HGF非融合蛋白真核表达载体的构建及其在骨骼肌细胞中的表达

目的构建以HGF为目的基因、以EGFP为报告基因的非融合蛋白真核表达载体pCMV-HGF-IRES-EGFP,并观察HGF基因在原代培养的大鼠骨骼肌细胞中的表达。方法利用BamHI单酶切质粒pcDNA3-HGF,得到HGF基因片段,将其正向插入到真核表达载体pCMV-IRES-EGFP的CMV后方BamHI的单克隆位点。脂质体介导转染至原代培养的大鼠骨骼肌细胞,在荧光显微镜下观察EGFP的表达,并用ELISA法检测HGF的表达情况。结果构建了HGF的非融合蛋白真核表达载体pCMV-HGF-IRES-EGFP,转染原代培养的大鼠骨骼肌细胞24~72h后,见30%的细胞表达EGFP。ELISA检测细胞培养液证实转染后第1天即有HGF的表达,第4天HGF浓度为(5402.0±227.9)ng/L。结论成功构建了非融合蛋白真核表达载体pCMV-HGF-IRES-EGFP,其在原代培养的大鼠骨骼肌细胞中能有效表达。     

低氧及运动致大鼠骨骼肌形态学改变的实验研究

目的观察低氧及低氧复合运动时,大鼠体重及腓肠肌显微、超微结构的变化,旨在探讨低氧状态下适当运动对骨骼肌的影响。方法建立大鼠低氧及低氧复合运动模型,运用光镜及透射电镜观察大鼠腓肠肌显微、超微结构的变化。结果低氧复合运动组较低氧安静组大鼠体重差异有显著性意义(P<0.05)。低氧复合运动组骨骼肌结构正常,线粒体数量增加,骨骼肌纤维增粗,毛细血管增生。结论低氧复合适当的运动能够维持骨骼肌的正常形态,增强骨骼肌的有氧代谢,起到保护骨骼肌的作用,从而提高骨骼肌的低氧适应能力,促进低氧习服。     

经皮手术治疗骶髂关节损伤的临床解剖学数字化研究

[目的]研究CT扫描三维重建技术对于骶髂关节骨折进行闭合手术治疗的术前指导作用。[方法]选择30个骶髂关节,均行X线片和螺旋CT三维重建后,随机分为3组,每组10个关节。A组:根据CT数据资料于M in ics软件指导下行骨折克氏针固定;B组:于电视X线机下行骨折克氏针固定,根据X线片和螺旋CT资料行骨折克氏针固定。固定后通过生物力学和大体剖面观察,评价不同组间生物力学和解剖学效果。[结果]A、B 2组在力学强度和解剖位置方面均优于C组,差异均有统计学意义(P<0.05),A组和B组在力学强度和解剖位置方面差异不显著(P>0.05)。[结论]Minics软件及三维重建技术可以很好的指导骶髂骨折的手术操作。     

Impaired modulation of motor cortex excitability by homonymous and heteronymous muscle afferents in focal hand dystonia.

A decrease of heteronymous median nerve-evoked inhibition of corticospinal projections to forearm extensor muscles was reported in a group of 10 dystonic patients by Bertolasi and colleagues in 2003. Here we tested the excitability of corticomotoneuronal connections to both wrist extensor (ECR) and flexor (FCR) muscles after conditioning stimulation of median and also radial nerve at rest in a group of 25 patients with focal hand dystonia compared to 20 healthy subjects. We also investigated the effect of the wrist dystonic posture, either in flexion or in extension, on the afferent modulation of ECR and FCR motor evolved potentials (MEPs). The heteronymous (median-induced) but also homonymous (radial-induced) inhibitions (interstimuli intervals 13-21 ms) of ECR MEP size observed in healthy subjects were decreased in patients. In addition, homonymous (median-induced) facilitation of FCR MEP size was also decreased in patients while heteronymous inhibition (radial-induced) was not. Neither the involvement of the target muscle in the dystonic posture nor the origin of the afferent volley (from a dystonic muscle) influenced the degree of impairment of afferent modulation of the MEP. These findings support the view that a global abnormal somatosensory coupling in focal hand dystonia may contribute to an inadequate motor command to wrist muscles.     

强化膨胀式椎弓根螺钉翻修作用的生物力学评价

目的测试骨水泥强化膨胀式椎弓根螺钉(augmentation expansive pedicle screwfixa-tion with polymethyl methacrylate,AEPS),未强化的膨胀式椎弓根螺钉(expansive pedicle screw,EPS),CDH椎弓根螺钉翻修后的最大轴向拔出力,评价AEPS螺钉的翻修效应。方法24个新鲜小牛腰椎随机分成三组,AEPS组、EPS组、CDH组,每组8个椎体,三组椎体的椎弓根处均预先植入直径6.5mm CDH螺钉,拔出螺钉造成螺钉松动脱出的脊柱模型,然后分别植入AEPS、EPS、CDH螺钉,再次拔出螺钉,记录翻修后最大轴向拔出力。结果AEPS、EPS、CDH螺钉翻修后的最大拔出力分别为(2723.3±565.4)、(2216.3±475.6)和(1321.4±346.7)N,AEPS和EPS螺钉最大拔出力均显著高于CDH螺钉(P<0.05),AEPS轴向拔出力高于EPS螺钉,但差异无统计学意义(P>0.05)。结论AEPS与EPS螺钉具有相当的翻修作用。     

