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1.
Chin Med J (Engl) ; 2020 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-32149769

RESUMO

BACKGROUND: Medicines for the treatment of 2019-novel coronavirus (2019-nCoV) infections are urgently needed. However, drug screening using live 2019-nCoV requires high-level biosafety facilities, which imposes an obstacle for those without such facilities or 2019-novel coronavirus (2019-nCoV). This study aims to repurpose the clinically approved drugs for the treatment of coronavirus disease 2019 (COVID-19) in a 2019-nCoV related coronavirus model. METHODS: A 2019-nCoV related pangolin coronavirus GX_P2V/pangolin/2017/ Guangxi was described. Whether GX_P2X uses angiotensin-converting enzyme 2 (ACE2) as the cell receptor was investigated by using small interfering RNA (siRNA) -mediated silencing of ACE2. The pangolin coronavirus model was used to identify drug candidates for treating 2019-nCoV infection. Two libraries of 2406 clinically approved drugs were screened for their ability to inhibit cytopathic effects on Vero E6 cells by GX_P2X infection. The antiviral activities and antiviral mechanisms of potential drugs were further investigated. Viral yields of RNAs and infectious particles were quantified by quantitative real-time polymerase chain reaction (qRT-PCR) and plaque assay, respectively. RESULTS: The spike protein of coronavirus GX_P2V shares 92.2% amino acid identity with that of 2019-nCoV isolate Wuhan-hu-1, and uses ACE2 as the receptor for infection just like 2019-nCoV. Three drugs-cepharanthine (CEP), selamectin and mefloquine hydrochloride exhibited complete inhibition of cytopathic effects in cell culture at 10 µmol/L. CEP demonstrated the most potent inhibition of GX_P2V infection, with a concentration for 50% of maximal effect [EC50] of 0.98 µmol/L. The viral RNA yield in cells treated with 10 µmol/L CEP was 15,393-fold lower than in cells without CEP treatment ([6.48 ±â€Š0.02] × 10vs. 1.00 ±â€Š0.12, t = 150.38, P < 0.001) at 72 h post-infection (p.i.). Plaque assays found no production of live viruses in media containing 10 µmol/L CEP at 48 h p.i. Furthermore, we found CEP has potent antiviral activities against both viral entry (1.00 ±â€Š0.37 vs. 0.46 ±â€Š0.12, t = 2.42, P < 0.05) and viral replication (1.00 ±â€Š0.43 vs. [6.18 ±â€Š0.95] × 10, t = 3.98, P < 0.05). CONCLUSIONS: Our pangolin coronavirus GX_P2V is a workable model for 2019-nCoV research. CEP, selamectin and mefloquine hydrochloride are potential drugs for treating 2019-nCoV infection. Our results strongly suggest that CEP is a wide-spectrum inhibitor of pan-betacoronavirus, and clinical trial of CEP for treatment of 2019-nCoV infection is warranted.

2.
Horm Cancer ; 10(4-6): 177-189, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31713780

RESUMO

In hepatocellular carcinoma (HCC), the hypoxic tumor microenvironment can drive enhance tumor malignancy and recurrence. The microRNA (miRNA) miR-196-5p has been shown to modulate the progression of several cancer types, but its roles in HCC remain uncertain. In the present report we observed significant miR-196-5p downregulation in HCC tissues and cells, and we found that the expression of this miRNA significantly impaired the proliferation and metastatic potential of HCC in vitro and in vivo. We identified high-mobility group AT-hook 2 (HMGA2) as a miR-196-5p target gene that was associated with the ability of miR-196-5p to modulate the progression of HCC. Expression of miR-196-5p and HMGA2 were correlated with the clinical characteristics and poor outcomes in patients with HCC. Finally, we found that hypoxic conditions were linked with reduced miR-196-5p expression in the context of HCC. Together these results highlight the role for miR-196-5p as an inhibitor of the proliferation and metastasis of HCC via the targeting of HMGA2, with this novel hypoxia/miR-196-5p/HMGA2 pathway serving as a potential target for future therapeutic intervention.

3.
Food Chem ; 299: 125095, 2019 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-31279124

RESUMO

Hydrogen gas (H2), a multifunctional signaling molecule, has received increasing attention in recent years. In the present study, hydrogen-rich water (HRW) (2 ppm) was used for the processing of sprouted black barley (Hordeum distichum L.), and the results showed that the HRW treatment could significantly increase the germination rate and growth rate of black barley (P < 0.05). A chemical component analysis showed that in sprouted black barley, the HRW treatment could change the distribution of phytochemicals (e.g., the ionic strength of guanosine), increase the concentrations of free vanillic acid, coumaric acid, sinapic acid, conjugated sinapic acid, Ca and Fe and the hydroxyl radical scavenging rate, and decrease the protein, fat, starch and dietary fibre contents compared with the results obtained after treatment with ultra-pure water (P < 0.05). HRW can be used for the processing of sprouted grains to effectively increase their germination efficiency and concentrations of bioactive phytochemicals.


