N-Acetyl-L-Cysteine Protects Airway Epithelial Cells during Respiratory Syncytial Virus Infection against Mucin Synthesis, Oxidative Stress, and Inflammatory Response and Inhibits HSPA6 Expression.

Objective. Respiratory syncytial virus (RSV) infection is an important cause of hospitalization of children worldwide, leading to significant morbidity and mortality. RSV infection leads to increasing inflammatory and apoptosis events in the airway epithelium through mechanisms involving ROS generation. The antioxidant N-acetyl-L-cysteine (NAC) has been shown to inhibit influenza virus replication and to reduce the secretion of inflammatory and apoptotic mediators during virus infection. The study aims to investigate the effects of NAC on human bronchial epithelial cells BEAS-2B and HSPA6 expression during RSV infection. Methods. CCK-8 assays were performed to evaluate cell survival. The production of proinflammatory factors, TNF-α, IL-6, IL-1ß, IL-18, and MUC5AC was examined by quantitative real-time PCR and ELISA. Oxidative stress was determined by reactive oxygen species (ROS), superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione (GSH)/glutathione disulfide (GSSG) ratio. Immunoblotting analysis of epidermal growth factor receptor (EGFR) and its phosphorylation was performed. The antiviral effect of NAC was assessed by determining viral titers using plaque assay. Results. RSV infection reduced cell survival, promoted the release of proinflammatory factors, increased the ROS production and MDA concentration, and diminished the SOD activity and GSH/GSSG ratio, all which were attenuated by NAC treatment. Accordingly, NAC treatment inhibited the activation of EGFR and MUC5AC in BEAS-2B cells with RSV infection. Furthermore, NAC administration resulted in a marked decrease in RSV-induced HSPA6 expression in BEAS-2B cells. Concomitantly, EPB treatment led to an evident inhibition of RSV fusion gene and viral replication in RSV-infected BEAS-2B cells. Conclusion. This work supports the use of NAC to exert antimucin synthesis, anti-inflammatory, antioxidant, and antiviral effects on airway epithelium during RSV infection.

[Liquid chromatography-mass spectrometry-based metabolomics study of the efficacy of Chinese medicine asthma-relieving decoction on respiratory syncytial virus infection].

Respiratory syncytial virus (RSV) can cause lower respiratory tract infections, such as bronchiolitis in infants. In China, traditional asthma-relieving medicine has numerous clinical applications in the treatment of RSV infections. However, due to the complexity of the traditional Chinese medicine system, its therapeutic mechanism and main pharmacological components remain unclear. Metabolomics can be used to analyze the efficacy of traditional Chinese medicine to provide modern scientific evidence for such treatments. In this study, an animal model experiment was performed with seven groups of three-week-old rats. The model group and five intervention groups were inoculated nasally with RSV for three consecutive days, and the normal group was treated with the same amount of saline for three consecutive days under the same conditions. In parallel, the five intervention groups were treated separately with the following via intragastric administration for seven consecutive days: asthma-relieving traditional Chinese medicine decoction, its three constituent agents (ascending (xuan) therapy, descending (jiang) therapy, pyretic clearing (qing) therapy), and ribavirin. Both normal group and RSV model group were administered with normal saline via intragastric administration as controls for seven consecutive days. The fundus plasma of rats in each group was collected on day 0, day 3, and day 7. Liquid chromatography-mass spectrometry-based untargeted metabolomics analysis was performed to investigate the changes in the metabolome after RSV infection, the effects of the asthma-relieving decoction on the regulation of metabolites related to RSV infection, and the primary source of efficacy. The detected metabolite ions were corrected using internal standards. Multivariate analysis of ions with an RSD value of less than 30% in quality control (QC) samples was used to construct principal component analysis models to monitor the overall metabolic changes of each group. The results showed that, during RSV infection and treatment, the asthma-relieving decoction and the positive control ribavirin had similar effects on the overall metabolic regulation of RSV-infected rats. Among the three asthma-relieving decoction constituent agents, the ascending (xuan) therapy agents which was composed of ephedra and ginkgo had a closer metabolic regulation effect with asthma-relieving decoction, and might be the main source of pharmacological efficacy. Based on the retention time, m/z value and tandem mass spectra in the database established by our laboratory, a total of 150 metabolites were identified. Paired t-tests were performed using data of the identified metabolites before and after RSV infection in each group, and it was found that 83 metabolite levels significantly changed after RSV infection, indicating that RSV infection could lead to disorders of multiple metabolic pathways in rats. The altered pathways included aminoacyl-tRNA biosynthesis, phenylalanine, tyrosine, and tryptophan biosynthesis, primary bile acid biosynthesis, phenylalanine metabolism and sphingomyelin metabolism. On the third day, the asthma-relieving decoction had regulatory effects on several metabolites such as bile acids, amino acids, organic acids, lipids, etc. Among the three asthma-relieving decoction constituent agents, the ascending (xuan) therapy agents had more similar effects on the regulation of metabolites with the asthma-relieving decoction. On the other hand, the descending (jiang) therapy agents and pyretic clearing (qing) therapy agents down-regulated the abnormal increase in acylcarnitine caused by the RSV infection. Additionally, both asthma-relieving decoction and its constituent agents could maintain the stability of the immune system and metabolism of the intestinal flora in rats. This study used metabolomics to evaluate the efficacy of an asthma-relieving decoction and demonstrate the metabolites and the corresponding changes after asthma-relieving decoction-based treatment. It provides theoretical support for research on the therapeutic mechanism and active ingredients of asthma-relieving decoction.

