实验性脑出血后脑组织核因子-κB表达和含水量的变化及其相关性

目的 研究实验性脑出血(ICH)后血肿周围脑组织核因子-κB(NF-κB)表达和脑组织含水量的改变及其相关性.方法 采用自体不凝血注人大鼠尾状核制备ICH模型;用免疫组化法检测血肿周围脑组织NF-κB表达;用干湿重法测脑组织含水量.结果 与对照组相比,ICH 6h组血肿周围脑组织NF-κB表达开始明显增加,ICH 48h组达高峰,并持续到ICH 1周(P＜0.01～0.05);脑组织含水量ICH 12h组开始增多,24h组显著增高,72h组达高峰;NF-κB表达和脑组织含水量ICH 24h～1周组与假手术组之间差异有统计学意义(均P＜0.01);ICH后NF-κB阳性细胞数与脑组织含水量呈正相关(r=0.644,P＜0.01),NF-κB表达从开始到高峰均早于脑组织含水量的变化.结论 ICH后脑组织NF-κB表达增加,NF-κB可能通过炎性机制参与了ICH后继发脑水肿的形成.     

实验性脑出血血肿周围组织中Rho激酶的研究

目的观察实验性脑出血(ICH)血肿周围组织中Rho激酶的表达及其与神经功能缺损的关系。方法将ICH模型大鼠随机分为ICH后1d、3d、5d及7d组;在相应时间点处死大鼠前进行神经功能评分;运用免疫组化法和原位杂交法观察每组大鼠血肿周围脑组织中Rho激酶蛋白及mRNA的表达。结果ICH后1d、3d、5d及7d组神经功能评分分别为1.63、2.25、3.13及2.88;血肿周围脑组织Rho激酶蛋白阳性细胞数分别为26.63、30.75、38.88及35.00;血肿周围Rho激酶mRNA阳性细胞数分别为27.25、32.25、40.88及38.00。1d、3d、5d组间神经功能评分、Rho激酶蛋白及mRNA的表达差异有统计学意义(P<0.05～0.01),5d组与7d组间差异无统计学意义;神经功能评分与Rho激酶蛋白及mRNA的表达呈正相关(r1=0.534,r2=0.675,均P<0.01)。结论ICH后血肿周围脑组织中Rho激酶可能被激活,并参与了ICH后血肿周围脑组织的损害过程。     

实验性脑出血血肿周围脑组织基质金属蛋白酶组织抑制剂-1的表达及其与脑水肿的关系

目的探讨实验性脑出血(ICH)血肿周围脑组织基质金属蛋白酶组织抑制剂-1(TIMP-1)的表达及其与脑水肿的关系。方法采用自体血注入法建立大鼠ICH模型,制模后1h、3h、6h、12h、24h、36h、48h、72h、7d共9个时间点分别用干湿重法测血肿周围脑组织含水量,伊文思蓝(EB)检测血-脑屏障(BBB)通透性,Western Blot方法检测血肿周围组织的TIMP-1的表达;分析三者间的关系,并与假手术组相比较。结果与假手术组比较,ICH后各时间点血肿周围脑组织含水量和EB含量均明显增加(P<0.05～0.01),均于ICH48h达到高峰,且二者呈正相关(r=0.940,P<0.01);TIMP-1在ICH12～36h表达明显减低,ICH1～24h TIMP-1表达量与脑组织含水量以及EB含量呈负相关(r=-0.922,-0.950,均P<0.05)。结论大鼠ICH后血肿周围脑组织TIMP-1表达下调,可能促进了ICH早期的脑水肿形成。     

