Objectives:This study aims to evaluate the causal relationship between plasma proteins and myocardial infarction(MI)using two-sample bidirectional Mendelian randomization(MR)analysis,identify key biomarkers,and validate their expression.Methods:The study utilized publicly available genome-wide association study(GWAS)data of 4 907 plasma proteins as the exposure factor,with single nucleotide polymorphisms(SNPs)as instrumental variables,and four MI datasets as outcomes.Two-sample MR analysis was performed using the inverse variance weighted(IVW)method,complemented by simple model,weighted model,weighted median estimator(WME),and MR-Egger regression methods to assess the causal relationship between exposure factors and outcomes.Venn diagrams and word clouds were used to screen proteins associated with MI as candidate biomarkers.Reverse MR analysis was conducted to evaluate reverse causality.Sensitivity analysis was performed to assess the robustness of the results.Immunohistochemistry(IHC)was used to validate the expression of proteasome activator subunit 1(PSME1)and vacuolar protein sorting 29(VPS29)in the aorta of mice,and enzyme-linked immunosorbent assay(ELISA)was used to verify the expression of PSME1 and VPS29 in plasma from patients with acute myocardial infarction(AMI).Results:The two-sample MR analysis indicated that PSME1 was significantly negatively associated with myocardial infarction in all four datasets,with OR(95%CI)of 0.684(0.557-0.839),0.990(0.987-0.993),0.579(0.448-0.748),and 0.993(0.990-0.996),respectively,with all P<0.001.Similarly,VPS29 also showed a significant negative association with MI in all four datasets,with OR(95%CI)of 0.902(0.862-0.945),0.998(0.997-0.999),0.866(0.808-0.929),and 0.998(0.997-0.999),respectively,with all P<0.001.Reverse MR analysis did not detect reverse causality,and sensitivity analysis confirmed the robustness of the results.IHC results showed significantly reduced expression of PSME1 and VPS29 in the aortas of AMI mice with an atherosclerotic background compared to control mice(both P<0.05).ELISA results indicated significantly lower plasma levels of PSME1 and VPS29 in AMI patients compared to healthy controls(both P<0.05).Conclusions:Higher levels of PSME1 and VPS29 are negatively associated with the risk of MI,suggesting that PSME1 and VPS29 may serve as protective biomarkers for cardiovascular diseases.

Objectives:This study aims to evaluate the causal relationship between plasma proteins and myocardial infarction(MI)using two-sample bidirectional Mendelian randomization(MR)analysis,identify key biomarkers,and validate their expression.Methods:The study utilized publicly available genome-wide association study(GWAS)data of 4 907 plasma proteins as the exposure factor,with single nucleotide polymorphisms(SNPs)as instrumental variables,and four MI datasets as outcomes.Two-sample MR analysis was performed using the inverse variance weighted(IVW)method,complemented by simple model,weighted model,weighted median estimator(WME),and MR-Egger regression methods to assess the causal relationship between exposure factors and outcomes.Venn diagrams and word clouds were used to screen proteins associated with MI as candidate biomarkers.Reverse MR analysis was conducted to evaluate reverse causality.Sensitivity analysis was performed to assess the robustness of the results.Immunohistochemistry(IHC)was used to validate the expression of proteasome activator subunit 1(PSME1)and vacuolar protein sorting 29(VPS29)in the aorta of mice,and enzyme-linked immunosorbent assay(ELISA)was used to verify the expression of PSME1 and VPS29 in plasma from patients with acute myocardial infarction(AMI).Results:The two-sample MR analysis indicated that PSME1 was significantly negatively associated with myocardial infarction in all four datasets,with OR(95%CI)of 0.684(0.557-0.839),0.990(0.987-0.993),0.579(0.448-0.748),and 0.993(0.990-0.996),respectively,with all P<0.001.Similarly,VPS29 also showed a significant negative association with MI in all four datasets,with OR(95%CI)of 0.902(0.862-0.945),0.998(0.997-0.999),0.866(0.808-0.929),and 0.998(0.997-0.999),respectively,with all P<0.001.Reverse MR analysis did not detect reverse causality,and sensitivity analysis confirmed the robustness of the results.IHC results showed significantly reduced expression of PSME1 and VPS29 in the aortas of AMI mice with an atherosclerotic background compared to control mice(both P<0.05).ELISA results indicated significantly lower plasma levels of PSME1 and VPS29 in AMI patients compared to healthy controls(both P<0.05).Conclusions:Higher levels of PSME1 and VPS29 are negatively associated with the risk of MI,suggesting that PSME1 and VPS29 may serve as protective biomarkers for cardiovascular diseases.

