Morphological and electrophysiological evidence for regeneration of transected spinal cord fibers and restoration of motor functions in adult rats

The evoked potentials are regarded as an efficientindex to evaluate the functional status of a nervoussystem[1]. When stimulating the motor area of cerebralcortex with transcranial magnetic stimulation, elec-tronic signals can be obtained at the spinal co…     

基因重组碱性成纤维细胞生长因子对大鼠脊髓损伤神经保护作用

目的 :探索碱性成纤维细胞生长因子 (bFGF)对大鼠脊髓损伤的神经保护作用 方法 :将吸入bFGF的胶原蛋白海绵或空白海绵贴敷于大鼠脊髓损伤处 ,术后 1、2、3周 ,对大鼠机能进行评分 ,并对大脑运动皮质进行电镜观察分析 结果 :术后 1、2、3周 ,bFGF组大鼠运动评分均明显优于对照组 (Ρ <0 .0 5) ,运动皮质电镜结果显示bFGF组线粒体、内质网轻度肿胀 ,神经毡结构正常 ,无明显胶质细胞增生 程度较对照组明显减轻 结论 :bFGF对大鼠脊髓受损神经纤维起源脑区—运动皮质的神经细胞具有明显的保护作用 ,进而使大鼠运动功能受损明显减轻     

Neural stem cell transplantation in the repair of spinal cord injury

Neural stem cells are a pronising candidate for neural transplantation aimed at neural cell replacement and repair of the damaged host central nervous system (CNS). Recent studies using neural stem cells have shown that implanted neural stem cells can effectively incorporate into the damaged CNS and differentiate into neurons, astrocytes, and oligodendrocytes. The recent explosion in the field of neural stem cell research has provided insight into the inductive factors influencing neural stem cell differentiation and may yield potential therapies for several neurological disorders, including spinal cord injury. In this review, we summarize recent studies involving neural stem cell biology in both rodents and humans. We also discuss unique advantages and possible mechanisms of using neural stem cell trans plantation in the repair of spinal cord injury.     

Protective paracrine effect of mesenchymal stem cells on cardiomyocytes

Objective: The aim of this study was to test the protective effect of mesenchymal stem cells (MSCs) on cardiomyocytes in vitro and to investigate the anti-apoptotic signaling pathway. Methods: MSCs from Sprague-Dawley (SD) rats were separated and cultured. MSC medium was collected from MSCs cultured in serum-free Dulbecco’s modified eagle medium (DMEM) under hypoxia. Cultured cardiomyocytes from neonatal SD rats were exposed to hypoxia/reoxygenation (H/R) and treated with MSC medium. The apoptotic cardiomyocytes were stained with Annexin-V-fluorescein isothiocyanate (FITC), Hoechst 33342 and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL). The mitochondrial transmembrane potential of cardiomyocytes was assessed using a fluorescence microscope. The expression of Bcl-2, Bax, cytochrome C, apoptosis-induced factor (AIF), and caspase-3 was tested by Western blot analysis. Results: Our data demonstrated that MSC medium reduced H/R-induced cardiomyocyte apoptosis, increased the Bcl-2/Bax ratio, and reduced the release of cytochrome C and AIF from mitochondria into the cytosol. Conclusion: MSCs protected the cardiomyocytes from H/R-induced apoptosis through a mitochondrial pathway in a paracrine manner.     

