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1.
Rev. peru. med. exp. salud publica ; 38(4): 577-586, oct.-dic. 2021. tab, graf
Article in Spanish | LILACS-Express | LILACS | ID: biblio-1365926

ABSTRACT

RESUMEN Objetivo. Determinar la estructura genética de las cepas drogorresistentes de Mycobacterium tuberculosis que circularon en todo el Perú durante los años 2011-2015 a través de haplotipos obtenidos de un ensayo con sondas en línea. Materiales y métodos. Se analizaron 6589 muestras que ingresaron al Instituto Nacional de Salud para el diagnóstico rutinario mediante el ensayo GenoType® MTBDRplus v2, durante el periodo de estudio. Se crearon haplotipos resistentes mediante la concatenación de 21 sitios polimórficos de los genes evaluados por el ensayo con sondas en línea, y se realizó el análisis de asociación con fenotipos obtenidos por el método de proporciones agar 7H10. Resultados. Las mutaciones de mayores frecuencias fueron: rpoB S531L (55,4%) y rpoB D516V (18,5%) para la resistencia a rifampicina, y katG S315T (59,5%) e inhA c-15t (25,7%) para la resistencia a isoniacida. Se obtuvieron 13 haplotipos representativos (87,8% de muestras analizadas) de los cuales seis correspondieron al genotipo multidrogorresistente, cuatro al genotipo monorresistente a isoniacida y tres al genotipo monorresistente a rifampicina. Dieciocho departamentos, y la provincia del Callao, presentaron una alta diversidad haplotípica; cuatro presentaron moderada diversidad y dos presentaron baja diversidad. Conclusiones. Existe una alta diversidad haplotípica en la mayoría de los departamentos, además de una concentración de las cepas de Mycobacterium tuberculosis drogorresistentes en las ciudades de Lima y Callao. Asimismo, las cepas de Mycobacterium tuberculosis con perfil drogorresistente que circulan en el Perú contienen principalmente los marcadores genéticos de mayor prevalencia a nivel mundial asociados con la resistencia frente a rifampicina e isoniacida.


ABSTRACT Objective. To determine the genetic structure of drug-resistant strains of Mycobacterium tuberculosis that circulated throughout Peru during the years 2011-2015, by using haplotypes obtained from a line probe assay. Materials and methods. A total of 6589 samples that were admitted to the Instituto Nacional de Salud for routine diagnosis using the GenoType® MTBDRplus v2 assay were analyzed during the study period. Resistant haplotypes were created by concatenating 21 polymorphic sites of the evaluated genes using the line probe assay; and the association analysis was carried out with phenotypes obtained by the 7H10 agar ratio method. Results. The most frequent mutations were: rpoB S531L (55.4%) and rpoB D516V (18.5%) for rifampicin resistance, and katG S315T (59.5%) and inhA c-15t (25.7%) for isoniazid resistance. We obtained 13 representative haplotypes (87.8% of analyzed samples), 6 corresponded to the multidrug-resistant genotype, 4 to the isoniazid mono-resistant genotype and 3 to the rifampicin mono-resistant genotype. Eighteen regions and the province of Callao showed high haplotype diversity; four showed moderate diversity and two showed low diversity. Conclusions. Most regions showed high haplotype diversity; in addition, most drug-resistant strains of Mycobacterium tuberculosis were concentrated in the cities of Lima and Callao. Likewise, drug-resistant Mycobacterium tuberculosis strains circulating in Peru mainly contain the genetic markers with the highest prevalence worldwide, which are associated with resistance to rifampicin and isoniazid.

2.
REVISA (Online) ; 10(2): 347-357, 2021.
Article in Portuguese | LILACS | ID: biblio-1224443

ABSTRACT

Objetivo: Analisar as implicações, impactos e o desenvolvimento de um indivíduo diagnosticado com TEA e portador da mutação de novo no gene DEAF1, a partir das várias perspectivas de intervenções realizadas. Método: Trata-se de um estudo descritivo com histórico dos tratamentos, resultados laboratoriais e genéticos mais recentes do paciente. Resultados: Sintomas notados aos 2 anos e diagnóstico específico aos 5. Aos 8 anos teve a primeira crise convulsiva tônico-clônica e o Eletroencefalograma alterado. Após obteve o diagnóstico molecular confirmado. Possuía epilepsia refratária de difícil controle, que houve piora com uma tentativa do uso de derivados canabinoides em conjunto com estimulação elétrica transcraniana. No momento, com os tratamentos, atendimentos multidisciplinares, dieta de exclusão de alérgenos e medicações de controle individual, diminuíram a intensidade das crises epiléticas e houve melhor controle do seu estado geral. Conclusão: Este estudo descreve como a mutação de novo no gene DEAF1 está relacionada com o TEA e com o comprometimento do desenvolvimento neurocognitivo. As terapias e métodos devem respeitar cada paciente na sua individualidade.


