To construct a recombinant fowl adenovirus 4(FAdV-4)expressing the Cap protein of goose astrovirus genotype 2(GoAstV-2),the expression cassette of Cap gene was inserted into the natural 1 966 bp deletion region of the FAdV-4 genome in the infectious clone p15A-cm-FAdV4-HNJZ.The resulted recombinant plasmid p15A-cm-FAdV4-HNJZ-Cap/GoAstV-2 was linearized with restriction enzyme and transfected into chicken hepatoma cell line(LMH)to rescue the recombinant FAdV-4 expressing the Cap protein of GoAstV-2,rF Ad V4-Cap/GoAstV-2.After 15 passages in LMH cells,the recombinant rFAdV4-Cap/GoAstV-2 was identified by PCR using primers flanking the insertion site of the Cap gene expression cassette and using viral genome DNA extracted from rFAdV4-Cap/GoAstV-2 infected LMH cells as template.LMH cells were in-fected with 15th passage rFAdV4-Cap/GoAstV-2 and indirect immunofluorescence was performed with a polyclonal antibody against Cap protein as the primary antibody.Western blot was carried out with lysates of rFAdV4-Cap/GoAstV-2 infected LMH cells.The in vitro replication dynamic of the 15th passage of the rFAdV4-Cap/GoAstV-2 was also investigated in LMH cells.The results demonstrated that the Cap gene of GoAstV-2 was presented in the genome of the recombinant vi-rus rF AdV4-Cap/Go Ast V-2,and could be expressed stably.The prepared recombinant virus in this study will lay a foundation for developing inactivated bivalent vaccine candidate against co-in-fection of FAdV-4 and GoAstV-2 in goose.

Objective This study aimed to investigate the antimicrobial resistance profiles of bacterial strains isolated from pediatric intensive care units(PICU)in China for better antimicrobial therapy.Methods Clinical isolates were collected from 17 institutions,including tertiary care children's hospitals and pediatric department of tertiary general hospitals in China from January 1,2020 to December 31,2022.Antimicrobial susceptibility testing was carried out according to a unified protocol using Kirby-Bauer method or automated systems.Results were interpreted according to the breakpoints released by the Clinical and Laboratory Standards Institute(CLSI)in 2020.Results A total of 10 688 isolates were collected,including gram-positive organisms(39.2％)and gram-negative organisms(60.8％).The top three organisms were S.aureus(13.6％,1 453/10 688),A.baumannii(10.0％,1 067/10 688),and coagulase-negative Staphylococcus(9.9％,1 058/10 688).Multi-drug resistant organisms(MDROs)were very common in children.The prevalence of methicillin-resistant Staphylococcus aureus(MRSA),carbapenem-resistant Enterobacterales(CRE),carbapenem-resistant E.coli,carbapenem-resistant K.pneumoniae(CRKP),carbapenem-resistant A.baumannii(CRAB),and carbapenem-resistant P.aeruginosa(CRPA)was 41.1％,19.4％,8.8％,30.9％,67.4％,and 28.8％,respectively.Overall,more than 50％of Enterobacteriales isolates were resistant to cephalosporins,while nearly 25％of Enterobacteriales isolates were resistant to carbapenems.MDROs were highly resistant to commonly used antibiotics.More than 80％of CRE and CRAB strains were resistant to all beta-lactam antibiotics.CRE and CRAB showed low resistance rates to tigecycline and polymyxin.CRPA showed lower resistance rates to piperacillin,beta-lactamase inhibitor combinations than the resistance rates to third and fourth generation cephalosporins.All of the Staphylococcus and Enterococcus isolates were susceptible to vancomycin and tigecycline.None of PRSP strains isolated from meningitis and nonmeningitis samples were resistant to rifampicin,vancomycin,or linezolid.The prevalence of β-lactamase-negative ampicillin-resistant(BLNAR)strains was 43.3％in Haemophilus influenzae.Conclusions MDROs were prevalent in PICU.It is necessary to establish an effective multidisciplinary team(MDT)to control the antimicrobial resistance.

