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1.
Anal Chem ; 2020 Mar 26.
Artigo em Inglês | MEDLINE | ID: mdl-32212632

RESUMO

Glycan modification prompts important concerns about the quality control of biopharmaceutical production. Conbercept is a multi-glycosylated recombinant fusion protein drug approved for the treatment of age-related macular degeneration (AMD). With 14 N-glycosites in the molecule and 7 N-glycosites in the monomer, the charge isomer separation and characterization of conbercept pose great challenges due to its enormous heterogeneities. The batch-to-batch stability on the charge isomer distribution and the possible causation of the pattern necessitate the development of effective analytical approaches. Here, the immobilized pH gradient (IPG)-based two-dimensional gel electrophoresis (2-DE) approach was first optimized to achieve high-resolution, high-reproducible separation and preparation of charge isomers. Then, combined with the quantitative analysis strategy of site-specific N-glycan heterogeneity based on the diagnostic MS2 ion (peptides+GlcNAc, Y1 ions) of glycopeptides, an integrated approach for the quantitation of site-specific N-glycan heterogeneities among charge isomers was established. Finally, the quantitation of site-specific N-glycoforms in each of the 2-DE resolved spots were performed, and the results showed that the sialylaion tends to increase for gel spots located in the acidic regions. This study provides an effective approach to separate the charge isomers of the heavily glycosylated protein drugs, and to quantitatively explore the site-specific N-glycans dynamics along with the different charge isomers.

2.
Int J Biol Macromol ; 149: 1213-1221, 2020 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-32035955

RESUMO

Lectins are the primary recognition macromolecules for various types of fucosylation, a common eukaryotic post-translational modification. In this study, we report the heterologous expression and molecular binding properties of a fucose-specific lectin, AofleA, isolated from Arthrobotrys oligospora. This is the first reported fucose-specific lectin found in nematophagous fungi. The recombinant AofleA (r-AofleA) was expressed in Escherichia coli with high efficiency, yielding at least 500 mg of soluble and functional r-AofleA per liter of broth. Using hemagglutination inhibition assay and glycan microarray analysis, r-AofleA was found to be broadly specific for fucosylated glycans or oligosaccharides including Fucα(1-2), Fucα(1-3), Fucα(1-4) and Fucα(1-6) linkages, similar to Aleuria aurantia lectin (AAL). Frontal affinity chromatography showed that r-AofleA has high affinity towards PA-L-fucose with an average Kd value of 15 nM. These findings provide a basis for improved understanding of the structure and functions of AofleA during recognition and capture of prey nematodes by nematophagous fungus A. oligospora.

3.
Zhongguo Fei Ai Za Zhi ; 23(3): 156-161, 2020 Mar 20.
Artigo em Chinês | MEDLINE | ID: mdl-32102132

RESUMO

BACKGROUND: Thymectomy plays an important role in patients with myasthenia gravis (MG) and video-assisted thoracoscopic (VATS) extended thymectomy has been widely used in recent years. The traditional thoracoscopic procedure requires three intercostal incisions. Our study improved the surgical approach and operation method by using single-utility port thoracoscopy. The aim of this study was to evaluate the effect of single-utility port VATS extended thymectomy for MG. METHODS: A retrospective analysis was performed in 45 patients with MG who underwent single-utility port VATS extended thymectomy in Beijing Hospital from July 2017 to December 2018. The safety and effectiveness of this surgical approach were summarized. RESULTS: All operations were accomplished successfully, without conversion to thoracotomy, requirement of additional incisions or postoperative death. The mean operation time was (141.3±39.2) min; the mean intraoperative blood loss was (64.2±45.5) mL; the median thoracic drainage duration was 3 days (range 2 days to 8 days); the mean pleural drainage was (890.4±439.1) mL, and the median postoperative hospital stay was 6 days (range 3 days to 91 days). There were 13 cases (28.9%) with postoperative complications, including 5 cases (11.1%) with myasthenia crisis, 6 cases (13.3%) with pulmonary complications, 4 cases (8.9%) with poor incision healing, 4 cases (8.9%) with atrial fibrillation, and 1 case (2.2%) with delayed pericardial hemorrhage. The median follow-up time was 18.5 months (range 12.5 months to 29.2 months). According to the effect of 1 year after surgery, 1 case (2.2%) showed pharmacologic remission; 18 cases (40.0%) showed minimal manifestations; 23 cases (51.1%) got improvement; 1 case (2.2%) showed no changs and 2 cases (4.4%) were worse. CONCLUSIONS: Single-utility port VATS extended thymectomy is a safe and feasible minimally invasive procedure for MG. During the perioperative period, special attention should be paid to prevent myasthenic crisis, pulmonary complications, and incision complications.

