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2.
Biochem Biophys Res Commun ; 523(3): 580-587, 2020 Mar 12.
Artigo em Inglês | MEDLINE | ID: mdl-31941605

RESUMO

Sepsis induces critical myocardial dysfunction, resulting in an increased mortality. Gracillin (GRA) is a natural steroidal saponin, showing strong capacities of anti-inflammation, but its pharmacological effects on lipopolysaccharide (LPS)-induced acute cardiac injury still remain unclear. In this study, we attempted to explore if GRA was effective to attenuate cardiac injury in LPS-challenged mice and the underlying mechanisms. First, we found that GRA treatments markedly up-regulated the expression of miR-29a in cardiomyocytes. LPS-induced cytotoxicity in cardiomyocytes was significantly alleviated by GRA treatment, as evidenced by the improved cell viability and reduced lactate dehydrogenase (LDH) release. In addition, LPS-triggered apoptotic cell death was clearly ameliorated in cardiomyocytes co-treated with GRA. Notably, LPS-exposed cells showed significantly reduced expression of miR-29a, while being rescued by GRA treatment. In vivo, LPS apparently impaired cardiac function in mice, which was, however, alleviated by GRA administration. In addition, GRA markedly attenuated apoptosis in hearts of LPS-challenged mice by decreasing the expression of cleaved Caspase-3. LPS-triggered inflammatory response in cardiac tissues was also suppressed by GRA through blocking nuclear factor κB (NF-κB) signaling pathway. We also found that miR-29a expression was highly reduced in hearts of LPS-treated mice but was rescued by GRA pretreatment. Besides, miR-29a mimic alleviated LPS-induced apoptosis and inflammation in cardiomyocytes; however, LPS-caused effects were further accelerated by miR-29a. Of note, the protective effects of GRA on LPS-injured cardiac tissues were significantly abrogated by miR-29a suppression. In conclusion, our findings demonstrated that GRA exerted an effective role against LPS-induced acute cardiac injury through impeding apoptosis and inflammation regulated by miR-29a.

3.
Biotechnol Bioeng ; 117(2): 531-542, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31654413

RESUMO

Genetic manipulation is among the most important tools for synthetic biology; however, modifying multiple genes is extremely time-consuming and can sometimes be impossible when dealing with gene families. Here, we present a clustered regularly interspaced short palindromic repeat (CRISPR) and CRISPR-associated protein 9 (Cas9) system for use in the diploid yeast Candida tropicalis that is vastly superior to traditional techniques. This system enables the rapid and reliable introduction of multiple genetic deletions or mutations, as well as a stable expression using an integrated CRISPR-Cas9 cassette or a transient CRISPR-Cas9 cassette, together with a short donor DNA. We further show that the system can be used to promote the in vivo assembly of multiple DNA fragments and their stable integration into a target locus (or loci) in C. tropicalis. Based on this system, we present a platform for the biosynthesis of ß-carotene and its derivatives. These results enable the practical application of C. tropicalis and the application of the system to other organisms.

