脊髓损伤后Bcl-2抗神经元凋亡的研究

目的 通过研究Bcl-2(B-cell lymphoma/Leukemia-2)高表达转基因小鼠的脊髓损伤模型,探寻Bcl-2抗神经元凋亡,减轻脊髓继发性损害的作用.方法 将Bcl-2质粒转入小鼠的受精卵,培育Bcl-2转基因小鼠.转基因(A组)及同窝非转基因小鼠(B组)各9只随机(随机数字法)分为三组(1d,7 d,14 d),采用垂直打击脊髓(weight dropping,WD)方法,以2.5×3.0 g·cm致伤力致伤小鼠脊髓.另取3只非转基因小鼠设为假手术组(C组),仅打开椎板,暴露脊髓,不做打击.于术后1 d,7 d,14 d分别行:①小鼠后肢运动功能(BBB)评分;②HE染色观察;③原位末端标记法(TUNEL)染色.结果 打击后所有小鼠均出现双侧的后肢瘫痪,两周内BBB评分,A组(5.45±0.15)和B组(5.05±0.35),差异无统计学意义,P=0.67;HE染色提示A组小鼠的脊髓损伤程度较B组鼠要轻;原位末端标记法(TUNEL)染色表明A组小鼠TUNEL阳性的神经元(3.67±2.08/HP)明显小于B组小鼠(12±4/HP),P=0.033;积分光密度(Integration Optic Density,IOD)值(458.33±112.7)A组也明显小于B组小鼠(2436.33±228.01),P=0.000,其差异均具有统计学意义;C组未见明显的TUNEL阳性神经元.结论 Bcl-2蛋白是中枢神经系统内在的,重要的抗损伤因子,过表达Bcl-2能够抑制损伤后神经元的凋亡,从而减轻脊髓的损伤.     

大剂量甲基强的松龙对大鼠脊髓损伤后脊髓细胞凋亡的影响

目的：探讨大鼠脊髓损伤后应用大剂量甲基的强的松龙对脊髓损伤区细胞凋亡的影响。方法：将大鼠分为脊髓损伤应用大剂量甲基强的松龙治疗组（Ａ组）、单纯脊髓损伤组（Ｂ组）,损伤后１、３、７、１４、２８天驿脊髓损伤区进行细胞凋亡的检测（ＴＵＮＥＬ）以及Ｂｃｌ－２蛋白免疫反应表达的测定（免疫组化法）,采用计算机图像分析技术进行定量分析。结果：Ａ、Ｂ两组中均发现凋亡细胞及ＢＣＬ－２蛋白阳性表达,Ｂ组细胞凋亡率大于     

银杏叶提取物对大鼠脊髓损伤后神经凋亡细胞的影响

目的探讨银杏叶提取物对大鼠脊髓损伤后损伤区凋亡细胞的作用。方法60只大鼠分为正常组、损伤组和药物组,用Allen's改良法建立脊髓损伤的动物模型。银杏叶提取物通过腹腔注射,损伤后1、3、7、14、21 d取材,同时对大鼠损伤后双下肢功能的评估,对损伤区脊髓HE染色进行形态学观察,并用TUNEL法检测细胞凋亡,对结果进行统计分析。结果损伤区脊髓HE染色形态学观察表明,损伤组开始有片状出血,以后逐渐减少,到7 d全部消失。损伤组和药物组中均有凋亡细胞,损伤组的凋亡率大于药物组。结论银杏叶提取物能抑制或保护大鼠脊髓损伤后神经细胞的凋亡,从而减轻了脊髓损伤后的继发性损害。     

严重颅脑损伤后细胞凋亡状态及相关基因Bcl-2、Bax的表达

目的 探讨严重颅脑损伤后Bel-2、Bax基因表达与细胞凋亡的关系。方法 采用改进的Feeney自由落体损伤模型，应用分子生物学方法——脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(terminal-deoxynucleofidyl transferase mediated nick end labeling，TUNEL)以及免疫组织化学方法，对50只严重颅脑大鼠脑组织中的凋亡细胞和Bel-2、Bax免疫反应阳性的细胞进行观察、统计。结果 损伤组大鼠脑组织中凋亡细胞数明显高于未损伤组，Bel-2、Bax表达也有所增加，同时，损伤组Bax／Bel-2阳性细胞数的比值明显大于未损伤组。结论 Bel-2和Bax表达的比例对严重颅脑损伤后细胞凋亡可能起重要的调控作用。     

