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The inter-satellite laser ranging interferometer is one of the core components of future gravity missions to achieve high ranging precision. This work builds a preliminary breadboard of the off-axis optical bench design, which integrates the merits of the off-axis optical bench design of GRACE Follow-On mission and other on-axis designs. The study finds that the displacement noise between two optical benches has been reduced to 20nm/Hz at a frequency of 10 mHz, and the differential wavefront sensing noise has been suppressed to 10-5rad/Hz at 1 kHz as well. In addition, the tilt-to-length coupling noise related to the piston effect is restricted within 30 µm/rad, and the relative parallelism error of the transmitting beam and receiving beam is less than 4.5%. The results show that this off-axis optical bench design is an important candidate for China's future gravity missions.
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Exposure to mustard gas can cause damage or death to human beings, depending on the concentration and duration. Thus, developing high-performance mustard-gas sensors is highly needed for early warning. Herein, ultrathin WO3 nanosheet-supported Pd nanoparticles hybrids (WO3 NSs/Pd) are prepared as chemiresistive sulfur mustard simulant (e.g., 2-chloroethyl ethyl sulfide, 2-CEES) gas sensors. As a result, the optimal WO3 NSs/Pd-2 (2 wt % of Pd)-based sensor exhibits a high response of 8.5 and a rapid response/recovery time of 9/92 s toward 700 ppb 2-CEES at 260 °C. The detection limit could be as low as 15 ppb with a response of 1.4. Moreover, WO3 NSs/Pd-2 shows good repeatability, 30-day operating stability, and good selectivity. In WO3 NSs/Pd-2, ultrathin WO3 NSs are rich in oxygen vacancies, offer more sites to adsorb oxygen species, and make their size close to or even within the thickness of the so-called electron depletion layer, thus inducing a large resistance change (response). Moreover, strong metal-support interactions (SMSIs) between WO3 NSs and Pd nanoparticles enhance the catalytic redox reaction performance, thereby achieving a superior sensing performance toward 2-CEES. These findings in this work provide a new approach to optimize the sensing performance of a chemiresistive sensor by constructing SMSIs in ultrathin metal oxides.
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Gás de Mostarda , Óxidos , Paládio , Tungstênio , Tungstênio/química , Paládio/química , Gás de Mostarda/análise , Gás de Mostarda/química , Gás de Mostarda/análogos & derivados , Óxidos/química , Limite de Detecção , Nanopartículas Metálicas/química , Nanoestruturas/química , Substâncias para a Guerra Química/análise , Substâncias para a Guerra Química/química , Técnicas Eletroquímicas/métodosRESUMO
The utilization of steel slag for CO2 sequestration is an effective way to reduce carbon emissions. The reactivity of steel slag in CO2 sequestration depends mainly on material and process parameters. However, there are many puzzles in regard to practical applications due to the different evaluations of process parameters and the lack of investigation of material parameters. In this study, 318 samples were collected to investigate the interactive influence of 12 factors on the carbonation reactivity of steel slag by machine learning with SHapley Additive exPlanations (SHAP). Multilayer perceptron (MLP), random forest, and support vector regression models were built to predict the slurry-phase CO2 sequestration of steel slag. The MLP model performed well in terms of prediction ability and generalization with comprehensive interpretability. The SHAP results showed that the impact of the process parameters was greater than that of the material parameters. Interestingly, the iron ore phase of steel slag was revealed to have a positive effect on steel slag carbonation by SHAP analysis. Combined with previous literature, the carbonation mechanism of steel slag was proposed. Quantitative analysis based on SHAP indicated that steel slag had good carbonation reactivity when the mass fractions of "CaO + MgO", "SiO2 + Al2O3", "Fe2O3", and "MnO" varied from 50-55%, 10-15%, 30-35%, and <5%, respectively.
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Dióxido de Carbono , Resíduos Industriais , Resíduos Industriais/análise , Dióxido de Carbono/análise , Aço , Dióxido de Silício , Carbonatos , Aprendizado de MáquinaRESUMO
The inter-satellite laser ranging heterodyne interferometer is vital for future gravity missions to achieve high ranging accuracy. This paper proposes a novel off-axis optical bench design which integrates merits of the off-axis optical bench design of GRACE Follow-On mission and other on-axis designs. This design makes use of lens systems subtly to restrict the tilt-to-length coupling noise and takes advantage of the DWS feedback loop to maintain the transmitting beam and receiving beam anti-parallel. The critical parameters of the optical components are determined and the carrier-to-noise ratio for a single channel of the photoreceiver is calculated to be more than 100 dB-Hz for the high case. The off-axis optical bench design is a potential candidate for China's future gravity missions.