Objective: To analyze the implications, impacts and development of an individual diagnosed with ASD and carrying a de novo mutation in the DEAF1 gene, from the various perspectives of interventions performed. Method: This is a descriptive study, with the patient's history of treatments, and most recent laboratory and genetic results.Results: Symptoms were noticed at 2 years old and specific diagnosis at 5. At 8 years old he had his first tonic-clonic seizure and the electroencephalogram was altered. After, it was obtained the confirmed molecular diagnosis. He had refractory epilepsy that was difficult to control and aggravated with an attempt to use cannabinoid derivatives in conjunction with transcranial electrical stimulation. At the moment, treatments, multidisciplinary care, allergen exclusion diet and individual control medications, reduced the intensity of epileptic seizures and there was better control of his general condition. Conclusion: This study describes how the de novo mutation in the DEAF1 gene is related to ASD and neurocognitive development impairment. Therapies and methods must respect each patient in their individuality.


Objetivo: Analizar las implicaciones, impactos y desarrollo de un individuo diagnosticado de TEA y portador de una mutación de novo en el gen DEAF1, desde las distintas perspectivas de las intervenciones realizadas. Método: Este es un estudio descriptivo, con el historico de tratamientos del paciente y los resultados genéticos y de laboratorio más recientes. Resultados: Los síntomas se notaron a los 2 años y el diagnóstico específico a los 5. A los 8 años tuvo su primera crisis tónico-clónica y se alteró el electroencefalograma. Posteriormente se obtuvo el diagnóstico molecular confirmado. Tenía epilepsia refractaria que era difícil de controlar y se agravaba con un intento de utilizar derivados cannabinoides junto con estimulación eléctrica transcraneal. En el momento, los tratamientos, la atención multidisciplinar, la dieta de exclusión de alérgenos y los medicamentos de control individual, redujeron la intensidad de las crisis epilépticas y hubo un mejor control de su estado general. Conclusión: Este estudio describe cómo la mutación de novo en el gen DEAF1 se relaciona con el TEA y el deterioro del desarrollo neurocognitivo. Las terapias y los métodos deben respetar a cada paciente en su individualidad.


Subject(s)
Humans , Chromosomes, Human , Point Mutation , Autism Spectrum Disorder
3.
Article in Chinese | WPRIM | ID: wpr-908797

ABSTRACT

Objective:To evaluate the correlation between Card 9 gene polymorphism and acute pancreatitis(AP).Methods:70 AP patients and 70 healthy subjects from Shanghai Songjiang District Central Hospital from June 2019 to February 2020 were enrolled. TaqMan probe method was used to assay genotype distributions of the Card 9 polymorphisms rs10870077, rs4077515, rs10781499, rs141992399, rs139265120. Real-time quantitative PCR was used to determine the level of Card 9 mRNA, electrochemiluminescence immunoassay was applied to assay IL-6 and procalcitonin, and nephelometry was performed to measure the C-reactive protein(CRP).Results:Compared with the control group, the genotype and allele frequency of Card 9 gene rs10870077 C>G were significantly elevated in AP patients with statistically significant difference (31.4% vs 50.0%, P<0.05). There was no statistically significant difference on the allele frequency of Card 9 rs4077515AG and rs10781499AG, especially on rs141992399 C>G and rs139265120 A>G. C>Gpolymorphism in Card 9 rs10870077 resulted in an obvious increase of serum Card 9 mRNA expression in AP patients from 3.90±1.96 to 6.20±2.82, and the difference was statistically significant ( P<0.05), but there was no statistically significant difference on the Card 9 mRNA between AP patients with Card 9 rs4077515AG and rs10781499AG and the controls. IL level in AP patients with Card 9 rs10870077CG was greatly higher than that in those with Card 9 rs10870077 CC and GG [(614.7±1531.8 ng/L vs (372.5±1127.9)and (385.5±598.7)ng/L]. But compared with GG genotype, CRP level was obviously decreased [(34.84±50.64)mg/L vs (55.30±87.02)mg/L], and the procalcitonin was obviously increased [(1.98±4.70)μg/L vs (0.77±1.12)μg/L], and all the differences were statistically significant (all P value<0.05). Conclusions:Card 9 rs10870077 C>G mutation could upregulate the expression of Card 9 mRNA and increase IL-6 level, which may be a high risk for the occurrence of AP patients.

