FTY720对EAE小鼠脑组织NO含量和iNOS表达影响的研究

目的探讨芬戈莫德(fingolimod,FTY720)对试验性自身免疫性脑脊髓炎(EAE)小鼠脑组织中一氧化氮(NO)含量和诱导型一氧化氮合酶(i NOS)表达的影响。方法 48只雌性C57BL/6小鼠随机平均分为3组,EAE组运用MOG35-55构建EAE小鼠模型;CFA组由生理盐水代替MOG35-55构建模型;FTY720干预组在EAE基础上给予FTY720腹腔注射,CFA组、EAE组给予生理盐水腹腔注射。HE染色和LBF染色观察炎症情况和脱髓鞘情况,ELLISA检测小鼠脑组织中的NO含量,RT-PCR检测的诱导型一氧化氮合酶(i NOS)mRNA表达。结果 FTY720组EAE小鼠较EAE组临床症状减轻,炎症程度和脱髓鞘程度减轻(P<0.05)。FTY720组小鼠脑组织NO含量较EAE组降低(P<0.05),i NOS mRNA表达量降低(P<0.05)。结论 FTY720能抑制EAE小鼠脑组织中i NOS mRNA表达,从而减少NO含量。     

芬戈莫德对实验性自身免疫性脑脊髓炎小鼠脑组织基质金属蛋白酶-9表达影响的研究

目的探讨芬戈莫德对实验性自身免疫性脑脊髓炎(EAE)小鼠脑组织基质金属蛋白酶-9(MMP-9)mRNA及蛋白表达的影响。方法 48只雌性C57BL/6小鼠随机分为完全弗氏佐剂(CFA)组、EAE组、芬戈莫德组。运用髓鞘少突胶质细胞糖蛋白35-55构建EAE小鼠模型,小鼠神经功能缺损评分达到1分或免疫第14天,芬戈莫德组腹腔注射芬戈莫德10 mg·kg-1,相应的CFA组、EAE组给予生理盐水。观察小鼠行为学变化,中枢炎症和脱髓鞘情况,脑组织中的MMP-9 mRNA及蛋白表达。结果芬戈莫德组小鼠临床症状减轻,体重下降减少,潜伏期延长,发病率降低,炎性病灶数减少,脱髓鞘程度减轻。芬戈莫德组MMP-9 m RNA和蛋白表达水平较EAE组降低。结论芬戈莫德能抑制EAE小鼠脑组织中MMP-9 mRNA及蛋白表达水平。     

2-BFI对EAE小鼠iNOS和COX-2 mRNA表达的影响

目的探讨咪唑啉2受体(I2R)高选择性高亲和力配体2-BFI对实验性自身免疫性脑脊髓炎(EAE)小鼠诱导型一氧化氮合酶(iNOS)和环氧化酶-2(COX-2)mRNA表达的影响。方法使用髓鞘少突胶质细胞糖蛋白(MOG35-55)抗原诱导EAE小鼠模型。采用临床症状评分、病理学检查和反转录-聚合酶链反应(RT-PCR)观察完全福氏佐剂(CFA)对照组、EAE组和2-BFI干预组小鼠行为学、中枢炎性细胞浸润及iNOS和COX-2mRNA表达变化。结果 2-BFI干预组较EAE组临床症状明显减轻(P<0.01),中枢炎性细胞浸润显著减少(P<0.01),iNOS和COX-2的mRNA表达水平降低(P<0.05,P<0.01)。结论 I2R高选择性配体2-BFI对EAE小鼠具有一定保护作用,其作用机制可能与降低iNOS和COX-2 mRNA表达有关。     

