RESUMO
This study aimed to investigate the effect of supplementing 300 mg/kg tea polyphenols (TP) on yolk cholesterol content and production performance of laying hens during the egg-laying period. A total of 600 Roman laying hens aged 24 weeks were randomly divided into two dietary treatment groups. The feeding experiment lasted for 48 weeks. Layers fed basal diet supplemented with 0 (control group) and 300mg/kg TP (TP group) diet, respectively. The yolk cholesterol content, laying performance, and egg quality were determined at 28, 38, 48, 58, and 68 weeks of age. The yolk cholesterol content in the TP group was significantly decreased at 28-68 weeks of age (p<0.01), compared to the control group. There was a significant increase in laying rate in the TP group at 38 weeks of age (p<0.05), compared to the control group, while no significant differences during the other laying periods were obtained (p>0.05). The FCR significantly decreased in the TP group at 38 weeks of age whereas AEW significantly increased in the TP group at 58 weeks of age (p<0.05). Similarly, the eggshell thickness and eggshell strength in the TP group significantly increased (p<0.05), compared with the control group at 38 weeks of age. The albumen height and Haugh unit significantly increased at 28 weeks of age (p<0.05). In conclusion, the results showed that the diet supplemented with 300 mg/kg TP had positive effects on production performance of layers during the egg-laying period, and could lessen yolk cholesterol content significantly at 28-68 weeks of age.(AU)
Assuntos
Animais , Feminino , Galinhas/fisiologia , Suplementos Nutricionais/análise , Gema de Ovo/química , Colesterol/análise , Polifenóis/efeitos adversosRESUMO
microRNA (miR)-142-3p is implicated in malignancy and has been identified as a biomarker for aggressive and recurrent lung adenocarcinomas. This study aimed to evaluate the inhibitory effect of miR-142-3p on apoptosis and inflammation induced by bleomycin in MLE-12 cells. MLE-12 cells were first transfected either with miR-142-3p mimic or miR-142-3p inhibitor and then the cells were exposed to 50 µg/mL of bleomycin. Thereafter, cell viability, apoptosis and the expression of pro-inflammatory cytokines were assessed using CCK-8, flow cytometry, RT-PCR and western blot analyses. Cox-2, PI3K, AKT and mTOR expressions were detected by western blotting after bleomycin was administered together with NS-398 (an inhibitor of Cox-2). As a result, cell viability was significantly decreased, as well as apoptosis and the expression of IL-1 and TNF-α were remarkably increased after 50 and 100 µg/mL of bleomycin administration. miR-142-3p overexpression alleviated bleomycin-induced apoptosis and overproduction of these two pro-inflammatory cytokines, while miR-142-3p suppression exhibited completely opposite results. Up-regulation of Cox-2 and inactivation of PI3K/AKT/mTOR were found in bleomycin-pretreated cells, while these abnormal regulations were partially abolished by miR-142-3p overexpression and NS-398. In conclusion, this study demonstrated that miR-142-3p overexpression protected bleomycin-induced injury in lung epithelial MLE-12 cells, possibly via regulating Cox-2 expression and PI3K/AKT/mTOR signaling pathway. These findings provide evidence that miR-142-3p may be a therapeutic strategy for idiopathic pulmonary fibrosis (IPF) treatment.
Assuntos
Apoptose/efeitos dos fármacos , Bleomicina/farmacologia , Ciclo-Oxigenase 2/metabolismo , Regulação para Baixo/efeitos dos fármacos , Pulmão/citologia , MicroRNAs/metabolismo , Linhagem Celular , Humanos , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , TransfecçãoRESUMO
Doubling method is the technical barriers in maize haploid breeding. It was very important to establish the independent intellectual property rights for doubling method. In this experiment, the maize haploid inducer, TG15, was used for producing maternal haploids. Also, haploids were obtained from two kinds of maternal genotypes involved in the experiment, including high-oil type and common type. Significant differences were observed among offspring of various genotypes in the recovery of haploid fertilization. In 21 hybrid offspring haploids, the average powder rate was 8.28%, and the seed setting rate was 4.98%. The experimental results showed that when the hybrids were treated with 0.08% colchicine, the average powder rate and seed setting rate of offspring haploids were 35.53 and 20.30%, respectively, which were significantly higher than the hybrids with natural recovery ability. This study primarily established the doubling method of haploids called "bud seedling method" in China which was very practicably in maize doubled haploid breeding.
