Background: Mice are useful small animal models to study the pathogenesis of SARS-CoV-2 infection. As the ancestral SARS-CoV-2 strains did not utilize murine Ace2 as a receptor, wild-type mice were not susceptible to the SARS-CoV-2 infection. Infection of human ACE2-expressing transgenic mice with SARS-CoV-2 induces fatal encephalitis, which is not commonly observed in humans. We and others have previously demonstrated the ability of the SARS-CoV-2 Beta variant to productively infect wild-type mice. Herein, we employed RNA-seq to investigate the transcriptomic landscapes in the lungs after the infection of wild-type mice with SARS-CoV-2 Beta variant. Methods: We intranasally infected 6-week-old wild-type C57BL/6J mice with the SARS-CoV-2 (B.1.351 strain) and collected lungs at 3- and 6-days post-infection for RNA-sequencing. We used the Limma-Voom package to identify differentially expressed genes (DEGs) and the fgsea package for pathway enrichment analysis. We used Cytoscape to identify hub genes and gene networks. Lastly, we employed RT-qPCR and multiplex assay to validate the RNA-seq data. Results: Using a cutoff of an adjusted p-value below 0.05 and an absolute log2 fold change value greater than 0.75, we identified 285 DEGs on day 3 and 46 DEGs on day 6. The canonical pathways analysis showed that several key pathways such as apoptosis and cytokine response were upregulated in the infected lungs. Protein-protein interaction analyses identified innovative target genes such as if11, Ccna2, and Aurkb. We also identified the top 10 hub genes that included Prc1, Ube2c, Ccnb2, Ncapg, Aurkb, Cep55, Mki67, Dlgap5, Ccna2, and Kif11. RT-qPCR analysis for Tnfa, Il6, Ccl2, and Ccl3 further validated the RNA-seq analysis. Consistent with gene expression results, we detected significantly increased protein levels of various inflammatory mediators such as IL-6, CCL2, CXCL2, and CXCL10 in the infected lungs. Conclusions This is the first transcriptomic analysis of the lungs of wild-type mice infected with a clinical isolate of SARS-CoV-2. Our findings provide a further understanding of the pathogenic events that occur in this mouse model of SARS-CoV-2 infection.

Purpose: We evaluated the effectiveness of early start of long-acting insulin during management of diabetic ketoacidosis (DKA) in pediatric patients. Methods: Patients with DKA were randomly assigned to receive either a traditional DKA management protocol or concurrent administration of subcutaneous (SC) long-acting insulin alongside intravenous insulin during DKA treatment. The primary outcomes were duration of insulin infusion and adverse effects of the intervention, mainly hypoglycemia and hypokalemia. Results: For this study, 100 pediatric patients with DKA were enrolled, 50 in each group (group I received the conventional DKA management and group II received conventional DKA management plus SC long-acting insulin once daily). Patients in group II showed a significant reduction in both duration and dose of insulin infusion compared to group I, with a median (interquartile range) of 68.5 hours (45.00–88.25 hours) versus 72 hours (70.25–95.5 hours) (P=0.0001) and an insulin dose of 3.48±1.00 units/kg versus 4.04±1.17 units/kg (P=0.016), respectively. Concurrent administration of SC long-acting insulin with intravenous insulin during DKA treatment was associated with a decreased risk of hypoglycemia (number of hypoglycemia events: group I, 22 events; group II, 12 events, P=0.029), with no increased risk of hypokalemia compared to the control group (number of hypokalemia events: group I, 12 events; group II, 19 events, P=0.147). Conclusion The current study showed that coadministration of SC long-acting insulin in addition to the usual insulin infusion during DKA management in the pediatric population can lead to a shorter duration of insulin infusion. In addition, this approach is not associated with increased risk of hypoglycemia or hypokalemia. Moreover, coadministration of long-acting insulin may be associated with a decreased incidence of hypoglycemia.

Purpose: We evaluated the effectiveness of early start of long-acting insulin during management of diabetic ketoacidosis (DKA) in pediatric patients. Methods: Patients with DKA were randomly assigned to receive either a traditional DKA management protocol or concurrent administration of subcutaneous (SC) long-acting insulin alongside intravenous insulin during DKA treatment. The primary outcomes were duration of insulin infusion and adverse effects of the intervention, mainly hypoglycemia and hypokalemia. Results: For this study, 100 pediatric patients with DKA were enrolled, 50 in each group (group I received the conventional DKA management and group II received conventional DKA management plus SC long-acting insulin once daily). Patients in group II showed a significant reduction in both duration and dose of insulin infusion compared to group I, with a median (interquartile range) of 68.5 hours (45.00–88.25 hours) versus 72 hours (70.25–95.5 hours) (P=0.0001) and an insulin dose of 3.48±1.00 units/kg versus 4.04±1.17 units/kg (P=0.016), respectively. Concurrent administration of SC long-acting insulin with intravenous insulin during DKA treatment was associated with a decreased risk of hypoglycemia (number of hypoglycemia events: group I, 22 events; group II, 12 events, P=0.029), with no increased risk of hypokalemia compared to the control group (number of hypokalemia events: group I, 12 events; group II, 19 events, P=0.147). Conclusion The current study showed that coadministration of SC long-acting insulin in addition to the usual insulin infusion during DKA management in the pediatric population can lead to a shorter duration of insulin infusion. In addition, this approach is not associated with increased risk of hypoglycemia or hypokalemia. Moreover, coadministration of long-acting insulin may be associated with a decreased incidence of hypoglycemia.

Purpose: We evaluated the effectiveness of early start of long-acting insulin during management of diabetic ketoacidosis (DKA) in pediatric patients. Methods: Patients with DKA were randomly assigned to receive either a traditional DKA management protocol or concurrent administration of subcutaneous (SC) long-acting insulin alongside intravenous insulin during DKA treatment. The primary outcomes were duration of insulin infusion and adverse effects of the intervention, mainly hypoglycemia and hypokalemia. Results: For this study, 100 pediatric patients with DKA were enrolled, 50 in each group (group I received the conventional DKA management and group II received conventional DKA management plus SC long-acting insulin once daily). Patients in group II showed a significant reduction in both duration and dose of insulin infusion compared to group I, with a median (interquartile range) of 68.5 hours (45.00–88.25 hours) versus 72 hours (70.25–95.5 hours) (P=0.0001) and an insulin dose of 3.48±1.00 units/kg versus 4.04±1.17 units/kg (P=0.016), respectively. Concurrent administration of SC long-acting insulin with intravenous insulin during DKA treatment was associated with a decreased risk of hypoglycemia (number of hypoglycemia events: group I, 22 events; group II, 12 events, P=0.029), with no increased risk of hypokalemia compared to the control group (number of hypokalemia events: group I, 12 events; group II, 19 events, P=0.147). Conclusion The current study showed that coadministration of SC long-acting insulin in addition to the usual insulin infusion during DKA management in the pediatric population can lead to a shorter duration of insulin infusion. In addition, this approach is not associated with increased risk of hypoglycemia or hypokalemia. Moreover, coadministration of long-acting insulin may be associated with a decreased incidence of hypoglycemia.