先天性视网膜劈裂症一家系基因突变位点分析

目的 对一个先天性视网膜劈裂(XLRS)家系进行基因分析,观察其视网膜劈裂基因（RS1基因）的突变位点。方法 家系研究。一个三代15人的XLRS家系纳入研究。包括先证者在内的9名家系成员行眼科常规检查。6名家系成员同时行眼底彩色照相、光相干断层扫描检查。其中,男性6人10只眼,女性3人6只眼。抽取家系成员中12人的外周静脉血,采用聚合酶链反应方法,对RS1基因的6个外显子各片段进行扩增后直接DNA测序分析,明确基因突变位点。结果 眼底检查者正常4人8只眼,其中男性1人2只眼,女性3人6只眼。均无视力下降主述。检查结果符合XLRS临床诊断者5例10只,均为男性。患眼视力手动～0.5。出现视力下降时年龄均＜10岁。眼底彩色照相检查可见黄斑区放射状囊样劈裂、周边视网膜劈裂;OCT检查可见黄斑区劈裂8只眼,周边视网膜劈裂6只眼。基因检测结果显示,正常者3人,男性。5例临床确诊者RS1基因在外显子4的末位碱基发生错义突变（c.326G＞T）,该突变导致RS1基因编码的视网膜劈裂蛋白第109位氨基酸由甘氨酸变为缬氨酸（p.Gly109Val）。1名3岁儿童具有与5例患者相同的基因突变。眼底检查正常的3名女性为p.Gly109Val突变的杂合子,为致病基因携带者。结论 p.Gly109Val突变为XLRS家系的一个新的突变位点。     

中国人X连锁遗传性视网膜劈裂症患者RS1基因突变分析及临床特征

目的通过对国人X连锁遗传性视网膜劈裂症(XLRS)患者RS1基因突变分析,描述RS1基因突变特点和XLRS表型特征。设计回顾性病例系列。研究对象本实验室收集XLRS患者27例,其中12例家族史明确,15例为散发。方法利用PCR扩增DNA直接测序方法检测27例患者RS1基因6个编码外显子。并对所有患者行详细的眼科检查,包括矫正视力、裂隙灯显微镜查眼前节、散瞳后直接眼底镜查眼底及眼底照相、相干光断层扫描(OCT)和视网膜电图(ERG)。主要指标RS1基因基因突变,发病年龄,视力,眼底表现。结果在27例患者中检测到27种RS1基因突变,其中4种为新发现突变,22种位于第4-6外显子(85.2%),即RS1蛋白的盘状结构域。27种突变包括15种错义突变(55.6%),4种无义突变(14.8%),4种缺失或插入(14.8%),2种剪接位点改变(7.4%),2种大片段缺失(7.4%)。27例患者均为男性,平均发病年龄(4.70±1.25)岁(0~34岁)。平均最佳矫正视力(0.22±0.28)(手动~1.0)。27例患者(54眼)中,40眼黄斑劈裂(74.1%),7眼仅有周边视网膜劈裂,5眼黄斑萎缩(其中3眼合并周边视网膜劈裂)。结论本研究结果扩大了RS1基因突变谱,第4-6外显子区域是XLRS患者RS1基因突变的热突变区域,对怀疑为XRLS患者应先进行RS1基因第4-6外显子区域测序。     

