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1.
Exp Cell Res ; 422(1): 113439, 2023 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-36464122

RESUMO

Colorectal cancer (CRC) is a common malignant tumor of digestive system. Its incidence rate and mortality rate ranks the third among all the malignant tumors. The objective of this study was to explore the role of ß-catenin in the CRC progression. The CRC tissues were collected to analyze the ß-catenin levels. The CRC cells (SW620 and RRKO) were treated with hypoxia to simulate the hypoxic microenvironment of tumor in vitro. The ß-catenin levels in the CRC cells were assessed with RT-qPCR, Western blot and Immunofluorescence. The cell biological behaviors were determined with CCK-8, flow cytometry and sphere formation assays. Besides, the glucose uptake, lactate production, ECAR and OCR was detected by seahorse. For the ß-catenin lactylation determination, the IP and Western blot assay was performed. Then the protein stability of ß-catenin was measured after cycloheximide treatment. The results showed that ß-catenin was highly expressed in the CRC tissues and cells. Hypoxia treatment dramatically increased the protein levels and lactylation of ß-catenin in the CRC cells. In addition, ß-catenin knockdown dramatically inhibited the cell growth and stemness of the CRC cells. Besides, activation of Wnt signaling pathway neutralized the role of sh-ß-catenin in the hypoxia treated CRC cells. In conclusion, this study confirmed that hypoxia induced the glycolysis promoted the ß-catenin lactylation, which further enhanced the protein stability and expression of ß-catenin, thus aggravating the malignant behaviors of CRC cells.


Assuntos
Neoplasias Colorretais , beta Catenina , Humanos , beta Catenina/genética , beta Catenina/metabolismo , Via de Sinalização Wnt , Neoplasias Colorretais/patologia , Linhagem Celular Tumoral , Proliferação de Células , Hipóxia , Regulação Neoplásica da Expressão Gênica , Movimento Celular , Microambiente Tumoral
2.
Environ Res ; 252(Pt 1): 118860, 2024 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-38582422

RESUMO

The application of antimony sulfide sensors, characterized by their exceptional stability and selectivity, is of emerging interest in detection research, and the integration of graphitized carbon materials is expected to further enhance their electrochemical performance. This study represents a pioneering effort in the synthesis of carbon-doped antimony sulfide materials through the pyrolysis of the mixture of microorganisms and their synthetic antimony sulfide. The prepared materials are subsequently applied to electrochemical sensors for monitoring the highly toxic compounds catechol (CC) and hydroquinone (HQ) in the environment. Via cyclic voltammetry (CV) and impedance testing, we concluded that the pyrolytic product at 700 °C (Sb-700) demonstrated the best electrochemical properties. Differential pulse voltammetry (DPV) revealed impressive separation when utilizing Sb-700/GCE for simultaneous detection of CC and HQ, exhibiting good linearity within the concentration range of 0.1-140 µM. The achieved sensitivities of 24.62 µA µM-1 cm-2 and 22.10 µA µM-1 cm-2 surpassed those of most CC and HQ electrochemical sensors. Meanwhile, the detection limits for CC and HQ were as low as 0.18 µM and 0.16 µM (S/N = 3), respectively. Additional tests confirmed the good selectivity, reproducibility, and long-term stability of Sb-700/GCE, which was effective in detecting CC and HQ in tap water and river water, with recovery rates of 100.7%-104.5% and 96.5%-101.4%, respectively. It provides a method that combines green microbial synthesis and simple pyrolysis for the preparation of electrode materials in CC and HQ electrochemical sensors, and also offers a new perspective for the application of microbial synthesized materials.


Assuntos
Antimônio , Catecóis , Técnicas Eletroquímicas , Hidroquinonas , Pirólise , Hidroquinonas/química , Hidroquinonas/análise , Catecóis/análise , Catecóis/química , Antimônio/química , Antimônio/análise , Técnicas Eletroquímicas/métodos , Técnicas Eletroquímicas/instrumentação , Sulfetos/química
3.
Biotechnol Bioeng ; 120(12): 3622-3637, 2023 12.
Artigo em Inglês | MEDLINE | ID: mdl-37691180

