复方中药与延迟性肌肉损伤血清学指标的变化：随机对照实验

背景：目前有关复方中药用于运动延迟性肌肉损伤的研究较少，仅有的几项针对单味中药成分对肌肉损伤的治疗效果的研究。 目的：通过复方中药和云南白药、阴性对照组大鼠在大强度训练后血清指标变化的对比分析，来反映复方中药对抗延迟性肌肉损伤的效果。 设计、时间及地点：随机对照动物实验，于2005-06/2006-01在广西师范大学体育学院生理实验室完成。 材料：8周龄纯系雄性昆明种大鼠54只，体质量200~220 g。复方中药的主要有效成分由秦归、白术、白芍、白芷和麝香等组成；云南白药为云南白药集团生产。 方法：根据大鼠的体质量随机将大鼠分为复方药物安静、即刻和恢复组，云南白药安静、即刻和恢复组，空白对照安静、即刻和恢复组，共9组，除了正常饮食外，复方药物组和云南白药组大鼠分别灌喂复方中药和云南白药0.78 g/(kg•d)，1次/d，给药4周，对照组灌喂相等量的生理盐水(0.5 mL)，即刻组和恢复组大鼠进行定量负荷游泳运动，1次/d，连续4周；安静组不运动。给药最后1 d，所有大鼠进行延迟性肌肉损伤运动。 主要观察指标：即刻组大鼠在运动后即取血样，恢复组在恢复24 h后，取血样，安静组随时间安排取血样，检测各组大鼠肌酸激酶、乳酸脱氢酶、血尿素氮变化。 结果：发现复方中药组大鼠在运动训练至力竭后大鼠血清肌酸激酶和乳酸脱氢酶活性的变化较其他组较大，血尿素氮上升的幅度较低，与对照组相比差异有显著性意义(P < 0.05)。 结论：复方中药对防治延迟性肌肉损伤具有较为明显的效果。     

实验大鼠采血方法的选择

采用300只SD大鼠进行采血。腹主动脉采血法在腹主动脉分叉处向心端1~ 3 mm处为最佳穿刺点，成功率为93.6%。眼眶后静脉采血法将针垂直插入内眦并向眼底方向转动以便切开静脉丛，成功率为89.9%。心脏穿刺采血法于剑突下以25°~30°斜行向上进针刺入皮下，针尖穿过横膈膜继续斜行刺入2.5~3.0 cm，成功率为83.4％。尾尖采血法以手术剪剪去尾尖5~10 mm，成功率为94.4%。颈静脉法于第4根肋骨水平方向刺入皮肤，向颈静脉进针，与胸部表面成30°~40°向颈静脉刺入，刺入的深度约5 mm，成功率为80.9%。腹主动脉采血法适用于取血量大的实验，不易溶血，不损伤器官，不会出现因操作不当造成的气栓与瘀血，有利于病理组织学检查。几种方法各有优缺点及注意事项，应根据不同的实验设计和要求来选择采血方式。     

长期运动预适应模型大鼠心脏蛋白激酶C和热休克蛋白70的表达

背景：研究表明蛋白激酶C和热休克蛋白70可能参与运动预适应的心脏保护作用。 目的：观察长期运动预适应对大鼠心脏蛋白激酶C和热休克蛋白70的影响及其对心脏保护的作用机制。 方法：将SD大鼠随机分为对照组，力竭运动组和运动预适应组。对照组和力竭运动组大鼠常规饲养3周，运动预适应组大鼠进行3周的间歇性游泳运动建立长期运动预适应动物模型。3周后，力竭运动组和运动预适应组大鼠进行一次性力竭游泳运动。 结果与结论：大鼠力竭运动后，力竭运动组心脏热休克蛋白70表达高于对照组(P < 0.05)；先经3周运动预适应再进行力竭运动后，运动预适应组心脏蛋白激酶C和热休克蛋白70的表达高于力竭运动组(P < 0.05)。结果证实，长期运动预适应能激活心脏蛋白激酶C，诱导热休克蛋白70的合成增多，从而发挥其心脏保护作用。     

