Objective:To investigate the clinical characteristics of children with severe human metapneumovirus (HMPV) infection and identify the risk factors associated with critical illness.Methods:A retrospective cohort study was conducted, enrolling 157 hospitalized children with severe HMPV infection, who tested positive for HMPV nucleic acid via PCR-capillary electrophoresis fragment analysis of nasopharyngeal secretions at Shanghai Children′s Hospital from January 2021 to December 2023.Clinical features, co-infections, treatment, and outcomes were collected. Based on the diagnostic criteria for severe HMPV infection, the patients were categorized into a critical illness group and a non-critical illness group. Intergroup comparisons were performed using the χ2 test or the Mann-Whitney U test. Multivariate Logistic regression analysis was employed to identify risk factors for critical HMPV infection and to establish a predictive model.The performance of the model was evaluated using receiver operating characteristic (ROC) curve analysis and calibration curves. Results:Among the 157 cases of severe HMPV infection, there were 67 males and 90 females, with an onset age of 39.0 (20.0, 55.5) months. Single-pathogen infection was observed in 125 cases (79.6%), while mixed infections accounted for 32 cases (20.4%).Severe pneumonia was diagnosed in 136 cases (86.6%).The predominant manifestations of severe HMPV infection included fever 152 cases (96.8%), cough 151 cases (96.2%), and wheezing 94 cases (59.9%).Sixty-eight patients (43.3%) required non-invasive respiratory support, 58 cases (36.9%) were admitted to the intensive care unit, and 22 cases (14.0%) underwent mechanical ventilation. Of the total, 149 cases (94.9%) were discharged with improvement, 8 cases (5.1%) were discharged against medical advice, and there were no fatal cases. The cohort was further stratified into a critical illness group 31 cases and a non-critical illness group 126 cases. Compared to the non-critical illness group, the critical illness group exhibited significantly higher rates of respiratory distress, lethargy, and intercostal retractions, along with a higher proportion of underlying comorbidities, and elevated levels of C-reactive protein and procalcitonin (all P<0.05).Conversely, albumin and hemoglobin levels were significantly lower in the critical illness group (both P<0.05). ROC curve analysis revealed that the optimal cutoff value for the duration of fever in predicting severe HMPV infection was 4.5 days.The multivariate binary Logistic regression analysis revealed that prolonged fever duration (>4.5 days) ( OR=28.00, 95% CI 5.09-153.93, P<0.001), anorexia ( OR=11.72, 95% CI 1.26-108.75, P=0.030), and immune dysfunction ( OR=36.71, 95% CI 1.55-867.31, P=0.026) were independent risk factors for severe HMPV infection. A predictive model for critical illness was constructed based on these independent risk factors. ROC curve analysis demonstrated excellent discriminative ability, with an area under the curve of 0.96 (95% CI 0.92-1.00, P<0.001). The optimal predictive probability threshold was 0.17, yielding a sensitivity of 0.93 and specificity of 0.92. The calibration curve closely approximated the ideal curve, indicating good model calibration ( P=0.157). Conclusions:Severe HMPV infection is predominantly observed as a single infection and is prone to progress to severe pneumonia, with fever, cough, and wheezing as the main clinical manifestations. A subset of cases progresses to critical illness, though the overall prognosis is favorable. Prolonged fever duration (>4.5 days), anorexia, and immune dysfunction were independent risk factors for critical illness.The risk prediction model constructed for pediatric critical HMPV infection demonstrated robust discriminative ability with excellent calibration.

