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.

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.

Objective:To analyze the distribution of tumor necrosis factor-alpha（TNF-α）308 gene loci polymorphism in children with febrile seizures（FS）and to explore the correlation between TNF-α 308 gene polymorphisms and FS in children.Methods:A total of 320 children diagnosed with FS in the Department of Emergency，Shanghai Children's Hospital from September 1st，2020 to June 30th，2021 were enrolled as the study subjects，which were divided into simple febrile seizures（SFS）group（232 cases）and complex febrile seizures（CFS）group（88 cases）based on their clinical characteristics，and the clinical characteristics and laboratory indexes of the two groups were compared. Children with no history of convulsions were selected as the control group（160 cases）. The high-resolution melting and gene sequencing technology were used to analyze the polymorphism of TNF-α 308 gene in each group and the distribution of different gene types and allele frequencies among the groups was compared. A multivariate Logistic regression model was constructed to analyze the relationship between TNF-α 308 gene polymorphism and FS.Results:The age，mean corpuscular volume，mean corpuscular hemoglobin and platelet distribution width of the CFS group were significantly higher than those in the SFS group，and the difference was statistically significant（all P<0.05）.There was no significant difference in gender distribution，family history of FS，history of FS，body temperature at time of convulsions，WBC，Hb，CRP and PLT between the two groups（all P>0.05）.The genotype frequency distribution of TNF-α 308 polymorphism in the three groups was in line with the Hardy-Weinberg equilibrium（ P>0.05）.The AA genotype of TNF-α 308 locus was not detected in the study.Compared with the control group［17 cases（10.6%）］，the distribution proportion of GA genotype in the CFS group［22cases（25.0%）］and the SFS group［52cases（22.4%）］was increased，and the difference was statistically significant（ χ2＝11.126， P＝0.004）；Compared with the control group［17 frequencies（5.3%）］，the frequency distribution proportion of allele A in the CFS group［22 frequencies（12.5%）］and SFS group［52 frequencies（11.2%）］was also increased，and the difference was statistically significant（ χ2＝9.960， P＝0.007）. Adding control factors such as gender，age，family history of FS，body temperature at time of convulsions and blood routine markers，the multivariate Logistic regression model was constructed to show that there was no statistically significant association between TNF-α 308 genotype and CFS in children（ OR＝1.805，95% CI：0.926~3.519， P＝0.083）. Conclusion:In this study，there was no significant correlation between TNF-α 308 gene loci polymorphism and CFS in children.

Objective To investigate the prevalence and genotypes of Cryptosporidium spp. and Giardia lamblia in dogs and cats from a pet hospital in Shanghai Municipality. Methods A total of 145 fresh fecal samples were collected from pet dogs and cats in a pet hospital in Shanghai Municipality during the period from November 2021 to June 2022, including 99 dog fecal samples and 46 cat fecal samples. The small subunit ribosomal ribonucleic acid (SSU rRNA) gene of Cryptosporidium and the triose phosphate isomerase (TPI) gene of G. lamblia were amplified using nested PCR assay, and the positive amplification products were sequenced from both directions. The sequence assembly was performed using the software Clustal X 2.1, and sequence alignment was conducted using BLAST. A phylogenetic tree was created with the Neighbor-Joining method using MEGA 11.0 to identify parasite species or genotype. Results The overall prevalence of Cryptosporidium and G. lamblia was 20.00% (29/145) in 145 pet dog and cat fecal samples, with the prevalence of 0.69% (1/145) and 19.31% (28/145) in Cryptosporidium and G. lamblia, respectively. G. lamblia was only detected in dog fecal samples, with prevalence of 18.18% (18/99), while the detection rates of Cryptosporidium and G. lamblia were 2.17% (1/46) and 21.74% (10/46) in cat fecal samples. Nucleotide sequence analysis showed that one Cryptosporidium positive sample was characterized as C. felis, and 28 G. lamblia positive samples were all characterized as Giardia assemblage A, which showed 100% sequence homology with human isolates of Giardia. Phylogenetic analysis revealed that the sequences obtained in this study belonged to the same branch with the reported Giardia assemblage A. Conclusions Cryptosporidium and G. lamblia infection was prevalent in pet dogs and cats from the study pet hospital in Shanghai Municipality, and there is a zoonotic risk for the species and genotype. Intensified surveillance of Cryptosporidium and Giardia infection is recommended in pets and their owners, and improved management of pet keeping is required.

