RESUMO
PURPOSE: To investigate the role of PRDX2 in esophageal carcinoma (ESCA). METHODS: The expression of PRDX2 was detected in ESCA tissues. And PRDX2 expression in two ESCA cell lines was knocked down. Cell proliferation, metastasis and invasion were detected in these cells. RESULTS: Here, we found that PRDX2 expression was significantly increased in ESCA tissues and was associated with a poor prognosis in ESCA patients. In addition, PRDX2 expression was significantly associated with pathological grading, infiltration degree and 5-year survival time in ESCA patients. Next, we knocked down PRDX2 expression by PRDX2-shRNA transfection in two ESCA cell lines, Eca-109 and TE-1. Proliferation analysis indicated that in vitro PRDX2 knockdown decreased growth and clone formation of ESCA cells. Scratch and transwell assays indicated that cell migration and invasion were significantly inhibited by PRDX2 knockdown. In addition, PRDX2 knockdown inhibited cell cycle of ESCA cells and down-regulated Cyclin D1-CDK4/6. Moreover, PRDX2 knockdown regulated proteins involved in mitochondrial-dependent apoptosis, including increased Bax and Caspase9/3 and decreased Bcl2. Mechanism investigation indicated that PRDX2 knockdown led to inactivation of Wnt/ß-catenin and AKT pathways. CONCLUSIONS: Our data suggest that PRDX2 may function as an oncogene in the development of ESCA via regulating Wnt/ß-catenin and AKT pathways. Our study fills a gap in the understanding of the role of PRDX2 in ESCA and provides a potential target for ESCA treatment.
Assuntos
Neoplasias Esofágicas/etiologia , Carcinoma de Células Escamosas do Esôfago/etiologia , Peroxirredoxinas/fisiologia , Proteínas Proto-Oncogênicas c-akt/fisiologia , Via de Sinalização Wnt/fisiologia , Apoptose , Pontos de Checagem do Ciclo Celular , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Neoplasias Esofágicas/patologia , Carcinoma de Células Escamosas do Esôfago/patologia , Humanos , Peroxirredoxinas/análiseRESUMO
A dose-response experiment with four dietary copper concentrations (4.17, 8.17, 12.17 and 16.17 mg/kg) was conducted to estimate the growth performance, slaughter performance, nutrient content of fecal and liver copper concentrations of growing Goslings from 28 to 70 d of age. Two hundred healthy male Yangzhou geese with similar body weight were randomized to four groups with five replicates per treatment and ten geese per replicate. Average daily feed intake, average daily gain and feed conversion ratio of geese for each pen were measured from 28 to 70 d of age. At 70 d of age, two geese were selected randomly from each pen and slaughtered to evaluate carcass quality. Metabolism experiment was conducted with five male geese from each group (one goose per pen) which body weight was close to the mean weight of the group from 64 to 70 d of age. Significant effects of dietary copper was found on body weight, feed conversion ratio, carcass yield, fecal copper concentrations and liver copper concentrations. Body weight, feed conversion ratio and carcass yield showed significant quadratic response to increase dietary copper concentration, while fecal copper concentration and liver copper concentration showed a significant linear response. The result showed that dietary Cu addition can improve growth by increasing the use of the feeding stuff and improving carcass yield in growing Goslings. Furthermore, taking into consideration, the optimal level of Gosling dietary copper was between 8.77 and 11.6 mg/kg from 28 to 70 days of age.(AU)
Assuntos
Animais , Recém-Nascido , Cobre/análise , Abate de Animais , Gansos/anormalidades , Gansos/fisiologia , Fezes/químicaRESUMO
A dose-response experiment with four dietary copper concentrations (4.17, 8.17, 12.17 and 16.17 mg/kg) was conducted to estimate the growth performance, slaughter performance, nutrient content of fecal and liver copper concentrations of growing Goslings from 28 to 70 d of age. Two hundred healthy male Yangzhou geese with similar body weight were randomized to four groups with five replicates per treatment and ten geese per replicate. Average daily feed intake, average daily gain and feed conversion ratio of geese for each pen were measured from 28 to 70 d of age. At 70 d of age, two geese were selected randomly from each pen and slaughtered to evaluate carcass quality. Metabolism experiment was conducted with five male geese from each group (one goose per pen) which body weight was close to the mean weight of the group from 64 to 70 d of age. Significant effects of dietary copper was found on body weight, feed conversion ratio, carcass yield, fecal copper concentrations and liver copper concentrations. Body weight, feed conversion ratio and carcass yield showed significant quadratic response to increase dietary copper concentration, while fecal copper concentration and liver copper concentration showed a significant linear response. The result showed that dietary Cu addition can improve growth by increasing the use of the feeding stuff and improving carcass yield in growing Goslings. Furthermore, taking into consideration, the optimal level of Gosling dietary copper was between 8.77 and 11.6 mg/kg from 28 to 70 days of age.
