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1.
Microb Drug Resist ; 2020 Sep 14.
Artigo em Inglês | MEDLINE | ID: mdl-32924788

RESUMO

Objective: Antibiotic resistance of Pseudomonas aeruginosa (PA) that lowers the effectiveness of current treatments for pneumonia is a growing problem. Qi Gui Yin is a Chinese herbal medicine that has been used to improve the efficacy of antibiotic therapy against antibiotic-resistant bacteria. This study aimed to elucidate the mechanism by which Qi Gui Yin inhibits antibiotic resistance of PA. Methods: Active components of Qi Gui Yin were analyzed by chromatography. Isobaric Tags for Relative and Absolute Quantification (iTRAQ) technology was used to compare protein expression profiles of PA strains cultured in serum from rats that were and were not treated with Qi Gui Yin. Quantitative polymerase chain reaction (qPCR) analysis was performed to detect gene expression changes. Results: Proteomic analysis identified 76 differentially expressed proteins between PA strains cultured in serum from rats that were or were not treated with Qi Gui Yin. Bioinformatics analysis revealed that the largest number of differentially expressed proteins were associated with resistance mechanisms such as quorum sensing, bacterial biofilm formation, and active pumping. In addition, qPCR analysis confirmed that downregulation of iscU and arcA gene expression was associated with Qi Gui Yin treatment. Conclusions: Serum from Qi Gui Yin-treated rats could effectively inhibit antibiotic resistance of PA. Chlorogenic acid and astragaloside IV are the main components of Qi Gui Yin, which may mediate inhibition of antibiotic resistance. Our findings provide new insights into strategies involving Chinese herbal medicine that can be used to treat pneumonia caused by antibiotic-resistant bacteria.

2.
J Cell Mol Med ; 23(9): 5868-5875, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31350813

RESUMO

Gram-negative bacteria (GNB) emerge as important pathogens causing pulmonary infection, which can develop into sepsis due to bacterial resistance to antibiotics. GNB pneumonia poses a huge social and economic burden all over the world. During GNB infection in the lung, Toll-like receptor 4 (TLR4) can form a complex with MD2 and CD14 after recognizing lipopolysaccharide of GNB, initiate the MyD88- and TRIF-dependent signalling pathways and stimulate host non-specific immune response. In this review, we summarize recent progress in our understanding of the role of TLR4 in GNB pneumonia. The latest experimental results, especially in TLR4 knockout animals, suggest a promising potential of targeting TLR4 signalling pathway for the treatment of GNB pneumonia. Furthermore, we highlight the benefits of Traditional Chinese Medicine as novel candidates for the therapy of GNB pneumonia due to the modulation of TLR4 signalling pathway. Finally, we discuss the promise and challenge in the development of TLR4-based drugs for GNB pneumonia.


Assuntos
Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Medicina Tradicional Chinesa/métodos , Pneumonia Bacteriana/tratamento farmacológico , Receptor 4 Toll-Like/efeitos dos fármacos , Proteínas Adaptadoras de Transporte Vesicular/imunologia , Bactérias Gram-Negativas/metabolismo , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/microbiologia , Humanos , Lipopolissacarídeos/metabolismo , Fator 88 de Diferenciação Mieloide/imunologia , Pneumonia Bacteriana/microbiologia , Sepse/imunologia , Sepse/patologia , Transdução de Sinais/imunologia , Receptor 4 Toll-Like/agonistas , Receptor 4 Toll-Like/antagonistas & inibidores , Receptor 4 Toll-Like/metabolismo
3.
J Cell Mol Med ; 21(6): 1046-1057, 2017 06.
Artigo em Inglês | MEDLINE | ID: mdl-28039939

RESUMO

Lung diseases remain a serious problem for public health. The immune status of the body is considered to be the main influencing factor for the progression of lung diseases. HMGB1 (high-mobility group box 1) emerges as an important molecule of the body immune network. Accumulating data have demonstrated that HMGB1 is crucially implicated in lung diseases and acts as independent biomarker and therapeutic target for related lung diseases. This review provides an overview of updated understanding of HMGB1 structure, release styles, receptors and function. Furthermore, we discuss the potential role of HMGB1 in a variety of lung diseases. Further exploration of molecular mechanisms underlying the function of HMGB1 in lung diseases will provide novel preventive and therapeutic strategies for lung diseases.


