RESUMEN

In this research, compound Maqin decoction (CMD) has been shown to positively affect in airway inflammation of asthma models. We evaluated the effects of CMD on the expression of transforming growth factor (TGF)-ß1/Smad proteins, interleukin (IL)-17, and IL-10 in lung tissue of asthmatic rats. Asthma was induced in a rat model using ovalbumin. After a 4-week treatment with CMD, rats were killed to evaluate the expression of TGF-ß1 and Smad proteins in lung tissue. IL-10 and IL-17 levels in lung tissue homogenates were determined by ELISA. The expression of TGF-ß1 and Smad3 protein increased, whereas expression of Smad7 protein decreased upon high-dose or low-dose treatment with CMD or by intervention with dexamethasone, compared to the control. There was a significant difference between treatment with a high dose CMD and the control treatment, but no significant difference was found between high-dose CMD treatment and dexamethasone intervention. The expression of TGF-ß1 and Smad7 protein increased, whereas the expression of Smad3 protein decreased in the model group compared to other groups. In the CMD high-dose group, low-dose group, and dexamethasone intervention group, the IL-17 concentrations in lung tissue homogenates were decreased, while IL-10 levels were increased. Again, there was a significant difference between CMD high-dose and control treatment, but not between CMD high-dose treatment and dexamethasone intervention. Thus, positive effects of CMD against asthmatic airway remodeling may be due to its regulatory effect on TGF-ß1, Smad3, and Smad7 protein levels and on cytokines such as IL-10 and IL-17.

Asunto(s)

Antiasmáticos/farmacología , Asma/tratamiento farmacológico , Pulmón/efectos de los fármacos , Extractos Vegetales/farmacología , Factor de Crecimiento Transformador beta1/inmunología , Remodelación de las Vías Aéreas (Respiratorias)/efectos de los fármacos , Animales , Antiasmáticos/aislamiento & purificación , Asma/inducido químicamente , Asma/inmunología , Asma/patología , Berberidaceae/química , Dexametasona/farmacología , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Elaeagnaceae/química , Ephedra/química , Regulación de la Expresión Génica , Interleucina-10/genética , Interleucina-10/inmunología , Interleucina-17/genética , Interleucina-17/inmunología , Pulmón/inmunología , Pulmón/patología , Masculino , Ovalbúmina , Extractos Vegetales/química , Ratas , Ratas Sprague-Dawley , Scutellaria baicalensis/química , Transducción de Señal , Proteína smad3/genética , Proteína smad3/inmunología , Proteína smad7/genética , Proteína smad7/inmunología , Factor de Crecimiento Transformador beta1/genética , Xanthium/química

RESUMEN

MicroRNAs (miRNAs) are key regulators of gene expression and play an important role in the development and progression of various diseases including esophageal squamous cell carcinoma (ESCC). In this study, we determined whether a polymorphism at the miR-214 binding site in the 3'-untranslated region (3'-UTR) of the methylenetetrahydrofolate reductase gene (MTHFR) is associated with susceptibility to ESCC. A total of 448 ESCC cases and 460 gender- and age-matched subjects were recruited for the study. The genotypes of the rs114673809 single nucleotide polymorphism (SNP) were determined by polymerase chain reaction sequencing. Associations between genotypes of MTHFR rs114673809 and ESCC risk were determined using logistic regression analyses. In the recessive model, when the MTHFR rs114673809 GG homozygote genotype was used as the reference group, the GA genotype was not associated with the risk of ESCC (GA vs GG: OR = 1.261, 95%CI = 0.960-1.657, P = 0.110), but the AA genotype was associated with increased risk of ECSS (AA vs GG: OR = 1.752, 95%CI = 1.076-2.853, P = 0.027). Additionally, the rs114673809 A allele carriers also showed a 1.286-fold increased ESCC risk compared with those carrying the rs114673809 G allele genotype. Furthermore, we observed a significant increase in plasma homocysteine levels in ESCC cases carrying the AA genotype relative to ESCC cases carrying the GG genotype. Our data demonstrate that a polymorphism at the miR-214 binding site in the 3'-UTR of MTHFR is an ESCC susceptibility SNP in the Chinese population.

