显性脊椎裂胎鼠脊髓神经细胞凋亡及凋亡相关基因表达的意义

目的 观察胎鼠显性脊椎裂脊髓神经细胞凋亡的发生及神经细胞内凋亡相关基因Bcl-2和Bax表达,探讨显性脊椎裂畸形发生过程中脊髓神经细胞凋亡及上Bcl-2和Bax在神经细胞表达的意义.方法 采用原位末端标记法和免疫组化SABC法检测30例脊椎裂胎鼠和14例正常胎鼠脊髓神经细胞凋亡状态及凋亡相关基因Bcl-2、Bax蛋白的表达.神经细胞凋亡用凋亡指数(AI)表示,AI=凋亡细胞数/500×100%.基因Bcl-2、Bax蛋白的表达强度用平均灰度值G表示,蛋白表达越强G值越小.结果 显性脊椎裂胎鼠脊髓神经细胞凋亡数明显增多,正常脊髓神经细胞凋亡指数为0.5%,畸形脊髓神经细胞凋亡指数24%,统计学差异有极显著意义.正常胎鼠脊髓神经细胞Bcl-2蛋白表达强,而脊椎裂胎鼠脊髓神经细胞Bax蛋白表达强,统计学分析二组脊髓神经细胞Bcl-2、Bax蛋白表达差异有显著性意义.结论 在显性脊椎裂发生过程中脊髓神经细胞过度凋亡,提示神经细胞的过度凋亡与显性脊椎裂畸形的发生密切相关.脊椎裂畸形胎鼠脊髓神经细胞Bax蛋白表达强,Bcl-2蛋白表达弱,促进细胞凋亡,推测在脊椎裂畸形发生过程中,Bcl-2和Bax基因对脊髓神经细胞凋亡的发生起重要作用.     

不同胎龄脊髓脊膜膨出胎鼠膀胱和脊髓神经损害的形态学观察

目的 通过不同胚胚胎期脊髓脊膜膨出(MMC)胎鼠膀胱平滑肌的发育和神经支配情况及脊髓神经损害的形态学观察,了解这种损害的时间和空间上的变化及其相互关系.方法 胎鼠分为MMC组和正常对照组,每组再根据胎龄各分3小组,共6小组.用免疫组织化学法检测各组腰骶段脊髓GFAP、VAChT的表达和膀胱α-Actin和neurotubulin-β-Ⅲ的表达；另取各组腰骶段脊髓及膀胱切片作HE染色.Image-Pro Plus 5.1软件分析免疫组织化学照片相关部分的光密度,SPSS 16.0软件对数据进行进行统计分析,以P≤0.05为差异有统计学意义.结果 在胚胎16d,2组胎鼠的膀胱肌层内就已经可以检测出neurotubulin-β-Ⅲ的表达；胚胎16、18和20 d时,MMC组胎鼠膀胱内的neurotubulin-β-Ⅲ表达较相同胚胎期的正常对照组显著降低.在胚胎16、18、20 d,2组的膀胱肌层内都能看到较多的α-Actin的阳性显色；且各胚胎期,2组膀胱内的α-Actin表达差异无统计学意义.在胚胎16d和18d,正常组和MMC组及胚胎20 d正常组的脊髓内,GFAP染色均很少,没有明显的星形胶质细胞显示,在胚胎20 d的MMC组胎鼠的脊髓背侧,可见大量的GFAP阳性的星形胶质细胞显影,其表达显著高于正常组.MMC组各个胚胎期脊髓VAChT的表达都较少,在胚胎20d组的背外侧核仅见少量阳性表达的神经元；而正常组VAChT的表达相对较多,在胚胎20d组的背外侧核可见较多阳性表达的神经元；2组间各胚胎期VAChT表达差异均有统计学意义.结论 MMC胎鼠膀胱平滑肌的分化和发育与正常组差别无统计学意义.MMC胎鼠胚胎16 d,膀胱肌层内可见神经支配,但神经纤维减少,与脊髓内骶副交感神经核和背外侧核的胆碱能神经元减少是同步.MMC胎鼠脊髓损伤后的神经修复过程,星形胶质细胞的出现要晚于胚胎18d.     

