Objective To explore the relationship between education level and the risk of all-cause mortality in the middle-aged and elderly (≥45 years old) population in China. Methods Using data from five surveys from 2011-2020, years of education, age, gender, marital status, type of household, alcohol consumption status, smoking status, physical activity, limited ability to perform activities of daily living (ADLs), chronic disease status, and body mass index were collected. According to the survival status，the study participants were divided into a survival group (n=8625) and an all-cause mortality group (n=1735). Cox proportional risk regression model was used to analyze the relationship between years of education and the risk of all-cause mortality in middle-aged and elderly people with stratified analysis. Results The survey was conducted on 10360 research subjects, including 4 983 males and 5 377 females, with an age M(Q_R) of 59(8) years old. A total of 88 187 person years (average 8.512 person years) were followed up, and a total of 1735 deaths were reported over 9 years. The all-cause mortality rate was 19.674 ‰, with an education period M(Q_R) of 5(8) years and a survival time M(Q_R) of 9 (0) years. After adjusting for confounding factors, for every 1 year increase in education, the risk of all-cause mortality decreased by 2.60% [HR=0.974, 95% CI (0.960-0.988)]. The stratified analysis results showed that in the population aged 45-59, for every 1 year increase in education, the risk of all-cause mortality decreased by 1.00% [HR=0.990, 95% CI (0.959,1.021)]; In the population aged 60-74 and over 75, for every 1 year increase in education, the risk of all-cause mortality decreased by 2.70% [HR=0.973, 95% CI (0.948, 0.999)] and 3.50% [HR=0.965, 95% CI (0.929, 1.003)], respectively. Conclusion Improving the education level of middle-aged and elderly people (≥ 45 years old) in China can reduce the risk of all-cause mortality, and elderly education should be vigorously promoted.

We have isolated an intestinal probiotic strain, Pediococcus acidilactici HRQ-1. To improve its gastrointestinal fluid tolerance, transportation and storage stability, and slow-release properties, we employed the extrusion method to prepare the microcapsules with P. acidilactici HRQ-1 as the core material and sodium alginate and chitosan as the wall material. The optimal conditions for preparing the microcapsules were determined by single factor and orthogonal tests, and the optimal ratio was determined by taking the embedding rate, survival rate, storage stability, gastrointestinal fluid tolerance, and release rate as the evaluation indexes. The results showed that under the optimal embedding conditions, the embedding rate reached (89.60±0.02)%. Under the optimal formula of freeze-drying protective agent, the freeze-drying survival rate reached (76.42±0.13)%, and the average size of the microcapsules produced was (1.16±0.03) mm. The continuous gastrointestinal fluid simulation experiments confirmed that the microcapsules ensured the viable bacterial count and can slowly release bacteria in the intestinal fluid. The curve of the viable bacterial count during storage at 4 ℃ and room temperature indicated that the prepared microcapsules achieved strains' live number protection. The formula and preparation process of P. acidilactici microcapsules may provide a technological reserve for the preparation of more live bacterial drugs in the future.