RESUMO

S-adenosyl- l-methionine (SAM) is a high-value compound widely used in the treatment of various diseases. SAM can be produced through fermentation, but further enhancing the microbial production of SAM requires novel high-throughput screening methods for rapid detection and screening of mutant libraries. In this work, an SAM-OFF riboswitch capable of responding to the SAM concentration was obtained and a high-throughput platform for screening SAM overproducers was established. SAM synthase was engineered by semirational design and directed evolution, which resulted in the SAM2S203F,W164R,T251S,Y285F,S365R mutant with almost twice higher catalytic activity than the parental enzyme. The best mutant was then introduced into Saccharomyces cerevisiae BY4741, and the resulting strain BSM8 produced a sevenfold higher SAM titer in shake-flask fermentation, reaching 1.25 g L-1 . This work provides a reference for designing biosensors to dynamically detect metabolite concentrations for high-throughput screening and the construction of effective microbial cell factories.


Assuntos
Riboswitch , S-Adenosilmetionina , S-Adenosilmetionina/metabolismo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Ensaios de Triagem em Larga Escala , Riboswitch/genética , Fermentação
4.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 40(4): 423-428, 2023 Apr 10.
Artigo em Zh | MEDLINE | ID: mdl-36972936

RESUMO

OBJECTIVE: To explore the genetic basis for 7 families with gonadal mosaicism for Duchenne muscular dystrophy (DMD). METHODS: For the 7 families presented at the CITIC Xiangya Reproductive and Genetic Hospital from September 2014 to March 2022, clinical data were collected. Preimplantation genetic testing for monogenic disorders (PGT-M) was carried out for the mother of the proband from family 6. Peripheral venous blood samples of the probands, their mothers and other patients from the families, amniotic fluid samples from families 1 ~ 4 and biopsied cells of embryos cultured in vitro from family 6 were collected for the extraction of genomic DNA. Multiplex ligation-dependent probe amplification (MLPA) was carried out for the DMD gene, and short tandem repeat (STR)/single nucleotide polymorphism (SNP)-based haplotypes were constructed for the probands, other patients, fetuses and embryos. RESULTS: The results of MLPA showed that the probands and the fetuses/probands' brothers in families 1 ~ 4, 5, 7 had carried the same DMD gene variants, whilst the probands' mothers were all normal. The proband in family 6 carried the same DMD gene variant with only 1 embryo (9 in total) cultured in vitro, and the DMD gene of the proband's mother and the fetus obtained through the PGT-M were normal. STR-based haplotype analysis showed that the probands and the fetuses/probands' brothers in families 1 ~ 3 and 5 have inherited the same maternal X chromosome. SNP-based haplotype analysis showed that the proband from family 6 has inherited the same maternal X chromosome with only 1 embryo (9 in total) cultured in vitro. The fetuses in families 1 and 6 (via PGT-M) were both confirmed to be healthy by follow up, whilst the mothers from families 2 and 3 had chosen induced labor. CONCLUSION: Haplotype analysis based on STR/SNP is an effective method for judging gonad mosaicism. Gonad mosaicisms should be suspected for women who have given births to children with DMD gene variants but with a normal peripheral blood genotype. Prenatal diagnosis and reproductive intervention may be adapted to reduce the births of further affected children in such families.


Assuntos
Distrofia Muscular de Duchenne , Masculino , Gravidez , Criança , Humanos , Feminino , Distrofia Muscular de Duchenne/genética , Distrofia Muscular de Duchenne/diagnóstico , Distrofina/genética , Mosaicismo , Éxons , Diagnóstico Pré-Natal/métodos , Nucleotídeos
5.
Yi Chuan ; 45(1): 88-95, 2023 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-36927641

