ObjectiveTo investigate the epidemiological and clinical characteristics of human bocavirus (HBoV) in hospitalized children with acute lower respiratory tract infection (ALRTI) at a single-center children’s hospital in Shanghai, thereby providing evidence for the diagnosis, treatment, and prevention of HBoV infection. MethodsA retrospective study was conducted on 19 537 hospitalized children with ALRTI at Shanghai Children’s Hospital from January 2021 to December 2023. Multiplex polymerase chain reaction (PCR) combined with capillary electrophoresis was used to detect HBoV and 12 other common respiratory viruses /atypical pathogens. The positive detection rate, demographic characteristics (sex, age), temporal distribution (year, season) of HBoV, as well as the clinical characteristics of severe and non-severe pneumonia were analyzed. ResultsThe overall HBoV-positive rate was 2.57% (503/19 537), with 59.44% (299/503) being single infections and 40.56% (204/503) being co-infections. The positive detection rate was significantly higher in boys than that in girls (2.78% vs 2.33%, χ²=3.88, P=0.049). The highest infection rate was observed in toddlers, followed by infants (χ²=379.57, P<0.001). The positive rate peaked in 2021 and reached its lowest point in 2023 (χ²=45.49, P<0.001), with epidemics mainly prevalent in summer and autumn. The main clinical symptoms were cough (90.06%, 453/503), fever (75.94%, 382/503), and wheezing (39.96%, 201/503). Children with severe pneumonia showed a higher incidence of wheezing compared with the non-severe group (P<0.001), while underlying diseases and co-infections had no significant association with disease severity (P>0.05). ConclusionHBoV was an important pathogen of ALRTI in children, predominantly affecting infants and toddlers, with higher susceptibility in boys and seasonal peaks in autumn and summer. The main clinical manifestations included cough, fever, and wheezing, with wheezing being more prevalent in children with severe pneumonia.

Objective To investigate the molecular mechanism of Xiaoding Ointment in inhibiting pyroptosis through nucle-otide binding oligomerization of domain like receptor protein 3(NLRP3)/nuclear factor kappa B(NF-κB)/caspase-1 pathway,and to analyze its therapeutic effect on acute closed soft tissue injury.Methods Sixty SPF SD rats were selected and randomly divided into four groups:blank group,model group,positive control group(Qizheng Xiaotong plaster),and Xiaoding Ointment group(topical application).The blank group did not receive any treatment,while the other groups received corresponding inter-vention treatment after preparing the acute closed soft tissue injury model,with a treatment period of seven days.The conditions such as swelling,pain and limited movement were evaluated through the injury symptom score.Hematoxylin-eosin(HE)staining was used to observe the histopathological changes of rat leg muscle tissue.ELISA was used to detect the levels of serum inflam-matory factors IL-1β and TNF-α.TUNEL staining was used to observe the pyroptosis of cells.The expression levels of NLRP3,NF-κB,p65 and Caspase-1 proteins were analyzed by Western blotting and immunohistochemistry to evaluate the therapeutic effect and mechanism of action of Xiaoding Ointment.Results Compared with the model group,the swelling degree,appearance changes,activity and pain behavior of rats in the Xiaoding Ointment group were significantly improved(all P＜0.05).Histopa-thology showed a decrease in inflammatory cell infiltration and a normalization of muscle fiber arrangement.The levels of serum inflammatory factors IL-1β and TNF-α were significantly reduced(both P＜0.05);The number of pyroptosis positive cells and pyroptosis positive rate significantly decreased(both P＜0.05);The protein expressions of NLRP3,NF-κB,p65,and Caspase-1 was significantly reduced(all P＜0.05).Conclusion Xiaoding Ointment inhibits NLRP3 inflammasome activation and NF-κB signaling pathway through targeted inhibition,blocks Caspase-1-dependent apoptosis process,and reduces the release of inflam-matory factors such as IL-1β and TNF-α,thereby alleviating the inflammatory response after acute soft tissue injury and promo-ting tissue repair.

