目的 探讨常见过敏原致过敏反应易感性与过敏性皮肤病患者外周血HLA DRB1*0901(rs2395185)基因多态性的关系.方法 选择广州地区皮肤科就诊患者304例,采用免疫印迹法检测患者血清19种过敏原,PCR测序检测外周血HLA DRB1*0901位点基因型,分析HLA DRB1*0901基因多态性与过敏原致过敏反应发病风险的关系.结果 户尘螨过敏原检测结果阳性、阴性患者间rs2395185的G/G基因型、G/T基因型分布差异有统计学意义(P<0.05),G/G、G/T基因型与户尘螨过敏的发病风险有关(OR=2.940,95％CI:1.167～7.407;OR=2.889,95％CI:1.136～7.348).轻度过敏与中重度过敏患者的G/T基因型分布差异有统计学意义(P<0.05),G/T基因型是过敏原反应严重程度的危险因素(OR=8.681,95％CI:1.077～69.989).结论 广州地区居民户尘螨皮肤过敏发病风险可能与其外周血HLA DRB1*09:01(rs2395185)位点G/G、G/T多态性有关,19种过敏原过敏的严重程度可能与rs2395185位点G/T多态性有关.

BackgroundThe presence of more than one polyQ-related gene within a single individual is a rare incidence, which may provide the potential opportunity to study the combined effects of these spinocerebellar ataxia (SCA) genes. MethodsWe retrospectively analyzed genetic data from 112 SCA3 probands and found Patient 1 harbored expanded ATXN2 allele (33 repeats) and intermediate TBP allele (41 repeats), and Patient 2 with intermediate ATXN2 allele (32 repeats). Detailed clinical and oculomotor performances were investigated. The age at onset and oculomotor parameters of both patients were compared with matched pure SCA3 groups controlling either disease severity or CAG repeats. ResultsMost of the clinical phenotypes and oculomotor characteristics of these two patients were common to typical SCA3 patients. Compared to pure SCA3 groups controlling disease severity, mild reduced horizontal saccade velocity could be detected in both patients. However, mild expansions of the ATXN2 allele seemed to have no influence on the age at onset of Patient 1 but might have a mild impact on Patient 2. ConclusionOur study provides supporting evidence that mild expansions of ATXN2 may have modifying effects on SCA3 phenotype. Larger control series and longitudinal data are warranted to confirm our results.

目的 分析一马凡综合征(Marfan syndrome,MFS)家系的临床特征,进行原纤维蛋白-1基因(FBN1)突变分析,明确该家系的致病基因,以加强对此病的认识,提高早期诊断率,并对1例MFS孕妇进行产前诊断.方法 描述一家系8例患者的临床表现,提取先证者及其家族成员外周血DNA,用PCR和DNA测序技术检测FBN1基因外显子中的潜在突变.提取MFS孕妇羊水细胞DNA和培养后羊水细胞的RNA,RT-PCR扩增RNA检测该家系所发现的阳性位点.结果 该家系的遗传方式为常染色体显性遗传,4代共有8例患者,其中4例在47～59岁猝死,先证者和其他3例存活患者均具有心血管系统的异常(主动脉根部夹层或主动脉根部扩张)、晶状体脱位、高度近视及MFS的特征性骨骼改变,并随着年龄的增长而逐步加重.基因检测显示先证者FBN1基因54号外显子存在c.6554T>C(p.Ile2185Thr)错义突变,其他3例存活患者均发现该位点突变,而表型正常的成员未发现该突变.胎儿羊水细胞的DNA与羊水培养细胞RNA均发现该位点的突变.结论 FBN1基因错义突变c.6554T>C(p.Ile2185Thr)为该家系致病的分子基础,基因检测有助于早期明确诊断以便尽早干预治疗,避免夹层动脉破裂导致猝死的不良结局.该MFS孕妇的胎儿遗传有该FBN1的致病突变,给双亲决定是否终止妊娠提供参考,有助于患病家族的优生优育.

