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1.
J Am Chem Soc ; 143(24): 8993-9001, 2021 06 23.
Artigo em Inglês | MEDLINE | ID: mdl-34106720

RESUMO

The enantioselective synthesis of axially chiral biaryls by a copper-catalyzed Diels-Alder/retro-Diels-Alder reaction of 2-pyrones with alkynes is reported herein. Using electron-deficient 2-pyrones and electron-rich 1-naphthyl acetylenes as the reaction partners, a broad range of axially chiral biaryl esters are obtained in excellent yields (up to 97% yield) and enantioselectivities (up to >99% ee). DFT calculations reveal the reaction mechanism and provide insights into the origins of the stereoselectivities. The practicality and robustness of this reaction are showcased by gram-scale synthesis. The synthetic utilizations are demonstrated by the amenable transformations of the products.

2.
Org Biomol Chem ; 18(20): 3832-3837, 2020 05 27.
Artigo em Inglês | MEDLINE | ID: mdl-32396933

RESUMO

An environmentally benign electrochemically enabled site-selective functionalization of indole or aniline derivatives with hexafluoroisopropanol in the presence of tetrabutyl ammonium hexafluorophosphate as the redox catalyst and electrolyte was demonstrated in this work. Under mild electro-oxidation conditions, a series of hexafluoroisopropoxy indole and aniline derivatives with pharmacological activity were obtained. This conversion does not need transition metals and oxidants, and has good functional group tolerance. The in vitro cytotoxicity of all compounds was evaluated by the MTT assay against four human cancer cell lines. The results revealed that hexafluoroisopropoxy indoles have good antitumor activity and compound 2i increased the intracellular levels of ROS and inhibited apoptosis in HeLa cells.

3.
Anal Chim Acta ; 1096: 193-202, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31883587

RESUMO

Specific recognition of caffeic acid (CA) from Taraxacum mon-golicum Hand.-Mazz. was successfully performed using a new pH responsive magnetic molecularly imprinted polymers (pH-MMIPs) by simple surface molecular imprinting polymerization. The pH-MMIPs were prepared on the surface of the Fe3O4@SiO2@MPS particles using CA as a template, 2-(dimethylamino) ethyl methacrylate (DMA) as the pH responsive functional monomer, 4-vinylpyridine (4-VP) as an assisting functional monomer, ethylene glycol dimethyl acrylate (EGDMA) as cross-linker, 2,2'-azobisisobutyronitrile (AIBN) as initiator and methanol-H2O (1:1, v/v) as the porogen. The resultant polymers were characterized by scanning electron microscope (SEM), transmission electron microscope (TEM), dynamic light scattering (DLS), Fourier transform infrared spectroscopy (FT-IR), thermal gravimetric analysis (TGA), vibrating sample magnetometry (VSM) and x-ray diffraction (XRD). The adsorption experiments revealed that the pH-MMIPs performed high adsorption ability (11.5 mg g-1) by changing solution pH. Successful selective adsorption of CA was achieved with distribution coefficient of 0.12 and 0.21 towards ferulic acid and chlorogenic acid. Furthermore, pH-MMIPs were employed as adsorbents for extraction and enrichment of CA from Taraxacum mon-golicum Hand.-Mazz. extract. The recoveries of CA in the Taraxacum mon-golicum Hand.-Mazz. ranged from 90.47% to 98.97%. The results proved that the polymers have the potential to provide a selective recognition of CA in complex samples by simple pH regulation.


Assuntos
Ácidos Cafeicos/isolamento & purificação , Impressão Molecular/métodos , Polímeros/química , Taraxacum/química , Adsorção , Óxido Ferroso-Férrico/química , Concentração de Íons de Hidrogênio , Imãs/química , Metacrilatos/química , Polimerização , Piridinas/química , Dióxido de Silício/química
4.
Am J Physiol Cell Physiol ; 317(2): C253-C261, 2019 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-30649914

RESUMO

Myocardial ischemia-reperfusion (I/R) is a common and lethal disease that threatens people's life worldwide. The underlying mechanisms are under intensive study and yet remain unclear. Here, we explored the function of miR-322/503 in myocardial I/R injury. We used isolated rat perfused heart as an in vivo model and H9c2 cells subjected with the oxygen and glucose deprivation followed by reperfusion as in vitro model to study myocardial I/R injury. 2,3,5-Triphenyltetrazolium chloride (TTC) staining was used to measure the infarct size, and terminal deoxynucleotidyl transferase dUTP-mediated nick-end label (TUNEL) staining was used to examine apoptosis. Quantitative RT-PCR and Western blot were used to determine expression levels of miR-322/503, Smad ubiquitin regulatory factor 2 (Smurf2), enhancer of zeste homolog 2 (EZH2), p-Akt, and p-GSK3ß. Overexpression of miR-322/503 decreased infarct size, inhibited cell apoptosis, and promoted cell proliferation through upregualtion of p-Akt and p-GSK3ß. Thus the expression of miR-322/503 was reduced during I/R process. On the molecular level, miR-322/503 directly bound Smurf2 mRNA and suppressed its translation. Smurf2 ubiquitinated EZH2 and degraded EZH2, which could activate Akt/GSK3ß signaling. Our study demonstrates that miR-322/503 plays a beneficial role in myocardial I/R injury. By inhibition of Smurf2 translation, miR-322/503 induces EZH2 expression and activates Akt/GSK3ß pathway, thereby protecting cells from ischemia reperfusion injury.


