Objective:To develop outcome indicators for the implementation of comprehensive inter-ventions targeting the multimorbidity of myopia and obesity in children and adolescents,providing a basis for the co-prevention of multimorbidity and the outcome measurement of implementation research in children and adolescents.Methods:Based on the RE-AIM framework,a preliminary set of indicators was constructed.The Delphi method was employed,with experts scoring and providing feedback on the proposed indicators via questionnaires.After each round of consultation,expert enthusiasm index,authority coefficient,coordination degree,and consensus level were calculated.Expert opinions were col-lected and analyzed to modify,delete,or add indicators based on consultation results and screening crite-ria.Two Delphi rounds were conducted until consensus was achieved.Results:A total of 28 experts par-ticipated actually in both rounds.The Kendall's W coefficients for the two rounds of expert consultation were0.352(x2=413.952,P＜0.001)and 0.499(x2=405.044,P＜0.001),both statistically sig-nificant.The final outcome indicators for implementation research on comprehensive interventions for myopia and obesity comorbidity in children and adolescents included five primary dimensions with 13 secondary and 20 tertiary indicators.The dimension of reach included the number of children and adoles-cents involved,participant representativeness,and full-course participation representativeness.The di-mension of effectiveness included multimorbidity incidence,myopia incidence,spherical equivalent,body mass index(BMI),overweight and obesity prevalence,waist-to-height ratio,comprehensive health knowledge score,and comprehensive health behavior score.The dimension of adoption covered school representativeness and representativeness of school nurses and teachers involved in implementation.The dimension of implementation included fidelity,content modification,and cost.The dimension of mainte-nance included individual health outcomes and organizational sustainment.Conclusion:This study developed implementation outcome indicators for comprehensive interventions targeting multimorbidity of myopia and obesity among the children and adolescents based on the RE-AIM framework.These indica-tors can serve as a reference for optimizing intervention research strategies related to common multimor-bidity among children and adolescents in China.

This study was aimed at preliminarily investigating the molecular biological functions of the PPE65 protein from Myco-bacterium tuberculosis,and providing foundational data for tuberculosis prevention and control.The basic biological properties of the PPE65 gene-encoded protein were predicted with bioinformatics tools.Sequence information for the Mycobacterium tuberculosis Rv3621c gene and PPE65 protein was retrieved from the NCBI database.The Rv3621c gene was amplified through PCR with the H37Rv genome as a template,then cloned into the pET22b(+)expression vector.The recombinant pET22b(+)-PPE65 plasmid was transformed into Escherichia coli BL21(DE3)competent cells for IPTG-induced expression.Solubility analysis,purification,and identification of the recombinant PPE65 protein were performed.BEAS-2B cells were treated with various concentrations of PPE65 protein for 24 h,and cell proliferation was assessed with CCK-8 assays.PPE65 was found to be composed of 413 amino acids and to have a molecular formula of C????H????N???O???S??,a relative molecular mass of 40 679.88,a theoretical isoelectric point of 4.60,an ali-phatic index of 81.94,and an average hydrophilicity value of 0.319,thus indicating a stable hydrophobic protein lacking signal pep-tides or transmembrane domains.Secondary structure analysis revealed 53.03%α-helix(Hh),2.66%β-sheet(Ee),and 44.31%ran-dom coil(Cc).Bioinformatics predictions identified 38 B-cell epitopes and 22 CTL/Th-cell epitopes.The full-length PPE65 gene(1 308 bp)was confirmed through double restriction enzyme digestion and sequencing,thereby validating the correct construction of the pET22b(+)-PPE65 recombinant plasmid.SDS-PAGE analysis demonstrated that the recombinant protein was found in inclusion bodies,and a single band at 43.7 kDa was observed after purification.Western blotting revealed specific binding to mouse-derived His monoclonal antibodies,thereby confirming successful expression of the PPE65 protein.BEAS-2B cells treated with a PPE65 protein concentration gradient(2.5-20 μg/mL)exhibited a dose-dependent increase in cell number.Compared with those in the PBS control group,TGF-β relative expression levels were significantly higher in all treatment groups(t2.5=4.893,P<0.001,t5.0=4.640,P<0.05,t10=7.535,P<0.05,t20=16.44,P<0.000 1).This study elucidated the structural characteristics of the PPE65 protein,successfully obtained the recombinant protein through prokaryotic expression and purification,and demonstrated its ability to promote BEAS-2B cell proliferation.The underlying mechanism might involve suppression of TGF-β/S mad signaling pathway activation.These findings provide a theoretical basis for understanding the role and regulatory mechanisms of PPE65 during M.tuberculosis infection.