大鼠脑出血后基质金属蛋白酶的动态表达及七叶皂苷钠对其表达的影响

目的 观察大鼠脑出血(ICH)后基质金属蛋白酶-2( MMP-2)、MMP-9蛋白动态表达及其与血肿周围脑组织含水量的关系,以及观察七叶皂苷钠对MMP-2、MMP-9表达的影响.方法 Wistar大鼠250只随机分为4组:正常对照组10只、假手术组、ICH组和七叶皂苷钠治疗组各80只,于制模后6 h、12 h、24 h、48 h、72 h、120 h、7 d、15 d 8个时间点,测定各组血肿周围脑组织含水量、MMP-2和MMP-9的蛋白表达.结果 治疗组神经功能缺损较ICH组明显改善,尤以6 h～7 d时明显(P<0.01～0.05);6～120 h各治疗组血肿周围脑组织含水量较ICH组明显减少(P<0.01～0.05);ICH后MMP-2蛋白表达在6 h达到高峰,12 h时下降,与正常对照组相比具有统计学意义 (均P<0.01),治疗组MMP-2蛋白表达在各时间点较ICH组明显减少(均P<0.01);ICH后MMP-9蛋白表达在6 h开始上升,24～48 h达高峰,72 h时下降,与正常对照组相比,具有统计学意义(均P<0.01),其表达水平与血肿周围脑组织含水量呈正相关 (r=0.949, P<0.05);治疗组MMP-9蛋白表达在各时间点较ICH组明显减少(均P<0.01).结论 大鼠ICH后MMP-2、MMP-9蛋白的表达是ICH后早、中期脑水肿形成主要因素,七叶皂苷钠能降低MMP-2、MMP-9蛋白表达和脑组织含水量,对ICH具有保护作用.     

实验性脑出血后脑组织核因子-κB表达和含水量的变化及其相关性

目的研究实验性脑出血（ICH）后血肿周围脑组织核因子-кB（NF-кB）表达和脑组织含水量的改变及其相关性。方法采用自体不凝血注入大鼠尾状核制备ICH模型;用免疫组化法检测血肿周围脑组织NF-кB表达;用干湿重法测脑组织含水量。结果与对照组相比,ICH6h组血肿周围脑组织NF-кB表达开始明显增加,ICH48h组达高峰,并持续到ICH1周（P〈0.01~0.05）;脑组织含水量ICH12h组开始增多,24h组显著增高,72h组达高峰;NF-кB表达和脑组织含水量ICH24h~1周组与假手术组之间差异有统计学意义（均P〈0.01）;ICH后NF-кB阳性细胞数与脑组织含水量呈正相关（r=0.644,P〈0.01）,NF-кB表达从开始到高峰均早于脑组织含水量的变化。结论ICH后脑组织NF-кB表达增加,NF-кB可能通过炎性机制参与了ICH后继发脑水肿的形成。     

高血压脑出血患者血肿周围血-脑屏障葡萄糖转运蛋白1表达的研究

目的观察高血压脑出血(ICH)患者血肿周围血-脑屏障葡萄糖转运蛋白1(GLUT1)表达的变化。方法采用免疫组化方法检测28例高血压ICH患者不同病程血肿周围脑组织GLUT1的表达情况。结果与对照组相比,ICH组发病12 h GLUT1表达开始增强,24 h、48 h表达最强(均P<0.01);72 h表达明显下降(P<0.01)。结论ICH早期血肿周围GLUT1表达增多,有助于恢复脑的能量代谢。     

脑出血大鼠血肿周围白介素-10的表达及其与脑水肿的相关性

目的 探讨脑出血(ICH)大鼠血肿周围白介素-10(IL-10)的表达及其与脑水肿的相关性.方法 130只SD大鼠随机分为正常对照组、假手术组和ICH组;应用肝素化Ⅶ型胶原酶建立大鼠ICH模型;制模后12 h、24 h、48 h、72 h、7 d及14 d应用Bederson量表评定神经功能缺损程度,干/湿重法测定脑组织含水量,免疫组化方法检测脑组织血肿周围IL-10的表达.结果 (1)ICH组大鼠神经功能缺损评分与脑组织含水量均在ICH 12 h升高,48 h达高峰,7 d后恢复正常.(2)ICH组脑组织IL-10表达ICH 12 h明显升高,14 d达高峰,各时间点间均显著高于正常对照组和假手术组(P＜0.05～0.01).(3)ICH组神经功能缺损评分与脑组织含水量呈正相关(r=0.761,P＜0.01),脑组织IL-10表达与脑组织含水量、神经功能缺损评分呈负相关(r=-0.65,-0.753,均P＜0.01).结论 ICH大鼠血肿周围脑组织IL-10表达逐渐升高,可能参与了ICH后减轻脑水肿、促进神经功能恢复的脑保护作用.     

