重症肌无力患者糖皮质激素疗效与其受体α和β表达水平的关系

目的分析重症肌无力(MG)患者外周血单个核细胞(PBMC)胞质和胞核中糖皮质激素受体(GR)α和β表达与糖皮质激素(GC)疗效的关系,并探讨GC治疗MG疗效不同的机制。方法 收集24例MG患者和8名健康对照,根据GC疗效将MG患者分为GC敏感组、GC依赖组和GC抵抗组,每组各8例,应用Westernblot法检测MG患者GC治疗前后PBMC胞质和胞核蛋白GRtα和GRβ表达水平,并分析其与GC疗效的关系。结果正常情况下GRα主要表达于PBMC胞质中,GRβ主要表达于PBMC胞核中;GC抵抗组MG患者PBMC胞质中GRα表达水平降低(P0.05),而GC依赖组MG患者PMBC胞核中GRβ表达增高(P0.01)。GC治疗后,GC依赖组MG患者GRα的核移位率明显低于GC敏感组和GC抵抗组(P0.01)。结论 GC抵抗型MG与其PBMC胞质中GRα低表达相关,GC依赖型MG与其PBMC胞核GRβ高表达相关,并可能与由此所致的GRα核转位降低有关。     

重症肌无力患者外周血单个核细胞糖皮质激素受体α同型与激素疗效的关系

目的观察重症肌无力(MG)患者外周血单个核细胞(PBMC)糖皮质激素受体α同型(hGRα)含量或核转位率与激素疗效的关系。方法收集MG患者51例、其他神经免疫性疾病(ONID)患者16例及健康对照者11名,分别提取外周血单个核细胞(PBMC)细胞核和细胞质蛋白质,采用生物素-亲和素-酶联免疫吸附试验(ABS-ELISA)定量测定经体外地塞米松(DEX)作用前、后PBMC细胞质和细胞核内hGRα含量,并计算hGRα核转位率。应用临床相对记分法评价激素疗效。结果未经DEX处理时MG和ONID组PBMC细胞质hGRα含量均低于健康对照组(P<0.05),经DEX处理后MG组其hGRα含量明显低于健康对照组(P<0.01)。MG患者的性别、年龄、伴发胸腺疾病情况和hGRα含量以及hGRα核转位率间均无明显相关(P>0.05)。Osserman分型Ⅳ型MG患者hGRα核转位率低于其他分型者(P<0.05)。病程≥2年的MG患者hGRα核转位率明显低于病程≤6个月和病程6个月~2年的MG患者(均P<0.01)。MG患者PBMC的细胞核内hGRα含量和hGRα核转位率与激素治疗2周(n=51,r=0.913,r=0.907,P<0.05)和4周(n=47,r=0.929,r=0.911,P<0.05)的临床相对评分间均呈正相关。结论经DEX作用后PBMC细胞核内hGRα含量和激素诱导的hGRα核转位率与MG患者对激素的敏感性间呈正相关,两者可作为预测激素疗效的实验室指标。     

多发性硬化患者糖皮质激素受体蛋白和mRNA表达与激素疗效的相关性

目的 检测多发性硬化(MS)患者外周血单个核细胞(PBMC)中糖皮质激素受体(GR)蛋白与mRNA亚型:GRα mRNA、GRβ mRNA的表达,探讨GR蛋白和GRα mRNA、GRβ mRNA的表达与甲泼尼龙静脉冲击治疗(IVMP)疗效间的关系.方法 采用放射配体结合法测定20例缓解复发型MS(RRMS)、6例继发进展型MS(SPMS)及26名健康对照GR蛋白数量;采用半定量逆转录聚合酶链反应法测定GRα mRNA、GRβ mRNA的表达,使用扩展残疾状态量表(EDSS)评价IVMP疗效.结果 (1)IVMP前RRMS和SPMS患者GR蛋白数量分别为(3.8±0.2)×103位点/细胞和(1.6±0.2)×103位点/细胞,均显著低于健康对照组[(4.2±0.8)×103位点/细胞,P＜0.05].RRMS患者IVMP前GR蛋白数量与EDSS呈线性负相关(r=-0.441,P=0.015).(2)以磷酸甘油醛脱氢酶的表达作为内参照,RRMS患者GRα mRNA的表达水平(0.792±0.177)与健康对照组(0.805±0.158)差异无统计学意义,而SPMS患者的表达水平(0.315±0.129)明显低于健康对照(P＜0.05).(3)RRMS和SPMS患者PBMC中均有GRβ mRNA的表达,健康对照组未检出GRβ mRNA表达.(4)RRMS、SPMS患者GRβ mRNA的表达分别占各自GRα mRNA的43.98%±2.40%和140.01%±78.75%.结论 RRMS患者GRβ mRNA的低水平表达不影响IVMP疗效.SPMS患者GRα mRNA表达水平显著降低和(或)GRβ mRNA表达水平显著升高导致GC抵抗.     