无氧间歇训练对大鼠骨骼肌、心肌、肝脏自由基代谢的影响

背景：无氧间歇训练法是一种被广泛应用的重要训练方法，有关此训练对机体自由基代谢及其抗氧化酶活性影响的系统性研究较少。 目的：观察无氧间歇训练对大鼠体内自由基代谢的影响。 设计、时间及地点：随机对照动物实验，于2008-03/05在徐州师范大学运动生理实验室完成。 材料：健康雄性SD大鼠32只，体质量230~270 g，随机分为安静对照组，间歇训练对照组，间歇运动即刻组，间歇训练即刻组，每组8只。 方法：安静对照组不进行训练，间歇训练对照组和间歇训练即刻组进行跑步运动，以坡度10°，速度26.8 m/min运动1 min，后以坡度为0，速度5 m/min休息3 min，每天连续20次不间断，5 d/周，运动6周。间歇运动即刻组只在第6周最后一天进行一次运动。 主要观察指标：6周运动结束后即刻麻醉下处死各组大鼠，测定其骨骼肌，心肌，肝脏超氧化物歧化化酶，谷胱甘肽过氧化物酶，过氧化氢酶，丙二醛。 结果：①训练组运动后即刻各组织超氧化物歧化酶活性均显著高于间歇运动即刻组(P < 0.05)。②间歇运动即刻组股四头肌、心肌谷胱甘肽过氧化物酶活性显著低于安静对照组(P < 0.05)，丙二醛含量显著高于安静对照组(P < 0.05)，股四头肌和肝脏过氧化氢酶活性显著高于安静对照组(P < 0.05)。③间歇训练即刻组股四头肌、心肌谷胱甘肽过氧化物酶活性显著高于间歇运动即刻组(P < 0.05)，丙二醛含量明显低于间歇运动即刻组(P < 0.05)，股四头肌和心肌过氧化氢酶活性显著高于间歇运动即刻组(P < 0.05)。 结论：无氧间歇训练可以使机体抗氧化酶活性发生适应性变化，自由基清除能力增强。     

茶多酚对力竭运动大鼠骨骼肌组织氧化损伤的保护作用

背景：骨骼肌在急性运动时产生大量自由基,使体内氧化剂与抗氧化剂之间平衡的失调,引起骨骼肌疲劳、肌肉功能障碍和损伤等,使机体运动能力下降。而茶多酚是从茶叶中提取的多酚类物质,具有很强的抗氧化作用,是一种高效低毒的天然抗氧化剂。 目的：通过茶多酚对骨骼肌运动性损伤的干预,观察其对骨骼肌氧化损伤的保护作用。 方法：雄性SD健康大鼠30只,随机分为对照组、运动组、茶多酚组。茶多酚组大鼠补充茶多酚1周后,与运动组大鼠一起通过一次性力竭游泳建立力竭运动模型,记录大鼠力竭时间,测定力竭运动即刻时大鼠骨四头肌中丙二醛,超氧化物歧化酶谷胱甘肽过氧化物酶和总抗氧化能力。 结果与结论：力竭运动使大鼠骨骼肌丙二醛含量增加,超氧化物歧化酶、谷胱甘肽过氧化酶、血浆总抗氧化能力活力下降。茶多酚可以降低运动后丙二醛水平,提高运动后大鼠骨骼肌超氧化物歧化酶、谷胱甘肽过氧化酶、血浆总抗氧化能力活力,延长大鼠力竭时间,由此得出,大鼠力竭运动后骨骼肌组织会发生氧化损伤,茶多酚干预可能会保护骨骼肌氧化损伤,进而提高大鼠运动能力。     

