Perinatal management and genetic testing and for a puerperant affected with ATP7B gene variation in Wilson disease / 中国综合临床

Objective:To explore the perinatal management of patients with WD during pregnancy, and to determine its genetic etiology and the possibility of fetal morbidity using the genetic detection of amniotic fluid and umbilical cord blood.Method:In terms of fine management during the perinatal period, a case of K-F ring was found in the Ophthalmology Department of Beijing Friendship Hospital, Capital Medical University in March 2019 due to eye astringency and eye swelling, and the hepatology department further diagnosed WD for one artificial abortion. After the second pregnancy in October 2020, multidisciplinary consultation and standardized treatment during pregnancy including gynecology and obstetrics, liver disease center, anesthesiology department, gastroenterology department and nutrition department were carried out. The genomes of patients' venous blood, amniotic fluid and umbilical cord blood were extracted and analyzed for ATP7B gene variation by Sanger sequencing.Result:Through multi-disciplinary collaborative management, the patient gave birth successfully in the case of pregnancy complicated with liver cirrhosis, portal hypertension, splenomegaly with hyperfunction, thrombocytopenia, anemia, esophageal and gastric varices and other complications. The phenotype of the newborn was normal, and the Apgar score was 10-10-10. Sequencing results showed that the patient had ATP7B p.Arg778Leu and p.Val890Met, which were missense heterozygous variants reported in the mutation database, and ACMG was classified as pathogenic variants. The results of amniotic fluid and umbilical cord blood showed that the fetus had only p.Arg778Leu single heterozygous variation, and it was predicted that there would be no clinical phenotype of WD.Conclusion:Perinatal multidisciplinary collaborative management has important protective significance for the successful pregnancy of patients with WD. Genetic screening of amniotic fluid and umbilical cord blood is conducive to early detection of fetal WD.

Enfermedad de Wilson: experiencia de un centro de referencia en Colombia / Wilson's disease: Experience of a reference center in Colombia

Resumen Introducción: la enfermedad de Wilson es una enfermedad heterogénea causada por mutaciones en el gen ATP7B. La presentación clínica es variable, en fenotipos hepáticos y neuropsiquiátricos. El objetivo de este estudio es describir una cohorte retrospectiva de pacientes. Materiales y métodos: estudio retrospectivo descriptivo de pacientes atendidos en el Hospital Pablo Tobón Uribe desde enero de 2004 a septiembre de 2017. Resultados: se reportaron 27 pacientes, 17 hombres y 10 mujeres. El tiempo de seguimiento medio fue de 2,18 años, el 40% presentó síntomas neurológicos; el 29%, psiquiátricos; y el 85%, alteración hepática. En el laboratorio, el 85% presentó ceruloplasmina baja; 55%, cobre urinario alto; en casos con biopsia hepática, 7 tenían depósito de cobre en coloraciones especiales. En neuroimágenes, el 84% presentó hallazgos sugestivos de enfermedad de Wilson y en 3 casos se documentó una mutación genética patogénica. Durante el seguimiento, el 51% mejoró clínica o bioquímicamente, el 11% se mantuvo estable y el 18% se deterioró. El 88% de los casos sobrevivió al final del seguimiento. Conclusiones: este estudio es la cohorte retrospectiva más grande de Colombia. Los resultados son base para nuevos estudios poblacionales buscando de manera activa la enfermedad para documentarla en su fase preclínica y, de este modo, impactar en el pronóstico.

Abstract Introduction: Wilson's disease is a heterogeneous disorder caused by mutations in the ATP7B gene. Its clinical presentation is variable in hepatic and neuropsychiatric phenotypes. The aim of this study is to describe a retrospective cohort of patients. Materials and methods: A descriptive retrospective study was carried out in patients treated at the Hospital Pablo Tobón Uribe from January 2004 to September 2017. Results: 27 patients were reported, 17 men and 10 women. The mean follow-up time was 2.18 years. 40% of the patients had neurological symptoms, 29% psychiatric symptoms, and 85% hepatic impairment. Lab tests showed that 85% had low ceruloplasmin and 55% had increased urinary copper. In cases that underwent liver biopsy, 7 had special copper colorations. Neuroimaging revealed that 84% had findings suggestive of Wilson's disease and a pathogenic genetic mutation was documented in 3 cases. During follow-up, 51% improved clinically or biochemically, 11% remained stable, and 18% deteriorated. 88% of cases survived at the end of follow-up. Conclusions: This study is the largest retrospective cohort carried out in Colombia. The results are the basis for new population-based studies actively seeking this disease to describe its preclinical development and thus impact prognosis.