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 molecular mechanism of Xiaoding Ointment in inhibiting pyroptosis through nucle-otide binding oligomerization of domain like receptor protein 3(NLRP3)/nuclear factor kappa B(NF-κB)/caspase-1 pathway,and to analyze its therapeutic effect on acute closed soft tissue injury.Methods Sixty SPF SD rats were selected and randomly divided into four groups:blank group,model group,positive control group(Qizheng Xiaotong plaster),and Xiaoding Ointment group(topical application).The blank group did not receive any treatment,while the other groups received corresponding inter-vention treatment after preparing the acute closed soft tissue injury model,with a treatment period of seven days.The conditions such as swelling,pain and limited movement were evaluated through the injury symptom score.Hematoxylin-eosin(HE)staining was used to observe the histopathological changes of rat leg muscle tissue.ELISA was used to detect the levels of serum inflam-matory factors IL-1β and TNF-α.TUNEL staining was used to observe the pyroptosis of cells.The expression levels of NLRP3,NF-κB,p65 and Caspase-1 proteins were analyzed by Western blotting and immunohistochemistry to evaluate the therapeutic effect and mechanism of action of Xiaoding Ointment.Results Compared with the model group,the swelling degree,appearance changes,activity and pain behavior of rats in the Xiaoding Ointment group were significantly improved(all P＜0.05).Histopa-thology showed a decrease in inflammatory cell infiltration and a normalization of muscle fiber arrangement.The levels of serum inflammatory factors IL-1β and TNF-α were significantly reduced(both P＜0.05);The number of pyroptosis positive cells and pyroptosis positive rate significantly decreased(both P＜0.05);The protein expressions of NLRP3,NF-κB,p65,and Caspase-1 was significantly reduced(all P＜0.05).Conclusion Xiaoding Ointment inhibits NLRP3 inflammasome activation and NF-κB signaling pathway through targeted inhibition,blocks Caspase-1-dependent apoptosis process,and reduces the release of inflam-matory factors such as IL-1β and TNF-α,thereby alleviating the inflammatory response after acute soft tissue injury and promo-ting tissue repair.

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 value of tumor volume to uterine volume ratio (N/U) combined with the expression of alpha-fetoprotein (AFP), sugar antigen 199 (CA199) and human epididymal secretory protein 4 (HE4) in evaluating the pathologic grade and prognosis of endometrial carcinoma (EC) .Methods:A total of 160 EC patients admitted to Linyi Central Hospital from Jan. 2021 to Dec. 2023 were divided into low-grade group and high-grade group according to FIGO grading method, and were divided into poor prognosis group and good prognosis group according to cancer death, recrudescence. The levels of N/U, AFP, CA199 and HE4 in patients with different pathologic grades and prognosis were compared. COX regression was used to analyze the influencing factors of EC adverse prognosis, ROC curve was used to analyze the value of N/U combined with serum AFP, CA199 and HE4 in predicting EC adverse prognosis, and a nomogram model was constructed.Results:Pathological examination of 160 EC patients showed that 12 cases were non-endometrioid adenocarcinoma, 148 cases were endometrioid adenocarcinoma, 41 cases were high-grade and 119 cases were low-grade.According to the follow-up, 94 of the 160 EC patients had good prognosis and 66 had poor prognosis. The levels of N/U, AFP, CA199 and HE4 in the poor prognosis group were higher than those in the good prognosis group ( P<0.05). COX regression analysis showed that high levels of N/U, AFP, CA199 and HE4 were all factors affecting the poor prognosis of EC patients ( P<0.05). The AUC value of combined detection of N/U, AFP, CA199 and HE4 in predicting adverse prognosis of EC patients was higher than that of single detection ( Z=3.140, 3.658, 4.277, 4.378, P<0.05) .The ROC curve AUC (95% CI) of the training set and the validation set were 0.84 (0.77-0.92) and 0.90 (0.81-0.98) respectively for the training set and the validation set to predict the adverse prognosis of EC patients. Calibration curve results showed that the calibration curve for EC patients predicted by the nomogram model was close to the ideal curve ( P=0.521, 0.743). The DCA curve shows that the probability threshold of the nomogram model has a higher positive net return at 20%~100%. Conclusion:The levels of N/U, AFP, CA199 and HE4 in EC patients are related to the pathologic grade, and the combined detection of these indicators can predict the poor prognosis of EC patients, and the nomogram model constructed based on these indicators has high predictive value.