Objective:To explore the implementation effects of an proactive health management model for elderly patients with multimorbidity in comprehensive hospitals based on the concept of people-centered integrated care (PCIC).Methods:This study was a randomized controlled trial. Elderly patients who were hospitalized in the Department of General Practice at Shanghai East Hospital Tongji University and also suffered from hypertension, type 2 diabetes and dyslipidemia from November 2022 to January 2024 were included, and were divided into the control group (traditional health management, n=25) and the intervention group (proactive health management, n=25) using the random number table method. A research team comprising experts in general medicine, pharmacy, nutrition, rehabilitation medicine, psychology, and other relevant specialties was formed. Based on literature analysis, clinical experience, and hospital resources, the team collaborated to develop a comprehensive, hospital-based proactive health management model for elderly patients with comorbidities based on the PCIC concept. Patients in the control group were managed using the traditional health management model. Patients in the intervention group were managed using the proactive health management model. Baseline clinical data was collected and the patients were followed up for 6 months. At the 6-month follow-up, data on blood pressure, fasting blood glucose, and blood lipids were collected, as well as information on polypharmacy, activities of daily living (ADL) ability, 10-year atherosclerotic cardiovascular disease (ASCVD) risk, and unplanned rehospitalization were recorded. Results:A total of 50 patients were enrolled, with 25 patients in each group. The control group had an average age of (70.40±6.54) years, with 15 males(60.0%). The intervention group had an average age of (71.20±5.14) years, with 16 males(64.0%). At the 6-month follow-up, the standardization rates of blood pressure, fasting blood glucose, glycated hemoglobin, low-density lipoprotein cholesterol (LDL-C), non-high-density lipoprotein cholesterol (non-HDL-C), and triglycerides (TG) in both groups were higher than those in the baseline (all P<0.05).In addition, patients in the intervention group had the compliance rates for higher blood pressure, fasting blood glucose, 2-hour postprandial blood glucose, HbA1c, LDL-C, non-HDL-C, and TG than the control group (all P<0.05).At the 6-month follow-up, the 10-year ASCVD high-risk patient percentage decreased in the intervention group compared with baseline ( P=0.023) and was lower than that of the control group ( P=0.045), and the unanticipated readmission rate of patients in the intervention group was also lower than that of the control group ( P=0.042). Conclusions:A proactive health management model for elderly patients with geriatric multimorbidity in a comprehensive hospital, based on the concept of PCIC, was applied to an elderly population with concurrent hypertension, type 2 diabetes and dyslipidemia. The results of the management were favorable.

Objective To investigate the effects of STIP1 homology and U-box containing protein 1(STUB1)on the ubiquitination of glutathione peroxidase 4(GPX4)and ferroptosis in colon cancer cells HCT116,as well as the impact on CD8+T cell-mediated killing of HCT116 cells.Methods HCT116 cells were divided into control group,empty plasmid transfection(pcDNA3.1-vector)group,STUB1 overexpression plasmid transfection(pcDNA3.1-STUB1)group,and co-transfection(pcDNA3.1-STUB1+pcDNA3.1-GPX4)group.Cell proliferation ability was assessed by CCK-8 assay.Clonogenic ability was determined by clone formation assay.Malondialdehyde(MDA)levels in cells were measured using an MDA kit.Intracellular ferrous ion(Fe2+)levels were detected with an Fe2+probe.Changes in mitochondrial membrane potential were detected using JC-1 dye.Protein expression levels of STUB1,solute carrier family 7 member 11(SLC7A11),and GPX4 were determined by western blot.The binding between STUB1 and GPX4 was assessed by co-immunoprecipitation.The effect of STUB1 on GPX4 protein ubiquitination was detected using a ubiquitin antibody.HCT116 cells transfected with different plasmids were co-cultured with human peripheral blood CD8+T cells,and the killing ability of CD8+T cells against HCT116 cells was measured using a lactate dehydrogenase(LDH)kit.Perforin,granzyme,and interferon-γ levels in the co-culture supernatant were determined by ELISA.Results Compared with the control group,the pcDNA3.1-STUB1 group showed decreased cell proliferation ability,mitochondrial membrane potential,and protein expression levels of SLC7A11 and GPX4,along with increased STUB1 protein expression,MDA,Fe2+levels,and GPX4 ubiquitination in HCT116 cells.Compared with the pcDNA3.1-STUB1 group,the pcDNA3.1-STUB1+pcDNA3.1-GPX4 group exhibited increased cell proliferation ability,mitochondrial membrane potential,and expression levels of SLC7A11 and GPX4,along with decreased MDA and Fe2+levels in HCT116 cells.After co-culture of HCT116 cells with CD8+T cells,the pcDNA3.1-STUB1 group showed significantly increased killing rate of CD8+T cells against HCT116 cells,as well as elevated levels of perforin,granzyme,and interferon-γ in the co-culture supernatant compared with the control group.Compared with the pcDNA3.1-STUB1 group,the pcDNA3.1-STUB1+pcDNA3.1-GPX4 group exhibited decreased killing rate of CD8+T cells against HCT116 cells and reduced levels of perforin,granzyme,and interferon-γ in the co-culture supernatant.Conclusion Overexpression of STUB1 promotes GPX4 ubiquitination in colon cancer cells HCT116,induces ferroptosis,and enhances the killing effect of CD8+T cells on HCT116 cells.