Assuntos
Zea mays/genética , Cromossomos de Plantas , Colchicina/farmacologia , Genes de Plantas , Genótipo , Haploidia , Inflorescência/fisiologia , Melhoramento Vegetal/métodos , Sementes/genética , Zea mays/efeitos dos fármacosRESUMO
microRNA (miR)-142-3p is implicated in malignancy and has been identified as a biomarker for aggressive and recurrent lung adenocarcinomas. This study aimed to evaluate the inhibitory effect of miR-142-3p on apoptosis and inflammation induced by bleomycin in MLE-12 cells. MLE-12 cells were first transfected either with miR-142-3p mimic or miR-142-3p inhibitor and then the cells were exposed to 50 μg/mL of bleomycin. Thereafter, cell viability, apoptosis and the expression of pro-inflammatory cytokines were assessed using CCK-8, flow cytometry, RT-PCR and western blot analyses. Cox-2, PI3K, AKT and mTOR expressions were detected by western blotting after bleomycin was administered together with NS-398 (an inhibitor of Cox-2). As a result, cell viability was significantly decreased, as well as apoptosis and the expression of IL-1 and TNF-α were remarkably increased after 50 and 100 μg/mL of bleomycin administration. miR-142-3p overexpression alleviated bleomycin-induced apoptosis and overproduction of these two pro-inflammatory cytokines, while miR-142-3p suppression exhibited completely opposite results. Up-regulation of Cox-2 and inactivation of PI3K/AKT/mTOR were found in bleomycin-pretreated cells, while these abnormal regulations were partially abolished by miR-142-3p overexpression and NS-398. In conclusion, this study demonstrated that miR-142-3p overexpression protected bleomycin-induced injury in lung epithelial MLE-12 cells, possibly via regulating Cox-2 expression and PI3K/AKT/mTOR signaling pathway. These findings provide evidence that miR-142-3p may be a therapeutic strategy for idiopathic pulmonary fibrosis (IPF) treatment.
Assuntos
Humanos , Bleomicina/farmacologia , Regulação para Baixo/efeitos dos fármacos , Apoptose/efeitos dos fármacos , MicroRNAs/metabolismo , Ciclo-Oxigenase 2/metabolismo , Pulmão/citologia , Transfecção , Linhagem Celular , Pulmão/efeitos dos fármacos , Pulmão/metabolismoRESUMO
Cotton is one of the most important natural fiber crops in the world. Its growth and yield is greatly limited by drought. A quantitative trait locus (QTL) analysis was therefore conducted to investigate the genetic basis of drought tolerance in cotton (Gossypium spp) using 188 F2:3 lines developed from an inter-specific cross between a wild cotton species, G. tomentosum, and an upland cotton, G. hirsutum (CRI-12). A genetic map was constructed using 1295 simple sequence repeat markers, which amplified 1342 loci, distributed on 26 chromosomes, covering 3328.24 cM. A field experiment was conducted in two consecutive years (2014 and 2015) and 11 morphological and physiological traits were recorded under water-limited (W1)/well-watered (W2) regimes at three growth stages (bud, flowering, and full boll). The traits measured included chlorophyll content, plant height, leaf area, leaf number, leaf fresh weight, leaf dry weight, boll weight, number of bolls per plant, and the number of fruiting branches. Sixty-seven and 35 QTLs were found under the W1 and W2 conditions, respectively. Of these, the majority exhibited partial dominance or over-dominance genetic effects for increasing the trait values. Four consistent QTLs were found under the W1 treatment on chromosomes 5, 8, 9, and 16, whereas no consistent QTL was found in W2. Thirteen QTL clusters were also identified on nine chromosomes (2, 3, 5, 6, 9, 14, 15, 16, and 21). These results will help to elucidate the genetic basis of drought tolerance in cotton.