积雪草苷调控miR-342-5p/AQP3轴保护IL-1β诱导的软骨细胞损伤

目的 探讨积雪草昔(Ass)对白细胞介素1B(IL-1B)诱导的软骨细胞损伤的影响及作用机制。方法 不同 浓度的Ass作用IL-1B诱导软骨细胞24 h,流式细胞仪检测细胞凋亡,酶联免疫吸附法检测细胞培养上清液中白细 胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)水平,实时荧光定量PCR检测细胞中miR-342-5p和水通道蛋白3 (AQP3)mRNA表达水平,Western Blot法检测B淋巴细胞瘤-2 (Bcl-2)、B淋巴细胞瘤-2相关蛋白(Bax)和AQP3蛋 白表达水平。双荧光素酶报告基因实验验证miR-342-5p与AQP3靶向关系。转染miR-342-5p抑制剂或AQP3过表达载体至软骨细胞,上述相同方法检测抑制miR-342-5p表达或AQP3过表达对IL-1β诱导的软骨细胞凋亡及IL- 6和TNF-α表达的影响。结果 Ass可抑制IL-1β诱导的软骨细胞凋亡率、Bax蛋白、IL-6和TNF-α表达及miR- 342-5p表达(P <0. 05),促进Bcl-2蛋白、AQP3 mRNA和蛋白表达(P <0.05)。miR-342-5p在软骨细胞中负调控 AQP3表达。抑制miR-342-5p或AQP3过表达后,IL-1&诱导的软骨细胞凋亡率、Bax蛋白、IL-6和TNF-α表达降低 (P <0. 05) ,Bcl-2蛋白表达升高(P <0.05)。抑制AQP3表达逆转了抑制miR-342-5p对IL-1β诱导的软骨细胞凋 亡、Bax和Bcl-2蛋白及IL-6和TNF-α表达的影响。结论 Ass可抑制IL-1β诱导软骨细胞凋亡和炎症反应,其可 能通过调控miR-342-5p/AQP3保护软骨细胞损伤。     

白藜芦醇上调GDF-15调控PI3K/Akt/Bcl-2信号通路对急性心肌梗死大鼠的保护作用

为考察白藜芦醇对心肌缺血的保护机制,采用冠状动脉前降支结扎再灌注制备大鼠心肌缺血再灌注(I/R)模型,运用试剂盒考察血清酶学指标、NBT染色考察心肌梗死率、蛋白质印迹法(Western blotting) 考察GDF-15、PI3K、Akt、p-Akt、Bcl-2、Bax、caspase-9和caspase-3的蛋白相对表达量。结果表明:白藜芦醇对心肌缺血再灌注大鼠的心肌保护作用与通过对GDF-15的上调来激活PI3K/Akt信号通路,抑制心肌细胞线粒体凋亡作用机制有关。     

HCAP1基因对Raji细胞凋亡及凋亡相关蛋白Bax/Bcl - 2的作用

目的：研究外源HCAP1基因产物对Burkitt淋巴瘤细胞系Raji细胞凋亡及凋亡相关蛋白Bax／Bcl-2的作用。方法：用脂质体介导的基因转移方法，把外源HCAP1基因转染到Raji细胞中，用流式细胞术和Hochest 33258荧光染色检测细胞凋亡；用Western blot检测外源HCAP1基因对凋亡相关蛋白Bax和Bcl-2蛋白表达的调节。结果：外源HCAPl基因导入Raji细胞24、48和96h后，细胞凋亡率增加。Western blot显示Bax／Bel-2蛋白表达比值明显增高。结论：转染外源性HCAP1基因可诱导Raji细胞凋亡，使Bax／Bcl-2蛋白表达比例上调可能是HCAP1诱导凋亡的机制之一。     