不同强度力竭运动模型大鼠肝细胞凋亡及相关变化

背景：国内外不少实验证明,不同运动方式容易造成肝损伤,导致不同程度的肝细胞凋亡,其具体机制尚不明确。 目的：建立不同强度力竭运动模型,观察运动后大鼠肝细胞凋亡和肝糖元、NO、钙浓度变化。 设计、时间及地点：随机对照动物实验,超微结构观察,于2004-01/2006-12在湖南师范大学体育学院运动人体科学实验室、中南大学组胚实验室完成。 材料：30只8周龄雄性SD大鼠,体质量(219.2±19.5) g,根据Berdford运动模型将大鼠以随机数字表法分为对照组,中等强度、大强度力竭运动组,每组10只。 方法：运动组先进行3 d的适应性跑台训练,速度为10 m/min,坡度为0°。休息3 d后,中等强度力竭运动组初始速度为10 m/min,持续12 min,逐渐增加运动负荷,达到速度为19.3 m/min,持续到力竭。大强度力竭运动组初始速度为26.8 m/min,持续到力竭。共30 d,1次/d。对照组不进行运动训练。 主要观察指标：运动后即刻取肝组织检测肝糖元、NO和Ca2+及肝细胞凋亡情况。 结果：30只大鼠全部进入结果分析。不同强度力竭运动组大鼠都完成了运动,整个运动过程中未出现拒跑现象,中等强度力竭运动组力竭运动时间为(234.60±60.05) min,大强度力竭运动组力竭运动时间为(92.40±34.61) min。与对照组比较,两种强度力竭运动后,大鼠肝组织肝糖元含量、NO浓度均下降,线粒体Ca2+浓度、肝细胞凋亡指数均升高(P < 0.05,P < 0.01),中等强度力竭运动组效果更明显(P < 0.05)。 结论：中、大强度力竭运动均可导致大鼠肝细胞凋亡,肝糖元含量、NO浓度下降,线粒体Ca2+浓度升高,中等强度力竭运动效果更为显著,可能与力竭运动时间长有关。     

离心运动对大鼠骨骼肌细胞增殖和波形蛋白表达的时序性影响

背景：骨骼肌损伤后是通过肌卫星细胞的增殖形成新核来生长和修复的,但关于骨骼肌细胞增殖和波形蛋白表达的关系很少报道。 目的：探讨骨骼肌细胞增殖与波形蛋白表达的关系以及运动性骨骼肌微损伤后修复的机制。 设计、时间和地点：随机对照动物实验,于2007-12/2008-09在湖南师大运动人体科学实验中心完成。 材料：健康成年8周龄雄性SD大鼠50只,随机分成对照组和运动后即刻、运动后3 h、运动后24 h和运动后48 h组,每组10只大鼠。 方法：运动组大鼠进行重复3 d的力竭性离心运动,力竭模型采用跑台运动,速度为16 m/min,坡度为-16°,持续运动至力竭,对照组为正常大鼠,未做运动。 主要观察指标：运动组分别于运动后即刻,3,24,48 h取材,对照组一次性取材,免疫组织化学法检测各组大鼠肱三头肌内侧头不同恢复时相增殖细胞核抗原PCNA的表达和波形蛋白的表达。 结果：骨骼肌细胞增殖出现时序性变化,运动后即刻增殖指数显著大于对照组,运动后24 h达到峰值,运动后48 h增殖指数有所下降。骨骼肌细胞中波形蛋白表达出现时序性,而且其免疫反应分值的时序性与增殖指数出现一致性,但与增殖指数不具有相关性。 结论：3 d重复性力竭离心运动后骨骼肌细胞增殖和波形蛋白的表达出现时序性变化。波形蛋白的表达与肌细胞增殖具有一定的关系,但不是惟一的影响的因素。     

骨骼肌细胞线粒体通透性转换孔及bcl-2和bax mRNA表达与运动

背景：运动影响骨骼肌细胞的凋亡,而线粒体途径是介导细胞凋亡的一个重要途径。 目的：研究运动对大鼠骨骼肌线粒体通透性转换孔、凋亡调控基因bcl-2和bax表达的影响。 方法：将24只成年雄性SD大鼠随机分为3组：对照组正常饲养,6周游泳训练组进行6周的游泳训练,每周6次,一次性游泳力竭组于第6周进行一次力竭性游泳运动。应用紫外分光光度仪检测各组大鼠骨骼肌线粒体通透性转换孔的开放情况,应用RT-PCR测定大鼠骨骼肌bcl-2和bax mRNA的表达。 结果与结论：与对照组比较,6周游泳训练组大鼠骨骼肌线粒体通透性转换孔的开放程度变化不明显,bcl-2 mRNA的表达显著增加,bax mRNA的表达显著减少,bcl-2/bax mRNA比值显著增大(P < 0.01)。与对照组比较,一次性游泳力竭组大鼠骨骼肌线粒体通透性转换孔开放程度明显增加(P < 0.01),bcl-2 mRNA的表达显著减少,bax mRNA的表达显著增加,bcl-2/bax mRNA比值显著减小(P < 0.01)。说明运动训练可通过改变线粒体通透性转换孔的开放、调节bcl-2/bax表达,调控骨骼肌细胞凋亡。     

