RESUMEN
This study investigated SNP mutation sites of Gonadotrophin releasing hormone (GnRH) gene in China yellow quail, Beijing white quail and Korean quail through PCR amplification and DNA sequencing technologies. Moreover, polymorphism of GnRH gene and its association with growth traits of quail were analyzed, aiming to get molecular markers associated to growth traits of quail, which could provide references for breeding of new quail species. According to research results, a total of 14 SNP mutation sites of GnRH were detected in China yellow quail, Beijing white quail and Korean quail, which were C71T, C108T, C168T, C178T, A184G, C206T, A209C, C215T, A252G, A279T, C281T, C293G, C339T and C458T. Except that only 2 genotypes were detected for A209C and C281T in China yellow quail and Beijing white quail, 3 genotypes were detected for all of the remaining 12 SNP mutation sites in three quail species. Of the 14 SNP sites, C71T, A209C, C215T, C281T, C293G, C339T and C458T were significantly associated with body weight (p 0.05), C71T, C108T, C168T, C178T, A184G, C206T, C215T, A252G, C293G, C339T and C458T were significantly associated with shank length (p 0.05), C71T, C215T, C293G and C458T were significantly associated with breastbone length (p 0.05), A209C and C281T were significantly associated with shank circumference (p 0.05).
Asunto(s)
Animales , Coturnix/crecimiento & desarrollo , Coturnix/fisiología , Gonadotropinas , Peso Corporal , Polimorfismo GenéticoRESUMEN
This study investigated SNP mutation sites of Gonadotrophin releasing hormone (GnRH) gene in China yellow quail, Beijing white quail and Korean quail through PCR amplification and DNA sequencing technologies. Moreover, polymorphism of GnRH gene and its association with growth traits of quail were analyzed, aiming to get molecular markers associated to growth traits of quail, which could provide references for breeding of new quail species. According to research results, a total of 14 SNP mutation sites of GnRH were detected in China yellow quail, Beijing white quail and Korean quail, which were C71T, C108T, C168T, C178T, A184G, C206T, A209C, C215T, A252G, A279T, C281T, C293G, C339T and C458T. Except that only 2 genotypes were detected for A209C and C281T in China yellow quail and Beijing white quail, 3 genotypes were detected for all of the remaining 12 SNP mutation sites in three quail species. Of the 14 SNP sites, C71T, A209C, C215T, C281T, C293G, C339T and C458T were significantly associated with body weight (p 0.05), C71T, C108T, C168T, C178T, A184G, C206T, C215T, A252G, C293G, C339T and C458T were significantly associated with shank length (p 0.05), C71T, C215T, C293G and C458T were significantly associated with breastbone length (p 0.05), A209C and C281T were significantly associated with shank circumference (p 0.05).(AU)
Asunto(s)
Animales , Coturnix/crecimiento & desarrollo , Coturnix/fisiología , Gonadotropinas , Polimorfismo Genético , Peso CorporalRESUMEN
PURPOSE: The purpose of this study was to explore the differences between stage T2N0M0 and stage T1N1M0 gastric cancer (GC) and to identify the necessity of adjuvant treatment (AT) for these stages. METHODS: Between years 2004 and 2015, 1971 stage IB GC patients who underwent radical surgery were recruited using the Surveillance, Epidemiology and End Results database. We conducted univariate/multivariate analyses, the propensity score matching and evaluated gastric cancer-specific survival (GCSS) and overall survival (OS) with the log-rank test. RESULTS: T1N1M0 had a significantly worse survival than T2N0M0 in both GCSS and OS before and after the propensity score matching. Examined lymph nodes (ELN) ≤ 15 and T1N1M0 were independent risk factors for worse GCSS and OS in stage IB GC. The absence of adjuvant chemotherapy (CT) was an independent risk factor for worse GCSS and OS in T1N1M0 but not in T2N0M0. AT demonstrated similar GCSS and OS with surgery alone (SA) for T2N0M0 but better survival for T1N1M0. Compared to CT and adjuvant chemoradiotherapy (CRT) group, SA demonstrated significantly worse GCSS and OS for T1N1M0. There was no significant difference between CT and CRT in both T2N0M0 and T1N1M0 stages. T2N0M0 had a better survival than T1N1M0 in ELN ≤ 15 subgroup. However, similar survival was demonstrated in ELN > 15 subgroup. CONCLUSIONS: T2N0M0 GC has a better survival rate than T1N1M0 GC when ELN are ≤ 15. Moreover, T2N0M0 GC may not benefit from AT. T1N1M0 GC requires CT but not adjuvant radiotherapy.