Assuntos
Biomarcadores , Proteína HMGB1/genética , Pneumopatias/genética , Humanos , Pulmão/metabolismo , Pulmão/patologia , Pneumopatias/patologia , Terapia de Alvo Molecular , Receptores Imunológicos/genética
4.
Oncotarget ; 8(64): 108108-108117, 2017 Dec 08.
Artigo em Inglês | MEDLINE | ID: mdl-29296227

RESUMO

Interleukin 15 (IL-15) is a cytokine exhibiting antitumor characteristic similar to that of IL-2. However, in human tissues and cells, IL-15 expression and secretion is very limited, suggesting IL-15 functions mainly intracellularly. In the present study, we assessed the effects of transfecting NCI-H446 small cell lung cancer cells with genes encoding three IL-15 variants: prototypical IL-15, mature IL-15 peptide, and modified IL-15 in which the IL-2 signal peptide is substituted for the native signal peptide. NCI-H446 cells transfected with empty plasmid served as the control group. We found that IL-15 transfection effectively inhibited NCI-H446 cell proliferation and arrested cell cycle progression, with the modified IL-15 carrying the IL-2 signal peptide exerting the greatest effect. Consistent with those findings, expression each of the three IL-15 variants reduced growth of NCI-H446 xenograph tumors, and the modified IL-15 again showed the greatest effect. In addition, IL-15 expression led to down-regulation of the positive cell cycle regulators cyclin E and CDK2 and up-regulation of the negative cycle regulators p21 and Rb. These findings suggest IL-15 acts as a tumor suppressor that inhibits tumor cell proliferation by inducing cell cycle arrest.

5.
Mediators Inflamm ; 2016: 8172706, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27433030

RESUMO

Hydroxysafflor yellow A (HSYA) is an effective therapeutic agent for inflammatory diseases and autoimmune disorders; however, its regulatory effect on NLRP3 inflammasome activation in macrophages has not been investigated. In this study, we predicted the potential interaction between HSYA and xanthine oxidase (XO) via PharmMapper inverse docking and confirmed the binding inhibition via inhibitory test (IC50 = 40.04 µM). Computation docking illustrated that, in this HSYA-XO complex, HSYA was surrounded by Leu 648, Leu 712, His 875, Leu 873, Ser 876, Glu 879, Phe 649, and Asn 650 with a binding energy of -5.77 kcal/M and formed hydrogen bonds with the hydroxyl groups of HSYA at Glu 879, Asn 650, and His 875. We then found that HSYA significantly decreased the activity of XO in RAW264.7 macrophages and suppressed LPS-induced ROS generation. Moreover, we proved that HSYA markedly inhibited LPS-induced cleaved caspase-1 activation via suppressing the sensitization of NLRP3 inflammasome and prevented the mature IL-1ß formation from pro-IL-1ß form. These findings suggest that XO may be a potential target of HSYA via direct binding inhibition and the combination of HSYA-XO suppresses LPS-induced ROS generation, contributing to the depression of NLRP3 inflammasome and inhibition of IL-1ß secretion in macrophages.