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We generate a macro-pulsed chaotic laser based on pulse-modulated laser diode subject to free space optical feedback, and demonstrate the performance of suppressing backscattering interference and jamming in turbid water. The macro-pulsed chaotic laser with a wavelength of 520â nm as a transmitter is used with a correlation-based lidar receiver to perform an underwater ranging. At the same power consumption, macro-pulsed lasers have higher peak power than in the continuous-wave form, enabling the former to detect longer ranging. The experimental results show that a chaotic macro-pulsed laser has excellent performance of suppressing the backscattering of water column and anti-noise interference compared with traditional pulse laser, especially by multiple accumulations about 10â¼30 times, and target position can still be determined when SNR is -20â dB.
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Silicon-based field effect transistor (FET) sensors with high sensitivity are emerging as powerful sensors for detecting chemical/biological species. Strain engineering has been demonstrated as an effective means to improve the performance of Si-based devices. However, the strain effect on the field-effect sensing property of silicon materials has not been studied yet. Here, we investigate the strain effect on the field-effect sensing property of silicon wires by taking humidity sensing as an example. The humidity sensitivity of FET sensors based on silicon wires increases with increasing tensile strain but decreases with increasing compressive strain. The sensitivity is very responsive to strain with an enhancement factor of 67 for tensile strain. Theoretical analysis shows that the sensitivity variation under different strains is mainly attributed to the change in adsorption energy between silicon wires and water molecules. This work indicates that strain engineering can be an effective route to modulate the field-effect sensing property of Si wires for constructing highly sensitive Si-based FET sensors.
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Chemical warfare agents (CWAs) can be absorbed in polymeric coatings through absorption and permeation, thus presenting a lethal touch and vapor hazards to people. Developing a highly impermeable polymer coating against CWAs, especially against organophosphate CWAs (OPs), is challenging and desirable. Herein, fluorinated epoxy (F-EP) and epoxy (EP) coatings with different cross-link densities were prepared to resist OPs. The effects of the polymer coating structure, including cross-link density, chemical composition and free volume, on the chemical resistance to dimethyl methylphosphonate (DMMP, Soman simulant) were investigated in detail. Meanwhile, the chemical resistance to Soman and VX was examined. The results reveal that the cross-link density is a critical factor in determining the chemical resistance of the coatings. Highly cross-linked EP and F-EP coatings with dense and solid cross-linked networks can fully bar DMMP and OPs permeation during the test time. At low or medium cross-link densities, the EP coating with a lower retention of DMMP exhibited a higher resistance than the F-EP coating due to the lower interaction with DMMP and smaller free-volume holes and lower relative fractional free volume. These results suggest that increasing the cross-link density is a reasonable approach to control the chemical resistance of polymer networks against OPs.
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Due to deadly toxicity and high environmental stability of the nerve agent VX, an efficient decontamination approach is desperately needed in tackling its severe threat to human security. The enzymatic destruction of nerve agents has been generally considered as one of the most effective ways, and here the hydrolysis of VX by phosphotriesterase (PTE) was investigated by extensive QM/MM and MM MD simulations. The hydrolytic cleavage of P-S by PTE is a two-step process with the free energy spans of 15.8 and 26.0 kcal mol-1 for the RP- and SP-enantiomer VX, respectively, and such remarkable stereospecificity of VX enantiomers in the enzymatic degradation is attributed to their conformational compatibility with the active pocket. The structurally less adaptive SP-enantiomer allows one additional water molecule to enter the binuclear zinc center and remarkably facilitates the release of the degraded product. Overall, the rate-limiting steps in the enzymatic degradation of VX by PTE involve the degraded product release of the RP-enantiomer and the enzymatic P-S cleavage of the SP-enantiomer. Further computational analysis on the mutation of selected residues also revealed that H257Y, H257D, H254Q-H257F, and L7ep-3a variants allow more water molecules to enter the active site, which improves the catalytic efficiency of PTE, as observed experimentally. The present work provides mechanistic insights into the stereoselective hydrolysis of VX by PTE and the activity manipulation through the active-site accessibility of water molecules, which can be used for the enzyme engineering to defeat chemical warfare agents.