OBJECTIVE:To detect potential variation in glutaryl-CoA dehydrogenase (GCDH) gene among three Chinese families affected with glutaric acidemia type Ⅰ(GA-1) and correlate the genotypes with phenotypes.METHODS:Genomic DNA was extracted from peripheral blood samples derived from three patients with GA-1 and their family members. The coding regions of the GCDH gene were amplified with PCR and subjected to Sanger sequencing.RESULTS:The clinical manifestation of the patients varied from macrocephaly to severe encephalopathy, with notable phenotypic difference between siblings carrying the same variation. In pedigrees 1 and 2, the probands have carried compound heterozygous variations c.1133C>T(p.Ala378Val) and c.1244-2A>C, which were derived their fathers and mothers, respectively. In pedigree 3, the proband has carried compound heterozygous variation c.339delT (p.Tyr113) and c.406G>T (p.Gly136Cys). Among these, variations c.339delT and c.1133C>T were verified as novel by retrieval of dsSNP, HGMD and 1000 genome database. Bioinformatic analysis suggested that above variations can affect protein function and are probably pathogenic.CONCLUSION:Above discovery has expanded the mutation spectrum of the GCDH gene. No correlation was found between the clinical phenotype and genotype of GA-1 patients.

OBJECTIVE:To delineate the clinical features and potential mutation of the ATP7A gene in a family affected with Menkes disease.METHODS:Clinical data of a patient and his family members were analyzed. Sanger sequencing and multiplex ligation-dependent probe amplification (MLPA) assays were performed to detect the mutation of the ATP7A gene.RESULTS:The patient was admitted at the age of 5 months due to severe epilepsy and marked delayed psychomotor development. Significantly light complexion, pudgy cheeks and sparse fuzzy wooly hair were noted. Cranial magnetic resonance imaging and angiography revealed cortical atrophy, leukoencephalopathy and circuitous of intracranial vessels. The plasma ceruloplasmin was decreased. MLPA has identified a deletion spanning exons 8 to 12 of the ATP7A gene. His mother was found to be a heterozygous carrier of the same mutation.CONCLUSION:The clinical features and a novel mutation of the ATP7A gene of the family have been delineated.

Objective To investigate the NOTCH3 gene locus, clinical manifestation and MRI features of autosomal dominant hereditary cerebral arteriopathy with subcortical infarcts and white matter encephalopathy (CADASIL). Methods Genetic testing were performed in 831 cases of suspected CADASIL from 2009 to 2016, and 21 patients with CADASIL were selected as CADASIL group from NOTCH3 positive patients from Xinhui Hospital or Nanfang Hospital affiliated to Southern Medical University. Another 32 patients with CADASIL score of more than 14 points served as CADASIL-like group. The clinical features and magnetic resonance imaging results of the two groups were analyzed. Results We have screened out 275 patients carrying NOTCH3 gene in 831 cases, with total of 71 mutant sites, in which 100 patients carried excon 4 mutant and 97 patients carried excon 11 mutant. Excon 11 p.R544C in Notch3 gene was found to be the maximum frequency of mutant site with mutation rate of 23.27%. White matter high signal intensity (WMHs) was the common expression in iconography and almost showed in all patients (95.2% in CADASIL group and 100% in CADASIL-like group). WMHs was always bilateral and symmetric. The rates of external capsule involvement in two groups were high with no statistically significant difference (71.4% in CADASIL group vs 71.9% in CADASIL-like group,P >0.05). However, temporal pole involvement rate in CADASIL group was statistically significantly higher than that in CADASIL-like group (42.9% vs 12.5%, P < 0.05).The occurrence rates of multi-lacunar infarcts were 95.2% in CADASIL group and 90.6%in CADASIL-like group (P < 0.05). Cerebral microbleeds were observed in 76.2% (16/21) cases in CADASIL group and 46.9%(15/32) cases in CADASIL-like gruop with significant difference (P < 0.05). Conclusion In our study, NOTCH3 mutations most commonly occur in exon 4 and exon 11,and R544C in exon 11 accounted for nearly one fourth of the pedigrees, In CADASIL cases, bilateral, and symmetrical white matter lesions widely exist, with frequent involvement of temporal pole and cerebral microbleeds.

