Objective:To establish and assess a modified mouse asthma model.Methods:Clean female BALB/c mice were divided into 2 groups(n20,10 for each group).The asthmatic group was sensitized by i.p.and i.n.with OVA aluminum potassium sulfate solution on day 0 and 14.Mice were challenged via the airways with OVA(5% in saline) for 14 days (from day 21 to 35) using ultrasonic nebulization.The control group received saline as the substitution of OVA.Results:The asthmatic group demonstrated the symptoms of acute asthma,such as scratching the head continuously,breathing deeply and fast,staying still,bowing the back,lifting the forelimbs,and so on.There were more leukocytes and eosinophils in the BALF of asthmatic group.In either blood or BALF,the levels of IL-4 and OVA specific IgE were significantly elevated while the IFN-? level was lower in the asthmatic group,as compared with the control group.In asthmatic mice,inflammatory cell increased.The goblet cell hyperplasia,smooth muscle cell proliferation and pulmonary emphysema were much more serious in asthmatic mice than in control mice.Conclusion:This successful modified mouse asthma model may provide a necessary tool in the fundamental research of asthma.

Objective To investigate the etiology,clinical charactieristics,treatment and prognosis in pediatric hemoptysis to provide reference for its diagnosis and treatment to pediatric doctors .Methods The retrospective analysis was performed on the clinical manifestations ,laboratory tests ,imagiological and bronchoscopic manifestations ,treatment methods and outcomes in the children patients with hemoptysis as the chief complaint admitted to our hospital from April 2009 and December 2014 .Results A-mong 102 children cases of hemoptysis ,55 cases were male and 47 cases were female .The median age was (7 .05 ± 1 .2) years old . Mild hemoptysis was in 87 cases(85 .3% ) ,moderate hemoptysis was in 10 cases(9 .8% ) and massive hemoptysis was in 5 cases (4 .9% ) .The relatively exact causes of hemoptysis were found in 88 cases ,in which the lower respiratory tract infection was most common ,the top 3 causes were pneumonia ,pulmonary tuberculosis and bronchitis .In the non-infection causes ,idiopathic pulmonary hemosiderosis(IPH) was most common ,while connective tissue diseases ,pulmonary vascular malformation and airway foreign body were rare .85 cases showed different pathological lesions on chest CT scan .Among the 35 cases underwent the bronchoscopy ,3 ca-ses of bronchial stenosis caused by granulation proliferation was improved after bronchoscopic cryotherapy ,5 cases of active bleed-ing under endoscopic stopped bleeding by topical hemostasis ,among 10 cases of suspected tuberculosis with negative sputum bacte-ria ,acid-fast bacilli was found in 8 cases by bronchofiberscopic lavage .The treatment of pediatric hemoptysis was dominated by the medical etiological and symptomatic therapy .Conclusion Chest CT scan can provide a very important basis for the etiological diag-nosis of hemoptysis and the bronchoscopy examination also has an important value in the diagnosis and treatment of hemoptysis . The etiological and symptomatic treatment is predominat in pediatric hemoptysis ,and the majority have good prognosis .

The oncogenic isoform of the p63 protein, delta NP63, plays an important role in the pathogenesis of many epithelial carcinomas, and emerging evidences suggest that delta NP63 is a promising drug target. However, the functions of delta NP63 in transitional cell carcinoma of bladder (TCCB) are poorly defined. In this study, a delta NP63 shRNA expression vector was transfected into TCCB cell line 5637 and cell cycling, cell proliferation and protein expression were assessed by flow cytometry and 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-dimethyl tetrazolium bromide (MTT) assay, and immunohistochemistry, respectively. The delta NP63 shRNA expression vector was also injected into 5637 cell xenograft tumors in nude mice, and tumor size was measured, tumor tissue morphology was assessed by immunohistopathology and transmission electron microscopy. In the in vitro study, delta NP63 shRNA transfection caused successful delta NP63 gene silencing and resulted in significant arrest of cell cycling and cellular proliferation (p<0.05) as well as cyclin D1 expression. In the nude mouse xenograft model, delta NP63 shRNA greatly inhibited tumor growth, induced tumor cell apoptosis (p<0.05) and resulted in cyclin D1 downregulation. Our data suggest that delta NP63 may play an oncogenic role in TCCB progression through promoting cell survival and proliferation. Intratumoral administration of delta NP63-specific shRNA suppressed tumor delta NP63 expression and cellular proliferation while promoted tumor cellular apoptosis, and therefore inhibited tumor growth and improved survival of xenograft-bearing mice, which was not accompanied by significant signs of systemic toxicity.
Animals ; Carcinoma, Transitional Cell/*genetics/metabolism ; Cell Line, Tumor ; Cell Proliferation ; Cyclin D1/biosynthesis ; Disease Progression ; Female ; Humans ; Mice ; Mice, Nude ; Microscopy, Electron, Transmission ; Models, Biological ; Neoplasm Transplantation ; Trans-Activators/*biosynthesis/*physiology ; Tumor Suppressor Proteins/*biosynthesis/*physiology ; Urinary Bladder Neoplasms/*genetics/metabolism

