高迁移率族蛋白B1的临床研究进展

高迁移率族蛋白B1(high mobility group protein box 1,HMGB1)是一组高度保守的DNA结合蛋白,存在于体内各种细胞的细胞核中,调控着细胞基因的转录.生理状态下HMGB1 发挥着核结合蛋白的作用.当释放到细胞间隙后则会起到晚期炎症因子的作用.最近的研究提示,HMGB1 不仅参与脓毒症、自身免疫性疾病、慢性肝病、恶性肿瘤等的致病过程,还可参与细胞损伤的修复或致病,在多种疾病及组织器官损伤、修复过程中发挥着重要作用.     

新生鼠缺氧缺血性脑损伤Fas相关磷酸酯酶和氧化还原因子-1mRNA的表达

围产期缺氧缺血性脑病 (HIE)是严重危害新生儿健康和生命的疾病。由于氧化还原因子 1 (Ref 1 )在DNA碱基切除修复通路中起重要作用[1 ] ;Fas相关磷酸酯酶 (Fas associatedphosphatase 1 ,FAP 1 )通过负性调节影响Fas介导的凋亡[2 ] ,本研究利用新生鼠缺氧缺血性脑病损伤 (HIBD)动物模型 ,采用原位杂交方法观察Ref 1及FAP 1在脑中的表达情况。材料和方法1 .实验动物及模型的制备 :7日龄新生Wistar大鼠 (重庆医科大学动物中心提供 ) ,体重 1 2～ 1 6g。按文献方法甲氧氟烷吸入麻醉后 ,结扎左侧颈总动脉。恢复 2h ,通入含氧体积浓度为…     

蛋白质精氨酸甲基转移酶在血液肿瘤疾病中的作用研究进展

蛋白质精氨酸甲基转移酶(protein arginine methyltransferases,PRMTs)家族具有广泛的分子功能,在干细胞发育系统中参与造血细胞的发生、发育、增殖和分化。在血液肿瘤疾病中,PRMTs可通过甲基化途径参与许多重要的生物学过程,包括细胞增殖、细胞周期控制、基因转录、DNA损伤修复。PRMTs的异常表达可导致肿瘤的发生与发展,特别是在儿童白血病中,通过靶向PRMTs治疗,降低PRMTs的表达,可以有效地抑制肿瘤细胞的增殖和存活。因此,PRMTs与血液肿瘤疾病的关系备受关注,未来很可能成为治疗的一个重要靶标。该文对PRMTs在血液肿瘤疾病中的作用及其机制作一综述,以期为血液肿瘤疾病的治疗提供新的思路。     

小儿急性白血病P16基因突变的分析

分析小儿急性白血病 P1 6基因的突变及其在疾病发生的作用。利用 PCR扩增 P1 6基因的外显子 1、外显子 2 ,并进行缺失突变分析 ,利用均一变性凝胶电泳( CDGE)对外显子 2进行点突变分析。结果显示 :小儿急性白血病缺失突变率异常高 ,5 7例患者中有 1 8例发生缺失突变 ,只有 6例发生点突变 ,总突变率为 31 .6 %。ALL患者 P1 6基因的总突变率明显高于 AML患者 ,为 6 4 .7% ,而 AML患者 P1 6基因的总突变率只有 8.7%。我们认为 :P1 6基因的突变在小儿急性白血病发生中起重要作用。P1 6基因含有“CG”DNA重复顺序 ,是小儿急性白血病 P1 6基因缺失突变率高于点突变率的原因。     

伴DNA修复相关基因胚系突变的遗传易感性小儿白血病5例临床分析

目的探讨DNA修复相关基因胚系突变所致遗传易感性小儿白血病的诊断思路以及这类疾病临床、遗传学特征。方法收集并回顾性分析2017年5月—2020年1月确诊的5例DNA修复相关基因胚系突变的遗传易感性小儿白血病患儿的临床资料及遗传学、分子学资料。结果伴遗传易感性的白血病患儿中涉及DNA修复缺陷者5例,男2例,女3例。5例中1例TP53突变,2例PMS2突变,1例9p21.3p21.1缺失,1例CHEK2突变。结论白血病诊疗中应注意体貌异常及肿瘤家族史,不存在以上也不能除外肿瘤遗传易感性。需甄别结构性变异,重视基因突变及杂合性缺失两种形式,完全缓解后的拷贝数变异检测应更加积极。TP53c. 421T C p. C141R的胚系突变形式为致病性突变。PMS2的单等位基因突变也可导致典型的结构性错配修复综合征发生。DNA修复缺陷相关胚系突变本身不影响化疗反应,体细胞改变特征才是影响化疗疗效的关键因素。     

