/kg alcohol from gestational day(GD)5 to 10 and 6.0 g/kg alcohol from GD11 to GD20,while the PF-Cont group received isocaloric maltodextrin solution.On postnatal day(PND)10,spontaneous inhibitory postsynaptic currents(sIPSC)were recorded in hippocampal GABAergic neurons of offspring using the whole-cell patch-clamp technique.Dendritic morphology in hippocampal CA1,CA2/3,and dentate gyrus(DG)regions was quantified via Golgi-Cox staining combined with Sholl analysis.Magnetic resonance imaging(MRI)was performed on male offspring at 72-89 days of age to assess brain morphology.Immunofluorescence staining and stereological methods were used to analyze the density of GABAergic neurons(PV⁺,CR⁺)and total neurons(NeuN⁺)in the hippocampus of male offspring at PND10. Results Male PAE offspring exhibited a significantly higher sIPSC frequency in hippocampal GABAergic neurons than the PF-Cont group(P<0.05).Application of 44 mmol/L alcohol further increased sIPSC frequency in the PAE group(P<0.01),whereas 88 mmol/L alcohol significantly suppressed it(P<0.01).Sholl analysis revealed significantly increased dendritic intersections in PAE male offspring at specific distances from the soma:CA1(10-50 μm),CA2/3(50-140 μm),and DG(100-140 μm)(all P<0.01).Total dendritic length(increased by 31.5% in CA1,38.17% in CA2/3,and 33.2% in DG)and the number of primary dendrites were also significantly elevated(all P<0.05).Adult male PAE offspring showed significantly reduced body weight[(19.64±0.43)vs.(21.32±0.51)g],brain volume[(0.42±0.01)vs.(0.47±0.01)cm³],surface area[(74.81±2.05)vs.(83.05±2.03)mm²],and cortical thickness[(869.54±8.02)vs.(900.32±10.13)μm]compared with the PF-Cont group(all P<0.05).The three-dimensional density of PV⁺ and CR⁺ GABAergic neurons,as well as NeuN⁺ total neurons,was significantly decreased(all P<0.05). Conclusion PAE induces specific impairments of hippocampal GABAergic interneurons in mice,manifesting as aberrant presynaptic excitability(in an alcohol dose-dependent manner),increased dendritic complexity in male offspring,and delayed brain structural development accompanied by reduced density of GABAergic neurons in adulthood.These impairments may constitute a key neurobiological basis for cognitive and behavioral deficits in fetal alcohol spectrum disorders.

Objective To explore the mechanism by which kaempferol inhibits adipose accumulation in obese rats through regulating the expression of insulin receptor(INSR)and peroxisome proliferator-activated receptor γ(PPARG). Methods Experimental rats were randomly divided into the following groups:Control,model,low-dose kaempferol,high-dose kaempferol,and betaine group.Except for the control group,all groups were subjected to obesity induction.The liver index,Lee's index,and the fat-to-body-weight ratio were calculated.Serum levels of lipid profiles(TC,TG,LDL-C,HDL-C),inflammatory factors(TNF-α,IL-6),and liver injury markers(AST,ALT),as well as fasting plasma glucose(FPG),and fasting insulin(FINS)were measured.The homeostasis model assessment of insulin resistance(HOMA-IR)was calculated.Hematoxylin and eosin(HE)staining was performed to assess pathological changes in the liver and adipose tissues.Oil red O staining was used to evaluate hepatic lipid deposition.Immunohistochemistry was performed to detect the expression of CCAAT/enhancer binding protein α(C/EBPα)and acetyl-CoA carboxylase(ACC).Western blotting was used to determine INSR and PPARG protein expression.qRT-PCR was performed to quantify the mRNA levels of C/EBPα,ACC,INSR,and PPARG. Results Compared with the control group,adipocytes in the model group were significantly larger and exhibited marked hypertrophy and hyperplasia,with a disordered arrangement.In the liver tissue,hepatocytes showed extensive vacuolation,increased steatosis,and notable inflammatory cell infiltration;the number and size of lipid droplets were also markedly increased.Furthermore,the liver index,Lee's index,fat-to-body weight ratio,and serum levels of TC,TG,LDL-C,FPG,FINS,HOMA-IR,TNF-α,IL-6,AST,and ALT,were all significantly elevated(all P<0.05).Concurrently,the relative mRNA and protein levels of C/EBPα and ACC,as assessed by qRT-PCR and immunohistochemistry,respectively,were increased(all P<0.05).The relative mRNA and protein levels of PPARG were also upregulated,whereas HDL-C level and the relative mRNA and protein levels of INSR were significantly reduced(all P<0.05).Compared with the model group,the low-dose kaempferol,high-dose kaempferol,and betaine groups showed significant alleviation of pathological damage in adipose and liver tissues,with marked reductions in lipid droplets and lipid accumulation.All measured indices were significantly improved(all P<0.05). Conclusion Kaempferol attenuates adipose accumulation in obese rats,potentially by upregulating INSR expression and downregulating PPARG expression.

