5—bromo—2—deoxyuridine标记法

以５－ｂｒｏｍｏ－２－ｄｅｘｏｙｕｒｉｄｉｎｅ标记法检测了ＳＰ２／０细胞的增殖动力学，并与３Ｈ－ＴｄＲ掺入法作了比较，结果表明本法是可靠的，且有很高的特异性。该法较３Ｈ－ＴｄＲ掺入的传统方法简便、敏感、精确、可广泛应用于细胞增殖动力学、肿瘤生物学、遗传学、分子生物学等多个领域。     

快速AS—PCR技术用于血小板HPA—1,—2,—3,—4,—5？…

目的 研究建立快速等位基因特异性（ＡＳ）引物－ＰＣＲ技术,同时对人血小板同种异型抗原系统（ＨＰＡ－１,２,３,４,５０等位基因进行分型。方法 设计合成１５闰基因特异性引物,摸索最适引物浓度、Ｍｇ＾＋＋浓度及护增参数。用该技术对１００名北京地区健康献血者ＨＰＡ－１￣５系统等位基因进行分型。结果 从１００名北京地区献血员观察到的ＨＰＡ基因频率分别是ＨＰＡ－１ａ和１ｂ为０．９９５和０．００５,ＨＰＡ－２     

下颌—面—眼—颅骨发育不良综合征一例

病例报告 史×,男,7岁。因身体矮小于1984年11月26日来诊。第5胎足月顺产,出生时有2个小而软的下切齿,3岁后换出两个新的牙齿,余乳齿未脱换。6月会笑,8月会坐,3岁会走,4岁会说简单话,7岁可数10个数。母妊娠3月时患黄疸性肝炎,服中药2月后愈。父母非近亲结婚,家族中无类似病人。 体检:发育营养差,身高84.5cm,坐高61cm,上部量49cm,下部量35.5cm,指距82.5cm,体重11公斤,颅围45.5cm。     

NK—CTL——一种新型的HLA—E限制性T细胞亚群

NK-CTL是表达HLA—I类分子特异性抑制性受体的CD8 CTL的亚群之一。NK-CTL通过其TCR，识别HLA——一种非经典的HLA—I类分子，启动机体的免疫应答，在抗肿瘤免疫、抗感染免疫以及移植排斥反应中发挥一定作用。     

Smith—Lemli—Opitz综合征研究进展

近年发现，一种以先天性多发畸形为特征的Smith—Lemli—Opits综合征在胆固醇生物合成存在缺陷，该病做为遗传代谢病的一个新分支的代表病种引起多学科研究人员的兴趣，在诊断手段致病机制、产前诊断、动物模型方面都取得了长足的进展，本文对此做一综述。     

LNT—rIL—2免疫原性研究

以家兔为实验动物，经同剂量ＬＮＴ－ｒＩＬ－２和ｒＩＬ－２多次免疫，ＥＬＩＳＡ法分析两者免疫血清中抗体滴度，结果显示：ＬＮＴ－ｒＩＬ－２免疫血清中特异抗体滴度与ｒＩＬ－２对照组无显著性差异，但总体水平有一定程度下降；ＬＮＴ呈现弱免疫原性。     

SIL—2—R与胃癌

可溶性白细胞介素２受体（ＳＩＬ－２Ｒ）是由细胞膜表面的ＩＬ－２Ｒ脱落入血而形成，在许多血液系统肿瘤、结直肠癌及肺癌时可出现增高，胃癌病人ＳＩＬ－２Ｒ增高，ＳＩＬ－２Ｒ可与ｍＩＬ－２Ｒ竞争与ＩＬ－２的结合，中和活化Ｔ细胞周围的ＩＬ－２，影响ＩＬ－２依赖的免疫应答。ＳＩＬ－２Ｒ可作为胃癌诊断中一种间接的肿瘤标记物。ＳＩＬ－２Ｒ在胃癌过继免疫治疗中有重要作用。     

