Objective To explore the influence of polyamine-cholesterol cationic liposome (PCL)-mediated CpGODN aerosol on eosinophiles in the lung tissue of mouse asthma model. Methods Mouse asthma model was replicated by challenging with 1% ovalbumin aerosol. Mice were categonied into four groups, namely normal control, asthma control, CpGODN/PCL treatment group and CpGODN treatment group (6 each). The left lungs of mice were harvested, serially sectioned, hematoxylin and eosin (HE)-stained, and the infiltration of eosinophiles (EOS) was examined under microscope. Meanwhile, the bronchoalveolar lavage fluid (BALF) was collected for total and eosinophil cells count. Results An ovalbumin challenged mouse asthma model was successfully replicated. Pathological observation of the lung of asthma control showed increase in mucous secretion in alveolar space and peribronchial infiltration of large amount of inflammatory cells, primarily EOS and lymphocytes. The total cell number, EOS number and the ratio in BALF were significantly higher in asthma control group compared with that in both normal control group and CpGODN treatment group (P

Objective To evaluate the significance of fibreoptic bronchoscopy combined with percutaneous pneumocentesis in the diagnosis of peripheral pulmonary lesions.Methods 269 patients with peripheral pulmonary diseases were diagnosed with fibreoptic bronchoscopy biopsy combined with bronchial brush biopsy.In 89 out of the 269 patients no positive findings were obtained,because the lesions were located in the lung hilum,medial sphere or mesosphere,when X-ray guided fibreoptic bronchoscopy biopsy combined with bronchial brush biopsy were employed.In 77 out of the 89 patients positive fingings were still lacking,as their lesions were located in the mesosphere or peripheralzone,when X-ray guided percutaneous pneumocentesis biopsy was used.In 70 out of the 77 patients,positive fingings were lacking,as their lesions were just beneath the chest wall,when B-ultrasonography guided percutaneous automatic elastic biopsy was employed.Results 276 times of fibreoptic bronchoscopy biopsy combined with bronchial brush biopsy were performed in 269 patients,final diagnosis was obtained in 82 of them,with a diagnostic rate of 30.5%;94 times of X-ray guided fibreoptic bronchoscopy biopsy combined with bronchial brush biopsy were performed in 89 patients,and final diagnosis was obtained in 66 patients,with a diagnostic rate of 74.2%;85 times of X-ray guided percutaneous pneumocentesis biopsy were performed in 77 patients,final diagnosis was obtained in 52,with a diagnostic rate of 67.5%;79 times of B-ultrasonography guided percutaneous automatic elastic biopsy were performed in 70 patients,final diagnosis was obtained in 49,with a diagnostic rate of 70.0%.A final diagnosis was made in a total of 249 patients,with a diagnostic rate of 92.6% when 4 methods were used in combination.Conclusion Combined use of fibreoptic bronchoscopy with percutaneous pneumocentesis biopsy may remarkably raise the diagnostic rate in the patients with peripheral pulmonary diseases,and is worth to be recommended for general use in clinical work.

Objective To investigate the efficacy and safety of interventional therapy for central airway stenosing pulmonary disease. Methods In 389 petients diagnosed as central airway stenosis, their primary diseases were handled timely and correctly. On the basis of handling the primary disease, cetral airway stenosis was treated by interventional techniques, including electrosurgical unit (ESU), laser, argon plasma coagulation, freezing, high-pressure balloon expansion, stent implantation, local injection and toilet with lavage. Following the American College of Chest Physicians (ACCP) guidelines, the therapeutic effects were evaluated based on correction of stenosis after interventional treatment and the dyspnea scores. Results Of the 389 patients, 154 were treated with electrosurgical unit under bronchoscopy, 18 with laser, 76 with freezing, 69 with argon plasma coagulation, 35 with high-pressure balloon expansion, 21 with tracheobronchial stent implantation, and 16 with local injection and laving clearance. Evaluation of the therapeutic effects was made one month after the interventional treatment. As a result of the interventional treatment, a complete relieve was observed in 152 cases (39.1%), partial relieve in 138 cases (35.5%), mild response in 67 cases (17.2%), no response in 32 cases (8.2%), and with 17 deaths (4.4%). The total effective rate was 91.8%. The dyspnea scores were 1.8?0.5 before the treatment, and 0.7?0.4 after the treatment, with statistically significant difference (P

Objective To investigate the pathological features of airway remodeling and evaluate interventional effects of nerve growth factor (NGF) on the expression of matrix metalloproteinase-9 (MMP-9), an important mediator of airway remodeling, in guinea pig asthma model. Methods Guinea pigs were randomly divided into the control group, asthma group, and antibody NGF intervention group (each group had 8 guinea pigs). In the asthma group the animals were sensitized by repeated exposure to aerosolized ovalbumin combined with Al(OH)_3. The thickness of the smooth muscle of intrapulmonary bronchi was measured by image analysis system. The expression of MMP-9 in bronchi and lung tissues were assayed with immunohistochemistry combined with the micro-image analysis system. The levels of MMP-9 mRNA in bronchi and lung tissues were determined by RT-PCR. Results After repeated allergen challenge, obvious infiltration of inflammatory cells and proliferation of goblet cells and smooth muscle were demonstrated in guinea pig bronchi. Expression levels of MMP-9 in the epithelial cells of bronchi were significantly higher in asthmatic animals than those of control group animals. Compared with asthmatic group, there was mild inflammation reaction, and decrease in collagen deposition and expression of MMP-9 in antibody NGF group animals, and they were not significantly higher than that in control group animals. Conclusions Repeated exposure of allergen induced airway inflammation and remodeling. MMP-9 plays an important role in airway remodeling. Antibody NGF intervention could inhibit airway remodeling through inhibition of the expression of MMP-9.

