Hereditary hemorrhagic telangiectasia complicated by aortic sinus aneurysm caused by a pathogenic mutation in the ACVRL1 gene: a family survey / 中华皮肤科杂志

Objective:To investigate clinical and genetic characteristics of a family with hereditary hemorrhagic telangiectasia complicated by aortic sinus aneurysm, and to analyze causative genes.Methods:Clinical data and peripheral blood samples were collected from the proband and her relatives, and genomic DNA was extracted. Causative genes were screened by whole-exome sequencing, and then verified by Sanger sequencing.Results:A heterozygous mutation c.137G>A was identified at position 137 in exon 3 of the ACVRL1 gene in the proband, her daughter, grandson and granddaughter, which led to the substitution of cysteine by tyrosine at amino acid position 46 (p.C46Y) . The mutation was not found in any of the other 5 family members without clinical symptoms.Conclusion:A causative mutation c.137G>A (p.C46Y) in the ACVRL1 gene was identified in the family with hereditary hemorrhagic telangiectasia type 2 complicated by aortic sinus aneurysm, which had not been previously reported in Asian populations.

Newborn screening, gene variation analysis and follow-up study of very long chain acyl CoA dehydrogenase deficiency / 中华实用儿科临床杂志

Objective:To investigate the prevalence, gene variation and prognosis of very long chain acyl CoA dehydrogenase deficiency (VLCADD) in newborns in Henan Province.Methods:From January 2013 to December 2019, 867 103 newborns were investigated for VLCADD by tandem mass spectrometry.Children who diagnosed as VLCADD and their families were subjected to next-generation sequencing and Sanger sequencing.Clinical data, biochemical changes and gene variation characteristics of the confirmed cases of VLCADD were analyzed.Dietary guidance was given, and their growth and development were followed up.Results:Six neonates were diagnosed as VLCADD, and the prevalence of VLCADD in the Henan Province was 1/144 517.A total of 11 mutations in the ACADVL gene were found, including 5 new variants c. 692-2_692-1delAG, c.753-23_753-22del, c.960delG, c.1361A>G, and c. 1955C>T.The newborns were given a high-carbohydrate, low-fat diet, and followed up for 8-56 months.Except for two deaths, all patients had a good outcome. Conclusions:The prevalence of neonatal VLCADD in Henan Province is 1/144 517.This results has enriched the ACADVL gene mutation spectrum and provided an important basis for the screening and diagnosis of VLCADD.

Spatiotemporal distribution of oncomelania hupensis snail habitats in the Middle and Lower Reaches of the Yangtze River / 公共卫生与预防医学

Schistosomiasis is one of the major communicable diseases, which seriously impacts human health, Oncomelania hupensis snail is unique intermediate host of Schistosoma japonicum as well as posts significant influence on schistosomiasis transmission. The long-term serial data of oncomelania snail area in the Middle and Lower Reaches of the Yangtze River in the Schistosomiasis situation in People's Republic of China ,were collected from 1999 to 2018. The spatial and temporal variation characteristics of oncomelania hupensis area in five provinces were analyzed by Manner-Kendall test. In the spatial change, the area composition ratio of lake-and-marshland regions in Hunan increased, while that in Hubei and Jiangxi showed a slight decline. The lake-and-marshland snails in Anhui showed a significant downward trend, while Jiangsu showed no stable trend. In the spatial change, the area of lake-and-marshland snails in Jiangsu, Anhui, and Hubei showed a downward trend, among which the downward trend in Hubei was not obvious, while the upward trend in Jiangxi and Hunan had an upward trend but the trend was not obvious. the year of the sudden change in the area of the lake and marshes in in Jiangsu and Hubei were 2010 and 2017 ; There was no significant mutation in Anhui Province; the mutation point of the lake-and-marshland snails in Hunan was between 2001 and 2002; the mutation year of the lake-and-marshland snails in Jiangxi was 2002.This paper studies the temporal and spatial changes of the snail breeding area in the middle and lower Yangtze River, and the results can provide a basis for the elimination of snails in different types of distribution areas.