4.
Article in English | WPRIM | ID: wpr-879961

ABSTRACT

To establish a rabbit model of proprotein convertase subtilisin/kexin type9 () point mutation with CRISPR/Cas9 gene editing technique. According to the PubMed gene protein data, the PCSK9 protein functional regions of human and rabbit were analyzed by Blast. The 386S (Ser) amino acid functional region of human gene was homologous to the 485S of rabbit gene. Three small guide RNAs and one single-stranded donor oligonucleotide were designed according to the 485S base substitution position and sequence analysis of rabbit gene. The synthetic small guide RNAs, Cas9 mRNA and single-stranded donor oligonucleotide were co-injected into the cytoplasm of rabbit fertilized eggs and the embryos were transferred into the pregnant rabbits. PCR, TA cloning and off-target analysis were performed on the F0 rabbits to identify whether the PCSK9 mutation was successful. Fifteen F0 rabbits were obtained. The sequencing results showed that one of them was PCSK9 point mutation homozygote and two of them were PCSK9 point mutation heterozygotes, and the mutation could be stably inherited. The rabbit model of PCSK9 point mutation was successfully constructed by CRISPR/Cas9 technique, which provides an animal model for exploring the molecular mechanism of impaired PCSK9 function and developing reliable and effective diagnosis and treatment measures.


Subject(s)
Animals , CRISPR-Cas Systems/genetics , Mutation , Point Mutation , Proprotein Convertase 9/metabolism , Rabbits
5.
Journal of Clinical Hepatology ; (12): 809-812, 2021.
Article in Chinese | WPRIM | ID: wpr-875886

ABSTRACT

ObjectiveTo investigate the association of gene mutations in the pre-C, C, and basic core promoter (BCP) regions of hepatitis B virus (HBV) with traditional Chinese medicine (TCM) syndrome types in patients with chronic hepatitis B (CHB). MethodsA retrospective analysis was performed for the clinical data of CHB patients who were diagnosed and treated at the outpatient service and ward of Spleen, Stomach, and Hepatobiliary Department, The First Affiliated Hospital of Henan University of Chinese Medicine, from November 2014 to June 2018. Related clinical data were collected and recorded, including general information, HBV serological markers, HBV gene mutations, and information obtained by four TCM diagnostic methods. Syndrome differentiation and typing were performed for each patient with reference to the criteria for TCM syndrome differentiation of viral hepatitis, and the association of gene mutation in the pre-C, C, and BCP regions of HBV with TCM syndrome types was analyzed. The chi-square test was used for comparison of categorical data between groups, and the Kruskal-Wallis H test was used for comparison of continuous data between multiple or two groups. ResultsA total of 235 patients with CHB were enrolled, among whom 101 had internal retention of damp-heat, 88 had stagnation of liver Qi and spleen deficiency, 17 had blood stasis obstructing the collaterals, 19 had liver-kidney Yin deficiency, and 10 had spleen-kidney Yang deficiency. There were significant differences in sex, age, and course of disease between the patients with different TCM syndrome types (χ2=17.389, H=173.280, H=86.520, all P<0.01), and there was a significant difference in age between the CHB patients with gene mutations in the pre-C, C and BCP regions of HBV (H=30.150, P<0.001). There was a significant difference in the distribution of TCM syndrome types between the CHB patients with gene mutations in the pre-C, C and BCP regions of HBV (χ2=58.117, P<0.001), and internal retention of damp-heat and stagnation of liver Qi and spleen deficiency were major TCM syndrome types accounting for 80.43%. The patients with internal retention of damp-heat tended to have A1762T and G1764A mutations, and those with stagnation of liver Qi and spleen deficiency tended to have G1896A, A1762T, and G1764A mutations; G1764A mutation was often observed in the patients with blood stasis obstructing the collaterals or liver-kidney Yin deficiency, and I97L mutation was often observed in the patients with spleen-kidney Yang deficiency. ConclusionGene mutations in the pre-C, C, and BCP regions of HBV are associated with TCM syndrome types in CHB patients, and internal retention of damp-heat and stagnation of liver Qi and spleen deficiency are the most common TCM syndrome types. I97L mutation is often observed in patients with spleen-kidney Yang deficiency.

