The standard therapy bundle for septic shock includes antibiotics,fluid resuscitation,vasopressors and inotropes. Given the high mortality rates associated with septic shock,there is an ongoing need for active research into potential therapies to improve outcomes. Venoarterial extracorporeal membrane oxygenation (VA-ECMO) is an acceptable and beneficial therapeutic modality for patients who fail conventional management and have decreased left ventricular ejection fraction. Up to now,the benefits of ECMO in pediatric sepsis remain unclear. Whether ECMO is the rescue therapy or is appropriately utilized in the early stage (window forward) remains controversial. At the present stage,it is necessary to strictly screen patients and make individualized decisions based on dynamic hemodynamic monitoring and biomarkers.

Objective:To explore the clinical features and risk factors for pediatric severe Mycoplasma pneumoniae pneumonia (SMPP) complicated with pulmonary embolism. Methods:SMPP patients admitted to Department of Pediatrics, Jiaxing First Hospital and Pediatric Intensive Care Unit, Shanghai Children′s Hospital from December 2019 to December 2023 were included in this retrospective case-control study.According to whether they were complicated with pulmonary embolism, SMPP patients were divided into a pulmonary embolism group and a non-pulmonary embolism group.Clinical characteristics of the two groups, including general data, laboratory examination and imaging data were compared and analyzed.The t-test and Mann-Whitney rank-sum test were used to compare the measurement data, and the χ2 test was used to compare the count data.The risk factors of SMPP patients developing pulmonary embolism were analyzed by the univariate method. Results:There were 10 out of 62 SMPP children developing pulmonary embolism, showing an incidence of 16.13%.In the pulmonary embolism group, there were 5 boys and 5 girls, with a median age of 7.50 (5.75, 9.25) years.There were 52 children in the non-pulmonary embolism group, including 29 boys and 23 girls, with a median age of 6.50(5.00, 8.00)years.The hospitalization time, body temperature, total white blood cell count, neutrophil count, C-reactive protein levels, lactate dehydrogenase levels, prothrombin time, activated partial thromboplastin time, D-dimer (D-D) levels, fibrinogen degradation product levels, pleural effusion and atelectasis rates in the pulmonary embolism group were significantly higher than those in the non-pulmonary embolism group (all P<0.05). Fibrinogen levels in the pulmonary embolism group were significantly lower than those in the non-pulmonary embolism group ( P<0.05). Univariate Logistic regression analysis showed that the D-D level was a risk factor for SMPP patient developing pulmonary embolism.The receiver operating characteristic curve analysis revealed that the D-D level had the largest area under the curve for predicting pulmonary embolism of 0.990(95% CI: 0.972-1.000, P<0.001), with a sensitivity of 100%, a specificity of 92%, and a cut-off value of 4.63 mg/L. Conclusions:SMPP children complicated with pulmonary embolism are prone to high inflammation and impaired coagulation function.The increase of D-D levels is a risk factor for the development of pulmonary embolism in SMPP.

Objective:To investigate the role and mechanism of negative pressure wound therapy (NPWT) in a rabbit full-thickness wound model using non-targeted metabolomics.Methods:Eighteen male New Zealand rabbits (11-12 weeks old) were used. Two symmetrical circular full-thickness skin defects were created on the back of each rabbit. The animals were randomly divided into three groups: Control group (no treatment), Saline group (debridement with saline irrigation), and NPWT+ Saline group (saline debridement followed by 2 h of NPWT at -125 mm Hg once daily for two weeks). Wound healing was documented on days 0, 3, 7, 10, and 14. The wound healing rate was calculated as (original area-unhealed area)/original area × 100%. Histopathological changes were evaluated via hematoxylin and eosin (HE) staining. Metabolomic profiling of wound tissues was performed using ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Differential metabolites were identified, and pathway enrichment analysis was conducted. Oxidative stress markers, including superoxide dismutase (SOD) and catalase (CAT) activities and malondialdehyde (MDA) content, were measured using commercial kits. Data were analyzed using SPSS 20.0. One-way ANOVA with Tukey’s HSD test or Welch’s ANOVA with Games-Howell test was applied as appropriate.Results:On days 3, 10, and 14, the wound healing rate in the NPWT+ Saline group was significantly higher than that in the Control and Saline groups ( P<0.05). On day 7, the NPWT+ Saline group showed a significantly higher healing rate than the Saline group ( P<0.01), but no significant difference compared with the Control group ( P>0.05). HE staining on day 7 revealed enhanced epithelialization, thicker granulation tissue, higher microvessel density, and more abundant, well-organized collagen in the NPWT+ Saline group. By day 14, all groups had formed relatively continuous epithelial structures. Non-targeted metabolomics identified riboflavin and spermidine as differential metabolites. Pathway analysis highlighted riboflavin metabolism and glutathione metabolism as the most significantly enriched pathways. Compared with the Control and Saline groups, the NPWT+ Saline group exhibited significantly increased CAT and SOD activities ( P<0.05) and decreased MDA content ( P<0.01), indicating reduced oxidative stress. Conclusion:NPWT may promote wound healing by elevating riboflavin and spermidine levels, thereby modulating riboflavin and glutathione metabolism and regulating local redox reactions.

