50例面肩肱型肌营养不良症的基因诊断与临床特征

目的对面肩肱型肌营养不良症(FSHD)患者进行基因诊断并总结其临床特征,以提高FSHD的诊断水平。方法以p13E-11为探针用EcoRⅠ BinⅠ双酶切的Southern杂交方法对50例FSHD患者进行基因诊断,并对其临床特征进行总结。结果50例FSHD患者的基因诊断结果为EcoRⅠ BinⅠ/p13E-11片段大小介于10～33．5 kb,平均(17．70±6．628)kb。家族遗传性患者符合常染色体显性遗传特点。FSHD多在青少年期慢性起病,病情缓慢进展；选择性侵犯面肌、肩带肌和上臂肌,部分患者逐渐累及盆带肌和下肢肌肉,患肌常不对称受累；血清CK水平正常或轻中度增高,肌电图示肌源性损害,肌活检呈肌病特征。结论以p13E-11为探针用EcoRⅠ BinⅠ双酶切的Southern杂交方法可对FSHD患者进行基因诊断,识别FSHD的临床特征以及进行基因诊断可提高FSHD的诊断水平。     

应用双酶切/Southern杂交方法诊断面肩肱型肌营养不良

目的：探讨面肩肱型肌营养不良（FSHD）的基因诊断方法。方法：抽取我院1997－2000年收治的37例FSHD患者的静脉血，常规抽提gDNA,以EcoR I/HindⅢ、EcoR I/Bln I 分别酶切gDNA,0.6%琼脂糖凝胶电泳分离，p13E-11探针Southern杂交，应用ImageMaster Total Lab v1.11分析软件判断杂交片段大小。结果：正常对照只检测到1个4q35来源的、大于33kb的EcoR I Hind Ⅲ／p13E-11片段，而所有FSHD患者中有33例检测到2个4q35来源的EcoR I HindⅢ/p13E－11片段，其中包含1个小于33kb的小片段；3例患者可检测到2个小于33kb的片段，但其中1个来自10q26；另1例散发性患者中检测到3个4q35型片段，且其中2个小于33kb。在1名无症状9岁男孩中检测到1个与其患病父亲一致的小片段，提示为症状前患者。结论：双酶切／Southern杂交的方法可用于中国人面肩肱型肌营养不良患者的基因诊断及症状前诊断。本文结果首次提示我国FSHD患者中也存在4q-10q的相互易位现象。     

实时荧光定量聚合酶链反应诊断面肩肱型肌营养不良

目的 建立面肩肱型肌营养不良(FSHD)的实时荧光定量PCR(FQ-PCR)检测方法.方法 常规酚-氯仿法抽提基囡组DNA,通过EcoR Ⅰ酶切及琼脂糖凝胶电泳,回收38 kb以下DNA作为模板,根据4号染色体上D424序列设计特异的引物和探针,对115例研究对象进行FQ-PCR检测,根据荧光曲线与阳性对照的比较判断结果.结果 16例已知EcoR Ⅰ片段大小的FSHD患者FQ-PCR检测结果为13例阳性,78名健康人检测结果除3例阳性外其余均为阴性,16例经临床诊断的新FSHD患者和5名高危者中分别有15例和3例检测结果为阳性.统计学分析FQ-PCR方法与传统印迹杂交方法(κ=0.765,P=0.002)及临床诊断(κ=0.844,P=0.000)之间的一致性,结果有统计学意义.结论 我们创建了以FQ-PCR技术对FSHD进行基因诊断的新方法.该方法能克服印迹杂交方法费时费力、有放射性污染的缺点,且能较好解决由于4q-10q易位及p13E-11探针结合部位缺失造成传统杂交基因诊断方法欠准确的问题,具有较好的临床应用价值.     