Assuntos
Animais , Recém-Nascido , Abate de Animais , Cobre/análise , Gansos/anormalidades , Gansos/fisiologia , Fezes/químicaRESUMO
Colocasia esculenta cv. Xinmaoyu is an eddoe-type taro cultivar local to Taicang, Jiangsu Province, China; it is characterized by its pure flavor, glutinous texture, and high nutritional value. Due to its excellent qualities, the Trademark Office of the State Administration for Industry and Commerce of the People's Republic of China awarded Xinmaoyu, a geographical indication certification in 2014. Therefore, there is an urgent need to develop an efficient molecular marker for the specific identification of this cultivar, which would greatly facilitate the conservation and utilization of this unique germplasm resource. In the present study, amplifying the psbE-petL fragment from two dasheen-type and seven eddoe-type taro cultivars revealed three conserved insertions/deletions among sequences from the two taro types. Based on these sequence differences, a pair of site-specific primers was designed targeting the psbE-petL sequence from the dasheen-type taro, which specifically amplified a DNA band in all individuals from cultivars of this type, but not in those from the seven eddoe-type cultivars. To discriminate Xinmaoyu from the other eddoe-type taro cultivars, a pair of simple sequence repeat-sequence characterized amplified region (SSR-SCAR) primers was further developed to specifically amplify a DNA band from all Xinmaoyu individuals, but not from individuals of other eddoe-type taro cultivars. In conclusion, through a two-step-screening procedure using psbE-petL and SSR-SCAR markers, we developed a pair of primers that could specifically discriminate Xinmaoyu from nine taro cultivars commonly cultivated in Jiangsu Province and Fujian Province.
Assuntos
Colocasia/genética , Marcadores Genéticos , Repetições de Microssatélites , Colocasia/classificação , HumanosRESUMO
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.
Assuntos
Síndrome de Klinefelter/genética , Proteínas de Plasma Seminal/genética , Azoospermia/genética , Deleção Cromossômica , Cromossomos Humanos Y/genética , Deleção de Genes , Loci Gênicos/genética , Humanos , Infertilidade Masculina/genética , Masculino , Fenótipo , Sitios de Sequências Rotuladas , Aberrações dos Cromossomos Sexuais , Transtornos do Cromossomo Sexual no Desenvolvimento Sexual/genéticaRESUMO
This study investigated the effects of pregnant mare se-rum gonadotropin (PMSG) and cloprostenol (CLO) on estrus induc-tion and synchronization, uterine development, and follicle-stimulating hormone receptor (FSHR) expression in mice. A total of 105 Kunming pre-puberty mice were divided into seven subgroups. Three PMSG sub-groups were injected intraperitoneally with 10, 20, and 40 IU PMSG twice (on days 0 and 4), and three CLO subgroups were injected intra-peritoneally with 10, 15, and 20 µg cloprostenol acetate twice (on days 0 and 4). The results showed that 93.33 and 66.67% of synchronized mice displayed estrus within 18.68-37.59 h following CLO and PMSG exposure, respectively. Estrus numbers, estrus onset time, and estrus rates in CLO and PMSG groups were greater than in control groups (CG) (P < 0.05). Uterine weights of the PMSG group were higher than that of CLO and CG groups, and the uterine horn longitudinal diameters in experimental mice were greater than CG. Expression levels of FSHR proteins in CLO and PMSG groups increased slightly when compared to CG. In conclusion, CLO and PMSG administration did not clearly af-fect the expression of uterine FSHR proteins in mice. Moreover, PMSG and CLO treatments synchronized estrus and enhanced the uterine de-velopment of mice. The efficacy of CLO on estrus synchronization was greater than PMSG, and the effects of PMSG on uterine development were stronger than CLO. These results have important significance re-garding the modulation of animal reproductive functions.