RESUMO

Duchenne/Becker muscular dystrophy (DMD/BMD) is one of the most common progressive muscular dystrophy diseases with X-linked recessive inheritance. It is mainly caused by the deletion, duplication and point mutation of DMD gene. In rare cases, it is also caused by the destruction of DMD gene by chromosomal structural rearrangement. Here, we report a case of Duchenne/Becker Muscular dystrophy (DMD/BMD) with typical symptoms but unknown genetic defects after MLPA and next generation sequencing tests in other hospitals. Interestingly, we find a pericentric inversion of X chromosome (Chr.X: g. [31939463-31939465del; 31939466-131765063 inv; 131765064-131765067del]) in this patient. We then use the karyotyping, FISH, long-read sequencing and Sanger sequencing technologies to characterize the chromosome rearrangement. We find that this chromosomal aberration disrupt both the DMD gene and the HS6ST2 gene. The patient present with typical DMD symptoms such as muscle weakness, but no obvious symptoms of Paganini-Miozzo syndrome. Our results suggest that the destruction of DMD gene by structural rearrangement is also one of the important causes of DMD. Therefore, we suggest to provide further genetic testing for those DMD patients with unknown genetic defects through routine genetic testing. Cost-effective karyotyping and FISH should be considered firstly to identify chromosome rearrangements. Long-read sequencing followed by Sanger sequencing could be useful to locate the precise breakpoints. The genetic diagnosis of this case made it possible for reproductive intervention in the patient's family.


Assuntos
Distrofia Muscular de Duchenne , Humanos , Distrofia Muscular de Duchenne/genética , Distrofia Muscular de Duchenne/diagnóstico , Distrofina/genética , Testes Genéticos , Rearranjo Gênico/genética , Cromossomo X , Sulfotransferases/genética
6.
Mol Biol Evol ; 38(2): 486-501, 2021 01 23.
Artigo em Inglês | MEDLINE | ID: mdl-32946576

RESUMO

Bumblebees are a diverse group of globally important pollinators in natural ecosystems and for agricultural food production. With both eusocial and solitary life-cycle phases, and some social parasite species, they are especially interesting models to understand social evolution, behavior, and ecology. Reports of many species in decline point to pathogen transmission, habitat loss, pesticide usage, and global climate change, as interconnected causes. These threats to bumblebee diversity make our reliance on a handful of well-studied species for agricultural pollination particularly precarious. To broadly sample bumblebee genomic and phenotypic diversity, we de novo sequenced and assembled the genomes of 17 species, representing all 15 subgenera, producing the first genus-wide quantification of genetic and genomic variation potentially underlying key ecological and behavioral traits. The species phylogeny resolves subgenera relationships, whereas incomplete lineage sorting likely drives high levels of gene tree discordance. Five chromosome-level assemblies show a stable 18-chromosome karyotype, with major rearrangements creating 25 chromosomes in social parasites. Differential transposable element activity drives changes in genome sizes, with putative domestications of repetitive sequences influencing gene coding and regulatory potential. Dynamically evolving gene families and signatures of positive selection point to genus-wide variation in processes linked to foraging, diet and metabolism, immunity and detoxification, as well as adaptations for life at high altitudes. Our study reveals how bumblebee genes and genomes have evolved across the Bombus phylogeny and identifies variations potentially linked to key ecological and behavioral traits of these important pollinators.


Assuntos
Adaptação Biológica/genética , Abelhas/genética , Evolução Biológica , Genoma de Inseto , Animais , Uso do Códon , Elementos de DNA Transponíveis , Dieta , Comportamento Alimentar , Componentes do Gene , Tamanho do Genoma , Seleção Genética
7.
Int J Mol Sci ; 23(22)2022 Nov 18.
Artigo em Inglês | MEDLINE | ID: mdl-36430757

RESUMO

As important pollinators, bees play a critical role in maintaining the balance of the ecosystem and improving the yield and quality of crops. However, in recent years, the bee population has significantly declined due to various pathogens and environmental stressors including viruses, bacteria, parasites, and increased pesticide application. The above threats trigger or suppress the innate immunity of bees, their only immune defense system, which is essential to maintaining individual health and that of the colony. In addition, bees can be divided into solitary and eusocial bees based on their life traits, and eusocial bees possess special social immunities, such as grooming behavior, which cooperate with innate immunity to maintain the health of the colony. The omics approach gives us an opportunity to recognize the distinctive innate immunity of bees. In this regard, we summarize innate bee immunity from a genomic and transcriptomic perspective. The genetic characteristics of innate immunity were revealed by the multiple genomes of bees with different kinds of sociality, including honeybees, bumblebees, wasps, leaf-cutter bees, and so on. Further substantial transcriptomic data of different tissues from diverse bees directly present the activation or suppression of immune genes under the infestation of pathogens or toxicity of pesticides.


