Small RNAs (sRNAs), the key components of RNA interference (RNAi) or RNA silencing, can mediate cell-autonomous gene silencing and function as signaling molecules across species. Microbe-induced gene silencing (MIGS), which is based on interspecies RNAi, is an effective approach for controlling fungal diseases in crops. The enolase gene VdEno is essential for the growth and development of the fungal pathogen Verticillium dahliae, which causes cotton Verticillium wilt. In this study, we engineered Trichoderma harzianum (Th) to express the double-stranded RNA (dsRNA) targeting VdEno. The engineered strain Th-VdEnoi successfully generated VdEno-specific small interfering RNA (siVdEno). We further confirmed that Th-VdEnoi effectively induced VdEno silencing at the translational level. The results of crop protection assays revealed that the cotton plants co-inoculated with V. dahliae (strain V592) and Th-VdEnoi presented significantly reduced disease severity and lower fungal biomass in their roots than the control plants inoculated with V. dahliae alone or with V. dahliae and Th-GFPi (a control strain expressing GFP-targeting dsRNA). Collectively, our findings demonstrate that VdEno is an effective target for controlling cotton Verticillium wilt and confirm that MIGS is a promising strategy for managing soil-borne fungal pathogens in crops. MIGS provides strong technical support for reducing the application of conventional chemical pesticides, developing eco-friendly biopesticides, and facilitating the sustainable development of agriculture.
Gossypium/microbiology* ; Plant Diseases/prevention & control* ; Gene Silencing ; Ascomycota/genetics* ; RNA Interference ; RNA, Double-Stranded/genetics* ; Hypocreales/genetics* ; RNA, Small Interfering/genetics* ; Verticillium/genetics* ; Fungal Proteins/genetics*

The comparison between traditional Chinese medicine Jinzhen oral liquid (JZOL) and Western medicine in treating children with acute bronchitis (AB) showed encouraging outcomes. This trial evaluated the efficacy and safety of the JZOL for improving cough and expectoration in children with AB. 480 children were randomly assigned to take JZOL or ambroxol hydrochloride and clenbuterol hydrochloride oral solution for 7 days. The primary outcome was time-to-cough resolution. The median time-to-cough resolution in both groups was 5.0 days and the antitussive onset median time was only 1 day. This randomized controlled trial showed that JZOL was not inferior to cough suppressant and phlegm resolving western medicine in treating cough and sputum and could comprehensively treat respiratory and systemic discomfort symptoms. Combined with clinical trials, the mechanism of JZOL against AB was uncovered by network target analysis, it was found that the pathways in TRP channels like IL-1β/IL1R/TRPV1/TRPA1, NGF/TrkA/TRPV1/TRPA1, and PGE2/EP/PKA/TRPV1/TRPA1 might play important roles. Animal experiments further confirmed that inflammation and the immune regulatory effect of JZOL in the treatment of AB were of vital importance and TRP channels were the key mechanism of action.