Asunto(s)
Ganglios Linfáticos/patología , Estadificación de Neoplasias , Neoplasias Gástricas/patología , Neoplasias Gástricas/terapia , Adolescente , Adulto , Anciano , Análisis de Varianza , Quimioradioterapia Adyuvante/mortalidad , Quimioterapia Adyuvante/mortalidad , Niño , Preescolar , Femenino , Humanos , Lactante , Recién Nacido , Estimación de Kaplan-Meier , Ganglios Linfáticos/cirugía , Masculino , Persona de Mediana Edad , Estadificación de Neoplasias/mortalidad , Puntaje de Propensión , Programa de VERF , Factores Socioeconómicos , Neoplasias Gástricas/mortalidad , Tasa de Supervivencia , Adulto JovenRESUMEN
The aim of this study was to characterize variations in Raf kinase inhibitor protein (RKIP) expression and related signaling molecules in gastric cardia adenocarcinoma. Cancerous and precancerous tissues were collected from patients with gastric cardia adenocarcinoma and normal tissue was collected from healthy controls. RKIP expression was detected in these tissues and the serum levels of NF-κB p65 and T-lymphocyte subsets were measured. Positive RKIP expression was higher in gastric cardia adenocarcinoma tissues than in precancerous tissues. The serum level of total NF-κB p65 was higher in patients with gastric cardia adenocarcinoma than in healthy controls. Levels of NF-κB p65 did not correlate with positive and negative expression of RKIP, but were higher in patients with lymph node metastasis than in those without it. The cellular immune function of the gastric cardia adenocarcinoma group was lower than in normal controls, particularly in cases with negative RKIP expression. RKIP is downregulated in gastric cardia adenocarcinoma tissues, which is related to the occurrence, progression, invasion, and metastasis of tumors. The possible mechanism for this may be the inhibition of NF-κB activity and cellular immune function, which allows for the escape of tumor cells from immune surveillance.
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Adenocarcinoma/inmunología , Adenocarcinoma/metabolismo , Proteínas de Unión a Fosfatidiletanolamina/metabolismo , Neoplasias Gástricas/inmunología , Neoplasias Gástricas/metabolismo , Subgrupos de Linfocitos T/inmunología , Factor de Transcripción ReIA/metabolismo , Adenocarcinoma/sangre , Adenocarcinoma/patología , Adulto , Anciano , Anciano de 80 o más Años , Biomarcadores , Progresión de la Enfermedad , Femenino , Expresión Génica , Humanos , Inmunohistoquímica , Inmunofenotipificación , Metástasis Linfática , Recuento de Linfocitos , Masculino , Persona de Mediana Edad , Estadificación de Neoplasias , Fenotipo , Proteínas de Unión a Fosfatidiletanolamina/genética , Neoplasias Gástricas/sangre , Neoplasias Gástricas/patología , Subgrupos de Linfocitos T/metabolismo , Factor de Transcripción ReIA/sangreRESUMEN
Aquaporin (AQP)-1 and AQP-4 expression in lung tissues of SD rats during high altitude hypoxic lung injury, and the relationship between AQP-1 and AQP-4 expression, and acute hypoxic lung injury was analyzed. Thirty six healthy SD rats were divided into hypoxia 1d, 2d, 3d, 5d, and 7d groups and control group (N = 6). Pathological changes in lung tissue were observed by hematoxylin and eosin staining; lung injury was scored, and ultrastructural changes in lung tissue were observed by transmission electron microscopy. Changes in moisture content in lung tissues were determined by analyzing the wet/dry weight ratio (W/D). Localization of AQP-1 and AQP-4 was determined by immunohistochemistry. AQP-1 and AQP-4 expression were detected by western blot. Lung W/D was lower in hypoxia groups than in control group, and the highest in 3d group (P < 0.05). Light microscopy revealed a thickening alveolar wall and outstretched and congestive alveolar wall in hypoxia group; electron microscopy revealed the presence of abnormal alveolar type II epithelial cells, cavitation in cytoplasm, microvillus-like protrusions, and a reduced lamellar body. AQP- 1 and AQP-4 were mainly distributed in the capillaries and lymphatic and alveolar epithelial cells and airway epithelial cells, respectively. AQP-1 protein expression was decreased (western blot) in hypoxia 1d group (the lowest in 3d group; P < 0.05); there were no significant changes about AQP- 4 expression. Therefore, AQP-1 may be involved in abnormal transport of liquid ALI and pathogenesis of lung edema. AQP-4 may not be involved in the formation of ALI lung edema.