IFN—γ调节HHV—6诱导单核细胞产生IL—10和IL—12

为探讨人疱疹病毒６型（ＨＨＶ－６）感染对单核／巨噬细胞功能的影响，用逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）和双抗体夹心ＥＬＩＳＡ法检测ＩＬ－１０及ＩＬ－１２的产生。ＨＨＶ－６诱导单核细胞、ＴＨＰ－１细胞表达及产生ＩＬ－１０和ＩＬ－１２。细胞因子ｍＲＮＡ动力学观察显示ＩＬ－１２随ＩＬ－１０ｍＲＮＡ积累而减少，用抗人ＩＬ－１０的单克隆抗体阻断ＨＨＶ－６诱导的内源性ＩＬ－１０，ＩＬ－１２ｍＲＮＡ表达和因子产生明显增加，提示内源性ＩＬ－１０抑制ＩＬ－１２产生。在ＨＨＶ－６感染培养中加入ＩＦＮ－γ，ＩＬ－１２基因表达随ＩＦＮ－γ浓度而增加，ＩＬ－１０ｍＲＮＡ测减少，ＩＬ－１２和ＩＬ－１０因子测定结果亦明显增加或降低，表明ＩＦＮ－γ在转录水平调节ＨＨＶ－６诱导的ＩＬ－１２和ＩＬ－１０产生，其机理可能主要与ＩＦＮ－γ抑制内源性ＩＬ－１０有关。     

大鼠脑中催产素和加压素免疫反应性物质的分布

用ABC法研究大鼠脑中催产素(OT)和加压素(VP)免疫反应性物质的分布。阳性细胞主要局限于下丘脑,而阳性纤维则广泛分布于下丘脑和脑干等部位。OT和VP的分布情况有差别。在视上核和室旁核大细胞后部,VP阳性大细胞明显地比OT大细胞密集;同样,在向正中隆起方向行走的阳性纤维束中,VP纤维也占大多数。而在室旁核尾部的小细胞内侧部,中小型OT阳性细胞比VP细胞密集;并且,在脑干的孤束核、三叉神经脊束核、迷走神经背核、疑核、臂旁核和蓝斑核中,OT阳性纤维也占主要地位。实验结果提示,下丘脑的OT和VP能神经元除参与垂体后叶内分泌外,还可能直接支配一些脑干内脏核。推测在正常生理状态下,垂体后叶的VP分泌可能比OT分泌占有更重要的地位;而OT则可能在充当神经递质或调制物在参与下丘脑对脑干内脏核团的调节中有更重要的位置。     

Vesicular glutamate transporters type 1 and 2 expression in axon terminals of the rat nucleus of the solitary tract

The nucleus of the solitary tract is the site of termination of primary afferent fibers running in the facial, glossopharyngeal and vagus nerves. The present study was performed to map the distribution of glutamatergic axons terminals in the rat nucleus of the solitary tract using immunodetection of vesicular glutamate transporter 1 and vesicular glutamate transporter 2. The two vesicular glutamate transporters were differentially distributed among nucleus of the solitary tract subdivisions. Vesicular glutamate transporter 1 immunoreactivity was mostly found in the lateral part of the nucleus (ventrolateral, interstitial and intermediate subdivisions) whereas vesicular glutamate transporter 2 labeling was distributed throughout the nucleus of the solitary tract. Electron microscope examination indicated that vesicular glutamate transporter immunoreactivity was localized in axon terminals filled with round synaptic vesicles. After injection of cholera toxin B subunit in sensory ganglia, anterograde labeling was found in vesicular glutamate transporter 1, as well as vesicular glutamate transporter 2-immunoreactive boutons. Double immunolabeling experiments allowed distinctions between terminals expressing either vesicular glutamate transporter 1 or vesicular glutamate transporter 2 or both vesicular glutamate transporter 1 and vesicular glutamate transporter 2 immunoreactivities. The latter population, expressing both transporters immunolabeling, completely disappeared after deafferentation induced by removal of sensory ganglia. This study indicates that vesicular glutamate transporter content identifies three different subpopulations of glutamatergic boutons in the nucleus of the solitary tract and provides definitive evidence that primary afferent neurons contribute glutamatergic terminals to the nucleus of the solitary tract.     

鸡迷走神经传入纤维在中枢的投射

选用12只1～2kg重的来杭鸡,于颈部迷走神经干内注射CB-HRP,TMB法成色。结果见被标记的传入纤维主要分布于孤束和孤束核;在迷背核、孤束核和第四脑室膜三者之间分布有大量的标记终末和迷背核神经元树突;对侧孤束和孤束核中的标记范围与注射侧的相似,但较疏淡。此外,在三叉神经脊束、楔外侧核及第1、2颈髓的背索和背索核中发现有明显的标记终末;在迷背核中也有少量传入标记。     

