PRKCA Promotes Mitophagy through the miR-15a-5p/PDK4 Axis to Relieve Sepsis-Induced Acute Lung Injury.

Acute lung injury (ALI) caused by sepsis is a common respiratory critical illness with high morbidity and mortality. Protein kinase C-alpha (PRKCA) plays a protective role in sepsis-induced ALI. However, the detailed molecular mechanism of PRKCA in ALI caused by sepsis is unclear. Animal and cell models of sepsis were established by cecal ligation and puncture (CLP)-surgery and lipopolysaccharide (LPS)/interferon-gamma (IFN-γ) treatment, respectively. Lentivirus transfection was used to overexpress PRKCA. H&E staining and lung injury in CLP-surgery mice were evaluated. Gene expression was evaluated using qPCR and Western blotting. The expression of TNF-α, IL-1ß, and IL-6 was examined using qPCR and ELISA. The expression of LC3 and TOM20 was evaluated using immunofluorescence assays. Cell apoptosis was assessed using a flow cytometry assay. The bond between miR-15a-5p and PDK4 was confirmed by dual-luciferase reporter gene and RNA immunoprecipitation assays. In vivo and in vitro, PRKCA overexpression reduced lung injury to prompt mitophagy and inhibit the inflammatory response, ROS production, and cell apoptosis. miR-15a-5p was highly expressed in macrophages treated with LPS/IFN-γ and was negatively mediated by PRKCA. The overexpression of miR-15a-5p reduced the effects of PRKCA upregulation in macrophages. miR-15a-5p could restrain mitophagy in LPS/IFN-γ-treated macrophages by directly targeting PDK4. Furthermore, PDK4 knockdown reversed the inhibition of cell apoptosis and inflammatory factor release caused by miR-15a-5p silencing. The PRKCA/miR-15a-5p/PDK4 axis alleviated ALI caused by sepsis by promoting mitophagy and repressing anti-inflammatory response.

Development of a Chinese Version of the State Behavioral Scale for Mechanically Ventilated Children.

PURPOSE: To develop a Chinese version of the State Behavioral Scale (SBS-C) and to evaluate its reliability and validity for sedation assessment in mechanically ventilated children in China. DESIGN AND METHODS: Cross-sectional survey design was used in a two-part study of mechanically ventilated children, aged 6 weeks to 6 years. A total 172 children and 145 children were recruited from Jan-Dec 2017 and Jan-Dec 2018, respectively, at a tertiary care pediatric hospital in southeast China. Following translation of the scale, the content validity was established by the content validity index, internal consistency was established using Cronbach's α, and construct validity was confirmed by correlation with a similar well-recognized scale, the COMFORT Scale-Chinese version (CS-C). RESULTS: The content validity index for the seven scale dimensions ranged from 0.83 to 1.0 and for the full scale was 0.932. In the first study, Cronbach's α for the full SBS-C was 0.986 and for the seven scale dimensions ranged from 0.973 to 0.983; similarly, in the second study, Cronbach's α for the full scale was 0.983 and for the seven dimensions ranged from 0.977 to 0.987. The correlation coefficient between scores of the SBS-C and the CS-C was 0.919 (P < .01). CONCLUSIONS: The SBS-C is valid, reliable, and responsive and is suitable for assessing sedation in mechanically ventilated children in China. IMPLICATIONS FOR PRACTICE: The SBS-C can be used for sedation assessment in mechanically ventilated children in China, guiding decision making and the provision of care, and optimizing patient safety.

Epstein-Barr virus-related hemophagocytic lymphohistiocytosis complicated with coronary artery dilation and acute renal injury in a boy with a novel X-linked inhibitor of apoptosis protein (XIAP) variant: a case report.