Objective:To analyze the application of Sengstaken-Blakemore tubes combined with endoscopic intervention on the prognosis of patients with acute esophageal and gastric variceal rupture and bleeding due to portal hypertension in liver cirrhosis, and to explore the impact of Sengstaken-Blakemore tubes combined with endoscopic intervention on the prognosis of patients.Methods:A retrospective study included patients with liver cirrhosis who visited the Zhongshan Hospital, Fudan University from January 2023 to July 2024 due to acute upper gastrointestinal bleeding, had no response to drug treatment and unstable hemodynamics, and received the Sengstaken-Blakemore tubes compression hemostasis. According to whether endoscopic intervention was received or not, the patients were divided into the endoscopic intervention group and the non-endoscopic intervention group. The medication and subsequent treatment of the patients were analyzed, and the survival of the patients 28 days after bleeding was followed up. Kaplan-Meier survival analysis and Cox regression analysis were used to evaluate the effect of endoscopic intervention on the 28-day mortality rate of patients.Results:A total of 31 cases successfully had three-lumen two-cystic tubes indwelled. Among them, 8 cases further received endoscopic examination and intervention, and 23 cases did not receive endoscopic examination. During the 28-day follow-up after bleeding, a total of 20 cases died. The Kaplan-Meier survival analysis showed that the 28-day mortality rate in the endoscopic intervention group was significantly lower than that in the non-endoscopic intervention group (25.0% vs 78.3%, P=0.007). Multivariate Cox regression analysis showed that endoscopic intervention was an independent protective factor for 28-day mortality ( HR: 0.14, 95% CI: 0.03-0.63, P=0.01). In addition, the severity of bleeding and prothrombin time were also important factors affecting the prognosis of patients. Conclusions:The Sengstaken-Blakemore tubes, as an important hemostasis method for acute esophageal and gastric variceal rupture and bleeding, can buy time for subsequent endoscopic treatment, but the risk of complications is relatively high. Creating conditions for early endoscopic intervention as early as possible can significantly reduce the mortality rate of patients and improve the prognosis of patients with liver cirrhosis.

Objective The reference interval of venous whole blood red cells and related parameters of healthy children preschool stage 3 to 6 years in Panzhihua was established to provide practical experimental basis for pediatric clinical diagnosis and treatment.Methods From January to July 2023,1 467 eligible healthy children from 3 to 6 years old for physical examination were collected as study subjects,including 762 boys and 705 girls.Red blood cell(RBC),hemoglobin(Hb),hematocrit(HCT),mean red blood cell volume(MCV),mean corpuscular hemoglobin(MCH),mean corpuscular hemoglobin concentration(MCHC)and red cell volume distribution width(RDW)were measured by Sysmex XN-1000 automatic blood analyzer.According to sex and age groups,the differences of the above indexes between different gender and age groups were compared,and the reference interval of venous whole blood red cells and related parameters of healthy children aged 3 to 6 years old in Panzhihua was established.At the same time,25 samples(including 13 boys and 12 girls)were selected to verify the newly established reference interval.Results HCT and RDW-SD reference intervals between boys and girls preschool stage 3～6 in the Panzhihua area,and the differences were not statistically significant(U=0.000,0.795,all P>0.05).The reference intervals for RBC,MCV,MCH and RDW-CV show statistically significant differences between boys and girls(U=2.829～5.753,all P＜0.05).There were gender and age differences in the reference intervals of Hb and MCHC(U=2.599,4.368,all P＜0.01).In the 3～6 years,the basal levels of RBC,HGB and HCT tended to increase gradually with age,while the overall base levels of RBC,HGB,MCHC,and RDW-CV in boys were higher than those in girls.And there were different degrees of difference with the reference range of industry standards.Also verify through the new reference interval.Conclusion With differences in RBC and related reference intervals of preschool stage children in different regions,it is necessary to establish a reference interval suitable for special population in Panzhihua.

Objective To develop the Ego Depletion Scale for Type 2 Diabetes Patients and evaluate its reliability and validity,and to provide a specific assessment tool for evaluating ego-depletion in self-management.Methods Guided by the self-control strength model,the initial scale was constructed through literature review,semi-structured interviews,2 rounds of expert consultation,and a pilot survey.A convenience sampling method was employed to recruit 460 patients with Type 2 Diabetes from the endocrinology department of a tertiary hospital in Wuhan,Hubei Province,between April and July 2024.They were randomly divided into 2 subsets for exploratory factor analysis and confirmatory factor analysis.Results A total of 451 valid questionnaires were collected.Exploratory factor analysis extracted 6 common factors,with a cumulative variance contribution of 73.231％.In confirmatory factor analysis,an item was deleted due to failing to meet the standardized loading value criterion.The revised Ego Depletion Scale for Type 2 Diabetes Patients comprised 6 dimensions and 22 items.The total Cronbach's α coefficient was 0.911;split-half reliability was 0.744;the content validity index was 0.860.Correlation coefficients between the total score and scores of each dimension of the scale and the total score of the Self-Regulatory Fatigue Scale ranged from 0.558 to 0.946(P＜0.001).Conclusion The scale exhibits robust reliability and validity,serving as a scientifically instrument for assessing ego depletion in patients with Type 2 Diabetes.

