高效毛细管电泳法分离多巴胺和5—羟色胺

建立毛细管电泳分离分析多巴胺和5－羟色胺的方法。采用自由区带电泳法（CZE），40mmol/L硼带缓冲液20PSI气压进样5s，定电流75μA分离10min，二级管阵列PDA检测器检测，应用200nm检测波长，结果显示两种物质完全分离。     

高效毛细管电泳法分离肾上腺素和去甲肾上腺素

利用毛细管电泳法分离肾上腺素，去甲肾上腺素，采用自由区带电泳法(CZE)，40mmol／L硼砂缓冲液20PSI气压进样5s,等电压20KV分离12min，应用200nm波长检测，分离温度25℃。结果显示两种物质完全分离。     

高效毛细管电泳法分离肾上腺素和去甲肾上腺素

利用毛细管电泳法分离肾上腺素，去甲肾上腺素，采用自由区带电泳法(CZE)，40mmol/L硼砂缓冲液20PSI气压进样5s，等电压20KV分离12min，应用200nm波长检测，分离温度25℃。结果显示两种物质完全分离。     

高效毛细管电泳法分析肝癌单克隆抗体的纯度

高效毛细管电泳法（ｈｉｇｈｐｅｒｆｏｒｍａｎｃｅｃａｐｉｌｌａｒｙｅｌｅｃｔｒｏｐｈｏｒｅｓｉｓ,ＨＰＣＥ）是８０年代初创立、近几年发展最快的一种强有力的分析分离技术［１］。它不仅综合了高效液相色谱和凝胶电泳的共同优点,而且具有其自己的特点：高效快速、高分辨率、高灵敏度、样品用量少、分离模式多、重复性好、自动化程度高。只要淌度稍有差异即可达到分离。目前,ＨＰＣＥ在蛋白质、多肽、氨基酸、核酸,甚至在离子分析方面的应用研究十分活跃。本文运用ＨＰＣＥ对肝癌单克隆抗体（ｍＡｂ）ＨＡｂ１８的纯度进行了分析…     

高效毛细管电泳法分析rhIL—2的纯度

重组人白细胞介素 2 ( rh IL - 2 )是利用基因工程技术 ,从大肠杆菌中获得的。 rh IL- 2能抑制肿瘤细胞的增殖 ,是肿瘤过继免疫治疗中应用最广泛 ,研究得最多的细胞因子 [1 ] 。作为生物制剂 ,在药品的质量监控中 ,其纯度分析又是非常重要的指标之一。目前 ,常用的纯度鉴定方法有凝胶电泳和高效液相色谱 ( HPL C)。与 HPL C相比 ,高效毛细管电泳( HPCE)具有更高的分辨率和灵敏度 ,且同时具有快效、高效、自动化、上样量少等特点 [2 ] 。本文运用 HPCE对 rh IL - 2进行纯度分析 ,建立了快速简便的测定方法。1　材料与方法1.1　仪器…     

毛细管电泳的最新进展

毛细管电泳是近年发展最快的分离分析技术之一。它具有高灵敏度、高分辨率、高速度等优点 ,广泛应用于各个领域。随着毛细管电泳技术的不断发展 ,逐渐出现了非水毛细管电泳 ,毛细管阵列电泳 ,毛细管电泳免疫分析 ,毛细管电色谱手性拆分等分支     

非水毛细管电泳法研究进展

非水毛细管电泳作为毛细管电泳法的重要分支已成为研究热点,近年来被广泛应用。综述了非水溶剂的特性及其分子间的相互作用。总结了非水毛细管电泳法中调节和增强选择性的方法和常用检测方法,以及近年来非水毛细管电泳法应用于生物样本代谢产物及药物等方面的研究进展,并展望了其广泛的应用前景。     

反相高效液相色谱法测定血清中的5-羟色胺

建立了一种简便、快速测定血清中5-羟色胺的反相高效液相分析方法。采用的色谱柱为Bondapak C18不锈钢柱(5μm,250mm×4.6mmi.d.),流动相为:甲醇—水—乙酸(70:30:0.1 V/V),流速:1ml/min,荧光监测器,激发波长278nm,发射波长333 nm。结果表明:该方法线性关系良好(r=0.99992),5-羟色胺测定的线性范围为0.10~2.00μg/mL,最低检测限为0.02μg/mL(S/N=3)。5-羟色胺在血清样品中的加标回收率为92.37%~100.12%。该测定方法结果准确,可用于临床研究。     