Assuntos
Praguicidas , Transcriptoma , Abelhas/genética , Animais , Ecossistema , Genômica , Imunidade Inata/genética , Comportamento Social
8.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 39(9): 925-931, 2022 Sep 10.
Artigo em Zh | MEDLINE | ID: mdl-36082559

RESUMO

OBJECTIVE: To summarize the genetic characteristics of 671 Chinese pedigrees affected with Duchenne/Becker muscular dystrophy (DMD/BMD). METHODS: Clinical data of the pedigrees were collected. Multiplex PCR, multiple ligation dependent probe amplification (MLPA), next generation sequencing (NGS), Sanger sequencing and long read sequencing were used to detect the variant of DMD gene in the probands and their mothers, and prenatal diagnosis was provided for high risk pregnant women. RESULTS: Among 178 pedigrees analyzed by multiplex PCR, 44 variants of the DMD gene were detected, with the genetic diagnosis attained in 110 pedigrees. Among 493 pedigrees analyzed by MLPA in combination with NGS or Sanger sequencing, 294 pathogenic/possible pathogenic variants were identified, among which 45 were unreported previously, and the genetic diagnosis attained in 484 pedigrees. Structural variants of the DMD gene were identified in two pedigrees by long-read sequencing. Among 444 probands, 341 have inherited the DMD gene variant from their mothers (76.8%). Among 390 women with a high-risk, 339 have opted to have natural pregnancy and 51 chose preimplantation genetic testing for monogenetic disease (PGT-M). The detection rate of neonatal patients and carriers following natural pregnancy was significantly higher than that for PGT-M. CONCLUSION: Combined application of MLPA, NGS, Sanger sequencing and long-read sequencing is an effective strategy to detect DMD/BMD. PGT-M can effectively reduce the risk of fetuses. Above finding has expanded the spectrum of DMD gene variants and provided a basis for reproductive intervention for pregnancies with a high risk for DMD/BMD.


Assuntos
Distrofia Muscular de Duchenne , China , Distrofina/genética , Éxons , Feminino , Testes Genéticos , Humanos , Recém-Nascido , Reação em Cadeia da Polimerase Multiplex , Distrofia Muscular de Duchenne/diagnóstico , Distrofia Muscular de Duchenne/genética , Mutação , Linhagem , Gravidez , Diagnóstico Pré-Natal
9.
J Immunol ; 202(1): 249-259, 2019 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-30510067

RESUMO

The peptidoglycan recognition protein SAs (PGRP-SAs) from Bombus ignitus (Bi-PGRP-SA), Apis mellifera (Am-PGRP-SA), and Megachile rotundata PGRP-SA (Mr-PGRP-SA) exhibit an intrinsic ability to preferentially bind to Dap-type peptidoglycan (PGN) from Bacillus subtilis rather than Lys-type PGN from Micrococcus luteus This ability is more analogous to the binding exhibited by PGRP-LCx and PGRP-SD than to that exhibited by PGRP-SA in Drosophila Moreover, Bi-PGRP-SA and Am-PGRP-SA share greater sequence identity with Drosophila PGRP-LCx than with PGRP-SD and retain several conserved contact residues, including His37/His38, His60/His61, Trp66/Trp67, Ala150/Ala151, and Thr151/Thr152 However, the corresponding contact residue Arg85 is not a major anchor residue in bees (e.g., bumblebees, honeybees, and leaf-cutting bees), and an in silico analysis indicated that the residues Thr151/Thr152 and Ser153/Ser154 of Bi-PGRP-SA and Am-PGRP-SA are deduced to be anchor residues. In addition, the nonconserved residues Asp67 in Bi-PGRP-SA and Mr-PGRP-SA and His68 in Am-PGRP-SA are deduced to be involved in the binding to Dap-type PGNs in bumblebees, honeybees, and leaf-cutting bees. We conclude that the structures and specificities of PGRP-SAs in bees are more analogous to those of PGRP-LCx than to those of Drosophila PGRP-SA. This phenomenon might be explained by the fact that the evolutionary clade of Hymenoptera is more ancient than that of Diptera.


