一个先天性特发性眼球震颤家系中致病基因FRMD7的突变研究

目的 研究一个先天性特发性眼球震颤家系的致病基因.方法 选取X染色体上微卫星标记物,通过PCR扩增后,进行基因组扫描.应用GeneMapper软件进行PCR扩增产物片段大小和单倍型分析,Linkage 5.1软件进行两点法连锁值(Log of odds,LOD)计算,通过基因序列分析发现致病基因突变.结果 经两点法计算,在DXS1047可获最大LOD值为8.55;基因序列分析发现FRMD7基因第9外显子存在G990T的杂合性基因突变.结论 FRMD7基因突变是导致该家系出现疾病的主要原因.     

先天性眼球震颤遗传学研究进展

先天性眼球震颤是一种不伴有其他明显眼病的眼科遗传病,该疾病的发生有很高的遗传异质性,已经发现多个致病基因基因座,Xq26–q27、Xp11.4-p11.3、13q31-q33和6p12,可能还有新的基因座没有被发现,已经研究的基因座致病基因侯选区域很大,在该区域的侯选基因太多,所以必须精细定位后,才有可能完成致病基因的克隆工作。本文对先天性眼球震颤遗传学的研究进展进行了综述。     

先天性特发眼球震颤一家系

先证者(Ⅴ10)男,18岁。生后2个月发现患儿眼球左右摆动。右眼视力为0.08,左眼视力为0.10,双眼矫正视力均为-10.0D→0.2。双眼呈水平性眼球震颤,无中间带,无快慢相,向各方向转动无变化。伴有弱视、近视和斜视(外斜约35°)。精神紧张时眼球震颤频率加快,遮盖一只眼检查视力时摆动频率增加更为显著;精神放松时,摆动频率明显降低。无代偿头位。前后房、虹膜色素分布正常。眼底:双眼视盘边界清楚,血量走行正常,黄斑部无明显色素,中心反射未见。视网膜正常,晶状体透明,眼角膜薄,无法手术治疗,诊断为先天性特发眼球震颤。家系调查:该家系6代共44人…     

先天性小眼球致病基因的研究进展

先天性小眼球是一种先天发育异常性眼科疾病,遗传方式有常染色体显性遗传,常染色体隐性遗传和X连锁隐性遗传。迄今为止,用连锁分析和细胞遗传学方法对小眼球相关基因进行了基因定位并进一步对候选基因进行突变分析。本文就近年来先天性小眼球致病基因研究方面作一综述。     

震颤分析对帕金森病、特发性震颤和增强的生理性震颤的鉴别诊断价值

目的 探讨震颤分析在帕金森病、特发性震颤和增强的生理性震颤中的鉴别诊断价值,以期为震颤的分类诊断提供客观的判断标准。方法 选取福建省立医院神经内科2020年12月至2021年12月收治的以震颤为主要表现的帕金森病患者15例（PD组）,及同期收治的16例特发性震颤患者（ET组）和17例增强的生理性震颤患者（EPT组）为研究对象。用肌电图仪进行震颤测定,分析在静息、姿势、负重500 g、负重1000 g下的震颤峰频率、收缩模式。结果 在PD、ET和EPT组中,静息、姿势和负重500 g和1000 g时的震颤频率分别约为4～6.5 Hz、5～8 Hz和8～10Hz。PD组、ET组和EPT组在静息、姿势和负荷后的震颤频率差异均有统计学意义（P<0.05）。PD组同步肌电图交替性收缩13例,同步性收缩2例,ET组和EPT组均为同步性收缩。结论 在PD、ET和EPT患者中,震颤频率和肌肉收缩模式是震颤分析中有价值的判断指标。     

A2M基因多态性与帕金森病和特发性震颤的关联

目的探讨α2-巨球蛋白基因（α2-macmglobulin gene，A2M）第24外显子1000G／A及第18外显子5’端剪接点5个碱基插入／缺失（insertion／deletion，IZD）多态与中国北方帕金森病（Parkinson’sdisease，PD）和特发性震颤（essential tremor，ET）的关联。方法应用聚合酶链反应-限制性片段长度多态性方法，对87例PD、73例ET和100名健康对照者A2MG／A和I／D两个位点的基因型和等位基因分布频率进行检测。结果（1）A2M基因G／A位点基因型和等位基因分布，在PD与ET、对照组间差异有统计学意义（P〈0．05），PD组的G等位基因和GA基因型明显高于ET、对照组（P〈0．05）；而ET与对照组间相比，差异无统计学意义（P〉0．05）。（2）A2M基因IZD位点基因型和等位基因分布，在PD、ET、对照组间差异无统计学意义（P〉0．05）。结论（1）G／A位点多态与PD的发病有关联，G／A位点多态与ET无关。（2）I/D位点多态与中国北方PD、ET无关。     