Objective To investigate the anti-inflammatory action of Celastrol in the ocular tissues of mice with ex-perimental autoimmune uveitis(EAU)and its effect on microglia polarization.Methods A total of 36 healthy B10.RⅢmice at 6-8 weeks of age were selected and randomly divided into the normal control group,EAU solvent control group and Celastrol intervention group,with 12 mice in each group.The interphotoreceptor retinoid-binding protein(IRBP)161-180 and Freund's complete adjuvant were mixed by thorough emulsification and injected subcutaneously into the bilateral thighs and tails of mice in the EAU solvent control group and the Celastrol intervention group with a total volume of 200 μL and 50 μg IRBP 161-180 in each mouse.On 7-14 days after immunization,mice in the Celastrol intervention group received a daily intraperitoneal injection of 0.5 mg·kg-1 Celastrol,and mice in the EAU solvent control group were injected with an equivalent dose of sterile Phosphate Buffered Saline solution.On the 14th day after immunization,the anterior segment of mice in each group was observed by slit-lamp microscope and Hematoxylin and Eosin(HE)staining of tissue sections was performed;the clinical and histopathological scores of mice in each group were obtained by reference to the Caspi grading standards;immunofluorescence staining was used to observe the activation of microglia in the eyes of mice;Western blot was used to detect the protein expression levels of inducible nitric oxide synthase(iNOS)and arginase-1(Argl)in the reti-na;quantitative real-time PCR was used to detect the relative mRNA expression of inflammatory factors in the retina,such as tumor necrosis factor(TNF)-α,interleukin(IL)-1β and IL-6.GraphPad Prism 9.0 was used for data analysis.Results On the 14th day after immunization,it was observed by the slit-lamp microscope that the anterior segment of mice in the EAU solvent control group was markedly congested with dilated iris blood vessels,corneal edema,and anterior chamber exudation;the inflammation in the anterior segment of mice in the Celastrol intervention group was markedly at-tenuated,and the iris blood vessels were seen to be mildly congested.Compared with the normal control group,the clini-cal scores of mice in the EAU solvent control group and the Celastrol intervention group were significantly elevated(both P＜0.05);the clinical scores of mice in the Celastrol intervention group were lower than those in the EAU solvent control group(P＜0.05).HE staining results showed that on the 14th day after immunization,mice in the EAU solvent control group showed severe retinal folds and detachment with diffuse infiltration of inflammatory cells,while mice in the Celastrol intervention group showed slight structural damage to the retina and a small amount of inflammatory cell infiltration.Com-pared with the normal control group,the histopathological scores of mice in the EAU solvent control group and the Celas-trol intervention group were significantly elevated(both P＜0.05);the histopathological scores of mice in the Celastrol in-tervention group were lower than those in the EAU solvent control group(P＜0.05).The intraocular Iba1+cell densities of mice in the normal control,EAU solvent control and Celastrol intervention groups were(1.00±0.12)％,(36.07± 4.57)％,and(1.83±0.36)％,respectively.Compared with the normal control group,the number of Iba1+cells in the eyes of mice in the EAU solvent control group and the Celastrol intervention group significantly increased(both P＜0.05);compared with the EAU solvent control group,the number of Iba1+cells in the eyes of mice in the Celastrol intervention group was significantly reduced(P＜0.05).Compared with the normal control group,the expression levels of iNOS and Arg1 proteins in the retinas of mice in the EAU solvent control group were significantly elevated(both P＜0.01);compared with the EAU solvent control group,the expression of iNOS protein in the retinas of mice in the Celastrol intervention group was significantly reduced(P＜0.01).Compared with the normal control group,the relative mRNA expressions of TNF-α.IL-1β,and IL-6 in the retinas of mice in the EAU solvent control group was significantly elevated(all P＜0.05);compared with the EAU solvent control group,the relative mRNA expressions of TNF-α,IL-1 β,and IL-6 in the retinas of mice in the Celastrol intervention group significantly decreased(all P＜0.05).Conclusion Celastrol inhibits Ml microglia activation and reduces the production of retinal inflammatory factors TNF-α,IL-1 β and IL-6 in EAU mice,thereby attenuating the in-flammatory reaction.