目的：探讨2例丙酸血症患儿的临床特征及基因突变特点。方法回顾性分析2例丙酸血症患儿的病例资料及家系的基因突变检测，并复习相关文献，分析本病的临床表现、生化检测和基因突变特点。结果例1患儿，男，11 d，因“呕吐、气促”3 d 入院，新生儿期发病，入院后遗传代谢病筛查发现丙酰肉碱增高，丙酰肉碱／乙酰肉碱增高，尿有机酸分析发现甲基枸橼酸、甲基巴豆酰甘氨酸、3-羟基丙酸增高，诊断为丙酸血症，治疗过程中表现为反复感染，骨髓抑制，生物素治疗有效，随访过程中发现丙酰基肉碱水平较高，高甘氨酸血症，限制异亮氨酸、缬氨酸、蛋氨酸和苏氨酸饮食后丙酰基肉碱下降的同时易发生支链氨基酸的缺乏；虽然积极治疗仍表现明显的精神运动发育落后；基因检测患儿PCCB c．1301C＞T p．（Ala434Val）纯合突变，父母为PCCB c．1301C＞T p.（Ala434Val）杂合突变。例2患儿，女，7个月，因“呕吐伴代谢性酸中毒”入院。婴儿期发病，入院经检测诊断丙酸血症。患儿表现较例1患儿轻微，无反复感染，无骨髓抑制。随访过程中发现丙酰基肉碱水平较例1患儿明显偏低，有高甘氨酸血症，限制异亮氨酸、缬氨酸、蛋氨酸和苏氨酸饮食后丙酰基肉碱下降的同时不易发生支链氨基酸的缺乏；运动发育正常，精神及语言发育稍落后，检测到 PCCB 基因的一个纯合突变：c．167＿179del13insC p.（Asp56＿Lys60delinsAla），其父亲、母亲各检测到一个杂合突变。结论2例丙酸血症患儿中，新生儿期发病、反复感染、骨髓抑制、高丙酰基肉碱水平，饮食控制中易发生支链氨基酸的缺乏，提示病情预后不佳，容易发生精神运动发育落后；目前报道2例（含本例）基因突变为 PCCB c.1301C＞T p．（Ala434Val）纯合突变，均病情严重。

目的 探讨2例丙酸血症患儿的临床特征及基因突变特点。 方法 回顾性分析2例丙酸血症患儿的病例资料及家系的基因突变检测，并复习相关文献，分析本病的临床表现、生化检测和基因突变特点。 结果 例1患儿，男，11 d，因呕吐、气促 3 d入院，新生儿期发病，入院后遗传代谢病筛查发现丙酰肉碱增高，丙酰肉碱/乙酰肉碱增高，尿有机酸分析发现甲基枸橼酸、甲基巴豆酰甘氨酸、3-羟基丙酸增高，诊断为丙酸血症，治疗过程中表现为反复感染，骨髓抑制，生物素治疗有效，随访过程中发现丙酰基肉碱水平较高，高甘氨酸血症，限制异亮氨酸、缬氨酸、蛋氨酸和苏氨酸饮食后丙酰基肉碱下降的同时易发生支链氨基酸的缺乏；虽然积极治疗仍表现明显的精神运动发育落后；基因检测患儿PCCB c．1301C>T p．(Ala434Val)纯合突变，父母为PCCB c．1301C>T p．(Ala434Val)杂合突变。例2患儿，女，7个月，因呕吐伴代谢性酸中毒入院。婴儿期发病，入院经检测诊断丙酸血症。患儿表现较例1患儿轻微，无反复感染，无骨髓抑制。随访过程中发现丙酰基肉碱水平较例1患儿明显偏低，有高甘氨酸血症，限制异亮氨酸、缬氨酸、蛋氨酸和苏氨酸饮食后丙酰基肉碱下降的同时不易发生支链氨基酸的缺乏；运动发育正常，精神及语言发育稍落后，检测到PCCB基因的一个纯合突变：c.167_179del13insC p．(Asp56_Lys60delinsAla)，其父亲、母亲各检测到一个杂合突变。 结论 2例丙酸血症患儿中，新生儿期发病、反复感染、骨髓抑制、高丙酰基肉碱水平，饮食控制中易发生支链氨基酸的缺乏，提示病情预后不佳，容易发生精神运动发育落后；目前报道2例(含本例)基因突变为PCCB c．1301C>T p．(Ala434Val)纯合突变，均病情严重。

