毛囊角化病患者ATP2A2基因新剪切位点突变的检测研究

目的:检测1例严重表型毛囊角化病患者ATP2A2基因的突变.方法:采用聚合酶链反应及直接测序法对该患者进行ATP2A2基因的突变位点检测,运用反转录技术检测突变导致的RNA水平的变化,同时对120名无血缘关系健康者作为对照进行测序验证.结果:通过筛查在患者ATP2A2基因中发现一个新的剪切位点突变IVS18+5G＞C,反转录分析证实该突变的发生导致ATP2A2基因转录后在其突变等位基因的18号和19号外显子之间插入了27个核苷酸.结论:该例毛囊角化病患者的检测结果进一步扩充了ATP2A2基因的突变库,并为阐明ATP2A2基因的突变导致该病发生的分子机制提供了帮助.     

毛囊角化病ATP2A2基因突变分析

目的检测毛囊角化病患者ATP2A2基因的突变。方法提取全部患者及健康对照个体的外周血DNA,采用聚合酶链反应扩增ATP2A2基因的全部外显子,并进行DNA测序。结果在收集到的2个家系和3例散发患者中共发现3个突变,包括1个缺失突变(1622delAACA),1个插入突变(180insCTTAA)和1个错义突变(698GT),均为未见报道的突变。在100例正常对照中均未发现上述突变。结论收集到的毛囊角化病患者存在ATP2A2基因的突变,这些突变可能会影响角质形成细胞中钙离子的转运,使表皮细胞的连接和分化出现异常。     

一毛囊角化病家系ATP2A2基因突变检测

目的:检测一毛囊角化病家系的ATP2A2基因突变。方法:提取2例患者外周血DNA,采用聚合酶链式反应及DNA直接测序方法,检测患者ATP2A2基因突变。结果:该家系患者及其两女儿存在ATP2A2基因的碱基缺失突变,即ATP2A2基因第10个外显子1220位开始缺失了AA 2个碱基。而该家系中其他正常者未发现此突变。结论:ATP2A2基因第10个外显子1220delAA突变可能与该家系患者临床表型有关。     

毛囊角化病ATP2A2基因突变的检测

目的 :检测1个家系及1例散发毛囊角化病患者ATP2A2基因突变情况。方法 :提取家系中2例患者、5例正常人及1例散发患者的外周血DNA,采用聚合酶链反应(PCR)对其ATP2A2基因进行扩增,并对其扩增产物进行测序,检测该基因突变点。对照组为100例无血缘关系的健康人。结果:发现1个新的错义突变(R603I)和1个已报道的错义突变(R131Q)。结论:该研究发现毛囊角化病患者存在ATP2A2基因的突变,这些突变可能影响肌浆/内质网钙ATP酶(SERCA)2的结构和功能,使表皮细胞的连接和分化出现异常,最终导致疾病的发生。     

哈萨克族毛囊角化病一家系ATP2A2基因突变研究

目的 探讨哈萨克族毛囊角化病一家系患者ATP2A2基因突变.方法 收集哈萨克族毛囊角化病49人家系的临床资料,采集44名家系成员和100例无亲缘关系健康人外周血,提取基因组DNA.采用PCR和DNA测序对该家系进行ATP2A2基因突变检测.结果 该家系毛囊角化病遗传方式属于常染色体显性遗传.家系中11例患者在ATP2A2基因12号外显子的剪切位点发生杂合突变(1288-1G→A),即第1288-1位碱基由G突变为A,而家系中33例正常成员及100例健康对照均未发现该突变.结论 该家系毛囊角化病发病可能是由ATP2A2基因12号外显子的剪切位点发生杂合突变(1288-1G→A)所致.     

毛囊角化病一家系中ATP2A2基因新的剪接突变

目的: 检测毛囊角化病一家系中ATP2A2基因新的剪接突变。方法: 提取家系中2例患者和2名正常人外周血DNA,采取聚合酶链反应技术对ATP2A2基因进行扩增,并对其产物进行测序,以100名正常人作对照。结果:该家系中患者ATP2A2基因的第13号内含子第1761+2位碱基由胸腺嘧啶(T)转化为胞嘧啶(C)。结论: 该家系发病可能是由ATP2A2基因发生剪接突变所致。     