Spectrum analysis of gene mutations in retinitis pigmentosa and cone-rod dystrophy / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To analyze the gene mutation spectrum of autosomal recessive retinitis pigmentosa(ARRP)pedigrees and cone-rod dystrophy(CORD)pedigrees in Ningxia region of China. <p>METHODS:Totally 35 ARRP pedigrees and 18 CORD pedigrees were included in Ningxia Eye Hospital from September 2016 to February 2020. Peripheral venous blood samples of the proband were collected for targeted capture enrichment and high-throughput sequencing using a genetic retinal disease capture chip that contain 232 pathogenic genes. Online analysis software was used to predict the pathogenicity of suspicious gene variation, and Sanger sequencing was used to analyze the co-segregation of the family members. <p>RESULTS: Totally 16 pathogenic genes were confirmed in 35 ARRP pedigrees, the mutations rate of RP1 gene was the highest, accounting for 14%(5/35), following were ABCA4, CRB1 and EYS gene, accounted for 11%(4/35); 18 CORD pedigrees carried 10 pathogenic genes. The mutation rate of ABCA4 gene was the highest, accounting for 28%(5/18), followed by ALMS1, PROM1, RPE65, USH2A gene, accounting for 11%(2/18). There were 5 co-exist disease-causing genes in ARRP and CORD pedigrees, which were ABCA4, CLN3, CRB1, PROM1, NRL, accounting for 42%(22/53).<p>CONCLUSION: There are similarities and crossover in the phenotype of ARRP and CORD. The pathogenic genes were overlaped. The most common overlaping gene between the two diseases is ABCA4.

H396P mutation in chronic myeloid leukaemia patient on nilotinib – A case report

@#Introduction: The advent of BCR-ABL1-targeted therapy with the tyrosine kinase inhibitor (TKI), for example, imatinib and nilotinib, marked a turning point in the therapy of chronic myeloid leukaemia (CML). However, a substantial proportion of patients experience primary or secondary disease resistance to TKI. There are multifactorial causes contributing to the treatment failure of which BCR-ABL1 kinase domain mutation being the most common. Here, we describe a case of a CML patient with H396P mutation following treatment with nilotinib. Case: A 60-year-old woman presented with abdominal discomfort and hyperleukocytosis. She was diagnosed as CML in the chronic phase with positive BCR-ABL1 transcripts. Due to the failure to obtain an optimal response with imatinib treatment, it was switched to nilotinib. She responded well to nilotinib initially and achieved complete haematological and cytogenetic responses, with undetectable BCR-ABL1 transcripts. However, in 4 years she developed molecular relapse. Mutation analysis which was done 70 months after commencement of nilotinib showed the presence of BCRABL1 kinase domain mutation with nucleotide substitution at position 1187 from Histidine(H) to Proline(P) (H396P). Currently, she is on nilotinib 400mg twice daily. Her latest molecular analysis showed the presence of residual BCR-ABL1 transcripts at 0.22 %. Discussion/conclusion: This case illustrates the importance of BCR-ABL1 mutation analysis in CML patients with persistent BCR-ABL1 positivity in spite of treatment. Early detection and identification of the type of BCRABL1 mutation are important to guide appropriate treatment options as different mutation will have different sensitivity to TKI.

Genetic Mutation Screening of DNA Polymerase in Human Lung Cancer / 中国肺癌杂志

BACKGROUND@#DNA polymerase β is one of the key enzymes for DNA repair and it was reported that about 30 percent of different types of cancers carried mutations in its coding gene Polb. However, it is still controversial whether it is true or false because of the small sample size in these studies. In current study, we performed genetic screening of promoter and coding regions of Polb gene in 69 Chinese lung cancer patients using Sanger sequencing method, so as to elucidate real mutation frequency of Polb mutations in Chinese Han population.@*METHODS@#Salting out extraction method was used to get the genome DNAs from tumor and normal matched tissues of 69 lung cancer patients. The promoter and 14 coding regions of Polb gene were then amplified using these DNAs as the template. After purification, amplicons were sequenced and aligned to the wild type Polb gene in NCBI database, in order to find out the mutated sites of Polb gene in Chinese lung cancer patients.@*RESULTS@#In this study, we totally found only 5 mutated sites in Polb gene. In detail, 3 mutations (-196G>T, -188_-187insCGCCC, -168C>A) were located in the promoter region; 2 mutations (587C>G, 612A>T) were found in coding regions. Specially, mutations of -188_-187insCGCCC and 587C>G (resulting to the amino acid substitution of Thr to Ser at position 196) had never been reported by other groups before. However, all these 5 mutated sites could be detected in both tumor and matched normal tissues, which inferred that they are not lung tumor specific mutations.@*CONCLUSIONS@#No lung tumor specific mutations of Polb gene could be found in Chinese lung cancer patients and Polb gene mutation might not be a molecular marker for Chinese lung cancer patients.