Objective:To investigate the value of tumor volume to uterine volume ratio (N/U) combined with the expression of alpha-fetoprotein (AFP), sugar antigen 199 (CA199) and human epididymal secretory protein 4 (HE4) in evaluating the pathologic grade and prognosis of endometrial carcinoma (EC) .Methods:A total of 160 EC patients admitted to Linyi Central Hospital from Jan. 2021 to Dec. 2023 were divided into low-grade group and high-grade group according to FIGO grading method, and were divided into poor prognosis group and good prognosis group according to cancer death, recrudescence. The levels of N/U, AFP, CA199 and HE4 in patients with different pathologic grades and prognosis were compared. COX regression was used to analyze the influencing factors of EC adverse prognosis, ROC curve was used to analyze the value of N/U combined with serum AFP, CA199 and HE4 in predicting EC adverse prognosis, and a nomogram model was constructed.Results:Pathological examination of 160 EC patients showed that 12 cases were non-endometrioid adenocarcinoma, 148 cases were endometrioid adenocarcinoma, 41 cases were high-grade and 119 cases were low-grade.According to the follow-up, 94 of the 160 EC patients had good prognosis and 66 had poor prognosis. The levels of N/U, AFP, CA199 and HE4 in the poor prognosis group were higher than those in the good prognosis group ( P<0.05). COX regression analysis showed that high levels of N/U, AFP, CA199 and HE4 were all factors affecting the poor prognosis of EC patients ( P<0.05). The AUC value of combined detection of N/U, AFP, CA199 and HE4 in predicting adverse prognosis of EC patients was higher than that of single detection ( Z=3.140, 3.658, 4.277, 4.378, P<0.05) .The ROC curve AUC (95% CI) of the training set and the validation set were 0.84 (0.77-0.92) and 0.90 (0.81-0.98) respectively for the training set and the validation set to predict the adverse prognosis of EC patients. Calibration curve results showed that the calibration curve for EC patients predicted by the nomogram model was close to the ideal curve ( P=0.521, 0.743). The DCA curve shows that the probability threshold of the nomogram model has a higher positive net return at 20%~100%. Conclusion:The levels of N/U, AFP, CA199 and HE4 in EC patients are related to the pathologic grade, and the combined detection of these indicators can predict the poor prognosis of EC patients, and the nomogram model constructed based on these indicators has high predictive value.

Objectives: . Head and neck squamous cell carcinoma (HNSCC) exhibits high recurrence rates, particularly in cases of radioresistant HNSCC (RR-HNSCC). Non-thermal plasma (NTP) therapy effectively suppresses the progression of HNSCC. However, the therapeutic mechanisms of NTP therapy in treating RR-HNSCC are not well understood. In this study, we explored the regulatory role of NTP in the RR-HNSCC signaling pathway and identified its signature genes. Methods: . After constructing two RR-HNSCC cell lines, we prepared cell lysates from cells treated or not treated with NTP-activated media (NTPAM) and performed RNA sequencing to determine their mRNA expression profiles. Based on the RNA sequencing results, we identified differentially expressed genes (DEGs), followed by a bioinformatics analysis to identify candidate molecules potentially associated with NTPAM therapy for RR-HNSCC. Results: . NTPAM reduced RR-HNSCC cell viability in vitro. RNA sequencing results indicated that NTPAM treatment activated the reactive oxygen species (ROS) pathway and induced ferroptosis in RR-HNSCC cell lines. Among the 1,924 genes correlated with radiation treatment, eight showed statistical significance in both the cell lines and The Cancer Genome Atlas (TCGA) cohort. Only five genes—ABCC3, DUSP16, PDGFB, RAF1, and THBS1—showed consistent results between the NTPAM data sequencing and TCGA data. LASSO regression analysis revealed that five genes were associated with cancer prognosis, with a hazard ratio of 2.26. In RR-HNSCC cells, NTPAM affected DUSP16, PDGFB, and THBS1 as activated markers within 6 hours, and this effect persisted for 12 hours. Furthermore, enrichment analysis indicated that these three DEGs were associated with the extracellular matrix, transforming growth factor-beta, phosphoinositide 3-kinase/protein kinase B, and mesenchymal-epithelial transition factor pathways. Conclusion . NTPAM therapy exerts cytotoxic effects in RR-HNSCC cell lines by inducing specific ROS-mediated ferroptosis. DUSP16, PDGFB, and THBS1 were identified as crucial targets for reversing the radiation resistance induced by NTPAM therapy, providing insights into the mechanisms and clinical applications of NTPAM treatment in RR-HNSCC.