Assuntos
Hordeum/química , Hordeum/crescimento & desenvolvimento , Hidrogênio/farmacologia , Antioxidantes/análise , Cromatografia Líquida , Fibras na Dieta/análise , Germinação/efeitos dos fármacos , Hordeum/efeitos dos fármacos , Radical Hidroxila/análise , Espectrometria de Massas , Proteínas de Vegetais Comestíveis/análise , Amido/análise , Água/química , Água/farmacologia
4.
Biochem Pharmacol ; 150: 191-201, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29454618

RESUMO

Bigelovin, a sesquiterpene lactone, has been demonstrated to induce apoptosis, inhibit inflammation and angiogenesis in vitro, but its potential anti-metastatic activity remains unclear. In the present study, two colon cancer mouse models, orthotopic tumor allografts and experimental metastatic models were utilized to investigate the progression and metastatic spread of colorectal cancer after bigelovin treatments. Results showed that bigelovin (intravenous injection; 0.3-3 mg/kg) significantly suppressed tumor growth and inhibited liver/lung metastasis with modulation of tumor microenvironment (e.g. increased populations of T lymphocytes and macrophages) in orthotopic colon tumor allograft-bearing mice. Furthermore, the inhibitory activities were also validated in the experimental human colon cancer metastatic mouse model. The underlying mechanisms involved in the anti-metastatic effects of bigelovin were then revealed in murine colon tumor cells colon 26-M01 and human colon cancer cells HCT116. Results showed that bigelovin induced cytotoxicity, inhibition of cell proliferation, motility and migration in both cell lines, which were through interfering IL6/STAT3 and cofilin pathways. Alternations of the key molecules including Rock, FAK, RhoA, Rac1/2/3 and N-cadherin, which were detected in bigelovin-treated cancer cells, were also observed in the tumor allografts of bigelovin-treated mice. These findings strongly indicated that bigelovin has potential to be developed as anti-tumor and anti-metastatic agent for colorectal cancer.


Assuntos
Neoplasias Colorretais/tratamento farmacológico , Inibidores do Crescimento/administração & dosagem , Interleucina-6/metabolismo , Lactonas/administração & dosagem , Fator de Transcrição STAT3/metabolismo , Sesquiterpenos/administração & dosagem , Transdução de Sinais/efeitos dos fármacos , Animais , Movimento Celular/efeitos dos fármacos , Movimento Celular/fisiologia , Neoplasias Colorretais/metabolismo , Células HCT116 , Humanos , Injeções Intravenosas , Interleucina-6/antagonistas & inibidores , Masculino , Camundongos , Camundongos Nus , Fator de Transcrição STAT3/antagonistas & inibidores , Transdução de Sinais/fisiologia , Ensaios Antitumorais Modelo de Xenoenxerto/métodos
5.
Sci Rep ; 7: 42176, 2017 02 09.
Artigo em Inglês | MEDLINE | ID: mdl-28181527

RESUMO

Colorectal cancer (CRC) is the third most prevalent cancer and the third highest cancer-related mortality in the United States. Bigelovin, a sesquiterpene lactone isolated from Inula helianthus aquatica, has been proven to induce apoptosis and exhibit anti-inflammatory and anti-angiogenic activities. However, the effects of bigelovin on CRC and underlying mechanisms have not been explored. The present study demonstrated that bigelovin exhibited potent anti-tumor activities against CRC in vitro and in vivo. Bigelovin suppressed cell proliferation and colony formation and induced apoptosis in human colorectal cancer HT-29 and HCT 116 cells in vitro. Results also revealed that bigelovin activated caspases, caused the G2/M cell cycle arrest and induced DNA damage through up-regulation of death receptor (DR) 5 and increase of ROS. In HCT 116 xenograft model, bigelovin treatment resulted in suppression of tumor growth. Bigelovin at 20 mg/kg showed more significant tumor suppression and less side effects than conventional FOLFOX (containing folinic acid, 5-fluorouracil and oxaliplatin) treatment. In addition, in vivo data confirmed that anti-tumor activity of bigelovin in CRC was through induction of apoptosis by up-regulating DR5 and increasing ROS. In conclusion, these results strongly suggested that bigelovin has potential to be developed as therapeutic agent for CRC patients.


