RESUMEN
OBJECTIVE: To explore the specific mechanism of sevoflurane in alleviating cerebral ischemia-reperfusion injury (CIRI) in rats through the c-Jun N-terminal kinase (JNK) signaling pathway. MATERIALS AND METHODS: A total of 60 male specific pathogen-free Sprague-Dawley rats were randomly divided into sham group (n=20), model group (n=20), and sevoflurane group (n=20). In the sevoflurane group, sevoflurane (2.5%) was inhaled for 60 min at 24 h before the blockage of cerebral blood supply. The CIRI model was established using the suture method in the model group and sevoflurane group, while the right common carotid artery and external carotid artery were separated and ligated only, without suture placement, in the sham group. At 24 h after reperfusion, the neurological deficit score in each group was calculated, the water content in brain tissues in each group was detected based on dry-wet weight ratio, the infarction volume of brain tissues in each group was detected via 2,3,5-triphenyltetrazolium chloride (TTC) staining, and the apoptosis rate of brain cells in each group was detected using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. Moreover, the protein levels of JNK, p-JNK, B-cell lymphoma-2 (Bcl-2), and the Bcl-2 associated X protein (Bax) in brain tissues were determined using Western blotting, and the gene expressions of Bax and Bcl-2 in brain tissues were determined through fluorescence quantitative Polymerase Chain Reaction (qPCR). RESULTS: It was found that the water content in brain tissues and the cerebral infarction volume were significantly increased in the model group compared with those in the sham group (p<0.01, p<0.01), while they were notably decreased in the sevoflurane group compared with those in the model group (p<0.05, p<0.01). The neurological deficit score was significantly higher in the model group than that in the sham group (p<0.01), while it was remarkably lower in the sevoflurane group than that in the model group (p<0.01). According to the results of the TUNEL assay, the model group had an evidently higher apoptosis rate of brain cells than the sham group (p<0.01), while the sevoflurane group had a lower apoptosis rate of brain cells than the model group (p<0.05). Besides, the results of Western blotting revealed that the model group exhibited remarkably increased protein levels of JNK, p-JNK, and Bax (p<0.05, p<0.01, p<0.01) and a remarkably decreased protein level of Bcl-2 (p<0.01) compared with the sham group. Sevoflurane group had decreased protein levels of JNK, p-JNK, and Bax (p<0.05, p<0.01, p<0.01) and an increased protein level of Bcl-2 (p<0.05) in comparison with the model group. In addition, the gene expression of Bcl-2 significantly declined (p<0.01), and that of Bax remarkably rose (p<0.01) in the model group compared with those in the sham group, while the contrary is the case in the sevoflurane group compared with those in the model group (p<0.05, p<0.01). CONCLUSIONS: Sevoflurane can regulate the protein and gene expressions of Bax and Bcl-2 and reduce apoptosis in CIRI by regulating the JNK signaling pathway, thereby exerting a protective effect on brain tissues and improving the symptoms of neurological deficit.
Asunto(s)
Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Daño por Reperfusión/tratamiento farmacológico , Sevoflurano/farmacología , Transducción de Señal/efectos de los fármacos , Animales , Apoptosis/efectos de los fármacos , Masculino , Ratas , Ratas Sprague-Dawley , Daño por Reperfusión/metabolismo , Daño por Reperfusión/patologíaRESUMEN
OBJECTIVE: To observe the expressions of Linc-ROR and proteins in the PI3K-Akt pathway in an ectopic lesion of adenomyosis. PATIENTS AND METHODS: The expression of Linc-ROR in the ectopic endometrium, eutopic endometrium, and normal endometrium of adenomyosis was detected by qRT-PCR. Western blot was used to detect the protein expressions of PI3K-Akt in endometriosis and lesion endometriosis. Cell counting kit-8 (CCK-8) assay was utilized to detect cell proliferative activity. After interfering or overexpressing Linc-ROR, protein expressions of the PI3K-Akt pathway were detected by Western blot. RESULTS: Linc-ROR expression in the ectopic endometrium of adenomyosis was higher than that in the eutopic endometrium and normal endometrium, and the expression level of PTEN in adenomyosis tissues was decreased, whilst expression levels of Akt, p-Akt, p-PTEN were increased. Clinical data of enrolled patients indicated that there was a relationship between Linc-ROR expression and the type and severity of dysmenorrhea of adenomyosis. However, no relationship was observed between Linc-ROR expression and age, cesarean section, uterine surgery, and menstrual cycle. Cell counting kit-8 (CCK-8) assay showed that the proliferative activity of cells was significantly decreased after knockdown of Linc-ROR in the adenomyosis cells. Western blot revealed that the expression level of PTEN increased but the expression levels of p-Akt, p-PTEN and p-PDK1 decreased. Overexpression of Linc-ROR obtained the opposite results. CONCLUSIONS: Linc-ROR is highly expressed in the ectopic endometrium of adenomyosis, and it can promote the proliferative activity of endometrial cells by activating the PI3K-Akt pathway.