Assuntos
Bacillus subtilis/fisiologia , Abelhas , Proteínas de Transporte/genética , Proteínas de Insetos/metabolismo , Micrococcus luteus/fisiologia , Peptidoglicano/metabolismo , Domínios e Motivos de Interação entre Proteínas/genética , Animais , Sítios de Ligação/genética , Evolução Biológica , Proteínas de Transporte/química , Proteínas de Transporte/metabolismo , Cristalografia por Raios X , Drosophila , Proteínas de Insetos/química , Proteínas de Insetos/genética , Modelos Químicos , Peptidoglicano/química , Filogenia , Ligação Proteica , Homologia de Sequência , Relação Estrutura-Atividade
10.
Int J Clin Pract ; 75(2): e13673, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32791569

RESUMO

OBJECTIVE: We aimed to evaluate oblique-axis in-plane (OA-IP) techniques for real-time ultrasound-guided internal jugular vein (IJV) cannulation. METHODS: We retrospectively analysed 1065 patients who underwent ultrasound (US)-guided IJV cannulation. We recorded demographic characteristics of patients, success rate, access time, cannulation time, number of attempts and the incidence of acute complications. RESULTS: The overall success rate of the procedure was 100% (n = 1605). In total, 1594 cases (99.3%) were successful at the first attempt, and 11 (0.7%) were successful at the second attempt; no patient required three or more attempts. The mean access time was 18.7 ± 19.3 seconds. The mean cannulation time was 349.0 ± 103.8 seconds. There were 54 (3.4%) acute complications out of the total 1605 cannulations: 23 cases of puncture site bleeding (1.4%), 20 cases allergic to dressing (1.3%), 10 cases of local cervical hematomas (0.6%), and one catheter misplacement (0.1%). There were no major complications 12 hours following the procedure. CONCLUSIONS: The results of our study suggest that OA-IP techniques can improve ultrasound-guided IJV cannulation with a high success rate and safety in clinical practice. Clinicians should consider adopting these methods.


Assuntos
Cateterismo Venoso Central , Veias Jugulares , Cateterismo Venoso Central/efeitos adversos , Hospitais , Humanos , Veias Jugulares/diagnóstico por imagem , Estudos Prospectivos , Estudos Retrospectivos , Ultrassonografia de Intervenção , Universidades
11.
Proc Natl Acad Sci U S A ; 115(24): 6249-6254, 2018 06 12.
Artigo em Inglês | MEDLINE | ID: mdl-29760103

RESUMO

Our understanding of phylogenetic relationships among bony fishes has been transformed by analysis of a small number of genes, but uncertainty remains around critical nodes. Genome-scale inferences so far have sampled a limited number of taxa and genes. Here we leveraged 144 genomes and 159 transcriptomes to investigate fish evolution with an unparalleled scale of data: >0.5 Mb from 1,105 orthologous exon sequences from 303 species, representing 66 out of 72 ray-finned fish orders. We apply phylogenetic tests designed to trace the effect of whole-genome duplication events on gene trees and find paralogy-free loci using a bioinformatics approach. Genome-wide data support the structure of the fish phylogeny, and hypothesis-testing procedures appropriate for phylogenomic datasets using explicit gene genealogy interrogation settle some long-standing uncertainties, such as the branching order at the base of the teleosts and among early euteleosts, and the sister lineage to the acanthomorph and percomorph radiations. Comprehensive fossil calibrations date the origin of all major fish lineages before the end of the Cretaceous.


Assuntos
Peixes/genética , Genoma/genética , Transcriptoma/genética , Animais , Evolução Molecular , Éxons/genética , Fósseis , Duplicação Gênica/genética , Genômica/métodos , Modelos Genéticos , Filogenia
12.
Korean J Physiol Pharmacol ; 25(4): 261-272, 2021 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-34187945

RESUMO

Doxorubicin (Dox) is widely used to the treatment of cancer, however, it could cause damage to gastric mucosa. To investigate the protective effects and related mechanisms of coenzyme Q10 (CoQ10) and vitamin C (VC) on Dox-induced gastric mucosal injury, we presented the survey of the 4 groups of the rats with different conditions. The results showed Dox treatment significantly induced GES-1 apoptosis, but preconditioning in GES-1 cells with VC or CoQ10 significantly inhibited the Dox-induced decrease and other harm effects, including the expression and of IκKß, IκBα, NF-κB/p65 and tumor necrosis factor (TNF-α) in GES-1 cells. Moreover, high-throughput sequencing results showed Dox treatment increased the number of harmful gut microbes, and CoQ10 and VC treatment inhibited this effect. CoQ10 and VC treatment inhibits Dox-induced gastric mucosal injury by inhibiting the activation of the IkKB/IκBα/NF-κB/p65/TNF-α pathway, promoting anti-inflammatory effects of gastric tissue and regulating the composition of the intestinal flora.