Objective:To investigate the role and mechanism of negative pressure wound therapy (NPWT) in a rabbit full-thickness wound model using non-targeted metabolomics.Methods:Eighteen male New Zealand rabbits (11-12 weeks old) were used. Two symmetrical circular full-thickness skin defects were created on the back of each rabbit. The animals were randomly divided into three groups: Control group (no treatment), Saline group (debridement with saline irrigation), and NPWT+ Saline group (saline debridement followed by 2 h of NPWT at -125 mm Hg once daily for two weeks). Wound healing was documented on days 0, 3, 7, 10, and 14. The wound healing rate was calculated as (original area-unhealed area)/original area × 100%. Histopathological changes were evaluated via hematoxylin and eosin (HE) staining. Metabolomic profiling of wound tissues was performed using ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Differential metabolites were identified, and pathway enrichment analysis was conducted. Oxidative stress markers, including superoxide dismutase (SOD) and catalase (CAT) activities and malondialdehyde (MDA) content, were measured using commercial kits. Data were analyzed using SPSS 20.0. One-way ANOVA with Tukey’s HSD test or Welch’s ANOVA with Games-Howell test was applied as appropriate.Results:On days 3, 10, and 14, the wound healing rate in the NPWT+ Saline group was significantly higher than that in the Control and Saline groups ( P<0.05). On day 7, the NPWT+ Saline group showed a significantly higher healing rate than the Saline group ( P<0.01), but no significant difference compared with the Control group ( P>0.05). HE staining on day 7 revealed enhanced epithelialization, thicker granulation tissue, higher microvessel density, and more abundant, well-organized collagen in the NPWT+ Saline group. By day 14, all groups had formed relatively continuous epithelial structures. Non-targeted metabolomics identified riboflavin and spermidine as differential metabolites. Pathway analysis highlighted riboflavin metabolism and glutathione metabolism as the most significantly enriched pathways. Compared with the Control and Saline groups, the NPWT+ Saline group exhibited significantly increased CAT and SOD activities ( P<0.05) and decreased MDA content ( P<0.01), indicating reduced oxidative stress. Conclusion:NPWT may promote wound healing by elevating riboflavin and spermidine levels, thereby modulating riboflavin and glutathione metabolism and regulating local redox reactions.

The standard therapy bundle for septic shock includes antibiotics,fluid resuscitation,vasopressors and inotropes. Given the high mortality rates associated with septic shock,there is an ongoing need for active research into potential therapies to improve outcomes. Venoarterial extracorporeal membrane oxygenation (VA-ECMO) is an acceptable and beneficial therapeutic modality for patients who fail conventional management and have decreased left ventricular ejection fraction. Up to now,the benefits of ECMO in pediatric sepsis remain unclear. Whether ECMO is the rescue therapy or is appropriately utilized in the early stage (window forward) remains controversial. At the present stage,it is necessary to strictly screen patients and make individualized decisions based on dynamic hemodynamic monitoring and biomarkers.