Asunto(s)

Carcinoma de Células Escamosas/genética , Neoplasias Esofágicas/genética , Metilenotetrahidrofolato Reductasa (NADPH2)/genética , MicroARNs/genética , Regiones no Traducidas 3' , Adulto , Anciano , Alelos , Pueblo Asiatico/genética , Sitios de Unión/genética , Carcinoma de Células Escamosas/enzimología , Carcinoma de Células Escamosas/metabolismo , Estudios de Casos y Controles , Neoplasias Esofágicas/enzimología , Neoplasias Esofágicas/metabolismo , Carcinoma de Células Escamosas de Esófago , Femenino , Predisposición Genética a la Enfermedad , Genotipo , Humanos , Masculino , Metilenotetrahidrofolato Reductasa (NADPH2)/metabolismo , MicroARNs/metabolismo , Persona de Mediana Edad , Polimorfismo de Nucleótido Simple

RESUMEN

Habitual abortion is associated with the altered expression of multiple genes. This study was carried out to investigate the relationship between expression of Toll-like receptor 4 (TLR4) and monocyte chemotactic protein 2 (MCP2 or CCL8) and habitual abortion. This was done by detecting and comparing their relative expression in peripheral blood and placental villi of patients and healthy fertile women. Based on our previous research, 85 subjects with habitual abortion (study group) and 40 healthy fertile women (control group), who were admitted to our hospital between June 2013 and December 2014, were enrolled in this study. After these subjects signed written informed consent, peripheral blood samples and villous tissues were collected, from which the total RNA was extracted. The expression of TLR4 and MCP2 was detected with quantitative reverse transcription-polymerase chain reaction, using GAPDH as a reference control. The expression of TLR4 and MCP2 in the peripheral blood and villous tissues of the study group was significantly higher than that of the control group (P < 0.05). A positive correlation was also observed between the changes in expression levels of TLR4 and MCP2. In conclusion, TLR4 and MCP2 expression correlated with the occurrence of habitual abortion. Detecting expression changes in TLR4 and MCP2 in the peripheral blood is a feasible method for predicting the occurrence of abortion in women of child-bearing age.

Asunto(s)

Aborto Habitual/genética , Quimiocina CCL8/biosíntesis , Receptor Toll-Like 4/biosíntesis , Aborto Habitual/patología , Adulto , Quimiocina CCL8/genética , Vellosidades Coriónicas/metabolismo , Femenino , Regulación de la Expresión Génica , Humanos , Embarazo , Receptor Toll-Like 4/genética

RESUMEN

Despite more than a century of intensive study, the mechanisms of successful pregnancy remain unclear. Recent research suggests that NF-κB (nuclear factor kappa B) plays an important role in embryo implantation. In the current study, we aimed to identify SNPs that contribute to genetic susceptibility for recurrent implantation failure (RIF). Thus, we examined the potential associations between RIF and ten SNPs (rs28362491, rs3774932, rs1598856, rs230528, rs230521, rs3774956, rs4648055, rs3774964, rs4648068, and rs3774968) of the NF-κB gene. Participants included 209 patients with RIF and 395 controls. Our results revealed that there were statistically significant differences observed in the allelic and genotypic frequencies of the rs28362491 promoter in the NF-κB gene. The frequency of the del/ del genotype was significantly higher in RIF patients than in healthy controls (P = 0.004). Compared with healthy controls, the RIF patients carried a higher frequency of the rs28362491 del allele (P = 0.010). Furthermore, strong linkage disequilibrium was observed in the three identified haplotype blocks (D' > 0.9). Particularly, in block 1 (rs230528-rs230521), the A-C haplotype occurred significantly more frequently (P = 0.029) in subjects with RIF (P = 0.0003). In contrast, the A-G haplotype occurred significantly less frequently (P = 0.008) in RIF subjects. These findings support an important role for G-712A polymorphisms of NF-κB in RIF, and may guide future studies that aim to characterize genetic risk factors for RIF.

Asunto(s)

Implantación del Embrión/genética , FN-kappa B/genética , Polimorfismo de Nucleótido Simple , Adulto , Estudios de Casos y Controles , Transferencia de Embrión/efectos adversos , Femenino , Haplotipos , Humanos , Desequilibrio de Ligamiento , Embarazo

RESUMEN

The hypothalamus is an important component of the nervous system, and neuropeptide Y (NPY), proopiomelanocortin (POMC), and neuromedin U (NMU) are key players in physiological regulation. Puerarin is important for nerve regulation. We investigated the effect of puerarin on the expression of NMU, NPY, and POMC genes in the hypothalamus. The results showed that the puerarin low-dose group and the other groups were significantly different (P < 0.05). However, there was no significant difference in NMU, POMC, and NPY among the groups.

