雌激素对电刺激诱发杏仁核多巴胺释放的影响

用快速周期伏安法（FCV）测定了去卵巢（OVX）雌鼠,经雌激素处理的去卵巢雌鼠及正常雌鼠的中央杏仁核（CAN）的多巴胺（DA）释放,并应用放射免疫技术检测了大鼠在不同处理条件下血清雌二醇（E2）含量,结果表明,雌激素处理的OVX组大鼠的杏仁核DA释放量均高于对照OVX组大鼠,并随苯甲酸雌二醇（EB）注射剂量的增加,DN释放量及血清E2含量迹增加,有明显的量－效关系,提示雌激素可能是调节大鼠CAN区DA释放量的重要因素。     

脑内多巴胺信号动态变化的检测方法及在成瘾研究中的应用

多巴胺是脑内重要的神经递质,中脑多巴胺系统在控制奖赏、动机、运动和情绪调节过程中起重要作用。脑内多巴胺功能紊乱与药物依赖、精神分裂症、抑郁症和帕金森氏病等神经精神障碍有关,动态测定脑内多巴胺变化对于了解多巴胺功能和揭示相关疾病病理机制具有重要意义。本文主要介绍了微透析、快速扫描循环伏安法和光纤光度法的基本原理和方法,并对比分析了这些技术在多巴胺动态检测应用中的优缺点。以药物成瘾研究为应用实例,利用微透析法发现伏隔核壳部是成瘾性药物产生奖赏效应的关键部位,快速扫描循环伏安法检测到与可卡因自身给药行为相关的三种多巴胺信号模式,而光纤光度法则揭示了酒精成瘾和复吸过程中伏隔核和中脑复侧被盖区多巴胺活动特征存在空间和时间上的多样性。这些发现为揭示药物成瘾的机制做出了重要贡献。     

18O标记法在定量蛋白质组学中的应用

基于质谱技术去识别和检测蛋白质表达差异是一个热点,有助于生物过程和体系的分子机制的研究。近年来各种基于质谱技术的定量蛋白质组学研究方法发展较快,相对其他方法而言,18O标记法是一种较为理想、相对容易实现并且在不断完善的体外标记方法,最近在定量蛋白质组学研究中应用较多。现对18O标记法原理、特点以及技术方法的优化和应用进展进行综述。     

实时荧光定量PCR技术在鱼类病害研究中的应用

实时荧光定量PCR技术是一种新的核酸定量技术,通过检测PCR产物中荧光信号强度达到定量的目的,与常规PCR相比,具有无污染、特异性强、检测灵敏、定量准确等特点,该技术在分子诊断、动植物检疫等方面得到了广泛的应用.目前水产养殖业处于飞速发展时期,其中鱼类的病害问题也日益突出,为了预防和控制鱼类病害,实时荧光定量PCR技术已逐渐应用于鱼类病害的研究中.该文将从实时荧光定量PCR的技术原理、主要类型以及实时荧光定量PCR技术在鱼类病害研究的应用研究作一综述.     

番茄叶片基因组DNA快速制备技术及其在基于实时荧光定量PCR的转基因检测中的应用

以番茄(Solanum lycopersicum L.cv.Micro-Tom)叶片为试材,建立了一种简便快速制备叶片基因组DNA的方法。2~20 mm2的叶片即可满足制备要求,制备过程只需一种提取试剂、只涉及1次移液和1次离心操作,不涉及沉淀。确定了所制备的DNA用于实时荧光定量PCR的合适用量为0.1~0.2μL(反应总体积为12.5μL),发现过量模板的使用可降低PCR效率且可导致扩增失败。该项DNA快速制备及相适应的实时荧光定量PCR技术已成功应用于番茄转基因植株检测。     

一种快速、灵敏的血小板释放功能检测方法及其在抗血小板药物研究中的应用

本文报道了一种快速、灵敏的血小板释放功能检测方法:利用荧光素-荧光素酶在有ATP、Mg~(2+)、O_2存在时产生的生物发光素测定血小板ATP的释放量,以反映血小板的释放功能;研究了ADP、AA、胶原、凝血酶等四种诱导剂对血小板释放功能的作用,发现ADP的诱导释放能力较其他三者为弱;观察在不同剂量ADP和AA的诱导下,血小板聚集强度和释放能力之间的关系,研究了血小板数等因素对ATP释放功能测定的影响。应用该方法研究了Aspirin及活血化淤药物川芎嗪,毛冬青甲素对血小板释放功能的影响,发现Aspirin对AA诱导的释放反应有强烈的抑制作用。在以ADP诱导的释放反应中,川芎嗪的抑制作用较毛冬青甲素更为强烈。     