BACKGROUND: X-linked lymphoproliferative disease (XLP) is a rare inherited X-linked primary immunodeficiency diseases (PID). One such disease, X-linked inhibitor of apoptosis protein (XIAP) deficiency, is characterized by Epstein-Barr virus-related hemophagocytic lymphohistiocytosis (EBV-HLH). However, EBV-HLH with coronary artery dilation and acute renal injury (AKI) in children is unusual. CASE PRESENTATION: We report the case of a young boy aged 17 months with a novel XIAP variant. He was initially diagnosed with EBV-HLH based on the HLH-2004 diagnostic criteria and the condition was accompanied by coronary artery dilation and acute renal injury. The comprehensive genetic analysis of peripheral blood-derived DNA revealed a hemizygous variant of the XIAP gene [c.116G > C(p.G39A)], which was inherited from his mother (heterozygous condition). After combined treatment with rituximab, intravenous immunoglobulin, corticosteroids, antiviral drugs, and mycophenolate mofetil (MMF) in addition to supportive therapy, his clinical manifestations and laboratory indexes were improved. The patient achieved complete remission with MMF treatment in the 8-month follow-up. CONCLUSIONS: We report the [c.116G > C(p.G39A)] variant in the XIAP gene for the first time in a case of XLP-2 associated with EBV-HLH. For male patients with severe EBV-HLH, the possibility of XLP should be considered and molecular genetic testing should be used early in auxiliary diagnosis. Reports of EBV-HLH with coronary artery dilation and AKI in children are rare. In the patients with EBV-HLH, color Doppler echocardiography and urine tests should be monitored regularly. If necessary, renal biopsy can be performed to clarify the pathology. Treatment with rituximab, immunosuppressors and supportive therapy achieved a good effect, but long-term follow-up is required.

X-Linked Hyper IgM Syndrome Manifesting as Recurrent Pneumocystis jirovecii Pneumonia: A Case Report.

We reported a Chinese boy with X-linked hyper IgM (XHIGM) syndrome, manifesting as recurrent and severe pneumonia caused by Pneumocystis jirovecii. His parents were healthy and unrelated. In August 2018, the 5-month-old boy manifested as cough and dyspnea, and then in July 2019, he was admitted because of the same symptoms. Immunological results of the two admissions both showed low IgG, low IgA, normal IgM and high levels of 1,3-ß-D-glucan (BDG). Using next-generation sequencing (NGS), great reading counts of P. jirovecii were identified from the deep sputum in both admissions. Caspofungin combined with trimethoprim-sulfamethoxazole were used to anti-infection, and he recovered quickly. Whole-exome sequencing was performed for this family because of immune suppression, the disease-causing gene (exon 10-22 of CD40L) deletion for XHIGM syndrome was identified. NGS is beneficial for etiology diagnosis. Pneumocystis jirovecii pneumonia as an opportunistic infection could be recurrent in patients with XHIGM syndrome.

[Value of amplitude-integrated EEG combined with Full Outline of Unresponsiveness in evaluating the prognosis of children with disturbance of consciousness].

OBJECTIVE: To study the value of amplitude-integrated EEG (aEEG), Full Outline of Unresponsiveness (FOUR), and Glasgow Coma Scale (GCS) in evaluating the prognosis of children with disturbance of consciousness in the pediatric intensive care unit (PICU). METHODS: A total of 164 children with disturbance of consciousness who were admitted to the PICU of Children's Hospital Affiliated to Soochow University were enrolled as subjects. According to prognosis, they were divided into a poor prognosis group with 111 children and a good prognosis group with 53 children. The results of aEEG monitoring, FOUR score, and GCS score on days 1 and 5 of admission were collected. The association between evaluation methods and prognosis was analyzed. The receiver operating characteristic (ROC) curve was used to evaluate the value of aEEG, FOUR, and GCS in predicting prognosis. RESULTS: The children with no improvement or abnormal aggravation of aEEG on day 5 tended to have a poor prognosis. The results of aEEG was positively correlated with prognosis (r=0.689, P<0.001), and FOUR and GCS were negatively correlated with prognosis (r=-0.655 and -0.554 respectively, P<0.001). The areas under the ROC curve (AUC) of aEEG, FOUR, and GCS were 0.894, 0.903, and 0.840 respectively, and there was no significant difference in the AUC between the three indices (P>0.05), while aEEG combined with FOUR had an AUC of 0.945, which was significantly larger than that of each index alone (P<0.05). CONCLUSIONS: Both aEEG and FOUR can be used as effective tools to predict the prognosis of children with disturbance of consciousness, and a combination of aEEG and FOUR can improve the predictive value.

