Objective To investigate the role and mechanism of RNA-Specific adenosine deaminase 3 (Adar3) in regulating macrophage polarization during Coxsackievirus B3(CVB3)-induced viral myocarditis (VM). Methods Bone marrow-derived macrophages (BMDM) from mice were cultured in vitro and induced into M1/M2 macrophages using interferon-gamma (IFN-γ)/lipopolysaccharide (LPS) or interleukin 4 (IL-4), respectively. The mRNA expression levels of Adar1, Adar2, and Adar3 in each group of cells were assessed by real-time quantitative PCR (qRT-PCR). Specific siRNAs targeting the Adar3 gene were designed, synthesized, and transiently transfected into M2 macrophages. The mRNA levels of M2 polarization-related marker genes-including arginase 1 (Arg1), chitinase 3-like molecule 3 (YM1/Chi3l3), and resistin-like molecule alpha (RELMα/FIZZ1)-were detected by qRT-PCR. RNA sequencing was performed to analyze the signaling pathways affected by Adar3. The expression levels of Wnt/β-catenin signaling pathway were further validated using qRT-PCR and Western blot. The adeno-associated virus overexpressing Adar3 was designed, synthesized, and injected into mice via tail vein. Three weeks later, a myocarditis mouse model was established. After an additional week, the phenotype and function of cardiac macrophages, as well as multiple indicators of VM (including echocardiography, body weight, histopathology and serology) were examined. Additionally, the protein levels of the Wnt/β-catenin signaling pathway were assessed. Results Compared to M0-type macrophages, the expression level of Adar3 was significantly increased in M2-type macrophages. After transfection of Adar3 siRNA, the mRNA levels of Arg1, YM1 and FIZZ1 in M2 macrophages were downregulated. RNA sequencing revealed 149 upregulated genes and 349 downregulated genes. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis and subsequent validation experiments indicated that Adar3 modulated the Wnt/β-catenin signaling pathway. In vivo experiments demonstrated that Adar3 overexpression alleviated the cardiac dysfunction of VM mice. The proportion of M1 macrophages in the heart decreased, while the proportion of M2 macrophages increased. At the same time, the Adar3 overexpression activated the Wnt/β-catenin signaling pathway. Conclusion Adar3 promotes macrophage polarization toward the M2 phenotype by activating the Wnt/β-catenin signaling pathway, thereby alleviating VM.
Animals ; Adenosine Deaminase/metabolism* ; Macrophages/immunology* ; Wnt Signaling Pathway/genetics* ; Myocarditis/immunology* ; Mice ; Coxsackievirus Infections/metabolism* ; Male ; Mice, Inbred BALB C ; Enterovirus B, Human/physiology* ; beta Catenin/genetics*

Objective To investigate the prognostic value of echocardiography indicators combined with se-rum recombinant human arginase 1(ARG1)and glucose-6-phosphate dehydrogenase(G6PD)in children with sepsis.Methods A total of 116 children with sepsis admitted to the hospital from May 2022 to June 2023 were enrolled in the study as the sepsis group.According to the severity of sepsis,the children were further divided into general sepsis group(52 cases),severe sepsis group(38 cases)and septic shock group(26 cases).Ac-cording to the prognosis of the children,the children with sepsis were divided into good prognosis group(84 cases)and poor prognosis group(32 cases).A total of 116 healthy children who underwent physical examina-tion in the hospital during the same period were enrolled as the control group.The left ventricular ejection fraction(LVEF),left ventricular end-diastolic diameter(LVEDD),left ventricular end-diastolic volume(LV-EDV)and early diastolic mitral flow peak velocity(E)were detected by using color Doppler ultrasound.Ser-um ARG1 and G6PD levels were detected by using enzyme-linked immunosorbent assay(ELISA).The echo-cardiographic indexes and serum ARG1 and G6PD levels were compared between the sepsis group and the con-trol group,and among sepsis children with different disease severity and different prognosis.The receiver op-erating characteristic(ROC)curve was used to analyze the predictive value of echocardiographic indexes com-bined with serum ARG1 and G6PD for poor prognosis in children with sepsis.Results Compared with the control group,the sepsis group had significant reductions in LVEF,E,and G6PD(P＜0.05)and significant increases in LVEDD,LVEDV,and ARG1(P＜0.05).With the aggravation of sepsis,the levels of LVEF,E,and G6PD in children with sepsis gradually decreased(P＜0.05),while the levels of LVEDD,LVEDV,and ARG1 gradually increased(P＜0.05).Compared with the good prognosis group,the poor prognosis group had significantly lower levels of LVEF,E,and G6PD(P＜0.05)and significantly higher levels of LVEDD,LV-EDV,and ARG1(P＜0.05).ROC curve analysis showed that the AUC of echocardiographic indexes com-bined with serum ARG1 and G6PD in predicting poor prognosis of children with sepsis was 0.971,and the sensitivity and specificity were 84.4％and 83.2％,respectively.Conclusion The levels of LVEF,E,and G6PD in children with sepsis significantly decreases,and the levels of LVEDD,LVEDV,and ARG1 signifi-cantly increases.Echocardiographic parameters combined with serum ARG1 and G6PD have high predictive value for poor prognosis in children with sepsis.

