骨髓干细胞移植治疗Duchenne型肌营养不良鼠的实验研究

目的　研究骨髓干细胞移植治疗Duchenne型肌营养不良鼠 (mdx鼠 )的效果。方法　取 4～ 5周龄昆明鼠骨髓干细胞 ,体外培养 3d ,静脉移植到 7Gyγ射线预处理 7～ 8周龄mdx鼠体内。临床监测受体鼠移植物抗宿主病 (GVHD)表现 ,并对移植 12周mdx鼠的运动功能、肌电生理、dystrophin蛋白表达情况进行检测。 结果　 5只 7Gy剂量放疗mdx鼠 ,静脉移植 4 8× 10 6骨髓干细胞 ,3个月后 ,肌电图指标有了部分改善 ;10 %肌纤维表达了dystrophin蛋白。结论　静脉移植同种非同系鼠骨髓干细胞治疗mdx鼠有效 ,显示骨髓干细胞移植治疗DMD有着理想的前景。     

骨髓间质干细胞经静脉移植后在脑缺血大鼠的体内分布与迁移

目的 观察骨髓间质干细胞经静脉移植后在脑缺血大鼠体内的存活、分布及迁移情况，为细胞静脉移植奠定基础。方法密度梯度离心法分离大鼠骨髓间质干细胞(rat bone mescnchymal stem cels，rMSCs)，经体外培养并扩增，线拴法制作大鼠大脑中动脉闭塞(MCAO)脑局灶缺血模型，rMSCs Hoechst33342标记后，通过静脉移植，经过1周、3周、5周后处死大鼠，取脑、心、肺、肾、肝、脾组织切片，在倒置荧光显微镜下观察细胞在体内的存活、分布及迁移情况。结果大鼠脑缺血后，经静脉移植骨髓间质干细胞在脑内能较长时间存活，静脉移植的细胞集中于脑缺血灶周围，而脑缺血对侧及心、肺、肾、肝、脾等组织少有细胞发现。结论rMSCs通过静脉途径进行移植后，能够向损伤部位迁移，与宿主脑组织整合在一起。骨髓间质干细胞经静脉移植用于治疗脑缺血具有可行性。     

不同方法移植骨髓间充质干细胞在体内重要器官的分布及分化

背景：大量实验显示心肌损伤后,趋化或直接心肌内注射的骨髓干细胞可以在体内分化为心肌细胞、血管内皮细胞或血管平滑肌细胞。 目的：观察不同方法移植的骨髓间充质干细胞体内重要器官的分布、生长和分化情况。 方法：选用3月龄日本大耳白兔39只,均制作急性心肌梗死动物模型并随机分为4组,3个组为实验组分别以心肌内直接注射、经冠脉注射、经静脉注射3种不同移植经BrdU标记的自体骨髓间充质干细胞。对照组分为3个亚组,分别经冠脉、静脉和心肌内直接注射等量的生理盐水。4周后处死动物,取心、肺、肝、肾制作组织切片并观察BrdU阳性细胞和BrdU及α-横文肌肌动蛋白双阳性细胞。 结果与结论：①骨髓间充质干细胞移植各实验组在梗死区及其周边均可见到BrdU阳性细胞和BrdU及α-横文肌肌动蛋白双阳性细胞。BrdU阳性细胞间及与宿主心肌细胞间有细胞突起相连,并可见明显的横纹和肌小节。②各实验组新生毛细血管密度较对照组都有增加(P < 0.05),其效果顺序由好到差依次为冠脉注射组、心肌注射组和静脉注射组。③经冠脉和静脉移植组非目的器官肺、肝、肾的标本切片内均发现BrdU阳性细胞表达,且静脉移植组肺内BrdU阳性细胞数多于肝、肾,但均未见有分化趋势和恶性细胞表达。结果提示骨髓间充质干细胞经3种方法移植均能使心肌梗死区及其周边区心肌细胞和毛细血管再生,除心肌注射组外非目的器官都有BrdU阳性细胞表达,但仍以目的器官表达为主。     