加昧逍遥散对肝癌大鼠脑内多巴胺和5-羟色胺水平的影响

目的:探讨中药方剂加味逍遥散(JWXYP)对肝郁脾虚型肝癌大鼠脑内单胺类神经递质多巴胺(DA)和5-羟色胺(5-HT)水平的影响。方法:将36只SD大鼠随机分成对照组(control)、肝癌模型组(HCC)、加味逍遥散治疗组(JWXYP)。通过二乙基亚硝胺(DNE)结合不可预见性应激刺激(CUMS)构建肝郁脾虚型肝癌大鼠模型,给子中药加味逍遥散治疗4周。通过症状和体征观察各组大鼠症候变化,利用HE染色观察大鼠肝脏病理学形态改变,高效液相-电化学检测法检测额叶皮质内DA和5-HT水平。结果:与对照组比较,HCC组大鼠病理表现为肝小叶结构破坏,增生细胞有异型性表现,可见双核细胞、病理性核分裂象等,症候评分显示符合肝郁脾虚型症候表现,脑内DA和5-HT显著下降。经过JWXYP治疗,大鼠肝组织病理恶性程度较轻,症候明显改善,脑内DA和5-HT显著增加。结论:加味逍遥散能够改善肝郁脾虚型肝癌模型大鼠症候,其机制可能与增加脑内DA和5-HT有关。     

毛细管电泳在PCR产物及核糖核酸分离分析中的应用

毛细管电泳具有快速、微量、分辨率高、重复性好及易于定量，易于自动化的特点，可高效分离，分析ＲＮＡ，ｓｓＤＮＡ、ｄｓＤＮＡ，从而提供了一种ＰＣＲ产物自动化检测系统，已广泛用于核酸定量、细菌、病毒基因的鉴定，突变基因的检测，随着ＣＥ研究的进一步发展，可望成为传染病学，肿瘤学，分子遗传学等研究领域中一种不可或缺少的研究手段。     

Direct monitoring of dopamine and 5-HT release in substantia nigra and ventral tegmental area in vitro

Fast-scan cyclic voltammetry with carbon fibre microelectrodes was used to detect endogenous dopamine (DA) and 5-hydroxytryptamine (5-HT) release from three distinct regions of guinea-pig mid-brain in vitro: rostral and caudal substantia nigra (SN) and the ventral tegmental area (VTA). Previous electrophysiological studies have demonstrated that cells of the caudal SN and the VTA have similar characteristics, whereas cells in the rostral SN have distinctly different properties. In the present study, we confirmed that each region has tyrosine hydroxylase-positive neurons and determined, using high-performance liquid chromatography, that DA levels were similar in rostral and caudal SN, but lower in SN than in VTA. In each region, application of veratrine, which was shown by intracellular recordings to have a reversible depolarising action, evoked a signal attributable to DA and distinguishable from that of 5-HT. Release signals were monitored every 250 ms with a spatial resolution of less than 50 m. DA release was calcium-dependent and was not detectable in a catecholamine-poor area such as the cerebellum, or in mid-brain tissue pre-treated with reserpine. Within the normal mid-brain, the amount of DA released was correlated with tissue content in that it was higher in the VTA than in either region of SN. It is concluded that DA released from somato-dendritic parts of mid-brain neurons exhibits site-specific variation. This is the first report of direct monitoring of DA and 5-HT relase from these regions with in situ electrodes and demonstrates the utility of fast-scan cyclic voltammetry to investigate the mechanisms and possible non-classical functions of somato-dendritic DA release.     

Direct monitoring of dopamine and 5-HT release in substantia nigra and ventral tegmental area in vitro