4.
Exp Ther Med ; 18(2): 1212-1220, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31316616

RESUMO

Long non-coding (lnc)RNA small nucleolar RNA host gene 12 (SNHG12) has an oncogenic role in various common human cancer types, including colorectal cancer (CRC). However, the detailed regulatory mechanisms of SNHG12 in CRC cells have remained largely elusive, and the investigation thereof was the purpose of the present study. Polymerase chain reaction analysis was performed to examine the expression of lncRNA and microRNA (miR). Cell Counting Kit-8 and Transwell assays were used to assess cell proliferation and invasion. A luciferase reporter assay was performed to confirm a predicted targeting association between lncRNA and miR. It was observed that SNHG12 was markedly upregulated in CRC tissues when compared with that in adjacent non-tumour tissues, and its high expression was associated with CRC progression, as well as poor prognosis of patients. In addition, the expression of SNHG12 was higher in CRC cell lines when compared with that in a normal intestinal epithelial cell line. Knockdown of SNHG12 significantly inhibited CRC cell proliferation and invasion, while ectopic overexpression of SNHG12 had the opposite effect. A Bioinformatics analysis predicted that SNHG12 and miR-16 have complementary binding sites, which was confirmed by a luciferase reporter gene assay. The expression levels of miR-16 were markedly decreased in CRC tissues and cell lines compared with those in normal tissues or cells, and were inversely correlated with the expression levels of SNHG12 in CRC tissues. Furthermore, silencing of miR-16 eliminated the suppressive effects of SNHG12 knockdown on CRC cell proliferation and invasion. In conclusion, the present study demonstrated that SNHG12 promotes CRC cell proliferation and invasion, at least in part, by acting as a molecular sponge of miR-16, suggesting that SNHG12 may be a promising therapeutic target for CRC.

5.
Oncol Res ; 26(4): 593-604, 2018 May 07.
Artigo em Inglês | MEDLINE | ID: mdl-28470146

RESUMO

MicroRNAs (miRs), a class of small noncoding RNAs, are important regulators for gene expression through directly binding to the 3'-untranslated region (3'-UTR) of their target mRNA. Recently, downregulation of miR-520b has been observed in several common human cancers. However, the exact role of miR-520b in colorectal cancer (CRC) has not previously been studied. In this study, our data showed that miR-520b was significantly downregulated in CRC and cell lines when compared with adjacent normal tissues and a normal intestinal epithelial cell line. Low expression of miR-520b was notably associated with the malignant progress and a shorter survival time for CRC patients. Restoration of miR-520b inhibited cell proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) in CRC cells. Defective in cullin neddylation 1 domain containing 1 (DCUN1D1) was then identified as a novel target gene of miR-520b in CRC cells. The expression of DCUN1D1 was significantly increased in CRC, with a negative correlation to miR-520b expression in CRC tissues. Moreover, a high expression of DCUN1D1 was significantly associated with the malignant progress and a poor prognosis for CRC patients. Furthermore, overexpression of DCUN1D1 rescued the miR-520b-mediated malignant phenotypes and EMT in CRC cells. The data demonstrate that miR-520b functions as a tumor suppressor in CRC through targeting DCUN1D1, suggesting that miR-520b may become a potential therapeutic target for the treatment of CRC.


Assuntos
Neoplasias Colorretais/genética , Genes Supressores de Tumor , MicroRNAs/genética , Proteínas Proto-Oncogênicas/antagonistas & inibidores , Regiões 3' não Traduzidas/genética , Idoso , Neoplasias Colorretais/mortalidade , Neoplasias Colorretais/patologia , Feminino , Células HCT116 , Células HT29 , Humanos , Masculino , Pessoa de Meia-Idade , Proteínas , Proteínas Proto-Oncogênicas/genética , Análise de Sobrevida
6.
Mol Cell Biochem ; 431(1-2): 123-131, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28281187

RESUMO

An increasing body of evidence indicates that inflammation and apoptosis are involved in the development of acute myocardial infarction (AMI). In this study, we sought to investigate the specific role and the underlying regulatory mechanism of miR-145-5p in myocardial ischemic injury. H9c2 cardiac cells were exposed to hypoxia to establish a model of myocardial hypoxic/ischemic injury. We found that miR-145-5p was notably down-regulated, while CD40 expression was highly elevated in H9c2 cells following exposure to acute hypoxia. Additionally, hypoxia markedly enhanced the inflammatory response, as reflected by an increase in the secretion of the cytokines IL-1ß, TNF-α, and IL-6, whereas the introduction of miR-145-5p effectively suppressed inflammatory factor production triggered by hypoxia. Furthermore, we observed hypoxia stimulation significantly augmented apoptosis accompanied by a decrease in the expression of Bcl-2 and an increase in the expression of Bax, Caspase-3, and Caspase-9. However, augmentation of miR-145-5p led to a dramatic prevention of hypoxia-induced apoptosis. Importantly, we identified CD40 as a direct target of miR-145-5p. Interestingly, the depletion of CD40 with small interfering RNAs (siRNAs) apparently repressed the production of inflammatory cytokines and apoptosis in the setting of acute hypoxic treated. Taken together, these data demonstrated that miR-145-5p may function as a cardiac-protective molecule in myocardial ischemic injury by ameliorating inflammation and apoptosis via negative regulation of CD40. The study gives evidence that miR-145-5p provides an interesting strategy for protecting cardiomyocytes from hypoxia-induced inflammatory response and apoptosis.


