婴儿期发病的腓骨肌萎缩症家系PMP22基因突变研究

目的：研究一个婴儿期发病的腓骨肌萎缩症（CMT）家系PMP22基因突变,探讨该家系CMT的遗传学特点。方法：应用STR结合多重PCR法对2例CMT患儿及该家系内15名表型正常的成员进行PMP22基因重复突变的分析。同时选择20名健康人做为对照。结果：在2名CMT患儿及5名家系内表型正常的成员中发现了PMP22基因重复突变,其中5例突变在STR位点D17S921,2例突变在STR位点D17S4A,而家系内其余10名成员及20名健康人未发现突变。结论：该CMT家系的致病基因为17p11.2-p12区域内包含PMP22基因的重复突变,其亚型为CMT1A。     

SCN1A 基因变化在家族性热性惊厥发病中的作用

目的探讨SCN1A基因变化在家族性热性惊厥发病中的作用。方法收集8个家族性热性惊厥家系的临床资料,留取先证者和部分家系成员的血液标本,PCR扩增所有先证者SCN1A基因编码的26个外显子及其上下游侧翼序列至少50个以上碱基,并测序。对新发现的碱基变化,再对家系其他成员进行相应外显子的基因序列筛查,从而明确变异起源。同时纳入200例同年龄正常对照者加以验证。结果 8例先证者存在33种碱基变化,其中32种被提交为单核苷酸多态性,一处为新发现的错义突变。家系4先证者第15外显子存在一错义突变(C.2650GA),该碱基变化尚未见报告,且证实突变遗传自先证者父亲。200名正常对照者相应外显子测序未发现该碱基变化。C.2650GA导致SCN1A第884位的甘氨酸为丝氨酸所替代(Gly884Ser)。突变位于电压门控钠离子通道α亚基第2同源结构域的第4次跨膜和第5次跨膜之间(DIIS4-S5),通过蛋白序列比对,该氨基酸高度保守。结论 SCN1A基因突变与家族性热性惊厥的发病有关,电压门控钠离子通道α亚基电压感受器(S4)及离子通道孔(S5-S6)外区突变可能是引起表型相对较轻的癫痫综合征的重要原因。     

先天性Q-T间期延长综合征3型一家系基因突变分析

目的 对一个表现有先天性Q-T间期延长综合征,同时表现有扩张性心肌病、心脏传导障碍性疾病的家系进行相关基因分析,探讨其发病机制.方法 一个中国人3代多表现型的先天性Q-T间期延长综合征家系,采集8名家系成员和100名健康对照血样,采用PCR扩增、DNA直接测序技术进行SCN5A、KCNQ1、KCNH2、LAMIN A/C基因突变分析,采用多聚酶链反应--单链构象多态性进行基因型和表型研究.结果 PCR-DNA直接测序在先证者钠通道SCN5A基因第26号外显子发现了9个碱基CAGAAGCCC缺失突变,该突变位于SCN5A基因DⅢ和DⅣ连接区域,导致第1507位谷氨酰胺、第1508位赖氨酸、第1509位脯氨酸3个氨基酸缺失.家系中患者检出与先证者相同的突变,而家系内参加本研究的健康人和家系外100名健康对照未出现这种突变.结论 SCN5A基因突变是部分先天性Q-T间期延长综合征的分子发病机制,SCN5A基因delQKP1507-1509突变是国内未曾报道过的新突变,国外的相同突变报道仅表现为Q-T间期延长综合征,无其他表型.进一步开展SCN5A delQKP1507-1509突变的功能研究有助于理解相关疾病的分子发病机制.     