脑源性神经生长因子抑制苯丙氨酸诱导的神经元凋亡

目的观察苯丙氨酸是否诱导体外培养神经元凋亡及脑源性神经营养因子（BDNF）对苯丙氨酸诱导的神经元凋亡是否有保护作用。方法胎鼠大脑皮层神经元培养3d后,加入0.9mmol/L苯丙氨酸或同时加入100ng/ml BDNF诱导,原位末端标记法（TUNEL）检测凋亡细胞,Western blotting检测caspase-3的活性。结果神经元在苯丙氨酸诱导18h、30h后,凋亡细胞明显增加（P〈0.01）,而同时加入BDNF后则可减少凋亡的发生（P〈0.05）。并且BDNF抑制苯丙氨酸诱导caspase-3的激活。结论高浓度苯丙氨酸诱导体外培养神经元凋亡依赖cas-pase-3的激活,BDNF能抑制苯丙氨酸诱导的神经元的凋亡。     

骨髓间充质干细胞在先天性脊椎裂胎鼠脊髓中的分化

目的观察大鼠骨髓间充质干细胞(MSC)在先天性脊椎裂胎鼠脊髓中的存活与分化,探寻细胞替代治疗先天性脊椎裂的方法。方法用贴壁培养法进行大鼠MSC原代培养并传代,携带增强型绿色荧光蛋白(EGFP)基因的腺病毒转染第3代骨髓MSC。将转染的MSC注射到先天性脊椎裂胎鼠的脊髓中。母鼠孕20 d时取先天性脊椎裂胎鼠的脊椎,固定脱水后做冷冻切片,免疫荧光方法检测脊髓中EGFP阳性MSC中巢蛋白(Nestin)、胶质纤维酸性蛋白(GFAP)的表达。结果骨髓MSC中CD90阳性细胞占99%,CD44阳性细胞占95%,不表达CD34。腺病毒-EGFP转染骨髓MSC的转染率为61%。致畸组显性脊椎裂胎鼠占64.3%。移植的MSC存在于脊髓表面或进入脊髓中,部分细胞变为长梭形或多角形。免疫荧光法检测结果显示移植到脊髓中的MSC可表达神经干细胞的标志物Nestin和神经胶质细胞的标志物GFAP。结论带有EGFP的MSC经细胞移植后能在胎鼠脊髓内存活,并能分化为神经干细胞和神经胶质细胞。     

显性脊柱裂动物模型膀胱胆碱乙酰转移酶多巴胺β羟化酶和降钙素基因相关肽表达的研究

目的：显性脊柱裂可导致不同程度的膀胱功能障碍,发病机制不清,该研究旨在探讨胆碱乙酰转移梅(CHAT)、多巴胺β羟化酶(DBH)和降钙素基因相关肽(CGRP)的表达在显性脊柱裂所致神经性膀胱功能障碍发生发展中的意义。方法：用维甲酸(RA)致畸Wistar孕鼠,取20d显性脊柱裂胎鼠20只。同时取正常胎鼠20只,行苏木精-伊红和免疫组织化学染色,检测胎鼠膀胱CHAT,DBH和CGRP的表达。结果：在正常胎鼠,膀胱由粘膜、粘膜下、肌层和外膜组成,CHAT,DBH和CGRP广泛分布于膀胱壁各层,以粘膜层、肌层和外膜细胞胞浆着色明显,表达强度光密度值(OD值)分别为398±13,378±14和412±25。显性脊柱裂胎鼠膀胱壁变薄,肌层发育差,CHAT,DBH,CGRP的表达明显减少,OD值分别为156±9,32±6和121±11,差异具有统计学意义(P<0.01)。结论：显性脊柱裂胎鼠膀胱壁肌层发育差,膀胱CHAT,DBH,CGRP的表达明显减少,可能是导致显性脊柱裂胎鼠出生后膀胱功能障碍的机制之一。     

