目的 探讨上海地区抽动障碍(TD)患儿血清25-羟基维生素D水平的差异.方法 选择2018年5月至2020年5月上海市儿童医院(以下简称"我院")神经内科门诊就诊的5～14岁TD患儿2001例,及我院同期体检健康的5～14岁正常儿童2670名.比较TD患儿与正常儿童血清25-羟基维生素D的差异,同时比较不同特征的TD患儿的血清25-羟基维生素D的差异.进一步将TD患儿按照每2岁(24个月)一个组段,划分为5个年龄组,包括5～6岁组(655例),7～8岁组(237例),9～10岁组(610例),11～12岁组(342例)和13～14岁组(157例),比较五组25-羟基维生素D的差异.结果 TD患儿血清25-羟基维生素D缺乏程度高于正常儿童,差异有统计学意义(P<0.05).不同居住地、户外活动时间、核心家庭成员性格、饮食习惯、学习压力的TD患儿血清25-羟基维生素D缺乏程度比较,差异有统计学意义(P<0.05).9～10岁组、11～12岁组及13～14岁组血清25-羟基维生素D缺乏程度高于5～6岁组和7～8岁组,差异有统计学意义(P<0.05).结论 上海地区TD患儿明显存在25-羟基维生素D缺乏,不同特征的TD患儿25-羟基维生素D缺乏程度不同,9岁以上的青少年缺乏最为显著.

Aristaless-related homeobox (ARX)-related disorders are recessive X-linked intellectual disability disorders. We encountered a patient with a hemizygous mutation (c.1507_1508del) showing intellectual disability, early-onset epileptic encephalopathy and Ohtahara syndrome. The patient had female genitals, but an XY karyotype. We established an induced pluripotent stem cell (iPSC) line from the peripheral blood mononuclear cells (PBMCs) of a six-month Chinese child with a hemizygous mutation (c.1507_1508del) in ARX. The PBMCs were reprogrammed with Sendai viral vectors. The iPSCs showed stable amplification, pluripotency-related gene expression, and trilineage differentiation potential. Karyotype analysis of the iPSCs showed 23 pairs of chromosomes with normal structure and sex chromosome is XY.

The gene encoding collagen like tail subunit of asymmetric acetylcholinesterase (COLQ) is responsible for the transcription of three strands of collagen of acetylcholinesterase, which is attached to the endplate of neuromuscular junctions. Mutations in the COLQ gene are inherited in an autosomal-recessive manner and can lead to type V congenital myasthenia syndrome (CMS), which manifests as decreased muscle strength at birth or shortly after birth, respiratory failure, restricted eye movements, drooping of eyelids, and difficulty swallowing. Here we reported three variants within COLQ in two unrelated children with CMS. An intronic variant (c.393+1G>A) and a novel missense variant (p.Q381P) were identified as compound heterozygous in a 13-month-old boy, with the parents being carriers of each. An intragenic deletion including exons 14 and 15 was found in a homozygous state in a 12-year-old boy. We studied the relative expression of the COLQ and AChE gene in the probands' families, performed three-dimensional protein structural analysis, and analyzed the conservation of the missense mutation c.1142A>C (p.Q381P). The splicing mutation c.393+1G>A was found to affect the normal splicing of COLQ exon 5, resulting in a 27-bp deletion. The missense mutation c.1142A>C (p.Q381P) was located in a conserved position in different species. We found that homozygous deletion of COLQ exons 14–15 resulted in a 241-bp deletion, which decreased the number of amino acids and caused a frameshift translation. COLQ expression was significantly lower in the probands than in the probands' parents and siblings, while AChE expression was significantly higher. Moreover, the mutations were found to cause significant differences in the predicted three-dimensional structure of the protein. The splicing mutation c.393+1G>A, missense mutation c.1A>C (p.Q381P), and COLQ exon 14–15 deletion could cause CMS.