Objective:To explore the value of serum procalcitonin (PCT), amylase (AMY), albumin (ALB) and lactate dehydrogenase (LDH) in the clinical diagnosis and evaluation of severe acute pancreatitis (SAP).Methods:A total of 70 patients with acute pancreatitis treated in Yancheng First People′s Hospital from January 1, 2020 to December 31, 2022 were enrolled as pancreatitis group. According to disease severity, they were divided into mild group (22 cases) and severe group (48 cases). A total of 70 controls during the same period were enrolled as control group. The general data of all the objects were collected at enrollment. The levels of plasma PCT, AMY, ALB and LDH were detected. The diagnostic value of the above indexes for SAP and their evaluation value for disease severity were analyzed by receiver operating characteristic (ROC) curves.Results:The levels of serum PCT, AMY and LDH in the pancreatitis group were significantly higher than those in the control group: (3.14 ± 0.67) μg/L vs. (0.82 ± 0.21) μg/L, (602.53 ± 199.47) U/L vs. (99.97 ± 30.85) U/L, (767.24 ± 198.73) U/L vs. (423.61 ± 59.19) U/L, P<0.05; while ALB was significantly lower than that in the control group: (33.47 ± 6.98) g/L vs. (45.79 ± 6.12) g/L, P<0.05. ROC curves analysis showed that area under the curve (AUC) values of PCT, AMY, LDH, ALB and combined detection in the diagnosis of acute pancreatitis were 0.783, 0.792, 0.697, 0.732 and 0.915, respectively. The levels of serum PCT and LDH in the mild group were significantly lower than those in the severe group: (2.76 ± 0.44) μg/L vs. (3.59 ± 0.61) μg/L, (507.06 ± 131.67) U/L vs. (848.95 ± 207.79) U/L, P<0.05; while ALB was significantly higher than that in the severe group: (35.39 ± 4.73) g/L vs. (32.64 ± 5.09) g/L, P<0.05. ROC curves analysis showed that the AUC values of PCT, LDH, ALB and combined detection for evaluating disease severity were 0.668, 0.749, 0.741 and 0.959, respectively. The evaluation value of combined detection was significantly higher than that of single index ( P<0.05). Conclusions:The levels of serum PCT, AMY and LDH are abnormally increased, while ALB level is abnormally decreased in patients with acute pancreatitis, and which all can be applied for clinical diagnosis. PCT, LDH and ALB can be applied for disease evaluation.

Objectives:A two-sample,multivariable,bidirectional Mendelian randomization (MR) analysis was conducted to explore the causal relationships between serum micronutrients (including vitamin A,vitamin B6,vitamin B9,vitamin B12,vitamin C,vitamin D,vitamin E,copper,iron,selenium,zinc,calcium,magnesium,and potassium) and coronary atherosclerosis (CA).Methods:Publicly available data from genome-wide association studies in European populations were analyzed,14 serum micronutrients were used as exposure factors and single nucleotide polymorphisms (SNPs) were used as instrumental variables,outcome was defined as CA.A two-sample MR analysis was performed using inverse variance weighted (IVW) method,MR-Egger regression,weighted median estimator (WME),simple model,and weighted model to assess the relationship between each micronutrient and CA.Multivariable MR analysis was used to evaluate the independent impacts of single exposure factors on CA,reverse MR was applied to assess the potential for reverse causality.Sensitivity analysis was conducted using Cochran's Q test,MR-Egger regression,MR-PRESSO,and leave-one-out analysis to evaluate the robustness of the results.Results:The univariable two-sample MR study evidenced the significant associations between serum levels of vitamin A (IVW:OR=0.019,95％CI:0.001-0.451,P=0.014),vitamin B12 (IVW:OR=1.221,95％CI:1.015-1.469,P=0.034),copper (IVW:OR=1.023,95％CI:1.003-1.044,P=0.026),and potassium (IVW:OR=0.837,95％CI:0.714-0.980,P=0.027) with the risk of CA.After adjustment using multivariable MR analysis,independent causal effects on CA were observed for serum levels of vitamin A (IVW:OR=0.016,95％CI:0.001-0.214,P=0.002) and copper (IVW:OR=1.029,95％CI:1.002-1.056,P=0.036).Reverse MR analysis observed a reverse causal relationship between CA risk and serum vitamin A level,though the effect was minimal (IVW:OR=0.999),while no reverse causality was found between CA risk and serum copper level.There was no evidence to support a causal relationship between the remaining serum micronutrients and CA.These findings were robust through extensive sensitivity analyses.Conclusions:The levels of serum vitamin A and copper may relate to the susceptibility of CA in the studied population.