Mutational analysis of ATP7B gene of hepatolenticular degeneration in Xinjiang region / 中华肝脏病杂志

To understand the mutational characteristics of ATP7B gene of hepatolenticular degeneration in Xinjiang region. 24 cases were diagnosed as hepatolenticular degeneration and the exon of ATP7B gene was detected in some of their siblings and parents. A total of 45 ATP7B gene mutations (93.75%) were detected in 24 cases, of which 14 cases were homozygous mutations or compound heterozygous mutations, six cases were heterozygous mutations and four cases were no mutations. A total of 24 gene mutations and 14 SNPS were detected, including 8 new mutations: c.251C > A, c.121A > c, c.2945C > A, c.2194C > T, c.2947T > c, c.3626T > A, c.3662_3664del, c.3557G > T. The most common mutations were c.2621C > T (p.A874V) [16.7% (4/24)] and c.2333G > T (p.R778L) [12.5% ​​(3/24)]. A total of 4 cases were diagnosed as pre-symptomatic. In this study, the most common mutation in the ATP7B gene is A874V. The most common genetic mutations in Han and Uyghur patients were different. The most common mutation in Han and Uyghur patients is R778L and A874V. Exon 11 is the gene mutations hot spot for patients with hepatolenticular degeneration in Xinjiang region, and is one of the priority exons to be detected when screening patients with suspected hepatolenticular degeneration.

Identification of ATP7B gene variant by combined use of Sanger sequencing, array CGH and quantitative PCR / 中华医学遗传学杂志

Objective@#To identify the type and origin of ATP7B gene mutation in a family affected with Wilson disease by combined use of multiple methods.@*Methods@#Peripheral blood samples were collected from the proband, her parents and her brother. Sanger sequencing were used to detect point mutation and small deletion/insertion of the 21 exons and flanking sequences of the ATP7B gene in all family members. Array-based comparative genomic hybridization (aCGH) was performed to identify copy number variations (CNVs) of the ATP7B gene in the proband. The result was validated by quantitative PCR (qPCR) in other 3 members.@*Results@#Sanger sequencing indicated that the proband carried a heterozygous variation c. 2668G>A (p.V890M) derived from her mother. In addition, 5 common SNPs were detected in her mother, three of which were also identified in her father and brother. The 5 SNPs in the proband were of the wide type. aCGH analysis demonstrated that the proband was heterozygous for a 4 kb deletion, which encompassed exons 2 and 3 of the ATP7B gene and 2 SNPs. qPCR showed that the copy number in her father and brother was about half of the control, indicating heterozygous loss of exons 2 and 3.@*Conclusion@#The combined Sanger sequencing, array CGH and qPCR has identified a novel CNV involving the ATP7B gene. The strategy can improve the diagnostic rate for hereditary or rare diseases.

Análisis de los polimorfismos p. K832R y p. T991T en pacientes cubanos con diagnóstico clínico de la enfermedad de Wilson / p. K832R and p. T991T polymorphisms’ analysis in Cuban patients with clinical diagnosis of Wilson's disease

Introducción: La Enfermedad de Wilson se caracteriza por la acumulación de cobre en hígado, cerebro, riñones y cornea. Se transmite con un patrón de herencia autosómico recesivo. La causa molecular que la provoca son las mutaciones en el gen ATP7B. Se han informado en la literatura más de 139 polimorfismos en el gen ATP7B. Objetivo: Identificar los cambios conformacionales en los exones 10 y 13 y detectar los polimorfismos p.K832R y p.T991T en el gen ATP7B en pacientes cubanos con diagnóstico clínico de Enfermedad de Wilson. Material y Métodos: Se realizó un estudio descriptivo, durante el período 2012 al 2013, que incluyó 27 pacientes con diagnóstico clínico de Enfermedad de Wilson. Para la amplificación del fragmento de interés, se utilizó la técnica de Reacción en Cadena de la Polimerasa y para identificar los cambios conformacionales se aplicó la técnica de Polimorfismo Conformacional de Simple Cadena, en el exón 10 y 13 del gen ATP7B. La presencia de los polimorfismos p.K832R y p.T991T fueron identificados por secuenciación. Resultados: Se detectaron tres cambios conformacionales diferentes denominados: (a, b y c) en el exón 10 y (a y b) en el exón 13 del gen ATP7B. La frecuencia alélica de los polimorfismos p. K832R y p.T991T en 27 pacientes cubanos con diagnóstico clínico de la Enfermedad de Wilson es 35,2 por ciento y 5,6 por ciento respectivamente. Conclusiones: Se analizó por primera vez en Cuba la combinación de los polimorfismos p. K832R y p. T991T que posibilitará hacer estudios moleculares por métodos indirectos(AU)

