司来吉兰对帕金森病模型大鼠结肠α-Syn Bcl-2表达的影响

目的观察司来吉兰对帕金森病模型大鼠结肠中α-Syn及Bcl-2表达的影响,探讨司来吉兰对帕金森病消化系统功能紊乱的可能机制。方法 60只健康雄性SD大鼠随机分为对照组、PD组以及治疗组,每组均设4d及8d2个时间点。采用颈背部皮下注射鱼藤酮制备帕金森病模型大鼠成功后,治疗组每日灌胃司来吉兰0.5mg/kg,直至试验时间点。对照组及PD组每日均连续灌胃等体积生理盐水至试验时间点。应用免疫组化法S-P法以及Western blotting检测α-Syn及Bcl-2在3组大鼠结肠组织中的表达情况。结果造模成功后,免疫组化及Western blotting结果均显示,PD组大鼠结肠中的Bcl-2表达较对照组明显减低(P0.05);通过司来吉兰治疗后,治疗组Bcl-2表达显著高于PD组,但仍低于对照组(P0.05),治疗8d时较治疗4d时,Bcl-2表达显著增强(P0.05)。α-Syn在PD组中的表达较对照组以及治疗组明显增多(P0.05),且治疗组中α-Syn表达较对照组增多(P0.05);且治疗8d时较治疗4d时α-Syn表达减低(P0.05)。结论帕金森大鼠结肠组织中的Bcl-2表达降低,而α-Syn表达增加。司来吉兰能缓解PD后Bcl-2的降低以及α-Syn的升高,能有效缓解帕金森患者的肠道功能紊乱。     

司来吉兰联合左旋多巴对帕金森病模型大鼠结肠TH及nNOS表达的影响

目的观察司来吉兰联合左旋多巴对帕金森病模型大鼠结肠中TH及nNOS表达的影响,探讨司来吉兰联合左旋多巴对帕金森病消化系统功能紊乱的可能机制。方法 72只健康雄性SD大鼠随机分为对照组、模型组及联合治疗组,每组均设4d及8d2个时间点。采用颈背部皮下注射鱼藤酮制备帕金森病模型大鼠,模型制备成功后,联合治疗组每日灌胃1次司来吉兰0.5mg/kg及2次左旋多巴15mg/kg,直至试验时间点。对照组及模型组每只每日均连续灌胃等体积生理盐水至试验时间点。应用免疫组化法S-P法以及Western blotting检测TH及nNOS在对照组、模型组以及联合治疗组大鼠结肠组织中的表达情况。结果造模成功后,免疫组化及Western blotting结果均显示,模型组大鼠结肠中的TH表达较对照组明显减低(P0.05);通过司来吉兰联合左旋多巴治疗后,联合治疗组TH表达显著高于模型组,但仍低于对照组(P0.05),治疗8d时较治疗4d时,TH表达无显著差异(P0.05)。nNOS在模型组中的表达较对照组以及联合治疗组明显增多(P0.05),且联合治疗组中nNOS表达较对照组增多(P0.05),治疗8d时较治疗4d时,nNOS表达无显著性差异(P0.05)。结论帕金森病大鼠结肠组织中的TH表达降低,而nNOS表达增加。司来吉兰联合左旋多巴能缓解PD后TH的降低以及nNOS的升高,能有效缓解帕金森大鼠的肠道功能紊乱。     

白芍总苷对EAE大鼠中枢神经系统炎症浸润细胞凋亡及Bcl-2、Bax表达的影响

目的 探讨白芍总苷（TGP）对实验性自身免疫性脑脊髓炎（EAE）大鼠中枢神经系统（CNS）炎症浸润细胞凋亡及Bcl-2、Bax表达的影响.方法 建立大鼠EAE模型,将大鼠随机分为正常对照组、模型组、TGP组、泼尼松组,TGP组免疫后第1天起每天经口灌服白芍总苷悬浊液0.2 g/kg,泼尼松组给予泼尼松,正常对照组、模型组给予同体积生理盐水溶液,第17天处死,病理切片观察CNS炎症细胞浸润情况,TUNEL法检测浸润细胞凋亡情况,免疫组化检测浸润细胞Bcl-2、Bax蛋白的表达.结果 泼尼松组、TGP组与模型组相比,中枢神经系统炎症浸润细胞的数目减少,凋亡率增高.TGP组与模型组相比,Bcl-2表达下降,Bax的表达上调,Bcl-2/Bax的比值下降.泼尼松组与模型组相比,Bcl-2/Bax的比值下降.结论 TGP能减轻EAE的病情,其机制可能是下调Bcl-2的表达,上调Bax蛋白的表达,降低Bcl-2/Bax比值,促进EAE大鼠CNS炎症浸润细胞的凋亡,减少CNS炎症细胞的浸润.     

