RESUMO
Objective: To investigate chromosome abnormality rate and related factors of spontaneous abortion in early pregnancy. Methods: A total of 831 tissue samples of spontaneous abortion in early pregnancy were collected from June 2015 to August 2018 in the First Affiliated Hospital of Nanjing Medical University. Chromosomal copy number was analyzed by next generation sequencing (NGS). The relationships between chromosome abnormality and maternal age, in vitro fertilization-embryo transfer (IVF-ET) pregnancy, number of previous spontaneous abortions, history of live birth were analyzed by statistical methods. Results: Among 831 tissue samples of spontaneous abortion in early pregnancy, 461 (55.5%, 461/831) were found to have chromosome abnormalities. Maternal age (OR=1.107, 95%CI: 1.070- 1.145) and history of live birth (OR=1.909, 95%CI: 1.182-3.083) were the positive correlative factors of chromosome abnormality. Times of previous spontaneous abortion (OR=0.807, 95%CI: 0.702-0.928) and IVF-ET pregnancy (OR=0.554, 95%CI: 0.404-0.760) were the negative correlative factors of chromosome abnormality. Conclusions: Chromosome abnormality is an important cause of spontaneous abortion in early pregnancy. The rate of chromosome abnormality increases with the increase of maternal age and the history of live birth, and decreases with the increase of number of previous spontaneous abortion and IVF-ET pregnancy.
Assuntos
Aborto Espontâneo , Transtornos Cromossômicos/genética , Transferência Embrionária , Fertilização in vitro , Aborto Espontâneo/genética , Aberrações Cromossômicas , Feminino , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Idade Materna , Gravidez , Taxa de Gravidez , Primeiro Trimestre da GravidezRESUMO
1. Melanoma differentiation-associated gene 5 (MDA5) is a critical member of cytosolic pattern recognition receptors (PRRs) that recognise viral RNA and mediate type I interferon secretion in host cells. 2. The objective of the present study was to identify and characterise the structure and expression of pigeon MDA5. 3. The full-length MDA5 cDNA was cloned from pigeon spleen using RT-PCR and RACE. The distribution and expression level of pigeon MDA5 in different tissues were determined by QRT-PCR. 4. The results showed that the full-length pigeon MDA5 cDNA had 3858 nucleotides (containing a 210-bp 5'-UTR, a 3030-bp open reading frame and a 618-bp 3'-UTR) encoding a polypeptide of 1009 amino acids. The deduced amino acid sequence contained six conserved structural domains typical of RIG-I-like receptor (RLR), including two tandem arranged N-terminal caspase activation and recruitment domains (CARDs), a DEAH/DEAD box helicase domain (DExDc), a helicase superfamily c-terminal domain (HELICc), a type III restriction enzyme (ResIII) and a C-terminal regulatory domain (RD). 5. The pigeon MDA5 showed 84.8%, 87.3%, 87.9% and 87.2% amino acid sequence identities with previously described homologues from chicken, duck, goose and Muscovy ducks, respectively, and phylogenetic analysis revealed a close relationship among these MDA5. 6. Pigeon MDA5 transcript was ubiquitously expressed in all seven tissues tested in healthy pigeons and showed a high level in the thymus gland and kidney. 7. These findings lay the foundation for further research on the function and mechanism of MDA5 in innate immune responses related to vaccinations and infectious diseases in the pigeon.
Assuntos
Proteínas Aviárias/genética , Columbidae/genética , Helicase IFIH1 Induzida por Interferon/genética , Sequência de Aminoácidos , Animais , Proteínas Aviárias/química , Proteínas Aviárias/metabolismo , Sequência de Bases , Columbidae/metabolismo , Perfilação da Expressão Gênica/veterinária , Helicase IFIH1 Induzida por Interferon/química , Helicase IFIH1 Induzida por Interferon/metabolismo , Filogenia , Alinhamento de Sequência/veterináriaRESUMO
Objective: To evaluate the efficiency of the application of array comparative genomic hybridization (array-CGH) in preimplantation genetic diagnosis or screening (PGD/PGS), and compare the clinical outcomes of different stage embryo biopsy. Methods: The outcomes of 381 PGD/PGS cycles referred in the First Affiliated Hospital of Nanjing Medical University from July 2011 to August 2015 were retrospectively analyzed. There were 320 PGD cycles with 156 cleavage-stage-biopsy cycles and 164 trophectoderm-biopsy cycles, 61 PGS cycles with 23 cleavage-stage-biopsy cycles and 38 trophectoderm-biopsy cycles. Chromosomal analysis was performed by array-CGH technology combined with whole genome amplification. Single embryo transfer was performed in all transfer cycles. Live birth rate was calculated as the main clinical outcomes. Results: The embryo diagnosis rate of PGD/PGS by array-CGH were 96.9%-99.1%. In PGD biopsy cycles, the live birth rate per embryo transfer cycle and live birth rate per embryo biopsy cycle were 50.0%(58/116) and 37.2%(58/156) in cleavage-stage-biopsy group, 67.5%(85/126) and 51.8%(85/164) in trophectoderm-biopsy group (both P<0.01). In PGS biopsy cycles, the live birth rate per embryo transfer cycle and live birth rate per embryo biopsy cycle were the same as 34.8%(8/23) in cleavage-stage-biopsy group, the same as 42.1%(16/38) in trophectoderm-biopsy group (both P>0.05). Conclusions: High diagnosis rate and idea live birth rate are achieved in PGD/PGS cycles based on array-CGH technology. The live birth rate of trophectoderm-biopsy group is significantly higher than that of cleavage-stage-biopsy group in PGD cycles; the efficiency of trophectoderm-biopsy is better.