Assuntos
Antineoplásicos Fitogênicos/administração & dosagem , Neoplasias Colorretais/tratamento farmacológico , Lactonas/administração & dosagem , Receptores do Ligante Indutor de Apoptose Relacionado a TNF/genética , Sesquiterpenos/administração & dosagem , Animais , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Neoplasias Colorretais/genética , Neoplasias Colorretais/patologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Células HCT116 , Células HT29 , Humanos , Camundongos , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio , Ensaios Antitumorais Modelo de Xenoenxerto
6.
Mol Med Rep ; 13(5): 3993-4000, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-27035121

RESUMO

Oxidative stress has been demonstrated to be important during myocardial ischemia/reperfusion injury (MIRI). The lazaroid U83836E, which combines the amino functionalities of the 21­aminosteroids with the antioxidant ring portion of vitamin E, is a reactive oxygen species scavenger. The aim of the current study was to investigate the effect of U83836E on MIRI and its mechanisms of action. Rat hearts were subjected to 30 min ligation of the left anterior descending coronary artery, followed by 2 h reperfusion. The results demonstrated that at 5 mg/kg, U83836E markedly protected cardiac function in ischemia/reperfusion rat models, decreased the malondialdehyde content and creatinine kinase activity, while increasing superoxide dismutase and glutathione peroxidase activity. Additionally, U83836E significantly decreased the histological damage to the myocardium, reduced the area of myocardial infarction in the left ventricle and modified the mitochondrial dysfunction. Furthermore, U83836E enhanced the translocation of protein kinase Cε (PKCε) from the cytoplasm to the membrane. However, the cardioprotective effects of U83836E were reduced in the presence of the PKC inhibitor, chelerythrine (1 mg/kg). Therefore, the results of the present study suggest that U83836E has a potent protective effect against MIRI in rat models through the direct anti­oxidative stress mechanisms and the activation of PKC signaling.


Assuntos
Cromanos/farmacologia , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Miocárdio/enzimologia , Estresse Oxidativo/efeitos dos fármacos , Piperazinas/farmacologia , Proteína Quinase C-épsilon/metabolismo , Transdução de Sinais/efeitos dos fármacos , Animais , Membrana Celular/enzimologia , Citoplasma/enzimologia , Ativação Enzimática/efeitos dos fármacos , Masculino , Traumatismo por Reperfusão Miocárdica/enzimologia , Traumatismo por Reperfusão Miocárdica/patologia , Miocárdio/patologia , Transporte Proteico/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley
7.
Int J Nanomedicine ; 10: 4239-53, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26170661

RESUMO

Malaria is still a serious public health problem in some parts of the world. The problems of recurrence and drug resistance are increasingly more serious. Thus, it is necessary to develop a novel antimalarial agent. The objectives of this study were to construct a novel compound antimalarial transdermal nanosystem-ethosomal cataplasm, to investigate its characteristics and efficiency, and to systematically explore the penetration-enhancing mechanisms of ethosomal cataplasm. Artesunate-loaded ethosomes and febrifugine-loaded ethosomes were prepared, and their characteristics were evaluated. Drug-loaded ethosomes were incorporated in the matrix of cataplasm to form the compound antimalarial ethosomal cataplasm. With the help of ethosomal technology, the accumulated permeation quantity of artesunate significantly increased at 8 hours after administration, which was 1.57 times as much as that of conventional cataplasm. Soon after administration, the ethosomal cataplasm could make a large quantity of antimalarial drug quickly penetrate through skin, then the remaining drug in the ethosomal cataplasm could be steadily released. These characteristics of ethosomal cataplasm are favorable for antimalarial drugs to kill Plasmodium spp. quickly and prevent the resurgence of Plasmodium spp. As expected, the ethosomal cataplasm showed good antimalarial efficiency in this experiment. The negative conversion rates were 100% and the recurrence rates were 0% at all dosages. The mechanism of penetration enhancement of the ethosomal cataplasm was systematically explored using an optics microscope, polarization microscope, and transmission electron microscopy. The microstructure, ultrastructure, and birefringent structure in skin were observed. Data obtained in this study showed that the application of ethosomal technology to antimalarial cataplasm could improve the transdermal delivery of drug, enhance the efficacy, and facilitate practical application in clinic.


Assuntos
Antimaláricos , Portadores de Fármacos , Nanopartículas , Administração Cutânea , Animais , Antimaláricos/administração & dosagem , Antimaláricos/química , Antimaláricos/farmacocinética , Artemisininas/administração & dosagem , Artemisininas/química , Artemisininas/farmacocinética , Artesunato , Portadores de Fármacos/administração & dosagem , Portadores de Fármacos/química , Portadores de Fármacos/farmacocinética , Etanol , Masculino , Camundongos , Nanopartículas/administração & dosagem , Nanopartículas/química , Fosfolipídeos , Pele/química , Pele/metabolismo , Tensoativos
8.
J Agric Food Chem ; 62(50): 12128-33, 2014 Dec 17.
Artigo em Inglês | MEDLINE | ID: mdl-25372841

RESUMO

Quality control issues overshadow potential health benefits of the edible mushroom Flammulina velutipes, with the detection and isolation of polysaccharides posing particular problems. In this study, multiple-fingerprint analysis was performed using chemometrics to assess polysaccharide quality and antioxidant activity of F. velutipes fruiting bodies from different sources. The authentic source exhibited differences in both oxygen radical absorbance capacity and ferric reducing antioxidant power from foreign sources. IR spectroscopic/HPLC fingerprints of polysaccharide extracts from the authentic source were established and applied to assess the polysaccharide quality of foreign sources. Analysis of IR fingerprints using Pearson correlation coefficient gave correlation coefficient r values of 0.788 and 0.828 for two foreign sources, respectively, indicating distinctness from the authentic source. Analysis of HPLC fingerprints using the supervised method by Traditional Chinese Medicine could not discriminate between sources (r > 0.9), but principal component analysis of IR and HPLC fingerprints distinguished the foreign sources.


