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1.
Lasers Med Sci ; 2021 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-33392781

RESUMO

We aimed to investigate the mechanism and effect of photodynamic treatment mediated by 5-aminoketovalerate (5-ALA-PDT) on human ovarian cancer cells (OVCAR3 cells) and to provide a theoretical basis for the subsequent experimental step in vivo. Human ovarian cancer OVCAR3 cells were randomly divided into four groups: control group, laser irradiation alone group, photosensitizer alone group, and photodynamic treatment group. Alterations in cell morphology were observed with an inverted light microscope; cell viability was examined by CCK-8 assays. The ROS content and apoptosis rate were examined by flow cytometry analysis. Western blot was used to detect the expression of apoptosis-related proteins, such as caspase-3, Bax, and Bcl-2, and the expression of cleaved caspase-3 in live cells was detected by a cleaved caspase-3 assay kit. Inverted light microscopy showed alterations in cell morphology in different stages. Comparison with the three other groups indicated that tumor cell proliferation was significantly decreased in the photodynamic treatment group (P < 0.05). Flow cytometry analysis revealed that the content of ROS was higher in the photodynamic group than in the other three groups, and the apoptosis rate was higher in the photodynamic treatment group. The difference compared with the other three groups was statistically significant (P < 0.001). The western blot results indicated that the protein expression of Bcl-2 and caspase-3 was decreased in the photodynamic treatment group, and the protein expression level of Bax was increased (P < 0.05). The expression of cleaved caspase-3 was increased in the photodynamic treatment group compared with the other groups according to the data obtained with a microplate reader. Thus, our results demonstrated that the apoptosis and viability of OVCAR3 cells are altered in response to 5-ALA-PDT; however, no remarkable effects were observed in ovarian cancer cells treated with laser irradiation or photosensitizer alone. 5-ALA-PDT can significantly inhibit the growth of human ovarian cancer cells, and the mechanism of this effect is related to the tumor cell apoptosis mediated by the downregulation of Bcl-2 and caspase-3 and upregulation of Bax protein expression.

2.
Biomed Pharmacother ; 134: 111168, 2021 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-33395598

RESUMO

Axonal demyelination is a consistent pathological characteristic of Spinal cord injury (SCI). Promoting differentiation of oligodendrocytes is of importance for remyelination. Conversion of reactive astrocytes with stem cell potential to oligodendrocytes is proposed as an innovative strategy for SCI repair. Neuregulin-1 (Nrg1) plays an essential role in the differentiation of oligodendrocytes. Therefore, it's a potential treatment for demyelination in SCI that using Nrg1 to drive reactive astrocytes toward oligodendrocyte lineage cells. In this study, tumor necrosis factor-α (TNF-α) was used to induce dedifferentiation of primary rat spinal cord astrocytes into reactive astrocytes and Nrg1 was used to induce astrocytes in vitro and in vivo. The results showed that astrocytes treated with TNF-α expressed immaturity markers CD44 and Musashi1 at mRNA and protein levels, indicating that TNF-α induced the stem cell state of astrocytes. Nrg1 induced reactive astrocytes to express oligodendrocyte markers PDGFR-α and O4 at mRNA and protein levels, indicating that Nrg1 directly converts reactive astrocytes toward oligodendrocyte lineage cells. Moreover, upregulation of PI3K-AKT-mTOR signaling activation in response to Nrg1 was observed. In rats with SCI, intrathecal treatment with Nrg1 converted reactive astrocytes to oligodendrocyte lineage cells, inhibited astrogliosis, promoted remyelination, protected axons and eventually improved BBB score. All the biological effects of Nrg1 were significantly reversed by the co-administration of Nrg1 and ErbB inhibitor, suggesting that Nrg1 functioned through the receptor ErbB. Our findings indicate that Nrg1 is sufficient to trans-differentiate reactive astrocytes to oligodendrocytes via the PI3K-AKT-mTOR signaling pathway and repair SCI. Delivery of Nrg1 for the remyelination processes could be a promising strategy for spinal cord repair.