Pediococcus acidilactici/chemistry* ; Probiotics/chemistry* ; Capsules/chemistry* ; Alginates/chemistry* ; Chitosan/chemistry* ; Drug Compounding/methods* ; Glucuronic Acid/chemistry* ; Hexuronic Acids/chemistry* ; Freeze Drying

Objective:To investigate the expression of protein kinase D2(PRKD2)in bladder cancer(BLCA)tissue using bioinformatics analysis method and its effect on the prognosis of BLCA patients,and to clarify the role of PRKD2 in the occurrence and development of BLCA.Methods:The data from 9 normal bladder samples,19 BLCA paracancerous samples,and 407 BLCA tumor samples were downloaded from the UCSC Cancer Genome Database.The Mann-Whitney U test was applied to analyze the difference in expression of PRKD2 mRNA in BLCA tumor and normal bladder tissues,and the Human Protein Atlas(HPA)database was used for proteomic validation.DESeq2 package in R software was applied to screen the differentially expressed genes(DEGs)in BLCA tissue in PRKD2 low-and high-expression groups.The co-expression heatmaps of PRKD2 were plotted using the ggplot2 package,Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)were used for functional annotation analysis and pathway enrichment analysis of DEGs,and Gene Set Enrichment Analysis(GSEA)was used to obtain the gene sets that were significantly enriched for DEGs.The BLCA samples were divided into low-and high-expression groups according to the expression level of PRKD2,and the correlations between PRKD2 expression and immune cell infiltration in the BLCA patients were analyzed with GSVA package.The relationship between PRKD2 and prognosis of BLCA patients was further analyzed using the survival package and the survminer package.The PRKD2 gene mutations in BLCA tissue were analyzed using the cBioPortal database.The cystitis,bladder polyp and BLCA tissues were collected,and the expression levels of interleukin-17F(IL-17F)protein in BLCA and control tissues were detected using immunohistochemical staining technique.Results:PRKD2 was highly expressed in a variety of malignant tumors,and the expression levels of PRKD2 mRNA and protein in BLCA tissue were significantly increased compared with those in normal bladder tissue(P＜0.05).Single gene differential analysis of PRKD2 yielded a total of 1 058 DEGs,of which a total of 29 genes were up-regulated and 1 029 were down-regulated.The results of GO functional enrichment analysis showed that DEGs were mainly enriched in the biological process(BP),such as chemical stimuli involved in sensory perception,Cajal body,and endopeptidase inhibitor activity.The results of KEGG pathway analysis showed that DEGs were mainly enriched in the pathway of Staphylococcus aureus infection and the pathway of maturity onset diabetes of the young.GSEA analysis showed that DEGs were mainly enriched in the Notch signaling pathway,the retinoic acid-inducible gene-Ⅰ(RIG-Ⅰ)-like receptor signaling pathway,the cytoplasmic DNA screening pathway,the base excision repair signaling pathway,natural killer(NK)cell-mediated cytotoxicity signaling pathway and T cell receptor signaling pathway.The results of immune infiltration analysis indicated that the expression of PRKD2 was positively correlated with five types of cells,such as activated dendritic cells(aDC),NK CD56dim cells and central memory T cells(Tcm)(P＜0.05),and negatively correlated with three types of immune cells,including macrophages,effector memory T cells(Tem)and plasmacytoid dendritic cells(pDC)(P＜0.05).The clinical characteristic subgroup analysis results showed that the expression levels of PRKD2 mRNA in BLCA patients who were over 70 years old and developed lymphovascular invasion were decreased(P＜0.05);the overall survival(OS),disease-specific survival(DSS)and progression-free interval(PFI)in the BLCA patients with PRKD2 high expression were significantly longer than those with PRKD2 low expression(P＜0.05).The univariate and multivariate Cox analyses indicated that distant metastasis,primary therapy outcome and clinicopathologic stage were the important factors affecting BLCA prognosis.About 9％patients had PRKD2 gene mutations,including missense mutation,gene amplification,mRNA low or high expression,and multi-motif mutation.The immunohistochemistry results showed that the expression level of IL-17F protein in BLCA tissue was significantly higher than that in cystitis tissue(P＜0.05).Conclusion:The expression level of PRKD2 in BLCA tissue is obviously increased,which could up-regulate the expression of IL-17F protein,and the decrease of PRKD2 protein expression may be a potential factor for the poor prognosis of BLCA patients.

