Objective To explore the causal relationship between immune cells and heart failure (HF), and the mediating role of serum metabolites, in order to identify potential biomarkers and therapeutic targets. Methods We employed a two-sample Mendelian randomization (MR) analysis method based on genome-wide association study (GWAS) data, analyzing the direct and indirect effects of 731 types of immune cells and 1 400 metabolites on HF. We selected valid instrumental variables and conducted statistical analyses using R software. The primary analysis was performed using the inverse variance weighted method, supplemented by MR-Egger analysis and weighted median method. The stability of the results was assessed through tests such as Cochran’s Q test. Results Our research found a negative causal relationship between PD-L1 on CD14−CD16+ and HF. Sensitivity analysis supported this result. The reverse MR analysis did not find an effect of HF on PD-L1 on CD14−CD16+, indicating that PD-L1 on CD14−CD16+ might play a unidirectional role in reducing the risk of HF. Further mediation MR analysis showed that PD-L1 on CD14−CD16+ might influence the risk of HF onset by regulating the levels of sphingomyelin (d17:1/14:0, d16:1/15:0), with a mediation effect ratio of 6.7%. Conclusion PD-L1 on CD14−CD16+ may reduce the risk of HF by elevating the levels of sphingomyelin (d17:1/14:0, d16:1/15:0), which provides a new perspective for understanding the pathogenesis of HF.

Premature ovarian insufficiency （POI） is a reproductive endocrine disorder that severely affects female reproductive health. However， there remains a lack of clinically effective and safe therapeutic options to restore ovarian function. Traditional Chinese medicine （TCM）， with its advantages of multi-component and multi-target holistic regulation， has shown unique potential in the prevention and treatment of POI. Among TCM monomers， curcumin， resveratrol， and icariin exert antioxidant， anti-inflammatory， and anti-ovarian granulosa cell apoptosis effects by regulating signaling pathways such as nuclear factor-erythroid 2-related factor 2/heme oxygenase-1， phosphoinositide 3-kinase/protein kinase B， and mitogen-activated protein kinase. Single TCM herbs， including American ginseng， Cuscutae Semen， and Cistanches Herba， can regulate sex hormone levels， and alleviate ovarian oxidative stress and immune damage. TCM compound formulas such as Yijing decoction， Siwu decoction， Zuogui pill， and Kuntai capsule modulate multiple signaling pathways， including vascular endothelial growth factor/vascular endothelial growth factor receptor-2， phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin， and silence information regulator 1/p53， thereby promoting angiogenesis， inhibiting mitochondrial apoptosis， and regulating autophagy and ferroptosis， which ultimately delay follicular atresia and improve ovarian reserve function. Further studies are suggested to apply artificial intelligence to deeply explore the underlying mechanisms and carry out high-quality clinical trials， so as to promote the standardized application of TCM in the prevention and treatment of POI.

Objective To investigate the effects and mechanisms of Huatan Quyu Decoction on learning and memory abilities in rats with vascular dementia(VD).Methods Totally 112 male SD rats were randomly selected with 16 rats as the sham-operation group,the remaining rats were used to prepare VD models by segmental ligation of the common carotid artery.The successfully modeled rats were randomly divided into model group,Huatan Quyu Decoction low-,medium-and high-dosage groups(6.1,12.1,24.2 g/kg),donepezil hydrochloride group(0.5 mg/kg)and combination group(Huatan Quyu Decoction 12.1 g/kg+donepezil hydrochloride 0.5 mg/kg),with 16 rats in each group.Each group was given the corresponding treatment measures for 4 weeks.The Morris water maze test was used to assess learning and memory abilities,neurological function was evaluated using Garcia score,HE staining was used to observe the morphology of the hippocampal tissue,ELISA was employed to detect the serum content of Aβ,immunohistochemistry was utilized to observe the β-catenin,LRP6 and GSK-3β protein expression in brain tissue.Results Compared with the sham-operation group,the escape latency of the model group rats was prolonged(P<0.01),the number of crossing platforms was reduced(P<0.01),and the neurological deficit score was decreased(P<0.01),the arrangement of hippocampal tissue cells was disorderly,and the tissue was severely damaged,the serum Aβ content increased(P<0.01),the expressions of β-catenin and LRP6 protein in brain tissue decreased,and the expression of GSK-3β protein increased(P<0.01).Compared with the model group,the escape latency of rats in each administration group was shortened,the number of crossing platforms increased,the neurological deficit score increased,the number of hippocampal cells was relatively more,the arrangement was more orderly,and the structure was relatively complete,the serum Aβ content decreased,the expressions of β-catenin and LRP6 proteins increased,and the expression of GSK-3β protein decreased.Among them,Huatan Quyu Decoction high-dosage group had a significantly better effect than Huatan Quyu Decoction low-and medium-dosage groups(P<0.01),and there was no statistical significance in various indicators compared with the donepezil hydrochloride group(P>0.05).Compared with the donepezil hydrochloride group,the combination group showed significant improvements in learning and memory abilities(P<0.01),the neurological deficit score significantly increased(P<0.01),the number of hippocampal cells significantly increased,arranged neatly,and structurally intact,the serum Aβ content significantly decreased(P<0.01),the expression of β-catenin and LRP6 proteins significantly increased,and the expression of GSK-3β protein significantly decreased(P<0.01).Conclusion Huatan Quyu Decoction can repair cognitive function in VD rats,improve learning and memory abilities,and alleviate VD symptoms by activating the Wnt/β-catenin signaling pathway to reduce serum Aβ content,decrease the apoptosis of nerve cells and alleviate the degree of pathological damage in hippocampal tissue.

