七叶皂苷钠对实验性大鼠脑出血后MMP-2/9表达的影响

目的探讨七叶皂苷钠对实验性脑出血大鼠脑组织中基质金属蛋白酶-2／9(MMP-2／9)表达的影响。方法采用胶原酶复制大鼠脑出血模型,60只SD大鼠随机分为模型组、治疗组A及治疗组B,通过免疫组化和原位杂交技术动态测定血肿周围脑组织中MMP-2／9的mRNA和蛋白表达,同时采用七叶皂苷钠进行治疗。结果模型组各时点出现大量MMP-2／9的蛋白和mRNA阳性细胞,且MMP-9的蛋白和mRNA表达在24～48h达高峰。 MMP-9蛋白与其mRNA表达呈正相关,MMP-2蛋白与其mRNA表达呈正相关。七叶皂苷钠可明显抑制二者的表达(P<0．01),存在一定的量效关系,呈负相关。结论七叶皂苷钠可抑制大鼠脑出血早期诱导的血肿周围脑组织中的MMP-2／9表达上调。     

实验性脑出血后脑水肿的动态变化及其与MMP-9的关系

目的探讨脑出血后脑水肿和MMP-9表达的动态变化,并初步探讨其关系。方法健康雄性Wistar大鼠56只,将动物随机分成假手术组和脑出血组,大鼠尾壳核区注入自体非肝素抗凝动脉血建立脑出血模型,采用干湿重法测量脑组织水含量;免疫组织化学方法观察术后不同时间点MMP-9的动态变化。结果血肿形成后脑水肿产生迅速,从脑出血后3h开始增加,48~72h达高峰;脑出血后12h时,血肿周围组织中开始出现大量的深棕黄色阳性染色细胞,术后72h达高峰,以后逐渐下降,至120h时仍维持在较高水平。结论脑出血后脑水肿可能与MMP-9的活化有关。     

大鼠脑出血后脑组织MMP-2、MMP-9 表达的实验研究

目的　探讨大鼠脑出血后不同时程脑组织中的基质金属蛋白酶 - 2 ( MMP- 2 )、基质金属蛋白酶 - 9( MMP- 9)的表达及其规律。方法　采用立体定向技术制作大鼠脑出血模型 ,在脑出血后不同时间分别断头取脑 ,免疫组化法测定脑组织 MMP- 2、MMP- 9的表达量。结果　 ( 1)与假手术对照组比较 ,脑出血后 6 h血肿周围可见到微血管内皮表达 MMP- 9( P<0 .0 1) ,48h达高峰 ,至 5～ 7d后下降 ,但仍高于假手术对照组 ( P<0 .0 1) ,2周时降至零表达 ;12～ 2 4h中性粒细胞亦表达 MMP- 9;( 2 ) MMP- 2的表达要迟于 MMP- 9,2 4h可见到少量的 MMP- 2表达 ( P>0 .0 5 ) ,5～ 7d时逐渐达高峰 ( P<0 .0 1) ,主要在巨噬细胞上表达 ,2周时仍保持较高水平 ( P<0 .0 1)。结论　脑出血后出血侧血肿边缘有 MMP- 9、MMP- 2表达 ,推测 MMP- 9在急性期参与了脑水肿的形成 ,而 MMP- 2在脑出血后期可能参与了脑组织的修复.     

凝血酶对大鼠脑内MMP-9、MMP-2表达的影响

目的探讨凝血酶对大鼠脑内MMP-9、MMP-2表达的影响。方法Wistar大鼠随机分为假手术组及实验组。实验组脑内注入凝血酶,在不同时间点采用干湿重法测脑水含量,免疫组化方法检测脑内MMP-9、MMP-2的表达。假手术组注入等量生理盐水。结果脑组织水含量在凝血酶注入后6h开始增加,3d达高峰。MMP-9阳性细胞在注射凝血酶后6h即开始表达增加,与对照组相比差异显著(P<0.01),3d达高峰,随后持续下降。阳性微血管数也有相似的变化趋势。MMP-2阳性细胞在注射凝血酶后1d才开始有少量表达,后持续增多,5d达高峰,随后有所下降,但14d仍有明显表达,与对照组相比差异显著(P<0.01)。阳性微血管数也有相似的变化趋势。结论凝血酶在脑出血后脑水肿及脑组织损伤中起了关键的作用,MMP-9、MMP-2参与了急性期脑水肿、炎症反应等过程,MMP-2在损伤修复中可能有重要作用。     

