定量毛果芸香碱电渗汗液收集和检查方法的建立

定量毛果芸香碱电渗汗液收集和检查方法的建立陈柏华，张思仲，杨元囊性纤维化（ｃｙｓｔｉｃｆｉｂｒｏｓｉｓ，ＣＦ）是白种人中最常见的常染色体隐性遗传病，其发病率有明显的种族差异，在活产儿中发病率可高达１／２０００。我国虽已有散发病例报告，但发病率尚未确定...     

噻吗心安和溴莫尼定滴眼液对兔眼表组织的影响

目的 探讨降眼压药物噻吗心安滴眼液和溴莫尼定滴眼液及各自的防腐剂硫柳汞和氯化笨甲烃胺(BAC)对兔眼表组织的影响.方法 15只新西兰大白兔,随机分为对照组(3只)、噻吗心安组(T组,6只)和溴莫尼定组(B组,6只).实验兔右眼使用噻吗心安或溴莫尼定滴眼液,左眼使用对应的防腐剂硫柳求和BAC,每日2次,连续用药30 d.取球结膜组织进行HE染色并计数结膜上皮层炎性细胞数;角膜行扫描电镜检测并进行上皮损害分级评分.结果 HE染色示对照组兔球结膜上皮细胞排列整齐,用药后各组结膜上皮细胞层数增多,排列较对照组紊乱.与对照组比较,噻吗心安滴眼液、溴莫尼定滴眼液、防腐剂硫柳汞和BAC均可导致球结膜上皮层炎性细胞浸润增多(55.8+6.9,71.0±8.8,56.0±7.1,62.5±7.1比32.3±8.8,均P<0.05).角膜扫描电镜示除对照组外其他各组角膜上皮细胞表面的微绒毛丢失、细胞空洞和暗细胞增加;各实验组角膜上皮损害评分均高于对照组(P<0.05).溴莫尼定滴眼液引起的损害较噻吗心安滴眼液明显(P<0.05),但两种滴眼液和对应的防腐剂对左右两眼的损害差异无统计学意义(P>0.05).结论 噻吗心安、溴莫尼定滴眼液及防腐剂硫柳汞和BAC使用1个月后可导致兔球结膜炎性细胞增加和角膜上皮细胞损伤;防腐剂是引起眼表损伤的主要原因.溴莫尼定滴眼液对兔眼表的损伤较噻吗心安滴眼液严重.     

槐胺碱对细胞免疫的影响

槐胺碱对细胞免疫的影响张珮，金英，李淑华锦州医学院微生物学教研室锦州１２１００１槐胺碱（ＳＡ）系从豆科槐属植物苦豆子中提取的一种生物碱，曾有报道槐胺碱有抑制抗体产生的作用［１］。我们观察了槐胺碱对细胞免疫功能的影响。１材料和方法槐胺碱为宁夏盐池制药厂...     

我国囊性纤维化病的初步研究──282名儿童汗液钠离子浓度调查

采用定量毛果芸香碱电渗汗液收集和检查方法，收集了２月～１４岁的２８２例正常和有类似囊性纤维化病症状儿童的汗液，检查了他们的汗液钠离子浓度。结果表明，汗液钠离子浓度与收集部位、年龄、性别等无显著相关；我国儿童汗液钠离子浓度平均值为２４．０±８．６６ｍｍｏｌ／Ｌ，与白种人儿童无显著差异；未发现钠离子浓度超出正常范围者，就我国囊性纤维化病的发病率及其低下的原因进行了分析，并对该病的汗液检查方法和临床诊断应注意的问题进行了讨论。     