Assuntos
Proteína Potenciadora do Homólogo 2 de Zeste/metabolismo , Glicogênio Sintase Quinase 3 beta/metabolismo , MicroRNAs/metabolismo , Infarto do Miocárdio/prevenção & controle , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Miócitos Cardíacos/enzimologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Animais , Apoptose , Sítios de Ligação , Hipóxia Celular , Linhagem Celular , Proliferação de Células , Modelos Animais de Doenças , Glucose/deficiência , Preparação de Coração Isolado , Masculino , MicroRNAs/genética , Infarto do Miocárdio/enzimologia , Infarto do Miocárdio/genética , Infarto do Miocárdio/patologia , Traumatismo por Reperfusão Miocárdica/enzimologia , Traumatismo por Reperfusão Miocárdica/genética , Traumatismo por Reperfusão Miocárdica/patologia , Miócitos Cardíacos/patologia , Fosforilação , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos Sprague-Dawley , Transdução de Sinais , Ubiquitina-Proteína Ligases/genética
5.
Artigo em Inglês | MEDLINE | ID: mdl-30529494

RESUMO

In this work, surface molecularly imprinted polymers (SMIPs) were prepared as a specific sorbent to remove the limonin from the lemon juice for the first time, and then the MIPs containing limonin were directly made into a water-soluble gel to treat inflammation of mice. The resulting polymers were characterized by scanning electron microscopy, transmission electron microscopy and Fourier transform infrared spectrometer spectra. And the polymerization conditions and adsorption performances of the resultant nanomaterials were further investigated in detail. Results showed that the MIPs have higher adsorption capacity (27.72 mg/g) compared with surface molecularly non-imprinted polymers (NIPs) (8.12 mg/g). The selectivity experiment indicated that the polymers had excellent selective recognition for limonin and the selectivity factors were calculated as 2.75 and 1.83 for nomilin and obakunone, respectively. The MIPs were successfully used as adsorbent for selectively removing limonin from lemon juice and the MIPs extracted almost all the limonin from lemon juice according to the HPLC results. Furthermore, the MIPs with limonin were processed into water-soluble gel, which can be used to reduce the inflammation and enhance wound healing of model mice.


Assuntos
Citrus , Sucos de Frutas e Vegetais/análise , Limoninas/isolamento & purificação , Impressão Molecular/métodos , Adsorção , Animais , Sucos de Frutas e Vegetais/normas , Limoninas/química , Limoninas/farmacologia , Camundongos , Cicatrização/efeitos dos fármacos
6.
J Sep Sci ; 41(15): 3060-3068, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29878532

RESUMO

Thermo-responsive magnetic molecularly imprinted polymers were prepared by simple surface molecular imprinting polymerization for the selective adsorption and enrichment of formononetin from Trifolium pretense by temperature regulation. Using formononetin as a template, N-isopropylacrylamide as the thermo-responsive functional monomer, and methacrylic acid as an assisting functional monomer, the polymers were synthesized on the surface of the magnetic substrate. The results show that imprinted polymers attained controlled adsorption of formononetin in response to the temperature change, with large adsorption capacity (16.43 mg/g), fast kinetics (60 min) and good selectivity at 35°C compared with that at 25 and 45°C. The selectivity experiment indicated that the materials had excellent recognition ability for formononetin and the selectivity factors were between 1.32 and 2.98 towards genistein and daidzein. The excellent linearity was attained in the range of 5-100 µg/mL, with low detection limits and low quantitation limits of 0.017 and 0.063 µg/mL, respectively. Furthermore, the thermo-responsive magnetic molecularly imprinted polymers were successfully utilized for enriching and purifying formononetin from Trifolium pretense. The analytical results indicate that the imprinted polymers are promising materials for selective identification and enrichment of formononetin in complicated herbal medicines by simple temperature-responsive regulation.