实验性脑出血大鼠脑内Caspase-3mRNA的表达作用研究

目的动态观察实验性脑出血大鼠脑内血肿周围神经细胞凋亡情况和Caspas-3蛋白及mRNA表达水平的变化,探讨脑H{血后血肿周围神经细胞损伤机制。方法健康雄性Wistar大鼠随机分为生理盐水对照组、假手术组和脑出血模型组,分为术后3h,6h,12h,24h,48h,3d,5d,7d共8个时相点,采用尾状核注射自体非抗凝动脉血复制大鼠脑出血模型,术后制作冰冻切片,对切片进行TUNEL染色,以及Caspase-5免疫组化和原位杂交染色,之后利用图像分析仪,观察阳性细胞数。结果脑出血后3h血肿周围尚无凋亡细胞出现,6h有凋亡发生,以后逐渐增多,3d达高峰后逐渐下降,生理盐水对照组仅有少量TUNEL阳性细胞。假手术组及生理盐水对照组3h无Caspase-3蛋白和mRNA表达,生理盐水对照组6h以后有少量表达,脑出血模型组在6hCaspase-3蛋白和mRNA开始表达,3d时Caspase-3的蛋白达到高峰,5d以后逐渐下降,24hCaspase-3mRNA表达达高峰,5d以后逐渐下降。脑出血后血肿周围脑组织Caspase-3蛋白的表达与TUNEL阳性细胞数呈正相关（r=0．515,P〈0．05）;Caspase-3蛋白表达与mRNA表达呈正相关（r=0．625,P〈0．05）。结论脑出血后血肿周围神经细胞损伤有凋亡机制参与,在出血后6h发生凋亡,第3天达高峰。Caspase-3的表达在Caspase-5蛋白水平变化趋势与脑m血后细胞凋亡的趋势一致,Caspase-3的mRNA水平的表达高峰时间先于Caspase-3蛋白的表达及凋亡的发生,提示Caspase-3的表达决定脑出血后细胞凋亡的发生,在脑出血后细胞凋亡中起促进作用。     

脑出血灶周组织ICAM-1与MMP-9表达的实验研究

目的研究大鼠实验性脑出血后血肿周围脑组织细胞间粘附分子-1（ICAM-1），基质金属蛋白酶-9（MMP-9）的表达。方法采用立体定向技术将自体不凝血注入大鼠尾状核区制备脑出血模型，免疫组化染色法检测血肿周围脑组织ICAM-1，MMP-9的表达。结果脑出血后6h血肿旁有少量ICAM-1表达阳性细胞，12h开始增多，3d达高峰；脑出血后6h血肿周围就有较多MMP-9表达阳性细胞，2d时阳性细胞最多；脑出血后ICAM-1与MMP-9的表达呈正相关（y=0．768，P〈0．05）。结论血肿周围组织ICAM-1、MMP-9表达上调提示两者可能参与了脑出血后继发性脑损伤。     

