To construct a nomogram model for analyzing risk factors of wheezing in infants with Mycoplasma pneumoniae pneumonia (MPP), so as to provide evidence for clinical intervention. A retrospective analysis was performed on clinical data of 569 infants with MPP hospitalized in the Department of Respiratory Medicine and Department of Infectious Diseases, Children’s Hospital of Soochow University. They were divided into a wheezing group (204 cases) and a non-wheezing group (365 cases) according to symptoms. Demographic characteristics, clinical manifestations, laboratory examinations, bronchoscopy frequency and chest imaging findings were collected. Differences in clinical and laboratory indicators between the two groups were compared. Least Absolute Shrinkage and Selection Operator (LASSO) regression was used for preliminary screening, followed by Logistic regression to identify independent risk factors for wheezing in infants with MPP. Finally, a nomogram prediction model was established, and its performance was evaluated by receiver operating characteristic (ROC) curve and decision curve analysis (DCA). 1. Univariate and multivariate analysis of predictors for wheezing in MPP: Univariate Logistic regression screening revealed 16 potential risk factors: fever, cough, history of wheezing, history of eczema, WBC, N#, L#, PLT, CRP, IgG, IgM, CKMB, D-D (ug/L), HRV, HRSV, HMPV (P < 0.05). Multivariate Logistic regression showed that history of wheezing (OR = 5.894, 95

Objective To analyse the characteristic features of infants with pneumonia who test positive for serum milk-specific immunoglobulin E (sIgE) and to provide a reference for the diagnosis, management and prevention of the condition. Methods We retrospectively analysed data from 284 infants admitted to our hospital with pneumonia between January 2017 and December 2020 who underwent serum allergen testing. Based on the results, patients were categorised into three groups: pure milk sIgE-positive; mixed milk sIgE-positive; and allergen sIgE-negative. We then compared the general conditions, clinical characteristics, laboratory tests, imaging results and pathogenic data across these groups. Results Among the patient population, 16.20% (46/284) tested positive for pure milk sIgE, 32.75% (93/284) tested positive for mixed milk sIgE and 51.06% (145/284) were negative for any allergen sIgE. Statistically significant differences were observed among the three groups in terms of general conditions, breastfeeding status, pre-existing respiratory infections and history of respiratory infections (>3 times) ( p < 0.05 for each). The median length of hospital stay was longer in the pure milk sIgE-positive group and the mixed milk sIgE-positive group (8 [range 7–10] days) compared with the allergen sIgE-negative group (8 [range 6–9] days) ( p < 0.05). The eosinophil counts of the mixed milk sIgE-positive group were significantly higher than in the other two groups ( p < 0.05). Haemophilus influenzae of the pure milk sIgE-positive group was significantly higher than in the other two groups ( p < 0.05). Conclusion The presence of mixed milk sIgE allergens does not exacerbate clinical symptoms. However, infants who test positive solely for milk sIgE and have pneumonia require longer hospitalisation. This suggests that extra attention is necessary for infants with milk allergies when they develop pneumonia.

Cow's milk protein allergy is an adverse immune reaction to proteins found in cow's milk, primarily casein and whey, affecting artificially fed, breastfed and mixed-fed infants. The immunological mechanisms involved lead to diverse clinical presentations, most commonly affecting the digestive, respiratory, and integumentary systems. Diagnosis relies primarily on clinical evaluation due to the absence of specific diagnostic tests, making accurate identification crucial to prevent misdiagnosis or underdiagnosis. Treatment requires strict avoidance of cow's milk proteins in the diets of both children and breastfeeding mothers, with close monitoring of nutritional status during long-term management. Recent advancements in treatment, including the use of probiotics, provide new options for improving clinical outcomes. This narrative review aims to provide clinicians with evidence to standardise diagnosis and treatment, improve food allergy management by non-allergy specialists and develop accurate feeding recommendations.