Assuntos
Chalcona/análogos & derivados , Inflamassomos/metabolismo , Macrófagos/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Quinonas/química , Xantina Oxidase/metabolismo , Animais , Chalcona/química , Simulação por Computador , Concentração Inibidora 50 , Interleucina-1beta/metabolismo , Lipopolissacarídeos , Camundongos , Pigmentos Biológicos/química , Células RAW 264.7 , Espécies Reativas de Oxigênio/metabolismo
6.
PLoS One ; 10(2): e0117736, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25689855

RESUMO

BACKGROUND: Hepatitis B vaccine that contains an aluminum hydroxide adjuvant induces apoptotic death of Hepa 1-6 cells. Difficult-to-degrade chemical additives in vaccines effectively enhance vaccine immunogenicity, but also affect the host tissue. Identification of bio-molecules that are readily degraded and compatible in vivo as an adjuvant is important for vaccine research. The hapten-carrier effect suggests that stimulation of helper T (Th) cells by carrier adjuvants is feasible. Protein D (PD) of non-typeable Haemophilus influenzae covalently conjugated to some polysaccharide vaccines has been confirmed to convert T-cell independent (TI) antigens into T-cell dependent (TD) antigens, and elicit strong T-cell responses ultimately. Herein, we would substitube PD for aluminum hydroxide adjuvant in Hepatitis B vaccine. METHODS AND RESULTS: Truncated PD (amino acids 20-364) was expressed in Escherichia coli and purified by (NH4)2SO4 precipitation and DEAE chromatography. After evaluation of antigenicity by western blotting, PD was covalently conjugated to yeast-derived recombinant HBsAg by cross-linking with glutaraldehyde. Intramuscular immunization with the conjugate induced higher level of HBsAg-specific antibody than did HBsAg alone (p < 0.05), and was comparable to commercial Hepatitis B vaccine. During the surveillance period (days 35-105), anti-HBs titers were hold high. Moreover, the conjugated vaccine enhanced Th1 immune responses, while Th2 responses were also activated and induced an antibody response, as determined by IFN-γ ELISPOT and IgG1/IgG2a ratio assays. CONCLUSIONS: Recombinant truncated PD covalently conjugated to HBsAg antigen enhanced the immunogenicity of the antigen in mice simultaneously by humoral and cellular immune response, which would facilitate therapeutic hepatitis B vaccines.


Assuntos
Proteínas de Bactérias/metabolismo , Proteínas de Transporte/metabolismo , Haemophilus influenzae/imunologia , Antígenos de Superfície da Hepatite B/imunologia , Imunoconjugados/imunologia , Imunoglobulina D/metabolismo , Lipoproteínas/metabolismo , Animais , Anticorpos Antivirais/imunologia , Linfócitos B/citologia , Linfócitos B/imunologia , Proteínas de Bactérias/genética , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Proteínas de Transporte/genética , Comunicação Celular/imunologia , Feminino , Haemophilus influenzae/classificação , Antígenos de Superfície da Hepatite B/metabolismo , Imunoglobulina D/genética , Imunoglobulina G/imunologia , Lipoproteínas/genética , Camundongos , Plasmídeos/genética , Baço/imunologia , Linfócitos T Auxiliares-Indutores/citologia , Linfócitos T Auxiliares-Indutores/imunologia , Vacinas Virais/imunologia
7.
J Virol Methods ; 197: 34-8, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24140186

RESUMO

Hepatitis D virus (HDV) infection is often accompanied by hepatitis B virus (HBV) infection. Co-infection of HDV and HBV may lead to more severe symptoms and even death. Current methods for HDV diagnosis have high false-positive rates and show significant result discrepancies. The Abbott AxSYM AUSAB test, currently a standard test for HDV detection, is too laborious and expensive for routine application. Therefore, new sensitive and cost-efficient methods for HDV diagnosis are urgently needed. In this study, S-HDAg protein was produced in high yield in Escherichia coli. Optimal protein production was achieved by optimization of S-HDAg gene codons according to the codon preference of E. coli and using host cells with appropriate cell density. Under optimal expression conditions, the S-HDAg protein expression yield (30mg/l) was the highest among any proteins expressed in E. coli. A standard enzyme-linked immunosorbent assay (ELISA) for HDV was developed using the purified S-HDAg protein, which showed high specificity against hepatitis B, C, D and E viruses. Overall, the ELISA had superior specificity and sensitivity compared with the Abbott AxSYM AUSAB test and was also more convenient and cost-efficient.