间歇性低氧训练复合运动训练模型大鼠的造血功能

背景：已有的研究显示低氧训练能够通过改善血液学指标提高有氧运动能力,但对于不同间歇低氧训练模式对血液学指标影响的研究较少。 目的：研究间歇性低氧训练对大鼠造血功能的影响。 方法：将雄性SD大鼠96只随机分为4组：低氧组和运动组分别进行间歇性低氧训练和游泳训练;低氧+运动组间歇性低氧训练和游泳训练同时进行;对照组自由活动和摄食。各组进行正式训练4周,分别于正式训练1,2,3,4周训练结束后24 h,取尾静脉血,检测大鼠血红蛋白、红细胞、及红细胞压积水平。 结果与结论：经过4周训练后,3种干预模式对大鼠血常规指标都有不同程度的影响,但低氧+运动组大鼠血红蛋白、红细胞、及红细胞压积指标改善最为显著。提示间歇性低氧训练复合运动训练在改善大鼠造血功能方面优于单独间歇性低氧训练或运动训练。     

微量全血单细胞凝胶电泳评定过度训练大鼠淋巴细胞DNA损伤

背景：微量全血单细胞凝胶电泳技术是检测有核细胞DNA损伤与修复的一种方法，具有敏感性高、取材方便、细胞制备简单、不需体外活化、费用低、实验周期短等优点。 目的：采用微量全血单细胞凝胶电泳技术评定过度训练对大鼠血液淋巴细胞DNA损伤的情况。 设计、时间及地点：随机分组设计、动物对照实验，于2004-08/09在沈阳体育学院研究生实验室完成。 材料：雄性Wistar大鼠29只，按随机数字表法分为2组，正常对照组12只、运动组17只。 方法：正常对照组大鼠不参加运动。运动组大鼠在玻璃池中进行游泳训练。采用6周递增负荷游泳训练建立过度训练动物模型，训练结束后24 h，避光眼眶取血。 主要观察指标：采用微量全血单细胞凝胶电泳法检测单个血液淋巴细胞的DNA尾长、尾矩、椭圆矩，以反映DNA的损伤情况。 结果：荧光显微镜下运动组大鼠淋巴细胞的DNA断裂比正常对照组明显，断片离开头部向阳极方向迁移，形成一个像彗星样的拖尾。运动组反映大鼠血液淋巴细胞DNA损伤的指标尾长、尾矩和椭圆矩显著高于正常对照组，差异有显著性意义 (P < 0.01）。 结论：6周递增负荷过度训练后大鼠血液淋巴细胞存在DNA损伤。     

大鼠肾移植技术改进与分析

背景：大鼠肾移植模型是器官移植和肾脏相关疾病研究中常用的动物模型,目前已有多种建模方式,但这些模型在移植时间、移植效果等方面都存在各种问题。 目的：拟建立一种简便、安全、成功率高的单侧供肾大鼠肾移植模型。 设计、时间及地点：动物模型分析实验,于2008-03/06在复旦大学泌尿外科研究所完成。 材料：Wistar大鼠作供体,SD大鼠作为受体,各60只。 方法：切取供体左侧肾脏作为供肾,分别以供体腹主动脉、左肾静脉与受体腹主动脉、下腔静脉端侧吻合,供肾输尿管以5-0带针线牵引插入膀胱,吻合缝针破口完成尿路重建。肾包膜下切除受体原有双肾。统计移植时间和移植成功率。 主要观察指标：移植时间、热缺血时间、冷缺血时间、动静脉吻合和尿路重建时间、移植失败率及原因、受体大鼠存活率。 结果：手术时间100~140 min。其中供肾热缺血时间8~10 s,冷缺血时间40~60 min。移植成功率为85%,失败原因分别为吻合口狭窄伴血栓6.67%、吻合口出血5%,感染1.67%,不明原因1.67%。 结论：实验建立的建模方法简便、可靠,明显缩短热缺血及尿路重建的时间,减少相关并发症。     