Assuntos
Adaptação Biológica/genética , Mapeamento Cromossômico , Cruzamentos Genéticos , Gossypium/genética , Locos de Características Quantitativas , Estresse Fisiológico/genética , Cromossomos de Plantas , Secas , Marcadores Genéticos , Repetições de MicrossatélitesRESUMO
Doubled haploid (DH) technology, which is used for rapidly purifying genetic resources, is a key technology in modern maize breeding. The present study evaluated the tissue culture characteristics of maize haploid coleoptile sections, in order to provide a new way of haploid doubling. With 20 combinations of haploid coleoptile sections, obtained by hybridization within Reid, Tangsipingtou, and Term-tropical groups, as explants, we analyzed the induction and differentiation rate of callus, observed the number of root tip chromosomes in regenerated plants, and analyzed the pollen fertility. In addition, we used 47 SSR markers to analyze the genotypes of regenerated plants. The Reid and Tangsipingtou groups had significantly higher induction rates of haploid coleoptile callus compared to the Term-tropical group. Fifteen haploid plants were obtained which had 10 chromosomes in the root tips as assessed by I-KI staining. It was also noticed that the pollen of pollinated anthers were partially fertile. The haploid plants had genetic stability and showed no variation. The Reid and Tangsipingtou groups had good culture characteristics of haploid coleoptile sections, while the Term-tropical group had poor culture characteristics. Genotypes of haploid plants generated by tissue culture were evidenced to come from recombinant types of parents. Thus, this study established a tissue culture system of maize haploid coleoptile.
Assuntos
Cotilédone/genética , Haploidia , Zea mays/genética , Cromossomos de Plantas , Genômica , Genótipo , Hibridização Genética , Repetições de Microssatélites , Fenótipo , Pólen , Regeneração , Técnicas de Cultura de TecidosRESUMO
We investigated weak cation magnetic separation technology and matrix-assisted laser desorption ionization-time of flight-mass spectrometry (MALDI-TOF-MS) in screening serum protein markers of primary type I osteoporosis. We selected 16 postmenopausal women with osteoporosis and nine postmenopausal women as controls to find a new method for screening biomarkers and establishing a diagnostic model for primary type I osteoporosis. Serum samples were obtained from controls and patients. Serum protein was extracted with the WCX protein chip system; protein fingerprints were examined using MALDI-TOF-MS. The preprocessed and model construction data were handled by the ProteinChip system. The diagnostic models were established using a genetic arithmetic model combined with a support vector machine (SVM). The SVM model with the highest Youden index was selected. Combinations with the highest accuracy in distinguishing different groups of data were selected as potential biomarkers. From the two groups of serum proteins, 123 cumulative MS protein peaks were selected. Significant intensity differences in the protein peaks of 16 postmenopausal women with osteoporosis were screened. The difference in Youden index between the four groups of protein peaks showed that the highest peaks had mass-to-charge ratios of 8909.047, 8690.658, 13745.48, and 15114.52. A diagnosis model was established with these four markers as the candidates, and the model specificity and sensitivity were found to be 100%. Two groups of specimens in the SVM results on the scatterplot were distinguishable. We established a diagnosis model, and provided a new serological method for screening and diagnosis of osteoporosis with high sensitivity and specificity.
Assuntos
Biomarcadores/sangue , Proteínas Sanguíneas/metabolismo , Cátions/administração & dosagem , Osteoporose Pós-Menopausa/sangue , Estudos de Casos e Controles , Feminino , Humanos , Magnetismo/métodos , Pessoa de Meia-Idade , Mapeamento de Peptídeos/métodos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodosRESUMO
Studies have shown that eosinophils are closely related to pathogenesis of bronchial asthma. Eosinophils release eosinophil cationic protein (ECP), which plays an important role in infection and allergic reactions. Serum ECP mRNA expression in children with bronchial asthma has not been adequately investigated. We analyzed serum ECP mRNA expression in 63 children with bronchial asthma and 21 healthy children by using reverse-transcriptase polymerase chain reaction to understand the role of ECP in children with bronchial asthma. The children with bronchial asthma were segregated into acute-phase and stable-phase groups, based on the severity of the illness. Serum ECP mRNA expression in children with bronchial asthma (0.375 ± 0.04) was significantly higher than that in healthy controls (0.20 ± 0.02; P < 0.05). Additionally, children in the acute-phase group showed higher ECP mRNA expression level (0.44 ± 0.06) than those in the stable-phase (0.31 ± 0.03) and healthy control groups (0.20 ± 0.02; P < 0.05), while the level in the stable-phase (0.31 ± 0.03) was markedly higher than that in the healthy control group (0.20 ± 0.02; P < 0.05). Detection of serum ECP mRNA expression level has possible applications in the diagnosis and treatment of children with bronchial asthma.
