Objective To construct the eukaryotic expression plasmid containing human epidermal growth factor(hEGF) gene with signal peptide(SP).Methods After two pairs of primers were designed and synthesized,the cDNA fragment of hEGF and SP genes were amplified from total RNAs. The amplified cDNA fragments were cloned into pGEM-T vector.The expression plasmids were verified by double endonuclease digestion and DNA sequence analysis. Results With RT-PCR using two pairs of primers,two bands(about 90bp and 180bp) were obtained and confirmed as signal peptide and EGF cDNA fragment with electrophoresis analysis and DNA sequencing after cloned into pGEM-T vector.The SP and EGF cDNA fragments were inserted into plasmid pcDNA3.1(+).The bands of 240bp and 5.4kb were obtained and identified as the full length of SP-EGF cDNA fragment by DNA sequence analysis.Conclusion The eukaryotic expression plasmids containing hEGF gene is successfully constructed.

Objective To observe the effect of cognitive training combined with swallowing training real-time electrical stimulation on dysphagia and cognitive impairment after stroke. Methods 40 patients with dysphagia and cognitive impairment after stroke were randomly divided into observation group (n=20) and control group (n=20). The observation group accepted swallowing training (including routine vocal training and swallowing training real-time electrical stimulation) and cognitive training. The control group accepted routine vocal training and neuromuscular electrical stimulation. The cognitive function and swallowing function were accessed with Mini-Mental State Examination (MMSE) and Video Fluoroscopy Swallowing Study (VFSS) respectively before and 1 month after stroke. Results After treatment, the MMSE scores and the time for a bolus to pass the pharynx improved in both groups (P<0.05), and the observation group was superior to the control group (P<0.05). Conclusion Cognitive training combined with swallowing training real-time electrical stimulation can promote the recovery of swallowing function of patients with dysphagia and cognitive impairment after stroke.