肺力咳合剂联合雾化吸入布地奈德混悬液治疗小儿急性支气管炎的疗效及对血清急性时相蛋白和免疫球蛋白的影响

摘要 目的：观察肺力咳合剂联合雾化吸入布地奈德混悬液治疗急性支气管炎患儿的疗效及对血清急性时相蛋白和免疫球蛋白的影响。方法：选取2016年1月到2021年12月期间在西安医学院第二附属医院进行治疗的120例急性支气管炎患儿,按照随机数字表法分为对照组（接受雾化吸入布地奈德混悬液治疗,n=60）和研究组（肺力咳合剂联合雾化吸入布地奈德混悬液治疗,n=60）,对比两组疗效、用药不良反应情况,观察两组患儿临床症状缓解时间,观察两组患儿血清急性时相蛋白和免疫球蛋白的变化情况。结果：研究组的临床总有效率相较于对照组,可进一步升高,组间对比差异有统计学意义（P<0.05）。研究组的呼吸困难、咳嗽、肺部喘鸣、憋喘症状消失时间均短于对照组（P<0.05）。治疗后,两组触珠蛋白（HP）、C反应蛋白（CRP）、a1酸性糖蛋白（a1-AAG）和铜蓝蛋白（CER）水平均下降,且研究组低于对照组（P<0.05）。治疗后,两组免疫球蛋白A（IgA）、免疫球蛋白M（IgM）、免疫球蛋白G（IgG）均升高,且研究组较对照组高（P<0.05）。两组不良反应发生率对比无差异（P>0.05）。结论：肺力咳合剂联合雾化吸入布地奈德混悬液治疗急性支气管炎患儿,可有效改善体液免疫,降低患儿血清急性时相蛋白,疗效显著。     

三拗片联合布地奈德混悬液雾化吸入对急性支气管炎患儿炎症细胞因子和T淋巴细胞亚群的影响

摘要 目的：观察布地奈德混悬液雾化吸入、三拗片联合治疗对急性支气管炎患儿T淋巴细胞亚群、炎症细胞因子的影响。方法：选取2021年1月到2023年1月期间合肥市第二人民医院收治的急性支气管炎患儿78例,按照双色球法将患儿分为对照组和研究组,各为39例。对照组患儿接受布地奈德混悬液雾化吸入,研究组患儿接受三拗片联合布地奈德混悬液雾化吸入。对比两组疗效、炎症因子水平、临床指标、T淋巴细胞亚群和用药安全性。结果：研究组的临床总有效率高于对照组（P<0.05）。研究组的喘息缓解时间、咳嗽缓解时间、体温正常缓解时间、肺部啰音缓解时间短于对照组（P<0.05）。治疗7 d后,两组降钙素原（PCT）、白细胞介素-8（IL-8）、白细胞介素-6（IL-6）下降,且研究组低于对照组（P<0.05）。治疗7 d后,两组CD8⁺下降,且研究组低于对照组（P<0.05）;两组CD4⁺、CD4⁺/CD8⁺升高,且研究组高于对照组（P<0.05）。两组患儿不良反应对比未见统计学差异（P>0.05）。结论：急性支气管炎患儿采用布地奈德混悬液雾化吸入联合三拗片治疗,有助于缩短临床症状缓解时间,减轻炎症细胞因子水平,调节T淋巴细胞亚群。     

Identification of one peptide which inhibited infectivity of avian infectious bronchitis virus in vitro

Purified avian infectious bronchitis virus (IBV) was used to screen a random phage display peptide library. After the fourth panning, 10 positive phages were sequenced and characterized. The phages specifically inhibited IBV infectivity in HeLa cells and blocked IBV haemagglutination. One linear peptide “GSH HRH VHS PFV” from the positive phages with the highest neutralization titer was synthesized and this peptide inhibited IBV infection in HeLa as well. The results may contribute to development of antiviral therapeutics for IBV and studying the determinants for viral and cell interac-tion.     

