袖套法建立大鼠肺移植模型的手术技巧

背景：建立大鼠肺移植动物模型是研究肺移植后病理生理过程及其治疗的基础和关键,但因其需要精细外科技术,难度较大,限制了这一模型的应用。 目的：探讨袖套法建立稳定的大鼠肺移植模型的手术操作技巧。 方法：纳入60只大鼠,采用改良的三袖套技术建立大鼠左肺原位移植模型30例。供肺植入血管、支气管套接吻合的技巧：受体肺静脉采用90°角分布的“4针悬吊法”,使塌陷的静脉血管开口完全张开且方向固定,减少受体静脉干短拙或强行套入导致血管壁撕裂的可能。受体肺动脉采用120°角分布的“3针悬吊”兜开,套接时应注意供肺套管的方向。最后支气管采用内支架吻合。计算各阶段手术时间及移植成功率。 结果与结论：30例大鼠肺移植手术成功率为90%,移植后存活率100%,左供肺摘取需时(3±1) min,供肺完成体外套管时间分别为：肺动脉(2.0±0.5) min、肺静脉(3±1) min、支气管(2.0±0.5) min,供受体动静脉和支气管套管吻合时间分别为：肺动脉(4±1) min、肺静脉(8±3) min、支气管(2.0±0.5) min,总手术时间(55±10) min。提示改良的三袖套技术建立大鼠肺移植模型操作简便,手术成功率高,可为大鼠肺移植缺血再灌注损伤研究提供稳定可靠的动物模型。     

改良Heron法的同种异体心脏移植模型建立

背景：啮齿类动物心脏移植模型分为工作模型和非工作模型,非工作模型相对于工作模型,手术时间短、成功率高,其中以Heron法最为显著。 目的：对Heron法建立大鼠心脏移植模型的手术方法进行改良,建立更合理、符合实验需要的心脏移植模型,以利于后续实验研究。 方法：对Heron法进行改良,将受体右颈外静脉套管插入供心右肺动脉,右颈总动脉套管插入供心主动脉,环扎固定,供心左肺动脉结扎,从而快速、高效地建立改良的心脏移植模型。观察大鼠移植成功率、并发症、总手术时间、受体血管套管准备时间,供心摘取时间,颈部移植时间及供心冷缺血时间。 结果与结论：单人操作连续进行正式实验25次,移植成功24次,成功率96%。全组无吻合口漏血、血液回流障碍,术后受体无死亡。每例手术总时间需50~60 min;受体血管套管准备时间为(18±3) min;供心摘取时间为(10±1) min;颈部移植时间为(6±1) min;冷缺血时间为(16±1) min。结果表明,改良Heron法建立心脏移植模型,无需显微外科操作,是一种冷缺血时间短,更合理、实用、稳定可靠和易于复制的器官移植研究动物模型。     

大鼠双侧供肾左侧原位肾移植模型的建立及肾功能监测

背景：大鼠肾移植模型是移植免疫耐受、免疫抑制及器官库开发的重要实验手段。 目的：建立实验动物使用效率高,重复性好的耐受、急性排斥、慢性排斥肾移植模型,及移植肾功能动态无创检测方法。 方法：封闭群Wistar大鼠用于解剖及预实验。采用改良左侧原位肾移植术,分次顺序使用供体左肾与右肾,动静脉及输尿管端端吻合,7 d后行右侧肾切除。DA、Lewis大鼠分别为供受体作为急性排斥组。F344、Lewis大鼠分别为供受体作为慢性排斥组。Lewis大鼠自体肾移植组作为移植耐受组。观测大鼠双侧肾脏应用解剖,供肾热缺血时间、冷缺血时间,肾移植手术成功率。检测分析肾移植模型生存曲线、蛋白尿水平动态变化。 结果与结论：解剖观测大鼠43例,灌注肾29例,行单侧原位肾移植21例。移植肾热缺血时间小于10 s,冷缺血时间(47.2± 3.7) min,成功率85.7%(18/21)。其中自体肾移植3例,急性排斥肾移植3例,慢性排斥肾移植12例。改良的单侧原位肾移植术简捷经济,分次顺序使用供体左肾与右肾,不干扰受体腹主动脉及下腔静脉血流,重复性好。结果提示,建立的近交系大鼠移植耐受、急性排斥、慢性排斥肾移植模型生存曲线、蛋白尿动态检测具有显著性差异。     