3.
J Nanosci Nanotechnol ; 21(3): 1672-1677, 2021 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-33404432

RESUMO

Microfluidic chips made by traditional materials (glass and silicon) are still important for fluorescence tests, biocompatible experiments, and high temperature applications. However, the majority of the present bonding methods suffer from ultra-clean requirement, complicated fabrication process, and low production efficiency. In the present work, an Electrohydrodynamic printing assist bonding method was proposed. By this method, the ultraviolet-cured-glue dots were printed onto the silicon substrate, and then the patterned glass and silicon substrate can be bonded together at room temperature. The influence of printing condition (nozzle inner-diameter, applied voltage, printing height, and flow rate) on the diameter of printed dot was analyzed by experiments. By the optimized printing condition, the glass-silicon microfluidic chip can be well bonded. The bonding strength and leakage test demonstrated the high bonding quality of the microfluidic chip (bonding strength of 28 MPa and leakage pressure of 3.5 MPa).

4.
Mikrochim Acta ; 188(1): 27, 2021 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-33404824

RESUMO

Caffeine naturally occurs in tea and cocoa, which is also used as an additive in beverages and has pharmacological effects such as refreshing, antidepressant, and digestion promotion, but excessive caffeine can cause harm to the human body. In this work, based on the specific response between nano zinc 5, 10, 15, 20-tetra(4-pyridyl)-21H-23H-porphine (nano ZnTPyP)-CdTe quantum dots (QDs) and caffeine, combined with chemometrics, a visual paper-based sensor was constructed for rapid and on-site detection of caffeine. The fluorescence of QDs can be quenched by nano ZnTPyP. When caffeine is added to the system, it can pull nano ZnTPyP off the surface of the QDs to achieve fluorescence recovery through electrostatic attraction and nitrogen/zinc coordination. The detection range is 5 × 10-11~3 × 10-9 mol L-1, and the detection limit is 1.53 × 10-11 mol L-1 (R2 = 0.9990) (S/N = 3). The paper-based sensor constructed exhibits good results in real samples, such as tea water, cell culture fluid, newborn bovine serum, and human plasma. Therefore, the sensor is expected to be applied to the rapid instrument-free detection of caffeine in food and biological samples.Graphical abstract.

5.
Artigo em Inglês | MEDLINE | ID: mdl-33400483

RESUMO

Cataluminescence is an attractive oxydic luminescence on the gas-solid interface, and metal-oxide@MOF core@shell architectures show great potential for cataluminescence sensing due to their integrated synergistic effect from core and shell components. However, restricting the direct nucleation and growth of metal-organic frameworks (MOFs) on the topologically distinct surface of metal oxides is a great challenge, owing to the high interface energy from the topology mismatch. Herein, for the first time, a novel liquid-phase concentration-controlled nucleation strategy is exploited to induce the direct assembly of a ZIF-8 layer on the surface of CeO2 nanospheres without any sacrificial templates or further surface modifications. The results show that the construction of the CeO2@ZIF-8 core@shell architecture can be accomplished within 1 min under the mediation of boosted nucleation kinetics. Furthermore, the universality of this developed strategy is demonstrated by the encapsulation of other metal-oxide cores such as magnetic Fe3O4 and ZnCo2O4 core particles with a ZIF-8 shell. Notably, compared to the pure CeO2 and ZIF-8, the obtained CeO2@ZIF-8 nanocomposite exhibits enhanced analytical performance for the cataluminescence sensing of propanal, in which the shell acts as the major catalytic reaction center, while the core contributes to further improving the catalytic efficiency. The proposed facile synthesis strategy with excellent simplicity, rapidity, and universality brings new insights into the engineering of core@shell advanced functional materials with mismatched topologies for catering to the diverse application demands.

