Objective To observe the eotaxin expression of rat airway smooth muscle cells ( ASMCs) induced by serum from asthmatic rats, and explore the possible mechanism. Methods ASMCs isolated fromrat tracheas were cultured in vivo. Then they were treated with serum from asthmatic rats, or treated with serum and dexamethasone simultaneously. The level of eotaxin protein in supernatant and eotaxin mRNA in ASMCs were measured by ELISA and reverse transcription-polymerase chain reaction. The expression of cAMP in ASMCs was examined by radioimmunoassay. Results After the treatment with sensitized serum, the eotaxin level in supernatant and mRNA expression in ASMCs were significantly higher [ ( 107. 09 ±7. 12) ng/L vs. ( 0. 63 ±0. 56) ng/L, P lt; 0. 05; 1. 39 ±0. 04 vs. 0. 05 ±0. 01, P lt;0. 05] , and the level of cAMP in ASMCs was significantly lower compared with the control group [ ( 17. 58 ±3. 62) ng/L vs. ( 32. 39 ±3. 36) ng/L, P lt; 0. 05] . After intervened by the sensitized serum and dexamethasone simultaneously, the protein and mRNA expressions of eotaxin were lower compared with those intervened by sensitized serumalone [ ( 64. 18 ±4. 04) ng/L and 0. 77 ±0. 19] . The level of eotaxin in supernatant was negatively correlated with cAMP level in ASMCs ( r = - 0. 788, P lt; 0. 01) . Conclusions There is anautocrine function in ASMCs as inflammatory cells after stimulation with sensitized serum. Eotaxin may play an important roll in the pathogenesis of asthma via a cAMP-dependent pathway.
Objective To investigate the expression of stromal cell derived factor-1 ( SDF-1) and the effects of budesonide suspension for inhalation ( Pulmicort Respules) in mice with asthma. Methods Thirty Kunming female mice were randomly divided into three groups, ie. a control group, an asthma group, and a pulmicort treatment group. The asthma group and the pulmicort treatment group were sensitized with ovalbumin ( OVA) by a combination of intraperitoneal injection and repeated OVA intranasal challenges to establish mouse asthma model. The pulmicort treatment group received 100μL pulmicort by intranasal administration before OVA challenge. The immunohistochemistry was used to estimate the expression of SDF-1 in lung tissues. HE staining and Wright-Giemsa staining method were used to assess inflammatory infiltration in the airway and bronchoalveolar lavage fluid ( BALF) respectively. Results The expression of SDF-1 in the asthma group increased significantly compared with the control group ( 0.48 ±0.03 vs. 0.21 ± 0.02, Plt;0.05) , and significantly decreased after the intervention with pulmicort ( 0.29 ±0.01 vs. 0.48 ± 0.03, Plt; 0.05 ) . Compared with control group, the infiltration of inflammatory cells in airway was significantly enhanced in the asthma group, and attenuated in the pulmicort treatment group. The total number of inflammatory cells and eosinophil, lymphocyte, neutrophil counts in BALF increased significantly in the asthma group compared with the control group, and decreased significantly after pulmicort intervention. Conclusion SDF-1 may play an important role in the recruitment of inflammatory cells in asthmatic airway and pulmicort may relieve airway inflammation by decreasing the expression of SDF-1.
ObjectiveTo observe repairing process of trachea epithelium cells in chlorine-induced airway epithelial injury.MethodsTwelve mice were exposed to chlorine gas and prepared the mice model of airway damage. Three mice were executed respectively on 2nd, 4th, 7th, 10th day after exposure to chlorine gas, and tracheal tissues were collected. In addition 3 normal mice served as control. Airway repair and cell proliferation were detected by EdU labeling method. The basal cell markers keratin 5 (K5), keratin 14 (K14) were adopted as the tracheal epithelial markers for locating the position of the proliferation of repairing cells. Morphological analysis was adopted to measure the proliferation rate as well as the recovery of the false stratified epithelium.ResultsIn the control group, cell proliferation rate was very low, all basal cells expressed K5, and most basal cells did not express K14. Most of epithelial cells shed from the trachea epithelium after exposure to chlorine gas. 2-4 days after chlorine exposure, K5 and K14 expression basal cells increased, K14 expression cells increased greatly. In the peak period of cell proliferation, only a small number of ciliated cells appeared in the repairing trachea area. Epithelial cells repaired fast and widely at the bottom of the trachea.ConclusionThe trachea residual basal cells play roles of progenitor cells and repair the airway epithelium after chlorine damage in mice.