Objective To develop predictive models for the risk of gestational diabetes mellitus(GDM)based on targeted metabolomics of serum bile acids. Methods Within a prospective birth cohort,serum levels of 32 bile acids were quantified in early pregnancy.Biomarkers for GDM were screened by the Least Absolute Shrinkage and Selection Operator(LASSO).The eXtreme Gradient Boosting(XGBoost)algorithm was used to build the predictive models,and bootstrap resampling was applied to evaluate model performance.The SHapley Additive exPlanations(SHAP)method was utilized to assess the importance and predictive value of biomarkers. Results The bile acid metabolomics-based models demonstrated strong performance(AUC ≥ 0.840),which outperformed models based on traditional risk factors(AUC=0.828).After incorporating traditional risk factors,predictive performance improved further(AUC ≥ 0.885).Key bile acids identified included glycoursodeoxycholic acid-3-sulfate(GUDCA-3S),taurodeoxycholic acid-3-sulfate(TDCA-3S),taurolithocholic acid(TLCA),glycohyocholic acid(GHCA),ursodeoxycholic acid(UDCA),and the ratios of glycocholic acid to cholic acid(GCA∶CA)and cholic acid to chenodeoxycholic acid(CA∶CDCA).Elevated levels of GUDCA-3S and TDCA-3S,as well as increased GCA∶CA and CA∶CDCA ratios,were associated with higher GDM risk.In contrast,higher concentrations of TLCA,GHCA,and UDCA were associated with lower GDM risk. Conclusion The bile acid metabolomics-based predictive models exhibit robust predictive efficacy.The identified bile acids may serve as potential biomarkers,providing support for early GDM identification,metabolic regulation,and mechanistic research.

Objective To investigate the causal relationships between inflammatory proteins,immune cell-mediated pathways,and osteoporosis using a two-step two-sample Mendelian randomization(MR)approach. Methods This MR study leveraged large-scale genome-wide association studies(GWAS)to analyze 91 inflammatory proteins as exposures,731 immune cell phenotypes as potential mediators,and osteoporosis as the outcome.Inverse variance weighted(IVW)regression served as the primary statistical method,complemented by multilayered sensitivity analyses to enhance robustness. Results The inverse variance weighted method showed that four inflammatory proteins were significantly associated with osteoporosis,all exhibiting positive correlations with disease risk.Among 48 immune cell phenotypes significantly linked to osteoporosis,29 showed positive associations,and 19 showed negative associations with disease risk.Reverse Mendelian randomization analysis revealed no significant correlations,and sensitivity analyses identified no evidence of substantial heterogeneity or horizontal pleiotropy.Furthermore,mediation analysis demonstrated that BAFF-R on IgD^-CD38br partially mediated the causal effect between ARTN(β=0.051,OR=1.053,P=0.002)and osteoporosis.The mediation effect was 0.002,the mediating proportion was 3.9%,and the direct effect was 0.049. Conclusion Mendelian randomization analysis provided supporting evidence for bidirectional causal relationships between inflammatory proteins and osteoporosis,identifying that ARTN may drive osteoporosis development via increasing BAFF-R on IgD^-CD38br.