Mutation of MED12 is not a frequent occurrence in prostate cancer of Korean patients / 亚洲男科学杂志(英文版)

Prostate cancer is one of the major health care problems, but the molecular pathogenesis has been relatively insufficiently elucidated. Recently, whole exome sequencing of prostate cancer identified recurrent mutations involving MED12 in Caucasian patients, which finding was not reproduced in one subsequent study by Sanger sequencing. Thus, we investigated mutation status of MED12 in exons 2 and 26 by Sanger sequencing in 102 radical prostatectomy cases from Korean patients. The analysis found the mutation in none of the cases. Therefore, MED12 mutation does not appear to represent a significant molecular alteration in this cohort of patients according to the analysis by the traditional gold standard.

BRAF, KIT, NRAS, GNAQ and GNA11 mutation analysis in cutaneous melanomas in Turkish population.

Background: KIT and mitogen-activated protein kinase cascade are important for melanomagenesis. In the present study, we analyzed the frequency of BRAF, NRAS, KIT, GNAQ and GNA11 gene mutations and investigated their association with clinicopathological features of melanomas in Turkish population. Materials and Methods: Forty-seven primary cutaneous melanomas were included in our study. Sanger sequencing method was used for mutation analysis in all cases. Results: Mean age was 62.1 (29-101) years. Female:male ratio was 17:30. Among 47 melanomas, 14 (29.8%) BRAF, 10 (21.3%) NRAS, 4 (8.5%) KIT and 1(2.1%) GNAQ gene mutations were detected. Two of the KIT mutations were found in acral lentiginous melanoma (ALM). In the head and neck region, mutation frequency was signifi cantly lower than in other locations (P = 0.035). The only GNAQ gene mutation (p.Q209L) was detected in a melanoma arising from blue nevus located on the scalp. None of the melanomas harbored NRAS exon 2, KIT exon 13/17/18, GNAQ exon 4 and GNA11 exon 4/5 mutations. Overall mutation frequency did not show signifi cant difference between metastatic (8/14, 57.1%) and nonmetastatic (18/33, 54.5%) patients. We did not observe any signifi cant association between mutation status and gender or age of various patients. Conclusions: Our results support that BRAF and NRAS gene mutations are common in cutaneous melanomas. The activating mutations of KIT gene are rare and especially seen in ALM. GNAQ and GNA11 mutations are infrequent in cutaneous melanomas and may be associated only with melanomas arising from blue nevus.

Hepatitis B virus (HBV) gene mutations related to entecavir-resistance in patients with chronic HBV infections / 中华临床感染病杂志

Objective To investigate hepatitis B virus (HBV) gene mutations related to entecavir (ETV)-resistance in patients with chronic HBV infections.Methods Serum samples were collected from 44 patients with chronic HBV infections and resistant to ETV treatment who were admitted in Ningbo No.2 Hospital during February 2010 and May 2014.The HBV polymerase regions were amplified by real-time fluorescent quantitative polymerase chain reaction (PCR) method,and the PCR products were analyzed with direct sequencing.SPSS 16.0 was used to assess the frequency of HBV polymerase gene mutations,and its relation to the viral genotype and clinical features.Results The most common HBV polymerase gene mutation was rtS202G/I (52.28％,23/44),followed by rtT184A/G/I/S (36.36％,16/44) and rtM250V/L (11.36％,5/44).Nine mutation patterns were detected,in which rtL180 + rtM204V + rtS202G/I (38.64％,17/44) and rtL180 +rtM204V + rtT184A/G/I/S (27.27％,12/44) were the most frequent ones.The difference in gene mutations between genotype B and C was of statistical significance (x2=12.294,P ＜0.01).Patients carrying rtT184A/G/I/S mutations were associated with worse liver function (x2 =14.499,P ＜ 0.01),and those carrying rtM250V/L mutations were associated with lower HBeAg positive rate (x2 =10.057,P ＜ 0.01).Conclusions rtL180M + rtM204V + rtS202G/I is the most common HBV polymerase gene mutation related to ETV resistance in patients with chronic HBV infections.Different gene mutations may be associated with HBV genotypes,severity of liver damages,and HBeAg positive rate.