Objective To analyze the expressions of lymphocytes in the peripheral blood and serum cytokines in the children with secretory otitis media (SOM) and the effects of glucocorticoids interventions.Methods Totally 90 SOM children were selected as case group,and 30 healthy children were selected as control group.The case group was randomly divided into group A (simple oral antibiotic treatment),group B (oral antibiotics combined with local glucocorticoid treatment) and group C (oral antibiotics combined with systemic glucocorticoid treatment),30 cases in each group.The CD4 + T lymphocytes percentage,CD8 + T lymphocytes percentage,the ratio of CD4 +/CD8 + T lymphocytes in peripheral blood and the serum interleukin-2 (IL-2),interferon gamma (IFN-γ),tumor necrosis factor alpha (TNF-α),interleukin-6 (IL-6),interleukin-10 (IL-10) levels were detected and compared between case group and control group.The air conduction auditory thresholds under different frequency of the patients in group A,group B and group C were examined and compared.Results The CD4 + T lymphocytes percentage,the ratio of CD4 +/CD8 + T lymphocytes in peripheral blood and the serum IL-2,IFN-γ,TNF-α,IL-6,IL-10 levels of the patients in the case group were significantly higher than those in the control group (P ＜ 0.0 5).After the treatment,the above indicators of the children in group A,group B and group C decreased,and there were significant differences before and after the treatment between two groups (P ＜ 0.05).The air conduction auditory thresholds under different frequency of the patients in group B or group C were significantly lower than those in group A (P ＜ 0.05),while there were no significant differences in the air conduction auditory thresholds under different frequency between the children in group B and group C (P ＞ 0.05).Conclusion The patients with SOM show imbalanced cell immune function and cytokines expressions in the peripheral blood.The combination of glucocorticoids therapy and routine antibiotic therapy can effectively improve the immune function and the therapeutic effects.

Objective To analyze the expressions of lymphocytes in the peripheral blood and serum cytokines in the children with secretory otitis media (SOM) and the effects of glucocorticoids interventions.Methods Totally 90 SOM children were selected as case group,and 30 healthy children were selected as control group.The case group was randomly divided into group A (simple oral antibiotic treatment),group B (oral antibiotics combined with local glucocorticoid treatment) and group C (oral antibiotics combined with systemic glucocorticoid treatment),30 cases in each group.The CD4 + T lymphocytes percentage,CD8 + T lymphocytes percentage,the ratio of CD4 +/CD8 + T lymphocytes in peripheral blood and the serum interleukin-2 (IL-2),interferon gamma (IFN-γ),tumor necrosis factor alpha (TNF-α),interleukin-6 (IL-6),interleukin-10 (IL-10) levels were detected and compared between case group and control group.The air conduction auditory thresholds under different frequency of the patients in group A,group B and group C were examined and compared.Results The CD4 + T lymphocytes percentage,the ratio of CD4 +/CD8 + T lymphocytes in peripheral blood and the serum IL-2,IFN-γ,TNF-α,IL-6,IL-10 levels of the patients in the case group were significantly higher than those in the control group (P ＜ 0.0 5).After the treatment,the above indicators of the children in group A,group B and group C decreased,and there were significant differences before and after the treatment between two groups (P ＜ 0.05).The air conduction auditory thresholds under different frequency of the patients in group B or group C were significantly lower than those in group A (P ＜ 0.05),while there were no significant differences in the air conduction auditory thresholds under different frequency between the children in group B and group C (P ＞ 0.05).Conclusion The patients with SOM show imbalanced cell immune function and cytokines expressions in the peripheral blood.The combination of glucocorticoids therapy and routine antibiotic therapy can effectively improve the immune function and the therapeutic effects.