Aim To investigate the effect and potential mechanisms of tetramethylpyrazine(TMP)on athero-sclerotic plaques.Methods 43 ApoE-/-mice were used to establish the animal model of atherosclerosis(As)by high-fat diet for 8 weeks,3 of which were used for model outcome verification,and another 40 model mice were randomly divided into model group,TMP low dose(25 mg/kg)group,TMP medium dose(50 mg/kg)group,TMP high dose(100 mg/kg)group and atorvastatin(AT,2.6 mg/kg)group,with 8 mice in each group;another 8 C57BL/6J mice were set as control group.After gavaging administration for 8 weeks,the levels of total cholesterol(TC),triglyceride(TG),low density lipoprotein(LDL),high density lipoprotein(HDL)in the serum were detected by biochemical methods,and the As index was calculated.The levels of oxidized low density lipoprotein(ox-LDL),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),monocyte chemoattractant protein-1(MCP-1),intercellular adhesion molecule-1(ICAM-1)in serum were detected by ELISA method.The pathological changes of aorta was evaluated by HE staining.The aortic plaque formation was evaluated by oil red O staining,and the plaque area percentage was calculated.The aortic fibrosis was evaluated by Masson staining,and the collagen area percentage was calculated.The expression of monocyte macro-phage antibody-2(MOMA-2)and α-smooth muscle actin(α-SMA)in the aorta was detected by immunohistochemistry(IHC)method,and the plaque vulnerability index was calculated.The mRNA and protein expression of peroxisome pro-liferator-activated receptor γ(PPARγ),nuclear factor-κB p65(NF-κB p65)in aorta were detected by RT-qPCR or West-ern blot method.Results Compared with control group,the levels of TC,TG,LDL,ox-LDL,TNF-α,IL-1β,MCP-1,ICAM-1 in serum and As index of the mice were significantly increased in model group,while the level of HDL was sig-nificantly decreased(P＜0.05).The aorta showed pathological changes such as uneven thickening of the intima,accu-mulation of foam cells and fat cells,formation of a large number of plaques,lumen stenosis and infiltration of inflammatory cells;the percentage of aortic plaque area,percentage of collagen area,MOMA-2 and α-SMA positive area,plaque vulner-ability index were all significantly increased(P＜0.05).The mRNA and protein expression of PPARγ in aorta were sig-nificantly decreased,and the mRNA and protein expression of NF-κB p65 were significantly increased(P＜0.05).Com-pared with model group,the levels of TC,TG,LDL,ox-LDL,TNF-α,IL-1β,MCP-1,ICAM-1 in serum and As index of the mice were significantly decreased in the TMP medium,high dose group and AT group,the level of HDL was signifi-cantly increased(P＜0.05).The pathological changes of aorta were significantly improved.The plaque area percentage,collagen area percentage,MOMA-2 positive area percentage and plaque vulnerability index were significantly decreased,and the α-SMA positive area percentage was significantly increased(P＜0.05).The mRNA and protein ex-pression of PPARγ in aorta were significantly increased,the mRNA and protein expression of NF-κB p65 were significantly decreased(P＜0.05).Moreover,TMP exhibited a certain dose-dependent effect on various detection indicators(except MCP-1)in As mice.The regulatory effect of TMP high dose group on various detection indicators(except LDL and As index)in As mice was comparable or superior to those of AT group.Conclusion TMP can reduce the area of As plaque and improve the stability of vulnerable plaque in As mice,its mechanism may be related to regulating PPARγ/NF-κB signaling pathway,improving lipid metabolism and inhibiting inflammatory response.