6.
J Genet ; 2020 Oct; 99: 1-7
Article | IMSEAR | ID: sea-215539

ABSTRACT

Rett syndrome (RTT) is an X-linked disorder caused by mutations in MECP2 in majority of cases. It is characterized by arrested development between 6 and 18 months of age, regression of acquired hand skills and speech, stereotypic hand movements, gait abnormalities and seizures. There are a very few studies in India which illustrates mutation spectrum in RTT. None of the studies have correlated seizures with the genotype. This study describes the phenotype and genotype spectrum in children with RTT syndrome and analyses the association of epilepsy with various clinical features and molecular findings. All children with RTT in our cohort had global developmental delay. Genetic diagnosis identified mutations of the MECP2 in all 25 children where RTT was suspected. We have identified point mutations in 20 patients, one insertion and four deletions by Sanger sequencing, namely c.1164_1207 (44 bp), c.1165_1207 (43 bp), c.1157_1197 (41 bp) del and c.1157_1188 (32 bp). Clinically, none of the patients with deletion had seizures. We identified one novel insertion variant c.337_338 (p.S113Ffs*9). All the deletions were located in the C-terminal region. Majority of the mutations (22/25) were identified in exon 4 which comprised of nonsense and missense types. Screening of hotspot mutations in exon 4 should be the first line evaluation in diagnosis of RTT. Molecular testing could help in specific management of seizures in RTT.

7.
Chinese Journal of Dermatology ; (12): 98-101, 2020.
Article in Chinese | WPRIM | ID: wpr-870229

ABSTRACT

A case of ichthyosis follicularis,alopecia and photophobia syndrome caused by a novel mutation c.1165C>T in the membrane-bound transcription factor protease site 2 (MBTPS2) gene was firstly reported.The proband presented with dry skin,congenital hairlessness,follicular keratotic papules,photophobia,epilepsy,and mental and motor retardation.Next-generation and Sanger sequencing analysis confirmed that the proband and his mother both had a c.1165C>T (p.pro389Ser) mutation in exon 9 of the MBTPS2 gene.According to the clinical manifestations of the patient and genetic characteristics of the MBTPS2 gene mutation,the patient was diagnosed with ichthyosis follicularis,alopecia and photophobia syndrome.

8.
Chinese Pharmacological Bulletin ; (12): 748-752, 2019.
Article in Chinese | WPRIM | ID: wpr-857219

ABSTRACT

The α1-adrenergic receptor (α1-AR) belongs to G-protein coupled receptors (GPCRs) and contains three subtypes, namely α1A, α1B and α1D. Studies have shown that structural changes in the α1-ARs receptor can lead to changes in their physiological functions. In the present study, we summarize the structural changes in four aspects of α1-ARs receptors, including carboxy-terminal truncation, dimerization, allosteric effects, and point mutations that lead to endocytosis, phosphorylation, desensitization, and affinity for agonists.

9.
Asian Journal of Andrology ; (6): 183-189, 2019.
Article in Chinese | WPRIM | ID: wpr-842578

ABSTRACT

Globozoospermia has been reported to be a rare but severe causation of male infertility, which results from the failure of acrosome biogenesis and sperm head shaping. Variants of dpy-19-like 2 (DPY19L2) are highly related to globozoospermia, but related investigations have been mainly performed in patients from Western countries. Here, we performed a screening of DPY19L2 variants in a cohort of Chinese globozoospermic patients and found that five of nine patients carried DPY19L2 deletions and the other four patients contained novel DPY19L2 point mutations, as revealed by whole-exome sequencing. Patient 3 (P3) contained a heterozygous variant (c.2126+5G>A), P6 contained a homozygous nonsense mutation (c.1720C>T, p.Arg574∗), P8 contained compound heterozygous variants (c.1182-1184delATC, p.Leu394-Ser395delinsPhe; c.368A>T, p.His123Arg), and P9 contained a heterozygous variant (c.1182-1184delATCTT, frameshift). We also reported intracytoplasmic sperm injection (ICSI) outcomes in the related patients, finding that ICSI followed by assisted oocyte activation (AOA) with calcium ionophore achieved high rates of live births. In summary, the infertility of these patients results from DPY19L2 dysfunction and can be treated by ICSI together with AOA.