6.
Nat Commun ; 12(1): 155, 2021 01 08.
Artigo em Inglês | MEDLINE | ID: mdl-33420071

RESUMO

Dual oxidases (DUOXs) produce hydrogen peroxide by transferring electrons from intracellular NADPH to extracellular oxygen. They are involved in many crucial biological processes and human diseases, especially in thyroid diseases. DUOXs are protein complexes co-assembled from the catalytic DUOX subunits and the auxiliary DUOXA subunits and their activities are regulated by intracellular calcium concentrations. Here, we report the cryo-EM structures of human DUOX1-DUOXA1 complex in both high-calcium and low-calcium states. These structures reveal the DUOX1 complex is a symmetric 2:2 hetero-tetramer stabilized by extensive inter-subunit interactions. Substrate NADPH and cofactor FAD are sandwiched between transmembrane domain and the cytosolic dehydrogenase domain of DUOX. In the presence of calcium ions, intracellular EF-hand modules might enhance the catalytic activity of DUOX by stabilizing the dehydrogenase domain in a conformation that allows electron transfer.

8.
Stem Cell Res Ther ; 12(1): 50, 2021 Jan 09.
Artigo em Inglês | MEDLINE | ID: mdl-33422134

RESUMO

BACKGROUND: Muscle stem cells (MuSCs) are absolutely required for the formation, repair, and regeneration of skeletal muscle tissue. Increasing evidence demonstrated that tissue stem cells, especially mesenchymal stem cells (MSCs), can exert therapeutic effects on various degenerative and inflammatory disorders based on their immunoregulatory properties. Human mesenchymal stem cells (hMSCs) treated with interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) were reported to possess anti-inflammatory functions by producing TNF-stimulated gene 6 (TSG-6). However, whether human muscle stem cells (hMuSCs) also possess TSG-6 mediated anti-inflammatory functions has not been explored. METHODS: The ulcerative colitis mouse model was established by subjecting mice to dextran sulfate sodium (DSS) in drinking water for 7 days. hMuSCs were pretreated with IFN-γ and TNF-α for 48 h and were then transplanted intravenously at day 2 of DSS administration. Body weights were monitored daily. Indoleamine 2,3-dioxygenase (IDO) and TSG-6 in hMuSCs were knocked down with short hairpin RNA (shRNA) and small interfering RNA (siRNA), respectively. Colon tissues were collected for length measurement and histopathological examination. The serum level of IL-6 in mice was measured by enzyme-linked immunosorbent assay (ELISA). Real-time PCR and Western blot analysis were performed to evaluate gene expression. RESULTS: hMuSCs treated with inflammatory factors significantly ameliorated inflammatory bowel disease (IBD) symptoms. IDO and TSG-6 were greatly upregulated and required for the beneficial effects of hMuSCs on IBD. Mechanistically, the tryptophan metabolites, kynurenine (KYN) or kynurenic acid (KYNA) produced by IDO, augmented the expression of TSG-6 through activating their common receptor aryl hydrocarbon receptor (AHR). CONCLUSION: Inflammatory cytokines-treated hMuSCs can alleviate DSS-induced colitis through IDO-mediated TSG-6 production.