维甲酸致大鼠胚胎神经管畸形的组织形态学观察

目的 利用维甲酸制作先天性神经管畸形的动物模型已获得公认，但对于其导致神经组织错构瘤的发生却未见报道。本实验利用维甲酸口服制作大鼠神经管畸形动物模型，观察其出现神经组织畸胎瘤、脊髓裂开、脊柱裂等畸形形态学特点，探讨其发生机制。方法 利用成熟未孕的Wistar大鼠，在妊娠10dN经胃管注入维甲酸，诱导产生神经管畸形动物模型，分别在妊娠15d、17d、19d时剖宫取出胎鼠，进行实体观察、畸形统计，同时对神经管畸形进行病理形态学研究。对照组仅给予橄榄油，剖宫时间及观察项目同致畸组。结果 维甲酸致畸组共获得胎鼠99只，48．3％(48／99)孕鼠胚胎肉眼观察出现腰骶部显性脊柱裂。在48只显性脊柱裂胎鼠中，分别取不同胎龄(15d、17d、19d)各3只脊髓裂开的畸形胎鼠共9只，做石蜡切片，全部同时见到表现为多个神经管紊乱排列、骶尾部增生肿物，提示神经组织错构瘤的发生。结论 维甲酸可以干扰神经发育，导致脊柱裂、脊髓裂开和神经错构瘤的发生。     

反式维甲酸诱导显性脊柱裂胎鼠骶尾部神经前体细胞凋亡和增殖的变化规律研究

目的观察大鼠胚胎脊柱裂发生早期,细胞凋亡与细胞增殖的变化规律。方法孕鼠随机分为对照组和实验组。胚胎10天时,实验组1次性经胃管注入致畸量反式维甲酸诱导产生脊柱裂动物模型,对照组胃饲等量溶剂,分别在妊娠11、12、13天(E11,E12,E13)时剖宫取胚胎,一部分胚胎固定后进行全胚胎TUNEL染色观察其整体凋亡情况;另一部分胚胎常规制作石蜡切片,采用TUNEL切片染色和免疫荧光染色技术,检测胚胎脊部神经管组织中细胞凋亡和细胞增殖的变化。结果与对照组相比,细胞凋亡于多个发育部位明显增多,集中表现在颅面原基、颅部神经管的背外侧、骶尾部神经管的背部中线。免疫荧光染色显示,与对照组相比,脊柱裂组胚胎畸形发生部位的神经前体细胞的凋亡率升高[E11(2.02±0.52)%与(0.57±0.23)%,E12(3.56±0.33)%与(0.93±0.14)%,E13(3.76±0.37)%与(1.24±0.21)%,P0.001]。而细胞增殖降低[E11(65.17±2.30)%与(81.76±2.17)%,E12(63.97±3.03)%与(76.98±5.14)%,E13(56.86±2.80)%与(73.43±1.99)%,P0.001]。结论反式维甲酸诱导的大鼠脊柱裂胚胎骶尾部神经管中神经前体细胞凋亡增多,而细胞增殖减少,这可能是脊柱裂胚胎神经元发育异常的主要原因之一。     

隐性脊柱裂动物模型脊髓神经递质表达的研究

目的本研究旨在检测丙戊酸诱导的隐性脊柱裂是否存在脊髓神经递质表达的异常。方法用丙戊酸（VPA）致畸孕鼠，取20d隐性脊柱裂胎鼠和正常胎鼠各20只，行免疫组织化学染色，检测胎鼠腰骶段脊髓胆碱乙酰转移酶（ChAT）、神经生长相关蛋白43（GAP43）、多巴胺β羟化酶（DBH）和降钙素基因相关肽（CGRP）的表达。结果 在正常胎鼠脊髓前角运动神经元细胞胞质内可见ChAT、GAP43、DBH和0GRP的阳性表达，ChAT、GAP43、DBH和CGRP表达强度（OD值）分别为452．76±30．86、415．00±30．85、345．21±51．15、221．32±22．42，VPA隐性脊柱裂组可见畸形均发生在腰骶段，脊髓结构无明显异常，ChAT、DBH、CGRP和GAP43的表达均为弱阳性和阴性，ChAT、DBH、CGRP和GAP43表达强度（OD值）分别为55．41±15．88、47．50±15．06、31．73±11．51、21．35±4．29，差异具有统计学意义（P〈0．01）。结论隐性脊柱裂胎鼠脊髓ChAT、DBH、CGRP和GAP43的表达明显减少，可能是导致隐性脊柱裂胎鼠出生后大小便及下肢神经功能障碍的原因之一。     

