Purpose: Due to the preservation of the entire stomach after endoscopic resection, the occurrence of metachronous gastric cancer (MGC) remains a possibility. In this study, we investigated the incidence and risk factors for MGC in patients with early gastric cancer who underwent endoscopic submucosal dissection (ESD) and successfully eradicatedHelicobacter pylori. Materials and Methods: A retrospective analysis was conducted of 1,191 patients who underwent ESD and successfully eradicated H. pylori at the Affiliated Drum Tower Hospital of Nanjing University. Endoscopic surveillance was performed at 3, 6, and 12 months post-resection, and annually thereafter. MGC was defined as the development of a new cancer at a site other than the primary gastric cancer site, at least 1 year after the initial endoscopic resection. Results: A total of 77 patients were diagnosed with MGC during a median follow-up of 41.5 months. Kaplan-Meier analysis showed a 5-year cumulative incidence of MGC of 9.4% after successful H. pylori eradication. Multivariate analysis of the training set using Cox proportional hazards models identified male sex, severe atrophic gastritis, multiple gastric cancers before H. pylori eradication, and smoking history as independent risk factors for MGC.The nomogram exhibited favorable discrimination, with area under the curves of 0.767 and 0.822 in the training set and 0.724 and 0.745 in the testing set at 3 and 5 years, respectively. Conclusions Patients with gastric cancer who undergo endoscopic resection, even after successful H. pylori eradication, should undergo annual and continuous endoscopic surveillance for MGC.

Objective To explore the application value of combining the ultrasound breast imaging reporting and data system(BI-RADS)classification with serum fibroblast growth factor receptor 1(FGFR1)and growth differentiation factor 3(GDF3)in the differential diagnosis of benign and malignant breast masses.Methods A total of 159 patients with breast masses were selected and divided into the benign mass group(n=83)and the malignant mass group(n=76)based on postoperative pathological diagnosis.All patients underwent ultrasound examination,and enzyme-linked immunosorbent assay(ELISA)was applied to detect serum levels of FGFR1 and GDF3.Receiver operating characteristic(ROC)curve was applied to analyze the diagnostic value of ultrasound BI-RADS classification and serum FGFR1 and GDF3 levels for benign and malignant breast masses.Kappa test was applied to analyze the consistency between various diagnostic methods and pathological diagnosis.Results The serum levels of FGFR1 and GDF3,the proportions of irregular morphology,unclear boundaries,spiculation,microcalcifications,blood flow grade Ⅱ-Ⅲ and posterior echo attenuation,RI and PI were higher in the malignant tumor group than those in the benign tumor group(P＜0.05).The area under the curve(AUC)of FGFR1,GDF3 and ultrasound BI-RADS classification in the differential diagnosis of benign and malignant breast masses separately and in combination was 0.802(95%CI:0.732-0.871),0.817(95%CI:0.751-0.884),0.848(95%CI:0.784-0.912)and 0.956(95%CI:0.918-0.993),respectively.The combined diagnosis was more effective than that of the individual diagnosis of each indicator.The consistency between the individual and combined diagnosis of benign and malignant breast masses and pathological diagnosis showed that the Kappa values were 0.517,0.514,0.688 and 0.912,respectively,with the highest consistency observed in the combined diagnosis(P＜0.05).Conclusion Ultrasound BI-RADS classification combined with serum FGFR1 and GDF3 has high application value in the differential diagnosis of benign and malignant breast masses.

