5000/高氧液对大鼠肝移植的保护作用研究

目的 探讨高氧液对大鼠肝移植的保护作用。方法 40只SPF级Wistar 大鼠,随机分成2组,实验组实验高氧液灌注,对照组使用普通林格液灌注,比较两组灌注末、术后第1天、7天透明质酸、转氨酶、CD8+CD28-T细胞的变化,比较两组术后肝组织病理急性排斥评分。结果 术前两组无统计学差异,术后实验组透明质酸（87.9±6.3;65.4±5.1）、转氨酶（130.3±20.8;87.44±7.2）低于对照组（351.4±53.9;105.5±49.6）、（283.9±24.7;111.7±27.7）,CD8+CD28-T细胞（11.84±0.73; 14.76±0.62）高于对照组（6.22±0.2; 8.66±0.29）,（p<0.05）结论高氧液可减轻缺血再灌注损伤,对大鼠肝移植有一定的保护作用。     

不同缺血后处理方法对大鼠胰腺移植的保护作用及其机制

背景：缺血-再灌注损伤是胰腺移植后血栓形成和急性胰腺炎的主要原因之一,又是胰腺移植中不可避免的过程,会严重影响移植胰腺的功能。如何减轻缺血-再灌注损伤是目前移植外科的研究热点之一。 目的：观察不同缺血后处理方法对大鼠移植胰腺冷缺血再灌注损伤的保护作用。 方法：48只糖尿病SD大鼠随机数字表法分为为4组,对照组,缺血后处理1组(再灌注30 s缺血30 s 1次),缺血后处理2组(再灌注30 s/缺血30 s 3次)和缺血后处理3组(再灌注30 s/缺血30 s 6次),各组均行胰腺移植;48只建康SD大鼠为供体;检测各组大鼠再灌注前后血糖,再灌注后2 h移植胰腺组织中超氧化物歧化酶、丙二醛及髓过氧化物酶变化,TUNEL法检测移植胰腺组织细胞凋亡情况。 结果与结论：再灌注后缺血后处理组较对照组血糖降低(P < 0.01);缺血后处理2组较缺血后处理1组和缺血后处理3组血糖低(P < 0.05);再灌注后缺血后处理组较对照组移植胰腺组织中超氧化物歧化酶水平高(P < 0.01),丙二醛及髓过氧化物酶水平低(P < 0.01),缺血后处理2组较缺血后处理1组和缺血后处理3组超氧化物歧化酶水平高(P < 0.05)、丙二醛及髓过氧化物酶水平低(P < 0.05)。再灌注后缺血后处理组较对照组移植胰组织中细胞凋亡指数值低(P < 0.01),缺血后处理2组较缺血后处理1组和缺血后处理3组细胞凋亡指数值低(P < 0.05),提示缺血后处理对大鼠移植胰腺的缺血再灌注损伤具有保护作用,并可以减少移植胰腺缺血再灌注后的细胞凋亡,再灌注30 s/缺血30 s循环3次是最佳的大鼠移植胰缺血后处理诱导办法。     

丹参对大鼠移植肝血红素氧合酶1表达的影响*

背景：器官移植前使用丹参预处理能够保护组织缺血-再灌注损伤,改善移植器官存活率。 目的：观察含丹参的冷灌注液对同种异体大鼠移植肝脏中血红素氧合酶1表达的影响,以及对供体肝脏缺血-再灌注损伤的保护作用。 方法：将SD雄性大鼠随机分成UW液组(术中使用UW液灌注保存)、丹参+UW液组(术中使用丹参+UW液灌注保存)、ZnPP预处理组(移植前24 h腹腔内注射ZnPP,术中使用丹参+UW液灌注保存),建立稳定的大鼠同种异体肝移植模型。同时取10只正常大鼠作为正常对照。 结果与结论：丹参+UW液组和UW液组血清总胆红素、谷丙转氨酶、谷草转氨酶水平明显低于ZnPP预处理组(P < 0.01)。血红素氧合酶1mRNA及其蛋白在丹参+UW液组中较UW组表达更明显,在ZnPP预处理组中表达明显受到抑制(P < 0.05)。丹参+UW液组肝脏Suzuki标准评分明显低于ZnPP预处理组及UW液组(P < 0.05)。表明丹参能上调同种异体的大鼠移植肝脏中血红素氧合酶1 mRNA及其蛋白的表达,减轻供肝缺血-再灌注损伤,保护移植大鼠肝脏。     

