神经生长因子诱导后的交感神经元样PC12细胞与新生大鼠心肌细胞共培养的实验研究

探讨神经生长因子(NGF)诱导的交感神经元样PC12细胞作为体外再造心肌神经支配研究模型的可行性。用含0.04?TA的0.25%胰酶分离新生大鼠原代心肌细胞,然后与NGF诱导的交感神经元样PC12细胞共培养,通过光学显微镜观察、常规H.E.染色、激光共聚焦显微镜和扫描电镜观察等对其进行评价。结果表明:在二维共培养模型中,NGF诱导的交感神经元样PC12细胞长出神经突起,突起及其上的膨体能够到达跳动的心肌细胞表面,神经突起随心肌细胞一起跳动。采用神经元样PC12细胞作为体外再造心肌神经支配研究模型是可行的,神经细胞与心肌细胞可能存在支配关系。     

血管周肾上腺素能神经密度与管壁组成成分的关系

本文应用组织化学技术和电镜方法对家兔、豚鼠和大鼠血管周的肾上腺素能神经密度与管壁组成成分的关系进行了观察。实验结果证明,血管周神经分布于外膜层,中膜内未见有神经分布,肌性动脉(以肠系膜动脉为代表)较弹性动脉(以颈总动脉为代表)的血管周神经密度和含膨体数都较高。神经肌肉间隔近(0.05—3微米),最近者神经与肌肉间除基板外无其它组织成分。弹性动脉神经肌肉间隔较远(1—12微米),神经肌肉间隔以外弹力膜、成纤维细胞和板层状的结缔组织。股动脉和肾动脉周的神经分布特点介于上二者之间。静脉较相应动脉神经分布稀疏。但也存在部位的特殊性和种属差异性。如脐动脉虽属肌性动脉,但动脉周并无神经分布,豚鼠肾动脉周的神经密度远较兔及大鼠稀疏。作者认为血管周的神经密度与血管壁中平滑肌的含量有关。本文并对肌性动脉周神经分布致密的原因,不同类型动脉的神经分布特点与生理功能的关系进行了讨论。     

重组神经生长因子受体基因在受体缺乏的神经母细胞瘤细胞系中的表达

神经母细胞瘤是儿童期的常见恶性肿瘤之一。临床上,半数以上神经母细胞瘤有明显的N-myc癌基因扩增。神经生长因子(NGF)可使某些神经母细胞瘤细胞分化成神经元样的细胞。但对有N-myc扩增的神经母细胞瘤则可能因NGF受体缺乏而无明显促分化反应。本研究运用重组DNA技术将人NGF受体基因重组到有N-myc扩增,且NGF受体表达很低的神经母细胞瘤系(IMR-32)中,经克隆化培养、筛选,建立了稳定的细胞系—IMR-32/NGFR和对照细胞系IMR-32/NEO。经用抗NGFR的单克隆抗体检测用Flow cytometry技术证实IMR-32/NGFR系中有明显的NGF受体表达,而其母系IMR-32和空病毒对照系IMR-32/NEO则无表达迹象,说明NGF受体表达在IMR-32/NGFR系中是特异的,并能同抗NGFR单克隆抗体特异结合。用Northern B10ting技术亦测得IMR-32/NGFR中NGFR的mRN A明显表达。而其母系IMR-32和对照系IMR-32/NEO则无明显表达。这说明IMR-32/NGFR系中NGFR在mRNA水平上亦是特异的。N-myc和K-ras癌基因在IMR-32、IMR-32/NEO、IMR-32/NGFR三系中无明显变化。在NGF处理后,形态上三系均无明显的分化迹象。但IMR-32/NGFR在神经原纤维轻链的表达上轻度增高。c-fos癌基因的表达见于所有三个细胞系,IMR-32/NGFR略强些。这些说明,在恢复了NGFR基因和表达之后,对N-myc和K-ra     

脑内和肌内接种小鼠胶质母细胞瘤株后神经末梢的观察

本实验应用Nonidez及Glees二种镀银法,对诱发的小鼠胶质母细胞瘤株(G 422)进行了观察。脑内及肌内接种胶质母细胞瘤后,在肿瘤边缘可见大小不等的神经束,伴随或不伴随血管伸入瘤内。有的在血管周围间隙形成血管周围神经丛。这些神经纤维与肿瘤周围宿主的脑组织、皮下、毛囊和肌肉间隙的神经纤维相联系。因此我们推测肿瘤内的神经是由肿瘤周围宿主的器官组织的神经延伸来的。肿瘤边缘的神经纤维的数量多于核心区,走行于肿瘤的间质或实质,沿途不断分支,终末分布到肿瘤细胞的表面。我们观察到球形、游离分叉状、梭形、环形、树枝状、杵状及丛刷状等类型的神经末梢。以上观察表明,恶性肿瘤——小鼠胶质母细胞瘤是受神经支配的。     

神经生长颗粒对大鼠腓总神经横断损伤的修复作用

目的 研究神经生长颗粒(NGG)对大鼠腓总神经横断损伤的修复作用.方法 SD大鼠50只,随机分为NGG高、中、低剂量组(剂量分别为5.2g生药/kg、2.6g生药/kg、1.3g生药/kg)、弥可保组(剂最为625μg/kg)、空白对照组.行大鼠腓总神经横断缝合术,术后每日灌胃给药.于术后2周、3周、4周行足迹实验,测定展趾功能,术后4周行电生理检测,测定复合肌动作电位和神经干动作电位检测,组织形态学分析,测定再生有髓神经纤维数、髓鞘厚度和胫前肌肌纤维截面积,观察NGG对大鼠腓总神经损伤的修复作用.结果 与空白对照组相比,NGG组展趾功能、复合肌动作电位和神经干动作电位波幅及恢复率、再生有髓神经纤维计数、髓鞘厚度及胫前肌横截面积均显著增高,且呈剂量依赖的量效关系.结论 NGG有利于轴突生长和髓鞘形成,可以促进大鼠损伤神经修复和神经功能的恢复.     