Objective To explore the predictive value of coronary artery calcium score(CACS)combined with C-X-C motif chemokine ligand 12(CXCL12)and soluble CD40 ligand(sCD40L)for major adverse cardiovascular events(MACE)within 5 years in patients with stable coronary artery disease(SCAD).Methods A total of 206 elderly SCAD patients admitted in our depart-ment from March 2015 to November 2018 were recruited,and according to whether MACE occurred within 5 years of follow-up,they were divided into MACE group(42 cases)and control group(164 cases).Multivariate logistic regression analysis was performed to identify the inde-pendent predictors of MACE.ROC curve analysis was applied to evaluate the predictive value of the combined model,and its area under curve(AUC)value was calculated.Results Compared with the control group,the MACE group had significantly older age,larger proportions of hyper-tension and diabetes mellitus,higher CACS,and elevated CXCL12 and sCD40L levels(P＜0.05,P＜0.01).Multivariate logistic regression analysis revealed that age,hypertension,diabetes melli-tus,CACS,CXCL12 and sCD40L were independent risk factors for MACE in the elderly SCAD patients within 5 years(P＜0.05,P＜0.01).ROC curve analysis indicated that the AUC value of CACS,CXCL12,sCD40L,and these three indicators combined together in predicting MACE in elderly SCAD patients within 5 years was 0.872(95％CI:0.819-0.915),0.768(95％CI:0.704-0.824),0.726(95％CI:0.660-0.786),and 0.935(95％CI:0.893-0.965),respectively,with the value of the combination obviously higher than that of each indicator alone(P＜0.01).Conclusion Increased CACS,CXCL12 and sCD40L have predictive value for MACE in elderly SCAD patients,and the combination of the three indicators has obvious advantages in the prediction.

Objective To investigate the role of cathepsin K(CTK)and secreted frizzled-related protein 2(sFRP2)in ventricular remodeling in very old patients with acute myocardial infarction(AMI)after percutaneous coronary intervention(PCI),and analyze the correlation.Methods A total of 192 very old AMI patients undergoing PCI in our department from January 2020 to December 2023 were recruited,and based on clinical outcomes at 6 months after surgery,they were assigned into a ventricular remodeling group(74 cases)and a non-ventricular remodeling group(118 cases).The clinical data and postoperative biochemical indicators were collected in the two groups.Multivariate logistic regression analysis was performed to assess the correlation between CTK and sFRP2 levels and ventricular remodeling.ROC curves were plotted to analyze the predictive value of CTK and sFRP2 in ventricular remodeling.Results The ventricular remodeling group had significantly longer length of hospital stay,larger left ventricular end-diastolic diameter,and higher levels of CTK,sFRP2,C-reactive protein(CRP)and IL-6,but lower left ventricular ejection fraction than the non-ventricular remodeling group(P＜0.05,P＜0.01).Multivariate logistic regression analysis suggested that CTK and sFRP2 were independent risk factors for postoperative ventricular remodeling in very old AMI patients(P＜0.01).ROC curve analysis manifested that the area under curve value of CTK,sFRP2,CRP and IL-6 combined together in predicting ventricular remodeling in very old AMI patients after PCI was 0.892,with a sensitivity of 79.73％and a specificity of 88.14％(P＜0.01).Conclusion CTK and sFRP2 are significantly increased in ventricular remodeling in very old AMI patients after PCI,and are closely related to the occurrence of ventricular remodeling.They can be used as potential biomark-ers in the prediction of ventricular remodeling and provide important reference for clinical diagno-sis and treatment.