ObjectiveTo study immunodepression effect of bone marrow-derived mesenchymal stem cell (BMSC) on acute asthmatic airway inflammation by galectin-1 (gal-1) in vivo.MethodsEighty-five female BALB/c mice were equally randomized into normal control group, asthmatic group, BMSC treatment group, gal-1 treatment group and BMSC and gal-1 inhibitor group. Ovalbumin (OVA) was used to establish acute asthmatic model. Total cell number and differential cell analysis in each group in bronchoalveolar lavage fluid (BALF) were determined. Furthermore, hematoxylin-eosin and periodic-acid Schiff staining was used to compare airway inflammation among five groups. Measurement of cytokines, including interleukin (IL) -4, IL-5 and gal-1 in BALF and OVA specific IgE (OVA-IgE) in serum were evaluated by enzyme linked immunosorbent assay. Moreover, dendritic cell (DC) in lung tissue was sorted by immunomagnetic beads and its MAPK signal pathway was analyzed by western blotting among five groups.ResultsAccumulation of inflammation cells, particularly eosinophils around airway and in BALF was evident in asthmatic mouse model, meanwhile hyperplasia of Goblet cell was also obvious in asthmatic group. BMSC engraftment or gal-1 infusion significantly reduced airway inflammation and hyperplasia of Goblet cell and the number of inflammation cells in BALF, especially eosinophils attenuated dramatically. However, there was no effect on airway inflammation and hyperplasia of Goblet Cell by simultaneous infusion BMSC engraftment and gal-1 inhibitor. Compared to normal control group, the level of IL-4, IL-5 in BALF and OVA-IgE in serum was increased remarkably in asthmatic group, but the level of gal-1 reduced obviously. Moreover, infusion of BMSC or gal-1 could mitigate the level of IL-4, IL-5 in BALF and OVA-IgE in serum and increase the level of gal-1 in asthmatic mouse. However, infusion with both BMSC and gal-1 inhibitor exerted no effect on cytokine and OVA-IgE in asthmatic mouse. DC was sorted by immunomagnetic beads and western blotting was used to detect the expression of MAPK signal pathway among five groups. The expression of ERK phosphorylation in asthmatic group was much lower than that in normal control group. On the contrary, the expression of p38 phosphorylation was much higher than that in normal control group. BMSC engraftment or gal-1 infusion significantly activated the ERK pathway and inhibited the p38 MARP pathway on asthmatic mouse DC. Nevertheless, the expression of ERK phosphorylation and p38 phosphorylation for group with BMSC and gal-1 inhibitor infusion was between the level of asthmatic group and normal control group.ConclusionsBMSC infusion alleviates airway inflammation in asthmatic mouse, especially weakens eosinophils infiltration, and the underlying mechanism might be protective effect of gal-1 secreted by BMSC which plays a role in lung tissue DC and regulates the DC expression of MAPK signal pathway.
Objective To explore the diagnosis and treatment of airway involvement in relapsing polychondritis. Methods The clinical data of two patients with relapsing polychondritis with airway involvement were reported and the relative literatures were reviewed. Results The two patients were both old males, with clinical manifestations of cough, dyspnea, and fever. They were misdiagnosed in a other hospital. The pulmonary function tests showed obstructive ventilatory impairemnt. On inspiratory CT, tracheal / tracheobronchial wall thickening and airway stenosis, with or without tracheal cartilage calcification were common findings. The tracheal cartilages thickeness and membranous wall were normal. On expiratory CT scans, functional abnormalities were identified such as tracheobronchomalacia. The patients were relieved by medication of corticosteroids or with immunodepressant. Conclusions The relapsing polychondritis with airway involvement is easy to be misdiagnosed. Chest CT examination is a valuable method for diagnosis of relapsing polychondritis. Corticosteroids and immunodepressant can improve the outcome.