Objective:To explore the clinical features and risk factors for pediatric severe Mycoplasma pneumoniae pneumonia (SMPP) complicated with pulmonary embolism. Methods:SMPP patients admitted to Department of Pediatrics, Jiaxing First Hospital and Pediatric Intensive Care Unit, Shanghai Children′s Hospital from December 2019 to December 2023 were included in this retrospective case-control study.According to whether they were complicated with pulmonary embolism, SMPP patients were divided into a pulmonary embolism group and a non-pulmonary embolism group.Clinical characteristics of the two groups, including general data, laboratory examination and imaging data were compared and analyzed.The t-test and Mann-Whitney rank-sum test were used to compare the measurement data, and the χ2 test was used to compare the count data.The risk factors of SMPP patients developing pulmonary embolism were analyzed by the univariate method. Results:There were 10 out of 62 SMPP children developing pulmonary embolism, showing an incidence of 16.13%.In the pulmonary embolism group, there were 5 boys and 5 girls, with a median age of 7.50 (5.75, 9.25) years.There were 52 children in the non-pulmonary embolism group, including 29 boys and 23 girls, with a median age of 6.50(5.00, 8.00)years.The hospitalization time, body temperature, total white blood cell count, neutrophil count, C-reactive protein levels, lactate dehydrogenase levels, prothrombin time, activated partial thromboplastin time, D-dimer (D-D) levels, fibrinogen degradation product levels, pleural effusion and atelectasis rates in the pulmonary embolism group were significantly higher than those in the non-pulmonary embolism group (all P<0.05). Fibrinogen levels in the pulmonary embolism group were significantly lower than those in the non-pulmonary embolism group ( P<0.05). Univariate Logistic regression analysis showed that the D-D level was a risk factor for SMPP patient developing pulmonary embolism.The receiver operating characteristic curve analysis revealed that the D-D level had the largest area under the curve for predicting pulmonary embolism of 0.990(95% CI: 0.972-1.000, P<0.001), with a sensitivity of 100%, a specificity of 92%, and a cut-off value of 4.63 mg/L. Conclusions:SMPP children complicated with pulmonary embolism are prone to high inflammation and impaired coagulation function.The increase of D-D levels is a risk factor for the development of pulmonary embolism in SMPP.

Objective:To investigate the role and related mechanisms of the LiaSR two-component system in acid tolerance and biofilm formation abilities of Streptococcus mutans (Sm) 593. Methods:The growth curves of various Sm strains in pH=5.5 brian heart infusion (BHI) medium were analyzed. And colony forming unit (CFU) was also performed to evaluate the acid tolerance of Sm. Laurdan probe, H +-K +adenosine triphosphate (ATP)ase activity analysis kit, proton permeability assay and real-time fluorescence quantitative PCR (RT-qPCR) were conducted to detect the acid tolerant mechanisms of LiaSR two-component system in Sm. Crystal violet staining, CFU, SYTOX probe and anthrone-sulfuric method were used to analyze the properties and structures of the Sm biofilms. RT-qPCR was conducted to detect the expression levels of underlying regulated genes. Results:The growth of mutants in acidic BHI were inhibited ( P<0.05). The acid tolerance of mutants significantly decreased compared to the wild-type strain ( P<0.05). In mutants, the activity of H +-ATPase (917.06±59.53 and 469.53±47.65) were elevated by 7.22-folds and 3.70-folds compared to the wild-type strain (127.00±50.71) ( P<0.001, P<0.001) and the encoded gene atpD (3.39±0.21 and 1.94±0.17) were also elevated by 3.39-folds and 1.94-folds compared to the wild-type strain (1.00±0.15) ( P<0.001, P=0.001). The Laurdan generalized polarization of mutants (0.18±0.04 and 0.18±0.05) increased significantly compared to the wild-type strain (0.08±0.05) ( P=0.006, P=0.003) and the expression levels of fabM gene were decreased in mutants (0.52±0.11 and 0.57±0.05) by 1/2 ( P=0.014, P=0.022). In liaR deletion mutant, the reduced terminal pH (4.76±0.01) can also be observed ( P<0.001). The total amount of the biofilms of three Sm didn't show significant differences ( P>0.05). But the number of viable bacteria of mutants′ biofilms were decreased [Sm 593: (12.00±2.80)×10 7 CFU/ml; Sm ΔliaS: (2.95±1.13)×10 7 CFU/ml; Sm ΔliaR: (7.25±1.60)×10 7 CFU/ml] ( P=0.001, P=0.024). The extracellular DNA were increased by 18.00-folds and 6.50-folds in mutants′ biofilms (128.73±15.65 and 46.38±5.52) compared to the wild-type strain (7.16±3.62) ( P<0.001, P=0.003). Water-soluble exopolysaccharides could be found up-regulated in liaS deletion mutant [(138.73±10.12) μg/ml] ( P=0.003) along with the expression level of gtfC gene (1.65±0.39) ( P=0.014). The expression level of gtfD were elevated by 47.43-folds and 16.90-folds in mutants ( P<0.001, P=0.010). Conclusions:The LiaSR two-component system can promote the expression of fabM gene and increase the fluidity of Sm which contributes to acid tolerance. The LiaR can also decrease the proton permeability and restrict the entrance of H +. The LiaSR two-component system can negatively regulate the production of the extracellular matrix in Sm biofilm.

