RESUMO
This study investigated the effect of calcium (Ca) and phytase interaction on growth performance and bone quality in 1-42-day-old broiler chickens. A total of 624 female one-day-old Ross 308 broilers were allotted to 13 treatments with four replicates and 12 birds per replicate. A 2 × 6 factorial experiment was designed to test the combinations of 0.50% and 1.00% Ca with 0, 500, 1,000, 2,500, 5,000, and 10,000 FTU/kg phytase in the basal diet (0.25% non-phytate phosphorus, NPP). The control diet contained adequate Ca and phosphorus (P). Dietary Ca, phytase, and their interaction affected growth performance and bone mineralization of broilers at 1-42 days of age (p<0.05). The broilers fed with 1.00% Ca had lower body weight gain (BWG) and feed intake (FI) compared with the birds fed with 0.50% Ca (p<0.05). The BWG, FI, leg bone weight, and ash weight of the broilers fed with 0.25% NPP were lower than those of birds fed with the control diet (p<0.05). The addition of 500-10,000 FTU/kg phytase improved growth rate and leg bone quality, especially at 1.00% Ca (p<0.05). No differences were observed in growth performance and bone quality of 42-day-old broilers fed with 1.00% Ca + 2,500-10,000 FTU/kg phytase and the control diet (p>0.05). These data indicated that high doses of phytase (2,500-10,000 FTU/kg) alleviate the negative effects of Ca and P imbalance (Ca-to-NPP ratio = 4.0) on growth performance and bone mineralization of broiler chickens.(AU)
Assuntos
Animais , Feminino , Calcificação Fisiológica/efeitos dos fármacos , Galinhas/fisiologia , Fósforo/análise , Cálcio da Dieta/análise , Fenômenos Fisiológicos da Nutrição AnimalRESUMO
Chronic myelomonocytic leukemia (CMML) is a hematologic malignancy that overlaps with myeloproliferative neoplasms (MPN) and myelodysplastic syndromes (MDS) and tends to transform into acute myeloid leukemia (AML). Among cases of CMML, > 90% have gene mutations, primarily involving TET2 (~ 60%), ASXL1 (~ 40%), SRSF2 (~ 50%), and the RAS pathways (~ 30%). These gene mutations are associated with both the clinical phenotypes and the prognosis of CMML, special CMML variants and pre-phases of CMML. Cytogenetic abnormalities and the size of genome are also associated with prognosis. Meanwhile, cases with ASXL1, DNMT3A, NRAS, SETBP1, CBL and RUNX1 mutations may have inferior prognoses, but only ASXL1 mutations were confirmed to be independent predictors of the patient outcome and were included in three prognostic models. Novel treatment targets related to the various gene mutations are emerging. Therefore, this review provides new insights to explore the correlations among gene mutations, clinical phenotypes, prognosis, and novel drugs in CMML.
Assuntos
Antineoplásicos/uso terapêutico , Leucemia Mielomonocítica Crônica/tratamento farmacológico , Leucemia Mielomonocítica Crônica/genética , Mutação , Proteínas de Transporte/genética , Aberrações Cromossômicas , Subunidade alfa 2 de Fator de Ligação ao Core/genética , Metilação de DNA , DNA Metiltransferase 3A/genética , Proteínas de Ligação a DNA/genética , Dioxigenases/genética , Epigênese Genética , Repressão Epigenética , GTP Fosfo-Hidrolases/genética , Genes ras , Tamanho do Genoma , Humanos , Leucemia Mieloide Aguda/genética , Leucemia Mielomonocítica Crônica/mortalidade , Proteínas de Membrana/genética , Síndromes Mielodisplásicas/genética , Proteínas Nucleares/genética , Fenótipo , Prognóstico , Proteínas Proto-Oncogênicas c-cbl/genética , Proteínas Repressoras/genética , Fatores de Processamento de Serina-Arginina/genética , Transdução de Sinais/genéticaRESUMO
BACKGROUND: Over the past few decades, immunological checkpoint therapy has been an increasingly prominent strategy in the treatment of tumors, including prostate cancer (PC). There are few systematic studies of the phenotypic of tumor-infiltrating immune cells in PC tissues. METHODS: CIBERSORT is an analytical tool for estimating the abundance of member cell types in mixed cell population by gene expression data. Herein, we analyzed different levels of tumor-infiltrating immunity cells in normal tissue compared with PC using CIBERSORT. RESULTS: The results showed that proportion of M1 macrophages and resting mast cells presented significant differences in prostate tumor than these normal tissues. A higher proportion of resting mast cells was associated with a worse outcome and M1 macrophages was associated with a favorable outcome. Moreover, the radiotherapy and targeted molecular therapy can affect the immune infiltration of M1 macrophages and resting mast cells. CONCLUSIONS: Resting mast cells and M1 macrophages has an important role in the prognosis of prostate cancer. Our data provides valuable information about the future treatment of PC.