Cerebrospinal fluid sampling by lumbar puncture in rats - repeated measurements of nitric oxide metabolites

The ability to measure nitric oxide (NO) metabolites (NO2- plus NO3-: NOx) in cerebrospinal fluid (CSF) will facilitate understanding the involvement of NO in neurogenic or inflammatory diseases. The purposes of this study were to develop a reliable method for CSF sampling from the lumbar region, and to repeatedly measure NOx in naive rats. NOx in CSF were measured using the Griess method. Twelve young (13-week-old) and seven middle-aged (40-week-old) male Wistar rats were used. CSF (50-70 microl) was collected four times at 1-week intervals. The success rate of CSF collection was 96% and average surgery time was 21 min. The blood contamination rate was 11% on macroscopic inspection. NOx in the CSF ranged from 3.8 to 10.6 microM. The NOx in clear CSF were not significantly different from those with blood contamination on macroscopic inspection. There was, however, a linear correlation between the increase in NOx and the volume of venous blood added experimentally. NOx levels were significantly higher (P < 0.05) in young rats (6.5+/-0.2 microM) than in middle-aged rats (5.6+/-0.3 microM). There was no significant difference in CSF NOx among the four samples collected at 1-week intervals in 13-week-old rats. These results indicate that our CSF sampling technique can be used to reliably obtain a small amount of CSF for NOx measurement. This technique will facilitate further experimental studies of the involvement of CSF NO in neurogenic or inflammatory diseases.     

Rheologic changes in venous blood during prolonged sitting

We examined blood rheologic changes, mainly blood viscosity, during long-term quiet sitting. Samples of blood were obtained from the arm and foot veins of the healthy adult male volunteers before and after 2 h of quiet sitting. Blood viscosity, was immediately determined with an oscillation viscometer, and blood count and blood chemistry were analyzed. We found that the viscosity in blood from a foot vein but not from an arm vein increased significantly after 2 h of sitting. Our results show that 2 h of quiet sitting can increase thrombotic tendency locally in the leg but not systemically and suggests the importance of measuring focal venous blood viscosity.     

Experimental intracerebral haemorrhage: description of a semi-coagulated autologous blood model in rats

Abstract

Objectives:

A major limitation of intracerebral haemorrhage (ICH) research is the lack of reproducible animal models. The purpose of the present study was to produce experimental haemorrhages and to characterise the lesion by histology and behaviour in rats.

Methods:

A total of 180 male SD rats were anaesthetised with chloral hydrate. Rats were placed in a stereotactic frame, and a microinjector was introduced through a burr hole into the right striatum. Each rat received an injection of 60?μL of semi-coagulated autologous whole blood over 6?minutes. The needle was slowly withdrawn 40?minutes after the injection. Control rat had only needle insertion. Time courses of haematoma resolution and pathological changes after intrastriatal injection of semi-coagulated autologous blood were observed. Neurological status was evaluated on days 1, 3, 5, 7, 14 and 21.The behavioural tests used were forelimb placing and a corner turn test.

Results:

Rats with ICH had a marked, persistent neurological deficit and a highly reproduced haematoma in shape and size. Histologically, haematoma was observed at 1, 3, 5 and 7?days and cysts at 3?weeks. Behavioural abnormalities were present for 14?days, with the recovery of function occurring during the third week.

Conclusions:

The present ICH model in rats produces a consistent neurological deficit and reproducible haematoma in shape and size. This model could be useful to evaluate the future pharmaceutical therapies in ICH.     

DNA brain damage after stress in rats

Objective

The purpose of this study was to verify the presence of DNA brain lesion after acute stress in rats.

Method

Adult male Wistar rats were divided into 3 groups according to the stressor (control, forced swimming or restraint), and sampled at 2 time points: immediately or 1 week after stress. Trunk blood and the brain areas (prefrontal cortex, amygdala and hippocampus) were extracted for DNA analysis by the comet assay. The cells were classified according to the damage index and damage frequency based on the comet tail size.