Asunto(s)
Lesión Pulmonar Aguda/metabolismo , Acuaporina 1/metabolismo , Acuaporina 4/metabolismo , Pulmón/metabolismo , Lesión Pulmonar Aguda/patología , Animales , Pulmón/patología , RatasRESUMEN
Cyclin B is a regulatory subunit of maturation-promoting factor (MPF), which has a key role in the induction of meiotic maturation of oocytes. MPF has been studied in a wide variety of animal species; however, its expression in crustaceans is poorly characterized. In this study, the complete cDNA sequence of Cyclin B was cloned from the red claw crayfish, Cherax quadricarinatus, and its spatiotemporal expression profiles were analyzed. Cyclin B cDNA (1779 bp) encoded a 401 amino acid protein with a calculated molecular weight of 45.1 kDa. Quantitative real-time PCR demonstrated that Cyclin B mRNA was expressed mainly in the ovarian tissue and that the expression decreased as the ovaries developed. Immunofluorescence analysis revealed that the Cyclin B protein relocated from the cytoplasm to the nucleus during oogenesis. These findings suggest that Cyclin B plays an important role in gametogenesis and gonad development in C. quadricarinatus.
Asunto(s)
Astacoidea/genética , Ciclina B/genética , Regulación del Desarrollo de la Expresión Génica , Factor Promotor de Maduración/genética , Oocitos/metabolismo , Oogénesis/genética , Secuencia de Aminoácidos , Animales , Astacoidea/citología , Astacoidea/crecimiento & desarrollo , Secuencia de Bases , Núcleo Celular/metabolismo , Clonación Molecular , Ciclina B/metabolismo , Citoplasma/metabolismo , ADN Complementario/genética , ADN Complementario/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Femenino , Factor Promotor de Maduración/metabolismo , Meiosis , Datos de Secuencia Molecular , Peso Molecular , Oocitos/citología , Oocitos/crecimiento & desarrollo , Sistemas de Lectura Abierta , Ovario/citología , Ovario/crecimiento & desarrollo , Ovario/metabolismo , Transporte de Proteínas , Reacción en Cadena en Tiempo Real de la Polimerasa , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Alineación de SecuenciaRESUMEN
The insulin-like growth factor 2 receptor gene (IGF2R) encodes a transmembrane protein receptor and acts to sequester and degrade excess circulating insulin-like growth factor 2, which is critical for normal mammalian growth and development. Thus, IGF2R may serve as a candidate gene underlying growth trait in the common carp. In this study, we isolated the intron one of common carp IGF2R and detected the diversity in 3 continuous generations of FFRC strain common carp. A total of 8 loci were detected within this region, which were named in accordance with their location (i.e., Loc84, Loc106, Loc119, Loc130, Loc145, Loc163, Loc167, and Loc265). Loc106, Loc119, and Loc145 were moderately polymorphic; while Loc84, Loc130, Loc163, Loc167, and Loc265 exhibited slight level of polymorphism. However, significant differences between polymorphism information content values were not observed among the different generations. For Loc145, all generations deviated from Hardy-Weinberg equilibrium. The total number of significant linkage disequilibria for all generations equaled 40. Among them, 4 pairs were detected in each population, while 8 pairs were found in the 2nd and 3rd generations. For Loc130, the G/T genotype exhibited higher body weight when compared to that of the G/G genotype. The frequency of the homozygous G/G genotype reached 87.96%; thus, we can improve FFRC strain common carp growth performance by increasing the percentage of the G/T genotype within a breeding population. Therefore, the G/T genotype could be used as a molecular marker for superior growth traits.
