The myosin heavy chain(MyHC)is one of the major structural and contracting proteins of muscle.We have isolated the cDNA clone encoding MyHC of the grass carp,Ctenopharyngodon idella. The sequence comprises 5 934 bp,in...The myosin heavy chain(MyHC)is one of the major structural and contracting proteins of muscle.We have isolated the cDNA clone encoding MyHC of the grass carp,Ctenopharyngodon idella. The sequence comprises 5 934 bp,including a 5 814 bp open reading frame encoding an amino acid sequence of 1 937 residues.The deduced amino acid sequence showed 69%homology to rabbit fast skeletal MyHC and 73%–76%homology to the MyHCs from the mandarin fish,walleye pollack,white croaker,chum salmon,and carp.The putative sequences of subfragment-1 and the light meromyosin region showed 61.4%–80%homology to the corresponding regions of other fish MyHCs.The tissue-specific and developmental stage-specific expressions of the MyHC gene were analyzed by quantitative real-time PCR.The MyHC gene showed the highest expression in the muscles compared with the kidney,spleen and intestine.Developmentally,there was a gradual increase in MyHC mRNA expression from the neural formation stage to the tail bud stage.The highest expression was detected in hatching larva.Our work on the MyHC gene from the grass carp has provided useful information for fish molecular biology and fish genomics.展开更多
Lung cancer is the leading cause of cancer related death in the United States killing over 130,000 people each year. While a combination of chemo and radiation therapy may be effective, surgery is still required for m...Lung cancer is the leading cause of cancer related death in the United States killing over 130,000 people each year. While a combination of chemo and radiation therapy may be effective, surgery is still required for many patients. Without surgery, the disease may progress and lead to metastases. We sought to determine if treatment with anti-non-muscle myosin IIA antibody would inhibit movement of the cells in the presence and absence of glabridin (an isoflavonoid compound shown to inhibit cell migration by inhibiting myosin). We compared inhibition by glabridin to that of an anti-non-muscle myosin IIA antibody and a combination therapy of both at 12 and 24 hours post wound creation. Cells that took up the anti-non-muscle myosin IIA antibody were greatly inhibited in motility and exhibited no significant change in wound healing. Glabridin treatment resulted in a dramatic increase in wound size within 12 hours and regeneration within 24 hours. The greatest decrease in motility was observed in cells treated with the combination of both glabridin and anti-non-muscle myosin IIA antibody. By 24 hrs, cell migration had halted due to death of the cells resulting from this combination. Further testing needs to be done to determine a safe mode of delivery of the combination therapy to ensure only local distribution. Controlled release drug delivery depot systems have been used as a means to provide local release of drugs intra-tumorally or adjacent to the cancerous tissue after surgical resection and have great potential.展开更多
The aim of this short review is to describe the role of myosin isoforms during the adaptation of skeletal muscle to prolonged physical activity (for example endurance exercise) and to show the coordination between cha...The aim of this short review is to describe the role of myosin isoforms during the adaptation of skeletal muscle to prolonged physical activity (for example endurance exercise) and to show the coordination between changes in muscle oxidative capacity and myofibrillar apparatus in slow-twitch and fast-twitch muscles. Adaptational changes in myosin isoforms during long lasting muscle activity (decrease of MyHC IIb isoforms relative content and increase of that MyHC IIa and decrease of MyLC 1 fast isoforms in fast-twitch muscles) are in good coordination with changes of muscle oxidative capacity. These changes show that during regular endurance exercise fast-twitch muscle fibers (type IIA) are also recruited and create the potential source of increase in endurance capacity during the process of adaptation to the prolonged physical activity.展开更多