大鼠原位肾移植模型的移植技巧及并发症预防

背景：大鼠器官移植模型可重复性好，可稳定模拟临床缺血再灌注损伤、急性排斥反应及慢性排斥反应等情况，是免疫耐受以及新型免疫抑制剂开发不可缺少的组成部分。 目的：探讨稳定的大鼠原位肾移植模型的移植技巧及并发症的预防。 设计、时间及地点：显微外科操作实验，于2002-10/2008-03在中山大学附属第一医院动物中心、蒙特利尔大学实验中心及河南省动物中心完成。 材料：健康、雄性、清洁级Wistar大鼠、SD大鼠，分别为供、受体；Lewis大鼠、F344大鼠，分别为供、受体。 方法：在Fisher的移植方法的基础上进行改良，采用大鼠左侧原位肾脏移植，动、静脉及输尿管均采用端端吻合方式，实验在德国产Carl Zeiss手术显微镜下进行。 主要观察指标：监测移植后大鼠一般情况及尿量，大鼠尿量 < 1 mL认为移植肾脏功能差，活检确认原因。 结果：共实施大鼠原位肾移植500次。供体手术时间（15.5±2.1）min，受体移植时间（40.6±5）min，移植成功（移植后存活 > 5 d）率93.2%（466/500），一组慢性排斥模型大鼠达到长期存活（> 180 d）。 结论：施行多次肾移植可建立稳定、快速的大鼠原位肾移植模型，娴熟的动物外科解剖和精湛的显微外科技术是预防移植后并发症的先决条件。     

大鼠肾移植技术改进与分析

背景：大鼠肾移植模型是器官移植和肾脏相关疾病研究中常用的动物模型,目前已有多种建模方式,但这些模型在移植时间、移植效果等方面都存在各种问题。 目的：拟建立一种简便、安全、成功率高的单侧供肾大鼠肾移植模型。 设计、时间及地点：动物模型分析实验,于2008-03/06在复旦大学泌尿外科研究所完成。 材料：Wistar大鼠作供体,SD大鼠作为受体,各60只。 方法：切取供体左侧肾脏作为供肾,分别以供体腹主动脉、左肾静脉与受体腹主动脉、下腔静脉端侧吻合,供肾输尿管以5-0带针线牵引插入膀胱,吻合缝针破口完成尿路重建。肾包膜下切除受体原有双肾。统计移植时间和移植成功率。 主要观察指标：移植时间、热缺血时间、冷缺血时间、动静脉吻合和尿路重建时间、移植失败率及原因、受体大鼠存活率。 结果：手术时间100~140 min。其中供肾热缺血时间8~10 s,冷缺血时间40~60 min。移植成功率为85%,失败原因分别为吻合口狭窄伴血栓6.67%、吻合口出血5%,感染1.67%,不明原因1.67%。 结论：实验建立的建模方法简便、可靠,明显缩短热缺血及尿路重建的时间,减少相关并发症。     

经改进同种异体大鼠原位肾移植模型的建立

背景：大鼠肾移植模型是研究人体肾移植后排斥反应最好的方法,但因其需要精细外科技术,难度较大,限制了这一模型的应用。 目的：通过阅读大量相关文献,对多种造模手术方式和术中某些操作进行改进,建立稳定、可靠的大鼠原位肾脏移植模型显微外科手术方法。 设计、时间及地点：动物观察实验,于2008-09/11在天津市人民医院动物实验室完成。 材料：40只健康成年雄性SD大鼠,体质量250~300 g,随机选取20只作为肾移植供体,剩余20只为受体。 方法：①对供、受体进行肾血管游离时尽量采用钝性分离。②将供肾原位低温重力灌注,修剪、摘除后低温保存。③受体摘除左肾后行采用肾动、静脉端-端吻合,供体输尿管-受体膀胱浆肌层隧道术进行尿路重建,两点间间断缝合采用6针法吻合。术后小剂量环孢素抗急性排斥反应。 主要观察指标：通过对手术时间、术后死亡率及死亡原因分析,评价术式的合理性及可靠性。 结果：总手术时间(150±17) min,热缺血时间(15±2) s,冷缺血时间(55±5) min。3只于1周内死亡,其中麻醉意外1只,吻合口出血死亡1只,输尿管狭窄死亡1只,手术成功率为85%。术后第8天对存活受体鼠行剖腹探查,移植肾均颜色饱满,吻合口无栓塞、狭窄。 结论：通过对以往手术方式进行改进,采用肾动、静脉端-端吻合,供体输尿管-受体膀胱浆肌层隧道术进行尿路重建的术式建立了可靠的大鼠原位肾移植模型。     

改良大鼠减体积肝移植模型的效果分析

目的 探讨大鼠减体积肝移植术后肝脏再生实验研究中改良后的大鼠减体积肝移植模型的效果;方法 实验大鼠均为健康的SD大鼠,体重260g-280g,供体为雌性,受体为雄性,供、受体体重相差10g左右,一般为供体体重比受体体重轻;供体采用单人裸眼操作,在取肝的过程中即进行减体积操作;修肝时将套管柄置于门静脉和肝下下腔静脉的正前方,将幽门静脉结扎点外翻于套管外并置于套管柄的左侧,即肝脏的左侧;将右肾静脉结扎点外翻于套管外并置于套管柄的右侧,即肝脏的右侧;供肝套管完成后用灌注液对门静脉和肝下下腔静脉进行冲洗;然后以左膈静脉为标识点进行7-0无损伤血管缝线吊线;受体采用双人裸眼配合操作,肝上下腔静脉吻合时,左右固定位点采用“8”字形外翻缝合,后壁和前壁分别采用连续吻合,门静脉和肝下下腔静脉采用改良的双袖套法,胆管支撑管法建立大鼠减体积的稳定模型。 结果 供体手术时间为32±1.5min,修肝时间为6±1.6min,受体手术时间为40±2.5min,无肝期时间为14±2.3min;手术的成功率为92﹪,术后3天的存活率为90﹪,术后1周存活率为88.7﹪,术后2周存活率为88﹪;术后并发症较改良前明显较少,供肝的冷保存时间减少,差异有统计学意义。结论 改良后的大鼠减体积肝移植模型比较稳定和可靠,而且手术的成功率较高,术后的并发症较低,为更好的研究减体积肝移植术后肝脏再生的提供了有效的改良手段。     