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Carpas/crecimiento & desarrollo , Carpas/genética , Intrones/genética , Polimorfismo de Nucleótido Simple/genética , Receptor IGF Tipo 2/genética , Animales , Peso Corporal/genética , Sitios Genéticos , Heterocigoto , Desequilibrio de Ligamiento/genética , Filogenia , Reacción en Cadena de la PolimerasaRESUMEN
The Wnt inhibitor dickkopf-1 (DKK-1) has been shown to be closely correlated with tumor initiation and progression in various types of cancers. However, the serum level of DKK-1 in patients with papillary thyroid cancer (PTC) and its potential clinical significance is poorly understood. Enzyme-linked immunosorbent assay (ELISA) was used to evaluate the level of serum DKK-1 in patients with PTC (N = 132) and healthy controls (N = 40). The association between serum DKK-1 level and clinicopathological parameters of PTC was examined and independent prognostic markers for PTC were identified. The mean serum DKK-1 level was significantly lower in patients with PTC than healthy controls (44.64 ± 15.13 and 85.51 ± 9.94 ng/mL, respectively; P < 0.01). Following treatment, the mean serum DKK-1 level in PTC patients significantly increased (67.03 ± 17.09 ng/mL; P < 0.01). Serum DKK-1 level was associated with various PTC clinical features including tumor size (P = 0.003), lymph node metastasis (P = 0.001), and tumor-node-metastasis stage (P = 0.004). Survival analysis revealed that PTC patients who had lower serum DKK-1 levels suffered both poorer overall survival (P = 0.036) and relapse-free survival (P = 0.015). Moreover, serum DKK-1 levels were an independent risk factor for predicting the prognosis of PTC (P = 0.031). In conclusion, low DKK-1 serum levels are associated with poor prognosis in PTC patients and DKK-1 could potentially be used as a biomarker leading to earlier diagnosis of PTC.
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Biomarcadores de Tumor/genética , Carcinoma/genética , Péptidos y Proteínas de Señalización Intercelular/genética , Recurrencia Local de Neoplasia/genética , Glándula Tiroides/metabolismo , Neoplasias de la Tiroides/genética , Adolescente , Adulto , Anciano , Biomarcadores de Tumor/sangre , Carcinoma/diagnóstico , Carcinoma/mortalidad , Carcinoma/patología , Carcinoma Papilar , Estudios de Casos y Controles , Regulación hacia Abajo , Ensayo de Inmunoadsorción Enzimática , Femenino , Expresión Génica , Humanos , Péptidos y Proteínas de Señalización Intercelular/sangre , Metástasis Linfática , Masculino , Persona de Mediana Edad , Recurrencia Local de Neoplasia/diagnóstico , Recurrencia Local de Neoplasia/mortalidad , Recurrencia Local de Neoplasia/patología , Estadificación de Neoplasias , Pronóstico , Factores de Riesgo , Análisis de Supervivencia , Cáncer Papilar Tiroideo , Glándula Tiroides/patología , Neoplasias de la Tiroides/diagnóstico , Neoplasias de la Tiroides/mortalidad , Neoplasias de la Tiroides/patología , Carga TumoralRESUMEN
The nucleotide-binding site (NBS) disease-resistance genes are the largest category of plant disease-resistance gene analogs. The complete set of disease-resistant candidate genes, which encode the NBS sequence, was filtered in the genomes of two varieties of foxtail millet (Yugu1 and 'Zhang gu'). This study investigated a number of characteristics of the putative NBS genes, such as structural diversity and phylogenetic relationships. A total of 269 and 281 NBS-coding sequences were identified in Yugu1 and 'Zhang gu', respectively. When the two databases were compared, 72 genes were found to be identical and 164 genes showed more than 90% similarity. Physical positioning and gene family analysis of the NBS disease-resistance genes in the genome revealed that the number of genes on each chromosome was similar in both varieties. The eighth chromosome contained the largest number of genes and the ninth chromosome contained the lowest number of genes. Exactly 34 gene clusters containing the 161 genes were found in the Yugu1 genome, with each cluster containing 4.7 genes on average. In comparison, the 'Zhang gu' genome possessed 28 gene clusters, which had 151 genes, with an average of 5.4 genes in each cluster. The largest gene cluster, located on the eighth chromosome, contained 12 genes in the Yugu1 database, whereas it contained 16 genes in the 'Zhang gu' database. The classification results showed that the CC-NBS-LRR gene made up the largest part of each chromosome in the two databases. Two TIR-NBS genes were also found in the Yugu1 genome.