检测脊髓灰质炎IgM抗体捕捉法ELISA的建立及其在早期快速诊断中的应用

首次建立了用于脊髓灰质炎快速、分型诊断的血清IgM抗体捕捉法ELISA。检测了52例疑似病人,IgM阳性率为76.9％(40/52),而粪便病毒分离率仅为44.2％(23/52),前者的阳性检出率明显高于后者。做病毒分型诊断结果,与分离病毒中和试验定型的总符合率为92.86％(39/42)。检测601例来自全国各省的脊灰疑似病人血清,其阳性率波动于13％～92.3％,平均为61.1％。阳性率与收集血清的病日相关,发病0～3天收集者,阳性检出率为69.5％,4～25天者为64％,26～54天者为52％,55天以上者则未能检出。本检测方法需时1.5天,简便、敏感、特异,重复性良好,适用于脊灰的早期快速诊断。对其实用性进行了讨论。     

洗胃法在鳄蜥食性研究中的应用

为了研究洗胃法是否适用于鳄蜥（Shinisaurus crocodilurus）的食性研究,2006年8～9月我们在广东罗坑省级自然保护区,通过投喂食物,然后再冲洗的方法来检验洗胃法的冲洗效果。结果显示洗胃法对成体和亚成体鳄蜥的总洗出率均高于99．30％,食物个体数的洗出率均高于90．83％,投喂的12种食物都能被洗出。观察发现洗胃之后,对鳄蜥的正常生活没有影响。鳄蜥吃进的实验食物中,未洗出的食物大小与鳄蜥的体长、头宽、体重不相关;食物的洗出效率与鳄蜥的体长、头宽、体重不相关。成体、亚成体洗出和未洗出的食物大小均存在显著差异。未被洗出的食物个体在12种食物中不是随机分布的,体积大的食物比体积小的食物容易被洗出。研究表明,洗胃法适用于鳄蜥的食性研究,并且效果较好。     

改良胃管置入法在昏迷气管切开病人护理中的应用研究

目的：探讨改良后胃管置入法在昏迷气管切开病人护理中的应用。方法：患者采取平卧位,保持头部、颈项、躯干在同一水平线上。置入胃管之前,先清理口、鼻、气管内分泌物。根据患者情况,选择合适型号硅胶胃管。操作者量好胃管置入长度后,用液体石蜡充分润滑胃管,左手托住胃管后端,右手持镊子夹住胃管前端,沿一侧鼻孔鼻中隔缓慢插入到合适距离,固定胃管。胃管末端连接注射器抽吸,如有胃液抽出,表示已插到胃内。结果：所有病例,均采用此种方法。24例病例中,一次性操作成功18例（占75．0％）,4例（占16．7％）病例应用此法两次后成功置入胃管,2例（占8.3％）患者应用传统昏迷病人胃插管术三次均未获成功,改用此法后一次即成功。结论：改良胃管置入法操作简单、实用、安全,在昏迷气管切开病人护理中是促进患者早日康复的重要措施.     

改良胃管置入法在昏迷气管切开病人护理中的应用研究

目的:探讨改良后胃管置入法在昏迷气管切开病人护理中的应用。方法:患者采取平卧位,保持头部、颈项、躯干在同一水平线上。置入胃管之前,先清理口、鼻、气管内分泌物。根据患者情况,选择合适型号硅胶胃管。操作者量好胃管置入长度后,用液体石蜡充分润滑胃管,左手托住胃管后端,右手持镊子夹住胃管前端,沿一侧鼻孔鼻中隔缓慢插入到合适距离,固定胃管。胃管末端连接注射器抽吸,如有胃液抽出,表示已插到胃内。结果:所有病例,均采用此种方法。24例病例中,一次性操作成功18例(占75.0%),4例(占16.7%)病例应用此法两次后成功置入胃管,2例(占8.3%)患者应用传统昏迷病人胃插管术三次均未获成功,改用此法后一次即成功。结论:改良胃管置入法操作简单、实用、安全,在昏迷气管切开病人护理中是促进患者早日康复的重要措施。     

In vivo electrochemical evidence for simultaneous 5-HT and histamine release in the rat substantia nigra pars reticulata following medial forebrain bundle stimulation

Exploring the mechanisms of serotonin [5-hydroxytryptamine (5-HT)] in the brain requires an in vivo method that combines fast temporal resolution with chemical selectivity. Fast-scan cyclic voltammetry is a technique with sufficient temporal and chemical resolution for probing dynamic 5-HT neurotransmission events; however, traditionally it has not been possible to probe in vivo 5-HT mechanisms. Recently, we optimized fast-scan cyclic voltammetry for measuring 5-HT release and uptake in vivo in the substantia nigra pars reticulata (SNR) with electrical stimulation of the dorsal raphe nucleus (DRN) in the rat brain. Here, we address technical challenges associated with rat DRN surgery by electrically stimulating 5-HT projections in the medial forebrain bundle (MFB), a more accessible anatomical location. MFB stimulation elicits 5-HT in the SNR; furthermore, we find simultaneous release of an additional species. We use electrochemical and pharmacological methods and describe physiological, anatomical and independent chemical analyses to identify this species as histamine. We also show pharmacologically that increasing the lifetime of extracellular histamine significantly decreases 5-HT release, most likely because of increased activation of histamine H-3 receptors that inhibit 5-HT release. Despite this, under physiological conditions, we find by kinetic comparisons of DRN and MFB stimulations that the simultaneous release of histamine does not interfere with the quantitative 5-HT concentration profile. We therefore present a novel and robust electrical stimulation of the MFB that is technically less challenging than DRN stimulation to study 5-HT and histamine release in the SNR.     