癫痫患儿5例基因型表型分析及临床研究

目的　分析5例癫痫患儿的临床资料,探究SCN9A基因突变相关性脑病的临床特点。方法　选取2017年9月至2018 年12月中国医科大学附属盛京医院小儿神经内科就诊的80例癫痫患儿,完善二代测序检测存在SCN9A基因突变且SCN9A基因考虑为致病相关基因,对测序结果进行致病性分析,观察并记录患儿临床表现。结果　5例患儿均存在SCN9A基因突变,且均为错义突变,患儿存在局灶性发作,全面强直阵挛发作及肌阵挛发作,部分患儿存在智力或运动发育落后。结论　SCN9A基因突变多为杂合突变,本研究中发作形式为局灶性发作较多,可与发热相关,临床表型多样。     

癫痫患儿5例基因型表型分析及临床研究

目的　分析5例癫痫患儿的临床资料，探究SCN9A基因突变相关性脑病的临床特点。方法　选取2017年9月至2018 年12月中国医科大学附属盛京医院小儿神经内科就诊的80例癫痫患儿，完善二代测序检测存在SCN9A基因突变且SCN9A基因考虑为致病相关基因，对测序结果进行致病性分析，观察并记录患儿临床表现。结果　5例患儿均存在SCN9A基因突变，且均为错义突变，患儿存在局灶性发作，全面强直阵挛发作及肌阵挛发作，部分患儿存在智力或运动发育落后。结论　SCN9A基因突变多为杂合突变，本研究中发作形式为局灶性发作较多，可与发热相关，临床表型多样。     

Dravet综合征少见致病基因突变及临床表型分析北大核心CSCD

目的研究Dravet综合征（Ds）少见致病基因突变,并对携带少见致病基因突变的患儿进行基因型-表型：分析。方法收集2005年2月至2016年8月在北京大学第一医院儿科就诊的Ds患儿,采用Sanger测序及多重连接依赖的探针扩增（MLPA）方法筛查SCN1A及PCDH19基因突变,对SCN1A及PCDH19基因突变阴性的患儿进行癫痫基因靶向捕获二代测序;分析携带少见致病基因突变患儿的临床表型特点。结果在670例Ds患儿中,SCN1A突变阳性者556例（83．0％）,PCDH19突变阳性者6例,余108例SCN1A及PCDH19突变筛查阴性者送检癫痫基因靶向捕获二代测序,发现12例携带其他致病基因突变,其中GABRA1杂合突变3例,GABRG2杂合突变2例,GABRB2杂合突变2例,SCN2A杂合突变1例,TBC1lD24复合杂合突变2例,ALDH7A,复合杂合突变2例。6例PCDH19杂合突变者发作均有丛集性,仅1例病翟甲曾出现癫痫持续状态;2例GABRB2杂合突变者发作控制相对好;2例TBC1lD24复合杂合突变者均出现多次肌阵挛持续状态;2例ALDH7A1复合杂合突变者加用维生素B6发作控制。未发现GABRG2、SCN2A和GABRA1基因突变导致的Ds表型有特异性。结论中国Ds患儿可出现的少见致病基因突变包括PCDH19、GABRG2、SCN2A、GABRA1、GABRB2、TBC11D24和ALDH7A1基因。GABRB2和TBC11D24基因突变的发现扩展了Ds的致病基因谱。PCDH19突变者丛集性发作突出,癫痫持续状态少见;GABRB2突变者发作相对容易控制;TBC1lD24复合杂合突变者肌阵挛发作持续状态突出。     

SCN1A基因突变致遗传性异卵双生Dravet综合征家系分析并文献复习

目的总结遗传性异卵双生Dravet综合征家系临床及SCN1A基因突变特点。方法分析华中科技大学同济医学院附属同济医院2017年6月收治的异卵双生Dravet综合征患儿兄妹各1例和全身性热性惊厥附加症(GEFS+)母亲家系的临床特征,进行SCN1A基因测序。并结合文献分析基因突变类型与Dravet综合征的关系。结果 2例患儿及其母亲均携带未报道过的SCN1A基因突变c.3624AT(p.R1208S),该突变位于蛋白的罕见区域(Na+通道α亚基第2个环)。SCN1A基因突变以点突变多见,约占93.8%,突变位置与Dravet综合征临床表型的相关性复杂。结论该病例为国内首次报道的遗传自双亲之一的异卵双生Dravet综合征。SCN1A基因突变c.3624AT(p.R1208S)发生于蛋白罕见区域,目前国际尚未报道。     