小儿先天畸形重点实验室小儿横纹肌肉瘤p16基因缺失及突变的研究

目的探讨小儿横纹肌肉瘤p16基因缺失及突变的临床意义.方法应用控制模板DNA量的银染PCR-SSCP技术检测18例小儿横纹肌肉瘤手术切除标本中p16基因第1外显子.结果18例中有7例发生纯合性缺失,其纯合缺失率为38.8%(7/18);1例发生突变,缺失,突变率为44.3%.结论p16基因的纯合性缺失是p16基因失活的主要机制,在小儿横纹肌肉瘤的发生和发展中起重要作用.     

药物相关性分子靶标检测在指导Ⅲ-Ⅳ期儿童神经母细胞瘤治疗中的初步经验

目的 探讨药物相关性分子靶标检测在儿童神经母细胞瘤个体化治疗中的作用和意义.方法 2011年4月至2013年4月间在上海交通大学医学院附属新华医院诊治的13例儿童神经母细胞瘤患儿,取其手术肿瘤及外周血标本共31份(手术/二次手术标本共15份,外周血标本16份).通过免疫组化,酶联免疫吸附试验,荧光原位杂交,PCR扩增测序等方法检测常见肿瘤药物的相关分子靶标.共检测12种常用化疗药物和1种靶向药物的相关性基因的表达或突变.结果 共检测肿瘤标本分子靶点7个,包括:DNA拓扑异构酶ⅡA(TOPOⅡA),微管蛋白β3(Tubulinβ3),人切除修复交叉互补基因(ERRC 1),DNA拓扑异构酶Ⅰ(TOPO Ⅰ),DNA修复蛋白-O6甲基鸟嘌呤-DNA-甲基转移酶(MGMT),二氢叶酸还原酶(DHFR),胸苷酸合成酶(TS);外周血标本分子靶点2个,包括:CYP2C9*3和二氢叶酸还原酶DHFR(C829T)基因多态性测序.根据靶标对应的相关药物进行总结分析,研究结果提示:氟尿嘧啶类,长春碱类,环磷酰胺和甲氨蝶呤四类化疗药物敏感性较高;而紫杉醇类、拓扑替康/伊立替康、替莫唑胺/卡莫斯汀/司莫斯汀、蒽环类/依托泊苷和铂类的药物敏感性较低.研究共检测肿瘤标本和外周血标本涉及的靶向药物基因2个:血管内皮生长因子受体(VEGFR-2)和细胞间粘附分子1(ICAM-1).结果提示:贝伐单抗敏感性高.依据上述检测结果,我们及时调整治疗策略,优化化疗方案,取得了良好的效果,并为部分患儿创造了手术机会.结论 本研究在儿童神经母细胞瘤的多学科治疗过程中引入药物相关性分子靶标的检测,对于开展规范化治疗基础上的个体化治疗方案的选择,提供了依据和经验.     

PCR-CTPP技术在先天性巨结肠SIP1基因多态性研究中的应用

目的 研究用相对的两对引物一聚合酶链反应(polyrnerase chain reactionwith confronting two-pair primer,PCR-CTPP)技术在SIPl基因多态性检测中的应用,探讨先天性巨结肠症(Hirschsprung disease,HSCR)SIP1基因单核苷酸多态性特点.方法 采用PCR-CTPP技术检测分析60例HSCR患儿SIPl基因第8外显子多态性,并与聚合酶链反应一限制性片段长度多态性(PCR-Restriction fragment length polymorphism,PCR-RFLP)技术检测结果对比,探讨PCR CTPP在单核苷酸多态性(SNP)检测及分析中的应用.结果 经PCR-CTPP扩增出SIPl基因第8外显子CTG→CCG与GGC→GAC三种基因型相应的DNA片段,与DNA测序结果符合.HSCR符合杂合性突变.结论 PCR-CTPP技术成本较低、省时、操作简便,结果稳定可靠,对于SNP研究具有较高的应用价值.HSCR患儿SIPl基因单核苷酸多态性值得深入研究.     