Objective:To investigate the impact of olanzapine and risperidone on the cognitive function, sensory gating function and clinical symptoms of patients with first-episode schizophrenia(FES).Additionally, to analyze the correlation between sensory gating inhibitory deficits and cognitive impairment in FES patients.Methods:A total of 71 FES patients were selected in the Third People's Hospital of Zhongshan City from March 2023 to March 2024, and 60 healthy controls were recruited during the same period.The FES patients were divided into olanzapine group and risperidone group by random number table.Olanzapine group was treated with variable doses of olanzapine(10-20 mg/d), and risperidone group was treated with variable doses of risperidone(3-6 mg/d).The MATRICS consensus cognitive battery(MCCB) was used to evaluate the cognitive function of the patients, P50 index was measured by auditory paired condition-stimulus paradigm, and the efficacy was evaluated by positive and negative syndrome scale(PANSS) score reduction rate before and after 6 weeks of treatment.Healthy controls were assessed cognitive function only once with P50.SPSS 25.0 software was used for data processing. Perform statistical analysis using paired sample t-test, Wilcoxon signed rank test, independent sample t-test, Mann Whitney U test, χ2 test and generalized linear model. Results:Before treatment, the S2 amplitude of FES (1.74 (0.91, 2.79) μV) was higher than that of healthy controls (1.70 (1.04, 2.71) μV) (Wald χ2=4.483, P=0.034), the S2/S1 ratio of FES (0.58 (0.43, 0.78)) was higher than that of healthy controls (0.41 (0.31, 0.57)) (Wald χ2=10.909, P=0.001), and the difference of FES amplitude of S1-S2 was (1.22 (0.43, 1.92) μV) was lower than that of healthy controls (2.23 (1.54, 3.07) μV) (Wald χ2=17.679, P<0.001). There was no significant difference in PANSS, MCCB and P50 between olanzapine group and risperidone group before treatment (all P>0.05). After treatment, there was no significant difference in response rate between the two groups ( χ2=0.059, P=0.808), the PANSS scores were lower than those before treatment, the MCCB test results were higher than those before treatment (both P<0.05), and the P50 results were not statistically significant different compared with those before treatment (both P>0.05). The generalized linear model showed that the S1, S2 amplitude of the P50 had positive impact on the connection test score in the MCCB test ( β=0.466, P=0.020; β=0.879, P=0.009), other indicators were not found to have an impact on the test scores of the MCCB test (all P>0.05). Conclusion:Olanzapine and risperidone can significantly improve the cognitive function of FES, but the improvement of sensory gating deficits is limited. The pathogenic mechanism of sensory gating inhibitory deficits in FES may be different from that of cognitive dysfunction.

Objective:To investigate the impact of olanzapine and risperidone on the cognitive function, sensory gating function and clinical symptoms of patients with first-episode schizophrenia(FES).Additionally, to analyze the correlation between sensory gating inhibitory deficits and cognitive impairment in FES patients.Methods:A total of 71 FES patients were selected in the Third People's Hospital of Zhongshan City from March 2023 to March 2024, and 60 healthy controls were recruited during the same period.The FES patients were divided into olanzapine group and risperidone group by random number table.Olanzapine group was treated with variable doses of olanzapine(10-20 mg/d), and risperidone group was treated with variable doses of risperidone(3-6 mg/d).The MATRICS consensus cognitive battery(MCCB) was used to evaluate the cognitive function of the patients, P50 index was measured by auditory paired condition-stimulus paradigm, and the efficacy was evaluated by positive and negative syndrome scale(PANSS) score reduction rate before and after 6 weeks of treatment.Healthy controls were assessed cognitive function only once with P50.SPSS 25.0 software was used for data processing. Perform statistical analysis using paired sample t-test, Wilcoxon signed rank test, independent sample t-test, Mann Whitney U test, χ2 test and generalized linear model. Results:Before treatment, the S2 amplitude of FES (1.74 (0.91, 2.79) μV) was higher than that of healthy controls (1.70 (1.04, 2.71) μV) (Wald χ2=4.483, P=0.034), the S2/S1 ratio of FES (0.58 (0.43, 0.78)) was higher than that of healthy controls (0.41 (0.31, 0.57)) (Wald χ2=10.909, P=0.001), and the difference of FES amplitude of S1-S2 was (1.22 (0.43, 1.92) μV) was lower than that of healthy controls (2.23 (1.54, 3.07) μV) (Wald χ2=17.679, P<0.001). There was no significant difference in PANSS, MCCB and P50 between olanzapine group and risperidone group before treatment (all P>0.05). After treatment, there was no significant difference in response rate between the two groups ( χ2=0.059, P=0.808), the PANSS scores were lower than those before treatment, the MCCB test results were higher than those before treatment (both P<0.05), and the P50 results were not statistically significant different compared with those before treatment (both P>0.05). The generalized linear model showed that the S1, S2 amplitude of the P50 had positive impact on the connection test score in the MCCB test ( β=0.466, P=0.020; β=0.879, P=0.009), other indicators were not found to have an impact on the test scores of the MCCB test (all P>0.05). Conclusion:Olanzapine and risperidone can significantly improve the cognitive function of FES, but the improvement of sensory gating deficits is limited. The pathogenic mechanism of sensory gating inhibitory deficits in FES may be different from that of cognitive dysfunction.