Zellweger spectrum disorder (ZSD) is a heterogeneous group of autosomal recessive disorders characterized by a defect in peroxisome formation and attributable to mutations in the PEX gene family. Patients with ZSD have profound neurologic impairments, including seizures, severe retardation, and dysmorphic features, and poor prognosis. Currently, there is no specific, effective treatment. Here, we investigated the effects of allogeneic hematopoietic stem cell transplantation (allo-HSCT) on PEX1-related ZSD. The suspected clinical proband was first diagnosed at the Department of Neurology of our hospital. The proband died soon after diagnosis, and his family was studied. We found that a brother had the same genetic alterations, and he was diagnosed with Infantile Refsum disease (IRD) as the mildest form of ZSD. We implemented treatment with allo-HSCT, at the request of the child's parents. After transplantation, we observed significant improvements in the clinical manifestations, very-long-chain fatty acids, and brain MRI. The patient has recovered well and not showed any abnormal clinical manifestations after 2 years of follow-up. We have achieved satisfactory short-term results in the treatment of ZSD-IRD with allo-HSCT. Long-term follow-up and observation will be performed to determine the long-term prognosis.

Mental retardation, X-linked 21/34 (MRX21/34), is a rare intellectual disability disease caused by mutations in the IL1RAPL1 (Interleukin-1 Receptor Accessory Protein-Like 1) gene. Using Sendai virus-mediated reprogramming, we established an induced pluripotent stem cell (iPSC) line from PBMCs collected from a ten-year-old boy with MRX21/34. The iPSCs showed stable amplification, expressed pluripotent genes, displayed a normal karyotype, and had characteristics of trilineage differentiation potential in an in vitro differentiation assay.

The RNA polymerase II transcription subunit 12 homolog (MED12) is a member of the mediator complex, which plays a critical role in RNA transcription. Mutations in MED12 cause X-linked intellectual disability and other anomalies collectively grouped as MED12-related disorders. While MED12 mutations have been most commonly reported in male patients, we present the case of a 1-year-old girl with clinical characteristics similar to MED12-related disorders. To explore the clinical characteristics of the condition and its possible pathogenesis, we analyzed the patient's clinical data; genetic testing by whole-exome sequencing revealed a de novo heterozygous mutation (c.1249-1G > C) in MED12. Further cDNA experiments revealed that the patient had an abnormal splicing at the skipping of exon9, which may have produced a truncated protein. qPCR showed decreased MED12 gene expression level in the patient, and an X-chromosome inactivation test confirmed a skewed inactivation of the X-chromosome. The lymphoblast transcription levels of the genes involved in the Gli3-dependent sonic hedgehog (SHH) signaling pathway, namely, CREB5, BMP4, and NEUROG2, were found to be significantly elevated compared with those of her parents and sex- and age-matched controls. Our results support the view that MED12 mutations may dysregulate the SHH signaling pathway, which may have accounted for the aberrant craniofacial morphology of our patient.

Mitochondrial DNA depletion syndrome-13 (MTDPS13) is a rare autosomal recessive mitochondrial disease caused by mutations in the FBXL4 (F-box and leucine-rich repeat protein 4) gene. Using Sendai virus-mediated reprogramming, we established an induced pluripotent stem cell (iPSC) line from PBMCs collected from a one-year-old female patient with MTDPS13. The iPSCs were stable during amplification, expressed pluripotent genes, maintained a normal karyotype, and showed characteristics of the three germs layers in an in vitro differentiation assay.

Allan–Herndon–Dudley syndrome (AHDS) is a rare, X-chromosome-linked inherited disorder that affects brain development and is caused by a mutation in SLC16A2. Herein, we generated an induced pluripotent stem cell (iPSC) line from the peripheral blood mononuclear cells of a one-year-old male infant with AHDS using Sendai-virus-mediated reprogramming. These iPSCs exhibited stable amplification, expressed pluripotent markers, and differentiated spontaneously into three germ layers in vitro. Additionally, this iPSC line was found to maintain a normal karyotype and retain the pathogenic mutation in SLC16A2, facilitating the study of disease mechanisms and development of new therapies of AHDS.