玻璃体视网膜手术治疗先天性视网膜劈裂及其并发症的疗效观察

目的 观察玻璃体视网膜手术治疗先天性视网膜劈裂(XLRS)及其并发视网膜脱离和(或)玻璃体积血的疗效.方法 回顾分析接受玻璃体视网膜手术治疗的XLRS并发视网膜脱离和(或)玻璃体积血患者21例27只眼的临床资料.所有患眼眼底及光相干断层扫描(OCT)检查均发现黄斑微囊样劈裂病变合并周边部视网膜劈裂.平均视力0.11±0.09,黄斑劈裂平均面积为(1.09±0.56) mm2.12只眼并发孔源性视网膜脱离,5只眼并发牵拉性视网膜脱离,6只眼并发玻璃体积血,4只眼同时合并视网膜脱离和玻璃体积血.均行经扁平部三通道闭合式玻璃体切割手术.根据情况行内界膜剥离,眼内激光光凝,C3 F8或硅油填充.手术后随访9～122个月,平均随访时间51个月.观察视力以及视网膜解剖结构改善情况.结果 末次随访视力提高者20只眼,占74.1％;维持不变者7只眼,占25.9％.平均视力提高至0.26±0.15.与治疗前平均视力比较,差异具有统计学意义(t=-6.320,P=0.000).27只眼视网膜解剖结构复位良好,视网膜平伏.OCT检查显示,黄斑劈裂平均面积(0.29±0.21) mm2,较治疗前黄斑劈裂平均面积显著缩小(t=10.358,P=0.000);黄斑微囊样病变得到明显的改善.随访期间4只眼出现并发症,占14.8％.其中,2只眼分别在手术后6、8个月并发增生性玻璃体视网膜病变伴牵拉性视网膜脱离;1只眼在手术后4个月出现并发性白内障;1只眼在手术后15个月因新发视网膜裂孔而发生玻璃体积血.给予再次手术治疗后,4只眼视网膜复位良好.结论 玻璃体视网膜手术能有效提高XLRS患者视力,恢复视网膜解剖结构,获得良好的治疗效果.     

先天性视网膜劈裂症基因突变型与临床表现型的关系

背景研究表明,导致先天性视网膜劈裂症（XLRS）的原因是RS1基因突变,XLRS的不同临床表现型与不同的基因突变类型有关,但基因型与表现型的关系仍不十分清楚。目的研究12个中国人XLRS家系及检测到的11种XLRS!基因突变与临床表现型的关系。方法对12个XLRS家系的28例男性患者（其余3例已死亡）及女性携带者和正常对照者分别进行详细的眼科检查,包括视力、屈光度、裂隙灯及眼底检查,部分患者行全视野视网膜电图（ERG）、眼底血管造影、光学相干断层扫描（OCT）及A／B型超声检查。采用聚合酶链反应（PCR）和单链构象多态性（SSCP）分析,对XLRSl基因突变进行筛查,并对发现异常泳带的PCR产物进行DNA测序,以明确突变位点及突变类型。结果12个家系的28例男性患者中27例有典型的黄斑及视网膜劈裂表现,ERG检查可见b波振幅下降,b／a波比值倒置。12个家系中检出11种不同的XLRSl致病基因突变,其中有4种新发现的基因突变,即：位于外显子1的移码突变（L9CfsX20）;位于外显子5的Aspl45His,Argl56Gly和Trp163X突变,并新发现1种非疾病相关多态性（NSP）,即位于外显子6的576C—T（Pr0192Pro）改变。同时发现位于外显子1的移码突变（22de／T）,外显子1与内含子1剪接位点突变（IVS1＋2TtoC）和Argl02Gln,Arg209His,Arg213Gln突变家系的患者,临床表现为严重型XLRS。结论XLRS1基因突变是导致中国人先天性视网膜劈裂的原因,XLRS的严重临床表现与基因突变的类型及突变位点具有一定的相关性。     

先天性视网膜劈裂症的RS1基因分析

目的观察先天性视网膜劈裂症的临床特征,研究患者的基因突变。方法采用聚合酶链反应（PCR）法,对先天性视网膜劈裂症一家系6名成员RS1基因的6个外显子各片段进行扩增,直接进行DNA测序分析,以明确突变位点及突变类型。结果先证者发生第625位核苷酸C→T,导致第209位氨基酸Arg→Cys。家系中有2名女性成员的外显子6核苷酸第625位的单碱基突变,为无临床症状和体征的突变携带者。结论本家系中先天性视网膜劈裂症患者是由RS1基因突变所致。     