Assuntos
Isoflavonas/química , Impressão Molecular , Polímeros/química , Temperatura , Adsorção , Isoflavonas/isolamento & purificação , Fenômenos Magnéticos , Polimerização , Polímeros/síntese química , Propriedades de Superfície , Trifolium/química
7.
Food Funct ; 9(7): 3807-3814, 2018 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-29932194

RESUMO

An efficient method combined with fingerprint and chemometric analyses was developed to evaluate the quality of the traditional Chinese medicine plant Penthorum chinense Pursh. Nine samples were collected from different regions during different harvest periods, and 17 components in the form of extracts were simultaneously examined to assess quality by using high-performance liquid chromatography. The hepatoprotective effects of components were investigated by assessing the inhibition of SMMC-7721 cell growth. The results indicated that the quality control method was accurate, stable, and reliable, and the hierarchical heat-map cluster and the principle component analyses confirmed that the classification of all nine samples was consistent. Quercetin and ellagitannins including pinocembrin-7-O-[3''-O-galloyl-4'',6''-hexahydroxydiphenoyl]-ß-glucose (PGHG), thonningianin A, thonningianin B, and other flavonoids were abundant in the extracts, and significantly contributed to the hepatoprotective effects.


Assuntos
Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/farmacologia , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/farmacologia , Magnoliopsida/química , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Flavonoides/química , Humanos , Neoplasias Hepáticas/tratamento farmacológico , Neoplasias Hepáticas/fisiopatologia
8.
Ying Yong Sheng Tai Xue Bao ; 29(4): 1170-1178, 2018 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-29726226

RESUMO

To explore the changes of vegetation landscape pattern and its driving mechanism in Saihanba Nature Reserve, we analyzed vegetation type changes from 1989 to 2013 and the driving factors using random forest and Logistic regression models in conjunction with land dynamic degree indicator, based on three Landsat TM imageries obtained in 1989, 2000 and 2013. The results showed that the proportion of shrubland was always small in this area from 1989 to 2013. During 1989-2013, the proportion of shrubland rapidly decreased and plantation area significantly increased, while the area of grassland and natural secondary forest slightly changed. Key driving factors for the vegetation dynamics were dependent on vegetation type and time. The change of each vegetation type from 1989 to 2000 was significantly influenced by social factors, i.e. distance to road and total investment of afforestation. Since the implementation of the Natural Forest Protection Project and establishment of Saihanba Nature Reserve, the role of natural factors including elevation and aspect gradually became more important during the 2000-2013. The vegetation landscape dynamics were primarily determined by social activities, while the distribution patterns of vegetation types were probably controlled by natural factors in the study area.


Assuntos
Conservação dos Recursos Naturais , China , Ecossistema
9.
J Agric Food Chem ; 66(3): 653-660, 2018 Jan 24.
Artigo em Inglês | MEDLINE | ID: mdl-29260546

RESUMO

In this work, a modified pretreatment method using magnetic molecularly imprinted polymers (MMIPs) was successfully applied to study the metabolites of an important botanical with ultraperformance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). The MMIPs for glucoside-specific adsorption was used to identify metabolites of Pulsatilla chinensis in rat feces. Polymers were prepared by using Fe3O4 nanoparticles as the supporting matrix, d-glucose as fragment template, and dopamine as the functional monomer and cross-linker. Results showed that MMIPs exhibited excellent extraction performance, large adsorption capacity (5.65 mg/g), fast kinetics (60 min), and magnetic separation. Furthermore, the MMIPs coupled with UPLC-Q-TOF-MS were successfully utilized for the identification of 17 compounds including 15 metabolites from the Pulsatilla saponin metabolic pool. This study provides a reliable protocol for the separation and identification of saponin metabolites in a complex biological sample, including those from herbal medicines.


Assuntos
Fezes/química , Nanopartículas de Magnetita/química , Polímeros/química , Pulsatilla/metabolismo , Saponinas/química , Saponinas/isolamento & purificação , Extração em Fase Sólida/métodos , Animais , Cromatografia Líquida de Alta Pressão , Indóis/química , Magnetismo , Espectrometria de Massas , Impressão Molecular , Polímeros/síntese química , Pulsatilla/química , Ratos , Saponinas/metabolismo , Extração em Fase Sólida/instrumentação
10.
Cell Physiol Biochem ; 43(6): 2226-2241, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29069652

