实验性脑出血大鼠血肿周围VEGF、bFGF表达及血管新生的动态变化及重组人促红细胞生成素对其干预作用的研究

目的研究大鼠脑出血后血管内皮细胞生长因子(VEGF)、碱性成纤维细胞生长因子(bFGF)表达的动态变化及血管新生情况,以及重组人促红细胞生成素(rhEPO)对其的干预作用,探讨rhEPO对脑出血的可能保护机制。方法采用自体血脑内注入法建立脑出血动物模型。168只雄性SD大鼠随机分为正常组、假手术组、模型组、rhEPO治疗组。免疫组化法检测VEGF、bFGF、CD34的表达变化,用CD34的表达来反映血管新生情况。结果脑出血后3h即有VEGF表达增多,7～14d达高峰;bFGF的表达6h开始增多,72h达高峰;CD34+血管数12h开始增多,14d达高峰。rhEPO治疗组6h～21d时VEGF、bFGF表达比模型组高(P<0.05或P<0.01),且VEGF表达于72h～7d时提前达高峰;12h～21d时CD34+血管数高于模型组(P<0.01),且于7d时提前达高峰。结论rhEPO能上调脑出血后VEGF、bFGF的表达,促进脑出血后血肿周围新生血管生成。     

丁基苯酞对脑出血大鼠血管内皮生长因子、血管生成素-2蛋白表达及血管新生的影响

目的 通过观察大鼠脑出血后脑组织中血管内皮生长因子（vaseular endothelial growth factor, VEGF）、血管生成素-2（angiopoietin-2,Ang-2）蛋白在不同时间点的动态表达及不同剂量丁基苯酞干 预后其与新生血管数量的变化,探讨丁基苯酞对脑出血大鼠可能的神经保护作用及机制。 方法 通过自体血注入法制备SD大鼠脑出血模型并随机分为脑出血模型组、丁基苯酞低剂量组 及丁基苯酞中剂量组,以及对照的假手术组。丁基苯酞低、中剂量组大鼠分别予以丁基苯酞10 mg/kg、 25 mg/kg灌胃给药（每日2次）,假手术组及脑出血模型组大鼠则在相同时间用同等体积大豆油代替 灌胃。分别在术后1 d、3 d、7 d、15 d评估大鼠神经功能缺损,采用免疫组化法检测各时间点CD34的表 达并进行新生血管计数及血管场面积测定,检测各时间点VEGF、Ang-2蛋白的表达,测定脑出血大鼠 血肿体积。 结果 与脑出血模型组比较,丁基苯酞中剂量组在术后各时间点,低剂量组术后7 d、15 d VEGF蛋 白表达上调;丁基苯酞低、中剂量组术后各时间点An g-2蛋白表达均上调,血管计数均增多;丁基苯酞 低剂量组术后3 d的神经功能缺损评分较低,丁基苯酞中剂量组术后3 d、7 d、15 d的神经功能缺损评 分较低,上述差异均有统计学意义。丁基苯酞低、中剂量组与脑出血模型组,在术后7 d、15 d血肿体 积差异无统计学意义。 结论 丁基苯酞可以显著减轻脑出血大鼠的神经功能缺损,其作用机制可能与上调VEGF、Ang-2蛋 白的表达,增加脑出血血肿周围新生血管密度有关,同时未增加血肿增大的风险。     

G-CSF对脑出血大鼠血肿周围血管新生的作用机制研究

目的观察粒细胞集落刺激因子(G-CSF)对脑出血(ICH)大鼠血肿周围血管新生的影响,并探讨其机制。方法 63只SD大鼠随机分为假手术组、ICH组、治疗组,每组21只。利用立体定位仪,向SD大鼠右侧苍白球注入断尾获取的自体动脉血制备ICH模型,治疗组于造模1 h后腹腔注射重组G-CSF60μg/kg,假手术组、ICH组经腹腔注射等量等渗盐水。3组大鼠于术后6 h、24 h、48 h、72 h、7 d、14 d、21 d进行神经功能障碍评分,后处死大鼠,免疫组化检测血肿周围CD34+血管数、VEGF的表达。结果治疗组大鼠从24 h开始各时间点神经功能障碍评分较ICH组明显增加(P<0.05)。假手术组见少许CD34+血管表达;ICH组6 h开始CD34+血管表达增多,14 d达高峰;治疗组各时间点CD34+血管表达较ICH组明显增多(P<0.05),高峰提前至7 d。假手术组见少量VEGF表达;ICH组6 h即出现VEGF表达增多,7¹4 d达高峰;治疗组各时间点VEGF表达较ICH组明显增多(P<0.05),高峰提前至72 h14 d达高峰;治疗组各时间点VEGF表达较ICH组明显增多(P<0.05),高峰提前至72 h⁷ d。结论 G-CSF可上调ICH后VEGF表达,增加血肿周围新生血管生成,改善神经功能。     

