目的观察APP-17肽对大鼠多巴胺能神经元变化的影响。方法SD雌性大鼠分4组:对照组、帕金森病(PD)模型组、PD治疗组、APP-17肽预防组。PD以黑质多巴胺能神经元变性和减少为特征。以6-羟多巴胺单侧损毁大鼠脑内多巴胺能系统可建立PD动物模型。通过阿普吗啡诱导旋转,观察行为学改变,并经过高压液相法检测大鼠脑内纹状体和黑质多巴胺及其主要代谢产物水平,可反映多巴胺减少或恢复程度。4周后灌注固定,取大脑组织冰冻切片,作TH免疫组织化学染色。结果(1)行为学检查:APP-17肽预防组及PD治疗组旋转次数明显低于PD模型组。(2)多巴胺及代谢产物含量检测:APP-17肽预防组及PD治疗组代谢产物含量均有明显增加,接近对照组。(3)TH免疫组织化学染色:APP-17肽预防组及PD治疗组多巴胺能神经元数目与正常大鼠相似,明显多于PD模型组。结论APP-17肽对大鼠多巴胺能神经元具有防治和保护作用。

OBJECTIVE To explore condition of pri-mary organ culture of the utricular maculae of the guineapigs inner ear in vitro and establish an experimentalmodel for further research. METHODS The utricularmaculae of the guinea pigs were primarily cultured withtissue culture method in vitro. RESULTS The grow ofthe outgrowth zone of the utricular maculae presentedfibroblasts cell during 10 days. Existing of the support-ing cells and hair cells in sensory epithelium were foundby means of paraffin embedded sections on 10th day.Hair cells were damaged and supporting cells were livedin culture medium with gentamicin for 48 hours bymeans of plastic embedded mount and semi-thinsectioning. CONCLUSION Primary organ culture ofthe utricular maculae of the guinea pig inner ear in vitrois feasible. This organ culture method provide an ex-perimental model for further research on utricularmaculae.

Objective To establish a practical and reliable animal model of spontaneous pain after spinal injury.Methods 30 female Wistar rats were divided into 2 groups: G1 and G2.The injury was induced in L_2 segment of spinal cord using 3 Ncm(G1)and 2 Ncm(G2)by WADE method,respectively.SEP(somatosensory evoked potential) was recorded in L_1 before injury induction and 15 min after injury.The pain threshold of rats in trunk to mechanical gently pressure,gently tactile and rate of forepaws' withdrawing was assessed using von Frey hair just before and after 4 h,8 h,12 h,16 h,24 h injury everyday for about 10 months.Behavior activities of spontaneous pain were also determined everyday.Results Two groups exhibited typical allodynia phenomena.The pain threshold before and after injury has decreased significantly(P<0.05～0.01).The obvious spontaneous pain could be observed.The proportion of serious spontaneous pain(autotomy) in G1 group is 50%,which was near to that of the clinical observations.Conclusion SCI of 3 Ncm in rat model by WADE method has an excellent repeatability.(It's) spontaneous pain symptom is near to that of CP in SCI patients.The animal model will be useful for SCI pain studies.

哺乳动物的神经肌肉接头(NMJ)是被研究最多的突触结构.组织学染色为NMJ形态研究提供了良好途径.本方法总结以往各种染色方案[1～3],针对原有方法的染色背景偏重、神经末梢着染率低、显示不清晰、效果不稳定等缺点,综合Mesulam法和Bielschowsky法[4],摸索出一种改良乙酰胆碱酯酶-硝酸银双染法,染色效果满意,现介绍如下.