10.
Article in Chinese | WPRIM | ID: wpr-821290

ABSTRACT

Objective@#To identify a α-globin gene mutation-IVS-Ⅱ-55 (T→A) and analyze hematological characteristics of IVS-Ⅱ-55 (T→G) carriers. @*Methods@#The peripheral blood samples were collected from the members of five family and three sporadic IVS-Ⅱ-55(T→G) carriers for the analysis of RBC parameters and hemoglobin electrophoresis. Gap-PCR, PCR-RDB (reverse dot blot) and DNA sequencing were carried out for the identification of gene deletion and mutation of α-globin and β-globin. @*Results@#The results of RBC parameters of five infant probands which presented with microcytic hypochromic anemia were below the normal reference interval. One of the adult carriers of IVS-Ⅱ-55 (T→G) heterozygote alone presented with microcytic hypochromic anemia, and the others showed normal RBC parameters. The hematological phenotype index (MCV, MCH and HbA 2 ) of one adult carrying a compound heterozygote for IVS-Ⅱ-55 (T→G) and βCD27-28M/N were 65.0 fL, 20.3 pg and 5.8% respectively. The hematological phenotype index (MCV, MCH, HbA 2 and HbF) of one adult carrying a compound heterozygote for IVS-Ⅱ-55 (T→G) and SEA-HPFH were 81.9 fL, 26.5 pg, 3.0% and 29.0% respectively. The HbA 2 levels of all carriers of IVS-Ⅱ-55 (T→G) heterozygote alone were in normal range. No abnormal hemoglobin band was detectable on hemoglobin electrophoresis for all the carries. @*Conclusion@#The carriers of IVS-Ⅱ-55(T→G) heterozygote alone were asymptomatic. The phenotype of compound heterozygote for β-thalassemia was similar to that of β-thalassemia alone.

11.
Tumor ; (12): 1-9, 2018.
Article in Chinese | WPRIM | ID: wpr-848429

ABSTRACT

Objective: To investigate the expression and mutation status of tensin family member 4 (TNS4) gene in various prostate cancer cell lines, and to explore its potential molecular mechanism. Methods: The expression of TNS4 gene in several prostate cancer cell lines was detected by RT-PCR. The full-length cDNA of TNS4 gene was amplified for the restriction endonuclease hydrolysis and DNA sequencing analysis. Meantime, the recombinant plasmids carrying wild- or mutant-type TNS4 gene were constructed and transfected into prostate cancer PC-3 cells, then the effects of TNS4 over-expression on the mRNA and protein expressions as well as the localization of signal transducer and activator of transcription 1 (STAT1) were detected by RT-PCR, Western blotting and immunofluorescence microscopy, respectively. Results: As compared with benign prostatic hyperplasia BPH1 cells, TNS4 gene was lowly expressed in prostate cancer cell lines DU145, LNCaP and 22Rv1 (all P 0.05). Additionally, the over-expression of mutant TNS4 significantly promoted nuclear translocation of STAT1 protein (P < 0.01). Conclusion: TNS4 gene is lowly expressed in prostate cancer cell lines, and there is a novel TNS4 mutant (S143P). TNS4 over-expression can up-regulate the expression of STAT1 protein, furthermore the mutant TNS4 can alter the location of STAT1 protein.

12.
Chinese Journal of Oncology ; (12): 35-39, 2018.
Article in Chinese | WPRIM | ID: wpr-809800

ABSTRACT

Objective@#To evaluate the consistency in detection of T790M mutation of epidermal growth factor receptor gene (EGFR) in plasma and tumor samples of patients with lung adenocarcinoma.@*Methods@#The tumor tissues or cytological specimens of 12 patients with operable lung adenocarcinoma(stage Ⅰ-ⅢA) and 100 patients with advanced stage ⅢB-Ⅳ lung adenocarcinoma were collected, among which 11 patients showed acquired resistance for gefitinib (11/100). In the same period, peripheral blood samples were collected from all patients and 50 healthy volunteers. Amplification refractory mutation system (ARMS) was used to detect EGFR mutations in tumor specimens. Next Generation Sequencing(NGS) based circulating single-molecule amplification and resequencing technology (cSMART)was performed to quantitatively detect the EGFR mutations in circulating tumor DNA (ctDNA) from plasma specimens.@*Results@#The sensitivity, specificity and concordance rate of EGFR T790M mutation between plasma and tissue specimens from 100 advanced stage patients were 50.0%, 72.9% and 72.0%, respectively. For L858R mutation and exon 19 deletion mutations, the above mentioned sensitivity, specificity and concordance rate were 91.7%, 100.0%, and 98.0%, as well as 79.2%, 100.0% and 95.0%, respectively. The L858R mutation and exon 19 deletion mutations were not detected in plasma of 50 healthy volunteers, whereasT790M mutation(1.0±0.0 copies) was found in 7 individuals(7/50, 14.0%). Similarly, in 12 resectable patients, 4 (4/12, 33.3%) T790M mutations were found in plasma (1.2±0.2 copies), but no L858R mutation and 19 exon deletion mutations. In comparison, 28.0% of patients with advanced lung adenocarcinoma (28/100)had detectable T790M mutation in plasma with copy numbers (34.0±22.7 copies). Furthermore, the copy numbers of T790M were 268.2±119.9 in plasma of 5 cases with acquired gefitinib-resistance.@*Conclusions@#In patients with advanced stages of lung adenocarcinoma, the detection of T790M mutation in plasma and tumor specimens is low. The T790M mutation also exists in the plasma of some healthy controls, suggesting that T790M mutation participates in EGFR signaling pathway and it might function in healthy population.