神经脉冲波传播形态的研究

Hodgkin-Huxley模型（H—H模型）是研究神经电生理不可或缺的数学依据。但是，到目前为止，对H—H神经元模型的分析缺少解析研究。本文对经典的H-H模型进行具体分析，得到简化H—H模型以及Nagumo方程，利用齐次平衡方法首次求出简化H-H方程和Nagumo方程的孤波解。研究表明：神经冲动可以孤波的模式传播。     

Reticular and plexiform perineurioma: clinicopathological and immunohistochemical analysis of two cases and review of perineurial neoplasms of skin and soft tissues

Perineurioma represents a recently described and relatively rare neoplasm in the spectrum of benign peripheral nerve sheath tumours composed of perineurial cells staining immunohistochemically positive for epithelial membrane antigen (EMA). In addition to intraneural, extraneural and sclerosing perineurioma, rare variants of perineurioma may occur, and their knowledge is important in the differential diagnosis of mesenchymal tumours of different lines of differentiation and clinically more aggressive neoplasms. We present a case of deep-seated reticular perineurioma arising on the upper arm of a 34-year-old female and a case of a dermal plexiform perineurioma arising on the lower lip of a 60-year-old female. The diagnosis was confirmed in both cases immunohistochemically; neoplastic cells stained positively for EMA and for the newly described perineurial markers, claudin-1 and glut-1. The morphological spectrum and the differential diagnosis of perineurial neoplasms of skin and soft tissues are discussed.     

Inhibition of voltage-dependent Na+ and K+ currents by forskolin in nodes of Ranvier

The effect of forskolin on voltage-activated Na⁺ and K⁺ currents in nodes of Ranvier from the toad, Bufo marinus, has been examined using the vaseline-gap voltageclamp technique. Peak Na⁺ currents (I _Na) were reduced by 35% and the rate of decline of Na⁺ current during continuous depolarization was accelerated following treatment with 450 M forskolin. However, the voltage-dependence of steady-state inactivation as well as the rate of recovery from fast inactivation remained unchanged. Upon repetitive depolarization at 1–10 Hz, a further inhibition of I _Na (60%) was observed. This use-dependent or phasic inhibition recovers slowly at -80 mV ( 13 s) and had a voltage-dependence like that of activation of the Na conductance. Near maximal steady-state phasic inhibition occurred with depolarizing pulse durations of only 4 ms, consistent with a direct involvement of the open Na⁺ channel in the blocking process. Inhibition of the delayed K⁺ current (I _K) was characterized by a concentration-dependent reduction in steady-state current amplitude (IC₅₀ 80 M) and a concentration-independent acceleration of current inactivation. A similar inhibition of I _K was obtained with 1,9-dideoxyforskolin, a homolog which does not activate adenylate cyclase (AC). The results suggest that the inhibition of I _K and perhaps I _Na follows directly from drug binding and is not a consequence of AC activation.     

Effects of nerve growth factor and glucocorticoid on cultured human pheochromocytoma cells

Primary cell cultures of two human pheochromocytomas (PC) that were associated with high serum levels of adrenaline and noradrenaline were developed to study the effects of nerve growth factor (NGF) and dexamethasone on the morphology and function of PC cells in vitro. By phase-contrast microscopy, cultured cells were small and hyperchromatic on the first day of culture; neurite-like processes that extended to other cells developed several days later and were maintained for more than 3 months. NGF (100ng/ml), dexamethasone (10^–5M), or NGF + dexamethasone were added to the culture media 2 weeks after the cultured cells had stabilized. Catecholamine concentrations in the medium were maintained at higher levels after addition of NGF, dexamethasone, or NGF + dexamethasone as compared to control cells. In the presence of NGF, extension of neurite-like processes was clearly accelerated, while high levels of dexamethasone inhibited growth of processes. These in vitro studies showed that the addition of NGF or the removal of dexamethasone induces differentiation of adrenal neurons present in pheochromocytomas, suggesting that adrenocortical steroid hormones influence the morphological control of adrenal medullary cells.     

神经生长因子对穹窿海马伞切割后海马自体神经干细胞增殖和向神经元分化的影响

目的 探讨切割穹窿海马伞及脑室给予神经生长因子(NGF)后,对大鼠海马自体神经干细胞增殖和分化为神经元的影响.方法 12只SD大鼠,随机分成给药组和对照组,每组6只,切割右侧穹窿海马伞,两组切割后即时及第2d、4d向侧脑室分别注射NGF和人工脑脊液, 并在术后第3～7d每日腹腔注射2次BrdU.于术后28d灌注取脑、冷冻切片,行BrdU/NF-200免疫荧光双标检测.结果 海马齿状回内BrdU阳性细胞数,给药组和对照组切割侧明显多于正常侧,给药组切割侧和正常侧分别多于对照组切割侧和正常侧;海马齿状回内的BrdU/NF-200双标神经元数,给药组切割侧最多,给药组正常侧和对照组切割侧较少,而对照组正常侧则无.结论 切割穹窿海马伞后,可致大鼠海马齿状回自体神经干细胞增殖加快并向神经元分化,脑室施加 NGF可促进其增殖和分化.