Assuntos
Flammulina/química , Polissacarídeos/química , Cromatografia Líquida de Alta Pressão , Carpóforos/química , Controle de Qualidade
9.
Molecules ; 19(7): 9675-88, 2014 Jul 07.
Artigo em Inglês | MEDLINE | ID: mdl-25004074

RESUMO

Red radish (Raphanus L.) pickles are popular appetizers or spices in Asian-style cuisine. However, tons of radish brines are generated as wastes from industrial radish pickle production. In this study, we evaluated the dynamic changes in colour properties, phenolics, anthocyanin profiles, phenolic acid composition, flavonoids, and antioxidant properties in radish brines during lactic acid fermentation. The results showed that five flavonoids detected were four anthocyanins and one kaempferol derivative, including pelargonidin-3-digluoside-5-glucoside derivatives acylated with p-coumaric acid, ferulic acid, p-coumaric and manolic acids, or ferulic and malonic acids. Amounts ranged from 15.5-19.3 µg/mL in total monomeric anthocyanins, and kaempferol-3,7-diglycoside (15-30 µg/mL). 4-Hydroxy-benzoic, gentisic, vanillic, syringic, p-coumaric, ferulic, sinapic and salicylic acids were detected in amounts that varied from 70.2-92.2 µg/mL, whereas the total phenolic content was 206-220 µg/mL. The change in colour of the brine was associated with the accumulation of lactic acid and anthocyanins. The ORAC and Fe2+ chelation capacity of radish brines generally decreased, whereas the reducing power measured as FRAP values was increased during the fermentation from day 5 to day 14. This study provided information on the phytochemicals and the antioxidative activities of red radish fermentation waste that might lead to further utilization as nutraceuticals or natural colorants.


Assuntos
Antioxidantes/química , Fermentação , Ácido Láctico/química , Compostos Fitoquímicos/química , Raphanus/química , Sais/química , Antocianinas , Antioxidantes/farmacologia , Flavonoides , Concentração de Íons de Hidrogênio , Hidroxibenzoatos , Fenóis , Compostos Fitoquímicos/farmacologia , Pigmentos Biológicos/química , Sais/farmacologia
10.
Int Immunopharmacol ; 18(1): 1-6, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24161745

RESUMO

The aim of this study was to investigate the expression of G proteins in fibroblast-like synoviocytes (FLSs) from rats with collagen-induced arthritis (CIA) and to determine the effect of total glucosides of paeony (TGP). CIA rats were induced with chicken type II collagen (CCII) in Freund's complete adjuvant. The rats with experimental arthritis were randomly separated into five groups and then treated with TGP (25, 50, and 100mg/kg) from days 14 to 35 after immunization. The secondary inflammatory reactions were evaluated through the polyarthritis index and histopathological changes. The level of cyclic adenosine monophosphate (cAMP) was measured by radioimmunoassay. The FLS proliferation response was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The toxin-catalyzed ADP-ribosylation of G proteins was performed through autoradiography. The results show that TGP (25, 50, and 100mg/kg) significantly decreased the arthritis scores of CIA rats and improved the histopathological changes. TGP inhibited the proliferation of FLSs and increased the level of cAMP. Moreover, the FLS proliferation and the level of Gαi expression were significantly increased, but the level of Gαs expression was decreased after stimulation with IL-1ß (10ng/ml) in vitro. TGP (12.5 and 62.5µg/ml) significantly inhibited the FLS proliferation and regulated the balance between Gαi and Gαs. These results demonstrate that TGP may exert its anti-inflammatory effects through the suppression of FLS proliferation, which may be associated with its ability to regulate the balance of G proteins. Thus, TGP may have potential as a therapeutic agent for the treatment of rheumatoid arthritis.