脑出血后血脑屏障紧密连接蛋白occludin的表达变化

目的 探讨脑出血后血脑屏障微血管内皮细胞间紧密连接蛋白occludin的表达变化. 方法 将SD雄性大鼠按随机数字表法分为正常对照组和脑出血组,再按时间因素将脑出血组分为出血后6h、24h、48h、72h、7d、14d6个亚组.采用脑内注入自体血法制作脑出血模型.HE染色观察脑出血后血肿周围脑组织的形态学改变;透射电镜观察脑出血后血肿周围紧密连接的超微结构改变;免疫荧光染色检测脑出血后血肿周围紧密连接蛋白occludin的表达分布状况;定量RT-PCR检测脑出血后血肿周围脑组织中occludin mRNA的表达状况. 结果 与正常对照组相比,脑出血组血肿周围脑组织出现水肿,在48 h左右尤为明显,局部可见明显脑细胞坏死及炎细胞浸润.脑出血后血肿周围紧密连接发生明显破坏,内皮细胞间出现裂隙.免疫荧光染色结果显示:正常对照组紧密连接蛋白occludin呈强阳性表达.脑出血后6hoccludin的表达即开始下降,呈阳性表达;脑出血后24～72h occludin的表达维持在较低水平,呈弱阳性表达.定量RT-PCR结果显示:脑出血后血肿周围脑组织中occludin mRNA相对含量明显降低,在6～72 h持续维持在较低水平,与正常对照组比较差异均有统计学意义(P＜0.05). 结论 脑出血发生后,紧密连接蛋白occludin 的表达下降,这可能是脑出血发生后血脑屏障破坏及脑水肿发生发展的重要分子基础之一.     

局部亚低温对大鼠脑出血后基质金属蛋白酶-9及血脑屏障通透性的影响

目的探讨局部亚低温对大鼠自体血注入法脑出血模型基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)mRNA及蛋白表达的影响以及局部亚低温减轻脑出血后水肿的可能机制。方法雄性Wistar大鼠240只,随机分为脑出血(ICH)组和脑出血加局部亚低温(ICH H)组。每组分为对照、脑出血后6h、24h、72h、5d、7d共6个亚组,ICH H组于注血后立即给以4h的局部亚低温治疗,各亚组分别进行血脑屏障(BBB)通透性、脑水含量的检测以及应用RT-PCR及Western印记对MMP-9进行测定。结果ICH组大鼠脑内注血后6h开始出现脑组织水含量(P<0.01)及BBB通透性(P<0.05)的显著增加,二者在72h达到高峰,然后逐渐消退,ICH组MMP-9蛋白表达量与脑含水量和血脑屏障通透性呈正相关(r=0.88和r=0.96),ICH组MMP-9 mRNA表达量也与脑含水量和血脑屏障通透性呈正相关(r=0.78和r=0.85)。ICH H组大鼠脑组织水含量、BBB通透性以及MMP-9蛋白的表达与ICH组各时间点相比较,明显降低,而MMP-9 mRNA的表达与ICH组相比仅有轻度下降。结论脑出血后MMP-9的变化与BBB通透性和脑水肿密切相关,局部亚低温可以抑制脑出血后MMP-9蛋白表达的增加以及脑水肿的形成。提示局部亚低温可能通过影响MMP-9的变化来抑制脑出血后的水肿形成。     

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.     

亚低温对实验性大鼠脑出血后MMP-2/9表达的影响

目的 研究亚低温对脑出血后MMP-2/9表达及脑水肿的影响,探讨亚低温对出血性脑水肿的作用机制.方法 以立体定向技术在大鼠尾状核注射自体血制作脑出血模型;用冰块降温及白炽灯照射加温的方法调节体温;应用干湿重法测定脑水含量,并用免疫组化方法检测脑组织MMP-2/9表达.结果 脑出血模型大鼠血肿周围MMP-2/9的表达水平明显高于假手术组(均P<0.01);各个时间点亚低温组大鼠病灶侧血肿周围MMP-2/9的表达水平均明显低于常温对照组(P<0.01).同时,亚低温组脑水含量在各时间点与脑出血组比较明显下降(P<0.01).结论 亚低温能明显减轻脑出血后MMP-2/9的表达,亚低温可能通过抑制MMP-2/9的表达而减轻脑水肿.     