Assuntos
Anticorpos Anti-Hepatite/sangue , Hepatite D/diagnóstico , Vírus Delta da Hepatite/imunologia , Antígenos da Hepatite delta , Clonagem Molecular , Ensaio de Imunoadsorção Enzimática/métodos , Escherichia coli/genética , Expressão Gênica , Antígenos da Hepatite delta/genética , Humanos , Imunoglobulina G/sangue , Proteínas Recombinantes/genética , Sensibilidade e Especificidade
8.
Artigo em Chinês | MEDLINE | ID: mdl-23002540

RESUMO

OBJECTIVE: To prepare HDAg with biological activities as a candidate of diagnostic reagent. METHODS: To synthesize HDV gene fragment after codon optimization. To construct a thio-fused recombinant plasmid based on M48 expression vector. To express in E. coli induced by IPTG. To purify the protein by affinity chromatography followed by characterization in ELISA: RESULTS: Plasmid construction was verified by enzyme digestion. SDS-PAGE indicated the molecular weight of the protein was the same as we expectation. ELISA proved its affinity with HDV antibodies. CONCLUSION: HDAg was obtained successfully and it will pave the road to the research of HDV diagnostic reagent.


Assuntos
Hepatite D/diagnóstico , Antígenos da Hepatite delta/imunologia , Ensaio de Imunoadsorção Enzimática , Antígenos da Hepatite delta/genética , Antígenos da Hepatite delta/isolamento & purificação , Humanos , Plasmídeos , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/isolamento & purificação
9.
J Hazard Mater ; 235-236: 85-91, 2012 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-22841801

RESUMO

In situ transformation of 4,4'-Dibromobiphenyl (4,4'-DBB) in water was observed with hydrothermal diamond anvil cell (HDAC) up to 633 K. It shows that 4,4'-DBB dissolves in water to form a homogenous phase at the temperature of 588 K and thus subcritical water oxidation of 4,4'-DBB higher than the temperature can be a homogenous phase. To accelerate the oxidative degradation, some Mn-Ce-Co complex oxide nanoparticles of about 100 nm were prepared by co-precipitation hydrothermal method. The nanoparticles show enough stability and catalytic activity for oxidative degradation of 4,4'-DBB in subcritical water. The catalytic activation increases with some Co doping and as for the complex oxides of Mn(1)Ce(1), Mn(0.9)Ce(1)Co(0.1), Mn(0.5)Ce(1)Co(0.5), Mn(0.1)Ce(1)Co(0.9), and Co(1)Ce(1), the Mn(0.9)Ce(1)Co(0.1) presents the best activation. The main intermediate products of degradation are benzoic acid and phenol. The apparent activation energy (E(a)) is 35.92 with 5% Mn(0.9)Ce(1)Co(0.1) as catalyst and 46.69 kJ/mol with no catalyst about the chemical oxygen demand (COD).


Assuntos
Compostos de Bifenilo/química , Nanopartículas Metálicas/química , Poluentes Químicos da Água/química , Catálise , Cério/química , Cobalto/química , Temperatura Alta , Manganês/química , Oxirredução , Óxidos/química , Purificação da Água/métodos
10.
Artigo em Chinês | MEDLINE | ID: mdl-23547452

RESUMO

OBJECTIVE: To study the effect of gene optimization on the expression and purification of HDV small antigen produced by genetic engineering. METHODS: Based on the colon preference of E. coli, the HDV small antigen original gene from GenBank was optimized. Both the original gene and the optimized gene expressed in prokaryotic cells, SDS-PAGE was made to analyze the protein expression yield and to decide which protein expression style was more proportion than the other. Furthermore, two antigens were purified by chromatography in order to compare the purity by SDS-PAGE and Image Lab software. RESULTS: SDS-PAGE indicated that the molecular weight of target proteins from two groups were the same as we expected. Gene optimization resulted in the higher yield and it could make the product more soluble. After chromatography, the purity of target protein from optimized gene was up to 96.3%, obviously purer than that from original gene. CONCLUSION: Gene optimization could increase the protein expression yield and solubility of genetic engineering HDV small antigen. In addition, the product from the optimized gene group was easier to be purified for diagnosis usage.