Asunto(s)
Endometrio/metabolismo , Fosfatidilinositol 3-Quinasas/fisiología , Proteínas Proto-Oncogénicas c-akt/fisiología , ARN Largo no Codificante/fisiología , Transducción de Señal/fisiología , Adenomiosis/metabolismo , Adulto , Proliferación Celular , Endometriosis/metabolismo , Endometrio/citología , Femenino , Humanos , Fosfohidrolasa PTEN/fisiología , EmbarazoRESUMEN
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.
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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 , EmbarazoRESUMEN
We investigated the expression of salivary α2-macroglobulin (α2-MG) in patients with type 2 diabetes mellitus (T2DM) to investigate its value for predicting damage to the salivary glands. A total of 116 patients with T2DM and 60 patients with impaired fasting glucose (IFG) were included in this study. Sixty health volunteers were enrolled as a control group. Unstimulated saliva was collected at 8 a.m. prior to breakfast. Expression of α2-MG was determined using an enzyme-linked immunosorbent assay. The correlation between salivary α2-MG, serum α2-MG, and concentration of fasting glucose was analyzed using Pearson correlation analysis. No significant difference was observed in the expression of serum α2-MG in the T2DM group, IFG group, and control group (P > 0.05). Compared with the control group and IFG group, a statistical difference was observed in the salivary α2-MG in the T2DM group (P < 0.01). No statistical difference was observed in the salivary α2-MG in the IFG group compared with the control group (P > 0.05). In the patients with T2DM, a close correlation was identified in the expression of serum α2-MG and salivary α2-MG (r = 0.52, P < 0.01). A poor correlation was identified between salivary α2-MG and blood sugar level (r = -0.12, P = 0.199). The expression of salivary α2-MG showed a remarkable increase in T2DM patients, which may be associated with functional disorders of the salivary gland.
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Diabetes Mellitus Tipo 2/metabolismo , Saliva/metabolismo , Regulación hacia Arriba , alfa-Macroglobulinas/metabolismo , Anciano , Análisis de Varianza , Glucemia/metabolismo , Diabetes Mellitus Tipo 2/sangre , Ensayo de Inmunoadsorción Enzimática , Ayuno/sangre , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
This study investigated cadherin-1 (Cdh1) expression in the sensorimotor cortex of rats after spinal cord injury (SCI). The repairing effect of Cdh1 was evaluated by silencing its expression with lentivirus-mediated RNAi. Twenty male Sprague-Dawley (SD) rats were randomly divided into a normal group and an operation group. Rats of the operation group were given SCI by the Allen method (T10-T11). Cdh1 expression in the sensorimotor cortex was examined by quantitative real-time polymerase chain reaction (PCR) and Western blot analysis. Thirty male SD rats were divided into a sham-operation (SO) group, a lentivirus vector (LV) group, and a recombinant lentivirus (RL) group. Rat behavior was evaluated using the Basso-Beattie-Bresnahan (BBB) test every week. Ten days after injection, Cdh1 expression was examined by quantitative real-time PCR and Western blot. Six weeks after injury, animals were injected with biotinylated dextran amine-Texas Red (BDA-TR), and then at 8 weeks, spinal cords were removed and sectioned in serial order. The expression of Cdh1 mRNA was significantly higher in the operation than in the normal group (P < 0.05). The expression of Cdh1 mRNA was lower in the RL than in the SO or LV groups at 10 days after injection (P < 0.05). In addition, the BBB score was higher for the RL than for the SO or LV groups at 6 weeks after injury (P < 0.05). A novel population of BDA-labeled axons was observed extending past the lesion in the RL group, which was rarely observed in the SO and LV groups. These results suggest that the anaphase-promoting complex-Cdh1 may play an important role in inhibiting axonal growth.