全面性癫(癎)伴热性惊厥附加症的临床表型和SCN1A基因筛查研究

目的 分析全面性癫(癎)伴热性惊厥附加症(generalized epilepsies with febrile seizures plus,GEFS+)的临床特点,并对部分患者进行SCNIA基因筛查,寻找基因突变.方法 收集两个GEFS+家系的临床资料,并进行分析;留取先证者和部分家系成员的血液标本,通过变性高效液相色谱法(denaturing hish performance liquid chromatography,DHPLC)等方法进行SCNIA基因筛查、测序及序列分析.结果 (1)两个家系共101名成员,其中受累者28例(男、女各14例).发作表型有热性惊厥(FS)7例、热性惊厥附加症(FS+)6例、FS+伴失神发作1例、FS+伴肌阵挛发作1例.未发现严重发作表型.另外,有肯定的临床发作,但由于不能获得详细的临床资料而不能进行发作分类者13例;两个家系都符合常染色体显性遗传,其中一个家系存在双系遗传现象.(2)GEFS+家系B的先证者和家系正常对照均发现SCN1A第9外显子存在A>G突变(c.1212A>G),系一多态性位点;SCN1A其余外显子未发现突变.结论 本研究在GEFS+家系B仅发现G/A多态现象,未发现SCN1A致病性突变,支持其遗传异质性;病因学有待进一步研究.     

婴儿期发病Lesch-Nyhan综合征4例临床表现及HPRT1基因突变分析并文献复习

目的　分析婴儿期发病Lesch-Nyhan综合征（LNS）患者临床特点以及相关基因突变情况。方法　对2017—2018年就诊于首都儿科研究所附属儿童医院的4例尿酸升高、运动发育迟缓、肌张力障碍伴或不伴有自残行为的婴儿患者进行全外显子基因检测,生物信息学分析筛选致病性变异,并对核心家系成员采用Sanger测序分析突变来源。总结整理Pubmed、知网和万方数据库报道过的婴儿期发病LNS中国患者的临床特点及基因突变情况。结果　4例均携带HPRT1基因致病性突变,符合LNS遗传诊断。突变分别为2个错义突变（c.49T＞A,p.Y17N和c.133A＞G,p.R45G）,1个无义突变（c.325C＞T,p.Q109*）和1个移码突变（c.419delG,p.G140Afs*26）,其中p.Y17N和p.G140Afs*26为未报道的新突变。回顾文献收集 11例婴儿期发病的中国LNS患者,HPRT1基因型与临床表型的相关性分析发现了智力迟缓、运动发育迟缓、高尿酸血症为核心表型,且出现比例为100%;和错义突变相比,截短性突变较早出现自我伤害表型。结论　该研究描述了婴儿期发病的中国LNS患者临床与基因信息,补充了HPRT1基因突变谱,对临床表现为高尿酸血症合并神经系统异常的儿童患者进行LNS基因筛查将有助于早期LNS诊断和治疗。     

先天性肌强直两家系的临床分析及CLCN1基因突变检测

目的研究先天性肌强直(MC)两家系的临床特点及CLCN1基因突变情况。方法对两个MC家系7例患者的临床资料进行分析。从两个家系成员及100例正常对照者外周静脉血中提取基因组DNA,通过PCR及DNA测序对CLCN1编码区进行突变分析。结果两个家系的7例患者均为幼年起病的肌强直与运动笨拙,伴有肌肥大和扣击性肌球,肌电图示肌强直电位发放。基因检测发现先证者1及家系1其他患者在第8外显子892位核苷酸处发生G-A杂合突变,使298位丙氨酸被苏氨酸代替(A298T)。而另一家系患者及其他健康成员CLCN1基因23对外显子直接测序均未发现有突变存在。结论在中国MC家系中发现了新的CLCN1突变位点,为研究中国人群CLCN1突变类型、遗传方式提供了一定的参考。     

Bibliometric data: a disaster for many non-American biomedical journals

Bibliometric data published by the Institute of Scientific Information in Philadelphia (ISI), and which was previously discussed in Acta Paediatrica , has increasingly been used despite all the relevant and severe criticism that has been raised against this method of evaluating individual research results and grading scientific journals. It is obvious that the present trend regarding the use of bibliometric data as a basis for priorities and funding of research and for the promotion of individual scientists favours American-oriented research projects at the expense of those that are based on concepts of predominantly European relevance.

