目的为研究超大分子量肌小节蛋白肌联蛋白(titin)的生理病理功能,在一次电泳过程中同时分离titin各亚型和中分子量肌小节蛋白肌球蛋白重链(myosin heavy chain,MHC)。方法使用16cm×18cm垂直电泳系统,在电泳板下1/3灌注10g/L SDS-P...目的为研究超大分子量肌小节蛋白肌联蛋白(titin)的生理病理功能,在一次电泳过程中同时分离titin各亚型和中分子量肌小节蛋白肌球蛋白重链(myosin heavy chain,MHC)。方法使用16cm×18cm垂直电泳系统,在电泳板下1/3灌注10g/L SDS-PAGE胶,上2/3灌注60g/L SDS-琼脂糖(SDS-VAGE)胶。低温8℃下持续电泳5h,在电泳板上层以SDS-VAGE胶电泳分离titin亚型,下层以SDS-PAGE胶电泳分离MHC。电泳后VAGE胶使用银染法标记titin各亚型,PAGE胶使用考马斯亮蓝染色法标记MHC。结果 titin各亚型得到有效的分离,目标蛋白条带显示清晰,与其分子量大小一一对应,分离效果明确。结论一步法垂直电泳系统可应用于超大分子量蛋白的电泳,同时可分离多个分子量差距大的蛋白,提高蛋白电泳实验效率。展开更多
目的探究对重症肌无力合并胸腺病变的患者血清中乙酰胆碱受体(acetylcholine receptors,AchR)抗体、肌联蛋白(Titin)抗体和肌肉特异性酪氨酸激酶(muscle specific tyrosine kinase,MuSK)抗体的检测意义。方法选取我院收治的80例重症肌...目的探究对重症肌无力合并胸腺病变的患者血清中乙酰胆碱受体(acetylcholine receptors,AchR)抗体、肌联蛋白(Titin)抗体和肌肉特异性酪氨酸激酶(muscle specific tyrosine kinase,MuSK)抗体的检测意义。方法选取我院收治的80例重症肌无力患者,根据是否存在胸腺病变分为胸腺病变组48例和非胸腺病变组32例,另取80例健康个体作为对照组,通过免疫酶联吸附试验对3组成员血清中AchR抗体、Titin抗体和MuSK抗体进行检测并比较。结果重症肌无力患者血清AchR抗体和Titin抗体阳性率(76.3%,52.5%)均明显高于对照组(11.3%,5.0%)(P<0.05);重症肌无力组及对照组患者血清MuSK抗体阳性率均为0,但前者血清中MuSK抗体水平明显高于后者(P<0.05);AchR,Titin两种抗体联合检测可明显提高检测重症肌无力的灵敏度;AchR和Titin抗体在胸腺病变组中的阳性率明显高于非胸腺病变组(P<0.05);全身型重症肌无力患者的血清AchR抗体和Titin抗体阳性率明显高于眼肌型(P<0.05)。结论 AchR抗体和Titin抗体的阳性率与重症肌无力患者的病情相关,病情严重或合并胸腺病变可明显提高两者的阳性率,此外两者联合检测可提高诊断的灵敏度。而MuSK抗体在我国重症肌无力患者中的阳性率较低。展开更多
Objective: To determine the phenotype variability associated with the specific C-terminal M-line titin mutation known to cause autosomal dominant distal myo pathy, tibial muscular dystrophy (TMD; MIM 600334), and limb...Objective: To determine the phenotype variability associated with the specific C-terminal M-line titin mutation known to cause autosomal dominant distal myo pathy, tibial muscular dystrophy (TMD; MIM 600334), and limb girdle muscular dys trophy 2J (LGMD2J). Methods: Three hundred eighty-six individuals were genotype d for the Finnish founder mutation in titin (FINmaj) causing TMD/LGMD2J. Results : Two hundred seven patients were heterozygous for the mutation. Among these pat ients, 189 (91%) had a more common phenotype compatible with the classic descri ption of TMD. However, 18(9%) had unusual phenotypes such as proximal leg or po sterior lower leg muscle weakness and atrophy even at onset.Four patients were c onfirmed homozygotes representing the LGMD2J phenotype. These homozygotes were h alf of the eight LGMD patients previously described in the original large consan guineous kindred. Conclusions: Large variability of phenotypic expression caused by just one mutation, the Finnish FINmaj, suggests that no certain phenotype of myopathy/dystrophy can be excluded from being caused by mutated titin. Yet unkn own homozygous or compound heterozygous titin mutations without phenotype in the heterozygote carriers may be responsible for undetermined recessive MD and LGMD .展开更多
文摘Objective: To determine the phenotype variability associated with the specific C-terminal M-line titin mutation known to cause autosomal dominant distal myo pathy, tibial muscular dystrophy (TMD; MIM 600334), and limb girdle muscular dys trophy 2J (LGMD2J). Methods: Three hundred eighty-six individuals were genotype d for the Finnish founder mutation in titin (FINmaj) causing TMD/LGMD2J. Results : Two hundred seven patients were heterozygous for the mutation. Among these pat ients, 189 (91%) had a more common phenotype compatible with the classic descri ption of TMD. However, 18(9%) had unusual phenotypes such as proximal leg or po sterior lower leg muscle weakness and atrophy even at onset.Four patients were c onfirmed homozygotes representing the LGMD2J phenotype. These homozygotes were h alf of the eight LGMD patients previously described in the original large consan guineous kindred. Conclusions: Large variability of phenotypic expression caused by just one mutation, the Finnish FINmaj, suggests that no certain phenotype of myopathy/dystrophy can be excluded from being caused by mutated titin. Yet unkn own homozygous or compound heterozygous titin mutations without phenotype in the heterozygote carriers may be responsible for undetermined recessive MD and LGMD .