Objective:To explore the intervention effects of the skill training for parents with autism child (STPAC) on toddlers with autism spectrum disorder (ASD).Methods:A multicenter non-randomized concurrent controlled study design was conducted. Thirty children with ASD aged 15-30 months, first diagnosed at the Children′s Hospital of Fudan University, the First Hospital of Jilin University, and Chengdu Women′s and Children′s Central Hospital from 2019 to 2020, were enrolled in the STPAC group. Thirty children with ASD who visited the same hospitals during the same period but refused the STPAC intervention were selected as the control group. The STPAC group received an 8-week intervention (3 h/week) followed by quarterly follow-ups for 1 year, while the control group voluntarily chose community-based routine interventions. The Griffiths development scales-Chinese (GDS-C) was used to assess the developmental levels, and the communication and symbolic behavior scales developmental profile infant-toddler checklist (CSBS-DP-ITC) was completed by the primary caregivers to evaluate social, language and symbolic behavior. The independent samples t-tests or Mann-Whitney U tests, etc.was used for inter-group comparison. The paired t-tests or Wilcoxon signed-rank tests, etc. was used for inter-group pre-post intervention comparison. Results:The STPAC group included 30 children (22 males and 8 females, aged (23.9±2.2) months), and the control group included 30 children (20 males and 10 females, aged (24.2±2.6) months). Before the intervention, there were no statistically differences in GDS-C development quotient (DQ) and CSBS-DP-ITC scores between groups (all P>0.05). After 1-year intervention, GDS-C DQ in personal-social, hearing-language, hand-eye coordination, performance domains of STPAC group and GDS-C DQ in personal-social, hearing-language domains of control group were all increased (all P<0.01). After 1-year intervention, CSBS-DP-ITC scores of both groups were all improved in socia, language, symbolic behavior, and total scores (all P<0.001). GDS-C DQ changes before and after 1 year of intervention in hearing-language, hand-eye coordination, performance domains of the STPAC group were all higher the those of control group (34(15, 48 vs. 10(-4, 39), 11±20 vs. -1±19, 23±25 vs. 8±22, all P<0.05). CSBS-DP-ITC scores changes before and after 1 year of intervention in social and total scores of the STPAC group were both higher the those of control group (10(5, 30) vs. 3(1, 7), 26±17 vs. 11±8, both P<0.001). Conclusion:Compared with the community routine interventions, the STPAC better improves the language, hand-eye coordination, visual-spatial, social communication, and play skills in ASD toddlers.

OBJECTIVE: EPF3 is a fibrinolysin monomer isolated and purified from Pheretima vulgaris Chen, an earthworm used in traditional Chinese medicine as Dilong for treating blood stasis syndrome. Its composition, anticoagulant and fibrinolytic activities, and relevant mechanisms have been confirmed through in vitro experiments. However, whether it has antithrombotic effects in vivo and can be absorbed by the gastrointestinal tract is unknown. This study evaluates the antithrombotic effect in zebrafish and investigates the gastrointestinal stability and intestinal absorption mechanism of this protein in vitro. METHODS: The antithrombotic effect of EPF3 in vivo was verified using the zebrafish thrombus model induced by arachidonic acid and FeCl₃. Then, the protein bands of EPF3 incubated with simulated gastric fluid (SGF), simulated intestinal fluid (SIF), and homogenate of Caco-2 cells (HC2C) were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to evaluate its gastrointestinal stability. Finally, the transport behavior and absorption mechanism of EPF3 were studied using Caco-2 cell monolayer. RESULTS: EPF3 could significantly enhance the returned blood volume and blood flow velocity in zebrafish with platelet aggregation thrombus induced by arachidonic acid. It could also prolong the formation time of tail artery thrombus and increase the blood flow velocity in zebrafish with vessel injury thrombus induced by FeCl₃. EPF3 was stable in SIF and HC2C and unstable in SGF. The permeability of EPF3 in Caco-2 monolayer was time-dependent and concentration-dependent. The efflux ratio was less than 1.2 during transport, and the transport behavior was not affected by inhibitors. EPF3 could reversibly reduce the expression of tight junction-related proteins, including zonula occludens-1, occludin, and claudin-1 in Caco-2 cells. CONCLUSION EPF3 could play a thrombolytic and antithrombotic role in zebrafish. It could be transported and absorbed into the intestine through cellular bypass pathway by opening the intestinal epithelium tight junction. This study provides a scientific explanation for the antithrombotic effect of earthworm and provides a basis for the feasibility of subsequent development of EPF3 as an antithrombotic enteric-soluble preparation. Please cite this article as: Zhong WL, Yang JQ, Liu H, Wu YL, Shen HJ, Li PY, Du SY. Antithrombotic effect in zebrafish of a fibrinolytic protein EPF3 from Dilong (Pheretima vulgaris Chen) and its transport mechanism in Caco-2 monolayer through cell bypass pathway. J Integr Med. 2025; 23(4): 415-428.
Animals ; Zebrafish ; Humans ; Caco-2 Cells ; Fibrinolytic Agents/pharmacology* ; Thrombosis/drug therapy* ; Intestinal Absorption