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Proteínas Cdh1/genética , Regulación de la Expresión Génica , Interferencia de ARN , Traumatismos de la Médula Espinal/genética , Animales , Axones/metabolismo , Conducta Animal , Proteínas Cdh1/metabolismo , Modelos Animales de Enfermedad , Técnica del Anticuerpo Fluorescente , Expresión Génica , Vectores Genéticos/genética , Lentivirus/genética , Masculino , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas , Traumatismos de la Médula Espinal/metabolismoRESUMEN
We investigated the effects of cadmium on lung cell DNA in immature mice. The mice were randomly divided into four groups: control group, low-dose group (1/100 LD(50)), middle-dose group (1/50 LD(50)), and high-dose group (1/25 LD(50)); they were supplied with cadmium chloride or control water for 40 days. Lung cells collected from sacrificed mice were used to evaluate the extent of DNA damage by comet assay. The ratio of tailing cells, DNA tail length, DNA comet length, DNA tail moment, DNA olive tail moment, and percentage of DNA in the comet tail were measured. The rate of tailing lung cells exposed to cadmium increased significantly; the low-concentration group had significantly (P < 0.05) higher rates, and the middle- and high-concentration groups had higher (P < 0.01) rates compared to the control. DNA tail length, DNA comet length, DNA tail moment, and DNA olive tail moment all increased with the increase in cadmium doses, but compared with those of the control group, no significant differences in low-dose group were found (P > 0.05), and the differences in middle- and high-dose groups were all highly significant (P < 0.01). The degree of DNA damage also increased with the increase of the cadmium concentrations. We conclude that cadmium significantly increases DNA damage in lung cells of immature mice in a dose-dependent manner.
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Cloruro de Cadmio/toxicidad , Daño del ADN , Agua Potable/efectos adversos , Pulmón/patología , Contaminantes Químicos del Agua/toxicidad , Animales , Ensayo Cometa , Femenino , Dosificación Letal Mediana , Pulmón/efectos de los fármacos , Masculino , RatonesRESUMEN
Vero cell was selected as productive carrier in production of Attenuated IBDV. The optimal conditions in culturing IBDV in agitating bottle were figured out. After proliferation, supernatant was harvested in batch process under the condition of five-liter-agitated fermenter. The result shows it was successful to culture IBDV by this methodology.
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Virus de la Enfermedad Infecciosa de la Bolsa/crecimiento & desarrollo , Cultivo de Virus , Animales , División Celular , Chlorocebus aethiops , Células VeroRESUMEN
282 cervical swab specimens and serum samples collected from pregnant women were used for virological studies in order to investigate the carrier status in herpes viridae and antibody. Seven isolates were obtained with a positive rate of 2.48%. According to the characteristics of cytopathology effects on the diploid cells (2BS), pathogenicity in the hosts and electron micrographic findings, these 7 isolates belonged to herpes viridae. The results of neutralization test, enzyme linked immunosorbent assay blocking test and indirect immunofluorescence tests against HCMV-specific serum and standard HCMV AD-169, HSV-ISm44, HSV-IIG indicated that all these 7 viruses were HCMV. No other viral members of herpes viridae were detected in this study.
Asunto(s)
Portador Sano/microbiología , Cuello del Útero/microbiología , Infecciones por Citomegalovirus/microbiología , Herpes Simple/microbiología , Complicaciones Infecciosas del Embarazo/microbiología , Citomegalovirus/aislamiento & purificación , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Embarazo , Simplexvirus/aislamiento & purificaciónRESUMEN
Seven continuous primate cell lines were tested in three systems (nude mice, muscle organ culture, and soft agarose) for their ability to express characteristics usually associated with malignant cell lines. Five of the seven cell lines failed to produce tumors in nude mice, failed to show a tumor-like pattern of growth in muscle organ culture, and failed to produce colonies in soft agarose. The remaining two cell lines showed different degrees of tumorigenicity in nude mice, and gave frankly positive results in the two in vitro assays. In addition, one of these lines appeared to progress from potential to overt tumorigenicity. We conclude that acquisition of infinite life in primate cell lines is not invariably equivalent to the ability to form tumors.
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Línea Celular , Transformación Celular Neoplásica , Animales , Chlorocebus aethiops , Femenino , Humanos , Macaca mulatta , Ratones , Ratones Desnudos , Técnicas de Cultivo de ÓrganosRESUMEN
36 young adult volunteers were given either virus, or inactivated surface-antigen influenza vaccine, containing influenza A (H1N1) antigen. Their peripheral-blood lymphocytes were analysed to determine whether cytotoxic T lymphocytes (CTL) were induced by these vaccines. All three vaccines induced HLA-restricted T-lymphocyte responses specific for influenza A virus. This CTL response was found in 28 of 30 volunteers who showed antibody responses and in 3 of 6 who did not develop antibodies. Virus infected target cells showing certain HLA antigens (A1, A9, and B8) were less susceptible to lysis by HLA-matched cytotoxic T effector cells than target cells which matched effectors at other HLA loci.