Triple immunofluorescence staining has recently been developed to simultaneously identify all muscle fibers on a single cryosection which is helpful for clinical and basic research, but it has disadvantages such as fa...Triple immunofluorescence staining has recently been developed to simultaneously identify all muscle fibers on a single cryosection which is helpful for clinical and basic research, but it has disadvantages such as fast photobleaching and unclear outlines of muscle fibers. Triple immunoenzyme staining（TIE） is likely to avoid these disadvantages. In this study, we aimed to establish a sensitive and specific TIE technique to identify fiber types in normal, denervated, and reinnervated rat muscles, and to develop a systematic sampling method for muscle fiber quantification. Tibialis anterior and soleus from normal, denervated, and reinnervated Lewis rat hind limbs were used. Five consecutive cryosections were cut from each muscle, including one for TIE and four for single immunoenzyme staining（SIE）. The TIE was performed using the polymerized reporter enzyme staining system for the first two antigens（A4.74 for My HC-IIA, BA-F8 for My HC-I） and alkaline phosphatase staining system for the third antigen（BF-F3 for My HC-IIB）, followed by corresponding detective systems and respective chromogens. The type of muscle fibers was quantified by systematic sampling at 12.5%, 25%, 33% and 50% of all muscle fibers, and was compared with that acquired from counting all the fibers（100%）. All muscle fiber phenotypes, including pure and hybrid, could be simultaneously identified on a single TIE cryosection with clear outlines. The fiber types on TIE slides matched well with their respective counterpart on the consecutive SIE slides with a 95% match rate. Systematic sampling of 12.5% fibers could represent the true fiber type distribution of the entire muscle section. Our results suggest that novel TIE can effectively visualize fiber types in normal, denervated or reinnervated rat muscles.展开更多
目的:观察黄芪皂甙和丹参酮ⅡA对大鼠骨骼肌急性钝挫伤后胚胎型肌球蛋白重链(embryonic Myosin Heavy Chain,MHC-emb)和波形蛋白(vimentin)表达的影响。方法:80只雄性Wistar大鼠(体重180～200g)随机分为A组(黄芪皂甙组)、B组(丹参酮ⅡA...目的:观察黄芪皂甙和丹参酮ⅡA对大鼠骨骼肌急性钝挫伤后胚胎型肌球蛋白重链(embryonic Myosin Heavy Chain,MHC-emb)和波形蛋白(vimentin)表达的影响。方法:80只雄性Wistar大鼠(体重180～200g)随机分为A组(黄芪皂甙组)、B组(丹参酮ⅡA组)、C组(黄芪皂甙+丹参酮ⅡA组)、D组(黄芪丹参注射液组)、E组(生理盐水对照组),每组16只。采用打击装置造成右侧腓肠肌中段钝挫伤模型,分别于损伤局部注射相应药物,并于损伤后4、7、14和28天进行腓肠肌损伤部位取材。Western Blotting方法检测各组MHC-emb和波形蛋白表达量的变化。结果:①与生理盐水组相比,各干预组MHC-emb表达均明显升高,差异有统计学意义(P<0.05),这种情况一直持续到伤后28天;其中又以C组和D组升高最为明显;②与生理盐水组相比,各干预组波形蛋白表达均明显降低,差异有统计学意义(P<0.05),这种情况一直持续到伤后28天;其中又以C组和D组降低最为明显。结论:在急性骨骼肌钝挫伤修复过程中,黄芪皂甙、丹参酮ⅡA均能促进MHC-embmRNA表达,抑制波形蛋白mRNA表达,联合使用时效果最佳,与黄芪丹参复方成分基本一致。展开更多
目的研究低频复合生理频率慢性电刺激后肺气肿兔膈肌肌球蛋白重链(myosin heavy chain,MHC)亚型及其力学模式的适应性变化。方法采用木瓜蛋白酶雾化吸入法建立肺气肿模型:测定正常对照组、肺气肿组和肺气肿各慢性电刺激(chronic electri...目的研究低频复合生理频率慢性电刺激后肺气肿兔膈肌肌球蛋白重链(myosin heavy chain,MHC)亚型及其力学模式的适应性变化。方法采用木瓜蛋白酶雾化吸入法建立肺气肿模型:测定正常对照组、肺气肿组和肺气肿各慢性电刺激(chronic electrical stimulation,CES)组膈肌MHC亚型及膈肌颤搐收缩张力(Pt)、峰值张力时间(TPT)、1/2松弛时间(1/2Rt)、强直颤搐收缩张力(Po)、疲劳指数(FI)和疲劳恢复指数(FRI)的适应性变化。结果①40Hz、10Hz、(10+40)Hz慢性电刺激后肺气肿兔膈肌MHC的相对含量较刺激前都明显增加(P<0·05)。10Hz组MHC-I型比例明显增加(P<0·01),(10+40)Hz组MHC-I、MHC-IIa型比例增加(P<0·05)。②肺气肿组Pt、Po均较正常对照组明显减低(P<0·01),TPT、1/2Rt延长(P<0·01),FI、FRI增加(P<0·01);(10+40)Hz组和40Hz组与肺气肿组比较Pt、Po均明显增加(P<0·01),TPT、1/2Rt均明显缩短(P<0·01),FI和FRI下降(P<0·01);(10+40)Hz组与40Hz组比较,Pt、Po、TPT、1/2Rt无显著性差异(P>0·05),FI和FRI降低(P<0·01)。结论不同频率的CES可导致膈肌MHC亚型及肌条力学产生不同的适应性变化,低频复合生理频率电刺激可显著提高肺气肿兔膈肌收缩力,增强膈肌抗疲劳能力,可能是体外膈肌起搏对COPD患者膈肌康复治疗更佳的频率模式之一。展开更多