Objective:To analyze the distribution and epidemiological characteristics of non-bacterial pathogens in hospitalized children with acute respiratory infections at a tertiary pediatric hospital in Shanghai during 2023.Methods:A retrospective study was conducted on 10 591 children with acute respiratory tract infections who were hospitalized in Shanghai Children's Hospital from January to December 2023. A multiplex PCR combined with capillary electrophoresis platform was used to detect 11 common non-bacterial respiratory pathogens（including viruses and atypical pathogens）. Statistical analysis was carried out using SPSS 29.0 software. Qualitative data were presented as numbers and percentages，and the Chi-square test was employed to make comparisons between groups，aiming to analyze the differences in the distribution of different pathogens according to gender，age group，and season. Additionally，based on the severity of the disease，patients were calssified into a severe pneumonia group and a non-severe pneumonia group to further explore the characteristics of the pathogen spectrum of severe pneumonia.Results:The total detection rate of pathogens was 54.39%（5 760/10 591），and the proportion of mixed infections was 12.76%（735/5 760）. The dominant pathogens and their proportions were as follows： Mycoplasma pneumoniae（19.20%，2 034/10 591），human rhinovirus（12.16%，1 288/10 591），influenza A virus（8.31%，880/10 591），and respiratory syncytial virus（8.14%，862/10 591）. Epidemiological characteristics showed that：（1）In terms of age： Mycoplasma pneumoniae was more common in older children（29.55%，901/3 049，in the school-age group，χ 2 = 653.67， P<0.001）. Influenza A virus had a high incidence in the adolescent group（11.34%，45/397，χ 2=48.69， P<0.001）. Respiratory syncytial virus was most susceptible in the infant group（20.94%，280/1 337，χ 2=739.92， P<0.001）. Human rhinovirus showed the characteristic of general susceptibility across all ages.（2）Monthly and seasonal distribution： Mycoplasma pneumoniae had a seasonal epidemic in summer and autumn（it began to rise in May and peaked in October at 34.22%，439/1 283）；influenza A virus had a bimodal distribution in spring and winter（the peak was 37.15% in March，315/848）；respiratory syncytial virus had a dominant epidemic in spring and summer（the detection rate was 21.24% in May，206/970），and human rhinovirus was prevalent throughout the year.（3）Clinical correlation：The detection rate of pathogens in the severe pneumonia group was significantly higher than that in the non-severe group：84.19%（426/506） vs 2.89%（5 334/10 085），χ 2=56.23， P<0.001. Conclusions:In 2023，the pathogen spectrum of hospitalized children with acute respiratory infections in the Shanghai area exhibits an epidemic pattern dominated by Mycoplasma pneumoniae，and its transmission dynamics are significantly age-dependent. This study delineates the pathogen-host-environment tripartite interactions，establishing an evidence-based foundation for formulating precision diagnostic-therapeutic algorithms and seasonal nosocomial infection prevention frameworks.

Objective:To analyze the distribution and epidemiological characteristics of non-bacterial pathogens in hospitalized children with acute respiratory infections at a tertiary pediatric hospital in Shanghai during 2023.Methods:A retrospective study was conducted on 10 591 children with acute respiratory tract infections who were hospitalized in Shanghai Children's Hospital from January to December 2023. A multiplex PCR combined with capillary electrophoresis platform was used to detect 11 common non-bacterial respiratory pathogens（including viruses and atypical pathogens）. Statistical analysis was carried out using SPSS 29.0 software. Qualitative data were presented as numbers and percentages，and the Chi-square test was employed to make comparisons between groups，aiming to analyze the differences in the distribution of different pathogens according to gender，age group，and season. Additionally，based on the severity of the disease，patients were calssified into a severe pneumonia group and a non-severe pneumonia group to further explore the characteristics of the pathogen spectrum of severe pneumonia.Results:The total detection rate of pathogens was 54.39%（5 760/10 591），and the proportion of mixed infections was 12.76%（735/5 760）. The dominant pathogens and their proportions were as follows： Mycoplasma pneumoniae（19.20%，2 034/10 591），human rhinovirus（12.16%，1 288/10 591），influenza A virus（8.31%，880/10 591），and respiratory syncytial virus（8.14%，862/10 591）. Epidemiological characteristics showed that：（1）In terms of age： Mycoplasma pneumoniae was more common in older children（29.55%，901/3 049，in the school-age group，χ 2 = 653.67， P<0.001）. Influenza A virus had a high incidence in the adolescent group（11.34%，45/397，χ 2=48.69， P<0.001）. Respiratory syncytial virus was most susceptible in the infant group（20.94%，280/1 337，χ 2=739.92， P<0.001）. Human rhinovirus showed the characteristic of general susceptibility across all ages.（2）Monthly and seasonal distribution： Mycoplasma pneumoniae had a seasonal epidemic in summer and autumn（it began to rise in May and peaked in October at 34.22%，439/1 283）；influenza A virus had a bimodal distribution in spring and winter（the peak was 37.15% in March，315/848）；respiratory syncytial virus had a dominant epidemic in spring and summer（the detection rate was 21.24% in May，206/970），and human rhinovirus was prevalent throughout the year.（3）Clinical correlation：The detection rate of pathogens in the severe pneumonia group was significantly higher than that in the non-severe group：84.19%（426/506） vs 2.89%（5 334/10 085），χ 2=56.23， P<0.001. Conclusions:In 2023，the pathogen spectrum of hospitalized children with acute respiratory infections in the Shanghai area exhibits an epidemic pattern dominated by Mycoplasma pneumoniae，and its transmission dynamics are significantly age-dependent. This study delineates the pathogen-host-environment tripartite interactions，establishing an evidence-based foundation for formulating precision diagnostic-therapeutic algorithms and seasonal nosocomial infection prevention frameworks.