特发性震颤与帕金森病患者伴发抑郁症状的比较

目的:研究特发性震颤与帕金森病患者伴发抑郁症状的异同,以及特发性震颤患者伴发抑郁症状的发生率及特征.方法:对32例特发性震颤与122例帕金森病患者应用汉密尔顿抑郁量表(Hamilton Depression Scale,HAMD)进行评分,根据评分对2组患者伴发抑郁症状的发生率及抑郁严重程度进行评定,并比较2组患者抑郁量表评分、伴发抑郁症状的发生率及在HAMD量表单个项目中病例分布的差异.结果:特发性震颤组患者HAMD平均评分为(10.1±7.2)分,帕金森病组患者HAMD平均评分为(10.5±6.8)分,(P>0.05);特发性震颤患者伴发抑郁症状的发生率为43.8%,帕金森病患者伴抑郁症状的发生率为48.4%,(P>0.05);两组患者在HAMD量表各单个项目中病例分布均相似(P>0.05).结论:特发性震颤和帕金森病患者伴发抑郁症状不仅发生率接近而且临床表现也类似,特发性震颤患者伴发抑郁症状较常见,临床医生应提高认识.     

中国东北汉族一个先天性白内障家系致病基因的鉴定

目的鉴定一个先天性白内障家系的致病基因。方法根据已知与先天性白内障有关的12个致病基因的染色体上的定位,分别选取3～4个的微卫星标记位点,对该家系进行连锁分析。通过测序鉴定致病基因。结果在1q21.1GJA8位点显示最大Lod值2.44。致病基因定位于1q21.1区的GJA8基因,构成缝隙连接的缝隙连接蛋白Connexin50。DNA序列分析鉴定显示其第2外显子的第191个碱基杂合突变T>G导致其蛋白产物第64位缬氨酸转变为甘氨酸。结论Connexin50的V64G新生突变是导致该家系的致病原因。     

三个先天性遗传性白内障家系的致病基因突变分析CSCD

目的明确3个有家族史的先天性白内障家系的致病基因及突变类型,为家系的遗传咨询及产前诊断提供依据。 方法应用外显子组结合目标区域捕获测序芯片对先证者进行突变基因及突变位点捕获,Sanger测序对家系成员进行验证。结果家系1为多形性白内障,家系2为蓝点状白内障,家系3为珊瑚状白内障。3个家系的遗传方式均符合常染色体显性遗传。家系1患者均检测出CRYβB2基因c.463C〉T（p.Q155X）无义突变,家系2患者均检测出CRYGD基因c.43C〉T（ p．R14C）错义突变,家系3患者均检测出CRYGD基因c.70C〉A（p.P23T）错义突变;这些突变均表现为基因型与疾病共分离。结论家系1、2和3的致病性突变分别为CRYβB2基因c.463C〉T（p.Q155X）突变、CRYGD基因c.43C〉T（p.R14C）突变和CRYGD基因c.70C〉A（p.P23T）突变,本研究结果为家系的遗传咨询及产前诊断提供了依据.     

一个先天性小眼畸形家系的全外显子组测序分析及产前诊断

目的分析1个先天性小眼畸形家系的临床表型及遗传学病因。方法应用高通量测序技术对先证者及其父母进行全外显子组测序,筛选候选致病位点,对其家系进行Sanger测序验证,并通过羊水穿刺和Sanger测序为先证者母亲提供产前诊断。结果全外显子组测序和Sanger测序发现家系中的3例患者均携带OTX2基因c.289C>T(p.R97*)杂合变异,先证者母亲亦携带该变异,但无小眼畸形。先证者的父亲、舅母和胎儿未携带上述变异。结论OTX2基因c.289C>T(p.R97*)杂合变异很可能是该家系的发病原因。上述诊断将有助于该家系的遗传咨询和产前诊断。     

Congenital Nystagmus and Sleep: A Replication

In 8 subjects with congenital nystagmus, clearly manifested nystagmus was present while awake, with eyes open and closed, and during a variety of experimental tests, but could not be definitely discerned during REM and NREM sleep stages. However, conjugate REMs were present during the REMPs of all 8 subjects and could not be distinguished from patterns recognized in normal subjects. Brief episodes of reduced amplitude series of jerks were occasionally observed which could be considered to be “depressed” nystagmus, but such phenomena have been observed in the records of normal subjects. This replicates the findings of our previous report.     