Objective:To analyze the application of Sengstaken-Blakemore tubes combined with endoscopic intervention on the prognosis of patients with acute esophageal and gastric variceal rupture and bleeding due to portal hypertension in liver cirrhosis, and to explore the impact of Sengstaken-Blakemore tubes combined with endoscopic intervention on the prognosis of patients.Methods:A retrospective study included patients with liver cirrhosis who visited the Zhongshan Hospital, Fudan University from January 2023 to July 2024 due to acute upper gastrointestinal bleeding, had no response to drug treatment and unstable hemodynamics, and received the Sengstaken-Blakemore tubes compression hemostasis. According to whether endoscopic intervention was received or not, the patients were divided into the endoscopic intervention group and the non-endoscopic intervention group. The medication and subsequent treatment of the patients were analyzed, and the survival of the patients 28 days after bleeding was followed up. Kaplan-Meier survival analysis and Cox regression analysis were used to evaluate the effect of endoscopic intervention on the 28-day mortality rate of patients.Results:A total of 31 cases successfully had three-lumen two-cystic tubes indwelled. Among them, 8 cases further received endoscopic examination and intervention, and 23 cases did not receive endoscopic examination. During the 28-day follow-up after bleeding, a total of 20 cases died. The Kaplan-Meier survival analysis showed that the 28-day mortality rate in the endoscopic intervention group was significantly lower than that in the non-endoscopic intervention group (25.0% vs 78.3%, P=0.007). Multivariate Cox regression analysis showed that endoscopic intervention was an independent protective factor for 28-day mortality ( HR: 0.14, 95% CI: 0.03-0.63, P=0.01). In addition, the severity of bleeding and prothrombin time were also important factors affecting the prognosis of patients. Conclusions:The Sengstaken-Blakemore tubes, as an important hemostasis method for acute esophageal and gastric variceal rupture and bleeding, can buy time for subsequent endoscopic treatment, but the risk of complications is relatively high. Creating conditions for early endoscopic intervention as early as possible can significantly reduce the mortality rate of patients and improve the prognosis of patients with liver cirrhosis.

Objective To elucidate the molecular mechanism of shugan formula in regulating intestinal flora in liver cancer using network pharmacology.Methods The active components and targets of shugan formula were screened using the TCMSP and BATMAN-TCM databases.Targets associated with liver cancer and intestinal flora were extracted from the GeneCards and OMIM databases.The in-tersection targets between the drug and disease were identified using a Venn diagram.Kyoto Encyclopedia of Genes and Genomes(KEGG)and Gene Ontology(GO)enrichment analysis were performed on the intersection targets.Molecular docking technology was em-ployed to validate the binding activities of key components,such as quercetin,kaempferol,with core targets,including TP53 and STAT3.The correlation between targets and the prognosis of liver cancer was assessed through survival analysis.Results KEGG enrichment anal-ysis indicated that shugan formula was primarily enriched in pathways such as cancer pathways.GO enrichment analysis suggested its in-volvement in regulating processes like apoptosis.Molecular docking demonstrated stable binding between key components and core targets.Survival analysis revealed that high expression of tumor protein p53gene(TP53)shortened the overall survival of liver cancer patients,whereas high expression of signal transducer and activator of transcription 3(STAT3)prolonged survival.Conclusion Shugan formula may upregulate STAT3 expression,inhibit TP53 expression,modulate relevant signaling pathways,improve the structure of intestinal flora in liver cancer,and thereby inhibit the proliferation and metastasis of liver cancer cells.

Objective To elucidate the molecular mechanism of shugan formula in regulating intestinal flora in liver cancer using network pharmacology.Methods The active components and targets of shugan formula were screened using the TCMSP and BATMAN-TCM databases.Targets associated with liver cancer and intestinal flora were extracted from the GeneCards and OMIM databases.The in-tersection targets between the drug and disease were identified using a Venn diagram.Kyoto Encyclopedia of Genes and Genomes(KEGG)and Gene Ontology(GO)enrichment analysis were performed on the intersection targets.Molecular docking technology was em-ployed to validate the binding activities of key components,such as quercetin,kaempferol,with core targets,including TP53 and STAT3.The correlation between targets and the prognosis of liver cancer was assessed through survival analysis.Results KEGG enrichment anal-ysis indicated that shugan formula was primarily enriched in pathways such as cancer pathways.GO enrichment analysis suggested its in-volvement in regulating processes like apoptosis.Molecular docking demonstrated stable binding between key components and core targets.Survival analysis revealed that high expression of tumor protein p53gene(TP53)shortened the overall survival of liver cancer patients,whereas high expression of signal transducer and activator of transcription 3(STAT3)prolonged survival.Conclusion Shugan formula may upregulate STAT3 expression,inhibit TP53 expression,modulate relevant signaling pathways,improve the structure of intestinal flora in liver cancer,and thereby inhibit the proliferation and metastasis of liver cancer cells.