Objective To explore the predictive value of coronary artery calcium score(CACS)combined with C-X-C motif chemokine ligand 12(CXCL12)and soluble CD40 ligand(sCD40L)for major adverse cardiovascular events(MACE)within 5 years in patients with stable coronary artery disease(SCAD).Methods A total of 206 elderly SCAD patients admitted in our depart-ment from March 2015 to November 2018 were recruited,and according to whether MACE occurred within 5 years of follow-up,they were divided into MACE group(42 cases)and control group(164 cases).Multivariate logistic regression analysis was performed to identify the inde-pendent predictors of MACE.ROC curve analysis was applied to evaluate the predictive value of the combined model,and its area under curve(AUC)value was calculated.Results Compared with the control group,the MACE group had significantly older age,larger proportions of hyper-tension and diabetes mellitus,higher CACS,and elevated CXCL12 and sCD40L levels(P＜0.05,P＜0.01).Multivariate logistic regression analysis revealed that age,hypertension,diabetes melli-tus,CACS,CXCL12 and sCD40L were independent risk factors for MACE in the elderly SCAD patients within 5 years(P＜0.05,P＜0.01).ROC curve analysis indicated that the AUC value of CACS,CXCL12,sCD40L,and these three indicators combined together in predicting MACE in elderly SCAD patients within 5 years was 0.872(95％CI:0.819-0.915),0.768(95％CI:0.704-0.824),0.726(95％CI:0.660-0.786),and 0.935(95％CI:0.893-0.965),respectively,with the value of the combination obviously higher than that of each indicator alone(P＜0.01).Conclusion Increased CACS,CXCL12 and sCD40L have predictive value for MACE in elderly SCAD patients,and the combination of the three indicators has obvious advantages in the prediction.

Objective To investigate the role of cathepsin K(CTK)and secreted frizzled-related protein 2(sFRP2)in ventricular remodeling in very old patients with acute myocardial infarction(AMI)after percutaneous coronary intervention(PCI),and analyze the correlation.Methods A total of 192 very old AMI patients undergoing PCI in our department from January 2020 to December 2023 were recruited,and based on clinical outcomes at 6 months after surgery,they were assigned into a ventricular remodeling group(74 cases)and a non-ventricular remodeling group(118 cases).The clinical data and postoperative biochemical indicators were collected in the two groups.Multivariate logistic regression analysis was performed to assess the correlation between CTK and sFRP2 levels and ventricular remodeling.ROC curves were plotted to analyze the predictive value of CTK and sFRP2 in ventricular remodeling.Results The ventricular remodeling group had significantly longer length of hospital stay,larger left ventricular end-diastolic diameter,and higher levels of CTK,sFRP2,C-reactive protein(CRP)and IL-6,but lower left ventricular ejection fraction than the non-ventricular remodeling group(P＜0.05,P＜0.01).Multivariate logistic regression analysis suggested that CTK and sFRP2 were independent risk factors for postoperative ventricular remodeling in very old AMI patients(P＜0.01).ROC curve analysis manifested that the area under curve value of CTK,sFRP2,CRP and IL-6 combined together in predicting ventricular remodeling in very old AMI patients after PCI was 0.892,with a sensitivity of 79.73％and a specificity of 88.14％(P＜0.01).Conclusion CTK and sFRP2 are significantly increased in ventricular remodeling in very old AMI patients after PCI,and are closely related to the occurrence of ventricular remodeling.They can be used as potential biomark-ers in the prediction of ventricular remodeling and provide important reference for clinical diagno-sis and treatment.