4.
FEBS Lett ; 2020 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-32027024

RESUMO

Peroxiredoxins (Prxs) are thiol-specific antioxidant proteins commonly found in organisms that protect cells from the damage of reactive oxygen species produced by metabolism and that participate in cell signaling. The Prx from the bacterium Akkermansia muciniphila (AmPrx) is a typical 2-Cys Prx characterized by two conserved cysteines: Cys49 and Cys183. Here, we verified the peroxidase activity of AmPrx and determined its crystal structure in reduced form, which is a doughnut-shaped decamer composed of five dimers. Particularly, a distinct loop between the α4 helix and ß6 strand is involved in the decameric interaction. Deletion of this loop destroys the decameric structure and significantly decreases the peroxidase activity of AmPrx. Our findings reveal a novel regulatory mechanism of typical 2-Cys Prx, in which the α4-ß6 loop affects the assembly of Prx and, therefore, regulates its peroxidase activity.

5.
Plant Mol Biol ; 102(4-5): 537-551, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31916084

RESUMO

KEY MESSAGE: Silencing of SlCAND1 expression resulted in dwarfish, loss of apical dominance, early flowering, suppression of seed germination, and abnormal root architecture in tomato Cullin-RING E3 ligases (CRLs)-dependent ubiquitin proteasome system mediates degradation of numerous proteins that controls a wide range of developmental and physiological processes in eukaryotes. Cullin-associated Nedd8-dissociated protein 1 (CAND1) acts as an exchange factor allowing substrate recognition part exchange and plays a vital role in reactivating CRLs. The present study reports on the identification of SlCAND1, the only one CAND gene in tomato. SlCAND1 expression is ubiquitous and positively regulated by multiple plant hormones. Silencing of SlCAND1 expression using RNAi strategy resulted in a pleiotropic and gibberellin/auxin-associated phenotypes, including dwarf plant with reduced internode length, loss of apical dominance, early flowering, low seed germination percentage, delayed seed germination speed, short primary root, and increased lateral root proliferation and elongation. Moreover, application of exogenous GA3 or IAA could partly rescue some SlCAND1-silenced phenotypes, and the expression levels of gibberellin/auxin-related genes were altered in SlCAND1-RNAi lines. These facts revealed that SlCAND1 is required for gibberellin/auxin-associated regulatory network in tomato. Although SlCAND1 is crucial for multiple developmental processes during vegetative growth stage, SlCAND1-RNAi lines didn't exhibit visible effect on fruit development and ripening. Meanwhile, we discussed that multiple physiological functions of SlCAND1 in tomato are different to previous report of its ortholog in Arabidopsis. Our study adds a new perspective on the functional roles of CAND1 in plants, and strongly supports the hypothesis that CAND1 and its regulated ubiquitin proteasome system are pivotal for plant vegetative growth but possibly have different roles in diverse plant species.


Assuntos
Flores/fisiologia , Germinação , Lycopersicon esculentum/fisiologia , Proteínas de Plantas/fisiologia , Raízes de Plantas/fisiologia , Arabidopsis/genética , Proteínas Culina , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Giberelinas/metabolismo , Lycopersicon esculentum/genética , Fenótipo , Reguladores de Crescimento de Planta/fisiologia , Proteínas de Plantas/genética , Interferência de RNA , Sementes/fisiologia , Fatores de Transcrição/genética , Fatores de Transcrição/fisiologia
6.
ACS Appl Mater Interfaces ; 12(4): 4333-4342, 2020 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-31935068