RESUMO

BACKGROUND/AIMS: The study aimed to investigate the protective effect of curcumin against oxidative stress-induced injury of Parkinson's disease (PD) through the Wnt/ß-catenin signaling pathway in rats. METHODS: The successfully established PD rat models and normal healthy rats were randomly assigned into the 6-hydroxydopamine (6-OHDA), the curcumin (Cur) and the control groups. Immunohistochemistry was used to detect the positive expression of tyrosine hydroxylase (TH), dopamine transporter (DAT) and glial fibrillary acidic protein (GFAP). Deutocerebrum primary cells were extracted and classified into the control, 6-OHDA, Cur (5, 10, 15 µmol/L), Dickkopf-1 (DKK-1) and Cur + DKK-1 groups. MTT assays, adhesion tests and TUNEL staining were used to assess cell viability, adhesion and apoptosis, respectively. Western blotting and qRT-PCR were used to examine the protein and mRNA expressions of Wnt3a and ß-catenin and the c-myc and cyclinD1 mRNA expressions. RESULTS: TH and DAT expressions in the Cur group were elevated and GFAP was reduced compared with the 6-OHDA group. Curcumin enhanced viability, survival and adhesion and attenuated apoptosis of deutocerebrum primary cells by activating the Wnt/ß-catenin signaling pathway. Higher Wnt3a and ß-catenin mRNA and protein expressions and c-myc and cyclinD1 mRNA expressions, enhanced superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) contents, decreased malondialdehyde (MDA) content and elevated mitochondrial membrane potential (∆ψm) were found in the 10 and 15 µmol/L Cur groups compared with the 6-OHDA group. However, opposite tendencies were found in the Cur + DKK-1 group compared to the 10 µmol/L Cur group. CONCLUSION: This study suggests that curcumin could protect against oxidative stress-induced injury in PD rats via the Wnt/ß-catenin signaling pathway.


Assuntos
Curcumina/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Substâncias Protetoras/farmacologia , Via de Sinalização Wnt/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Astrócitos/citologia , Astrócitos/efeitos dos fármacos , Astrócitos/metabolismo , Comportamento Animal/efeitos dos fármacos , Adesão Celular/efeitos dos fármacos , Células Cultivadas , Ciclina D1/genética , Ciclina D1/metabolismo , Modelos Animais de Doenças , Proteínas da Membrana Plasmática de Transporte de Dopamina/genética , Proteínas da Membrana Plasmática de Transporte de Dopamina/metabolismo , Proteína Glial Fibrilar Ácida/genética , Proteína Glial Fibrilar Ácida/metabolismo , Glutationa Peroxidase/metabolismo , Imuno-Histoquímica , Peptídeos e Proteínas de Sinalização Intercelular/genética , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Masculino , Malondialdeído/metabolismo , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Oxidopamina/farmacologia , Doença de Parkinson/patologia , Doença de Parkinson/veterinária , Proteínas Proto-Oncogênicas c-myc/genética , Proteínas Proto-Oncogênicas c-myc/metabolismo , Ratos , Ratos Sprague-Dawley , Superóxido Dismutase/metabolismo , Tirosina 3-Mono-Oxigenase/genética , Tirosina 3-Mono-Oxigenase/metabolismo , Proteína Wnt3/genética , Proteína Wnt3/metabolismo , beta Catenina/genética , beta Catenina/metabolismo
11.
Biomed Chromatogr ; 31(5)2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-27761923

RESUMO

Pulsatilla chinensis (Bunge) Regel is commonly used in Asia, and anemoside B4 (AB4) is its major saponin, with diverse pharmaceutical effects. Previous studies showed that intestinal flora plays an important role in the metabolism of herbs administered orally. In this study, the metabolic profile of AB4 with microflora in rat small and large intestines in vitro was investigated. Gut microflora was collected from different intestinal segments and anaerobically incubated with AB4 at 37°C for 24, 48, 72 and 96 h, respectively. A total of 10 metabolites were detected and identified by ultra- performance liquid chromatography/quadrupole time-of-flight mass spectrometry, involving the products of oxygenation and deglycosylation reactions. Gut microflora in the large intestine generated more comprehensive metabolic pathways, which appears to be attributable to the wider range of bacterial types and numbers of bacteria. Human cancer cell lines SMMC-7721, Hela and MCF-7 were treated with metabolite pools by MTT assay, together with M6 as the greatest deglycosylation product. As a result, M6 exhibited a reduction in cell viability of SMMC-7721 with an IC50 value of 22.28 ± 1.26 µg/mL. The present study provided scientific evidence for AB4 metabolism in small and large intestines, which is helpful to reveal the active forms of AB4 in vivo.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Intestino Grosso/microbiologia , Intestino Delgado/microbiologia , Saponinas/farmacocinética , Espectrometria de Massas em Tandem/métodos , Animais , Biotransformação , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Microbioma Gastrointestinal , Humanos , Intestino Grosso/efeitos dos fármacos , Intestino Grosso/metabolismo , Intestino Delgado/efeitos dos fármacos , Intestino Delgado/metabolismo , Masculino , Estrutura Molecular , Ratos Sprague-Dawley , Saponinas/química , Saponinas/metabolismo , Saponinas/farmacologia
12.
Am J Chin Med ; 44(6): 1221-1236, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27744729

