The aim of this study was to investigate the expression profiles of microRNAs (miRNAs) in pediatric asthma and to determine candidate miRNAs responsible for the pathogenesis of this disease. Microarrays were used to detect the differences in the miRNA expression levels between asthmatic children and controls. Airway inflammation was evaluated by cell counting and tissue biopsy in an ovalbumin (OVA)-induced murine asthma model. Real-time polymerase chain reaction (PCR) was used to verify the differentially expressed miRNAs. The targets of the identified miRNAs were analyzed by bioinformatic analysis. The sprouty-related protein with an EVH1 domain-2 (Spred-2) protein content was assessed by western blotting. Differences were observed in the expression of miRNAs between the asthmatic children and controls. Upregulation of miRNA-221 and miRNA-485-3p in pediatric asthmatics and murine asthma models were verified by real-time PCR. Spred-2, a predicted target of miRNA-221 and miRNA-485-3p, was downregulated in murine asthma models. Upregulation of miRNA-221 and miRNA-485-3p may regulate the pathogenesis of asthma.

The imaging findings of Mycoplasma pneumoniae pneumonia (MPP) vary; however, few studies have focused on the relationship of imaging classification with clinical manifestations and outcomes. To prospectively investigate whether chest imaging classification in Mycoplasma pneumoniae pneumonia (MPP) is associated with its clinical features and outcomes. A total of 1,401 hospitalized children with MPP were enrolled from January 2019 to December 2021. Imaging findings were categorized as bronchopneumonia and consolidation/atelectasis according to X-ray, and bronchopneumonia, consolidation/atelectasis, bronchiolitis, and mosaic pattern according to computed tomography (CT). Clinical characteristics and outcomes of patients with different imaging classifications were prospectively analyzed based on electronic medical records. Bronchopneumonia was the most common finding (59.6%), while consolidation/atelectasis was the most severe group. Clinical manifestations and laboratory indicators for the consolidation/atelectasis group included serious abnormalities. Further, outcomes of the patients were worse, including having longer total durations of fever and hospitalization, greater hospitalization expenses, and a higher likelihood of developing refractory MPP, necrotizing pneumonia, and bronchiolitis obliterans (BO) in this group. The incidence of bronchiolitis, a disease characterized by a high prevalence of fever, moist rales, and an atopic constitution, tended to increase after the coronavirus disease pandemic and predisposed patients to BO. A mosaic pattern occurred in allergic and young individuals, with wheezing as the main manifestation, with patients having relatively mild symptoms and good outcomes. Different imaging classifications have different clinical features and clinical outcomes; thus, formulating an imaging-based classification system is of great clinical value.

Background: The association between sensitizations and severity of allergic diseases is controversial. Objective: This study was to investigate the association between severity of asthma and rhinitis and degree of specific allergic sensitization in allergic patients in China. Method: A cross-sectional survey was performed in 6304 patients with asthma and/or rhinitis from 4 regions of China. Patients completed a standardized questionnaire documenting their respiratory and allergic symptoms, their impact on sleep, daily activities, school and work. They also underwent skin prick tests with 13 common aeroallergens. Among the recruited subjects, 2268 provided blood samples for serum measurement of specific IgE (sIgE) against 16 common aeroallergens. Results: Significantly higher percentage of patients with moderate-severe intermittent rhinitis were sensitized to outdoor allergens while percentage of patients sensitized to indoor allergens was increased with increasing severity of asthma. Moderate-severe intermittent rhinitis was associated with the skin wheal size and the level of sIgE to Artemisia vulgaris and Ambrosia artemisifolia (p < 0.001). Moderate-severe asthma was associated with increasing wheal size and sIgE response to Dermatophagoides (D.) pteronyssinus and D. farinae (p < 0.001). Moderate-severe rhinitis and asthma were also associated with increase in number of positive skin prick test and sIgE. Conclusions: Artemisia vulgaris and Ambrosia artemisifolia sensitizations are associated with the severity of intermittent rhinitis and D. pteronyssinus and D. farinae sensitizations are associated with increasing severity of asthma in China. Increase in number of allergens the patients are sensitized to may also increase the severity of rhinitis and asthma.

Neutrophil extracellular traps (NETs) are web‑like structures made of chromatin and have been identified to have a role in the host's immune defense. Differentiated human promyelocytic leukemia HL‑60 cells (dHL‑60) have been used to study the mechanisms of NETs formation, as neutrophils have a short lifespan that limits their use. However, dHL‑60 cells are inefficient at generating NETs and therefore are not ideal replacements for neutrophils in studying of NET formation. In the present study, the optimal cell culture conditions and differentiation time that result in the most effective release of NETs from dHL‑60 cells upon stimulation were determined. HL‑60 cells were cultured in serum (FBS) or serum‑free (X‑VIVO) medium and differentiated using all‑trans retinoic acid (ATRA) or dimethyl sulfoxide (DMSO). dHL‑60 cells were stimulated with phorbol 12‑myristate 13‑acetate (PMA) or Ca2+ ionophore (CI). Cell differentiation and apoptosis, as well as the formation of reactive oxygen species (ROS) and citrullinated histone H3 (citH3) were analyzed using flow cytometry. NETs were visualized using fluorescence microscopy and NET quantification was performed using PicoGreen. Induction of HL‑60 cells for five days produced the best results in terms of differentiation markers and cell viability. Both ATRA‑ and DMSO‑induced dHL‑60 cells were able to release NETs upon PMA and CI stimulation; dHL‑60 cells in serum‑free medium produced more NETs than those in serum‑containing medium. DMSO‑dHL‑60 (X‑VIVO) cells were most efficient at producing NETs and ROS upon stimulation with PMA, while ATRA‑dHL‑60 (X‑VIVO) cells were most efficient at producing NETs and citH3 upon stimulation with CI. It was concluded that DMSO‑dHL‑60 (X‑VIVO) may be a model for the study of ROS‑high NETosis and ATRA‑dHL‑60 (X‑VIVO) may be suitable for ROS‑low NETosis.

Now there is no clinical scale for early prediction of refractory Mycoplasma pneumoniae pneumonia (RMPP). The aim of this study is to identify indicators and develop an early predictive scale for RMPP in hospitalized children. First we conducted a retrospective cohort study of children with M. pneumoniae pneumonia admitted to Children’s Hospital of Nanjing Medical University, China in 2016. Children were divided into two groups, according to whether their pneumonia were refractory and the results were used to develop an early predictive scale. Second we conducted a prospective study to validate the predictive scale for RMPP in children in 2018. 618 children were included in the retrospective study, of which 73 with RMPP. Six prognostic indicators were identified and included in the prognostic assessment scale. The sensitivity of the prognostic assessment scale was 74.0% (54/73), and the specificity was 88.3% (481/545) in the retrospective study. 944 children were included in the prospective cohort, including 92 with RMPP, the sensitivity of the prognostic assessment scale was 78.3% (72/92) and the specificity was 86.2% (734/852). The prognostic assessment scale for RMPP has high diagnostic accuracy and is suitable for use in standard clinical practice.