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1.
J Biol Regul Homeost Agents ; 34(2): 525-533, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32425017

RESUMO

To explore effects of the sDR5-Fc fusion protein on ulcerative colitis of infant mice via the TRAIL-DR5 pathway, 50 female mice were randomly divided into 5 groups, i.e., control group (group A), dextran sulfate sodium group (group B), hIgG group (group C), 10 mg/kg sDR5-Fc group (group D), and 20 mg/ kg sDR5-Fc group (group E). The acute ulcerative colitis models were established. The weights and disease activity index (DAI) of each group were monitored daily. In addition, the pathological changes of colon tissues were observed by Hematoxylin-Eosin staining. The number of macrophages in colon tissues was detected by immunohistochemistry assay. Changes in the expression of inflammatory factors in colon tissues were detected by quantitative real-time polymerase chain reaction (PCR). Lipopolysaccharide (LPS) of different concentrations was utilized alone or in combination with TRAIL to stimulate the NCM460 cells. The activation of NLRP3 inflammasomes was detected by Western blot. The apoptosis of NCM460 cells was detected by flow cytometry. The results showed that in groups B and C, the body weights decreased, the DAI increased, the colon epithelial cells were injured, the inflammatory cells were infiltrated, and the macrophages in colon tissues increased significantly. In groups D and E, the body weights increased, the DAI decreased, the inflammation was significantly improved, the macrophages decreased significantly, and the gene expression levels of NLRP3, Caspase-1, and IL-1ß decreased significantly. Thus, sDR5-Fc could inhibit the activation of NLRP3 inflammasomes induced by TRAIL, thereby decreasing the apoptosis of NCM460 cells. In conclusion, the sDR5-Fc fusion protein could block the TRAIL-DR5 pathway to reduce the expression of NLRP3 inflammasomes, thereby improving ulcerative colitis.

2.
Sci Rep ; 9(1): 19173, 2019 12 16.
Artigo em Inglês | MEDLINE | ID: mdl-31844123

RESUMO

Extensive research has revealed the association of continued oxidative stress with chronic inflammation, which could subsequently affect many different chronic diseases. The mycotoxin deoxynivalenol (DON) frequently contaminates cereals crops worldwide, and are a public health concern since DON ingestion may result in persistent intestinal inflammation. There has also been considerable attention over the potential of DON to provoke oxidative stress. In this study, the cytoprotective effect of Schisandrin A (Sch A), one of the most abundant active dibenzocyclooctadiene lignans in the fruit of Schisandra chinensis (Turcz.) Baill (also known as Chinese magnolia-vine), was investigated in HT-29 cells against DON-induced cytotoxicity, oxidative stress and inflammation. Sch A appeared to protect against DON-induced cytotoxicity in HT-29 cells, and significantly lessened the DON-stimulated intracellular reactive oxygen species and nitrogen oxidative species production. Furthermore, Sch A lowered DON-induced catalase, superoxide dismutase and glutathione peroxidase antioxidant enzyme activities but maintains glutathione S transferase activity and glutathione levels. Mechanistic studies suggest that Sch A reduced DON-induced oxidative stress by down-regulating heme oxygenase-1 expression via nuclear factor (erythroid-derived 2)-like 2 signalling pathway. In addition, Sch A decreased the DON-induced cyclooxygenase-2 expression and prostaglandin E2 production and pro-inflammatory cytokine interleukin 8 expression and secretion. This may be mediated by preventing DON-induced translocation of nuclear factor-κB, as well as activation of mitogen-activated protein kinases pathways. In the light of these findings, we concluded that Sch A exerted a cytoprotective role in DON-induced toxicity in vitro, and it would be valuable to examine in vivo effects.