MMP-9抑制剂对脑出血大鼠血管再生的作用

目的 探讨基质金属蛋白酶-9（MMP-9）对脑出血大鼠血管再生的影响.方法 通过自体血注射制作大鼠脑出血模型,脑出血后第1～7天给予MMP-9抑制剂（盐酸多西环素30 mg/kg,1次/d,灌胃）进行干预,第7天运用Longa评分法进行神经功能评分后,处死动物并取脑,免疫组化法检测CD34抗体标记的血管内皮细胞,测定血肿周围微血管密度（MVD）,并测定脑出血后血肿周围侧脑室附近的室管膜下区（SVZ） MMP-9的表达的灰度值,对结果进行统计学分析.结果 与对照组相比,模型组MMP-9表达增加、MVD增加,而干预组较模型组MMP-9表达减少,MVD增加,差异有统计学意义（P＜0.05）.干预组较模型组神经功能评分降低,差异有统计学意义（P＜0.05）.结论 脑出血急性期大鼠抑制MMP-9表达可能对其恢复期血管再生及神经功能修复有促进作用.     

重组人粒细胞集落刺激因子对脑出血大鼠脑水肿及血管再生的影响

背景：研究表明重组人粒细胞集落刺激因子可以减轻脑缺血后的脑水肿，并且可动员内皮前体细胞增加脑缺血区域新生血管生成，促进患者神经功能恢复。 目的：探讨重组人粒细胞集落刺激因子对大鼠脑出血后脑水肿及新生血管的影响。 设计、时间及地点：随机对照动物实验，于2006-03/11在泸州医学院中心实验室完成。 材料：健康雄性SD大鼠144只，随机分为假手术组、脑出血组、治疗组，48只/组。重组人粒细胞集落刺激因子为深圳新鹏生物工程有限公司产品。 方法：脑出血组与治疗组大鼠采用断尾取自体血方式建立脑出血模型，假手术组仅经注射点注入生理盐水。造模1 h后，治疗组大鼠腹腔注射重组人粒细胞集落刺激因子60 μg/kg。分别于干预后6 h，12 h，24 h，48 h，72 h，7 d时间点，每组各取8只大鼠，采用干湿重法测定大鼠组织脑含水量，采用SP、DAB显色法免疫组化检测CD34+血管的表达。 主要观察指标：脑组织含水量动态变化，CD34+血管免疫组化结果。 结果：与假手术组比较，脑出血组大鼠脑组织含水量显著升高(t=4.49，P < 0.05)，在48 h，72 h最明显；各时间点治疗组脑组织含水量均明显低于脑出血组(t=6.74，P < 0.05)。与假手术组比较，脑出血组CD34+血管数明显增加(t=3.42，P < 0.05)；治疗组CD34+血管数明显多于脑出血组(t=6.07，P < 0.05)，且在72 h，7 d时差异尤为明显。 结论：重组人粒细胞集落刺激因子作用于脑出血大鼠后，可减轻脑水肿程度，促进血肿周围新生血管生成。     

人重组粒细胞集落刺激因子对大鼠脑出血周围区新生血管的影响

背景：研究表明人重组粒细胞集落刺激因子可动员内皮前体细胞，增加脑缺血区域新生血管生成。 目的：观察人重组粒细胞集落刺激因子对大鼠脑出血周围区新生血管的影响。 设计、时间及地点：随机分组设计的动物对照实验，于2006-03/11在泸州医学院中心实验室完成。 材料：健康雄性SD大鼠72只，人重组粒细胞集落刺激因子。 方法：72只大鼠按随机数字表法分为3组，假手术组、脑出血组、治疗组，每组24只。断尾取自体血通过鼠脑立体定向仪注入鼠脑内制备脑出血模型，假手术组用生理盐水代替自体血。治疗组于制模后1 h腹腔注射人重组粒细胞集落刺激因子60 μg/kg。假手术组、脑出血组不做任何处理。 主要观察指标：于6，12，24，48，72 h，7 d 6个时间点检测血肿周围区CD34+血管的表达，每个时间点检测4只。利用内皮细胞的标志性抗原CD34变化了解微血管的生成情况，CD34抗原也越多，新生血管越多。 结果：脑出血组CD34＋血管数明显高于假手术组（P < 0.05）；治疗组CD34＋血管数明显高于脑出血组（P < 0.05），且在72 h增多明显，与7 d比较差异无显著性意义（P > 0.05），但与6，12，24，48 h比较差异有显著性意义（P < 0.05）。 结论：人重组粒细胞集落刺激因子能够促进脑出血后血肿周围新生血管生成。     