醒脑静注射液对大鼠脑损伤后细胞凋亡及相关蛋白表达的影响

目的探讨醒脑静注射液对大鼠创伤性脑损伤(TBI)后细胞凋亡的影响及其机制。方法将大鼠随机分为假手术组、损伤对照组和醒脑静注射液治疗组,后两组再分为伤后6h、12h、24h、48h、72h和168h6个亚组,采用改进的Feeney法制作大鼠TBI模型,原位末端标记法(TUNEL)检测细胞凋亡,免疫组化法检测Bcl-2、Bax和Caspase-3的表达。结果治疗组大鼠脑细胞凋亡率及Bax、Caspase-3表达较损伤对照组显著降低(P<0.05),而Bcl-2表达明显升高(P<0.05)。结论醒脑静注射液对TBI的脑保护作用机制可能与干预伤后凋亡相关基因表达并减少神经细胞凋亡有关。     

拉莫三嗪对癫痫大鼠学习记忆及神经细胞凋亡的影响研究

目的 观察拉莫三嗪(lamotrigine,LTG)对戊四氮(PTZ)所致慢性癫痫大鼠学习记忆及神经细胞凋亡的影响,为LTG改善认知功能提供理论依据.方法 成年健康SD大鼠30只,随机分为对照组、模型组和LTG干预组各10只,采用35mg/kg PTZ溶液经腹腔注射制备癫痫模型.4w后对大鼠进行空间学习能力检测,应用免疫组化染色及原位细胞凋亡检测脑组织Bcl-2、Bax蛋白表达阳性细胞数及凋亡细胞数.结果 癫痫大鼠的学习记忆功能较正常大鼠明显下降(P＜0.05),LTG干预组海马区凋亡细胞明显减少(P＜0.01),Bcl-2蛋白阳性细胞数显著增加(P＜0.01),Bax蛋白阳性细胞数显著减少(P＜0.05).结论癫痫后大鼠的学习记忆功能受损,应用LTG可改善这一损害,可能与其能上调Bcl-2蛋白表达、下调Bax蛋白表达有关.     

丁苯酞对溴化乙锭诱导的脱髓鞘大鼠胼胝体Bcl-2和Bax表达的影响

目的观察丁苯酞(Dl-3-n-butylphthalide,NBP)对溴化乙锭(ehidium bromide,EB)诱导的脱髓鞘大鼠脑胼胝体凋亡因子B淋巴细胞瘤2基因(Bcl-2)和Bcl-2相关X蛋白(Bax)表达水平的影响,并探讨NBP对脱髓鞘大鼠脑白质保护作用的可能机制。方法将18只SD大鼠随机分为模型组、治疗组、对照组,每组各6只,给予模型组及治疗组大鼠侧脑室注射EB制备脱髓鞘模型,对照组以相同方法注射等体积生理盐水。造模后,治疗组每天经腹腔注射NBP注射液,模型组及对照组腹腔注射等体积生理盐水,10 d后取脑组织制作石蜡切片,行HE染色、LFB髓鞘染色观察大鼠侧脑室周围胼胝体脱髓鞘情况,免疫组化法检测侧脑室周围胼胝体Bcl-2、Bax表达的情况。结果模型组HE染色示胼胝体结构不清、完整性破坏、大量空泡形成,LFB染色示大片区域髓鞘未被蓝染(提示脱髓鞘),与模型组相比,治疗组胼胝体空泡形成较少,髓鞘脱失率明显降低(17.36%vs. 42.05%,P0.05);免疫组化结果显示,治疗组胼胝体Bcl-2阳性表达率较模型组明显增高(69.47%vs. 29.95%,P0.05),Bax阳性细胞表达率明显降低(33.52%vs. 68.61%,P0.05)。结论 NBP可对脱髓鞘大鼠脑白质产生保护作用,减少髓鞘的脱失,其机制可能与上调脑Bcl-2、下调Bax的表达有关。     