Objective:To evaluate the feasibility of using positive anti-M-type phospholipase A 2 receptor(PLA 2R)antibody detection as a surrogate for renal biopsy in the diagnosis of idiopathic membranous nephropathy(IMN)in elderly Chinese. Methods:From June 2021 to March 2022, clinicopathological data of ninety-six elderly patients(≥60 years old)with positive anti-PLA 2R antibody detection(ELISA≥14 RU/ml), complete renal pathology records, and the exclusion of secondary disease and diabetes mellitus were collected from the Department of Nephrology, The First Affiliated Hospital of Zhengzhou University.Patients were divided into high eGFR group(≥60 ml·min -1·1.73 m -2)and low eGFR group(<60 ml·min -1·1.73 m -2), and the data were analyzed retrospectively. Results:Of the 96 patients, 95(99.0%)had IMN(1 in stage Ⅰ, 59 in stage Ⅱ, 34 in stage Ⅲ, and 1 in stage Ⅳ), and 1(1.0%)had atypical membranous nephropathy(AMN).For the IMN patients, 81(85.3%)had high eGFR and most IMN patients(66.7%)were in stage Ⅱ, with 10(12.3%)showing additional pathological findings, including mild mesangial proliferative IgA nephropathy(2 cases)and subacute tubulointerstitial nephropathy(2 cases).There were 14 patients(14.7%)with low eGFR, and most of them(57.1%)had stage Ⅲ IMN, with 10(71.4%)showing additional pathological findings.This percentage was higher than that in the high eGFR group( χ2=21.642, P<0.05).The most common additional pathological findings were acute tubular injury(4 cases)and ischemic kidney injury(2 cases). Conclusions:Positive anti-PLA 2R antibody detection is highly predictive of IMN in elderly Chinese patients, but it often co-exists with other pathological entities.The advantages of renal biopsy in detecting other pathological conditions and the risks associated with the procedure should be evaluated.

Objective:To analyze the clinical features of severe refractory mycoplasma pneumoniae pneumonia (SRMPP) in children, and explore its risk factors complicated with extrapulmonary organ dysfunction.Methods:The clinical data of children with SRMPP who were admitted to the Department of Critical Care Medicine of Shanghai Children's Hospital from July 2017 to June 2019 were retrospectively summarized. The patients were divided into two groups according to the occurrence of extrapulmonary organ dysfunction: the extrapulmonary organ dysfunction group and the respiratory dysfunction group. The differences of clinical features and laboratory indexes between the two groups were compared, and the risk factors of extrapulmonary organ dysfunction were screened out by logistic regression analysis.Results:A total of 107 cases with SRMPP were admitted to the Pediatric Intensive Care Unit during the past two years, and there were 44 cases (41.1%) complicated with pleural effusion, 17 cases (15.9%) with plastic bronchitis, 104 cases (97.2%) with positive results for macrolide resistance genes (2063, 2064), with an in-hospital mortality rate of 2.8% (3/107). Among 107 children with SRMPP, there were 51 cases (47.7%) with extrapulmonary organ dysfunction, 43 cases (40.2%) with cardiovascular dysfunction, 13 cases (12.1%) with coagulation dysfunction, 11 cases (10.3%) with gastrointestinal dysfunction, 4 cases (3.7%) with renal dysfunction, 4 cases (3.7%) with brain dysfunction, 3 cases (2.8%) with liver dysfunction, and 16 cases (15.0%) with multiple organ dysfunction. Compared with the respiratory dysfunction group, the incidence of capillary leak syndrome was higher (52.9% vs. 17.9%, P < 0.001), the capillary leak index was increased [11.71 (4.63, 27.07) vs. 5.78 (2.07, 15.71), P =0.019], serum albumin was decreased [(32.2 ± 5.6)g/L vs. (34.7 ± 6.7)g/L, P=0. 041], and prothrombin time was prolonged significantly [12.7 (11.7, 13.8)s vs. 12.0 (11.4, 13.0)s, P=0. 009]. Logistic regression analysis showed that capillary leak syndrome ( OR=0. 278, 95% CI 0.102-0.759, P=0. 013) and prolonged prothrombin time ( OR=1. 443, 95% CI 1.018-2.046, P=0. 039) were independent risk factors for SRMPP complicated with extrapulmonary organ dysfunction. Conclusions:Approximately 50% of children with SRMPP have dysfunction of extrapulmonary organs, such as circulation, coagulation and gastrointestinal disorders. Capillary leak syndrome and prolonged prothrombin time are independent risk factors for SRMPP complicated with extrapulmonary organ dysfunction.