RESUMO

The abuse of antibiotics resulted in the emergence of antibiotics-resistant bacteria, which has raised a great social concern together with the impetus to develop effective antibacterial materials. Herein, the synthesis of biocompatible enzyme-responsive Ag nanoparticle assemblies (ANAs) and their application in the high-efficiency targeted antimicrobial treatment of methicillin-resistant Staphylococcus aureus (MRSA) have been demonstrated. The ANAs could collapse and undergo stable/collapsed transition on approaching MRSA because of the serine protease-like B enzyme proteins (SplB)-triggered decomposition of the branched copolymers which have been employed as the macrotemplate in the synthesis of responsive ANAs. This transition contributed greatly to the high targeting affinity and efficiency of ANAs to MRSA. The minimum inhibitory concentration and minimum bactericidal concentration against MRSA were 2.0 and 32.0 µg mL-1, respectively. Skin wound healing experiments confirmed that the responsive ANAs could serve as an effective wound dressing to accelerate the healing of MRSA infection.

7.
Zhongguo Fei Ai Za Zhi ; 22(7): 427-432, 2019 Jul 20.
Artigo em Chinês | MEDLINE | ID: mdl-31315781

RESUMO

BACKGROUND: DNA polymerase ß is one of the key enzymes for DNA repair and it was reported that about 30 percent of different types of cancers carried mutations in its coding gene Polb. However, it is still controversial whether it is true or false because of the small sample size in these studies. In current study, we performed genetic screening of promoter and coding regions of Polb gene in 69 Chinese lung cancer patients using Sanger sequencing method, so as to elucidate real mutation frequency of Polb mutations in Chinese Han population. METHODS: Salting out extraction method was used to get the genome DNAs from tumor and normal matched tissues of 69 lung cancer patients. The promoter and 14 coding regions of Polb gene were then amplified using these DNAs as the template. After purification, amplicons were sequenced and aligned to the wild type Polb gene in NCBI database, in order to find out the mutated sites of Polb gene in Chinese lung cancer patients. RESULTS: In this study, we totally found only 5 mutated sites in Polb gene. In detail, 3 mutations (-196G>T, -188_-187insCGCCC, -168C>A) were located in the promoter region; 2 mutations (587C>G, 612A>T) were found in coding regions. Specially, mutations of -188_-187insCGCCC and 587C>G (resulting to the amino acid substitution of Thr to Ser at position 196) had never been reported by other groups before. However, all these 5 mutated sites could be detected in both tumor and matched normal tissues, which inferred that they are not lung tumor specific mutations. CONCLUSIONS: No lung tumor specific mutations of Polb gene could be found in Chinese lung cancer patients and Polb gene mutation might not be a molecular marker for Chinese lung cancer patients.


Assuntos
Análise Mutacional de DNA , DNA Polimerase beta/genética , Testes Genéticos , Neoplasias Pulmonares/enzimologia , Neoplasias Pulmonares/genética , Idoso , Idoso de 80 Anos ou mais , Sequência de Bases , Detecção Precoce de Câncer , Éxons/genética , Feminino , Humanos , Neoplasias Pulmonares/diagnóstico , Masculino , Pessoa de Meia-Idade
8.
Biochem Biophys Res Commun ; 517(1): 29-35, 2019 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-31345574

RESUMO

In this paper, we characterized Am2136 as a ß-N-acetylhexosaminidase from Akkermansia muciniphila to perform the biochemical characteristics and the crystal structure of selenomethionine-labeled Am2136 with GlcNAc complex. Crystallographic evidence suggests that an oxazolinium ion was formed intermediately by the 2-acetamido group during the substrate-assisted catalytic procedure. Structural and kinetic analysis of native Am2136 and D412A, E413A mutants were investigated and the results revealed substantial difference. The Kcat/Km value of D412A was decreased 4297-fold compared to native Am2136 revealed that mutation of Asp-412 results in preventing the 2-acetamido substituent from providing anchimeric assistance and thus reducing the catalytic efficiency. Moreover, Am2136 has a wide dependence on pH and temperature, while sensitive to divalent metal ions such as Ca2+ and Mn2+. These biochemical and crystallographic results provide evidences that Asp-412 residue assists to orient the 2-acetamido group for catalysis. Based on crystallographic evidence and sequence alignment with other GH family 20 enzymes, Asp-412 residue is possibly fundamental for Am2136 during substrate-assisted catalysis.