Objectives:A two-sample,multivariable,bidirectional Mendelian randomization (MR) analysis was conducted to explore the causal relationships between serum micronutrients (including vitamin A,vitamin B6,vitamin B9,vitamin B12,vitamin C,vitamin D,vitamin E,copper,iron,selenium,zinc,calcium,magnesium,and potassium) and coronary atherosclerosis (CA).Methods:Publicly available data from genome-wide association studies in European populations were analyzed,14 serum micronutrients were used as exposure factors and single nucleotide polymorphisms (SNPs) were used as instrumental variables,outcome was defined as CA.A two-sample MR analysis was performed using inverse variance weighted (IVW) method,MR-Egger regression,weighted median estimator (WME),simple model,and weighted model to assess the relationship between each micronutrient and CA.Multivariable MR analysis was used to evaluate the independent impacts of single exposure factors on CA,reverse MR was applied to assess the potential for reverse causality.Sensitivity analysis was conducted using Cochran's Q test,MR-Egger regression,MR-PRESSO,and leave-one-out analysis to evaluate the robustness of the results.Results:The univariable two-sample MR study evidenced the significant associations between serum levels of vitamin A (IVW:OR=0.019,95％CI:0.001-0.451,P=0.014),vitamin B12 (IVW:OR=1.221,95％CI:1.015-1.469,P=0.034),copper (IVW:OR=1.023,95％CI:1.003-1.044,P=0.026),and potassium (IVW:OR=0.837,95％CI:0.714-0.980,P=0.027) with the risk of CA.After adjustment using multivariable MR analysis,independent causal effects on CA were observed for serum levels of vitamin A (IVW:OR=0.016,95％CI:0.001-0.214,P=0.002) and copper (IVW:OR=1.029,95％CI:1.002-1.056,P=0.036).Reverse MR analysis observed a reverse causal relationship between CA risk and serum vitamin A level,though the effect was minimal (IVW:OR=0.999),while no reverse causality was found between CA risk and serum copper level.There was no evidence to support a causal relationship between the remaining serum micronutrients and CA.These findings were robust through extensive sensitivity analyses.Conclusions:The levels of serum vitamin A and copper may relate to the susceptibility of CA in the studied population.