13.
J Exp Bot ; 71(1): 435-449, 2020 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-31616940

RESUMO

Soluble carbohydrates not only directly affect plant growth and development but also act as signal molecules in processes that enhance tolerance to cold stress. Raffinose family oligosaccharides (RFOs) are an example and play an important role in abiotic stress tolerance. This study aimed to determine whether galactinol, a key limiting factor in RFO biosynthesis, functions as a signal molecule in triggering cold tolerance. Exposure to low temperatures induces the expression of galactinol synthase (AnGolS1) in Ammopiptanthus nanus, a desert plant that survives temperatures between -30 °C to 47 °C. AnGolS1 has a greater catalytic activity than tomato galactinol synthase (SlGolS2). Moreover, SlGolS2 is expressed only at low levels. Expression of AnGolS1 in tomato enhanced cold tolerance and led to changes in the sugar composition of the seeds and seedlings. AnGolS1 transgenic tomato lines exhibited an enhanced capacity for ethylene (ET) signaling. The application of galactinol abolished the repression of the ET signaling pathway by 1-methylcyclopropene during seed germination. In addition, the expression of ERF transcription factors was increased. Galactinol may therefore act as a signal molecule affecting the ET pathway.


Assuntos
Resposta ao Choque Frio/genética , Fabaceae/genética , Galactosiltransferases/genética , Proteínas de Plantas/genética , Solanum lycopersicum/fisiologia , Dissacarídeos/metabolismo , Fabaceae/enzimologia , Galactosiltransferases/metabolismo , Solanum lycopersicum/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo
14.
Genomics ; 111(3): 222-230, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30465915

RESUMO

Southern catfish (Silurus meridionalis) is an economically important species widely cultured in China. It is well known for its fast growth, strong resistance to diseases and euryphage. However, little is known about the mechanisms for its powerful immune systems. Our Fish-T1K project has finished its first phase of 200 fish transcriptomes, with sequencing of gills in most examined fishes. In this study, we performed transcriptome sequencing of the gill and the maxillary barbel of Southern catfish, with the latter as a control. High expression of immune-related transcripts were observed in these two tissues. We observed that genes in the T cell receptor signaling pathway had higher transcription values in the gill than in the barbel. In addition, eight new rhamnose-binding lectins (RBLs) were identified and their carbohydrate recognition domains (CRDs) were classified according to the eight conserved cysteine residues and two conserved motifs (-YGR- and -DPC-). This is the first transcriptome report by high-throughput sequencing of the Southern catfish. Our genomic data and discovery of novel RBLs in this project should be able to promote better understandings of the roles of gills in immune responses and disease prevention for further aquaculture.


Assuntos
Proteínas de Peixes/genética , Peixes/genética , Lectinas/genética , Transcriptoma , Animais , Sequência Conservada , Proteínas de Peixes/química , Proteínas de Peixes/imunologia , Peixes/imunologia , Brânquias/imunologia , Brânquias/metabolismo , Lectinas/química , Lectinas/imunologia , Receptores de Antígenos de Linfócitos T/imunologia
15.
J Hum Genet ; 64(3): 207-214, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30617278

RESUMO

Autosomal recessive polycystic kidney disease (ARPKD), is a rare hepatorenal fibrocystic disorder primarily associated with progressive growth of multiple cysts in the kidneys causing progressive loss of renal function. The disease is linked to mutations in the PKHD1 gene. In this study, we describe the gene diagnosis and prenatal diagnosis for a consanguineous family with two fetuses diagnosed with polycystic kidney disease by fetal sonography during the pregnancy. Sequence analysis of cDNA synthesized from the PKHD1 mRNA of the second induced fetus identified a 111-nucleotide insert at the junction of exon 56 and 57 that originated from intervening sequence (IVS) 56. Further genomic sequencing of IVS 56 of the PKHD1 gene identified a rare homozygous deep intronic mutation (c.8798-459 C > A), which was inherited from the parents and not detectable in 100 unrelated control subjects. Moreover, we explored the pathogenicity of this deep intronic mutation by conducting a minigene splicing assay experiment, which demonstrated that the mutation causes a pseudoexon insertion, which results in a frameshift followed by a premature termination codon in exon 57. Eventually, the parents had a healthy baby by undergoing prenatal genetic diagnosis based on the targeted detection of the intron mutation. The newly identified deep intronic mutation is associated with a rare mechanism of abnormal splicing that expands the spectrum of known PKHD1 gene mutations. It can be used in evidence-based genetic and reproductive counseling for families with ARPKD.