3.
Eur Rev Med Pharmacol Sci ; 23(24): 10761-10768, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31858543

RESUMO

OBJECTIVE: To uncover the biological effect of long non-coding RNA (lncRNA) PAPAS on the progression of gastric cancer (GC) by mediating microRNA-188-5p (miRNA-188-5p) level. PATIENTS AND METHODS: The relative level of PAPAS was determined in GC tissues and cell lines by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The Kaplan-Meier method was introduced to assess the prognostic potential of PAPAS in the overall survival of GC patients. Regulatory effects of PAPAS on proliferative, migratory, and invasive abilities of HGC-27 and AGS cells were detected by cell counting kit-8 (CCK-8), transwell, and wound closure assay, respectively. Subsequently, the binding relation between PAPAS and miRNA-188-5p was verified by the Dual-luciferase reporter gene assay. Correlation between expression levels of PAPAS and miRNA-188-5p in GC tissues was explored. Finally, rescue experiments were conducted to uncover the role of PAPAS/miRNA-188-5p axis in the progression of GC. RESULTS: PAPAS was upregulated in GC tissues and cell lines compared to controls. GC patients expressing a high level of PAPAS suffered worse prognosis relative to those with low level. The silence of PAPAS remarkably attenuated proliferative, migratory, and invasive abilities of HGC-27 cells. Overexpression of PAPAS in AGS cells obtained the opposite trends. MiRNA-188-5p was the direct target of PAPAS, which was negatively regulated by PAPAS. MiRNA-188-5p was able to reverse the regulatory effects of PA-PAS on proliferative, migratory, and invasive abilities of GC cells. CONCLUSIONS: LncRNA PAPAS is upregulated in GC and closely related to lymphatic metastasis, distant metastasis, and poor prognosis of GC patients. PAPAS aggravate the malignant progression of GC by negatively regulating the miRNA-188-5p level.

4.
Eur Rev Med Pharmacol Sci ; 23(24): 10769-10775, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31858544

RESUMO

OBJECTIVE: To uncover the function of LINC00461 in regulating cellular behaviors of gastric cancer (GC) via targeting LSD1. PATIENTS AND METHODS: LINC00461 level in GC tissues with different tumor node metastasis (TNM) staging and lymphatic metastasis statues was determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). In vitro influences of LINC00461 on proliferative and apoptotic rates were evaluated in AGS and SGC-7901 cells. The interaction between LINC00461 and LSD1 was explored by RNA immunoprecipitation (RIP) assay and qRT-PCR. Finally, the potential role of LSD1 in the proliferative ability of GC cells mediated by LINC00461 was assessed. RESULTS: LINC00461 level was higher in GC tissues relative to matched control ones. It was positively correlated to TNM staging and lymphatic metastasis of GC. Knockdown of LINC00461 markedly attenuated viability and the proliferative ability of AGS and SGC-7901 cells, but induced apoptosis. RIP assay demonstrated the interaction between LINC00461 and LSD1. Moreover, LSD1 could reverse the regulatory effect of LINC00461 on the proliferative ability of GC cells. CONCLUSIONS: LINC00461 is upregulated in GC, which is positively related to TNM staging and lymphatic metastasis. LINC00461 mediates proliferation and apoptosis of GC cells, thereafter aggravating the progression of GC.

5.
J Food Sci ; 84(7): 1854-1863, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31206699

RESUMO

Lactobacillus rhamnosus strain ASCC 1520 with high soy isoflavone transformation ability was used to ferment soymilk and added to the diet of mice. The impact of L. rhamnosus fermentation on soy isoflavone metabolites and intestinal bacterial community, in conjunction with fecal enzyme activity and short-chain fatty acids (SCFA) excretion was evaluated. Antibiotics intervention resulted in a decrease in fecal enzyme activities and SCFA. Although long-term intake of soymilk or L. rhamnosus-fermented soymilk did not affect the fecal ß-glucuronidase and ß-galactosidase activities, it improved the ß-glucosidase activity when antibiotics were concomitantly administered. Soymilk or fermented soymilk administration increased the isoflavone metabolites (O-DMA and equol) excreted in urine. Antibiotics decreased the daidzein excretion and its metabolites but showed little effect on glycitein and genistein excretion. Principal coordinates analysis (PCoA) of the 16s rRNA gene sequencing data found a remarkable shift in gut microbiota after soymilk administration and antibiotics treatment. Matastats test of the relative abundance of bacterial taxa revealed Odoribacter (Bacteroidales family), Lactobacillus (Lactobacillales order), and Alistipes (Rikenellaceae family) were enriched in soymilk while bacterial taxa from Bacteroides and Lactobacillus were enriched in L. rhamnosus-fermented soymilk. Furthermore, there was less decrease in bacterial taxa with fermented soymilk group even when antibiotics were concomitantly administered. Overall, this study revealed that the gut microbiota of a healthy host is enough for the whole isoflavone metabolism under normal conditions. Feeding mice with L. rhamnosus-fermented soymilk improved fecal enzyme activity and kept the balance of the gut mirobiota when antibiotics were used. PRACTICAL APPLICATION: Feeding mice with L. rhamnosus-fermented soymilk improved fecal enzyme activity and kept the balance of the gut mirobiota when antibiotics were used.


