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.
Objective To investigate the effect of myeloid derived suppressor cells ( MDSCs) on airway inflammation of asthmatic mice. Methods Five male BALB/ c mice aged 6 weeks were used for preparing 4T1 tumor bearing mice. Thirty female BALB/ c mice aged six weeks were randomly divided into a normal control group, an athmatic model group, and a cell transplantation group. The MDSCs were separated frommyeloid tissue of tumor-bearing mice using amagnetic cell sorting systemand cultured in RPMI medium 1640 containing GM-CSF. The morphologic characteristics of these cells were observed under lightmicroscope and the phenotypic figures were analyzed with flow cytometry. The mice in the model group and the cell transplantation group were sensitized by ovalbumin and then stimulated with nebulized ovalbumin. The mice in the cell transplantation group were intravenously administered MDSCs which purified by magnetic cell sorting system at 10 days after sensitization. The airway inflammation was evaluated by HE staining. The total and differential cell counts in bronchoalveolar lavage fluid ( BALF) were measured.Results The neutrophil and eosinophil infiltration in pulmonary tissue was dramatically increased in the model group, but not observed in the normal control group and was much milder in the cell transplantation group. The total cell count, the eosinophil and lymphocyte counts in BALF of the model group and the cell transplantation group were significantly higher than those of the normal control group( P lt; 0. 05) , and the number of eosinophils in BALF of the cell transplantation group was decreased when compared with that of the model group( P lt;0. 05) . Conclusion MDSCs via intravenous infusion can effectively suppress airway inflammation in a mouse asthma model.
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.
At present, tamponade agent which being used in retinal surgery is mainly sterile air, gas and silicone oil. Sterile air is mostly used in the treatment of simple retinal detachment. Gas or silicone oil as tamponade is greatly applied for complicated retinal detachment. In recent years, with the application of micro-invasive vitrectomy under a wide-angle viewing system and perioperative anti-vascular endothelial growth factor drugs, application of intraocular filling materials also has changed. The application of silicone oil is significantly reduced. Percentage rate of gas as tamponade for retinal detachment is reduced. The application of sterile air as tamponade is rising. With selecting indication carefully and picking up the suitable air or gas, doctor will reduce the workload. It will also reduce the social burden and benefit patients.
ObjectiveTo observe the clinical effect of vitrectomy with internal limiting membrane peeling and air tamponade in the treatment of idiopathic macular hole (IMH).MethodsA prospective, non-randomized controlled case cohort study. Twenty eyes of 20 patients with IMH in Ophthamology Department of The Second Hospital of Jilin University from September 2016 to January 2017 were included in this study. There were 4 males (4 eyes) and 16 females (16 eyes), with the mean age of 60.60±9.50 years and the mean disease course of 9.55±15.5 months. The stage of macular hole ranged from Ⅱ to Ⅳ. All patients underwent BCVA, intraocular pressure, spectral-domain OCT (SD-OCT) and multifocal ERG examinations. The BCVA examination was performed using the international standard visual acuity chart, which was converted into logMAR visual acuity. The minimum diameter (MIN), bottom diameter (BD), height (H), the defective diameter of ellipsoid zone, the defective diameter of external limiting membrane (ELM) of the macular hole were measured by SD-OCT (German Heidelberg). Then the macular hole index (MHI=H/BD), hole traction index (THI=MD/BD) and hole diameter index (DHI=H/MD) were calculated. The response density in P1 wave was detected by mf-ERG. The logMAR BCVA was 1.18±0.54. There were 5 eyes with less than 400 μm MIN, 13 eyes with 400-700 μm MIN and 2 eyes with over than 700 μm MIN. All the patients were underwent vitrectomy with internal limiting membrane peeling and air tamponade. According to the SD-OCT image features after surgery, IMH closure was divided into type Ⅰ closed, type Ⅱ closed and unclosed. The follow-up was equal or greater than 3 months. The closure rate of IMH, BCVA and the changes of microstructure parameters of macular area were observed. Spearman correlation analysis was used to analyze the correlation between BCVA and the wave response density in P1 before surgery and the efficacy after IMH surgery.ResultsClosure rate of IMH: 18 eyes (90.0%) were completely closed (all type Ⅰ closed). The closure rate of macular hole with the MIN less than 400 μm was 100.0% (5/5), 400-700 μm was 92.3% (12/13), and over than 700 μm was 50% (1/2). BCVA: the mean logMAR BCVA at 1 week, 1 month, 3 months after surgery were 0.83±0.54, 0.65±0.41, 