Objective:To investigate the clinical characteristics of children with severe human metapneumovirus (HMPV) infection and identify the risk factors associated with critical illness.Methods:A retrospective cohort study was conducted, enrolling 157 hospitalized children with severe HMPV infection, who tested positive for HMPV nucleic acid via PCR-capillary electrophoresis fragment analysis of nasopharyngeal secretions at Shanghai Children′s Hospital from January 2021 to December 2023.Clinical features, co-infections, treatment, and outcomes were collected. Based on the diagnostic criteria for severe HMPV infection, the patients were categorized into a critical illness group and a non-critical illness group. Intergroup comparisons were performed using the χ2 test or the Mann-Whitney U test. Multivariate Logistic regression analysis was employed to identify risk factors for critical HMPV infection and to establish a predictive model.The performance of the model was evaluated using receiver operating characteristic (ROC) curve analysis and calibration curves. Results:Among the 157 cases of severe HMPV infection, there were 67 males and 90 females, with an onset age of 39.0 (20.0, 55.5) months. Single-pathogen infection was observed in 125 cases (79.6%), while mixed infections accounted for 32 cases (20.4%).Severe pneumonia was diagnosed in 136 cases (86.6%).The predominant manifestations of severe HMPV infection included fever 152 cases (96.8%), cough 151 cases (96.2%), and wheezing 94 cases (59.9%).Sixty-eight patients (43.3%) required non-invasive respiratory support, 58 cases (36.9%) were admitted to the intensive care unit, and 22 cases (14.0%) underwent mechanical ventilation. Of the total, 149 cases (94.9%) were discharged with improvement, 8 cases (5.1%) were discharged against medical advice, and there were no fatal cases. The cohort was further stratified into a critical illness group 31 cases and a non-critical illness group 126 cases. Compared to the non-critical illness group, the critical illness group exhibited significantly higher rates of respiratory distress, lethargy, and intercostal retractions, along with a higher proportion of underlying comorbidities, and elevated levels of C-reactive protein and procalcitonin (all P<0.05).Conversely, albumin and hemoglobin levels were significantly lower in the critical illness group (both P<0.05). ROC curve analysis revealed that the optimal cutoff value for the duration of fever in predicting severe HMPV infection was 4.5 days.The multivariate binary Logistic regression analysis revealed that prolonged fever duration (>4.5 days) ( OR=28.00, 95% CI 5.09-153.93, P<0.001), anorexia ( OR=11.72, 95% CI 1.26-108.75, P=0.030), and immune dysfunction ( OR=36.71, 95% CI 1.55-867.31, P=0.026) were independent risk factors for severe HMPV infection. A predictive model for critical illness was constructed based on these independent risk factors. ROC curve analysis demonstrated excellent discriminative ability, with an area under the curve of 0.96 (95% CI 0.92-1.00, P<0.001). The optimal predictive probability threshold was 0.17, yielding a sensitivity of 0.93 and specificity of 0.92. The calibration curve closely approximated the ideal curve, indicating good model calibration ( P=0.157). Conclusions:Severe HMPV infection is predominantly observed as a single infection and is prone to progress to severe pneumonia, with fever, cough, and wheezing as the main clinical manifestations. A subset of cases progresses to critical illness, though the overall prognosis is favorable. Prolonged fever duration (>4.5 days), anorexia, and immune dysfunction were independent risk factors for critical illness.The risk prediction model constructed for pediatric critical HMPV infection demonstrated robust discriminative ability with excellent calibration.