重组人粒细胞集落刺激因子对脑缺血再灌注大鼠的神经保护和血管再生作用

目的利用局灶性脑缺血再灌注模型,观察重组人粒细胞集落刺激因子(rhG-CSF)对脑缺血大鼠的神经保护和血管再生作用。方法健康雄性SD大鼠,随机分为假手术组,生理盐水对照组,rhG-CSF治疗组。线栓法制备大鼠大脑中动脉缺血(MCAO)模型,模型缺血90min时再灌注,治疗组予rhG-CSF 50μg/(kg·d),连续5d。每只大鼠均于治疗结束后分别进行神经功能缺损评分。采用免疫组化法检测CD105、VEGF的表达。结果 (1)病死率:对照组病死率为53.85%,rhGCSF治疗组为25%,差别有统计学意义(P0.05)。(2)神经功能评分:治疗后24h,对照组和治疗组较假手术组评分均明显上升,有显著性差异(P0.05)。治疗后7d和14d,治疗组较对照组神经功能恢复较好(P0.05)。(3)免疫组化结果:对照组及治疗组大鼠的CD105、VEGF阳性表达高于假手术组,治疗组的CD105、VEGF阳性表达要高于假手术组和对照组,差异有统计学意义(P0.05)。结论 rhG-CSF具有神经保护作用,能促进缺血区脑组织血管再生,其神经保护作用可能与促进血管再生有关。     

鼠神经生长因子对新生大鼠缺氧缺血性脑损伤后神经修复的作用

目的 探讨鼠神经生长因子(NGF)对新生大鼠缺氧缺血性脑损伤(HIBD)后神经修复的作用.方法 新生7日龄(SD)大鼠72只随机分成假手术组、模型组、治疗组,模型组腹腔注射生理盐水,治疗组腹腔注射NGF,采用免疫组化检测术后不同时间点(3 h、12 h、1 d、3 d、7 d、14 d)血管内皮生长因子(VEGF)及神经丝蛋白(NFP)的表达.结果 假手术组VEGF呈持续的低程度表达,HIBD后治疗组各时间点VEGF的表达均高于模型组,3 h～3 d治疗组的VEGF阳性区平均积分光密度(IOD)值大于模型组(P<0.05),且高峰提前至3 h,2组间7 d及14 d VEGF表达无统计学差异.HIBD后,3 h～1 d模型组和治疗组的NFP阳性区平均IOD值均显著低于假手术组(P<0.01),3 d、7 d及14 d治疗组NFP阳性区平均IOD值显著高于模型组(P<0.01).结论 NGF对 HIBD的神经修复机制之一,可能是通过上调VEGF及NFP的表达而实现.     

内皮祖细胞对局灶性脑缺血大鼠血管内皮修复作用的实验研究

目的 探讨内皮祖细胞对局灶性脑缺血后局部血管内皮的修复作用.方法 60只健康雄件SD大鼠随机分为对照组(5只)、假手术组(5只)和缺血-再灌注组(缺血组,50只),线拴法制备大腑中动脉闭塞模型,流式细胞术和免疫组织化学染色检测外周血 CD34和脑组织AC133表达水平.结果 再灌注后3 d,4d和7 d,缺血组大鼠外周血CD34表达水平降低且显著低于对照组和假手术组(均P<0.01);至再灌注后14d恢复至正常值水平,与对照组和假手术组比较差异无统计学意义(均P>0.05).再灌注后4 d,缺血组大鼠梗死侧大脑半球缺血半暗带区部分血管内皮细胞AC133表达阳性,主要集中于中、小血管内皮细胞胞质;其余各时间点各组大鼠AC133均表达阴性.结论 缺血-再灌注早期外周血CD34表达水平明显降低,梗死侧大脑半球缺血半晴带区部分血管内皮细胞AC133表达阳性,提示内源性内皮祖细胞可能参与局部血管内皮的修复.     

大鼠脑出血后MMP-9对血管生成影响

目的 探讨脑出血后MMP-9对血管生成的影响.方法 采用立体定向技术作大鼠脑出血模型,用免疫组化方法测定脑出血后CD34阳性新生微血管及MMP-9表达.结果 MMP-9阳性区平均灰度值高CD34阳性新生血管较多.结论 脑出血后MMP-9可能是促血管生成因子,强力霉素能通过调节MMP-9的表达与活性影响血管生成.     

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.     

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.     

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.     

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.     

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.     

Special Pharmacokinetic Considerations in Children

Summary: Pediatric patients have greater degrees of pharmacokinetic variability and unpredictability than adults. This variability results from the effects of pharmacogenetics, age and growth, prior and current comedication, and disease. Newborns with seizures have the least predictable dosage requirements, and their needs change as drug-eliminating mechanisms mature in the neonatal period. Infants have the highest relative capacities to eliminate antiepileptics of any age group and require the largest relative doses. In addition to age-related trends, children demonstrate the same drug-specific, pharmacokinetic phenomena that adults do, including nonlinear phenytoin elimination, nonlinear valproate binding, and autoinduction of carbamazepine. Intercurrent illness and drug interactions further modify the age-related pharmacokinetic patterns in children and make dosage requirements even more unpredictable. Recent studies have shown that febrile illness can affect drug elimination, sometimes decreasing drug levels by 50% or more. Intermittent treatment with benzodiazepines administered either orally or rectally can be an important adjunct and help minimize this type of problem for children with marginally controlled epilepsy. Intermittent benzodiazepines are also helpful for children who have febrile seizures and who need only occasional antiepileptic protection.