Objective·To investigate the regulatory effects and underlying mechanisms of mouse mandibular osteoblasts on B cell differentiation and development.Methods·Single-cell suspensions from mouse mandibular bone were prepared using an optimized enzymatic digestion method and induced to differentiate into osteoblasts in vitro.Osteogenic potential was validated by real-time quantitative PCR(RT-qPCR),alkaline phosphatase(ALP)staining,and alizarin red S(ARS)staining.The spatial localization relationship between osteoblasts and B cells in mandibular tissues was examined via immunofluorescence staining.High-purity hematopoietic progenitor cells were isolated using fluorescence-activated cell sorting.A Transwell co-culture system was established to assess the regulatory effects of different osteoblast concentrations(5×104,2.5×105,and 5×105 cells/well)on B cell differentiation(5×104 cells/well).Flow cytometry and RT-qPCR were employed to evaluate B cell viability and differentiation.Additionally,RT-qPCR was used to analyze the expression of osteoblast-secreted factors associated with B cell development during osteogenic differentiation.Results·Mandibular osteoblasts exhibited robust osteogenic potential,as confirmed by ALP/ARS staining and high expression of osteogenic markers(Runx2,Osx,Ocn,and Alp)via RT-qPCR.Immunofluorescence revealed close spatial proximity between osteoblasts and B cells in mandibular tissues.In the co-culture system,osteoblasts promoted B cell differentiation in a concentration-dependent manner.RT-qPCR and immunofluorescence demonstrated that osteoblasts significantly upregulated key genes involved in B cell development(Ebf1,Rag1,Il7r,and Pax5;all P<0.001).Furthermore,osteoblast-derived factors(Il7,Baff,and Flt3l)were markedly elevated during osteogenic differentiation(all P<0.05).Conclusion·Mandibular osteoblasts enhance B cell differentiation and development in a concentration-dependent manner,likely through secreting growth factors that upregulate critical B cell differentiation genes.

Objective·To investigate the regulatory effects and underlying mechanisms of mouse mandibular osteoblasts on B cell differentiation and development.Methods·Single-cell suspensions from mouse mandibular bone were prepared using an optimized enzymatic digestion method and induced to differentiate into osteoblasts in vitro.Osteogenic potential was validated by real-time quantitative PCR(RT-qPCR),alkaline phosphatase(ALP)staining,and alizarin red S(ARS)staining.The spatial localization relationship between osteoblasts and B cells in mandibular tissues was examined via immunofluorescence staining.High-purity hematopoietic progenitor cells were isolated using fluorescence-activated cell sorting.A Transwell co-culture system was established to assess the regulatory effects of different osteoblast concentrations(5×104,2.5×105,and 5×105 cells/well)on B cell differentiation(5×104 cells/well).Flow cytometry and RT-qPCR were employed to evaluate B cell viability and differentiation.Additionally,RT-qPCR was used to analyze the expression of osteoblast-secreted factors associated with B cell development during osteogenic differentiation.Results·Mandibular osteoblasts exhibited robust osteogenic potential,as confirmed by ALP/ARS staining and high expression of osteogenic markers(Runx2,Osx,Ocn,and Alp)via RT-qPCR.Immunofluorescence revealed close spatial proximity between osteoblasts and B cells in mandibular tissues.In the co-culture system,osteoblasts promoted B cell differentiation in a concentration-dependent manner.RT-qPCR and immunofluorescence demonstrated that osteoblasts significantly upregulated key genes involved in B cell development(Ebf1,Rag1,Il7r,and Pax5;all P<0.001).Furthermore,osteoblast-derived factors(Il7,Baff,and Flt3l)were markedly elevated during osteogenic differentiation(all P<0.05).Conclusion·Mandibular osteoblasts enhance B cell differentiation and development in a concentration-dependent manner,likely through secreting growth factors that upregulate critical B cell differentiation genes.

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 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.