Assessing the Stiffness of Spinal Fusion in Animal Models

The clinical goal of spinal fusion is to reduce motion and the associated pain. Therefore, measuring motion under loading is critical. The purpose of this study was to validate four-point bending as a means to mechanically evaluate simulated fusions in dog and rabbit spines. We hypothesized that this method would be more sensitive than manual palpation and would be able to distinguish unilateral vs bilateral fusion. Spines from four mixed breed dogs and four New Zealand white rabbits were used to simulate posterolateral fusion with polymethyl methacrylate as the fusion mass. We performed manual palpation and nondestructive mechanical testing in four-point bending in four planes of motion: flexion, extension, and right and left bending. This testing protocol was used for each specimen in three fusion modes: intact, unilateral, and bilateral fusion. Under manual palpation, all intact spines were rated as not fused, and all unilateral and bilateral simulated fusions were rated as fused. In four-point bending, dog spines were significantly stiffer after unilateral fusion compared with intact in all directions. Additionally, rabbit spines were stiffer in flexion and left bending after unilateral fusion. All specimens exhibited significant differences between intact and bilateral fusion except the rabbit in extension. For unilateral vs bilateral fusion, significant differences were present for right bending in the dog model and for flexion in the rabbit. Unilateral fusion can provide enough stability to constitute a fused grade by manual palpation but may not provide structural stiffness comparable to bilateral fusion.     

前列通瘀提取液对前列腺平滑肌细胞增殖与凋亡的作用

目的:研究前列通瘀提取液对体外培养前列腺基质平滑肌细胞增殖与凋亡的作用。方法:分别以1、2、10、50、100×10-5g/L 5种不同浓度前列通瘀提取液作用体外培养前列腺基质平滑肌细胞,采用MTT和TUNEL法分别测定抗增殖指数和凋亡指数。结果:5种不同浓度前列通瘀提取液作用48 h后抗增殖指数分别为50.61%、53.52%、56.92%、65.53%、72.94%,抗增殖效果随着浓度增大而增大。100×10-5g/L前列通瘀提取液作用24、48、72 h后凋亡指数与对照组比较差异无显著性。结论:(1~100)×10-5g/L前列通瘀提取液体外作用前列腺基质平滑肌细胞具有显著的抗增殖作用。     

Treatment of cremaster synkinesias with botulinum toxin A: a video case report.

Synkinesias secondary to nerve lesions and aberrant re-innervation are well-known phenomena especially after lesions of the facial nerve. Synkinesias can successfully be treated with botulinum toxin A (BTx A). Synkinesias of the cremaster muscle have not been described or treated to date. We present the case of a 62-year-old man who developed synkinesias of both cremaster muscles after extensive laparatomy for esophageal cancer. Treatment of synkinesias with various oral medications had been unsuccessful. Electromyography-guided injections of BTx A in both cremaster muscles (15 MU on the right and 10 on the left) led to significant symptom relief for an average of 8 weeks. We present the case including pre- and posttreatment video clips.     

103Pd放射性核素支架抑制损伤血管平滑肌细胞增殖的实验研究

[目的]采用兔腹主动脉损伤致再狭窄的动物模型,探讨钯-103(103Pd)放射性核素支架对损伤血管中膜平滑肌细胞增殖的影响。[方法]选用雄性新西兰兔50只,随机分为普通支架组及核素支架组,设正常对照。分别于术后3、7、14、28及56 d取材,进行病理形态学、透射电镜、免疫组化(增殖细胞核抗原PCNA、细胞周期素蛋白Cy-clin E)及原位杂交(C-myc mRNA)实验观察。[结果]①光镜下发现,核素支架组管腔狭窄程度明显低于普通支架组,术后第56天最显著(P<0.01);②透射电镜显示,术后3~28 d,核素支架组中膜血管平滑肌细胞增殖明显低于普通支架组,7d时达峰,为13.78％±0.64％vs19.53％±0.44％(P<0.01);③免疫组化显示,术后3~28 d核素支架组PCNA及Cyclin E表达均低于普通支架组,7 d为表达高峰,PCNA为16.35％±0.79％vs24.36±0.55％,Cy-clin E为14.78％±1.07％ vs 22.65％±1.00％(P<0.01)。④对c-myc mRNA进行原位杂交显示,术后3~28 d核素支架组的表达明显低于普通支架组,7 d达峰值,为17.48％±0.53％ vs 25.34％±0.87％(P<0.01);⑤C-mycmRNA与PCNA相关性分析提示二者呈明显正相关(P<0.01)。[结论]103Pd放射性核素支架通过抑制损伤血管中膜平滑肌细胞的增殖,降低再狭窄的程度,从而减少再狭窄的发生率。     

基于匹配实测表面肌电信号的模型参数辨识

目的 通过对仿真与真实表面肌电信号(sEMG)的波形匹配以及肌疲劳现象的分析,研究sEMG信号的模型参数辨识问题. 方法 在运动单位仿真的基础上,引入神经激励对运动单位的募集和发放控制特性,建立了一个较为完善的sEMG信号生理学模型.利用调整模型相关生理参数使仿真与真实sEMG信号的运动单位动作电位(MUAP)波形相匹配的方法,实现对模型参数进行估计,通过调节肌纤维传导速度(MFCV)使仿真与真实sEMG信号的平均频率(MNF)及中值频率(MDF)拟合直线趋势相似的方法,研究肌肉的疲劳现象及其机理. 结果 适当调节sEMG信号模型参数可使仿真信号波形逼近真实sEMG信号波形,各个肌纤维的MFCV在模拟恒力持续收缩过程中减小时,仿真信号的MNF和MDF拟合直线呈下降趋势. 结论 采用模型方法能够实现仿真与真实sEMG信号波形的良好匹配,并能够有效地表达肌肉的疲劳过程,可应用于肌电信号相关领域的研究.