Assuntos
Anti-Inflamatórios/administração & dosagem , Artrite Experimental/tratamento farmacológico , Artrite Reumatoide/tratamento farmacológico , Fibroblastos/efeitos dos fármacos , Proteínas de Ligação ao GTP/metabolismo , Glucosídeos/uso terapêutico , Paeonia , Fitoterapia , Extratos Vegetais/administração & dosagem , Membrana Sinovial/patologia , Animais , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Galinhas , Colágeno Tipo II/imunologia , AMP Cíclico/metabolismo , Modelos Animais de Doenças , Fibroblastos/patologia , Humanos , Masculino , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos
11.
Zhongguo Zhong Yao Za Zhi ; 39(24): 4778-81, 2014 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-25898577

RESUMO

To evaluate in vitro release and transdermal behaviors of Huoxue Zhitong gel, modified Franz diffusion cell methods was applied to investigate in vitro transdermal absorption of Huoxue Zhitong gel and the content of paeonolan in receptor fluid composed of PEG400%-95% ethanol-water (l:3:6)were determined by HPLC. The results were processed and different equations were fitted. The release law were in accordance with Weibull equation and the fitting equation was In[-1/(1 - Q)] = -0.790 51nt - 1.7012 (r = 0.9809). In 8 hours, cumulative release of paeonol was 85. 18% and the release rate was 2.827 µg . cm-2 h-1. Transdermal actions were consistent with zero-level model fit and the fitting equation was Q(t) = 1.7579t + 0. 7213 (r = 0.9991). In 8 hours, cumulative transdermal rate and transmission rate of paeonol was 54. 85%, 1. 820 µg . cm-2 h-1. So the Huoxue Zhitong gel had a good release and transdermal properties.


Assuntos
Acetofenonas/farmacocinética , Medicamentos de Ervas Chinesas/farmacocinética , Absorção Cutânea , Acetofenonas/administração & dosagem , Administração Cutânea , Animais , Cromatografia Líquida de Alta Pressão , Medicamentos de Ervas Chinesas/administração & dosagem , Géis , Camundongos
12.
Zhongguo Zhong Yao Za Zhi ; 38(14): 2306-8, 2013 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-24199560

RESUMO

To evaluate in vitro release and transdermal behaviors of Zhitong cataplasm, modified Franz diffusion cell method was applied to investigate in vitro transdermal absorption of Zhitong cataplasm and the content of tetrahydropalmatine was determined by HPLC. In 24 hours, accumulative release rate of tetrahydropalmatine was 81. 9%, transmission rate was 2.26 microg x cm(-2) x h(-1). In 48 hours, accumulative transdermal rate and transmission rate of tetrahydropalmatine were 20.31%, 0.22 pg x cm(-2) x h(-1). So Zhitong cataplasm had a good release and transdermal properties and transdermal actions were consistent with zero-order kinetics process.


Assuntos
Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/farmacocinética , Administração Cutânea , Animais , Alcaloides de Berberina/administração & dosagem , Alcaloides de Berberina/química , Alcaloides de Berberina/farmacocinética , Medicamentos de Ervas Chinesas/administração & dosagem , Masculino , Camundongos , Extratos Vegetais/administração & dosagem , Extratos Vegetais/química , Extratos Vegetais/farmacocinética , Pele/metabolismo , Absorção Cutânea
13.
Zhonghua Zhong Liu Za Zhi ; 35(5): 347-50, 2013 May.
Artigo em Chinês | MEDLINE | ID: mdl-24054010

RESUMO

OBJECTIVE: To establish a quantitative method to detect circulating tumor cells (CTC) in patients with small cell lung cancer, and analyze its sensitivity and stability. METHODS: A specific primer and probe for prepro-gastrin-releasing peptide (preproGRP) was designed and a quantitative RT-PCR method was established to detect preproGRP mRNA. Cell incorporation method was used to evaluate the sensitivity. Magnetic cell sorting (MACS) was used to isolate and purify CTC from peripheral blood, and the MACS in combination with morphological diagnosis were used for cell counting. RESULTS: The isolation rate of CTC by MACS was 30% and the lower detection limit was 5 cells per ml blood. The sensitivity of quantitative RT-PCR in detection of preproGRP mRNA in CTC was 0.64 cells per reaction, and the lower detection limit was 50 cells per ml blood, which was lower than that of MACS. However, the cell numbers calculated by Ct value was in greater accordance (about 80%) with actual cell numbers than that obtained by MACS. CONCLUSIONS: PreproGRP quantitative RT-PCR and MACS have both advantages and disadvantages in detecting CTC of SCLC patients. MACS has a higher sensitivity, and is more favorable when CTC count is below 50 per ml blood. Meanwhile, preproGRP mRNA quantitative RT-PCR is more reliable in calculating actual cell numbers.