Objective To investigate the modulating roles of Clara cell secretory 16 kD protein ( CC-16) , transcription factor T-bet, and GATA-3 in airway inflammation of patients with asthma. Methods 25 patients with acute exacerbation of asthma were enrolled as an asthma group and 33 healthy volunteers were enrolled as control. The plasma levels of CC16, IFN-γ, and IL-4 were measured by enzyme-linked immunosorbent assay ( ELISA) . The mRNA expressions of T-bet and GATA-3 in the peripheral bloodmononuclear cells ( PBMCs) were measured by reverse transcription-polymerase chain reaction ( RT-PCR) .Results The levels of CC16 and IFN-γin the asthma group were lower than those in the control group [ ( 21. 96 ±7. 31 ) ng/mL vs. ( 64. 88 ±25. 27) ng/mL, ( 118. 73 ±22. 59) pg/mL vs. ( 145. 53 ±29. 50) pg/mL, both P lt;0. 01] . The IL-4 level in the asthma group was significantly higher than that in the control group [ ( 425. 22 ±4. 37) pg/mL vs. ( 69. 72 ±10. 15 ) pg/mL, P lt; 0. 01] . The T-bet mRNA expression and T-bet /GATA-3 ratio of PBMCs in the asthma group were significantly lower than those in the control group( both P lt; 0. 01) . The expression GATA-3 mRNA was significantly higher than that in the control group( P lt;0. 01) . The level of CC16 was positively correlated with T-bet mRNA expression and the ratio of T-bet /GATA-3 ( r =0. 792, 0. 761, respectively, P lt; 0. 01) . There was no correlation between CC16 and the GATA-3 mRNA expression ( P gt;0. 05) . Conclusions These results suggest that CC16 and T-bet play important protection roles in the pathogenesis of asthma. GATA-3, IFN-γ, and IL-4 also participate in the airway inflammation of asthma.
Objective To examine the effects of Pseudomonas aeruginosa(PA)quorum-sensing systems on airway mucus hypersecretion．Methods Sixty Sprague-Dawley rats were intubated with a silicone tube pre-coated with PAO1(wild-type PA strain)，PAO1-JP2(quorum-sensing-mutant strain)or saline in the bronchus．After 28 days，the mRNA and protein expression of MUC5AC in the rats’bronchial epithelia were detected by RT-PCR，alcian blue／periodic acid—Schif(AB／PAS)staining and enzyme linked immunosorbent assay(ELISA)．Results In the PAO1 group，bronchiolar epithelium goblet cells by AB／PAS staining was significantly more than those in the PAO1-JP2 and control groups(both Plt;0．05)．The expression level of MUC5AC mRNA in the PAO1 group was significantly higher than those in the PAO1-JP2 and control groups(both P lt;0．05)．The ELISA showed that the concentration of MUC5AC protein in bronchoalveolar lavage fluid(BALF)in the PAO1 group was much higher than that in the PAO1-JP2 group(P lt;0．05)．Conclusion PA quorum-sensing system plays an important role in airway mucus hypersecretion
Abstract: ObjectiveTo explore the surgical characteristics of primary tracheal tumors treatment and its prognosis. Methods [WTBZ]We retrospectively investigated the clinical records of 38 patients with primary tracheal tumors in both Xiangya Hospital of Central South University and Hunan Provincial Tumor Hospital from Febuary 1982 to August 2009. There were 24 males and 14 females aged from 7 to 65 years. There were 2 benign lesions, 13 adenoid cystic carcinomas, 11 squamous cell carcinomas, 5 mucoepidermoid carcinomas, 4 adenocarcinomas and 3 other cell types. One patient with adenocarcinoma underwent exploratory thoracotomy only; 33 patients underwent tracheal resection and airway reconstruction, and according to the tumor growth characteristics, the surgeon applied circumferential tracheal resection with endtoend anostomosis or wedge resection; One patient had papilloma resected under fiber bronchoscopy; and three patients with locally advanced lesions underwent radiotherapy without surgery. The overall survival rate was calculated by the KaplanMeier method. The logrank method was used for comparing survival rates among different groups, characterized by cell types or surgical procedures. Results [WTBZ]The patient with exploratory thoracotomy died 3 days after surgery from respiratory failure. The perioperative mortality was 2.94% (1/34), and all the remaining 33 patients recovered and were discharged