Spectrum of K ras mutations in Pakistani colorectal cancer patients

The incidence of colorectal cancer (CRC) is increasing daily worldwide. Although different aspects of CRC have been studied in other parts of the world, relatively little or almost no information is available in Pakistan about different aspects of this disease at the molecular level. The present study was aimed at determining the frequency and prevalence of K ras gene mutations in Pakistani CRC patients. Tissue and blood samples of 150 CRC patients (64% male and 36% female) were used for PCR amplification of K ras and detection of mutations by denaturing gradient gel electrophoresis, restriction fragment length polymorphism analysis, and nucleotide sequencing. The K ras mutation frequency was found to be 13%, and the most prevalent mutations were found at codons 12 and 13. A novel mutation was also found at codon 31. The dominant mutation observed was a G to A transition. Female patients were more susceptible to K ras mutations, and these mutations were predominant in patients with a nonmetastatic stage of CRC. No significant differences in the prevalence of K ras mutations were observed for patient age, gender, or tumor type. It can be inferred from this study that Pakistani CRC patients have a lower frequency of K ras mutations compared to those observed in other parts of the world, and that K ras mutations seemed to be significantly associated with female patients.

Estudo da Beta-catenina em tumores adrenocorticais humanos / Study of beta-catenin in human adrenocortical tumors

Introdução: A incidência de tumores adrenocorticais em crianças é particularmente elevada nas regiões sudeste e sul do Brasil, correlacionandose com a ocorrência da mutação germinativa p.R337H do supressor tumoral p53, entretanto, o carcinoma adrenocortical é uma neoplasia endócrina maligna rara em todo o mundo com uma incidência aproximada de 0,5 - 2 casos por milhão por ano. Esta condição é uma doença heterogênea, apresentando frequentemente comportamento clínico agressivo e letal. A cascata de sinalização Wnt é uma via importante de transdução de sinal em cânceres humanos e tem sido implicada na tumorigênese adrenocortical. A atividade desta via de sinalização é dependente da quantidade de beta-catenina citoplasmática e nuclear. Mutações ativadoras no gene da beta-catenina (CTNNB1) foram relatadas em diversas neoplasias humanas. Estudos demonstraram que mutações no gene CTNNB1 são os defeitos genéticos mais frequentemente encontrados em adenomas e em carcinomas adrenocorticais. O estudo destas mutações demonstrou que as alterações no gene CTNNB1 localizam-se principalmente exon 3, que codifica a porção amino terminal da beta- catenina. Objetivos: determinar a ocorrência e a frequência das mutações somáticas no exon 3 do gene CTNNB1. Adicionalmente, determinar a imunorreatividade de beta-catenina e de p53 em tumores adrenocorticais benignos e malignos de crianças e adultos. Correlacionar os resultados da análise de mutações gênicas e os dados de imunorreatividade com as características hormonais, a mutação p.R337H do p53, o diagnóstico histológico e a evolução dos tumores adrenocorticais de crianças e adultos. Métodos: Neste estudo, a análise de imunohistoquímica para beta-catenina e p53 foi realizada em 103 tumores adrenocorticais benignos e malignos (40 crianças e 63 adultos), estando as amostras histológicas alocadas em micromatriz tecidual (TMA). A pesquisa de mutações no exon 3 do gene CTNNB1 foi determinada por seqüenciamento automático em 64 tumores...