Assuntos
Hibridização Genômica Comparativa , Implantação do Embrião/genética , Implantação do Embrião/fisiologia , Transferência Embrionária/métodos , Testes Genéticos , Diagnóstico Pré-Implantação/métodos , Análise de Sequência de DNA/métodos , Biópsia , Técnicas de Cultura Embrionária , Feminino , Humanos , Gravidez , Estudos Retrospectivos , Transferência de Embrião ÚnicoRESUMO
The present study was to investigate whether baicalin can prevent repeated exogenous corticosterone injection-induced depressive-like behaviors and explore its possible mechanisms. After a 21-day treatment with baicalin (10 and 20 mg/kg), sucrose preference in the sucrose preference test (SPT) and immobility time in forced swimming test (FST) were observed, serum corticosterone levels and brain-derived neurotrophic factor (BDNF) contents in the hippocampus were examined by enzyme-linked immunosorbent assay (ELISA). In addition, quantitative real-time polymerase chain reaction (qPCR) and western blot were used to detect the mRNA and protein expression in the hippocampus. The results showed that 21-day cortiscosterone injections caused depressive-like behaviors in mice, including the reduced sucrose preference and increased duration of immobility. Baicalin reversed these behavioral changes described above and restored serum corticosterone levels. Additionally, baicalin up-regulated the mRNA and protein expression of glucocorticoid receptor (GR) and BDNF, accompanied with the down-regulation of serum- and glucocorticoid-regulated kinase 1 (SGK1) in the hippocampus. Moreover, baicalin significantly increased the protein expression of 11ß-hydroxysteroid dehydrogenase-2 (11ß-HSD2) in the hippocampus. The present results confirmed the antidepressant-like effects of baicalin in a mice model of depression induced by corticosterone and suggested that its mechanism was possibly involved in reducing serum corticosterone and thereby increasing BDNF in the hippocampus.
Assuntos
Antidepressivos/farmacologia , Transtorno Depressivo/tratamento farmacológico , Flavonoides/farmacologia , Hipocampo/efeitos dos fármacos , Proteínas Imediatamente Precoces/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , 11-beta-Hidroxiesteroide Desidrogenase Tipo 2/metabolismo , Animais , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Corticosterona , Transtorno Depressivo/metabolismo , Sacarose Alimentar , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Comportamento Alimentar/efeitos dos fármacos , Comportamento Alimentar/fisiologia , Hipocampo/metabolismo , Atividade Motora/efeitos dos fármacos , Atividade Motora/fisiologia , RNA Mensageiro/metabolismo , Distribuição Aleatória , Receptores de Glucocorticoides/metabolismo , Natação , Percepção Gustatória/efeitos dos fármacos , Percepção Gustatória/fisiologiaRESUMO
In this study, chicken adipocytes were cultured to evaluate RNA interference by the leptin receptor gene. A small interfering RNA of the leptin receptor gene was synthesized, with a suppression rate of 60% being generated (P < 0.01). After the knockdown of the leptin receptor, the expression levels of certain genes decreased significantly; specifically, peroxisome proliferator-activated receptor γ, fatty acid synthase, adipose triglyceride lipase, and lipoprotein lipase. In addition, a significant increase in the expression of the adiponectin gene was documented. These results demonstrate that the leptin receptor gene might contribute to lipid metabolism by influencing the expressions of the peroxisome proliferator-activated receptor γ, fatty acid synthase, adipose triglyceride lipase, lipoprotein lipase, and adiponectin genes.
Assuntos
Adipócitos/metabolismo , Interferência de RNA , Receptores para Leptina/genética , Animais , Galinhas , Regulação da Expressão Gênica , Metabolismo dos Lipídeos/genética , RNA Interferente Pequeno/síntese química , RNA Interferente Pequeno/genética , TransfecçãoRESUMO
Several studies have documented the process of early embryonic development in poultry; however, the molecular mechanisms underlying its developmental regulation are poorly understood, particularly in ducks. In this study, we analyzed differential gene expression of embryos 6 and 25 h following oviposition to determine which genes regulate the early developmental stage in ducks. Among 216 randomly selected clones, 39 protein-encoding cDNAs that function in metabolism, transcription, transportation, proliferation/apoptosis, cell cycle, cell adhesion, and methylation were identified. Additionally, the full-length cDNA of the Nanog gene, encoding a 302-amino acid protein, was obtained. Quantitative real-time polymerase chain reaction analyses were performed to detect expression levels of the selected genes during early and late embryonic stages, which revealed that these genes are expressed in a particular spatial and temporal pattern. These results indicate that these genes may play pivotal roles in the process of area pellucida formation through a complex and precise regulatory network during development in duck embryos.