Objective To investigate the distribution characteristics and risk factors of intracranial and extracranial aterial lesions in Chinese patients with ischemic stroke . Methods In this multi‐center study ,2 310 continuously inpatients with ischemic stroke diagnosed in 20 stroke screening and prevention project base hospitals from June 2015 to M ay 2016 were enrolled . Carotid ultrasonography and transcranial color‐coded sonography or transcranial Doppler were performed in all patients to confirm the presence of cerebral artery stenosis or occlusion . According to the distribution of lesions ,the subjects were divided into 2 groups :the simple intracranial artery stenosis group and the simple extracranial artery stenosis group . T he difference of risk factors between the two groups was compared . Results Of the 2 310 patients with ischemic stroke ,1 516 ( 65 .6% ) had simple intracranial artery stenosis and 794 ( 34 .4% ) had simple extracranial artery stenosis . T he incidence of anterior circulation artery stenosis was higher in the group of intracranial artery stenosis than that in the extracranial artery stenosis group ( 68 .1% vs 48 .7% , P ＜0 .001) . Posterior circulation artery stenosis and combined anterior with posterior circulation artery stenosis were more common in patients with extracranial artery stenosis group than those in intracranial artery stenosis group ( 36 .4% vs 22 .1% ,14 .9% vs 9 .8% ;all P ＜0 .001) . Univariate analysis of risk factors for stroke showed that patients with intracranial arterial stenosis had a higher prevelence of hypertension , diabetes ,obesity ,and family history of stroke ,and their systolic blood pressure ,diastolic blood pressure , body mass index ( BM I) ,fasting blood‐glucose ,glycosylated hemoglobin ,triacylglycerol ,total cholesterol , and low‐density lipoprotein cholesterol were significantly higher than those in the extracranial arterial stenosis group ( all P ＜ 0 .05 ) . T he proportion of elderly ( ≥ 65 years old ) ,male and smokers in the extracranial arterial stenosis group was significantly higher than that in the intracranial arterial stenosis group ( all P ＜0 .05) . Multivariate logistic regression analysis showed that elderly ( ≥65 years old) ,male , and smoking history were independent risk factors for extracranial arterial stenosis ( OR＝ 2 .012 ,1 .637 , 1 .325 ,respectively ;all P ＜0 .05) . While hypertension ,diabetes ,less physical activity ,and high BM I levels were independent risk factors for simple intracranial arterial disease ( OR ＝ 1 .301 ,1 .252 ,1 .248 ,1 .030 , respectively ;all P ＜0 .05) . Conclusions There are significant differences in the distribution characteristics and risk factors of intracranial and extracranial aterial lesions in patients with ischemic stroke in China .

Objective To analyze the correlation between smoking and occurrence of intracranial artery stenosis.Methods From June 2015 to May 2016,a total of 10 711 inpatients with transient ischemic attack (TIA) or ischemic stroke from 20 basel hospitals of nationwide were enrolled using a cross-sectional study,76 patients with unknown smoking and smoking cessation years were excluded.Finally,a total of 10 635 patients were enrolled.Transcranial color coded sonography and/or transcranial Doppler were used evaluate the intracranial artery stenosis lesions.The basic risk factors for cerebrovascular disease (age,sex,smoking and smoking years,whether smoking cessation and years,hypertension,diabetes,hyperlipidemia,atrial fibrillation,and family history of stroke) were recorded.According to the different smoking years,the smoking years were divided into five groups:non-smoking,smoking time ≤10-year,11 to 20-year,21 to 30-year,and >30-year groups for trend chi square test.According to the different smoking cessation years in the smokers,the smoking cessation years were divided into four groups:non-cessation,cessation time 1 to 10-year,11 to 20-year,and >20-year groups for trend chi square test.The effects of different smoking years and different smoking cessation years on the occurrence of intracranial arterial stenosis were analyzed.Results The incidence of intracranial artery stenosis in the smokers (40.4%[1 433/3 547]) was significantly higher than that in the non-smoking patients (29.4%[2 085/7 088]).There was significant difference (χ2=128.850,P<0.01),and the incidence of cerebral infarction in the smokers (91.6%[3 250/3 547]) was significantly higher than the non-smokers (85.0%[6 027/7 088]).There was significant difference (χ2=92.328,P<0.01).Smoking was an independent risk factor for intracranial artery stenosis (OR,1.603;95%CI 1.456-1.765;P<0.01).With the increase of smoking years,the detection rate of intracranial arterial stenosis increased gradually (trend χ2=115.437,P<0.01).Whether giving up smoking had no significant effect on the incidence of intracranial artery stenosis in patients with ≥20 years of smoking (trend χ2=1.043,P=0.307).Conclusions Smoking is an independent risk factor for affecting intracranial artery stenosis;the risk of disease increases with the number of smoking years.Long-term smokers (≥20 years) cannot reduce the effect on intracranial artery stenosis,even if they give up smoking.