Introduction: The incidence of adrenocortical tumors in children is particularly high in the southeastern and southern regions of Brazil, correlating with the occurrence of p.R337H p53 tumor suppressor germline mutation. However, adrenocortical carcinoma is a worldwide rare endocrine malignancy with an approximate incidence of 0.5 to 2 cases per million per year. This condition is a heterogeneous disease and is often lethal. The Wnt signaling pathway is an important signal transduction pathway in human cancers and has been implicated in adrenocortical tumorigenesis. The activity of this signaling pathway is dependent on the amount of nuclear and cytoplasmic beta-catenin. Activating mutations of ?-catenin (CTNNB1) gene have been reported in several human malignancies. Studies have shown that CTNNB1 mutations are the most common genetic defect found in adrenocortical adenomas and carcinomas. The study of these mutations demonstrated that the changes in CTNNB1 gene are mainly located in exon 3, which encodes the amino terminal portion of the beta- catenin. Objectives: to determine the occurrence and frequency of CTNNB1 somatic mutations and the abnormal beta-catenin and p53 accumulation in benign and malignant adrenocortical tumors in both children and adults. We also evaluated the correlation of the gene mutations analysis and immunohistochemistry data with the hormonal characteristics, the p.R337H germline mutation, the histological diagnosis and the prognosis of adrenocortical tumors in children and adults. Methods: In this study, immunohistochemistry for beta-catenin and p53 was performed in 103 benign and malignant (40 children and 63 adults) adrenocortical tumors. The histological samples were allocated in a tissue microarray (TMA). The study of the CTNNB1 gene was performed by direct sequencing of 64 adrenocortical tumors. Results: The beta-catenin abnormal accumulation was similar in benign and malignant adrenocortical tumors of children and adults (15...(au)

Analysis of gene mutations among South Indian patients with maple syrup urine disease: Identification of four novel mutations.

Maple syrup urine disease (MSUD) is predominantly caused by mutations in the BCKDHA, BCKDHB and DBT genes, which encode for the E1α, E1β and E2 subunits of the branched-chain α-keto acid dehydrogenase complex, respectively. Because disease causing mutations play a major role in the development of the disease, prenatal diagnosis at gestational level may have significance in making decisions by parents. Thus, this study was aimed to screen South Indian MSUD patients for mutations and assess the genotype-phenotype correlation. Thirteen patients diagnosed with MSUD by conventional biochemical screening such as urine analysis by DNPH test, thin layer chromatography for amino acids and blood amino acid quantification by HPLC were selected for mutation analysis. The entire coding regions of the BCKDHA, BCKDHB and DBT genes were analyzed for mutations by PCR-based direct DNA sequencing. BCKDHA and BCKDHB mutations were seen in 43% of the total ten patients, while disease-causing DBT gene mutation was observed only in 14%. Three patients displayed no mutations. Novel mutations were c.130C>T in BCKDHA gene, c. 599C>T and c.121_122delAC in BCKDHB gene and c.190G>A in DBT gene. Notably, patients harbouring these mutations were non-responsive to thiamine supplementation and other treatment regimens and might have a worse prognosis as compared to the patients not having such mutations. Thus, identification of these mutations may have a crucial role in the treatment as well as understanding the molecular mechanisms in MSUD.

Alkaptonuria: A very rare metabolic disorder.

Alkaptonuria (AKU) is a very rare autosomal recessive disorder of tyrosine metabolism in the liver due to deficiency of homogentisate 1,2 dioxygenase (HGD) activity, resulting in the accumulation of homogentisic acid (HGA). Circulating HGA pass into various tissues through-out the body, mainly in cartilage and connective tissues, where its oxidation products polymerize and deposit as a melanin-like pigment. Gram quantities of HGA are excreted in the urine. AKU is a progressive disease and the three main features, according the chronology of appearance, are: darkening of the urine at birth, then ochronosis (blue-dark pigmentation of the connective tissue) clinically visible at around 30 yrs in the ear and eye, and finally a severe ochronotic arthropathy at around 50 yrs with spine and large joints involvements. Cardiovascular and renal complications have been described in numerous case report studies. A treatment now is available in the form of a drug nitisinone, which decreases the production of HGA. The enzymatic defect in AKU is caused by the homozygous or compound heterozygous mutations within the HGD gene. This disease has a very low prevalence (1:100,000-250,000) in most of the ethnic groups, except Slovakia and Dominican Republic, where the incidence has shown increase up to 1:19,000. This review highlights classical and recent findings on this very rare disease.