基质金属蛋白酶9在实验性自身免疫性脑脊髓炎大鼠发病过程中的动态表达

目的通过检测基质金属蛋白酶-9(MMP-9)在EAE大鼠发病过程中不同阶段外周血和中枢神经系统中的表达水平及动态变化,以探讨其在多发性硬化发病过程中的作用及机制。方法 Wistar雌性大鼠80只,分为模型组(EAE组)、完全福氏佐剂组(CFA组),分别于免疫后6天、8天、10天、12天、14天、16天、18天及20天取视交叉处脑组织和脊髓腰膨大段行HE染色观察炎性细胞的浸润状况;免疫组化法检测脑和脊髓组织中MMP-9的表达,酶联免疫吸附实验测定血清中MMP-9的含量。结果 EAE组大鼠脊髓和脑组织中MMP-9阳性细胞数及血清中MMP-9水平均显著高于同期CFA组(P<0.05);EAE 10天组与EAE 6天、14天、18天及20天组比较,大鼠脊髓腰膨大处MMP-9阳性细胞数较多(P<0.05);EAE 12天组与EAE 6天组、8天组、14天组、16天组、18天组及20天组比较,大鼠脑组织视交叉处MMP-9阳性细胞数较多(P<0.05);EAE 12天组与EAE 6天组、8天组、10天组、14天组、16天组、18天组及20天组相比较,大鼠血清中MMP-9含量较高(P<0.05)。结论在EAE大鼠发病前期即有中枢组织内MMP-9的高表达,且EAE大鼠脊髓中MMP-9的高表达要早于脑组织和外周血,但MMP-9表达的高峰在第12天与病理变化和疾病进展是同步的。     

新型Rho激酶抑制剂FSD-C11化合物抑制EAE的作用研究

目的探索新型Rho激酶抑制剂FSD-C11化合物治疗实验性自身免疫性脑脊髓炎(EAE)的有效性及可能的作用机制,为今后可能的临床治疗提供实验依据。方法采用小鼠髓鞘少突胶质细胞糖蛋白35-55(MOG35-55)多肽诱导雌性C57BL/6小鼠建立EAE模型,于免疫后第3天起FSD-C11组按体质量40mg/(kg·d)腹腔注射FSD-C11化合物,EAE组注射等量生理盐水,实验期间每天定时记录两组小鼠临床症状评分及体质量变化。免疫后第28天取小鼠脊髓进行HE和髓鞘染色,流式细胞术检测脾细胞M1和M2型巨噬细胞表型,Western blot检测脑组织中诱导型一氧化氮合酶(iNOS)和磷酸化核蛋白因子κB(p-NF-κB)的表达。结果FSD-C11化合物可延迟小鼠的起病时间,降低发病率,减轻临床症状,减少体质量丢失;与EAE组小鼠相比,FSD-C11可减少脊髓炎性细胞浸润和髓鞘脱失(P0.05);抑制致炎性的M1型巨噬细胞,增加抗炎性和保护性的M2型巨噬细胞;抑制脑组织中iNOS和p-NF-κB蛋白的表达。结论新型Rho激酶抑制剂FSD-C11化合物在治疗EAE中显现出很好的潜力,其作用机制可能与调节巨噬细胞极性、抑制炎性反应有关。     

2-(2-苯并呋喃基)-2-咪唑啉对实验性自身免疫性脑脊髓炎小鼠小胶质细胞活化及氧化应激的影响

目的观察2-(2-苯并呋喃基)-2-咪唑啉(2-BFI)对实验性自身免疫性脑脊髓炎(EAE)小鼠CNS小胶质细胞活化及氧化应激的影响。方法将27只C57BL/6小鼠随机分为正常对照组、EAE组和2-BFI干预组,每组9只。采用MOG_(35-55)免疫法制作经典EAE模型。2-BFI干预组小鼠于EAE造模后当日起腹腔注射2-BFI 20 mg/kg,2次/d,连续14 d。正常对照组和EAE组以等量生理盐水代替。每日观察并记录动物的行为学变化、进行神经功能缺损评分。免疫后第20 d处死各组小鼠,采用HE染色和LFB染色观察组织学变化;免疫组化法测定诱导型小鼠CNS中一氧化氮合成酶(iNOS)蛋白和离子钙接头分子(Iba-1)的表达,比色法检测丙二醛(MDA)的含量;并对活化的小胶质细胞数量和iNOS蛋白表达水平进行线性相关分析。结果正常对照组小鼠未见神经系统受损的表现。与EAE组相比,2-BFI干预组日均神经功能缺损评分明显降低,CNS脊髓炎性细胞浸润明显减少,髓鞘脱失严重程度明显减轻,活化的小胶质细胞数量减少,iNOS蛋白表达明显减少,MDA含量降低(P0.05~0.01)。相关分析结果显示,iNOS表达水平与活化的小胶质细胞数量呈正相关(r=0.596,P0.01)。结论 2-BFI能减轻EAE小鼠临床症状和组织学改变,2-BFI对EAE小鼠的神经保护作用于小胶质细胞激活和减轻氧化应激相关。     