Objective To investigate the effect of serum lipid level on carotid artery stenosis in patients with ischemic cerebrovascular disease.Methods Using a multi-center cross-sectional study,10 711 consecutive inpatients with transient ischemic attack (TIA) or ischemic stroke diagnosed clearly in 20 stroke screening and prevention project base hospitals from June 2015 to May 2016 were enrolled.According to the results of carotid ultrasonography,1 560 patients with extracranial carotid artery stenosis rate≥50% screened were enrolled in the study.They were divided into a severe stenosis group (70%-99%) and a mild-moderate stenosis group (<70%).The distribution of total cholesterol (TC),triacylglycerol (TG),low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) levels in both groups of carotid stenosis patients were analyzed,and the quantitative classification was based on the normal range of serum lipids.The distributions of serum lipid levels in different grades in patients of both groups were compared with the non-parameter test.Results The incidence of dyslipidemia in the severe stenosis group was higher than that in the mild and moderate stenosis group.There was no significant difference between the two groups (54.4%[319/586]vs.48.3%[470/974],P<0.05).Dyslipidemia was an independent risk factor for severe carotid artery stenosis (OR,1.27,95% CI 1.24-1.30,P<0.01).The TC and LDL-C levels in patients of the severe stenosis group were significantly higher than those in the mild-moderate stenosis group (TC:3.98[3.31,4.82]mmol/L vs.3.91[3.31,4.53]mmol/L,LDL-C:2.48[1.86,3.14]vs.2.30[1.79,2.80];all P<0.01).With the increase of TC and LDL-C levels,there was significant differences between the severe stenosis group and the mild-moderate stenosis group (all P<0.05),and the proportions of TC >5.80 mmol/L (7.3%[43/586]vs.0.4%[4/974]) and LDL-C>3.12 mmol/L (26.3%[154/586]vs.10.0%[97/974]) in patients of the severe stenosis group were higher than those in the mild-moderate stenosis group (26.3%[154/586]vs.10.0%[97/974]).Conclusion The high LDL-C and TC levels may increase the incidence of severe carotid artery stenosis or occlusion.

Objective To investigate the differences of distribution characteristics and risk factors of large artery lesions in patients with ischemic cerebrovascular disease in different age groups in order to provide the basis for the prevention and treatment of stroke in different age groups.Methods From June 2015 to May 2016,a total of 10 711 consecutive inpatients with transient ischemic attack (TIA) and ischemic stroke from 20 centers nationwide were enrolled.Each 10 years was used as an age group from 40 years.All the patients were divided into 5 age groups.The differences of the different risk factors for cerebrovascular disease among the 5 groups were compared.All patients were separated by gender.The chi square test was used to compare the incidences of large artery stenosis of the intracranial and external and anterior and posterior circulation,and the number of vascular lesions in the same sex in different age groups.Results (1) The risk factors of elderly patients were mainly hypertension,diabetes mellitus,and atrial fibrillation (χ2=61.938,χ2=13.349,and χ2=55.940;all P<0.01).The smoking history,family history of cerebrovascular disease,and obesity were more frequent among the young and middle-aged people (χ2=131.505,χ2=7.298,and χ2=100.911,all P<0.01).(2) The linear trend chi square test results showed that the proportion of multivessel diseases in female and male extracranial arterial lesions increased gradually with the increase of age.(χ2=54.799,χ2=161.370,all P<0.01).The proportion of multivessel diseases in the intracranial artery in female decreased gradually (χ2=5.328,P=0.021),and that in male did not have obvious trend of change (χ2=0.289,P=0.591).(3) The linear trend chi square test results showed that the incidence of simple intracranial arterial stenosis in female and male intracranial arterial stenosis decreased gradually with the increase of age (χ2=20.090,χ2=42.351,all P<0.01),and the incidence of simple extracranial arterial stenosis increased gradually (χ2=40.311,χ2=90.698,all P<0.01).The incidence of both intracranial and extracranial artery stenoses increased gradually (χ2=12.077,χ2=45.887,all P<0.01).The incidence of simple posterior circulation vascular stenosis increased gradually in female (χ2=16.434,P<0.01),but that did not have obvious trend of change in male (χ2=1.701,P=0.192).The incidence of stenosis of both anterior and posterior arteries in female and male increased gradually (χ2=4.587,P=0.032;χ2=35.156,P<0.01).Conclusions The distribution of atherosclerotic lesions in ischemic cerebrovascular disease of the different age groups was different.No matter female or male patients,the majority of the young and middle-aged patients were intracranial artery lesions,and the elderly patients were mainly extracranial artery lesions.The majority of elderly women had posterior circulation artery lesions.Understanding the characteristics in patients with intracranial arterial lesion in different age groups will help to develop individualized stroke prevention and treatment strategies for the population of different age groups.