Objective To construct small interfering RNA（siRNA） eukaryotic expression vector specific for human hnRNP K gene，and to observe its silencing effects on hnRNP K gene in A549 cells.Methods The expression vectors of pSUPER/hnRNP K siRNAa，pSUPER/hnRNP K siRNAc and pSUPER/siRNAn were constructed by gene recombination and then transfected into the A549 lung carcinoma cell line by using Lipofectamine2000（a and c respectively represented A and C fragments in hnRNP K coding sequence contained 19 nts，n represented nonsense fragment as control）.The mRNA and protein were harvested after 24 h and analyzed for the expression of hnRNP K by RT-PCR and Western blotting respectively.Results The siRNA vector targeted to hnRNP K successfully decreased hnRNP K mRNA and protein levels 24 h after transfection in A549 cells.Relative expressed doses of hnRNP K mRNA in lung cancer cells transfected by hnRNP K siRNAa and hnRNP K siRNAc respectively were 0.24±0.53 and 0.28±0.57 after 24 h，which were significantly lower than that in the control group（both Plt;0.01）.The gray scale values of hnRNP K protein were 0.23±0.11 and 0.28±0.09 respectively，which were also significantly lower than those in the control group（both Plt;0.05）.And pSUPER/hnRNP K siRNAa was the most effective one.Conclusion Eukaryotic expression vector of siRNA specific for hnRNP K is successfully constructed，which lays the basis for the function study of hnRNP K gene and its application in the treatment of lung carcinoma.
ObjectiveTo understand the obstacles in the practice of pulmonary rehabilitation between doctors and patients.MethodsMedical staff and patients with chronic obstructive pulmonary disease (COPD) in public hospitals in this region were randomly sampled, and a questionnaire survey was conducted on possible obstacles to the practice of pulmonary rehabilitation.ResultsTotal of 265 medical staff and 120 COPD patients were recruited in this survey. The obstacles of pulmonary rehabilitation practice of medical staff in clinical work are poor cooperation of patients and their families (84.2%), medical staff’s insufficient awareness of pulmonary rehabilitation (82.3%), and lack of practice and guidelines (78.9%), lack of objective conditions such as site, equipment and equipment (75.1%), lack of multidisciplinary teams (74.3%), pulmonary rehabilitation has fallen by the wayside (73.6%) etc. The main obstacles for COPD patients in pulmonary rehabilitation are lack of access to relevant knowledge (52.4%), insufficient knowledge (36.5%), inconvenient transportation, economic problems and other objective conditions (33.3%). After multiple correspondence analysis, there are differences in the degree of correlation between obstacle factors and groups with different characteristics.ConclusionsThere are many factors hindering the development of pulmonary rehabilitation and there are certain differences among different populations, but the lack of understanding of pulmonary rehabilitation between doctors and patients is the primary problem. It is necessary to improve the cognition of both doctors and patients on pulmonary rehabilitation, and then to solve the obstacles in the implementation of pulmonary rehabilitation.