基金Supported by the National Natural Science Foundation of China(Nos.30972263,30771644)the Natural Science Foundation of HunanProvince(No.09jj6037)
文摘The myosin heavy chain(MyHC)is one of the major structural and contracting proteins of muscle.We have isolated the cDNA clone encoding MyHC of the grass carp,Ctenopharyngodon idella. The sequence comprises 5 934 bp,including a 5 814 bp open reading frame encoding an amino acid sequence of 1 937 residues.The deduced amino acid sequence showed 69%homology to rabbit fast skeletal MyHC and 73%–76%homology to the MyHCs from the mandarin fish,walleye pollack,white croaker,chum salmon,and carp.The putative sequences of subfragment-1 and the light meromyosin region showed 61.4%–80%homology to the corresponding regions of other fish MyHCs.The tissue-specific and developmental stage-specific expressions of the MyHC gene were analyzed by quantitative real-time PCR.The MyHC gene showed the highest expression in the muscles compared with the kidney,spleen and intestine.Developmentally,there was a gradual increase in MyHC mRNA expression from the neural formation stage to the tail bud stage.The highest expression was detected in hatching larva.Our work on the MyHC gene from the grass carp has provided useful information for fish molecular biology and fish genomics.
文摘Lung cancer is the leading cause of cancer related death in the United States killing over 130,000 people each year. While a combination of chemo and radiation therapy may be effective, surgery is still required for many patients. Without surgery, the disease may progress and lead to metastases. We sought to determine if treatment with anti-non-muscle myosin IIA antibody would inhibit movement of the cells in the presence and absence of glabridin (an isoflavonoid compound shown to inhibit cell migration by inhibiting myosin). We compared inhibition by glabridin to that of an anti-non-muscle myosin IIA antibody and a combination therapy of both at 12 and 24 hours post wound creation. Cells that took up the anti-non-muscle myosin IIA antibody were greatly inhibited in motility and exhibited no significant change in wound healing. Glabridin treatment resulted in a dramatic increase in wound size within 12 hours and regeneration within 24 hours. The greatest decrease in motility was observed in cells treated with the combination of both glabridin and anti-non-muscle myosin IIA antibody. By 24 hrs, cell migration had halted due to death of the cells resulting from this combination. Further testing needs to be done to determine a safe mode of delivery of the combination therapy to ensure only local distribution. Controlled release drug delivery depot systems have been used as a means to provide local release of drugs intra-tumorally or adjacent to the cancerous tissue after surgical resection and have great potential.
文摘The aim of this short review is to describe the role of myosin isoforms during the adaptation of skeletal muscle to prolonged physical activity (for example endurance exercise) and to show the coordination between changes in muscle oxidative capacity and myofibrillar apparatus in slow-twitch and fast-twitch muscles. Adaptational changes in myosin isoforms during long lasting muscle activity (decrease of MyHC IIb isoforms relative content and increase of that MyHC IIa and decrease of MyLC 1 fast isoforms in fast-twitch muscles) are in good coordination with changes of muscle oxidative capacity. These changes show that during regular endurance exercise fast-twitch muscle fibers (type IIA) are also recruited and create the potential source of increase in endurance capacity during the process of adaptation to the prolonged physical activity.
