RESUMEN
Studying thermotolerance is important for the prevention of thermostress in chickens. This study aimed to analyze the effect of mutations in the heat shock protein 70 (HSP70) gene on chicken thermotolerance. The C.-69A>G SNP in the 5'-flanking region of the HSP70 gene was genotyped in Lingshan and White Recessive Rock (WRR) chickens. Association of this SNP with thermotolerance traits revealed it to be significantly associated with CD4+/CD8+, and potentially associated with heterophil-to-lymphocyte ratio in WRR chickens exposed to thermoneutral temperature (15°C). Online prediction detected a putative myeloid zinc finger protein 1 binding factor in the C.-69A>G mutation. Under acute thermostress, mRNA levels of HSP70 in individuals with different C.-69A>G genotypes varied in the heart, leg muscle, and liver tissues. The HSP70 protein was expressed at higher levels in individuals with the GG genotype than in those with the AA genotype. In heart and liver, protein expression of HSP70 in individuals with the GG genotype was significantly higher than in those with the AA genotype. In leg muscle, protein expression was higher in birds with the GG genotype than in those with the AA and AG genotypes. Luciferase activity of the GG genotype was significantly higher than that of the AA genotype, suggesting that the C.-69A>G SNP regulates HSP70 gene expression. These results indicate that the C.-69A>G SNP in the 5'-flanking region of the HSP70 gene might affect chicken thermotolerance and that the GG genotype might be advantageous for the prevention of thermostress.
Asunto(s)
Pollos/fisiología , Proteínas HSP70 de Choque Térmico/genética , Termotolerancia/genética , Región de Flanqueo 5' , Animales , Pollos/genética , Femenino , Genotipo , Mutación , Polimorfismo de Nucleótido Simple , ARN Mensajero/metabolismoRESUMEN
Numerous studies have investigated the relationship between the interleukin-1ß gene (IL1B) -511C/T polymorphism and ischemic stroke (IS) risk. However, the results are inconsistent. We performed this meta-analysis of all available case-control studies that evaluated the relationship between the IL1B -511C/T polymorphism and IS. Studies were retrieved from the PubMed and Embase databases. Statistical analyses were conducted using the STATA 11.0 software. Odds ratios (ORs) with 95% confidence intervals (95%CIs) were applied to determine the strength of association. Nine studies comprising a total of 2072 patients and 2173 controls were included. No significant variation in IS risk was detected in any of the genetic models (CC vs TT: OR = 0.78, 95%CI = 0.48-1.27; CT vs TT: OR = 0.83, 95%CI = 0.62-1.10; dominant model: OR = 0.79, 95%CI = 0.55-1.15; recessive model: OR = 0.90, 95%CI = 0.66-1.24). Taking into account the effects of race, further subgroup analyses were performed and our results showed no association between the IL1B gene -511C/T polymorphism and IS in either Asians or Caucasians. No publication bias was found in our study. In conclusion, the IL1B gene -511C/T polymorphism might not be associated with IS risk.
Asunto(s)
Isquemia Encefálica/genética , Estudios de Asociación Genética , Predisposición Genética a la Enfermedad , Interleucina-1beta/genética , Accidente Cerebrovascular/genética , Pueblo Asiatico , Isquemia Encefálica/patología , Femenino , Humanos , Masculino , Polimorfismo de Nucleótido Simple , Factores de Riesgo , Accidente Cerebrovascular/patologíaRESUMEN
Dendrobium officinale is one of the most well-known traditional Chinese medicines, and polysaccharide is its main active ingredient. Many studies have investigated the synthesis and accumulation mechanisms of polysaccharide, but until recently, little was known about the molecular mechanism of how polysaccharide is synthesized because no related genes have been cloned. In this study, we cloned an alkaline/neutral invertase gene from D. officinale (DoNI) by the rapid amplification of cDNA ends (RACE) method. DoNI was 2231 bp long and contained an open reading frame that predicted a 62.8-kDa polypeptide with 554-amino acid residues. An alkaline/neutral invertase conserved domain was predicted from this deduced amino acid sequence, and DoNI had a similar deduced amino acid sequence to Setaria italica and Oryza brachyantha. We also found that DoNI expression in different tissues was closely related to DoNI activity, and more importantly, polysaccharide level. Our results indicate that DoNI is associated with polysaccharide accumulation in D. officinale.