二甲双胍通过调节Treg细胞反应治疗实验性自身免疫性脑脊髓炎

目的给予实验性自身免疫性脑脊髓炎(EAE)小鼠应用二甲双胍(MET)干预性治疗。观察MET对EAE小鼠发病情况及体内Treg细胞反应的作用。方法将雌性C57BL/6小鼠随机分为正常对照组、EAE模型组和MET治疗组,采用髓鞘少突胶质细胞糖蛋白MOG35-55免疫小鼠建立EAE模型。自身免疫后第1天开始,按100mg·kg~(-1)·d~(-1)给予MET治疗组腹腔注射。正常对照组及EAE模型组小鼠给予等量的生理盐水每日腹腔注射作为对照。应用Knoz评分观察小鼠的神经功能评分,流式细胞学检测方法检测小鼠脾细胞中Treg细胞比例,ELISA方法检测脾细胞培养上清及血清中IL-10、TGF-β含量。qPCR方法检测小鼠脾及脊髓中Treg细胞转录因子Foxp3 mRNA表达水平。结果与EAE模型组相比,MET治疗组发病程度减轻(P0.01);脾细胞中Treg细胞比例增高(P0.01),脾细胞培养上清及血清中IL-10、TGF-β含量增加(P0.01);脾组织中Foxp3mRNA表达水平升高(P0.01);脊髓组织中Foxp3mRNA表达水平升高(P0.01)。结论 MET通过提高外周免疫器官及中枢神经系统的Treg细胞数量,增加抑制炎症因子的表达而达到对EAE模型小鼠的保护作用。     

雷公藤内酯醇对实验性自身免疫性脑脊髓炎大鼠行为和MMP-9表达的影响

目的:观察雷公藤内酯醇(Tri)对实验性自身免疫性脑脊髓炎(EAE)大鼠中枢神经系统(CNS)中MMP-9表达的影响。方法:建立雌性EAE大鼠模型,将大鼠随机分为4组:治疗组(EAE Tri组);阳性对照组(EAE NS组);正常对照组(CFA Tri组、CFA NS组),观察Tri对EAE大鼠发病情况的影响,并通过免疫组化法观察Tri对EAE大鼠CNS中MMP-9表达的影响。结果:与EAE NS组比较,EAE Tri组的发病率明显降低,临床评分的平均值和最高值均降低,大鼠脑脊髓白质内炎性细胞浸润和病灶数比EAE NS组明显减少。免疫组化:EAE Tri组大鼠CNS中的MMP-9表达比EAE NS组明显减低。正常对照组大鼠均未发病,CNS中也无炎性细胞浸润及MMP-9表达。结论:Tri能抑制EAE大鼠CNS中MMP-9的表达,减轻EAE大鼠临床症状、降低发病率。     