Although microwave ablation (MWA) is an important curative therapy in colorectal cancer liver metastasis, recurrence still occurs clinically. Our previous studies have shown that the expression of programmed cell death 1 ligand 1 (PD-L1) is upregulated following MWA, suggesting that MWA combined with anti-PD-L1 treatment can serve as a promising clinical therapeutic strategy against cancer. Using MWA-treated preclinical mice models, MWA combined with αPD-L1 treatment decreased tumor growth and prolonged overall survival (OS). Furthermore, through flow cytometry and single-cell RNA sequencing analysis, we determined that the MWA plus αPD-L1 therapy significantly suppressed CD8+ T cell exhaustion and enhanced their effector function. A significant increase in γ-interferon (IFN-γ) stimulated transcription factors, specifically Irf8, was observed. This enhancement facilitated the polarization of tumor-associated macrophages (TAM1s and TAM2s) through the nuclear factor-κB/JAK-STAT1 signaling pathway. Furthermore, the combination therapy stimulated the production of CXC motif chemokine ligand (CXCL9) by TAM1s and tumor cells, potentially increasing the chemotaxis of CD8 T cells and Th1 cells. Knocking out Cxcl9 in MC38 tumor cells or using CXCL9 blockade enhanced tumor growth of untreated tumors and shortened OS. Taken together, our study showed that blocking the IFN-γ-Cxcl9-CD8+ T axis promoted tumor progression and discovered a potential involvement of IRF8-regulated TAMs in preventing T cell exhaustion. Collectively, we identified that the combination of MWA with anti-PD-L1 treatment holds promise as a therapeutic strategy to rejuvenate the immune response against tumors. This merits further exploration in clinical studies.

Background: Neutrophilic asthma is characterized by the predominant infiltration of neutrophils in airway inflammation. Objective: To explore the therapeutic potential of an antibody against the inducible T cell co-stimulator ligand (ICOSL) in a mouse model of neutrophilic asthma. Methods: Female BALB/c mice were randomly assigned to different groups. They were then injected with ovalbumin (OVA)/lipopolysaccharides (LPS) to induce neutrophilic asthma. The mice were then treated with either anti-ICOSL (the I group), control IgG (the G group), or no treatment (the N group). Additionally, a control group of mice received vehicle PBS and was labeled as the C group (n=6 per group). One day after the last allergen exposure, cytokine levels were measured in plasma and bronchoalveolar lavage fluid (BALF) using ELISA. After analyzing and categorizing BALF cells, the lung tissues were examined histologically and immunohistochemically. Results: Administering anti-ICOSL resulted in a significant decrease in the total number of inflammatory infiltrates and neutrophils found in BALF. Moreover, it led to a decrease in the levels of interleukin (IL)-6, IL-13, and IL-17 in both BALF and plasma. Additionally, there was an increase in IFN-gamma levels in the BALF of asthmatic mice (p<0.05 for all). Treatment with anti-ICOSL also reduced lung interstitial inflammation, mucus secretion, and ICOSL expression in asthmatic mice. Conclusion: The treatment of anti-ICOSL effectively improved lung interstitial inflammation and mucus secretion in mice with neutrophilic asthma by restoring the balance of Th1/Th2/Th17 responses. These findings indicate that blocking the ICOS/ICOSL signaling could be an effective way to manage neutrophilic asthma.

Background Refractory mycoplasma pneumoniae pneumonia (RMPP) is a serious mycoplasma pneumoniae infection and is difficult to diagnose early. The levels of serum soluble B7-dendritic cell (sB7-DC) in children with mycoplasma pneumoniae pneumonia (MPP) were assessed to explore the clinical significance of sB7-DC levels in RMPP. Methods A total of 65 patients with mycoplasma pneumoniae pneumonia (MPP) were enrolled in this study between January 2017 and December 2018. The patients were divided into the general mycoplasma pneumoniae pneumonia (GMPP) (n=30) and RMPP groups (n=35); the data of 20 normal children served as a control group (n=20). An enzyme-linked immunoassay kit was used to detect the expression of soluble B7-dendritic cell (sB7-DC) and other inflammatory factors. Binary logistic regression was performed to identify the independent predictors of RMPP. Receiver operating characteristic (ROC) curves were drawn to evaluate the value of each independent risk factor in the early diagnosis of RMPP. Results The results showed that compared to the GMPP group, children in the RMPP group had a significantly longer hospital stay and had a significantly longer fever duration (P<0.05). The values of interferon-gamma (IFN-γ), interleukin 17 (IL-17), and sB7-DC in the RMPP group were significantly higher than those in the normal control and GMPP groups (all P<0.05). The results of the correlation analysis showed that sB7-DC was positively correlated with IFN-γ and IL-17 and these indicators could be used in combination to evaluate the severity of the disease. The binary logistic regression analysis identified IL-17 and sB7-DC as independent risk factors for RMPP (P<0.05). The ROC curve analysis showed that the cut-off values of IL-17 and sB7-DC were 309.6 pg/L and 1,109.7 pg/mL, respectively. The areas under the curve (AUCs) of IL-17 and sB7-DC were 0.741 and 0.794, respectively. The sensitivity of IL-17 to RMPP prediction was 83.3%, and the specificity was 62.9%. The sensitivity and specificity of sB7-DC to RMPP were 86.7% and 62.9%, indicating that sB7-DC had the highest predictive power for RMPP. Conclusions The level of serum sB7-DC may play an important role in the early diagnosis of RMPP. Our research results provide a theoretical basis for the early diagnosis of RMPP.