Assuntos
Apoptose , Antígenos CD40/metabolismo , MicroRNAs/metabolismo , Infarto do Miocárdio/metabolismo , Miócitos Cardíacos/metabolismo , Proteínas Reguladoras de Apoptose/metabolismo , Hipóxia Celular , Linhagem Celular , Citocinas/metabolismo , Humanos , Inflamação/metabolismo , Inflamação/patologia , Infarto do Miocárdio/patologia , Miócitos Cardíacos/patologia
7.
Sci Rep ; 6: 25874, 2016 05 13.
Artigo em Inglês | MEDLINE | ID: mdl-27174789

RESUMO

We seek to confirm the effect and explore the indications of aggressive locoregional management in patients with metastatic inflammatory breast cancer (IBC). Between 2003 and 2014, we reviewed the records of 156 patients with metastatic IBC from five large centers of Breast Surgery in the region of central south of China. Clinicopathologic data were collected to access overall survival (OS), prognostic factors and the indications for locoregional treatment. 75 (48%) patients underwent aggressive locoregional therapy. Patients in locoregional therapy group had a median OS of 24 months compared with 17 months of those in no locoregional therapy group. 2-year OS rate of these two groups was 52% and 32%, separately. Locoregional therapy (HR = 0.556; 95% CI 0.385-0.803; p = 0.002) was confirmed to be an independent prognostic factor, which could significantly improve OS of patients with metastatic IBC. For locoregional therapy group, statistical differences were observed in all subgroups stratified by the factors that were significant in univariate analysis except in the subgroups of stable disease, Charlson comorbidity index ≥3 and cerebral metastasis. Therefore, systemic therapy efficacy, Charlson comorbidity index and cerebral metastasis status appeared to be important indexes for choice of locoregional therapy in different individuals.


Assuntos
Terapia Combinada/métodos , Neoplasias Inflamatórias Mamárias/patologia , Neoplasias Inflamatórias Mamárias/terapia , China , Feminino , Humanos , Gradação de Tumores , Metástase Neoplásica , Seleção de Pacientes , Medicina de Precisão , Prognóstico , Análise de Sobrevida , Resultado do Tratamento
8.
J Cancer Res Clin Oncol ; 141(2): 345-52, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25178995

RESUMO

OBJECTIVES: We seek to summarize the clinicopathologic characteristics and prognostic factors of sarcomatoid renal cell carcinoma (SRCC), an uncommon type of renal cell carcinoma. METHODS AND MATERIALS: Between 2004 and 2012, 23 patients with SRCC were treated at a large urology center in south central China. We collect patient's clinicopathologic features from medical records to assess diagnosis, prognostic factors and efficacy of systemic therapy. Clinical data were absent in 3 cases, and 20 patients were enrolled in the final study. RESULTS: Immunohistochemically, almost all SRCC expressed cytokeratin (91%), epithelial membrane antigen (87%) and vimentin (100%). Sarcomatoid differentiation occurs in various kinds of subtypes of RCC with almost the same probability. The median tumor size was 10.5 cm. The CT findings of these tumors revealed low-density (n = 5; 25%) or mixed (n = 15; 75%) masses with necrotic areas and often showed an infiltrative morphology (n = 15; 75%). All 20 cases demonstrated heterogeneous enhancement, and eleven (55%) cases demonstrated >50% necrosis. Six cases complicated with calculus and hydronephrosis. Sixteen (80%) patients demonstrated invasions of tissues localized in Gerota's fascia, and 8 (40%) tumors invaded beyond Gerota's fascia. Fifteen (75%) patients demonstrated lymph node metastasis, and sixteen (80%) patients had distant metastasis. Five patients received systemic therapy, and one patient given high-dose interferon-α had a completely response, and one patient received chemotherapy based on gemcitabine had partial response. The median overall survival of all patients was 5.8 months. Patients without distant metastasis had a median overall survival of 35 months compared with 3 months of those with distant metastasis (P < 0.002). The percentage of the sarcomatoid components did not have an obvious influence to the prognosis (P = 0.197). CONCLUSIONS: Heterogeneity, hugeness, infiltration and necrosis are typical image features of SRCC. The prognosis of SRCC is poor and clinic stage especially the existence of distant disease is the important factor influencing prognosis.