THE STUDY OF MUTATION SPECTRUM OF HEMOPHILIA / Шинэ санаа Шинэ нээлт

Background:Hemophilia is life-threatening and hereditary bleeding disorder caused by deficiencies of pi coagulation VIII. IX, XI factor, and that is inherited by X-linkcd recessive.However, hemophilic treatment is getting increase in our country, but still insufficient or poor because of high cost, which is about 50000-70000$ per patient per year in other countries. Now, we need to detect hemophilia heterozygote (XAX*) or no symptomatic carrier among hemophiliac siblings (including mother, younger sister, sister) that is essential for prevention of hemophilia. We need to screen the mutation spectrum of FS and F9 gene among patients with hemophilia in order to detect hemophilia carriers.Objective:to detect a mutation 1'Ó ø! f9 gene among patients with hemophilia Methods:The blood samples were collected from paffeflf* who had been hospitalizing in department ot hematology of the First Clinical Hospital and the National Center for Maternal and Child Health from 2010 to 2011. We have carried out the screening of most common mutation named int- 22 inversion by Long-Range PC'R method is previously described by Lui Q et al.. 1998. Direct sequence method was used to detect the SNP and small deletion in patients who had no int-22 inv and large deletion.Results:In total, If pillitfhis with hemophilia (ÈË-14, IIB=1) participated in this study. The 9 patients positive for an int-22 inversion mutation, while the one patient had a multiple exon deletion (exon 1-13) which was demonstrated by repeated PC'R amplification failure. I wo missense mutation and 1 frame shift mutation were detected. The one patient had nonsense mutation but he was diagnosed as a severe hemophilia-A patient.Conclusion:We have to urgently adopt the molecular diagnosis and carrier detection ol hemophilia in our rnuntrv

K-ras gene mutation status in 454 Chinese patients with colorectal cancer / 肿瘤

Objective: To analyze the mutation status of K-ras gene in CRC (colorectal cancer) tissues and plasma samples of CRC patients, ultimately to promote the targeted agents against EGFR (epidermal growth factor receptor) (such as cetuximab and panitumumab, etc.) used in personalized treatment for CRC patients. Methods: The sequence of K-ras gene at specific sites was investigated in 431 CRC tissues and 23 plasma samples collected from 454 CRC patients, respectively. To improve the detection sensitivity, a combinatory approach was chosen which included the COLD-PCR (coamplification at lower denaturation temperature PCR) method, followed by DNA sequencing. Chi-square test was employed to analyze K-ras mutation frequencies in various sample types or subgroups of CRC patients according to age, and the difference was considered statistically significant if P value was less than 0.05. Results: The overall K-ras mutation rate was 25.29% in 431 CRC tissue samples. The two major forms of K-ras mutation were Gl 2D (mutation of glycine to an aspartate residue at codon 12) and Gl 3D (mutation of glycine to an aspartate residue at codon 13), and their occurrence frequencies were 12.99% and 6.26%, respectively. Interestingly, the overall K-ras mutation rate was 21.74% in blood samples from additional 23 CRC patients, without a significant difference from the mutation rate in the tumor samples (P > 0.05). Moreover, the occurrence frequency of anyone form of K-ras mutation was 37.94% in CRC patients aged 60 and over, which was significantly higher than the detection rate (7.30%) of the patients under the age of 60 (P < 0.01). Conclusion: COLD-PCR amplification combined with DNA sequencing method can be used to detect the mutation status of K-ras gene, and plasma samples can be used insteadly if CRC tumor tissues are unavailable. In addition, for elderly patients aged of 60 and over, it is suggested that K-ras mutation status should be routinely detected before treatment. Copyright © 2013 by TUMOR.