This study aimed to explore the effects of forkhead box P2 gene (Foxp2) on T-helper 9 (Th9) differentiation in asthmatic mice. An in vivo asthmatic mouse model was induced with ovalbumin (OVA). An in vitro model was established by culturing CD4+ T cells with TGF-β, IL-4, and anti-IFN-γ. ELISA, flow cytometry, qRT-PCR and Western blot were performed to examine IL-9 secretion, Th9 cell number, and Th9 cell transcription factor expression, respectively. Pathological changes in lung tissues and airway mucus secretion were assessed with HE and PAS glycogen staining. Anti-IL-9 mAb reversed the elevation in Th9 cells and IL-9 expression in lung tissues and bronchoalveolar lavage fluid (BALF) of asthmatic mice. Foxp2 was downregulated in BALF and lung tissue of asthmatic mice and Th9 cells. Overexpression of Foxp2 inhibited Th9 cell differentiation in vitro and improved airway inflammation in vivo. Our study suggests that overexpression of Foxp2 attenuates allergic asthma by inhibiting Th9 cell differentiation.

Purpose To investigate the impact of COVID-19 on the treatment of children with congenital diaphragmatic hernia (CDH). Methods We retrospectively collected and compared the data of patients with CDH admitted between January 1, 2020 and December 31, 2021(study group) with the CDH patients admitted before the pandemic between January 1, 2018 and December 31, 2019 (control group). Results During the pandemic, 41 patients with CDH diagnosed prenatally were transferred to our hospital, and 40 underwent surgical repair. The number of patients treated in our hospital increased by 24.2% compared with the 33 patients before the pandemic. During the pandemic, the overall survival rate, postoperative survival rate and recurrence rate were 85.4%, 87.5% and 7.3%, respectively, and there were no significant differences compared with the control group (75.8%, 83.3% and 9.1%, respectively). The average length of hospital stay in patients admitted during the pandemic was longer than that in the control group (31 days vs. 16 days, P < 0.001), and the incidence of nosocomial infection was higher than that in the control group (19.5% vs. 3%, P = 0.037). Conclusions CDH patients confirmed to be SARS-CoV-2 infection-free can receive routine treatment. Our data indicate that the implementation of protective measures during the COVID-19 pandemic, along with appropriate screening and case evaluation, do not have a negative impact on the prognosis of children.

The role of micro RNAs (miRNAs) in asthma remains unclear. In this study, we examined the role of miRNA in targeting FOXO1 in asthma. Results showed that miR-493-5p was one of the differentially expressed miRNAs in the PBMCs of asthmatic children, and was also associated with Th cell differentiation. The miR-493-5p expression decreased significantly in the OVA-induced asthma mice than the control groups. The miR-493-5p mimic inhibited the expression of the IL-9, IRF4 and FOXO1, while the inhibitor restored these effects. Moreover, the Dual-Luciferase analysis results showed FOXO1 as a novel valid target of miR-493-5p. According to the rescue experiment, miR-493-5p inhibited Th9 cell differentiation by targeting FOXO1. Then the exosomes in association with the pathogenesis of asthma was identified. Various inflammatory cells implicated in asthmatic processes including B and T lymphocytes, DCs, mast cells, and epithelial cells can release exosomes. Our results demonstrated that the DC-derived exosomes can inhibit Th9 cell differentiation through miR-493-5p, thus DC-derived exosomal miR-493-5p/FOXO1/Th9 may serve as a potential therapeutic target in the development of asthma.

Abstract Background MicroRNAs play an important role in T cell responses. However, how microRNAs regulate Th cells in asthma remains poorly defined. Objective In this study, we investigated the mechanism and pathways of miR‐29b regulating Th cells in asthma, in order to find new targets for asthma. Methods We detected miR‐29b, B7‐H3 and STAT3 in the peripheral blood of children with asthma, explored the relationship between these molecules and their effects on T cells through in vitro cell culture, and verified it by animal model. Results MiR‐29b levels were decreased in the peripheral blood mononuclear cells from children with asthma. Vitro studies found that the expression of miR‐29b in macrophages was decreased and the expression of B7‐H3 and STAT3 was increased after house dust mite (HDM) stimulation. After down‐regulation of miR‐29b in macrophages, the expressions of B7‐H3 and STAT3 in macrophages were increased and T cells differentiate into Th2 cells. After the addition of B7‐H3 or STAT3 antibodies, the differentiation of naive T cells into Th2 cells was reduced. In OVA induced mice asthmatic model, after the up‐regulation of miR‐29b in lung, the expression of B7‐H3 and STAT3 decreased in the lung tissues of mice, and the expression of Th2 cells and type II cytokine decreased simultaneously. The pathological changes of lung tissues were also alleviated. Conclusion The expression of miR‐29b is decreased in asthmatic children. MiR‐29b can inhibit Th2 cell differentiation by inhibiting B7‐H3 and STAT3 pathways at the same time, and reduce asthmatic immune inflammation.