灯盏花注射液抗新生鼠缺氧缺血脑损伤的药效作用

为观察灯盏花注射液治疗新生鼠缺氧缺血脑损伤的药物疗效及其药理作用机制,采用新生7日龄大鼠缺氧缺血性脑损伤模型,设立假手术组、缺氧缺血脑损伤模型组、灯盏花注射液治疗10,20,40 mg/kg剂量组、无菌注射用水对照组.采用硫堇染色、免疫组织化学染色的方法测定各实验组海马CA1区神经元密度、组织学分级、凋亡相关基因bcl-2和bax的表达情况,并计数各时间点Bcl-2和Bax蛋白免疫阳性细胞数目及积分光密度值.假手术组,大鼠海马CA1区无锥体细胞缺失,未见明显阳性细胞.与假手术组比较,缺氧缺血脑损伤模型组、无菌注射用水对照组Bcl-2,Bax蛋白表达均于3 d时达到高峰(与其余各时间点比较差别有显著意义p0.05),神经元密度明显降低,组织学分级显著增高,积分光密度值增加.灯盏花治疗组,与无菌注射用水对照组比较,Bcl-2阳性表达进一步增加,积分光密度值增加;而Bax阳性表达则减少,积分光密度值降低;神经元密度显著高于对照组,组织学分级明显降低.灯盏花注射液可能是通过上调Bcl-2表达,抑制Bax表达,减轻缺氧缺血引起的神经元凋亡及迟发性神经元死亡.     

铅对小鼠睾丸生精细胞凋亡及Caspase-3,Bcl-2和Bax基因表达的影响

为了研究铅致小鼠睾丸生精细胞凋亡及其对Caspase-3,Bax和Bcl -2基因表达的影响.25只雄性小鼠随机分成5组:0h对照组、12h染铅组、24h染铅组、36h染铅组和48h染铅组,每组5只,实验组按照100mg/kg的剂量腹腔注射醋酸铅溶液,5组小鼠分别于0h、12h、24h、36h、48h后脊柱拉断处死,取其睾丸组织,用苏木精-伊红染色及免疫组化技术检测睾丸组织细胞凋亡及凋亡相关蛋白Caspase -3,Bcl -2和Bax的表达.结果显示:细胞凋亡相关基因Caspase -3,Bcl -2和Bax的表达较对照组差异显著,Caspase -3和Bax在染铅毒后表达量一直上升,Bcl -2的表达量一直下降,差异性随染铅时间的不同而有变化.铅负荷至一定程度时可致小鼠睾丸细胞凋亡,且呈一定量效关系(P＜0.01),这种作用可能与Bcl -2基因低表达和Caspase-3与Bax基因高表达有关.     

Chondroitinase ABC promotes functional recovery after spinal cord injury

The inability of axons to regenerate after a spinal cord injury in the adult mammalian central nervous system (CNS) can lead to permanent paralysis. At sites of CNS injury, a glial scar develops, containing extracellular matrix molecules including chondroitin sulphate proteoglycans (CSPGs). CSPGs are inhibitory to axon growth in vitro, and regenerating axons stop at CSPG-rich regions in vivo. Removing CSPG glycosaminoglycan (GAG) chains attenuates CSPG inhibitory activity. To test the functional effects of degrading chondroitin sulphate (CS)-GAG after spinal cord injury, we delivered chondroitinase ABC (ChABC) to the lesioned dorsal columns of adult rats. We show that intrathecal treatment with ChABC degraded CS-GAG at the injury site, upregulated a regeneration-associated protein in injured neurons, and promoted regeneration of both ascending sensory projections and descending corticospinal tract axons. ChABC treatment also restored post-synaptic activity below the lesion after electrical stimulation of corticospinal neurons, and promoted functional recovery of locomotor and proprioceptive behaviours. Our results demonstrate that CSPGs are important inhibitory molecules in vivo and suggest that their manipulation will be useful for treatment of human spinal injuries.     

Dick氏内固定器治疗胸腰椎骨折脱位并脊髓损伤

为寻求和解决胸腰椎骨折脱位并脊髓损伤的有效疗法,用DICk氏内固定器治疗53例患者,并随访．对临床资料进行总结分析。结果表明,Dick氏内固定系统符合脊柱的生物力学原理,可使固定节段具有高度的内在稳定性,且具有操作简便、安全、短节段固定,合并症少等优点。     