先天性视网膜劈裂症家系RS1基因的突变筛查分析

先天性视网膜劈裂症(XLRS)又称X连锁隐性遗传病,是引起男性青少年黄斑变性的最主要原因[1,2]。其临床表现为眼底病变处视网膜神经纤维层裂开,玻璃体积血及玻璃体内一半透明的膜[1]。1997年Sauer等[3]将导致XLRS发病的视网膜劈裂症基因（RS1基因）定位于Xp22.2。我们在临床工作中发现XLRS2家系,通过聚合酶链反应(PCR)基因扩增联合DNA测序,对其进行了基因检测,以明确突变位点及突变类型,现将结果报道如下。1对象和方法临床确诊的XLRS患者6例和无眼疾的50名正常对照人群纳入研究。本研究遵循赫尔辛基宣言,并经临沂市人民医院伦理委员会审核批准,所有受检者均已签署书面知情同意书。     

先天性视网膜劈裂症基因突变的分析研究

目的 研究中国先天性视网膜劈裂症（XLRS）患者的基因突变，为患者及亲属提供基因诊断及遗传咨询。 方法 采用聚合酶链反应（PCR）方法，对12个XLRS家系29位男性患者及女性携带者和100名正常对照者的XLRS1基因的6个外显子各片段进行扩增，应用单链构像多态性（SSCP）分析, 对PCR产物进行基因突变筛查，并对发现异常泳动带的PCR产物进行DNA测序, 以明确突变位点及突变类型。 结果 12个XLRS家系检出11种不同的XLRS1致病基因突变，其中，第一外显子碱基缺失致移码突变1种: c.22delT（L9CfsX20）；1种碱基缺失致无义突变（Trp163X）；1种剪接位点突变(c.52+2 T→C; IVS1+2T to C)； 8种碱基替换导致错义突变（Ser73Pro、Arg102Gln、Asp145His、Arg156Gly、Arg200Cys、Arg209His、Arg213Gln、Cys223Arg)。对照者未检测到基因突变。新发现4种基因突变，即：移码突变（L9CfsX20），位于外显子5的Asp145His、Arg156Gly、Trp163X突变；一种新发现的非疾病相关多态性（NSP)：即位于第6外显子的 c.576C→T (Pro192Pro)改变。 结论 12个家系发现11种不同的XLRS1致病基因突变，XLRS1基因突变是导致中国XLRS症的原因。基因突变检测可以为患者及亲属提供基因诊断及家系遗传指导。 (中华眼底病杂志, 2006, 22: 77-81)     

先天性视网膜劈裂症基因突变的分析

目的 研究先天性视网膜劈裂症（XLRS）家系的XLRS1基因突变情况及其发病机制，为建立基因诊断的方法提供理论依据。 方法 采用聚合酶链反应（PCR）和DNA直接测序方法对两个家系的XLRS1基因编码区进行突变的筛选及检测。 结果 在家系1中发现Pro193Ser突变。 结论 在XLRS家系中发现XLRS1基因突变。本研究结果可直接应用于XLRS 的遗传咨询和产前基因诊断。 （中华眼底病杂志，2004，20：149-151）     