Objective:To explore the intervention effects of the skill training for parents with autism child (STPAC) on toddlers with autism spectrum disorder (ASD).Methods:A multicenter non-randomized concurrent controlled study design was conducted. Thirty children with ASD aged 15-30 months, first diagnosed at the Children′s Hospital of Fudan University, the First Hospital of Jilin University, and Chengdu Women′s and Children′s Central Hospital from 2019 to 2020, were enrolled in the STPAC group. Thirty children with ASD who visited the same hospitals during the same period but refused the STPAC intervention were selected as the control group. The STPAC group received an 8-week intervention (3 h/week) followed by quarterly follow-ups for 1 year, while the control group voluntarily chose community-based routine interventions. The Griffiths development scales-Chinese (GDS-C) was used to assess the developmental levels, and the communication and symbolic behavior scales developmental profile infant-toddler checklist (CSBS-DP-ITC) was completed by the primary caregivers to evaluate social, language and symbolic behavior. The independent samples t-tests or Mann-Whitney U tests, etc.was used for inter-group comparison. The paired t-tests or Wilcoxon signed-rank tests, etc. was used for inter-group pre-post intervention comparison. Results:The STPAC group included 30 children (22 males and 8 females, aged (23.9±2.2) months), and the control group included 30 children (20 males and 10 females, aged (24.2±2.6) months). Before the intervention, there were no statistically differences in GDS-C development quotient (DQ) and CSBS-DP-ITC scores between groups (all P>0.05). After 1-year intervention, GDS-C DQ in personal-social, hearing-language, hand-eye coordination, performance domains of STPAC group and GDS-C DQ in personal-social, hearing-language domains of control group were all increased (all P<0.01). After 1-year intervention, CSBS-DP-ITC scores of both groups were all improved in socia, language, symbolic behavior, and total scores (all P<0.001). GDS-C DQ changes before and after 1 year of intervention in hearing-language, hand-eye coordination, performance domains of the STPAC group were all higher the those of control group (34(15, 48 vs. 10(-4, 39), 11±20 vs. -1±19, 23±25 vs. 8±22, all P<0.05). CSBS-DP-ITC scores changes before and after 1 year of intervention in social and total scores of the STPAC group were both higher the those of control group (10(5, 30) vs. 3(1, 7), 26±17 vs. 11±8, both P<0.001). Conclusion:Compared with the community routine interventions, the STPAC better improves the language, hand-eye coordination, visual-spatial, social communication, and play skills in ASD toddlers.

Objective To investigate gender differences and age-related changes in body composition(BC)among Miao adults in Rongshui,Guangxi Province,and to provide the basis for assessing nutritional status and health.Methods With informed consent,630 Miao adults(218 males,412 females)were randomly selected for this study.Body composition was assessed using bioelectrical impedance analysis(BIA).Results Weight,fat-free mass,muscle mass,trunk muscle mass,limb muscle mass,waist-to-hip ratio(WHR),body water,presumtion of bone mass and protein were significantly higher in males than in females.And the fat mass,trunk fat mass,limb fat mass,visceral fat content,subcutaneous fat content and percentage of body fat were significantly higher in females than in males.According to the evaluation of body mass index(BMI)and WHR,the proportion of overweight and obesity of Miao adults was higher than the average level of Miao residents,and their obesity was characterized by central obesity.With age,weight,fat mass,muscle mass,fat-free mass,limb muscle mass,limb fat mass,subcutaneous fat content,percentage of body fat,body water,presumtion of bone mass,and protein of Rongshui Miao adults showed a gradual decreasing trend,while visceral fat content and WHR increased progressively.BMI in male Miao adults,along with BMI,fat mass,trunk fat mass,subcutaneous fat content,percentage of body fat,and body water in female Miao adults,showed a trend of increasing followed by decreasing,peaking at the age of 40-49 years.Conclusion The body composition of Miao adults in Rongshui,Guangxi,exhibits significant gender differences and age-related variation change patterns,which may increase the risk of sarcopenia and metabolic diseases with aging.