Objective:To evaluate the predictive value of lung ultrasound on mortality in children with severe acute respiratory distress syndrome (ARDS) undergoing extracorporeal membrane oxygenation (ECMO) support.Methods:A prospective observational study was used to enroll patients with severe ARDS who met the Berlin criteria in the Pediatric Intensive Care Unit of Children’s Hospital of Shanghai Jiao Tong University from January 2016 to December 2019. Patients with ECMO support <3 d, lack of appropriate acoustic windows, with severe pneumothorax, and secondary to congenital heart disease or chronic lung disease were excluded. ECMO parameters, respiratory mechanics parameters and outcome were collected and analyzed. Lung ultrasound score (LUS) was measured at the initiation of ECMO as LUS-0 h, then at 24 h, 48 h, 72 h, and 7 d after ECMO support as the value of LUS-24 h, LUS-48 h, LUS-72 h, LUS-7 d, as well as after weaning ECMO as LUS-w. The patients were divided into survivors and non-survivors according to hospital survival status. Receiver operating characteristic (ROC) curve and Kaplan-Meier survival analysis curve were performed to explore the predictive value of lung ultrasound on mortality in patients with severe ARDS undergoing ECMO.Results:A total of 26 patients were enrolled in this study, of which 18 patients survived and 8 died. There were no significant differences in PRISM Ⅲ, dynamic pulmonary compliance (Cdyn), oxygenation index, PaO 2/FiO 2, and PaCO 2 on PICU admission between the two groups (all P>0.05). The values of LUS-72 h and LUS-w in non-survivors were significantly higher than those in survivors [26 (24, 29) vs16 (13, 19), P<0.01] and [30 (26, 35) vs11 (10, 13), P<0.01]. The values of Cdyn-72 h, Cdyn-7 d and Cdyn-w in survivors were significantly higher than those in non-survivors [0.48 (0.42, 0.54)mL/cmH 2O·kg vs 0.36 (0.29, 0.40) mL/cmH 2O·kg, P<0.01; 0.60 (0.52, 0.67) mL/cmH 2O·kg vs 0.27 (0.13, 0.30) mL/cmH 2O·kg, P<0.01, and 0.66 (0.62, 0.70) mL/cmH 2O·kg vs 0.30 (0.13, 0.35) mL/cmH 2O·kg, P<0.01]. ROC curve analysis showed that an area under ROC curve (AUC) of LUS-72 h for predicting PICU mortality was 0.955 (95% CI: 0.864-1.000; P<0.01). The cutoff value of LUS-72 h was 24 with a sensitivity of 87.5% and a specificity of 100.0%. Kaplan-Meier survival analysis showed that PICU mortality of patients with LUS-72 h≥24 was significantly higher than that in patients with LUS-72 h < 24 ( P<0.01) . Conclusions:Lung ultrasound is an effective tool for monitoring progress of children with severe ARDS received ECMO support. LUS-72 h >24 is an index to predict the worsen outcome in children with severe ARDS under ECMO support.