from hospital. Minor complications happened to 12 patients (35.29%), including 6 patients with pulmonary infection, 4 with atelectasis, and 2 with hoarseness. The followup time ranged from 6 months to 15 years. The followup rate was 97.29% (36/37). The threeunresected patients died within 6 months after hospital discharge. The 1, 5, and 10year survival rate for resected patients was88% （95%CI 0.77 to 0.99), 47%(95%CI 0.29 to 0.66), and 41% (95%CI 0.21 to 0.61), respectively. The survival rate of adenoid cystic carcinoma or mucoepidermoid carcinoma was significantly higher than that of squamous cell carcinoma or other tissue types (χ2=17.581, P=0.001). There was no statistical difference (χ2=0.021, P=0.886) in 5 year survival rate between wedge resection group at 63%(95%CI 0.34 to 0.91) and the segmental resection group at 77%(95%CI 0.44 to 0.99). Conclusions [WTBZ]Surgical treatment is safe and beneficial for primary tracheal tumors, and the pathological type is a significant prognostic factor after complete resection.
Objective To investigate the application and long-termresults of epiglottic in reconstruction of the traumatic laryngotracheal stenosis.Methods From January 1988 to February 2002, 42 patients with traumatic laryngotracheal stenosis were treated, including 33 laryngeal stenosis and9 laryngotracheal stenosis. The following surgical treatment were performed: ① lowered epiglottic andbi-pedicled sternohyoid myofascial flap and ② lowered epiglottic and bipedicledsternohyoid myofascial flap and sternocleidomastoideus clavicle membrane flap. Results Thirty-seven patients(88.1%) were successfully decannulated 10 to 75 daysafter operation. Feeding tube lasted from 9 to 24 days, all the patients rehabilitated deglutition after surgery. The time of using stent was 9 to 19 days in 25cases.All patients were followed up 1 year to 3 years and 4 months. The function of larynx recovered completely in 37 decannulated patients and partially in 5cannulated patients. Conclusion Epiglottic- has the advantages of easy gain, high antiinfection and survival rate, and stable structure. A combination of epiglottic and the bipedicled sternohyoid myofascial flap plus sternocleidomastoideus clavicle membrane flap can repair large laryngeal and tracheal defects.
ObjectiveTo establish a method of air-liquid interface culture and ciliary beat frequency measurement of mouse tracheal-bronchial epithelial cells to simulate the physiological function of airway epithelium.MethodsBALB/c mouse tracheal-bronchial epithelial cells were obtained by digestion with 1 mg/mL protease in cold temperature overnight, and the digestion time was optimized to ensure the quantity and viability of the obtained cells. After removing fibroblasts by differential velocity adhesion method, the cells were cultured into collagen coated Transwell inserts. Proliferating phase and air-liquid interface culture were promoted with different culture media.ResultsCell numbers obtained by cold protease overnight digestion for 12 h, 14 h and 16 h were (1.78±0.33)×105, (1.93±0.26)×105 and (2.01±0.28)×105, respectively. Cell viability by trypan blue staining were (96.86±0.25)%, (94.73±1.63)% and (86.87±5.95)%, respectively. Cells were 100% confluent in Transwell chamber after 1-week proliferation, and the ciliary beat frequency was observed under microscope after 2 - 3 weeks of air-liquid interface culture. The cilia structure was confirmed by hematoxylin-eosin staining, electron microscopy and immunofluorescence. Ciliary beat frequency of the cells obtained by this method was consistent with that of mouse trachea in vivo, which further demonstrated its capacity in simulating the physiological function of airway epithelium. ConclusionsThe separation and air-liquid interface culture system as well as the ciliary beat frequency measurement method established in this experiment is simple, stable, efficient and reliable, which establishes a substantial foundation for exploring the pathogenesis and treatment mechanism of airway diseases. It can also provide reference for the culture of epithelium in the airway of other species and/or other organs.