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.
Pneumothorax and pulmonary air leakage are common emergency and complication in department of thoracic surgery. The treatment measures of air leakage include conservative observation, needle aspiration, chest tube drainage, surgical operation, and pleurodesis. In selection of treatment methods, there is no unified and feasible standard to choose a reasonable, effective and economic therapeutic regimen. Inapposite treatment will prolong hospitalization days, increase cost, even may lead to death. Therefore, sorting out and summarizing the therapies is vitally important for clinical conduction. We retrieved about 80 reports in PubMed, Sciencedirect, and CNKI while we searched "pneumothorax" or "lung air leakage" and "treatment" as key words or title in recent ten years. We review and summarize the selection and development of them.
Objective To investigate the application of air leak test combined with methylene blue solution leak test in the detection of anastomotic leakage after total mesorectal excision （TME） in rectal cancer. Methods In total of132 patients with rectal cancer underwent Dixon according to TME in our hospital from Mar. 2010 to Mar. 2013 were enrolled. All patients were randomly divided into air leak test group （n=65） and air leak＋methylene blue solution leak test group （n=67）. The intestinal anastomosis of patients in air leak test group were clamped at 2 cm from the upper endof bowel， then injecting 500 mL distilled water to pelvic， and placing 24# Foley catheter through the anus. The catheter balloon was injected with water to close anus， and then injected with 50 mL gas to find the anastomotic leakage where bubbles happened， and then repaired it. Patients of air leak＋methylene blue solution leak test group were treated with methylene blue solution test in addition. After sucking out of the distilled water in pelvic and gas in the rectum， 1 bottle of methylene blue solution （20 mg） and 50 mL saline were injected， observing the location where the methylene blue solutionleaking out and repaired it. Results Three cases （4.62%） of anastomotic leakage were found during operation in air leak test group， and 9 cases （13.85%） were found after operation. Of the 9 cases， 5 cases were cured with placement of adeq-uate drainage and symptomatic treatment， 3 cases were cured with anal patch， and 1 case was cured with transverse colon fistula and drainage. In total of 15 cases （22.39%） were found anastomotic leakage， 2 cases of them were found by air leak test and another 13 cases were found by methylene blue solution leak test during operation in air leak＋methylene bluesolution leak test group， but no one suffered anastomotic leakage after operation. Compared with air test group， detectionrate of anastomotic leakage during operation was higher （P＜0.05）， and incidence rate of anastomotic leakage after opera-tion was lower in air leak＋methylene blue solution leak test group （P＜0.05）. Conclusions Large anastomotic leakage can be found by using air leak test， and small and hidden leakage can be found by using methylene blue solution leak test， combination method of the two experiments is better. Repair can be performed effectively under direct vision.
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 observe the effects of astaxanthin (AST) on the airway inflammation and remodeling in the asthmatic rats. Methods Fifty male Wistar rats were randomly divided into five groups (n=10 for each group): saline-sensitized and-saline-challenged group (the control group), bronchial asthma group (the asthma group), bronchial asthma+astaxanthin 5 mg/kg gavage treatment group (the AST 5 mg/kg group), bronchial asthma+10 mg/kg gavage treatment group (the AST 10 mg/kg group), and bronchial asthma+50 mg/kg gavage treatment group (the AST 50 mg/kg group). The level of interleukin-5(IL-5), interleukin-13(IL-13), interferon-γ(IFN-γ), tansforming growth factor-β (TGF-β), malondialdehyde (MDA) and superoxide dismutase (SOD) in the bronchoalveolar lavage fluid (BALF) and the total IgE level in the serum were measured using enzyme linked immunosorbent assay (ELISA).The infiltration of airway inflammatory cells and the degree of airway epithelial cells detachment, the extent of goblet cell hyperplasia and the severity of subepithelial collagen deposition were evaluated on the hematoxylin eosin (HE), periodic acid Schiff (PAS) and Masson trichrome stained lung sections. reverse transcription-polymerase chain reaction (RT-PCR) was used to measure the expression of mucin 5A and C (MUC5AC) messenger ribonucleic acid(mRNA) in lung tissue; Immunohistochemical staining was used to determine the expression of MUC5AC protein in the rat airway epithelium. Results The level of IL-5, IL-13, TGF-β, MDA and the total IgE in the serum respectively [(36.73±2.29), (53.99±2.70), (60.89±2.54)ng/mL,(18.65±0.76)umol/L, (54.50±2.91)ng/mL], the extent of inflammatory cells infiltration (46.24 ± 4.26), the extent of eosinophils infiltration (2.09± 0.13), the extent of epithelial cells detachment [(6.09±0.45)%], the extent of goblet cell hyperplasia [(13.65±1.90)%], the extent of subepithelial collagen deposition [(17.58±2.14)%], the MUC5AC mRNA expression level, and the lung tissue MUC5AC protein expression IOD value (187±12) in the asthma group were all higher than those in the control group (P<0.01 or P<0.001), the level of IFN-γ and SOD in the BALF[(26.38±1.70) ng/mL], [(16.37±1.22) U/L], was lower than that in the control group (P<0.001); The level of IL-5, IL-13, total IgE, TGF-β, MDA, the inflammatory cells infiltration in the airway epithelial, the degree of epithelial cell damage and detachment, the degree of goblet cell hyperplasia, the degree of subepithelial collagen deposition, the MUC5AC mRNA expression in lung tissue,and the MUC5AC protein expression in airway epithelial cells in the AST treated groups were all lower than those in the asthma group (P<0.05 or P<0.01 or P<0.001),the level of IFN-γ, SOD in the BALF was higher than that in the asthma group (P<0.05 or P<0.01). Conclusion Astaxanthin can inhibit airway inflammation, downregulate airway MUC5AC expression, inhibit goblet cell proliferation, and alleviate airway remodeling in rats with bronchial asthma.