9.
Phytomedicine ; 81: 153428, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33341025

RESUMO

BACKGROUND: Idiopathic pulmonary fibrosis (IPF) is a fatal and progressive fibrotic lung disease lacking a validated and effective therapy. Aberrant activation of the Wnt/ß-catenin signaling cascade plays the key role in the pathogenesis of IPF. Betulinic acid is a natural pentacyclic triterpenoid molecule that has excellent antitumor and antiviral activities. HYPOTHESIS: We hypothesized that BA has an anti-pulmonary fibrosis effect mediated by the suppression of the Wnt/ß-catenin pathway. Study design Pulmonary fibrosis markers were detected in vitro and in vivo to confirm the antifibrotic effect of BA. The Wnt/ß-catenin pathway-related proteins were overexpressed to determine the effect of BA on Wnt signaling. METHODS AND RESULTS: BA dose-dependently inhibited Wnt3a-induced fibroblast activation in vitro. Moreover, BA decreased Wnt3a- and LiCl-induced transcriptional activity, as assessed by the TOPFlash assay in fibroblasts, and repressed the expression of the Wnt target genes cyclin D1, axin 2, and S100A4. Further investigation indicated that BA restrained the nuclear accumulation of ß-catenin, mainly by increasing the phospho-ß-catenin ratio (S33/S37/T41 and S45), inhibited the phosphorylation of DVL2 and LRP, and decreased the levels of Wnt3a and LRP6. In agreement with the results of the in vitro assays, the in vivo experiments indicated that BA significantly decreased bleomycin-induced pulmonary fibrosis in mice and suppressed myofibroblast activation by inhibiting Wnt/ß-catenin signaling. CONCLUSION: BA may directly interfere with the Wnt/ß-catenin pathway to subsequently repress myofibroblast activation and pulmonary fibrosis.

10.
Brain Behav Immun ; 91: 615-626, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33035633

RESUMO

Lysophosphatidic acid receptor 1 (LPA1) plays a critical role in proinflammatory processes in the central nervous system by modulating microglia activation. The aim of this study was to explore the anti-inflammatory effects and neurological function improvement of LPA1 inhibition after intracerebral haemorrhage (ICH) in mice and to determine whether prostaglandin E2 (PGE2), E-type prostaglandin receptor 2 (EP2), and NADPH oxidase 2 (NOX2) signalling are involved in LPA1-mediated neuroinflammation. ICH was induced in CD1 mice by autologous whole blood injection. AM966, a selective LPA1 antagonist, was administered by oral gavage 1 h and 12 h after ICH. The LPA1 endogenous ligand, LPA was administered to verify the effect of LPA1 activation. To elucidate potential inflammatory mechanisms of LPA1, the selective EP2 activator butaprost was administered by intracerebroventricular injection with either AM966 or LPA1 CRISPR knockout (KO). Water content of the brain, neurobehavior, immunofluorescence staining, and western blot were performed. After ICH, EP2 was expressed in microglia whereas LPA1 was expressed in microglia, neurons, and astrocytes, which peaked after 24 h. AM966 inhibition of LPA1 improved neurologic function, reduced brain oedema, and suppressed perihematomal inflammatory cells after ICH. LPA administration aggravated neurological deficits after ICH. AM966 treatment and LPA1 CRISPR KO both decreased the expressions of PGE2, EP2, NOX2, NF-κB, TNF-α, IL-6, and IL-1ß expressions after ICH, which was reversed by butaprost. This study demonstrated that inhibition of LPA1 attenuated neuroinflammation caused by ICH via PGE2/EP2/NOX2 signalling pathway in mice, which consequently improved neurobehavioral functions and alleviated brain oedema. LPA1 may be a promising therapeutic target to attenuate ICH-induced secondary brain injury.

11.
Enzyme Microb Technol ; 143: 109720, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33375980

RESUMO

Operational stability under high temperature is required for enzyme application in industrial processes. Error-prone PCR and B-factor analysis were employed to enhance the thermostability of a xylanase from GH family 11 in this study. Based on the top 10 mutants screened from the random mutation libraries, mutant Xyn371 was derived from the optimal mutant Xyn370 by integrating the beneficial residues identified in the other 9 screened mutants. Subsequently, a best-saturation mutant Xyn372 originated from Xyn371 was selected with a 60-min half-life at 70 °C (0.5-min half-life for the wild-type enzyme). According to the site-saturated mutagenesis of 10 residues with higher B-factors in Xyn372, mutants Xyn375 and Xyn376 were screened; their half-lives at 70 °C were 410 and 360 min, respectively. The substituted residues located in the "palm" region of the N-terminus and the newly generated hydrogen bonds in the mutants might contribute to improved thermostability. The significantly improved thermostability of mutants will pave the way for applications in different industrial areas.