Assuntos
Próstata/imunologia , Neoplasias da Próstata/imunologia , Imunidade Adaptativa , Humanos , Imunidade Inata , Linfócitos do Interstício Tumoral/metabolismo , Linfócitos do Interstício Tumoral/patologia , Macrófagos/metabolismo , Macrófagos/patologia , Masculino , Mastócitos/metabolismo , Mastócitos/patologia , Prognóstico , Neoplasias da Próstata/patologiaRESUMO
This study was undertaken to clarify the role and mechanism of pyruvate dehydrogenase kinase isoform 2 (PDK2) in chondrogenic differentiation of mesenchymal stem cells (MSCs). MSCs were isolated from femurs and tibias of Sprague-Dawley rats, weighing 300-400 g (5 females and 5 males). Overexpression and knockdown of PDK2 were transfected into MSCs and then cell viability, adhesion and migration were assessed. Additionally, the roles of aberrant PDK2 in chondrogenesis markers SRY-related high mobility group-box 6 (Sox6), type ΙΙ procollagen gene (COL2A1), cartilage oligomeric matrix protein (COMP), aggrecan (AGC1), type ΙX procollagen gene (COL9A2) and collagen type 1 alpha 1 (COL1A1) were measured by quantitative reverse-transcription polymerase chain reaction (qRT-PCR). The expressions of c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase (MAPK) and extracellular regulated protein kinase (ERK) were measured. Overexpressing PDK2 promoted cell viability, adhesion and inhibited cell migration in MSCs (all P<0.05). qRT-PCR assay showed a potent increase in the mRNA expressions of all chondrogenesis markers in response to overexpressing PDK2 (P<0.01 or P<0.05). PDK2 overexpression also induced a significant accumulation in mRNA and protein expressions of JNK, p38MAPK and ERK in MSCs compared to the control (P<0.01 or P<0.05). Meanwhile, silencing PDK2 exerted the opposite effects on MSCs. This study shows a preliminary positive role and potential mechanisms of PDK2 in chondrogenic differentiation of MSCs. It lays the theoretical groundwork for uncovering the functions of PDK2 and provides a promising basis for repairing cartilage lesions in osteoarthritis.
Assuntos
Condrogênese/fisiologia , Proteínas Quinases JNK Ativadas por Mitógeno/fisiologia , Sistema de Sinalização das MAP Quinases/fisiologia , Células-Tronco Mesenquimais/fisiologia , Proteínas Serina-Treonina Quinases/fisiologia , Fatores de Transcrição SOXE/fisiologia , Animais , Diferenciação Celular , Cadeia alfa 1 do Colágeno Tipo I , Feminino , Masculino , Piruvato Desidrogenase Quinase de Transferência de Acetil , Ratos , Ratos Sprague-Dawley , Ativação Transcricional , Regulação para CimaRESUMO
Simple sequence repeats (SSRs), one of the most powerful molecular markers, can be used for DNA fingerprinting, variety identification, genetic mapping, and marker-assisted selection. Using the pear's (Pyrus pyrifolia Nakai) 75,764 unigenes (55,676,271 bp) obtained by deep transcriptome sequencing, a total of 10,622 novel SSRs were identified in 9154 unigenes, accounting for 14.02% of all unigenes. The average length and distribution of these SSRs was about 16 bp and 5.24 kb, respectively. Dinucleotide repeat motifs were the main type, with a frequency of 55.87%, followed by trinucleotides (24.45%). There were 159 kinds of repeat motifs existing in the pear transcriptome. AG/CT was the most frequent motif, accounting for 49.64%. All 9154 SSR-containing unigenes were functionally annotated using Nr (NCBI non-redundant protein database), Nt (NCBI non-redundant nucleotide database), and the Swiss-Prot database, and were classified further by Gene Ontology and Clusters of Orthologous Groups. In addition, a total of 4300 primer pairs were designed from all SSR loci obtained. Of these, 40 primers were randomly selected for PCR amplification and polyacrylamide gel (PAGE) analysis. Among the 40 primer pairs, 31 were successfully separated via PAGE. These findings also confirm that mining SSRs using next-generating sequencing technologies is a fast, effective, and reliable approach.