丹参对原位肝移植大鼠供肝冷保存再灌注损伤的保护作用☆

目的: 研究表明，丹参对心、脑、肝等重要器官的缺血再灌注损伤有保护作用。制备大鼠异体原位肝移植模型，验证丹参对大鼠肝移植缺血再灌流损伤的保护作用。 方法：实验于2006-10/2007-08在南方医院中心实验室及动物实验中心完成，动物实验方法符合动物伦理学要求。①实验材料及分组：选用SD大鼠40只，按随机数字表法分为假手术组、模型对照组和丹参注射液组，假手术组8只，模型对照组、丹参注射液组各8对（供体与受体）。②实验方法：建立原位肝移植模型，在供肝灌注冷保存时，以4 ℃ 乳酸林格氏液为基液，丹参注射液组灌注保存液中加60 mL/L丹参注射液；模型对照组不加丹参。③实验评估：移植术后6 h处死各组大鼠取样，检测血清谷草转氨酶、谷丙转氨酶及乳酸脱氢酶活性；测定肝组织中丙二醛含量及超氧化物歧化酶、谷胱甘肽过氧化物酶活性，并对比观察移植肝病理形态学改变。 结果：模型对照组和丹参注射液组16只受体大鼠及假手术组8只大鼠全部进入结果分析，无脱失。①丹参注射液组和模型对照组移植肝再灌注后血清谷丙转氨酶、谷草转氨酶及乳酸脱氢酶活性均高于假手术组(P < 0.01)；丹参注射液组低于模型对照组(P < 0.01)。②丹参注射液组肝组织中丙二醛含量较模型对照组明显下降(P < 0.01)，超氧化物歧化酶和谷胱甘肽过氧化物酶的活性则明显升高 (P < 0.01)。③丹参注射液组较模型对照组肝组织肝细胞坏死程度减轻，炎性细胞浸润减少，肝组织再灌注损害程度减轻。 结论：丹参对原位肝移植肝脏的缺血再灌注损伤有保护作用，从而减轻氧自由基及脂质过氧化，保护细胞膜，改善肝功能。     

Genistein联合小剂量他克莫司在胰腺移植排斥反应中的作用

背景：有实验表明CXCR3拮抗剂Genistein可减轻胰腺移植大鼠的急性排斥反应。 目的：观察Genistein联合小剂量他克莫司在胰腺移植抗排斥反应中的作用。 方法：以Wistar大鼠为供体,SD大鼠为受体,建立胰腺移植模型,随机分成5组：对照组(未进行药物治疗)、大剂量他克莫司组、小剂量他克莫司组、Genistein组、Genistein+小剂量他克莫司组。术后第7天行病理学,肝肾功能,血清中CD3+、CD4+、CD8+T淋巴细胞、干扰素γ、白细胞介素2浓度检测。 结果与结论：与对照组、小剂量他克莫司组、Genistein组比较,Genistein+小剂量他克莫司组移植胰腺组织损伤减轻,淋巴细胞浸润减少,急性排斥反应减轻,血清CD3+、CD4+、CD8+ T淋巴细胞明显减少,干扰素γ、白细胞介素2水平降低 (P < 0.05);并且避免了大剂量他克莫司对肝肾功能的损害。说明Genistein联合小剂量他克莫司能有效减轻胰腺移植急性排斥反应而不增加肝肾毒性。     