THE STUDY OF MUTATION SPECTRUM OF HEMOPHILIA / Шинэ санаа Шинэ нээлт

Background: Hemophilia is life-threatening and hereditary bleeding disorder caused by deficiencies of pi coagulation VIII. IX, XI factor, and that is inherited by X-linkcd recessive. However, hemophilic treatment is getting increase in our country, but still insufficient or poor because of high cost, which is about 50000-70000$ per patient per year in other countries. Now, we need to detect hemophilia heterozygote (XAX*) or no symptomatic carrier among hemophiliac siblings (including mother, younger sister, sister) that is essential for prevention of hemophilia. We need to screen the mutation spectrum of FS and F9 gene among patients with hemophilia in order to detect hemophilia carriers. Objective: to detect a mutation 1'Ó ø! f9 gene among patients with hemophilia Methods: The blood samples were collected from paffeflf* who had been hospitalizing in department ot hematology of the First Clinical Hospital and the National Center for Maternal and Child Health from 2010 to 2011. We have carried out the screening of most common mutation named int- 22 inversion by Long-Range PC'R method is previously described by Lui Q et al.. 1998. Direct sequence method was used to detect the SNP and small deletion in patients who had no int-22 inv and large deletion. Results: In total, If pillitfhis with hemophilia (ÈË-14, IIB=1) participated in this study. The 9 patients positive for an int-22 inversion mutation, while the one patient had a multiple exon deletion (exon 1-13) which was demonstrated by repeated PC'R amplification failure. I wo missense mutation and 1 frame shift mutation were detected. The one patient had nonsense mutation but he was diagnosed as a severe hemophilia-A patient. Conclusion: We have to urgently adopt the molecular diagnosis and carrier detection ol hemophilia in our rnuntrv

Candidate genes prediction in pakistani families with hearing impairment by using bioinformatic approach / Predição de genes candidatos em famílias paquistanesas com deficiência auditiva utilizando uma abordagem de bioinformática

Multiple factors such as genetic and environmental, are involved in causing hearing impairment (HI). Severe or profound hearing loss affects approximately one in 1000 children worldwide and half of these cases are due to genetic factors. In case of hereditary nonsyndromic HI, approximately 75­80% of cases are involved in autosomal recessive inheritance and 15% of cases involve autosomal dominant inheritance. HI represents extreme genetic heterogeneity. In nonsyndromic deafness, 135 loci have been mapped till now including 77 autosomal recessive genes of which only 29 corresponding nuclear genes have been cloned. This study was designed to apply bioinformatic approach for reducing large number of candidate genes responsible for deafness to a handy number for their mutation analysis. Databases of expressed mouse inner ear genes and the expressed human cochlear genes were used to cross-reference all genes present in particular locus predicting candidate genes for phenotypes of nonsyndromic hereditary HI. These candidate genes are a source of starting point for mutation analysis along with genetic linkage to refine the loci. After characterization, it was observed that KIAA119 and EDN3 are candidate genes for deafness. In present study, there were total 14 loci and two genes KIAA119 and EDN3 were identified as candidate genes in locus 48 and locus 65 respectively. If mutation analysis of the two characterized genes is done, it will not be a comparatively time taking and labor-intensive process as these genes are only two in number.

Diversos fatores, tais como genéticos e ambientais, estão envolvidos na causa da deficiência auditiva (HI). A perda auditiva severa ou profunda afeta aproximadamente uma em cada 1000 crianças em todo o mundo e metade destes casos são devidos a fatores genéticos. Em relação a HI não-sindrômica hereditária, cerca de 75-80% dos casos estão envolvidos na herança autossômica recessiva e 15% dos casos envolvem herança autossômica dominante. HI representa extrema heterogeneidade genética. Em casos de surdez, 135 loci foram mapeados até agora, incluindo 77 genes autossômicos recessivos das quais apenas 29 genes correspondentes nucleares foram clonados. Este estudo foi desenhado para aplicar abordagem de bioinformática a fim de reduzir o grande número de genes candidatos responsáveis pela surdez a um número útil para a análise de mutação. Bases de dados de genes expressos do ouvido interno em camundongos e de genes expressos na cóclea em humanos foram usados para cruzar todos os genes presentes no locus específico prevendo genes candidatos para os fenótipos de HI não sindrômica hereditária. Estes genes candidatos são uma fonte de ponto de partida para a análise de mutação, juntamente com a ligação gênica para refinar os locos. Após a caracterização, verificouse que KIAA119 e EDN3 são genes candidatos para a surdez. No presente estudo, houve um total de 14 locos e dois genes KIAA119 e EDN3 foram identificados como genes candidatos no locus 48 e locus 65, respectivamente. Se a análise de mutação dos dois genes caracterizados for feita, não será um processo comparativamente longo e trabalhoso uma vez que são apenas dois genes.

Spectrum of Lysosomal Storage Disorders at a Medical Genetics Center in Northern India.