一种可延长术后生存期的大鼠左肺原位移植模型制备方法*★

目的：目前袖套技术已广泛应用于肺移植的制备，但其远期效果不能肯定。实验拟改进大鼠左肺原位移植模型，重点解决动物移植术后存活时间的延长。 方法：①实验时间：实验于2007-03/07在上海市肺科医院动物实验室完成，动物实验方法符合医学伦理学要求。②实验材料及方法：取SD大鼠40只，沿用套管技术吻合肺动静脉，8/0带针锦纶丝线连续缝合支气管，建立20例同种异体大鼠左肺原位移植模型。③实验评估：术后监测动物胸部X射线及存活情况。麻醉处死动物后取出供体肺，制成4 μm厚切片并行苏木精–伊红染色，观察肺组织结构及细胞浸润情况。 结果：①共完成20例大鼠左肺原位移植手术，成功18例，失败2例，术后动物存活时间均超过3个月。②大鼠移植肺病理切片显示肺组织结构基本正常。 结论：该模型动物术后存活时间长，肺呼吸功能正常，适用于需长期存活的大鼠左肺原位移植实验研究。     

冷保存方法构建的大鼠肝移植模型

背景：肝移植是终末期肝病最为有效的治疗手段，移植后胆病是制约肝移植发展的主要障碍之一。 目的：建立稳定的大鼠冷保存肝移植模型，探讨冷保存对肝移植后肝脏胆管的影响。 方法：120只雄性SD大鼠随机分为4组，按照组内随机配对的原则，体质量相对较轻的大鼠做为供体，供肝置于4 ℃ UW液中分别保存2，8，16 h后行原位肝移植。在“两套袖法”基础上，以“支架法”建立动脉化大鼠原位肝移植模型，供受体肝总动脉采用改良“支架法”进行端端吻合，重建肝动脉血供。记录移植手术时间及移植成功率，并分别于移植后3，7 d检测血清转氨酶、总胆红素、碱性磷酸酶水平，同时观察组织病理学改变。 结果与结论：实验共完成55例次大鼠原位肝移植手术，手术成功率为93%。冷保存2，8，16 h组术后7 d存活率分别为100%(9/9)，83%(10/12)，73%(8/11)。随着冷保存时间的延长，肝功能及肝内胆管损伤加重，胆管组织病理学评分显示各组间差异有显著性意义(P < 0.05)。其中供肝冷保存16 h大鼠肝移植模型既有较高的手术存活率又有严重的胆管损伤，是研究冷保存对肝移植胆管病影响的较好模型。 主要观察指标：移植手术时间，移植成功率。分别于移植后3，7d检测血清转氨酶、总胆红素、碱性磷酸酶及组织病理分析。 结果：冷保存时间延长导致胆道功能严重损伤，胆管组织病理学评分显示各组间差异有显著性。 结论：供肝冷保存16 h大鼠肝移植模型是研究冷保存对肝移植胆管病影响的较好模型。     

30%小体积肝移植大鼠模型的手术技巧及改良

背景：活体肝移植显著缓解了供肝短缺矛盾,如何使供者献出的肝量最小,同时受者得到最大的临床受益成为了目前的研究热点。 目的：探索一种简便、稳定的小体积肝移植大鼠模型的建立方法。 方法：以Kamada“二袖套法”非动脉化原位肝移植SD大鼠模型为基础并参考国内外文献,制作30%小体积肝移植模型。实验分为Ⅰ组为体内肝叶减除;Ⅱ组为体外肝叶减除组;Ⅲ组为改良后小体积肝移植模型。主要观察供体手术各阶段时间;移植后并发症和生存情况。 结果与结论：改良后模型较改良前的供肝获取时间和肝叶减除时间均显著缩短(P < 0.05)。Ⅱ组的供肝冷缺血时间显著长于Ⅰ组和Ⅲ组(P < 0.05)。改良后的小体积肝移植模型手术成功率较改良前显著提高(P < 0.05),改良后7及14 d存活率均高于改良前模型(P < 0.05)。改良模型较改良前生存时间延长了9 d。改良模型的手术总并发症例次较改良前组显著较少(P < 0.01)。提示采用此改良方法能建立稳定的30%小体积肝移植大鼠模型,而且在操作简便、减少并发症、提高手术成功率和术后存活率等方面有显著优势。     

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.