Identification of a novel GPR143 mutation in a large Chinese family with congenital nystagmus as the most prominent and consistent manifestation

Congenital nystagmus is characterized by involuntary, rhythmical, repeated oscillations of one or both eyes. We studied a large Chinese family with nystagmus as a prominent and consistent manifestation phenotype in nine patients to map and identify a disease-causing gene for nystagmus. X-linked recessive inheritance was observed in the family, and foveal hypoplasia was detected in some of the nine patients. The disease gene was mapped to an approximately 10.6 Mb region flanked by DXS996 and DXS7593 on Xp22 with a significant peak multipoint LOD score. Analysis of 21 candidate genes in the region revealed a novel p.S89F mutation in the second transmembrane domain of GPR143, a G protein-coupled receptor which causes ocular albinism when mutated. All male patients in the family were hemizygous for the mutation; the female carriers were heterozygous for the mutation. The p.S89F mutation was not identified in 100 normal females or 100 normal males. Our results indicate that a mutation in the GPR143 gene can cause a variant form of ocular albinism, with congenital nystagmus as the most prominent and only consistent finding in all patients in this Chinese family. These results expand the spectrum of clinical phenotypes associated with GPR143 mutations. J.Y. Liu and X. Ren contributed equally to this work.     

A Case of Congenital Central Hypoventilation Syndrome with PHOX2B Gene Mutation in a Korean Neonate

Congenital central hypoventilation syndrome (CCHS) is a life-threatening disorder with apnea and cyanosis during sleep requiring immediate endotracheal intubation during the first day of life. The PHOX2B gene has been identified as the major gene involved in CCHS. This is the first report of a Korean neonate with CCHS confirmed to have a PHOX2B mutation with expanded alleles containing 20 polyalanine repeats that is a relatively small number compared to previous cases. The patient required intermittent ventilator support during sleep only and did not suffer from any other disorders of the autonomic nerve system. He consistently needs ventilator support during sleep and remains alive. Analysis of PHOX2B gene is useful for diagnosis and appropriate therapeutic intervention of CCHS patients.     

GFP和RFP报道基因在人体不同细胞中转导率的比较研究

目的 比较慢病毒载体介导的绿色荧光蛋白（GFP）和红色荧光蛋白（RFP）报道基因在人体不同细胞中的转导率,为再生医学和组织工程学研究中报道基因的选择提供依据.方法 将构建的GFP和RFP慢病毒载体分别转染人骨髓间充质干细胞（hMSC）、人脐静脉内皮细胞株（Eahy926）和人肺泡上皮细胞株（A549）.4d后,用流式细胞仪检测GFP和RFP的转导率,并利用激光共聚焦显微镜观察GFP和RFP的表达特征.结果 GFP和RFP在hMSC、Eahy926和A549三种细胞间的转导率均有显著性差异（P﹤0.01）;在hMSC或Eahy926或A549的同种细胞中GFP和RFP的转导率也有显著性差异（P﹤0.01）;RFP在部分Eahy926和A549细胞局部高表达.结论 慢病毒载体介导的GFP和RFP报道基因在人体不同细胞中的转导率明显不同.     

包裹E1A基因的纳米粒子制备及转染人肺腺癌细胞A549实验研究

目的制备包裹E1A基因（腺病毒早期表达基因）的纳米粒子,并观察其介导E1A基因转染人肺腺癌细胞A549的可行性和效率。方法应用聚乳酸聚乙醇酸共聚物和聚乙烯醇包载E1A基因,制备纳米级粒子混合物,检测其包埋率、体外释放情况及粒径大小。用制备的包裹DNA纳米粒子转染人肺腺癌细胞A549,并以阳离子脂质体为对照,用PCR、RT-PCR方法分别检测转染细胞中E1A基因DNA整合和mRNA表达。结果制备的纳米粒子粒径为150～280nm,包埋率为0.78%,体外释放约为22d;在转染相等质量的DNA情况下,纳米组所得克隆数较脂质体组多（P〈0.05）;PCR、RT-PCR结果表明纳米粒子和脂质体转染细胞均有E1A基因整合和mRNA表达。结论成功制备了纳米粒子,纳米粒子可携带外源基因进行基因转染。     