下丘注射去甲肾上腺素和毛果芸香碱对大鼠听源性癫痫发作的影响

双侧下丘注射NE或阿托品后,大鼠听源性癫痫发作(audiogenic seizure, AS)显著减弱。注射NE前5分钟,先在双侧下丘注射α_2-NE能受体阻断剂盐酸育亨宾(4μg),1小时后,NE对AS的抑制作用被完全逆转,而未预先注射育亨宾的大鼠的AS易感性仍显著低下。双侧下丘注射毒蕈碱样受体的激动剂毛果芸香碱(10μg)后,AS易感品系中不发作的大鼠表现出AS易感性。 实验结果提示:下丘NE水平的提高减弱AS,NE可能是作用于下丘内与外周α_2-NE能受体相似的受体而抑制AS的。下丘毒蕈碱样受体的激活对AS起促进作用。     

毛果甘青乌头中生物碱成分的提取和鉴定

目的：分离纯化鉴定毛果甘青乌头中的生物碱成分。方法：药材室温下自然烘干后,粉粹,用酸提碱沉法提取总生物碱,然后用硅胶柱层析法分离得到单体生物碱,利用化学法和光谱法鉴定其结构。结果：分离鉴定得到了10个化合物的结构,其中有六个内酯型C19二萜生物碱、四个C20二萜生物碱。结论：得到了10个纯的单体生物碱。     

小檗碱对小鼠淋巴细胞体外增殖和细胞周期的影响

目的 :分析小檗碱 (Ber)对小鼠淋巴细胞体外增殖和细胞周期的影响 ,探讨其免疫抑制作用的机制。方法 :分离小鼠淋巴结细胞 ,以CFSE染色 ,加入多克隆刺激剂刀豆蛋白 (ConA)或者佛波醇酯 (PDB)和离子霉素 (Ion)进行刺激 ,以流式细胞仪分析小檗碱对淋巴细胞增殖率的影响 ;以WST 1检测淋巴细胞的增殖情况 ;用碘化丙锭 (PI)染色分析细胞周期分布 ;细胞凋亡以Annexin V染色进行分析。结果 :CFSE染色分析显示 ,当浓度为 10 0、30和 10 μmol L时 ,Ber无论对ConA或PDB Ion诱导的小鼠淋巴细胞增殖 ,都具有明显的抑制作用 (P <0 0 5 )。以WST 1分析细胞增殖情况 ,也获得相似的结果。对细胞周期分析表明 ,随着小檗碱浓度的增加 ,处于亚二倍体峰的细胞数增加 ,处于S和G2 M期的细胞数则减少 ,而G0 G1 期的细胞数基本不变。Annexin V染色法结果表明 ,30 μmol LBer组Annexin V单阳性区域的百分率为 ( 7 13± 1 0 8) % ,与PDB Ion阴性组的百分率 ( 2 4 9± 0 2 5 ) %比较有显著的差异 (P <0 0 1)。结论 :小檗碱对小鼠淋巴细胞的增殖有明显抑制作用 ,它能抑制淋巴细胞进入细胞分裂周期 ,这种抑制作用表现出明显的周期特异性。同时 ,Ber可以诱导体外培养的小鼠淋巴细胞发生凋亡。     

川芎嗪对兔眼高压微循环影响的研究

目的 探讨川芎嗪对兔眼持续高压微循环的影响。方法 对36只新西兰白兔前房注入α－糜蛋白酶制成持续性高眼压模型,随机等分为高眼压对照组和川芎嗪治疗组。治疗组用于高眼压持续第7d始肌注川芎嗪,观察不同时间两组动物球结膜微循环变化。结果 与高眼压对照组相比,川嗪组球膜微循环障碍明显减轻。结论 川芎嗪可改善微循环状况从而对高眼压兔眼视神经起到保护作用。     