13.
Article in Chinese | WPRIM | ID: wpr-732724

ABSTRACT

Objective To study the correlation between coagulation factor Ⅴ Leiden mutation and ischemic stroke in Chinese young adults. Methods From July 2016 to January 2018, 80 young patients with ischemic stroke (18-45 years) admitted to the Department of Neurology, the First Affiliated Hospital of Shantou University Medical College and 80 controls were enrolled prospectively. The demographic data, vascular risk factors were documented. Sanger sequencing was used to detect factor Ⅴ Leiden mutation. Results The 1691 sites of factor Ⅴ genes in the case group and control group were all wild types. Multivariate logistic regression analysis suggested that hypertension ( odds ratio [ OR ] 4. 308, 95% confidence interval [CI] 3. 321-5. 067; P = 0. 001), hyperlipidemia (OR 2. 734, 95% CI 2. 214-3. 378; P = 0. 005), and smoking (OR 5. 293, 95% CI 3. 003-6. 180; P = 0. 010) were the independent risk factors for ischemic stroke in young adults, and high-density lipoprotein cholesterol (OR 0. 611, 95% CI 0. 457-0. 709; P = 0. 027) was its independent protective factor. Conclusion Factor Ⅴ Leiden mutation is not associated with ischemic stroke in Chinese young adults. Hypertension, dyslipidemia and smoking are still the main risk factors for ischemic stroke in young adults.

14.
Tianjin Medical Journal ; (12): 873-877, 2018.
Article in Chinese | WPRIM | ID: wpr-812971

ABSTRACT

@#Objective To summarize the clinical features of children with congenital Rett variant caused by mutation of FOXG1 and provide the reference for the diagnosis and treatment of the disease. Methods The clinical data of a patient diagnosed as congenital Rett syndrome variant type were summarized. The DNA samples of peripheral blood from the patient and her parents were extracted. The targeted high-throughput sequencing technology was used to detect the sequence of targeted genes, which were associated with the symptoms of the child. Genes were then verified by sanger sequencing. Chromosomal microarray analysis was performed to detect chromosome microdeletions and microduplications. Results The child carried the c.506dupG, p.G169Gfs* 286 heterozygous mutations on FOXG1 gene, which located in 14q12, and her parents were wild-type. After querying the HGMD, Clinvar and dbSNP databases, we found that it was not reported. This case was clearly diagnosed as congenital Rett syndrome variant type. We confirmed that the mutation locus was a new mutation. Conclusion For cases with congenital Rett variant manifestations, FOXG1 gene mutation examination is recommended, and preventive treatment of partially predictable dysfunction should be carried out.

15.
Einstein (Säo Paulo) ; 15(4): 492-495, Oct.-Dec. 2017. tab, graf
Article in English | LILACS | ID: biblio-891420

ABSTRACT

ABSTRACT Hereditary hyperferritinemia-cataract syndrome is an autosomal dominant genetic disorder associated with mutations in the 5'UTR region of the ferritin light chain gene. These mutations cause the ferritin levels to increase even in the absence of iron overload. Patients also develop bilateral cataract early due to accumulation of ferritin in the lens, and many are misdiagnosed as having hemochromatosis and thus not properly treated. The first cases were described in 1995 and several mutations have already been identified. However, this syndrome is still a poorly understood. We report two cases of unrelated Brazilian families with clinical suspicion of the syndrome, which were treated in our department. For the definitive diagnosis, the affected patients, their parents and siblings were submitted to Sanger sequencing of the 5'UTR region for detection of the ferritin light gene mutation. Single nucleotide polymorphism-like mutations were found in the affected patients, previously described. The test assisted in making the accurate diagnosis of the disease, and its description is important so that the test can be incorporated into clinical practice.