Improving Cycling Stability and Rate Capability of High-Voltage LiCoO2 Through an Integration of Lattice Doping and Nanoscale Coating.

High-voltage LiCoO2 has attracted much interest owing to the high specific energy density. But the poor cycling performance and inferior rate capacity of LiCoO2 at a high voltage (≥4.5 V) has restricted the practical applications. Herein, we propose to improve the electrochemical performances of LiCoO2 at high voltage through a synergy of Al-doping and Li2TiO3-coating. In compared to bare LiCoO2, Al-doped LiCoO2 and Li2TiO3-coated LiCoO2, the cycle performance, the rate capability and the polarization of Al-doped and Li2TiO3-coated LiCoO2 shows a larger improvement, which can be attributed to the synergic effects of Al-doping and Li2TiO3-coating. Firstly, Al doping expands the interlayer spacing which decreases the Li-ion diffusion barrier and enhances the coefficient of Li-ion diffusion. This benefits to the rate capability. Secondly, Al doping enhances the layered structure stability due to the larger Al-O bonding energy (ΔH298 Kt (Al-O) = 512 kJmol-1) than that of Co-O (ΔH298 Kt (Co-O) = 368 kJmol-1). Thirdly, the coating layer of Li2TiO3 mitigates the surface side reactions and further enhances the cycling performance. Moreover, the coating layer of Li2TiO3 as a Li+-conductor is also favorable to the Li+ diffusion and the rate capability. This synergic strategy can also be extended to the modification of other cathode materials.

[Levels of lymphocyte subsets, immunoglobulins, and complement C3 and C4 in children with hand-foot-mouth disease].

OBJECTIVE: To study the clinical value of lymphocyte subsets, immunoglobulins, and complement C3 and C4 in the evaluation of immune status in children with hand-foot-mouth disease (HFMD). METHODS: A total of 282 children with HFMD were enrolled as the HFMD group, and 130 healthy children were enrolled as the healthy control group. The percentages of peripheral CD3+, CD4+, and CD8+ T lymphocytes, CD19+ B lymphocytes, and CD56+ natural killer cells were measured. The CD4+/CD8+ ratio was calculated. The levels of immunoglobulin A (IgA), immunoglobulin M (IgM), immunoglobulin G (IgG), and complement C3 and C4 were measured. RESULTS: The multivariate analysis showed that compared with the healthy control group, the HFMD group had significantly lower percentages of CD3+, CD4+, and CD8+ T lymphocytes and levels of complement C3 and C4 (P<0.05), as well as significantly higher percentage of CD56+ natural killer cells and level of IgG (P<0.05). The individual effect analysis showed that the children aged 0-3 years in the HFMD group had a significantly higher CD4+/CD8+ ratio than the healthy control group (P<0.05); boys aged 0-3 and ≥3 years in the HFMD group had a significantly higher level of IgM than the healthy control group (P<0.05); boys aged ≥3 years and girls aged 0-3 years in the HFMD group had a significantly lower level of IgA than the healthy control group (P<0.05). CONCLUSIONS: Cellular and humoral immunity disorders are observed in children with HFMD. The monitoring of lymphocyte subsets and immunoglobulin levels can provide a laboratory basis for immune status assessment in children with HFMD.