脊髓损伤大鼠核因子kappa B与细胞凋亡及肢体功能间的相关性分析

Objective To investigate the relationship of nuclear factor kappa B(NF-κB),Bcl-2 and Bax with limb function after acute spinal cord injury in rats. Methods Forty-eight rats were divided at random into a control group and an experimental group with 24 rats in each.The spinal cords of the rats in the experimental group were injured at the T8,9,10 level through moderate compression.Four hours,8 h,and 1,3,7 and 14 days after the injury,4 rats were selected randomly from each group and graded with a BBB score.They were then sacrificed and their spinal cords were collected.Immunohistochemical measurements were used to observe the expression of NF-κB, Bcl-2 and Bax. Results NF-κB,Bcl-2 and Bax were observed in the injured spinal nerve cells of rats in the exper imental group but were absent in the control group.After injury,the expression of these factors increased at first and then decreased.BBB scores for limb function increased gradually.No correlation was found between the changes in NF-κB and Bcl-2,but the expression of NF-κB was positively correlated with that of Bax.There was negative correla tion between NF-kB levels and BBB scores,and between NF-kB levels and the ratio of Bcl-2 to Bax. Conclusion In rats,there is a close negative correlation between NF-kappa B levels,the ratio of Bcl-2/Bax and limb function after acute spinaI cord iujury.     

大鼠脊髓损伤后细胞凋亡特点及其与一氧化氮合酶表达的相关性

目的探讨脊髓损伤后细胞凋亡特点与诱生型一氧化氮合酶(iNOS)表达规律及二者之间的关系.方法成年大鼠32只,随机分为8组(每组4只)单纯椎板切除对照组和脊髓损伤后7个时间点处死组(4、8 h和1、3、7、14、21 d).用免疫组化染色检测iNOS、p53阳性细胞,原位末端标记法(TUNEL)标记凋亡细胞.另取大鼠32只进行同样分组,用Western blot analsis法检测iNOS表达.结果免疫组化结果显示单纯椎板切除对照组神经元、神经胶质细胞、室管膜细胞、血管内皮细胞极少量表达iNOS;损伤后8 h上述细胞iNOS表达增加,7 d达高峰,14 d仍较明显,21 d降低.p53表达及TUNEL标记阳性细胞的时间分布特点与iNOS相似,7d达高峰,阳性细胞以白质中胶质细胞为主,主要分布于损伤部及相邻区.iNOS的Western blot灰度扫描数值时间变化曲线与免疫组化阳性细胞率时间变化曲线一致.iNOS表达与神经细胞凋亡指数及p53表达存在正相关(r＝0.854,P＜0.01;r＝0.951,P＜0.01).结论大鼠脊髓损伤后iNOS与p53表达增强,凋亡神经细胞大量出现.iNOS表达与脊髓损伤后神经细胞凋亡指数间存在正相关.     

急性脊髓损伤后并发重度低钠血症的临床分析

目的:探讨急性脊髓损伤后并发重度低钠血症的临床特点和治疗方法。方法:1997年10月至2005年12月间我科共收治358例急性脊髓损伤患者,其中121例(33.8%)发生低钠血症(血清钠<135mmol/L),对其中8例重度低钠血症(血清钠<120mmol/L)患者资料进行回顾性分析。结果:8例中男5例,女3例,年龄25～51岁,平均36岁,均因车祸或高处坠落致伤,7例为颈脊髓损伤,1例为胸髓损伤,均为完全性脊髓损伤(AsiaA级),2例合并有轻度闭合型颅脑损伤。平均出现低钠血症时间为伤后(5.7±1.8)d,最低血清钠浓度为(111.6±4.0)mmol/L,出现低钠高峰时间为伤后(8.1±2.0)d,低钠持续时间(20.9±5.9)d。8例均给予深静脉插管,行中心静脉压监测。6例经限水、补钠等治疗后低钠血症纠正,2例低钠血症进行性加重,出现呼吸、循环衰竭死亡。2例合并有肺部感染。结论:急性脊髓损伤后早期易并发低钠血症,由于其症状隐匿,常不被重视,一旦出现重度低钠血症,治疗非常棘手,死亡率高。     