Background Programmed death-ligand 1 (PD-L1) is an important immune checkpoint inhibitor. Recent studies suggest that the PD-L1-mediated pathway may be a promising target in allergic asthma. However, the mechanism by which PD-L1 represses neutrophilic asthma (NA) remains unclear. In this study, we examined correlations between the expression of PD-L1 and the production of T helper cell type 1 (Th1), T helper cell type 2 (Th2), and T helper cell type 17 (Th17) cells in pediatric patients with NA and a mouse model. Methods The clinical samples of 26 children with asthma and 15 children with a bronchial foreign body were collected over a period of 12 months by the Children’s Hospital of Soochow University. An experimental mouse model of asthma was established to study NA. An enzyme-linked immunoassay (ELISA) was used to assess soluble PD-L1 (sPD-L1) and cytokines [e.g., interleukin (IL)-4, IL-6, interferon gamma (IFN-γ), IL-17 and granulocyte-macrophage colony-stimulating factor (GM-CSF)] in bronchoalveolar lavage fluid (BALF). Results NA patients had significantly higher levels of sPD-L1, IL-6, IL-17, and GM-CSF in their BALF than non-NA and control patients (P<0.05). In a murine model of asthma, the positive rate and fluorescence intensity of PD-L1 in the NA group and the immunoglobulin G (IgG)-treated NA group were higher than in the PD-L1 antibody (Ab)-treated NA group and the phosphate-buffered saline (PBS) control group (P<0.05). In the plasma and the BALF of the NA group and the IgG-treatment NA group, the levels of IL-17, IL-4, tumor necrosis factor alpha (TNF-α), and granulocyte colony-stimulating were higher than those in the PBS control group (P<0.05). The histopathological examination of lung tissues from all mice groups showed that a large number of inflammatory cells were found around the airway in the NA group and the IgG-treatment group. Conclusions PD-L1 may contribute to the Th17/IL-17 immune response, which is associated with neutrophilic inflammation and asthma. A PD-L1 blockade reduces pulmonary neutrophils and mucus production.

Background: To explore the roles of Toll-like receptor (TLR)2 in Th2 cytokine production and immunoglobulin (Ig) class switching following ovalbumin (OVA) sensitization.Methods: TLR2 -/- and wild-type C57BL/6 mice were sensitized by intraperitoneal injection with OVA. Lung pathology was assessed by hematoxylin and eosin staining. Abundance of interleukin (IL)4, IL5, IL13, and IL21 transcripts in the lungs was quantified by RT-PCR. OVA-specific IgG1, IgG2a, IgG2b, IgE and IgM were quantified by enzyme-linked immunosorbent assay. Phosphorylated signal transducer and activator of transcription (STAT)3 in lung tissue was detected by immunohistochemistry staining and nuclear factor (NF) κB activation was measured by immunofluorescence staining. STAT3 activation was inhibited using cryptotanshinone (CPT) treatment. Germline transcripts (Iμ-Cμ, Iγ-Cγ, Iα-Cα or Iε-Cε), post-recombination transcripts (Iμ-Cγ, Iμ-Cα or Iμ- Cε) and mature transcripts (V H DJ H -Cγ, V H DJ H -Cα or V H DJ H -Cε) were analyzed from splenic B cells of OVA-sensitized wild-type mice (with or without CPT treatment) and TLR2 -/- mice (with or without IL21 treatment). Results: The lungs of TLR2 -/- mice showed a lesser degree of inflammation than wild-type mice after OVA sensitization. Following OVA sensitization, levels of IL4, IL13, and IL21, but not IL5, were significantly lower in TLR2 -/- compared with wild-type mice. Moreover, OVA-specific IgG1 and IgE titers were markedly lower and higher, respectively, in TLR2 -/- mice. TLR2 deficiency inhibited STAT3 activation but not NF-κB p65 activation. CPT treatment reduced IgG1 titers via inhibition of Stat3 phosphorylation. Both TLR2 knockout and CPT treatment reduced the frequencies of Iγ1-Cγ1, Iγ3-Cγ3 and Iα-Cα transcripts, but IL21 treatment compensated for the effects of TLR2 deficiency.Conclusion: These results suggest a role of TLR2 in restricting OVA-sensitized lung inflammation via promotion of IgG1 and inhibition of IgE class switching regulated by IL21 and STAT3.