Background: There is limited literature available on the phenotypic and mutation spectrum of Indian patients with Lysosomal storage disorders (LSD). Objective: To elucidate the clinical, biochemical and mutation spectrum and to study the management options in Indian patients with lysosomal storage disorders. Design: Descriptive study. Subjects and Methods: All patients with lysosomal storage disorders diagnosed in the Medical Genetics department of a tertiary care institute in North India over a three year period from January 2008 to December 2010. Results: Out of the total of 93 patients clinically suspected to have LSDs, 68 (mean age at presentation 4.5 years) were confirmed to have LSDs based on the laboratory/neuroimaging findings and documentation of deficient enzymatic activity in the peripheral blood (leucocytes or plasma) and/or skin fibroblasts. The commonest clinical features at presentation were growth retardation (failure to thrive 47.2% and short stature 17.6%), hepatosplenomegaly (41.2%) and neuroregression (33.8%). A history of consanguinity was present in 32.4% of the families. Prenatal diagnosis was done in a total of 6 affected families; two pregnancies were found to be affected (one each with Gaucher disease and Tay Sachs disease) and in both cases the parents opted for termination of pregnancy. Of the remaining four pregnancies which were found to be unaffected and therefore continued, three were confirmed to be normal on post-natal follow up. Enzyme replacement therapy (ERT) is being given for a total of 8 LSD patients and all of them are showing a gradual amelioration of their symptoms and an improvement in the quality of life. Conclusions: Lysosomal storage disorders constitute an important group of genetic metabolic disorders for many of which therapeutic options are now available.

Detection of epidermal growth factor receptor mutations in small specimens of non-small cell lung cancer by amplification refractory mutation system / 肿瘤

Objective: To explore the feasibility of the application of small specimens as alternatives of gross specimens to detect EGFR (epidermal growth factor receptor) mutation in NSCLC (non-small cell lung cancer) by ARMS (amplification refractory mutation system). Methods: Biopsy specimens from 181 cases of NSCLC were collected, in which 157 were small specimens (including specimens acquired through CT-guided percutaneous lung biopsy, endobronchial ultrasound-guided transbronchial needle aspiration, lymph node biopsy, bronchoscopic biopsy and aspiration of pleural effusion) and 24 were gross specimens. QIAGEN DNA extraction kit was used to extract DNAs from NSCLC specimens. Then AmoyDx EGFR Mutation Test Kit - a highly sensitive real-time PCR-based test was used to detect EGFR mutations. The detection rates of EGFR mutations in small specimens and gross specimens were compared by Chi-square test or Fisher's exact test. Results: The total EGFR mutation detection rate of all biopsy specimens was 39.8% (72/181). The detection rates of small specimens and gross specimens were 38.9% and 45.8%, respectively (P = 0.515). Furthermore, EGFR mutation frequencies were significantly higher in non-smokers (P = 0.033) and in patients with adenocarcinoma (P < 0.001). Conclusion: A relatively high detection rate of EGFR mutation in NSCLC small specimens can be obtained through ARMS. For patients with advanced NSCLC whose gross specimens cannot be easily obtained, the alternative small specimens can be used in clinical EGFR mutation detection. Copyright © 2012 by TUMOR.

Assessment of genetic mutations in the XRCC2 coding region by high resolution melting curve analysis and the risk of differentiated thyroid carcinoma in Iran

Homologous recombination (HR) is the major pathway for repairing double strand breaks (DSBs) in eukaryotes and XRCC2 is an essential component of the HR repair machinery. To evaluate the potential role of mutations in gene repair by HR in individuals susceptible to differentiated thyroid carcinoma (DTC) we used high resolution melting (HRM) analysis, a recently introduced method for detecting mutations, to examine the entire XRCC2 coding region in an Iranian population. HRM analysis was used to screen for mutations in three XRCC2 coding regions in 50 patients and 50 controls. There was no variation in the HRM curves obtained from the analysis of exons 1 and 2 in the case and control groups. In exon 3, an Arg188His polymorphism (rs3218536) was detected as a new melting curve group (OR: 1.46; 95 percentCI: 0.432-4.969; p = 0.38) compared with the normal melting curve. We also found a new Ser150Arg polymorphism in exon 3 of the control group. These findings suggest that genetic variations in the XRCC2 coding region have no potential effects on susceptibility to DTC. However, further studies with larger populations are required to confirm this conclusion.