毛囊角化病一家系ATP2A2基因突变的检测

目的检测一毛囊角化病家系的ATP2A2基因突变。方法提取患者外周血DNA,采用聚合酶链反应(PCR)及DNA直接测序方法,检测患者ATP2A2基因突变。结果该家系先证者及其母亲、舅舅存在一个新的ATP2A2基因错义突变,即在ATP2A2基因第18外显子核苷酸2728位碱基A变成G(c.2728 A→G),导致患者在910位(p.910N→D)将天冬酰胺(AAC)替换为天冬氨酸(GAC),而该家系中其他正常者未发现此突变。结论研究结果支持单倍型遗传是毛囊角化病显性遗传的一种常见机制。     

毛囊角化病九例ATP2A2基因突变研究

目的：检测9例毛囊角化病(Darier's disease, DD)患者的基因突变。方法：提取1家系中2例患者、7例散发患者及100名正常对照外周血基因组DNA,用Sanger测序检测9例DD患者ATP2A2的致病突变。对不携带ATP2A2突变的患者,应用全外显子组测序(WES)寻找可能导致该疾病的其他变异。364名健康对照被纳入突变筛选。可疑的变异通过Sanger测序进行确认。结果：通过Sanger测序,我们发现9例DD患者中有5例出现了ATP2A2突变,包括1例报道过的突变和3例新突变。在其余病例中,WES鉴定了四个ATP酶基因(ITPR3、ATP2B2、RYR1和PLCB1)中的4个变异,均参与钙信号通路。结论：本研究中发现了4个ATP2A2基因突变,其中3个为新发突变,丰富了ATP2A2基因突变谱,并提示四种ATP酶基因变异可能与DD相关。     

毛囊角化病一家系及三例散发患者ATP2A2基因突变研究

目的：对毛囊角化病（Darier's disease, DD）一家系及三例散发患者进行ATP2A2基因的突变分析。方法：收集先证者及其家系成员、散发病例的临床资料和外周血,采用PCR技术扩增ATP2A2基因所有编码区及侧翼序列,用Sanger法测序检测潜在的突变,选取与患者无亲缘关系的100例健康人作为对照,同时对已报道的ATP2A2基因突变进行文献回顾。结果：家系中三例患者均检测出ATP2A2基因第5号外显子c.380 G>A(p.G127D)新发错义突变;散发患者S1检测出第13号外显子C.1676G>A(p.R559Q)错义突变,散发患者S2检测出第14号外显子c. 2001C>T(p.D667D)同义突变,散发患者S3未检测出突变。结论：本研究中共发现三个突变,其中c.380G>A(p.G127D)在中国人群中首次报道,拓展了ATP2A2的基因突变谱。     

角化性皮肤病

20110564毛囊角化病一家系中ATP2A2基因新的剪接突变/孙忠辉(温州医学院),李明,张国龙∥中国麻风皮肤病杂志.-2010,26(11).-762~764为了检测毛囊角化病一家系中ATP2A2基因新的剪接突变,提取家系中2例患者和2名正常人外周血DNA,采用PCR对ATP2A2基因进行扩增,并对其产物进行测序,同时设立正常对照。结果发现患者ATP2A2基因的第13号内含子第1761+2位碱基由T转化为C。认为该家系发病可能是由ATP2A2基因发生剪接突变所致。图3参8(赵恩兵)20     

Darier disease--novel mutations in ATP2A2 and genotype-phenotype correlation

Darier disease (DD) is with a frequency of up to 1 in 36,000 a relatively common genodermatosis with autosomal dominant inheritance and late age of onset. The progressive skin manifestations are variable, but often debilitating and disfiguring, and may be associated with a wide range of neuropsychiatric problems, such as epilepsy and depression. On histology, acantholysis and dyskeratosis are prominent findings, implicating impaired functionality of desmosomes. Recently, mutations in the ATP2A2 gene encoding SERCA2, a calcium pump of the endo/sacrcoplasmic reticulum, have been identified as the molecular basis of DD. This slow-twitched calcium ATPase has two splice variants, one of which is highly expressed in epidermis, and maintains low intracellular calcium levels by facilitating transport of cytosolic calcium into the endoplasmic reticulum. Thus, it may confer a direct effect on the established calcium-dependent assembly of desmosomes. We screened ATP2A2 in a cohort of 24 DD families using conformation sensitive gel electrophoresis and direct sequencing, and detected 14 distinct mutations, 9 of which were novel. The mutational spectrum included 9 missense mutations, 1 nonsense mutation, 3 small in-frame deletions, and a 19-basepair insertion. Mutations were scattered over the entire gene with a slight preponderance in the first 8 exons, and affected exclusively residues conserved among all SERCAs. In addition, we found 2 silent polymorphisms, 1 of which occurred in 4 unrelated families. Comparison of molecular data and phenotypic features, such as severity and type of disease, occurrence of mucosal involvement, or association with neuropsychiatric disorders, did not reveal an obvious genotype-phenotype correlation in our cohort.     

