应用多电极记录技术研究海马切片癫痫样放电的起始点和传播方向

弄清癫痫样放电的起始位置和传播方向对研究癫痫机制及其临床治疗有重要意义．为了解决这一问题,应用微电极阵列对低镁人工脑脊液诱导的Sprague-Dawley (SD)大鼠海马切片的癫痫样放电进行记录．分别用癫痫样放电的两种成分：场电位和多单元信号来确定癫痫样放电的起始位置和传播方向．首先计算并比较了海马切片锥体细胞层位置电极记录的癫痫样放电场电位的起始时间,由起始时间的先后关系确定癫痫样放电在锥体细胞层的起始位置和传播方向．然后用整个切片上记录的癫痫样放电的多单元信号动作电位序列进行互相关分析,进一步确定了癫痫样放电在整个海马切片内的起始位置和传播方向．结果显示,CA3区的癫痫样放电具有比CA1区更高的幅度和更长的持续时间,表明CA3区有更高的兴奋性．对于记录到的同步癫痫样放电,CA3b区场电位和多单元信号均比CA3c和CA1区出现更早,起始位置和其随后位置之间的传播延　时与二者之间的距离成正相关．因此,在低镁模型的大鼠海马切片中,癫痫样放电起始于CA3b区并分别向CA3c和CA1区传播．     

马桑内酯在Wistar大鼠海马脑片上引起的痫样放电及SC_(1001)钠盐的对

马桑内酯(CoriariaLactone,CL)是从抗精神病中草药—马桑寄生中提取的有效成分,它具有致痫作用。目前,运用CL在整体动物身上建立急、慢性癫痫模型的实验已获成功。为了进一步从细胞水平探讨CL的致痫作用及可能的电生理机制,并观察抗癫新药SC1001钠盐对CL所致痫样放电活动的抑制作用,本实验采用离体海马脑片技术,在59例Wistar大鼠的海马脑片标本上进行了以下研究:以海马CA1区锥体细胞诱发场电位为指标,观察了不同刺激强度时顺向、逆向刺激与诱发场电位之间的关系;观察了持续灌流人工脑脊液(ACSF)对诱发场电位的影响和双脉冲易化,频率…     

钩藤对致痫大鼠海马脑片诱发场电位的影响

目的研究钩藤对癫痫模型海马脑片诱发场电位的影响。方法以毛果芸香碱致痫大鼠为实验对象,采用脑片旁滴注给药,用细胞外玻璃微电极记录方法,观察钩藤对癫痫模型离体海马脑片CA1区锥体细胞诱发群锋电位（populationspike,PS）的影响。结果给予钩藤后使致痫大鼠海马脑片PS幅度平均降低27.64％,平均8.71min恢复(n=14,P<0.01)。结论钩藤能降低致痫大鼠海马脑片CA1区顺向诱发PS幅度,提示钩藤对中枢神经系统的突触传递过程有明显的抑制效应,具有抗癫痫作用。     

海马脑片抗癫痫药物研究的离体模型

目的：建立离体海马脑片癫痫样放电模型并用于抗癫痫药物研究。方法：在豚鼠海马脑片上灌流青霉素建立颠阗痫样放电的离体模型。并用此模型对抗癫痫药物苯巴比妥钠和苯妥英钠两种药物在不同浓度下对癫痫样放电的对抗作用进行了定量分析,结果：在海马脑片上灌流致痫药物可建立一个较好的离体组织癫痫样放电模型,苯的对抗作用进行了定量分析,结果：在海马脑片上灌流致痫药物可建立一个较好的离体组织癫痫样放电模型。苯巴比妥和苯妥英钠在一定浓度下均有显著对抗癫痫样放电的作用,且与整体实验的结果相一致。结论：本实验建立有离体脑片模型具有实验手段简单,方法灵活,易于建立药物量效关系等优点,可用于抗癫痫药物筛选和研究。     

羟基马桑毒素所致的大鼠海马锥体细胞痫样放电活动

本研究采用离体海马脑片电生理研究技术,细胞外记录海马锥体细胞群体锋电位(population spike,PS),观察羟基马桑毒素(tutin)对大鼠海马脑片CA1区锥体细胞电活动的影响,探讨tutin是否具有致痛作用及其致痫机制。结果如下:(1)用40、30和20μg/ml浓度的tutin灌流海马脑片,可显著增高由顺向刺激Schaffer侧支所诱发的PS的幅度,灌流tutin 30min时,PS第一个波的幅度分别为对照的(388.7±20.1)%、(317.2±19.1)%和(180.9±11.6)%(各组n=5,P<0.05)。(2)伴随PS波幅的增高,可出现成串痫样放电波,波数4～11个不等。(3)灌流tutin后的部分脑片(n=9/34),在未刺激Schaffer侧支时也出现自发的成串、高幅痫样放电。(4)灌流CNQX阻断非NMDA受体后,再灌流tutin,PS幅度和放电波数均无显著性变化,即CNQX可完全抑制tutin所致的痫样放电;灌流AP-5阻断NMDA受体后,tutin仍可使PS幅度增高但放电波数无显著性增加,即AP-5可部分抑制tutin所致的痫样放电。上述结果表明,tutin可使海马脑片锥体细胞兴奋活动增强,具有致痫作用;兴奋性谷氨酸受体尤其是非NMDA受体可能介导tutin的致痫作用。     