利福平对鱼藤酮诱导大鼠的抗多巴胺神经元凋亡作用

目的探讨利福平抗鱼藤酮诱导帕金森病大鼠模型多巴胺神经元凋亡的作用。方法给SD大鼠背部皮下注射鱼藤酮1.5 mg/(kg.d)3周使其黑质多巴胺神经元发生凋亡,同时经灌胃给予利福平30 mg/(kg.d)干预,并通过对大鼠中脑切片进行TUNEL及Bax、Bcl-2和Caspase-3的免疫活性检测以明确利福平抗多巴胺神经元凋亡的作用。结果长期低剂量接触鱼藤酮可诱导SD大鼠中脑黑质部位出现凋亡细胞增加以及Bax、Bcl-2、Caspase-3的免疫活性的改变(P均<0.01),而应用利福平后可明显减轻这些变化(P均<0.01)。结论利福平具有抗鱼藤酮帕金森病大鼠模型的多巴胺神经元凋亡的作用,且此作用是通过上调Bcl-2和Bax的比值、抑制caspase通路而实现的。     

奥扎格雷钠对大鼠局灶性脑缺血血小板内皮细胞黏附分子-1、Bcl-2、Bax表达的影响

目的 观察大鼠局灶性脑缺血后脑组织中血小板内皮细胞黏附分子-1(PECAM-1、CD31)、Bcl-2、Bax表达的改变及奥扎格雷钠对其表达的影响. 方法 采用线栓法制作大鼠大脑中动脉局灶性脑缺血模型,制模成功大鼠随机分为奥扎格雷钠组和生理盐水组,脑组织切片免疫组化染色检测不同时间点PECAM-1、Bcl-2、Bax在各组大鼠脑组织中的表达变化. 结果大鼠大脑中动脉闭塞后脑组织PECAM-1、Bcl-2、Bax的表达明显增高(均P＜0.01).奥扎格雷钠组缺血12h、24h、36h表达PECAM-1较生理盐水组增高,缺血12h、24h 表达Bcl-2较生理盐水组增高,缺血24h、36h表达Bax较生理盐水组增高,均P＜0.01. 结论 脑组织表达的PECAM-1、Bcl-2、Bax分别参与了脑缺血不同时期的病理生理作用;奥扎格雷钠可促进PECAM-1和Bcl-2的表达,抑制Bax的表达.     

右美托咪定调节MAPK/ERK-CREB通路对大鼠海马神经元凋亡的保护作用

目的探讨右美托咪定(DEX)调节MAPK/ERK-CREB通路对大鼠海马神经元凋亡的保护作用。方法通过腹腔注射氯化锂-毛果芸香碱构建癫痫持续状态(SE)大鼠模型,并随机分为4组,每组各10只。SE+DEX组在SE模型构建成功后腹腔注射DEX 1μmol/L,阳性对照组腹腔注射1μmol/L苯巴比妥,药物干预24 h后,通过Nissl法和TUNEL法检测大鼠海马神经元损伤及凋亡情况,Western blot法检测大鼠海马组织中MAPK、p ERK、p CREB蛋白和凋亡相关蛋白caspase-3、Bcl-2、Bax表达。结果与正常对照组相比,SE、阳性对照组、SE+DEX组大鼠Racine分值显著增加(P 0. 05),大鼠海马神经元数、Bcl-2蛋白表达量显著减少(P 0. 05),棕褐色TUNEL阳性细胞数、MAPK、p-ERK、p-CREB、caspase-3、Bax、Bax/Bcl-2蛋白表达量显著增加(P 0. 05)。与SE组相比,阳性对照组、SE+DEX组大鼠Racine分值显著降低(P 0. 05),大鼠海马神经元数、Bcl-2蛋白表达量显著增加(P 0. 05),棕褐色TUNEL阳性细胞、MAPK、p-ERK、p-CREB、caspase-3、Bax、Bax/Bcl-2蛋白表达量显著减少(P 0. 05)。结论 DEX可能通过抑制MAPK/ERK-CREB通路抑制海马神经元凋亡对其有保护作用。     

雌激素对血管性痴呆大鼠认知功能、海马Bcl-2、Bax蛋白表达及神经元凋亡的影响

目的 探讨雌激素对血管性痴呆(VD)大鼠认知功能、海马Bcl-2、Bax蛋白表达和神经元凋亡的影响.方法 60只雄性大鼠随机分为假手术组、VD组和雌激素组,每组20只.采用双侧颈总动脉结扎法制备VD大鼠模型;雌激素组腹腔注射17-β雌二醇(花生油溶解)200 μg/kg,同时假手术组和VD组腹腔注射等量的花生油,均为隔日1次,共30次.60 d后,Y-型迷宫试验检测大鼠学习记忆能力;HE染色观察大鼠海马神经元形态;免疫组化染色检测海马Bcl-2、Bax蛋白的表达,原位缺口末断标记(TUNEL)法检测神经元凋亡程度.结果 与假手术组相比,VD组及雌激素组认知能力明显下降, 海马CA1区Bcl-2、Bax蛋白表达增加,TUNEL 阳性细胞数明显增多(均P<0.001);与VD组比较,雌激素组认知能力改善,海马CA1区Bcl-2蛋白表达明显增加,Bax蛋白的表达明显减少,TUNEL 阳性细胞明显减少 (均P<0.001).结论 17-β雌二醇可调节Bcl-2、Bax蛋白表达而抑制神经元凋亡,有助于改善VD大鼠的认知能力.     