[Status of antibiotic use in hospitalized children with community-acquired pneumonia in multiple regions of China].

OBJECTIVE: To investigate the use of antibiotics in children with community-acquired pneumonia (CAP) in multiple regions of China, and to provide a reference for CAP standard treatment and rational antibiotic use in children. METHODS: The medical data of 1 383 children with CAP who were hospitalized in the department of pediatrics in 10 grade A tertiary hospitals from 9 cities between April 14, 2014 and January 1, 2016 were reviewed, to analyze the status of antibiotic use in hospitalized children in North China, Northeast China, East China, and South China. RESULTS: The overall rate of antibiotic use in children with CAP was 89.08%, with 88.7% in North China, 95.5% in Northeast China, 83.3% in East China, and 86.6% in South China. The main types of antibiotics used were cephalosporins, macrolides, compound preparations of ß-lactam antibiotics, polyphosphoric broad-spectrum antibiotics and other ß-lactam antibiotics. The selection of antibiotics was generally rational, but antibiotics were still used in some patients with viral infection alone or a combined use of ≥2 kinds of antibiotics were noted in some patients with infection caused by one kind of pathogen. Irrational antibiotic use was observed in 131 children (10.63%). CONCLUSIONS: There are high rates of antibiotic use and irrational use of antibiotics among children with CAP. Standard management of antibiotic use in children with CAP should be strengthened.

Serum metabolomics study and eicosanoid analysis of childhood atopic dermatitis based on liquid chromatography-mass spectrometry.

Atopic dermatitis (AD) is the most common inflammatory skin disease in children. In the study, ultra high performance liquid chromatography-mass spectrometry was used to investigate serum metabolic abnormalities of AD children. Two batch fasting sera were collected from AD children and healthy control; one of them was for nontargeted metabolomics analysis, the other for targeted eicosanoids analysis. AD children were divided into high immunoglobulin E (IgE) group and normal IgE group. On the basis of the two analysis approaches, it was found that the differential metabolites of AD, leukotriene B4, prostaglandins, conjugated bile acids, etc., were associated with inflammatory response and bile acids metabolism. Carnitines, free fatty acids, lactic acid, etc., increased in the AD group with high IgE, which revealed energy metabolism disorder. Amino acid metabolic abnormalities and increased levels of Cytochrome P450 epoxygenase metabolites were found in the AD group with normal IgE. The results provided a new perspective to understand the mechanism and find potential biomarkers of AD and may provide a new reference for personalized treatment.

[Effect of bronchoalveolar lavage on the heart rhythm and conduction of the children with severe pneumonia].