Objective To describe the clinical features of patients with primary sclerosing cholangitis(PSC)in China based on a nationwide multicenter patient cohort,and to investigate the risk factors for prognosis.Methods A retrospective cohort study was conducted among the patients with a confirmed diagnosis of PSC based on the electronic medical record system of seven grade A tertiary hospitals across the country,and related data were extracted.The Mann-Whitney U test was used for comparison of continuous data between groups,and the chi-square test was used for comparison of categorical data between groups.The Kaplan-Meier method was used to estimate liver transplant-free survival,and the log-rank test was used for comparison of survival rate between PSC patients with different features.The Cox regression model was used to identify independent risk factors for the prognosis of PSC patients and the interactions between key factors.Results A total of 107 patients were enrolled,among whom 55.6%(55/99)had large-duct PSC and 29.0%(31/107)had comorbidity with inflammatory bowel disease(IBD).The positivity rate of anti-neutrophil cytoplasmic antibody(ANCA)was 32.9%(24/73),and 50.0%(40/80)of the patients had an increase in IgG/IgM.The median symptom-to-diagnosis interval was 1 year(<1-4.0),and 38.3%(41/107)of the patients had progressed to decompensated cirrhosis at the time of diagnosis.The median liver transplant-free survival time was 114 months(95%confidence interval[CI]:62-166),with a 5-year survival rate of 65.7%.The multivariate analysis showed that an increase in total bile acid(TBA)(hazard ratio[HR]=1.006,95%CI:1.002-1.010,P=0.001)and a prolonged symptom-to-diagnosis interval(HR=1.252,95%CI:1.059-1.480,P=0.009)were independent risk factors for prognosis.The interaction analysis showed that compared with the female patients with TBA<50 μmol/L,both male and female patients with TBA≥50 μmol/L had a significant increase in the risk of liver transplantation or death(male:HR=16.563,95%CI:2.103-130.449,P<0.001;female:HR=17.009,95%CI:2.113-136.934,P<0.001),and compared with the patients with an age of<45 years and a TBA level of<50 μmol/L,the patients with an age of≥45 years and a TBA level of≥50 μmol/L had a significant increase in the risk of liver transplantation or death(HR=10.729,95%CI:1.325-86.859,P=0.026).Compared with the female patients with an symptom-to-diagnosis interval of≤2 years,the male patients with a symptom-to-diagnosis interval of>2 years had an increased risk of liver transplantation or death(HR=4.825,95%CI:1.725-13.644,P=0.003),and compared with the patients with an age of<45 years and a symptom-to-diagnosis interval of≤2 years,the patients with an age of<45 years and a symptom-to-diagnosis interval of>2 years had an increased risk of liver transplantation or death(HR=4.983,95%CI:1.366-18.173,P=0.015).Conclusion Compared with the reports from Western countries,large-duct PSC is also the main type of PSC in China,but with a relatively low proportion,and there is also a relatively low proportion of patients with IBD or positive ANCA.An increase in TBA and a prolonged symptom-to-diagnosis interval are independent risk factors for prognosis,with significant interactions with age and sex.This suggests that early screening and intervention should be enhanced to improve prognosis.

Objective To explore the predictive value of serum procalcitonin(PCT),peripheral blood complement C3,C4 and lymphocyte subsets CD3+,CD4+and CD8+cells for plastic bronchitis(PB)in children with refractory Mycoplasma pneumoniae pneumonia(RMPP).Methods A total of 222 children with RMPP were selected and divided into the PB group(51 cases)and the non-PB group(171 cases)based on whether they were complicated with PB.The serum PCT level of the children at admission was detected by fluorescence immunoquantitative analyzer,and levels of complement C3 and C4 were detected by immunoturbidimetry.The proportions of CD3+,CD4+and CD8+T lymphocytes were detected by flow cytometry.Multivariate Logistic regression analysis was conducted to analyze influencing factors of PB in children with RMPP.Receiver operating characteristic(ROC)curve was used to analyze the predictive value of the indicators.Results The levels of PCT and CD8+T cells and the duration of fever were higher in the PB group than those in the non-PB group,while the levels of complement C3 and CD4+T cells were lower than those in the non-PB group(P＜0.05).Elevated levels of PCT and CD8+T cells were risk factors for the occurrence of PB in children with RMPP,while elevated levels of complement C3 and CD4+T cells were protective factors for the occurrence of PB in children with RMPP(P＜0.05).The AUCs of PCT,complement C3,the proportion of CD4+T cells and the proportion of CD8+T cells predicted separately were 0.763,0.802,0.788 and 0.802 respectively,and the AUC of combined prediction was 0.915.The AUCs of individual predictions were all lower than those of the combined AUC(Z=3.199,2.825,3.112 and 2.514,P＜0.05).Conclusion PCT,complement C3,CD4+T cell ratio,and CD8+T cell ratio are influencing factors for the occurrence of PB in children with RMPP,and their combined detection is beneficial for the early prediction of PB.