COX-2在大鼠脑出血损伤机制中的实验研究

目的观察大鼠脑出血后环氧合酶2(COX-2)蛋白动态表达情况与血肿周围脑组织含水量的相互关系,及COX-2抑制剂对ICH大鼠脑水肿和神经功能的影响,探讨COX-2在ICH损伤机制中的作用。方法应用立体定向技术将自体血注入大鼠壳尾核制备脑出血模型,160雄性SD大鼠分为正常对照组、假手术组、脑出血组和尼美舒利治疗组。后3组每组50只,分6h、12h、24h、72h、7d五个时间点进行神经功能障碍评分,测定血肿周边脑组织含水量,免疫组织化学染色观察COX-2蛋白动态表达。结果脑出血组COX-2表达高于正常对照组(P<0.05),治疗组COX-2表达明显低于ICH组(P<0.05)。治疗组脑组织含水量明显低于ICH组(P<0.05)。治疗组各时间点神经功能评分明显高于ICH组(P<0.05)。ICH组COX-2蛋白表达与血肿周围脑组织含水量相关(r=0.911,P<0.001)。结论ICH后COX-2蛋白表达升高促进脑水肿形成,加重神经功能缺失。尼美舒利能抑制COX-2蛋白表达,减轻脑水肿,改善ICH后大鼠神经功能。     

补体在脑出血后脑组织损伤机制中的作用

目的研究补体C9在大鼠实验性脑出血(ICH)后血肿周围组织中的表达情况,探讨补体C9在ICH后脑水肿中的作用以及应用眼镜蛇毒因子(CVF)干预后对血肿周围组织C9表达及脑组织含水量变化的影响。方法采用立体定向技术,将自体不凝血注入大鼠尾状核制备ICH模型,将动物分为假手术组、出血组和CVF干预组,分别在不同时间断头取脑,连续切片分别作补体C9免疫组化染色和HE染色,并进行脑组织含水量测定(干湿重法)。结果ICH后2h血肿周围脑组织开始表达C9,24h达高峰。血肿周围脑组织含水量在ICH后2h开始增加(P<0.05),6h明显增加,24～72h达高峰(P<0.01),此后逐渐回落,1周基本恢复正常水平,脑组织含水量与C9的表达呈正相关关系(r=0.938,P<0.01);对侧半球相应部位及假手术对照组脑组织含水量没有明显变化;经CVF干预后,血肿周围组织C9表达明显下降,干预组与出血组之间比较有显著差异(P<0.01)。CVF干预组脑组织含水量明显低于常规ICH组(P<0.01)。结论脑出血后补体级联激活C9表达明显增加,并证明通过CVF干预后,C9表达下降,脑水肿减轻,能达到神经保护作用。     

大鼠脑出血后脑组织转铁蛋白表达的实验研究

目的　探讨大鼠自发性脑出血 (ICH)后不同时间脑组织中转铁蛋白 (Tf)的表达及其规律。方法　采用立体定向技术注入自体不凝血建立实验性大鼠ICH模型 ,在不同时间断头取脑 ,免疫组化染色检测脑组织中Tf阳性细胞的表达并与对照组比较。结果　ICH后 6h血肿周围及同侧大脑皮质Tf阳性细胞的表达增高 ,72h达高峰 ,然后下降 ,但第 7d时仍高于对照组 (P <0 0 5 )。结论　ICH后血肿周围和同侧大脑皮质Tf阳性细胞表达增高 ,推测Tf可能参与了ICH后神经元的保护并减轻迟发性脑水肿     

视频脑电图在小儿癫痫诊断中的应用

目的评价视频脑电图(video-EEG)在小儿癫诊断中的应用价值。方法对126例具有发作性症状的患儿进行连续8h的包括清醒、睡眠、诱发试验及必要的认知测验的视频脑电图监测。结果经发作期视频脑电图证实,39例初诊为癫性发作的患儿中14例(35%)为非癫性发作;15例其他症状发作中13例(86%)为非癫性发作。64例样放电患儿中51例(80%)确定发作类型,22例(34%)确定癫类型。视频脑电图可发现短暂轻微的癫发作及样放电引起的一过性认知损伤。结论视频脑电图在排除非癫性发作、确定癫性发作的类型、评价脑电-临床关系方面可提供准确可靠的证据,进一步提高癫的临床诊断水平。     

Neuroprotective properties of memantine in different in vitro and in vivo models of excitotoxicity