Results

Immediately after the stress, DNA damage was detected in the amygdala area and in the hippocampus after restraint and forced swimming. In the prefrontal cortex, DNA was damaged after forced swimming. However, no alteration was seen in blood. Seven days after the stress, DNA damage was still identified in the hippocampus after forced swimming and restraint, whereas no alteration was detected in the other brain areas or in blood.

Conclusion

One week after a single stressful event, a reversible DNA damage was identified in the prefrontal cortex and in the amygdala, whereas DNA damage in the hippocampus still remained.     

Reduction in IV t-PA door to needle times using an Acute Stroke Triage Pathway

OBJECTIVE: To determine the effectiveness of an Acute Stroke Triage Pathway in reducing door to needle times in acute stroke treatment with IV t-PA. BACKGROUND: A previous study at our tertiary referral centre, examining IV t-PA door to needle times, was completed in 2000. The median door to needle time was beyond the recommended National Institute for Neurological Disorders and Stroke (NINDS) standard of 60 minutes. In November 2001, an Acute Stroke Triage Pathway was introduced in the emergency room (ER) to address this issue. The goal of this pathway was to rapidly identify patients eligible for treatment for IV t-PA, so that CT scans and lab studies could be arranged immediately upon ER arrival. Our hypothesis was that the Triage Pathway would shorten door to CT and door to needle times. DESIGN/METHODS: Using retrospective data, pre (n=87) and post (n=47) triage pathway times were compared. The door to CT time was reduced by 11 minutes (p=0.015) and door to needle time was reduced by 18 minutes (p=0.0036) in a subgroup of patients that presented directly to our hospital. CONCLUSIONS: These results indicate that the Acute Stroke Triage Pathway is effective in reducing Door to CT and Door to Needle Times in patients presenting directly to our ER. However, a majority of treatment times were still beyond NINDS recommendations. Stroke Centers require periodic review of their efficiency to ensure that target times are being obtained and may benefit from the use of an Acute Stroke Triage Pathway.     

Swimming induced vascular headache

We experienced 3 cases of characteristic headache induced by swimming. The first case was a 51-year-old woman who suffered from a sudden attack of throbbing headache in the parieto-temporal region, accompanied by nausea, after 20 minutes of swimming practice in a pool. The headache disappeared after about 3 hours of bed rest. However, soon after she began to practice swimming in the pool a few days later, throbbing headache in the same region recurred. Though the headache improved upon bed rest, dull pain was persistent till bed time that evening. Two days later, as she squatted down with alacrity at her home, she felt intense pain in the whole head as if struck with a hammer. She was seen at our neurological clinic the following day but was found to be neurologically normal with a blood pressure of 130/70 and a pulse rate of 78/min. Blood chemistry tests, hematology, head digital subtraction angiography, cerebral CT and X-ray films of the cervical spine were all normal. After swimming was prohibited, the prognosis was good and the patient experienced no episode of headache at all during the intervening period of 2 years and 5 months after the first visit. The second case was a 45-year-old man who felt throbbing headache affecting the whole head immediately after diving in a swimming pool. The headache improved after about 3 hours of bed rest. One week later, an attack of similar headache occurred immediately after a dive in the swimming pool.(ABSTRACT TRUNCATED AT 250 WORDS)     

大鼠CCA端端吻合在神经外科显微技术初学者训练中的价值

目的探讨大鼠颈总动脉(CCA)端端吻合在初学者显微技术训练中的价值。方法随机选取来自全国不同地区神经外科医生,且未进行过专门显微技术训练的初学者53名作为研究对象,同时随机选取健康雄性SD大鼠116只,分别行颈总动脉端端吻合术,并在吻合完成后检查即时血管通畅情况、吻合时间(自放置阻断夹至松开阻断夹)、吻合针数,并在24小时后检查次日血管通畅情况及存活率。结果 116只SD大鼠中,麻醉致死率约为6. 0%(7/116),大出血致死率约为4. 3%(5/116),最终有效研究对象104例,平均缝合6. 32针,平均用时64. 82 min,即时血管通畅率85. 6%(89/104),次日血管通畅率50. 0%(52/104),术后次日大鼠存活率100%(104/104)。结论大鼠颈总动脉端端吻合术后的即时、次日血管通畅率均较高,术后存活率高,可以作为未进行过专门显微技术训练的神经外科显微技术初学者训练的一种较好的动物模型。