Assuntos
Asma/genética , Proteína Catiônica de Eosinófilo/genética , Eosinófilos/enzimologia , RNA Mensageiro/biossíntese , Asma/sangue , Asma/enzimologia , Hiper-Reatividade Brônquica/sangue , Hiper-Reatividade Brônquica/genética , Criança , Proteína Catiônica de Eosinófilo/biossíntese , Proteína Catiônica de Eosinófilo/sangue , Feminino , Humanos , Masculino , RNA Mensageiro/sangue , RNA Mensageiro/genéticaRESUMO
Nuclear factor-erythroid 2-related factor 2 (Nrf2) is a transcription factor that regulates antioxidant and anti-inflammatory genes, and it plays a crucial role in the pathogenesis of chronic obstructive pulmonary disease (COPD). Moreover, 15-deoxy-delta12,14-prostaglandin J2 (15d-PGJ2) plays a protective role against oxidative stress and inflammation both in vivo and in vitro. In a previous study, we found that 15d-PGJ2 increased the expression of Nrf2 in a COPD rat model. This study aims to elucidate the role of 15d-PGJ2 in COPD pathogenesis and the relationship between Nrf2 and human bronchial epithelial (HBE) cells. Normal HBE (HBE) cells were cultured. Following cigarette smoke extract (CSE) stimulation, pre-incubation with or without small interfering RNA (siRNA) Nrf2, and stimulation with or without 15d-PGJ2, the expression levels of Nrf2, NF-κBp65, and IL-8 were detected by reverse transcription-polymerase chain reaction and western blot, respectively. The expression of NF-κBp65 and IL-8 in CSE-stimulated normal HBE cells was inhibited by 15d-PGJ2 at both the mRNA level and the protein level. Moreover, the expression of Nrf2 in normal HBE cells was improved by 15d-PGJ2 at both the mRNA level and the protein level. However, the inhibitory or improving effects of 15d-PGJ2 were disengaged by siRNA Nrf2 at both the mRNA level and the protein level. 15d-PGJ2 possesses anti-inflammatory properties in the pathogenesis of COPD, and HBE cells stimulated by CSE via Nrf2 activation.
Assuntos
Anti-Inflamatórios/farmacologia , Fator 2 Relacionado a NF-E2/metabolismo , Prostaglandina D2/análogos & derivados , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Doença Pulmonar Obstrutiva Crônica/metabolismo , Animais , Brônquios/citologia , Brônquios/efeitos dos fármacos , Brônquios/metabolismo , Células Cultivadas , Células Epiteliais/citologia , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Humanos , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Masculino , Modelos Animais , Fator 2 Relacionado a NF-E2/antagonistas & inibidores , Fator 2 Relacionado a NF-E2/biossíntese , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/fisiologia , Prostaglandina D2/farmacologia , RNA Mensageiro/metabolismo , Ratos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais/efeitos dos fármacos , Fator de Transcrição RelA/antagonistas & inibidores , Fator de Transcrição RelA/biossíntese , Fator de Transcrição RelA/metabolismoRESUMO
The transfer of agronomically useful genes from wild wheat species into cultivated wheat is one of the most effective approaches to improvement of wheat varieties. To evaluate the transfer of genes from Dasypyrum villosum into Triticum aestivum, wheat quality and disease resistance was evaluated in two new translocation lines, T1DLâ¢1V#3S and T1DSâ¢1V#3L. We examined the levels of stripe rust resistance and dough quality in the two lines, and identified and located the stripe rust resistant genes and high molecular weight glutenin subunit (HMW-GS) genes Glu-V1 of D. villosum. Compared to the Chinese Spring (CS) variety, T1DLâ¢1V#3S plants showed moderate resistance to moderate susceptibility to the stripe rust races CYR33 and Su11-4. However, T1DSâ¢1V#3L plants showed high resistance or immunity to these stripe rusts. The genes for resistance to stripe rust were located on 1VL of D. villosum. In comparison to CS, the dough from T1DSâ¢1V#3L had a significantly shorter developing time (1.45 min) and stable time (1.0 min), a higher weakness in gluten strength (208.5 FU), and a lower farinograph quality index (18). T1DLâ¢1V#3S had a significantly longer developing time (4.2 min) and stable time (5.25 min), a lower weakness in gluten strength (53 FU) and a higher farinograph quality index (78.5). We also found that T1DSâ¢1V#3L had reduced gluten strength and dough quality compared to CS, but T1DLâ¢1V#3S had increased gluten strength and dough quality. The results of SDS-PAGE analysis indicated that Glu-V1 of D. villosum was located on short arm 1VS and long arm 1VL. These results prove that the new translocation lines, T1DSâ¢1V#3L and T1DSâ¢1V#3L, have valuable stripe rust resistance and dough quality traits that will be important for improving wheat quality and resistance in future wheat breeding programs.