Assuntos
Patos/genética , Biblioteca Gênica , Sequência de Aminoácidos , Animais , Patos/embriologia , Patos/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Dados de Sequência MolecularRESUMO
Abundance of duck adipocyte fatty acid-binding protein (A-FABP) mRNA was detected in this study by quantitative real-time PCR. The results showed that duck A-FABP gene was expressed in muscular tissues and many organs, except pancreas, lung, and kidney, and highly expressed in adipose tissues, especially in sebum. The expression of A-FABP and adipocyte differentiation-related genes was upregulated by oleic acid, and the A-FABP knockdown suppressed the oleic acid-stimulated expression of these genes in the cultured duck adipocytes, indicating A-FABP might play an important role in duck adipocyte differentiation.
Assuntos
Adipócitos/citologia , Patos/fisiologia , Proteínas de Ligação a Ácido Graxo/metabolismo , Animais , Diferenciação Celular , Sobrevivência Celular , Proteínas de Ligação a Ácido Graxo/genética , Feminino , Expressão Gênica , Masculino , Ácido Oleico , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
1. The aim of this study was to identify the effect of perilla extract, a source of polyunsaturated fatty acids, on lipid metabolism and expression of lipid-related genes in livers of Shaoxing ducks. 2. Two hundred and forty 28-week-old laying ducks received a commercial diet with perilla extract added at 0 (control) or 200 mg/kg of feed. 3. Ducks fed on a diet with perilla extract had increased laying rates compared with control ducks. 4. Serum concentrations of triglycerides were reduced by perilla extract, while high-density lipoprotein cholesterol and total serum cholesterol increased. 5. The expression of genes involved in hepatic lipogenesis, sterol regulatory element-binding protein-1, acetyl CoA carboxylase, stearoyl CoA desaturase, fatty acid synthase, apolipoprotein B, and apolipoprotein very low density lipoprotein, were decreased in the perilla group. 6. The mRNA expression of peroxisome proliferators-activated receptor alpha and acyl-coenzyme A oxidase was enhanced following treatment with perilla extract, and a similar tendency was observed in the expression of liver fatty acid-binding protein. 7. The results show that a diet with 200 mg/kg perilla extract regulated fat metabolism of Shaoxing ducks by improving egg laying, altering serum lipid profiles, stimulating lipid catabolic gene expression and inhibiting lipogenic gene expression in the liver.
Assuntos
Patos/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Metabolismo dos Lipídeos , Fígado/efeitos dos fármacos , Perilla/química , Extratos Vegetais/farmacologia , Acil-CoA Oxidase/metabolismo , Ração Animal , Animais , Colesterol/sangue , HDL-Colesterol/sangue , Dieta/veterinária , Patos/metabolismo , Proteínas de Ligação a Ácido Graxo/genética , Proteínas de Ligação a Ácido Graxo/metabolismo , Feminino , Lipoproteínas/sangue , Fígado/fisiologia , PPAR alfa/genética , PPAR alfa/metabolismo , Reação em Cadeia da Polimerase , RNA Mensageiro/análise , RNA Mensageiro/genética , Reprodução/efeitos dos fármacos , Triglicerídeos/sangueRESUMO
Several studies have investigated that linoleic acid (LA) and eicosapentaenoic acid (EPA) affect cell proliferation and lipid catabolic gene expression in mammals. To determine if LA and EPA increase duck cell proliferation and lipid catabolic gene expression, the authors exposed duck primary hepatocyte cultures to LA or EPA. The results showed that both LA and EPA increased cell proliferation in a dose-dependent manner (100 µM). The effect on specific cell-cycle phases was also studied; LA and EPA (100 µM) deceased the proportion of cells in the G0/G1 phase from 83 to 80.8 and 80.3%, respectively, concomitant with an increase in the proportion of S-phase cells (11.5 and 10.5 vs. 8%, respectively). The expression of PPAR-α and PPAR-α target genes, such as acyl-CoA oxidase (ACOX), lipoprotein lipase (LPL), liver fatty acid-binding protein (L-FABP), was examined by quantitative real-time PCR. The results showed that the expression of the PPAR-α, ACOX, and LPL genes increased significantly following LA and EPA exposure, but that the expression of L-FABP remained unchanged. This study provides the first characterization of LA- and EPA-induced cell proliferation and PPAR-α and PPAR-α target gene transcriptional responses in duck primary hepatocyte cultures.