Objective To investigate the mechanism by which Tongqiao Huoxue decoction(TQHXD) regulates the Caspase-3/Gasdermin E(GSDME)pathway-mediated pyroptosis to improve cognitive function in a rat model of Alzheimer's disease(AD). Methods Rats were randomly divided into the following groups(n=12):Normal group,AD group,TQHXD low-dose group,TQHXD high-dose group,donepezil group,and TQHXD high-dose+Caspase-3 activator(Raptinal)group.Except for the normal group,rats in the other groups received bilateral hippocampal injection of β-amyloid 25-35(Aβ_25-35)to establish the AD model.After successful modeling,treatments were administered once daily for 4 weeks.Cognitive function was assessed using the Morris water maze test;HE staining was used to examine histopathological changes in the hippocampal CA1 region and assess neuropathological scores;the number of Gasdermin E N-terminal fragment(GSDME-N)-positive cells in the hippocampal CA1 region was determined by immunofluorescence staining;levels of interleukin-1β(IL-1β)and IL-18 in the hippocampal CA1 region were measured by ELISA;and the protein levels of cleaved Caspase-3 and GSDME-N in the hippocampal CA1 region were determined by Western blotting. Results Compared with the normal group,the AD group showed disordered neuronal arrangement,nuclear pyknosis,significant inflammatory cell infiltration,increased escape latency,decreased number of platform crossings,and increased neuropathological scores,number of GSDME-N-positive cells,and levels of IL-1β,IL-18,cleaved Caspase-3,and GSDME-N protein in the hippocampal CA1 region(all P<0.05).Compared with the AD group,the TQHXD low-dose group,TQHXD high-dose group,and donepezil group exhibited alleviated pathological damage in the hippocampal CA1 region,decreased escape latency,increased number of platform crossings,and decreased neuropathological scores,number of GSDME-N-positive cells,and levels of IL-1β,IL-18,cleaved Caspase-3,and GSDME-N protein in the hippocampal CA1 region(all P<0.05).Compared with the TQHXD high-dose group,the TQHXD high-dose+Raptinal group showed more severe pathological damage in the hippocampal CA1 region,increased escape latency,decreased number of platform crossings,and increased neuropathological scores,number of GSDME-N-positive cells,and levels of IL-1β,IL-18,cleaved Caspase-3,and GSDME-N protein(all P<0.05). Conclusion TQHXD may inhibit neuronal pyroptosis and neuroinflammation in AD rats by suppressing the Caspase-3/GSDME pathway,thereby improving cognitive function.

Objective To investigate the effects of Bushen Huatan decoction(BSHT) on the β₂ adrenergic receptor(β₂AR)and the cyclic AMP(cAMP)-protein kinase A(PKA)-cAMP response element binding protein(CREB)signaling pathway in ovariectomized rats. Methods A postmenopausal osteoporosis rat model was established by ovariectomy.30 rats were randomly divided into 3 groups:A sham operation (Sham)group,a model (OVX)group,and a (BSHT)group.5 weeks after the operation,the rats received different treatments(4 mL of normal saline for the Sham and OVX groups,and BSHT for the BSHT group).Micro-computed tomography(micro CT)was used to assess the microstructure of the femur.The mRNA and protein expression levels of β₂AR,PKA,and CREB in the femur were detected by RT-PCR and Western blotting,respectively.The distribution of the proteins in femur tissue was determined by immunohistochemistry(IHC). Results Compared with the Sham group,the OVX group showed significantly decreased bone mineral density(BMD),bone volume fraction(BV/TV),trabecular thickness(Tb.Th),and trabecular number(Tb.N),as well as increased trabecular separation(Tb.Sp).The mRNA and protein levels of PKA and CREB were also decreased,while the mRNA and protein levels of β₂AR were significantly increased(all P<0.05)in the OVX group.Compared with the OVX group,the BSHT group exhibited significantly increased BMD,BV/TV,Tb.Th,Tb.N,and expression levels of PKA and CREB,along with significantly decreased expression levels of β₂AR(all P<0.05).IHC results showed that β₂AR,PKA,and CREB were positively expressed in the cytoplasm of osteoblasts surrounding the trabeculae and in bone marrow stromal cells. Conclusion BSHT may prevent and treat osteoporosis in ovariectomized rats by regulating the expression of β₂AR and the cAMP-PKA-CREB signaling pathway.