Objective To evaluate the transfection efficiency and cytotoxicity of polyamine cationic liposomes (PCL), and to screen for a cationic liposome with high potency and low toxicity. Methods Four kinds of PCLs, namely PCL-A, PCL-B, PCL-C and PCL-D, were prepared with polyamine cationic lipid TC-Chol or DC-Chol and the neutral phospholipid DOPE in different molar ratios (1∶1 or 3∶1). The morphology of the liposome was observed by transmission electron microscope. Plasmid PIRES2-EGFP, which contained an enhanced green fluorescencent-protein (EGFP) reporter gene, was transfected into Hela cells or Hep2 cells with PCL-A, PCL-B, PCL-C and PCL-D, respectively. Expression of EGFP was examined by fluorescence microscopy and the cytotoxicity of the transfected cells was determined with MTT assay. Results The shape of PCL-A, PCL-B, PCL-C and PCL-D was round or oval, most of the PCLs were about 50-200nm in diameter. The transfection efficiency of the four kinds of PCLs was about 19.8%-42.1%. Among them, PCL-C prepared with TC-Chol and DOPE at a molar ratio of 3∶1 showed a higher transfection efficiency (39.5%-42.1%) and lower cytotoxicity (cellular survival rate was 86.57%?3.23%-84.43%?4.18%). Conclusions The polyamine cationic liposome prepared with polyamine cationic lipids TC-Chol or DC-Chol and the neutral phospholipid DOPE at a certain molar ratio mediated the transfection of the plasmid PIRES2-EGFP into mammalian cells. PCL-C exhibited higher transfection efficiency and lower cytotoxicity, and it might be a promising agent for gene transfection and gene therapy.

Objective To explore the diagnostic efficacy of bronchoscopy combined with T-SPOT in tuber-culosis. Methods Totally 248 highly suspected tuberculosis patients were included from February 2015 to April 2016 in our department. According to the different sources of sputum specimens ,they were divided into control group,atomization group and bronchoscope group;Depending on the mode of examination,they were divided into bronchoscopy group,T-SPOT group,bronchoscopy+T-SPOT group.To compare anti-acid smear positive rate,tuber-culosis diagnosis and diagnostic efficacy in each group and safety of bronchoscopy. Results The rate of sputum specimens smear-positive was the highest(37.173%) in bronchoscopy.There were statistically significant of the three groups(P=0.001);The highest diagnostic rate of pulmonary tuberculosis was(92.670%)in Bronchoscopy+T-SPOT group,three groups were statistically significant(P=0.000). The specificity,sensitivity,positive predic-tive value,negative predictive value,positive likelihood ratio and Jorden index were highest in diagnosing tuberculo-sis of bronchoscopy+T-SPOT group ,negative likelihood ratio was lowest. There were Complications of three cases of bleeding,one case of pneumothorax,one case of arrhythmia in bronchoscopy group. Conclusion Bronchoscopy combined with T-SPOT can improve the diagnostic efficacy of tuberculosis ,safe and reliable.