曲伏前列素联合布林佐胺治疗原发性开角型青光眼的临床疗效及安全性

目的 探讨曲伏前列素联合布林佐胺治疗原发性开角型青光眼（POAG）的临床疗效及安全性。方法 选取2016年1月~2018年12月我院收治的POAG患者112例,采用随机数字表法分为对照组和观察组,每组56例。对照组给予曲伏前列素滴眼液治疗,观察组在对照组基础上给予布林佐胺滴眼液。比较两组平均眼压、昼夜眼压差、临床疗效及不良反应。结果 治疗后,观察组平均眼压和昼夜眼压差均低于对照组[（16.26±2.14）mmHg vs（17.70±2.66）mmHg]、[（1.89±0.52）mmHg vs（2.33±0.56）mmHg],差异有统计学意义（P＜0.05）。观察组总有效率为96.43%,高于对照组的75.00%,差异有统计学意义（P＜0.05）。两组不良反应发生率比较,差异无统计学意义（P＞0.05）。结论 曲伏前列素联合布林佐胺治疗POAG临床疗效确切,可有效降低眼压,稳定昼夜眼压,不增加不良反应。     

柔红霉素、骆驼蓬总碱对兔视网膜的毒性观察

目的 探讨药物柔红霉素(daunorubicin,DNR)、骆驼蓬总碱(total alkaloid Of Harmaline,TAH)眼内注射防治后发性白内障对视网膜的毒性作用.方法 于兔右眼晶体囊摘除(extracapsular lens extraction,ECLE)术中分别前房内注射0.2mg/mlTAH和0.2mg/ml DNR溶液0.1ml,通过眼底镜检查、视网膜电图及眼组织病理学等研究对象兔眼视网膜的影响.结果 对照眼与15只注入TAH眼眼底无明显的病理变化;TAH眼手术前后视网膜电图b波的振幅和潜时与对照眼无差别.而10只注入DNR眼出现严重的内皮性角膜水肿混浊,前房大量纤维索性渗出特等明显的炎性反应眼底及视网膜电图无法检查.组织病理学检查显示TAH组眼前节结构及视网膜各层结构正常,细胞排列规律;透射电镜检查TAH眼的视网膜感光细胞外节盘膜结构清晰.排列整齐.视网膜细胞排列紊乱,部分明显呈核固缩,表现出明显的毒性反应.结论 TAH眼内注射对兔视网膜的毒性较小,有可能用于后发性白内障的防治研究,而DNR对兔视网膜及其它组织表现出明显毒性,应进一步研究其使用的安全剂量和剂型.     

Effects of the muscarinic agonist pilocarpine on vigilance states and locomotor activity in ring doves

Cholinergic systems play a significant role in regulating a variety of behavioral functions in mammals and birds. The aim of this work is to study the effects of the muscarinic agonist pilocarpine on behavioral states by visual inspection and electroencephalographic recording; also, locomotor activity was continuously recorded by infrared interruption system in ring doves. The current results in birds demonstrated that the muscarinic agonist pilocarpine (1 and 3 mg/kg, i.p.) primarily induced theta activity in addition to promote passive waking, while diminished active waking, the EEG slow wave rhythm and REM sleep in ring doves. The locomotor activity recorded continuously in ring doves diminished after pilocarpine treatment, which was in good agreement with the observed reduction of active waking derived of the EEG study. Altogether, the current results are similar to the effects of pilocarpine previously reported in mammals. In conclusion, hippocampal theta rhythm in birds suggests that this rhythm is an ancestral property of hippocampal function and similar cholinergic mechanisms regulate vigilance states and theta generation in mammals and birds.     

A gelatin-g-poly(N-isopropylacrylamide) biodegradable in situ gelling delivery system for the intracameral administration of pilocarpine