电剌激及心肌缺血所致豚鼠心脏神经肽Y释放变化的研究

电剌激豚鼠心脏左星状交感神经节,可引起钙依赖性的神经肽Ｙ（ＮＰＹ）胞外释放,缺血１０ｍｉｎ后行电刺激（Ｓ２）,与对照期（Ｓ１）比较,ＮＰＹ释放无明显变化,缺血２０ｍｉｎ后行电剌激,ＮＰＹ释放受到一定程度的抑制,（Ｓ２／Ｓ１：０．７２,Ｐ＜０．０５）,而再灌注５ｍｉｎ后行电剌激ＮＰＹ释放抑制作用逐步消失,与Ｓ１比较,已无明显减少（Ｓ２／Ｓ１;１．０１,Ｐ＞０．０５）。仅缺血无电剌激几乎不引起ＮＰＹ释放。这些结果提示,电剌激交感神经节可致ＮＰＹ呈现一种钙依赖性出胞释放,缺血早期ＮＰＹ释放受到某些代谢产物抑制,恢复再灌注后,代谢产物逐步冲洗出,抑制作用也随之消失。     

AN ADAPTATION OF THE PUSH-PULL CANNULA METHOD TO STUDY THE IN VIVO RELEASE OF [3H]DOPAMINE SYNTHESIZED FROM [3H]TYROSINE IN THE CAT CAUDATE NUCLEUS: EFFECTS OF VARIOUS PHYSICAL AND PHARMACOLOGICAL TREATMENTS

Abstract— The release of [³H]dopamine ([³H]DA) continuously synthesized from l-[3,5–³H]tyrosine from the caudate nucleus of the cat was estimated in halothane anaesthetized or‘encéphale isolé’animals. For this purpose, an improved superfusion cannula, avoiding tissue damage, was used. The best localization for the tip of the superfusion cannula was found first by determining the topographical distribution of endogenous DA within the caudate nucleus. A rostro-caudal heterogenous distribution of the transmitter was detected. In perfusion experiments, l-[3,5–³H]tyrosine was introduced continuously at a rate of 33μl/min. [³H]DA was the only catecholamine found in serial 15 min fractions as revealed by cochromatography. The spontaneous release of [³H]DA was greater in anaesthetized than in ‘encéphale isolé’ cats; it represented 150 and 100 times the blank value, respectively. Depolarization by K⁺ (30 mm) applied locally in the striatum or by electrical or mechanical stimulation of the substantia nigra caused a transitory increase in [³H]DA release. Conversely, a decrease in nerve activity induced by tetrodotoxin (5 × 10^?-⁷ m) or by electrocoagulation of the substantia nigra was associated with a decline in the amounts of [³H]DA in superfusates. A temporary reduction in [³H]DA release could also be obtained by a short-lasting cooling block of the substantia nigra. As expected, d-amphetamine (10^?-⁵ m) and benzotropine(10^?-⁷ m) added to the superfusing medium increased [³H]DA release. These pharmacological results, as well as the changes in [³H]DA release observed after various manipulations of the activity of dopaminergic neurones, confirms the validity and the high sensitivity of this approach.     

Endogenous opiates and behavior: 2010

This paper is the thirty-third consecutive installment of the annual review of research concerning the endogenous opioid system. It summarizes papers published during 2010 that studied the behavioral effects of molecular, pharmacological and genetic manipulation of opioid peptides, opioid receptors, opioid agonists and opioid antagonists. The particular topics that continue to be covered include the molecular-biochemical effects and neurochemical localization studies of endogenous opioids and their receptors related to behavior (Section 2), and the roles of these opioid peptides and receptors in pain and analgesia (Section 3); stress and social status (Section 4); tolerance and dependence (Section 5); learning and memory (Section 6); eating and drinking (Section 7); alcohol and drugs of abuse (Section 8); sexual activity and hormones, pregnancy, development and endocrinology (Section 9); mental illness and mood (Section 10); seizures and neurologic disorders (Section 11); electrical-related activity and neurophysiology (Section 12); general activity and locomotion (Section 13); gastrointestinal, renal and hepatic functions (Section 14); cardiovascular responses (Section 15); respiration (Section 16); and immunological responses (Section 17).