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.

Treating GNAO1 mutation-related severe movement disorders with oxcarbazepine: a case report.

Background: GNAO1 variants have been found to be associated with epileptic encephalopathies, developmental delays (DDs), and movement disorders (MDs). Therapies for patients with GNAO1 variants vary. However, treatments for GNAO1-related diseases are still in their infancy. Previous reports suggest that few pharmacological treatments are effective for patients with GNAO1 variant-related MDs. Deep brain stimulation (DBS) treatment appears to be effective, however surgical procedures and equipment failures pose risks to the patients. Effectiveness for oxcarbazepine (OXC) in GNAO1 variant-related MDs is first reported in our study, and it expand the effective drugs for MD treatment. Case Description: We report the case of a 5-year-old male patient with a MD, who suffered from hypotonia and refractory choreoathetosis. The patient was found to have a DD and an intellectual disability. A de-novo variant of the GNAO1 gene (NM_138736: exom6: c.709G>A [p. Glu237Lys]) was identified by whole exome sequencing (WES) when he was 8 months old. The patient visited our hospital at the age of 4 years and 3 months because of fever and recurrent convulsions. Electroencephalogram (EEG) results show abnormal spikes, and magnetic resonance imaging (MRI) showed the enlargement of the lateral ventricles. The administration of tiapride hydrochloride, phenobarbital, midazolam, and hormones had no effect. OXC treatment was then initiated. No MD behaviors, such as rigidity and twisting of the limbs and trunk, or chorea, were observed after 10 days OXC treatment. Eventually, incremental doses of OXC were effective, and our patient achieved good control of his MD. Conclusions: We are the first to demonstrate the role of OXC in alleviating MDs associated with GNAO1 mutations. This report provides a novel possibility for the clinical treatment of this rare disease. To manage MDs associated with GNAO1 mutations, we recommend that OXC treatment be attempted before invasive surgical therapy.

Withdrawal of treatment in a pediatric intensive care unit at a Children's Hospital in China: a 10-year retrospective study.

BACKGROUND: Published data and practice recommendations on end-of-life care generally reflect Western practice frameworks; there are limited data on withdrawal of treatment for children in China. METHODS: Withdrawal of treatment for children in the pediatric intensive care unit (PICU) of a regional children's hospital in eastern China from 2006 to 2017 was studied retrospectively. Withdrawal of treatment was categorized as medical withdrawal or premature withdrawal. The guardian's self-reported reasons for abandoning the child's treatment were recorded from 2011. RESULTS: The incidence of withdrawal of treatment for children in the PICU decreased significantly; for premature withdrawal the 3-year average of 15.1% in 2006-2008 decreased to 1.9% in 2015-2017 (87.4% reduction). The overall incidence of withdrawal of care reduced over the time period, and withdrawal of therapy by guardians was the main contributor to the overall reduction. The median age of children for whom treatment was withdrawn increased from 14.5 months (interquartile range: 4.0-72.0) in 2006 to 40.5 months (interquartile range: 8.0-99.0) in 2017. Among the reasons given by guardians of children whose treatment was withdrawn in 2011-2017, "illness is too severe" ranked first, accounting for 66.3%, followed by "condition has been improved" (20.9%). Only a few guardians ascribed treatment withdrawal to economic reasons. CONCLUSIONS: The frequency of withdrawal of medical therapy has changed over time in this children's hospital PICU, and parental decision-making has been a large part of the change.

Knocking down TRPM2 expression reduces cell injury and NLRP3 inflammasome activation in PC12 cells subjected to oxygen-glucose deprivation.