9.
Opt Express ; 27(13): 18787-18793, 2019 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-31252815

RESUMO

Continuous real-time measurements are demonstrated from a 200Gb/s format configurable CFP transceiver that uses dual-polarization probabilistic-shaped 16QAM (DP-PS16QAM) modulation. Placed in a 50GHz coherent DWDM transmission system, DP-PS16QAM achieves a back-to-back 1.8dB OSNR gain over uniform DP-16QAM. It also transports over 1940km with EDFA-only amplification, thus doubling propagation distance of uniform DP-16QAM. Furthermore, a 1Tb/s super-channel consisting of five 200Gb/s DP-PS16QAM sub-carriers is placed in a 200GHz grid, and it achieves over 1600km transmission and 5b/s/Hz SE with a raw SE at 6.86b/s/Hz.

10.
Bioresour Technol ; 287: 121414, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31078813

RESUMO

An increase in the total lipid content in algal cells under stress conditions is often accompanied by an increase in reactive oxygen species (ROS). However, the link between these two events is unclear. In this study, the regulatory mechanism of ROS formation on lipid accumulation in C. pyrenoidosa was investigated using a Fenton-like reaction. A high Spearman correlation coefficient of 0.901 was obtained between the Hydroxyl radical (OH) level and lipid content. Importantly, the increase in the total lipid content was clearly coupled with a significant increase in the intracellular OH concentration rather than increases in the H2O2 and O2- concentrations. Transcriptome data confirms that most of the differential expression genes (DEGs) involved in fatty acid and glycerolipid biosynthesis were up-regulated by the increased OH under stress conditions. These results reveal that lipid accumulation in algal cells was promoted by OH.


Assuntos
Chlorella/metabolismo , Lipídeos/biossíntese , Espécies Reativas de Oxigênio/metabolismo , Peróxido de Hidrogênio/metabolismo , Radical Hidroxila/metabolismo , Espaço Intracelular/metabolismo
11.
Mol Med Rep ; 19(5): 3548-3554, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30864692

RESUMO

Previously, we demonstrated that Rhizoma Paridis saponins (RPS), the major active component of Rhizoma Paridis, may exhibit hepatoprotective effects. The present study aimed to identify the potential mechanism of RPS on hepatic injury and improvement in hepatic fibrosis (HF). A HF model was created in Sprague­Dawley rats by administration of carbon tetrachloride. RPS was administered for treatment following creation of the HF model. The protein and mRNA expression of vascular endothelial growth factor (VEGF), platelet­derived growth factor (PDGF), extracellular signal­regulated kinase (ERK)1/2 and α­smooth muscle actin (SMA) was detected by reverse transcription quantitative polymerase chain reaction and western blot analysis. RPS was demonstrated to improve hepatic inflammation and decrease HF severity according to hematoxylin and eosin and Masson trichrome staining. Following RPS treatment, the level of alanine aminotransferase, aspartate aminotransferase and malondialdehyde, and expression levels of the mRNA and protein of VEGF, ERK1/2, PDGF and α­SMA in the model group was decreased. By contrast, the content of glutathione­PX and superoxide dismutase was increased. These data suggest that RPS may treat HF primarily through downregulation of the expression levels of the mRNA and phosphorylated VEGF, ERK1/2, PDGF and α­SMA proteins.


Assuntos
Regulação da Expressão Gênica/efeitos dos fármacos , Neovascularização Fisiológica/efeitos dos fármacos , Neovascularização Fisiológica/genética , Rizoma/química , Saponinas/farmacologia , Actinas/genética , Actinas/metabolismo , Animais , Biomarcadores , Biópsia , Cromatografia Líquida de Alta Pressão , Imuno-Histoquímica , Cirrose Hepática/tratamento farmacológico , Cirrose Hepática/genética , Cirrose Hepática/metabolismo , Cirrose Hepática/patologia , Masculino , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Fator de Crescimento Derivado de Plaquetas/genética , Fator de Crescimento Derivado de Plaquetas/metabolismo , Ratos , Saponinas/química , Saponinas/farmacocinética , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo
12.
Biochem Biophys Res Commun ; 511(4): 833-839, 2019 04 16.
Artigo em Inglês | MEDLINE | ID: mdl-30846208