Assuntos
Engenharia Genética/métodos , Antígenos da Hepatite delta/genética , Proteínas Recombinantes/biossíntese , Eletroforese em Gel de Poliacrilamida , Hepatite D/diagnóstico , Antígenos da Hepatite delta/isolamento & purificação , Proteínas Recombinantes/isolamento & purificação
11.
Artigo em Chinês | MEDLINE | ID: mdl-23547464

RESUMO

OBJECTIVE: To investigate the seroprevalence of hepatitis D virus in Foshan of Guangdong province, to provide the data for the study about it in China. METHODS: ELISA kits from two different companies were used for detecting anti-HDV IgG of all the serum samples, and then RT-PCR was carried out about the selected serum to ensure the results. All the serum samples were collected in 2011 in The First People's Hospital of Foshan. RESULTS: The results from two ELISA kits and RT-PCR were identical. Eight samples were positive. CONCLUSIONS: The seroprevalence rate of HDV in Foshan is higher than that in China. It has no statistically significant difference between female and male. Morever, the older with HBsAg are susceptible to HDV.


Assuntos
Coinfecção/epidemiologia , Antígenos de Superfície da Hepatite B/sangue , Hepatite B/epidemiologia , Hepatite D/epidemiologia , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , China/epidemiologia , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase Via Transcriptase Reversa
12.
Artigo em Chinês | MEDLINE | ID: mdl-23627037

RESUMO

OBJECTIVE: To establish a novel improved loop-mediated isothermal amplification (LAMP) technique to detect hepatitis A virus (HAV). METHODS: A novel improved LAMP assay based on the addition of an acceleration primer was developed for hepatitis A virus nucleotide acid detection. RESULTS: Precision and reproducibility analysis proved its high stability and reliability. Comparison between the improved and conventional LAMP assays revealed that the former was more sensitive with a detection limit of 5 TCID50/ml. The novel detection method displayed 100% consistency with the TaqMan real-time PCR assay when applied to clinical specimens collected from patients with confirmed acute HAV infection. CONCLUSION: This novel technique is widely applicable as a simple diagnostic tool in the clinical field as well as for the surveillance and investigation of the infectious disease in developing countries where HAV is endemic.


Assuntos
Vírus da Hepatite A/isolamento & purificação , Hepatite A/virologia , Técnicas de Amplificação de Ácido Nucleico/métodos , RNA Viral/genética , Primers do DNA/genética , Vírus da Hepatite A/genética , Humanos , RNA Viral/metabolismo
13.
Guang Pu Xue Yu Guang Pu Fen Xi ; 29(8): 2112-6, 2009 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-19839320

RESUMO

FeS2 belongs to sulfide, including pyrite of isometric system and marcasite of orthorhombic system. The FeS2 discovered in Gengzhuang, Shanxi Province, was growing in the form of whisker. The study with scanning electron microscopy and electron probe show that the mineral components of FeS2 vary regularly. The structure of natural nano-micron FeS2 whisker was determined by micro-Raman spectroscopy. The results show that there exist two types of structure in FeS2 whiskers: pyrite and marcasite. Marcasite presents irregular shapes, such as coarse lotus root joints, crude columnar or beaded. Pyrite exists in the shape of straight line and smooth surface. In the early growing stage, Gengzhuang FeS2 whisker was mainly marcasite-type structure; in the middle stage it was coexistent structure of pyrite- and marcasite-type; in the late stage it was mainly pyrite-type. The growing stages of the whisker FeS2 show the phase transformation laws. Moreover, during the growing process marcasite was growing with pyrite coated on. Study on FeS2 whisker structure shows that there are correlations between phase transformation laws of the structure and forms, and between the forming time and the composition characteristics.