Objective To investigate the effect of Kangfuxin liquid(KFX)on lipopolysaccharide(LPS)-induced inflammatory injury in oral mucosal epithelial cells(OMECs)and its underlying mechanism. Methods Mouse OMECs(mOMECs)were isolated,identified,and cultured ex vivo,then treated with various concentrations of KFX(0,0.078%,0.156%,0.313%,0.625%,1.25%,2.5%,5%,10%)or LPS(0,0.1,1,10 μg/mL)for 48 h.Cell proliferation and apoptosis were assessed by CCK-8 assay and flow cytometry,respectively,to determine the optimal concentrations of KFX and LPS for intervention.The mOMECs were then divided into the following groups:Control,LPS(1 μg/mL LPS),KFX(0.625% KFX),KFX+negative control inhibitor(KFX+inhibitor-NC),and KFX+miR-449a inhibitor.Cell proliferation was assessed by CCK-8 assay,and apoptosis by flow cytometry.The levels of TNF-α and IL-1β in the supernatant were measured by ELISA.The expression of miR-449a and Notch1 mRNA was determined by qRT-PCR.The protein expression levels of Notch1,cleaved-Caspase-3,Bcl-2,and Bax were detected by Western blotting.Potential binding sites between miR-449a and Notch1 were predicted using bioinformatics tools.The targeting relationship between miR-449a and Notch1 was verified by dual-luciferase reporter assay,qRT-PCR,and Western blotting. Results Compared with the Control group,the LPS group showed significantly decreased cell proliferation,miR-449a expression,and Bcl-2 protein levels(all P<0.05),along with increased apoptosis,TNF-α and IL-1β levels,Notch1 mRNA expression,and Notch1,cleaved-Caspase-3,and Bax protein levels(all P<0.05).Compared with the LPS group,the KFX group exhibited increased cell proliferation,miR-449a expression,and Bcl-2 protein levels(all P<0.05),and decreased apoptosis,TNF-α and IL-1β levels,Notch1 mRNA expression,and Notch1,cleaved-Caspase-3,and Bax protein levels(all P<0.05).Compared with the KFX group,the KFX+miR-449a inhibitor group showed reversed trends in all aforementioned indicators(all P<0.05),whereas no significant changes were observed in the KFX+inhibitor-NC group(P>0.05).Dual-luciferase reporter assay,qRT-PCR,and Western blotting confirmed that miR-449a targeted and inhibited Notch1 expression. Conclusion KFX may ameliorate LPS-induced inflammatory injury in mOMECs via the miR-449a/Notch1 axis.

Objective To explore the mechanism of miR-124 in regulating the PI3K/AKT/NF-κB signaling pathway in insomnia,to open up new ideas for the construction of an insomnia prevention and treatment system. Methods Thirty-two Sprague-Dawley rats were randomly divided into the Con group,SD group,SD miR-124 agomir group,and SD miR-124 antagomir group.HE staining was used to observe the pathological changes of the hippocampus of rats in each group.The ultrastructure of neurons and synapses was observed by transmission electron microscopy.The expression levels of miR-124 in serum and miR-124,PI3K,and AKT in hippocampal tissues were detected by qRT-PCR.ELISA was used to detect the contents of serum interleukins IL-6,IL-1β,and TNF-α in rats,and Western blotting(WB)was used to detect the expression levels of PI3K,AKT,p-AKT,and NF-κB proteins. Results (1)Body weight changes:Compared with the Con group,after modeling,each treatment group showed varying degrees of body weight loss(all P<0.05),with the SD miR-124 antagomir group showing the most significant weight reduction;(2)Sleep phases:EEG/EMG results showed that with increased sleep deprivation time,the duration of WAKE decreased,while the durations of NREM and REM increased relatively(all P<0.05).Compared with the SD group,the SD miR-124 agomir group had a larger increase in REM percentage,and a smaller NREM increase;(3)HE staining:Neuronal death and inflammatory cell infiltration of varying degrees were observed in the CA1,CA3,and DG regions of each treatment group,with the SD miR-124 antagomir group being the most severe,and the SD miR-124 agomir group being milder.Transmission electron microscopy showed mitochondrial swelling,increased autophagosomes,nuclear membrane wrinkling,or endoplasmic reticulum expansion in each treatment group;(4)ELISA results:Compared with the Con group,serum levels of IL-6,TNF-α,and IL-1β increased in the SD group(all P<0.05);compared with the SD group,the SD miR-124 antagomir group had increased inflammatory factors,while the SD miR-124 agomir group showed decreased levels;(5)qRT-PCR results:Compared with the Con group,levels of miR-124,PI3K,and AKT in hippocampal tissue decreased in the SD group;compared with the SD group,PI3K and AKT levels decreased in the SD miR-124 antagomir group,whereas PI3K and AKT levels increased in the SD miR-124 agomir group;(6)WB results:Compared with the Con group,expressions of PI3K and p-AKT proteins in the hippocampus of the SD group significantly decreased(all P<0.05).Compared with the SD group,PI3K and p-AKT levels in the SD miR-124 antagomir group decreased further(all P<0.05),whereas levels of PI3K,AKT,and p-AKT increased in the SD miR-124 agomir group(all P<0.05).Compared with the Con group,NF-κB protein expression increased in the hippocampus of the SD group,SD miR-124 antagomir group,and SD miR-124 agomir group,with the SD miR-124 antagomir group showing the greatest increase. Conclusion This study first predicted the potential association between miR-124 and insomnia,as well as its pathway,through bioinformatics analysis,leading to subsequent experiments to verify that miR-124 can improve the accumulation of inflammatory factors and changes in sleep phases in rats caused by sleep deprivation.This may be related to the regulation of the PI3K/AKT/NF-κB signaling pathway,exerting anti-inflammatory and antioxidant effects.