Assuntos
Cloprostenol/farmacologia , Estro/efeitos dos fármacos , Gonadotropinas Equinas/farmacologia , Luteolíticos/farmacologia , Receptores do FSH/genética , Útero/efeitos dos fármacos , Animais , Estro/fisiologia , Feminino , Expressão Gênica/efeitos dos fármacos , Cavalos , Injeções Intraperitoneais , Camundongos , Tamanho do Órgão/efeitos dos fármacos , Gravidez , Receptores do FSH/metabolismo , Maturidade Sexual/efeitos dos fármacos , Maturidade Sexual/fisiologia , Útero/crescimento & desenvolvimento , Útero/metabolismoRESUMO
This study investigated the in vitro and in vivo antiproliferative activity of esculetin against hepatocellular carcinoma, and clarified its potential molecular mechanisms. Cell viability was determined by the MTT (tetrazolium) colorimetric assay. In vivo antitumor activity of esculetin was evaluated in a hepatocellular carcinoma mouse model. Seventy-five C57BL/6J mice were implanted with Hepa1-6 cells and randomized into five groups (n=15 each) given daily intraperitoneal injections of vehicle (physiological saline), esculetin (200, 400, or 700 mg·kg-1·day-1), or 5-Fu (200 mg·kg-1·day-1) for 15 days. Esculetin significantly decreased tumor growth in mice bearing Hepa1-6 cells. Tumor weight was decreased by 20.33, 40.37, and 55.42% with increasing doses of esculetin. Esculetin significantly inhibited proliferation of HCC cells in a concentration- and time-dependent manner and with an IC50 value of 2.24 mM. It blocked the cell cycle at S phase and induced apoptosis in SMMC-7721 cells with significant elevation of caspase-3 and caspase-9 activity, but did not affect caspase-8 activity. Moreover, esculetin treatment resulted in the collapse of mitochondrial membrane potential in vitro and in vivo accompanied by increased Bax expression and decreased Bcl-2 expression at both transcriptional and translational levels. Thus, esculetin exerted in vitro and in vivo antiproliferative activity in hepatocellular carcinoma, and its mechanisms involved initiation of a mitochondrial-mediated, caspase-dependent apoptosis pathway.