Effect of Jiaji electroacupuncture in transected rat spinal cord

We evaluated the effect of Jiaji electroacupuncture on cell proliferation and the expression of markers of endogenous neural stem cell activation after complete spinal cord transection. Female Wistar rats were assigned to 4 groups (n = 24 each): a sham-operated group, a control group, a Jiaji electroacupuncture group, and a Jiaji electroacupuncture preconditioning group. Motor function was significantly improved in the acupuncture groups compared to the control group at 7 and 14 d. Numbers of bromodeoxyuridine (BrdU)-, nestin-, and glial fibrillary acidic protein (GFAP)-positive cells were significantly greater in the acupuncture groups than in the controls at each time point. Expression of nestin and GFAP mRNA was significantly higher in the acupuncture groups than in the controls at each time point. Thus, Jiaji electroacupuncture and preconditioning may promote the proliferation of endogenous neural stem cells after spinal cord transection.     

Preliminary animal studies on observation of injured spinal cord with intraoperative ultrasound backscatter microscopy

Many studies have shown that strategies of nerve regeneration and cell-based transplantation are valid based on animal models of spinal cord injury (SCI).To apply these strategies and bridge spinal cord defects,the identification and precise localization of lesions during spinal cord surgery is necessary.The aim of the present experiment was to evaluate the capabilities of ultrasound backscatter microscopy (UBM) in identifying morphologic changes after SCI.After laminectomy,high-resolution ultrasound images of the spinal cord were obtained in one normal and seven spinal cord-injured adult Wistar rats using a UBM system with a 55-MHz center frequency scanner.Comparison between histoanatomic and UBM images was also performed.The results showed that UBM can identify cysts after the experimental SCI is removed in adult rats.In addition,the glial scar formed in secondary injury showed obvious hyperechoic speckle in the UBM image and correlated with the histoanatomic image.UBM has obvious clinical value in nerve regeneration and cell-based transplantation strategies in injured spinal cords.     

体育功能锻炼对截瘫患者康复影响及评价分析

目的:探讨体育功能锻炼对截瘫患者后期康复治疗的影响及康复评价.方法:76例在脊髓损伤后出现截瘫患者,进行体育功能锻炼的康复治疗,追踪记录,采用美国脊髓损伤学会(ASIA)脊髓损伤神经功能分类国际标准评价康复效果.结果:经统计,胸腰髓不完全损害者康复锻炼前后ASIA损伤评分运动、感觉评分均有显著意义(P<0.05).胸腰髓完全损伤者运动、感觉ASIA评分好转较明显(P<0.05).结论:体育功能锻炼能帮助截瘫患者后期恢复,显著改善和提高患者的综合功能.     

表儿茶素没食子酸酯对人鼻咽癌C666-1细胞凋亡的影响

该文观察ECG在人鼻咽癌细胞株C666-1增殖与凋亡中的作用,并初步探讨其机制.人鼻咽癌C666-1细胞给予不同浓度ECG(0~100μmol/L)处理48 h后,采用CCK-8法检测细胞增殖水平,使用TUNEL法和流式细胞仪检测细胞凋亡率,利用Western blotting法检测Bax、Bcl-2、Caspase 3等凋亡相关蛋白的表达变化.结果表明,25μmol/L以上浓度的ECG可抑制C666-1细胞增殖,诱导细胞凋亡;并且ECG可浓度依赖性增加Bax/Bcl-2蛋白比值和剪切的Caspase 3蛋白水平.以上结果表明ECG可抑制人鼻咽癌细胞株C666-1增殖,并诱导细胞凋亡.ECG的作用与其增加Bax/Bcl-2相对蛋白比值进而激活caspase-3有关.     