不同放疗剂量对肌营养不良鼠骨髓干细胞移植的影响

目的　使用不同剂量γ射线对Duchenne型肌营养不良鼠 (mdx鼠 )照射 ,研究骨髓干细胞移植后mdx鼠缺失蛋白表达的情况。方法　将 18只 7～ 8周mdx鼠随机平均分为A、B、C 3组 ,依次给予 7Gy、7.5Gy、8Gyγ射线预处理 ;3天后行骨髓干细胞移植。移植骨髓干细胞取自 4～ 5周的C5 7BL/6鼠 ,体外培养 3d ,按 1 2× 10 7个细胞 /每只剂量移植给 3组预处理后的mdx鼠。连续观察受体鼠的移植物抗宿主病 (GVHD)程度 ;移植 12周后 ,检测受体鼠体内抗肌萎缩蛋白表达情况。结果　C组GVHD反应较明显 ;A、B、C 3组的股骨肌肌纤维dystrophin蛋白表达率分别为 8%、9%和 13 %。各组间缺失蛋白表达率有显著差异。结论　对 7～ 8周mdx鼠 ,给予不同剂量的γ射线预处理 ,骨髓干细胞移植后 ,8Gyγ射线表达最多 ,7Gy射线照射表达量最少 ;但 8Gyγ射线照射GVHD反应较明显。     

杆状病毒修饰后的脂肪干细胞移植治疗mdx鼠的实验研究

目的利用经杆状病毒基因载体系统进行micro-dystrophin基因修饰后的脂肪干细胞(ADSCs)移植治疗Duchenne型肌营养不良症模型(mdx)鼠,探讨ADSCs移植治疗DMD的安全性及可行性。方法 Mdx鼠60只,分为mdx对照组(30只)和mdx移植组(30只);正常C57小鼠为C57对照组(30只)。体外分离培养小鼠ADSCs,利用杆状病毒基因载体进行micro-dystrophin基因修饰;将基因修饰后的ADSCs经尾静脉移植到mdx鼠体内。于移植后检测mdx鼠的运动功能(采用主动牵引实验和被动转棒实验)、血清CK水平、肌肉病理改变以及肌肉micro-dystrophin表达水平。结果经micro-dystrophin基因修饰的ADSCs移植后,能够重建mdx鼠的micro-dystrophin表达,一定程度上减轻并逆转肌肉的病理损害,进而降低血清CK水平,mdx鼠整体运动功能也有一定改善。结论 ADSCs治疗mdx鼠后,可部分重建模型鼠的dystrophin表达,改善肌肉的病理损害,表明ADSCs是有希望治愈DMD的方法之一。     

骨髓移植治疗Duchenne型肌营养不良症模型鼠后dystrophin相关蛋白表达

目的观察骨髓移植治疗Duchenne型肌营养不良症(DMD)模型鼠—mdx鼠后，β-dystroglycan和α-sarcoglycan在肌膜上的表达情况。方法以正常C57鼠作为供体，以mdx鼠作为受体进行骨髓移植，在移植后2、4、6个月分别进行mdx鼠骨骼肌的β-dystroglycan和α-sarcoglycan免疫荧光染色，计算阳性细胞的荧光积分光密度，与C57鼠及未移植的mdx鼠进行比较。结果在骨髓移植后2、4、6个月mdx鼠骨骼肌肌膜上β-dystroglycan和α-sarcoglycan的表达量均有随时间的延长逐渐增加的趋势．至移植后6个月时，二者的表达量明显高于未治疗的mdx鼠。结论骨髓移植可使mdx鼠的dystrophin相关蛋白在病损骨骼肌细胞膜上表达增加．其对维持肌膜的稳定性、促进肌肉的恢复有重要意义。     