Assuntos
Basidiomycota/fisiologia , Resistência à Doença/genética , Farinha/normas , Genes de Plantas , Glutens/genética , Doenças das Plantas/microbiologia , Poaceae/genética , Triticum/genética , Ecótipo , Eletroforese em Gel de Poliacrilamida , Doenças das Plantas/genética , Subunidades Proteicas/genéticaRESUMO
Hemophagocytic lymphohistiocytosis (HLH) is a life-threatening syndrome involving a final common pathway of hypercytokinemia, in which tumor necrosis factor (TNF)-α, interferon (IFN)-γ, and soluble interleukin 2-receptor-alpha (sIL-2Rα) are the key cytokines. Pre-B-cell colony-enhancing factor (PBEF) is an inflammatory cytokine involved in several inflammatory diseases. However, its role in HLH is unknown. In this study, we examined the role of PBEF in HLH. Plasma was collected from 22 children with HLH and 14 healthy children. The concentrations of plasma PBEF, TNF-α, IFN-γ, and sIL-2Rα were determined using an enzyme-linked immunosorbent assay. All clinical data were derived from medical records. In the acute phase, children with HLH had much higher PBEF, TNF-α, IFN-γ, and sIL- 2Rα levels than did healthy children (P < 0.05). After treatment, 13 HLH children improved and PBEF, TNF-α, and IFN-γ levels decreased to normal levels (P < 0.05); sIL-2Rα levels also decreased (P < 0.05), but remained above the normal level (P < 0.05). Two patients were lost to follow-up, while 7 patients showed a bad response to therapy and eventually died, showing high PBEF levels above those of the survivors (P < 0.01). PBEF level was significantly positively correlated with TNF-α, IFN-γ, sIL-2Rα, serum ferritin, and triglycerides (all P < 0.05), and was negatively correlated with fibrin (P < 0.05). PBEF appears to be involved in the inflammatory process of HLH, and elevated PBEF is related to disease activity. We are currently evaluating the role of PBEF as a marker for the diagnosis and management of patients.
Assuntos
Biomarcadores/sangue , Citocinas/sangue , Inflamação/sangue , Linfo-Histiocitose Hemofagocítica/sangue , Nicotinamida Fosforribosiltransferase/sangue , Pré-Escolar , Feminino , Humanos , Lactente , Inflamação/patologia , Interferon gama/sangue , Subunidade alfa de Receptor de Interleucina-2/sangue , Interleucina-6/sangue , Linfo-Histiocitose Hemofagocítica/patologia , Masculino , Fator de Necrose Tumoral alfa/sangueRESUMO
The gene for the nucleocapsid (N) protein of canine distemper virus was cloned into the pMD-18T vector, and positive recombinant plasmids were obtained by enzyme digestion and sequencing. After digestion by both EcoRI and KpnI, the plasmid was directionally cloned into the eukaryotic expression vector pcDNA; the positive clone pcDNA-N was screened by electrophoresis and then transfected into COS-7 cells. Immunofluorescence analysis results showed that the canine distemper virus N protein was expressed in the cytoplasm of transfected COS-7 cells. After emulsification in Freund's adjuvant, the recombinant plasmid pcDNA-N was injected into the abdominal cavity of 8-week-old BABL/c mice, with the pcDNA original vector used as a negative control. Mice were immunized 3 times every 2 weeks. The blood of immunized mice was drawn 2 weeks after completing the immunizations to measure titer levels. The antibody titer in the pcDNA-N test was 10(1.62 ± 0.164), while in the control group this value was 10(0.52 ± 0.56), indicating that specific humoral immunity was induced in canine distemper virus nucleocapsid protein-immunized mice.