Asunto(s)

Hipotálamo/metabolismo , Isoflavonas/farmacología , Neuropéptido Y/genética , Neuropéptidos/genética , Proopiomelanocortina/genética , Animales , Regulación de la Expresión Génica , Hipotálamo/efectos de los fármacos , Neuropéptido Y/efectos de los fármacos , Neuropéptidos/efectos de los fármacos , Proopiomelanocortina/efectos de los fármacos , Ratas

RESUMEN

Changes in the expression of the anti-apoptotic protein cFLIP (cellular FLICE inhibitory protein) were examined in the caprine corpus luteum (CL), during development and subsequent maintenance. Corpora lutea at four different stages were collected from Shiba goats, to measure the expression of cFLIP mRNA, protein and immunolocalization. Expression of short form cFLIP (cFLIPS) mRNA was highest at the early CL stage, and decreased during late and regressed stages (P < 0.05). In contrast, long form cFLIP (cFLIPL) mRNA expression was high during early, mid and late stages, and only decreased at the regressed stage (P< 0.01). Protein expression of cFLIPS was highest at the late CL stage, and decreased at the regressed stage (P < 0.01). Protein expression of cFLIPL was highest at the early and mid CL stages, and decreased by the late and regressed stages (P < 0.01). Further expression of cFLIPL was higher at the early CL stage than at the mid stage (P < 0.01). cFLIP protein expression was detectable by immunostaining in the early, mid and late CL stages, but not at the regressed CL stage. These results are consistent with the hypothesis that cFLIP acts as a survival factor in the maintenance of CL function in goats.

Asunto(s)

Proteína Reguladora de Apoptosis Similar a CASP8 y FADD/genética , Proteína Reguladora de Apoptosis Similar a CASP8 y FADD/metabolismo , Cuerpo Lúteo/metabolismo , Ciclo Estral/genética , Ciclo Estral/metabolismo , Cabras/fisiología , Animales , Femenino , Cabras/genética , ARN Mensajero/genética

RESUMEN

Porcine circovirus type 2 (PCV2) is considered to be the main pathogen in PC-associated diseases, and significantly affects the global pig-producing industry. PCV2 continuously evolves by point mutations and genome recombinations. In the present study, we aimed to further identify recombinant PCV2 strains. We used polymerase chain reaction to detect PCV2 in the carcasses of pigs with suspected infections from different regions of Guangdong Province in China. DNA was extracted from samples with confirmed infection and full- genome amplification, sequencing, phylogenetic tree construction, gene recombination detection, and sequence alignment were performed in gene recombination analysis. Our results show that recombination occurred between the strains SHC (DQ104421) and ZhuJi2003 (AY579893). The recombination resulted in three recombinants: GD003 (KM503044), GD005 (KM487708), and GD008 (KM487709). Further analyses revealed that these novel recombinants appeared to result from recombination between the PCV2a and PCV2b strains, with crossover regions located in ORF2. This study was a comprehensive analysis that used several different methods, which demonstrated that a cluster of PCV2 strains resulted from the same type of inter-genotypic recombination pattern, with a breakpoint in the structural protein coding region. The results of our study provide both information on the recombination mechanism and disease pathogenesis and useful data for the prevention of PCV2 in the swine industry.

Asunto(s)

Infecciones por Circoviridae/virología , Circovirus/genética , ADN Viral/genética , Virus Reordenados/genética , Recombinación Genética , Animales , Secuencia de Bases , Línea Celular , Infecciones por Circoviridae/patología , Circovirus/clasificación , Circovirus/patogenicidad , Células Epiteliales/patología , Células Epiteliales/virología , Ganglios Linfáticos/patología , Ganglios Linfáticos/virología , Datos de Secuencia Molecular , Filogenia , Virus Reordenados/patogenicidad , Alineación de Secuencia , Bazo/patología , Bazo/virología , Porcinos

RESUMEN

Moso bamboo is a large woody bamboo with the highest ecological, economic, and cultural value among all bamboos in Asia. However, environmental stress influences its growth and development and limits its geographic distribution. Therefore, improving its resistance to environmental stress is extremely important. Dehydration responsive element binding (DREB) transcription factors perform an important role in the regulation of stress-related genes, enhancing the resistance of plants to abiotic stress. In the current study, two novel DREB genes, PeDREB2A and PeDREB1A (Gene ID No. PH01000046G1730 and PH01000668G0350), were isolated from moso bamboo and the sequences were identified and characterized (coding sequence lengths were 795 and 825 bp, respectively). The PeDREB2A and PeDREB1A proteins were estimated to have typical AP2/ERF domains, molecular weights of 28.96 and 28.84 kDa, and isoelectric points of 9.47 and 5.34, respectively. RT-PCR analysis revealed that PeDREB2A and PeDREB1A were tissue-specific genes, expressed in leaves, young stems, and roots, with similar expression levels in leaves and young stems. qRT-PCR analysis of leaves demonstrated that PeDREB2A transcription levels rapidly accumulate following exposure to drought and salt stress, peaking at 12 and 0.5 h, respectively, but only low expression levels were observed under cold stress. PeDREB1A exhibited a strong response to cold stress, reaching a peak in expression 3 h after exposure, but demonstrated only a slight response to drought and salt stress. In roots, PeDREB2A was down-regulated, and PeDREB1A was initially upregulated but then declined, under stress conditions. Two plant expression vectors, pCAMBIA2300- CaMV35S-PeDREB2A and pCAMBIA2300-CaMV35S-PeDREB1A were also successfully constructed.