OBJECTIVE: To observe the effect of bronchoalveolar lavage on the heart rhythm and conduction of children with severe pneumonia through monitoring the electrocardiogram change of different step of the bronchoalveolar lavage, for proving the safety of the operation of bronchoalveolar lavage from the perspective of cardiac electrophysiology. METHOD: From July 2011 to March 2012, 30 patients who were hospitalized in pneumology department of Dalian Children's Hospital and met the inclusion criteria and therapeutic indications of bronchoalveolar lavage were chosen. They were 3 to 12 years old, the average age was 5.3 years, including 17 boys and 13 girls, the ratio of boys and girls is 1.3: 1. Continuous sampling the electrocardiogram before and during the process including anesthesia, entering into glottis, lavage, aspiration, and revive, and recording the heart rate, rhythm amplitude and width of P wave, the PR interval, the form and width of QRS complex were also measured. The recorded data were analyzed and statistical analysis to reflect the change of the cardiac electrophysiology. RESULT: The incidence of heart rate increase was 100.0%, 26 (86.7%) patients began to emerge after anesthesia, the rest of the patients also developed heart rate increase after the start of bronchoscopic operation. All patients had sinus tachycardia, and were most obvious in the progress of lavage and revive. In the process of entering into glottis, lavage, aspiration, 13 (43.3%) patients had arrhythmia episodes. Types of arrhythmia included sinus bradycardia, atrioventricular block and premature beat. Incidences of intraoperative arrhythmia compared with the pre- and post-operation were all statistically significantly different (P = 0.00). The most common arrhythmia were premature beat, in 17 of the 30 cases there were premature beat including 9 cases with atrial premature beats and 8 cases ventricular premature contraction. Two patients had III° atrioventricular block accompanied by serious sinus bradycardia. All kinds of arrhythmias except sinus tachycardia disappeared after the operation was ended. Five patients (16.7%) had PR interval prolongation. Five patients (16.7%) had incomplete right bundle branch block (IRBBB) . Incidences of IRBBB compared with the pre-operation and post-operation were all not significantly different [13.3% (n = 4) vs. 0(n = 0) and 3.3% (n = 1), all P > 0.05]. Different operating progress made no significant difference in the measurement value of electrocardiogram[13.3% (n = 4) vs. 0(n = 0) and 3.3% (n = 1), all P > 0.05], but showed the most notable effect on heart rate. CONCLUSION: Bronchoalveolar lavage can influence the heart rhythm and conduction, but most of the influence with pathological significance are transient. Cardiac electrophysiological changes were the most obvious in endotracheal operation with the risk of malignant arrhythmia but the risk is low, the bronchoalveolar lavage technique is safe. The contingency plans for dealing with all kinds of adverse reactions must be ready before the operations of bronchoalveolar lavage. During the procedure, the patient's ECG changes should be closely monitored to reduce the incidence of adverse reactions.

[Application of combined esophageal multichannel intraluminal impedance-pH monitoring in infants and children with gastroesophageal reflux disease].

OBJECTIVE: This study aimed to evaluate the diagnostic value of a 24-hour esophageal combined multichannel intraluminal impedance pH (24 h our MII-pH) monitoring in children with gastroesophageal reflux (GER) disease and recurrent pneumonia. METHODS: A total of 17 cases with a suspected diagnosis of GER disease children with unexplained recurrent pneumonia underwent a 24-hour MII-pH monitoring to analysis of the nature and characteristics of reflux. RESULTS: In the 17 cases of GER disease with recurrent pneumonia, 11 (65%) were confirmed positively by a 24-hours of MII-pH monitoring. A total of 853 reflux cycle were detected, of which 65.3% were acid refluxes predominantly occurring within 2 hours after meal, and 71.6% were mixed refluxes. Refluxes occurred most frequently in the supine position, followed by the lateral position and the prone position. Distal reflux was the predominant form of reflux. According to the Biox-Ochoa classification, 73% of cases (8/11) were severe GER. CONCLUSIONS: MII-pH monitoring may effectively characterize refluxate properties and thereby increase the detection rate of GER disease.

A cross-sectional study of the clinical characteristics of hospitalized children with community-acquired pneumonia in eight eastern cities in China.

BACKGROUND: Community-acquired pneumonia in children is common in China. To understand current clinical characteristics and practice, we conducted a cross-sectional study to analyze quality of care on childhood pneumonia in eight eastern cities in China. METHODS: Consecutive hospital records between January 1, 2010 and December 31, 2010 were collected from 13 traditional Chinese medicine (TCM) and western medicine (WM) hospitals in February, May, August, and November (25 cases per season, 100 cases over the year), respectively. A predesigned case report form was used to extract data from the hospital medical records. RESULTS: A total of 1298 cases were collected and analyzed. Symptoms and signs upon admission at TCM and WM hospitals were cough (99.3% vs. 98.6%), rales (84.8% vs. 75.0%), phlegm (83.3% vs. 49.1%), and fever (74.9% vs. 84.0%) in frequency. Patients admitted to WM hospitals had symptoms and signs for a longer period prior to admission than patients admitted to TCM hospitals. Testing to identify etiologic agents was performed in 1140 cases (88.4%). Intravenous antibiotics were administered in 99.3% (595/598) of cases in TCM hospitals and in 98.6% (699/700) of cases in WM hospitals. Besides, Chinese herbal extract injection was used more frequently in TCM hospitals (491 cases, 82.1%) than in WM hospitals (212 cases, 30.3%) (p < 0.01). At discharge, 818 cases (63.0%) were clinically cured, with a significant difference between the cure rates in TCM (87.6%) and WM hospitals (42.0%) (OR = 9.8, 95% confidence interval (CI): 7.3 ~ 12.9, p < 0.01). Pathogen and previous medical history were more likely associated with the disappearance of rales (OR = 7.2, 95% CI: 4.8 ~ 10.9). Adverse effects were not reported from the medical records. CONCLUSIONS: Intravenous use of antibiotics is highly prevalent in children with community-acquired pneumonia regardless of aetiology. There was difference between TCM and WM hospitals with regard to symptom profile and the use of antibiotics. Intravenous use of herbal injection was higher in TCM hospitals than in WM hospitals. Most of the cases were diagnosed based on clinical signs and symptoms without sufficient confirmation of aetiology. Audit of current practice is urgently needed to improve care.