Assuntos
Ácidos Graxos Voláteis/metabolismo , Fezes/microbiologia , Microbioma Gastrointestinal , Isoflavonas/metabolismo , Lactobacillales/metabolismo , Leite de Soja/metabolismo , Animais , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Bactérias/metabolismo , Fermentação , Microbiologia de Alimentos , Genisteína/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , beta-Glucosidase/metabolismo
6.
Zhonghua Gan Zang Bing Za Zhi ; 27(4): 250-255, 2019 Apr 20.
Artigo em Chinês | MEDLINE | ID: mdl-31082334

RESUMO

Objective: To investigate the correlation between interleukin-6 (IL-6) single nucleotide polymorphism (SNP) and the occurrence and prognosis of hepatitis B virus-associated acute-on-chronic liver failure (HBV-ACLF). Methods: Patients with chronic hepatic diseases diagnosed as HBV infection in the Hepatology Center of the First Affiliated Hospital of Fujian Medical University from July 2012 to March 2018 were divided into HBV-ACLF and non-ACLF group. SNP genotyping of eight loci in IL-6 gene (rs1524107, rs1800795, rs1800797, rs2069827, rs2069830, rs2069837, rs2069840 and rs2069845) was determined by the improved multi-temperature ligase detection reaction (imLDRTM) technique. Simultaneously, case data were reviewed with the 3-months followed up survival condition of the ACLF group. Normally distributed data were expressed as arithmetic means and SDs, and t-test was adopted. Data with skewed distribution were expressed as medians with interquartile range, and were measured by non-parametric test. Multivariate logistic regression analysis was used to analyze the relative risk of genetic polymorphism and HBV-ACLF as well as the relationship between IL-6 SNPs with the occurrence and prognosis of HBV-ACLF. Results: Four hundred patients were included in the study, with 122 (30.5%) in the HBV-ACLF and 278 (69.5%) in the non-ACLF group. There were significant differences in total bilirubin, albumin, and white blood cell count, percentage of neutrophils, platelet count, alanine aminotransferase, aspartate aminotransferase, prothrombin time and international standardized ratio, creatinine and the model for end-stage liver disease score between the two groups (P < 0.001). The genotype of IL-6 genes (rs1800795, rs1800797, rs2069827, and rs2069830) of all subjects showed no mutation or the mutation rate under 1%. There was no significant difference in the genotype of IL-6 (rs1524107, rs2069837, rs2069840 and rs2069845) between the two groups (P > 0.05). Multivariate logistic regression analysis showed that the SNPs in the above four loci of IL-6 gene was not associated with HBV-ACLF risk, nor had significant correlation with the 3-months prognosis. Conclusion: The SNP genotyping of eight loci in IL-6 gene (rs1524107, rs1800795, rs1800797, rs2069827, rs2069830, rs2069837, rs2069840 and rs2069845) is unrelated to the occurrence and short-term prognosis of HBV-ACLF.


Assuntos
Insuficiência Hepática Crônica Agudizada/virologia , Vírus da Hepatite B/genética , Hepatite B Crônica/virologia , Hepatite B/virologia , Interleucina-6/genética , Insuficiência Hepática Crônica Agudizada/genética , DNA Viral/genética , Hepatite B Crônica/diagnóstico , Humanos , Polimorfismo de Nucleotídeo Único , Prognóstico
7.
Zhonghua Xue Ye Xue Za Zhi ; 40(2): 111-116, 2019 Feb 14.
Artigo em Chinês | MEDLINE | ID: mdl-30831625

RESUMO

Objective: To explore the effect of combination regimen of interferon alpha-1b, interleukin-2 and thalidomide (ITI regimen) on minimal residual disease (MRD) in patients with acute myeloid leukemia (AML) who were in hematologic remission but MRD-positive. Methods: Eighteen patients (17 from Tumor Hospital of Zhengzhou University and 1 from the First People's Hospital of Pingdingshan City) with AML admitted from July 2016 to June 2018, who were in hematologic remission but MRD-positive were treated with different doses of ITI regimen, and the MRD levels were monitored. Results: Among 18 patients who received a conventional dose of ITI regimen for 1 to 2 months, 7 patients had undetectable MRD, 3 had significant decrease in MRD levels, 3 had elevated MRD level and had hematologic recurrence. Three patients with elevated MRD level received a higher dose of ITI regimen, 2 of them turned to MRD negative and the other 1 patient had decreased MRD level. The total response rate was 72.2%, and the response rate in patients with MRD > 1.0% was 57.1% (4/7) , and that of patients with MRD < 1.0% was 81.8% (9/11) , respectively. Conclusion: The ITI regimen can reduce the MRD level of patient with AML who are in hematologic remission but MRD-positive. The therapeutic effect could be improved by a higher dose administration of ITI regimen, and therapeutic effect may be negatively correlated with MRD level before treatment.


Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Leucemia Mieloide Aguda , Citometria de Fluxo , Humanos , Interferon-alfa , Interleucina-2 , Leucemia Mieloide Aguda/tratamento farmacológico , Neoplasia Residual , Prognóstico , Indução de Remissão , Talidomida
8.
Crit Rev Food Sci Nutr ; 59(12): 1927-1936, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-29381385

RESUMO

Intestinal epithelial cells (IECs) lining the gastrointestinal tract establish a barrier between external environments and the internal milieu. An intact intestinal barrier maintains gut health and overall good health of the body by preventing from tissue injury, pathogen infection and disease development. When the intestinal barrier function is compromised, bacterial translocation can occur. Our gut microbiota also plays a fundamentally important role in health, for example, by maintaining intestinal barrier integrity, metabolism and modulating the immune system, etc. Any disruption of gut microbiota composition (also termed dysbiosis) can lead to various pathological conditions. In short, intestinal barrier and gut microbiota are two crucial factors affecting gut health. The gastrointestinal tract is a complex environment exposed to many dietary components and commensal bacteria. Dietary components are increasingly recognized to play various beneficial roles beyond basic nutrition, resulting in the development of the functional food concepts. Various dietary modifiers, including the consumption of live bacteria (probiotics) and ingestible food constituents such as prebiotics, as well as polyphenols or synbiotics (combinations of probiotics and prebiotics) are the most well characterized dietary bioactive compounds and have been demonstrated to beneficially impact the gut health and the overall well-being of the host. In this review we depict the roles of intestinal epithelium and gut microbiota in mucosal defence responses and the influence of certain functional food components on the modulation of gut health, with a particular focus on probiotics, prebiotics and polyphenols.


Assuntos
Alimento Funcional , Trato Gastrointestinal , Nível de Saúde , Dieta , Disbiose , Microbioma Gastrointestinal/imunologia , Trato Gastrointestinal/efeitos dos fármacos , Trato Gastrointestinal/imunologia , Trato Gastrointestinal/microbiologia , Humanos , Sistema Imunitário , Mucosa Intestinal/imunologia , Mucosa Intestinal/microbiologia , Intestinos/imunologia , Intestinos/microbiologia , Polifenóis/farmacologia , Prebióticos , Probióticos , Simbiose/imunologia
9.
Zhonghua Xue Ye Xue Za Zhi ; 39(5): 404-407, 2018 May 14.
Artigo em Chinês | MEDLINE | ID: mdl-29779350

RESUMO

Objective: To evaluate the efficacy and safety of a domestic human plasma derived coagulation Factor Ⅸ concentrate (pd-FⅨ) in patients with hemophilia B. Methods: The study was a multicenter, open-label and single-arm study. The efficacy of pd-F Ⅸ was evaluated by objective performance criteria. The doses of pd-FⅨ were calculated according to the bleeding symptom and disease severity. The infusion efficiency of pd-FⅨ and improvement of bleeding symptoms were measured at 30 minutes and (24±4) h after the first infusion, respectively. Adverse events were recorded. Viral infection and FⅨ inhibitor were detected 90 d after the first infusion. Results: All 36 subjects with hemophilia B were enrolled in the study. The median age of these patients was 31 years old and the median injection doses were 4 (1-17) times. The hemostatic effect of 27/36 (75.00%) and 9/36 (25.00%) acute bleeding events were rated as "excellent" and "better" , respectively. The recovery rate was 111.92% (65.55%-194.28%) at 30 minutes after infusion of FⅨ. There was no adverse event related to FⅨ. No reactivation of HBV, HCV or HIV and FⅨ inhibitor was detected at 90-104 d after the first FⅨ infusion. Conclusion: This domestically made human plasma derived FⅨ concentrate is safe and effective in the treatment of acute bleeding in patients with hemophilia B. Clinical trial registration: China food and Durg Administration, 2016L08027.


