RESUMO
A growing body of research has demonstrated the tumor suppressive function of microRNA (miR)-1 in many cancers. Our study aimed to investigate its role in vestibular schwannoma (VS). We examined miR-1 expression in 95 VS specimens and 79 normal vestibular nerves using quantitative real-time polymerase chain reaction. Moreover, miR-1 mimics, miR-1 inhibitors, and negative control oligonucleotides were transfected into HEI-193 human VS cells to investigate the functional significance of miR-1 expression in this condition at a cellular level. Finally, the role of vascular endothelial growth factor A (VEGFA) in miR-1-mediated HEI-193 cell growth was confirmed. miR-1 levels were significantly reduced in VS specimens compared with normal vestibular nerve tissues (P < 0.001). In addition, low levels of miR-1 were associated with larger tumor volumes. In functional assays, miR-1 suppressed HEI-193 cell proliferation and colony formation, and enhanced apoptosis. VEGFA was verified as a target gene of miR-1, and VEGFA overexpression partially negated the effects of miR-1 on HEI-193 cells. These findings suggest that miR-1 suppresses VS growth by targeting VEGFA, and should be considered as a potential therapeutic target for treatment of this condition.
Assuntos
Apoptose/genética , Estudos de Associação Genética , Neuroma Acústico/genética , Neuroma Acústico/patologia , Fator A de Crescimento do Endotélio Vascular/metabolismo , Sequência de Bases , Linhagem Celular Tumoral , Proliferação de Células , Regulação Neoplásica da Expressão Gênica , Humanos , MicroRNAs , Ensaio Tumoral de Célula-TroncoRESUMO
We explored the effect of baculovirus P35 protein on apoptosis in the brain tissue of rats with acute cerebral infarction (ACI). A rat model of middle cerebral artery infarction was created. The rats were randomly divided into sham, model, and treatment groups. Baculovirus P35 protein was injected into the intracranial arteries of the treatment group rats. The rats in the model group were given an equal volume of phosphate-buffered saline. The rats were sacrificed after 72 h and the brain tissue was separated. The levels of caspase-3, Bcl-2, and Bax mRNA, the brain cell apoptosis index, and the infarct size were determined. After 72 h, the levels of caspase-3 and Bax mRNA in the model and treatment groups were significantly greater than in the sham group, and the levels of Bcl-2 mRNA were significantly smaller (P < 0.05). The levels of caspase-3 and Bax mRNA were significantly lower in the treatment group than in the model group, and the level of Bcl-2 mRNA was significantly greater (P < 0.05). Compared with the sham group, the brain tissue apoptosis index and the cerebral infarction area increased significantly in the model and treatment groups (P < 0.05). The brain tissue apoptosis index and cerebral infarction area in the treatment group were significantly lower than in the model group (P < 0.05). Baculovirus P35 protein can effectively inhibit brain cell apoptosis in rats with ACI. It delayed apoptosis and necrosis in subjects with ACI tissue and had a protective effect on brain tissue.
Assuntos
Apoptose/efeitos dos fármacos , Infarto Cerebral/metabolismo , Infarto Cerebral/patologia , Proteínas Virais/farmacologia , Animais , Caspase 3/genética , Caspase 3/metabolismo , Infarto Cerebral/tratamento farmacológico , Modelos Animais de Doenças , Expressão Gênica , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Proteínas Virais/administração & dosagem , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismoRESUMO
Crossing different ploidy grapes is an effective way to obtain new seedless cultivars. Although embryo rescue has been extensively applied in breeding seedless and triploid grapes, only a few improved cultivars have been developed. Based on preliminary studies, we set five crosses between tetraploid and diploid grape varieties to obtain new hybrid triploid germplasms. Additionally, we compared two different methods of performing in vitro embryo rescue and sowing in the development of hybrid triploid grape plants. The results showed that the germination rate of hybrid seeds was much lower (0-22.8%) than that of the self-pollinated seeds (50.9-61.2%) obtained though the same method of in vitro culture. Meanwhile, the seed germination rates of all crosses obtain through in vitro culture (0-61.2%) were higher than those obtained through sowing (0-42.0%). Identification of ploidy level confirmed that three lines obtained from the crosses of 'Ruby Seedless (2x) x Black Olympia (4x)' and 'Big black (2x) x Kyoho (4x)' were triploid, and one line from the cross of 'Big black (2x) x Kyoho (4x)' was haploid, and the others were diploid, tetraploid, or aneuploidy plants. Moreover, 4 haploid and 42 triploid surviving grape seedlings were planted in a vineyard after propagation. Therefore, an efficient system of breeding triploid seedless grapes using embryo rescue was established and 42 hybrid triploid germplasms were obtained for use in future studies.
Assuntos
Cruzamentos Genéticos , Diploide , Sementes , Vitis/genética , Cruzamento , Quimera , Genótipo , GerminaçãoRESUMO
The delta-like 2 homolog (DLK2) modulates adipogenesis, hematopoiesis, osteogenesis, and other cell-differentiation processes. In the present study, we detected potential polymorphisms in the DLK2 gene in 604 individuals of Qinchuan cattle by using PCR-RFLP and DNA-sequencing methods. Herein, we identified five novel single-nucleotide polymorphisms (SNPs) (g.888G>A, g.910A>G, g.995G>A, g.4321A>G, g.4850A>G) and analyzed their association with measured traits. Four of the five analyzed polymorphisms were associated with at least one of the following traits: body weight (BW), chest depth (CD), chest circumference (CC), back fat thickness (BT), and rib-eye area (REA). To the best of our knowledge, our research is the first to report the association of DLK2 gene polymorphisms with growth and meat quality traits in Qinchuan cattle. In summary, the results of our study suggest that the DLK2 gene can be used as a candidate gene in beef cattle breeding.