Objective To investigate the effects of STIP1 homology and U-box containing protein 1(STUB1)on the ubiquitination of glutathione peroxidase 4(GPX4)and ferroptosis in colon cancer cells HCT116,as well as the impact on CD8+T cell-mediated killing of HCT116 cells.Methods HCT116 cells were divided into control group,empty plasmid transfection(pcDNA3.1-vector)group,STUB1 overexpression plasmid transfection(pcDNA3.1-STUB1)group,and co-transfection(pcDNA3.1-STUB1+pcDNA3.1-GPX4)group.Cell proliferation ability was assessed by CCK-8 assay.Clonogenic ability was determined by clone formation assay.Malondialdehyde(MDA)levels in cells were measured using an MDA kit.Intracellular ferrous ion(Fe2+)levels were detected with an Fe2+probe.Changes in mitochondrial membrane potential were detected using JC-1 dye.Protein expression levels of STUB1,solute carrier family 7 member 11(SLC7A11),and GPX4 were determined by western blot.The binding between STUB1 and GPX4 was assessed by co-immunoprecipitation.The effect of STUB1 on GPX4 protein ubiquitination was detected using a ubiquitin antibody.HCT116 cells transfected with different plasmids were co-cultured with human peripheral blood CD8+T cells,and the killing ability of CD8+T cells against HCT116 cells was measured using a lactate dehydrogenase(LDH)kit.Perforin,granzyme,and interferon-γ levels in the co-culture supernatant were determined by ELISA.Results Compared with the control group,the pcDNA3.1-STUB1 group showed decreased cell proliferation ability,mitochondrial membrane potential,and protein expression levels of SLC7A11 and GPX4,along with increased STUB1 protein expression,MDA,Fe2+levels,and GPX4 ubiquitination in HCT116 cells.Compared with the pcDNA3.1-STUB1 group,the pcDNA3.1-STUB1+pcDNA3.1-GPX4 group exhibited increased cell proliferation ability,mitochondrial membrane potential,and expression levels of SLC7A11 and GPX4,along with decreased MDA and Fe2+levels in HCT116 cells.After co-culture of HCT116 cells with CD8+T cells,the pcDNA3.1-STUB1 group showed significantly increased killing rate of CD8+T cells against HCT116 cells,as well as elevated levels of perforin,granzyme,and interferon-γ in the co-culture supernatant compared with the control group.Compared with the pcDNA3.1-STUB1 group,the pcDNA3.1-STUB1+pcDNA3.1-GPX4 group exhibited decreased killing rate of CD8+T cells against HCT116 cells and reduced levels of perforin,granzyme,and interferon-γ in the co-culture supernatant.Conclusion Overexpression of STUB1 promotes GPX4 ubiquitination in colon cancer cells HCT116,induces ferroptosis,and enhances the killing effect of CD8+T cells on HCT116 cells.

Objective:To explore the mediating role of coping strategies in the relationship between disease uncertainty and post-discharge coping difficulties in mothers of preterm infants.Methods:A convenience sampling method was used to select 255 mothers of preterm infants from Shandong Provincial Hospital Affiliated to Shandong First Medical University, Linyi People 's Hospital, Weifang Maternal and Child Health Hospital, Maternal and Child Care Center of Dezhou, Dongying People 's Hospital between March and May 2023. General information surveys, the Parents ' Perception of Uncertainty Scale, the Simple Coping Style Questionnaire, and Post-Discharge Coping Difficulty Scale-Parent Form were administered on the day of discharge and 3 weeks post-discharge. Structural equation modeling was constructed using AMOS 23.0 software, and the mediating effect was tested using the Bootstrap method. Results:The disease uncertainty score for the 255 mothers of preterm infants was 66.00 (53.00, 77.00), the active coping score was 23.00 (17.00, 27.00), the passive coping score was 10.00 (7.00, 13.00), and the post-discharge coping difficulties score was 42.00 (26.00, 55.00). Mediating effect analysis showed that active and passive coping partially mediated the relationship between disease uncertainty and post-discharge coping difficulties, with mediating effects of 18.73% and 17.06%, respectively.Conclusions:Disease uncertainty can indirectly affect post-discharge coping difficulties through active and passive coping strategies. Healthcare providers should assist mothers of preterm infants in appropriately managing disease uncertainty and actively coping with challenges.