This study was aimed at preliminarily investigating the molecular biological functions of the PPE65 protein from Myco-bacterium tuberculosis,and providing foundational data for tuberculosis prevention and control.The basic biological properties of the PPE65 gene-encoded protein were predicted with bioinformatics tools.Sequence information for the Mycobacterium tuberculosis Rv3621c gene and PPE65 protein was retrieved from the NCBI database.The Rv3621c gene was amplified through PCR with the H37Rv genome as a template,then cloned into the pET22b(+)expression vector.The recombinant pET22b(+)-PPE65 plasmid was transformed into Escherichia coli BL21(DE3)competent cells for IPTG-induced expression.Solubility analysis,purification,and identification of the recombinant PPE65 protein were performed.BEAS-2B cells were treated with various concentrations of PPE65 protein for 24 h,and cell proliferation was assessed with CCK-8 assays.PPE65 was found to be composed of 413 amino acids and to have a molecular formula of C????H????N???O???S??,a relative molecular mass of 40 679.88,a theoretical isoelectric point of 4.60,an ali-phatic index of 81.94,and an average hydrophilicity value of 0.319,thus indicating a stable hydrophobic protein lacking signal pep-tides or transmembrane domains.Secondary structure analysis revealed 53.03%α-helix(Hh),2.66%β-sheet(Ee),and 44.31%ran-dom coil(Cc).Bioinformatics predictions identified 38 B-cell epitopes and 22 CTL/Th-cell epitopes.The full-length PPE65 gene(1 308 bp)was confirmed through double restriction enzyme digestion and sequencing,thereby validating the correct construction of the pET22b(+)-PPE65 recombinant plasmid.SDS-PAGE analysis demonstrated that the recombinant protein was found in inclusion bodies,and a single band at 43.7 kDa was observed after purification.Western blotting revealed specific binding to mouse-derived His monoclonal antibodies,thereby confirming successful expression of the PPE65 protein.BEAS-2B cells treated with a PPE65 protein concentration gradient(2.5-20 μg/mL)exhibited a dose-dependent increase in cell number.Compared with those in the PBS control group,TGF-β relative expression levels were significantly higher in all treatment groups(t2.5=4.893,P<0.001,t5.0=4.640,P<0.05,t10=7.535,P<0.05,t20=16.44,P<0.000 1).This study elucidated the structural characteristics of the PPE65 protein,successfully obtained the recombinant protein through prokaryotic expression and purification,and demonstrated its ability to promote BEAS-2B cell proliferation.The underlying mechanism might involve suppression of TGF-β/S mad signaling pathway activation.These findings provide a theoretical basis for understanding the role and regulatory mechanisms of PPE65 during M.tuberculosis infection.

Objective The reference interval of venous whole blood red cells and related parameters of healthy children preschool stage 3 to 6 years in Panzhihua was established to provide practical experimental basis for pediatric clinical diagnosis and treatment.Methods From January to July 2023,1 467 eligible healthy children from 3 to 6 years old for physical examination were collected as study subjects,including 762 boys and 705 girls.Red blood cell(RBC),hemoglobin(Hb),hematocrit(HCT),mean red blood cell volume(MCV),mean corpuscular hemoglobin(MCH),mean corpuscular hemoglobin concentration(MCHC)and red cell volume distribution width(RDW)were measured by Sysmex XN-1000 automatic blood analyzer.According to sex and age groups,the differences of the above indexes between different gender and age groups were compared,and the reference interval of venous whole blood red cells and related parameters of healthy children aged 3 to 6 years old in Panzhihua was established.At the same time,25 samples(including 13 boys and 12 girls)were selected to verify the newly established reference interval.Results HCT and RDW-SD reference intervals between boys and girls preschool stage 3～6 in the Panzhihua area,and the differences were not statistically significant(U=0.000,0.795,all P>0.05).The reference intervals for RBC,MCV,MCH and RDW-CV show statistically significant differences between boys and girls(U=2.829～5.753,all P＜0.05).There were gender and age differences in the reference intervals of Hb and MCHC(U=2.599,4.368,all P＜0.01).In the 3～6 years,the basal levels of RBC,HGB and HCT tended to increase gradually with age,while the overall base levels of RBC,HGB,MCHC,and RDW-CV in boys were higher than those in girls.And there were different degrees of difference with the reference range of industry standards.Also verify through the new reference interval.Conclusion With differences in RBC and related reference intervals of preschool stage children in different regions,it is necessary to establish a reference interval suitable for special population in Panzhihua.