Transcutaneous PO2 and PCO2 measurements in various skin lesions

Low tcPO2 levels were observed in various skin lesions, indicating that most pathological changes in the skin induce such a reduction. In contrast, the tcPCO2 changed less, with the exception of marked elevations in bullous or prenecrotic lesions. This procedure may have practical applications in clinical dermatology, particularly in quantitating the response to therapy and in predicting a necrotic change.     

Interleukin-2 receptor expression and interleukin-2 production in bullous pemphigoid

Summary Expression of interleukin-2 (IL2) receptors was studied on peripheral blood lymphocytes (PBLs) in 25 patients with bullous pemphigoid. Analysis was carried out by flow cytometry. Without immunosuppressive therapy expression of IL2 receptors only on T cells (CD5) was significantly increased as shown by double staining (11.9%±7.8% vs 2.1%±1% in controls). After 2–3 weeks under immunosuppressive treatment with prednisolone and azathioprine, however, BP lymphocytes did not exhibit any IL2 receptors. PBLs of 18 BP patients showed an increase in IL2 production (1027.4±670.5 U/ml vs 270±100 U/ml in controls) during the acute stage of the disease after stimulation with phythaemagglutinin (PHA)/ phorbol myristate acetate (PMA). On the contrary, IL2 production of the same cells in five patients was only in the lowest range of control values after PHA stimulation without PMA (12.5±29.6 U/ml vs 40±20 U/ml in controls). Under treatment with immunosuppressants the IL2 production normalized after PHA/PMA stimulation and slightly decreased following PHA stimulation. From these results we conclude that a T-cell activation via activation antigens, as IL2 receptors, and the production of the specific ligand, IL2, may play a role in the pathogenesis of bullous pemphigoid, especially in the earliest stages, and serve as a marker of disease activity.     

Herpes zoster in 2 sisters

Transmission of herpes zoster infection from one sister to the other is described, resultant from close everyday contacts. Clinical manifestations of the disease (severity, dissemination, course and type of involvement) were much more marked in the elder sister, suffering from disseminated Darier's dyskeratosis and marked debility. Herpes was complicated with vasculitis, necrosis, Pseudomonas aeruginosa infection, development of pneumonia and keratitis. Problems of treatment of such patients are discussed.     

芹黄素对过氧化氢所致黑素细胞凋亡的作用

目的采用过氧化氢(H2O2)诱导的黑素细胞体外氧化应激模型探讨芹黄素对黑素细胞的抗氧化保护作用。方法采用MTT法测定不同浓度芹黄素预处理前后H2O2对正常人表皮黑素细胞活力的影响,An-nexin-V/PI双染流式细胞术检测细胞的凋亡情况,DCFH-DA流式细胞术检测细胞活性氧(ROS)的含量。结果 250μmol/L的H2O2作用于黑素细胞24h,黑素细胞活力降低至(36.42±7.21)%,凋亡率增加至(48.82±12.55)%。0.6μmol/L,2.5μmol/L和10.0μmol/L芹黄素预处理1h后,H2O2所致细胞活力分别增加至(41.53±5.25)%,(45.33±6.28)%和(51.92±5.29)%,凋亡率减少至(42.25±7.69)%,(37.54±8.82)%和(32.25±58.28)%。250μmol/L的H2O2作用于黑素细胞2h后,黑素细胞ROS含量增加至空白对照组的(69.22±8.57)倍。10μmol/L芹黄素预处理1h后H2O2所致黑素细胞ROS含量为空白对照组的(54.48±5.03)倍,低于单用H2O2处理组。结论芹黄素可能通过减少ROS生成和抑制氧化应激对HO所致黑素细胞凋亡具有保护作用,其可能具有开发为治疗白癜风新药的前景。