12.
Interact Cardiovasc Thorac Surg ; 32(1): 156-158, 2021 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-33221884

RESUMO

Blunt chest trauma can cause a variety of cardiac injuries, either immediately or days after the trauma. We report a case of traumatic ventricular septal defect and ribbonlike left ventricular aneurysm, which was diagnosed 15 years after the initial blunt chest trauma. It was successfully repaired using the endoventricular patch technique with a satisfactory 1-year follow-up result.

13.
Exp Cell Res ; 398(2): 112389, 2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-33221316

RESUMO

Ischemia-reperfusion (I/R) injury is a multifactorial process triggered when an organ is subjected to transiently reduced blood supply. The result is a cascade of pathological complications and organ damage due to the production of reactive oxygen species following reperfusion. The present study aims to evaluate the role of activated calcium-sensing receptor (CaR)-cystathionine γ-lyase (CSE)/hydrogen sulfide (H2S) pathway in I/R injury. Firstly, an I/R rat model with CSE knockout was constructed. Transthoracic echocardiography, TTC and HE staining were performed to determine the cardiac function of rats following I/R Injury, followed by TUNEL staining observation on apoptosis. Besides, with the attempt to better elucidate how CaR-CSE/H2S affects I/R, in-vitro culture of human coronary artery endothelial cells (HCAECs) was conducted with gadolinium chloride (GdCl3, a CaR agonist), H2O2, siRNA against CSE (siCSE), or W7 (a CaM inhibitor). The interaction between CSE and CaM was subsequently detected. Plasma oxidative stress indexes, H2S and CSE, and apoptosis-related proteins were all analyzed following cell apoptosis. We found that H2S elevation led to the improvement whereas CSE knockdown decreased cardiac function in rats with I/R injury. Moreover, oxidative stress injury in I/R rats with CSE knockout was aggravated, while the increased expression of H2S and CSE in the aortic tissues resulted in alleviated the oxidative stress injury. Moreover, increased H2S and CSE levels were found to inhibit cell apoptotic ability in the aortic tissues after I/R injury, thus attenuating oxidative stress injury, accompanied by inhibited expression of apoptosis-related proteins. In HCAECs following oxidative stress treatment, siCSE and CaM inhibitor were observed to reverse the protection of CaR agonist. Coimmunoprecipitation assay revealed the interaction between CSE and CaM. Taken together, all above-mentioned data provides evidence that activation of the CaR-CSE/H2S pathway may confer a potent protective effect in cardiac I/R injury.

14.
Int J Biol Macromol ; 166: 1-8, 2021 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-33220380

RESUMO

The carboxymethylated (1 â†’ 6)-α-dextran (CM-dex) was synthesized by introducing carboxymethyl groups at different degrees of substitution (DS). The resulting dex1-1, dex2-1, dex3-1, and dex4-1 products had degrees of substitution of 0.57, 0.78, 1.13, and 1.25, respectively. The dex3-1 showed the highest glass transition temperature (Tg) of 215.96 °C, whereas Tg of pure dextran was 149.83 °C. TGA results indicated that the residual loss was reduced along with the increase of DS in the high-temperature region (450-600 °C). Besides, the CM-dex had stronger scavenging capacity against OH radicals but lower scavenging capacity for DPPH (1,1-diphenyl-2-picrylhydrazyl) radicals compared to that of pure dextran. The carboxymethylation of (1 â†’ 6)-α-dextran will extend the applications for modified dextran.