Assuntos
Neoplasias Pulmonares/patologia , Células Neoplásicas Circulantes , Peptídeos/metabolismo , Precursores de Proteínas/metabolismo , Carcinoma de Pequenas Células do Pulmão/patologia , Humanos , Separação Imunomagnética , Neoplasias Pulmonares/sangue , Neoplasias Pulmonares/metabolismo , Peptídeos/genética , Precursores de Proteínas/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Carcinoma de Pequenas Células do Pulmão/sangue , Carcinoma de Pequenas Células do Pulmão/metabolismo
14.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 28(11): 1177-81, 2012 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-23127409

RESUMO

AIM: To prepare and characterize monoclonal antibodies against matrix metalloproteinase-2 (MMP-2), check its expression in the tissues of human ovarian cancer and transplanted tumors in nude mice. METHODS: MMP-2 were linked to the carrier protein bovine serumalbumin (BSA) and keyhole limpet hemocyanin (KLH) using glutaraldehyde method to obtain MMP-2-BSA and MMP-2-KLH, respectively. The anti-MMP-2 monoclonal antibody was obtained through hybridoma technique. We established the cell strains secreting mAb by hybridoma technique and prepared the mAb by induction of ascites in vivo. The prepared mAb was purified by salting out with ammonium sulfate and identified by ELISA and Western blotting. We compared the mAb and commercial polyclonal antibody by immunohistochemistry and detected the expressions of MMP-2 and CA125 in ovarian cancer issues and transplanted tumor. RESULTS: The artificial antigen and 3 hybridoma cell lines secreting monoclonal antibodies (mAb) against MMP-2 were obtained. The subclasses of mAb were all IgG1. The titer of peritoneal exudates was 1:1×10(6);. The expressions of MMP-2 and CA125 in transplanted tumor and ovarian cancer tissues were all high. The positive expression rate of MMP-2 checked using generated antibody was 71.2%(57/80) in ovarian cancer tissues and 16.67% (5/30) in normal tissues, with significant difference between them (P<0.01). In early stage, the positive rate of MMP-2 and CA125 combined detection was higher than that of CA125 detection alone (P<0.01). The mAb was suitable for detecting the expression of MMP-2 in human tissues and gave results consistent with commercial polyclonal antibody. The mAb was more specific than commercial mAb (P<0.01). CONCLUSION: The anti-human MMP-2 mAb is successfully prepared, which may serve as a valuable tool in the functionaI studies of ovarian cancer.


Assuntos
Anticorpos Monoclonais/biossíntese , Metaloproteinase 2 da Matriz/imunologia , Animais , Anticorpos Monoclonais/imunologia , Antígeno Ca-125/análise , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Metaloproteinase 2 da Matriz/análise , Camundongos , Camundongos Endogâmicos BALB C , Transplante de Neoplasias , Neoplasias Ovarianas/química , Neoplasias Ovarianas/enzimologia , Ratos , Transplante Heterólogo
15.
J Pineal Res ; 53(4): 325-34, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22537289

RESUMO

Increasing evidence demonstrates that melatonin has an anti-inflammatory effect. Nevertheless, the molecular mechanisms remain obscure. In this study, we investigated the effect of melatonin on toll-like receptor 4 (TLR4)-mediated molecule myeloid differentiation factor 88 (MyD88)-dependent and TRIF-dependent signaling pathways in lipopolysaccharide (LPS)-stimulated macrophages. RAW264.7 cells were incubated with LPS (2.0 µg/mL) in the absence or presence of melatonin (10, 100, 1000 µm). As expected, melatonin inhibited TLR4-mediated tumor necrosis factor alpha (TNF-α), interleukin (IL)-1ß, IL-6, IL-8, and IL-10 in LPS-stimulated macrophages. In addition, melatonin significantly attenuated LPS-induced upregulation of cyclooxygenase (COX)-2 and inducible nitric oxide synthase (iNOS) in macrophages. Further analysis showed that melatonin inhibited the expression of MyD88 in LPS-stimulated macrophages. Although it had no effect on TLR4-mediated phosphorylation of c-Jun N-terminal kinase (JNK), p38, and extracellular regulated protein kinase (ERK), melatonin significantly attenuated the activation of nuclear factor kappa B (NF-κB) in LPS-stimulated macrophages. In addition, melatonin inhibited TLR4-mediated Akt phosphorylation in LPS-stimulated macrophages. Moreover, melatonin significantly attenuated the elevation of interferon (IFN)-regulated factor-3 (IRF3), which was involved in TLR4-mediated TRIF-dependent signaling pathway, in LPS-stimulated macrophages. Correspondingly, melatonin significantly alleviated LPS-induced IFN-ß in macrophages. In conclusion, melatonin modulates TLR4-mediated inflammatory genes through MyD88-dependent and TRIF-dependent signaling pathways.


Assuntos
Proteínas Adaptadoras de Transporte Vesicular/metabolismo , Anti-Inflamatórios/farmacologia , Mediadores da Inflamação/metabolismo , Inflamação/prevenção & controle , Lipopolissacarídeos/farmacologia , Macrófagos/efeitos dos fármacos , Melatonina/farmacologia , Fator 88 de Diferenciação Mieloide/metabolismo , Transdução de Sinais/efeitos dos fármacos , Receptor 4 Toll-Like/efeitos dos fármacos , Proteínas Adaptadoras de Transporte Vesicular/genética , Animais , Linhagem Celular , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/metabolismo , Citocinas/genética , Citocinas/metabolismo , Relação Dose-Resposta a Droga , Ativação Enzimática , Regulação da Expressão Gênica , Inflamação/genética , Inflamação/imunologia , Inflamação/metabolismo , Macrófagos/imunologia , Macrófagos/metabolismo , Camundongos , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Fator 88 de Diferenciação Mieloide/genética , NF-kappa B/metabolismo , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo II/metabolismo , Fosforilação , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Mensageiro/metabolismo , Fatores de Tempo , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/metabolismo
16.
Pharmazie ; 67(12): 997-1001, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23346762