RESUMO

ß-N-acetylhexosaminidases from Akkermansia muciniphila was reported to perform the crystal structure with GlcNAc complex, which proved to be the substrate of Am2301. Domain II of Am2301 is consisted of amino acid residues 111-489 and is folded as a (ß/α)8 barrel with the active site combined of the glycosyl hydrolases. Crystallographic evidence showed that Asp-278 and Glu-279 could be the catalytic site and Tyr-373 may plays a role on binding the substrate. Moreover, Am2301 prefers to bind Zn ion, which similar to other GH 20 family. Enzyme activity and kinetic parameters of wild-type Am2301 and mutants proved that Asp-278 and Glu-279 are the catalytic sites and they play a critical role on the catalytic process. Overall, our results demonstrate that Am2301 and its complex with GlcNAC provide obvious structural evidence for substrate-assisted catalysis. Obviously, this expands our understanding on the mode of substrate-assisted reaction for this enzyme family in Akkermansia muciniphila.


Assuntos
Acetilglucosamina/metabolismo , Proteínas de Bactérias/metabolismo , Verrucomicrobia/metabolismo , beta-N-Acetil-Hexosaminidases/metabolismo , Proteínas de Bactérias/química , Domínio Catalítico , Cristalografia por Raios X , Cinética , Modelos Moleculares , Especificidade por Substrato , Verrucomicrobia/química , Verrucomicrobia/enzimologia , beta-N-Acetil-Hexosaminidases/química
13.
Bioresour Technol ; 278: 99-107, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30684729

RESUMO

An efficient way to solve the environmental pollution deriving from hydrolyzed polyacrylamide (HPAM)-containing drilling wastewater is urgent. This work adopted a novel method coupling Fenton oxidation with sequencing batch reactor (SBR) to treat gas-field drilling wastewater successively. This Fenton-SBR process reduced COD, HPAM, NH4+-N and total phosphorus (TP) concentrations of drilling wastewater by 98.35%, 87.58%, 94.50% and 93.52%, respectively. While simulated HPAM wastewater with similar HPAM concentration to Fenton-oxidized drilling wastewater was treated only by biological process, and the COD and HPAM removal efficiencies reached 78.26% and 62.95%. The result indicates that the biodegradability of the drilling wastewater was enhanced after Fenton oxidation. Moreover, the analysis on microbial community structure indicates the dominant bacteria in treated drilling wastewater were different from that in treated simulated-wastewater. It can be considered the Fenton-SBR process possesses potential to be applied to treating the drilling wastewater.


Assuntos
Resinas Acrílicas/metabolismo , Eliminação de Resíduos Líquidos/métodos , Águas Residuárias/química , Biodegradação Ambiental , Peróxido de Hidrogênio/química , Hidrólise , Ferro/química , Campos de Petróleo e Gás , Oxirredução , Eliminação de Resíduos Líquidos/instrumentação
14.
Anal Sci ; 35(4): 441-448, 2019 Apr 10.
Artigo em Inglês | MEDLINE | ID: mdl-30606912

RESUMO

In this paper, an ultrasensitive electrochemical biosensor based on carboxylated multi-walled carbon nanotube/molybdenum disulfide composites (MWCNTs-COOH/MoS2) for the detection of KRAS gene is described. An easy, low-cost method, named one-step hydrothermal, was used for the synthesize of MWCNTs-COOH/MoS2 nanocomposites, and scanning electronic microscopy (SEM), high resolution transmission electron microscopy (HRTEM), Fourier transform infrared spectroscopy (FT-IR), X-ray photoelectron spectroscopy (XPS) and X-ray diffraction (XRD) were used for characterizing the prepared composites. Furthermore, cyclic voltammetry (CV) and differential pulse voltammetry (DPV) were employed for an electrochemical performance study of this biosensor. Under optimal conditions, the detection limit of target DNA achieved down to 3 fM (S/N = 3) with high sensitivity; the linear range with the logarithm of the concentrations of target DNA varied from 1.0 × 10-14 to 1.0 × 10-7 M. Finally, the practicality of our proposed sensor was verified by a determination of the KRAS gene in human serum samples with good accuracy and high precision due to the excellent conductivity and large active surface area of the MWCNTs-COOH/MoS2 nanocomposites. This proposed biosensor thus provides a practical method for the rapid and sensitive analysis of gene detection.