Assuntos
Hemofilia B , Adulto , China , Fator IX , Hemofilia A , Hemofilia B/terapia , Hemorragia , Humanos , Plasma
11.
Phys Chem Chem Phys ; 19(28): 18505-18513, 2017 Jul 19.
Artigo em Inglês | MEDLINE | ID: mdl-28682363

RESUMO

The motion of liquid metal has potential applications ranging from micro-pumps and self-fueled motors to rapid cooling and drug delivery. In this study, we systematically investigate the effects of the radius of LMDs (liquid metal droplets), the concentration of electrolyte solution and the applied electric field on the movement behavior of LMDs experimentally. The research also explains the experimental phenomenon with an innovative modeling analysis, which combines pertinent forces (i.e., the driving force induced by the gradient of surface tension, the viscous friction between the droplet and its surrounding electrolyte, and the friction between the droplet and the substrate). The model is highly consistent with the rule that LMDs with a larger radius need smaller actuation voltage, and we can predict the critical voltages of LMDs with r = 2-4 mm through Velectrode = 30.62/r2 - 0.998, which is obtained by fitting the parameters. We also obtain the model V = [-66.2Vr2/(259.7-17.7) + 1.253]r2, which can predict the average velocity-voltage lines of LMDs with r = 3, 3.5 mm and V = 1-13 V. In addition, the velocity increases upon increasing the concentration of the electrolyte solution from 0.1 mol L-1 to 0.3 mol L-1, and tends to be stable at more than 0.3 mol L-1 owing to the saturation of the EDL (electrical double layer) charge density. Additionally, we discuss the phenomenon of elongation during movement that occurs upon increasing the size of the LMDs. If the size of the LMDs continues to increase, the reverse movement from the anode to the cathode can occur, and the phenomenon can also be explained by the model.

12.
Animal ; 11(9): 1599-1607, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28077200

RESUMO

Omics research has indicated that heat shock protein 70 (HSP70) is a potential biomarker of meat quality. However, the specific changes and the potential role of HSP70 in postmortem meat quality development need to be further defined. In this study, Arbor Acres broiler chickens (n=126) were randomly categorized into three treatment groups of unstressed control (C), 0.5-h transport (T) and subsequent water shower spray following transport (T/W). Each treatment consisted of six replicates with seven birds each. The birds were transported according to a designed protocol. The pectoralis major (PM) muscles of the transport-stressed broilers were categorized as normal and pale, soft and exudative (PSE)-like muscle samples according to L* and pH24 h values to test the expression and location of HSP70. Results revealed that the activities of plasma creatine kinase and lactate dehydrogenase increased significantly (P<0.05) in normal and PSE-like muscle samples after transportation. The mRNA expression of HSP70 in normal muscle samples increased significantly (P<0.05) compared with that in the controls after stress. The protein expression of HSP70 increased significantly in normal muscle samples and decreased significantly (P<0.05) in PSE-like muscles. Immuno-fluorescence showed that HSP70 was present in the cytoplasm and on surface membranes of PM muscle cells in the normal samples following stress. Meanwhile, HSP70 was present on the surface membranes and extracellular matrix but was barely visible in the cytoplasm of the PSE-like samples. Principal component analysis showed high correlations between HSP70 and meat quality and stress indicators. In conclusion, this research suggests that the variation in HSP70 expression may provide a novel insight into the pathways underlying meat quality development.


Assuntos
Galinhas/fisiologia , Proteínas de Choque Térmico HSP70/genética , Carne/normas , Músculos Peitorais/fisiologia , Animais , Creatina Quinase/sangue , Proteínas de Choque Térmico HSP70/metabolismo , Masculino , Estresse Fisiológico , Transportes , Água/metabolismo
13.
Artigo em Inglês | MEDLINE | ID: mdl-27665746