The pathogenesis of stroke, trauma and chronic degenerative diseases, such as Alzheimer's disease (AD), has been linked to excitotoxic processes due to inappropriate stimulation of the N-methyl-D-aspartate receptor (NMDA-R). Attempts to use potent competitive NMDA-R antagonists as neuroprotectants have shown serious side-effects in patients. As an alternative approach, we were interested in the anti-excitotoxic properties of memantine, a well-tolerated low affinity uncompetitive NMDA-R antagonist presently used as an anti-dementia agent. We explored in a series of models of increasing complexity, whether this voltage-dependent channel blocker had neuroprotective properties at clinically relevant concentrations. As expected, memantine protected neurons in organotypic hippocampal slices or dissociated cultures from direct NMDA-induced excitotoxicity. However, low concentrations of memantine were also effective in neuronal (cortical neurons and cerebellar granule cells) stress models dependent on endogenous glutamate stimulation and mitochondrial stress, i.e. exposure to hypoxia, the mitochondrial toxin 1-methyl-4-phenylpyridinium (MPP+) or a nitric oxide (NO) donor. Furthermore, memantine reduced lethality and brain damage in vivo in a model of neonatal hypoxia-ischemia (HI). Finally, we investigated functional rescue (neuronal capacity to migrate along radial glia) by memantine in cerebellar microexplant cultures exposed to the indirect excitotoxin 3-nitropropionic acid (3-NP). Potent NMDA-R antagonists, such as (+)MK-801, are known to block neuronal migration in microexplant cultures. Interestingly, memantine significantly restored the number of neurons able to migrate out of the stressed microexplants. These findings suggest that inhibition of the NMDA-R by memantine is sufficient to block excitotoxicity, while still allowing some degree of signalling.     

Storage of lipofuscin in neurons in mucopolysaccharidosis

Summary A histochemical and ultrastructural study was made on the brain of a 23-year-old man with Sanfilippo's syndrome. In accordance with previous reports the cortical nerve cells contained a PAS-positive lipid storage substance. This showed intense autofluorescence in UV-light and was positive with various stains for lipofuscin. The storage material appeared ultrastructurally as inclusion bodies composed of short lamellated membranes, granular material, and vacuoles. In addition, concentrically and transversely lamellated membranous cytoplasmic bodies were observed in the nerve cells. It is concluded that the PAS-positive lipid storage material in the neurons was composed partly of lipofuscin in addition to other lipids presumably glycosphingolipids.Supported by a grant from the Expressen Prenatal Research Foundation     

脑电图预测痫性发作研究进展

癫痫(epilepsy)是由脑部神经元高度同步化异常放电所致的临床综合征,系神经系统的常见病,困扰着全世界约1%的人群.每次神经元的阵发性放电或短暂的脑功能异常称为痫性发作(seizures).     

Midazolam in treatment of various types of seizures in children

Midazolam is a recently developed water-soluble benzodiazepine that shares anxiolytic, muscle relaxant, hypnotic and anticonvulsant actions with other members of this class. There are limited studies that midazolam can be used successfully to treat seizures in adults and children. In this study, 0.2 mg/kg intramuscular (IM) midazolam was administered to 11 children (eight boys and three girls), aged 3 days to 4 years (mean age 1.8±1.4 years), with seizures of various types. In all but one child, seizures stopped in 15 s–5 min after injection. No side effects were observed. These results suggest that IM administration of midazolam may be useful in a variety of seizures during childhood, especially in case of intravenous (IV) line problem.     

Role of vincristine in the inhibition of angiogenesis in glioblastoma

Objective: Vincristine, a microtubule-destabilizing drug, was found to exhibit anti-angiogenic effects and anti-tumoral activity. However, the precise mechanism by which vincristine inhibits angiogenesis in glioblastomas is not well understood. Our aim was to investigate whether vincristine affects vascular endothelial growth factor (VEGF) expression in glioblastoma cells and determine whether it is mediated by the downregulation of hypoxia-inducible factor-1α (HIF-1α).

Methods: We investigated the expression of HIF-1α in glioblastoma tissues resected from patients and in human glioblastoma cell lines using immunohistochemistry, Western blot analysis, and immunocytochemistry. In addition to an MTT assay assessing the effect of vincristine on cell proliferation and viability, the effects of vincristine on VEGF mRNA expression and HIF-1α protein were examined using real-time RT-PCR and Western blot analysis under 1% O₂ (hypoxia).