高度近视眼黄斑视网膜劈裂光相干断层扫描观察

目的 观察高度近视眼黄斑视网膜劈裂的光相干断层扫描(OCT)图像及临床特征.方法 回顾性分析经最佳矫正视力、屈光度、前置镜、A/B型超声和OCT检查确诊的154例高度近视患者158只眼的临床资料.OCT检查主要进行黄斑部水平及垂直线性扫描.根据OCT检查后极部是否存在视网膜劈裂,将其分为劈裂组、非劈裂组.其中,劈裂组53例55只眼,占34.8%;非劈裂组101例103只眼,占65.2%.回顾分析时,对两组患者年龄、性别、屈光度、视力、眼轴长度、后巩膜葡萄肿发生率、玻璃体牵引率、视网膜脱离发生率进行比较.结果 B型超声检查显示,158只眼均有后巩膜葡萄肿.OCT检查显示,视网膜劈裂组55只眼中,内层劈裂15只眼,占27.3%;外层劈裂53只眼,占96.4%;中层劈裂7只眼,占12.7%.内外层劈裂可单独存在,也可共存于同一患眼,中层劈裂都与外层劈裂相伴.存在2种或以上类型的劈裂13只眼,占23.6%;单纯外层劈裂40只眼,占72.7%;仅有内层劈裂2只眼,占3.6%.伴视网膜脱离26只眼,占47.3%;伴黄斑裂孔13只眼,占23.6%;伴玻璃体牵引12只眼,占21.8%.非劈裂组103只眼中,伴玻璃体牵引23只眼,占22.3%;黄斑裂孔19只眼,占18.4%;视网膜脱离21只眼,占20.4%.两组患者年龄(t=0.250)、屈光度(t=1.156)、眼轴(t=0.252)、性别(X2=1.075)构成、黄斑裂孔(X2=0.598)、后巩膜葡萄肿(X2=∞)、玻璃体牵引(X2=0.05)的发生率比较.差异均无统计学意义(P>0.05);但视网膜劈裂组视力明显低于非劈裂组(X2=6.345,P<0.05);视网膜脱离的发生率明显高于非劈裂组(X2=12.400,P<0.05).结论高度近视眼黄斑视网膜劈裂存在于视网膜内中外多个层次,其中外层劈裂最常见;患者往往视力低下且常伴发其他黄斑病变,以视网膜脱离最常见.     

高度近视眼黄斑视网膜劈裂光相干断层扫描观察

目的 观察高度近视眼黄斑视网膜劈裂的光相干断层扫描(OCT)图像及临床特征.方法 回顾性分析经最佳矫正视力、屈光度、前置镜、A/B型超声和OCT检查确诊的154例高度近视患者158只眼的临床资料.OCT检查主要进行黄斑部水平及垂直线性扫描.根据OCT检查后极部是否存在视网膜劈裂,将其分为劈裂组、非劈裂组.其中,劈裂组53例55只眼,占34.8%;非劈裂组101例103只眼,占65.2%.回顾分析时,对两组患者年龄、性别、屈光度、视力、眼轴长度、后巩膜葡萄肿发生率、玻璃体牵引率、视网膜脱离发生率进行比较.结果 B型超声检查显示,158只眼均有后巩膜葡萄肿.OCT检查显示,视网膜劈裂组55只眼中,内层劈裂15只眼,占27.3%;外层劈裂53只眼,占96.4%;中层劈裂7只眼,占12.7%.内外层劈裂可单独存在,也可共存于同一患眼,中层劈裂都与外层劈裂相伴.存在2种或以上类型的劈裂13只眼,占23.6%;单纯外层劈裂40只眼,占72.7%;仅有内层劈裂2只眼,占3.6%.伴视网膜脱离26只眼,占47.3%;伴黄斑裂孔13只眼,占23.6%;伴玻璃体牵引12只眼,占21.8%.非劈裂组103只眼中,伴玻璃体牵引23只眼,占22.3%;黄斑裂孔19只眼,占18.4%;视网膜脱离21只眼,占20.4%.两组患者年龄(t=0.250)、屈光度(t=1.156)、眼轴(t=0.252)、性别(X2=1.075)构成、黄斑裂孔(X2=0.598)、后巩膜葡萄肿(X2=∞)、玻璃体牵引(X2=0.05)的发生率比较.差异均无统计学意义(P>0.05);但视网膜劈裂组视力明显低于非劈裂组(X2=6.345,P<0.05);视网膜脱离的发生率明显高于非劈裂组(X2=12.400,P<0.05).结论高度近视眼黄斑视网膜劈裂存在于视网膜内中外多个层次,其中外层劈裂最常见;患者往往视力低下且常伴发其他黄斑病变,以视网膜脱离最常见.     