Assuntos
Éxons , Íntrons , Mutação , Rim Policístico Autossômico Recessivo/genética , Receptores de Superfície Celular/genética , Feminino , Testes Genéticos , Genótipo , Humanos , Recém-Nascido , Masculino , Fenótipo , Rim Policístico Autossômico Recessivo/diagnóstico , Gravidez , Resultado da Gravidez , Diagnóstico Pré-Natal
16.
Nanotechnology ; 30(49): 495501, 2019 Dec 06.
Artigo em Inglês | MEDLINE | ID: mdl-31443101

RESUMO

With the capability of inducing small particle sizes of supported metal in graphite oxide (GO), the γ-ray irradiation method applied for preparing graphite oxide-gold (GO-Au) nanocomposites as electrochemical immunosensors has attracted specific attention recently. To study the accurate factors influencing the precise morphology and final performance of the prepared composites in the γ-irradiation system, we proposed a facile method to investigate the evolution of the GO structure, size and dispersion of Au nanoparticles (AuNPs) produced with the addition of isopropyl alcohol to the system. The GO-Au nanocomposites were characterized by Fourier transform infrared spectroscopy, x-ray diffraction spectra, Raman spectra, x-ray photoelectron spectroscopy and high resolution transmission electron microscopy. These nanocomposites with sandwich morphology exhibited an excellent immunosensor performance with a low detection limit of 15.8 pg ml-1 (S/N = 3) and a wide linear range from 1 to 40 ng ml-1 for detecting carcinoembryonic antigens. The enhanced biosensing performance is attributed to the synergistic effect of γ-irradiation and the precise structure of GO, which endows the smaller size and more uniform distribution of AuNPs on the GO as well as the good signal amplification capability. Furthermore, adopting the γ-irradiation method and use of GO as a precursor is propitious for application in large-scale production because of its high-efficiency and high-yielding characteristics.


Assuntos
Técnicas Biossensoriais , Antígeno Carcinoembrionário/sangue , Ouro/química , Imunoensaio , Nanopartículas Metálicas/química , Nanocompostos/química , 2-Propanol/química , Anticorpos Imobilizados/química , Técnicas Eletroquímicas , Raios gama , Grafite/química , Humanos , Limite de Detecção , Nanopartículas Metálicas/efeitos da radiação , Nanopartículas Metálicas/ultraestrutura , Nanocompostos/efeitos da radiação , Nanocompostos/ultraestrutura , Óxidos/química
17.
BMC Med Genet ; 19(1): 186, 2018 10 17.
Artigo em Inglês | MEDLINE | ID: mdl-30333007

RESUMO

BACKGROUND: Autosomal dominant polycystic kidney disease (ADPKD), the commonest inherited kidney disease, is generally caused by heterozygous mutations in PKD1, PKD2, or GANAB (PKD3). METHODS: We performed mutational analyses of PKD genes to identify causative mutations. A set of 90 unrelated families with ADPKD were subjected to mutational analyses of PKD genes. Genes were analysed using long-range PCR (LR-PCR), direct PCR sequencing, followed by multiplex ligation-dependent probe amplification (MLPA) or screening of GANAB for some patients. Semen quality was assessed for 46 male patients, and the correlation between mutations and male infertility was analysed. RESULTS: A total of 76 mutations, including 38 novel mutations, were identified in 77 families, comprising 72 mutations in PKD1 and 4 in PKD2, with a positive detection rate of 85.6%. No pathogenic mutations of GANAB were detected. Thirty-seven patients had low semen quality and were likely to be infertile. No association was detected between PKD1 mutation type and semen quality. However, male patients carrying a pathogenic mutation in the Ig-like repeat domain of PKD1 had a high risk of infertility. CONCLUSION: Our study identified a group of novel mutations in PKD genes, which enrich the PKD mutation spectrum and might help clinicians to make precise diagnoses, thereby allowing better family planning and genetic counselling. Men with ADPKD accompanied by infertility should consider intracytoplasmic sperm injection combined with preimplantation genetic diagnosis to achieve paternity and obtain healthy progeny.