To construct a recombinant fowl adenovirus 4(FAdV-4)expressing the Cap protein of goose astrovirus genotype 2(GoAstV-2),the expression cassette of Cap gene was inserted into the natural 1 966 bp deletion region of the FAdV-4 genome in the infectious clone p15A-cm-FAdV4-HNJZ.The resulted recombinant plasmid p15A-cm-FAdV4-HNJZ-Cap/GoAstV-2 was linearized with restriction enzyme and transfected into chicken hepatoma cell line(LMH)to rescue the recombinant FAdV-4 expressing the Cap protein of GoAstV-2,rF Ad V4-Cap/GoAstV-2.After 15 passages in LMH cells,the recombinant rFAdV4-Cap/GoAstV-2 was identified by PCR using primers flanking the insertion site of the Cap gene expression cassette and using viral genome DNA extracted from rFAdV4-Cap/GoAstV-2 infected LMH cells as template.LMH cells were in-fected with 15th passage rFAdV4-Cap/GoAstV-2 and indirect immunofluorescence was performed with a polyclonal antibody against Cap protein as the primary antibody.Western blot was carried out with lysates of rFAdV4-Cap/GoAstV-2 infected LMH cells.The in vitro replication dynamic of the 15th passage of the rFAdV4-Cap/GoAstV-2 was also investigated in LMH cells.The results demonstrated that the Cap gene of GoAstV-2 was presented in the genome of the recombinant vi-rus rF AdV4-Cap/Go Ast V-2,and could be expressed stably.The prepared recombinant virus in this study will lay a foundation for developing inactivated bivalent vaccine candidate against co-in-fection of FAdV-4 and GoAstV-2 in goose.

Objective This study aimed to investigate the antimicrobial resistance profiles of bacterial strains isolated from pediatric intensive care units(PICU)in China for better antimicrobial therapy.Methods Clinical isolates were collected from 17 institutions,including tertiary care children's hospitals and pediatric department of tertiary general hospitals in China from January 1,2020 to December 31,2022.Antimicrobial susceptibility testing was carried out according to a unified protocol using Kirby-Bauer method or automated systems.Results were interpreted according to the breakpoints released by the Clinical and Laboratory Standards Institute(CLSI)in 2020.Results A total of 10 688 isolates were collected,including gram-positive organisms(39.2％)and gram-negative organisms(60.8％).The top three organisms were S.aureus(13.6％,1 453/10 688),A.baumannii(10.0％,1 067/10 688),and coagulase-negative Staphylococcus(9.9％,1 058/10 688).Multi-drug resistant organisms(MDROs)were very common in children.The prevalence of methicillin-resistant Staphylococcus aureus(MRSA),carbapenem-resistant Enterobacterales(CRE),carbapenem-resistant E.coli,carbapenem-resistant K.pneumoniae(CRKP),carbapenem-resistant A.baumannii(CRAB),and carbapenem-resistant P.aeruginosa(CRPA)was 41.1％,19.4％,8.8％,30.9％,67.4％,and 28.8％,respectively.Overall,more than 50％of Enterobacteriales isolates were resistant to cephalosporins,while nearly 25％of Enterobacteriales isolates were resistant to carbapenems.MDROs were highly resistant to commonly used antibiotics.More than 80％of CRE and CRAB strains were resistant to all beta-lactam antibiotics.CRE and CRAB showed low resistance rates to tigecycline and polymyxin.CRPA showed lower resistance rates to piperacillin,beta-lactamase inhibitor combinations than the resistance rates to third and fourth generation cephalosporins.All of the Staphylococcus and Enterococcus isolates were susceptible to vancomycin and tigecycline.None of PRSP strains isolated from meningitis and nonmeningitis samples were resistant to rifampicin,vancomycin,or linezolid.The prevalence of β-lactamase-negative ampicillin-resistant(BLNAR)strains was 43.3％in Haemophilus influenzae.Conclusions MDROs were prevalent in PICU.It is necessary to establish an effective multidisciplinary team(MDT)to control the antimicrobial resistance.