0.48±0.34, and there was statistically significant difference between preoperative and postoperative BCVA (t=3.382, 4.459, 5.250; P=0.003, 0.007, 0.004). The outcomes of SD-OCT at 3 months after surgery: 18 eyes (90.0%) with intact ELM. The defective diameter of ellipsoid zone was 260.34±272.08 μm, there was statistically significant difference between before and after surgery (t=13.545, P=0.002). The outcomes of mf-ERG: the P1 wave response density of the ring 1 and ring 2 after surgery were increased compared with before surgery (t=-16.748, -6.151; P＜0.01,＜0.01). The results of Spearman correlation analysis: there was a positive correlation between postoperative BCVA and preoperative MIN, the defective diameter of ellipsoid zone and ELM, and postoperative the defective diameter of ellipsoid zone (r=0.56, 0.59, 0.68, 0.52; P=0.010, 0.006, 0.001, 0.019). The postoperative BCVA was negatively correlated with the P1 wave response density of ring 1 and ring 2 of mf-ERG (r=－0.34, －0.16; P=0.006, 0.020). The IMH closure had a significant negative correlation with MIN and the defective diameter of ELM (r=－0.449, －0.449; P=0.047, 0.047). MHI and THI were positively correlated with the closure of the hole (r=0.474, 0.546; P=0.035, 0.013). Intraocular pressure increased in 2 eyes during the follow-up period, and returned to normal within 1 week after the administration of antihypertensive drugs. There were no complications during or after the operation.ConclusionsVitrectomy with internal limiting membrane peeling and air tamponade in the treatment of IMH is safe and effective. The MIN, MHI THI are significantly correlated with the curative effect after IMH surgery, which could be used as the index to predict and evaluate the curative effect. The response density of ring 1 and ring 2 in P1 wave of mf-ERG is signifi-cantly improved compared with before surgery, and which is negatively correlated with BCVA.
ObjectiveTo evaluate the efficacy and impacts of air tamponadein treating rhegmatogenous retinal detachments (RRD) including inferior breaks after 25G pars plana vitrectomy (PPV).MethodsA prospective non-randomized study. From November 2016 to December 2018, 110 RRD patients receiving PPV in the Ophthalmology Department of Affiliated Hospital of Nantong University were included in the study. All the patients choose the operation mode voluntarily after fully understand the condition and operation mode. There were 90 eyes in the air filling group and 20 eyes in the silicone oil filling group. There was no difference of age (t=－0.082), sex ratio (χ2=1.538), left or right eye (χ2=0.839), average course of disease (Z=－0.276), intraocular pressure (t=3.669), axial length (t=0.765), grade of proliferative vitreous retinopathy (Z=－2.101), high myopia (χ2=0.013), lens state (χ2=1.275), logMAR BCVA (t=－0.681), the scope of retinal detachment (t=0.970), the site (χ2=0.108) and numbers (t=0.158) of the retinal holes, whether involving macula (χ2=1.791) between the two groups (P＞0.05). At 1, 3, and 6 months after surgery, the first retinal reduction rate, BCVA increase rate, visual recovery rate, intraocular pressure, fovea retinal thickness (CFT), ellipsoid and cross sectional area, and the occurrence of postoperative complications in the two groups were observed and compared. Independent sample t test and Chi-squared test, Mann-Whitney U test, and ANOVA were used for data comparision.ResultsAfter the first operation, retinal reattachmnents were achieved in 87 eyes (96.6%) in the air group and 19 eyes (95.0%) in the silicone oil group (χ2=0.130, P＞0.05). In the air group and the silicone oil group, there were 7 and 2 eyes with subretinal holes, whose retinal reattachmnents were achieved. At 6 months after surgery, logMAR BCVA of the eyes in the air group and the silicone oil group were 0.23±0.23 and 0.37±0.23, respectively. All of them were higher than that before operation (t=－2.410, P＜0.001). BCVA in the air group was superior to that in the silicone oil group (P＜0.05). The increase rate of BCVA of the eyes in the air group and the silicone oil group were 0.69±0.28 and 0.48±0.30 logMAR units, respectively (t=－3.225, P＜0.05). The CFT of the eyes in the air group and the silicone oil group were 226.87±42.30 μm and 234.83±36.10 μm, respectively (t=－0.448, P=0.657). In the air group and silicone oil group, posterior cataract was observed in 15 and 4 eyes, subretinal residual fluid in 1 and 2 eyes, and visual object deformation in 3 eyes respectively. In the air group and silicone oil group, posterior cataract was observed in 15 and 4 eyes, subretinal residual fluid in 1 and 2 eyes, and visual object deformation in 3 eyes respectively.ConclusionsAir filling combined with PPV can effectively treat RRD, which is suitable for both superior and inferior holes. The visual acuity recovered well and the complication is less. The time of retinal detachment (especially the time of macular detachment) is an important factor affecting the prognosis.
