一个常染色体显性遗传视网膜色素变性家系突变基因筛查

目的对一个四代常染色体显性遗传视网膜色素变性家系进行突变基因的筛查。方法选取了常见的11个视网膜色素变性（retinitis pigmentosa,RP）相关致病基因：视紫红质（rhodopsin,RHO,RP4）基因、视网膜变性慢蛋白（peripherin 2／retinal degeneration slow PRPH2／RDS,RP7）基因、视网膜色素变性1（retinitis pigmentosa 1,RP1）基因、前体mRNA处理因子31（pre—mRNA processing factor31,PRPF31,RP11）基因、次黄苷单磷酸脱氢酶1型（inosine monophosphate dehydrogenase 1,IMPDH1,RP10）基因、视杆外节蛋白1（rod outer segment protein 1,ROM1）基因、神经视网膜亮氨酸拉链（neural retina leucine zipper,NRL,RP27）基因、Pim1激酶相关蛋白1（piml—kinase associated protein1,PAP1,RP9）基因、视锥-视杆同源盒（cone—rod homeobox—containing,CRX）基因、前体mRNA处理因子3（pre—mRNA processing factor3,PRPF3,RP18）基因、前体mRNA处理因子8（pre—mRNA processing factor 8,PRPF8,RP13）基因。通过聚合酶链式反应扩增这11个RP相关致病基因的外显子和邻近拼接位点,扩增产物进行正、反双向测序。结果①对常见的11个RP相关致病基因的64个片段的研究显示,除了基因多态性和内含子的变异外,未发现与疾病有关的外显子或邻近拼接住点的突变。②首次在中国人种中发现RP9基因的c．629A→G,（Lys210Arg）变异。结论该文设计研究的突变基因未被检测到,表明此家系在11个RP相关致病基因的外显子和邻近拼接位点无基因突变。     

视网膜色素变性患者RP1基因点突变频率及其特征：101例分析

背景：视网膜色素变性(retinitis pigmentosa，RP)是一组最常见的遗传性致盲眼底病，在遗传和表型上均具有较大的异质性1999年Pierce等发现了一个新的视网膜感光细胞特异基因——RP1，之后的研究发现该基因的突变可导致常染色体显性遗传RP(autosomal dominant RP,adRP)。目前RP1的分子遗传学研究还主要集中在白种人群。目的：研究RP1基因在中国RP患者中的突变频率、特征及其在RP发病机制中所起的作用。设计：以RP患者为研究对象健康人为对照组的观察对比研究。单位：一所军医大学医院基因诊断治疗中心。对象：101例无亲缘关系的RP患者均系香港威尔士亲王医院眼科门诊及香港眼科医院1998—01／2001—12的就诊患者，年龄10～79岁(男43例，女58例)，平均年龄40岁。纳入标准：符合国内外RP诊断的通用标准者(包括眼底镜观察及视网膜电图测试)。排除标准：其他视网膜眼底病变患者。对照组为190例健康成年人(无RP家族史及眼部检查无RP及其它眼部疾患)，以确定所检测到的变异是否系RP1基因的多态现象。方法：在101例RP患者中运用构象敏感凝胶电泳(conformation sensitive gel electrophoresis,CSGE)和DNA直接测序方法检测RP1基因全编码区范围内的点突变。主要观察指标：RP1基因在中国RP患者中的突变频率、类型及在RP发病机制中的作用。结果：本研究中RP1基凶在所有RP患者中的突变检出率为1／101，突变最终导致RP1蛋白严重截短，疾病的表型可能源于功能性RP1蛋白合成量的不足。此外在本研究人群中还发现10个错义突变，除M4791的病理意义未确定之外，其余均系RP1基因的多态现象。结论：RP1蛋白中相应片段(密码子1052～1933)的缺失会导致RP的发生。大范围的RP1基因分型工作是有必要的．并且可同时发现更多的RP致病突变以及不同于其他种族人群的RP1基因多态变化，进而从根本上治疗RP，彻底提高其患者的生活质量。     