Duchenne肌营养不良模型鼠骨髓移植后dystrophin的表达

目的　观察Duchenne型肌营养不良模型鼠 (mdx鼠 )不同成分骨髓细胞移植后dystrophin的表达。方法　用体外培养的C57BL雄鼠的骨髓细胞、骨髓悬浮细胞、骨髓基质细胞分别经鼠尾静脉注射到放疗后的mdx鼠体内 ,动态观察受体雌性mdx鼠骨骼肌dystrophin的表达 ,并取受体mdx雌鼠血 ,用聚合酶链反应进行Sry基因检测。结果　骨髓细胞、悬浮细胞、基质细胞移植后 1～ 2个月 ,较少肌纤维表达dystrophin(<1 % ) ,3～ 4个月分别约有 7%、6 %、4 %的肌纤维表达dystrophin。Sry基因检测均扩增出受体雌鼠Y染色体上 449bp的DNA片段 ,提示供体细胞在受体内存活。结论　骨髓细胞、悬浮细胞、基质细胞分别移植到mdx鼠体内后 ,mdx鼠骨骼肌均有dystrophin的表达     

经静脉途径移植骨髓间充质干细胞在慢性心肌梗死大鼠体内的分布

背景：目前的研究中，大多针对急性心肌梗死进行细胞移植研究，而对于慢性心肌梗死后经静脉移植干细胞的研究尚少见，尤其是在移植后多个时间点动态监测移植的干细胞在体内分布及存活情况尚无报道。 目的：观察经静脉途径移植骨髓间充质干细胞后在慢性心肌梗死模型大鼠体内的分布情况。 方法：从雄性SD大鼠获取培养骨髓间充质干细胞，将18只雌性SD大鼠制备成心肌梗死3周后随机均分为2组，实验组：用胰岛素针抽取加入PBS的含有5×106骨髓间充质干细胞的混悬液300 μL，并将其缓慢注入股静脉；对照组：注入同等体积的PBS。另选9只雌性SD大鼠仅开胸，不结扎冠状动脉，作为假手术组，并将含有5×106骨髓间充质干细胞的混悬液300 μL通过股静脉注入体内。骨髓间充质干细胞移植24 h，2周及1个月后，各组大鼠体内的细胞分布情况应用实时荧光定量PCR技术进行检测。 结果与结论：实验组与假手术组大鼠在细胞移植后1 d，肺组织、肝脏、脾脏中细胞分布差异无显著性意义(P > 0.05)，而实验组心脏中细胞分布明显多于假手术组(P < 0.01)；移植后2周，两组肺组织中细胞数目均急剧减少(P < 0.05)，肝脏、脾脏、心脏中细胞分布差异无显著性意义(P > 0.05)；移植4周后两组相比，肺组织、肝脏、脾脏中细胞分布差异亦无显著性意义(P > 0.05)，而假手术组心脏中未检测到细胞。移植4周后，各组织中细胞分布均明显减少。在对照组中，各时间点均未检测到SRY基因。提示慢性心肌梗死经静脉移植骨髓间充质干细胞后，早期大量细胞滞留于肺组织，且细胞数目随时间延长锐减。此外，心脏，脾脏，肝脏中仍可有少量细胞分布。     

