Clonorchis sinensis primarily lives in the host liver and biliary system, leading to serious health issues, such as hepatobiliary lesions and cholangiocarcinoma. Establishment of rapid, sensitive and specific tools is of great significance for early diagnosis and management of clonorchiasis. Recently, molecular biological techniques have been widely used for detection of C. sinensis infections. This review summarizes the advances in the major molecular assays for detection of C. sinensis infections in intermediate and definite hosts, including PCR, loop-mediated isothermal amplification, clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated proteins system, and high-throughput sequencing, and summarizes the characteristics and application prospects of these assays, so as to provide precision detection and management of C. sinensis infection.

ObjectiveTo construct a machine learning model combining two-dimensional shear wave elastography （2D-SWE） and serological markers， and to investigate its clinical value in differentiating primary biliary cholangitis （PBC） from overlap syndrome （OS）. MethodsA total of 199 patients with pathologically confirmed PBC or OS in Nantong Third People’s Hospital from January 2021 to December 2025 were retrospectively enrolled， with 125 patients in the PBC group and 74 in the OS group. The patients were randomly divided into a training set with 139 patients and a test set with 60 patients at a ratio of 7∶3. Related data were collected， including serological markers， conventional two-dimensional ultrasound parameters， and 2D-SWE parameters （including velocity of shear wave ［VS］ and liver fibrosis index ［LFI］）. The independent-samples t test was used for comparison of normally distributed continuous data between two groups， and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups； the chi-square test was used for comparison of categorical data between two groups. The univariate and multivariate Logistic regression analyses were used to identify independent predictive factors， which were then incorporated into six machine learning models， and 5-fold cross-validation was used for optimization of parameters. The indicators including the area under the receiver operating characteristic curve （AUC） were compared in the test set to determine the best model， and the SHapley additive exPlanations （SHAP） analysis was used for model interpretation. ResultsFemale patients accounted for 87.8% in the OS group and 79.2% in the PBC group. Compared with the PBC group， the OS group had significantly higher age， VS， LFI， splenic area， aspartate aminotransferase， total bilirubin， prothrombin time， immunoglobulin G （IgG）， and immunoglobulin M （Z=-2.883， -6.524， -4.000， -3.061， -2.194， -2.372， -4.079， -6.964， and -2.709， all P<0.05） and a significantly lower platelet count （Z=4.098， P<0.001）， and there were also significant differences between the two groups in the proportion of patients with positive anti-nuclear antibody， fibrosis stage， and inflammation grade （χ²=3.458， 63.198， and 101.038， all P<0.05）. Variables without multicollinearity were included in the regression analysis， and the multivariate Logistic regression analysis showed that VS （odds ratio ［OR］=4.503， 95% confidence interval ［CI］： 1.698 — 11.943， P=0.003）， LFI （OR=1.813， 95%CI： 1.050 — 3.132， P=0.033）， and IgG （OR=1.121， 95%CI： 1.026 — 1.226， P=0.012） were independent predictive factors for differentiating PBC from OS. Six machine learning models were constructed based on these variables， among which the logistic regression model showed the best predictive performance in the test set， with an AUC of 0.881 （95%CI： 0.788 — 0.974）， a sensitivity of 0.731， and a specificity of 0.794. The SHAP analysis showed that IgG contributed the most to model prediction. ConclusionThe noninvasive multimodal machine learning model combining 2D-SWE parameters （VS， LFI） and serum IgG can effectively differentiate PBC from OS， providing a reference for clinical decision-making regarding the need for liver biopsy.

Aim To explore the relationship between serum levels of proprotein convertase subtilisin/kexin type 9(PCSK9),macrophage migration inhibitory factor(MIF)and the severity of coronary artery lesions in patients with coro-nary heart disease(CHD).Methods A cross-sectional study was conducted involving 139 patients with CHD and 69 control subjects who underwent coronary angiography during the same period,all of whom were admitted to the People's Hospital of Xinjiang Uygur Autonomous Region from November 2023 to May 2024.Clinical data and coronary angiography results were collected,and the severity of coronary artery stenosis was quantitatively assessed using the Gensini score.Pa-tients with the Gensini scores＞0 were classified into three groups based on tertiles:the mild stenosis group(1～18 points,54 cases),the moderate stenosis group(19～36 points,54 cases),and the severe stenosis group(＞36 points,54 ca-ses).Serum levels of PCSK9 and MIF were measured by ELISA kit.Results Serum levels of PCSK9 and MIF were significantly higher in the CHD group than those in the control group(P＜0.05).Multivariable Logistic regression analy-sis revealed that high levels of serum PCSK9 and MIF were independent risk factors for CHD.Spearman correlation analy-sis showed that serum PCSK9 and MIF levels were positively correlated with Gensini score(rs=0.619 6 and r,=0.411 4,both P＜0.001).Further subgroup analysis showed that serum total cholesterol and low density lipoprotein cholesterol lev-els were significantly increased in patients with high-level PCSK9,while patients with high-level MIF had higher inflamma-tory coefficients such as systemic inflammatory response index(SIRI)and systemic immune-inflammation index(SII)(all P＜0.05).Conclusion Serum levels of PCSK9 and MIF are positively correlated with the severity of coronary artery stenosis.High levels of serum PCSK9 and MIF are independent risk factors for CHD.