表观遗传学在过敏性疾病发生中作用的研究进展

<正>过敏性疾病已成为全球性的临床和公共健康问题,严重危害儿童的身心健康。虽然遗传在过敏性疾病发生中起着重要作用,然而,基因并不能完全解释近年来过敏性疾病发病率快速增加的现象。已有较多研究表明,环境因素可以通过引起机体表观遗传学变化,从而导致过敏性疾病的发生。表观遗传学由Waddington~[1]首先提出,即DNA序列不变,但其基因表达发生可遗传的变化,机制涉及DNA甲基化、组蛋白修饰及非编码RNA等。     

小儿横纹肌肉瘤p16基因缺失及突变的研究

目的;探讨小儿横纹肌肉瘤P16基因缺失及突变的临床意义,方法：应用控制模板DNA量的银染PCR－SSCP技术检测18例小儿横纹肌肉瘤手术切除标本中P16基因第1外子,结果,18例中有7例发生纯合性缺失,其纯合缺失率为38．8％（7／18）,1例发生突变,缺失,突变率为44．3％,结论：P16基因的纯合性缺失是P16基因失活的主要机制,在小儿横纹肌肉瘤的发生和发展中起重要作用。     

DNA repair: Pathways and defects

Damaged DNA can be repaired by three different mechanisms: photoreactivation, excision repair and postreplication repair. Each mechanism is regulated by a highly specific set of enzymes. Defects within these systems result in diseases which have one common feature: affected individuals are cancer prone. Recently, newly developed methods not only make it possible to diagnose affected patients but also to detect individuals at risk. Furthermore, the results obtained elucidate some mechanisms of carcinogenesis. Clinical applications are discussed.     

Gene therapy for disorders affecting children, progress and potential

For over two decades gene therapy has been actively pursued as a treatment modality for the inherited diseases that affect the paediatric population, however, it is still to make a real impact in the clinic. There are many reasons for this including inadequate technology and a lack of understanding of the biological complexities that impact on the efficiency of gene delivery and its outcomes, both positive and negative. However, recent progress is now addressing these issues and indicates that these problems can be overcome, and that gene therapy will play a significant role in the treatment of at least some of these disorders. This review will first give a short overview of relevant gene delivery technologies, what strategies can be used and which diseases are potential targets for gene therapy, and then illustrate several specific diseases for which gene therapy is actively being developed.     

Midkine, A New Heparin-Binding Growth/Differentiation Factor: Expression and Distribution during Embryogenesis and Pathological Status

Midkine (MK) is a 13 kDa heparin-binding growth factor found as a product of a retinoic acid-responsive gene. MK is rich in basic amino acids and cysteine and its sequence is not homologous with any other proteins so far reported, so it is a new family of heparin-binding growth factor. It has been found that MK exerts variety of biological activities such as neurite-promoting, neuronal cell survival and differentiation-inducing activities. MK is strictly expressed during the mouse embryogenesis; among the adult organs, it is detected only in the kidney. MK is also strongly expressed in a number of human carcinomas and specifically localized in senile plaques in the brain of patients with Arzheimer disease. More recently, it has been reported that MK is an important molecule regulating inflammation response and tissue repair. These results demonstrated that the relevance of MK not only in normal development, but also in processes leading to tissue repair or diseases. Increased MK gene expression is a common phenomenon observed in many human carcinomas, therefore MK is of significant interest in cancer biology. As a new growth/differentiation factor, many issues including the detailed sites and the precise time of MK expression, the exact cellular source which synthesizes and secretes MK, the signal transducing receptors for MK, the mechanisms underlying those developmentally regulated expression and its potential clinical significance still remain unknown. To elucidate the molecular mechanisms of MK action will lead not only to a deeper understanding of developmental processes, but also to the ultimate obtaining a key to diagnose and treat human carcinomas.     

间充质干细胞向上皮细胞的诱导分化及应用研究进展

间充质干细胞( mesenchymal stem cell,MSC)在体内外可被诱导分化为多种上皮细胞,如肺泡上皮、肾小管上皮和角膜上皮细胞等,表达上皮细胞特异性标志.MSC回输体内后,可有效修复损伤的肺上皮细胞组织,缓解炎症损伤,减轻肾小管细胞损伤、减少肾间质炎症细胞浸润,促进肾小管上皮细胞再生,并抑制细胞凋亡;也可...     