Assuntos
Técnicas Biossensoriais , Ácidos Carboxílicos/química , DNA/química , Dissulfetos/química , Técnicas Eletroquímicas , Molibdênio/química , Nanotubos de Carbono/química , Proteínas Proto-Oncogênicas p21(ras)/genética , Humanos , Tamanho da Partícula , Proteínas Proto-Oncogênicas p21(ras)/sangue , Propriedades de Superfície
15.
Plant Sci ; 272: 131-141, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29807583

RESUMO

Ethylene signaling converges on the ETHYLENE-INSENSITIVE3 (EIN3)/EIN3-like (EIL) transcription factors to regulate a wide range of developmental processes in plants. EBF1/2 (EIN3-binding F-box protein 1 and 2) negatively regulate the ethylene signaling pathway by mediating the degradation of EIN3/EIL proteins. We uncovered previously that SlEBF1 and SlEBF2 are involved in ethylene response, plant senescence, and fruit ripening in tomato. The present study reports on the identification of a novel tomato F-box gene, designated as SlEBF2-like due that its encoded protein is greater similarity to SlEBF2. The SlEBF2-like promoter region contains three ethylene-response elements (EREs). SlEBF2-like is upregulated by ethylene and downregulated by ethylene inhibitors in tomato seedlings. It is dynamically expressed in flowers during bud-to-anthesis and anthesis-to-post-anthesis transitions, and at the onset of fruit ripening, suggesting its role in these situations where ethylene is required for flower opening and fruit ripening. SlEBF2-like overexpression leaded to fruit elongation, caused ripening and color change to start from fruit bottom and expand gradually to the pedicel, and strongly delayed fruit development and ripening in tomato. Our study indicates that the novel EBF gene, SlEBF2-like, is involved in fruit development and ripening via regulating the ethylene response in tomato.


Assuntos
Proteínas F-Box/metabolismo , Frutas/crescimento & desenvolvimento , Lycopersicon esculentum/metabolismo , Proteínas de Plantas/metabolismo , Etilenos/metabolismo , Proteínas F-Box/genética , Proteínas F-Box/fisiologia , Frutas/metabolismo , Genes de Plantas/genética , Genes de Plantas/fisiologia , Lycopersicon esculentum/genética , Lycopersicon esculentum/crescimento & desenvolvimento , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/fisiologia , Plantas Geneticamente Modificadas/genética , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sequência de DNA , Transdução de Sinais/fisiologia
16.
Parasit Vectors ; 10(1): 585, 2017 Nov 23.
Artigo em Inglês | MEDLINE | ID: mdl-29169404

RESUMO

BACKGROUND: Congenital infection of Toxoplasma gondii is an important factor causing birth defects. The neural stem cells (NSCs) are found to be one of the target cells for the parasite during development of the brain. As a key virulence factor of the parasite that hijacks host cellular functions, ROP18 has been demonstrated to mediate the inhibition of host innate and adaptive immune responses through specific binding different host immunity related molecules. However, its pathogenic actions in NSCs remain elusive. RESULTS: In the present study, ROP18 recombinant adenovirus (Ad-ROP18) was constructed and used to infect C17.2 NSCs. After 3d- or 5d-culture in differentiation medium, the differentiation of C17.2 NSCs and the activity of the Wnt/ß-catenin signaling pathway were detected. The results showed that the protein level of ßIII-tubulin, a marker of neurons, in the Ad-ROP18-transfected C17.2 NSCs was significantly decreased, indicating that the differentiation of C17.2 NSCs was inhibited by the ROP18. The ß-catenin level in the Ad-ROP18-transfected C17.2 NSCs was found to be lower than that in the Ad group. Also, neurogenin1 (Ngn1) and neurogenin2 (Ngn2) were downregulated significantly (P < 0.05) in the Ad-ROP18-transfected C17.2 NSCs compared to the Ad group. Accordingly, the TOP flash/FOP flash dual-luciferase report system showed that the transfection of Ad-ROP18 decreased the Wnt/ß-catenin pathway activity in the C17.2 NSCs. CONCLUSIONS: The inhibition effect of the ROP18 from T. gondii (TgROP18) on the neuronal differentiation of C17.2 NSCs was at least partly mediated through inhibiting the activity of the Wnt/ß-catenin signaling pathway, eventually resulting in the downregulation of Ngn1 and Ngn2. The findings help to better understand potential mechanisms of brain pathology induced by TgROP18.