Objective To investigate the expression of CaMKⅡ in aldosterone-producing adenoma(APA),adrenal nonfunctional adenoma(NFA),and normal adrenal tissue. Methods The mRNA and protein expression of CaMKⅡ in APA,NFA,and normal adrenal tissues was detected by quantitative PCR(qRT-PCR),Western blotting,and immunohistochemical staining. Results Both qRT-PCR and Western blot analysis revealed that the mRNA and protein levels of CaMKⅡ were significantly higher in APA than in normal adrenal glands and NFA(all P<0.05).Immunohistochemical staining in the adrenal cortex demonstrated that CaMKⅡ was primarily detected in the zona glomerulosa(ZG),with weak or negligible expression in the zona fasciculata and reticularis.In APA tissues,CaMKⅡ immunoreactivity was observed in a subpopulation of steroidogenic cells resembling ZG-like cells,particularly located beneath the capsule or surrounding blood vessels.The peritumoral tissue adjacent to APA exhibited significantly more remodeling than that adjacent to NFA and normal adrenal tissue(both P<0.05).Furthermore,CaMKⅡ immunoreactivity score(IRS)was significantly higher in APA tissues than both normal adrenal and NFA tissues(both P<0.05).A positive correlation was found between CaMKⅡ IRS multiplied by tumor volume and supine plasma aldosterone concentration(PAC)(r=0.41,P<0.05),standing PAC(r=0.47,P<0.05),and 24 h urinary aldosterone(r=0.38,P<0.05).No significant correlations were observed with supine plasma renin activity(PRA),orthostatic PRA,or serum K⁺levels. Conclusion CaMKⅡ may be involved in pathological aldosterone synthesis and secretion,as well as adrenal cortex remodeling.

Objective To identify diagnostic biomarkers of plasma extracellular vesicle microRNAs(EV-miRNAs)for acute myocardial infarction(AMI)and to investigate their clinical diagnostic significance. Methods Plasma samples were obtained from three AMI patients and three healthy controls.miRNAs were extracted from both plasma and extracellular vesicles and subsequently subjected to sequencing to establish EV-miRNA profiles.Bioinformatics analysis was performed to select EV-miRNAs significantly associated with AMI.Validation plasma samples were collected from 27 AMI patients and 25 healthy controls for qRT-PCR analysis of the screened differentially expressed EV-miRNAs.The diagnostic performance of AMI-predictive EV-miRNAs was evaluated using ROC curve analysis.Target genes of the selected miRNAs were correlated with clinical indicators. Results Compared with the healthy control group,the expression levels of miR-6809-3p,miR-3938,miR-6815-5p,miR-4322,miR-6717-5p,and miR-6797-5p were significantly elevated in peripheral blood extracellular vesicles of the AMI group(all P<0.05).The area under the ROC curve(AUC)for miR-6815-5p,miR-4322,and miR-6717-5p exceeded 0.9,indicating high diagnostic potential.Furthermore,the expression of creatine kinase MB isoenzyme exhibited a weak positive correlation with miRNA-4322. Conclusion Plasma extracellular vesicle miRNAs may serve as potential molecular biomarkers for AMI diagnosis,providing novel insights for the identification of therapeutic molecular targets for AMI.