脊髓损伤对睾酮水平和Leydig细胞凋亡影响的实验研究

目的探讨脊髓损伤诱发低睾酮血症的机制。方法成年健康SD大鼠20只,随机分为假手术组和脊髓损伤组,手术后14 d取血清以放免法检测睾酮(T)和黄体生成素(LH)水平,以脱氧核糖核酸末端转移酶介导的dUTP的缺口末端标记技术(TUNEL)原位检测Leydig细胞凋亡,以Percoll连续密度梯度法提取Leydig细胞后采用流式细胞仪(FACS)定量检测Leydig细胞凋亡情况。结果术后14 d,脊髓损伤组大鼠血清T水平较假手术组显著降低(P<0.05),LH水平较假手术组显著升高(P<0.05),TUNEL显示睾丸间质中Leydig细胞发生凋亡,脊髓损伤组凋亡Leydig细胞比例(46%)显著高于假手术组(17.67%)(P<0.05)。结论诱发Leydig细胞凋亡可能是脊髓损伤导致低睾酮血症发生的重要机制之一。     

脊髓损伤后神经源性膀胱的个体化康复护理

目的 探讨脊髓损伤后神经源性膀胱的最佳护理方法。方法 60例脊髓损伤后排尿障碍患者随机分为2组，分别采用传统护理法和个体化护理法，比较两组患者自主排尿时问和泌尿系统并发症发生率的差别，从而找到适宜的护理方法。结果 个体化护理组患者自主排尿早，并发症少。结论 针对脊髓损伤后神经源性膀胱的不同特点，实施个体化护理措施，更有利于膀胱功能恢复。     

Exogenous administration of glial cell line-derived neurotrophic factor improves recovery after spinal cord injury

The aim of present study was to examine whether systemically delivered glial cell-derived neurotrophic factor (GDNF) was beneficial in reversing the spinal cord injury (SCI) in a spinal cord compression model. Rats were divided into three major groups: (1) sham operation (laminectomy only); (2) laminectomy+SCI+normal saline (1ml/kg, i.v.); (3) laminectomy+SCI+GDNF (50ng/kg, i.v.). Spinal cord injury was induced by compressing the spinal cord for 1min with an aneurysm clip calibrated to a closing pressure of 55g. GDNF or saline was administered immediately after SCI via the tail vein. Behavioral tests of motor function measured by maximal angle an animal could hold to the inclined plane were conducted at days 1-7 after SCI. The triphenyltetrazolium chloride staining and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling assay were also conducted after SCI to evaluate spinal cord infarction and apoptosis, respectively. Both GDNF and vascular endothelial growth factor (VEGF) in the injured spinal cord were assayed by immunofluorescence. It was found that systemically delivered GDNF, but not vehicle solution, significantly attenuated the SCI-induced hind limb dysfunction and spinal cord infarction and apoptosis. Both GDNF and VEGF could be detected in the injury spinal cord after GDNF, but not vehicle solution, therapy. The results indicate that GDNF treatment may be beneficial in reversing hind limb dysfunction by reducing spinal cord infarction and apoptosis in a spinal cord compression model.     

Adjusting expectations after spinal cord injury across global settings: A commentary

Purpose. The objective of this paper is to stimulate thought and discussion as to how best to set rehabilitation goals to maximize activities and participation of persons with spinal cord injury, across global settings where circumstances and environments may be widely different.