Results: HIF-1α was expressed in the majority of glioblastoma tissues and was detected mainly in the nucleus. Strong immunoreactivity for HIF- 1 α was found often in the hypercellular zones. Under hypoxic conditions, HIF-1α protein levels in the glioblastoma cell lines increased, primarily localizing into the nucleus similar to glioblastoma tissues. Exposure of glioblastoma cells to vincristine resulted in enrichment of the G₂-M fraction of the cell cycle, which suggests that vincristine-mediated growth inhibition of glioblastoma is correlated with mitotic inhibition. Using doses lower than those found to reduce the viability and proliferation of cells by 50% (IC₅₀), vincristine decreased both the expression of VEGF mRNA and the level of HIF-1α protein in hypoxic glioblastoma cells. In addition, following exposure to vincristine, the expression of VEGF mRNA was correlated with HIF-1α protein levels.

Conclusions: Our results suggest that the mechanism by which vincristine elicits an anti-angiogenic effect in glioblastomas under hypoxic conditions might be mediated, in part, by HIF-1α inhibition.     

The Burden of Agoraphobia in Worsening Quality of Life in a Community Survey in Italy

ObjectiveCurrent nosology redefined agoraphobia as an autonomous diagnosis distinct from panic disorder. We investigated the lifetime prevalence of agoraphobia, its association with other mental disorders, and its impact on the health-related quality of life (HR-QoL). MethodsCommunity survey in 2,338 randomly selected adult subjects. Participants were interviewed with the Advanced Neuropsychiatric Tools and Assessment Schedule (ANTAS), administered by clinicians. The diagnoses were based on the ICD-10 criteria. The Short-Form Health Survey (SF-12) was used to quantify HR-QoL. ResultsIn the sample, 35 subjects met the criteria for agoraphobia (1.5%), with greater prevalence among women (2.0%) than men (0.9%): odds ratio (OR) 2.23; 95% CI: 1.0-5–2. Agoraphobia was more often seen among those with (n=26; 1.1%) than without (n=9; 0.4%) panic disorder: OR=8.3; 2.9–24.4. Co-morbidity with other mental disorders was substantial. The mean score of SF-12 in people with agoraphobia was 35.2±7.8, with similar levels of HR-QoL in people with (35.3±7.9) or without (34.8±7.3) panic disorder: ANOVA: F(1;33)=0.0; p=1.00. ConclusionOne out of seventy people may suffer from agoraphobia in their lifetime. The attributable burden in terms of HR-QoL is substantial and comparable to the one observed for chronic mental disorders such as major depression, post-traumatic stress disorder, or obsessive-compulsive disorder.     

Characteristics of nociceptors in the periodontium--an in vitro study in rats

Recent studies have indicated that nociceptors can be classified into various types according to their physiological properties. These studies have clarified that the frequency distribution of various nociceptor types is different among body sites and animal species. In the present study, we investigated the physiological properties of rat's periodontal nociceptors in an in vitro jaw-nerve preparation. Responses were recorded from functional single filaments in the inferior alveolar nerve. To determine the nociceptor type, calibrated von Frey filaments, heat, and bradykinin (BK) stimuli were used. We found five subtypes of nociceptors in the periodontal ligaments of the lower incisor: Adelta-high threshold mechanonociceptors (Adelta-HTM, n=28), Adelta-mechanoheat nociceptors (Adelta-MH, n=6), Adelta-polymodal nociceptors (Adelta-POLY, n=26), C-high threshold mechanonociceptors (C-HTM, n=3) and C-polymodal nociceptors (C-POLY, n=4). Most nociceptors were Adelta-innervated, while only a small number of C-innervated nociceptors were found. The present results suggest that periodontal nociceptors transmit mainly fast pain, and may thus play a role in rapid detection of injure-related stimuli during mastication.     

自噬参与帕金森病发病的研究进展

近年来,蛋白质的降解障碍被认为是帕金森病（Parkinson’Sdisease,PD）发病过程中的重要因素,人们已经公认泛素一蛋白酶体系统（ubiquitin--pro—teasomesystem,UPS）功能异常或衰竭能够导致细胞内异常蛋白蓄积、细胞功能障碍,甚至细胞凋亡。与此同时,蛋白降解的另一条途径——自噬-溶酶体途径（autophagy—lysosomepathway,ALP）也已成为了生命科学领域的研究热点,自噬与神经变性疾病,尤其是PD的关系日益受到人们的重视。