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Substâncias para a Guerra Química , Agentes Neurotóxicos , Hidrolases de Triester Fosfórico , Domínio Catalítico , Substâncias para a Guerra Química/química , Substâncias para a Guerra Química/metabolismo , Substâncias para a Guerra Química/toxicidade , Descontaminação , Humanos , Hidrólise , Compostos Organotiofosforados , Hidrolases de Triester Fosfórico/química , Hidrolases de Triester Fosfórico/genética , Hidrolases de Triester Fosfórico/metabolismo , ÁguaRESUMO
Two-photon polymerization of a three-dimensional (3D) hydrogel structure has been widely applied in biological tissue engineering. For improving the biocompatibility of hydrogel structures, a new kind of ionic carbazole water-soluble photoinitiator was prepared to realize the fabrication of a 3D hydrogel structure in aqueous phase. 3,6-Bis[2-(1-methyl-pyridinium)vinyl]-9-methyl-carbazole diiodide (BMVMC) and cucurbit[7]uril (CB7) have been employed to generate a complex with better water solubility by host-guest interactions. The binding ratio of the complex was demonstrated to be 1:1 through the characterization of isothermal titration calorimetry (ITC). The two-photon absorption (TPA) cross section of the complex increases to 2500 GM compared with the 750 GM of the BMVMC molecule. Then, an aqueous-phase photoresist was obtained using the CB7/BMVMC complex as the photoinitiator and poly(ethylene glycol) diacrylate (PEGda) as the hydrogel monomer. Two-photon fabrication capability in aqueous phase has been studied using the as-prepared photoresist. A low laser threshold of 3.7 mW as well as a high resolution of 180 nm are achieved. Benefiting from the fluorescence properties of the photoinitiator, we can achieve the confocal fluorescence images without any assistance of fluorescent probes. Subsequently, a 3D engineered hydrogel scaffold microstructure was fabricated by the two-photon polymerization technology, whose biocompatibility was demonstrated by culturing the structure with living cells of L929. The BMVMC-CB7 complex and the as-prepared photoresist are demonstrated to have good biocompatibility, which is prospective for further application in tissue engineering.
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Carbazóis/química , Hidrogéis/química , Alicerces Teciduais/química , Animais , Hidrocarbonetos Aromáticos com Pontes/síntese química , Hidrocarbonetos Aromáticos com Pontes/química , Hidrocarbonetos Aromáticos com Pontes/toxicidade , Carbazóis/síntese química , Carbazóis/efeitos da radiação , Carbazóis/toxicidade , Linhagem Celular , Módulo de Elasticidade , Hidrogéis/síntese química , Hidrogéis/efeitos da radiação , Hidrogéis/toxicidade , Imidazóis/síntese química , Imidazóis/química , Imidazóis/toxicidade , Camundongos , Fótons , Polimerização/efeitos da radiação , Solubilidade , Engenharia Tecidual/métodos , Água/químicaRESUMO
With the increasing application of enzymes in various research fields, the choices of co-solvents in enzymatic preparations which directly related to the catalytic activity have been attracted attention. Thus, researching on the stabilization or destabilization behaviors of enzymes in different solvents is extremely essential. In this study, the structural changes of DhaA in two typical aprotic co-solvents (acetonitrile and tetrahydrofuran) were firstly investigated by molecular dynamics (MD) simulation. The simulation results revealed the strong van der Waals force between co-solvents and DhaA which could induce the structural change of enzyme. Interestingly, the differences of molecular size and the electrostatic force with enzyme of two co-solvents led to quite different influences on DhaA. As for acetonitrile, solvent molecules could penetrate into the catalytic site of DhaA which promoted by the electrostatic interaction. On the contrary, tetrahydrofuran molecules were mainly distributed around the catalytic site due to the relative weak electrostatic interaction and steric resistance effect. It can be concluded that different co-solvent can affect the key domains, substrate pathway and catalytic pocket of DhaA.Communicated by Ramaswamy H. Sarma.