Background Bronchiolitis is a clinical syndrome commonly encountered in practice, particularly among infants and young children. To investigate the prevalence of pathogens in hospitalized infants with bronchiolitis and study the relationship between the clinical characteristics and pathogens. Methods We investigated the respiratory specimens and clinical data of 1012 children with bronchiolitis who were treated at the Children’s Hospital of Soochow University between November 2011 and December 2018. The nasopharyngeal aspirates were examined by direct immunofluorescence assay or polymerase chain reaction (PCR) to detect viruses and by PCR and enzyme-linked immunosorbent assay to detect Mycoplasma pneumoniae (MP). Results Of the 1012 children with bronchiolitis, 842 (83.2%) were detected at least a pathogen. 614 (60.7%) had single viral infections, 91 (9.0%) had MP infections, 70 (6.9%) had multiple viral infections, and 67 (6.6%) had mixed viral and MP infection. The most common pathogens detected were respiratory syncytial virus (RSV) (44.4%), MP (15.6%), and human rhinovirus (HRV) (14.4%). RSV was the most common pathogen detected in children less than 6 months. Coinfection was detected in 13.5% (137/1012) of the children, but it was less common in children less than 6 months. The age of children with single virus infection was the youngest. Children with single virus infection had a higher proportion of oxygen therapy compared with single MP infection. Conclusions The most common pathogen detected in children with bronchiolitis is RSV, followed by MP and HRV. Co-infections lead to prolonged illness and worsening of the symptoms Keywords: Etiology, Bronchiolitis, Infant, Respiratory syncytial virus, Mycoplasma pneumoniae

OBJECTIVE:Pneumonia is an infectious pulmonary disease with a high morbidity and mortality. It has been reported that multiple long noncoding RNAs (LncRNAs) are involved in the progression of pneumonia, such as LncRNA SNHG16. However, the role and underlying mechanism of LncRNA H19 in the pyroptosis of pneumonia has not been elucidated. The purpose of this research was to explore the mechanism by which LncRNA H19 regulates LPS-induced pneumonia in WI-38 cells.METHODS:An LPS induced pneumonia model in WI-38 cells was established. Total RNA extracted from WI-38 cells was analyzed using RT-qPCR, and the total proteins isolated from the WI-38 cells were analyzed using Western blotting. MTT assays, TUNEL staining, bioinformatics, and luciferase reporter assays were subsequently conducted.RESULTS:In the LPS induced pneumonia model, LncRNA H19 silences inhibited LPS-induced WL-38 cell pyroptosis, and LncRNA H19 overexpression promotes LPS-induced WL-38 cell pyroptosis. Also, LncRNA H19 acts as a sponge of miR-22-3p, which targets NLRP3, and NLRP3 attenuates the effect of LncRNA H19 silencing on LPS-induced WL-38 cell pyroptosis.CONCLUSION:Our data demonstrated the roles and potential mechanisms of LncRNA H19 in the regulation of pneumonia cell pyroptosis, indicating that LncRNA H19 is an efficient predictive and curative target for pneumonia.