无创伤双后肢缺血后处理对移植胰腺缺血再灌注损伤的远隔保护

背景：缺血预处理及缺血后处理是近年来提出减轻缺血再灌注损伤有效方法。 目的：探讨无创伤双后肢缺血后处理对移植胰腺缺血再灌注损伤的影响及机制。 方法：18只糖尿病SD大鼠数字表法随机分为3组,对照组仅行开腹术;缺血再灌注组仅行胰腺移植;缺血后处理组,移植前行非创伤性双后肢缺血后处理。 结果与结论：缺血再灌注组血糖和胰腺组织中丙二醛水平均高于缺血后处理组(P < 0.01)、而超氧化物歧化酶活性低于缺血后处理组(P < 0.01);与缺血后处理组比较,缺血再灌注组胰腺组织凋亡指数明显增高(P < 0.01)。结果提示,无创伤双后肢缺血后处理对大鼠移植胰的缺血再灌注损伤具有保护作用,机制可能与可通过减少超氧化物歧化酶失活,从而清除氧自由基以及减少胰腺细胞凋亡等有关。     

迟发缺血预处理低温保存肾移植供体中血红素加氧酶1的表达

背景：缺血预适应延迟反应通过诱导保护性蛋白增强组织对缺血再灌注损伤的耐受能力；血红素加氧酶1参与缺血预适应延迟保护作用。迟发缺血预处理对低温保存肾脏的作用及血红素加氧酶1是否参与其中尚不清楚。 目的：观察缺血预处理诱导血红素加氧酶1的迟发缺血预处理反应对低温保存肾脏移植供体的作用。 方法：雄性SD大鼠随机分入5组：空白对照组、低温保存组、缺血预处理组、缺血+低温组(n=12)；缺血+给药+低温组。各组大鼠均行右肾切除，预处理或假手术操作处理后24 h采用大鼠肾脏非循环离体灌注模型获取肾脏，分别于保存24，48，72 h取样。缺血+给药+低温组除上述处理外，还于原位低温灌注术前1 h接受1次血红素加氧化酶1抑制剂锡原卟啉腹腔注射。低温保存肾脏于各保存终点留取保存液，测定pH值和乳酸脱氢酶含量；切取1/2肾脏按照光镜要求制备标本送检；剩余1/2肾脏用于免疫印迹法测定血红素加氧酶1表达，比色法测定皮质Na-K-ATP酶活性、丙二醛和还原型谷胱甘肽含量；未保存肾脏仅通过免疫印迹法测定血红素加氧酶1的基础表达情况。 结果与结论：迟发缺血预处理诱导了肾组织血红素加氧酶1的表达，与单纯低温保存组相比保存24，48 h后，缺血+低温组保存液pH值、乳酸脱氢酶活性降低；肾脏组织Na-K-ATP酶活性、谷胱甘肽含量增加，丙二醛含量降低；同时点预处理组肾组织光镜形态学改变稍好于单纯低温保存组。给予血红素加氧酶1抑制剂后，这种保护作用消失。提示，迟发缺血预处理延长了肾脏低温保存时限，这可能与诱导血红素加氧酶1，增加组织抗氧化能力，减轻低温保存氧应激有关。     

丹参素干预体外低温保存大鼠肝脏血红素氧合酶1的表达

背景：研究发现丹参能够降低诱导型一氧化氮合酶mRNA 的表达，减少一氧化氮的产生，抑制肿瘤坏死因子、白细胞介素等炎性因子的分泌，具有抗氧化作用。丹参素作为丹参的重要单体成分，是否具有相同的作用? 目的：验证丹参素干预大鼠肝脏血红素氧合酶1的表达，以及对体外肝脏低温保存肝脏的保护。 方法：建立大鼠肝脏低温灌注保存模型。分为3组，对照组术前腹腔注射生理盐水，术中用乳酸林格液灌注；实验组术前腹腔注射生理盐水，术中用丹参素+乳酸林格液灌注；抑制剂组：术前腹腔注射锌原朴啉，术中用丹参素+乳酸林格液灌注。保存0，1，3，6 h，分别RT-PCR检测血红素氧合酶1mRNA及蛋白表达的情况，检测细胞线粒体钙离子含量及钙离子ATP酶活性，电镜观察各组肝细胞、线粒体形态改变，光镜观察肝细胞、肝小叶形态改变。 结果与结论：实验组肝血红素氧合酶1 mRNA及蛋白的表达水平明显比其他两组高(P < 0.05)。实验组的肝细胞线粒体钙离子含量明显较其他两组低，Ca2+-ATP酶活性较其他两组高。结果提示，丹参素灌注保存液可以诱导大鼠肝脏中血红素氧合酶1的过表达，延长肝脏低温保存时间。     