Assuntos
Carcinoma de Células Renais/secundário , Neoplasias Renais/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma de Células Renais/mortalidade , Carcinoma de Células Renais/terapia , Terapia Combinada , Feminino , Seguimentos , Humanos , Neoplasias Renais/mortalidade , Neoplasias Renais/terapia , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica , Estadiamento de Neoplasias , Prognóstico , Taxa de Sobrevida
9.
Saudi Med J ; 35(11): 1324-30, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25399208

RESUMO

OBJECTIVES: To determine the outcome of patients with luminal A, luminal B, human epidermal growth factor receptor-2 (HER-2) positive, and triple negative molecular subtypes of inflammatory breast cancer (IBC) using a retrospective analysis. METHODS: This study was conducted between February 2004 and February 2010 in 3 different hospitals in China. The clinical outcomes, pathological features, and treatment strategies were analyzed in 67 cases of IBC without distant metastases. A chi-square test and one-way ANOVA were used to assess outcomes between different subtypes. Overall survival (OS) was analyzed using the Kaplan-Meier method and multivariate analysis was conducted using the Cox regression model. RESULTS: The 2-year OS rate was 55% for the entire cohort. Median OS time among patients with luminal A was 35 months, luminal B was 30 months, HER-2 positive was 24 months, and triple negative subtypes was 20 months, and were significantly different from each other (p=0.001). Using multivariate analysis, luminal A had 76% (p=0.037), luminal B had 54% (p=0.048), and HER-2 positive subtypes had 47% (p=0.032) decreased risk of death compared with the triple negative subtype.  Furthermore, elevated Ki-67 labeling was associated with increased risk of death, while the surgical treatment significantly improved patient survival. CONCLUSION: Breast cancer subtypes are associated with distinct outcomes in IBC patients. Patients that presented with triple negative IBC had poorer outcome than luminal A, luminal B, and HER-2 subtypes. These results indicate that IBC is a heterogeneous disease similar to the conventional breast cancer. 


Assuntos
Antineoplásicos/uso terapêutico , Carcinoma/metabolismo , Neoplasias Inflamatórias Mamárias/metabolismo , Antígeno Ki-67/metabolismo , Receptor ErbB-2/metabolismo , Receptores Estrogênicos/metabolismo , Receptores de Progesterona/metabolismo , Neoplasias de Mama Triplo Negativas/metabolismo , Adulto , Idoso , Carcinoma/classificação , Carcinoma/terapia , Feminino , Humanos , Neoplasias Inflamatórias Mamárias/classificação , Neoplasias Inflamatórias Mamárias/terapia , Mastectomia , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Prognóstico , Modelos de Riscos Proporcionais , Neoplasias de Mama Triplo Negativas/classificação , Neoplasias de Mama Triplo Negativas/terapia
10.
ScientificWorldJournal ; 2014: 530942, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25147853