视网膜色素变性患者RP1基因点突变频率及其特征:101例分析

背景视网膜色素变性(retinitis pigmentosa,RP)是一组最常见的遗传性致盲眼底病,在遗传和表型上均具有较大的异质性.1999年Pierce等发现了一个新的视网膜感光细胞特异基因--RP1,之后的研究发现该基因的突变可导致常染色体显性遗传RP(autosomal dominant RP,adRP).目前RP1的分子遗传学研究还主要集中在白种人群.目的研究RP1基因在中国RP患者中的突变频率、特征及其在Rp发病机制中所起的作用.设计以RP患者为研究对象健康人为对照组的观察对比研究.单位一所军医大学医院基因诊断治疗中心.对象101例无亲缘关系的RP患者均系香港威尔士亲王医院眼科门诊及香港眼科医院1998-01/2001-12的就诊患者,年龄10～79岁(男43例,女58例),平均年龄40岁.纳入标准符合国内外RP诊断的通用标准者(包括眼底镜观察及视网膜电图测试).排除标准其他视网膜眼底病变患者.对照组为190例健康成年人(无RP家族史及眼部检查无RP及其它眼部疾患),以确定所检测到的变异是否系RP1基因的多态现象.方法在101例RP患者中运用构象敏感凝胶电泳(conformation sensitive gel electrophoresis,CSGE)和DNA直接测序方法检测RP1基因全编码区范围内的点突变.主要观察指标RP1基因在中国RP患者中的突变频率、类型及在RP发病机制中的作用.结果本研究中RP1基因在所有RP患者中的突变检出率为1/101,突变最终导致RP1蛋白严重截短,疾病的表型可能源于功能性RP1蛋白合成量的不足.此外在本研究人群中还发现10个错义突变,除M479I的病理意义未确定之外,其余均系RP1基因的多态现象.结论RP1蛋白中相应片段(密码子1052～1933)的缺失会导致RP的发生.大范围的RP1基因分型工作是有必要的,并且可同时发现更多的RP致病突变以及不同于其他种族人群的RP1基因多态变化,进而从根本上治疗RP,彻底提高其患者的生活质量.     

常染色体显性遗传视网膜色素变性家系的分子遗传学研究

目的对1例4代常染色体显性遗传视网膜色素变性家系进行致病基因定位,并对候选基因进行突变筛查。方法收集1例视网膜色素变性家系,抽取家系成员外周血并提取DNA,用连锁分析法对与疾病相关联的22个已知基因进行定位,并对定位区域内的候选基因进行突变筛查。结果两点连锁分析结果显示,在微卫星标记D7S484处取得最大LOD值为1.51（θ=0.00时）;聚合酶链反应直接测序法筛查候选基因RP9,未发现突变。结论该家系可能存在RP9基因大的碱基缺失突变或基因重排,也可能存在1个新的致病基因。     

基于Ion Torrent半导体测序技术的遗传代谢病基因诊断Panel临床应用

目的探讨基于Ion Torrent半导体测序的遗传代谢病基因诊断Panel的临床应用价值。方法收集109例串联质谱筛查可疑代谢病患儿及其父母的外周血标本,用基因Panel进行检测,可疑突变经Sanger测序验证。结果 109例患儿中有97例(88.99%)基因诊断明确,其中96例检测到存在与临床症状相符的2个等位基因致病突变,1例鸟氨酸氨甲酰转移酶缺乏症为男性半合子; 11例(10.09%)仅检测到1个等位基因致病突变; 1例(0.92%)未检测到致病性突变。结论基因Panel在临床疑似遗传代谢病的检测中有较高的阳性诊断率,可为串联质谱筛查后续临床治疗和遗传咨询提供依据。     

单核苷酸多态性与肿瘤转移的研究进展

单核苷酸多态性（SNP）是基因组变异最丰富的一种DNA序列变化形式，其与现有的遗传标记物结合，与现有的基因诊断体系接轨，将加速检验医学从表型诊断向基因型诊断的过渡。肿瘤是一种多基因遗传病，其侵袭转移与致病基因的SNP密切相关，现就近几年这方面的研究新进展作一综述。     