间充质干细胞治疗难治性肺疾病研究进展

难治性肺疾病是严重威胁患者健康的呼吸系统疾病,可导致肺功能受损甚至呼吸衰竭,目前常规的药物治疗效果欠佳,因此,寻找更有效的治疗方案尤为重要。间充质干细胞(MSCs)是一类多功能干细胞,可通过调节炎症和免疫反应,促进受损细胞修复以及通过旁分泌促进受损组织的再生,有望为难治性肺疾病的治疗提供新的治疗手段。     

天然免疫缺陷病的研究进展

天然免疫缺陷是原发性免疫缺陷病的一种.根据最新文献报道,天然免疫缺陷病主要包括七种疾病,这些疾病发病的根本原因均为基因突变.其中三种疾病的病因与NF-кB信号通路受阻有关,而有两种疾病与一般原发性免疫缺陷病发病机制不同,患者被相关的病毒感染后才会诱导发病.天然免疫缺陷在原发性免疫缺陷病中占有较重要的位置,但是由于对此病的认识不足,临床上很多天然免疫缺陷的疾病无法确诊.     

Balloon dilatation in children for oesophageal strictures other than those due to primary repair of oesophageal atresia,interposition or restrictive fundoplication

Background Balloon dilatation of the oesophagus in children has been performed predominantly for treating strictures, which are the result of primary repair of oesophageal atresia, interposition surgery or restrictive Nissen's fundoplication. Reports of the use of this technique for alternative causes of stricture are few.Objective To report our experience and success with balloon dilatation of strictures due to caustic ingestion, achalasia, oesophagitis, congenital stenosis, and epidermolysis bullosa (EB) and to make comparisons with our treatment of patients with primary repair of oesophageal atresia (OA), as well as with reports in the English language literature.Materials and methods Retrospective review of fluoroscopically guided balloon oesophageal dilatation procedures in 19 patients over a 5-year period, and comparison of those performed for OA repair complications with those due to other diseases. The average radiation dose, per procedure, was calculated by a medical physicist.Results Ten patients had strictures as a result of primary repair of OA. Three patients had stricture as a result of EB, two from achalasia, two from caustic injury, one due to an oesophageal web and one from reflux oesophagitis. Our results show that the technique can also be curative for the last group and that it may be used intermittently to alleviate symptoms in ongoing diseases. We have not experienced any complications and have also calculated that, even with prolonged use of multiple procedures, the radiation exposure is comparable to other radiological techniques.Conclusions Patients with alternative causes for oesophageal stricture may be treated to resolution within 2 years using balloon dilatation. Ongoing diseases such as EB require ongoing dilatation, but balloon dilatation of strictures has been successful in alleviating swallowing difficulty. Patients with stricture from OA repair sometimes need ongoing dilatation. Radiation exposure for multiple procedures, over an extended period, is comparable to that from a single abdominal CT, and can be considered acceptable when repeat complex surgery is the alternative treatment option, or when ongoing incurable disease is the cause of the stricture.This paper was presented in a modified form at the 39th Annual Congress of ESPR, Bergen, Norway, June 2002     

Update on the immunodeficiency diseases

Newer understanding of the development of the immune system and novel methods to repair these defects promise cures for previously fatal diseases. The advances stem from the ability to define the developmental stages and describe the diseases at the gene level. Coordinate with this progress, recombinant DNA technology may soon permit the precise correction of the defect. For the moment, gene replacement is accomplished at the cellular level by bone marrow transplantation. Here too, rapid clinical advances now permit the use of nonsibling donors. The long-term results are equivalent to those obtained in the past when only HLA-matched siblings could donate. There is now a support group that is of inestimable value to the families of those who suffer from immunodeficiency. Thus, there has been both technological and social progress in the area of primary immunodeficiency. The future is clearly brighter each year.     

全外显子测序技术在危重症新生儿遗传病中的应用价值

目的 探讨全外显子组测序（WES）技术在危重新生儿遗传病中的应用价值。方法 选取于该院新生儿重症监护室治疗的66例疑似遗传病或诊断不明的危重新生儿为研究对象。收集患儿临床资料,采集患儿及其父母静脉血行WES检测,完成遗传病因诊断。结合患儿的临床表现寻找相关的致病基因变异。结果 66例疑似遗传病或诊断不明的危重新生儿中,男34例,女32例,其中通过WES检测出有基因变异14例（21%）;1例患儿经WES检测未见有基因变异,但因临床表现高度怀疑为色素失禁症,联合多重连接酶探针依赖扩增技术,检测到IKBKG基因4~10号外显子的杂合缺失变异。15例检测出基因变异的患儿中,致病性基因变异10例（67%）;可疑致病性基因变异1例（7%）;基因变异意义未明4例（27%）。15例患儿中有13例行染色体检查,只有1例染色体异常。结论 染色体检查不能作为遗传病的确诊手段,WES检测技术是寻找疑似或诊断不明的危重新生儿遗传病的重要工具,然而WES技术有一定的局限性,可联合其他测序方法进行检测。