Asunto(s)
Dendrobium/genética , Genes de Plantas , Proteínas de Plantas/genética , Polisacáridos/metabolismo , beta-Fructofuranosidasa/genética , Secuencia Conservada , Dendrobium/enzimología , Sistemas de Lectura Abierta , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Polisacáridos/genética , Dominios Proteicos , beta-Fructofuranosidasa/química , beta-Fructofuranosidasa/metabolismoRESUMEN
We investigated azoospermia region microdeletions in male infertility patients with Klinefelter syndrome (KFS), as well as the association between azoospermia symptoms in patients with KFS and Y chromosome microdeletion polymorphisms. A total of 111 cases with male infertility confirmed to have KFS (47, XXY) and 94 fertile men were included in this study. Peripheral blood was drawn and DNA was extracted from these samples. Multiplex polymerase chain reaction was performed to screen the partial deletions of 25 sequence-tagged sites on the Y chromosome. In 111 cases with KFS, 1 case contained the AZFb+d+c deletion. The Gr/Gr deletion was identified in 12 KFS cases and 5 control cases. In addition, the b2/b3 deletion was identified in 13 KFS cases and 6 control cases. There were no significant differences in phenotype and genotype of the 2 partial AZFc deletions between patients and controls (P > 0.05). Our results suggest that patients with KFS may also have Y chromosome microdeletions to varying degrees and that the gr/gr deletion and b2/b3 deletion may not play a role in the susceptible genetic background of azoospermia in patients with KFS in the Sichuan population.
Asunto(s)
Síndrome de Klinefelter/genética , Proteínas de Plasma Seminal/genética , Azoospermia/genética , Deleción Cromosómica , Cromosomas Humanos Y/genética , Eliminación de Gen , Sitios Genéticos/genética , Humanos , Infertilidad Masculina/genética , Masculino , Fenotipo , Lugares Marcados de Secuencia , Aberraciones Cromosómicas Sexuales , Trastornos de los Cromosomas Sexuales del Desarrollo Sexual/genéticaRESUMEN
The aim of this study was to observe the influence of gap junction (GJ) functional changes on the hepatotoxicity of TNF-α. Three different methods were employed to study functional effects of the GJ inhibition: 1) pretreatment with a GJ inhibitor; 2) inoculation of cells at high and low densities; and 3) inhibition of the expression of connexin 32 (Cx32) by small inhibitory RNA transfection. We then observed the influence of these treatments on hepatotoxicity following treatment with different concentrations of TNF-α for various duration. The hepatotoxicity of TNF-α was observed to occur in a dose- and time-dependent manner; after pretreatment inhibition, the hepatotoxicity of TNF-α was significantly reduced (P < 0.01). The hepatotoxicity of TNF-α was also found to be remarkably lower in cells that had been inoculated at low density (as measured by the amount of GJ formation among cells) than in those inoculated at density (P < 0.01). In addition, following Cx32 inhibition, the hepatotoxicity of TNF-α was significantly decreased (P < 0.01) as well. Together, these results suggest that inhibition of GJ function or of its component Cx32 significantly decreases the hepatotoxicity of TNF-α, and that the expression of Cx32 plays an important role in the hepatotoxicity of TNF-α.
Asunto(s)
Conexinas/metabolismo , Uniones Comunicantes/metabolismo , Hepatocitos/metabolismo , Animales , Línea Celular , Conexinas/genética , Hepatocitos/efectos de los fármacos , Ratas , Factor de Necrosis Tumoral alfa/toxicidad , Proteína beta1 de Unión ComunicanteRESUMEN
Previous studies have shown that genetic polymorphisms in exon7 of the NSUN7 gene can be used as an infertility marker in Iranian men with asthenospermia. However, there have been no equivalent studies in China. In the present study, we investigated the possible association between the genetic polymorphisms in exon7 of NSUN7 and asthenospermia in a Chinese Han population. We recruited 240 asthenospermic men as a patient group and 256 normospermic men as a control group, and analyzed the semen parameters on the basis of World Health Organization (WHO) guidelines. The genetic polymorphisms in exon7 of NSUN7 were detected by DNA sequence analysis. The results were analyzed statistically and a P value < 0.05 was considered significant. There were two genetic polymorphisms, c.906C>T and c.922T>G, in exon7 of NSUN7. We found relatively similar genotypes and allele frequencies between the two groups (P = 0.928, P = 0.928, respectively). The combined genotypes of the two polymorphisms did not identify a haplotype associated with asthenospermia (P = 0.824, P = 0.824, respectively). Our findings revealed that genetic polymorphisms in exon7 of the NSUN7 gene are not associated with asthenospermia in Chinese Han men.
Asunto(s)
Pueblo Asiatico/genética , Astenozoospermia/genética , Exones , Polimorfismo de Nucleótido Simple , Adulto , Alelos , Estudios de Casos y Controles , China , Frecuencia de los Genes , Genotipo , Haplotipos , Humanos , Masculino , Persona de Mediana Edad , Análisis de Secuencia de ADN , Adulto JovenRESUMEN
The regulatory region of the ribulose-1,5-bisphosphate carboxylase small subunit gene SRS4 from soybean (Glycine max) was cloned using TAIL-PCR and general PCR, and named the rbcS promoter. The promoter was fused with the GUS gene and introduced into Nicotiana tabacum via Agrobacterium-mediated leaf disk transformation. In 4-week-old transgenic tobacco plants, the highest GUS expression levels were observed in the leaves, GUS activity was 7.13- and 7.40-fold higher in leaves than in stems and roots, respectively. Moreover, GUS activity was stimulated by light. In conclusion, spatial and light regulation of the soybean rbcS promoter was observed in N. tabacum, thus illustrating a leaf-specific and light-induced promoter.