通过大鼠脑出血模型血肿周围组织HSP70、NF-κB、Bax和Bcl-2免疫组化研究,探讨脑出血血肿周围组织继发性损害的可能机制.制备大鼠脑出血模型,实验组分成1 h、3 h、12 h、24 h、48 h、72 h及7 d 7个小组;生理盐水对照组分成3 h、24 h及72 h 3个小组,在血肿周边区及同侧皮质区分别取脑组织行HSP70、NF-κB、Bax和Bcl-2抗体免疫组织化学染色,采用HP1000高清晰度彩色病理图像分析系统计数免疫反应阳性细胞数,并与生理盐水对照组进行比较.发现:实验组的HSP70和NF-κB在血肿侧术后72 h表达达高峰(P<0.01);血肿侧各时间点HSP70和NF-κB阳性细胞数与生理盐水对照组相比差异有统计学意义(P<0.01);实验组Bcl-2在术后12 h表达达高峰(P<0.01),而Bax表达在24 h达高峰(P<0.01),二者的血肿侧阳性细胞数与生理盐水对照组相比差异有统计学意义(P<0.01).结果提示:脑出血后血肿周围组织HSP70、NF-κB、Bcl-2和Bax蛋白发生动态变化,参与了血肿周围继发性损害的病理过程.

新近脑内神经营养因子对脑缺血损伤后神经修复与再生的作用与机制研究成为研究的热点.其中脑源性神经营养因子(BDNF)系脑内分布最为广泛的神经营养因子家族成员之一[1、4],与碱性成纤维细胞生长因子(bFGF)一样在中枢神经系统的发育和成熟,以及新生血管形成方面发挥着重要作用.脑缺血后脑内多种神经营养因子均呈现高表达,与缺血后脑组织蛋白表达的变化及分布规律,探讨中药复方脑脉康的脑保护作用及机制.

大量研究表明,脑缺血损伤急性期神经元损伤以坏死为主,凋亡为辅,坏死一般位于缺血核心区,而凋亡多位于周边半暗区,且与梗塞灶的进一步扩大有关.目前多数学者认为,缺血再灌损伤后迟发性神经元死亡(DND)其本质就是凋亡[1].凋亡调控基因bax/bcl-2在脑缺血再灌注损伤后神经细胞凋亡中发挥重要作用[2].本研究旨在通过研究脑缺血再灌注损伤后神经元凋亡及凋亡调控基因bax及bcl-2表达的变化,探讨脑脉康抑制再灌注损伤后神经元凋亡的作用与机制.

目的研究壳聚糖导管-生物活性载体系统诱导神经再生的情况.方法 50只成年雌性Wister大鼠脊髓T7-9节段行脊髓右侧半离断,分别植入含有(40只)或不含有(10只)生物活性载体的壳聚糖导管作桥梁,缝合硬脊膜,恢复脑脊液循环.结果 6-12个月后,在含有生物活性载体的导管中脊髓缺损部位神经再生完成,经过WGA-HRP进行顺行神经束路追踪,发现再生轴索已到达缺损部位的远端,并越过导管远端与宿主的界面进入损伤平面以下脊髓组织,超微结构观察到典型的神经纤维、髓鞘结构和突触.在不含生物活性载体的导管中,再生的轴突很少,且没有轴突穿过导管中点.结论内含生物活性载体的生物材料导管可诱导大鼠脊髓神经轴突的再生.

为观察救脑益智水提液对半乳糖老化小鼠海马神经元的神经元存活信号转导通路和凋亡级联反应中某些蛋白质表达的影响,将昆明种小鼠分为4组(每组24只):正常对照组(C组)、D-半乳糖老化模型组(D组)、大剂量药物治疗组(B组)和小剂量药物治疗组(S组).除C组外,3组小鼠每日皮下注射半乳糖50 mg/kg.其中B组和S组每日分别灌胃救脑益智胶囊水提取液,相当于生药1 g/kg和0.3 g/kg,C组和D组分别灌胃等量生理盐水,共3个月.结果:D组小鼠海马神经元中Akt/PKB(蛋白激酶B)、CREB(cAMP response element binding protein)和Bcl-2的免疫组化阳性细胞数明显减少,而AIF(凋亡诱导因子)和Bax明显增加,PP-1(蛋白磷酸酯酶-1)也明显减少.S组上述改变恢复正常,而B组却无此作用.提示:小剂量救脑益智水提液对半乳糖老化模型小鼠海马神经元的存活信号转导通路和凋亡级联反应的改变具有恢复正常的作用,而大剂量则无此作用.