Objectives:This study aims to explore the causal relationships between immune cells and serum metabolites in the development of coronary heart disease(CHD)using Mendelian randomization(MR)methods.Methods:We conducted a two-sample MR analysis utilizing data from the FinnGen Project's genome-wide association study(GWAS)on CHD,alongside data on metabolites and immune cells from the GWAS catalog.For the identified associated metabolites,we further employed mediation MR methods to evaluate their mediating roles in the influence of immune cells on CHD.Results:The MR analysis indicated that three immune cells(IgD-CD24-%B cell,CD11b on CD14+monocyte,HLA DR on DC)had stable causal relationship with CHD(under the premise of inverse variance weighted[IVW]method P<0.05,both MR-Egger and weighted median methods also showed P<0.05).Furthermore,these immune cells were associated with CHD through four metabolites(N-acetylneuraminic acid levels,the ratio of desmethyltaurine to taurine,X-24801 and X-18779).Conclusions:This study reveals the causal relationships between immune cells and serum metabolites in the pathogenesis of CHD,providing new biomarkers for early prediction and risk assessment of CHD.

BACKGROUND:The blood-brain barrier is an important structure that protects the central nervous system,and the study of the effects of high glucose on the blood-brain barrier is important for the prevention of high glucose-induced damage to the central nervous system.OBJECTIVE:To investigate the potential effect of high glucose on the blood-brain barrier function of hCMEC/D3 human brain microvascular endothelial cells.METHODS:hCMEC/D3 cells were cultured in regular sugar medium(glucose concentration of 25 mmol/L)and high-sugar medium(glucose concentration of 55 mmol/L).The morphology of cells in each group was observed by light microscopy.CCK-8 assay was used to detect changes in cell viability.A monolayer blood-brain barrier model was established using hCMEC/D3 cell line with Transwell chamber device.Changes in cell transmembrane resistance were monitored daily.The permeability of cell monolayers was detected by phenol red permeability.Flow cytometry was used to detect the apoptosis rate of the cells.Western blot assay was used to detect the expression of Bcl-2,Bax,Caspase-3,ZO-1,Occludin,Claudin-1,and histone deacetylase 4.The levels of histone deacetylase in cell supernatant were detected by ELISA.The expression of histone deacetylase 4 in cells was detected by immunofluorescence.RESULTS AND CONCLUSION:(1)The cell viability of high sugar group was significantly lower than that of control group(P＜0.000 1).(2)The cells of the control group were in a good growth state,interwoven into a dense mesh,with interconnections between synapses.The cell growth of high glucose group was suppressed,and the connection of inter-cellular synapses was reduced.(3)Compared with the control group,the transmembrane resistance value of the high glucose group was reduced(P＜0.05);phenol-red permeability of the monolayer cell membrane was increased(P＜0.05);cell apoptosis rate was increased(P＜0.01);the expression of Bax protein was increased(P＜0.000 1);the expression of Caspase-3 protein had no significant change(P＞0.05);the expression of Bcl-2,ZO-1,Occludin,Claudin-1,and histone deacetylase 4 proteins was decreased(P＜0.01,P＜0.001,P＜0.01,P＜0.000 1,P＜0.01);the fluorescence expression of histone deacetylase 4 was decreased(P＜0.001)in the high glucose group.(4)The level of histone deacetylase 4 in the cell supernatant of the high glucose group was lower than that of the control group(P＜0.05).The results show that high glucose induces the increased apoptosis and enhances permeability of hCEMCE/D3 cells,and its mechanism may be related to the decreased expression level of histone deacetylase 4.