Assuntos
Repetições de Microssatélites/genética , Anotação de Sequência Molecular , Pyrus/genética , Análise de Sequência de RNA/métodos , Transcriptoma/genética , Sequência de Bases , Primers do DNA/metabolismo , Eletroforese em Gel de Poliacrilamida , Motivos de Nucleotídeos/genética , Reprodutibilidade dos TestesRESUMO
The aim of this study was to evaluate the effects of miR-26a on Beclin 1 expression in retinoblastoma (RB) cell lines (Y79 and WERi-RB-1). RB cells were transfected with miR-26a mimic, antagomir-26a, or control mimic. The Beclin 1 mRNA and protein levels were detected by quantitative polymerase chain reaction and western blot, respectively. The activity of Beclin 1 3ê-UTR reporter gene was detected with the luciferase assay. After transfection with miR-26a mimic, Beclin 1 mRNA and protein levels as well as the activity of the 3'-UTR reporter gene decreased. However, all were increased upon inhibition of miR-26a with antagomir-26a. Beclin 1 is the target of miR-26a in human RB cell lines Y79 and WERi-RB-1, and miR-26a inhibits the expression of Beclin 1 by reducing its mRNA and protein levels.
Assuntos
Proteína Beclina-1/biossíntese , MicroRNAs/genética , Neoplasias da Retina/genética , Retinoblastoma/genética , Regiões 3' não Traduzidas , Proteína Beclina-1/antagonistas & inibidores , Proteína Beclina-1/genética , Linhagem Celular Tumoral , Proliferação de Células/genética , Humanos , MicroRNAs/antagonistas & inibidores , MicroRNAs/biossíntese , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Neoplasias da Retina/metabolismo , Neoplasias da Retina/patologia , Retinoblastoma/metabolismo , Retinoblastoma/patologia , TransfecçãoRESUMO
OBJECTIVE: To investigate the role of the vascular endothelial growth factor receptor 2 (VEGFR2) in the proliferation, migration, invasion, and radiation-induced apoptosis of the non-small cell lung cancer (NSCLC) cell line Calu-1. METHODS: VEGFR2 gene was silenced by RNA interference in Calu-1 cells, and the expression of VEGFR2 was measured by qRT-PCR and Western blot analysis. The cells were divided into control, VEGF-treated, VEGFR2 knockdown, and VEGFR2 knockdown and VEGF-treated groups. A CCK8 assay and Transwell assay were performed to assess cell proliferation, migration, and invasion, respectively, after VEGFR2 knockdown. Western blot assays were used to detect signaling proteins downstream of VEGFR2. Cells in the groups listed above were also subjected to radiation treatment, followed by apoptosis analysis. RESULTS: (1) RNA interference of VEGFR2 in Calu-1 cells reduced VEGFR2 mRNA (P < 0.01) and protein levels (P < 0.01). (2) VEGFR2 knockdown inhibited proliferation (P < 0.05), migration (P < 0.05), and invasion (P < 0.05) in Calu-1 cells. (3) VEGFR2 knockdown blocked the phosphorylation of protein kinase B (Akt, also known as PKB), extracellular regulated kinase (ERK) 1/2, and p38 mitogen-activated protein kinase (p38 MAPK) to various extent (P < 0.05), but did not change their total protein expression. (4) Knockdown of VEGFR2 suppressed HIF-1α protein synthesis (P < 0.05), and exacerbated apoptosis induced by radiation (P < 0.05). CONCLUSION: VEGFR2 gene knockdown significantly suppressed a number of cellular activities in Calu-1 cells and increased radiation-induced cell death.