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.

Aim To explore the relationship between serum levels of proprotein convertase subtilisin/kexin type 9(PCSK9),macrophage migration inhibitory factor(MIF)and the severity of coronary artery lesions in patients with coro-nary heart disease(CHD).Methods A cross-sectional study was conducted involving 139 patients with CHD and 69 control subjects who underwent coronary angiography during the same period,all of whom were admitted to the People's Hospital of Xinjiang Uygur Autonomous Region from November 2023 to May 2024.Clinical data and coronary angiography results were collected,and the severity of coronary artery stenosis was quantitatively assessed using the Gensini score.Pa-tients with the Gensini scores＞0 were classified into three groups based on tertiles:the mild stenosis group(1～18 points,54 cases),the moderate stenosis group(19～36 points,54 cases),and the severe stenosis group(＞36 points,54 ca-ses).Serum levels of PCSK9 and MIF were measured by ELISA kit.Results Serum levels of PCSK9 and MIF were significantly higher in the CHD group than those in the control group(P＜0.05).Multivariable Logistic regression analy-sis revealed that high levels of serum PCSK9 and MIF were independent risk factors for CHD.Spearman correlation analy-sis showed that serum PCSK9 and MIF levels were positively correlated with Gensini score(rs=0.619 6 and r,=0.411 4,both P＜0.001).Further subgroup analysis showed that serum total cholesterol and low density lipoprotein cholesterol lev-els were significantly increased in patients with high-level PCSK9,while patients with high-level MIF had higher inflamma-tory coefficients such as systemic inflammatory response index(SIRI)and systemic immune-inflammation index(SII)(all P＜0.05).Conclusion Serum levels of PCSK9 and MIF are positively correlated with the severity of coronary artery stenosis.High levels of serum PCSK9 and MIF are independent risk factors for CHD.

Humans ; Glycated Hemoglobin ; Diabetes Mellitus, Type 1 ; Blood Glucose ; Diabetes Mellitus, Type 2

Abstract The COVID-19 pandemic has posed a series of complex challenges. COVID-19 in children and adolescents is generally less severe than in adults and the elderly; however, some children and adolescents may experience severe complications and adverse health effects even after mild or asymptomatic COVID-19 infections. The article focuses on gathering the epidemic characteristics, health impact, risk factors, prevention and control measures, and vaccination status of children and adolescents with COVID-19 infection to provide recommendations for protecting children and adolescents in the post COVID-19 era.