应用脉冲场电泳技术研究面肩肱型肌营养不良症的基因突变特征及进行基因诊断

目的 应用脉冲场电泳(PFGE)技术研究面肩肱型肌营养不良症(FSHD)基因结构突变特征，并进行基因诊断。方法 研究对象包括：110名健康对照，17例FSHD患者和23名家系成员。低熔点胶包埋法抽提基因组DNA，EcoRI、EcoRI／BlnI、XapI原位酶切，用脉冲电场凝胶电泳(PFGE)分离，p13E-11探针Southem杂交，曲线拟合法计算分析片段的长度。结果 17例患者均存在1条小于35kb的4q35 EcoRI致病片段；23名无症状家系成员中，发现2例患者的母亲分别携带25kb和28 kb的4q35 EcoRI短片段，因年龄较大，考虑为女性无症状顿挫型患者。另2例患者的女性同胞分别携带21．5 kb和31 kb的4q35 EcoRI短片段，因年龄较小，考虑为症状前患者。家族型和散发型患者均存在4q→10q型易位，各有2例，其中1例家族型患者为杂合易位；此外，在散发型患者中，发现2例男性患者存在体细胞嵌合现象。结论 我国FSHD患者存在易位、杂合、体细胞嵌合等复杂现象。应用PFGE技术对FSHD患者进行基因诊断具有准确性和可靠性。     

面肩肱型肌营养不良的分子遗传学研究进展

面肩肱型肌营养不良(facioscapulohumeral muscular dystrophy, FSHD)最早于1884年报道,是成人中最常见的肌营养不良,国外报道发病率约为1/20 000,呈常染色体显性遗传,但近年来发现不少新(当代)突变的散发病例.多于10～20岁起病,主要表现为面肌、肩胛带肌和上臂肌群进行性无力及萎缩,不同患者在起病年龄及临床表型方面有很大差异.1990年Wijmenga等[1]将其定位于4q35,从而使FSHD研究进入分子水平.近年来研究发现,95%的病例与4q35区的一个高度多态性EcoRI片段呈紧密连锁关系.此片段可用p13E-11探针通过Southern杂交检测,正常人为35～300 kb,病人则为10～34 kb.     

单分子荧光原位杂交技术在面肩肱型肌营养不良症精准诊断中的应用研究

目的探讨单分子荧光原位杂交(FISH)技术在诊断中国面肩肱型肌营养不良症(FSHD)患者的可行性。方法采用单分子FISH技术对37例临床拟诊FSHD患者4q A区域的D4Z4重复单元数进行检测和分析。结果 35例明确诊断为FSHD患者4q A区域的D4Z4重复单元数介于2~7,平均值为(4.229±1.031)。7例FSHD患者和2例临床拟诊FSHD患者的检测结果为:病例1、2、3来自同一个家系,为兄妹关系,4q A区域的D4Z4重复单元数分别为5、(6;32)和(5;32),片段长度分别为17.5 kb、(18.3;106.0)kb和(17.9;104.9)kb,D4Z4重复单元数在误差范围内,3例患者的变异可能遗传自其母亲;病例4的4q A区域D4Z4重复单元数为(2;22),片段长度为(6.6;71.9)kb;病例5、6的4q A区域D4Z4重复单元数分别为4和5,片段长度分别为11.7 kb和17.3 kb;病例7为女性,55岁,4q A区域D4Z4重复单元数为(5;68),片段长度为(18.1;224.0)kb,为本研究中年龄最大的女性患者;病例8、9为2例临床拟诊FSHD患者,4q A区域D4Z4重复单元数分别为(14;50)和33,片段长度为(44.6;164.8)kb和110.2 kb,4q A区域的D4Z4重复单元数均10。结论 FSHD患者存在高度家系间和家系内临床异质性。单分子FISH单次检测能同时准确分辨4种不同单倍型,即4q A、4q B、10q A和10q B的D4Z4重复单元数,可以明确检测致病相关的D4Z4单元重复数,误差在1 kb以内,能较精确地检测D4Z4单元重复数。因此,单分子FISH是现今精准的FSHD患者的临床分子诊断技术。     

家族性低钾型周期性麻痹的基因突变与临床特征

目的筛查家族性低钾型周期性麻痹相关基因突变位点,总结该病基因型和临床表型的相关性.方法应用聚合酶链反应（PCR）和DNA测序技术,对14个家族性低钾型周期性麻痹家系中的14例先证者进行候选基因CACNA1S、SCN4A、KCNE3的筛查,阳性者再对其家系中其他患者和健康亲属进行测序分析.结果14个家系中有3个家系其先证者存在已知的低钾型周期性麻痹相关突变（1个家系发生CACNA1S基因R1239H突变,2个家系发生SCN4A基因的R672H突变）.进一步对3个突变家系中4例其他患者和34名健康亲属测序分析发现,R1239H突变为完全外显率,R672H突变为不全外显率.同时还发现2种突变在发病年龄和乙酰唑胺的疗效等方面存在差异.结论中国低钾型周期性麻痹患者存在CACNA1S基因R1239H和SCN4A基因的R672H突变,2种突变的临床表型存在差异.     