15.
Biosens Bioelectron ; 174: 112828, 2021 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-33250335

RESUMO

Wearable electrochemical sensors have attracted tremendous attention in recent years. Significant progress has been achieved, particularly in device integration. Most wearable devices are integrated on thin-film polymer, however, less attention is paid to the sweat flow at human-device interfaces, which is of great significance for continuous real-time analysis and long-term skin comfort. Here, we reported a low-cost, freestanding and disposable highly integrated sensing paper (HIS paper) for real-time analysis of sweat. By using a simple printing process, the HIS paper combining hydrophobic protecting wax, conducting electrodes, and the incorporated MXene/methylene blue (Ti3C2Tx/MB) active materials was assembled. In particular, the printed paper was folded into a multi-layer structure, in which a reasonable designed three-dimensional (3D) sweat diffusion path is established by connecting the hydrophilic regions of each layer, providing efficient pathways for the collection and diffusion of sweat along the vertical direction of the folded HIS paper. More importantly, the independent 3D position of three-electrode facilitates the decoration and fixation of enzymes, as well as the accessibility of electrolytes. In addition, a dual-channel electrochemical sensor that can simultaneously detect glucose and lactate with sensitivity of 2.4 nA µM-1 and 0.49 µA mM-1 respectively was produced based on the HIS paper. This HIS paper provides a miniaturized, low-cost and flexible solution for a range of biochemical platforms, including wearable bioelectronics.

16.
Food Chem ; 341(Pt 1): 128218, 2021 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-33035857

RESUMO

In this study, two polysaccharide fractions were isolated from wheat bran by sequential extraction with water and alkaline solution, DEAE Cellulose-52 chromatography and Sephacryl S-400 gel permeation chromatography, they were named as WXA-1 and AXA-1, respectively. Structural analyses indicated that both polysaccharide fractions were heteropolysaccharides, their average molecular weights were 193 kDa and 107 kDa, respectively. The backbone of WXA-1 was â†’ 4)-ß-d-Xylp-(1→, which was substituted at O-3 positions by arabinose, glucose and galactose residues, while the backbone of AXA-1 was â†’ 4)-ß-d-Xylp-(1→, which was mainly substituted at O-3 positions by arabinose. AXA-1 exerted a stronger inhibitory effect on the activities of α-amylase and α-glucosidase compared with WXA-1. Moreover, AXA-1 exhibited a competitive inhibition of α-amylase and a mixed-type noncompetitive inhibition of α-glucosidase. These results suggest that AXA-1 can be used as α-amylase and α-glucosidase inhibitors.


Assuntos
Fibras na Dieta/análise , Inibidores de Glicosídeo Hidrolases/química , Inibidores de Glicosídeo Hidrolases/farmacologia , Polissacarídeos/química , Polissacarídeos/farmacologia , alfa-Amilases/metabolismo , alfa-Glucosidases/metabolismo , Peso Molecular
20.
Am J Respir Cell Mol Biol ; 64(1): 115-125, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33074715

RESUMO

Augmented glycolysis due to metabolic reprogramming in lung myofibroblasts is critical to their profibrotic phenotype. The primary glycolysis byproduct, lactate, is also secreted into the extracellular milieu, together with which myofibroblasts and macrophages form a spatially restricted site usually described as fibrotic niche. Therefore, we hypothesized that myofibroblast glycolysis might have a non-cell autonomous effect through lactate regulating the pathogenic phenotype of alveolar macrophages. Here, we demonstrated that there was a markedly increased lactate in the conditioned media of TGF-ß1 (transforming growth factor-ß1)-induced lung myofibroblasts and in the BAL fluids (BALFs) from mice with TGF-ß1- or bleomycin-induced lung fibrosis. Importantly, the media and BALFs promoted profibrotic mediator expression in macrophages. Mechanistically, lactate induced histone lactylation in the promoters of the profibrotic genes in macrophages, consistent with the upregulation of this epigenetic modification in these cells in the fibrotic lungs. The lactate inductions of the histone lactylation and profibrotic gene expression were mediated by p300, as evidenced by their diminished concentrations in p300-knockdown macrophages. Collectively, our study establishes that in addition to protein, lipid, and nucleic acid molecules, a metabolite can also mediate intercellular regulations in the setting of lung fibrosis. Our findings shed new light on the mechanism underlying the key contribution of myofibroblast glycolysis to the pathogenesis of lung fibrosis.

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