RESUMO

An investigation was designed and conducted to detect pharmacokinetic differences between paeoniflorin (Pae) microemulsion and Pae saline. Pae microemulsion (25, 50,100 mg x kg(-1)) was administered to three groups of rats with adjuvant arthritis (AA) while Pae (25, 50,100 mg x kg(-1)) was given to another three groups of rats both for ten days. A HPLC assay was developed to determine the plasma concentrations of Pae. The plasma concentrations of Pae groups (25, 50 mg x kg(-1)) were undetectable. Furthermore, compared with pharmacokinetic parameters of Pae group (100 mg x kg(-1)), maximum concentration (C(max)), the area under the plasma concentration-time curve (AUC(0-t)), and mean retention time MRT(0-infinity))(h) of Pae microemulsion (100 mg x kg(-1)) increased apparently, while volume of distribution (Vd) and clearance rate (CL/F) decreased. These results indicate that a microemulsion significantly improves the absorption of Pae in AA rats.


Assuntos
Anti-Inflamatórios não Esteroides/farmacocinética , Artrite Experimental/metabolismo , Benzoatos/farmacocinética , Hidrocarbonetos Aromáticos com Pontes/farmacocinética , Glucosídeos/farmacocinética , Animais , Anti-Inflamatórios não Esteroides/administração & dosagem , Anti-Inflamatórios não Esteroides/uso terapêutico , Área Sob a Curva , Artrite Experimental/tratamento farmacológico , Artrite Experimental/patologia , Benzoatos/administração & dosagem , Benzoatos/uso terapêutico , Disponibilidade Biológica , Hidrocarbonetos Aromáticos com Pontes/administração & dosagem , Hidrocarbonetos Aromáticos com Pontes/uso terapêutico , Calibragem , Cromatografia Líquida de Alta Pressão , Edema/etiologia , Edema/patologia , Emulsões , Pé/patologia , Adjuvante de Freund , Glucosídeos/administração & dosagem , Glucosídeos/uso terapêutico , Meia-Vida , Indicadores e Reagentes , Masculino , Monoterpenos , Paeonia/química , Raízes de Plantas/química , Ratos , Ratos Sprague-Dawley
17.
Zhonghua Zhong Liu Za Zhi ; 32(11): 850-4, 2010 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-21223692

RESUMO

OBJECTIVE: To investigate the value of carcinoembryonic antigen (CEA) or cytokeratin 19 fragment (CYFRA21-1) as an assessment indicator of therapeutic efficacy in advanced non-small cell lung cancer (NSCLC) patients. METHODS: 228 cases of advanced NSCLC with chemotherapy were enrolled into this retrospective study. The serum CEA or CYFRA21-1 levels of all patients were above the cut-off limit before treatment. The relationship between changes of tumor markers (TMs) and imaging therapeutic efficacy or progression-free survival (PFS) was analyzed, and the value of TMs in therapeutic efficacy assessment was evaluated. RESULTS: According to RECIST criteria, partial response (PR) occurred in 40 cases, stable disease (SD) in 151 and PD (progressive disease) in 37. The cut-off values of the changes of TMs between pre- and post-treatment were determined according to the above mentioned criteria. The CEA down (D), stable (S), above (A) groups were 90, 49 and 66 cases, respectively. CYFRA21-1 down (D), stable (S), above (A) groups were 84, 26 and 37 cases, respectively. PR groups were 68.4% and 88.9% in CEA and cyfra21-1 down groups, respectively, 7.9% and 5.6% in the above groups, respectively. PD groups were 59.4% and 76.2% in CEA and CYFRA21-1 above groups, respectively. No PD cases were in the down groups. The changes of TMs in SD group were between them. Statistically significant correlations were observed between changes of TMs and imaging therapeutic efficacy (r(CEA) = 0.45, P = 0.00; r(CYFRA21-1) = 0.44, P = 0.00). PFS among different TMs groups were significantly different (all P < 0.05), which can be used to further distinguish the prognosis among SD subgroups. CONCLUSION: Changes of TMs can be used to predict the imaging therapeutic effect and PFS of the patients, and if the SD group is divided into subgroups according to different therapeutic efficacy and prognosis, it may help the patients to receive individualized treatment.