脊柱裂动物模型制作与胚胎发生机制探讨

目的 建立大鼠隐性脊柱裂和显性脊柱裂畸形动物模型，观察丙戊酸钠和维甲酸诱导大鼠脊柱裂畸形的特点，初步探讨脊柱裂畸形的发生机制。方法 雌性Wistar孕鼠随机分成组：对照组，实验组分成丙戊酸钠三个不同剂量组和维甲酸组。对照组孕鼠不予任何处置，20d胎鼠脊柱染色标本采集图像经计算机软件显微图像分析，测量出每个椎体椎弓两个软骨端距离，目的确定软骨端距离参考值。雌性Wistar大鼠，在确定怀孕的第9d9：00、16：00，按体重计算剂量后于后肢皮下注射20％丙戊酸钠溶液。妊娠第20d剖宫，检查胚胎外观，判定是否有外观可见畸形，然后取脊柱进行骨和软骨双重染色，染色后标本采集图像测量出每个椎体椎弓两个软骨端距离。参照正常胎鼠确定的参考值判定隐性脊柱裂。Wistar大鼠确定怀孕的第10d9：00，胃管灌入维甲酸矿物油混悬液135mg／kg(40mg／m1)，妊娠第20d剖宫，检查胚胎外观判定显性脊柱裂畸形发生情况并记录。结果 丙戊酸钠诱导隐性脊柱裂畸形随药物剂量加大，畸形发生率升高，丙戊酸钠剂量400mg／kg、450mg／kg、500rng／kg~导畸形发生率分别是80％、93％、100％，腰椎对丙戊酸钠的致畸作用最敏感；丙戊酸钠还可以导致头端神经管畸形，未观察到显性脊柱裂的发生。维甲酸135rng／kg诱导胎鼠显性脊柱裂的发生率52．3％，畸形都发生在腰骶段，其他畸形的总发生率高达90．7％。结论 丙戊酸钠抑制软骨细胞增生，影响椎体的软骨发生使椎弓不能闭合导致胎鼠隐性脊柱裂的发生。腰椎对丙戊酸钠的致畸作用最敏感，隐性脊柱裂多发生在腰骶段。过量的维甲酸直接影响神经上皮细胞、神经嵴细胞、原基器官间质细胞的生化过程和尾端未分阶段的中胚层，导致胚胎尾端发育不良综合症复杂畸形出现。丙戊酸钠和维甲酸通过不同机制导致畸形的发生。     

神经管缺损大鼠转化生长因子-β2及其受体的表达

目的建立大鼠脊柱裂动物模型，探讨神经管缺损的产生和转化生长因子-β2（TGF-β2）及其受体（TGFβRI、TGFβRⅡ）的关系。方法维甲酸灌胃建立Wistar大鼠脊柱裂动物模型，用RT-PCR方法检测各组胎鼠脊髓和脑中TGF-β2、TGFβRI及TGFβRⅡ的mRNA表达水平，用免疫组织化学方法检测各组胎鼠脊髓和脑中TGF-β2、TGFβRI以及TGFβRⅡ的蛋白表达水平。结果维甲酸组胎鼠体重明显低于对照组，脊柱裂的发生率为73．1％。脊柱裂胎鼠脊髓组织中TGF-β2的mRNA及蛋白表达水平与对照组相比显著降低，TGFβRI和TGFβRⅡ的mRNA及蛋白水平与对照组相比显著升高。脊柱裂胎鼠脑组织中TGF-β2、TGFβRI和TGFβRII的mRAN及蛋白表达水平与对照组相比无显著差异。结论TGF-β2在神经管的发育中起重要作用，TGF-β2表达的降低与神经管缺损的发生有关。受体TGFβRI及TGFβRⅡ的表达水平可能是反应性的增加，TGF-β2通过它们对神经管的发育起作用。     

Neurotrophic factor expression in newborns with myelomeningocele: preliminary data.