Objective:To investigate the efficacy and prognosis of CLAG±DAC (Clofarabine, Cytarabine, G-CSF±Decitabine) chemotherapy in patients with relapsed/refractory acute myeloid leukemia (R/R AML) .Methods:Continuous cases of R/R AML treated with the CLAG+DAC protocol or CLAG alone at the First Affiliated Hospital of Soochow University from January 2017 to December 2021 were retrospectively analyzed. The baseline characteristics, individual treatment regimen, treatment effect, disease progression, and survival status of patients were recorded. The factors influencing the efficacy of the CLAG±DAC chemotherapy regimens were analyzed, and the overall survival (OS) time after reinduction was calculated using the Kaplan-Meier method.Results:This study included a total of 53 patients, with 33 male patients and an average age of 40.6 years. Thirty-three patients achieved complete remission (CR+CRi) of the disease after the CLAG±DAC chemotherapy regimen and six patients achieved partial remission (PR), while 14 did not. Thirty-two patients eventually underwent hematopoietic stem cell transplantation, and the median OS of the patients was 55.9 months until follow-up. Patients with disease remission after the application of the CLAG±DAC chemotherapy had a significantly longer survival time than those without remission ( P<0.001). The results of the multifactorial analysis have revealed that combined DAC ( OR=4.60, 95% CI 1.14-23.5, P=0.04) and DNMT3A mutation ( OR=0.14, 95% CI 0.01-0.89, P=0.05) were the factors influencing the efficacy of the CLAG±DAC chemotherapy regimen. The remission rate was relatively higher in patients with R/R AML combined with FLT3-ITD mutation by applying the DAC+CLAG regimen ( OR=10.84, 95% CI 1.48-288.50, P=0.04) . Conclusion:The CLAG±DAC regimen is considered effective in patients with R/R AML, whereas decitabine combined with the CLAG regimen is more suitable for patients with R/R AML combined with FLT3-ITD mutation.

A simple, fast, and visual method for detecting antibodies against peste des petits ruminants virus (PPRV) using colloidal gold strips was developed. In this study, the pET-32a-N was transformed into Escherichia coli Rosetta (DE3) for expression. Hybridoma cell lines were generated by fusing SP2/0 myeloma cells with splenocytes from immunized mice with the expressed and purified N protein of PPRV. The PPRV N protein was labeled with colloidal gold particles as the gold-labeled antigen. The N protein served as the gold standard antigen and as the test (T) line-coated antigen, while the monoclonal antibody served as the quality control (C) line-coated antibody to assemble the colloidal gold immunochromatographic test strips for detecting antibodies against the N protein of PPRV. Hybridoma cell line designated as 1F1 was able to stably secrete the monoclonal antibody against the N protein of PPRV. The titer of 1F1 monoclonal antibody in ascites was 1:128 000 determined by indirect enzyme-linked immunosorbent assays (ELISA), and the immunoglobulin subtype of the monoclonal antibody was IgG1, with kappa chain. The obtained monoclonal antibody was able to specifically recognize the N protein of PPRV, as shown by Western blotting and indirect immunofluorescent assay (IFA). The developed colloidal gold test strip method was able to detect PPRV antibodies specifically, and there was no difference between different batches of the test strips. Testing of a total of 122 clinical sera showed that the compliance rate of the test strip with ELISA test was 97.6%.The test strip assay developed in this study has good specificity, reproducibility, and sensitivity, and it can be used for the rapid detection of PPRV antibodies.
Animals ; Mice ; Peste-des-Petits-Ruminants/prevention & control* ; Antibodies, Monoclonal ; Reproducibility of Results ; Peste-des-petits-ruminants virus ; Antibodies, Viral ; Enzyme-Linked Immunosorbent Assay ; Goats

Objective Acute pancreatitis exhibits different clinical and ultrasonic features in patients complicated with acute acalculous cholecystitis ( AAC) at different stages .The aim of this study was to analyze the ultrasonic characteristics of acute pancreati-tis complicated with AAC at different stages . Methods We retrospectively analyzed the clinical data about 41 cases of acute pancrea-titis with moderate to severe AAC .According to whether AAC developed within or after 2 weeks of the onset of acute pancreatitis , we divided the patients into an early-stage group (n=18) and a late-stage group (n=23).We recorded the gallbladder size, gallbladder wall thickness , fluid around the gallbladder , biliary sludge deposition and the Murphy′s sign by ultrasonography , obtained AAC-related clinical and laboratory data concerning body temperature , Murphy′s sign, WBC count and C-reactive protein level , and analyzed the ultrasonic features of AAC at different stages in the acute pancreatitis patients. Results All the patients experienced a fever of >38.5℃, 38.89%with chills in the early onset group and 47.83%in the late onset group .Increases were observed in patients of the early-and late-stage groups in the WBC count ( 94.44%vs 82.61%) , the C-reactive protein level ( 100%vs 91.30%) , and the fluid volume around the gallbladder (94.44%vs 60.86%, P<0.05), but incidence rate of gallbladder wall thickening was significantly lower in the former than in the latter group (11.11%vs 78.26%, P<0.01). Conclusion AAC developing at different stages of acute pancreatitis has different ultrasonic features , with higher incidence rates of fluid around the gallbladder in the early stage and gallbladder wall thickening in the late stage.