mdx鼠骨髓间充质干细胞体外分离培养及向肌细胞的诱导分化

背景：自体干细胞移植治疗可以克服异体干细胞移植治疗的局限性,但目前对于进行性肌营养不良模型mdx鼠骨髓间充质干细胞生物学特性及肌分化潜能研究未见报道。 目的：拟建立体外分离培养mdx鼠骨髓间充质干细胞的方法,并观察其成肌分化能力。 设计、时间及地点：细胞学体外观察,于2006-01/07在中山大学附属第一医院神经病学实验室完成。 材料：4~6周龄纯系雄性mdx鼠10只,购自美国Jackson实验室,饲养于中山大学北校区动物实验中心。碱性成纤维细胞生长因子和兔抗鼠MHC抗体为Chemicon公司产品。 方法：全骨髓法体外分离mdx鼠骨髓间充质干细胞,差速贴壁法纯化扩增,胰酶+EDTA消化传代。取传至第3代的细胞,按1×104/孔接种于24孔板,置于含两性霉素B、碱性成纤维细胞生长因子、胎牛血清的DMEM培养基中向肌细胞方向诱导分化,2周后更换为不含两性霉素B、但添加马血清的DMEM培养基继续培养2周。 主要观察指标：鼠骨髓间充质干细胞形态、表面标志、生长曲线及成肌诱导分化。 结果：原代培养24 h后,鼠骨髓间充质干细胞约80%贴壁生长,第3代细胞多呈梭形,形态比较均一。鼠骨髓间充质干细胞表达CD29和CD44等间质类细胞的表面分子,不表达CD11B和CD45等淋巴造血细胞的表面分子。接种后第一两天细胞处于潜伏期,第3天细胞增殖旺盛,进入对数生长期,至第4天细胞数增长一倍,第5天进入平台期。两性霉素B诱导后细胞有少量死亡,停用后更换马血清促进细胞成肌,7~10 d可见有肌管样细胞形成,免疫荧光染色可见肌管样细胞呈MHC阳性,DAPI复染后可见肌细胞内有多个细胞核。 结论：在两性霉素B与马血清依次诱导条件下,体外可成功分离培养基因缺陷mdx鼠骨髓间充质干细胞,并证明其具有成肌分化潜能。     

骨髓间充质干细胞移植多器官功能不全综合征兔心、肝、肺器官中凋亡因子Apaf-1的表达*

背景：研究证明多器官功能不全综合征的发病及其器官组织损伤与细胞凋亡和坏死有关,而近年来骨髓间充质干细胞作为种子细胞在修复领域已初现成效。 目的：观察骨髓间充质干细胞移植对多器官功能不全综合征兔受累脏器组织细胞凋亡蛋白酶活化因子1表达的影响。 方法：实验组建立两次打击多器官功能不全综合征动物模型,进行兔自体骨髓间充质干细胞体外培养、扩增、鉴定,慢病毒转基因标记GFP,经股动脉移植,并设立模型对照组及正常组。组织切片观察骨髓间充质干细胞在多器官功能不全综合征动物模型宿主内的存活情况,RT-PCR测定骨髓间充质干细胞移植前后凋亡因子Apaf-1在心、肝、肺重要脏器中的表达。 结果与结论：实验组兔肝、肺脏器组织内有慢病毒转基因标记GFP的骨髓间充质干细胞存在。RT-PCR结果显示正常组织心、肝、肺中均有Apaf-1 mRNA表达,且模型对照组心、脾、肾Apaf-1 mRNA表达较正常组明显增加,而实验组兔心、脾、肾组织中Apaf-1 mRNA表达下降。说明骨髓间充质干细胞具有良好的生物学活性,移植后的骨髓间充质干细胞能够定植在多器官功能不全综合征兔的受损脏器组织内,并下调Apaf-1 mRNA的表达,提示骨髓间充质干细胞移植有望为多器官功能不全综合征提供新的治疗方法。     

TNF-alpha antibody infusion impairs survival of stroke-generated neuroblasts in adult rat brain

Stroke induced by 2 h middle cerebral artery occlusion triggers increased striatal and hippocampal neurogenesis in adult rats. We investigated the effect of tumor necrosis factor-alpha (TNF-alpha) inhibition on the survival of the new neurons. The mitotic marker BrdU was given on days 5 to 7, and TNF-alpha antibody or control protein was infused into the lateral ventricle of the ischemic hemisphere from day 8 to 14 after stroke. At the end of infusions, the TNF-alpha antibody-treated rats showed markedly fewer new striatal and hippocampal neurons, as compared to animals given control protein. The present findings suggest that TNF-alpha, probably acting via its receptor TNFR2, can promote the survival of stroke-generated hippocampal and striatal neurons.     