Pregabalin effect on steady-state pharmacokinetics of carbamazepine, lamotrigine, phenobarbital, phenytoin, topiramate, valproate, and tiagabine

By reducing neuronal excitability through selective binding to the α(2)δ subunit of voltage-dependent calcium channels, pregabalin effectively treats epilepsy, chronic pain, and anxiety disorders. To evaluate if pregabalin coadministration affects pharmacokinetics of other antiepileptic drugs, population pharmacokinetic analyses using NONMEM software were performed on data from three epilepsy trials involving seven antiepileptic drugs with pregabalin as add-on therapy. Results demonstrated that pregabalin did not alter the steady-state plasma concentrations of carbamazepine, lamotrigine, phenobarbital, phenytoin, tiagabine, topiramate, and valproate. Furthermore, the small percent change in the population estimate of antiepileptic drug plasma clearance values (-2% to +7%) suggests that pregabalin coadministration exerted no significant effect on the pharmacokinetics of these antiepileptic drugs, with the possible exception of tiagabine (+34.9%). These findings are in agreement with those of previously published reports. A further clarification study is necessary for tiagabine. In conclusion, it appears that pregabalin can be coadministered with other antiepileptic drugs without concern for significantly altering their pharmacokinetic profiles.     

Consecutive Gas Chromatographic Determination of Phenytoin, Phenobarbital, Primidone, Phenylethylmalondiamide, Carbamazepine, Trimethadione, Dimethadione, Ethosuximide, and Valproate from the Same Serum Specimen

A quantitative gas-liquid chromatographic procedure is described for the consecutive determination of phenytoin, phenobarbital, primidone, phenylethylmalondiamide, carbamazepine, trimethadione, dimethadione, ethosuximide and valproate from a single serum specimen of 1.2 ml. After extraction from serum by two different procedures, the anticonvulsants are chromatographed without further purification on a 3% OV 17 column either with or without derivative formation by means of "on-column" methylation. Multiple internal standards are employed in order to enhance the reproducibility of drug-concentration measurement.     

Alteration of proteins regulating apoptosis, Bcl-2, Bcl-x, Bax, Bak, Bad, ICH-1 and CPP32, in Alzheimer''s disease

Recently, apoptosis has been implicated in the selective neuronal loss of Alzheimer's disease (AD). Apoptosis is regulated by the B cell leukemia-2 gene product (Bcl-2) family (Bcl-2, Bcl-x, Bax, Bak and Bad) and the caspase family (ICH-1 and CPP32), with apoptosis being prevented by Bcl-2 and Bcl-x, and promoted by Bax, Bak, Bad, ICH-1 and CPP32. In the present study, we examined the levels of these proteins in the membranous and cytosolic fractions of temporal cortex in AD and control brain. In the membranous fraction, the levels of Bcl-2, Bcl-x_L, Bcl-x_β, Bak and Bad were increased in AD. In the cytosolic fractions, the level of Bcl-x_β was increased, while Bcl-x_L, Bax, Bak, Bad and ICH-1_L were unchanged. CPP32 was not detected in AD or control brain. These findings demonstrate a differential involvement of cell death-regulatory proteins in AD and suggest that Bak, Bad, Bcl-2 and Bcl-x are upregulated in AD brains.     