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Biología Computacional/métodos , Resistencia a la Enfermedad/genética , Enfermedades de las Plantas/genética , Setaria (Planta)/genética , Secuencia de Aminoácidos , Sitios de Unión/genética , Mapeo Cromosómico , Cromosomas de las Plantas/genética , Genes de Plantas/genética , Variación Genética , Genoma de Planta/genética , Datos de Secuencia Molecular , Familia de Multigenes , Nucleótidos/metabolismo , Filogenia , Proteínas de Plantas/clasificación , Proteínas de Plantas/genética , Homología de Secuencia de Aminoácido , Setaria (Planta)/clasificación , Especificidad de la EspecieRESUMEN
Selection pressures are the principle evolutionary forces for the genetic differentiation of populations. Recent changes in selection pressures on mitochondrial DNA and microsatellite have been described in a wide variety of organisms. The common carp (Cyprinus carpio) has experienced strong selection pressure, in particular artificial selection, during its domestication. However, the contribution and extent of artificial selection in driving genome-wide population differentiation remain unclear. We investigated the genetic differentiation of 4 domesticated strains (Xingguo red common carp, Glass red common carp, Purse red common carp, and Jian common carp, which have been generated by artificial selection since 1970s) and 2 wild populations (Shishou section in Hubei and Yangzhou section in Jiangsu of the Yangtze River) of common carp in China by sequencing the mitochondrial DNA D-loop and by genotyping 10 microsatellite loci. It was found that the domesticated strains exhibited linkage disequilibrium within the population and less genetic variability, higher inbreeding coefficients (F(IS) = 0.101 vs 0.038), and higher genetic differentiation (F(ST) = 0.087 vs 0.001) than the wild populations, which indicates strong selection pressures in the process of domestication. Of the 10 loci, 5 appeared to be under positive directional selection in the domesticated strains, and all 10 loci in wild populations were potentially under balancing selection. We conclude that strong selection pressures, artificial selection in particular, have caused genetic differentiation between populations of domesticated and wild common carp.
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Animales Domésticos/genética , Animales Salvajes/genética , Carpas/genética , Selección Genética , Animales , ADN Mitocondrial/genética , Sitios Genéticos/genética , Variación Genética , Genética de Población , Haplotipos/genética , Heterocigoto , Repeticiones de Microsatélite/genética , Datos de Secuencia Molecular , Análisis de Componente PrincipalRESUMEN
We studied whether two IGF2 transcripts in common carp are similar to those found in zebrafish. The full-length IGF2a cDNA contains a 5'-terminal untranslated region (UTR) of 105 bp, a 3'-terminal UTR of 1358 bp and an open reading frame of 612 bp, which encodes a 206-amino acid protein. A 6614-bp full-length IGF2a DNA molecule, including the 5'-flanking region, was isolated. Genomic DNA structure analysis revealed that the IGF2a gene contains four exons and three introns. Bioinformatics analysis indicated that the proteins encoded by IGF2a genes in common carp have one signal peptide and one apparent transmembrane region. Bootstrapping was performed 1000 times to obtain support values for each branch. The common carp IGF2a were clustered in one group, while the outgroup (common carp IGF1) clustered in another group. We identified two new single nucleotide polymorphisms in intron 2 of the gene. One polymorphism, A/N, can be found only in the Huanghe carp. The other polymorphism, C/N, can be found in both male Huanghe carp × female Heilongjiang carp and male Huanghe carp × female Jian carp. The second polymorphism, C/N, is primarily transferred from the male and may be related to heterosis.