Objective To evaluate the diagnostic value of superb microvascular imaging(SMI)combined with serum interleukin-17(IL-17),matrix metalloproteinase-9(MMP-9),and pentraxin-3(PTX3)in assessing disease activity of Takayasu's arteritis(TA). Methods A total of 52 TA patients treated at our hospital between April 2020 and November 2024 were enrolled,including 29 with active disease and 23 with inactive disease.Clinical data were compared between the two groups.Vascular assessments of thickened arterial walls were performed by the Toshiba Aplio 500 color Doppler ultrasound system.Serum levels of IL-17,MMP-9,and PTX3 were detected by enzyme-linked immunosorbent assay.SMI grades and serum biomarker levels were compared between the active and inactive groups.Spearman's rank correlation analysis was used to assess the correlation between SMI grade and serum IL-17,MMP-9,and PTX3 levels.Receiver operating characteristic(ROC)curve analysis was performed to evaluate the individual and combined value of SMI grade and serum biomarkers for disease activity of TA patients. Results No significant differences were observed between the active and inactive groups in terms of age,sex composition,body mass index(BMI),disease duration,clinical symptoms,or carotid artery involvement(all P>0.05).Compared with the inactive group,the active group exhibited significantly higher levels of serum IL-17,MMP-9,and PTX3,as well as a distinct distribution of SMI grade(all P<0.05).Spearman's analysis showed that SMI grade was positively correlated with IL-17,MMP-9,and PTX3 levels(r=0.418,0.502,0.459,all P<0.01).ROC curve analysis showed that the area under the curve(AUC)for SMI grade,serum IL-17,MMP-9,and PTX3 in assessing the disease activity was 0.849,0.723,0.835,and 0.817,respectively.The combined assessment of all four indicators yielded an AUC of 0.949,which was significantly higher than that of any single indicator(Z=2.044,P=0.041;Z=3.061,P=0.002;Z=2.050,P=0.040;Z=2.446,P=0.014),with a sensitivity of 86.21% and specificity of 91.30%. Conclusion SMI grading,alone or combined with serum IL-17,MMP-9,and PTX3,is useful for evaluating disease activity of TA,with enhanced diagnostic performance when used together.

Monkeypox(Mpox)is an emerging zoonotic disease caused by the monkeypox virus(MPXV).The 2022 global outbreak highlighted the pivotal role of vaccination in pandemic control.Currently,no MPXV-specific vaccine is universally approved,therefore,prevention still relies primarily on cross-protective immunity from traditional smallpox vaccines based on the vaccinia virus(VACV),such as ACAM2000,MVA-BN,and LC16m8.This review first evaluates the immunological characteristics,clinical efficacy,and limitations of these conventional vaccines.It then focuses on advances in novel vaccine platforms,including recombinant protein vaccines(e.g.,multivalent antigen fusions and virus-like particles),nucleic acid vaccines(DNA/mRNA),and engineered extracellular vesicle-based vaccines.These platforms are designed to target MPXV-specific antigens rationally,aiming to induce stronger and more durable immune protection.Preclinical studies have shown that multivalent mRNA vaccines(e.g.,BNT166,mRNA-1769)and multivalent VLP vaccines can elicit protection superior to that of the traditional MVA vaccine in animal models.We conclude that future MPXV vaccine development requires continued advances in defining protective correlates,optimizing antigen combinations and delivery systems,and improving accessibility in resource-limited settings.By tracing the shift from cross-protection to multi-platform innovation,this review offers a valuable reference for establishing a scientific framework for Mpox prevention and control.

Inflammatory bowel disease(IBD)is a chronic non-specific inflammatory condition of unknown etiology and poorly understood pathogenesis.Its treatment remains challenging,and the disease was once termed “green cancer”.Patients require long-term or even lifelong treatment,which imposes a substantial burden on both individuals and society.Traditional Chinese medicine has demonstrated efficacy and unique advantages in treating IBD.In recent years,numerous studies have revealed that the gut microbiota plays a key role in the development and progression of IBD.This article reviews the interactions between the gut microbiota and IBD,and explores the current state of research on traditional Chinese medicine for the prevention and treatment of IBD from the perspective of regulating the gut microbiota.We aim to provide new therapeutic ideas and strategies for the clinical management of IBD,and to enable traditional Chinese medicine to play a greater role in future therapeutic strategies and drug development for this disease.