Assuntos
Diferenciação Celular , Células-Tronco Neurais/parasitologia , Proteínas Serina-Treonina Quinases/genética , Toxoplasma/genética , Adenoviridae/genética , Adenoviridae/fisiologia , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Camundongos , Proteínas do Tecido Nervoso/genética , Células-Tronco Neurais/fisiologia , Células-Tronco Neurais/virologia , Toxoplasma/química , Transfecção , Tubulina (Proteína)/análise , Fatores de Virulência , Via de Sinalização Wnt/genética
17.
Int J Nanomedicine ; 12: 4789-4803, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28740385

RESUMO

Although tremendous efforts have been put into the treatment of infectious diseases to prevent epidemics and mortality, it is still one of the major health care issues that have a profound impact on humankind. Therefore, the development of specific, sensitive, accurate, rapid, low-cost, and easy-to-use diagnostic tools is still in urgent demand. Nanodiagnostics, defined as the application of nanotechnology to medical diagnostics, can offer many unique opportunities for more successful and efficient diagnosis and treatment for infectious diseases. In this review, we provide an overview of the nanodiagnostics for infectious diseases from nanoparticle-based, nanodevice-based, and point-of-care test (POCT) platforms. Most importantly, emphasis focused on the recent trends in the nanotechnology-based POCT system. The current state-of-the-art and most promising point-of-care nanodiagnostic technologies, including miniaturized diagnostic magnetic resonance platform, magnetic barcode assay system, cell phone-based polarized light microscopy platform, cell phone-based dongle platform, and paper-based POCT platform, for infectious diseases were fully examined. The limitations, challenges, and future trends of the nanodiagnostics in POCTs for infectious diseases are also discussed.


Assuntos
Doenças Transmissíveis/diagnóstico , Sistemas Automatizados de Assistência Junto ao Leito/tendências , Humanos , Magnetismo , Nanomedicina/métodos , Nanomedicina/tendências , Nanopartículas
18.
Mol Biosyst ; 13(6): 1182-1192, 2017 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-28470264

RESUMO

Rheumatoid arthritis (RA) is an autoimmune disease with an unknown etiology, occurring in approximately 1.0% of general population. More and more studies have suggested that long non-coding RNAs (lncRNAs) could play important roles in various biological processes and be associated with the pathogenesis of different kinds of diseases including RA. Although a large number of lncRNAs have been found, our knowledge of their function and physiological/pathological significance is still in its infancy. In order to reveal functional lncRNAs and identify the key lncRNAs in RA, we reconstructed a global triple network based on the competitive endogenous RNA (ceRNA) theory using the data from National Center for Biotechnology Information Gene Expression Omnibus and our previous paper. Meanwhile, Gene Ontology (GO) and pathway analysis were performed using Cytoscape plug-in BinGO and Database for Annotation, Visualization, and Integration Discovery (DAVID), respectively. We found that the lncRNA-miRNA-mRNA network was composed of 7 lncRNA nodes, 90 mRNA nodes, 24 miRNA nodes, and 301 edges. The functional assay showed that 147 GO terms and 23 pathways were enriched. In addition, three lncRNAs (S5645.1, XR_006437.1, J01878) were highly related to RA, and therefore, were selected as key lncRNAs. This study suggests that specific lncRNAs are associated with the development of RA, and three lncRNAs (S5645.1, XR_006437.1, J01878) could be used as potential diagnostic biomarkers and therapeutic targets.