Epilepsy and anomalies of neuronal migration: MRI and clinical aspects

Neuronal migration disorders are the result of disturbed brain development. In such disorders, neurons are abnormally located. In diagnosing these conditions, magnetic resonance imaging is superior to any other imaging technique. This enables us to improve our knowledge of the clinical correlates of neuronal migration. With reference to migrational disorder, a retrospective study of all 303 patients with epileptic seizures referred for magnetic resonance imaging during a 3-year period was performed, 13 patients (aged 12-41, mean age 27) were identified. They represent 4.3% of the entire study group. Of the patients with known epilepsy, 6.7% and of the mentally retarded, 13.7% had migrational disorders. Four patients had schizencephaly as the dominant finding, one was classified as hemimegalencephaly, 2 had isolated heterotopias, and 6 had localized pachy- and/or poly-microgyria. The clinical pictures are complex. Ectopias of grey matter are recognised foci of epilepsy, but from an epileptological and a clinical viewpoint little attention has been given to these disorders. The present study shows that malmigration is not rare in epilepsy patients, especially not in the mentally retarded.     

Classification of methods in transcranial Electrical Stimulation (tES) and evolving strategy from historical approaches to contemporary innovations

Transcranial Electrical Stimulation (tES) encompasses all methods of non-invasive current application to the brain used in research and clinical practice. We present the first comprehensive and technical review, explaining the evolution of tES in both terminology and dosage over the past 100 years of research to present day. Current transcranial Pulsed Current Stimulation (tPCS) approaches such as Cranial Electrotherapy Stimulation (CES) descended from Electrosleep (ES) through Cranial Electro-stimulation Therapy (CET), Transcerebral Electrotherapy (TCET), and NeuroElectric Therapy (NET) while others like Transcutaneous Cranial Electrical Stimulation (TCES) descended from Electroanesthesia (EA) through Limoge, and Interferential Stimulation. Prior to a contemporary resurgence in interest, variations of transcranial Direct Current Stimulation were explored intermittently, including Polarizing current, Galvanic Vestibular Stimulation (GVS), and Transcranial Micropolarization. The development of these approaches alongside Electroconvulsive Therapy (ECT) and pharmacological developments are considered. Both the roots and unique features of contemporary approaches such as transcranial Alternating Current Stimulation (tACS) and transcranial Random Noise Stimulation (tRNS) are discussed. Trends and incremental developments in electrode montage and waveform spanning decades are presented leading to the present day. Commercial devices, seminal conferences, and regulatory decisions are noted. We conclude with six rules on how increasing medical and technological sophistication may now be leveraged for broader success and adoption of tES.     

Hepatic Considerations in the Use of Antiepileptic Drugs

Summary: Virtually all of the major antiepileptic drugs (AEDs) can cause hepatotoxicity, although fatal hepatic reactions are rare. The mechanisms, incidences, and risk profiles for such reactions differ from drug to drug. With carbamazepine and phenytoin, hepatotoxicity may be due to drug hypersensitivity. Although the profiles of patients at risk have not been well-defined for these two antiepileptic drugs, it would appear from reports in the literature that older adolescents and adults are at higher risk than children of developing serious or fatal hepatotoxicity. Once hepatotoxicity develops, mortality rates are 10–38% with phenytoin and 25% for carbamazepine. The risk profile for valproate fatal hepatotoxicity has been more clearly defined. Those at primary risk of fatal hepatic dysfunction are children under the age of 2 years who are receiving multiple anticonvulsants and also have significant medical problems in addition to severe epilepsy. The risk is considerably lower for patients over the age of 2 years on valproate monotherapy. In contrast to the risk profile with other AEDs, adults receiving valproate as monotherapy have the lowest risk of hepatotoxicity. Fatal hepatic dysfunction coincident with valproate may be the result of aberrant drug metabolism. Concomitant use of AEDs that induce microsomal P450 enzymes (e.g., phenytoin and phenobarbital) may enhance the production of a toxic metabolite, and hence the greater risk of hepatotoxicity with polypharmacy.     