Introduction: Wilson's disease is characterized by accumulation of copper in the liver, brain and cornea. It is transmitted with an autosomal recessive inherited disorder. The molecular causes are mutations in the ATP7B gene. It has been reported in the literature more than 139polymorphisms of the ATP7B gene. Objective: Identify the conformational changes in exons 10 and 13 and detect the polymorphisms p.K832R and p.T991T in the ATP7B gene in Cuban patients with clinical diagnosis of Wilson's disease. Material and Methods: Was performed a descriptive study including 27 patients with Wilson’s disease ranging in the time from 2012 to 2013. Were applied the polymerase chain reaction to amplify the fragment of interest and the Conformation Polymorphism Single-Chain procedures in the exon 10 and 13 of the ATP7B gene. The p. K832R and p. T991T polymorphisms were detected by sequencing this fragment. Results: Three different conformational changes were identified: (a, b and c) in exon 10 and (a and b) in exon 13 of the ATP7B gene. The allelic frequency of polymorphisms p. K832R and p. T991T in 27 Cuban patients with clinical diagnosis of Wilson's disease is 35.2 percent and 5.6 percent, respectively. Conclusions: It is the first time in Cuba that a combination of the polymorphisms p. K832R and p. T991T were identified which will allow to make possible molecular studies by indirect methods(AU)

Neuro Wilson’s – An Alien Presentation.

Wilson’s disease is an Autosomal recessive disorder of inborn error of copper metabolism in liver which results in accumulation of copper in the liver, brain, kidneys, eye and other organs affecting commonly children and young adults. Its incidence varied from 33 to 68 per 100,000 in India and 1 in 30,000– 40,000 in worldwide population. Mainstay of diagnosis primarily depends on clinical features, biochemical parameters, presence of Kayser-Fleischer (KF) ring. A middle aged man referred as spinocerebellar ataxia was incidentally found to be an exclusive case of Neuro Wilson’s without involvement of the liver and hence we intend to report this case for its rarity.

Clinical analysis of hepatolenticular degeneration in 38 children / 临床儿科杂志

Objective To explore the clinical characteristics and diagnosis of hepatolenticular degeneration (WD) in children. Method The clinical data of 38 children with WD were analyzed retrospectively. Results In the 38 cases (15 males and 23 females), the median age at diagnosis was 6 years, and the average interval between onset and confirmed diagnosis was 5.7 months. The median course of disease was 2 months and the longest was 3 years. Hepatic dysfunction was the most common initial symptom (71.1%), and 27 cases had glutamic acid aminotransferase > 2 ULT (71.1%); Serum ceruloplasmin decreased obviously in 3 cases (94.7%), copper oxidase was significantly reduced in 37 cases (97.4%); 24 h urine copper increased in 33 cases, in which 32 cases(84.2%)had>150 μg/24 h.The K-F rings were presented in 10 cases(26.3%).ATP7B gene sequencing was performed in 19 cases, and the positive rate was 83.3%. Conclusions Onset with liver lesions was common in children with WD, The combination of the results of serum ceruloplasmin, copper oxidase, and 24 h urine copper may made a clinical diagnosis.For a highly suspected case with inadequate evidence,the ATP7B gene detected is helpful.