Asunto(s)
Carpas/genética , Proteínas de Peces/genética , Polimorfismo de Nucleótido Simple/genética , Animales , Clonación Molecular , Biología Computacional , Exones/genética , Intrones/genética , Datos de Secuencia MolecularRESUMEN
Studies on mechanisms underlying the differentiation of dental pulp stem cells are critical for the understanding of the biology of odontogenesis and for dental tissue engineering. Here, we tested the hypothesis that stem cells from exfoliated deciduous teeth (SHED) differentiate into functional odontoblasts and endothelial cells. SHED were seeded in tooth slice/scaffolds and implanted subcutaneously into immunodeficient mice. SHED differentiated into functional odontoblasts that generated tubular dentin, as determined by tetracycline staining and confocal microscopy. These cells also differentiated into vascular endothelial cells, as determined by beta-galactosidase staining of LacZ-tagged SHED. In vitro, vascular endothelial growth factor (VEGF) induced SHED to express VEGFR2, CD31, and VE-Cadherin (markers of endothelium) and to organize into capillary-like sprouts. VEGF induced ERK and AKT phosphorylation (indicative of differentiation), while inhibiting phosphorylation of STAT3 (indicative of 'stemness'). Collectively, this work demonstrates that SHED can differentiate into angiogenic endothelial cells and odontoblasts capable of generating tubular dentin.
Asunto(s)
Células Madre Adultas/citología , Pulpa Dental/citología , Dentina/metabolismo , Endotelio Vascular/citología , Neovascularización Fisiológica/fisiología , Odontoblastos/citología , Animales , Diferenciación Celular , Células Cultivadas , Células Endoteliales/citología , Células Endoteliales/efectos de los fármacos , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/metabolismo , Proteínas de la Matriz Extracelular/biosíntesis , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Humanos , Ratones , Ratones SCID , Odontoblastos/efectos de los fármacos , Odontoblastos/metabolismo , Fosfoproteínas/biosíntesis , Fosforilación , Proteínas Proto-Oncogénicas c-akt/metabolismo , Proteínas Recombinantes/farmacología , Factor de Transcripción STAT3/metabolismo , Sialoglicoproteínas/biosíntesis , Tejido Subcutáneo , Andamios del Tejido , Diente Primario/citología , Factor A de Crecimiento Endotelial Vascular/farmacología , Receptor 1 de Factores de Crecimiento Endotelial Vascular/fisiologíaRESUMEN
AIMS: To isolate and identify endophytic nitrogen-fixing bacteria in sugarcane growing in Cuba without chemical fertilizers. METHODS AND RESULTS: Two N2-fixing isolates, 9C and T2, were obtained from surface-sterilized stems and roots, respectively, of sugarcane variety ML3-18. Both isolates showed acetylene reduction and H2 production in nitrogen-free media. Nitrogenase activity measured by H2 production was about 15 times higher for isolate 9C than for T2 or for Gluconoacetobacter diazotrophicus (PAL-5 standard strain, ATCC 49037). The nifH gene segment was amplified from both isolates using specific primers. Classification of both T2 and 9C was made on the basis of morphological, biochemical, PCR tests and 16S rDNA sequence analysis. CONCLUSIONS: Isolate 9C was identified as a Pantoea species from its 16S rDNA, but showed considerable differences in physiological properties from previously reported species of this genus. For example, 9C can be cultured over a wide range of temperature, pH and salt concentration, and showed high H2 production (up to 67.7 nmol H2 h(-1) 10(10) cell(-1)). Isolate T2 was a strain of Gluconacetobacter diazotrophicus. SIGNIFICANCE AND IMPACT OF THE STUDY: A new N2-fixing endophyte, i.e. Pantoea, able to produce H2 and to grow in a wide range of conditions, was isolated from sugarcane stem tissue and characterized. The strain with these attributes may well be valuable for agriculture.