Objective To provide reference for anti-infection drugs in Zunyi area by analyzing the etiological characteristics of children with severe pneumonia.Methods The sputum, throat swabs and serum of children with severe pneumonia in pediatric intensive care unit of our hospital from January 2014 to December 2015 were collected in this study.The pathogen species which caused severe pneumonia were detected and identified by the method of pathogen culture,and typical pathogens were detected by RT-PCR and indirect immunofluorescence.Results A total of 337 children with severe pneumonia were included,the pathogen positive rate was 86.65%(292/337).The rate of viral infection(37.32%)was the highest,followed by bacterial infection accounting for 28.42%,then the mixed infection accounting for 27.74%,and the mycoplasma pneumoniae infection accounting for 6.50%.The respiratory syncytial virus type B accounting for 28.44% was the most common in viral infection,and there were significant differences in age distribution(P<0.05),the children under 3-years-old,especially the infants under 1-year-old had the highest susceptibility.Viral infection had certain seasonality,compared with spring and winter,autumn and winter(November to April) had higher viral detection rate and the difference was statistically significant(x2=29.28,P=0.001).The escherichia coli was the most common in bacterial infection,accounting for 21.69%.Klebsiella pneumoniae,Escherichia coli were more common in neonates and infants under 1-year-old,Haemophilus influenzae occured in 1~3 years old children,Streptococcus pneumoniae infection could occur in any age.Conclusion Viral infection is the most common pathogen in children with severe pneumonia in Zunyi area followed by bacterial infection,which is more common in children under 1-year-old,and with the high incidence in autumn and winter.Bacterial infection is more common in children over 3 years of age.Bacterial and viral mixed infection is common in children under 3 years of age,especially in children under 1-year-old.

Objective To inquire into the molecular epidemiology of enterovirus 71 ( EV71 ) in our region by analyzing the whole genome characteristics and genetic evolution of EV 71 strains isolated from Guizhou area. Methods The throat swabs samples of hospitalized children with hand,foot and mouth disease in Guizhou province from 2013 to 2015 were collected,the virus nucleic acid were extracted,then the whole genome of virus were piecewise amplified by reverse transcription polymerase chain reaction( RT-PCR) and sequenced. Sequencing results were edited and spliced by DNAMAN8. 0 software,then the viral genome se-quences were compared with genome sequences of other EV71 strains in the genebank by Blastn,the phyloge-netic tree was constructed by the Neighbor-Joining method in MEGA5. 2 software. Results The whole ge-nome sequences of 17 EV71 strains were successfully isolated and amplified,the whole genome length of 17 EV71 isolates was 7405 base pair,encoded about 2193 amino acids. The 17 isolates were divided into ten species of amino acid sequences by 12 differences of amino acid among the strains,different sequences and clinical types had not shown regularity and correlation. The nucleotide homology in VP1 region,5′untranslat-ed region(5′UTR) and 3′untranslated region (3′UTR) were high among 17 EV71 isolates. The results that the whole genome of 17 EV71 isolates was compared with representative strains of EV71 A,B,C genotype and coxsakievirus A 16 ( CA16) showed that 17 EV71 isolates had higher homology with EV71 C4a sub-type,95. 3%-98. 1%,and the lowest homology with CA16. The phylogenetic tree was constructed based on nucleotide sequence of the whole genome,VP1 region and 5′untranslated region of 17 isolates showed that the 17 isolates were clustered into one cluster,and were clustered in the same branch with C4a isoforms,the phy-logenetic relationships among different regions were different. Conclusion The popular genotype of EV71 strains in Guizhou area for 2013-2015 was C4a subtype,consistenting with the genotype of popular EV71 in other regions of China. EV71 strains hasn′t the antigen transformation and input of a new subtype temporari-ly,but exist nucleotide and amino acid changes,so need be chronically and dynamically monitored. There is no correlation between the amino acid sequence difference of 17 EV71 isolates and the state of an illness.