In this study, the aminated gelatin was grafted with carboxylic end-capped poly(N-isopropylacrylamide) (PN) via a carbodiimide-mediated coupling reaction to fabricate biodegradable in situ forming delivery systems for intracameral administration of antiglaucoma medications. The chemical structure of the graft copolymers (GN) was confirmed by Fourier transform infrared (FTIR) spectroscopy. When the feed molar ratio of NH₂/COOH was 0.36, the grafting ratio, efficiency and degree of grafting, and weight ratio of PN to aminated gelatin was 25.6, 18.6%, 52.6%, and 1.9, respectively. As compared to PN, the GN samples possessed better thermal gelation ability and adherence, indicating remarkable phase transition properties. Under gelatinase degradation, the remaining weight of GN was significantly lower than those of PN at each time point from 8 h to 4 weeks. Cytocompatibility studies showed that the culture of anterior segment cells with both in situ forming gels does not affect proliferation and has little effect on inflammation. Higher encapsulation efficiency (∼62%) and cumulative release (∼95%) were achieved for GN vehicles, which was attributed to initial fast temperature triggered capture of pilocarpine and subsequent progressive degradation of gelatin network. In a rabbit glaucoma model, the performance of delivery carriers was evaluated by biomicroscopy, intraocular pressure (IOP), and pupil size change. Intracameral administration of pilocarpine using GN was found to be more effective than other methods such as instillation of eye drop and injection of free drug or PN containing drug in improving ocular bioavailability and extending the pharmacological responses (i.e., miosis and IOP lowering effect and preservation of corneal endothelial cell density).     

Anticonvulsant effect of phytol in a pilocarpine model in mice

The present study investigated the effects of phytol in pilocarpine-induced seizures. The latency for development of convulsions and mortality rate was recorded in this model using mice. The results revealed that phytol (25, 50 and 75 mg/kg, i.p.) increased latency to first seizure and decreased percentage of these seizures. Moreover, phytol also protected the animals against status epilepticus induced by pilocarpine, and decreased the mortality rate. Mice treated with pilocarpine (n = 24) presented 100% of mortality during the first hour of observation. In turn, phytol-pretreated animals within 30 min before the administration of pilocarpine (400 mg/kg) remained alive during the first hour of observation. On the other hand, 6–8 h after administration of pilocarpine it was observed that 10 (41.66%), 8 (33.33%) and 4 (16.66%) animals died (respectively). Thus, the pretreatment with phytol was able to block mortality rate during the first hour in acute phase of seizures, and significantly reduced this rate in a dose-dependent manner (p < 0.05), suggesting an anticonvulsant effect. In addition, none of the phytol effects was blocked by pre-treatment with flumazenil, an antagonist of benzodiazepine receptors. In conclusion, phytol exhibits anticonvulsant activity by modulating of neurotransmitter systems, but further investigations are in progress to confirm this pharmacological property.     

Vesicular acetylcholine transporter knock-down mice are more susceptible to pilocarpine induced status epilepticus

The pilocarpine (PILO) animal model of Temporal Lobe Epilepsy (TLE) portrays the most common changes in hippocampal circuitry found in human TLE. The acute cholinergic insult induces status epilepticus (SE), which triggers an overwhelming set of plastic events that result on late spontaneous recurrent limbic seizures. It has been suggested that the cholinergic system plays an important role in the synchronization required for ictogenesis. We took advantage of a knock-down animal model for the vesicular acetylcholine transporter (VAChT KD) to investigate seizure genesis in a model of cholinergic dysfunction. We induced SE in VAChT KD and wild-type (WT) mice by a single intraperitoneal injection of PILO in order to evaluate susceptibility to seizures. Video-EEG recordings evaluated epileptiform activity and ictal behavior onset. The hypothesis tested is that innate cholinergic hypofunction could result in increased susceptibility to PILO. VAChT KD^HOM mice showed shorter latency for the first epileptiform discharge and for the first seizure episode, when compared to other groups. The duration of these seizure episodes, however, were not statistically different among experimental groups. On the other hand, VAChT KD^HOM had the shortest latency to isoelectric EEG, when compared to WT and KD^HET. Our results indicate that a reduction of brain VAChT protein to the levels found in VAChT KD^HOM mice alters the epileptic response to PILO. Thus, fine-tuning modulation of cholinergic tone can affect the susceptibility of epileptic responses to pilocarpine.     