乌鳢尾部神经分泌系统组织化学和HRP法研究

采用辣根过氧化物酶(HRP)法结合乙酰胆碱酯酶(AChE)和单胺氧化酶(MAO)组化方法,对乌鳢尾部神经分沁系统进行了研究。结果如下: 1.在尾部脊髓中可以观察到一些HRP标记的神经分泌细胞和HRP标记末梢。有些标记末梢终止于脊髓中央管,少量终止于腹面脊膜。尾垂体中也有许多HRP标记末梢。2.神经分泌细胞细胞体中的HRP颗粒在电镜下呈圆形、短管状和哑铃状。3.神经分泌细胞体显示AChE活性,并定位于核膜、粗面内质网上。4.神经分泌细胞体周围尚显示MAO活性。     

苦丁茶对MCF-7人乳腺癌细胞的体外抗癌效果

对市售的苦丁茶进行MCF-7人乳腺癌细胞体外抗癌效果和体内抗转移效果评价.通过MTT实验、DAPI荧光染色分析和RT-PCR分析说明其体外抗癌效果.200μg/mL苦丁茶(81%抑制率)水提物表现出对MCF-7人乳腺癌细胞较强的生长抑制效果.200μg/mL比100和50μg/mL苦丁茶水提物具有更强的细胞诱导凋亡效果,Bax,caspase-3和caspase-9的mRNA表达得到增强,Bcl-2表达减弱.苦丁茶水提物处理后炎症相关因子NF-κB,iNOS和COX-2表达减弱,展示了苦丁茶的抗炎特性.由此得出,一定质量浓度的苦丁茶具有良好的抗癌预防效果.     

Same impact momentum causes different degrees of spinal cord injury

The weight-drop impact is widely used in making animal model of spinal cord injury （SCI）. But there has not yet been an appropriate unit for the quantification of the impact. In this study, we compared the degrees of the spinal cord injury caused by weight-drop impact with the same momentum but different combinations of drop weight and drop height, in order to test whether ＂momentum＂ is capable of being the unit for the quantification of weight- drop impact. Thirty adult rats were randomly allocated to three groups and were injured with 5 g-10 cm （group A）, 10g-2.5 cm （group B） and 15 g-l.ll cm （group C） impacts with the same momentum to the spinal cord, respectively. Open-field locomotor function was evaluated using the Basso-Beattie-Bresnahan （BBB） locomotor rat- ing scale. The percentage of spared tissue area （STA） at the epicenter, and 500, 1000 and 1500 gm from the epicenter was calculated using serial sections stained by hematoxylin and eosin. As a result, the behavioral recovery （BBB scores） and the STA percentage were similar in group B and group C. However, the BBB score in group A was significantly lower than that in groups B and C at the same time point post injury. The STA percentage was signifi- cantly less and the lesion/cavity length was significantlygreater in group A than in groups B and C. These suggested that the 5 g-10 cm weight-drop impact, compared with the other two impacts with different weights and heights, caused a greater damage of the spinal cord when the momentum was the same. So, these impacts with the same momentum but different weights and drop heights cause different degrees of spinal cord injury. Momentum alone is inadequate to be the unit for the qualification of weight- drop impact and to be used to predict the extent of injury.     

内耳免疫反应诱导Bcl-2和Bax表达研究

探讨内耳免疫反应过程是否引起细胞凋亡以及Bcl-2和Bax的表达情况。选用雌性白色豚鼠16只，随机分为实验组和对照组各8只，以钥孔虫戚血蓝蛋白（KLH）全身免疫后，实验组以相同抗原进行内耳免疫，对照组内耳注射等量的磷酸盐缓冲生理盐水（PBS），在内耳免疫5d后处死动物，取内耳免疫侧耳蜗制作石蜡切片。通过免疫组化检测内耳Bcl-2和Bax的表达。结果表明，内耳免疫反应可以诱导细胞凋亡的发生以及Bcl-2和Bax蛋白在其中起了重要的调节作用。     

New strategies for the repair of spinal cord injury

Spinal cord injury （SCI） leads to several neu- rologic damages to patients with sensory and motor func- tion loss. Although some inspiring results were reported in animal, even in non-human primate models, the clinical translation of these research achievements barely move forward for the SCI patients. This review mainly focused on the pathogenesis of the primary and secondary injury, the clinical therapies and the new technologies in spinal cord injury. Moreover, the key scientific issues on this field were discussed in the review. Finally, some suggestions were proposed for the scientific and clinical researches.