Objectives: . Head and neck squamous cell carcinoma (HNSCC) exhibits high recurrence rates, particularly in cases of radioresistant HNSCC (RR-HNSCC). Non-thermal plasma (NTP) therapy effectively suppresses the progression of HNSCC. However, the therapeutic mechanisms of NTP therapy in treating RR-HNSCC are not well understood. In this study, we explored the regulatory role of NTP in the RR-HNSCC signaling pathway and identified its signature genes. Methods: . After constructing two RR-HNSCC cell lines, we prepared cell lysates from cells treated or not treated with NTP-activated media (NTPAM) and performed RNA sequencing to determine their mRNA expression profiles. Based on the RNA sequencing results, we identified differentially expressed genes (DEGs), followed by a bioinformatics analysis to identify candidate molecules potentially associated with NTPAM therapy for RR-HNSCC. Results: . NTPAM reduced RR-HNSCC cell viability in vitro. RNA sequencing results indicated that NTPAM treatment activated the reactive oxygen species (ROS) pathway and induced ferroptosis in RR-HNSCC cell lines. Among the 1,924 genes correlated with radiation treatment, eight showed statistical significance in both the cell lines and The Cancer Genome Atlas (TCGA) cohort. Only five genes—ABCC3, DUSP16, PDGFB, RAF1, and THBS1—showed consistent results between the NTPAM data sequencing and TCGA data. LASSO regression analysis revealed that five genes were associated with cancer prognosis, with a hazard ratio of 2.26. In RR-HNSCC cells, NTPAM affected DUSP16, PDGFB, and THBS1 as activated markers within 6 hours, and this effect persisted for 12 hours. Furthermore, enrichment analysis indicated that these three DEGs were associated with the extracellular matrix, transforming growth factor-beta, phosphoinositide 3-kinase/protein kinase B, and mesenchymal-epithelial transition factor pathways. Conclusion . NTPAM therapy exerts cytotoxic effects in RR-HNSCC cell lines by inducing specific ROS-mediated ferroptosis. DUSP16, PDGFB, and THBS1 were identified as crucial targets for reversing the radiation resistance induced by NTPAM therapy, providing insights into the mechanisms and clinical applications of NTPAM treatment in RR-HNSCC.

Objectives: . Head and neck squamous cell carcinoma (HNSCC) exhibits high recurrence rates, particularly in cases of radioresistant HNSCC (RR-HNSCC). Non-thermal plasma (NTP) therapy effectively suppresses the progression of HNSCC. However, the therapeutic mechanisms of NTP therapy in treating RR-HNSCC are not well understood. In this study, we explored the regulatory role of NTP in the RR-HNSCC signaling pathway and identified its signature genes. Methods: . After constructing two RR-HNSCC cell lines, we prepared cell lysates from cells treated or not treated with NTP-activated media (NTPAM) and performed RNA sequencing to determine their mRNA expression profiles. Based on the RNA sequencing results, we identified differentially expressed genes (DEGs), followed by a bioinformatics analysis to identify candidate molecules potentially associated with NTPAM therapy for RR-HNSCC. Results: . NTPAM reduced RR-HNSCC cell viability in vitro. RNA sequencing results indicated that NTPAM treatment activated the reactive oxygen species (ROS) pathway and induced ferroptosis in RR-HNSCC cell lines. Among the 1,924 genes correlated with radiation treatment, eight showed statistical significance in both the cell lines and The Cancer Genome Atlas (TCGA) cohort. Only five genes—ABCC3, DUSP16, PDGFB, RAF1, and THBS1—showed consistent results between the NTPAM data sequencing and TCGA data. LASSO regression analysis revealed that five genes were associated with cancer prognosis, with a hazard ratio of 2.26. In RR-HNSCC cells, NTPAM affected DUSP16, PDGFB, and THBS1 as activated markers within 6 hours, and this effect persisted for 12 hours. Furthermore, enrichment analysis indicated that these three DEGs were associated with the extracellular matrix, transforming growth factor-beta, phosphoinositide 3-kinase/protein kinase B, and mesenchymal-epithelial transition factor pathways. Conclusion . NTPAM therapy exerts cytotoxic effects in RR-HNSCC cell lines by inducing specific ROS-mediated ferroptosis. DUSP16, PDGFB, and THBS1 were identified as crucial targets for reversing the radiation resistance induced by NTPAM therapy, providing insights into the mechanisms and clinical applications of NTPAM treatment in RR-HNSCC.