Vascular Malformations and Epilepsy: Clinical Considerations and Basic Mechanisms

Summary: Vascular malformations (VMs) are associated with epilepsy. The natural history of the various VMs, clinical presentation, and tendency to provoke epilepsy determine treatment strategies. Investigations have probed the mechanisms of epileptogenesis associated with these lesions. Electrophysiologic changes are associated with epileptogenic cortex adjacent to VMs. Putative pathophysiologic mechanisms of epileptogenesis include neuronal cell loss, glial proliferation and abnormal glial physiology, altered neurotransmitter levels, free radical formation, and aberrant second messenger physiology.     

Neonatal Seizures: Problems in Diagnosis and Classification

Summary: The clinical identification of neonatal seizures is critical for the recognition of brain dysfunction; however, diagnosis is often difficult because of the poorly organized and varied nature of these behaviors. Current classification systems are limited in their ability to communicate motor, autonomic, and electroencephalo-graphic features of seizures precisely and to provide a basis for uniform effective diagnosis, therapy, and determination of prognosis. Recent investigations of neonates, utilizing bedside electroencephalographic/polygraphic/ video monitoring techniques, have provided the basis for improved diagnosis and classification of seizures in the newborn. These studies have demonstrated that not all clinical phenomena currently considered to be seizures require electrocortical epileptiform activity for their initiation or elaboration. In addition, the specific clinical character of the phenomena considered to be seizures, the clinical state of the infant, and the character of the EEG indicate the probable pathophysiological mechanisms involved and suggest probable etiologies, prognosis, and therapy. Similarities between animal models that demonstrate reflex physiology and neonates with motor automatisms and tonic posturing suggest that these clinical behaviors may not be epileptic in origin but, rather, primitive movements of progression and posture mediated by brainstem mechanisms. Although not all clinical behaviors currently considered to be neonatal seizures may have similar pathophysiological mechanisms, they are clinically significant because they all indicate brain dysfunction.     

Valproate Monotherapy in the Management of Generalized and Partial Seizures

Summary: For decades, therapeutic tradition has promoted the concept of polypharmacy in the management of epilepsy. In recent years, however, studies have shown that, for most patients, monotherapy can provide comparable or better seizure control than administration of multiple anticonvulsants, while diminishing the potential for adverse reactions, drug interactions, and poor compliance. Valproate is an important monotherapeutic agent that is highly effective in the control of idiopathic primary and secondarily generalized epilepsies, and partial seizures that do not generalize. Comparative studies have found that valproate is at least as effective as phenytoin and carbamazepine in the treatment of generalized and partial seizures. Given the similar efficacy, other factors such as pharmacokinetics and side effects may therefore determine anticonvulsant selection for monotherapy.     

Un nouveau cadre conceptuel de travail pour la psychiatrie

In an attempt to place psychiatric thinking and the training of future psychiatrists more centrally into the context of modern biology, the author outlines the beginnings of a new intellectual framework for psychiatry that derives from current biological thinking about the relationship of mind to brain. The purpose of this framework is twofold. First, it is designed to emphasize that the professional requirements for future psychiatrists will demand a greater knowledge of the structure and functioning of the brain than is currently available in most training programs. Second, it is designed to illustrate that the unique domain which psychiatry occupies within academic medicine, the analysis of the interaction between social and biological determinants of behavior, can best be studied by also having a full understanding of the biological components of behavior.     

Carbamazepine Efficacy and Utilization in Children

Summary: Carbamazepine is effective for preventing partial and generalized tonic-clonic seizures in children. Although absence epilepsies are more common in children than adults, an estimated 80% of children with epilepsy have seizure types or epilepsies that are potentially responsive to carbamazepine. The differential diagnosis of ictal staring is an especially important issue in children because absence and atypical absence seizures are more prevalent in children than adults. Age-related pharmacokinetic differences and drug interactions are major considerations in children. On average, children have higher clearance rates of carbamazepine, shorter half-lives, and higher ratios of carbamazepine-10, 11-epoxide to carbamazepine than adults. In addition, children with severe epilepsy are more likely to require multiple-drug therapy, which can lead to complex drug interactions. When carbamazepine is administered along with valproate, drug protein binding interactions can cause intermittent side effects.