[Distribution and drug resistance of the isolated bacteria from children with acute respiratory infection].

OBJECTIVE: To study the distribution and drug resistance of the isolated bacteria from children with acute respiratory infection (ARI) in Dalian. METHODS: Between January 2006 and February 2007, 930 children with ARI were enrolled, including 364 with acute upper respiratory infection (AURI), and 566 with acute lower respiratory infection (ALRI). The AURI children, who did not receive antimicrobial agent treatment or received oral antimicrobial agents 1-2 times, had bacterial cultures of pharyngeal swab. The ALRI children, who received intravenous antibacterial agents more than 3 days, had bacterial cultures of sputum and bronchoalveolar lavage fluid (BALF). Isolated bacteria were identified by the ATB system (Bio-Merieux, France). Antimicrobial susceptibility testing was carried out by means of Kirby-bauer. RESULTS: A total of 404 isolates (43.4%) were identified. Haemophilus influenzae, Haemophilus parainfluenzae and Streptococcus pneumoniae accounted for 22.5%, 12.1% and 7.4% respectively. In the isolates from AURI, Haemophilus influenzae, Haemophilus parainfluenzae and Streptococcus pneumoniae accounted for 43.9%, 22.0% and 9.1% respectively; Escherichia coli, Klebsiella pneumonia and Nonfermenters accounted for 4.5%, 8.3% and 3.0% respectively. In the isolates from ALRI, Haemophilus influenzae, Haemophilus parainfluenzae and Streptococcus pneumoniae accounted for 12.1%, 7.4% and 6.6% respectively; Escherichia coli, Klebsiella pneumoniae and Nonfermenters accounted for 16.9%, 13.2% and 21.8% respectively. The resistant rates of Haemophilus to ampicillin and TMP-SMZ were 29.3% and 32.9% respectively, and to amoxicillin-clavulanic acid, cefalotin, cefaclor, cefuroxime and cefotaxime were 12.1%, 10.0%, 10.0%, 11.4% and 5.7%, respectively. The resistant rate of Haemophilus to ampicillin, amoxicillin-clavulanic acid, cefaclor, tetracycine and TMP-SMZ in the ALRI group were significantly higher than that in the AURI group (P<0.05 or 0.01). CONCLUSIONS: In Dalian, Haemophilus was the main isolate of children with ARI. The distribution of bacteria was different between ALRI and AURI. In ALRI, Gram-negative bacilli were in a higher proportion, and the resistant rates of Haemophilus influenzae and Haemophilus parainfluenzae to ampicillin, amoxicillin-clavulanic acid and cefaclor were higher.

[Dingchuantang decoction restores the imbalance of TH2/TH1 in mice infected by respiratory syncytial virus].