Fast-scan cyclic voltammetry with carbon fibre microelectrodes was used to detect endogenous dopamine (DA) and 5-hydroxytryptamine (5-HT) release from three distinct regions of guinea-pig mid-brain in vitro : rostral and caudal substantia nigra (SN) and the ventral tegmental area (VTA). Previous electrophysiological studies have demonstrated that cells of the caudal SN and the VTA have similar characteristics, whereas cells in the rostral SN have distinctly different properties. In the present study, we confirmed that each region has tyrosine hydroxylase-positive neurons and determined, using high-performance liquid chromatography, that DA levels were similar in rostral and caudal SN, but lower in SN than in VTA. In each region, application of veratrine, which was shown by intracellular recordings to have a reversible depolarising action, evoked a signal attributable to DA and distinguishable from that of 5-HT. Release signals were monitored every 250 ms with a spatial resolution of less than 50 μm. DA release was calcium-dependent and was not detectable in a catecholamine-poor area such as the cerebellum, or in mid-brain tissue pre-treated with reserpine. Within the normal mid-brain, the amount of DA released was correlated with tissue content in that it was higher in the VTA than in either region of SN. It is concluded that DA released from somato-dendritic parts of mid-brain neurons exhibits site-specific variation. This is the first report of direct monitoring of DA and 5-HT relase from these regions with in situ electrodes and demonstrates the utility of fast-scan cyclic voltammetry to investigate the mechanisms and possible non-classical functions of somato-dendritic DA release.     

集成毛细管电泳芯片研究进展

集成毛细管电泳芯片是一种新型的微全分析系统,它具有样品用量少、分析速度快、体积小便于携带、成本低等优点。本文综述了该芯片的产生和发展过程以及芯片的结构、进样方式、检测方法等方面的进展和存在的问题;简述了其主要应用领域并展望了其应用前景。     

人参二醇组皂甙对失血性休克犬血清5-HT的影响及对血小板的保护作用

健康成年杂种犬29只,分为失血性休克人参预治疗组(HSG)10只,失血性休克地塞米松预治疗组(HSD)10只,失血性休克组(HS)9只。失血前1小时,HSG组静注人参二醇组皂甙(25mg/kg),HSD组肌注地塞米松(1mg/kg)。在实验过程中经放、输血,使血压维持于5.3kPa(40mmHg)。结果表明,(1)扫描电镜显示,人参二醇组皂甙与地塞米松对血小板形态均有明显保护作用,并抑制其聚集;(2)血清5-HT含量,HS组随失血时间延长逐渐增高。HSG组失血后第1、3小时虽有增高,但第4、5小时则逐渐回降到正常水平,HSD组失血前后均无显著改变。5-HIAA,HS组失血后3～5小时显著增加,HSG和HSD组失血前后比较均无显著改变。提示人参二醇组皂甙和地塞米松对失血性休克时5-HT和5-HIAA的增加有抑制作用。     

Effect of the 5-HT4 receptor and serotonin transporter on visceral hypersensitivity in rats

Visceral hypersensitivity plays an important role in motor and sensory abnormalities associated with irritable bowel syndrome, but the underlying mechanisms are not fully understood. The present study was designed to evaluate the expression of the 5-HT₄ receptor and the serotonin transporter (SERT) as well as their roles in chronic visceral hypersensitivity using a rat model. Neonatal male Sprague-Dawley rats received intracolonic injections of 0.5% acetic acid (0.3-0.5 mL at different times) between postnatal days 8 and 21 to establish an animal model of visceral hypersensitivity. On day 43, the threshold intensity for a visually identifiable contraction of the abdominal wall and body arching were recorded during rectal distention. Histological evaluation and the myeloperoxidase activity assay were performed to determine the severity of inflammation. The 5-HT₄ receptor and SERT expression of the ascending colon were monitored using immunohistochemistry and Western blot analyses; the plasma 5-HT levels were measured using an ELISA method. As expected, transient colonic irritation at the neonatal stage led to visceral hypersensitivity, but no mucosal inflammation was later detected during adulthood. Using this model, we found reduced SERT expression (0.298 ± 0.038 vs 0.634 ± 0.200, P < 0.05) and increased 5-HT₄ receptor expression (0.308 ± 0.017 vs 0.298 ± 0.021, P < 0.05). Treatment with fluoxetine (10 mg·kg⁻¹·day⁻¹, days 36-42), tegaserod (1 mg·kg⁻¹·day⁻¹, day 43), or the combination of both, reduced visceral hypersensitivity and plasma 5-HT levels. Fluoxetine treatment increased 5-HT₄ receptor expression (0.322 ± 0.020 vs 0.308 ± 0.017, P < 0.01) but not SERT expression (0.219 ± 0.039 vs 0.298 ± 0.038, P = 0.654). These results indicate that both the 5-HT₄ receptor and SERT play a role in the pathogenesis of visceral hypersensitivity, and its mechanism may be involved in the local 5-HT level.     