RESUMO

Drug-induced liver injury (DILI) is the most common cause of acute liver failure. Disruption of the Th17/Treg balance can lead to hepatic inflammation, which causes the main symptoms of DILI. Here we investigate the protective mechanisms of (-)-Epigallocatechin-3-gallate (EGCG) on triptolide (TP)-induced DILI that shows the Th17/Treg imbalance. Pretreatment with EGCG (5[Formula: see text]mg/kg) for 10 days before TP (0.5[Formula: see text]mg/kg) administration in mice significantly reduced the increased alanine aminotransferase (ALT) level ([Formula: see text]) induced by TP treatment. The hepatic histology analysis further proved that EGCG protected mice from TP-induced liver injury. The imbalance of Th17/Treg was induced by TP treatment, as shown by the upregulation of TLR4 and downregulation of Tim3 expression. EGCG pretreatment can maintain the expression of TLR4 and Tim3 at normal levels to restore the Th17/Treg imbalance. In addition, EGCG can block the TP-induced expression of the downstream targets of TLR4, including MyD88, NF[Formula: see text]B, and retinoid related orphan receptor (ROR-[Formula: see text]t), while EGCG can restore the TP inhibition of forkhead/winged-helix family transcriptional repressor p3 (FoxP3) that is the downstream target of Tim3. Consequently, EGCG pretreatment can effectively inhibit the Th17-related pro-inflammatory cytokine (e.g. IL-17 and IL-6) upregulation induced by TP treatment. However, TP inhibition of Treg-related anti-inflammatory cytokine IL-10 production was restored by EGCG pretreatment. Taken together, these results suggest that EGCG possesses significant protective properties against TP-induced hepatic inflammatory injury, and that these properties are carried out via the restoration of the Th17/Treg imbalance by the inhibition of the TLR4 signaling pathway and the enhanced activation of the Tim3 signaling pathway.


Assuntos
Catequina/análogos & derivados , Doença Hepática Induzida por Substâncias e Drogas/imunologia , Doença Hepática Induzida por Substâncias e Drogas/prevenção & controle , Diterpenos/efeitos adversos , Imunossupressores/efeitos adversos , Fenantrenos/efeitos adversos , Fitoterapia , Linfócitos T Reguladores/imunologia , Células Th17/imunologia , Animais , Catequina/administração & dosagem , Catequina/farmacologia , Doença Hepática Induzida por Substâncias e Drogas/etiologia , Citocinas/metabolismo , Modelos Animais de Doenças , Compostos de Epóxi/efeitos adversos , Feminino , Receptor Celular 2 do Vírus da Hepatite A , Mediadores da Inflamação/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Transdução de Sinais/efeitos dos fármacos , Receptor 4 Toll-Like , Regulação para Cima/efeitos dos fármacos
13.
Mitochondrial DNA A DNA Mapp Seq Anal ; 27(3): 1610-1, 2016 05.
Artigo em Inglês | MEDLINE | ID: mdl-25366269

RESUMO

The Rattus norvegicus SILN strain is a common used model for nervous system disorder disease study. We sequenced this R. norvegicus strain SILN mitochondrial genome for the first time (GenBank Accession No. KM114606). Its mitogenome was 16,311 bp and coding 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNA genes.


Assuntos
Doenças do Sistema Nervoso Central/genética , Genoma Mitocondrial , Animais , Composição de Bases , Doenças do Sistema Nervoso Central/patologia , Códon de Iniciação , Códon de Terminação , DNA Mitocondrial/química , DNA Mitocondrial/genética , Bases de Dados Genéticas , Modelos Animais de Doenças , Fases de Leitura Aberta/genética , RNA Ribossômico/química , RNA Ribossômico/genética , RNA de Transferência/química , RNA de Transferência/genética , Ratos
14.
Int J Clin Exp Med ; 8(9): 15846-51, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26629086

RESUMO

BACKGROUND AND AIMS: Endoscopic Interventional Treatment is of little trauma and less complications in the treatment of esophageal tumor and leads to faster recovery and fewer days of hospitalization. This study was aimed to investigate the safety and efficacy of endoscopic interventional therapy for huge esophageal tumor arising in the muscularis propria. METHODS: The patient was treated by submucosal tunneling endoscopic resection (STER). RESULTS: The huge esophageal tumor was resected completely by STER technique, with little trauma and less complications. The size of the resected tumor was 5.5×3.5×3.0 cm. CONCLUSION: Submucosal tunneling endoscopic resection is a safe and efficient technique for treating Huge Esophageal Tumor originating from muscularis propria layer.