Method. A review of literature and commentary are presented. Three points are articulated: (1) rehabilitation professionals need to understand factors that impact upon activities and participation, and need measurement tools that report these factors, in order to better appreciate outcomes in different settings, (2) rehabilitation professionals generally set goals with patients, but current measures of activities and participation do not indicate when or why maximal achievable function is sometimes not chosen, and (3) we need to develop realistic expectations for activities and participation after SCI in settings where current standard outcome chart targets are not feasible, due to socio-economic circumstances.

Conclusions. A standardized approach to reporting measures of activities and participation, along with factors that influence these scores, is required for purposes of comparing rehabilitation outcomes in settings of differing socio-economic environments. In regards to spinal cord injury rehabilitation, an accepted standard of setting achievable rehabilitation goals is required for each level of complete spinal cord injury that could apply in various global settings.     

Adjusting expectations after spinal cord injury across global settings: a commentary

Purpose. The objective of this paper is to stimulate thought and discussion as to how best to set rehabilitation goals to maximize activities and participation of persons with spinal cord injury, across global settings where circumstances and environments may be widely different.

Method. A review of literature and commentary are presented. Three points are articulated: (1) rehabilitation professionals need to understand factors that impact upon activities and participation, and need measurement tools that report these factors, in order to better appreciate outcomes in different settings, (2) rehabilitation professionals generally set goals with patients, but current measures of activities and participation do not indicate when or why maximal achievable function is sometimes not chosen, and (3) we need to develop realistic expectations for activities and participation after SCI in settings where current standard outcome chart targets are not feasible, due to socio-economic circumstances.

Conclusions. A standardized approach to reporting measures of activities and participation, along with factors that influence these scores, is required for purposes of comparing rehabilitation outcomes in settings of differing socio-economic environments. In regards to spinal cord injury rehabilitation, an accepted standard of setting achievable rehabilitation goals is required for each level of complete spinal cord injury that could apply in various global settings.     

早期功能锻炼对脊髓损伤患者骨质疏松的预防作用

目的探讨早期肢体被动功能锻炼对脊髓损伤患者骨质疏松的预防作用。方法将26例外伤性脊髓损伤所致截瘫患者随机分为实验组和对照组,每组13例,实验组进行早期肢体功能锻炼,对照组按照外伤瘫痪患者护理常规进行。常规x线片观察两组患者锻炼前后（入组时和4个月后）腰椎及股骨近端的松质骨密度变化,同时分别检测两组患者锻炼前后（入组时和4个月后）血清碱性磷酸酶、血钙、尿Ca／Cr及尿羟脯氨酸含量。结果腰椎及股骨近端的松质骨密度实测值与入组时比较,锻炼后（4个月后）两组均有下降趋势,但均无统计学意义（P〉0．05）,两组间差异也无统计学意义（P〉0．05）。与入组时比较,锻炼后（4个月）实验组血清碱性磷酸酶无统计学差异（P〉0．05）,血钙、尿Ca／Cr及尿羟脯氨酸均有统计学意义（P〈0．05）;对照组血清碱性磷酸酶血钙、尿Ca／Cr及尿羟脯氨酸均无统计学意义（P〉0．05）。锻炼后（4个月）两组间除血清碱性磷酸酶差异无统计学意义（P〉0．05）,血钙、尿Ca／Cr及尿羟脯氨酸差异均有统计学意义（P〈0．05）。结论早期肢体功能锻炼对脊髓损伤患者骨质疏松具有一定的预防作用。     

脊髓损伤大鼠核因子kappa B与细胞凋亡及肢体功能间的相关性分析

目的研究急性脊髓损伤（ASCI）大鼠核因子kappa B（NF-κB）与细胞凋亡及肢体功能间的相关性。 方法共选取48只雌性SD大鼠,将其随机分为实验组及对照组。2组大鼠均采用手术切除椎板,对照组术中未进行脊髓压迫处理,实验组大鼠术中采用压迫法制作脊髓中度损伤模型。2组均于术后第4 h、8 h、1 d、3 d、7 d及14 d时各随机选取4只大鼠,参照BBB评分标准对其后肢功能进行评定,待评分结束后处死大鼠并提取脊髓标本,采用免疫组织化学染色法检测各组大鼠脊髓标本内NF-κB、Bcl-2及Bax表达水平。 结果实验组大鼠受损脊髓神经细胞中有NF-κB、Bcl-2及Bax表达,且表达水平随术后时间延长呈现先增高、后降低过程;大鼠肢体功能BBB评分随时间延长而逐渐升高。经相关性分析发现,随着术后时间延长,实验组大鼠NF-κB表达量与Bcl-2表达量无明显相关性（P＞0.05）,与Bax表达量有正相关性（P＜0.05）,与Bcl-2/Bax比值及BBB评分有负相关性（P＜0.05）。 结论ASCI大鼠NF-κB表达与Bcl-2/Bax比值及肢体运动功能均具有显著负相关性。     