RESUMO

BACKGROUND: It is gradually accepted that solid bolus swallow needs to be added to the procedure of manometry. The motility differences in the upper esophageal sphincter (UES) and lower esophageal sphincter (LES) were not well described. Sierra Scientific Instruments solid-state high-resolution manometry (HRM) system, the most popular HRM system in China, lacks the Chinese normative values for both liquid and solid bolus swallow parameters. METHODS: The esophageal HRM data of 88 healthy volunteers were analyzed. The parameters of both sphincters in resting stage were summarized and those during solid and liquid swallows were compared. KEY RESULTS: Normative HRM values of sphincter parameters in solid and liquid bolus swallows in China were established. The UES residual pressure of solid bolus swallows was lower than that of liquid bolus (0.3±5.5 mm Hg vs 4.8±5.9 mm Hg, P=.000). The time parameters of UES relaxation between two types of bolus swallows were similar. In solid bolus swallows, the intrabolus pressure (IBP) (13.8±5.1 mm Hg vs 10.9±5.7 mm Hg, P=.000) and LES relaxation time (11.0±2.1 seconds vs 8.7±1.3 seconds, P=.000) were higher. The 4-second integrated relaxation pressure between both bolus swallows was similar. CONCLUSIONS & INFERENCES: The function of the UES and LES between solid and liquid bolus swallows is different. Chinese HRM parameters are different from the Chicago Classification (http://www.chictr.org.cn, Number ChiCTR-EOC-15007147).


Assuntos
Deglutição/fisiologia , Ingestão de Líquidos/fisiologia , Ingestão de Alimentos/fisiologia , Esfíncter Esofágico Inferior/fisiologia , Motilidade Gastrointestinal/fisiologia , Manometria/métodos , Adulto , China/epidemiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
14.
Zhonghua Xue Ye Xue Za Zhi ; 37(11): 971-975, 2016 Nov 14.
Artigo em Chinês | MEDLINE | ID: mdl-27995883

RESUMO

Objective: To construct pIRES2-ZsGreen1/F Ⅸ expression vector, using the pcDNA/FⅨ plasmid containing FⅨ cDNA as template, and express in HEK-293 cells. Methods: The total ORF of F Ⅸ gene was amplified from pcDNA/F Ⅸ plasmid, then the amplified fragment was clonded into the pIRES2-ZsGreen1 vector using the Infusion enzyme. The positive clones of eukaryotic expression vector of pIRES2-ZsGreen1/F Ⅸ were screened and expanded after transfection, then were constructed and confirmed by PCR and sequencing. Transient expression experiments were performed using HEK-293 cells transfected with the expression vectors and observed the expression of ZsGreen1 protein by confocal laser microscope. The relative expression levels of FⅨ mRNA, protein and FⅨ activity (FⅨ∶C) were detected by real time PCR (RT-PCR), immunofluorescence microscopy, One-Stage method, respectively. Results: The expression vector, pIRES2-ZsGreen1/F Ⅸ, was successfully constructed and expressed in HEK-293 cells. RT-PCR detected the expression of F Ⅸ mRNA in HEK-293 cells and the immunofluorescence microscopy showed FⅨ protein distributed in the surrounding of nucleus. FⅨ∶C of cell lysates and cell culture fluid transfected with the expression vectors were (92.03 ± 0.29)% and (86.89 ± 8.78)%, respectively; while both F Ⅸ∶C of cell lysates and cell culture fluid transfected with or without the expression vectors were 0. Conclusion: The experimental results showed the expression vector, pIRES2-ZsGreen1/FⅨ, was successfully constructed , which provided experiment basement for the follow study on the location, function and molecular pathology of hemophilia B.


Assuntos
Fator IX/metabolismo , Expressão Gênica , Vetores Genéticos , Células HEK293 , Eucariotos , Proteínas de Fluorescência Verde , Humanos , Plasmídeos , RNA Mensageiro , Reação em Cadeia da Polimerase em Tempo Real , Proteínas Recombinantes , Transfecção
15.
Zhonghua Xue Ye Xue Za Zhi ; 37(5): 388-92, 2016 May 14.
Artigo em Chinês | MEDLINE | ID: mdl-27210873