中国汉族人群SCA1、2、3、6、7、8、10、12、17亚型和齿状核-红核-苍白球-路易体萎缩亚型频率分布

目的 研究中国汉族人群中脊髓小脑性共济失调(SCAs)不同基因亚型的频率分布.方法 运用聚合酶链反应、变性聚丙烯酰胺凝胶电泳、Southern blot、T载体克隆重组DNA技术结合直接测序等技术对559例临床诊断为SCA的患者(363例常染色体显性遗传先证者,196例散发患者)进行了SCA1、SCA2、SCA3/MJD、SCA6、SCA7、SCA8、SCA10、SCA12、SCA17和齿状核-红核-苍白球-路易体萎缩(DRPLA)致病基因多核苷酸病理重复突变检测分析.结果 在363个常染色体显性遗传的SCA(AD-SCA)家系中,发现有15个SCA1家系(4.13%),26个SCA2家系(7.16%),187个SCA3/MJD家系(51.52%),6个SCA6家系(1.65%),7个SCA7家系(1.93%),1个SCA12家系(0.28%)和1个SCA17家系(0.28%),120个SCA家系未明确基因分型(33.06%);在196例散发SCA患者中,发现有2例SCAI患者(1.02%),3例SCA2患者(1.53%),15例SCA3/MJD患者(7.65%),3例SCA6患者(1.53%),173例SCA患者未明确基因分型(88.27%);未发现SCA8、SCA10和DRPLA型患者.结论 在中国汉族人群中SCA3/MJD为最常见的SCA亚型,其次为SCA2、SCA1、SCA7和SCA6,SCA12和SCA17比较少见,SCA8、SCA10和DRPLA罕见,SCA17亚型为国内首次报道.部分AD-SCA家系存在其他致病基因的作用,大部分散发SCA患者除遗传因素外还存在其他致病因素.     

Mass spectrometric identification of Cu, Zn, Fe, Co, Mn, Mg, and Pb in mammalian brain.

Combining the techniques of thin-layer chromatography (TLC) and mass spectrometry, we unambiguously identified the trace metals Cu, Zn, Fe, Pb, Mn, Co, and Mg in the brain of a female human who had no evidence of any pathologic disease in the central nervous system, and in brains from mouse, rat, guinea pig, and rabbit. These trace metals were also found in anatomic regions of human brain: cortex (gray), cortex (white), caudate nucleus, putamen, hippocampus, and thalamus, and in anatomic regions of rat brain: hypothalamus, cerebellum, stem striatum, and "the rest." The metals were characterized from the color and Rf values of their tetraphenylporphyrin chelates on TLC and from the mass and pattern of molecule ion cluster of the mass spectrum. The unexpected presence of lead in the brain is discussed.     

Prevalence, knowledge, attitude, and practice of epilepsy in Kerala, South India

PURPOSE: To ascertain the prevalence and pattern of epilepsy and to characterize and quantify knowledge, attitude, and practice (KAP) toward epilepsy among the people of the state of Kerala, which is distinguished from the rest of India by a high level of literacy and health awareness of its population. METHODS: We conducted a door-to-door survey covering the entire population of 238,102 people residing in 43,681 households in a semiurban area of central Kerala. The screening questionnaire administered by medical social workers had a sensitivity of 100% for identifying persons with epilepsy. Neurologists examined all the individuals suspected of having epilepsy. We evaluated KAP toward epilepsy among 1,118 subjects (439 males and 679 females; mean age, 33.3 years; age range, 15-85 years) from households without epilepsy in the study area. RESULTS: Through a three-phased survey, we ascertained 1,175 cases (616 males and 559 females) with active epilepsy, providing a crude point prevalence ratio of 4.9 cases per 1,000 people and an age-adjusted prevalence ratio of 4.7 cases per 1,000 population. The highest age-specific prevalence rate of 6.5 per 1,000 occurred in the 10- to 19-year-old age group. Sex-specific prevalence rates did not significantly differ. The proportion of generalized and localization-related epilepsies was 58.8% and 30.6%, respectively. Ninety-nine percent of the KAP respondents had read or heard about epilepsy. Thirty-one percent and 27% thought epilepsy was a hereditary disorder and a form of insanity, respectively. About 40% of the respondents felt that individuals with epilepsy could not be properly educated or employed. Eleven percent would object to their children having contact with epileptic children. CONCLUSIONS: The prevalence and pattern of epilepsy in central Kerala, South India, do not differ from that of developed countries. Although the awareness of epilepsy among the people of Kerala was comparable to that of developed countries, the attitudes were much more negative. The need for educating the people of Kerala on epilepsy and for incorporating an adequate knowledge of epilepsy in the school curricula cannot be overemphasized.     

Plant inhibitors of serine proteinases: Hageman factor fragment, kallikreins, plasmin, thrombin, factor Xa, trypsin, and chymotrypsin

Numerous plants were tested for inhibitors of human Hageman factor fragment (HF_f), plasma kallikrein, urinary kallikrein, plasmin, thrombin, porcine pancreatic kallikrein, and bovine Factor X_a, trypsin, and chymotrypsin. Pumpkin seeds and iris bulbs contain trypsin inhibitors which specifically inhibit HF_f. Flower bulbs—especially those of tulip, lily, hyacinth, and calla—are hitherto unrecognized rich sources of inhibitors with different inhibitory spectrums and physicochemical properties.