Objective To investigate the relationship of triglyceride-glucose(TyG)index and atherogenic index of plasma(AIP)with positive remodeling of coronary arteries in elderly coro-nary heart disease(CHD)patients with concomitant type 2 diabetes mellitus(T2DM).Methods A total of 120 elderly CHD patients with concomitant T2DM admitted to our department from January 2022 to June 2023 were continuously recruited,and according to the remodeling index,they were divided into positive remodeling group(47 cases)and non-positive remodeling group(73 cases).The clinical data were compared between the two groups.Multivariate logistic analysis was used to identify the risk factors for positive remodeling of coronary artery.Spearman correla-tion analysis was employed to analyze the correlation of TyG and AIP with positive remodeling.ROC curve was plotted to analyze the predictive value of TyG and AIP for the remodeling.Results The positive remodeling group had larger ratio of smoking,higher TG and HbA1c levels,and in-creased TyG index and AIP,but lower HDL-C and blood calcium levels than the non-positive re-modeling group(P＜0.05,P＜0.01).Univariate logistic regression analysis showed that smoking,TG,HDL-C,HbA1c,blood calcium,TyG and AIP were risk factors for positive coronary artery remodeling in elderly patients with CHD and T2DM(P＜0.05,P＜0.01).Multivariate logistic re-gression analysis showed that TyG index(OR=7.253,95％CI:2.458-13.364,P=0.035)and AIP(OR=6.017,95％CI:2.205-12.025,P=0.030)were independent risk factors for positive coronary artery remodeling in elderly patients with CHD and T2DM(P＜0.05).The AUC value of TyG index and AIP in predicting positive coronary artery remodeling in elderly CHD patients with T2DM was 0.783 and 0.766,respectively,and the value of their combination in the prediction was 0.868,which was significantly better than the indicator alone(P＜0.05).Conclusion TyG in-dex and AIP are closely associated with positive coronary artery remodeling in elderly CHD pa-tients with T2DM,and can be used as effective indicators in the prediction of positive remodeling,which is of great significance for early clinical identification of high-risk patients and formulation of individual intervention strategies.

Objective:To analyze the clinical and genetic features of a child with Primary ciliary dyskinesia (PCD) due to compound heterozygous variants of the CCDC39 gene and a 22q11.21 deletion, and to explore the potential role of the two types of variants in the formation of complex phenotypes. Methods:A child presented at the Shanxi Children′s Hospital in March 2025. due to multiple congenital anomalies was selected as the study subject. Peripheral blood samples were taken from the child and her parents and subjected to whole-exome sequencing (WES). Candidate variants were verified by Sanger sequencing. Effect of splicing variant was predicted using SpliceAI, and pathogenicity was assessed based on the ACMG guidelines. Copy number variation (CNV) analysis was also performed. This study has been approved by the Medical Ethics Committee of the Hospital (Ethics No.: IRB-WZ-2025-019).Results:The patient has exhibited multiple features including severe pneumonia, bronchiectasis, localized pulmonary emphysema, scoliosis, tetralogy of Fallot, and atrial septal defect. Genetic testing revealed that she has harbored compound heterozygous variants of the CCDC39 gene, namely c. 1167+ 1G>A and c. 1009A>T, which were inherited from her father and mother, respectively, with the latter being a novel likely pathogenic variant. In addition, a heterozygous deletion of approximately 708 kb at 22q11.21 was detected. Conclusion:The coexistence of CCDC39 gene variants and a 22q11.21 deletion may underlay the development of complex clinical phenotypes in this child.