选择性一氧化氮合成酶抑制剂对脑组织保护作用研究

目的：探讨创伤性脑损伤后脑组织中一氧化氮（NO）含量变化与脑组织病理及超微结构变化之间的关系，及选择性一氧化氮合成酶抑制剂（iNOS）1400W(N-[3-(aminomethyl)benzyl]acetamidine)对脑组织的保护作用。方法：114只大鼠随机分成正常组、假手术组、生理盐水治疗组和1400W治疗组，建立大鼠自由落体脑外伤模型，伤后18h开始分别给予生理盐水和1400W腹腔内注射，每8h一次。测定各组不同时期脑组织NO含量，同时观察脑组织病理及超微结构的改变，将结果进行比较。结果：生理盐水治疗组NO含量于伤后6h开始升高，48h达到高峰，以后逐渐下降。1400W治疗组NO含量降低，与生理盐水治疗组结果比较有统计学意义，72h后逐步恢复正常。生理盐水治疗组组织病理学检查和超微结构检查结果与NO改变同步，1400W治疗组较生理盐水治疗组明显改善。结论：NO对脑外伤后继发性损伤起着重要作用，1400W对脑外伤后脑组织有显的保护作用。     

载脂蛋白E拟肽对实验性变态反应性脑脊髓炎小鼠基质金属蛋白酶-9和基质金属蛋白酶组织抑制因子-1表达的影响

目的探讨载脂蛋白E(Apo E)拟肽对实验性变态反应性脑脊髓炎(EAE)小鼠基质金属蛋白酶-9(MMP-9)和基质金属蛋白酶组织抑制因子-1(TIMP-1)表达的影响。方法将30只雌性C57BL/6J小鼠随机分为Apo E拟肽组、EAE组和正常组,每组10只小鼠。EAE模型通过以髓鞘少突胶质细胞糖蛋白多肽35-55为抗原诱导。Apo E拟肽组在免疫后第2 d到30 d每隔2 d按5 mg/(kg·d)背部皮下注射Apo E拟肽。EAE组和正常组均以等体积生理盐水替代。免疫后第0~35 d每日对小鼠进行神经功能评分。免疫后第35d解剖小鼠,分离大脑和脊髓并行HE染色。采用免疫组化染色法检测各组小鼠大脑、脑干和脊髓的MMP-9和TIMP-1的表达。结果正常组小鼠均未发病。Apo E拟肽组、EAE组的小鼠全部发病,但各有1只小鼠发病后死亡。Apo E拟肽组与EAE组的发病潜伏期差异无统计学意义(P=0.72)。Apo E拟肽组的神经功能评分在峰值和慢性期(第35 d)均明显低于EAE组(均P0.05)。HE染色示,正常组未见炎症细胞浸润;EAE组小鼠大脑、脑干和脊髓均有不同程度的炎性细胞浸润,以脑干和脊髓较为明显;Apo E拟肽组小鼠CNS炎性细胞浸润相对于EAE组明显减少。EAE组小鼠大脑、脑干和脊髓的MMP-9表达均高于正常组(均P0.05)。Apo E拟肽组小鼠大脑和脊髓的MMP-9表达要明显低于EAE组(均P0.05),其中Apo E拟肽组小鼠中脑和脊髓的MMP-9表达与正常组相比无明显差异(均P0.05)。正常组小鼠脊髓TIMP-1的表达明显高于EAE组和Apo E拟肽组(均P0.05)。而Apo E拟肽组与EAE组小鼠大脑、脑干和脊髓TIMP-1表达的差异均无统计学意义(均P0.05)。结论 Apo E拟肽能通过抑制大脑和脊髓MMP-9的表达改善EAE小鼠的症状。     