Lung transplantation has been the only valid method in treating end-stage lung diseases, airway complications are the main cause to the failure of surgery and common postoperative complications. With the development on patient selection, organ preservation, surgical technique, immunosuppressive therapy and postoperative surveillance, the successful ratio of surgery has become most satisfactory. However, airway complications are still common after lung transplantation. Among these, the airway anastomosis stenosis is more predominant than others. The living quality and long-dated survival rate are highly improved by paying enough attention to the formation,corresponding management for tracheal stenosis. The progress of the cause, prevention and treatment of airway anastomosis stenosis after lung transplantation is reviewed in this article.
【Abstract】 Objective To study the role of house dust mite ( HDM) induced airway epithelium TLR4 expression and T lymphocyte activation in asthmatic inflammation. Methods Thirty BALB/ c mice were randomly divided into an ovalbumin ( OVA) group, a HDMgroup, and a control group. The mice in the OVA group were sensitized with OVA and Al( OH) 3 , and repeatedly exposed to aerosolized OVA. The mice in the HDMgroup and the control group were sensitized and challenged with HDMand saline, respectively.Histopathology changes of pulmonary tissue and airway were observed under light microscope. Levels of IL-4, IL-5, IL-13, IL-17, and IFN-γin BALF were measured by ELISA. Total and differential cell counts in bronchoalveolar lavage fluid ( BALF) were also measured. The mRNA and protein expressions of TLR4 weredetected by quantitative real-time PCR and Western blot, respectively. Th1, Th2, and cells in the peripheral blood were detected by flow cytometry. Results Light microscope revealed eosinophil specific inflammatory cells infiltration around the peribronchovascular region,mucus gland hyperplasia, and airway mucous plug inthe OVA group. The HDM group showed more severe alveolar and intersititial congestion and neutrophils infiltration. The control group showed intact alveolus with few mucous plug and inflammatory cells.Compared with the OVA group, significant increases in the number of total cells and neutrophils, as well as significantly higher expression of IL-4, IL-5, IL-13, and IL-17 were detected in the HDMgroup ( P lt;0. 05) ,while IFN-γexpression had no significant change ( P gt;0. 05) . The expression of TLR4 mRNA and protein significantly increased in the HDMgroup( P lt; 0. 05) , and did not change significantly in the other two groups ( P gt;0. 05) . The percentages of Th2 and Th17 cells in peripheral blood in the HDMgroup were significantly higher than the OVA group ( P lt;0. 05) . Conclusion HDM may induce inflammatory cells infiltration andactivation of Th2 and Th17 lymphocyte cells via up-regulation of TLR4 expression in airway epithelium,which might play an important role in asthmatic inflammation.
ObjectiveTo monitor the airway inflammatory factors in exhaled breath condensate(EBC) of severe stable COPD patients during salmeterol/fluticasone (50/500μg, bid) treatment, and explore their clinical significance. MethodsTwenty-four sever stable COPD patients and 18 healthy controls were included in the study. EBC was collected from COPD patients before treatment (day 0) and 14 days, 28 days, 90 days after treatment. Meanwhile lung function test and SGRQ score were measured.Concentrations of IL-6 and IL-10 were measured by liquid chip and 8-isoprostane by enzyme-linked immunosorbent assay. ResultsLevels of 8-isoprostane, IL-6 and IL-10 in EBC were significantly higher in the sever stable COPD patients before treatment compared with the healthy controls. 8-isoprostane was decreased significantly at day 14 compared with day 0[(11.59±4.12) pg/mL vs. (14.17±4.66) pg/mL, P < 0.05], and kept in low level till day 90 (P > 0.05). IL-6 was significantly decreased at day 28 compared with day 0[(1.46±0.19) pg/mL vs. (1.59±0.19) pg/mL, P < 0.05], but did not change significantly till day 90. IL-10 was in low level but showed increase at day 90 compared with day 28[(1.72±0.19) pg/mL vs. (1.62±0.12) pg/mL, P < 0.05]. FEV1 and FEV1/FVC were improved and SGRQ score was decreased after 90 days treatment (P < 0.05). FEV1 was not correlated with 8-isoprostane, IL-6 or IL-10 level. ConclusionsDynamic observation of EBC 8-isoprostane level in severe COPD patients can help in evaluating drug efficacy. IL-10 may play a role in airway anti-inflammation.