鸡传染性支气管炎病毒变异株（793／B）核蛋白基因的克隆与序列分析

根据GenBank已经发表的传染性支气管炎病毒(IBV)N全基因组序列设计引物,对IBV793/B分离毒株N基因进行克隆与序列分析。结果表明,IBV793/B的N基因由1229bp组成,与GenBank已发表的11株IBV的N基因相比较,IBV793/B的N基因共有88处点突变,在第991位发生了一个核苷酸的缺失。N基因的核苷酸同源性为86.9%～91.4%,氨基酸同源性为75.8%～77.5%。表明IBV93/B的N基因存在着较大的变异性。     

Anti-oxidative effect of triterpene acids of Eriobotrya japonica (Thunb.) Lindl. leaf in chronic bronchitis rats

The study was to evaluate the effect of triterpene acids of Eriobotrya japonica (Thunb.) Lindl. leaf (TAL) on expression of antioxidative mediators by alveolar macrophages (AM) in rats with chronic bronchitis (CB), CB was induced by endotracheal instillation of lipopolysaccharedes (LPS) followed by Bacillus Calmette-Guérin (BCG) injection through caudal vein 1 week later. Treatment groups received TAL at there different doses (50, 150, or 450 mg/kg daily, intragastrically (i.g.)) or dexamethasone (1.2 mg/kg daily i.g.) for 2 weeks, 7 days after LPS injection. AM were then isolated and incubated. Superoxide dismutase (SOD) and methylene dianiline (MDA) levels in AM were measured by commercial kits; meanwhile, heme oxygenase-1 (HO-1) expression and its mRNA expression in AM were detected by immunocytochemistry and RT-PCR, respectively. HO-1 activity of the lung was also detected by a specific biochemistry reaction. The levels of MDA and HO-1 expressed by cultured AM and the HO-1 activity in the lung of the TAL groups were significantly lower than those from the CB group without treatment (p < 0.01 and p < 0.05, respectively), while the SOD levels were increased in a dose-dependent manner by TAL treatment. These results suggest that TAL inhibits HO-1 expression and MDA production and up-regulates SOD expression in AM from CB rats, which might be one of molecular mechanisms of its anti-inflammatory effects in CB rats.     

几种鸡传染性支气管炎病毒活疫苗对中国流行毒株CK/CH/LDL/97I的保护作用评价

选择我国应用的五株鸡传染性支气管炎活疫苗毒株(JAAS、IBN、Jlin、J9和H120)和当地流行毒株(CK/CH/LDL/97 Ⅰ)作为研究对象,对其S1基因进行序列比对分析,结果表明疫苗株与流行毒株的核昔酸序列及推导的氨基酸序列同源性分别不超过76.4%和78.7%.S1基因的核苷酸系统发育树显示,疫苗株与流行毒株分属不同进化群,亲缘关系较远,属于不同的基因型.用这五株活疫苗进行针对强毒株CK/CH/LDL/97Ⅰ株的免疫保护实验,可见临床发病率为30%～100%;攻毒5d后每组随机扑杀10只鸡,采集器官,应用RT-PCR法检测病毒,气管样品病毒检出率为50%～90%,肾脏样品病毒检出率为10%～30%.由此可见:我国目前使用的主要活疫苗对异种IBV分离株的感染不能提供完全的保护作用.     

α-Galactosidase from Alfalfa Seeds (Medicago sativa L.)

An α-galactosidase from alfalfa seeds was purified 140-fold by ammonium sulfate fractionation, and column chromatography on Sephadex G-100, DEAE- and CM-Sephadex. Polyacrylamide-gel electrophoresis of the purified enzyme showed a single protein band. The molecular weight was estimated to be approximately 57,000 by gel-filtration. The purified enzyme hydrolyzed p-nitrophenyl α-d-galactoside more rapidly than raffinose. The maximal enzyme activities were obtained at pH 4.0 and 5.5 for p-nitrophenyl α-d-galactoside and at 4.5 for raffinose. The enzyme was shown to be inhibited by Hg²⁺ and Ag⁺ ions, and d-galactose.     

Development of a multi-epitope antigen of S protein-based ELISA for antibodies detection against infectious bronchitis virus

An indirect enzyme-linked immunosorbent assay (ELISA) method based on a novel multi-epitope antigen of S protein (SE) was developed for antibodies detection against infectious bronchitis virus (IBV). The multi-epitope antigen SE protein was designed by arranging three S gene fragments (166–247 aa, S1 gene; 501–515 aa, S1 gene; 8–30 aa, S2 gene) in tandem. It was identified to be approximately 32 kDa as a His-tagged fusion protein and can bind IBV positive serum by western blot analysis. The conditions of the SE-ELISA method were optimized. The optimal concentration of the coating antigen SE was 3.689 μg/mL and the dilution of the primary antibodies was identified as 1:1000 using a checkerboard titration. The cut-off OD₄₅₀ value was established at 0.332. The relative sensitivity and specificity between the SE-ELISA and IDEXX ELISA kit were 92.38 and 89.83%, respectively, with an accuracy of 91.46%. This assay is sensitive and specific for detection of antibodies against IBV.     