Clinical analysis of hepatolenticular degeneration in 38 children / 临床儿科杂志

Objective To explore the clinical characteristics and diagnosis of hepatolenticular degeneration (WD) in children. Method The clinical data of 38 children with WD were analyzed retrospectively. Results In the 38 cases (15 males and 23 females), the median age at diagnosis was 6 years, and the average interval between onset and confirmed diagnosis was 5.7 months. The median course of disease was 2 months and the longest was 3 years. Hepatic dysfunction was the most common initial symptom (71.1%), and 27 cases had glutamic acid aminotransferase > 2 ULT (71.1%); Serum ceruloplasmin decreased obviously in 3 cases (94.7%), copper oxidase was significantly reduced in 37 cases (97.4%); 24 h urine copper increased in 33 cases, in which 32 cases(84.2%)had>150 μg/24 h.The K-F rings were presented in 10 cases(26.3%).ATP7B gene sequencing was performed in 19 cases, and the positive rate was 83.3%. Conclusions Onset with liver lesions was common in children with WD, The combination of the results of serum ceruloplasmin, copper oxidase, and 24 h urine copper may made a clinical diagnosis.For a highly suspected case with inadequate evidence,the ATP7B gene detected is helpful.

Enfermedad de Wilson. Actualidad del tema / Wilson’s disease. Theme update

La enfermedad de Wilson constituye un problema de salud mundial. Es una enfermedad hereditaria que si no se atiende de forma adecuada, puede provocar lesiones irreversibles en el hígado y el cerebro, que pueden llevar a la muerte del paciente. En este trabajo se realizó una amplia revisión bibliográfica en bases de datos de la red de Infomed como: LILACS, Hinari, Scielo, Medline/PubMed y en buscadores como: google. Se analizaron 24 artículos referentes a la temática y más del 60% correspondían a los últimos cinco años. La enfermedad de Wilson es poco conocida y la prevalencia mundial es 1:35 000. El diagnóstico clínico se realiza utilizando varias pruebas bioquímicas. La causa molecular que la provoca son las mutaciones en el gen atp7b y en la actualidad se han informado más de 500. En este trabajo se da a conocer complejidad del diagnóstico clínico-molecular de la enfermedad de Wilson.

Wilson’s disease is a world health problem. It is a hereditary disease that could cause irreversible lesions in the liver and brain and lead to patient’s death if it is not adequately treated. In this paper its presents a comprehensive literature review was consulted different databases network in Infomed as: LILACS, Hinari, Scielo, Medline/ PubMed and search engine such as Google. More than 60% correspond to the last 5 years. The prevalence of Wilson’s disease is 1:35000. The clinical diagnosis is achieved using several biochemical tests. The molecular causes that provoking this disease are the mutations in the atp7b gene, and there have been informed more than 500. The difficulty of the clinical and molecular diagnosis was explained in this work.

Mutation Screening of Modifying Genes (Atox1 and Commd1) in Association with Wilson Disease.

Wilson disease (WD) results from accumulation of copper and is caused due to mutations in ATP7B, a copper transporting ATPase. Although WD is an established monogenic disorder, heterogeneity in phenotype is observed even among patients harboring mutations in ATP7B that would affect the mutant protein similarly. Such observations led to the speculation that there might be modifying loci that modulate the phenotype resulting from the aberration in the ATP7B gene. The expected genes coding for proteins that interact either directly with ATP7B or influence it indirectly might fit the role of modifier locus. ATOX1 and COMMD1 are the candidate genes that might play the role of a modifier locus having copper homeostasis pathway with such potential. To understand the role of modifying genes, we screened ATOX1 and COMMD1, a gene implicated in canine copper toxicosis, in 45 WD patients along with 50 healthy controls. This study did not yield satisfactory results concluding more patients to be analyzed. Keywords: Wilson Disease, ATOX1, COMMD1.

Identificación del polimorfismo k832r en pacientes cubanos con diagnóstico clínico de la enfermedad de Wilson / Identification of the k832r polymorphism in patients with clinical diagnosis of Wilson's disease