Assuntos
Adulto , Feminino , Humanos , Masculino , Esgotamento Profissional/genética , Doenças em Gêmeos/genética , Local de Trabalho , Esgotamento Profissional/epidemiologia , Esgotamento Profissional/etiologia , Esgotamento Profissional/psicologia , Demografia , Doenças em Gêmeos/epidemiologia , Doenças em Gêmeos/etiologia , Doenças em Gêmeos/psicologia , Interação Gene-Ambiente , Sistema de Registros , Fatores de Risco , Inquéritos e Questionários , Suécia/epidemiologiaRESUMO
This study investigated the in vitro and in vivo antiproliferative activity of esculetin against hepatocellular carcinoma, and clarified its potential molecular mechanisms. Cell viability was determined by the MTT (tetrazolium) colorimetric assay. In vivo antitumor activity of esculetin was evaluated in a hepatocellular carcinoma mouse model. Seventy-five C57BL/6J mice were implanted with Hepa1-6 cells and randomized into five groups (n=15 each) given daily intraperitoneal injections of vehicle (physiological saline), esculetin (200, 400, or 700 mg·kg-1·day-1), or 5-Fu (200 mg·kg-1·day-1) for 15 days. Esculetin significantly decreased tumor growth in mice bearing Hepa1-6 cells. Tumor weight was decreased by 20.33, 40.37, and 55.42% with increasing doses of esculetin. Esculetin significantly inhibited proliferation of HCC cells in a concentration- and time-dependent manner and with an IC50 value of 2.24 mM. It blocked the cell cycle at S phase and induced apoptosis in SMMC-7721 cells with significant elevation of caspase-3 and caspase-9 activity, but did not affect caspase-8 activity. Moreover, esculetin treatment resulted in the collapse of mitochondrial membrane potential in vitro and in vivo accompanied by increased Bax expression and decreased Bcl-2 expression at both transcriptional and translational levels. Thus, esculetin exerted in vitro and in vivo antiproliferative activity in hepatocellular carcinoma, and its mechanisms involved initiation of a mitochondrial-mediated, caspase-dependent apoptosis pathway.
Assuntos
Antioxidantes/uso terapêutico , Apoptose/efeitos dos fármacos , Carcinoma Hepatocelular/tratamento farmacológico , Neoplasias Hepáticas/tratamento farmacológico , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Umbeliferonas/uso terapêutico , Animais , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Neoplasias Experimentais , Distribuição AleatóriaRESUMO
The specific role of Toll-like receptor 4 (TLR4) in bleomycin-induced lung fibrosis of mice, a model of human idiopathic pulmonary fibrosis, has not been characterized. We injected bleomycin intratracheally into TLR4 knockout (TLR4(-/-)) and wild-type (WT) mice. Twenty-one days after injection, mice were sacrificed and their lungs were harvested for pathological, hydroxyproline, mRNA expression, and collagen I analyses. Body weight changes and mortality were observed. Light microscopy showed that lung fibrosis was minimal in TLR4(-/-) compared to that in WT mice on day 21 after bleomycin instillation. The Ashcroft score was significantly lower in TLR4(-/-) than in WT mice (3.667 ± 0.730 vs 4.945 ± 0.880, P < 0.05). Hydroxyproline content was significantly lower in TLR4(-/-) than in WT mice on day 21 after bleomycin injection (0.281 ± 0.022 vs 0.371 ± 0.047, P < 0.05). Compared to WT mice, bleomycin-treated TLR4(-/-) mice expressed significantly lower type I collagen mRNA levels (mesenchymal marker; 11.069 ± 2.627 vs 4.589 ± 1.440, P < 0.05). Collagen I was significantly lower in TLR4(-/-) than in WT mice (0.838 ± 0.352 vs 2.427 ± 0.551, P < 0.05). Bleomycin-treated TLR4(-/-) mice had a significantly lower mortality rate on day 21 than WT mice (33 vs 75%, P < 0.05). Body weight reduction was lower in TLR4(-/-) mice than in WT mice; this difference was not statistically significant (-3.735 ± 5.276 vs -6.698 ± 3.218, P > 0.05). Thus, bleomycin-induced pulmonary fibrosis is TLR4-dependent and TLR4 promoted fibrosis in bleomycin-challenged mice.