Conclusion: For the future of non-American research, it is important that no single super-power, i.e. the USA, should dominate scientific priorities. The condition for efficient European competition is that European Centres with high levels of competence for creative research and training of scientists from all over the world are established. In addition, it is important that the results of European research are published in prestigious European journals, as was the situation before World War II.     

The importance of different immunosuppressive regimens in the development of posttransplant diabetes mellitus

Solid-organ transplantation is the optimal long-term treatment for most patients with end-stage organ failure. After solid-organ transplantation, short-term graft survival significantly improved (1). However, due to chronic allograft nephropathy and death with functioning graft, long-term survival has not prolonged remarkably (2). Posttransplant immunosuppressive medications consist of one of the calcineurin inhibitors in combination with mycophenolate mofetil (MMF) or azathioprine (Aza) and steroids. All of them have different adverse effects, among which posttransplant diabetes mellitus (PTDM) is an independent risk factor for cardiovascular (CV) events and infections causing the death of many transplant patients and it may directly contribute to graft failure (3). According to the criteria of the American Diabetes Association (4), diabetes mellitus (DM) is defined by symptoms of diabetes (polyuria and polydipsia and weight loss) plus casual plasma glucose concentration ≥ 11.1 mmol/L or fasting plasma glucose (FPG) ≥ 7.0 mmol/L or 2-h plasma glucose level ≥ 11.1 mmol/L following oral glucose tolerance test (OGTT). This metabolic disorder occurring as a complication of organ transplantation has been recognized for many years. PTDM, which is a combination of decreased insulin secretion and increased insulin resistance, develops in 4.9/15.9% of liver transplant patients, in 4.7/11.5% of kidney recipients, and in 15/17.5% of heart and lung transplants [cyclosporine A (CyA)/tacrolimus (Tac)-based regimen, respectively] (5). Risk factors of PTDM can be divided into non-modifiable and modifiable ones (6), among which the most prominent is the immunosuppressive therapy being responsible for 74% of PTDM development (7). Emphasizing the importance of the PTDM, numerous studies have determined the long-term outcome. On the basis of these studies, graft and patient survival is tendentiously (8) or significantly (9, 10) decreased for those developing PTDM.     

alpha -SMOOTH MUSCLE ACTIN DISTRIBUTION IN THE PULMONARY VASCULATURE COMPARING HYPOPLASTIC AND NORMAL FETAL LUNGS

We investigated the intra-acinar pulmonary vascular muscularization in the developing human fetal lung between the 17th and 24th gestational weeks, that is, during the canalicular phase of lung development. Fifteen hypoplastic and 25 normal developed lungs were included in this study using monoclonal alpha -smooth muscle (sm) actin antibodies for smooth muscle detection. Computer-aided image analysis was performed for morphometrical measurements and statistical evaluation. Alphasm-actin-immunoreactive intra-acinar vessels down to a luminal diameter of less than 10 mu m were detected in hypoplastic as well as in normally developed lungs. Crucial differences presented as follows: significantly higher density of intra-acinar vessels, especially due to alpha -sm-actin-negative vessels less than 30 mu m in luminal diameter, in the control group; significantly higher alpha -sm-actin immunoreactivity per section unit as well as per vessel in the hypoplastic lung group. As suggested by others, alpha-sm-actin-positive cells of the intra-acinar vessel wall in the developing human lung were demonstrated to be smooth muscle cells, their immediate precursors, and pericytes. We conclude that the increased alpha -sm-actin immunoreactivity represents muscularization of the vessel wall in functional terms and may be regarded as one structural cause among others for the establishment of persistent fetal circulation in hypoplastic lungs.     

Whole-body mineral measurements in Swedish adolescents at 17 years compared to 15 years of age

Aim: To provide reference data for bone mineral variables in 15- and 17-y-old adolescents and to analyse the relationships between these variables and measures of bone and body size, gender, puberty, growth, various lifestyle and environmental factors and socioeconomic background.

