ObjectiveTo explore and analyze the effect of simulation-based learning combined with debriefing in neonatal resuscitation training.MethodsA total of 114 clinical medical staffs attended the neonatal resuscitation training course hold by Department of Neonatology, Quzhou Maternal and Child Health Hospital from November 2014 to May 2015, and were randomly assigned to observation (n=60) and control group (n=84) by coin tossing. Staffs in the observation group adopted to training skills with simulation-based learning combined with debriefing,while those in the control group were educated with traditional method. The examinations on theoretical knowledge were taken before and after the training. Operational exam and self-confident questionnaire for all staffs on each procedure taught in the course were taken at last. Scores of the exams and self-confident questionnaire were compared between the two groups witht-test and Mann-WhitneyU test.ResultsThe mean score of theoretical test rose up significantly after the training in both observation and control group (25.19±2.62 vs 20.17±3.71,t=7.725,P<0.01; 25.44±2.64 vs 18.90±4.27,t=11.170,P<0.01), but no difference was found in this score after the training between the two groups (t=0.492,P=0.624). The practical operation examination score in the observation group was higher than that in the control (34.05±1.34 vs 31.32±4.10,t=4.183,P<0.01). All questionnaires sent to the staffs were retrieved (100%), and the total values after the training in the observation group were higher than in the control (mean rank: 92.81 vs 57.99; rank sum:5 569 vs 4 872,Z=-4.96,P<0.01).ConclusionsSimulation-based learning combined with debriefing is a much more effective teaching methods for neonatal resuscitation training, which might quickly improve the resuscitation skills of clinical staffs.

PURPOSE: This study aimed to identify potential epidermal growth factor receptor (EGFR) gene mutations in non-small cell lung cancer that went undetected by amplification refractory mutation system-Scorpion real-time PCR (ARMS-PCR). MATERIALS AND METHODS: A total of 200 specimens were obtained from the First Affiliated Hospital of Guangzhou Medical University from August 2014 to August 2015. In total, 100 ARMS-negative and 100 ARMS-positive specimens were evaluated for EGFR gene mutations by Sanger sequencing. The methodology and sensitivity of each method and the outcomes of EGFR-tyrosine kinase inhibitor (TKI) therapy were analyzed. RESULTS: Among the 100 ARMS-PCR-positive samples, 90 were positive by Sanger sequencing, while 10 cases were considered negative, because the mutation abundance was less than 10%. Among the 100 negative cases, three were positive for a rare EGFR mutation by Sanger sequencing. In the curative effect analysis of EGFR-TKIs, the progression-free survival (PFS) analysis based on ARMS and Sanger sequencing results showed no difference. However, the PFS of patients with a high abundance of EGFR mutation was 12.4 months [95% confidence interval (CI), 11.6−12.4 months], which was significantly higher than that of patients with a low abundance of mutations detected by Sanger sequencing (95% CI, 10.7−11.3 months) (p < 0.001). CONCLUSION: The ARMS method demonstrated higher sensitivity than Sanger sequencing, but was prone to missing mutations due to primer design. Sanger sequencing was able to detect rare EGFR mutations and deemed applicable for confirming EGFR status. A clinical trial evaluating the efficacy of EGFR-TKIs in patients with rare EGFR mutations is needed.
Adenocarcinoma/genetics ; Adenocarcinoma/pathology ; Aged ; Aged, 80 and over ; Animals ; Base Sequence ; Disease-Free Survival ; Female ; Humans ; Lung Neoplasms/genetics ; Lung Neoplasms/pathology ; Male ; Middle Aged ; Mutation/genetics ; Mutation Rate ; Real-Time Polymerase Chain Reaction/methods ; Receptor, Epidermal Growth Factor/genetics ; Sequence Analysis, DNA/methods ; Treatment Outcome

Proximity labeling catalyzed by promiscuous enzymes,such as APEX2,has emerged as a powerful approach to characterize multiprotein complexes and protein-protein interactions.How-ever,current methods depend on the expression of exogenous fusion proteins and cannot be applied to identify proteins surrounding post-translationally modified proteins.To address this limitation,we developed a new method to label proximal proteins of interest by antibody-mediated protein A-ascorbate peroxidase 2(pA-APEX2)labeling(AMAPEX).In this method,a modified protein is bound in situ by a specific antibody,which then tethers a pA-APEX2 fusion protein.Activation of APEX2 labels the nearby proteins with biotin;the biotinylated proteins are then purified using streptavidin beads and identified by mass spectrometry.We demonstrated the utility of this approach by profiling the proximal proteins of histone modifications including H3K27me3,H3K9me3,H3K4me3,H4K5ac,and H4K12ac,as well as verifying the co-localization of these iden-tified proteins with bait proteins by published ChIP-seq analysis and nucleosome immunoprecipi-tation.Overall,AMAPEX is an efficient method to identify proteins that are proximal to modified histones.