Effects of the vitamin E in catalase activities in hippocampus after status epilepticus induced by pilocarpine in Wistar rats

Experimental manipulations suggest that in vivo administration of exogenous antioxidants agents decreases the concentration of free radical in the brain. Neurochemical studies have proposed a role for catalase in brain mechanisms responsible by development to status epilepticus (SE) induced by pilocarpine. The present study was aimed at was investigating the changes in catalase activities after pilocarpine-induced SE. Animals were treated with vitamin E (VIT E) 200 mg/kg (intraperitoneally (i.p.)) and, 30 min later, they received pilocarpine hydrochloride, 400 mg/kg, subcutaneous (s.c.) (P400). Other three groups received VIT E (200 mg/kg, i.p.), pilocarpine (400 mg/kg, s.c.) or 0.9% NaCl (control) alone. Animals were closely observed for behavioral changes, tremors, stereotyped movements, seizures, SE and death, for 24 h following the pilocarpine injection. The brains were dissected after decapitation. The results have shown that pilocarpine administration and resulting SE produced a significant increase in hippocampal catalase activity of (88%). In the group pre-treated which VIT E in hippocampal catalase activity was increase of 67% and 214% when compared with P400 and control group, respectively. Our results demonstrated a direct evidence of an increase in the activity of the hippocampal catalase of rat adults during seizure activity and after the pre-treated which VIT E that could be responsible by regulation of free radical levels during the establishment of SE.     

早期安定干预对匹罗卡品诱导颞叶癫痫小鼠学习记忆能力变化的影响

目的:研究早期使用安定麻醉对匹罗卡品(PILO)诱导的颞叶癫痫小鼠的学习记忆能力的变化,从行为学角度评价抑制点燃效应对颞叶癫痫造模程度的影响。方法:腹腔注射PILO建立颞叶癫痫小鼠模型,分PILO癫痫组(A组)、安定麻醉条件下PILO癫痫组(B组)、安定麻醉对照组(C组),正常对照组(D组),对照组腹腔注射与PILO等量的生理盐水。各组小鼠进行Morris水迷宫学习记忆实验,测其寻找到隐藏平台所需潜伏期。结果:Morris水迷宫实验中,A组癫痫小鼠逃避潜伏期时间明显延长(P<0.05),B组癫痫小鼠第一次癫痫发作程度有大幅度的减轻,逃避潜伏期时间延长,但延长时间较正常情况下点燃癫痫小鼠低。结论:早期使用安定麻醉可明显改善颞叶癫闲小鼠学习记忆能力。     

Neuropeptide-Y immunoreactivity in the pilocarpine model of temporal lobe epilepsy

 Neuropeptide-Y (NPY) is expressed by granule cells and mossy fibres of the hippocampal dentate gyrus during experimental temporal lobe epilepsy (TLE). This expression may represent an endogenous damping mechanism since NPY has been shown to block seizure-like events following high-frequency stimulation in hippocampal slices. The pilocarpine (PILO) model of epilepsy is characterized by an acute period of status epilepticus followed by spontaneous recurrent seizures and related brain damage. We report peroxidase-antiperoxidase immunostaining for NPY in several brain regions in this model. PILO-injected animals exhibited NPY immunoreactivity in the region of the mossy fibre terminals, in the dentate gyrus inner molecular layer and, in a few cases, within presumed granule cells. NPY immunoreactivity was also dramatically changed in the entorhinal cortex, amygdala and sensorimotor areas. In addition, PILO injected animals exhibited a reduction in the number of NPY-immunoreactive interneurons compared with controls. The results demonstrate that changes in NPY expression, including expression in the granule cells and mossy fibres and the loss of vulnerable NPY neurons, are present in the PILO model of TLE. However, the significance of this changed synthesis of NPY remains to be determined. Received: 19 August 1996 / Accepted: 21 March 1997