BACKGROUND: Neurotrophic factors play a crucial role in the stimulation of sprouting, synaptic plasticity and reorganization after spinal cord damage. The aim of this study was to investigate the expression of some neurotrophic factors [brain derived neurotrophic factor (BDNF), glial derived neurotrophic factor (GDNF), and nerve growth factor (NGF)] in the cerebrospinal fluid (CSF) of newborns with myelomeningocele (MMC) and to determine their correlations with this malformation. METHODS: To measure the expression of BDNF, GDNF, and NGF, we collected CSF samples of six newborns during the neurosurgical operation to correct the open MMC and of 10 matched controls. Endogenous neurotrophic factor levels were quantified using a two-site immuno-enzymatic assay. The statistical analysis was performed using the Mann-Whitney two-tailed two-sample test. FINDINGS: In the CSF of patients analysis of neurotrophic factor expression showed a significant increase of BDNF, GDNF, and NGF compared to the mean level of the control group (445.8+/-82.3, 86.5+/-2.6, and 59.9+/-6.2 pg/mL, respectively, respect to 10.2+/-5.9, 19.9+/-11.3, and 15.3+/-2.6 pg/mL) (p<0.001). INTERPRETATION: Our study shows an over-expression of neurotrophic factors in the CSF of newborns with MMC. This neurotrophin up-regulation may stimulate axonal sprouting and synaptic reorganization of the damaged neural cells at the site of spinal cord lesion. The neurotrophic factor up-regulation may represent a particularly important biochemical markers of spinal cord damage and might be associated with the severity of spine injury in MMC patients.     

Expression of neurotrophic factors in cerebrospinal fluid and plasma of children with viral and bacterial meningoencephalitis

Aim: To evaluate the expression of neurotrophic factors (nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), glial-derived neurotrophic factor (GDNF)) and their association with the clinical-radiological characteristics and outcome of children with viral and bacterial meningoencephalitis (ME). Methods: Prospective observational clinical study performed on 13 children with ME and 12 controls with non-inflammatory obstructive hydrocephalus. Neurotrophic factor levels in the cerebro-spinal fluid (CSF) and plasma were measured using an immunoenzymatic assay. Results: High levels of NGF and BDNF were demonstrated in all patients, while GDNF levels did not undergo significant variations. NGF expression in the CSF was higher in viral ME than in bacterial ME and was correlated with CSF cellularity (particularly mononuclear cells). BDNF expression in the CSF was higher in bacterial ME than in viral ME and was correlated with CSF cellularity and blood platelet count. No relationships were noted between CSF protein or serum C-reactive protein levels and the expression of neurotrophic factors. Regarding clinical and radiological features, elevated NGF/BDNF levels in the CSF correlated with higher incidence of seizures and prolonged comatose state and with specific radiological lesions. No correlation was found between NGF/BDNF levels and final outcome.

Conclusions: The variations in neurotrophic factor levels may reflect an endogenous attempt at neuroprotection against biochemical and molecular changes during both viral and bacterial ME. The expression of these factors is likely to play a neuro-immunomodulatory or neurosurvival role in ME infections.     

Chop及Grp78基因在先天性脊柱裂大鼠胚胎12天神经管中的异常表达

目的 探讨Chop及Grp78基因在先天性脊柱裂大鼠胚胎12 d神经管及神经上皮干细胞中的表达情况.方法 取畸形大鼠12d胚胎神经管组织,荧光实时定量PCR考察Chop及Grp78基因表达.另取12 d正常大鼠胚胎,分离消化神经管行神经上皮干细胞培养.荧光实时定量PCR考察Chop及Grp78基因表达.结果 先天性脊柱裂大鼠胚胎12 d神经管内Chop基因及Grp78基因表达异常增高.神经上皮干细胞分化为神经元后Chop及Grp78基因的表达上调.结论 Chop及Grp78基因在先天性脊柱裂大鼠胚胎12d神经管中表达异常增高,二者在神经上皮干细胞分化为神经元后表达上调,可能与内质网应激相关的凋亡有关.神经上皮干细胞培养培养体系稳定、高效,可以为胚胎早期发育提供实验模型.     