Assuntos
Vírus da Cinomose Canina/genética , Vírus da Cinomose Canina/imunologia , Animais , Anticorpos Antivirais/sangue , Clonagem Molecular , Cães , Feminino , Imunização/veterinária , Camundongos , Camundongos Endogâmicos BALB C , Proteínas do Nucleocapsídeo/genética , Vacinas Virais/imunologiaRESUMO
This study aimed to determine the relationship between changes in the serum levels of macrophage migratory inhibitory factor (MIF), interleukin (IL) 17, and IL-10 during chronic hepatitis B treatment via Baraclude(®) (Bristol-Meyers Squibb). Thirty-six patients with chronic hepatitis B and 24 healthy individuals were selected as the experimental and control groups, respectively, and the serum levels of MIF, IL-17, and IL-10 were measured during the period in which the experimental group was treated with oral Baraclude(®); meanwhile, the alanine aminotransferase (ALT), hepatitis B virus (HBV) DNA, and HBV marker (M) levels were measured in the experimental group. In the experimental group, the ALT and HBV-DNA levels began to exhibit obvious decreases in week 4, and the MIF and IL-17 levels exhibited obvious increases in week 4 followed by gradual decreases; however, the IL-10 level exhibited an obvious decrease in week 12 and then increased gradually. These changes were significant when compared with the control group (P < 0.05). In conclusion, Baraclude(®) treatment not only actively suppressed HBV but also indirectly balanced the MIF, IL-17, and IL-10 levels and reduced the liver inflammatory response.
Assuntos
Antivirais/administração & dosagem , Guanina/análogos & derivados , Hepatite B Crônica/sangue , Interleucina-10/sangue , Interleucina-17/sangue , Oxirredutases Intramoleculares/sangue , Fatores Inibidores da Migração de Macrófagos/sangue , Administração Oral , Adolescente , Adulto , Alanina Transaminase/sangue , Estudos de Casos e Controles , Feminino , Guanina/administração & dosagem , Antígenos de Superfície da Hepatite B/sangue , Hepatite B Crônica/tratamento farmacológico , Humanos , Masculino , Pessoa de Meia-Idade , Resultado do Tratamento , Adulto JovemRESUMO
We examined the association between the methionine synthase reductase (MTRR A66G), methylenetetrahydrofolate reductase (MTHFR C677T and A1298C), and methionine synthase (MS A2756G) genotypes and non-obstructive male infertility in a Chinese population. This case-control study included 162 infertile Chinese patients with azoospermia (N = 100) or oligoasthenozoospermia (N = 62) and 120 fertile men as controls. The polymorphisms MTRR A66G, MTHFR C677T, A1298C, and MS A2756G were identified by direct DNA sequencing and the results were statistically analyzed. We found no association between the incidence of any of these variants in azoospermia patients and control populations. The frequency of the MTRR66 polymorphic genotypes (AG, AG+GG) was significantly higher in the oligoasthenozoospermia group compared to the controls (P = 0.013, 0.012). Our findings revealed an association between the single-nucleotide polymorphism A66G in the MTRR gene and male infertility, particularly in oligoasthenozoospermia males, suggesting that this polymorphism is a genetic risk factor for male infertility in Chinese men.