Background Diagnoses of vanishing white matter disease (VWMD) were difficult due to variable clinical features, severity, age of onset and wide range of mutations in eIF2G genes which cause VWMD. This study reported two novel mutations in eIF2B genes associated with VWMD to and expand our understanding of VWMD. Case presentation Relevant data from clinical diagnoses and genetic mutational analyses in two Chinese female patients with sporad­ic VWMD were collected and analyzed. Protein structure/function was predicted. The identity of biological parents was confirmed based on variants called from the next-generation sequencing (NGS) data. Compound heterozygous mutations, c.254 T>A (p.Val85Glu), and c.597+2delT in the EIF2B2 gene, c.545C>T(p.Thr182Met) and c.1340C>T(p.Ser447Leu) in the EIF2B5 gene were detected in the two patients. Further phenotype investigation of both patients enables the diagnosis of the vanishing white matter disease .Three missense mutation c.254 T>A (p.Val85Glu) in the EIF2B2 gene, c.545C>T(p.Thr182Met) and c.1340C>T(p.Ser447Leu) in the EIF2B5 gene have been found and predicted to be deleterious. All the three mutation causes hydrophobicity and stability changes of proteins, and all the mutations were localized in conserved sequences. One novel mutation of c.1340C>T(p.Ser447Leu) in the EIF2B5 gene ,and two other known mutations. The iterative threading assembly refinement (I-TASSER) server generated three-dimensional (3D) atomic models based on protein sequences from the novel missense mutation of c.1340C>T(p.Ser447Leu) in the EIF2B5 gene, which showed that the protein structure changed . The novel mutation c.597+2delT in the EIF2B2 gene may cause the splice site to disappear. We also analyzed mutations in missense mutations that cause VWMD and found that most of the t Pathogenic sites are localized in conserved NT and I-patch homology regions and the catalytic domain of EIF2Bε. Conclusions This study expands the spectrum of genotypes and phenotypes of VWMD and provides new insights into the molecular mechanism of VWMD and aide the acute diagnosis and treatment of VWMD.

Mediator complex subunit 12 (MED12)-related disorders are recessive-X-linked intellectual disabilities present primarily in male patients. We came across a female patient with a heterozygous mutation (c.1249-1G > C) related to MED12-related syndrome. MED12 expression was significantly lower than that in her parents, and another X chromosome was inactive. We established an induced pluripotent stem cell (iPSC) line from peripheral blood mononuclear cells (PBMCs) of a 1-year old Chinese girl with a heterozygous mutation (c.1249-1G > C) in MED12. PBMCs were reprogrammed using nonintegrative Sendai viral vectors. The iPSCs showed stable amplification, pluripotency-related gene expression, trilineage differentiation potential, and a normal karyotype.

目的 对临床诊断疑似为脊肌萎缩症(SMA)的患儿进行运动神经元生存基因(SMN)缺失分析,通过基因诊断对SMA进行确诊.方法 收集2015年1月至2016年12月上海市儿童医院新生儿科及神经内科就诊的疑似SMA患儿29例,患儿就诊年龄11 d～14岁.采用多重连接探针扩增(MLPA)技术对上述患儿外周血DNA样本进行基因诊断.结果 15例存在SMA致病突变,疑似患儿的SMA基因诊断率为51.7％(15/29).其中8例为SMN1基因外显子7和外显子8纯合缺失;3例为SMN1基因外显子7和外显子8纯合缺失,SMN2基因外显子7拷贝数增加;4例为SMN1基因外显子7纯合缺失,外显子8杂合缺失.另外14例存在不直接致病的突变.按照SMA国际诊断标准,15例确诊患者中Ⅰ型5例,Ⅱ型5例,Ⅲ型5例,均占33.3％.其中Ⅰ型的5例患儿均为SMN1基因外显子7、8纯合缺失,仅有1例存在SMN2基因外显子7拷贝的轻度增加.结论 MLPA技术灵敏度较高,可作为SMA疑似患者首选的诊断方法,为SMA的明确诊断提供依据.

Background: This study aimed to clarify the diagnosis and expand the understanding of dopa-responsive dystonia (DRD). Material/Methods: Relevant data from clinical diagnoses and genetic mutational analyses in 3 Han Chinese patients with sporadic DRD were collected and analyzed. Protein structure/function was predicted. Results: One novel mutation of c.679A>G (p.T227A) in GCH1 and 3 known mutations of c.457C>T (p.R153X), c.739G>A (p.G247S), and c.698G>A (p.R227H) in tyrosine hydroxylase (TH) have been found and predicted to be damaging or deleterious. All of the mutations were localized in conserved sequences. The iterative threading assembly refinement (I-TASSER) server generated three-dimensional (3D) atomic models based on protein sequences from the novel nonsense mutation of c.679A>G (p.T227A) in GCH1, which showed that residue 227 was located in the GCH1 active site. Conclusions: Patients carrying different non-synonymous variants had remarkable variation in clinical phenotype. This study expands the spectrum of genotypes and phenotypes of DRD in the Han Chinese ethnicity, provides new insights into the molecular mechanism of DRD, and helps the diagnosis and treatment of DRD.

