ABSTRACT
Objective To construct a gene recombinant lentiviral vector pCMV -G -U6 -hHGF and detect its expression in C2C12 myoblast cells .Methods hHGF gene fragments were obtained and purified by RT -PCR method ,and were cloned to pCMV -G&NR -U6 ,then the restructured lentiviral vector was transformed into e . coli DH5 alpha ,the positive colonies were identified by BamHI and Hind Ⅲ enzyme digestion .The selected positive colonies were tested by PCR and sequencing analysis .The expression plasmids and packing plasmids were co -trans_fected into 293 T cells ,and virus titer was observed under the fluorescence microscope .Furthermore ,transfected C2C12 cells with lenti virus ,and the expression of HGF was detected by PCR and WB methods .ResuIts PCR and sequencing analysis showed that the lentiviral vector was constructed correctly and successfully ,the virus titer was above 1 x 109 IU/mL .The results of PCR and WB showed that HGF expression level in the lentiviral vector group was much higher than those of in blank control and negative control groups ,and yet the expression was stable after 72 hours .ConcIusion The lentiviral vector pCMV -G﹠NR -U6 -hHGF has been successfully constructed ,and stable expressed in C2C12 cells .It provides references for experimental study in the fields of the denervated skeletal muscle fibrosis and nerve regeneration treatment .
ABSTRACT
BACKGROUND AND OBJECTIVES : Several studies have reported that (18)F-fluorodeoxyglucose (FDG) uptake in positron emission tomography (PET) imaging is physiologically increased at the intact vocal cord in patients with unilateral vocal cord paralysis, which is explained by a compensatory mechanism of the intact vocal cord. We aimed to evaluate internal laryngeal muscles related to phonation and the compensatory mechanism in patients with unilateral vocal cord paralysis. SUBJECTS AND METHOD : We performed (18)FDG-PET imaging and neck computed tomography (CT) scan in the normal control group composed of 13 subjects and the paralyzed group composed of 11 patients with unilateral vocal cord paralysis. The two groups were divided into two groups, phonating and silent, before performing (18)FDG-PET. (18)FDG-PET and neck CT images by Syntegra. A specialist in nuclear medicine performed all the test measurements, the standardized uptake value (SUV) in the interarytenoid muscle (IA), both thyroarytenoid muscles (TA), and both lateral cricoarytenoid muscles (LCA). The mean SUVs were statistically analyzed. RESULTS : In the Normal-Phonating group, the mean SUV of IA was the highest, with 3.68+/-0.96 (Mean+/-SD), followed by that of LCA, with 2.34+/-0.67. However, when compared with the same muscles in the Phonating-Silent group, only the SUV of IA was significantly increased by phonation. In the Paralyzed-Silent group, the SUV of TA in the intact side was the highest, with 2.30+/-0.39. In the Paralyzed-Phonating group, the SUV of TA in the intact side, IA, and LCA in the intact side were 5.88+/-2.65, 3.92+/-1.65, and 3.87+/-1.37, respectively. When compared with the same muscles in the Phonating-Silent group, the SUVs of TA and IA were significantly increased. CONCLUSION : The muscle related to the compensatory mechanism in patients with unilateral vocal cord paralysis is thyroarytenoid muscle in the intact side. The interarytenoid muscle plays a major role in the mechanism of phonation in humans.
Subject(s)
Humans , Laryngeal Muscles , Muscles , Neck , Nuclear Medicine , Phonation , Positron-Emission Tomography , Specialization , Vocal Cord Paralysis , Vocal CordsABSTRACT
The synchronous electromyography of the extrinsic laryngeal muscles (sternohyoid muscle, sternothyroid muscle and thyrohyoid muscle) and thyroarytenoid muscle of dogs was analyzed in this study. The result showed that the electrical activity of the thyrohyoid muscle were synchronous to that of the thyroaryienoid muscle during quiet respiration, deep inspiration, phonation and swallowing. The discharge of the sternohyoid muscle was recorded during expiration and phonation. The electrical activity of the sternothyroid muscle was also recorded during inspiration and phonation, and was increased during deep inspiration. The study suggests that reinnervation of the thyroarytenoid muscle from thyrohyoid branch of the ansa cervicalis for functional rehabilitation could fit well the physiological characteristic of the larynx.