Enhancement of susceptibility of adult mouse brain to cytomegalovirus infection by infusion of epidermal growth factor

Neural precursor cells, including neural stem and progenitor cells, in the subventricular zone (SVZ) are the main targets for cytomegalovirus (CMV) infection in developing brains. The neural precursor cells in the SVZ of the adult brain have been reported to respond by proliferating after infusion with epidermal growth factor (EGF). Here we report the susceptibility of the precursor cells in the adult mouse brain to murine CMV (MCMV) infection. Adult mouse brains from 10-, 25-, and 70-week-old (W) mice were infused with either phosphate-buffered saline or EGF into the brain for 3 days, and then intracerebrally infected with MCMV for 5 days. The susceptibility of the adult brains to MCMV was significantly increased by infusion of EGF in terms of viral titers and viral antigen-positive cells. The susceptibility of the young adult brain from 10-week-old mice to MCMV was higher than that of the adult brains from 25-week-old or 70-week-old mice. Both the ependymal and the SVZ cells were susceptible to MCMV infection. The number of virus-infected cells in the SVZ was significantly increased by infusion of EGF, whereas the number of infected ependymal cells was not significantly increased. Among the virus-infected cells in the SVZ, 73% were positive for nestin, 87% were positive for Musashi, 86% were positive for GFAP, and 96% were positive for PCNA. These results indicate that the susceptibility of the adult brain to MCMV is correlated with the proliferative ability of the neural precursor cells in the SVZ of the adult brain.     

Chronic infusion of CDNF prevents 6-OHDA-induced deficits in a rat model of Parkinson's disease

The visual system is widely used as a model in which to study neurotrauma of the central nervous system and to assess the effects of experimental therapies. Adult mammalian retinal ganglion cell axons do not normally regenerate their axons for long distances following injury. Trauma to the visual system, particularly damage to the optic nerve or central visual tracts, causes loss of electrical communication between the retina and visual processing areas in the brain. After optic nerve crush or transection, axons degenerate and retinal ganglion cells (RGCs) are lost over a period of days. To promote and maintain axonal growth and connectivity, strategies must be developed to limit RGC death and provide regenerating axons with permissive substrates and a sustainable growth milieu that will ultimately provide long term visual function. This review explores the role olfactory glia can play in this repair. We describe the isolation of these cells from the olfactory system, transplantation to the brain, gene therapy and the possible benefits that these cells may have over other cellular therapies to initiate repair, in particular the stimulation of axonal regeneration in visual pathways. This article is part of a Special Issue entitled: Understanding olfactory ensheathing glia and their prospect for nervous system repair.     

Enhanced acetylcholinesterase staining in hippocampal area CA3 after lesion of granule cells by infusion of colchicine

Unilateral infusion of colchicine into the lateral ventricle produced relatively selective destruction of dentate granule cells in the ipsilateral dorsal hippocampal formation of the rat. Timms silver sulfide stain is markedly reduced in the mossy fiber layer on the colchicine treated side but is normal contralaterally. After colchicine treatment, an increase in acetylcholinesterase staining is apparent in the apical dendritic zone of CA3 pyramidal cells. This enhanced staining is localized in the proximal apical dendritic layer of CA3, a region normally occupied by the mossy fiber terminals of dentate granule cells. These results suggest that cholinergic fibers proliferate in CA3 after granule cell lesion and may participate in reinnervation of the denervated area.     

EGF infusion stimulates the proliferation and migration of embryonic progenitor cells transplanted in the adult rat striatum

Immature progenitor cells (generated by in vitro propagation) may provide a useful alternative to primary cells (from dissected embryonic tissue) for transplantation if their migratory and proliferative and differentiation properties can be controlled and directed in vivo. In this study E15 murine EGF-responsive progenitor cells were transplanted to the striatum of adult rats. Simultaneously, these animals received continuous infusion of either epidermal growth factor (EGF) or vehicle, to the lateral ventricle, for 8 days. In animals that received EGF, the transplanted progenitors migrated toward the lateral ventricle and proliferated, as evidenced by bromodeoxyuridine incorporation. Progenitor cells transplanted to rats that received vehicle infusions showed neither of these responses. In all animals, transplanted progenitors expressed an immature astrocyte or oligodendrocyte phenotype, the majority of cells being astrocytes. We conclude that EGF stimulates the migration and proliferation of murine progenitor cells in vivo, either directly or indirectly, but does not influence their phenotypic differentiation.     