目的研究人工神经-壳聚糖复合胶原导管修复大鼠坐骨神经15mm缺损的可行性.方法借助人工神经修复10只大鼠坐骨神经缺损15mm,神经缺损7只为对照组,术后2个月、4个月行免疫组化、Osmium染色、Bodian染色、运动终板的特异染色、WGA-HRP神经示踪及大体观察.结果术后2个月再生神经修复了坐骨神经的缺损,实验动物未出现排斥反应及明显的炎症反应.结论人工神经导管对缺损的坐骨神经修复具有良好的桥梁作用和促神经生长的作用.

目的在离体状态下对豚鼠前庭椭圆囊器官进行培养,观察庆大霉素损伤毛细胞后细胞的增殖情况.方法采用白色健康豚鼠36只,体重300-450克,随机分为4组:(1)正常对照组;(2)正常加BrdU(5-溴基脱氧尿核苷BrdU)组;(3)庆大霉素组;(4)庆大霉素加BrdU组.应用体外组织培养、5-溴基脱氧尿核苷(BrdU,3μg/ml)掺入及免疫组织化学染色和Epon树酯包埋半薄切片等方法,在不同时间(2、3、5、15天)光镜观察庆大霉素(2.0mM)损伤后毛细胞及支持细胞增殖情况.结果用庆大霉素48小时后,可见到椭圆囊感觉上皮中的毛细胞溶解破坏.加入BrdU继续培养,BrdU免疫组织化学染色显示了第5和15天被BrdU标记的阳性细胞核,阳性细胞核多位于椭圆囊感觉上皮中基底层,从细胞的形态和分布位置上看为支持细胞;感觉上皮的表层,也可观察到BrdU反应阳性的上皮细胞,细胞核较圆,形态和位置类似毛细胞.结论提示损伤后的椭圆囊感觉上皮支持细胞可以进行细胞的有丝分裂,支持细胞可能是修复毛细胞的前体细胞.

Objective To observe the expression of GFAP and MDR 1 at the resected brain tissues of intractable epileptic patients with varied etiologies.Methods The expression of MDR 1 and GFAP in the surgically resected brain tissues was observed by single and double immunohistochemistry in 22 cases with medically intractable epilepsy.Results The presence of reactive astrocyte seemed to be a common pathological characteristic, though the etiology and clinical performance for each subject were vary. Immunoreactivity of both MDR 1 and GFAP was mainly showed at reactive astrocyte and some areas around micro vessels. Collocalization of MDR 1 and GFAP was also present at most of reactive astrocytes and the areas surrounding micro vessels. Conclusions Reactive astrocyte is one of the most common pathological characteristics in intractable epilepsy. The collocalization of overexpressed MDR 1 and GFAP at some reactive astrocyte is also prominent in brain tissues of intractable epilepsy.

Objective: To research the mechanism of central pain(CP) in spinal cord injury(SCI) and to look for radical curing method . Method: 30 female rats were divided into 2 groups:X 1 and X 2.Injury in L2 segment of spinal cord using 300gcm (X 1)and 200gcm (X 2)by WADE method were conducted, and SEP in L1 segment before and 15 minutes after injury were recorded. Assess pain threshold of rostral flank of trunk to injury level to mechanical gently tactile , gently pressure sense rate of withdrawing forefoot using Von Frey Hair 4,8,12 ,16 ,24 hour and every day after operation for about 10 months. We also observe the behavour of spontaneous pain so on. Result: Two groups exhibited typical acute and chronic allodynia phenomina (Comparison of pain threshold before after operation was found to have significance or much signifiance difference: P 0.05 or 0.01) and spontaneous pain.The incidence of serious spontaneous pain (autotomy)was higher in X 1 than that in X 2,and which is corresponded to that in CP of SCI patients.Conclusion: 300 gcm SCI in rat by WADE method has excellent reappearance and is the closest to CP in SCI in human.Therefore ,it is an excellent animal model of CP in SCI.

目的:研究不同剂量丙泊酚对大鼠脑内血红素氧化酶-2(HO-2)活性的影响.方法:正常SD大鼠24只,随机分为4组:对照组,丙泊酚小、中、大剂量组,每组各6只,依次腹腔注射生理盐水10 mL·kg-1、丙泊酚50,100,200 mg·kg-1.应用免疫组织化学方法,光镜观察丙泊酚麻醉大鼠小脑、端脑和脑桥HO-2阳性神经元的分布.并采用计算机彩色图像分析仪测定各组HO-2阳性细胞数或阳性产物面积.结果:(1)大鼠脑内HO-2阳性神经元分布广泛.(2)腹腔注射丙泊酚各剂量组能明显抑制大鼠脑内HO-2活性.结论:丙泊酚可能抑制大鼠脑内HO-2活性.