Objectives:To investigate the effect of inhibition of long non-coding RNA(lnc RNA)in human metastasis associated lung adenocarcinoma transcript 1(MALAT1)on glycolipitoxicity-induced human umbilical vein endothelial cell dysfunction. Methods:Human umbilical vein endothelial cells were treated with glucose and palmitic acid in vitro to establish the glycolipitoxic endothelial cell models.Following groups were examined:control group,high-glucose and high-fat group,high-glucose and high-fat + non-targeting RAN control group,high-glucose and high-lipid+MALAT1 siRNA group,and high-glucose and high-lipid+MAPK1 siRNA group.RT-qPCR was used to detect the mRNA expression of MALAT1 and MAPK1.Western blot was used to detect the expression levels of autophagy,mitochondrial fusion division,apoptosis,and pathway-related proteins.Immunofluorescence confocal localization was used to detect the fluorescence colocalization of autophagy and lysosome-related proteins.The number of autophagolysosomes in endothelial cells was observed by transmission electron microscopy.Mitochondrial probe staining was used to detect mitochondrial morphology,immunofluorescence was used to detect intracellular reactive oxygen species(ROS)production,flow cytometry was used to detect the apoptosis of cells in each group,cell proliferation and scratch assays were used to detect the proliferation and migration ability of cells in different groups at different time points.The angiogenesis was quantified by counting the number of new blood vessels in each group. Results:Compared with the control group,the expression of lncRNA MALAT1 mRNA and the expression of phosphorylated mito-activated protein kinase 1(p-MAPK1)were upregulated(both P＜0.05)and the expression of phosphorylated mammalian target protein(p-mTOR)was downregulated in the high-glucose and high-fat group and the high-sugar and high-fat control group(all P＜0.01).Compared with the high-glucose and high-fat non-targeting RNA control group,the expressions of microtubule-associated protein 1A/1B-light chain 3(LC3)and p62 were downregulated(P＜0.01,P＜0.05),LC3 and lysosome-associated membrane protein 2(LAMP2)protein co-localized positive fluorescence particles were increased(both P＜0.01),number of lysosomes were decreased,the expression of ROS was decreased(P＜0.01),the expression level of mitochondrial fusion protein optic nerve atrophin 1(OPA1)was increased(P＜0.05),the expressions of cleaved caspase-3 and BCL-2-related X protein(BAX)were decreased and BCL-2 was increased(all P＜0.05),cell proliferation,migration,and tube-forming ability were increased(all P＜0.01),and the expression of p-MAPK1 was decreased(P＜0.05)and p-mTOR expression was increased(both P＜0.05)in the high-glucose and high-lipid+si-MALAT1 group.Compared with the high-glucose and high-fat non-targeting RNA control group,the expression of p-MAPK1 in endothelial cells was decreased and the expression of p-mTOR was increased in the high-glucose and high-lipid+si-MAPK1 group(both P＜0.01). Conclusions:Inhibition of lncRNA MALAT1 expression can reduce the level of mitophagy in glycolipidotoxic environments,reduce apoptosis of endothelial cells and improve endothelial cell function,which may be related to the regulation of MAPK1/mTOR signaling pathway.

Objective:To analyze the effect of traditional Chinese medicine fasting therapy on atherogenic index of plasma and metabolic indices in patients with metabolic syndrome.Methods:A case-control study was conducted on 90 patients with metabolic syndrome who were treated at Wenzhou Traditional Chinese Medicine Hospital between June 2021 and June 2023. The patients were divided into two groups with 45 patients in each group using a random number table method. The control group was given a normal diet, while the observation group underwent traditional Chinese medicine fasting therapy. Blood lipid indexes, atherogenic index of plasma, Homeostasis Model Assessment of Insulin Resistance index, anthropometric indices, and oxidative stress response were compared between the two groups.Results:After treatment, the levels of low-density lipoprotein cholesterol, total cholesterol, and triglyceride in the observation group were (2.11 ± 0.26) mmol/L, (4.31 ± 0.26) mmol/L, and (1.39 ± 0.26) mmol/L, respectively. They were significantly lower than those in the control group [(2.95 ± 0.34) mmol/L, (5.24 ± 0.33) mmol/L, (2.68 ± 0.41) mmol/L, t = 13.16, 14.85, 17.82, all P < 0.05]. After treatment, the level of high-density lipoprotein cholesterol was significantly higher in the observation group than in the control group [(1.18 ± 0.09) mmol/L vs. (1.03 ± 0.04) mmol/L, t = 10.21, P < 0.001]. After treatment, atherogenic index of plasma and Homeostasis Model Assessment of Insulin Resistance index in the observation group were (0.10 ± 0.04) and (5.12 ± 0.42), respectively in the observation group. They were significantly lower than those in the control group [(0.28 ± 0.10), (5.80 ± 0.67), t = 11.21, 5.76, both P < 0.001]. After treatment, waist circumference, hip circumference, and visceral adiposity index in the observation group were (91.05 ± 4.26) cm, (98.16 ± 5.06) cm, and (3.94 ± 0.52), respectively. They were significantly lower than those in the control group [(95.55 ± 9.87) cm, (102.64 ± 9.84) cm, (5.66 ± 1.04), t = 2.80, 2.71, 9.92, all P < 0.05). After treatment, the level of superoxide dismutase in the observation group was significantly higher than that in the control group [(101.52 ± 13.52) U/mL vs. (80.01 ± 6.82) U/mL, t = 9.52, P < 0.001]. After treatment, the level of malondialdehyde in the observation group was significantly lower than that in the control group [(4.41±0.35) nmol/L vs. (6.26 ± 0.61) nmol/L, t = 17.64, P < 0.001). Conclusion:Traditional Chinese medicine fasting therapy can effectively reduce the atherogenic index of plasma, blood lipid level, waist circumference, hip circumference, and visceral adiposity index in patients with metabolic syndrome. It also reduces oxidative stress reactions and is highly effective.