Objective: To investigate the risk factors for pneumonia in children with Epstein-Barr virus (EBV) infectious mononucleosis(IM). Methods: The clinical data of children with EBV-IM from March 2015 to February 2018 in Children′s Hospital of Chongqing Medical University were retrospectively analyzed.The patients were divided into a pneumonia group and a non-pneumonia group.The difference between 2 groups was analyzed, including gender, age, duration of fever, the maximum temperature in disease duration, the size of liver, the size of spleen, tonsillopharyngitis, cervical lymphadenopathy, edema of the eyelids, white blood cell (WBC), lymphocyte, atypical-lymphocytes, C-reactive protein (CRP), procalcitonin (PCT), alanine transaminase (ALT), glutamic oxalacetic transaminas (AST), lactic dehydrogenase (LDH), cytomegalovirus (CMV) antibody, the titer of mycoplasma (MP) antibody, EBV DNA, length of stay, and hospitalization expenses.The single factor analysis was performed to analyze the above data between 2 groups, and the data with statistical significance were analyzed by the multifactor Logistic regression.The recei-ver operator characteristic (ROC) curve was drawn to evaluate the predicting ability of the indicators for IM combined with pneumonia. Results: Among 923 cases, 133 cases(14.4%) EBV-IM patients were complicated with pneumonia.The findings of single factor analysis indicated that the risk factors were the duration of fever, the size of liver, the size of spleen, and the titer of MP antibody (all P<0.05). The multifactor Logistic regression showed that the duration of fever, the size of liver, the titer of MP antibody were the risk factors for EBV-IM children with pneumonia (P=0.013, 0.028, 0.014). The area under curve (AUC) of the duration of fever was 0.624, and the critical value was 7.5 d (P=0.000); the AUC of the size of liver was 0.590, and the critical value was 2.65 cm (P=0.003). Conclusions The incidence rate of EBV-IM children combined with pneumonia was high.With the presence of the titer of MP antibody ≥1∶160, the duration of fever≥7.5 days, and the size of liver>2.65 cm, it may be independent risk factors for pneumonia in IM children with EBV infection, which requires special attention clinically and earlier chest imageological examination are needed.

Objective:To analyze the patterns of antibacterial agents in Chinese children surveyed by the China multi-center monitoring network for the application of antibacterial agents in children and neonates in 2019 by using World Health Organization (WHO) Access, Watch, Reserve and Not-recommended (AWaRe) and typical anatomical/therapeutic/chemical (ATC) in this study.Methods:The cross-sectional method was adopted.A multi-center cross-sectional survey was conducted on one day from September to December 2019.The information of all inpatients taking antibiotics was uploaded to the network-based data collection system (https: //garpec-31.mobilemd.cn/login.aspx? relogin=true). This study covered 13 hospitals from 10 provinces and cities in China.All hospitalized children in the Respiratory Department, Infectious Disease Department, General Surgery Department, Pediatric Intensive Care Units, Neonatal Intensive Care Units and Neonatology joined in this survey.The clinically used antibacterial agents were classified by AWaRe and ATC, and the AWaRe and ATC distributions of antibacterial agents prescribed for Chinese children and neonates were described.Results:Of the 2 644 antibiotic prescriptions included from 13 hospitals, 2 134 (80.71%) were for children and 510 (19.29%) were for neonates.Of all antibiotic prescriptions, there were 368 (13.92%) Access antibiotics prescriptions, 1 973 (74.62%) Watch prescriptions, 60 (2.27%) Reserve prescriptions and 243 (9.19%) Not-recommended prescriptions.The top-five antibiotics prescribed for children and neonates were third-generation cephalosporins (1 056, 39.94%), macrolides (492, 18.61%), carbapenems (275, 10.40%), beta lactam-beta lactamase inhibitors (246, 9.30%), and second-generation cephalosporins (136, 5.14%). The use ratios of Access, Watch, Reserve and Not-recommended antibiotics in each center ranged from 0 to 30.00%, 36.67% to 97.20%, 0 to 17.02% and 0 to 33.33%, respectively.In 1 360 antibiotic prescriptions for children and neonates with pneumonia, there were 152 (11.18%) Access antibiotics, 1 051 (77.28%) Watch antibiotics, 37 (2.72%) Reserve antibiotics, and 120 (8.82%) Not-recommended antibiotics.The top-five antibiotics prescribed for children with pneumonia were third-generation cephalosporins (522, 38.38%), macrolides (388, 28.53%), beta lactam-beta lactamase inhibitors (141, 10.37%), carbapenems (117, 8.6%) and penicillins (49, 3.60%).Conclusions:Watch antibiotics and broad spectrum antibiotics such as third-generation cephalosporins and macrolides prone to induce resistance are the main antibacterial agents used in Chinese children and neonates with pneumonia.Broad-spectrum antibiotics may be overused in Chinese children and neonates.