15.
World J Gastroenterol ; 21(36): 10367-74, 2015 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-26420963

RESUMO

AIM: To evaluate the efficacy of ursodeoxycholic acid (UDCA) as a chemotherapeutic agent for the treatment of hepatocellular carcinoma (HCC). METHODS: BALB/c nude mice were randomized into four groups 24 h before subcutaneous injection of hepatocarcinoma BEL7402 cells suspended in phosphate buffered saline (PBS) into the right flank. The control group (n = 10) was fed a standard diet while treatment groups (n = 10 each) were fed a standard daily diet supplemented with different concentrations of UDCA (30, 50 and 70 mg/kg per day) for 21 d. Tumor growth was measured once each week, and tumor volume (V) was calculated with the following equation: V = (L × W(2)) × 0.52, where L is the length and W is the width of the xenograft. After 21 d, mice were killed under ether anesthesia, and tumors were excised and weighed. Apoptosis was evaluated through detection of DNA fragmentation with gel electrophoresis and the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay. Western blot analysis was performed to determine the expression of apoptosis-related proteins BAX, BCL2, APAF1, cleaved caspase-9, and cleaved caspase-3. RESULTS: UDCA suppressed tumor growth relative to controls. The mean tumor volumes were the following: control, 1090 ± 89 mm(3); 30 mg/kg per day, 612 ± 46 mm(3); 50 mg/kg per day, 563 ± 38 mm(3); and 70 mg/kg per day, 221 ± 26 mm(3). Decreased tumor volumes reached statistical significance relative to control xenografts (30 mg/kg per day, P < 0.05; 50 mg/kg per day, P < 0.05; 70 mg/kg per day, P < 0.01). Increasing concentrations of UDCA led to increased DNA fragmentation observed on gel electrophoresis and in the TUNEL assay (control, 1.6% ± 0.3%; 30 mg/kg per day, 2.9% ± 0.5%; 50 mg/kg per day, 3.15% ± 0.7%, and 70 mg/kg per day, 4.86% ± 0.9%). Western blot analysis revealed increased expression of BAX, APAF1, cleaved-caspase-9 and cleaved-caspase-3 proteins, which induce apoptosis, but decreased expression of BCL2 protein, which is an inhibitor of apoptosis, following administration of UDCA. CONCLUSION: UDCA suppresses growth of BEL7402 hepatocellular carcinoma cells in vivo, in part through apoptosis induction, and is thus a candidate for therapeutic treatment of HCC.


Assuntos
Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Carcinoma Hepatocelular/tratamento farmacológico , Neoplasias Hepáticas/tratamento farmacológico , Ácido Ursodesoxicólico/farmacologia , Animais , Proteínas Reguladoras de Apoptose/metabolismo , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral , Relação Dose-Resposta a Droga , Humanos , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patologia , Masculino , Camundongos Endogâmicos BALB C , Camundongos Nus , Pessoa de Meia-Idade , Fatores de Tempo , Carga Tumoral/efeitos dos fármacos , Ensaios Antitumorais Modelo de Xenoenxerto
16.
J Virol ; 89(20): 10532-47, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26269185

RESUMO

UNLABELLED: Despite the identification of horseshoe bats as the reservoir of severe acute respiratory syndrome (SARS)-related coronaviruses (SARSr-CoVs), the origin of SARS-CoV ORF8, which contains the 29-nucleotide signature deletion among human strains, remains obscure. Although two SARS-related Rhinolophus sinicus bat CoVs (SARSr-Rs-BatCoVs) previously detected in Chinese horseshoe bats (Rhinolophus sinicus) in Yunnan, RsSHC014 and Rs3367, possessed 95% genome identities to human and civet SARSr-CoVs, their ORF8 protein exhibited only 32.2 to 33% amino acid identities to that of human/civet SARSr-CoVs. To elucidate the origin of SARS-CoV ORF8, we sampled 348 bats of various species in Yunnan, among which diverse alphacoronaviruses and betacoronaviruses, including potentially novel CoVs, were identified, with some showing potential interspecies transmission. The genomes of two betacoronaviruses, SARSr-Rf-BatCoV YNLF_31C and YNLF_34C, from greater horseshoe bats (Rhinolophus ferrumequinum), possessed 93% nucleotide identities to human/civet SARSr-CoV genomes. Although these two betacoronaviruses displayed lower similarities than SARSr-Rs-BatCoV RsSHC014 and Rs3367 in S protein to civet SARSr-CoVs, their ORF8 proteins demonstrated exceptionally high (80.4 to 81.3%) amino acid identities to that of human/civet SARSr-CoVs, compared to SARSr-BatCoVs from other horseshoe bats (23.2 to 37.3%). Potential recombination events were identified around ORF8 between SARSr-Rf-BatCoVs and SARSr-Rs-BatCoVs, leading to the generation of civet SARSr-CoVs. The expression of ORF8 subgenomic mRNA suggested that the ORF8 protein may be functional in SARSr-Rf-BatCoVs. The high Ka/Ks ratio among human SARS-CoVs compared to that among SARSr-BatCoVs supported that ORF8 is under strong positive selection during animal-to-human transmission. Molecular clock analysis using ORF1ab showed that SARSr-Rf-BatCoV YNLF_31C and YNLF_34C diverged from civet/human SARSr-CoVs in approximately 1990. SARS-CoV ORF8 originated from SARSr-CoVs of greater horseshoe bats through recombination, which may be important for animal-to-human transmission. IMPORTANCE: Although horseshoe bats are the primary reservoir of SARS-related coronaviruses (SARSr-CoVs), it is still unclear how these bat viruses have evolved to cross the species barrier to infect civets and humans. Most human SARS-CoV epidemic strains contain a signature 29-nucleotide deletion in ORF8, compared to civet SARSr-CoVs, suggesting that ORF8 may be important for interspecies transmission. However, the origin of SARS-CoV ORF8 remains obscure. In particular, SARSr-Rs-BatCoVs from Chinese horseshoe bats (Rhinolophus sinicus) exhibited <40% amino acid identities to human/civet SARS-CoV in the ORF8 protein. We detected diverse alphacoronaviruses and betacoronaviruses among various bat species in Yunnan, China, including two SARSr-Rf-BatCoVs from greater horseshoe bats that possessed ORF8 proteins with exceptionally high amino acid identities to that of human/civet SARSr-CoVs. We demonstrated recombination events around ORF8 between SARSr-Rf-BatCoVs and SARSr-Rs-BatCoVs, leading to the generation of civet SARSr-CoVs. Our findings offer insight into the evolutionary origin of SARS-CoV ORF8 protein, which was likely acquired from SARSr-CoVs of greater horseshoe bats through recombination.