Objective To investigate the relationship between soluble CD73(sCD73) and miRNA(miR)-124a expression level and the severity of acute septic shock and its predictive value for 28 days. Methods A prospective analysis was conducted on 109 patients with acute septic shock admitted to the Third People's Hospital of Chengdu from June 2021 to December 2023. These patients were divided into a survival group (n=87) and a death group (n=22) based on their 28-day prognosis. The clinical data,serum sCD73,miR-124a levels of the two groups were compared,Pearson correlation analysis was used to analyze the relationship between sCD73,miR-124a and acute physiology and chronic health evaluation system Ⅱ (APACHE Ⅱ),sequential organ failure assessment (SOFA),C-reactive protein (CRP) and procalcitonin (PCT). The predictive value of sCD73 and miR-124a for prognosis was analyzed using receiver operating characteristic (ROC) curve analysis,and the interaction between sCD73 and miR-124a in the prognosis of patients with acute septic shock was analyzed using the interaction coefficient γ. Results The age,APACHE Ⅱ,SOFA score,CRP and PCT levels of the death group were higher than that of the survival group(t=4.485～14.147),and the duration of mechanical ventilation was longer than that of the survival group(t=7.215),with statistically significant differences (all P<0.001),respectively. Serum sCD73(3.69±1.12 μg/L) levels were lower in the death group than in the survival group(6.42±2.38 μg/L),and miR-124a (3.31±0.70) was higher than in the survival group(2.63±0.52),and the differences were statistically significant (t=5.222,5.089,all P<0.001). Serum sCD73 and APACHE Ⅱ,SOFA score,CRP and PCT was negatively correlated (r=-0.712～-0.649,all P<0.001),miR-124a and APACHE Ⅱ,SOFA score,CRP and PCT was positively correlated (r=0.643～0.728,all P<0.001). Serum sCD73,miR-124a was significantly correlated with the prognosis of patients with acute septic shock after 28 days of treatment (partial correlation coefficient-0.882,0.896,all P<0.001). The AUC(95％CI) of serum sCD73 and miR-124a for predicting the prognosis of 28-day treatment were 0.765 (0.674～0.841) and 0.754 (0.662～0.831),respectively. The AUC(95％CI) of the combined prediction of 28-day treatment prognosis was 0.916 (0.847～0.960),which was greater than that of the single indicator,and the differences were statistically significant (Z=4.352,4.221,all P<0.001). Low expression of sCD73 and high expression of miR-124a showed a positive interaction in the 28-day mortality risk of patients with acute septic shock (OR=6.668,95％CI:1.245～35.714,γ=1.133,P<0.024). Conclusion Low sCD73 expression and high miR-124a expression positively interacted with each other in the risk of death at 28-day in patients with acute septic shock,and the combined assay is informative for prognostic prediction.

OBJECTIVE To investigate the effects of the split formulas of Biyuan Heji(BYHJ)on paranasal sinus mucosal in-flammation in rats with acute rhinosinusitis(ARS)based on the pathogenesis of"wind-cold transforming into lung-heat".METHODS Unilateral nasal cavity occlusion combined with nasal dripping of Staphylococcus aureus were performed to establish a rat model of ARS.SD rats were randomly divided into blank,model,BYHJ(wind-cold removal+lung-heat removal),lung-heat removal,wind-cold removal,and positive drug groups,with 6 rats in each group.The rats were treated with the corresponding drugs for 7 d and then the samples were collected.HE staining was used to observe the pathological changes of rat paranasal sinus mucosa tissues,ELISA was employed to determine the levels of interleukin-1β(IL-1β),IL-6,IL-8,IL-9,IL-10,and IL-12 in serum,immunohistochemis-try(IHC)was adopted to measure the protein expression of tumor necrosis factor-α(TNF-α)and intercellular adhesion molecule(ICAM-1)in paranasal sinus mucosa tissues,and Western blot was used to detect the protein expression of phosphorylated p38 mito-gen-activated protein kinase(p38 MAPK),nuclear transcription factor-κB p50(NF-κB p50),and NF-κB p65 in paranasal sinus mucosa tissues.RESULTS The acute sinusitis rat inflammation model was successfully established.Compared with the model group,the water drinking,diet eating,and body weight of rats in the BYHJ group,wind-cold removal,lung-heat removal,and positive drug groups were significantly improved,the aggregation of inflammatory cells in the paranasal sinus mucosal tissue was reduced,and the levels of IL-1β,IL-6,IL-8,IL-9,and IL-12 in the serum were significantly reduced(P<0.01),IL-10 content significantly in-creased(P<0.01),the protein expression of TNF-α,ICAM-1,p38 MAPK,NF-κB p50,and NF-κB p65 in the paranasal sinus mucosa was significantly decreased(P<0.01).The comparison between various traditional Chinese medicine groups showed that the decrease of IL-1β,IL-6,IL-8,IL-9,IL-12,TNF-α,ICAM-1,p38 MAPK,NF-κB p50,and NF-κB p65 and the increase of IL-10 in the BYHJ group were better than those in the split formula groups(P<0.01),and the lung-heat removal group was better than the wind-cold removal group(P<0.01).CONCLUSION BYHJ and its split formulas can effectively inhibit the inflammatory response in rats with ARS.