Genotypic and phenotypic spectrum of X-linked retinoschisis in Australia

BACKGROUND: X-linked retinoschisis (XLRS), an X-linked recessive inherited degenerative retinopathy, is characterized by splitting in the nerve fibre layer and is caused by alterations in the RS1 gene. The aim of the present study was to review both the phenotypic features of XLRS and the mutation spectrum of the RS1 gene in an Australian cohort. METHODS: Patients were recruited from ophthalmic and paediatric hospitals as well as private ophthalmic clinics across Australia. A cohort of 18 presumably unrelated families was identified. Twenty-two affected patients underwent clinical examination. Following DNA extraction all six exons of the RS1 gene were sequenced. RESULTS: The median age at diagnosis was 8 years (range 1-43 years); the median age at review was 14 years (range 5-63 years). The median best-corrected visual acuity upon review was 6/24 (range 6/6-1/36). Typical foveal schisis was found in 90.1% eyes examined (39/43) while peripheral schisis was present in 30% of eyes (13/43). The scotopic blue b-wave amplitude ranged between 2% and 82% of the mean normal amplitude. Five novel mutations (61G-->T, Gly21X; 103C-->T, Gln35X; 327-329del, Cys110del; 527T-->C, Phe176Ser; 573Gdel, Pro192fs) and six previously identified missense mutations (304C-->T, Arg102Trp; 305G-->A, Arg102Gln; 336G-->C, Trp112Cys; 418G-->A, Gln140Arg; 598C-->T, Arg200Cys; 625C-->T, Arg209Cys) were found. The mutations present in codons 21 and 102 were each identified in two presumably unrelated pedigrees. One previously described point deletion (416Adel) was identified. Two pedigrees contained affected individuals where exons 2 or 3, respectively, were unable to be amplified, indicating the likely presence of a significant deletion. No mutation was found in the RS1 gene in two affected individuals from different pedigrees. CONCLUSION: Population genetic studies of XLRS have not previously been conducted in Australia. The phenotype associated with these mutations varied. The identification of each pedigree's specific mutation allows future determination of female carrier status for genetic counselling purposes. Further study into the refinement of the XLRS phenotype as well as the degree of intrafamilial phenotypic variation is required.     

Clinical features of X linked juvenile retinoschisis associated with new mutations in the XLRS1 gene in Italian families

AIMS: To describe the clinical phenotype of X linked juvenile retinoschisis in eight Italian families with six different mutations in the XLRS1 gene. METHODS: Complete ophthalmic examinations, electroretinography and A and B-scan standardised echography were performed in 18 affected males. The coding sequences of the XLRS1 gene were amplified by polymerase chain reaction and directly sequenced on an automated sequencer. RESULTS: Six different XLRS1 mutations were identified; two of these mutations Ile81Asn and the Trp122Cys, have not been previously described. The affected males showed an electronegative response to the standard white scotopic stimulus and a prolonged implicit time of the 30 Hz flicker. In the families with Trp112Cys and Trp122Cys mutations we observed a more severe retinoschisis (RS) clinical picture compared with the other genotypes. CONCLUSION: The severe RS phenotypes associated with Trp112Cys and to Trp122Cys mutations suggest that these mutations determine a notable alteration in the function of the retinoschisin protein.     

Novel <Emphasis Type="Italic">XLRS1</Emphasis> gene mutations cause X-linked juvenile retinoschisis in Chinese families

Purpose To investigate various XLRS1 (RS1) gene mutations in Chinese families with X-linked juvenile retinoschisis (XLRS or RS). Methods Genomic DNA was isolated from leukocytes of 29 male patients with X-linked juvenile retinoschisis, 38 female carriers, and 100 normal controls. All 6 exons of the RS1 gene were amplified by polymerase chain reaction, and the RS1 gene mutations were determined by direct sequencing. Results Eleven different RS1 mutations in 12 families were identified in the 29 male patients. The mutations comprised eight missense, two frameshift, and one splice donor site mutation. Four of these mutations, one frameshift mutation (26 del T) in exon 1, one frameshift mutation (488 del G) in exon 5, Asp145His and Arg156Gly in exon 5, have not been previously described. One novel non-disease-related polymorphism, 576C to T (Pro192Pro) in exon 6, was also found. Six recurrent mutations, Ser73Pro and Arg102Gln mutations in exon 4 and Arg200Cys, Arg209His, Arg213Gln, and Cys223Arg mutations in exon 6, were also identified in this study. Conclusion RS1 gene mutations caused X-linked juvenile retinoschisis in these Chinese families.     