Objective To discuss the correlation between dynamic changes in pulmonary function and chronic cough in 5-to-14-year-old children with Mycoplasma pneumoniae pneumonia(MPP)after acute period(about 2 weeks after admission). Methods One hundred and fifty - six hospitalized children diagnosed with MPP from Lianyungang Maternal and Child Health Care Hospital Affiliated to Yangzhou University from February 2014 to May 2017 were selected. According to the results of routine ventilatory pulmonary function before discharge(10-14 days in hospital;acute phase group),there were 50 patients with normal pulmonary function and 106 patients with abnormal pulmonary function. All patients continued to take oral azithromycin for 2 to 3 courses after discharge,and the indexes of lung function were dynamically tracked in 1 month(1-month group)and 2 months(2-month group)in the group of abnormal lung function after leaving the hospital,and the incidences and causes of chronic cough were followed up. Results (1)There were 106 cases with abnormal pulmonary function in 156 cases with MPP in acute phase group,and the rate of incidence was 67. 95%. Twenty-seven(29. 35%)out of 97 children were still abnormal in pulmonary functional testing 1 month after leaving hospital( 1-month group). Among the 27 cases,about 18. 52%(5/27 cases) of them still did not return to normal 2 months after discharge( 2-month group ). There were significant differences in the occurrence of abnormal pulmonary function among 3 groups mentioned above(χ2 ＝162. 64,P<0. 001).(2)Ratios of measured values and predicted ones of forced vital capacity( FVC ),forced expiratory volume in one second (FEV1 ),peak expiratory flow(PEF)and maximum mid-expiratory flow(MMEF 25% -75%)in the lung function of 2-month group after the acute phase of MPP were significantly higher than those of 1-month group and acute phase group,and the values of 1 -month group were better than those in acute phase one,which were statistically different among 3 groups(P<0. 01).(3)The rate of occurrence of chronic cough in normal lung function group was about 18. 00%(9/50 cases),and in abnormal pulmonary function group,it was about 70. 75%(75/106 cases). There was a significant difference between them(χ2 ＝35. 96,P<0. 05). Abnormal pulmonary functions were the influencing factors of chronic cough(r＝0. 55,P<0. 01).(4)There were 5 cases with upper airway cough syndrome(UACS)and 4 cases with post-infection cough(PIC)found in the normal lung function group. By contrast,75 cases suffered from chronic cough in abnormal pulmonary functions group,of which 36 cases with cough variant asthma( CVA),24 cases with UACS,8 cases with comorbidity of CVA and UACS and 7 cases with PIC. Conclusions Abnormal lung function after acute period of MPP may last 4 to 8 weeks,or probably even longer. In normal lung function group,UACS is the most common cause,then followed by PIC. On the contrary,the main cause of chronic cough in abnormal pulmonary function group is CVA,followed by UACS. Chronic cough is related to abnormal lung function after acute phase of MPP.

Asthma is a common chronic pulmonary disease in children,and most of the children with asthma can be well controlled after standardized treatment. But there are some children with difficult-to-treat asthma in cli-nical,although the high dose of glucocorticoid atomization inhalation treatment,the clinical symptoms are still not well controlled. These children with difficult-to-treat asthma not only bring huge economic pressure to families,but also have irreversible lung function impairment and their quality of life as the disease progresses. Therefore,pediatricians need to identify the difficult-to-treat asthma,understand its etiology,and give appropriate treatment to improve the prognosis of these patients.

Objective To investigate the roles of fiberoptic bronchoscopy in diagnosis and treatment for infants with refractory and persistent wheezing. Methods From Jun. 2012 to Dec. 2013, 52 hospitalized children with age between four 4 months and 1 year old were recruited for ifberoptic bronchoscopy, who had been wheezing for at least four weeks and treated ineffectively with conventional anti-inlfammatory agents:budesonide and compound ipratropium bromide solution. Then, the pathogenesis of refractory and persistent wheezing was summarized based on clinical features, detection of CT imaging of three-dimensional airway reconstruction and cardiac CT, results of bronchoscopy inspection, and bronchoalveolar lavage lfuid culture. Results Among the 52 cases, 40 were with ground glass-like changes (76.92%) in pulmonary spiral CT testing, 4 with mosaic perfusion syndrome (7.69%), 8 with segmental pulmonary consolidation (15.38%), 8 with obstructive pulmonary emphysema (15.38%), and 1 with left primary bronchial foreign body. In addition, through bronchofibroscopy, there were 52 cases with imlfammation (100%),3 with tracheal stenosis (5.77%), 3 with left and/or right main bronchus stenosis of the external pressure, 18 with bronchomalacia(34.62%), 2 cases with foreign body (3.84%), one in trachea (1.92%), the other in left main bronchus (1.92%), 10 with bronchial mucus plug (19.23%), and 8 (15.38%) with congenital airway malformations (including 3 at tracheal bronchus, 1 at left upper lobe bronchial stenosis and 1 at bronchial Bridge). The culture of bronchoalveolar lavage lfuid were conducted for all patients. The positive rate of bronchoalveolar lavage lfuid was 9.62%(5/52 cases), including 2 cases with tip Escherichia coli, 2 with Haemophilus inlfuenzae, and 1 with Acinetobacter baumannii. Conclusions First, infection is the primary cause of refractory and persistent wheezing, which is persistent in airway resulted from multi-drug resistant bacteriua. Second, refractory and persistent wheezing is often caused by multi-factors including infection, congenital airway malformations, the endogenous and exogenous foreign body, cardiovascular malformation, etc. These factors often lead to dififcult wheezing control. The last, the diagnosis rate of the refractory and persistent wheezing can be improved by combination of ifberoptic bronchoscopy and lung spiral CT.