Transient receptor potential melastatin 2 (TRPM2) is an important ion channel that represents a potential target for treating injury caused by cerebral ischemia. However, it is unclear whether reducing TRPM2 expression can help repair cerebral injury, and if so what the mechanism underlying this process involves. This study investigated the protective effect of reducing TRPM2 expression on pheochromocytoma (PC12) cells injured by oxygen-glucose deprivation (OGD). PC12 cells were transfected with plasmid encoding TRPM2 shRNAS, then subjected to OGD by incubation in glucose-free medium under hypoxic conditions for 8 hours, after which the cells were allowed to reoxygenate for 24 hours. Apoptotic cells, mitochondrial membrane potentials, reactive oxygen species levels, and cellular calcium levels were detected using flow cytometry. The relative expression of C-X-C motif chemokine ligand 2 (CXCL2), NACHT, LRR, and PYD domain-containing protein 3 (NALP3), and caspase-1 were detected using fluorescence-based quantitative reverse transcription-polymerase chain reaction and western blotting. The rates of apoptosis, mitochondrial membrane potentials, reactive oxygen species levels, and cellular calcium levels in the TRPM2-shRNA + OGD group were lower than those observed in the OGD group. Taken together, these results suggest that TRPM2 knockdown reduces OGD-induced neuronal injury, potentially by inhibiting apoptosis and reducing oxidative stress levels, mitochondrial membrane potentials, intracellular calcium concentrations, and NLRP3 inflammasome activation.

Late-onset neonatal sepsis in Suzhou, China.

BACKGROUND: This study aimed to describe the causative organisms of neonatal late-onset sepsis (LOS) and their antimicrobial resistance in Suzhou, Southeast China over a 7-year period. METHODS: We performed a retrospective study on neonates with LOS from Jan1, 2011 to Dec 31, 2017. The demographic, clinical, and laboratory data of neonates with LOS were analyzed. Logistic regression was used to investigate the risk factors with mortality. RESULTS: During the study period, 202 neonates with LOS were finally identified. The most common pathogens were Escherichia coli (29.2%), followed by Klebsiella pneumoniae (19.3%), and Coagulase-negative Staphylococcus (CoNS) (16.8%). Nearly 90% of the K. pneumoniae were resistant to cefazolin and 71.8% to ceftazidime. Thirty-four patients (16.8%) died. Multivariable logistic regression showed that significant predictors of mortality were birth weight < 1500 g, respiratory distress and convulsions. CONCLUSIONS: Gram-negative organisms have an important role in LOS in our region, with high levels of resistance to third-generation cephalosporins. These data may help in the selection of antibiotics for empirical treatment of neonates with sepsis.

A five-year review of Pseudomonas aeruginosa bacteremia in children hospitalized at a single center in southern China.

BACKGROUND: Pediatric Pseudomonas aeruginosa bacteremia is uncommon. It is mostly seen with impaired immune defenses and is most often nosocomially acquired, but it does occasionally occur in the previously healthy. Empiric antibiotics not effective against P. aeruginosa can result in poor outcomes. To determine the risk factors for P. aeruginosa bacteremia, all pediatric cases of P. aeruginosa bacteremia hospitalized at a single center over a 5-year period were reviewed. METHODS: A retrospective cohort study (2006-2010) of P. aeruginosa bacteremia in children under 14 years of age assessing demographics, the presence of underlying diseases, whether nosocomially acquired, clinical and laboratory findings, P. aeruginosa antibiotic susceptibility, antibiotic therapy, and clinical outcomes was performed. RESULTS: Thirty-one children, mean age 46 months, had P. aeruginosa bacteremia (2.6% positive blood cultures); 18 cases were nosocomial, none were multi-resistant, and 13 (42%) had P. aeruginosa isolated from a site other than blood. Ten cases occurred in previously healthy children, all of which were community-acquired, and these children were more likely to present with seizures and gastrointestinal findings than those with underlying conditions. The overall case fatality rate was 52% (16/31); 6/16 were previously healthy. Fatal cases had more leukopenia, elevated aspartate aminotransferase, and lower prealbumin A. Fewer fatal cases (6/16 vs. 14/15) had initial antibiotic coverage effective for P. aeruginosa (p=0.002). No difference in case fatality rate (p>0.05) or antibiotic sensitivity (p>0.05) was found between community-acquired and nosocomial cases. CONCLUSIONS: P. aeruginosa bacteremia in children is rare but often fatal if initial antibiotics do not cover P. aeruginosa. Factors indicative of P. aeruginosa bacteremia remain elusive, especially in previously healthy young children. However, P. aeruginosa bacteremia should be considered if children present with a grave illness, seizures, serious gastrointestinal findings, hypotension, and leukopenia.