大鼠多通道在体记录

多通道在体记录技术,能在自由活动的动物脑内,观察和记录局部脑区群体神经元的活动状况,是分析大脑神经信息编码的有力工具。要开展多通道在体记录研究,多电极阵列驱动器的设计非常关键,也是实现该技术的一大难点。根据转动螺杆推动螺帽移动的机械驱动原理,作者设计了适合大鼠多通道在体记录的、独立可调式16道电极阵列驱动装置。通过该装置,可对16道记录电极中的任意一道进行独立驱动,从而控制每根记录电极在大鼠大脑中的垂直记录位置。运用该多电极阵列驱动装置,对大鼠单侧海马脑区的多通道在体记录表明,在大鼠海马CA1区,存在不同放电波形和放电模式的神经元,它们分别与海马CA1区的锥体神经元和中间神经元相对应。一般锥体神经元动作电位的放电波形较宽,放电频率则较低。在海马CA1区还存在编码空间环境中特定位置信息的神经元,被称为位置细胞。这些位置细胞在某一空间环境中有各自对应的反应区域,在该区域内位置细胞的放电频率增加,在区域外则基本维持在一较低的活动水平。     

胍丁胺对大鼠海马 CA1区神经元放电的影响

应用细胞外记录单位放电技术,在大鼠海马脑片上观察了胍丁胺(agmatine,Agm)对CAl区神经元放电的影响。实验结果如下：(1)在47个海马脑片放电单位上灌流Agm(0．1—1．0μmol／L)2min,有38个单位(80．9％)自发放电频率明显降低,且呈剂量依赖性,9个单位(19．1％)无明显的反应;(2)预先用0．2mmol／L的L-谷氨酸(L-glutamate,L-Glu)灌流12个海马脑片放电单位,有9个单位(75％)放电频率明显增加,表现为癫痫样放电,在此基础上灌流Agm(1．0μmol／L)2min,其癫痫样放电被抑制;(3)在7个海马脑片放电单位上给予L型钙通道激动剂Bay K8644(0．1μmoL／L)时,有6个单位(85．7％)放电频率明显增加,另外1个单位(14．3％)无明显变化,再给予Agm(1．0μmol／L)2min,其放电频率被明显抑制;(4)13个CAl放电单位,灌流50μmoL／L一氧化氮合酶(NOS)抑制剂N^G-nitro-L-arginine methyl ester。(L-NAME)5min后其放电频率明显增加,在此基础上再给予Agm(1．0μmol／L)2min,有11个单位(84．6％)的放电频率被抑制,有2个单位(15．4％)的变化不明显。上述结果提示：胍丁胺能抑制海马CAl区神经元自发放电以及由谷氨酸、BayK8644和L-NAME诱发的放电,这一抑制效应可能与胍丁胺阻断CAl区锥体细胞上的NMDA受体,并减少钙离子内流有关。     

高频电刺激对大鼠海马脑片癫痫样放电特性影响的研究

本文采用电极阵列检测技术,在大鼠海马脑切片上诱导出稳定的癫痫样放电,分析、研究130 Hz的高频电刺激(high-frequency stimulation,HFS) CA3区时,海马切片在癫痫发作间期放电(inter-ictal discharges,IID)和发作期放电(ictal discharges,ID)的各项参数、癫痫样放电地起始位点、传播方向和传输速率以及各频段的功率谱密度.结果显示:高频电刺激可以有效地降低癫痫发作期的幅值、减少持续时间、增长潜伏时间、抑制癫痫样放电由IID向ID的转变等.提示高频电刺激抑制癫痫的作用机制是通过促进神经元之间的抑制性传输系统,并且抑制海马神经元之间的兴奋性连接,从而达到抑制效果.     