Surgical outcome after phototherapeutic keratectomy in patients with TGFBI-linked corneal dystrophies in relation to molecular genetic findings

Purpose  To evaluate the correlation between surgical outcome after phototherapeutic keratectomy in patients with autosomal dominant transforming growth factor, beta-induced (TGFBI)-linked corneal dystrophies (CD) and molecular genetic findings regarding the TGFBI gene. Methods  Twelve patients were examined to investigate genotype by direct sequencing of the TGFBI gene. Twenty eyes of 12 patients were treated with phototherapeutic keratektomy (PTK) to remove superficial corneal opacifications and to decrease recurrent erosions. Surgical outcome, including visual improvement, recurrence of opacifications, postoperative complications, and additional therapeutic proceedings were reported and compared with the molecular genetic results. Results  Four different missense mutations were identified within the coding region of the TGFBI gene: Arg124Cys in one eye, Arg555Trp in nine eyes, Arg124His in four eyes and Gly623Arg in six eyes. In all eyes the PTK was successful without clinically significant recurrent opacifications after a mean follow-up time of 17.6 months (min 3 months, max 42 months). The best corrected visual acuity (BCVA) improved with an average increase of 3.1 lines (minimum 2 lines, maximum 5 lines). In one eye (Arg124Cys), we observed delayed wound healing and a delayed increase in BCVA, in two eyes we performed an Epilasik to correct remaining hyperopia, and in four eyes we fitted rigid gas-permeable tricurve contact lenses to correct the remaining irregular astigmatism. Conclusions  The variable genotypes in patients with TGFBI-linked corneal dystrophies lead to significantly different results after surgical treatment. The Gly623Arg mutation seems to be an optimum genotype on which to perform PTK even in older patients. It is essential to determine the genotype in order to standardize the PTK treatment and to evaluate the success in TGFBI-linked corneal dystrophies.     

Three novel mutations in the X-linked juvenile retinoschisis (XLRS1) gene in 6 Japanese patients, 1 of whom had Turner's syndrome

We examined the XLRS1 gene for mutations in 6 Japanese patients with X-linked juvenile retinoschisis from a total of three families (5 males and 1 female), and from 3 obligate carrier females. DNA was amplified for all six coding exons of the XLRS1 gene with established primer pairs, and was sequenced directly. Each family had a different mutation, Trp96stop, 522+1g-->a, and Lys167Asn in the XLRS1 gene. Affected patients had a hemizygous mutant allele while the obligate carrier females were heterozygotes who had both wild-type and mutant-type alleles. A proband female, who was the offspring of asymptomatic and nonconsanguineous parents, was found to have a chromosomal karyotype (45, X) that was indicative of Turner's syndrome. These three different mutations in the XLRS1 gene have not been previously reported. Further studies are needed to determine the relationship between these defects in the XLRS1 gene and the phenotypic expression of the disease.     

Two cases of X-linked juvenile retinoschisis with different optical coherence tomography findings and RS1 gene mutations

The optical coherence tomography (OCT) findings, clinical features, and mutations in the RS1 gene of two unrelated patients with X-linked retinoschisis (XLRS) are reported herein. Two Chinese patients with early onset XLRS were given a comprehensive ophthalmologic examination and OCT investigation. The RS1 gene was screened for sequence alterations in all exons and splice regions. The two patients presented with different phenotypic features and OCT findings. One patient with more severe clinical presentation had a RS1 exon 1 deletion and a P193S mutation was found in the other patient with mild macular involvement. OCT demonstrates the markedly different features of XLRS patients with different RS1 mutations. This study strengthens the role of OCT in the diagnosis and monitoring of XLRS.