BACKGROUND:The blood-brain barrier is an important structure that protects the central nervous system,and the study of the effects of high glucose on the blood-brain barrier is important for the prevention of high glucose-induced damage to the central nervous system.OBJECTIVE:To investigate the potential effect of high glucose on the blood-brain barrier function of hCMEC/D3 human brain microvascular endothelial cells.METHODS:hCMEC/D3 cells were cultured in regular sugar medium(glucose concentration of 25 mmol/L)and high-sugar medium(glucose concentration of 55 mmol/L).The morphology of cells in each group was observed by light microscopy.CCK-8 assay was used to detect changes in cell viability.A monolayer blood-brain barrier model was established using hCMEC/D3 cell line with Transwell chamber device.Changes in cell transmembrane resistance were monitored daily.The permeability of cell monolayers was detected by phenol red permeability.Flow cytometry was used to detect the apoptosis rate of the cells.Western blot assay was used to detect the expression of Bcl-2,Bax,Caspase-3,ZO-1,Occludin,Claudin-1,and histone deacetylase 4.The levels of histone deacetylase in cell supernatant were detected by ELISA.The expression of histone deacetylase 4 in cells was detected by immunofluorescence.RESULTS AND CONCLUSION:(1)The cell viability of high sugar group was significantly lower than that of control group(P＜0.000 1).(2)The cells of the control group were in a good growth state,interwoven into a dense mesh,with interconnections between synapses.The cell growth of high glucose group was suppressed,and the connection of inter-cellular synapses was reduced.(3)Compared with the control group,the transmembrane resistance value of the high glucose group was reduced(P＜0.05);phenol-red permeability of the monolayer cell membrane was increased(P＜0.05);cell apoptosis rate was increased(P＜0.01);the expression of Bax protein was increased(P＜0.000 1);the expression of Caspase-3 protein had no significant change(P＞0.05);the expression of Bcl-2,ZO-1,Occludin,Claudin-1,and histone deacetylase 4 proteins was decreased(P＜0.01,P＜0.001,P＜0.01,P＜0.000 1,P＜0.01);the fluorescence expression of histone deacetylase 4 was decreased(P＜0.001)in the high glucose group.(4)The level of histone deacetylase 4 in the cell supernatant of the high glucose group was lower than that of the control group(P＜0.05).The results show that high glucose induces the increased apoptosis and enhances permeability of hCEMCE/D3 cells,and its mechanism may be related to the decreased expression level of histone deacetylase 4.