Assuntos
Predisposição Genética para Doença , Infertilidade Masculina/genética , Mutação , Rim Policístico Autossômico Dominante/genética , Canais de Cátion TRPP/genética , Adulto , Povo Asiático , Análise Mutacional de DNA , Feminino , Expressão Gênica , Aconselhamento Genético , Glucosidases/genética , Humanos , Infertilidade Masculina/diagnóstico , Infertilidade Masculina/etnologia , Infertilidade Masculina/patologia , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase Multiplex , Aceitação pelo Paciente de Cuidados de Saúde , Rim Policístico Autossômico Dominante/diagnóstico , Rim Policístico Autossômico Dominante/etnologia , Rim Policístico Autossômico Dominante/patologia , Técnicas de Reprodução Assistida , Análise do Sêmen
19.
Macromol Rapid Commun ; 38(21)2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28898505

RESUMO

This study reports a remarkably facile method to synthesize novel ionogels with imidazolium cycle crosslinks based on polyamidoamine (PAMAM) dendrimers via one-pot, modified Debus-Radziszewski reaction in ionic liquid 1-ethyl-3-methylimidazolium acetate ([EMIM][OAc]). High room temperature ionic conductivity (up to 6.8 mS cm-1 ) is achieved, and more remarkably, it can still exceed 1 mS cm-1 when the dendrimer content reached 70% because PAMAM dendrimers are completely amorphous with many cavities and the newly formed imidazolium crosslinks contains ions. The elastic modulus of these ionogels can exceed 106 Pa due to the newly-formed rigid imidazolium crosslinks. Crucially, these ionogels are robust gels even at temperatures up to 160 °C. Such novel ionogels with high ionic conductivity, tunable modulus, and flexibility are desirable for use in high-temperature flexible electrochemical devices.


Assuntos
Reagentes de Ligações Cruzadas/química , Dendrímeros/química , Géis/síntese química , Imidazóis/química , Líquidos Iônicos/química , Elasticidade , Condutividade Elétrica , Géis/química , Temperatura , Viscosidade
20.
Pestic Biochem Physiol ; 140: 1-8, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28755688

RESUMO

Acute toxicities (LD50s) of imidacloprid and clothianidin to Apis mellifera and A. cerana were investigated. Changing patterns of immune-related gene expressions and the activities of four enzymes between the two bee species were compared and analyzed after exposure to sublethal doses of insecticides. Results indicated that A. cerana was more sensitive to imidacloprid and clothianidin than A. mellifera. The acute oral LD50 values of imidacloprid and clothianidin for A. mellifera were 8.6 and 2.0ng/bee, respectively, whereas the corresponding values for A. cerana were 2.7 and 0.5ng/bee. The two bee species possessed distinct abilities to mount innate immune response against neonicotinoids. After 48h of imidacloprid treatment, carboxylesterase (CCE), prophenol oxidase (PPO), and acetylcholinesterase (AChE) activities were significantly downregulated in A. mellifera but were upregulated in A. cerana. Glutathione-S-transferase (GST) activity was significantly elevated in A. mellifera at 48h after exposure to imidacloprid, but no significant change was observed in A. cerana. AChE was downregulated in both bee species at three different time points during clothianidin exposure, and GST activities were upregulated in both species exposed to clothianidin. Different patterns of immune-related gene expression and enzymatic activities implied distinct detoxification and immune responses of A. cerana and A. mellifera to imidacloprid and clothianidin.


Assuntos
Abelhas/efeitos dos fármacos , Guanidinas/toxicidade , Inseticidas/toxicidade , Neonicotinoides/toxicidade , Nitrocompostos/toxicidade , Tiazóis/toxicidade , Animais , Guanidinas/química , Antígenos de Histocompatibilidade , Inseticidas/química , Neonicotinoides/química , Nitrocompostos/química , Especificidade da Espécie , Tiazóis/química
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