Visualizing the autophagy pathway in avian cells and its application to studying infectious bronchitis virus

Autophagy is a highly conserved cellular response to starvation that leads to the degradation of organelles and long-lived proteins in lysosomes and is important for cellular homeostasis, tissue development and as a defense against aggregated proteins, damaged organelles and infectious agents. Although autophagy has been studied in many animal species, reagents to study autophagy in avian systems are lacking. Microtubule-associated protein 1 light chain 3 (MAP1LC3/LC3) is an important marker for autophagy and is used to follow autophagosome formation. Here we report the cloning of avian LC3 paralogs A, B and C from the domestic chicken, Gallus gallus domesticus, and the production of replication-deficient, recombinant adenovirus vectors expressing these avian LC3s tagged with EGFP and FLAG-mCherry. An additional recombinant adenovirus expressing EGFP-tagged LC3B containing a G120A mutation was also generated. These vectors can be used as tools to visualize autophagosome formation and fusion with endosomes/lysosomes in avian cells and provide a valuable resource for studying autophagy in avian cells. We have used them to study autophagy during replication of infectious bronchitis virus (IBV). IBV induced autophagic signaling in mammalian Vero cells but not primary avian chick kidney cells or the avian DF1 cell line. Furthermore, induction or inhibition of autophagy did not affect IBV replication, suggesting that classical autophagy may not be important for virus replication. However, expression of IBV nonstructural protein 6 alone did induce autophagic signaling in avian cells, as seen previously in mammalian cells. This may suggest that IBV can inhibit or control autophagy in avian cells, although IBV did not appear to inhibit autophagy induced by starvation or rapamycin treatment.     

Genetic susceptibility for chronic bronchitis in chronic obstructive pulmonary disease

Background

Chronic bronchitis (CB) is one of the classic phenotypes of COPD. The aims of our study were to investigate genetic variants associated with COPD subjects with CB relative to smokers with normal spirometry, and to assess for genetic differences between subjects with CB and without CB within the COPD population.

Methods

We analyzed data from current and former smokers from three cohorts: the COPDGene Study; GenKOLS (Bergen, Norway); and the Evaluation of COPD Longitudinally to Identify Predictive Surrogate Endpoints (ECLIPSE). CB was defined as having a cough productive of phlegm on most days for at least 3 consecutive months per year for at least 2 consecutive years. CB COPD cases were defined as having both CB and at least moderate COPD based on spirometry. Our primary analysis used smokers with normal spirometry as controls; secondary analysis was performed using COPD subjects without CB as controls. Genotyping was performed on Illumina platforms; results were summarized using fixed-effect meta-analysis.

Results

For CB COPD relative to smoking controls, we identified a new genome-wide significant locus on chromosome 11p15.5 (rs34391416, OR = 1.93, P = 4.99 × 10^-8) as well as significant associations of known COPD SNPs within FAM13A. In addition, a GWAS of CB relative to those without CB within COPD subjects showed suggestive evidence for association on 1q23.3 (rs114931935, OR = 1.88, P = 4.99 × 10^-7).

Conclusions

We found genome-wide significant associations with CB COPD on 4q22.1 (FAM13A) and 11p15.5 (EFCAB4A, CHID1 and AP2A2), and a locus associated with CB within COPD subjects on 1q23.3 (RPL31P11 and ATF6). This study provides further evidence that genetic variants may contribute to phenotypic heterogeneity of COPD.

Trial registration

ClinicalTrials.gov {"type":"clinical-trial","attrs":{"text":"NCT00608764","term_id":"NCT00608764"}}NCT00608764, {"type":"clinical-trial","attrs":{"text":"NCT00292552","term_id":"NCT00292552"}}NCT00292552

Electronic supplementary material

The online version of this article (doi:10.1186/s12931-014-0113-2) contains supplementary material, which is available to authorized users.