Assuntos
Infecções por Coronavirus/veterinária , Genoma Viral , RNA Viral/genética , Recombinação Genética , Vírus da SARS/genética , Proteínas da Matriz Viral/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , China , Quirópteros/virologia , Infecções por Coronavirus/genética , Infecções por Coronavirus/transmissão , Infecções por Coronavirus/virologia , Evolução Molecular , Expressão Gênica , Humanos , Dados de Sequência Molecular , Filogenia , Filogeografia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Viral/metabolismo , Vírus da SARS/classificação , Vírus da SARS/metabolismo , Alinhamento de Sequência , Homologia de Sequência do Ácido Nucleico , Síndrome Respiratória Aguda Grave/genética , Síndrome Respiratória Aguda Grave/metabolismo , Síndrome Respiratória Aguda Grave/transmissão , Síndrome Respiratória Aguda Grave/virologia , Proteínas da Matriz Viral/metabolismo , Viverridae/virologia
17.
BMC Res Notes ; 8: 255, 2015 Jun 23.
Artigo em Inglês | MEDLINE | ID: mdl-26100251

RESUMO

BACKGROUND: There have been four strains on Manzanilla virus (MANV) identified to date. Here, we identify a novel MANV strain (DHL10M107) isolated from Culex tritaeniorhynchus Giles mosquitoes from Ruili city, Dehong prefecture, Yunnan Province, in the People's Republic of China. RESULTS: The DHL10M107 L, M and S genes were sequenced at the nucleotide and deduced amino acid levels. The L, M and S gene sequences of DHL10M107 clustered with the MANV strains VN04-2108, TRVL3587, SA An 4165, and AV 782. DHL10M107 was most closely related to VN04-2108. Nucleotide homology ranged between 96 and 99% between DHL10M107 and VN04-2108. In terms of amino acid homology, all of the amino acid differences were in the L (96.3% homologous) and M (97.7% homologous) fragments. CONCLUSIONS: DHL10M107 is likely a MANV isolated from mosquitos in the Yunnan Province. This is the first reported isolation of MANV in mainland China.


Assuntos
Culex/virologia , Genoma Viral , Orthobunyavirus/genética , Filogenia , Proteínas Virais/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , China , Orthobunyavirus/classificação , Orthobunyavirus/isolamento & purificação , Homologia de Sequência de Aminoácidos
18.
Int J Clin Exp Pathol ; 8(10): 12093-100, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26722394

RESUMO

OBJECTIVE: This study aims to explore the protection effect of bone marrow mesenchymal stem cells (BMSCs) on PC12 cells apoptosis mediated by transient axonal glycoprotein 1 (TAG1). METHODS: PC12 cells were divided into control group, Aß25-35 group and BMSCs + Aß25-35 group. The effects of BMSCs on PC12 cells treated by Aß25-35 were detected using MTT, Hoechst 33258 and Annexin V-FITC/PI staining methods. The expression levels of TAG1, ß-amyloid precursor protein (APP), AICD and p53 were determined by RT-PCR and Western blotting methods. The expression levels of Bax and Bcl-2 were determined by Western blotting method. The activity of Caspase 3 was detected by spectrophotometric method. RESULTS: MTT results showed that cell activity decreased after the treatment of 20 µM Aß25-35 for 48 h (P<0.01) while it increased in BMSCs + Aß25-35 group (P<0.01). Hoechst 33258 and Annexin V-FITC/PI staining results showed that Aß25-35 could induce the apoptosis of PC12 cells while the apoptosis of PC12 cells was inhibited in BMSCs + Aß25-35 group. RT-PCR and Western blotting methods showed that 20 µM Aß25-35 could increase the expression levels of TAG1, APP, AICD and p53 (P<0.01) while they decreased in BMSCs + Aß25-35 group (P<0.01). 20 µM Aß25-35 could increase the expression levels of Bax and decrease the expression levels of Bcl-2 (P<0.01), while the expression levels of Bax decreased and the expression levels of Bcl-2 increase in BMSCs + Aß25-35 group (P<0.01). 20 µM Aß25-35 could enhance Caspase 3 activity while it decreased in BMSCs + Aß25-35 group (P<0.01). Conclusions BMSCs with Aß25-35 could inhibit the apoptosis of PC12 cells, which maybe related with TAG1/APP/AICD signal pathway.