面-肩-肱型肌营养不良症1型患者基因型与临床表型的相关分析

目的探讨面-肩-肱型肌营养不良症1型(FSHD1型)患者临床表型、基因型及二者之间的关联性。方法 2017年1月至2018年9月经单分子荧光原位杂交技术共确诊25例FSHD1型患者,对其临床资料、基因型及实验室检查结果进行回顾分析;按肌肉受累部位进行年龄校正临床严重程度评分(CSS),采用两独立样本的t检验比较不同D4Z4拷贝数患者年龄校正CSS评分差异,Spearman秩相关分析FSHD1型患者4qA的D4Z4拷贝数与年龄校正CSS评分及发病年龄之间的关联性。结果 D4Z4拷贝数为2～3个患者年龄校正CSS评分高于D4Z4拷贝数为4～6个者;D4Z4拷贝数目与年龄校正CSS评分呈负相关(r=-0.619,P=0.001),而与发病年龄呈正相关(r=0.516,P=0.012)。结论 FSHD1型患者4qA的D4Z4拷贝数与临床严重程度呈负相关,基因型对疾病严重程度具有提示意义,同时存在除基因型外可影响其临床严重程度的因素。     

精神分裂症与Xp11区HSU93305基因座的单体型及22 q11～13区相关基因的关联性研究

目的探讨Xp11区HSU93305基因座单体型、22 q11～13区α2肾上腺素能受体(A2αR)基因和儿茶酚-氧-甲基转移酶(COMT)基因多态性与精神分裂症的关联.方法分别提取59个中国汉族核心家系成员(母59名,父56名,精神分裂症患者59例)的DNA,采用基因扩增和限制性片段长度多态性技术,进行HSU93305基因座的单体型研究;选择其中56个父母均存活的家系进行A2αR基因和COMT基因多态性传递不平衡检验(TDT)的研究,并应用复等位基因TDT、基于单体型的单体型相对风险率(HHRR)检验等.结果 (1)Xp11区HSU93305基因座经Msp Ⅰ及Dra Ⅱ酶切后产生四种单体型D1M1,D1M2,D2M1,D2M2.父母组以单体型D2M1频率最高(57.7%);患者组也以D2M1传递率最高(59.0%),其次为D1M2(28.9%),D2M2传递率最低(1.2%).(2)经复等位基因TDT分析,精神分裂症与Xp11区HSU93305基因座相关联(χ2=9.28,v=3,P<0.05);与A2αR基因(χ2=1.09,v=1,P>0.05)和COMT基因(χ2=0.31,v=1,P>0.05)未见关联.(3)经HHRR检验,A2αR基因(χ2=1.21,v=1,P>0.05)和COMT基因(χ2=0.37,v=1,P>0.05)与精神分裂症亦未见关联.结论精神分裂症与HSU93305基因座相关联,其易感基因可能位于Xp11区;与A2αR基因和COMT基因多态性可能无关.     

先天性颅锁骨发育不全家系基因诊断探讨

目的探讨家族性颅锁骨发育不全家系的基因诊断方法。方法提取临床先天性颅锁骨发育不全两个家系中患者和健康成员外周血基因组DNA,PCR扩增RUNX2/CBFA1基因7个编码外显子及其侧翼内含子序列,分别进行正反向测序,测序结果与GenBank中的原始序列进行比对分析。根据人类基因突变命名规则确认测序发现的碱基突变。结果测序结果发现一家系中2例父子患者的RUNX2基因外显子1发生错义突变c.346T→A(W116R);另一家系中2位患者的RUNX2基因外显子3发生无义突变c.610A→T(K204X)。在2个家系中的健康成员和无关正常对照RUNX2基因DNA序列中没有发现上述突变。结论 RUNX2基因检测是对家族性颅锁骨发育不全家系进行基因诊断的准确有效方法,对其遗传咨询有重要意义。     

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.