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Hidrolases , Simulação de Dinâmica Molecular , Catálise , Domínio Catalítico , SolventesRESUMO
The efficient immobilization of haloalkane dehalogenase (DhaA) on carriers with retaining of its catalytic activity is essential for its application in environmental remediation. In this work, adsorption orientation and conformation of DhaA on different functional surfaces were investigated by computer simulations; meanwhile, the mechanism of varying the catalytic activity was also probed. The corresponding experiments were then carried out to verify the simulation results. (The simulations of DhaA on SAMs provided parallel insights into DhaA adsorption in carriers. Then, the theory-guided experiments were carried out to screen the best surface functional groups for DhaA immobilization.) The electrostatic interaction was considered as the main impact factor for the regulation of enzyme orientation, conformation, and enzyme bioactivity during DhaA adsorption. The synergy of overall conformation, enzyme substrate tunnel structural parameters, and distance between catalytic active sites and surfaces codetermined the catalytic activity of DhaA. Specifically, it was found that the positively charged surface with suitable surface charge density was helpful for the adsorption of DhaA and retaining its conformation and catalytic activity and was favorable for higher enzymatic catalysis efficiency in haloalkane decomposition and environmental remediation. The neutral, negatively charged surfaces and positively charged surfaces with high surface charge density always caused relatively larger DhaA conformation change and decreased catalytic activity. This study develops a strategy using a combination of simulation and experiment, which can be essential for guiding the rational design of the functionalization of carriers for enzyme adsorption, and provides a practical tool to rationally screen functional groups for the optimization of adsorbed enzyme functions on carriers. More importantly, the strategy is general and can be applied to control behaviors of different enzymes on functional carrier materials.
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Hidrolases/química , Rhodococcus/enzimologia , Adsorção , Enzimas Imobilizadas/química , Enzimas Imobilizadas/metabolismo , Hidrolases/metabolismo , Modelos Moleculares , Eletricidade Estática , Propriedades de SuperfícieRESUMO
Here, we introduced a novel thiourea-based rhodamine compound as a chromo-fluorogenic indicator of nerve agent Soman and its simulant diethyl chlorophosphate (DCP). The synthesized probe N-(rhodamine B)-lactam-2-(4-cyanophenyl) thiourea (RB-CT), which has a rhodamine core linked by a cyanophenyl thiosemicarbazide group, enabled a rapidly and highly sensitive response to DCP with clear fluorescence and color changes. The detection limit was as low as 2 × 10-6 M. The sensing mechanism showed that opening of the spirolactam ring following the phosphorylation of thiosemicarbazides group formed a seven-membered heterocycle adduct, according to MS analysis and TD-DFT calculations. RB-CT exhibited high detecting selectivity for DCP, among other organophosphorus compounds. Moreover, two test kits were employed and successfully used to detect real nerve agent Soman in liquid and gas phase.
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Corantes Fluorescentes/síntese química , Compostos Organofosforados/análise , Rodaminas/química , Soman/análise , Tioureia/química , Substâncias para a Guerra Química/análise , Substâncias para a Guerra Química/química , Corantes Fluorescentes/química , Limite de Detecção , Estrutura Molecular , Agentes Neurotóxicos/análise , Agentes Neurotóxicos/química , Compostos Organofosforados/química , Soman/químicaRESUMO
We have introduced a novel water-soluble two-photon photoinitiator based on the host-guest interaction between 3,6-bis[2-(1-methyl-pyridinium)vinyl]-9-pentyl-carbazole diiodide (BMVPC) and cucurbit[7]uril (CB7) because most of the commercial photoinitiators have poor two-photon initiating efficiency in aqueous solutions. The binding ratio of BMVPC and CB7 was determined as 1:1 by isothermal titration calorimetry and quantum chemical calculation. The formation of the host-guest complex increases the two-photon absorption cross-section about five times, and improves the water solubility required as the photoinitiator for hydrogel fabrication. The BMVPC-CB7 inclusion complex was used as the one-component photoinitiator, and the polyethylene glycol diacrylate with promising biocompatibility was used as the hydrogel monomer to form the aqueous-phase photoresist system applied to two-photon polymerization microfabrication. A relatively low laser threshold of 4.5 mW, a high fabricating resolution of 180 nm, and the true three-dimensional (3D) fabricating capability in the aqueous solution have been obtained by using the as-prepared photoresist system. Finally, 3D engineering hydrogel scaffold microstructures with low toxicity and good biocompatibility have been fabricated and cocultured with living HeLa cells, which demonstrates the potential for further application in tissue engineering.