大鼠脊髓不同平面损伤后神经源性膀胱模型的尿流动力学检查

目的观察大鼠不同平面脊髓损伤（SCI）后尿流动力学的改变，探讨不同平面SCI引起神经源性膀胱的病理生理机制。方法选择健康雌性SD大鼠80只，分为骶上脊髓（胸腰段）损伤组（30只）、骶髓损伤组（30只）及对照组（20只）。采用脊髓横断法制备大鼠脊髓不同平面损伤后神经源性膀胱模型。分别于动物模型制作前及制作后第20天行尿流动力学检查，评价大鼠的膀胱功能。结果骶上脊髓损伤组逼尿肌漏尿点压较对照组、骶髓损伤组明显升高，差异有统计学意义（P〈0．01），骶髓损伤组逼尿肌漏尿点压较对照组显著降低，差异有统计学意义（P〈0．01）；骶上脊髓损伤组膀胱最大容量、膀胱顺应性较对照组、骶髓损伤组明显降低，差异有统计学意义（P〈0．01），骶髓损伤组膀胱最大容量、膀胱顺应性较对照组显著增加，差异有统计学意义（P〈0．01）。结论尿流动力学检查为神经源性膀胱的早期诊断及选择个体化治疗方案提供了一种有效的检查手段。     

外伤致脊髓损伤患者的康复期护理

目的:探讨外伤后脊髓损伤患者康复期的护理要点。方法:通过康复治疗小组进行治疗及心理支持,对108例外伤后脊髓损伤康复期患者实施各种康复护理措施。对可能出现的各种并发症,护士及早的康复介入,循序渐进,进行防治。结果:经过康复护理的早期介入,提高了脊髓损伤患者的生活质量,调动了脊髓损伤患者的主动性,发挥其残存功能,帮助他们重归家庭和社会。结论:外伤后脊髓损伤患者的康复,要尽早开始,减少并发症,并依靠患者自身的努力,改善生活自理能力。     

Peripheral noxious stimulation reduces withdrawal threshold to mechanical stimuli after spinal cord injury: Role of tumor necrosis factor alpha and apoptosis

We previously showed that peripheral noxious input after spinal cord injury (SCI) inhibits beneficial spinal plasticity and impairs recovery of locomotor and bladder functions. These observations suggest that noxious input may similarly affect the development and maintenance of chronic neuropathic pain, an important consequence of SCI. In adult rats with a moderate contusion SCI, we investigated the effect of noxious tail stimulation, administered 1 day after SCI on mechanical withdrawal responses to von Frey stimuli from 1 to 28 days after treatment. In addition, because the proinflammatory cytokine tumor necrosis factor alpha (TNFα) is implicated in numerous injury-induced processes including pain hypersensitivity, we assessed the temporal and spatial expression of TNFα, TNF receptors, and several downstream signaling targets after stimulation. Our results showed that unlike sham surgery or SCI only, nociceptive stimulation after SCI induced mechanical sensitivity by 24 h. These behavioral changes were accompanied by increased expression of TNFα. Cellular assessments of downstream targets of TNFα revealed that nociceptive stimulation increased the expression of caspase 8 and the active subunit (12 kDa) of caspase 3, indicative of active apoptosis at a time point consistent with the onset of mechanical allodynia. In addition, immunohistochemical analysis revealed distinct morphological signs of apoptosis in neurons and microglia at 24 h after stimulation. Interestingly, expression of the inflammatory mediator NFκB was unaltered by nociceptive stimulation. These results suggest that noxious input caudal to the level of SCI can increase the onset and expression of behavioral responses indicative of pain, potentially involving TNFα signaling.     