Assuntos
Artrite Reumatoide/metabolismo , MicroRNAs/metabolismo , RNA Mensageiro/metabolismo , Artrite Reumatoide/genética , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica/genética , Ontologia Genética , Humanos , Análise de Sequência com Séries de Oligonucleotídeos , RNA Longo não Codificante/metabolismo
19.
Oncotarget ; 8(14): 22927-22935, 2017 Apr 04.
Artigo em Inglês | MEDLINE | ID: mdl-28206971

RESUMO

AIMS: To investigate the association of single nucleotide polymorphisms (SNPs) within vascular endothelial growth factor (VEGF) gene polymorphisms, additional gene- gene and gene- smoking interactions with bladder cancer risk. RESULTS: Bladder cancer risk was significantly higher in carriers of the rs699947- A allele within VEGF gene than those with rs699947- CC genotype (CA+ AA versus CC), adjusted OR (95%CI) = 1.70 (1.16-2.31), and higher in carriers of the rs833052- A allele of within VEGF gene than those with rs833052- CC genotype (CA+ AA versus CC), adjusted OR (95%CI) = 1.65 (1.23-2.12). GMDR analysis indicated a potential interaction between rs2010963 and smoking on bladder cancer risk. Current smokers with rs2010963- GC+CC genotype within VEGF gene have the highest bladder cancer risk, compared to never smokers with rs2010963- GG genotype within VEGF gene, OR (95%CI) = 3.25 (1.71-4.83). Haplotype containing the rs2010963- C and rs833052- A alleles were associated with a statistically increased bladder cancer risk, OR (95%CI) = 2.21 (1.12-3.42). MATERIALS AND METHODS: Generalized multifactor dimensionality reduction (GMDR) was used to screen the best interaction combination among SNPs and smoking. Logistic regression was performed to investigate association of 6 SNPs within VEGF gene, additional gene- gene and gene- smoking interaction with bladder cancer risk. CONCLUSIONS: We found that the A allele of rs699947 and the A allele of rs833052 within VEGF gene, interaction between rs2010963 and smoking, haplotype containing the rs2010963- C and rs833052- A alleles were all associated with increased bladder cancer risk.


Assuntos
Fumar/genética , Neoplasias da Bexiga Urinária/etiologia , Fator A de Crescimento do Endotélio Vascular/genética , Alelos , Feminino , Frequência do Gene , Interação Gene-Ambiente , Predisposição Genética para Doença , Genótipo , Haplótipos , Humanos , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único , Fumar/efeitos adversos , Neoplasias da Bexiga Urinária/genética
20.
Bioresour Technol ; 224: 531-535, 2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-27838320

RESUMO

As the key chiral precursor of Crizotinib (S)-1-(2,6-dichloro-3-fluorophenyl) phenethyl alcohol can be prepared from 1-(2,6-dichloro-3-fluorophenyl) acetophenone by the reductive coupling reactions of alcohol dehydrogenase (ADH) and glucose dehydrogenases (GDH). In this work the heterologous expression plasmids harbouring the encoding genes of ADH and GDH were constructed respectively and co-expressed in the same E. coli strain. After optimization, a co-cross-linked enzyme aggregates (co-CLEAs) of both ADH and GDH were prepared from crude enzyme extracts by cross-linking with the mass ratio of Tween 80, glutaraldehyde and total protein (0.6:1:2) which rendered immobilized biocatalysts that retained 81.90% (ADH) and 40.29% (GDH) activity retention. The ADH/GDH co-CLEAs show increased thermal stability and pH stability compared to both enzymes. The ADH/GDH co-CLEAs also show 80% (ADH) and 87% (GDH) residual activity after seven cycles of repeated use. These results make the ADH/GDH co-CLEAs a potential biocatalyst for the industrial preparation of (S)-1-(2,6-dichloro-3-fluorophenyl) phenethyl alcohol.


Assuntos
Álcool Desidrogenase/química , Glucose 1-Desidrogenase/química , Engenharia de Proteínas/métodos , Proteínas Recombinantes/química , Álcool Desidrogenase/genética , Álcool Desidrogenase/metabolismo , Crizotinibe , Reagentes para Ligações Cruzadas/química , Estabilidade Enzimática , Enzimas Imobilizadas/genética , Enzimas Imobilizadas/metabolismo , Escherichia coli/enzimologia , Escherichia coli/genética , Glucose 1-Desidrogenase/genética , Glucose 1-Desidrogenase/metabolismo , Glutaral/química , Álcool Feniletílico/análogos & derivados , Álcool Feniletílico/metabolismo , Polissorbatos/química , Pirazóis/química , Piridinas/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo
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