Assuntos
Peptídeos beta-Amiloides/toxicidade , Apoptose/fisiologia , Contactina 2/metabolismo , Células-Tronco Mesenquimais/metabolismo , Neurônios/efeitos dos fármacos , Animais , Western Blotting , Células da Medula Óssea/metabolismo , Técnicas de Cocultura , Citometria de Fluxo , Neurônios/metabolismo , Neurônios/patologia , Células PC12 , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real , Transdução de Sinais/efeitos dos fármacos
19.
Cell Transplant ; 23 Suppl 1: S113-22, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25385295

RESUMO

Multiple sclerosis (MS) is a complex disease of neurological disability, affecting more than 300 out of every 1 million people in the world. The purpose of the study was to evaluate the therapeutic effects of human umbilical cord-derived mesenchymal stem cell (hUC-MSC) transplantation in MS patients. Twenty-three patients were enrolled in this study, and 13 of them were given hUC-MSC therapy at the same time as anti-inflammatory treatment, whereas the control patients received the anti-inflammatory treatment only. Treatment schedule included 1,000 mg/kg of methylprednisolone intravenously (IV) daily for 3 days and then 500 mg/kg for 2 days, followed by oral prednisone 1 mg/kg/day for 10 days. The dosage of prednisone was then reduced by 5 mg every 2 weeks until reaching a 5-mg/day maintenance dosage. Intravenous infusion of hUC-MSCs was applied three times in a 6-week period for each patient. The overall symptoms of the hUC-MSC-treated patients improved compared to patients in the control group. Both the EDSS scores and relapse occurrence were significantly lower than those of the control patients. Inflammatory cytokines were assessed, and the data demonstrated a shift from Th1 to Th2 immunity in hUC-MSC-treated patients. Our data demonstrated a high potential for hUC-MSC treatment of MS. This manuscript is published as part of the International Association of Neurorestoratology (IANR) special issue of Cell Transplantation.


Assuntos
Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/citologia , Esclerose Múltipla/fisiopatologia , Cordão Umbilical/citologia , Adulto , Citocinas/sangue , Feminino , Humanos , Masculino , Esclerose Múltipla/sangue , Esclerose Múltipla/terapia , Recidiva
20.
Biomed Res Int ; 2014: 909657, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25276829

RESUMO

Parkinson's disease (PD) is a neurodegenerative movement disorder that is characterized by the progressive degeneration of the dopaminergic (DA) pathway. Mesenchymal stem cells derived from human umbilical cord (hUC-MSCs) have great potential for developing a therapeutic agent as such. HGF is a multifunctional mediator originally identified in hepatocytes and has recently been reported to possess various neuroprotective properties. This study was designed to investigate the protective effect of hUC-MSCs infected by an adenovirus carrying the HGF gene on the PD cell model induced by MPP+ on human bone marrow neuroblastoma cells. Our results provide evidence that the cultural supernatant from hUC-MSCs expressing HGF could promote regeneration of damaged PD cells at higher efficacy than the supernatant from hUC-MSCs alone. And intracellular free Ca(2+) obviously decreased after treatment with cultural supernatant from hUC-MSCs expressing HGF, while the expression of CaBP-D28k, an intracellular calcium binding protein, increased. Therefore our study clearly demonstrated that cultural supernatant of MSC overexpressing HGF was capable of eliciting regeneration of damaged PD model cells. This effect was probably achieved through the regulation of intracellular Ca(2+) levels by modulating of CaBP-D28k expression.


Assuntos
Adenoviridae/metabolismo , Fator de Crescimento de Hepatócito/metabolismo , Células-Tronco Mesenquimais/metabolismo , Regeneração Nervosa , Doença de Parkinson/patologia , Cordão Umbilical/citologia , Calbindina 1/metabolismo , Cálcio/metabolismo , Linhagem Celular , Separação Celular , Sobrevivência Celular , Meios de Cultivo Condicionados/farmacologia , Ensaio de Imunoadsorção Enzimática , Humanos , Espaço Intracelular/metabolismo , Células-Tronco Mesenquimais/citologia , Modelos Biológicos , Neurônios , Transdução Genética
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