Assuntos
Ferredoxina-NADP Redutase/genética , Predisposição Genética para Doença/genética , Infertilidade Masculina/genética , Polimorfismo de Nucleotídeo Único , Alelos , Povo Asiático/genética , Azoospermia/etnologia , Azoospermia/genética , Sequência de Bases , Estudos de Casos e Controles , China , Análise Mutacional de DNA , Frequência do Gene , Predisposição Genética para Doença/etnologia , Genótipo , Humanos , Infertilidade Masculina/etnologia , MasculinoRESUMO
Genetic regulatory networks are the key to understanding biochemical systems. One condition of the genetic regulatory network under different living environments can be modeled as a synchronous Boolean network. The attractors of these Boolean networks will help biologists to identify determinant and stable factors. Existing methods identify attractors based on a random initial state or the entire state simultaneously. They cannot identify the fixed length attractors directly. The complexity of including time increases exponentially with respect to the attractor number and length of attractors. This study used the bounded model checking to quickly locate fixed length attractors. Based on the SAT solver, we propose a new algorithm for efficiently computing the fixed length attractors, which is more suitable for large Boolean networks and numerous attractors' networks. After comparison using the tool BooleNet, empirical experiments involving biochemical systems demonstrated the feasibility and efficiency of our approach.
Assuntos
Algoritmos , Fenômenos Bioquímicos/genética , Redes Reguladoras de Genes/genética , Computação Matemática , Simulação por Computador , Modelos GenéticosRESUMO
The ectopic expression of cellulase in biomass can reduce the cost of biofuel conversion. This trait modification technique is highly beneficial for biofuel production. In this study, we isolated an endo-1,4-beta-glucanase gene (EGV) from Trichoderma reesei and inserted this gene downstream of a fragment encoding the signal peptide Apo-SP in a modified pCAMBIA1301 vector to obtain an Apo-SP and AsRed fusion protein. Transient expression of this fusion protein in onion epidermal cells showed that the Apo-SP signal was localized to the plastids. EGV transgenic rice plants that did not carry screening marker genes were obtained through overexpression of the pDTB double T-DNA vector. Western blotting showed that EGV was expressed in the dry straw of T0 generation transgenic rice plants and in fresh leaves of the T1 generation. More importantly, our results also showed that the peptide product of EGV in the transgenic plants folded correctly and was capable of digesting the cellulase substrate CMC. Additionally, cellulase activity remained stable in the straw that had been dried at room temperature for three months. This study presents an important technical approach for the development of transgenic rice straw that has stable cellulase activity and can be used for biofuel conversion.
Assuntos
Celulase/biossíntese , Celulase/genética , Oryza/genética , Plantas Geneticamente Modificadas/genética , Biocombustíveis , DNA Bacteriano/genética , Regulação da Expressão Gênica de Plantas , Trichoderma/enzimologia , Trichoderma/genéticaRESUMO
Accumulating evidence has indicated the importance of cancer stem cells in carcinogenesis. The goal of the present study was to determine the effect of low-dose cisplatin on enriched liver cancer stem cells (LCSCs). Human hepatoblastoma HepG2 cells were treated with concentrations of cisplatin ranging from 1 to 5 μg/mL. Cell survival and proliferation were evaluated using a tetrazolium dye (MTT) assay. LCSCs were identified using specific markers, namely aldehyde dehydrogenase-1 (ALDH1) and CD133. The percentage of ALDH1+ or CD133+ cells was examined by flow cytometric analysis. The expression of ALDH1 and/or CD133 in HepG2 cells was determined by immunocytochemical analysis. Low-dose cisplatin treatment significantly decreased cell survival in HepG2 cells after 24 or 72 h. However, the percentage of LCSCs in the surviving cells was greatly increased. The percentage of ALDH1+ or CD133+ cells was increased in a time- and dose-dependent manner after treatment with 1-4 μg/mL cisplatin, whereas 5 μg/mL cisplatin exposure slightly reduced the number of positive cells. These findings indicate that low-dose cisplatin treatment may efficiently enrich the LCSC population in HepG2 cells.