Objective To creat a new rodent model of Parkinson's disease (PD) for the investigation of its pathological mechanism. Methods At different survival time after injection of lipopolysaccharide(LPS) in the substantia nigra (SN) of rats, the levels of DA and their metabolites were measured by using HPLC, and the tyrosine hydroxylase (TH) positive neurons and microglia were observed by immunohistochemistry. Results The levels of DA and their metabolites were decreased and the lowest level ( p 0 01) appeared at 14th day after injection of LPS. The number of TH positive neurons was also reduced from the second day after injection, and a large amount of microglial cells were activated with their shapes changed. Conclusions The injected LPS could result in a damage of dopaminergic neurons in the SN.

本文通过锰诱导多巴胺能神经元凋亡及其可能的神经化学机制的研究,进而探讨锰中毒与帕金森病发病的相互关系.分离培养大鼠中脑黑质多巴胺能神经元用不同剂量MnCl2处理后,用荧光染料进行染色,观察了凋亡神经元数量.用腹腔注射及脑内单侧注射MnCl2染毒方法处理大鼠,并采用脑内微透析技术和高效液相色谱-电化学方法(HPLC-ECD)在活体检测了术后不同时间的纹状体细胞外液中DA及其代谢产物DOPAC、HVA以及5-HT的代谢产物5-HIAA等的含量;同时作丙二醛含量和过氧化物歧化酶活性检测.结果发现,凋亡神经元的细胞核缩小、不规则、染色质呈块状深染,凋亡细胞数量随MnCl2剂量升高而增多.MnCl2脑内注射侧与注射对侧相比,术后4、7、10、20 d的DA、DOPAC、HVA和5-HIAA含量均有不同程度的降低.腹腔染毒高、低剂量组20 d后大鼠整体纹状体匀浆的上述指标也明显降低.此外,染毒大鼠纹状体中丙二醛水平随染毒剂量增高而增高,过氧化物歧化酶活性随染毒剂量增高却下降.以上结果表明,锰中毒可能是引起帕金森病发病的原因之一.

目的:用免疫组化法研究人继发青光眼视乳头筛板细胞外基质的改变,探讨眼压增高对筛板的影响.方法:用免疫过氧化酶法(ABC)观察7例晚期继发青光眼筛板中的Ⅳ型胶原蛋白、层连接蛋白的分布.结果:与年龄相匹配正常成人视乳头相比,继发青光眼视乳头筛板前区和筛板部Ⅳ型胶原蛋白和层连接蛋白阳性染色物明显增多,筛板后陷、重叠和融合.结论:青光眼病理过程中眼压增高引起视乳头筛板细胞外基质发生特异性改变,从而使视乳头筛板的生物力学特性发生变化.

锰中毒主要表现为锥体外系神经障碍,并可出现类似帕金森氏综合征的临床表现.近年来由于作为汽油抗爆剂的四乙基铅逐渐被三羰基甲基环戊二烯合锰(methylcyclopentadienyl manganese tricarbonyl,MMT)替代,造成环境中锰的含量逐渐升高,锰对健康的影响更加引起人们关注[1].

用墨汁灌注法、组织化学二氨基联苯胺(DAB)血管染色法及二盐酸联苯胺(BDHC)血管染色法对正常大鼠脑冠状切片进行观察,研究大鼠丘脑、基底节及内囊的微血管构筑,并用Leica显微分光光度计测量光密度值(OD值)来进行定量研究.结果表明,丘脑、基底节及内囊各结构的毛细血管吻合成网,不同神经组织内血管网形态各异,密度差异十分显著.丘脑内毛细血管吻合丰富且复杂.基底节的尾状核及苍白球内血管网形态相似且较密,网孔较小.内囊中血管分布多与内囊脚一致方向排列,管径较粗,网孔较大.各核团的血管密度显著高于神经纤维区的血管密度.