Assuntos
Antígenos de Neoplasias/sangue , Antígeno Carcinoembrionário/sangue , Carcinoma Pulmonar de Células não Pequenas/sangue , Queratina-19/sangue , Neoplasias Pulmonares/sangue , Idoso , Biomarcadores Tumorais/sangue , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Carcinoma Pulmonar de Células não Pequenas/patologia , Progressão da Doença , Intervalo Livre de Doença , Feminino , Seguimentos , Humanos , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/patologia , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Indução de Remissão , Estudos Retrospectivos
18.
Acta Crystallogr Sect E Struct Rep Online ; 65(Pt 2): o356, 2009 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-21581955

RESUMO

The asymmetric unit of the title compound, C(7)H(8)N(4)S, contains two independent mol-ecules with slightly different conformations; the dihedral angles between the pyridine ring and mean plane of the thio-semicarbazone unit in the two mol-ecules are 2.88 (5) and 6.30 (5)°. Inter-molecular N-H⋯N and N-H⋯S hydrogen bonds link the mol-ecules into layers parallel to the ab plane.

19.
Vet Microbiol ; 127(1-2): 73-8, 2008 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-17897793

RESUMO

To assess the prevalence of antimicrobial resistance and class I integrons in Escherichia coli strains (n=58) isolated from bovine mastitis in Inner Mongolia, antimicrobial susceptibility and the presence of various types of integrons were characterized. Most isolates were susceptible to amikacin, colistin, ceftazidime, gentamicin and kanamycin, while those also exhibited high resistant incidence rates to ampicillin, amoxicillin, sulfadiazine and sulfamethoxydiazine. The integrase gene of integrons was amplified by PCR using degenerate primers. The integrons were confirmed by restriction fragment length polymorphism (RFLP) analysis of positive PCR products. Neither class II nor class III integron was detected, while 56.90% (n=33) of the isolates were positive for the presence of intI1 gene. Sequencing analysis of gene cassettes revealed that seven gene cassettes were found, which encoded resistance to trimethoprim (dfrA1 and dfrA17), aminoglycosides (aacA4, aadA1 and aadA5) and chloramphenicol (catB3), respectively. Of them, the gene cassette array dfrA17-aadA5 was found most prevalent (62.96%). The percentage of positive-integron among the isolates whose resistant profile was relatively broad (n> or =7) is 100.00%, while the one in narrow-profile isolates (n=2-6) is 30.56%. The correlation analysis revealed the incidence of integrons among the isolates were highly related to the resistant profile, indicating integrons play an important role in the dissemination and spread of the antimicrobial resistant strains.


Assuntos
Anti-Infecciosos/farmacologia , Farmacorresistência Bacteriana/genética , Escherichia coli/efeitos dos fármacos , Escherichia coli/genética , Integrons/genética , Mastite Bovina/microbiologia , Animais , Bovinos , China , Escherichia coli/isolamento & purificação , Feminino , Integrases/genética , Testes de Sensibilidade Microbiana , Dados de Sequência Molecular
20.
Artigo em Chinês | MEDLINE | ID: mdl-17971927

RESUMO

OBJECTIVE: To study a newly isolated domestic mammalian reovirus, BYD1, its ability to induce apoptosis analyze the three-dimensional structure of its major membrane penetration protein to predict its function in inducing apoptosis. METHODS: HeLa cells infected with BYD1 reovirus were metered with flow cytometer (FCM) to quantify the ratio of apoptotic cells. The data were analyzed with Student's t-test to judge the ability of BYD1 strain to induce apoptosis. The primary sequence ranged from 582 to 675 per microliter protein of BYD1, T1L, T2J and T3D were aligned and compared. The three-dimensional comparative protein structure model of microliter protein was generated by homology-modeling pipeline SWISS MODEL was applied to annotate its secondary and tertiary structure. RESULTS: BYD1 strain was verified with the ability to induce the apoptosis of HeLa cells. The 643-675 segment composing an alpha-helix showed major difference compared with prototype T2J. CONCLUSION: The newly isolated reovirus BYD1 is an apoptosis inducing strain. The alpha-helix (residues 643 to 675) of microliter protein of BYD1 may play a key role to induce the proapoptotic activity of infected cells.


Assuntos
Apoptose , Reoviridae/fisiologia , Proteínas Virais Reguladoras e Acessórias/fisiologia , Proteínas Reguladoras de Apoptose/química , Proteínas Reguladoras de Apoptose/genética , Proteínas Reguladoras de Apoptose/fisiologia , Diferenciação Celular , Feminino , Citometria de Fluxo , Células HeLa , Interações Hospedeiro-Patógeno , Humanos , Modelos Moleculares , Orthoreovirus de Mamíferos/genética , Orthoreovirus de Mamíferos/metabolismo , Orthoreovirus de Mamíferos/fisiologia , Conformação Proteica , Estrutura Terciária de Proteína , Reoviridae/genética , Reoviridae/metabolismo , Neoplasias do Colo do Útero/patologia , Neoplasias do Colo do Útero/virologia , Proteínas Virais Reguladoras e Acessórias/química , Proteínas Virais Reguladoras e Acessórias/genética
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