[Value of three scoring systems in evaluating the prognosis of children with severe sepsis].

OBJECTIVE: To study the predictive value of Pediatric Age-adapted Sequential Organ Failure Assessment Score (pSOFA), Pediatric Risk of Mortality Score III (PRISM III), and Pediatric Critical Illness Score (PCIS) in children with severe sepsis. METHODS: A retrospective analysis was performed for the clinical data of 193 hospitalized children with severe sepsis. According to the final outcome, these children were divided into a survival group with 151 children and a death group with 42 children. The scores of pSOFA, PRISM III, and PCIS were determined according to the worst values of each index within 24 hours after admission. The receiver operating characteristic (ROC) curve was used to analyze the efficiency of each scoring system in predicting the risk of death due to sepsis. Smooth curve fitting was used to analyze the correlation between the three scoring systems and the threshold effect of each scoring system. Decision curve analysis (DCA) was used to evaluate the application value of each scoring system. RESULTS: The ROC analysis showed that PCIS and pSOFA had a similar predictive value (P=0.182) and that PRISM III and pSOFA had a similar predictive value (P=0.210), while PRISM III had a better predictive value than PCIS (P=0.045). PRISM III had the highest degree of fitting with prognosis, followed by pSOFA and PCIS. The DCA analysis showed that when the risk of death was 0.4 and 0.6 in children with severe sepsis and the three scoring systems were used as the basis for emergency intervention decision-making, pSOFA achieved the highest standardized net benefit, followed by PRISM III and PCIS. CONCLUSIONS: All three scoring systems have a certain value in predicting the prognosis of children with severe sepsis, and pSOFA has a better value than PRISM III and PCIS.

The significance of Notch ligand expression in the peripheral blood of children with hand, foot and mouth disease (HFMD).

BACKGROUND: Hand, foot and mouth disease (HFMD), a virus-induced infectious disease that usually affects infants and children, has an increased incidence in China in recent years. This study attempted to investigate the role of the Notch signaling pathway in the pathogenesis of HFMD. METHODS: Eighty-two children diagnosed with HFMD were enrolled into this study. The HFMD group was further divided into the uncomplicated HFMD and HFMD with encephalitis groups. The control group included 40 children who underwent elective surgery for treatment of inguinal hernias. RESULTS: Children with HFMD displayed significantly reduced CD3+, CD3+CD4+ and CD3+CD8+ cell subsets, but substantially enhanced CD3-CD19+ cell subset (p<0.05 versus control subjects). The expression levels of Notch ligands Dll1 and Dll4 in the peripheral blood of the HFMD group were significantly higher than those in the control group (p<0.05). There were statistically significant differences in CD3+, CD3+CD4+ and CD3-CD19+ cell subsets, but not in Notch ligand expression, between the uncomplicated HFMD and HFMD with encephalitis groups. Dll4 expression in HFMD subjects correlated negatively with the CD3+ and CD3+CD8+ cell subsets (p<0.05), but positively with the CD3-CD19+ cell subset (p<0.05). Furthermore, Dll4 expression in HFMD with encephalitis subjects correlated positively with total white blood cell (WBC) counts and total protein contents in cerebrospinal fluid (CSF) (p<0.05). CONCLUSIONS: The Notch ligand Dll4 exhibits a strong correlation with the CD3+, CD3+CD8+ and CD3-CD19+ cell subsets in children with HFMD, indicating that the Notch signaling may be involved in the development of HFMD by affecting the number and status of peripheral lymphocytes.

[Expression of IGLL1 Gene and Its Clinical Significance in Pediatric T-ALL].