Introducción: la enfermedad de Wilson se caracteriza por la acumulación de cobre fundamentalmente en el hígado. Se transmite con un patrón de herencia autosómico recesivo. La causa molecular que la provoca son las mutaciones en el gen atp7b. Se han informado en la literatura varios polimorfismos en el gen atp7b. Objetivo: identificar el polimorfismo K832R en 100 pacientes cubanos diagnosticados clínicamente con la Enfermedad de Wilson. Material y Métodos: en el presente estudio se empleó la técnica de cribaje: Polimorfismo Conformacional de Simple Cadena para la determinación de cambios conformacionales en el exón 10. Se utilizó la Técnica de Secuenciación para la identificación del polimorfismo K832R. Resultados: se detectaron tres cambios conformacionales diferentes denominados: a, b y c. El cambio conformacional b y c correspondió al polimorfismo K832R en estado heterocigótico y homocigótico respectivamente. La frecuencia alélica del polimorfismo K832R en 100 pacientes cubanos diagnosticados clínicamente con la Enfermedad de Wilson es de 35%. Conclusiones: se identificó por primera vez en Cuba el polimorfismo K832R y posibilitará hacer estudios moleculares por métodos indirectos.

ABSTRACT Introduction: Wilson's disease is characterized by accumulation of copper in liver, brain and cornea. It is an autosomal recessive inherited disorder of copper metabolism. The molecular causes are mutations in the atp7b gene. It has been reported in the literature several polymorphisms in the atp7b gene. Objective: this research aims to identify the polymorphism K832R in 100 Cubans patients with clinical diagnosis of Wilson's disease. Materials and Methods: in this study we used the technique of screening: single stranded conformational polymorphism for the determination of conformational shifts in exon 10. We used sequencing technique for identifying the K832R polymorphism. Results: they identified three different conformational shifts denominated: a, b and c. The shifts b and c corresponded to polymorphism K832R in heterozygous and homozygous state respectively. The frequency of this polymorphism K832R is 35% in 100 Cubans patients. Conclusions: the polymorphism K832R was identified first in Cuba and it will make possible molecular studies by indirect methods.

Diagnosis of Wilson Disease in Young Children: Molecular Genetic Testing and a Paradigm Shift from the Laboratory Diagnosis / 대한소아소화기영양학회지

Wilson disease (WD) is an autosomal recessive disorder of copper metabolism that results in accumulation of copper primarily in the liver, brain and cornea. Mutations in the WD gene, ATP7B, cause failure of copper excretion from hepatocyte into bile and a defective synthesis of ceruloplasmin. More than 500 mutations are now recognized, scattered throughout the ATP7B gene. Since WD has protean clinical presentations, awareness of WD in clinical practice is important for the early diagnosis and prevention of accumulated copper toxicity. Molecular genetic testing is playing an increasingly important role in the diagnosis of WD in uncertain cases and family screening. Siblings should be screened for WD once an index case has been diagnosed. Discrimination of heterozygotes from asymptomatic patients is essential to avoid inappropriate lifelong therapy for heterozygotes. Genetic testing, either by haplotype analysis or by mutation analysis, is the only definite solution for differentiating heterozygote carriers from affected asymptomatic patients. Routine genetic testing, because of the multitude of documented mutations, has been thought to be impractical until recently. However, genetic testing is now being more actively applied to the diagnosis of WD, particularly in young children in whom conventional biochemical diagnosis has much limitation and only genetic testing is able to confirm WD. Because advancement of modern biochemical technology now allows more rapid, easier, and less expensive mutation detection, direct DNA sequencing could be actively considered as the primary mode of diagnostic investigation rather than a supplementary test to the conventional biochemical tests. This review will focus on the recent advancement of molecular genetics and genetic diagnosis of WD in very young children on the basis of research data of the Seoul National University Children's Hospital and recent literature.

Análisis molecular del exón 2 del gen atp7b en pacientes cubanos con la enfermedad de Wilson / Molecular analysis in the exon 2 of atp7b gene in cubans patients with wilson disease

La enfermedad de Wilson es un trastorno hereditario que se transmite con un patrón de herencia autosómico recesivo. Se caracteriza por la acumulación de cobre fundamentalmente en hígado y cerebro. La causa molecular que la provoca son las mutaciones en el gen atp7b y hasta la fecha se han reportado más de 380. El diagnóstico molecular es complejo. En el presente estudio se empleó la técnica de cribaje: Polimorfismo Conformacional de Simple Cadena para la determinación de cambios conformacionales en el exón 2 en el fragmento a. Se detectó dos cambios conformacionales diferentes denominados: a y b. El cambio conformacional b correspondió a la mutación N41S en estado heterocigótico. La frecuencia alélica de la mutación N41S en 130 pacientes cubanos diagnosticados clínicamente con la enfermedad de Wilson es de un 0.77 por ciento.