RESUMO A síndrome hereditária hiperferritinemia-catarata é uma doença genética autossômica dominante associada a mutações na região 5'UTR do gene da cadeia leve da ferritina. Estas mutações elevam os níveis de ferritina, mesmo na ausência de sobrecarga de ferro. Os pacientes também desenvolvem catarata bilateral precocemente, devido ao acúmulo de ferritina no cristalino, e muitos são erroneamente diagnosticados como portadores de hemocromatose, sendo tratados de maneira inadequada. Os primeiros casos foram descritos em 1995, e diversas mutações já foram identificadas. Entretanto, essa síndrome ainda é pouco conhecida. Relatamos dois casos de famílias brasileiras, não relacionadas, com suspeita clínica da síndrome, que foram atendidas em nosso serviço. Para o diagnóstico definitivo, os pacientes afetados, seus pais e irmãos foram submetidos à pesquisa de mutação do gene ferritina, por sequenciamento de Sanger da região 5'UTR. Foram encontradas mutações do tipo polimorfismo de nucleotídeo único nos pacientes afetados, já descritas anteriormente. O teste auxiliou no diagnóstico preciso da doença e é importante ser divulgado, para ser incorporado na prática clínica.


Subject(s)
Humans , Male , Child, Preschool , Child , Apoferritins/blood , Cataract/congenital , Iron Metabolism Disorders/congenital , Iron/blood , Syndrome , Cataract/genetics , Cataract/blood , Brazil , Iron Metabolism Disorders/genetics , Iron Metabolism Disorders/blood , Mutation/genetics
16.
Article in English | WPRIM | ID: wpr-819417

ABSTRACT

OBJECTIVE@#To investigate the antimicrobial susceptibility of 97 clinical Staphylococcus aureus (S. aureus) strains against 14 antimicrobials and corresponding resistance mechanisms.@*METHODS@#The antimicrobial susceptibility of the isolates was determined using a disk diffusion method and antimicrobial resistance genes were screened by polymerase chain reaction. Mutations responsible for ciprofloxacin and rifampicin resistance were investigated by polymerase chain reaction and DNA sequencing.@*RESULTS@#All isolates were found to be susceptible to vancomycin. Various rates of resistance to penicillin (83.5%), ampicillin (77.3%), erythromycin (63.9%), tetracycline (16.5%), amoxicillin/clavulanic acid (16.5%), ciprofloxacin (15.5%), trimethoprim/sulfamethoxazole (15.5%), oxacillin (13.4%), fusidic acid (12.4%), rifampin (6.2%), clindamycin (6.2%), gentamicin (6.2%) and mupirocin (5.2%) were determined. In addition, different combinations of resistance genes were identified among resistant isolates. Ciprofloxacin resistant isolates had mutations in codon 84 (Ser84Leu) and 106 (Gly106Asp) in the gyrA gene. Mutations in grlA were mostly related to Ser80Phe substitution. Leu466Ser mutation in the rpoB gene was detected in all rifampin resistant isolates. All methicillin resistant S. aureus isolates were SCCmec type V.@*CONCLUSIONS@#In conclusion, it was determined that the isolates were resistant to different classes of antimicrobials at varying rates and resistance was mediated by different genetic mechanisms. Therefore, continuous monitoring of resistance in S. aureus strains is necessary to control their resistance for clinically important antimicrobials.

17.
Article in Chinese | WPRIM | ID: wpr-512033

ABSTRACT

Objective To successfully get the full PCR alleles of the Insertion/Deletion marker rs10644346 in which a SNP-binding exists at the 3'end region of the primer.Methods Based on the AS-PCR,a common forward primer and two reverse primers with allele-specific oligonucleotides at the last second 3'end instead of the terminus were tentatively designed for typing 150 unrelated individuals and 10 father-mother--child trios from Htnan province in South-central China.Simultaneously,9 samples were typed with all the above three primers (the two primer sets which consist of the common forward primer and one of the reverse primers).Results PCR amplicons were well detected in the 150 unrelated individuals after being typed with the three primers,and the amplified fragments of parental and filial generations among 10 father-mother-child trios conformed to Mendel's principles.Allele missing was found in the two-primer group.Conclusion The primers designed by locating the specific nucleotide at the last second 3'end rather than terminal position were demonstrated also effective in getting specific alleles if perfect mismatch and PCR conditions are guaranteed,and the design strategy can provide an optional reference to rescue markers of SNP-binding primers for forensic practice.