内脏伤害性传入信息向三叉神经脊束间质核的投射

舌咽、迷走神经向三叉神经脊束间质核的投射纤维除传递非伤害性内脏信息者外是否也含内脏伤害性传入？尚未见报道。本文综合应用：（1）福尔马林刺激舌咽、迷走神经支配的一些内脏诱导FOS蛋白表达；（2）抽除外周神经节观察三叉神经脊束间质核内一些与伤害性信息传递有关的神经递质的变化；（3）运用荧光金逆行追踪与免疫组化相结合的双重反应等方法，对大鼠投射至三叉神经脊束间质核的舌烟、迷走神经初级传入纤维中是否含传递伤害性刺激信息的成分进行了研究。结果证明：(1)2％福尔马林刺激大鼠软腭、咽、食道和胃后，三叉神经脊束间质核内有大量FOS阳性神经元表达，这些阳性神经元与舌咽、迷走神经初级传入分布区基本一致。（2）抽除一侧舌咽、谈走神经报及其神经节后，术侧三叉神经脊束间质核内SP、CGRP等和痛传递有关的神经肽及BSI-B_4阳性终末与对侧比较明显减少。（3）荧光金微量注入三叉神经脊束间质核后，在舌咽、迷走神经节的下节内发现被荧光金逆标的细胞中分别有10％和12％呈SP、CGRP阳性，此两种双重阳性细胞分别占SP、CGRP阳性细胞的33．3%和278％。以上结果提示，三叉神经脊束间质核接受舌咽、迷走神经的传入纤维中有一部分传递内脏伤害性信息。     

Properties of solitary tract neurons receiving inputs from the sub-diaphragmatic vagus nerve

Vagal afferents ascending from the gastrointestinal tract synapse on neurons in the nucleus of the solitary tract. Although these neurons constitute a significant proportion of solitary tract cells their firing behaviour and synaptic properties are not documented. Since gastrointestinal tract afferent termination sites overlap with regions mediating cardiorespiratory reflexes the possibility of convergence with afferents mediating cardiovascular and respiratory reflexes was proposed. Here we describe some electrophysiological and morphological properties of solitary tract neurons orthodromically driven from the subdiaphragmatic vagus nerves and assess possible convergent inputs from cardiorespiratory afferents. Whole-cell recordings of solitary tract neurons responding to electrical stimulation of the sub-diaphragmatic vagus nerves (0.1-1 ms; 1-10 V; 2-20 Hz) were made in a working heart-brainstem preparation of rat. Baroreceptors were stimulated by raising pressure in the aorta or carotid sinus, whereas aortic injection of sodium cyanide (0.05% solution 25-50 microl) was used to activate peripheral chemoreceptors. Phrenic nerve activity and heart rate were monitored continuously. Of 88 solitary tract neurons tested, 39 responded with an evoked excitatory synaptic potential following stimulation of the sub-diaphragmatic vagus nerves. Resting membrane potential and input resistance of sub-diaphragmatic vagus nerve driven solitary tract neurons were 53.2 +/- 0.5 mV and 291 +/- 17 Mohms, respectively (mean +/- S.E.M.). Response latencies to sub-diaphragmatic vagus nerve stimulation were divided into two groups: <20 ms (16.0 +/- 2 ms, n = 7; mean +/- S.E.M.) and >20 ms (77.3 +/- 5 ms, n = 32). One additional neuron displayed an evoked inhibitory postsynaptic potential (latency 175 ms). Nineteen neurons showed ongoing activity which consisted of either irregular single action potential firing (0.5-10 Hz; n = 12) or burst discharge (n = 7). Of 33 neurons tested, 17 showed spike frequency adaptation during injection of positive current, whereas 19 of 38 cells displayed rebound excitation following release from hyperpolarized potentials. There was no correlation between these properties and synaptic latencies. Ninety-one per cent of neurons tested displayed synaptic depression following paired pulse stimulation of the sub-diaphragmatic vagus nerve over intervals up to 500 ms. Stimulation of either baroreceptors (n = 31) or chemoreceptors (n = 36) failed to elicit a synaptic response in all sub-diaphragmatic vagus nerve-driven solitary tract neurons. Neurobiotin-labelled solitary tract neurons (n = 10) were from both latency groups and were located medial to the solitary tract at the level of area postrema, -0.3 mm to +1 mm from the obex. One cell was located in commissural subnucleus at midline, seven cells dorsal to the tractus solitarius and three ventral and medial to it. Soma sizes were 23 +/- 9.6 x 14 +/- 4.9 microm (range: 50 x 16 microm to 15 x 7 microm). The number of primary dendrites varied from three to five, secondary from one to eight and tertiary zero to four. Labelled axons were found in seven cells which ramified extensively in the solitary tract nucleus (n = 3) and/or branched extensively in the dorsal vagal motonucleus (n = 3) and/or projected towards the ventrolateral medulla (n = 3). We conclude that solitary tract neurons receiving signals from the sub-diaphragmatic vagus nerves (most likely from gastrointestinal tract structures) appear to be a distinct pool of neurons. There was a heterogeneity in terms of both their ongoing activity and projection targets but despite this, there were three consistent properties. First, sub-diaphragmatic vagus nerve evoked predominantly excitatory synaptic responses in solitary tract neurons; second, neurons exhibited lasting paired pulse depression following activation of sub-diaphragmatic vagus nerves; and third, sub-diaphragmatic vagus nerve-driven solitary tract neurons were     