Assuntos
Estudos de Associação Genética , Peptídeos e Proteínas de Sinalização Intercelular/genética , Carne/normas , Polimorfismo Genético , Característica Quantitativa Herdável , Alelos , Animais , Bovinos , Feminino , Genética Populacional , Genótipo , Desequilíbrio de Ligação , Polimorfismo de Nucleotídeo Único , Análise de Sequência de DNARESUMO
The CDIPT is crucial to the fatty acid metabolic pathway, intracellular signal transduction and energy metabolism in eukaryotic cells. We detected three SNPs at 3'-untranslated regions (UTR), named 3'-UTR_108 A > G, 3'-UTR_448 G > A and 3'-UTR_477 C > G, of the CDIPT gene in 618 Qinchuan cattle using PCR-RFLP and DNA sequencing methods. At each of the three SNPs, we found three genotypes named as follows: AA, AB, BB (3'-UTR_108 A > G), CC, CD, DD (3'-UTR_448 G > A) and EE, EF, FF (3'-UTR_477 C > G.). Based on association analysis of these SNPs with ultrasound measurement traits, individuals of genotype BB had a significantly larger loin muscle area than genotype AA. Individuals of genotype CC had significantly thicker back fat than individuals of genotype DD. Individuals of genotype EE also had significantly thicker back fat than did individuals of genotype FF. We conclude that these SNPs of the CDIPT gene could be used as molecular markers for selecting and breeding beef cattle with superior body traits, depending on breeding goals.
Assuntos
Regiões 3' não Traduzidas/genética , CDP-Diacilglicerol-Inositol 3-Fosfatidiltransferase/genética , Bovinos/genética , Carne/normas , Polimorfismo de Nucleotídeo Único , Animais , Sequência de Bases , Bovinos/metabolismo , China , Gorduras/metabolismo , Frequência do Gene , Genótipo , Músculos/metabolismo , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Análise de Sequência de DNARESUMO
The aim of this study was to determine whether single nucleotide polymorphisms (SNPs) of bovine Dickkopf homolog 4 (DKK4) are associated with body measurement traits in Qinchuan cattle. By using PCR-SSCP technology and DNA sequencing, we discovered 5 DKK4 SNPs in Qingchuan cattle, including -65G>A and -77G>T in the 5'-untranslated region, 1532C>G and 1533T>C in exon 2, and 2088C>T in exon 3. The sequencing map showed that 1532C>G and 1533T>C were in close linkage disequilibrium and were treated as 1532C>G-1533T>C in this study. Allele frequencies were calculated and analyzed by the chi-square test, which showed that -65G>A and 1532C>G-1533T>C were in Hardy-Weinberg equilibrium (P > 0.05), whereas -77G>T and 2088C>T were not in all 633 tested Qinchuan cattle individuals (P < 0.01). Gene heterozygosity (HE), effective allele number (NE), and polymorphism information content (PIC) were 0.407, 1.686, and 0.324 at -65G>A; 0.472, 1.894, and 0.361 at -77G>T; 0.476, 1.908, and 0.363 at 1532C>G-1533T>C; and 0.218, 1.279, and 0.195 at 2088C>T. We also evaluated the potential association of these SNPs with body measurement traits in all 633 individuals; the results suggest that several SNPs in Qinchuan cattle DKK4 were significantly associated with body length, hip height, rump length, hip width, heart girth, and pin bone width (P < 0.05 and P < 0.01). These results suggest that bovine DKK4 could be used as candidate gene for Qinchuan cattle breeding.
Assuntos
Bovinos/genética , Peptídeos e Proteínas de Sinalização Intercelular/genética , Técnicas de Amplificação de Ácido Nucleico/veterinária , Animais , Sequência de Bases , Composição Corporal/genética , Peso Corporal/genética , Cruzamento , China , Feminino , Frequência do Gene , Marcadores Genéticos , Genótipo , Reação em Cadeia da Polimerase/veterinária , Polimorfismo de Nucleotídeo Único , Polimorfismo Conformacional de Fita Simples , Análise de Sequência de DNARESUMO
The aim of this study was to investigate the safety and efficacy of endovascular abdominal aortic aneurysm repair (EVAR) in nonagenarians. From May 2003 to March 2011, 12 nonagenarian abdominal aortic aneurysm patients were treated with endovascular repair including two emergencies. The mean age of patients was 92.5 ± 1.6 years (range: 90-95 years), and 11 of the patients (91.7%) were men. Technical success, perioperative complications, and mortality were evaluated. The follow-up protocol consisted of computed tomography angiograms or ultrasounds performed at 3, 6, 9, and 12 months, and annually thereafter. Seven patients were operated under general anesthesia and five under local anesthesia. There was 100% technical success with no need for open conversion. The endografts used included two Endurant endografts, four Talent endografts, and six Zenith endografts. The mean operative time was 3.4 ± 1.3 h, mean procedural blood loss was 150.5 ± 60.5 mL, and mean postoperative length of stay was 8.4 ± 2.3 days. Mortality rates were 8.3% at 30 days, 16.7% at 1 year, 41.7% at 3 years, and 75% at 5 years. The mean survival of the 11 patients who expired beyond the first 30 days was 28.5 months (range: 9-73 months). Overall, EVAR in nonagenarians was associated with acceptable procedural success and perioperative morbidity and mortality. The medium and long-term results suggested that EVAR may be of limited benefit in some patients who are aged >90 years. Therefore, individual patient selection is very important.