Assuntos
Fibrose/genética , Lesão Pulmonar/genética , Receptor 4 Toll-Like/genética , Animais , Bleomicina/toxicidade , Colágeno Tipo I/biossíntese , Fibrose/induzido quimicamente , Fibrose/patologia , Humanos , Lesão Pulmonar/patologia , Camundongos , Camundongos Knockout , RNA Mensageiro/biossínteseRESUMO
DNA markers are useful tools that play an important role in plant cultivar identification. They are usually based on polymerase chain reaction (PCR) and include simple sequence repeats (SSRs), inter-simple sequence repeats, and random amplified polymorphic DNA. However, DNA markers were not used effectively in the complete identification of plant cultivars because of the lack of known DNA fingerprints. Recently, a novel approach called the cultivar identification diagram (CID) strategy was developed to facilitate the use of DNA markers for separate plant individuals. The CID was designed whereby a polymorphic maker was generated from each PCR that directly allowed for cultivar sample separation at each step. Therefore, it could be used to identify cultivars and varieties easily with fewer primers. In this study, 60 apple cultivars, including a few main cultivars in fields and varieties from descendants (Fuji x Telamon) were examined. Of the 20 pairs of SSR primers screened, 8 pairs gave reproducible, polymorphic DNA amplification patterns. The banding patterns obtained from these 8 primers were used to construct a CID map. Each cultivar or variety in this study was distinguished from the others completely, indicating that this method can be used for efficient cultivar identification. The result contributed to studies on germplasm resources and the seedling industry in fruit trees.
Assuntos
Malus/classificação , Malus/genética , Repetições de Microssatélites , Marcadores Genéticos , Polimorfismo GenéticoRESUMO
Rats with unilateral 6-hydroxydopamine (6-OHDA) lesions were exposed to L-DOPA treatments, 25 mg kg-1 L-DOPA methyl ester/2 mg kg-1 carbidopa, or to saline. Fourteen days later, both sets of animals were tested with the L-DOPA/carbidopa treatment. The L-DOPA pre-exposed animals exhibited behavioral sensitization as indexed by a higher frequency of contralateral rotations. Although striatal L-DOPA and HVA concentrations were equivalent in the two groups, the L-DOPA treatment induced a 10 fold variation in the range of L-DOPA concentrations in the 6-OHDA striatum in both sets of animals. Importantly, significant correlations between striatal L-DOPA concentration and behavior were obtained only for the L-DOPA pre-exposed animals.
Assuntos
Comportamento Animal/efeitos dos fármacos , Levodopa/farmacologia , Neostriado/metabolismo , Animais , Química Encefálica/efeitos dos fármacos , Dopamina/metabolismo , Ácido Homovanílico/metabolismo , Levodopa/farmacocinética , Masculino , Neostriado/efeitos dos fármacos , Oxidopamina/farmacologia , Ratos , Ratos Sprague-Dawley , Técnicas EstereotáxicasRESUMO
Rats with severe unilateral dopamine denervation (> or = 95% dopamine deficit), produced by intracerebral injection of 6-hydroxydopamine (6-OHDA) into the ventral tegmentum nigrostriatal dopamine neurons, were administered 25 mg/kg L-dopa (3,4-dihydroxyphenylalanine) methyl ester/2 mg/kg carbidopa. The neurochemical effects of the L-dopa treatment on uric acid in the cortex and striatum of the intact and 6-OHDA hemisphere were measured. In comparison to saline animals, uric acid concentrations in brain were increased in the L-dopa treated animals. There were no interhemispheric differences in the uric acid concentrations either in the L-dopa or in the saline treated animals. Interhemispheric differences were, however, observed in terms of the correlations obtained between L-dopa and uric acid concentrations in the intact vs. the 6-OHDA hemisphere. Statistically significant correlation coefficients were found in the striatal and cortex samples obtained from the 6-OHDA hemisphere. Furthermore, high correlation coefficients were observed between contralateral rotation frequencies and uric acid concentrations in the cortex and striatum of the 6-OHDA hemisphere. In contrast, only low and statistically non significant correlations were observed in the tissue samples obtained from the intact hemisphere. These observations suggest that L-dopa activation of the dopamine supersensitive receptors of the DA denervated hemisphere and the associated metabolism of purines with high energy phosphate bonds (e.g. ATP and GTP) increases uric acid as an end-product of purine metabolism. These findings are consistent with other findings indicating that uric acid in the brain can provide an index of metabolic activation in brain tissue.