Assuntos
Humanos , Antineoplásicos/administração & dosagem , Proliferação de Células/efeitos dos fármacos , Cisplatino/administração & dosagem , Hepatoblastoma/tratamento farmacológico , Neoplasias Hepáticas/tratamento farmacológico , Células-Tronco Neoplásicas/efeitos dos fármacos , Antígenos CD/análise , Linhagem Celular Tumoral , Carcinogênese/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cisplatino/uso terapêutico , Citometria de Fluxo , Glicoproteínas/análise , Hepatoblastoma/patologia , Imuno-Histoquímica , Isoenzimas/análise , Neoplasias Hepáticas/patologia , Células-Tronco Neoplásicas/citologia , Peptídeos/análise , Retinal Desidrogenase/análise , Sais de Tetrazólio , Biomarcadores Tumorais/análiseRESUMO
Manual cultivar identification diagram is a new strategy for plant cultivar identification based on DNA markers, providing information to efficiently separate cultivars. We tested 25 pairs of apple EST-SSR primers for amplification of PCR products from loquat cultivars. These EST-SSR primers provided clear amplification products from the loquat cultivars, with a relatively high transferability rate of 84% to loquat; 11 pairs of primers amplified polymorphic products. After analysis of 24 red-fleshed loquat accessions, we found that only 7 pairs of primers could clearly separate all of them. A cultivar identification diagram of the 24 cultivars was constructed using polymorphic bands from the DNA fingerprints and EST-SSR primers. Any two of the 24 cultivars could be rapidly separated from each other, according to the polymorphic bands from the cultivars; the corresponding primers were marked in the correct position on the cultivar identification diagram. This red-flesh loquat cultivar identification diagram can separate the 24 red-flesh loquat cultivars, which is of benefit for loquat cultivar identification for germplasm management and breeding programs.
Assuntos
Cruzamento , Eriobotrya/genética , Repetições de Microssatélites/genética , DNA de Plantas , Eriobotrya/crescimento & desenvolvimento , Marcadores Genéticos/genética , Malus/genéticaRESUMO
Accumulating evidence has indicated the importance of cancer stem cells in carcinogenesis. The goal of the present study was to determine the effect of low-dose cisplatin on enriched liver cancer stem cells (LCSCs). Human hepatoblastoma HepG2 cells were treated with concentrations of cisplatin ranging from 1 to 5 µg/mL. Cell survival and proliferation were evaluated using a tetrazolium dye (MTT) assay. LCSCs were identified using specific markers, namely aldehyde dehydrogenase-1 (ALDH1) and CD133. The percentage of ALDH1+ or CD133+ cells was examined by flow cytometric analysis. The expression of ALDH1 and/or CD133 in HepG2 cells was determined by immunocytochemical analysis. Low-dose cisplatin treatment significantly decreased cell survival in HepG2 cells after 24 or 72 h. However, the percentage of LCSCs in the surviving cells was greatly increased. The percentage of ALDH1+ or CD133+ cells was increased in a time- and dose-dependent manner after treatment with 1-4 µg/mL cisplatin, whereas 5 µg/mL cisplatin exposure slightly reduced the number of positive cells. These findings indicate that low-dose cisplatin treatment may efficiently enrich the LCSC population in HepG2 cells.
Assuntos
Antineoplásicos/administração & dosagem , Proliferação de Células/efeitos dos fármacos , Cisplatino/administração & dosagem , Hepatoblastoma/tratamento farmacológico , Neoplasias Hepáticas/tratamento farmacológico , Células-Tronco Neoplásicas/efeitos dos fármacos , Antígeno AC133 , Família Aldeído Desidrogenase 1 , Antígenos CD/análise , Biomarcadores Tumorais/análise , Carcinogênese/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Cisplatino/uso terapêutico , Citometria de Fluxo , Glicoproteínas/análise , Células Hep G2 , Hepatoblastoma/patologia , Humanos , Imuno-Histoquímica , Isoenzimas/análise , Neoplasias Hepáticas/patologia , Células-Tronco Neoplásicas/citologia , Peptídeos/análise , Retinal Desidrogenase/análise , Sais de TetrazólioRESUMO
To investigate the variance of exogenous gene expression driven by different promoters by in vivo electroporation, 3 plasmid vectors carrying different promoters were selected, and their driving strength was compared in developing chicken embryos. The 3 promoters included: 1) the CAG promoter (containing the cytomegalovirus (CMV) immediate early enhancer and the chicken ß-actin promoter), 2) the CMV promoter (the human CMV immediate early region enhancer), and 3) the SV40 promoter (Simian virus 40). The intensity of GFP expression driven by the 3 promoters was detected by fluorescence microscopy. The results clearly showed that the expression intensity of the reporter gene differed significantly among the 3 promoters. Chicken ß-actin promoter induced the highest intensity of GFP expression, while SV40 promoter induced the lowest intensity. Our results indicate that plasmids with appropriate promoters should be carefully selected to obtain strong exogenous gene expression by in vivo electroporation.