目标基因捕获测序技术检测1例肥厚型心肌病致病基因突变

目的通过靶向捕获高通量测序技术初步筛查肥厚型心肌病(HCM)致病基因突变。方法收集1例HCM患者临床信息和外周血,并提取基因组DNA,制备DNA全基因组文库。挑选导致HCM的8个候选基因,用GenCap基因序列捕获技术靶向富集该患者外周血DNA候选基因并行高通量测序。通过生物信息学分析筛选致病突变。用Sanger测序来验证相应致病基因突变位点。结果目标基因靶向捕获测序结果经与公共数据库和内部健康人测序数据库对比,发现致病基因突变位点MYBPC3 D770N,该致病基因突变与Sanger测序结果一致。结论用目标基因靶向捕获测序技术可实现对HCM致病基因突变的初步筛查,对HCM的临床基因诊断具有潜在的应用价值。     

Amp-FLP连锁分析法应用于Wilson病的产前基因诊断

为探索对Wilson病产前基因诊断的方法，应用13号染色体上3个STR位点的扩增片段长度多态性（Amp－FLP）为遗传标记，对3个WD家系进行产前诊断，结果2个胎儿为致病基因携带者，1个胎儿为患者。表明以这3个多态位点为遗传标记，能快速、简便、准确地对该病进行产前诊断。     

Amp—FLP连锁分析法应用于Wilson病的产前基因诊断

为探索对Ｗｉｌｓｏｎ病产前基因诊断的方法，应用１３号染色体上３个ＳＴＲ位点的扩增片段长度多态性（Ａｍｐ－ＦＬＰ）为遗传标记，对３个ＷＤ家系进行产前诊断，结果２个胎儿为致病基因携带者，１个胎儿患者，表明以这３个多态位点为遗传标记，能快速、简便、准确地对该病进行产前诊断。     

MLPA技术在DMD基因外显子拷贝数变异家庭基因诊断及产前诊断中的应用

目的分析多重连接依赖探针扩增(MLPA)技术在DXD基因外显子拷贝数变异家庭基因诊断及产前诊断中的应用价值。方法选取2018年1月至2019年1月就诊于该院产前诊断门诊,基因检测结果明确为DMD基因缺失或重复突变的14例杜氏肌营养不良症患儿(先证者)作为研究对象,通过使用MLPA技术针对DMD基因全部79个外显子进行缺失和重复突变检测,对先证者、先证者母亲及其再次妊娠所怀胎儿进行基因诊断及产前诊断。结果 DMD基因MLPA检测结果显示,先证者拷贝数缺失突变13例,重复突变1例;家系1～8先证者拷贝数缺失或重复片段遗传自其母亲;胎儿拷贝数杂合缺失突变3例,杂合重复突变1例,其余10例胎儿未检测到外显子缺失或重复。结论 MLPA技术是一种灵敏、准确、高效的分子诊断方法,该方法能对DMD基因外显子拷贝数进行相对定量,从而明确致病因素,更好地为杜氏肌营养不良症家庭优生优育提供指导意义。     

Gene mutations in retinitis pigmentosa and their clinical implications

Retinitis pigmentosa (RP) is a group of inherited progressive retinal diseases affecting about 1 in 3500 people worldwide. So far, there is no prevention or cure, with permanent visual loss or even blindness the ultimate consequence usually after midlife. The genetics of RP are complex. It can be sporadic, autosomal dominant, autosomal recessive, or X-linked. Thirty-two genes are known to be associated with RP, sometimes the same gene gets involved in different inheritance traits. Some RP cases have a digenic cause. About 60% RP cases still have no known genetic cause. A large number of mutations cause RP, and they can be deletions, insertions, or substitutions that cause missense mutations or truncations. The RHO, RP1, and RPGR genes contribute the greatest number of known mutations causative of RP. But there is no single mutation that alone accounts for more than 10% of unrelated patients. Genetic testing for RP therefore requires screening for a group of genes. High-throughput and automated sequence detection technologies are essential. Due to the complexity in phenotype and genetics, and the fact that RP is untreatable, genetic testing for presymptomatic diagnosis of RP is controversial. Meanwhile, new genes are still to be identified, mostly by family linkage and sib-pair analysis. Research on gene therapy for RP requires information on gene mutations causative of RP.     