14.
Zhongguo Zhong Yao Za Zhi ; 34(2): 212-6, 2009 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-19385189

RESUMO

OBJECTIVE: To investigate the dynamic changes in angiogenesis within the tumor tissue of mice bearing S180 tumor at different day-points of oral administration with a Chinese medicine compound "Yiliuyin" (YLY) and to explore the anti-tumor mechanisms of YLY in vivo. METHOD: Fifty-six BALB/c mice were divided into YLY group and control group (28 mice/group) and each group was divided into four subgroups (7 mice/subgroup), randomly. After 24 hrs of inoculation with S180 tumor cells subcutaneously in the right axilla, YLY in the mice of YLY group and equal volume of cold boiled-water in the mice of control group were administered orally twice every day, 0.5 mL each time. The mice of one subgroup from the two groups apiece were killed at 10, 20, 30 th and 40 th day-point of oral administration, respectively. The tumors were isolated and were made into paraffin embedded sections. The dynamic changes of the angiogenesis (CD34 staining), vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor-2 (VEGFR-2) and endostatin (ES) in tumor tissue were detected by immunohistochemistry staining, and the results were shown as PED (positive enzyme dot). RESULT: YLY could remarkably decrease the angiogenesis within tumor tissues. The PED of CD34 in control group at 10, 20, 30 th and 40 th day-point was 392.86+/-42.01, 481.49+/-58.34, 386.31+/-54.91 and 376.69+/-28.71, and that in YLY group was 334.46+/-33.38, 289.34+/-39.63, 257.09+/-40.00 and 246.57+/-36.78, respectively. The PED of CD34 in YLY group at each day-point was lower than that in control group (P<0.05, P<0.01, P<0.01 and P<0.01, respectively). The PED of VEGF in control group at 10, 20, 30 th and 40 th day-point was 852.63+/-81.65, 1168.40+/-96.69, 1292.60+/-147.54 and 1124.74+/-139.64, and that inYLY group was 718.40+/-94.94, 866.54+/-72.40, 859.31+/-74.02 and 753.34+/-72.95, respectively. The PED of VEGF in YLY group at each day-point was lower than that in control group (P <0.05, P <0.01, P <0.01 and P <0.01, respectively). The PED of VEGFR-2 in control group at 10th, 20th, 30th and 40th day-point was 618.63+/-59.08, 750.09+/-56.72, 684.91+/-72.86 and 644.06+/-60.25, and that in YLY group was 523.91+/-64.66, 449.03+/-46.85, 400.06+/-60.12 and 339.89+/-45.39, respectively. The PED of VEGFR-2 in YLY group at each day-point was lower than that in control group (P <0.05, P <0.01, P <0.01 and P <0.01, respectively). The PED of ES in control group at 10th, 20th, 30th and 40th day-point was 250.26+/-36.27, 298.60+/-44.41, 450.86+/-38.95 and 398.43+/-34.19, and that in YLY group was 249.57+/-40.23, 350.03+/-40.92, 499.40+/-40.29 and 497.94+/-42.76, respectively. There was no difference between the two groups at 10th day-point.The PED of ES in YLY group was higher than that in control group at 20, 30, 40 th day-point (P <0.05, P <0.01 and P <0.01, respectively) . CONCLUSION: YLY could exert the anti- tumor role by down-regulating the expression of VEGF and VEGFR-2, up-regulating the expression of ES and inhibiting the angiogenesis within tumor tissue.


Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Neoplasias/irrigação sanguínea , Neovascularização Patológica/tratamento farmacológico , Administração Oral , Animais , Medicamentos de Ervas Chinesas/administração & dosagem , Medicamentos de Ervas Chinesas/uso terapêutico , Endostatinas/metabolismo , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos BALB C , Transplante de Neoplasias , Neoplasias/tratamento farmacológico , Neoplasias/genética , Neoplasias/patologia , Neovascularização Patológica/patologia , Fator A de Crescimento do Endotélio Vascular/metabolismo , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo
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