RESUMO

Few researches pay attention to prediction about analog circuits. The few methods lack the correlation with circuit analysis during extracting and calculating features so that FI (fault indicator) calculation often lack rationality, thus affecting prognostic performance. To solve the above problem, this paper proposes a novel prediction method about single components of analog circuits based on complex field modeling. Aiming at the feature that faults of single components hold the largest number in analog circuits, the method starts with circuit structure, analyzes transfer function of circuits, and implements complex field modeling. Then, by an established parameter scanning model related to complex field, it analyzes the relationship between parameter variation and degeneration of single components in the model in order to obtain a more reasonable FI feature set via calculation. According to the obtained FI feature set, it establishes a novel model about degeneration trend of analog circuits' single components. At last, it uses particle filter (PF) to update parameters for the model and predicts remaining useful performance (RUP) of analog circuits' single components. Since calculation about the FI feature set is more reasonable, accuracy of prediction is improved to some extent. Finally, the foregoing conclusions are verified by experiments.


Assuntos
Modelos Teóricos , Algoritmos
11.
Virology ; 464-465: 166-176, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25083619

RESUMO

The H9N2 avian influenza virus (AIV) has become increasingly concerning due to its role in severe economic losses in the poultry industry. Transmission of AIV to mammals, including pigs and humans, has accelerated efforts to devise preventive strategies. To develop an effective oral vaccine against H9N2 AIV, a recombinant Lactobacillus plantarum NC8 strain expressing the hemagglutinin (HA) gene of H9N2 AIV was constructed in this study. Mice were orally immunized with the recombinant NC8-pSIP409-HA strain, and sIgA, IgG and HI antibodies were produced by the NC8-pSIP409-HA strain, which also induced CD8(+) T cell immune responses. Most importantly, oral administration produced complete protection against challenge with mouse-adapted H9N2 virus. These results indicate that the recombinant NC8-pSIP409-HA was more effective at inducing the mucosal, humoral and cellular immune responses. Therefore, L. plantarum NC8-pSIP409-HA could become a promising oral vaccine candidate against H9N2 AIV.


Assuntos
Glicoproteínas de Hemaglutininação de Vírus da Influenza/imunologia , Vírus da Influenza A Subtipo H9N2/imunologia , Vacinas contra Influenza/imunologia , Influenza Aviária/prevenção & controle , Influenza Humana/prevenção & controle , Doenças das Aves Domésticas/prevenção & controle , Animais , Anticorpos Antivirais/imunologia , Galinhas , Feminino , Expressão Gênica , Glicoproteínas de Hemaglutininação de Vírus da Influenza/administração & dosagem , Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Humanos , Vírus da Influenza A Subtipo H9N2/genética , Vacinas contra Influenza/administração & dosagem , Vacinas contra Influenza/genética , Influenza Aviária/imunologia , Influenza Aviária/virologia , Influenza Humana/imunologia , Influenza Humana/virologia , Lactobacillus/genética , Lactobacillus/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Doenças das Aves Domésticas/imunologia , Doenças das Aves Domésticas/virologia , Vacinação
12.
Comput Intell Neurosci ; 2014: 740838, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25610458

RESUMO

This paper proposes a novel test generation algorithm based on extreme learning machine (ELM), and such algorithm is cost-effective and low-risk for analog device under test (DUT). This method uses test patterns derived from the test generation algorithm to stimulate DUT, and then samples output responses of the DUT for fault classification and detection. The novel ELM-based test generation algorithm proposed in this paper contains mainly three aspects of innovation. Firstly, this algorithm saves time efficiently by classifying response space with ELM. Secondly, this algorithm can avoid reduced test precision efficiently in case of reduction of the number of impulse-response samples. Thirdly, a new process of test signal generator and a test structure in test generation algorithm are presented, and both of them are very simple. Finally, the abovementioned improvement and functioning are confirmed in experiments.