网织血小板检测在血液病诊断与疗效评估中的应用

目的：探讨流式细胞仪检测网织血小板(Reticulated Platelet，RP)在血液病诊断与疗效评估中的应用。方法：运用双标记法。用血小板特异性抗体CD42-PE标记血小板，再用噻唑橙(Thiazole Orange,TO)染色RP内RNA，以健康人为对照，用流式细胞仪检测血液病患者在初诊、初治、缓解期的网织血小板水平。结果：血液病患者初诊病人RP％明显高于正常对照组，RP绝对值明显低于正常对照组：缓解病人RP％明显低于初诊病人，RP绝对值明显高于初诊病人，但与正常对照组均无显著性差异。结论：RP是对血液病患者进行临床诊断和疗效观察的一个重要指标。     

类风湿关节炎与复发性多软骨炎致环杓关节固定的诊断与治疗

目的 :探讨类风湿性关节炎 (RA)和复发性多软骨炎 (RP)的临床特点、诊断与鉴别诊断以及治疗与预后 ,以提高对RA和RP的认识。方法 :回顾性分析 11例RA和 15例RP患者的临床资料。结果 :1991～ 2 0 0 0年间收治 11例RA和 15例RP中 ,分别有 4例 ( 36 %)和 5例 ( 33%)累及双侧环杓关节。RP中 6 0 %有耳软骨炎 ,33%有鼻软骨炎 ,47%有呼吸系统受累 ,部分患者有心血管受累 ,用皮质激素、免疫抑制和氨苯砜控制症状 ,4例RA和 5例RP患者因呼吸道梗阻行气管切开术 ,3例RA患者行杓状软骨切除声带外展移位术。 15例RP中 1例治疗无效 ,死于呼吸道并发症。结论 :RA、RP是自身免疫性疾病 ,可累及多器官的软骨结构及结缔组织 ,在耳鼻喉科有表现 ,易误诊。早期诊断 ,采用综合治疗是提高疗效改善预后的关键。     

Activity index in rheumatic polymyalgias

OBJECTIVE: The activity of rheumatic polymyalgia (RP) was assessed using simplified activity index (RPAI). MATERIAL AND METHODS: 76 patients with a valid diagnosis of RP were treated in the Rheumatological Department of the Territorial Clinical Hospital #1; the clinical picture of the disease and its activity were evaluated according to RPAI, and the effect of glucocorticoids (GC) on the index was assessed. RESULTS: All the subjects had a high RPAI at admission (32.8 +/- 0.8). After three weeks of GC therapy RPAI decreased significantly. The frequency of RP recurrence correlated with the absence of remission after lowering prednisolone dose. CONCLUSION: Knowing the clinical manifestations and course of RP, the diagnosis is not difficult. However, for adequate treatment of patients the degree of disease activity needs to be assessed during monitoring, after dose lowering, and after prednisolone is discontinued.     

18F-FDG PET/CT诊断复发性多软骨炎

目的 探讨¹⁸F-FDG PET/CT对复发性多软骨炎（RP）的诊断价值。方法 回顾性分析28例RP患者（RP组）及28名健康志愿者（对照组）的¹⁸ F-FDG PET/CT表现,观察病灶部位、形态、边界及密度,计算病灶最大标准摄取值（SUV_max）。绘制ROC曲线,评价病灶SUV_max对RP的诊断效能。结果 28例RP患者中24例见气管及支气管密度增高、边界模糊及管壁增厚;4例见喉软骨密度增高、管壁水肿及增厚,边界模糊。共91个病灶,均位于鼻软骨、双侧耳廓软骨、肋软骨、喉软骨、气管及支气管软骨5个部位,各部位病灶SUV_max均高于对照组相同部位（P均<0.01）。ROC曲线结果示肋软骨、喉软骨、气管及主支气管软骨、双侧耳廓软骨SUV_max对RP的诊断价值均较高（AUC均>0.9）,鼻软骨诊断价值中等（AUC=0.71）。结论 RP患者¹⁸F-FDG PET/CT表现有一定特异性,可同时显示多部位病灶,有助于RP诊断。     

Molecular diagnostics for retinitis pigmentosa.