Objective To investigate the effects of smoking intensity, duration and cessation on mRNA and protein expressions of matrix metalloproteinase-9 ( MMP-9) in tracheal epitheliumof rats, and the relationship between smoking or smoking cessation and airway remodeling in chronic obstructive pulmonary disease ( COPD) . Methods Forty Wistar rats were randomly divided into 5 groups, ie. a normal control group, a long termheavy smoking group, a short termheavy smoking group, a long termlight smoking group,and a smoking cessation group which was exposed to room air for 10 weeks after long term heavy smoking.The expressions of MMP-9 mRNA and protein in tracheal epithelium of rats were detected by in situ hybridization and munohistochemistry respectively. Results ( 1) The pathological changes of emphysema were observed in the lung tissue of every smoking rat, and were most sever in the long term heavy smoking group. ( 2) Compared with the normal control group [ ( 0. 88 ±0. 88) PU, ( 2. 80 ±1. 66) PU] , the expressions of MMP-9 mRNA and proteins in tracheal epithelium were remarkable elevated in the long term heavy smoking group [ ( 22. 01 ±2. 86) PU, ( 20. 81 ±2. 46) PU] , the short term heavy smoking group [ ( 14. 94 ±3. 46) PU, ( 13. 68 ±2. 00) PU] , the long term light smoking group [ ( 6. 92 ±2. 71) PU,( 8. 84 ±1. 80) PU] and the smoking cessation group [ ( 19. 00 ±3. 36) PU, ( 14. 82 ±1. 74) PU] ( P lt;0. 01) . Compared with the long term heavy smoking group, the expressions of MMP-9 in tracheal epithelium were decreased in other three smoking groups ( P lt; 0. 05) . Conclusions Smoking could increase the expression of MMP-9 in tracheal epithelium and cause trachea damage and remodeling with intensity and duration in rats. Smoking cessation could decrease the MMP-9 expression and alleviate trachea remodeling,suggesting its role in the prevention of COPD.
Objective To explore the effects of prolonged inhalation of Aspergillus fumigatus ( Af)spores on epithelial cell injury and expression of epidermal growth factor receptor( EGFR) in airways of asthmatic rats. Methods 64 male Wistar rats were randomly divided into 8 groups, ie. chronic asthma group ( group A) , chronic asthma plus Af spores inhalation for 1 week ( group B) , 3 weeks ( group C) and 5 weeks ( group D) , chronic asthma plus saline inhalation for 5 weeks ( group E) , OVA-sensitized and salinechallenged group ( group F) , and OVA-sensitized and saline-challenged plus Af spores inhalation for 5 weeks ( group G) ( each n =8) . The airway resistance ( Raw) and change of Raw after acetylcholine provocation were detected using a computerized system. The concentrations of epidermal growth factor ( EGF) andtransforming growth factor alpha( TGF-α) in BALF were measured by ELISA. The extents of epithelial cell injury and goblet cell hyperplasia were evaluated on hematoxylin and eosin-stained( HE) and periodic acidschiff ( PAS) stained lung sections. The expression of EGFR in airway epithelia was demonstrated byimmunohistochemistry, and the level of EGFR protein in the rat lung tissues was measured by western blot.Results The concentration of EGF( pg/mL) ( 51. 72 ±8. 54, 68. 12 ±7. 85, 86. 24 ±9. 12, respectively)and TGF-α( pg/mL) ( 55. 26 ±9. 30, 75. 58 ±11. 56, 96. 75 ±14. 66, respectively) , detached/ inner perimeter of epithelium( % ) ( 11. 25 ±3. 12, 26. 45 ±5. 56, 28. 50 ±7. 50, respectively) , the ratio of goblet cell area to epithelial cell area ( % ) ( 16. 42 ±5. 24, 22. 64 ±6. 82, 36. 38 ±9. 21, respectively) , the integrated optical density ( IOD) of EGFR positive stain in airway epithelial cells ( 82 ±15,120 ±19, 165 ±21, respectively) , and the EGFR protein levels in lung tissues ( 0. 91 ±0. 26, 1. 61 ±0. 52, 2. 52 ±0. 78,respectively) in group B, C, and D were higher than those in group A, E, F and G( P lt; 0. 05 or P lt;0. 01) .The change rates of Raw( % ) ( 61. 91 ±5. 26, 84. 69 ±6. 38) in group C and D were higher than those in group A, E, F and G ( P lt; 0. 05 or P lt;0. 01) . The IOD of EGFR was positively correlated with detached/inner perimeter of epithelium( % ) and the ratio of goblet cell area to epithelial cell area( % ) ( r = 0. 692,P lt;0. 01; r = 0. 657, P lt; 0. 01, respectively) . Conclusion Prolonged inhalation of Aspergillus fumigatus spores can aggravate airway epithelial cell injury, up-regulate the expression of EGFR in airway epithelial cell and induce goblet cell hyperplasia, thus increase the airway responsiveness in rats with chronic asthma.