Objective To investigate the influence of S100A6 gene RNA interference on the biological behaviors of A549 lung adenocarcinoma cells.Methods The S100A6 gene RNA interference vector was transfected in A549 lung adenocarcinoma by lentivirus.The experiment was divided into three groups:pLenR-GPH group (the vector without S100A6 RNAi gene was transfected),negative control group (no vectors was transfected),and RNAi group (the vector with S100A6 RNAi gene was transfected).S100A6 mRNA and protein were detected using real-time PCR and Western blotting.The biological behavior including cell proliferation,invasion,cell cycle and cell apoptosis were detected by 3-(4,5-dimethyl-2-thiazoly)-2,5-diphenyl-2H-tetrazolium bromide,transwell,and flow cytometer,respectively.Results The expression of S100A6 mRNA of A549 lung adenocarcinoma cell line in RNAi group (0.009 ± 0.001) was significantly decreased than those in negative control group (0.049 ± 0.005) and pLenR-GPH group (0.030 ± 0.006),with statistically significant differences (t =57.56,P =0.000;t =48.21,P =0.000).The expression of S100A6 protein of A549 lung adenocarcinoma cell line in RNAi group (0.107 ± 0.002) was significantly decreased than those in negative control group (0.341 ± 0.005) and pLenR-GPH group (0.311 ± 0.006),with statistically significant differences (t =37.34,P =0.000;t =27.51,P =0.001).The ability of cell proliferation at 48 hours in RNAi group (0.230 ± 0.008) was significantly declined than those in negative control group (0.292 ± 0.038) and pLenR-GPH group (0.307 ± 0.013),with statistically significant differences (t =25.31,P =0.003;t =29.42,P =0.001).The number of transmembrane cells in RNAi group (11.40 ± 1.36) was significantly declined than those in negative control group (26.80 ± 1.83) and pLenR-GPH group (25.80 ± 1.93),with statistically significant differences (t =29.44,P =0.001;t =23.17,P =0.005).The cell proportion of S phase in RNAi group (28.26％ ± 0.38％) was significantly lower than those in pLenR-GPH group (44.73％±0.66％) and negative control group (45.15％ ± 1.69％),with statistically significant (t =63.69,P=0.000;t =71.55,P =0.000).Cell propotion of G2-M phase in RNAi group (26.99％ ± 0.29％) was signi-ficantly higher than those in negative control group (13.26％ ±0.49％) and pLenR-GPH group (12.41％ ± 0.46％),with statistically significant (t =56.31,P =0.000;t =51.39,P =0.000).The cell apoptosis proportion in RNAi group (8.90％ ±0.48％) was significantly higher than those in negative control group (5.84％ ±0.21％) and pLenR-GPH group (5.99％ ±0.37％),with statistically significant (t=51.34,P =0.000;t =47.27,P =0.000).Conclusion S100A6 gene involves the proliferation,invasion,cell cycle and apoptosis of tumor cells,which has close correlation with occurrence,development and metastasis of lung adenocarcinoma.S100A6 gene is hopeful to become a molecular target for the diagnosis and treatment of lung adenocarcinoma.

Aim To investigate the changes of plasticity and distribution in the lungs of asthmatic guinea pigs.Methods Guinea pigs were divided into asthma group and control group.Immunohistochemistry was used in this study to observe the changes of ERK1/2 and Elk phosphoryiation in the lungs of guinea pigs.Results There were significant changes in distribution of EPK1/2 and Elk in the lungs of asthma group.The positive cells were detected on the walls of respiratory bronchioles and alveolar ducts,especially on the smooth muscle layer and basement membrane.In addition,the numbers of positive cell were clearly increased in asthma group(P< 0.01). Conclusion There is close relationship between the EPK1/2 and Elk phosphoryiation and attack of asthma.It may be a reason for persistence and progress of asthma.

Objective To explore the diagnostic value of virtual bronchoscopic navigation (VBN) for peripheral pulmonary lesions. Methods 200 cases with peripheral pulmonary lesions (0.8 cm < diameter < 4.0 cm) were divided into VBN and control group from June 2014 to June 2015. VBN group: VBN guided ultrafine bronchoscope to the target bronchus, control group: ultrafine bronchoscopy with chest CT as a reference guided to the target bronchus. Results There were no significant differences in the diagnosis rates between VBN group and control group in 200 patients (χ2 = 3.31, P = 0.069); But, the diagnosis rates with diameter more than 2.0 cm and less than or equal to 2.0 cm had statistically significant differences of VBN group and control group (χ2 = 13.45, 5.31, P = 0.000, 0.021, respectively); We also found that the mean time of biopsy tool reach to the lesions had significant differences between the two groups (P = 0.047). There were no significant differences in total checking time and complications (P = 0.230, 0.960, respectively). Conclusions Virtual bronchoscopic navigation did not improve the diagnosis rate of pulmonary peripheral lesions, but shortened the time to locate the lesions.

Objective To explore the security and its influencing factors on benign airway stenosis treated with interventional of high pressure balloon expansion catheter.Methods Clinial data of 39 cases of inpatients suffered from benign airway stenosis were chosen.17 cases were male,and 22 cases were female.The ages of them ranged from 15 to 83 years old.According to the clinical symptoms,HRCT 3D reconstruction,and the results of bron-choscope,all patients were treated with balloon expansion catheter at different criterions.The balloon catheter with size that slightly smaller than the targeted normal bronchial tube was chosen,expansion for average 1 -4 times,single balloon expansion time ranged from 0.5 to 4 min,the pressures were kept at 3 -6 atmosphere,and the highest pres-sure did not exceed 8 atmospheric pressure.The efficacy and complications were retrospectively analyzed.Results 19 cases were completely effective,14 cases were basically effective,6 cases were completely ineffective,and the total effective rate was 84.6% (33 /39 ),the incidence of complications was 35.8% (14 /39 ),moreover,no deaths occurred.Conclusion High pressure balloon catheter expansion is one of commonly used technology in breathing interventional treatment;it has the characteristics of easy operation,and immediate curative effect,and so on.But if the improper operation,incorrect selection of the case,or inaccurate evaluation of the stenosis during operation,serious complications and unnecessary iatrogenic injury can be occurred.Therefore,it is worthy of attention and further summarizing by breathing interventional physicians.