18.
Article in Chinese | WPRIM | ID: wpr-510211

ABSTRACT

Objective To investigate the effect of G-quadruplex (G4) RNA structure of core of hepatitis C virus (HCV) on the specific immune response. Methods Circular dichroism (CD) was usedto detect the G4 spatial structure of the G4 oligonucleotide chain RNA (named as G4R) and its mutant of G4 (named as G4RM) by G base site-specific mutation.The HCV wild-type core gene G4(DNA) sequence was mutated as G4M-core by PCR site-directed mutagenesis without changing the amino acid codon.Then wild type and mutated core genes were constructed into the eukaryotic expression plasmid pcDNA3.1-Myc, and produced as pcDNA3.1-core-G4-WT (named as pG4) and pcDNA3.1-core-G4-M (named as pG4M), the expression of core protein was examined by Western blot. The mice were immunized with the pG4 and pG4M plasmids DNA respectively, and their humoral and cellular responses were examined. Results CD results showed that the structure of G4RM was changed compared to Wild type G4R, and the melting curve analysis showed the melting temperature of GR4M was lower than that of G4R, which indicates that G4RM structure is unstable. Western blot analysis showed that pG4M had much higher protein expression level compared to pG4(P<0.05). Analysis of animal immunization showed that pG4M induced increased levels of total IgG and IFN-γ compared to pG4(P<0.05). The IgG level of the pG4M group was 1.61 times higher than that of the pG4 group. By enzyme-linked immunospot(ELISpot)assay, we found that the release IFN-γ level of pG4M was 1.39 times higher than those of pG4. Flow cytometry showed that the intracellular IFN-γ production in the splenic CD4+ T cells was 1.79 times than those of pG4. Conclusion The G-quadruplex structure of HCV core can inhibit its protein translation. The mutation of G-quadruplex of core led to increased Th1-type immune responses. This is the first report demonstrate that HCV core G-quadruplex mutation can enhance its immunogenicity and could be used as a new strategy ofexploring HCV vaccine with enhanced immunogenicity.

19.
Article in Chinese | WPRIM | ID: wpr-506107

ABSTRACT

Objective To detect the sequence of SLC29A1 gene rs1288 single nucleotide polymorphism (SNP) in advanced pancreatic cancer patients,and to explore its correlation with gemcitabine toxicity and prognosis.Methods Peripheral blood samples were collected and DNA was extracted.The segment containing SLC29A1 gene SNP (rs1288) was amplified by PCR,and then DNA sequencing was conducted to identify SLC29A1 gene SNP (rs1288).According to the sequencing results,the patients were divided into SLC29A1 gene rs1288 T→A mutation type group and wild type group.Clinical data,toxicity of gemcitabine chemotherapy,progression free survival (PFS) and overall survival (OS) between two groups were compared.Results A total of 83 pancreatic cancer patients were enrolled.Sequencing results showed that SLC29 A1 gene 1288 T→A mutation type was present in 52 patients and wide type was observed in 31 patients,so mutation rate was 62.7%.All the patients in both two groups could tolerate the gemcitabine toxicity,and no chemotherapy related death occurred.There were no statistical differences on the gender,age,CA19-9,tumor site,size and TNM stage between the two groups.There were statistically higher iucidences of leukopenia and thrombocytopenia in the SLC29A1 gene rs1288 T →A mutation type group compared to the wild type group (55.8% vs 32.3%,P<0.05;40.4% vs 19.4%,both P<0.05).Median OS and PFS in mutation type group were shorter than those in wide type group (11 months vs 14 months,P < 0.05;9 months vs 12 months,P < 0.05).Conclusions Advanced pancreatic cancer patients with the SLC29A1 gene rs1288 T→A mutation type had a higher incidence of adverse reaction in gemcitabine chemotherapy and a worse therapeutic effect,and thus detecting the mutation of SLC29A1 gene rs1288 point mutation may serve as a marker for evaluating the toxicity and prognosis of gemcitabine chemotherapy in patients with pancreatic cancer.

20.
Article in Chinese | WPRIM | ID: wpr-333423

ABSTRACT

Genetic mutations are important molecular biomarkers for cancer diagnosis and surveillance.Therefore,the development of methods for mutation detection characterized with straightforward,highly specific and sensitive to low-level mutations within various sequence contexts is extremely needed.Although some of the currently available methods have shown very encouraging results,their discrimination efficiency is still very low.Herein,we demonstrate a fluorescent probe coupled with blocker and property of melting temperature discrimination,which is able to identify the presence of known or unknown single-base variations at abundances down to 0.1% within 20 min.The discrimination factors between the perfect-match target and single-base mismatched target are determined to be 10.15-38.48.The method is sequence independent,which assures a wide range of application.The new method would be an ideal choice for high-throughput in vitro diagnosis and precise clinical treatment.

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