Objective:To analyze the independent diagnostic indicators and their diagnostic values for pulmonary tuberculosis complicated with pulmonary embolism.Methods:A total of 34 cases of pulmonary tuberculosis complicated with pulmonary embolism treated in Huzhou Central Hospital from March 2014 to September 2021 were enrolled. And 136 patients with simple pulmonary tuberculosis who were hospitalized during the same period were collected with a ratio of 1∶4 according to the principle of age and gender matching. The general conditions, clinical symptoms, comorbidities and laboratory indicators of the patients were retrospectively analyzed. The univariate analysis was performed using independent samples t test, Mann-Whitney U test and chi-square test. Binary logistic regression was used to analyze the related diagnostic factors for pulmonary embolism in pulmonary tuberculosis patients, and the combined factors were constructed by transforming the model equation, and the receiver operating characteristic (ROC) curve was used to determine the optimal cut-off value and evaluate its diagnostic value. Results:The univariate analysis showed that patients with pulmonary tuberculosis complicated with pulmonary embolism had higher ratio of chest tightness (67.6%(23/34) vs 22.1%(30/136)), syncope (23.5%(8/34) vs 0.7%(1/136)), fever (55.9%(19/34) vs 36.0%(49/136)), hemostatic drug use (100.0%(34/34) vs 13.2%(18/136)), history of venous thrombosis (8.8%(3/34) vs 0.7%(1/136)), atrial fibrillation (11.8%(4/34) vs 2.2%(3/136)) and D-dimer levels (4.090 0(1.035 0, 10.790 0) mg/L vs 0.850 0(0.432 5, 2.145 0) mg/L) than those of simple pulmonary tuberculosis patients, and the differences were all statistically significant ( χ2=26.35, 28.19, 4.47, 96.44, 7.75, 6.30 and Z=-4.65, respectively; all P<0.050). The arterial partial pressure of oxygen (PaO 2)(61.90(52.95, 73.00) mmHg vs 82.00 (75.00, 87.00) mmHg, 1 mmHg=0.133 kPa) and albumin ((28.83±4.98) g/L vs (32.76±5.65) g/L) of patients with pulmonary tuberculosis complicated with pulmonary embolism were lower than those of simple pulmonary tuberculosis patients, and the differences were both statistically significant ( Z=-5.21 and t=3.71, respectively, both P<0.001). Binary regression analysis showed that chest tightness (odds ratio ( OR)=3.494, 95%confidence interval ( CI) 1.208 to 10.100, P=0.021), D-dimer ( OR=1.285, 95% CI 1.079 to 1.530, P=0.005) and PaO 2( OR=0.931, 95% CI 0.895 to 0.970, P=0.001) were the independent diagnostic indicators for pulmonary embolism in pulmonary tuberculosis patients. The areas under the ROC curve of chest tightness, D-dimer, PaO 2, and the combination of the three indicators (the combination factor) were 0.728, 0.758, 0.834, and 0.890, respectively. The optimal cut-off value of the combination factor was -3.1, with the sensitivity of 0.824 and the specificity of 0.824. Conclusions:Chest tightness, increased D-dimer and decreased PaO 2 are independent diagnostic indicators for pulmonary embolism in pulmonary tuberculosis patients. It is recommended to perform pulmonary artery computed tomography angiography promptly when the combination factor is higher than -3.1 to determine whether the patient is complicated by pulmonary embolism.