Objective To study the clinical, pathological and genetical manifestation of Gitelman syndrome.MethodThe clinical data, renal biopsy pathology, genetic mutations of patients diagnosed with Gitelman syndrome were analyzed.ResultsAll the patients showed hypopotassemia, hypomagnesemia, alkalipoisoning, hyperaldosteronemia,hyperreninemia.SLC12A3 complicated heterozygotic mutation was observed.Conclusions Gitelman syndrome in children is insidious and the incidence rate ofSLC12A3 complicated heterozygotic mutation is high. Genetic tests can diagnose the disease. Pediatrician must recognize the manifestations to avoid misdiagnosis.

Objective Gitelman Syndrome is a disease caused by the mutation of Na-Cl cotransporter gene(SLC12A3).The article studied the significance of diagnosis and identification by genetic mutation. Methods We collected the clinical data, then we sequenced the SLC12A3 gene by the first sequencing technology and MLPA. Results SLC12A3 complicated heterozygotic mutation was observed.One of them showed c.1964G>A, p.(Arg655His) and exon 8 deletion mutation, the other showed c.2543A>T, p.(Asp848Val) and c.976delG, p.(Val326fs) mutation of SLC12A3 gene in children. Conclusion The final diagnosis depended on gene diagnosis. Pediatrician must recognize the manifestations to advoid misdiagnosis.

Objective In order to make a well understanding on glycogen storage disease typeⅡ (GSDⅡ), this paper explored clinical features and genetic analysis of 7 patients with late-onset glycogen storage disease typeⅡ. Methods Clinical data of 7 patients with late-onset glycogen storage disease type Ⅱ were collected and acid α-glucosidase (GAA) gene sequencing was performed. Results Seven patients who belong to 4 families were at the age of 13-31 years old. The first symptom occurred at 6-17 years old, and the age at first and definitive diagnosis was 12-29 and 12-30 years old, respectively. The initial symptoms were mostly related to limb girdle muscular atrophy and weakness. The GAA activity ranged from 0 to 5.27 nmol/(mg·h). Sequencing analysis revealed 14 sequence variants, including 2 novel mutations (Q81X and c.1355_1356delC), 2 pseudodeficiency alleles (G576S and E689K), 8 polymorphic loci, and 2 sequence variants previously related with glycogen storage disease type Ⅱ pathogenesis (W746C and D645E). Conclusions Due to the apparently diagnostic delay, prognosis of patients with glycogen storage disease type Ⅱ could be improved by increasing the clinician's awareness of the disease. It is essential to combine clinical history with GAA activity and GAA gene analysis when we make a definitive diagnosis of glycogen storage disease type Ⅱ. Though siblings share the same set of GAA mutations, the phenotype regarding the course and severity of disease could vary substantially. doi: 10.3969/j.issn.1672-6731.2014.05.008

OBJECTIVE To review the clinical features of a families affected with glutaric acidemia type I (GA-1) and screen potential mutations in glutaryl-CoA dehydrogenase (GCDH) gene. METHODS Clinical data of the patients and their family members was analyzed. Genomic DNA was extracted from peripheral blood samples. The 11 exons and flanking sequences of the GCDH gene were amplified with PCR and subjected to direct DNA sequencing. RESULTS Two patients have manifested macrocephaly. Imaging analysis revealed arachnoid cyst and subdural effusion. The elder sister had encephalopathy crisis. The younger sister had significantly raised glutaric acid, whilst the elder sister was normal during the non-acute phase. Genetic analysis has revealed a homozygous c.1244-2A> C mutation of the GCDH gene in both patients. CONCLUSION The clinical features and mutation of the GCDH gene have been delineated in a Chinese family affected with GA-1. The c.1244-2A> C mutation may be particularly common in the Chinese population.