Objective To investigate the effect of Brahma-related gene 1 (Brg1) on mucus hypersecretion in the airway of asthmatic mice and explore the mechanism. Methods Female C57bl/6 mice aged 6-8 weeks were randomized into wild-type control group, wild-type asthma group, Brg1-/-group with Brg1 gene knockdown in type II alveolar epithelial cells, and Brg1-/-+asthma group (n=10). The mice in asthma group and Brg1-/-+asthma group were sensitized with ovalbumin (OVA) to establish asthmatic models. PAS staining was used to determine the number of goblet cells and mucus secretion in the airway. Real-time PCR was used to detect the expression of MUC5AC mRNA in the lung tissues. The levels of mucin MUC5AC and interleukin-13 (IL-13) in the bronchoalveolar lavage fluid (BALF) were detected with ELISA and immunohistochemistry, and the expressions of STAT6 and p-STAT6 in the lung tissue were detected using Western blotting. Results Compared with the control mice, wild-type asthmatic mice showed obvious mucus hypersecretion and increased MUC5AC mRNA in the airway with significantly increased IL-13 and MUC5AC levels in the BALF and activation of p-STAT6 in the lung tissues (P<0.05). In the transgenic mice with Brg1 gene knockdown, airway mucus secretion and MUC5AC mRNA expression was significantly reduced following OVA challenge compared with those in the wild-type asthmatic mice; IL-13 and MUC5AC levels in the BALF and p-STAT6 expression in the lung tissues were also significantly decreased in the transgenic mice (P<0.05). Conclusion Brg1 gene knockdown in type II alveolar epithelial cells alleviates OVA-induced airway mucus hypersecretion and reduces the expression of MUC5AC in C57bl/6 mice possibly by inhibiting STAT6 activation, suggesting the role of Brg1 in promoting asthmatic airway mucus hypersecretion.

Objective To investigate the effect of Brahma-related gene 1 (Brg1) on mucus hypersecretion in the airway of asthmatic mice and explore the mechanism. Methods Female C57bl/6 mice aged 6-8 weeks were randomized into wild-type control group, wild-type asthma group, Brg1-/-group with Brg1 gene knockdown in type II alveolar epithelial cells, and Brg1-/-+asthma group (n=10). The mice in asthma group and Brg1-/-+asthma group were sensitized with ovalbumin (OVA) to establish asthmatic models. PAS staining was used to determine the number of goblet cells and mucus secretion in the airway. Real-time PCR was used to detect the expression of MUC5AC mRNA in the lung tissues. The levels of mucin MUC5AC and interleukin-13 (IL-13) in the bronchoalveolar lavage fluid (BALF) were detected with ELISA and immunohistochemistry, and the expressions of STAT6 and p-STAT6 in the lung tissue were detected using Western blotting. Results Compared with the control mice, wild-type asthmatic mice showed obvious mucus hypersecretion and increased MUC5AC mRNA in the airway with significantly increased IL-13 and MUC5AC levels in the BALF and activation of p-STAT6 in the lung tissues (P<0.05). In the transgenic mice with Brg1 gene knockdown, airway mucus secretion and MUC5AC mRNA expression was significantly reduced following OVA challenge compared with those in the wild-type asthmatic mice; IL-13 and MUC5AC levels in the BALF and p-STAT6 expression in the lung tissues were also significantly decreased in the transgenic mice (P<0.05). Conclusion Brg1 gene knockdown in type II alveolar epithelial cells alleviates OVA-induced airway mucus hypersecretion and reduces the expression of MUC5AC in C57bl/6 mice possibly by inhibiting STAT6 activation, suggesting the role of Brg1 in promoting asthmatic airway mucus hypersecretion.

Objective:To analyze the cystic fibrosis transmembrane conductance regulator (CFTR) gene variations and phenotypes in 7 Chinese children.Methods:In this retrospective study, the data of 7 children with CFTR gene variations admitted to Children′s Hospital of Chongqing Medical University from December 2013 to October 2020 were extracted. The general information, clinical manifestations, gene variations, diagnosis and treatment were summarized.Results:Among the 7 children, 2 were males and 5 were females, aged 5.2(0.5-11.3) years. Main clinical manifestations included malnutrition (5 cases), recurrent respiratory infection (4 cases), bronchiectasis (3 cases), steatorrhea (3 cases), vomiting in infancy (2 cases), liver cirrhosis (2 cases), meconium ileus (1 case), metabolic alkalosis and hypochloremia (1 case). A total of 15 variations were found by whole exon sequencing and Sanger sequencing, among which 3 were newly discovered, and 7 were missense mutations. Four children were diagnosed as CF, and the other 3 were diagnosed as CFTR related disease (CFTR-RD). Compared with CF patients, the pancreatic insufficiency and typical CF lung disease were relatively mild in CFTR-RD patients. After treatment, 6 children were clinically improved, while the rest one withdrew treatment due to critical pulmonary infection and disturbance of water-electrolyte metabolism.Conclusions:The loci and phenotypes of CFTR gene variants vary hugely and the pathogenicity of some variations are not clear. Whole exon sequencing can facilitate the identification of CF-and CFTR-RD-causing variaions. For the cases not compatible with CF, CFTR-RD should be considered and evaluated by timely gene detection, so as to carry out appropriate long term management.