Objective:To analyze the causes, influencing factors and trends of dead on arrival cases in children′s Hospital in the past 5 years, aiming to provide direction and basis for reducing the dead on arrival cases of children.Methods:We collected the dead on arrival cases in the department of emergency at Shanghai Children′s Hospital from January 2015 to December 2019, classifed and analysed the gender, age, native place, death season, time of death, and possible causes of death, and then studied the correlation between above factors and the cases.Results:A total of 151 dead on arrival cases were collected.The annual number decreased year by year, and boys were more than girls in gender.Most of them were infants under 1 year old, and nonlocal children were more than Shanghai native.The above differences were statistically significant, but there was no significant difference in the distribution of death season and death time.In terms of the cause of death, perinatal diseases accounting for 33.8%(51/151), those accompanied with severe underlying diseases accounting for 39.1%(59/151), accidental death accounting for 14.6%(22/151), unexplained deaths accounting for 12.6%(19/151). Those distribution differences were statistically significant( χ2=32.497, P<0.001). Meanwhile, there were statistic differences in gender and age of the cases with severe underlying diseases( χ2=4.898, P=0.027; χ2=32.169, P<0.001), and the year and age distributions of the accidental death cases also had significant differences( χ2=16.636, P=0.002; χ2=14.727, P=0.002). Conclusion:To reduce dead on arrival cases of children, we should do a good job in perinatal health care and screening, reduce premature birth and birth defects, actively conduct propaganda to prevent children′s accidental injuries, popularize medical first aid knowledge, and strengthen children′s transport system.

Objective:To analyze the clinical characteristics of brucellosis in Huzhou City, Zhejiang Province, so as to provide basis for clinicians to improve their understanding and diagnosis and treatment of brucellosis.Methods:The data of 10 cases of brucellosis in Huzhou Central Hospital from 2014 to 2019 were collected, and the epidemiological characteristics, clinical manifestations, laboratory examination results, imaging examination results, etiological examination results, diagnosis and treatment process and prognosis were summarized and analyzed by retrospective analysis method.Results:Among the 10 patients with brucellosis, 6 were males and 4 were females, aged from 32 to 71 years old, and 9 patients had a history of contact with goat, and one case was considered as food borne infection. The main clinical manifestations were fever (10 cases), sweating (2 cases), joint pain (7 cases). In imaging examination, 5 cases with splemomegaly, 1 cases with lymphadenopathy, 3 patients presented with spondylitis (including 1 case with paravertebral abscess) and 2 cases with prostatitis; Brucella was cultured in 2 patients and Brucella antibodies were positive in 10 patients. The treatment drugs included doxycycline, rifampicin and levofloxacin in two or three combinations. Eight patients were followed up for more than 1 month to more than 5 years, and the prognosis was good. Conclusions:Brucellosis lacks specificity in clinical features. Most patients with brucellosis are associated with osteoarthritis or urinary system diseases. Physicians should be vigilant, combine epidemiological features, conduct laboratory tests in time, and diagnose as soon as possible.

Objective To establish the model of rabbit vena arterialization, so as to investigate the difference of mechanical parameters between arteries and veins as well as before and after arterialization. Methods Twenty-four rabbits were randomly divided into experimental group (n=12) and control group (n=12). By establishing the rabbit vena arterialization model for experimental group, the arterial blood could flow into the veins. After model creation, the vein would be removed 4 weeks after surgery. In the meantime, the external jugular veins and cephalic arteries extracted from control group were acquired. Compressive pressurizing and stretching tests on all vessels were conducted at the same time(including arteries, veins and arterialized veins). Observation was supported by HE staining and immune tissue chemical techniques. Results There were no deaths among the 24 rabbits, with unobstructed blood flow in veins. With the increase of intravascular pressure, the outer diameter of veins changed at first and then stabilized at a fixed value. The elasticity of veins was worse than that of arteries. The external diameter of veins increased rapidly with internal pressure of veins increasing and reached its extreme elasticity. Comparatively, the elasticity of arteries increased slowly. HE staining results showed that thickness of the vascular wall was thinner, while it became thicker after vena arterialization. After vena arterialization, proliferating cell nuclear antigen（PCNA） and α-actin showed positive results. It further proved that proliferation existed among smooth muscle cells, and veins showed the tendency of restenosis again. The elasticity of veins after transplantation into the arterial system was improved compared with that before transplantation. Conclusions Accompanied by the increasing pressure, the vein could reach its elasticity extremity faster than the artery. Under such a long-term high pressure, vein intima was vulnerable. After vena arterialization, with the gradual thickening of vein intima, the tendency of vessel restenosis was obvious, and the elasticity of veins has been improved after transplantation.