Wilson disease is an autosomal recessive inherited disorder of copper metabolism. It is clinically characterised by hepatic and neurological manifestations related to the accumulation of copper in the liver and brain. Molecular analysis reveals more than 380 distinct mutations. The molecular diagnosis is complex. In this investigation we use single- strand conformation polymorphism for determine conformationals shift. We identified two different conformationals shifts in the exon 2 of atp7b gene in cubans patients, denominated: a and b. The shift b correspond with the mutation N41S. The frequency of this mutation is 0.77 percent in 130 cubans patients.

Association of a c.1084A>G (p.Thr362Ala)Variant in the DCTN4 Gene with Wilson Disease

PURPOSE: Wilson disease is an autosomal recessive disorder which causes excessive copper accumulation in the hepatic region. So far, ATP7B gene is the only disease-causing gene of Wilson disease known to date. However, ATP7B mutations have not been found in ~15% of the patients. This study was performed to identify any causative gene in Wilson disease patients without an ATP7B mutation in either allele. MATERIALS AND METHODS: The sequence of the coding regions and exon-intron boundaries of the five ATP7B-interacting genes, ATOX1, COMMD1, GLRX, DCTN4, and ZBTB16, were analyzed in the 12 patients with Wilson disease. RESULTS: Three nonsynonymous variants including c.1084A>G (p.Thr362Ala) in the exon 12 of the DCTN4 gene were identified in the patients examined. Among these, only p.Thr362Ala was predicted as possibly damaging protein function by in silico analysis. Examination of allele frequency of c.1084A>G (p.Thr362Ala) variant in the 176 patients with Wilson disease and in the 414 normal subjects revealed that the variant was more prevalent in the Wilson disease patients (odds ratio [OR]=3.14, 95% confidence interval=1.36-7.22, P=0.0094). CONCLUSION: Our result suggests that c.1084A>G (p.Thr362Ala) in the ATP7B-interacting DCTN4 gene may be associated with the pathogenesis of Wilson disease.

Mutations of ATP7B gene in two Thai siblings with Wilson disease

Background: Wilson disease (WD) is an autosomal recessive disorder of copper metabolism caused by mutations in ATP7B gene. Objective: Report the clinical data and mutation analysis of two Thai siblings suspected of WD. Subject and methods: A 13-year-old boy who presented with cirrhosis, arthralgia, hypoalbuminemia, and coagulopathy, and his 11-year-old sister who was asymptomatic but had hepatomegaly with elevation of transaminases, were studied. Mutation analysis of ATP7B gene and mRNA analysis was performed in both patients and their parents. Results: Investigations were consistent with WD, and their liver diseases improved after standard treatment for WD. DNA analyses in these two patients revealed two novel mutations, which were a deletion of the first 2bp of exon 6 (c.1870_1871delGA), and a single base substitution from A to G at nucleotide 4075 (c.4075A>G) in the exon 20 (p.M1359V). PCR-restriction digestion with NcoI restriction enzyme was employed as the second method for confirmation of the c.4075A>G mutation and for rapid screening in 100 chromosomes from unrelated healthy controls, and this variant was not present in the controls. The c.1870_1871delGA deletion caused a frameshift effect, which results in a premature stop codon (p.E624fsX753), and the p.M1359V mutation is a substitution of methionine with valine, which may have effects upon its orientation and interaction with other adjacent amino acids. Conclusion: Two novel mutations of ATP7B gene were identified in two Thai siblings with WD.

Mutations in exon of ATP7B gene in Chinese patients with Wilson's disease / 第二军医大学学报

Objective: To amplify the ATP7B gene of Wilson disease (WD) patients by PCR and to sequence the amplification product, so as to characterize the possible mutations. Methods: The genomic DNA of 41 WD patients, 10 normal controls and a WD genealogy (proband's daughter and parents) were extracted. The fragments of exon 8 and 12 of ATP7B gene were amplified using PCR, and the PCR products were directly sequenced. Results: No abnormality was found in the control group. Mutations of exon 8 were found in 11 WD patients, with 6 WD patients having Arg778Leu heterozygous mutations. Mutations of exon 12 were found in 4 WD patients, with 2 patients having Arg952Lys mutations. In the sibs of the WD patient, the proband's daughter carried 2 heterozygous mutations: Arg778Leu mutation of exon 8 was from her father and Pro992Leu mutation of exon 13 from her mother; the proband's parents were found as normal heterozygous carriers. Conclusion: Exon 8 and 12 of ATP7B gene are prone to mutations in Chinese WD population, and the mutations of other exons, such as exon 13, also exist.