重症肌无力患者外周血白细胞糖皮质激素受体的检测

目的探讨重症肌无力患者外周血白细胞糖皮质激素受体含量与重症肌无力发病的关系.方法用放免法测定重症肌无力患者血浆糖皮质激素水平,用放射配体结合法测定其外周血白细胞糖皮质激素受体含量.结果测定前2个月未曾接受糖皮质激素治疗的患者,其白细胞糖皮质激素受体平均值为(6799±2055)位点/细胞,与对照组(2839±1787)位点/细胞相比,位点数明显增高(P＜0.001).与患者的性别、年龄及疾病的临床类型无关.重症肌无力患者血浆皮质醇浓度为(458±170) nmol/L,正常对照组为(413±156) nmol/L,二者无明显差异(P＞0.05).结论重症肌无力患者外周血白细胞糖皮质激素受体含量增多,可能与重症肌无力发病有关.     

重症肌无力患者血清可溶性Fas水平变化及其临床意义

目的探讨重症肌无力(MG)患者血清可溶性Fas(sFas)水平变化及其临床意义。方法采用流式细胞术及酶联免疫吸附法(ELISA)分别测定45例MG患者(MG组)及40例对照者(对照组)外周血T细胞亚群分布和血清sFas水平,并对其中24例MG患者应用糖皮质激素(GC)治疗前后的血清sFas水平进行比较。结果(1)MG组外周血中CD8 T细胞百分率为(21.50±2.21)%,明显低于对照组[(27.75±3.00)%](P<0.01);CD4 /CD8 比值(1.92±0.26)明显高于对照组(1.48±0.16)(P<0.01);血清sFas水平[(3542.06±706.23)pg/m l]显著高于对照组[(2568.18±562.48)pg/m l](P<0.01)。(2)MG组全身型血清sFas水平为(3914.23±804.81)pg/m l,极明显高于眼肌型[(2797.72±592.83)pg/m l](P<0.001);病情中、重度患者明显高于病情轻度者(均P<0.005);预后良好者[(3254.69±661.73)pg/m l]显著低于预后不良者[(4178.38±841.65)pg/m l](P<0.001)。(3)24例MG患者经GC治疗后sFas水平[(2864.59±617.43)pg/m l]较治疗前[(3539.67±705.92)pg/m l]极显著降低(P<0.001)。结论MG患者血清凋亡抑制因素sFas水平增高,并与MG类型、病情及预后相关;GC治疗可以改善MG患者的免疫功能。     

microRNA-181c在重症肌无力患者外周血单个核细胞中的表达及其意义

目的探讨microRNA-181c(miR-181c)在重症肌无力(myasthenia gravis,MG)患者中的表达水平及其临床意义。方法收集2015年6月-2017年6月收治的22例MG患者和20例健康人的外周静脉血样,提取外周血单个核细胞(peripheral blood mononuclear cell,PBMC)中总RNA,采用实时定量PCR(quantitative real-time PCR,qRT-PCR)技术检测PBMC中miR-181c的相对表达水平,并分析miR-181c表达水平与患者的MG定量评分体系(quantitative myasthenia gravis score,QMG)的相关性。结果 MG患者PBMC中miR-181c的相对表达水平(0.303±0.150)较对照组(1.000±0.262)显著降低(P0.01),其中全身型患者(0.182±0.056)低于眼肌型患者(0.404±0.126)(P0.05),同时miR-181c表达水平与QMG呈负相关(R2=-0.359,P0.05)。结论 MG患者PBMC中miR-181c表达水平的降低可能与病情严重程度有关。     

重症肌无力患者对糖皮质激素的敏感性与糖皮质激素受体的关系

目的　了解重症肌无力(MG)患者对糖皮质激素(GC)敏感性与糖皮质激素受体(GR)的关系。 方法　观察10例MG患者及10名健康对照。采用 3　H-地塞米松( 3　H-DEX)放射配体法测定外周血单个核细胞(PBMC)GR。分别用植物血凝素(PHA)、髓鞘碱性蛋白(MBP)、乙酰胆碱受体(AChR)刺激PBMC， 3　H-TdR掺入法检测体外淋巴细胞增殖反应，地塞米松(DEX)抑制淋巴细胞增殖反应。 结果　正常对照及MG患者PBMC对PHA及MBP刺激的增殖反应差别不显著(P>,0 .05)。而MG患者PBMC对AChR的反应明显高于正常对照者(P<,0 .01)。DEX对MG患者PBMC特异性增殖反应的抑制率为42 .14%, 而对正常对照PBMC的增殖反应的抑制率为21 .62％，两组间有很显著区别(P<,0 .01)。GR数与DEX抑制率间有很良好的相关关系(r=0 .943, P<,0 .01)。 结论　MG PBMC GR数与体外DEX对PBMC 特异性增殖反应的抑制效应有良好的一致性。DEX对抗原特异性淋巴细胞增殖反应的抑制性可作为了解MG患者对GC敏感性的一项指标。     