OBJECTIVE To evaluate the genetic diagnostic feasibility of Bruton's tyrosine kinase (BTK) gene in three families with X-linked agammagobulinemia (XLA) birth history, mutation analysis and prenatal genetic diagnosis of BTK gene for two families with XLA. METHODS Polymerase chain reaction (PCR) was applied to amplify the regions of exon and exon-intron boundaries of BTK gene in 3 unrelated patients of XLA and their mothers from January 2011 to June 2012. The PCR products were further analyzed by direct sequencing. Prenatal genetic diagnosis was performed by chorionic villus sampling after genotyping of mothers of probands. RESULTS Three novel mutations of BTK gene were identified in 3 pedigrees of XLA. A missense mutation c.1117C > A (p.L373I) were detected in pedigree 1. The mutation was possible damage by predicting in sillico. A nonsense mutation c.126T > G (p.Y42X) was found in pedigree 2. A single base deletion mutation c.1679delC (p. P560fsX10) was found in pedigree 3. The three mutations, p.L373I, p.Y42X and p. P560fsX10 were novel. The three novel mutations were absent in the 100 normal controls. The male fetus in pedigree 3 was free of mutations identical to the proband and the female fetus in pedigree 2 was a carrier. The two families continued the pregnancies and the infants showed no symptom of XLA after one year old. CONCLUSIONS Three novel mutations were identified. The mutations of p.Y42X and p. P560fsX10 in BTK gene may be the major causes of pedigrees 2 and 3 with XLA. The mutation p.L373I of BTK gene is possibly the cause of pedigree 1 with XLA, but functional verification is needed. For pedigree of XLA, direct sequencing of BTK gene is available for providing genetic counseling, prenatal diagnosis.

Objective To study the muscular pathological characteristics in riboflavin-responsive lipid storage myopathy before and after treatment. Methods A 10-year follow-up visit was made on a patient with riboflavin-responsive lipid storage myopathy, and the changes of serum enzymes, and both histological and ultrastructural data acquired by general muscular pathology, immunohistochemistry and electron microscope were observed before and after treatment by using levocarnitine. ETFDH gene were detected in the patient and his family. Results The patient presented limb weakness, difficulty in raising head and dysphagia, which were typical clinical features of lipid storage myopathy (LSM). The serum creatine kinase (CK) level and lactic dehydrogenase (LDH) level elevated evidently. EMG showed myogenic abnormality, and muscular pathology revealed numerous lipid droplets deposited in the fibers. ATPase staining showed predominant atrophy of typeⅠ fibers and relative increasing of the portion of typeⅡ fibers. Modified Gomori trichrome (MGT) staining did not observe ragged red fibers. Immunohistochemical staining showed positive expression of dystrophin. Sultan Ⅲ staining revealed multiple vacuolated myofibers. ETFDH gene test showed two heterozygous mutations in the patient. After treating with levocarnitine for several years, the patient could live a normal life. The muscular pathological result returned to normal. Conclusions After appropriate therapy, patient with lipid storage myopathy can not only gain complete remission clinically, but also the reversion of lesion pathologically. doi: 10.3969/j.issn.1672-6731.2014.06.005