减重步行激活SCI大鼠腰膨大前角神经元效应的激光共聚焦镜检研究

目的 探讨减重平板步行训练(BWSTT)促进脊髓损伤(SCI)功能恢复的脊髓可塑性机制.方法 将84只成年雌性Wistar大鼠随意分为假手术组(Sham组,n=18)、胸髓横断模型组(SCI组,n=33)和减重平板步行训练组(BWSTT组,n=33),每组大鼠再随机分为7,15,45 d 3个亚组.建立大鼠胸髓完全横断模型,观察BWSTT对SCI大鼠后肢运动功能的影响.采用激光共聚焦镜检免疫荧光双标技术.研究SCI大鼠腰髓前角腹内侧区神经元酪氨酸蛋白激酶受体(EphA4)、囊泡膜谷氨酸转运体2(VGluT2)的表达,以及两者双标免疫阳性细胞比率的变化.结果 BWSTT组大鼠的后肢运动功能较SCI组大鼠明显改善.胸髓横断大鼠BWSTT后,腰髓前角腹内侧区EphA4和VGIuT2的表达以及EphA4/VGhT2双标免疫阳性细胞比率均显著增加.结论 BWSTT通过增强胸髓横断大鼠腰髓前角神经元EphA4/VGIuT2的表达,促进后肢运动功能的恢复.     

减重步行激活SCI大鼠腰膨大前角神经元效应的激光共聚焦镜检研究

目的 探讨减重平板步行训练(BWSTT)促进脊髓损伤(SCI)功能恢复的脊髓可塑性机制.方法 将84只成年雌性Wistar大鼠随意分为假手术组(Sham组,n=18)、胸髓横断模型组(SCI组,n=33)和减重平板步行训练组(BWSTT组,n=33),每组大鼠再随机分为7,15,45 d 3个亚组.建立大鼠胸髓完全横断模型,观察BWSTT对SCI大鼠后肢运动功能的影响.采用激光共聚焦镜检免疫荧光双标技术.研究SCI大鼠腰髓前角腹内侧区神经元酪氨酸蛋白激酶受体(EphA4)、囊泡膜谷氨酸转运体2(VGluT2)的表达,以及两者双标免疫阳性细胞比率的变化.结果 BWSTT组大鼠的后肢运动功能较SCI组大鼠明显改善.胸髓横断大鼠BWSTT后,腰髓前角腹内侧区EphA4和VGIuT2的表达以及EphA4/VGhT2双标免疫阳性细胞比率均显著增加.结论 BWSTT通过增强胸髓横断大鼠腰髓前角神经元EphA4/VGIuT2的表达,促进后肢运动功能的恢复.     

Managing fatigue following spinal cord injury: A qualitative exploration

Purpose. To identify, from the perspectives of people with spinal cord injury (SCI), (a) appropriate components of a fatigue management programme; and (b) important outcomes or indicators of success.

Method. Collaborative, qualitative methodology comprising four focus groups undertaken simultaneously in Kelowna, Prince George, Vancouver and Victoria, British Columbia, Canada. Participants included a purposive sample of 21 men and women with complete and incomplete SCI of high and low tetraplegia and paraplegia. Two family members, two care-providing assistants and four occupational therapists provided additional information (total n = 29). Interpretive data analysis identified common themes addressing each research question.

Results. Building on those strategies they perceived to facilitate coping with fatigue, the participants identified 10 components of a helpful fatigue management programme. Dimensions of ‘successful’ outcomes from such a programme reflected quality of life concerns: enabling people with SCI to do the things they value, enhancing their sense of control over their lives, reducing pain and helplessness, increasing motivation and enhancing relationships strained by fatigue.

Conclusions. This study identifies many of the necessary elements of a fatigue management programme to meet the specific needs of people with SCI; and ascertains important indicators of a successful programme from the perspectives of those who must live with the outcomes.