Haplotype analysis at R778L mutation of ATP7B gene in Korean patients with Wilson disease / 대한간학회지

No abstract available.

Haplotype analysis and possible founder effect at the R778L mutation of the ATP7B gene in Korean patients with Wilson's disease / 대한간학회지

BACKGROUND/AIMS: Wilson's disease (WD) is an inherited disorder of copper metabolism caused by alteration of the P-type adenosine triphosphatase (ATP) 7B gene. In this study, we analyzed the frequency of well-known mutations and constructed the first haplotypes for Koreans. In addition, we evaluated whether a founder effect existed in Korean patients with WD. METHODS: We obtained DNA samples from 21 patients with WD and their parents (total cohort n=63). ATP7B gene mutations were identified by direct sequencing methods, and microsatellite typing was performed at D13S315, D13S1325, and D13S316 with fluorescent dye-labeled primers. Any founder effect was identified by using 42 normal alleles from parents with a normal phenotype as a control group. The chi-square test and Fisher's exact test were used for statistical analysis. RESULTS: Three common mutations were found in 23 chromosomes obtained from 21 patients: the R778L mutation at exon 8 (15/23, 65.2%), the A874V mutation at exon 11 (6/23, 26.1%), and the N1270S mutation at exon 18 (2/23, 8.7%). D13S315 and D13S316 showed linkage disequilibrium at alleles 5 and 4, respectively, in patients with the R778L mutation (P=0.0157 and 0.0001, respectively). The haplotype made up of these two alleles occurred significantly more frequently in patients with the R778L mutation (5-R778L-4, D13S315-mutation-D13S316) than in the controls (P=0.0018). CONCLUSIONS: The arche haplotype of the ATP7B gene in Korean patients with WD may be 5-R778L-4 (D13S315.mutation.D13S316), and it might illustrate a founder effect.

Establishment and characterization of secondary drug resistant cell line originating from cisplatin-resistant SNU-8 / 대한산부인과학회지

OBJECTIVE: Multidrug resistance to chemotherapy is a major obstacle in attempts to improve the clinical outcome of ovarian carcinoma patients. The aim of this study is to establish secondary anticancer drug resistant cell line from original SNU-8/WT resistant to cisplatin and characterize it. METHODS: After establishing secondary drug resistant cell line (SNU-8/Fac), drug sensitivity was measured by MTT assay. Multidrug resistance (MDR) was analyzed by RT-PCR and western blotting analysis. RESULTS: MTT assay data demonstrated that MDR was expressed in SNU-8/Fac. In addition, mRNA expression of MDR1, ATP7B in SNU-8/Fac was increased. However, overexpression of MRP1, BCRP, TS and MT mRNA was not observed. At the protein level, protein P-gp, ATP7B were overexpressed in SNU-8/Fac. CONCLUSION: We established a new anticancer drug resistant cell line from original SNU-8/WT. Overexpression of P-gp and ATP7B was observed in SNU-8/Fac.

Study on the Diagnosis Technology of Wilson's Disease Gene Mutation with Double PCR-SSCP / 医学研究杂志

Objective To establish a fast and effective gene diagnosis method for Wilson's disease(WD) patients by double PCR-SSCP technology.Methods We amplificated exon8 and exon12 of ATP7B gene by double PCR from genomic DNA of 140 unrelated WD patients and 30 normal controls,then used SSCP technology to screen them.At last we identificated these patients' mutation features by direct sequencing.Results No abnormality shift was found in 30 controls.In 140 patients,we found 7 types of abnormal mobility shifts in 66 cases(47.14%).In subsequent direct sequencing,mutation rate of Arg778Leu/Gln was 37.14%(52/140),and Thr935Met was 12.86%(18/140).Conclusion Double PCR-SSCP diagnosis technology is a effective method which can improve diagnosis rate for Wilson disease.