Assuntos
Carcinoma de Células Escamosas/patologia , Movimento Celular , Proliferação de Células/genética , Neoplasias Pulmonares/patologia , Tolerância a Radiação/genética , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/antagonistas & inibidores , Apoptose/efeitos da radiação , Western Blotting , Linhagem Celular Tumoral , Citometria de Fluxo , Técnicas de Silenciamento de Genes , Humanos , Invasividade Neoplásica , RNA Interferente Pequeno , Reação em Cadeia da Polimerase em Tempo Real , Transfecção , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/genéticaRESUMO
BACKGROUND: To determine the effects of endostatin on vascular growth factor receptor 2 (VEGFR2) expression in non-small cell lung cancer (NSCLC) cells and the mechanisms underlying its radiosensitizing effect. METHODS: VEGFR2 mRNA levels were determined in different NSCLC cell lines using qRT-PCR. RT-PCR and Western blot assays were used to assess the expression of mRNA and proteins. The radiosensitivity of the cells was determined by colony-formation assays; and cell apoptosis and cell cycle distribution were determined by flow cytometry. RESULTS: VEGFR2 mRNA levels differed among the five NSCLC cell lines (P < 0.01), with the highest expression in Calu-1 cells and lowest in A549 cells. Endostatin significantly inhibited the growth of Calu-1 cells (P < 0.01) (IC20 = 296.5 µg/ml), and the expression of VEGFR2 and HIF-1α (P < 0.05). Phosphorylation of protein kinase B (Akt), extracellular signal-regulated kinases 1/2 (ERK1/2), and p38 were significantly lower in endostatin-treated cells than control (P < 0.05). Endostatin enhanced the radiosensitivity of Calu-1 cells to SER = 1.38 and induced apoptosis (P < 0.01) and G2/M blockage (P < 0.01). However, endostatin had limited effects on A549 cells. Compared with Calu-1 cells, there was not significantly effects on cell radiosensitivity (SER = 1.09). CONCLUSIONS: Endostatin induces apoptosis and enhances radiosensitivity of the VEGFR2 high-expressing cell line Calu-1, but it has a limited effect on the VEGFR2 low-expressing cell line A549.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/genética , Endostatinas/farmacologia , Neoplasias Pulmonares/genética , Radiossensibilizantes/farmacologia , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/genética , Carcinoma Pulmonar de Células não Pequenas/patologia , Morte Celular/efeitos dos fármacos , Morte Celular/genética , Linhagem Celular Tumoral , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/genética , Raios gama , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Neoplasias Pulmonares/patologia , Tolerância a Radiação/efeitos dos fármacos , Tolerância a Radiação/genéticaRESUMO
The aim of this study was to investigate the correlation between the A1166C polymorphism in the angiotensin II type 1 receptor (AT1R) gene and heart failure (HF) risk using metaanalysis. The PubMed database was searched, and data were extracted independently by two reviewers. Odds ratios (ORs) with corresponding 95% confidence intervals (CIs) were used to assess the strength of the associations. Statistical analysis was performed using the STATA 12.0 software. The results of the metaanalysis showed no significant association between the AT1R A1166C polymorphism and HF risk (AA vs CC: OR = 0.72, 95%CI = 0.31-1.68; AA vs AC: OR = 0.78, 95%CI = 0.52-1.18; dominant model: OR = 1.37, 95%CI = 0.92-2.04; recessive model: OR = 0.73, 95%CI = 0.30-1.75). In the subgroup analysis by ethnicity, the results also showed no significant association between A1166C polymorphism and susceptibility to HF in both Caucasian and Asian populations. In conclusion, this meta-analysis suggests that the A1166C polymorphism in AT1R may not be associated with susceptibility to HF. Further large and well-designed studies are needed to confirm these conclusions.
Assuntos
Estudos de Associação Genética , Predisposição Genética para Doença , Insuficiência Cardíaca/genética , Polimorfismo de Nucleotídeo Único , Receptor Tipo 1 de Angiotensina/genética , Alelos , Substituição de Aminoácidos , Estudos de Casos e Controles , Códon , Humanos , Razão de Chances , Viés de Publicação , RiscoRESUMO
We observed the variation in in vivo blood lipid and blood glucose metabolism in rats with atherosclerosis after 5-(3,4-dihydroxy-phenyl)-1-piperidin-1-yl-penta-2,4-dien-1-one (GBOT) administration. Wistar rats aged 10 weeks received a high-fat diet to establish the atherosclerosis model. Metabolic indices related to blood lipid and blood glucose were measured before modeling and at 4 and 8 weeks after modeling. Liver fat levels in rats were measured at 8 weeks to analyze the relationship between liver fat and blood lipid levels. We examined the mechanism of blood lipid reduction. The levels of serum triglycerides, total cholesterol, and very-low-density lipoprotein cholesterol in rats in the control group were significantly decreased (P < 0.05) compared with those in the 4-week control group at 4 weeks and decreased significantly and continuously until the 8th week (P < 0.05). Compared with the 8-week control group, the blood glucose level in rats in the 8-week experimental group decreased significantly (P < 0.05), and the level of insulin sensitivity index decreased significantly (P < 0.05). Compared with the control group, triglyceride and total cholesterol levels per unit mass in rat liver tissue in the 8-week experimental group decreased significantly (P < 0.05). Western blotting indicated that GBOT significantly increased the expression of lecithin-cholesterol acyltransferase, low-density lipoprotein receptor, and cholesterol 7 alpha-hydroxylase proteins. GBOT can significantly decrease the levels of blood lipid and blood glucose in rat models of atherosclerosis, and its mechanism may be associated with the promotion of expression of lecithin-cholesterol acyltransferase, low-density lipoprotein receptor, and cholesterol 7 alpha-hydroxylase proteins.