OBJECTIVE: Dingchuantang is a traditional Chinese medicine decoction which has the Qing, Xuan and Jiang components which have the ability to dispel (Xuan), depress (Jiang) and heat clear or febrigugal (Qing). It has been used for asthma and respiratory syncytial virus (RSV) infection. However, the therapeutic mechanisms of the decoction have not been identified. To understand the mechanism of the anti-RSV property of Dingchuantang, this study investigated the effects of Dingchuantang and its three components on the levels of interleukin-4 (IL-4) and interferon-gamma (gamma-IFN) in mice infected by RSV. METHODS: One hundred and twenty mice were randomly assigned into Normal, Infected, Dingchuantang-treated, Qing component-treated, Xuan component-treated and Jiang component-treated groups (n=20 each). The RSV infection model was prepared in the rats from the last 5 groups. The 4 treatment groups were administered with Dingchuantang, Qing component, Xuan component and Jiang component by gastric lavage respectively 2 hrs after RSV innoculation. The contents of RSV in the lungs were tested by reverse transcription polymerase chain reaction (RT-PCR). The levels of IL-4 and gamma-IFN in broncho-alveolar lavage fluid (BALF) were measured by enzyme linked immunosorbent assay (ELISA). RESULTS: The contents of RSV of lungs in the Dingchuantang-treated group, and Qing and Xuan component-treated groups were significantly lower than those in the untreated Infected group. There was no significant difference between the Jiang component-treated and the Infected group. Dingchuantang treatment decreased the IL-4 level and increased the gamma-IFN level. The Xuan and Jiang component treatment decreased the IL-4 level but had no effect on the gamma-IFN level, while the Qing component treatment increased the gamma-IFN level but had no effect on the IL-4 level. CONCLUSIONS: Dingchuantang can inhibit RSV reproduction and this effect works by Qing and Xuan components. Dingchuantang can also restore the imbalance TH2/TH1 by depressing IL-4 levels, worked by Xuan and Jiang components, and increasing gamma-IFN levels, worked by Qing component.

[Effect of intrauterine acute ischemic-hypoxia on the expression of lung SP-A and SP-B in neonatal rats].

OBJECTIVE: Neonatal asphyxia is one of the main causes for the acute respiratory distress syndrome (ARDS) in full-term newborns. Now it is believed that the reduced amount and abnormal function of pulmonary surfactant due to various causes is a major factor leading to acute lung injury. This study aimed at using an intrauterine acute ischemic-hypoxia rat model and investigating the effect of intrauterine acute ischemic-hypoxia on the expression of surfactant protein A (SP-A) and surfactant protein B (SP-B) in neonatal rat lungs. METHODS: The rat model of acute intrauterine ischemic-hypoxia was established by ligating the unilateral uterine horn vessels of Wistar rats at the 21st gestational day. While the rat pups from the other side of the uterus, of which the uterine horn vessel was not ligated, were the sham-operation group. Rat pups were delivered by cesarean section at the 20, 30 and 40 min following the ischemic-hypoxia insult. The rat pups delivered by cesarean section from the gestation of 21 days were the normal control group. There were 42 rat pups and 6 pups in each group in this study. The distribution of SP-B protein in the neonatal rat lungs of different period of ischemia was examined by using SABC method. The average gray value of SP-B staining in type II alveolar epithelial cells were measured by Universal Imaging Porporation with Meta Morph software. The reverse transcription polymerase chain reaction (RT-PCR) was performed to quantitate the expression of SP-A and SP-B mRNA. RESULTS: Following the intrauterine acute ischemic-hypoxia, the numbers of type II alveolar epithelial cells with the positive SP-B staining were markedly declined. The average gray values at the 20, 30 and 40 min after the ischemia were 78.89 +/- 1.08, 79.69 +/- 0.13 and 80.00 +/- 0.63, respectively, which increased significantly compared with the normal control group (76.13 +/- 0.43, P < 0.01). The expression of SP-A and SP-B mRNA was weak following the ischemic-hypoxia insult. The relative amounts of SP-A (1.16 +/- 0.06, 1.14 +/- 0.01 and 1.13 +/- 0.04, respectively) and SP-B (0.81 +/- 0.02, 0.78 +/- 0.02 and 0.79 +/- 0.04, respectively) at the 20, 30 and 40 min after the ischemia were reduced significantly compared with controls (1.27 +/- 0.09 and 0.89 +/- 0.06, respectively, P < 0.05 and < 0.01) and reduced gradually following the prolongation of the insult. There were no significant differences (P > 0.05) between the normal and sham operation control groups on the expressions of SP-B protein as well as the SP-A and SP-B mRNA. CONCLUSION: The reduced synthesis of SP-B protein and the reduced expression of SP-A and SP-B mRNA might be caused by intrauterine acute ischemic-hypoxia, which may support theoretically the early application of pulmonary surfactant including SP-A and SP-B for treating the lung injuries of asphyxia in newborns.