OBJECTIVE: To detect the relative expression of IGLL1 (immunoglobulin lambda-like polypeptide 1) mRNA in bone marrow of children with T-cell acute lymphoblastic leukemia (T-ALL), and analyze its correlation with the clinical characteristics and prognosis of the patients, so as to clarify the clinical significance of IGLL1 in pediatric T-ALL patients. METHODS: A total of 56 pediatric T-ALL patients hospitalized in Children's Hospital of Soochow University from June 2012 to December 2017 and treated with CCLG-ALL 2008 regimen were selected. Transcriptome sequencing technology was used to detect the transcription level of IGLL1 gene in children with T-ALL. According to 25% of the IGLL1 transcription level (cutoff value:448), the enrolled children were divided into IGLL1 low expression group (17 cases) and IGLL1 high expression group (39 cases). Combined with clinical data, the correlation between the expression level of IGLL1 and prognosis of the patients was analyzed. RESULTS: The comparative analysis showed that the transcription level of IGLL1 was not correlated with the clinical characteristics of the patients, such as sex, age, bone marrow blast, white blood cell (WBC) count at initial diagnosis. The 5-year OS rate of patients with high IGLL1 expression was significantly higher than that of patients with low IGLL1 expression (76.9%±6.7% vs 47.1%±12.1%, P =0.018). Further comparison of relapse-free survival (RFS) rate between the two groups showed that the 5-year RFS rate of patients with high IGLL1 expression was higher than that of patients with low IGLL1 expression, but the difference between the two groups was not statistically significant (P =0.095). Multivariate COX analysis was conducted on common clinical prognostic factors (age, sex, WBC count at diagnosis, prednisone response on the 7th day, bone marrow response on the 15th day after treatment) and IGLL1 expression level, and the results showed that IGLL1 expression (P =0.012) and prednisone response (P =0.017) were independent risk factors for overall survival in pediatric T-ALL patients. CONCLUSION: In pediatric T-ALL, the OS rate of children with high expression of IGLL1 gene was significantly higher than that of children with low expression of IGLL1 gene, and the expression level of IGLL1 gene was an independent factor affecting the survival of children with T-ALL, which suggests that IGLL1 is a marker of good clinical prognosis of children with T-ALL.

[Clinical characteristics of 14 critically ill children with 2009 influenza A (H1N1)].

OBJECTIVE: To summarize characteristics and outcomes of critically ill children with 2009 influenza A (H1N1). METHOD: A prospective observational study of 14 critically ill children with 2009 influenza A (H1N1) in pediatric intensive care unit (PICU) in Suzhou between Oct. 1(st) 2009 and Dec. 25(th) 2009. The primary outcome measures included frequency and duration of mechanical ventilation and duration of ICU stay. RESULT: Critical illness occurred in 14 patients with confirmed (n = 14), community-acquired 2009 influenza A virus (H1N1) infection. The mean (SD) age of the 14 patients with confirmed 2009 influenza A (H1N1) was (4.91 ± 4.14) years, 7 were female (50.0%). The median duration from symptom onset to hospital admission was (3.09 ± 1.30) days and from hospitalization to ICU admission was (0.95 ± 0.96) day. All the patients were severely hypoxemic [mean (SD) ratio of PaO2/FiO2 was (191.27 ± 80.58) mm Hg] at ICU admission. ARDS occurred in 11 cases (78.6%). Mechanical ventilation was applied for 10 patients (71.4%). The median duration of ventilation was (12.51 ± 10.03) days and ICU stay was (12.58 ± 10.65) days. The median length of time during which the real-time RT-PCR test results were positive was (17.27 ± 5.57) days; Comorbidities such as iron deficiency anemia, cerebral palsy and congenital heart disease were found in 8 cases (57.1%). The longer length of mechanical ventilation and ICU stay were found in cases with higher admission PRISM III Score and lower Pediatrics Critical Illness Score. CONCLUSION: Critical illness due to 2009 influenza A (H1N1) in Suzhou occurred rapidly after hospital admission and was associated with severe hypoxemia, ARDS, a condition that required prolonged mechanical ventilation. There were myocardial damages in critically ill children with severe 2009 influenza A (H1N1).