Asunto(s)

Genes de Plantas , Proteínas de Plantas/genética , Poaceae/genética , Poaceae/parasitología , Estrés Fisiológico/genética , Secuencia de Aminoácidos , Secuencia de Bases , Clonación Molecular , Electroforesis en Gel de Agar , Exones/genética , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Vectores Genéticos/metabolismo , Intrones/genética , Datos de Secuencia Molecular , Especificidad de Órganos/genética , Filogenia , Hojas de la Planta/genética , Hojas de la Planta/metabolismo , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Raíces de Plantas/genética , Reacción en Cadena de la Polimerasa , Homología de Secuencia de Aminoácido

RESUMEN

The swimming crab, Portunus trituberculatus, is widely distributed throughout the coastal waters of Asian-Pacific nations and is an important economic species in this region. The aquaculture of swimming crabs has been plagued by problems associated with low growth rates, poor flesh quality, and weak disease resistance. To overcome these problems, selective breeding programs have been suggested as a means of genetically improving these traits in stock populations. In this study, we evaluated the genetic differentiation of 3 different geographical populations (Zhoushan: S; Laizhou Bay: L; and Haizhou Bay: H) using 8 polymorphic microsatellite loci. Nine strains of first filial generation were obtained, with 3 geographically populations as parental stock. We assessed the growth and survival rates of the F1 generation to identify new strains or breeds showing improvements in these economically important traits. Our results indicated that pairwise FST among populations was significantly higher than 0 (P = 0.0000) for every population pair, ranging from 0.0810 to 0.1083 for 3 different geographical populations. We observed significant heterosis for the growth and viability (survival) traits, although some strains (crossbred combinations) showed evidence of hybrid weakness in some growth measurements. One particular strain ("SL") outperformed other combinations, displaying the greatest extent of heterosis over the growth and viability (survival) traits. These results indicate that hybridization may be used to increase the performance of P. trituberculatus in aquaculture.

Asunto(s)

Braquiuros/genética , Variación Genética , Animales , Braquiuros/crecimiento & desarrollo , Genética de Población , Fenotipo

RESUMEN

This study aimed to compare the effects of lipopolysaccharide (LPS) on stearoyl-coenzyme A desaturase (SCD) gene expression in mouse primary hepatic cells. To obtain sufficient total RNA, primary hepatic cells were plated on 6-cm diameter-type collagen 1-coated dishes (1 x 106 cells per dish). The test was divided into 6 groups with 6 replications per group. The 6 groups were treated with the following volumes of LPS (0.1 mg/mL): 0, 1, 1.5, 2, 4, and 8 µL. The cells were cultured for 24 h, and the total RNA was extracted from samples. Reverse transcription polymerase chain reaction was used to analyze SCD mRNA levels. With increasing LPS amounts, the SCD mRNA expression first decreased and then increased slightly; the expression was the lowest in the 2-µL LPS condition. The SCD mRNA levels from the 4- and 8-µL LPS conditions were slightly higher than that from the 2-µL LPS condition, but the difference was not significant (P > 0.05). The SCD mRNA level from the 2-µL LPS condition was obviously lower than that from the 0-, 1-, and 1.5-µL LPS condition, and the differences were significant (P < 0.05), and the SCD mRNA levels from the 0-, 1-, and 1.5-µL LPS conditions were not significantly different (P > 0.05). The SCD mRNA levels from the 4- and 8-µL LPS conditions were obviously lower than those from the 0- and 1-µL LPS conditions, and the differences were significant (P < 0.05).