ObjectiveTo investigate the seroprevalence of Japanese encephalitis virus (JEV) IgG antibodies among the healthy individuals in Huzhou City, Zhejiang Province, so as to provide a scientific basis for the development of prevention and control strategies for Japanese B encephalitis (JE). MethodsA cross-sectional study was conducted from 2023 to 2024. Serum samples were collected from healthy individuals across 12 age groups (<8 months, 8 months‒<2 years, 2‒<4 years, 4‒<6 years, 6‒<8 years, 8‒<10 years, 10‒<15 years, 15‒<20 years, 20‒<30 years, 30‒<40 years, 40‒<50 years, 50‒ years) and tested for JEV IgG antibody to determine positivity rates. ResultsThe overall JEV IgG antibody seroprevalence rate was 57.75 % among the healthy individuals in Huzhou City. The positivity rates for the age groups of <8 months, 8 months‒<2 years, 2‒<4 years, 4‒<6 years, 6‒<8 years, 8‒<10 years, 10‒<15 years, 15‒<20 years, 20‒<30 years, 30‒<40 years, 40‒<50 years, and 50‒ years were 10.14%, 23.76%, 77.31%, 66.88%, 73.87%, 58.33%, 41.67%, 55.06%, 47.00%, 61.00%, 62.00%, and 92.00%, respectively, with a statistically significant difference observed in different age groups (χ²=243.996, P<0.001). Seroprevalence rates also differed significantly between males and females (χ²=9.999, P=0.002). Among subjects vaccinated with 0,1 or 2 doses of live attenuated Japanese encephalitis virus vaccine (JEV-L), the IgG antibody seroprevalence rate was 4.76%, 27.27% and 72.33%, respectively (χ²=108.568, P<0.001). Within the 2 doses group, the IgG antibody seroprevalence rate at <1 year, 1 year, 2 years and 3‒5 years after immunization were 80.85%, 77.36%, 66.67% and 65.45%, respectively, indicating a declining trend with time increased since vaccination (Z=-2.024, P=0.043). ConclusionAdults aged 50‒ years maintain high antibody levels, whereas levels are lower among the age groups of <2 years and 8‒<30 years and wane with time since vaccination in Huzhou City. It is recommended to maintain the routine JE vaccination schedule while exploring booster vaccination strategies for the age groups of 8‒<30 years, in order to enhance population immunity against JEV.

Objective To investigate the current status and influencing factors of anticipatory grief of main caregivers of young and middle-aged patients with advanced liver cancer by cross-sectional survey.Methods A total of 330 caregivers of young and middle-aged patients with advanced liver cancer who were admitted to The Third Affiliated Hospital of Naval Medical University from January 2023 to May 2023 were enrolled as research objects by convenience sampling.A questionnaire survey on general information,anticipatory grief and simple coping style was conducted on these caregivers one day before discharge.Results The score of anticipatory grief of the caregivers was 94.10±22.33.Multiple linear stepwise regression analysis was performed with statistically significant data from the characteristics of the research objects as independent variables and anticipatory grief as the dependent variable,and the result showed that gender,monthly income,educational level,personality self-assessment and coping were main factors affecting anticipatory grief of main caregivers of young and middle-aged patients with advanced liver cancer(all P<0.05).The mean score for positive response was 2.23±0.14,which was negatively associated with anticipatory grief(P=0.041).The mean score for negative response was 1.31±0.24,which was positively associated with anticipatory grief(P<0.001).Conclusion The anticipatory grief of main caregivers of young and middle-aged patients with advanced liver cancer is at a high level.The anticipatory grief is negatively related to positive coping,and positively related to negative coping.The main factors affecting anticipatory grief of main caregivers of young and middle-aged patients with advanced liver cancer are gender,family economic status,educational level,and introverted personality.