Assuntos
Algoritmos , Inteligência Artificial , Reconhecimento Automatizado de Padrão , Processamento de Sinais Assistido por Computador , Humanos , Modelos Teóricos , Fatores de Tempo
13.
Basic Res Cardiol ; 108(3): 341, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23463182

RESUMO

The N-myc downstream-regulated gene 2 (NDRG2) is involved in cell apoptosis and survival. Although reported to be highly expressed in the cardiac tissue, the biological function of NDRG2 in the heart remains to be established. Insulin exerts protective effects against myocardial ischemia/reperfusion (I/R) injury through the PI3K/Akt pathway. Here, we examined the changes in phosphorylation of NDRG2, a novel substrate and phosphoprotein of Akt, in insulin-induced protection against myocardial I/R. Rat hearts were subjected to 30 min regional ischemia followed by reperfusion with or without insulin at the onset of reperfusion. Reperfusion with insulin inhibited myocardial apoptosis and reduced infarct size, as well as significantly up-regulated myocardial Akt and NDRG2 phosphorylation levels compared with the I/R group. These effects of insulin were blocked by pretreatment with the PI3K inhibitor wortmannin or Akt inhibitor. To further ascertain the role of NDRG2 in insulin-induced cardioprotection, cardiomyocytes were transduced with a lentivirus encoding shRNA targeting NDRG2 (loss-of-function), which rendered the cells more susceptible to I/R injury and significantly blunted the anti-apoptotic effect of insulin. Moreover, the NDRG2 shRNA lentivirus was tested in vivo, and NDRG2 knockdown aggravated myocardial I/R injury and attenuated the insulin-mediated cardioprotection against I/R injury. Taken together, these results suggest a novel role of PI3K/Akt/NDRG2 signaling in the cardioprotective effect of insulin.


Assuntos
Cardiotônicos/farmacologia , Insulina/farmacologia , Infarto do Miocárdio/prevenção & controle , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Miocárdio/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Função Ventricular Esquerda/efeitos dos fármacos , Animais , Animais Recém-Nascidos , Apoptose/efeitos dos fármacos , Células Cultivadas , Modelos Animais de Doenças , Masculino , Contração Miocárdica/efeitos dos fármacos , Infarto do Miocárdio/metabolismo , Infarto do Miocárdio/patologia , Infarto do Miocárdio/fisiopatologia , Traumatismo por Reperfusão Miocárdica/metabolismo , Traumatismo por Reperfusão Miocárdica/patologia , Traumatismo por Reperfusão Miocárdica/fisiopatologia , Miocárdio/patologia , Proteínas do Tecido Nervoso/genética , Fosfatidilinositol 3-Quinase/metabolismo , Fosforilação , Inibidores de Proteínas Quinases/farmacologia , Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-akt/metabolismo , Interferência de RNA , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos , Volume Sistólico/efeitos dos fármacos , Fatores de Tempo , Transfecção
14.
J Asian Nat Prod Res ; 13(6): 512-22, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21623514

RESUMO

Respiratory syncytial virus (RSV) is the most important cause of lower respiratory tract infection in infants and young children. With the emergence of drug-resistant strains of RSV, new antiviral agents are needed urgently. Gentiana rigescens is a kind of Chinese herb, belonging to Gentianaceae, which has long been used as a folk medicine for curing inflammation, bacterial infection, viral infection, and so on. In this research, polysaccharide designated RG4-1 was isolated from G. rigescens by hot water extraction, ethanol precipitation, and macroreticular adsorbing resin column chromatography, and its antiviral activity, cytotoxicity, and possible antiviral mechanisms were assayed by cytopathogenic effect inhibition assay, 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, and plaque reduction assay. RG4-1 was a fructose-binding lectin. In host cell cultures, RG4-1 was found to be an effective antiviral component against RSV. It showed good inhibitory effect against RSV when it was added 2 h after virus infection with 50% effective concentration of 12.86 µg/ml. RG4-1 also displayed its direct inactivation, attachment inhibition effect, and penetration inhibition effect against RSV. A time-dependent experiment was set up to confirm that RG4-1 blocked RSV infection at early stages of the infection. But RG4-1 seemed to be ineffective against intracellular virus and viral biosynthesis.