Abstract Background: Recently, many cases of pneumonia in children with Mycoplasma pneumoniae infection have been shown to have varying degrees of intrabronchial mucus plug formation. The clinical, laboratory, radiological characteristics, and treatment of patients with Mycoplasma infection are analyzed in this study. The risk factors for M. pneumoniae pneumonia (MPP) mucus plug formation in children are explored, and a risk factor scoring system is established.Methods: MPP patients treated with bronchoscopy were retrospectively enrolled in the study from February 2015 to December 2019. The children were divided into a mucus plug group and a control group according to the presence or absence of mucus plug formation. The clinical, laboratory, radiological characteristics, and treatment of the two groups of children were compared. Univariate and multivariate logistic regression models were used to identify the risk factors for MPP mucus plug formation. The receiver operating characteristic (ROC) curve was drawn to evaluate the regression model and establish the MPP mucous plug risk factor scoring system.Results: A univariate analysis showed that the children in the mucous group were older and had a longer fever duration, longer hospital stay, higher fever peak, more cases of wheezing symptoms and allergies, and azithromycin or corticosteroids were administered later. In addition, neutrophil, C-reactive protein (CRP), lactate dehydrogenase (LDH), D-dimer (DD), sputum MP-DNA copy number, and total immunoglobulin A (IgA) levels were higher, while prealbumin (PA) levels were lower. The ROC curve analysis showed that children with MPP had PA ≤144.5 mg/L, had used corticosteroids during the course of the illness of ≥4.5 days, CRP ≥12.27 mg/L, an LDH≥462.65 U/L, and there was a possibility of intra-airway mucus formation. The independent risk factors were scored according to their odds ratio (OR) value. Among the 255 children with MPP, the high-risk group had 44 (83.02%) mucus plugs out of 53; the middle-risk group had 35 (34.3%) mucus plugs out of 102; and the low-risk group had 11 (11%) mucus plugs out of 100.Conclusions: PA levels, timing of corticosteroid use (use in the first few days), CRP levels, and LDH levels were independent risk factors for MPP mucus plug formation. This provides a basis for the early identification of MPP in children combined with mucus plug formation.

Background：There is an increasing attention on miRNAs because of their functional effect on polarization of CD4+ T cells. This study explores the mechanism of miR-493-5p regulating Th9 cell differentiation in allergic asthma. Methods: The allergic airway inflammation is induced by Ovalbumin(OVA)in mice. CD4+ T cells from normal mice are cultured under Th9 cell conditions. IL-9 levels in mice and CD4+T cells are analyzed by RT-qPCR, ELISA, flow cytometry and western blot. The miR-493-5p levels in mice and cells are detected by RT-qPCR. The interaction between FOXO1 and miR-493-5p is predicted by TargetScan and confirmed by dual luciferase assay. The pathological state is evaluated by H&E staining and Lung resistance is measured in the allergic mice treated with miR-493-5p agomiR before stimulation. Results: The miR-493-5p expression in OVA-induced mice decreases significantly, accompanied by a significant upregulation in IL-9, IRF4 and FOXO1 expression and proportion of CD4+Th9 cells. MiR-493-5p mimic inhibits the expression of IL-9, IRF4 and FOXO1 and Th9 cell differentiation, while the inhibitor promotes these effects. MiR-493-5p mimic represses FOXO1 expression through interacting with 3’UTR of FOXO1 mRNA. The rescue experiment proves that miR-493-5p regulates the differentiation of Th9 cell and the expression of IL-9 by targeting FOXO1. In addition, we find that miR-493-5p agomiR treatment inhibits the FOXO1, IL-9 and IRF4 expression, decreases the proportion of CD4+ Th9 cells, alleviates the pathological state of lung tissue and airway hyperreactivity in OVA-induced asthma mice. Conclusions: Our study confirmed that miR-493-5p inhibited Th9 cell differentiation in allergic asthma by targeting FOXO1.

Background Respiratory tract infection is the most common disease in children, and pneumonia is the main disease. Pleural effusion is a common complication of pneumonia. Data show that up to 57% of hospitalized pneumonia patients have pleural effusion [1] . The parapneumonic pleural effusion (PPE) was divided into three categories: uncomplicated parapneumonic effusion (uppe), complex parapneumonic pleural effusion (CPPE) and parapneumonic empyema (PE) [2] .Empyema is a kind of respiratory tract infectious disease with dangerous condition, rapid development and high mortality. Improper treatment can also develop into chronic empyema, and severe complications such as bronchopleural fistula, lung abscess and heart failure may occur. Therefore, timely identification of empyema, active and effective treatment and necessary invasive operation are very important to slow down the disease and reduce the mortality rate. Methods To analyzed the data of children with parapneumonic pleural effusion who were hospitalized in respiratory department of our hospital from January 2010 to December 2018. According to the results of pleural effusion, according to the light classification standard, they were divided into uncomplex parapneumonic effusion (UPPE group) and empyema group (PE group),Logistic regression was used to analyze the risk factors of empyema in children, and receiver operating characteristic (ROC) curve was drawn. Results A total of 63 children were included in the study, including 20 cases in UPPE group and 43 cases in PE group. Univariate analysis showed that children in PE group were younger than those in UPPE group [2.8 (1.1,5.83) vs. 6.54 (2.94,9.0), P < 0.05], clinical manifestations were more prone to shortness of breath (60.46% vs. 8.961, P < 0.05), wheezing (32.55% vs. 5%, P < 0.05), and C-reactive protein (CRP) in laboratory examination was higher [114.76 (54.19160.94) vs. 49.79 (23.68, 84.48), P < 0.05];In cellular immunity, CD3 + was lower (55.31 ± 14.15 vs. 65.76 ± 11.43, P < 0.05), CD3 + CD4 + was lower (27.32 ± 9.58 vs. 34.77 ± 7.99, P < 0.05), CD3-CD19 + was higher (31.61 ± 13.24 vs. 21.98 ± 9.63, P < 0.05), and more susceptible to bacterial infection (44.18% vs. 10%, P < 0.01).Multivariate logistic regression analysis showed that shortness of breath (or = 5.099,95% CI: 1.309-19.866) and elevated CRP (or = 1.013,95% CI: 1.001-1.026) were independent risk factors for empyema. The area under the ROC curve (AUC) of logistic regression model was 0.707 and 0.716, respectively, with moderate diagnostic accuracy. Conclusions The children with parapneumonic pleural effusion with shortness of breath and CRP > 89.41mg/l should be aware of empyema.