Methods: In the same 321 randomly selected adolescents (147 boys and 174 girls) living in two different regions of Sweden, the total bone mineral content (TBMC), bone area (BA) and total bone mineral density (TBMD) were assessed by dual-energy X-ray absorptiometry at ages 15 and 17 y. The effects of bone and body size, gender, growth, sexual maturity, physical activity, region of domicile, social conditions, food habits, smoking and alcohol intake on TBMC and TBMD were examined in multivariate analyses.

Results: In the 15-y-old adolescents, BA, height, gender, physical activity, maturity and weight explained 91% and 48%, of the variance in TBMC and TBMD, respectively. In similar analyses in the 17-y-olds, the corresponding figures were 92% and 62%, respectively, when BA, height, growth, physical activity, gender and region emerged as significant in the model. In all these analyses, BA explained most of the variance in TBMC and TBMD. No significant reduction of variance was found when different measures of social conditions, smoking, food habits, alcohol or dietary intakes of energy, calcium or vitamin D were included in the models. The reason why region of domicile had a significant impact on TBMC in the 17-y-olds is not known. The fact that the normal fluoride concentration in drinking water (1.1 mg/L) is 10 times higher in the region where TBMC was higher than in the other region is an interesting observation.

Conclusion: Almost 90% of the variance in TBMC and 50% of that in TBMD was explained by measures of bone and body size and only a few percent by gender, physical activity, Tanner stage, growth and region of domicile.     

VARICELLA ZOSTER VIRUS INFECTIONS IN THE PEDIATRIC STEM CELL TRANSPLANT PATIENT

Varicella zoster virus (VZV), a member of the human herpesvirus family, causes the clinical syndromes of chickenpox during primary infection and shingles on later reactivation. In immunocompromised patients, including those undergoing hematopoietic stem cell transplantation, VZV can produce life-threatening infections. The most serious forms of VZV infection involve hematogenous dissemination of the virus to vital organs, such as the lung, brain, and liver. Advances in immunoprophylaxis, antiviral chemotherapy, and vaccine development have provided effective tools to limit the morbidity and mortality previously associated with VZV infection in hematopoietic stem cell transplant patients. In this review, we discuss virologic aspects of VZV, pathogenesis of VZV infection, methods of viral diagnosis, clinical manifestations of infection in both normal and immunocompromised patients, and available preventative and therapeutic measures.     

The potential role of rapamycin in pediatric transplantation as observed from adult studies

Although pediatric patient and renal graft survival rates have shown marked improvements during the past decade, the persistent toxicities of immunosuppressive drugs and chronic allograft attrition remain major obstacles in transplant therapy. Results in adult patients suggest that complete steroid withdrawal is possible in the majority of recipients under treatment with a cyclosporin A-rapamycin (CsA RAPA) regimen. Furthermore, preliminary studies suggest that a marked reduction in the dose of CsA may be possible under the umbrella of RAPA coverage. The gain in immunosuppressive efficacy afforded by RAPA has not only been obtained without an increased morbidity owing to infectious or neoplastic causes, but also with the potential for reducing the incidence and/or progression of chronic rejection.     

GENETICS OF CARNEY COMPLEX AND RELATED FAMILIAL LENTIGINOSES, AND OTHER MULTIPLE TUMOR SYNDROMES

Carney complex is a multiple endocrine neoplasia (MEN) syndrome that affects the adrenal cortex, the pituitary and thyroid glands, and the gonads. The complex is also associated with skin and mucosa pigmentation abnormalities and myxoid and other neoplasms of mesenchymal and neural crest origin. Thus, this syndrome also belongs to another group of genetic disorders, the lentiginoses (or lentigenoses), which include the Peutz-Jeghers, LEOPARD, arterial dissections and lentiginosis, and Laugier-Hunziker syndromes, Cowden disease and Ruvalcaba-Myhre-Smith (Bannayan-Zonana) syndrome and the centrofacial, benign patterned and segmental lentiginoses, all of which can be associated with a variety of developmental defects. The inheritance of Carney complex, just like that of the other MENs and the lentiginoses, is autosomal dominant. Genetic loci or genes have been identified for Carney complex, Peutz-Jeghers and Ruvalcaba-Myhre-Smith syndromes, but not for other lentiginoses. Elucidation of the molecular defects responsible for these disorders is expected to shed light on aspects of early neural crest differentiation, the regulation of pigmentation, the development of autonomous endocrine function, and endocrine and nonendocrine tumorigenesis.     