RESUMO

OBJECTIVE: To construct human coagulation factor Ⅸ mini-gene (Mini-hF9) and some nonsense mutants, detect the levels of the Mini-hF9 mRNA, and analyze the molecular mechanism of microRNA125 regulating F9 gene with nonsense mutation. METHODS: Three nonsense mutants were obtained by using PCR mutagenesis to analyze the mechanism of plasma thromboplastin component recognition. The Mini-hF9 gene mRNA levels were detected by Real-time PCR in mammalian cells co-transfected with nonsense mutant expression vectors and miR-125 mimics. RESULTS: Mini-hF9 gene was constructed successfully and cloned into the mammalian expression vector. The only normal transcript was detected in cells transfected with the Mini-hF9 gene expression vectors. Three nonsense mutants, M1 (nt 34 G>T in Exon 7), M2 (nt 52 G>T in Exon 7) and M3 (nt 85 G>T in Exon 7), were obtained by using PCR mutagenesis. The levels of the Mini-hF9 mRNA decreased to 14.1% (t=15.464, P=0.004) in M1 and 22.4% (t=15.755, P=0.004) in M2 mutants after transfection, respectively. It was proved to be caused by nonsense-mediated mRNA decay (NMD) in CHX experiment. The levels of Mini-hF9 mRNA increased to 1.70 times (t=-4.883, P=0.039) and 2.40 times (t=-17.537, P=0.003) in M1 mutant after miR-125a or miR-125b mimics treatment, respectively. The levels of Mini-hF9 mRNA increased to 2.02 times (t=-19.264, P=0.003) and 2.07 times (t=-9.158, P=0.012) in M2 mutant after miR-125a or miR-125b mimics treatment, respectively. CONCLUSION: Nonsense mutant location is a key determinant for triggering NMD. MicroRNA125 could improve the stability of Mini-hF9 mRNA in M1 and M2 mutants by repressing NMD. MicroRNA125, a short non-coding RNA molecule, could be a potential therapeutic target in conditions caused by the NMD pathway.


Assuntos
Códon sem Sentido , Fator IX/genética , MicroRNAs/genética , Degradação do RNAm Mediada por Códon sem Sentido , Animais , Clonagem Molecular , Éxons , Expressão Gênica , Vetores Genéticos , Humanos , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Transfecção
16.
Mol Nutr Food Res ; 60(5): 1048-58, 2016 05.
Artigo em Inglês | MEDLINE | ID: mdl-26991948

RESUMO

SCOPE: Green tea has been known to confer numerous health benefits such as the prevention of cardiovascular disease, cancers, and obesity. Epigallocatechin-3-gallate (EGCG) is the major polyphenol present in green tea. Since EGCG is a food-derived component, intestinal epithelial cells lining the gastrointestinal tract are constantly and directly exposed to EGCG. It is anticipated that EGCG can exert beneficial effects in the intestine. The aim of this study was to explore the protective effects of EGCG on intestinal barrier functions against bacterial translocation by using a porcine jejunal epithelial cell line, IPEC-J2. METHODS AND RESULTS: EGCG reduced bacterial translocation across IPEC-J2 cell monolayers through the enhancement of the intestinal epithelial immunological barrier function by inducing secretion of antimicrobial peptides, porcine ß-defensins 1 and 2 (pBD-1 and 2), which possessed higher antimicrobial activity against Escherichia coli. Further mechanistic studies demonstrated that EGCG upregulated pBD-2 but not pBD-1 via the p38 mitogen-activated protein kinase dependent pathway. Such effects were not an "artifact" of hydrogen peroxide, catechin dimers, or other auto-oxidation products generated from EGCG in cell culture media. CONCLUSION: Our results imply that EGCG may be useful for prevention of intestinal disorders or bacterial infection in animals/humans.


Assuntos
Catequina/análogos & derivados , Intestinos/efeitos dos fármacos , Polifenóis/farmacologia , Chá/química , beta-Defensinas/metabolismo , Animais , Anti-Infecciosos/farmacologia , Catequina/farmacologia , Linhagem Celular , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Escherichia coli/efeitos dos fármacos , Mucosa Intestinal/metabolismo , Intestinos/citologia , Suínos , beta-Defensinas/farmacologia , Proteínas Quinases p38 Ativadas por Mitógeno/genética , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
17.
Br Poult Sci ; 56(1): 137-42, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25411077

RESUMO

1. The pharmacokinetics of doxycycline in ducks were investigated after a single intravenous (IV), intramuscular (IM) or oral (PO) dose at 20 mg/kg body weight. 2. The concentrations of doxycycline in plasma samples were assayed using a high performance liquid chromatography method, and pharmacokinetic parameters were calculated using a non-compartmental model. 3. After IV administration, doxycycline had a mean (±SD) distribution volume (Vz) of 1761.9 ± 328.5 ml/kg and was slowly eliminated with a terminal half-life (t1/2λz) of 21.21±1.47 h and a total body clearance (Cl) of 57.51 ± 9.50 ml/h/kg. Following PO and IM administration, doxycycline was relatively slowly absorbed - the peak concentrations (Cmax) were 17.57 ± 4.66 µg/ml at 2 h and 25.01 ± 4.18 µg/ml at 1.5 h, respectively. The absolute bioavailabilities (F) of doxycycline after PO and IM administration were 39.13% and 70.71%, respectively. 4. The plasma profile of doxycycline exhibited favourable pharmacokinetics characteristics in Muscovy ducks, such as wide distribution, relatively slow absorption and slow elimination, though oral bioavailability was low.