Objective To investigate the relationship between soluble CD73(sCD73) and miRNA(miR)-124a expression level and the severity of acute septic shock and its predictive value for 28 days. Methods A prospective analysis was conducted on 109 patients with acute septic shock admitted to the Third People's Hospital of Chengdu from June 2021 to December 2023. These patients were divided into a survival group (n=87) and a death group (n=22) based on their 28-day prognosis. The clinical data,serum sCD73,miR-124a levels of the two groups were compared,Pearson correlation analysis was used to analyze the relationship between sCD73,miR-124a and acute physiology and chronic health evaluation system Ⅱ (APACHE Ⅱ),sequential organ failure assessment (SOFA),C-reactive protein (CRP) and procalcitonin (PCT). The predictive value of sCD73 and miR-124a for prognosis was analyzed using receiver operating characteristic (ROC) curve analysis,and the interaction between sCD73 and miR-124a in the prognosis of patients with acute septic shock was analyzed using the interaction coefficient γ. Results The age,APACHE Ⅱ,SOFA score,CRP and PCT levels of the death group were higher than that of the survival group(t=4.485～14.147),and the duration of mechanical ventilation was longer than that of the survival group(t=7.215),with statistically significant differences (all P<0.001),respectively. Serum sCD73(3.69±1.12 μg/L) levels were lower in the death group than in the survival group(6.42±2.38 μg/L),and miR-124a (3.31±0.70) was higher than in the survival group(2.63±0.52),and the differences were statistically significant (t=5.222,5.089,all P<0.001). Serum sCD73 and APACHE Ⅱ,SOFA score,CRP and PCT was negatively correlated (r=-0.712～-0.649,all P<0.001),miR-124a and APACHE Ⅱ,SOFA score,CRP and PCT was positively correlated (r=0.643～0.728,all P<0.001). Serum sCD73,miR-124a was significantly correlated with the prognosis of patients with acute septic shock after 28 days of treatment (partial correlation coefficient-0.882,0.896,all P<0.001). The AUC(95％CI) of serum sCD73 and miR-124a for predicting the prognosis of 28-day treatment were 0.765 (0.674～0.841) and 0.754 (0.662～0.831),respectively. The AUC(95％CI) of the combined prediction of 28-day treatment prognosis was 0.916 (0.847～0.960),which was greater than that of the single indicator,and the differences were statistically significant (Z=4.352,4.221,all P<0.001). Low expression of sCD73 and high expression of miR-124a showed a positive interaction in the 28-day mortality risk of patients with acute septic shock (OR=6.668,95％CI:1.245～35.714,γ=1.133,P<0.024). Conclusion Low sCD73 expression and high miR-124a expression positively interacted with each other in the risk of death at 28-day in patients with acute septic shock,and the combined assay is informative for prognostic prediction.

OBJECTIVE To investigate the effects of the split formulas of Biyuan Heji(BYHJ)on paranasal sinus mucosal in-flammation in rats with acute rhinosinusitis(ARS)based on the pathogenesis of"wind-cold transforming into lung-heat".METHODS Unilateral nasal cavity occlusion combined with nasal dripping of Staphylococcus aureus were performed to establish a rat model of ARS.SD rats were randomly divided into blank,model,BYHJ(wind-cold removal+lung-heat removal),lung-heat removal,wind-cold removal,and positive drug groups,with 6 rats in each group.The rats were treated with the corresponding drugs for 7 d and then the samples were collected.HE staining was used to observe the pathological changes of rat paranasal sinus mucosa tissues,ELISA was employed to determine the levels of interleukin-1β(IL-1β),IL-6,IL-8,IL-9,IL-10,and IL-12 in serum,immunohistochemis-try(IHC)was adopted to measure the protein expression of tumor necrosis factor-α(TNF-α)and intercellular adhesion molecule(ICAM-1)in paranasal sinus mucosa tissues,and Western blot was used to detect the protein expression of phosphorylated p38 mito-gen-activated protein kinase(p38 MAPK),nuclear transcription factor-κB p50(NF-κB p50),and NF-κB p65 in paranasal sinus mucosa tissues.RESULTS The acute sinusitis rat inflammation model was successfully established.Compared with the model group,the water drinking,diet eating,and body weight of rats in the BYHJ group,wind-cold removal,lung-heat removal,and positive drug groups were significantly improved,the aggregation of inflammatory cells in the paranasal sinus mucosal tissue was reduced,and the levels of IL-1β,IL-6,IL-8,IL-9,and IL-12 in the serum were significantly reduced(P<0.01),IL-10 content significantly in-creased(P<0.01),the protein expression of TNF-α,ICAM-1,p38 MAPK,NF-κB p50,and NF-κB p65 in the paranasal sinus mucosa was significantly decreased(P<0.01).The comparison between various traditional Chinese medicine groups showed that the decrease of IL-1β,IL-6,IL-8,IL-9,IL-12,TNF-α,ICAM-1,p38 MAPK,NF-κB p50,and NF-κB p65 and the increase of IL-10 in the BYHJ group were better than those in the split formula groups(P<0.01),and the lung-heat removal group was better than the wind-cold removal group(P<0.01).CONCLUSION BYHJ and its split formulas can effectively inhibit the inflammatory response in rats with ARS.