自制新型多器官保存液低温保存小型猪肾

背景：在前期实验的基础上,此次实验成功配制了含聚乙二醇的新型上海多器官保存液。 目的：以普通器官保存液为对照,验证自制含聚乙二醇的上海多器官保存液低温保存小型猪肾的效果。 设计、时间及地点：随机分组,对照实验观察,2006-05/2007-07在解放军器官移植研究所移植实验室完成。 材料：健康成年实验用巴马小型猪12头。自制新型新型上海多器官保存液,主要含聚乙二醇,相对分子质量20 000。pH7.43±0.10,渗透压825~900 kPa,UW液,购自美国Bristol-Myers Squibb公司。 方法：巴马小型猪肾分别以4 ℃器官保存液或上海多器官保存液原位灌注后离体保存24,48,72 h。保存终点测保存液pH值及乳酸脱氢酶含量,左肾取标本行组织学检查,右肾采用体外非循环猪肝/肾灌注系统灌注30,60,120 min测灌注液乳酸脱氢酶含量。灌注终点取肾皮质标本测丙二醛含量和超氧化物歧化酶活性。 主要观察指标：光镜、电镜观察肾冷保存后的形态学变化;灌注液及保存液乳酸脱氢酶含量;肾皮质标本测丙二醛浓度和超氧化物歧化酶活性;肾组织细胞凋亡指数。 结果：保存48,72 h后,器官保存液组保存液pH值高于上海多器官保存液组(P < 0. 05)。保存72 h终点保存液及再灌注30,60,120 min灌注液中乳酸脱氢酶活性器官保存液组高于上海多器官保存液组(P < 0.05)。同时间点两组肾组织光镜、电镜下形态学改变、肾皮质凋亡指数、超氧化物歧化酶、丙二醛含量差异均无显著性。 结论：新型上海多器官保存液对小型猪肾脏低温保存效果与普通器官保存液基本一致,对缺氧代谢器官细胞内酸中毒的缓冲能力以及对低温和缺血再灌注损伤的保护作用优于普通器官保存液。     

骨髓间充质干细胞对局灶脑缺血再灌注损伤的超早期干预

摘要：目的　探讨骨髓间充质干细胞(BMSCs) 对大鼠脑缺血再灌注损伤保护作用及机制。方法　雄性SD大鼠72只,以线栓法制成缺血再灌注模型,随机分为2组：溶剂对照组;BMSCs移植组。在缺血再灌注后1天、3天、6天分别行神经功能检测、TTC染色、免疫组化法检测Survivin、caspase-3表达,TUNEL法检测凋亡细胞表达。结果　与溶剂对照组比较,在梗死脑组织中移植骨髓间充质干细胞BMSCs后,可使大鼠神经功能有所恢复,在大鼠大脑中动脉局灶脑缺血90分钟再灌注3天及6天神经功能评分比较高,有统计学意义;TTC染色脑梗死体积百分比在缺血再灌注第3天、第6天较溶剂对照组分别减少2.13%和2.10%,有统计学意义。单纯BMSCs移植组比较对照组在缺血再灌注后1天、3天、6天缺血侧皮层Survivin表达增高、caspase-3表达降低,凋亡细胞减少,均有统计学意义。结论　脑缺血部位脑实质单纯 BMSCs 移植能够改善缺血后神经功能,减少脑缺血后梗死体积,可能通过增加脑缺血再灌注损伤部位Survivin蛋白的表达,降低凋亡相关蛋白Caspase-3表达,减少凋亡细胞数量发挥作用     

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.     

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.     

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.