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Materiais Biocompatíveis , Hidrocarbonetos Aromáticos com Pontes/química , Carbazóis/química , Hidrogéis , Imidazóis/química , Lasers , Alicerces Teciduais/química , Materiais Biocompatíveis/química , Células HeLa , Humanos , Hidrogéis/síntese química , Hidrogéis/química , Microtecnologia , Polietilenoglicóis/química , Engenharia TecidualRESUMO
In this study, two kinds of novel carbazole-based ethynylpyridine salts: 3,6-bis[2-(1-methylpyridinium)ethynyl]-9-pentyl-carbazole diiodide (BMEPC) and 3,6-bis[2-(1-methylpyridinium)ethynyl]-9-methyl-carbazole diiodide (BMEMC) have been employed as photosensitizers owing to their excellent antibacterial activity. These molecules possess symmetric A-π-D-π-A-type structures, which would bring in the unique optical properties. The inhibition zone measurement of a gradient concentration from 0 to 100 µM showed BMEPC and BMEMC photoinduced antibacterial activity against Escherichia coli. Diameters of zone of inhibition were up to 15 and 14 mm under laser irradiations. Under the exposure of the laser of 442 nm with a power density of 20 mW/cm2, the minimum inhibitory concentrations (MICs) of BMEPC on E. coli were between 3.5 and 6.9 µM and that of BMEMC were between 9.4 and 18.8 µM, respectively. In the dark experiments as a control, the MIC value is between 6.9 and 13.8 µM for BMEPC, whereas it is between 187.5 and 225.0 µM for BMEMC. By the comparison of the MIC values of BMEPC and BMEMC with laser irradiation and in dark, the laser-induced toxicity on bacteria is more evident, though both of the derivatives have dark toxicity. With the laser irradiation duration of 30 s and 10 min for BMEPC and BMEMC, respectively, the survival rate of E. coli approximates zero. An antibacterial mechanism has been proposed based on the electron paramagnetic resonance characterization, which indicates that a nitride radical is generated under laser irradiation. The carbazole-based ethynylpyridine photosensitizers would provide high potential for further applications in photodynamic therapy.
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BACKGROUND: Hypertrophic scars and keloids, characterized by over-proliferation of fibroblasts and aberrant formation of the extracellular matrix (ECM), are considered fibrotic diseases. Accumulating evidence indicates that mesenchymal stem cells (MSCs) promote scar-free wound healing and inhibit fibrotic tissue formation, making them a potentially effective therapeutic treatment for hypertrophic scars and keloids. OBJECTIVE: To investigate the paracrine effects of bone marrow derived MSCs (BMSCs) on the biological behavior of hypertrophic scar fibroblasts (HSFs) and keloid fibroblasts (KFs). METHODS: Proliferative and profibrotic phenotype changes of the fibroblasts were analyzed by immunofluorescence staining, in-cell western blot, and real-time PCR. RESULTS: BMSC-conditioned medium inhibited HSF and KF proliferation and migration, but did not induce apoptosis. Interestingly, normal skin fibroblast-conditioned medium exhibited no inhibitory effects on HSF or KF proliferation and migration. Furthermore, BMSC-conditioned medium significantly decreased expression of profibrotic genes, including connective tissue growth factor, plasminogen activator inhibitor-1, transforming growth factor-ß1, and transforming growth factor-ß2, in HSFs and KFs at both transcriptional and translational levels. In contrast, the expression of antifibrotic genes, such as transforming growth factor-ß3 and decorin, was substantially enhanced under the same culture conditions. Finally, we observed that BMSC-conditioned medium suppressed the ECM synthesis in HSFs and KFs, as indicated by decreased expression of collagen I and fibronectin and low levels of hydroxyproline in cell culture supernatant. CONCLUSION: These findings suggest that BMSCs attenuate the proliferative and profibrotic phenotype associated with HSFs and KFs and inhibit ECM synthesis through a paracrine signaling mechanism.