Serotonin and psychostimulant addiction: focus on 5-HT1A-receptors

Serotonin(1A)-receptors (5-HT(1A)-Rs) are important components of the 5-HT system in the brain. As somatodendritic autoreceptors they control the activity of 5-HT neurons, and, as postsynaptic receptors, the activity in terminal areas. Cocaine (COC), amphetamine (AMPH), methamphetamine (METH) and 3,4-methylenedioxymethamphetamine ("Ecstasy", MDMA) are psychostimulant drugs that can lead to addiction-related behavior in humans and in animals. At the neurochemical level, these psychostimulant drugs interact with monoamine transporters and increase extracellular 5-HT, dopamine and noradrenalin activity in the brain. The increase in 5-HT, which, in addition to dopamine, is a core mechanism of action for drug addiction, hyperactivates 5-HT(1A)-Rs. Here, we first review the role of the various 5-HT(1A)-R populations in spontaneous behavior to provide a background to elucidate the contribution of the 5-HT(1A)-Rs to the organization of psychostimulant-induced addiction behavior. The progress achieved in this field shows the fundamental contribution of brain 5-HT(1A)-Rs to virtually all behaviors associated with psychostimulant addiction. Importantly, the contribution of pre- and postsynaptic 5-HT(1A)-Rs can be dissociated and frequently act in opposite directions. We conclude that 5-HT(1A)-autoreceptors mainly facilitate psychostimulant addiction-related behaviors by a limitation of the 5-HT response in terminal areas. Postsynaptic 5-HT(1A)-Rs, in contrast, predominantly inhibit the expression of various addiction-related behaviors directly. In addition, they may also influence the local 5-HT response by feedback mechanisms. The reviewed findings do not only show a crucial role of 5-HT(1A)-Rs in the control of brain 5-HT activity and spontaneous behavior, but also their complex role in the regulation of the psychostimulant-induced 5-HT response and subsequent addiction-related behaviors.     

The Effects of Hypothyroidism on 5-HT1A and 5-HT2A Receptors and the Serotonin Transporter Protein in the Rat Brain

The effects of hypothyroidism on 5-HT_1A and 5-HT_2A receptors and the serotonin transporter protein were studied in thyroidectomized male Wistar rats in two experimental groups: 1) animals kept on an iodine-free diet (hypothyroid rats) and 2) animals kept on thyroxine (15 g/kg) for 21 days (giving normal thyroid hormone levels, euthyroid animals). Sham-operated rats served as controls. Binding of [³H]8-OH-DPAT with 5-HT_1A receptors and [³H]citalopram with the transporter protein in the hippocampus and midbrain showed no changes in hypothyroid rats as compared with controls. Conversely, there were significant decreases in [³H]ketanserin binding to 5-HT_2A receptors in the frontal cortex in hypothyroid rats as compared with controls; this decrease was reversed by thyroxine treatment. Thus, losses of cortical 5-HT_2A receptors appears to be the main consequence of hypothyroidism at the level of the serotonin system of the brain.     

Effects of the 5-HT4 receptor agonist RS67333 and paroxetine on hippocampal extracellular 5-HT levels

The 5-HT₄ receptor modulates activity of serotonergic neurons and is a new potential target for antidepressant treatment. This microdialysis study evaluated the effect of the 5-HT₄ receptor agonist, RS67333, on extracellular serotonin (5-hydroxytryptamine, 5-HT) and 5-HIAA levels in rat ventral hippocampus during chloral hydrate anaesthesia, and explored the ability of RS67333 to augment the effect of the selective serotonin reuptake inhibitor paroxetine. The effect of RS67333 was examined after acute and subchronic (3 days) administration. Acute RS67333 (1.5 mg/kg i.v.) had no effect on extracellular 5-HT or 5-HIAA levels, while acute paroxetine (0.5 mg/kg i.v.) increased 5-HT levels by 299 ± 16% and decreased 5-HIAA levels by 25 ± 4%. Administration of RS67333 80 min after paroxetine caused an additional transient increase in 5-HT levels (to 398 ± 52% of baseline). Subchronic RS67333 administration (1.5 mg/kg i.p.) increased basal 5-HT levels by 73 ± 15% and decreased 5-HIAA levels by 27 ± 13%. In conclusion, the 5-HT₄ receptor agonist RS67333 augmented the acute effect of paroxetine on extracellular 5-HT levels in the ventral hippocampus, and after 3 days increased basal hippocampal 5-HT levels.