脊髓拴系松解术后脊柱裂患者的肛管直肠功能研究

目的：与临床对于脊柱裂致膀胱功能障碍的大量研究报道相比,关于脊柱裂患者的肠道功能障碍研究资料非常有限。该文拟研究脊柱裂患者行脊髓拴系松解术后的肛管直肠功能。方法：采用多通道肛管直肠测压技术对因排尿功能障碍而来就诊的21例脊柱裂患者进行肛管直肠功能检测,入选患者皆已于至少2年前行脊髓拴系松解术。结果：脊柱裂患者的最大肛管静息压低于对照组,但二者没有统计学差异(P=0.372)。在嘱脊柱裂患者行最大限度收缩肛门动作时,绝大多数患者肛管压力没有任何升高。在行模拟排便动作时,19例(90.5%)患者表现为盆底功能紊乱型肛管压力变化。直肠肛管抑制反射在所有受检者均存在,诱发该反射所需最小直肠气囊容量在脊柱裂患者组和对照组间差异无显著性(P=0.725);诱发持续性直肠肛管抑制反射所需直肠气囊容量在脊柱裂患者组显著性高于对照组(P<0.001)。直肠感觉阈值在脊柱裂患者显著高于对照组(P<0.0001)。结论：大多数脊柱裂患者不能自主收缩肛门外括约肌,排便时表现为盆底功能紊乱型直肠肛管压力曲线,同时直肠感觉功能也受到严重损害。直肠肛管抑制反射在所有脊柱裂患者均存在,该反射可能受到中枢神经系统的调控。     

骨髓间充质干细胞移植对视网膜病变新生大鼠视网膜细胞凋亡的影响

目的：探讨早产儿视网膜病变（ROP）大鼠玻璃体内注射骨髓间充质干细胞（BMSCs）对视网膜细胞凋亡及视网膜中神经营养素-3（NT-3）、睫状神经营养因子（CNTF）表达的影响。方法：40只7 日龄 Sprague-Dawley新生大鼠随机分为正常对照组、ROP模型组、骨髓间充质干细胞（BMSCs）移植组与磷酸缓冲液组（PBS组）,每组10只。采用高氧法制成 ROP 模型,持续高氧5 d后分别给予BMSCs移植组与PBS组玻璃体内注射BMSCs和PBS。移植7 d后,采用 TUNEL/DAPI、NT-3/DAPI、CNTF/DAPI免疫荧光双标法观察BMSCs移植对视网膜细胞凋亡及NT-3、CNTF表达的影响。结果：移植7 d后,正常对照组视网膜几乎未见TUNEL+ DAPI+细胞,BMSCs移植组视网膜可见少量TUNEL+ DAPI+细胞,显著低于ROP模型组与PBS组（P0.05）;正常对照组视网膜可见极少量的NT-3+DAPI+与CNTF+DAPI+ 细胞,ROP模型组与PBS组大鼠视网膜可见少量NT-3+DAPI+与CNTF+DAPI+ 细胞,较正常对照组增多,但差异无统计学意义（P>0.05）;BMSCs移植组大鼠视网膜NT-3+DAPI+与CNTF+DAPI+ 细胞数均显著多于ROP模型组与PBS组(P0.05）。结论：BMSCs移植可减轻ROP大鼠视网膜细胞的凋亡,其机制可能与其促进ROP大鼠视网膜细胞NT-3与CNTF的表达有关。     

Pathogenesis of an Affected Organ Can Help Us to Understand Normal Organ Development*

Abstract This review discusses the pathogenesis of spina bifida and describes the microscopic appearance of the initial pathological features and their further development in the spinal cords. The embryos and fetuses were derived from pregnant mice given orally 60, 40 or 0 mg/kg of retinoic acid in olive oil on day 8 of gestation and killed one, two or 10 days later. Separation of the primary neural fold from the secondary neural tube was seen in 9- and 10-day-old embryos in which the caudal end of the neuroepithelium was disorganized and the posterior neuropore was open; underneath it, the secondary neural tissue was seen. At term, fetuses showed spina bifida, including myeloschisis, myelocele and diplo-myelia. The cord lesions were located between the third lumbar and second coccygeal levels. Thus, the differentiation and development of the spinal cord proceeds as in chick and human embryos, and the spinal cord caudal to the third lumbar level originates from both neuroepithelium and mesenchyme-like cells while that caudal to the third sacral level originates from mesenchyme-like cells only.     

Nerve growth factor expression correlates with severity and outcome of traumatic brain injury in children.