Assuntos
Antivirais/isolamento & purificação , Antivirais/farmacologia , Gentiana/química , Polissacarídeos/isolamento & purificação , Polissacarídeos/farmacologia , Infecções por Vírus Respiratório Sincicial/tratamento farmacológico , Vírus Sinciciais Respiratórios/efeitos dos fármacos , Replicação Viral/efeitos dos fármacos , Animais , Criança , Relação Dose-Resposta a Droga , Humanos , Lactente , Sais de Tetrazólio/farmacocinética , Tiazóis/farmacocinética , Fatores de Tempo , Células Vero
15.
Peptides ; 31(2): 271-4, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-20004694

RESUMO

Visfatin is a 56 kDa protein that is overexpressed in pregnant women. Like insulin, 2 nM visfatin induced GLUT 4 translocation from the cytosolic fraction to the membrane in 3T3-L1 cells. We have previously reported that visfatin induces glucose uptake into 3T3-L1 cells. These two actions define visfatin as an insulinomimetic. Three estrogens are elevated in pregnancy. Estradiol, the predominant estrogen, estriol, produced by the fetal liver and the pro-estrogen progesterone are all higher during pregnancy than in nonparous women. 3T3-L1 cells were treated with 150 ng/ml estriol, 16 ng/ml estradiol or 190 ng/ml progesterone to reflect the circulating concentrations of these steroids during pregnancy. Estriol treatment produced a 2.5-fold increase in visfatin gene expression. Estradiol and progesterone had small but insignificant effects on visfatin gene expression. In a second experiment, cells were treated with a combination of all three steroids together at the same concentrations listed above. The combination treatment produced a 13-fold increase in visfatin gene expression. These data suggest that the estriol, estradiol and progesterone exert a synergistic effect on visfatin gene expression. Taken together these data suggest that visfatin may play a physiological role during pregnancy. Since visfatin potently and efficaciously induced GLUT 4 translocation in a cell culture model, any hypothetical role for visfatin in pregnancy should include the possibility that it may play a role in maternal/fetal glucose metabolism or distribution. Two possibilities present: either maternal visfatin is overexpressed as a protective response in the pregnant female to compensate for the insulin resistance that often accompanies pregnancy or the excess visfatin is a compensatory response to ensure adequate glucose delivery to the growing fetus.


Assuntos
Citocinas/genética , Estrogênios/farmacologia , Expressão Gênica/genética , Nicotinamida Fosforribosiltransferase/genética , Células 3T3-L1 , Animais , Membrana Celular/metabolismo , Citocinas/farmacologia , Citoplasma/metabolismo , Estradiol/farmacologia , Estriol/farmacologia , Feminino , Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/fisiologia , Transportador de Glucose Tipo 4/metabolismo , Insulina/farmacologia , Camundongos , Nicotinamida Fosforribosiltransferase/farmacologia , Gravidez , Progesterona/farmacologia
16.
Wei Sheng Yan Jiu ; 38(6): 733-5, 2009 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-20047236

RESUMO

OBJECTIVE: To study the extraction of an active component LC-4 from Forsythia suspensa (Thunb.) Vahl and anti-respiratory syncytial virus (RSV) effect in vitro. METHODS: The extration from boiling water of Forsythia suspensa were extracted by hot water extraction and were precipitated by ethanol and were elicited by macroporous adsorptive resins column chromatography. Then the antiviral active component LC-4 was separated from the production by eluting. The cytotoxicities and antiviral effect of LC-4 were tested by the Neutral Red assay and the cytopathic effect (CPE) inhibition assay. Ribavirin was used as the positive drug in the study. The antiviral effect of prevention and treatment of maximum dose of non-cytotoxicity of LC-4 was tested in Hela cell line. RESULTS: LC-4 showed that the concentration were 25.4 microg/ml with the medium cytotoxicity and were 2.11 microg/ml with the medium effective concentration (EC50). The corresponding treatment index of LC-4 was 12.04. It also exhibited obviously inhibition effect on RSV replications when LC-4 was added at 2 h, 4 h, 6 h and 8 h after infection (P < 0.05). The CPE was significant difference in comparison with control group, when the concentration of LC-4 between 31.25 microg/ml and 3.91 microg/ml was added at 2 h before infection (P < 0.05). CONCLUSION: LC-4 could significantly prevent and inhibit RSV replications in vitro.