All you ever wanted to know about infant formulas (but were afraid to ask)

The World Health organisation recommends breast feeding infants for the first six months of life. When this breast feeding does not occur either through parental choice or medical need, infant formulas will be required. There is a bewildering array of formulas on the UK market for many different requirements. When faced with an unsettled infant many parents (and health care professionals) will experiment with the infant formula available and often attend the paediatric clinic looking for help and advice. It is therefore essential that paediatricians understand what milks are available and what the key differences between different products are. This review attempts to provide a simple guide through many of the formulations currently available in the UK; and offers advice for the dietary management of the child with extra calorie requirements, infants with cow's milk protein allergy, gastro oesophageal reflux disease, apparent unresolved hunger and infantile colic. Whatever the underlying condition, there is likely to be an infant formula that is suitable in this generation of ever expanding formulations.     

Génétique des schizophrénies : mise en perspective des schizophrénies à début précoce et autres pathologies du développement

Schizophrenia (SCZ) is a severe brain disorder characterized by hallucinations, delusions, flat and/or inappropriate affect and cognitive impairment. The lifetime risk is about 0.5% with heritability of 65–85%. The prevalence of early-onset schizophrenia (defined here as before 15 years of age) has not been well studied, but is likely to be 5–10% of all cases. The rarity of early-onset SCZ has made it difficult to study. We focus on genetic studies of adults with schizophrenia, highlighting results for early-onset schizophrenia where available. Prior to the past 5 years, studies failed to find replicable association or linkage between SCZ and specific genes when appropriate statistical corrections for multiple testing were used. Many false positive results were probably reported using the candidate gene approach. Recently, the development of single nucleotide polymorphism (SNP) “chips” has permitted large genome-wide association study (GWAS) analyses that suggest that across all age groups, a proportion of genetic risk can be attributed to a large number of common SNP, each with a very small effect on risk (odds ratios of 1.1 or less). The greatest known genetic effect is conferred by the 1.5–3 Mb 22q.11.2 deletions, which occurs in ∼ 1/4000–1/6000 births with SCZ developing in 20–30% of carriers. Large SNP and aCGH microarray studies have now identified associations between SCZ and other rare, large copy number variations (CNV, insertions and deletions) with high odds ratios (5–10), including deletions of 1q21, 2p16.3 (neurexin-1 gene), 3q29 and 15q13.3, and duplications of 16p11.2. Some of these CNV are also associated with autism or other developmental disorders as well as epilepsy or intellectual deficiency, suggesting some overlap in the mechanisms that contribute to risks of these disorders. Based on preliminary data from larger-scale analyses in progress, approximately 1–2% of cases carry a CNV that has been clearly associated with SCZ (ORs 4–12). Whole exome and genome sequencing studies of large adult samples will be the next steps to identify rarer SCZ-associated mutations, including point mutations and smaller as well as rarer CNV. Genetic findings are beginning to contribute to an understanding of biological mechanisms of SCZ risk and may lead to new approaches to treatment.     

INACTIVATION OF PULMONARY SURFACTANT AND THE TREATMENT OF ACUTE LUNG INJURIES

Inactivation of pulmonary surfactant may be important in acute lung injury and acute respiratory distress syndrome. Treatment of surfactant dysfunction by instilling exogenous surfactants may improve gas exchange and pulmonary mechanics. Surfactants used for treatment vary in their attributes and effects, so when various surfactants are considered for therapy, resistance to inactivation is an important consideration. Animal models of acute lung injury exist in which the relative merits of surfactants can be compared. We hypothesize that the surfactants most resistant to inactivation in vitro will be the ones that are most effective in treatment of animal models of acute lung injury. Surfactants with higher concentrations of surfactant proteins (specifically A, B, and C) are more resistant to inactivation. Nonionic polymers mimic surfactant proteins in preventing surfactant inactivation under some conditions. Adding nonionic polymers to surfactant containing minimal amounts of SP-B and SP-C markedly improves lung function of animals with lung injury. Making surfactants more "inactivation-proof" may improve surfactant therapy of acute lung injuries.