Asunto(s)
Glycine max/genética , Nicotiana/genética , Ribulosa-Bifosfato Carboxilasa/genética , Secuencia de Aminoácidos , Secuencia de Bases , Clonación Molecular , Regulación de la Expresión Génica de las Plantas , Luz , Datos de Secuencia Molecular , Hojas de la Planta/genética , Plantas Modificadas Genéticamente/genética , Reacción en Cadena de la Polimerasa/métodos , Regiones Promotoras Genéticas , Glycine max/enzimología , Nicotiana/enzimologíaRESUMEN
Sauvagesia rhodoleuca (Ochnaceae) is an endangered plant that is endemic to southern China. The levels of genetic variation and patterns of population structure in S. rhodoleuca were investigated using inter-simple sequence repeat markers. Eleven primers were used to amplify DNA samples from 117 individuals, and a total of 92 loci were detected. Our results indicated that genetic diversity was quite low both at the species level (percentage of polymorphic bands (PPB) = 41.30%, Nei's gene diversity (h) = 0.1331, and Shannon information index (I) = 0.2028) and the population level (PPB = 16.30-28.26%, h = 0.0496-0.1012, and I = 0.0756-0.1508). A high level of genetic differentiation among populations was detected based on Nei's genetic diversity analysis (0.4344) and analysis of molecular variance (47.03%). The low genetic diversity within population and high population differentiation of S. rhodoleuca were assumed to result largely from limited gene flow, genetic drift, inbreeding, and clonal growth. Conservation strategies for this endangered species are proposed based on the genetic data.
Asunto(s)
Variación Genética , Genética de Población , Repeticiones de Microsatélite , Ochnaceae/genética , China , Especies en Peligro de Extinción , Evolución Molecular , Geografía , Especies IntroducidasRESUMEN
MicroRNAs (miRNAs) are small non-coding RNAs that regulate the translation of targeted mRNAs. An increasing amount of evidence indicates that miRNAs play important role in cancer pathogenesis, apoptosis, proliferation, and differentiation as oncogenes or tumor suppressors. Recently, miRNA-199a has been shown to be involved in many human cancers, although the role of miRNA-199a-3p in gastric cancer has not yet been evaluated. In the present study, the expression of miRNA-199a-3p was found to be significantly downregulated in human gastric cancer tissues and cells. miRNA-199a-3p induced anti-proliferation effects on human gastric cancer cells. Furthermore, using quantitative RT-PCR (real-time polymerase chain reaction) and luciferase reporter assays, mTOR was identified as a direct target gene of miRNA-199a-3p that is downregulated by it. In conclusion, our findings suggest that miRNA-199a-3p is associated with human gastric cancer through its ability to decrease cancer cell proliferation and target the mTOR signaling pathway, and, therefore, may provide a novel therapeutic target for the treatment of human gastric cancer.
Asunto(s)
Proliferación Celular , MicroARNs/biosíntesis , Neoplasias Gástricas/genética , Serina-Treonina Quinasas TOR/genética , Apoptosis/genética , Línea Celular Tumoral , Regulación hacia Abajo , Regulación Neoplásica de la Expresión Génica , Humanos , MicroARNs/genética , Neoplasias Gástricas/patología , Serina-Treonina Quinasas TOR/metabolismoRESUMEN
DNA methylation plays a central role in gene expression. In this study, we detected the promoter methylation pattern of the chicken heat shock protein 70 (HSP70) gene and its association with messenger RNA (mRNA) expression before and after heat shock. The results showed that mRNA expression increased in response to heat stress and peaked at 3 h before dropping. Hypomethylation of the HSP70 promoter occurred in all of the groups studied, but the difference between groups within tissue type was not significant. The DNA methylation level of the control and the 6-h treatment groups was slightly higher than that of the 3-h treatment group in brain tissue and leg muscle. Correlation analysis between mRNA expression and DNA methylation of HSP70 showed that DNA methylation was negatively associated with mRNA expression in leg muscle (P = 0.0124), indicating that DNA methylation may be negatively associated with the expression of HSP70, although the difference was not significant. We concluded that the expression of HSP70 is heat inducible and tissue dependent and that heat induction may correlate with DNA methylation pattern in the HSP70 promoter, whereas tissue dependence is unrelated to DNA methylation pattern.