BACKGROUND: At least 1 million people worldwide have retinitis pigmentosa (RP), making it relatively common among the inherited forms of blindness. Mutations in many genes may cause RP. The most common known mutation, Pro347Leu in rhodopsin, is found in no more than about 1% of unrelated patients, implying the impracticality of a diagnostic test which would screen only for a few, common mutation sites. CONCLUSIONS: Ongoing discovery and study of RP genes makes it feasible to consider a molecular diagnostic test which would screen coding regions of all known RP genes by a mutation detection method such as conformation-sensitive gel electrophoresis followed by sequencing. The parallel development of RP genetic knowledge and treatments such as gene therapy will make such tests both possible and necessary.     

Composition of urinary glycosaminoglycans in a patient with relapsing polychondritis

OBJECTIVES: Several investigators have reported an increase in urinary glycosaminoglycans (GAGs) in patients with relapsing polychondritis (RP). The aim of this investigation is to analyze the composition and structure of urinary GAGs from a Brazilian patient with RP. DESIGN AND METHODS: The identification and structural analyses of the GAGs were made by electrophoresis and degradation with specific enzymes and identification of their disaccharides products by HPLC chromatography. RESULTS: The disaccharide products formed from RP urinary chondroitin sulfate (CS) by action of chondroitin ABC lyase showed a substantial relative increase of nonsulfated disaccharides with a relative decrease of 6-sulfated disaccharides compared to control subjects. In addition, a significant change of the ratio of CS and heparan sulfate was also observed in the RP patient. CONCLUSION: The RP patient analyzed has shown a structural anomaly of the urinary CS and this may contribute to the diagnosis of this disease.     

Adult‐onset Diamond‐Blackfan anemia with a novel mutation in the exon 5 of RPL11: too late and too rare

Diamond‐Blackfan anemia (DBA) is a congenital erythroid aplasia usually diagnosed in the early infancy and associated with mutations or large deletions in 11 ribosomal protein (RP) genes. Adult patients with severe, transfusion dependence, aregenerative anemia might have a genetic‐in‐origin disease with an atypical presentation. Late onset nonclassical DBA should be ruled out and mutations of RP genes studied.     

Use of diagnostic clusters to assess the economic consequences of rhinopharyngitis in children in Italy and France during the winter. Rhinitis Survey Group

To determine how practitioners diagnose rhinopharyngitis (RP), we conducted a longitudinal, multicenter study of a cohort of 900 children, collecting medical and economic data without interfering with usual medical practice during the winter of 1996-1997 in France and Italy. All ear, nose, and throat (ENT) infections were described clinically; data on the consumption of medical items (physician visits, drug treatment, hospitalization, physiotherapy, preventive treatment, laboratory tests, roentgenograms, and outpatient procedures) were collected to estimate the cost of caring for patients with RP. The mean age of the children was 28.0 months, and the ratio of males to females was approximately 5 to 4. Patients had had a mean 4.1 episodes of RP the previous year and 1.4 episodes of acute otitis media (AOM). There were no marked differences in the children's characteristics between France and Italy. During the winter of the study, this population experienced 4.26 episodes of ENT infection, of which 73.5% were documented at the study sites. Seven homogeneous groups of RP were found, 2 of them each representing <4% of the overall population. One group presented with otalgia, although the diagnosis of AOM was not recorded by the physician. In 4 groups, the presence of nasal discharge plus cough (without otalgia) was used to make the diagnosis. Medical item consumption varied by country and by group of RP, mainly in the prevailing choice of antibiotics. The difference in duration of treatment was not statistically significant. As a consequence, the costs of caring for patients with RP varied greatly, RP with AOM being the most costly. Last, prognostic factors for costly episodes of infectious ENT were identified. The population at risk included young children who had had AOM episodes during the previous winter, had a first episode of AOM before 6 months of age, had a history of AOM associated with effusion, or attended a community-based child care facility. Therefore, clinical trials aimed at demonstrating cost-effectiveness of prophylaxis should focus on this population.