BACKGROUND: Secondary brain damage after traumatic brain injury (TBI) involves neuro-inflammatory mechanisms, mainly dependent on the intracerebral production of cytokines. In particular, interleukin 1beta (IL-1beta) is associated with neuronal damage, while interleukin 6 (IL-6) exerts a neuroprotective role due to its ability to modulate neurotrophins biosynthesis. However, the relationship between these cytokines and neurotrophins with the severity and outcome of TBI remains still controversial. AIMS: To determine whether the concentration of IL-1beta and IL-6 and neurotrophins (nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), glial-derived neurotrophic factor (GDNF)) in the cerebrospinal fluid (CSF) of children with TBI correlates with the severity of the injury and its neurologic outcome. METHODS: Prospective observational clinical study in a university hospital. CSF samples were collected from 27 children at 2h (Time T1) and 48 h (Time T2) after severe TBI, and from 21 matched controls. Severity of TBI was evaluated by GCS and neurologic outcome by GOS. CSF concentrations of cytokines and neurotrophins were measured by immunoenzymatic assays. RESULTS: Early NGF and IL-1beta concentrations (T1) correlated significantly with the severity of head injury, whereas no correlation was found for IL-6, BDNF, and GDNF. Furthermore, higher NGF and IL-6 and lower IL-1beta expression at T2 were associated with better neurologic outcomes. No significant association was found between BDNF or GDNF expression and neurologic outcome. CONCLUSIONS: NGF concentration in CSF is a useful marker of brain damage following severe TBI and its up-regulation, in the first 48 h after head injury together with lower IL-1beta expression, correlates with a favorable neurologic outcome. Clinical and prognostic information may also be obtained from IL-6 expression.     

Investigation of daytime wetting: when is spinal cord imaging indicated?

BACKGROUND: Most children with daytime wetting have detrusor instability. A minority have neuropathic vesicourethral dysfunction. The commonest cause is spina bifida, which may be closed. Clinical features suggestive of closed spina bifida include cutaneous, neuro-orthopaedic or lumbosacral spine x ray abnormalities, impaired bladder sensation, and incomplete bladder emptying. MRI is the ideal method for detecting spinal cord abnormality. It has been suggested that MRI spine is an unnecessary investigation in children with daytime wetting in the absence of cutaneous, neuro-orthopaedic, or lumbosacral spine x ray abnormalities. Aim: To clarify indications for magnetic resonance imaging (MRI) of the spine in children with voiding dysfunction. METHODS: Retrospective study of children with voiding dysfunction referred from the Guy's Hospital neurourology clinic for MRI spine between April 1998 and April 2000. Clinical notes and results of investigations, including urodynamic studies and MRI spine were reviewed. RESULTS: There were 48 children (median age 9.1 years). Closed spina bifida was detected in five, of whom four had neuropathic vesicourethral dysfunction confirmed by urodynamic studies. Impaired bladder sensation and incomplete bladder emptying were more frequent in these children than in those with normal MRI spine. One child with spinal cord abnormality had no cutaneous, neuro-orthopaedic, or lumbosacral spine x ray abnormalities. CONCLUSION: Spinal cord imaging should be considered in children with daytime wetting when this is associated with impaired bladder sensation or poor bladder emptying, even in the absence of neuro-orthopaedic, cutaneous, or lumbosacral spine x ray abnormalities.     

Successful renal transplantation in children with spina bifida: long term single center experience

We report long-term follow up data on cadaveric renal transplantation for end stage renal failure (ESRF) in spina bifida children. Between February 1989 and July 2001, 12 cadaveric renal transplants were performed in 10 children, eight females and two males. Mean age at transplantation was 13.4 yr (range 9-16). Of the patients, eight were wheelchair bound and two were independently mobile. Before transplantation surgical management of the urological tract included, enterocystoplasty and clean intermittent-self catheterization in five patients and ileal conduit urinary diversion in one. A total of eight patients were on renal replacement therapy before receiving the graft while two underwent preemptive transplantation. The 1- and 5-yr graft survival rates were 81 and 81%, respectively. Four grafts failed--two patients have successfully undergone subsequent transplantation. Causes of graft failure were chronic rejection in two, acute rejection and vascular thrombosis in one and vascular thrombosis in one patient, respectively. Two patients died after graft nephrectomy. At a median follow-up of 4.08 yr (range 1 day to 10.65 yr), eight of the 12 grafts are functioning with median serum creatinine of 123 mmol/L (range 65-169). These data demonstrate the feasibility of cadaveric renal transplantation in patients with spina bifida and ESRF. We currently recommend that patients with spina bifida should not be deprived of the benefits of renal transplantation.