Abstract Background: The aim of this study was to establish a sensitive method that can detect the presence of not only the common but also the unusual or unknown α-globin gene deletions for screening of α-thalassemia. We used quantitative multiplex PCR of short fluorescent fragments (QMPSF) for the α-globin genes (HBA) to screen α-thalassemia deletions. Methods: We set up and validated HBA-QMPSF using 50 negative and 100 positive controls of deletional α-thalassemia. To evaluate its ability to detect the presence of the common and unusual or unknown α-globin gene deletions, 579 unrelated samples were simultaneously analyzed using this assay and multiplex Gap polymerase chain reaction (Gap-PCR). The inconsistent results were further confirmed by multiplex ligation-dependent probe amplification (MLPA). Results: HBA-QMPSF was capable of detecting α-globin gene deletions with an acceptable variability as shown by mean values (SD) of allele dosage for the heterozygous deleted control obtained from intra- and inter-experimental replicates [0.63 (0.01) and 0.61 (0.03)]. In 572 out of the 579 unrelated subjects, HBA-QMPSF and multiplex Gap-PCR gave consistent results. In seven cases which were finally proved to be composed of one rare deletion --Thai/-α3.7, one novel deletion --SEA/-α2.8, four αααanti3.7/αα and one αααanti4.2/αα triplications, HBA-QMPSF showed deletion or duplication in the α-globin gene while multiplex Gap-PCR failed to give the correct diagnosis. Conclusions: HBA-QMPSF is able to detect the presence of the common and unusual or unknown α-thalassemia deletions and duplications. It can be used as an initial screening test for α-thalassemia caused by HBA gene copy number alteration.

Objective To report a DMD gene new mutation (c.9760_9781dup22/p.Pro3261LeufsX5) in a DMD children,and analyze his clinical manifestations.Methods Combining of MLPA and gene sequencing methods,we analyzed the DMD gene.Results The MLPA result showed that DMD gene exon 67 existed a diminutive mutation,and we used PCR and gene sequencing to confirm it,and find it to be a new mutation(c.9760_9781dup22/p.Pro3261LeufsX5)in DMD gene exon 67. Conclusions The mutation(c.9760_9781dup22/p.Pro3261LeufsX5) in DMD gene exon 67 is a new mutation,and this mutation is relevant to the DMD symptom and mental retardation.

Objective To improve the accuracy of detection of a single exon deletion and provide an accurate basis of genetic counseling and prenatal gene diagnosis of the family members. Methods 185 blood samples were collected and the DMD gene was detected between 2009 to 2010. DNA samples from the College of American Pathologists were used as negative control and positive control. A single exon deletion mutation in DMD gene was detected by MLPA, PCR and gene sequencing technologies.. Results In the term of 185 blood samples, the MLPA results showed that there existed single exon deletion mutations in DMD gene of 7 cases. PCR and gene sequencing were used to confirm, and a new mutation (c.9760_9781dup22/p.Pro3261LeufsX5) in DMD gene exon 67 was found just in one blood sample. Conclusion MLPA、PCR and gene sequencing technology are combined to improve the accuracy of detection of a single exon deletion and provide an accurate basis of genetic counseling and diagnosis of prenatal gene in the family members.

Objective To detecte a rare variant of [β43(CD2) Glu→Lys] at position 310 of theβ-globin gene previously unreported in the Hb variant carriers from a Chinese.Methods In phenotype analysis,standard hematological techniques were used to measure RBC counts and Hb concentration.PCR/ASO probe hybridization analysis,was used to scan 17 known types of beta-thalassemia mutation in Chinese.Using gene clone and DNA sequence analysis of the entire human β-globin gene to identify the underlying causative mutation of the sample and to identify its genotype.Results The proband presented a normal value with mean corpuscular volume(MCV,97fl) and elevated level of Hb(A_2+E)(46.99%),but known β-globin gene mutations were not found in the sample by reverse dot blot(RDB) analysis.DNA sequencing of theβ-globin gene region of the sample revealed heterozygosity for the G→A substitution at position 130 within exon 2 of theβ-globin gene,which was not reported in Chinese population previously.Conclusion [β43(CD2)Glu→Lys] is a rare variant of β-globin gene in Chinese.