Objective: To explore the mechanism that miR-29b and B7H3 regulate the polarization of macrophages and thus affect the differentiation of CD4+T. Methods: 1. PBMC was extracted from peripheral blood mononuclear cells of children with asthma and normal children in the affiliated Children's Hospital of Soochow University, and RNA was extracted and reverse transcribed. The expression of miR-29b and B7H3mRNA was determined by real-time quantitative polymerase chain reaction (Q-PCR). The family history of asthma and history of allergic diseases were collected. 2.THP-1 cells were induced into macrophages, miR-29b interference, miR-29b overexpression and normal control were induced by LV526, LV527 and NC virus infection. After 24 hours of culture, the cells were collected to detect the expression of STAT3 and B7H3 genes and proteins. 3. It was verified that STAT3 was the target gene of miR-29b: after inoculating THP-1 cells and culturing with PMA with final concentration of 50ng/ml for 6 hours, the macrophages without PMA were cultured for 24 hours, then the macrophages infected by LV528, LV529 and NC virus were induced to form miR-29b interference, miR29b overexpression and normal control group. Luciferase analysis was performed at 48 hours to verify that STAT3 was the target gene of miR-29b. STAT3- 3'UTR luciferase reporter gene plasmids were constructed and divided into three groups: "miR-29b+ STAT3- 3'UTR", "miR-29b+STAT3-mut-3'UTR" and "miR-29b+luciferase empty load". 4. Macrophages with different treatments were co-cultured with initial T cells for 3 days. The relative expressions of T-bet, GATA3 and ROR-γt were detected by Q-PCR. Result: 1. The incidence of allergic disease in the acute attack group (68％) was higher than that in the other two groups (34.8％, 33.3％), and the family history of asthma in the normal group (0％) was much lower than that in the other two groups (52％, 60.9％). The difference was statistically significant (P<0.05). 2.The expression of B7H3 in PBMC in acute attack group was higher than that in non-acute attack group and normal group. The expression of miR-29b in PBMC in normal group was significantly higher than that in non-acute attack group and acute attack group (P< 0.0001). The expression of miR-29b in non-acute attack group was significantly higher than that in acute attack group (P= 0.007). 3. After silencing the expression of miR-29b, IL-4Rα, IL-4, IL-5, IL-13 and CD206 of macrophages increased significantly, while IFN- γ decreased, suggesting that miR-29b can promote the polarization of macrophages to M2. 4. The overexpression of miR-29b, STAT3 and B7H3 gene and protein level in macrophages decreased, while the increase of miR-29b, STAT3 and B7H3 gene and protein expression was inhibited. 5. There was a significant positive correlation between the expression of STAT3 and B7H3mRNA in macrophages (r = 0.9737, P< 0.0001). 6. STAT3 is the target gene of miR-29b. 7. Co-culture of macrophages with CD4+T cells can promote the differentiation of primary T cells, namely Th 0 cells, into Th2, and the promoting effect of macrophages with down-regulation of miR-29b is more obvious. Conclusion: The expression of miR-29b in PBMC of children with asthma is lower than that of normal children, while the expression of B7H3 is higher than that of normal children. It is speculated that miR-29b has a protective effect on children with asthma, while B7H3 aggravates the inflammatory response. Down-regulation of miR-29b, in macrophages can promote macrophages to M2 polarization, increase the expression of B7H3 and STAT3 in macrophages, make Th0 cells differentiate into Th2 cells, and aggravate the inflammatory response in patients with asthma.