OBJECTIVE To investigate the preventive effect of cucurbitacin B （CB） on sepsis-induced acute lung injury （ALI） and its mechanism. METHODS The mice were divided into control group， model group， dexamethasone group （positive control， 2 mg/kg）， CB low-dose and high-dose groups （25， 50 mg/kg）. Each group was given relevant medicine intraperitoneally， once a day， for 3 consecutive days. Twenty-four hours after the last administration， those groups were given lipopolysaccharide （10 mg/kg） intraperitoneally to establish sepsis-induced ALI model （finally， 8 mice per group were included in the experiment）， except for control group. Twenty-four hours after medication， blood routine indicators （total white blood cell count， neutrophils count， lymphocytes count）， lung function indicators （total lung resistance， pulmonary outflow resistance， lung dynamic compliance， peak expiratory flow rate， and maximum ventilation volume）， dry wet ratio of lung tissue were measured in each group. The lung tissue level of myeloperoxidase （MPO）， and the serum levels of tumor necrosis factor-α （TNF-α）， interleukin 1β （IL-1β）， IL-6， superoxide dismutase （SOD） and malondialdehyde （MDA） were all detected. The pathological changes of lung tissue were observed； immunohistochemistry was used to detect the positive expression of phosphorylation signal transducer and activator of transcription 3 （p-STAT3） in the lung tissue. Western blot assay was used to detect the expressions of proteins related to IL-6/JAK2/ STAT3 signaling pathway in the lung tissue. RESULTS Compared with control group， total pulmonary resistance， pulmonary flow resistance， dry wet ratio of lung tissue， the total white blood cell count， neutrophils count， lymphocytes count of whole blood， the lung tissue level of MPO and serum levels of MDA， IL-6， IL-1β and TNF-α， the p-STAT3 protein optical density value， the protein expressions of IL-6 and IL-6 receptor， and the phosphorylation levels of JAK2 and STAT3 protein were increased significantly in the model group （P＜0.01）， while lung dynamic compliance， peak expiratory flow rate， maximum ventilation volume and serum level of SOD were decreased significantly （P＜0.05 or P＜0.01）. Pulmonary tissue showed alveolar collapse and infiltration of inflammatory cells. Compared with the model group， the above indexes of mice were reversed significantly in dexamethasone group and CB groups （P＜0.05 or P＜0.01）， the pathological damage of lung tissue was reduced. CONCLUSIONS CB can prevent sepsis-induced ALI by inhibiting the activity of Δ 基金项目遵义市科技计划项目（No.202252） IL-6/JAK2/STAT3 signaling pathway and relieving ＊第一作者主治医师。研究方向：重症医学。E-mail：fjuanxui@ 163.com inflammatory reactions. # 通信作者 主任医师。研究方向：儿童呼吸系统疾病诊断与治

Objective To explore the effect and mechanism of hesperidin in treating the lung injury in the mouse model of respiratory syncytial virus (RSV)-induced bronchiolitis. Methods A mouse model of RSV-induced bronchiolitis was established,and 60 BALB/c mice were assigned into a control group,a model group,a low-dose hesperidin (18 mg/kg) group,a high-dose hesperidin (36 mg/kg) group,and a high-dose hesperidin (36 mg/kg)+Jagged1(1 mg/kg) group by random number table method,with 12 mice in each group. Corresponding doses of drugs were administrated for intervention,and the control group and model group were administrated with the same amount of saline.The bronchoalveolar lavage fluid (BALF) samples were collected and alveolar macrophages were isolated.ELISA was employed to detect the levels of interleukin (IL)-4,IL-6,tumor necrosis factor-α (TNF-α),and IL-10 in BALF,and flow cytometry to detect the M1/M2 polarization of macrophages.qRT-PCR and Western blotting were respectively conducted to detect the mRNA and protein levels of inducible nitric oxide synthase (iNOS),arginase 1 (Arg-1),Jagged1,and Notch1 in the lung tissue. Results Compared with the control group,the modeling of RSV-induced bronchiolitis elevated the IL-4,IL-6,and TNF-α levels,increased the proportion of M1-type macrophages and the lung inflammation and mucus secretion scores,and up-regulated the mRNA and protein levels of iNOS,Jagged1,and Notch1 in BALF (all P<0.001).Meanwhile,the modeling lowered the IL-10 level,decreased the proportion of M2-type macrophages,and down-regulated the mRNA and protein levels of Arg-1 (all P<0.001).Compared with the model group,low- and high-dose hesperidin lowered the IL-4,IL-6,TNF-α levels,decreased the proportion of M1-type macrophages and the lung inflammation and mucus secretion scores,and down-regulated the mRNA and protein levels of iNOS,Jagged1,and Notch1 in BALF (all P<0.05).Moreover,hesperidin elevated the IL-10 level,increased the proportion of M2-type macrophages,and up-regulated the mRNA and protein levels of Arg-1 (all P<0.001).Using recombinant Jagged1 protein to activate Notch1 signaling pathway can significantly attenuate the promotion of high-dose hesperidin on M2 macrophage polarization and amelioration of lung inflammation damage (all P<0.01). Conclusion Hesperidin may alleviate the lung inflammation damage in mice with RSV-induced bronchiolitis by inhibiting the Jagged1/Notch1 signaling pathway and promoting the M2-type polarization of macrophages.
Animals ; Mice ; Bronchiolitis/metabolism* ; Hesperidin/metabolism* ; Interleukin-10/pharmacology* ; Interleukin-4/pharmacology* ; Interleukin-6/metabolism* ; Jagged-1 Protein/pharmacology* ; Lung Injury/metabolism* ; Macrophages ; Mice, Inbred BALB C ; RNA, Messenger/metabolism* ; Tumor Necrosis Factor-alpha/metabolism*