多发性硬化患者糖皮质激素受体和甲基强的松龙疗效的关系

目的比较复发-缓解型多发性硬化(RRMS)、继发-进展型多发性硬化(SPMS)患者激素冲击治疗前、中、后外周血单核细胞(PBMC)中糖皮质激素受体(GR)蛋白含量和健康对照组的差异及其与甲基强的松龙(MP)疗效的关系。方法采用放射配体结合法测定45例健康成人、30例RRMS和6例SPMS患者PBMC中GR蛋白含量;使用扩展残疾状态量表(EDSS)评价MP疗效。结果(1)治疗前RRMS和SPMS患者的GR蛋白含量分别为(3765.83±1053.24)位点/细胞和(1599.67±448.79)位点/细胞,均低于健康对照组[(4249.09±616.10)位点/细胞](P<0.05);治疗中,RRMS和SPMS患者的GR蛋白含量分别为(3127.57±970.06)位点/细胞和(1219.67±346.27)位点/细胞,两者较治疗前均有下调(P<0.05);治疗后RRMS患者的GR蛋白含量上升至(4487.20±1473.18)位点/细胞,与健康对照组差异无统计学意义(P>0.05),而SPMS患者的GR蛋白含量上升至(3115.50±868.83)位点/细胞,但仍低于健康对照组(P<0.05)。(2)RRMS患者治疗后EDSS降至(2.70±1.80)分,与治疗前的(3.97±1.76)分相比差异有统计学意义(P<0.05);SPMS患者治疗后EDSS为(6.25±2.73)分与治疗前的(6.92±1.93)分之间差异无统计学意义(P>0.05)。(3)RRMS患者治疗前、后GR蛋白含量与同时期EDSS评分之间呈明显线性负相关(P<0.05)。结论GR蛋白含量的降低可能与MS的发生和发展有关,MP冲击治疗中症状一过性加重与GR蛋白含量下降相关,RRMS患者GR蛋白含量与MP的疗效相关。     

糖皮质激素受体α同型ELISA定量检测方法的建立

目的建立一种灵敏的定量检测人外周血单个核细胞(PBMC)细胞浆和细胞核内糖皮质激素受体α同型(GRα)的生物素-亲和素-酶联免疫吸附试验(ABS-ELISA)方法。方法分别提取人PBMC细胞浆和细胞核总蛋白,用GRα纯品或总蛋白浓度相等的细胞提取液包被酶标板,加入针对GRα的特异性兔抗人多克隆抗体(一抗)和山羊抗兔IgG(二抗),应用生物素-亲和素系统(ABS)进行放大,采用多因素棋盘滴定方法确定最佳实验条件,用GRα纯品建立ELISA标准曲线,定量检测GRα的含量。结果该研究确定应用0.1 mol/L碳酸盐缓冲液(pH 9.6)包被GRα纯品或细胞提取液,一抗浓度500 ng/100μL,二抗浓度400 ng/100μL,使用ABS放大系统,以TMB作为辣根过氧化物酶(HRP)的底物,于450 nm处检测吸光度值。该ABS-ELISA方法灵敏度为1μg/L,工作浓度范围1~100μg/L,批内变异系数(CV)为5.0%~6.9%,批间CV为7.1%~9.6%,不同操作者批内和批间的CV分别为4.7%和6.2%,均在实验允许范围内,可重复性良好。另外,该法无明显交叉反应,有一定特异性。结论该研究建立了一种特异性高、可重复性好、敏感、快速、简便的定量测定PBMC细胞浆和细胞核内GRα的新方法,对进一步探讨自身免疫病的激素疗效与患者GR的关系等方面具重要价值。     

MBP对PBMC产生IFN-γ和NO的影响

目的探讨T细胞非特异性活化在CNS脱髓鞘性疾病中的作用。方法分离实验性自身免疫性脑脊髓炎(EAE)易感性BALB/c小鼠外周血单个核细胞(PBMC),采用体外细胞培养方法在体外与碱性髓鞘蛋白(MBP)共培养,测定培养上清液中IFN-γ、NO水平。结果经MBP刺激的PBMC产生IFN-γ[(43.83±6.06)pg/mL]和NO[(180.76±20.75)μmol/L]明显增加,与对照组产生的IFN-γ[(28.52±2.18)pg/mL]和NO[(95.61±13.09)μmol/L]相比差异有统计学意义(P<0.01)。结论在CNS脱髓鞘性疾病发病过程中,活化的T细胞、单核细胞等分泌致炎细胞因子和其他有害物质增多。     

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.