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Cicatriz Hipertrófica/terapia , Matriz Extracelular/metabolismo , Fibroblastos/fisiologia , Queloide/terapia , Células-Tronco Mesenquimais/fisiologia , Comunicação Parácrina , Cicatrização/fisiologia , Adolescente , Adulto , Células da Medula Óssea/citologia , Proliferação de Células , Células Cultivadas , Cicatriz Hipertrófica/metabolismo , Cicatriz Hipertrófica/patologia , Colágeno Tipo I/metabolismo , Fator de Crescimento do Tecido Conjuntivo/antagonistas & inibidores , Meios de Cultivo Condicionados/farmacologia , Decorina/antagonistas & inibidores , Feminino , Fibroblastos/patologia , Fibronectinas/antagonistas & inibidores , Humanos , Queloide/metabolismo , Queloide/patologia , Masculino , Células-Tronco Mesenquimais/efeitos dos fármacos , Pessoa de Meia-Idade , Inibidor 1 de Ativador de Plasminogênio/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Pele/citologia , Pele/metabolismo , Fator de Crescimento Transformador beta1/metabolismo , Fator de Crescimento Transformador beta2/metabolismo , Fator de Crescimento Transformador beta3/metabolismo , Adulto JovemRESUMO
MicroRNAs (miRNAs) are post-transcriptional modulators of gene expression and play an important role in reprogramming process; however, relatively little is known about the underlying regulatory mechanism of miRNAs on how they epigenetically modulate reprogramming and pluripotency. Here, we report that the expression level of microRNA-134 (miR-134) was low in mouse embryonic stem cells (mESCs) but significantly up-regulated during neural differentiation, while down-regulated during the induction of induced pluripotent stem cells (iPSCs) from neural progenitor cells (NPCs). Inhibition of miR-134 by miR-134 sponge promoted the efficiency of reprogramming which also was highly similar to mESCs. On the contrary, up-regulation of miR-134 repressed iPSCs induction. We also found that inhibition of miR-134 promoted the maturation of pre-iPSCs and increased its pluripotency. We also showed that miR-134 can directly target to the pluripotency related factor Methyl-CpG-binding domain protein 3 (Mdb3) 3' untranslated regions (3' UTR) to down-regulate its expression. And Mbd3 was found to promote the induction of iPSCs and could block the repression of reprogramming caused by overexpression of miR-134. This work revealed the critical function of miR-134-Mbd3 axis on regulating reprogramming and pluripotency of iPSCs derived from the NPCs, and might provide an insight into the miR-134-Mbd3 axis on regulating the iPSCs quality for further clinical treatment.
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Proteínas de Ligação a DNA/metabolismo , Células-Tronco Pluripotentes Induzidas/metabolismo , MicroRNAs/metabolismo , Fatores de Transcrição/metabolismo , Regiões 3' não Traduzidas/genética , Animais , Sequência de Bases , Regulação para Baixo/genética , Células-Tronco Pluripotentes Induzidas/citologia , Camundongos , Células-Tronco Neurais/citologia , Células-Tronco Neurais/metabolismoRESUMO
Microenvironment in biology is diverse and complex which has been a great challenge for in vivo imaging materials, and so materials with environmental tolerance and photostability need to be explored. For aggregation-induced emission (AIE) molecules, the fluorescence is closely related to the restricted structure which is directly affected by the microenvironment. Inorganic silica nanoparticles can provide a rigid microenvironment which can stabilize AIE molecules to obtain fluorescent materials with environmental tolerance. Here, stable fluorescent SiO2 nanoparticles (CWQ-11@SiO2 NPs) have been prepared by doping with typical AIE molecules named CWQ-11. CWQ-11@SiO2 NPs have narrow size distribution and spherical morphology with a size of around 50 nm. The fluorescence intensity of CWQ-11@SiO2 NPs is nearly 45.4 times higher than that of free CWQ-11. CWQ-11@SiO2 NPs maintain excellent fluorescence stabilities under various conditions, such as in solutions with different pH values, different viscosities, or continuous irradiation, and even in simulated gastric fluid (SGF). Cellular imaging research represents efficient imaging ability of CWQ-11@SiO2 NPs in two different tumor cells including MCF-7S and HepG-2. All these results demonstrate that the CWQ-11@SiO2 NPs have been successfully prepared and remain stable under different harsh conditions, and have promising potential in imaging, tracing for drugs or diagnosis in complicated biological systems.