RATIONALE:Congenital myasthenic syndrome (CMSs) are a group of rare genetic disorders of the neurological junction, which can result in structural or functional weakness. Here, we characterized a case of CMS in order to clarify the diagnosis and expand the understanding of it. The molecular diagnosis had implications for choice of treatment and genetic counseling.PATIENT CONCERNS:A 3-year-old male patient with CMS had ptosis and limb weakness for 2 months after birth. Clinical course and electrophysiological, imaging, and genetic findings were assessed. Protein structure/function was predicted. A novel mutation of c.295C>T (exon 4) and another known mutation of c.442T>A (exon 5) were found in CHRNE. Both mutations localized in conserved sequences. The c.442T>A (p.C148S) missense mutation in CHRNE was predicted to be damaging/deleterious. The iterative threading assembly refinement (I-TASSER) server generated vastly different 3-dimensional (3D) atomic models based on protein sequences from wide-type and novel nonsense mutation of c.295C>T (p.R99X) in CHRNE.DIAGNOSES:The diagnosis of CMS with CHRNE mutations in Han Chinese was confirmed.INTERVENTIONS:The patient was given prednisone (10 mg, once daily, taken orally) and pyridostigmine (15 mg, three times a day, taken orally).OUTCOMES:The patient had a moderate response to prednisone and pyridostigmine.LESSONS:We expanded the genotype and phenotype of CMS with CHRNE mutations in Han Chinese and provided new insights into the molecular mechanism of CMS and help to the diagnosis and treatment of CMS.

目的 探讨中国汉族多发性硬化(MS)病儿的临床特点.方法 对2013年6月-2016年5月我院住院及门诊诊断的15例多发性硬化病儿的临床特征及治疗情况进行回顾性分析.结果 本组15例病儿中,男6例,女9例;起病年龄2～12岁,平均起病年龄6岁.急性或亚急性起病;5例病儿首发症状为视力障碍、复视及视物模糊.颅脑MRI检查均异常,病灶主要分布在脑室周围、基底节、脑干和小脑区域.应用甲基强的松龙联合丙种球蛋白治疗,病儿均经过复发-缓解的过程.其中3例复发超过4次,再次用激素治疗仍然有效.结论 儿童期MS以视神经炎起病多见,复发频率高,急性期激素和丙种球蛋白治疗有效.

目的 探讨生酮饮食疗法治疗线粒体脑肌病(MELAS)的临床疗效,提高临床医生对生酮饮食及线粒体疾病的认识.方法本文对2例明确诊断为线粒体脑肌病(MELAS)伴有癫痫的患儿进行生酮饮食并随访,对比生酮饮食前后患儿的临床症状、头颅影像学、脑电图等方面的临床变化,并分析生酮饮食对改善线粒体脑肌病临床症状的作用机制.结果1例MELAS患儿临床乳酸持续升高、反复发作性头痛伴呕吐、癫痫发作、影像学监测出现新的病灶.经过生酮饮食1月后,头痛呕吐症状消失,乳酸恢复正常,影像学未见明显改善.另1例MELAS患儿临床反复抽搐发作,生酮10个月,无抽搐发作,影响学提示颅内异常信号影较前明显缩小.结论生酮饮食可以作为治疗线粒体病的辅助治疗的方法,是安全有效的,尤其是降低血乳酸水平及减少临床的抽搐发作.