北京市甲型H1N1型流感病毒基因检测与流行趋势分析

目的 通过对甲H1N1型流感病毒基因的检测分析,揭示其基因变异情况及探讨甲型流感病毒的流行趋势,为甲型流感的防控提供理论依据.方法 样本为患者鼻咽部的粘膜上皮细胞和分泌物.取200μL标本置于核酸分离纯化系统中提取核酸,然后以LightCycler 480 PCR仪进行核酸扩增,最后分析曲线判断结果.每份标本均对甲型H1N1流感病毒基质蛋白基因（M基因）、核蛋白基因（NP基因）、血凝素基因（HA基因）、人类的RNA酶P基因（RNase P基因）进行检测和分析.结果判定：M基因、NP基因、HA基因、RNase P基因均阳性,为真阳性;HA基因阴性,其它三种基因阳性,为可疑阳性.结果 共检测3673份样本,其中,阴性2404份,甲型流感阳性样本共1269份,阳性率34.5％;阳性样本中季节性甲型流感641份,占50.5％;甲型H1N1流感阳性628份,占49.5％（真阳性433份,占68.9％,可疑阳性195份,占31.1％）.在整个流行季,真阳性与可疑阳性在相同时间的检出率比值基本不变.甲型H1N1检出高峰在10月,季节性甲流则分别在9月和12月.中青年人群的阳性检出率为57.5％,儿童为39.9％,而60岁以上的老年人仅占2.6％.结论 甲型H1N1流感以M、HA、NP、RNaesP四种基因阳性毒株为主,其流行高峰出现在10月份,感染人群以中青年和儿童为主.     

神经肽Y、降钙素基因相关肽、肿瘤坏死因子在先天性心脏病肺动脉高压中作用的研究

目的 本文着重阐述两种心血管调节肽-降钙素基因相关肽（CGRP）、神经肽Y（NPY）和先天性心脏病肺动脉高压（简称先心病肺高压）的关系;同时对先心病肺高压患儿测定肿瘤坏死因子（TNF）水平,以判断其与肺高压严重程度的关系;对部分病例外周血NPY、CGRP水平行术前,术后对照,以观察分流被纠正后其变化情况;另外,对外周血CGRP、NPY浓度作相关分析。以判断它们是否具有相关性。方法 运用放射免疫分析方法测定80例先天性心脏病患儿血清NPY、CGRP及TNF浓度,其中13例为非肺高压组,67例为肺高压组,将两组进行比较,进行方差分析,t检验。结果 非肺高压组与轻、中、重度肺高压组血浆NPY及CCRP浓度均存在显著性差异;非肺高压组与中、重度肺高压组血浆TNY存在显著性差异。与轻度肺高压组无显著性差异;手术前后轻,中度肺高压组NPY和CGRP浓度存在显著性差异。而重度肺高压组差异不明显;NPY与CGRP存在负相关关系。结论 三种活性物质在先天性心脏病肺动脉高压的发生机制中有重要作用。NPY/CGRP比值的升高参与和促进了肺动脉高压的形成和发展。     

Identification of Two Homozygous Sequence Variants in the COL7A1 Gene Underlying Dystrophic Epidermolysis Bullosa by Whole‐Exome Analysis in a Consanguineous Family

Dystrophic epidermolysis bullosa (DEB) is an inherited skin disorder with variable severity and heterogeneous genetic involvement. Diagnostic approaches for this condition include clinical evaluations and electron microscopy of patients’ skin biopsies, followed by Sanger sequencing (SS) of a large gene (118 exons) that encodes the alpha chain of type VII collagen (COL7A1) located on Chromosome 3p21.1. However, the use of SS may hinder diagnostic efficiency and lead to delays because it is costly and time‐consuming. We evaluated a 5‐generation consanguineous family with 3 affected individuals presenting the severe generalised DEB phenotype. Human whole‐exome sequencing (WES) revealed 2 homozygous sequence variants: the previously reported variant p.Arg578* in exon 13 and a novel variant p.Arg2063Gln in exon 74 of the COL7A1 gene. Validation by SS, performed on all family members, confirmed the cosegregation of the 2 variants with the disease phenotype. To the best of our knowledge, 2 homozygous COL7A1 variants have never been simultaneously reported in DEB patients; however, the upstream protein truncation variant is more likely to be disease‐causing than the novel missense variant. WES can be used as an efficient molecular diagnostic tool for evaluating autosomal recessive forms of DEB.