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Here, we have introduced a novel biscarbazolylmethane-based cyanine as a two-photon excited fluorescent probe, 6,6'-bis[2-(1-methylpyridinium)vinyl]-bis(9-methyl-carbazol-3-yl)methane diiodide, which has two vinylpyridinium carbazole moieties connected by a non-rigid methylene bridge. This molecule possesses a larger Stokes shift and enhanced two-photon absorption cross-section than the previously reported vinylpyridinium carbazole monocyanine, which is mainly attributed to the "through-space" type intramolecular charge transfer. The low fluorescence quantum yield and 30-fold fluorescence enhancement once binding with calf thymus DNA highlight this molecule as a promising fluorescence light-up probe for DNA. The obvious induced circular dichroism signals have proved that the molecule with soft-connected bis-cationic centers can specifically interact with various DNA structures. Cell viability study shows that the probe has very low cytotoxicity. The probe exhibits high staining selectivity for mitochondria in living HeLa cells. Its capability to stain nuclear DNA has been confirmed by fixed cell staining. Furthermore, the application for two-photon excited fluorescence imaging demonstrates high potential of the probe for nonlinear bioimaging with 3D resolution.
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AIM: To study the effect of retinoid X receptor-α (RXR-α) expression on rat hepatic fibrosis. METHODS: Rat hepatic fibrosis was induced by CCl(4) , and the rats were randomly divided into an early-phase hepatic fibrosis group (2 weeks) and a sustained hepatic fibrosis group (8 weeks). They were then divided into four groups (normal control, hepatic fibrosis, negative control and RXR-α groups). A recombinant lentiviral expression vector carrying the rat RXR-α gene was injected into the rats to induce RXR-α expression by intraportal infusion, hepatic tissue pathological examination was performed, and hydroxyproline content was detected. Hepatic stellate cells (HSC) were cultured in vitro, an RXR-α lentivirus vector was used to activate HSC, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) activation was assayed to detect HSC proliferation. RESULTS: In vivo experiments indicated that in the sustained hepatic fibrosis group, there were significant differences in the hydroxyproline content, and expression of RXR-α, α-smooth muscle actin (α-SMA) and type I collagen (P < 0.01). However, in the early-phase hepatic fibrosis group, hydroxyproline content and the protein level of RXR-α showed no significant difference compared with the normal control group (P > 0.05). In vitro studies revealed that expression of RXR-α significantly inhibited expression of α-SMA and type I collagen in activated HSC (P < 0.01), as well as HSC proliferation (P < 0.01). CONCLUSION: The increased RXR-α gene expression inhibited HSC activation and proliferation and the degree of hepatic fibrosis.
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BACKGROUND: Hepatic fibrosis is a necessary step in the development of hepatic cirrhosis. In this study we used lentiviral vector-mediated transfection technology to evaluate the effect of peroxisome proliferator-activated receptor gamma (PPAR-gamma) on rat hepatic fibrosis. METHODS: Hepatic fibrosis in rats was induced by CCl4 for 2 weeks (early fibrosis) and 8 weeks (sustained fibrosis). The rats were randomly divided into four groups: normal control, fibrosis, blank vector, and PPAR-gamma. They were infected with the recombinant lentiviral expression vector carrying the rat PPAR-gamma gene by portal vein injection. The liver of the rats was examined histologically and hydroxyproline was assessed. In vitro primary hepatic stellate cells (HSCs) were infected with the recombinant lentiviral expression vector carrying the rat PPAR-gamma gene. The status of HSC proliferation was measured by the MTT assay. The protein levels of PPAR-gamma, alpha-smooth muscle actin (alpha-SMA) and type I collagen expression were evaluated by the Western blotting method. RESULTS: In vitro studies revealed that expression of PPAR-gamma inhibited expression of alpha-SMA and type I collagen in activated HSCs (P<0.01) as well as HSC proliferation (P<0.01). In vivo experiments indicated that in the early hepatic fibrosis group, the hydroxyproline content and the level of collagen I protein in the liver in the PPAR-gamma transfected group were not significantly different compared to the hepatic fibrosis group and the blank vector group; whereas the expressions of PPAR-gamma and alpha-SMA were different compared to the hepatic fibrosis group (P<0.01). In the sustained hepatic fibrosis group, there were significant differences in the hydroxyproline content and the expression of PPAR-gamma, alpha-SMA, and type I collagen between each group. CONCLUSION: PPAR-gamma can inhibit HSC proliferation and hepatic fibrosis, and suppress alpha-SMA and type I collagen expression.