Background/aim: Multiple sclerosis (MS) is a chronic and debilitating inflammatory autoimmune disease of the central nervous system (CNS) that affects the myelinated axons in the CNS. Incomplete remissions occur more commonly with increasing duration of disease. Intravenous immunoglobulin (IVIg) has various functions as an immune modulator via macrophage activation. Clinical trials of immunoglobulin demonstrated remarkable clinical effects in several types of MS, especially in relapsing-remitting type. It is an approved method for the treatment of relapsing-remitting MS that can be used as a supportive therapy. Our study involves the case of a ten year old female patient with relapsing-remitting MS. This study was undertaken to examine the effects of IVIg used almost every 6 months in a patient with relapsing-remitting MS. Results: This case study demonstrated that treatments of IVIg used almost every 6 months in a patient with relapsing-remitting MS have potent therapeutic actions with early beneficial responses. Conclusion: IVIg used almost every 6 months shows a potential positive therapeutic treatment for relapsing-remitting MS and more large-scale clinical studies are required.

Objective To investigate the clinicalmanifestation,monitoring and therapeutic measure of severe enterovirus 71 ( EV71 ) infection in children.Methods Forty-five cases of severe EV71 infection were admitted in our PICU from May 2010 to Sep 2011.The vital sign and arterial blood pressure,central venous pressure,mixed venous oxygen saturation,dynamic non-invasive heart function and urine volume were monitored.Forty-five cases were divided into 3 stages according to clinical manifestation:( 1 ) nervous system involvement stage; (2) respiratory system involvement stage; ( 3 ) circulatory system involvement stage ( compensation and decompensation).We adopted individualized remedy measure according to different stages.Results In 45 cases,38 cases discharged from hospital,the cure rate was 84.4％.Among all the 38 cases,nervous system involvement was found in 19 cases,respiratory system involvement was found in 12 cases,circulatory system involvement was found in 7 cases.Seven cases died,who had circulation failure.Conclusion We should identify severe EV71 infection early.Positive control of high fever,appropriate liquid treatment,control of high blood pressure,early respiratory support,preventment of circulation failure are the key measures for treatment.Individualized monitoring and treatment are effective in children with severe EV71 infection.

Objective To investigate the efficacy of continuous veno-venous hemodialysis/filtration (CVVHDF) in children with acute liver failure.Methods Five cases of acute liver failure in children were treated in Shanghai Jiao Tong University affiliated Children’s Hospital from May 2009 to May 2010.Two cases were male and 3 cases female,aged from 10 months to 5 years old.The duration of disease on admission was 3 d to 14 d.Three cases of acute liver failure were caused by medicine(paracetamol),1 case was caused by severe sepsis,and 1 case was caused by pylephlebitis.The hepatic encephalopathy class of the patients were: 1 case (grade 2),two cases (grade 3),two cases (grade 4).Besides protecting liver function drugs,we used CVVHDF to treat the patients and closely monitored the vital sign,liver function and complications.Results The time of CVVHDF initiated was 2 h to 24 h after admission.The total duration of CVVHDF was 24 h to 144 h.Three cases recovered,1 case withdrew and 1 case died.Twelve hours after CVVHDF,ALT decreased from (3888.76±2373.60)U/L before CVVHDF to (3284.80±1974.80)U/L and continued to decrease (F = 3.58,P < 0.05).Blood ammonia decreased from (209.00±53.61)μmol/L to (158.80±60.93)μmol/L (F = 3.75,P < 0.05),and TBIL improved after 12 h～48 h,but there was no statistical difference compared with those at the beginning of CVVHDF (F=0.23,P > 0.05).The blood pressure was stable during the course of treatment.Conclusion CVVHDF treatment for acute liver failure can improve clinical symptom and abnormal biochemical indicator;it may be used as an effective method for treating acute liver failure in pediatric patients.

X-连锁肾上腺脑白质营养不良(X-linked adrenoleukodystrophy,X-ALD)是一种最常见的过氧化物酶体病,呈X-连锁隐性遗传.在美国,男性发病率1/21 000、女性携带率1/14 000~([1]).日本男孩中,估计患病率在1/30 000～1/50 000,明显低于欧美国家~(2])。

党内民主是我们党的建设中一个重要的问题.党的十六大报告在讲到坚持和健全民主集中制问题时指出:"党内民主是党的生命,对人民民主具有重要的示范和带动作用."事实表明:没有党内民主,就没有党的兴旺发达.什么时候党内民主搞得比较活跃,我们党和党所领导的事业就充满生机和活力;什么时候党内民主受到削弱和破坏,党就会犯错误,党的事业就会遭受挫折.