Intraventricular infusion of N-methyl-d-aspartate

Summary The purpose of this study was to document the early cerebrovascular consequences of excessive N-methyl-d-aspartate (NMDA) receptor activation. Five microliters of NMDA (100 nmol/l) or vehicle was infused over a 15-min period into the lateral ventricle of adult rats. The protein tracer horseradisch peroxidase (HRP) was injected intravenously for blood-brain bartier (BBB) studies. The intraventricular infusion of vehicle (n=5) caused no alterations in arterial blood pressure or microvascular damage away from the intraventicular probe tract. In contrast, NMDA infusion (n=8) led to a gradual increase in arterial blood pressure (mean 36 mm Hg). Multifocal regions of HRP extravasation were observed bilaterally throughout the neuraxis following NMDA infusion. Sites of BBB disruption and hemorrhage included brain regions bordering ventricular spaces. In addition, isolated foci of protein extravasation were commonly detected in the cerebral cortex, thalamus, basal forebrain, septum and cerebellum. Pretreatment with the noncompetitive NMDA antagonist MK-801 (2 mg/kg) substantially reduced the BBB responses to NMDA. However, microvascular abnormalities were seen in NMDA-infused rats where blood pressure elevations were inhibited by blood removal. In addition to neurons, cerebral blood vessels are also acutely affected by NMDA receptor activation. Blockage of NMDA receptor channels following brain injury may potentially provide protection by attenuating BBB breakdown and subsequent brain edema.Supported by USPHS Grants NS-05820, NS-30291 and NS-27127 and by the American Heart Association Grant-in-Aid 90-1133, with funds contributed by the Flordia Affiliate.     

Continuous subcutaneous infusion of pramipexole protects against lipopolysaccharide-induced dopaminergic cell death without affecting the inflammatory response

The D2/D3 dopamine receptor agonist pramipexole, protects against toxin-induced dopaminergic neuronal destruction but its mechanism of action is unknown. Inflammation following glial cell activation contributes to cell death in Parkinson's disease and we now report on the effects of acute or chronic administration of pramipexole on lipopolysaccharide (LPS) induced inflammation and nigral dopaminergic cell death in the rat. At 48 h and 30 days following supranigral administration of LPS, approximately 70% of tyrosine hydroxylase (TH) immunoreactive (-ir) cells in substantia nigra had degenerated with a corresponding loss of TH-ir terminals in the striatum. In rats acutely treated with pramipexole (2 × 1 mg/kg; s.c.) 48 h following LPS application, there was no difference in the number of TH-ir cells or terminals compared to LPS-treated rats receiving vehicle. However, the continuous subcutaneous infusion of pramipexole for 7 days prior to LPS and 21 days subsequently, produced a marked preservation of both TH-ir cells and terminals. At 48 h or 30 days, LPS induced an up-regulation of ubiquitin-ir within the nigral TH-ir neurones, which was reduced by pramipexole treatment. Thirty days following supranigral LPS administration (9 days after the end of infusion), (+)-amphetamine (5 mg/kg, i.p.) caused robust ipsiversive rotation. In rats treated with LPS but receiving continuous subcutaneous administration of pramipexole, (+)-amphetamine-induced rotation was markedly reduced. LPS-induced increase in the levels of inflammatory markers, were not affected by either acute administration or continuous infusion of pramipexole. Continuous infusion of pramipexole protected dopaminergic neurones against inflammation induced degeneration but without modification of the inflammatory response.     

