Objective To explore the clinical role of transbranchial needle aspiration(TBNA) in the lymph node staging of lung cancer.Methods To forty-six cases of lung cancer with suspected lymph node metastasis by chest CT scan,the TBNA was performed before operation.The cytological results were compared with post-operative pathology.Results In total 94 groups mediastinal nodes,fifty-nine groups were positive by TBNA,sixty-seven groups were diagnosed lymph node metastasis by pathology,the positive accuracy was 88% with the total coincidence rate of 82.6% by cytolagical typing.The overall accurate rate of c-N by TBNA was 86.9%(40/46) compared with p-N.Conclusion TBNA for lymph node staging and pathologic typing of lung cancer is a safe,simple and economic method with high accuracy and high clinical application value.

Objective To establish a real‐time fluorescent polymerase chain reaction and detect 16S rRNA gene of Legionella strains isolated from sputum specimens of patients with pulmonary infection by using this method .Methods 16s rRNA gene of Le‐gionella was used to design primers and probes .The reaction system and reaction conditions were optimized and the specificity ,sen‐sitivity and repeatability of this method were verified by detecting Legionella pneumophila ,non‐Legionella pneumophila and other bacteria .A total of 557 sputum specimens of patients with pulmonary infection were detected ,and PCR‐digestion identification method was carried out as control .Otherwise ,sequences of 16S rRNA were verified in patients with positive detection results .Re‐sults The results showed that all reference strains of Legionella were positive ,while all of other bacteria were negative ,and the sensitivity was 102 CFU/mL .Among sputum specimens collected from 577 cases of patients with pulmonary infection ,the positive rate of Legionella detected by using real‐time fluorescent PCR and PCR‐digestion identification method was 23 .1% and 19 .9% re‐spectively ,while the positive rate was 17 .2% by verifying the sequences of 16s rRNA .There were no statistically significant differ‐ences of positive rate among the three methods(P>0 .05) .Conclusion The real‐time fluorescent PCR is fast and convenient in de‐tection of Legionella strains isolated from sputum specimens of patients ,which could be an assisted method for clinically diagnosing Legionella infection .

Objective : The CRISPR / Cas9 technology was applied to construct PDE4D homozygous knockout mice to provide a basis for in-depth investigation of PDE4D gene function and mechanism of action． Methods: A vector was constructed for PDE4D gene exon 4，5 microinjected into fertilized eggs of C57BL /6J mice，and PDE4D －/ - mice were obtained after maternal breeding and offspring mating，and the mice genotypes were determined by PCR product sequencing and genotype identification techniques．Changes in morphology and function of the major organs of the mice were detected using an ultrasound imaging system and H＆E staining，and the expression of PDE4D protein in the mice was verified by Western blot assay. Results : The PDE4D －/ － mouse genotype was stably inherited， the mice were small，and there were no obvious morphological and histological changes in the major organs in vivo. The PDE4D expression was reduced or largely absent in the major tissues of PDE4D heterozygous or pure knockout mice，and the knockout effect was better． Conclusion PDE4D －/ － mice were successfully established using CRISPR / Cas9 technology，and no significant physiological abnormalities were found，which could be used for disease pathogenesis and drug research using PDE4D as the target.