Assuntos
Antivirais/farmacologia , Medicamentos de Ervas Chinesas/farmacologia , Forsythia/química , Vírus Sinciciais Respiratórios/efeitos dos fármacos , Antivirais/isolamento & purificação , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/isolamento & purificação , Células HeLa , Humanos
17.
J Virol Methods ; 153(2): 218-22, 2008 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-18652846

RESUMO

Influenza is a serious global public health problem and an economic burden. With the continual emergence of new influenza A virus strains, new antiviral drugs are needed urgently. In this study, an improved embryonated chicken egg model for evaluating antiviral activity against Influenza A virus was developed. In the model, the influenza A virus was injected into the allantoic cavity and ribavirin was injected into the albumen of the egg. The levels of influenza A virus in the allantoic fluid was titrated by the hemagglutination test after incubation for 72 h at 35.5 degrees C and 12 h at 4 degrees C. Ribavirin treatment at a dose of 25 mg/kg to 100 mg/kg decreased significantly the hemagglutination titers both of Influenza virus A/FM1, H1N1 (IVA1) (p < 0.01) and influenza virus A/Wuhan/359/95, H3N2 (IVA3) (p < 0.01). In a time-dependent drug addition assay, significant efficacy of ribavirin against both IVA1 and IVA3 was observed when the drug was administered before and shortly after viral inoculation (p < 0.01 or p < 0.05). In conclusion, ribavirin treatment showed significant antiviral activity against IVA1 and IVA3 in this model, suggesting that the improved model would be useful for evaluating the anti-influenza virus activity of potential inhibitors.


Assuntos
Alantoide/virologia , Antivirais/farmacologia , Vírus da Influenza A Subtipo H1N1/efeitos dos fármacos , Vírus da Influenza A Subtipo H3N2/efeitos dos fármacos , Testes de Sensibilidade Microbiana/métodos , Ribavirina/farmacologia , Animais , Antivirais/administração & dosagem , Linhagem Celular , Embrião de Galinha , Testes de Hemaglutinação , Humanos , Modelos Animais , Ribavirina/administração & dosagem
18.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 13(6): 1106-8, 2005 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-16403291

RESUMO

In order to investigate the effect of antioxidants on human blood, vitamin C was selected and added into plastic blood storage bags with CPD, and stored at 25 degrees C. During 6 days of storage, some indexes as ATP, SOD, MDA, K(+) concentration and superoxide radicals were detected and were compared with control group, The results showed that ATP and SOD activity in whole blood with vitamin C during 6 days of storage was higher then that in control group (P < 0.05, P < 0.01), the MDA and plasma K(+) concentrations in stored whole blood with vitamin C during 6 days of storage were lower than that in control group (P < 0.05, P < 0.01), the superoxide radical concentrations in stored whole blood with vitamin C decreased lower than that in control group (30%). The conclusion was made that vitamin C increases activities of ATP and SOD, decreases concentrations of MDA, plasma K(+) and superoxide radicals during blood preservation.


Assuntos
Antioxidantes/farmacologia , Ácido Ascórbico/farmacologia , Preservação de Sangue/métodos , Eritrócitos/efeitos dos fármacos , Trifosfato de Adenosina/sangue , Eritrócitos/citologia , Eritrócitos/metabolismo , Humanos , Malondialdeído/sangue , Potássio/sangue , Superóxido Dismutase/sangue , Fatores de Tempo
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