大鼠离体脑片癫痫放电特征及EC—海马环路的作用

目的和方法：采用４００～５００μｍ大鼠水平脑切片强直电刺激海马Ｓｃｈａｅｆｅｒ侧枝（６０Ｈｚ、２ｓ）全细胞、细胞外同步记录ＣＡ１神经元胞体电活动和相应树突区场电位,探讨其在癫痫发生中的作用。结果：①５３片脑片上记录到细胞内、外同步发生的原发性后放,持续２０ｓ以上,放电形式和持续时间常在第６个刺激串后趋于稳定。ＣＡ１神经元的原发性后放常跟在强直电刺激引起的阵发性去极化或超极化偏移之后（ＰＤＳ、ＰＨＳ）。它可以从紧张性放电向爆发性放电转化,振幅逐渐递增并与细胞外癫痫样放电同步,产生癫痫放电极性偏移;②其中８／４０脑片细胞外可记录到继发性后放之后出现的自发性发作样癫痫放电,长达数分钟,与全细胞记录的ＥＰＳＰ同步。切断ＥＣ与海马之间的联系可以易化海马癫痫电活动（３／５）。结论：ＥＣ输入到海马的神经通路可能在封闭的ＥＣ海马环路中起着重要的门控作用     

白藜芦醇抑制大鼠海马 CA1区神经元放电

应用细胞外记录单位放电技术,在大鼠海马脑片上观察了白藜芦醇(resveratrol)对海马CAI区神经元放电的影响。实验结果如下：(1)在52个CAI区神经元放电单位给予白藜芦醇(0．05、0．5、5μmol／L)2min,有46个放电单位(88.5％)放电频率明显降低,且呈剂量依赖性;(2)预先用0．2mmol／L的L-glutamate灌流海码腑片,8个放电单位放电频率明显增加,表现为癫痫样放电,在此基础上灌流白藜芦醇(5μmol／L)2min,其癫痫样放电被抑制;(3)预先用L型钙通道开放剂Bay K8644灌流7个海马5脑片,有6个单位(85.7％)放电增加,在此基础上灌流白藜芦醇(5μmol／L)2min,其放电被抑制;(4)9个放电单位灌流一氧化氮合酶抑制剂L-NAME(N^0-nitro-L-arginine methylester)50μmol／L,有7个单位(77．8％)放电明显增加,在此基础上灌流白藜芦醇(5μmol／L)2min,放电被抑制;(5)10个放电单位灌流人电导钙激活性钾通道阻断剂TEA(tetraethylarnmonium chloride)1mmol/L后,有9个单位(90％)放电增加,在此基础上灌流白藜芦醇(5μmol／L)2min,8个放电单位(88,9％)放电频率明显减低。以上结果提示：白藜芦醇能抑制海马神经元自发放电以及由L-glutamate、L-NAME、Bay K8644和TEA诱发的放电,可能与白藜芦醇抑制L型钙通道,减少钙内流有关;似乎与大电导钙激活性钾通道无关。     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Comparative morphology of the carpel in the Liliaceae: Hewardieae, Petrosavieae, and Tricyrteae

The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle.     

Entwicklungsgeschichte und Ultrastruktur von Pollenkitt und Exine bei nahe verwandten entomophilen und anemophilen Angiospermensippen derAlismataceae,Liliaceae, Juncaceae,Cyperaceae, Poaceae undAraceae

Some closely related members of the monocotyledonous familiesAlismataceae, Liliaceae, Juncaceae, Cyperaceae, Poaceae andAraceae with variable modes of pollination (insect- and wind-pollination) were studied in relation to the ultrastructure of pollenkitt and exine (amount, consistency and distribution of pollenkitt on the surface of pollen grains). The character syndromes of pollen cementing in entomophilous, anemophilous and intermediate (ambophilous or amphiphilous) monocotyledons are the same in principal as in dicotyledons. Comparing present with former results one can summarize: 1) The pollenkitt is always produced in the same manner by the anther tapetum in all angiosperm sub-classes. 2) The variable stickiness of entomophilous and anemophilous pollen always depends on the particular distribution and consistency of the pollenkitt, but not its amount on the pollen surface. 3) The mostly dry and powdery pollen of anemophilous plants always contains a variable amount of inactive pollenkitt in its exine cavities. 4) A step-by step change of the pollen cementing syndrome can be observed from entomophily towards anemophily. 5) From the omnipresence of pollenkitt in all wind-pollinated angiosperms studied one can conclude that the ancestors of anemophilous angiosperms probably have been zoophilous (i.e. entomophilous) throughout.

     

Identification and Characterization of the First Equine Parainfluenza Virus 5

正Dear Editor,Parainfluenza virus 5 (PIV5), known as canine parainfluenza virus in the veterinary field, is a negative-sense,nonsegmented, single-stranded RNA virus belonging to the Paramyxoviridae family (Chen 2018). The virus was first reported in primary monkey kidney cells in 1954 (Hsiung1972), then it has been frequently discovered in various