Asunto(s)

Hepatocitos/efectos de los fármacos , ARN Mensajero/biosíntesis , Estearoil-CoA Desaturasa/biosíntesis , Animales , Bovinos , Regulación de la Expresión Génica/efectos de los fármacos , Lipopolisacáridos/administración & dosificación , Ratones , Estearoil-CoA Desaturasa/genética

RESUMEN

The liver is a unique organ that is endowed with a plethora of specialized functions. Most of its functional traits are controlled by hepatocytes. Primary hepatocytes have been used widely in in vitro models to understand the biological processes occurring in the liver. There are a number of methods used to separate hepatocytes, but the cell activity and purity are much lower in this condition. On the basis of previous research, in this study, the two-step collagenase perfusion technique was used for isolating hepatocytes. The key proteins of hepatocytes, cytokeratin-18 (CK-18) and albumin (ALB), were used to identify cells, and their contents were evaluated by immunohistochemistry and Western blotting. The results showed that the isolated hepatocytes comprised more than 96% of the corresponding protein volume stability. Therefore, this method was demonstrated to be reliable for identifying hepatocytes.

Asunto(s)

Separación Celular , Hepatocitos/citología , Hepatocitos/metabolismo , Animales , Biomarcadores , Bovinos , Separación Celular/métodos , Cultivo Primario de Células

RESUMEN

It has been shown that cytokines can act as molecular adjuvant to enhance the immune response induced by DNA vaccines, but it is unknown whether interleukin 33 (IL-33) can enhance the immunocontraceptive effect induced by DNA vaccines. In the present study, we explored the effects of murine IL-33 on infertility induced by Lagurus lagurus zona pellucida 3 (Lzp3) contraceptive DNA vaccine administered by the mucosal route. Plasmid pcD-Lzp3 and plasmid pcD-mIL-33 were encapsulated with chitosan to generate the nanoparticle chi-(pcD-Lzp3+pcD-mIL-33) as the DNA vaccine. Sixty female ICR mice, divided into 5 groups (n=12/group), were intranasally immunized on days 0, 14, 28, and 42. After intranasal immunization, the anti-LZP3-specific IgG in serum and IgA in vaginal secretions and feces were determined by ELISA. The results showed that chi-(pcD-Lzp3+pcD-mIL-33) co-immunization induced the highest levels of serum IgG, secreted mucosal IgA, and T cell proliferation. Importantly, mice co-immunized with chi-(pcD-Lzp3+pcD-mIL-33) had the lowest birth rate and mean litter size, which correlated with high levels of antibodies. Ovaries from infertile female mice co-immunized with chi-(pcD-Lzp3+pcD-mIL-33) showed abnormal development of ovarian follicles, indicated by atretic follicles and loss of oocytes. Our results demonstrated that intranasal delivery of the molecular adjuvant mIL-33 with chi-pcD-Lzp3 significantly increased infertility by enhancing both systemic and mucosal immune responses. Therefore, chi-(pcD-Lzp3+pcD-mIL-33) co-immunization could be a strategy for controlling the population of wild animal pests.

RESUMEN

It has been shown that cytokines can act as molecular adjuvant to enhance the immune response induced by DNA vaccines, but it is unknown whether interleukin 33 (IL-33) can enhance the immunocontraceptive effect induced by DNA vaccines. In the present study, we explored the effects of murine IL-33 on infertility induced by Lagurus lagurus zona pellucida 3 (Lzp3) contraceptive DNA vaccine administered by the mucosal route. Plasmid pcD-Lzp3 and plasmid pcD-mIL-33 were encapsulated with chitosan to generate the nanoparticle chi-(pcD-Lzp3+pcD-mIL-33) as the DNA vaccine. Sixty female ICR mice, divided into 5 groups (n=12/group), were intranasally immunized on days 0, 14, 28, and 42. After intranasal immunization, the anti-LZP3-specific IgG in serum and IgA in vaginal secretions and feces were determined by ELISA. The results showed that chi-(pcD-Lzp3+pcD-mIL-33) co-immunization induced the highest levels of serum IgG, secreted mucosal IgA, and T cell proliferation. Importantly, mice co-immunized with chi-(pcD-Lzp3+pcD-mIL-33) had the lowest birth rate and mean litter size, which correlated with high levels of antibodies. Ovaries from infertile female mice co-immunized with chi-(pcD-Lzp3+pcD-mIL-33) showed abnormal development of ovarian follicles, indicated by atretic follicles and loss of oocytes. Our results demonstrated that intranasal delivery of the molecular adjuvant mIL-33 with chi-pcD-Lzp3 significantly increased infertility by enhancing both systemic and mucosal immune responses. Therefore, chi-(pcD-Lzp3+pcD-mIL-33) co-immunization could be a strategy for controlling the population of wild animal pests.