亮氨酸脑啡肽样和P物质样免疫反应物在大鼠延髓迷走神经背核簇和网状结构内的分布

用PAP法研究亮氨酸脑啡肽样(L—ENK—LI)和P物质样(SP—LI)免疫反应物在大鼠延髓迷走神经背核簇和网状结构内的分布。证明在孤束核、迷走神经背核、外侧网状核外侧部及其外侧的区域内有大量的L-ENK样终末和纤维,在背侧和腹侧网状核之间的移行区内有中等量的分布,其余区内为少量。L-ENK样胞体在孤束核、咀侧腹外侧网状核、巨细胞网状核的腹侧部和α部、外侧旁巨细胞核以及中缝大核内均有许多分布,在迷走神经背核的尾侧部、背侧和腹侧网状核之间的移行区有中等量,其余区内为少量。SP样反应物的分布与L-ENK样物类似,但其终末和纤维的数量较L-ENK者略少,阳性胞体的数量除了在中缝核及外侧旁巨细胞核内侧端中的量较L-ENK样胞体多以外,在其余区内均较少。     

神经元核抗原在大鼠中枢神经系统神经元的不同表达

目的观察正常大鼠中枢神经系统(CNS)内神经元核抗原(NeuN)的表达。方法正常成年SD大鼠5只,常规固定,恒冷箱切片,免疫组织化学、免疫荧光染色、尼氏染色。结果NeuN在成年大鼠CNS神经元有4种表达:大部分神经元(大脑皮质、基底核、脊髓背角等)有明显表达;部分神经元为中等表达(三叉神经脊束核、下丘脑室旁核等);部分神经元为弱表达(孤束核、蓝斑等);而有些核团神经元缺乏NeuN的表达(视上核、弓状核、迷走神经背核等)。结论中枢神经系统不同区域或核团的神经元NeuN的表达程度不同,有的区域或核团神经元缺乏NeuN表达。     

大鼠胸腺传入纤维在中枢内的跨节投射——CB-HRP法

本文将CB-HRP注入大鼠胸腺,研究了标记细胞在感觉神经节的分布及初级感觉终末在脊髓、脑干中的投射。结果表明:1.在双侧迷走神经结状节中有标记细胞;2.在C_1-T_5段双侧脊神经节中有标记细胞,推测这些标记细胞可能随交感神经传入,其中C_3-C_4段的投射可能随膈神经传入;3.脊髓中的标记终末,见于C_1-C_8节段,主要分布在Ⅰ层内;4.脑干中的标记终末,主要见于双侧孤束核的全长及连合核中。在疑核标记的节前神经元附近有的也可见终末分布,提示胸腺的一级传入与传出神经元间可能存在直接联系。     

猫降结肠传出和传入神经元的节段性分布—HRP法研究

本文用HRP法研究了猫降结肠的传出和传入神经元节段性分布,结果表明:1.猫降结肠的副交感中枢位于双侧骶髓1—3节段,左侧占优势。标记细胞主要位于网状核,其次为中间带外侧核,细胞均为中、小型。2.在双侧腰髓1—4节段的中间带外侧核见到许多标记细胞。3.降结肠感觉神经纤维来自双侧T_(13)—L_5和L_7—Co_1脊神经节,其细胞形态多为圆形、椭圆形,以中,小型细胞为主。4.感觉神经元的中枢突经Lissauer束进入骶髓1—3节段,围绕后角形成较粗大的外侧束和较细的内侧束。外侧束到达脊髓灰质Ⅴ、Ⅶ层和灰质后连合,内侧束到达后角灰质第Ⅱ、Ⅲ层。