Background In the past few years, Mycoplasma pneumoniae (Shi et al. Lancet 390:946–958, 2017) infection has been reported more in China. However, there are few studies on the clinical characteristics and prognosis of necrotizing pneumonia (NP) (Griffiths et al. Nature 583:615–619, 2020) caused by different pathogens. Methods A retrospective analysis was performed, including 31 children with a clinical diagnosis of NP in the hospital from January 1, 2013 to January 31, 2020. A total of 11 children with MPNP were included in the observation group and the other 20 children with other pathogens were included in the control group. The clinical manifestations, laboratory data, imaging findings, treatments and outcomes were analyzed. Results The proportion of dyspnea cases was significantly higher in the non- Mycoplasma pneumoniae necrotizing pneumonia (N-MPNP) group than that in the Mycoplasma pneumoniae necrotizing pneumonia (MPNP) group ( P = 0.02).The LDH level of all patients in the MPNP group was higher than the normal value, with a median value of 805.0 U/L, which was significantly higher than those in the N-MPNP group (414.0 [299.9–540.6] U/L; Z = − 2.518; P = 0.012). The white blood cells (WBCs) count of the N-MPNP group was 17.8 (11.1–21.7) × 10 9 /L, which was significantly higher than that of the MPNP group (10.2 [6.3–14.1] × 10 9 /L; P < 0.05). The mean time of pulmonary necrosis in the MPNP group was 20.9 ± 6.9 days, which was higher than that of the N-MPNP group (16.8 ± 6.1 days; t = 3.101; P = 0.004). The incidence of pleural effusion in the N-MPNP group (19 patients, 95%) was significantly higher than that in the MPNP group (six patients, 54.55%) ( P = 0.013). Among them, two patients received bronchoscopy lavage at a maximum four times, and the cases of plastic bronchitis were seen only in the MPNP group (3 cases; P = 0.037).The length of stay was 18 (10–22) days in the MPNP group and 23.5 (13.5–47) days in the N-MPNP group and no significant difference was observed between the two groups (Z = − 1.923, P = − 0.055). Conclusions MP infection is the most common infection in children with NP in the Suzhou area. There is no gender and age difference between MPNP and N-MPNP, but the bacterial infection was mainly observed in the N-MPNP group. Children in the N-MPNP group have more severe clinical symptoms, were more prone to shortness of breath, had a longer hospital stay, and had earlier imaging manifestations of necrosis, whereas children in the MPNP group were more likely to have plastic bronchitis. The level of WBC and LDH and the nature of pleural effusion can be used to identify MPNP and N-MPNP to some extent. The prognosis of MPNP was better than that of N-MPNP. There were no death cases. Pleural thickening, pulmonary fibrosis, and bronchiectasis were the most common sequelae. Compared with N-MPNP, the recovery time of lung imaging in MPNP was shorter.

Abstract Background: There is an increasing attention on microRNAs because of their functional effect on polarization of CD4+ T cells. This study explores the mechanism of miR-493-5p regulating Th9 cell differentiation in allergic asthma.Methods: The allergic airway inflammation is induced by ovalbumin (OVA) in mice. CD4+ T cells from normal mice are cultured under Th9 cell conditions. IL-9 levels in mice and CD4+ T cells are analyzed by RT-qPCR, ELISA, flow cytometry and western blot. The miR-493-5p level in mice and cells is detected by RT-qPCR. The interaction between FOXO1 and miR-493-5p is predicted by TargetScan and confirmed by dual luciferase assay. The pathological state is evaluated by H&E staining and lung resistance is measured in the allergic mice treated with miR-493-5p agomiR before stimulation.Results: The miR-493-5p expression decreases significantly in OVA-induced asthma mice, accompanied by a significant upregulation in the expression of IL-9, IRF4 and FOXO1 and the proportion of CD4+ Th9 cells. MiR-493-5p mimic inhibits the expression of IL-9, IRF4 and FOXO1 and Th9 cell differentiation, while the inhibitor restores these effects. MiR-493-5p mimic represses FOXO1 expression through interacting with 3'UTR of FOXO1 mRNA. The rescue experiment proves that miR-493-5p regulates the differentiation of Th9 cell and the expression of IL-9 by targeting FOXO1. In addition, we find that miR-493-5p agomiR treatment inhibits the FOXO1, IL-9 and IRF4 expression, decreases the proportion of CD4+ Th9 cells, and alleviates the pathological state of lung tissue and airway hyperreactivity in OVA-induced asthma mice.Conclusions: Our study confirmed that miR-493-5p inhibited Th9 cell differentiation in allergic asthma by targeting FOXO1.