Objective To investigate the effects of TiotropiumBromide on airway inflammation in a rat model of chronic obstructive pulmonary disease( COPD) . Methods Thirty Wistar rats were randomly divided into three groups. Group A received normal breeding as normal control. Group B and group C received LPS( 200 μg, intratracheally injected at the 1st and the 14th day) and tobacco exposure( from the 2nd day to the 30th day except the 14th day) to establish COPD model. And group C received a nebulized dose of Tiotropium Bromide( 0. 12 mmol / L, 10 minutes) 30 minutes before the tobacco exposure each time. Airway resistance and compliance were measured before sacrificed. Histological examination was performed with Hematoxylin-Eosin staining. The concentrations of IL-8 and LTB4 , total and differential cells counts in bronchoalveolar lavage fluid( BALF) were examined, and the concentrations of IL-8 and LTB4 in blood serum were also examined by ELISA. Results Severe lung inflammation and decreased lung function were demonstrated in the rats in the group B compared with those in the group A. The inflammatory cell counts in BALF, and the levels of IL-8 and LTB4 in BALF and serum were significantly increased in the group B compared with those in the group A. Tiotropium Bromide administration improved the parameters above. Conclusions The results suggest that Tiotropium Bromide can alleviate the lung inflammation and improve the lung function in a rat COPD model. These effects may be exerted through reducing the mediators of inflammation.
Objective To explore the activity of Ca2 + -activated K+ ( KCa) inairwaysmoothmuscle cells( ASMCs) in a rat model of chronic obstructive pulmonary disease( COPD) , and to observe the effect of 11, 12-Epoxyeicosatrienoic acid( 11, 12-EETs) on the KCa channel of ASMCs. Methods Forty male Sprague-Dawley rats were randomly assigned to a COPD group and a normal control group. The rats in the COPD group were exposed to cigarette smoking in a relatively closed chamber to induce COPD. The ASMCs were isolated from small bronchi using an acute enzymatic digestion method. In the symmetrical high K+ solution,the KCa currents were separated with inside-out configuration using the patch clamp technique. The activity of KCa currents in ASMCs between the COPD group and the normal group were compared and the effect of 11, 12-EETs on KCa channel was recorded. The opening probability( Po) , opening time( To) and closing time ( Tc) of the KCa were measured. Results Compared with the normal group, Po of KCa in the COPD rats was much shorter ( 0. 084 ±0. 028 vs 0. 198 ±0. 029, P lt; 0. 01) , To was shorter [ ( 0. 732 ±0. 058) ms vs ( 1. 648 ±0. 152) ms, P lt; 0. 01] and Tc was longer[ ( 12. 259 ±2. 612) ms vs ( 6. 753 ±1. 237) ms, P lt;0. 01] . 11, 12-EETs can evoke the activity of KCa currents of ASMCs in the COPD rats while Po was increased( 0. 227 ±0. 059 vs 0. 084 ±0. 028, P lt; 0. 01) , To was much longer[ ( 2. 068 ±0. 064) ms vs ( 0. 732 ±0. 058) ms, P lt; 0. 01] , and Tc was shorter [ ( 4. 273 ±0. 978) ms vs ( 12. 259 ±2. 612) ms, P lt;0. 01] .Conclusions The results suggest that the decreasing of KCa activity plays an important role in the development of COPD. 11,12-EETs can directly evoke the activity of KCa channel in COPD rats, thus relax the airway smooth muscles.