Assuntos
Doxiciclina/farmacocinética , Patos/metabolismo , Administração Oral , Animais , Antibacterianos/farmacocinética , Área Sob a Curva , Disponibilidade Biológica , Cromatografia Líquida de Alta Pressão/veterinária , Feminino , Meia-Vida , Injeções Intramusculares/veterinária , Injeções Intravenosas/veterinária , Masculino
18.
J Comp Pathol ; 149(2-3): 146-55, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23664424

RESUMO

Mast cells are key effectors in inflammation and contain proteinases that are released on activation. This study investigates associations between extracellular signal-regulated kinase (Erk)1/2, nuclear factor (NF)-κB, matrix metalloproteinase (MMP)-2 and MMP-9 in mast cells infected with Toxoplasma gondii tachyzoites. T. gondii infection led to increased mast cell degranulation. Phosphorylated (p)-Erk1/2 and p-NF-κB were increased significantly in mast cells infected with T. gondii. Pretreatment with the Erk kinase inhibitor PD98059 significantly decreased the expression of p-Erk1/2, p-NF-κB, MMP-2 and MMP-9. Treatment with MG132, an indirect NF-κB inhibitor, effectively reduced p-IκBα, p-NF-κB, MMP-2 and MMP-9 expression. Collectively, these data show that suppression of an Erk1/2/NF-κB signalling pathway caused a reduction in MMP-2 and -9 activities. Inhibiting this signalling pathway for MMP-2 and MMP-9 expression might offer a potential way to control early T. gondii infection. This pathway for the generation of MMP-2 and MMP-9 is important for mast cell secretion and the NF-κB/Erk1/2 signalling pathway may be key in MMP-2 and MMP-9 production in host defense against toxoplasmosis.


Assuntos
Mastócitos/metabolismo , Metaloproteinase 2 da Matriz/biossíntese , Metaloproteinase 9 da Matriz/biossíntese , Transdução de Sinais , Toxoplasmose/metabolismo , Animais , Western Blotting , Linhagem Celular , Sistema de Sinalização das MAP Quinases/imunologia , Mastócitos/imunologia , Mastócitos/microbiologia , Metaloproteinase 2 da Matriz/imunologia , Metaloproteinase 9 da Matriz/imunologia , Camundongos , NF-kappa B/imunologia , NF-kappa B/metabolismo , Transdução de Sinais/fisiologia , Toxoplasmose/imunologia , Regulação para Cima
19.
J Obstet Gynaecol ; 33(4): 370-4, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23654318

RESUMO

It has been suggested that in congenital Toxoplasma gondii infections, the parasite reaches the fetus by crossing the placental barrier. The purpose of this study was to determine the possible relationships between matrix metalloproteinases (MMPs) and dysfunction of the placental barrier in gravidas infected with T. gondii. We studied 26 umbilical cord sera; 20 and 6 were derived from gravidas seropositive for anti-T. gondii IgG and seropositive for anti-T. gondii IgM (low IgG avidity), respectively. Of 20 cord blood samples, 18 were seropositive for T. gondii IgG, whereas all cord blood samples were seronegative for T. gondii IgM. The other six sera were seronegative for T. gondii IgG, whereas three of six sera were seropositive for T. gondii IgM. Furthermore, T. gondii induced an increase in MMP-2 and -9 secretion in the sera of gravidas and umbilical cords. Moreover, MMP-2 and -9 were interacted with fibronectin. We propose that MMP-2 and -9 may be involved in ECM degradation and placental barrier dysfunction, which facilitates T. gondii transmission to the fetus. Future investigations of the effect of MMPs on migration across epithelial and endothelial barriers will be important to establish the mechanism of transit.


Assuntos
Fibronectinas/sangue , Metaloproteinase 2 da Matriz/sangue , Metaloproteinase 9 da Matriz/sangue , Toxoplasmose/sangue , Toxoplasmose/transmissão , Matriz Extracelular/metabolismo , Feminino , Sangue Fetal/química , Humanos , Placenta/fisiopatologia , Gravidez , Toxoplasmose/fisiopatologia , Toxoplasmose Congênita/etiologia
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