基金sponsored by the Armed Forces Institute of Regenerative Medicine award number W81XWH-08-2-0034The U.S.Army Medical Research Acquisition Activity,820 Chandler Street,Fort Detrick MD 21702-5014 is the awarding and administering acquisition office+4 种基金The content of the manuscript does not necessarily reflect the position or the policy of the Government,and no official endorsement should be inferredsupported(AJW)by CTSA Grants Number UL1 TR000135 and TL1 TR000137 from the National Center for Advancing Translational Science(NCATS)supported by the Sundt Fellowship fund,Department of Neurological Surgery,Mayo Clinicsupported by the Helene Houle Career Development Award in Neurologic Surgery ResearchFund for the Center for Regenerative Medicine Program Director,Neuroregenerative Medicine,Mayo Clinic
文摘Triple immunofluorescence staining has recently been developed to simultaneously identify all muscle fibers on a single cryosection which is helpful for clinical and basic research, but it has disadvantages such as fast photobleaching and unclear outlines of muscle fibers. Triple immunoenzyme staining（TIE） is likely to avoid these disadvantages. In this study, we aimed to establish a sensitive and specific TIE technique to identify fiber types in normal, denervated, and reinnervated rat muscles, and to develop a systematic sampling method for muscle fiber quantification. Tibialis anterior and soleus from normal, denervated, and reinnervated Lewis rat hind limbs were used. Five consecutive cryosections were cut from each muscle, including one for TIE and four for single immunoenzyme staining（SIE）. The TIE was performed using the polymerized reporter enzyme staining system for the first two antigens（A4.74 for My HC-IIA, BA-F8 for My HC-I） and alkaline phosphatase staining system for the third antigen（BF-F3 for My HC-IIB）, followed by corresponding detective systems and respective chromogens. The type of muscle fibers was quantified by systematic sampling at 12.5%, 25%, 33% and 50% of all muscle fibers, and was compared with that acquired from counting all the fibers（100%）. All muscle fiber phenotypes, including pure and hybrid, could be simultaneously identified on a single TIE cryosection with clear outlines. The fiber types on TIE slides matched well with their respective counterpart on the consecutive SIE slides with a 95% match rate. Systematic sampling of 12.5% fibers could represent the true fiber type distribution of the entire muscle section. Our results suggest that novel TIE can effectively visualize fiber types in normal, denervated or reinnervated rat muscles.
文摘目的:观察黄芪皂甙和丹参酮ⅡA对大鼠骨骼肌急性钝挫伤后胚胎型肌球蛋白重链(embryonic Myosin Heavy Chain,MHC-emb)和波形蛋白(vimentin)表达的影响。方法:80只雄性Wistar大鼠(体重180～200g)随机分为A组(黄芪皂甙组)、B组(丹参酮ⅡA组)、C组(黄芪皂甙+丹参酮ⅡA组)、D组(黄芪丹参注射液组)、E组(生理盐水对照组),每组16只。采用打击装置造成右侧腓肠肌中段钝挫伤模型,分别于损伤局部注射相应药物,并于损伤后4、7、14和28天进行腓肠肌损伤部位取材。Western Blotting方法检测各组MHC-emb和波形蛋白表达量的变化。结果:①与生理盐水组相比,各干预组MHC-emb表达均明显升高,差异有统计学意义(P<0.05),这种情况一直持续到伤后28天;其中又以C组和D组升高最为明显;②与生理盐水组相比,各干预组波形蛋白表达均明显降低,差异有统计学意义(P<0.05),这种情况一直持续到伤后28天;其中又以C组和D组降低最为明显。结论:在急性骨骼肌钝挫伤修复过程中,黄芪皂甙、丹参酮ⅡA均能促进MHC-embmRNA表达,抑制波形蛋白mRNA表达,联合使用时效果最佳,与黄芪丹参复方成分基本一致。
文摘目的研究低频复合生理频率慢性电刺激后肺气肿兔膈肌肌球蛋白重链(myosin heavy chain,MHC)亚型及其力学模式的适应性变化。方法采用木瓜蛋白酶雾化吸入法建立肺气肿模型:测定正常对照组、肺气肿组和肺气肿各慢性电刺激(chronic electrical stimulation,CES)组膈肌MHC亚型及膈肌颤搐收缩张力(Pt)、峰值张力时间(TPT)、1/2松弛时间(1/2Rt)、强直颤搐收缩张力(Po)、疲劳指数(FI)和疲劳恢复指数(FRI)的适应性变化。结果①40Hz、10Hz、(10+40)Hz慢性电刺激后肺气肿兔膈肌MHC的相对含量较刺激前都明显增加(P<0·05)。10Hz组MHC-I型比例明显增加(P<0·01),(10+40)Hz组MHC-I、MHC-IIa型比例增加(P<0·05)。②肺气肿组Pt、Po均较正常对照组明显减低(P<0·01),TPT、1/2Rt延长(P<0·01),FI、FRI增加(P<0·01);(10+40)Hz组和40Hz组与肺气肿组比较Pt、Po均明显增加(P<0·01),TPT、1/2Rt均明显缩短(P<0·01),FI和FRI下降(P<0·01);(10+40)Hz组与40Hz组比较,Pt、Po、TPT、1/2Rt无显著性差异(P>0·05),FI和FRI降低(P<0·01)。结论不同频率的CES可导致膈肌MHC亚型及肌条力学产生不同的适应性变化,低频复合生理频率电刺激可显著提高肺气肿兔膈肌收缩力,增强膈肌抗疲劳能力,可能是体外膈肌起搏对COPD患者膈肌康复治疗更佳的频率模式之一。