Denervation study of synapses of organ of corti of old world monkeys

Fine structural characteristics of synapses in the spiral organ of Corti were examined, with reference to differences between inner and outer haircell systems, and to location of neurons of origin of efferent axons. Surgical interruption of crossed olivocochlear bundle, of vestibular nerve, of facial nerve, and excision of superior cervical ganglia were used to determine the pathways of efferent axons. Interruption of the vestibular nerve near the brainstem results in degeneration of all efferent terminals on outer hair cells. Mid-line lesions at, and caudal to, the facial colliculus result in degeneration of about half of these efferent terminals. Efferent synaptic bulbs to the inner hair-cell system are small, of the order of one micron, and form type 2 junctions with afferent dendrites. They tend to have more large dense-core vesicles (about 80 nm) than the large efferent terminals of the outer hair-cell system, and appear to be the terminals of axons in the habenula perforata, which exhibit varicosities laden with large dense core vesicles. The varicosities are unaffected by excision of the superior cervical ganglia. So far as our material can reveal, it appears that the varicosities in the habenula perforata do not survive vestibular root interruption, nor do the efferent processes in the internal spiral bundle or at the base of inner hair cells. Most interestingly, the afferent processes of the inner hair-cell system, as identified for example by their relation to pre-synaptic bodies in the inner hair cells, are subject to a trans-synaptic reaction after severance of the vestibular root. They undergo a dramatic cytological transformation, characterized by increase of volume, engorgement with microtubules, microfilaments, microvesicles of various sizes, and clusters of lysosomes. Thus, both the efferent and afferent terminals of the inner hair-cell system show marked cytological differences from the corresponding terminals of the outer hair cell system.     

Mechanism of action of tubocurarine on nicotinic cholinoreceptors of neurons of the sympathetic ganglia of rats

Tubocurarine (Tc) effect on membrane currents elicited by acetylcholine (ACh) was studied in isolated superior cervical ganglion neurons of rat using patch-clamp method in the whole-cell recording mode. The "use-dependent" block of ACh current by Tc was revealed in the experiments with ACh applications, indicating that Tc blocked the channels opened by ACh. Mean lifetime of Tc-open channel complex, tau, was found to be 9.8 +/- 0.5 s (n = 7) at -50 mV and 20-24 degrees C. tau exponentially increased with membrane hyperpolarization (e-fold change in tau corresponded to the membrane potential shift by 61 mV). Inhibition of the ACh-induced current by Tc (3-30 microM/1) was completely abolished by membrane depolarization to the level of 80-100 mV. Inhibition of ACh-induced current was augmented at increased ACh doses. It is concluded that the open channel block produced by Tc is likely to be the only mechanism for Tc action on nicotinic acetylcholine receptors in superior cervical ganglion neurons of rat.     

The effect of age of onset of PD on risk of dementia

Background Dementia occurs in the majority of patients with Parkinson’s disease (PD). Late onset of PD has been reported to be associated with a higher risk for dementia. However, age at onset (AAO) and age at baseline assessment are often correlated. The aim of this study was to explore whether AAO of PD symptoms is a risk factor for dementia independent of the general effect of age. Methods Two community-based studies of PD in New York (n = 281) and Rogaland county, Norway (n = 227) and two population-based groups of healthy elderly from New York (n = 180) and Odense, Denmark (n = 2414) were followed prospectively for 3–4 years and assessed for dementia according to DSM-IIIR. All PD and control cases underwent neurological examination and were followed with neurological and neuropsychological assessments. We used Cox proportional hazards regression based on three different time scales to explore the effect of AAO of PD on risk of dementia, adjusting for age at baseline and other demographic and clinical variables. Findings In both PD groups and in the pooled analyses, there was a significant effect of age at baseline assessment on the time to develop dementia, but there was no effect of AAO independent of age itself. Consistent with these results, there was no increased relative effect of age on the time to develop dementia in PD cases compared with controls. Interpretation This study shows that it is the general effect of age, rather than AAO that is associated with incident dementia in subjects with PD. Received in revised form: 22 December 2005     

Limits of deinstitutionalization--perspectives of relatives of psychiatric patients

After a hopeful beginning, the social process of the reintegration of those with severe mental illness has come to a standstill. I am led to wonder whether "the community" really wants to live together with people suffering from severe mental illness, and if so, how closely? As long as the medical treatment of mental illness provided by the general practitioners is fundamentally deficient, as they are not able to prescribe the necessary interventions--such as out-patient psychiatric nursing, and service providers in the out-patient sector are content with offering increasingly intensive forms of care for the less seriously ill at the cost of the Social Welfare System--the reintegration of those with serious mental illness remains an illusion--which is mainly to the benefit of providers of residential care in homes and hostels.