Diabetic cardiomyopathy(DCM)refers to specific structural and functional myocardial abnormalities that develop independent of coronary artery disease,hypertension,or valvular disorders.Although all forms of diabetes can lead to myocardial injury,DCM associated with type 2 diabetes mellitus(T2DM)is the most prevalent and most extensively studied subtype.The onset and progression of DCM involve disturbances in energy metabolism,chronic inflammation,and immune dysregulation.In recent years,the gut microbiota—often referred to as the “second genome” —has been recognized as a pivotal modulator of the pathophysiological processes underlying DCM.This review systematically summarizes the structural and functional alterations of the gut microbiota observed in patients with T2DM and DCM,highlighting its regulatory roles through multiple metabolic and immune pathways,including the bile acid-farnesoid X receptor/Takeda G-protein receptor 5 axis,branched-chain amino acid metabolism-mitochondrial function coupling,trimethylamine N-oxide-mediated inflammatory signaling,the short-chain fatty acids/glucagon-like peptide-1/AMP-activated protein kinase pathway,and the tryptophan-indole-aryl hydrocarbon receptor axis,as well as mechanisms involving gut barrier disruption and endotoxemia.Furthermore,the potential involvement of non-bacterial microbial components—such as fungi,bacteriophages,and enteric viruses—in DCM pathogenesis is discussed,alongside recent advances in microbiota-targeted interventions including probiotics,prebiotics,and fecal microbiota transplantation.Future investigations employing large-scale,multicenter,and longitudinal designs combined with multi-omics analyses are warranted to elucidate the causal relationships and translational potential of the gut-heart axis,which will provide novel insights into the early prediction and precision management of DCM.

Cerebral small vessel disease(CSVD)refers to a group of pathological processes affecting the small cerebral arteries,arterioles,capillaries,and venules of the brain,typically 40-200 μm in diameter.This disease is common in the elderly population and poses a significant threat to their health.Disorder of glucose metabolism is a key independent risk factor for CSVD progression.It can induce vascular endothelial dysfunction,disruption of the blood-brain barrier,and white matter injury through mechanisms such as insulin resistance,chronic inflammatory responses,and oxidative stress,thereby accelerating CSVD.This article reviews research advances on the role of glucose metabolism disorder in CSVD,aiming to provide a theoretical basis for the clinical treatment of CSVD and a reference for understanding its pathogenesis.

Spinal cord injury(SCI)is a devastating disorder of the central nervous system that frequently results in impairment or loss of autonomic,sensory,and motor functions.Although various therapeutic strategies are currently available,their effects are often transient and limited,failing to yield significant clinical outcomes.Transcutaneous spinal cord electrical stimulation(TSCS)is an emerging neuromodulation technique that has become an important non-pharmacological intervention in SCI rehabilitation and neuroscience research.Its advantages-including non-invasiveness,high safety,reversibility,good tolerability,and relatively low cost-have contributed to its significant therapeutic effects on motor function,spasticity,and autonomic nervous function,demonstrating great promise in the field of post-SCI rehabilitation.At present,clinical research on TSCS in China is gradually advancing,however its clinical application still lacks an objective and well-established foundation.This article aims to review the basic principles,potential mechanisms of action,and research progress on the application of TSCS in treating functional disorders after SCI,in order to provide a reference for both clinical practice and future scientific research in this area.

Osteoarthritis is a chronic bone and joint disease characterized primarily by the degenerative changes of articular cartilage.Current clinical treatments primarily focus on delaying disease progression and managing clinical symptoms;however,challenges remain in promoting cartilage regeneration and repair.Recent studies have indicated that pyroptosis plays a significant role in the pathogenesis of osteoarthritis.It exacerbates joint inflammatory responses and cartilage destruction by activating the NLRP3 inflammasome and releasing pro-inflammatory cytokines such as IL-1β.This review systematically elaborates on the molecular mechanisms by which post-translational modifications regulate pyroptosis.It focuses on how modifications such as ubiquitination,phosphorylation,acetylation,and SUMOylation dynamically regulate the assembly of the NLRP3 inflammasome and the activation of Gasdermin proteins,thereby modulating the process of pyroptosis.This aims to provide a new theoretical perspective for a deeper understanding of the pathogenesis of osteoarthritis and its treatment strategies.

As the global population ages at an accelerating pace,the burden of age-related hematological diseases is increasing.Inflammaging,a chronic,systemic,low-grade inflammatory state associated with aging,has been linked to various age-related diseases,including cardiovascular,cerebrovascular,and neurodegenerative disorders.However,its specific mechanistic role in aplastic anemia(AA)—a bone marrow failure—remains systematically unelucidated.This review comprehensively summarizes inflammaging and its underlying mechanisms,with a particular focus on its role in AA.We aim to elucidate how AA contributes to inflammaging in patients through a bone marrow microenvironment-hematopoietic stem cells-immune system triad network,thereby providing a rationale for future AA prevention and treatment.