Intraventricular infusion of N-methyl-D-aspartate

This study documents the ultrastructural features of acute neuronal injury following N-methyl-D-aspartate (NMDA) receptor activation. NMDA (100 nmol/microliters) or vehicle was infused over a 15-min period into the lateral ventricle of adult rats. After perfusion fixation, specimens demonstrating normal and abnormal patterns of vascular permeability to horseradish peroxidase were sampled for ultrastructural analysis. In NMDA-infused rats, brain regions exhibiting protein extravasation contained swollen dendritic profiles and abnormal neuronal perikarya. Although periventricular regions were most severely affected, parenchymal abnormalities were also detected in the cerebral cortex, septum, striatum, thalamus, hypothalamus and cerebellum. Mildly affected dendrites contained dark compact mitochondria, while in severely swollen dendrites mitochondria were enlarged with ruptured cristae. Focal sites of plasma membrane disruption were also detected within swollen dendrites. Swollen neurons commonly displayed peripheral pallor and increased numbers of cytoplasmic vacuoles. Other neurons appeared dark and shrunken, some containing disrupted mitochondria and pyknotic nuclei. Pretreatment with the NMDA antagonist MK-801 (2 mg/kg) attenuated the neuronal and dendritic alterations. In conditions where cerebrospinal fluid levels of glutamate are abnormally elevated, excessive NMDA receptor activation may lead to early vascular and neuronal complications which could work in concert to promote brain injury.     

Undesired effects of a combinatorial treatment for spinal cord injury--transplantation of olfactory ensheathing cells and BDNF infusion to the red nucleus

Transplantations of olfactory ensheathing cells (OECs) have been reported to promote axonal regeneration and functional recovery after spinal cord injury, but have demonstrated limited growth promotion of rat rubrospinal axons after a cervical dorsolateral funiculus crush. Rubrospinal neurons undergo massive atrophy after cervical axotomy and show only transient expression of regeneration-associated genes. Cell body treatment with brain-derived neurotrophic factor (BDNF) prevents this atrophy, stimulates regeneration-associated gene expression and promotes regeneration of rubrospinal axons into peripheral nerve transplants. Here, we hypothesized that the failure of rubrospinal axons to regenerate through a bridge of OEC transplants was due to this weak intrinsic cell body response. Hence, we combined BDNF treatment of rubrospinal neurons with transplantation of highly enriched OECs derived from the nasal mucosa and assessed axonal regeneration as well as behavioral changes after a cervical dorsolateral funiculus crush. Each treatment alone as well as their combination prevented the dieback of the rubrospinal axons, but none of them promoted rubrospinal regeneration beyond the lesion/transplantation site. Motor performance in a food-pellet reaching test and forelimb usage during vertical exploration (cylinder test) were more impaired after combining transplantation of OECs with BDNF treatment. This impaired motor performance correlated with lowered sensory thresholds in animals receiving the combinatorial therapy - which were not seen with each treatment alone. Only this combinatorial treatment group showed enhanced sprouting of calcitonin gene-related peptide-positive axons rostral to the lesion site. Hence, some combinatorial treatments, such as OECs with BDNF, may have undesired effects in the injured spinal cord.     

丙泊酚复合瑞芬太尼靶控输注在神经外科手术中的应用

目的探讨丙泊酚复合瑞芬太尼靶控输注全静脉麻醉在神经外科手术中应用的临床意义。方法对66例神经外科择期手术病人采用丙泊酚复合瑞芬太尼靶控输注全静脉麻醉。丙泊酚、瑞芬太尼靶浓度分别为2￣4m g/L和2￣5μg/L,间断追加维库溴胺。记录围麻醉期血流动力学、麻醉药用量以及麻醉后恢复情况。结果麻醉诱导后病人收缩压、舒张压均显著性降低(P<0.05),心率减慢(P<0.05),气管插管、切皮前后无明显改变,手术结束后睁眼时心率明显增快(P<0.05),麻醉恢复时病人苏醒较快,自觉舒适,无呼吸再抑制现象。结论丙泊酚复合瑞芬太尼靶控输注,麻醉诱导迅速,维持平稳,停药后清醒快,对气管导管耐受性好,适用于神经外科手术。