Sequential activation of ERα-AMPKα signaling by the flavonoid baicalin down-regulates viral HNF-dependent HBV replication.

Baicalin (BA), a natural component found in many traditional Chinese medicines, exerts protective effects against several viruses. Although our previous studies have revealed that the anti-hepatitis B virus (anti-HBV) activity of BA depends on hepatocyte nuclear factor (HNF) signaling, the specific mechanisms remain unclear. The present study explored the potential signaling mechanisms involved in BA-mediated HBV suppression. Transcriptomic analysis suggested that BA significantly modulates the estrogen receptor (ER) and AMPK signaling pathways in HepG2 cells. The ER alpha (ERα) binding affinity of BA and its estrogen-like agonist activity were subsequently verified through molecular docking assays, BA-ERα affinity detection experiments, ERα luciferase reporter gene assays, and qRT-PCR. ERα knockdown (shRNA) and AMPK inhibition (Compound C and doxorubicin [Dox]) experiments revealed that the sequential activation of the ERα-LKB1-AMPK-HNF signaling axis is essential for the anti-HBV effects of BA. This study indicates that BA may trigger the ERα-AMPKα-HNF pathway to inhibit HBV replication, providing insights into its potential protective mechanisms against other viruses.

Evident structural anisotropies arising from near-zero particle asphericity in granular spherocylinder packings.

With magnetic resonance imaging experiments, we study packings of granular spherocylinders with merely 2% asphericity. Evident structural anisotropies across all length scales are identified. Most interestingly, the global nematic order decreases with increasing packing fraction, while the local contact anisotropy shows an opposing trend. We attribute this counterintuitive phenomenon to a competition between gravity-driven ordering aided by frictional contacts and a geometric frustration effect at the marginally jammed state. It is also surprising to notice that such slight particle asphericity can trigger non-negligible correlations between contact-level and mesoscale structures, manifested in drastically different nonaffine structural rearrangements upon compaction from that of granular spheres. These observations can help improve statistical mechanical models for the orientational order transformation of nonspherical granular particle packings, which involves complex interplays between particle shape, frictional contacts, and external force field.

Intrinsic Stress Field for Liquid Surfaces.

The microscopic stress field inhomogeneity in the interfacial region adjacent to the liquid surface is the fundamental origin of the liquid surface tension, but because of broadening due to capillary fluctuations, a detailed molecular level understanding of the stress field remains elusive. In this work, we deconvolute the capillary fluctuations to reveal the intrinsic stress field and show that the atomic-level contributions to the surface tension are similar in functional form across a variety of monatomic systems. These contributions are confined to an interfacial region approximately 1.5±0.1 times the particle diameter for all systems studied. In addition, the intrinsic density and stress profiles show a strong spatial correlation that should be useful in the development of a statistical mechanical theory for the prediction of surface stress and surface tension.

Approaching Effective Differential Centrifugal Fractionation by Combining Image Analysis with Analytical Ultracentrifugation.

Centrifugation is one of the most commonly used methods for separation in biology and chemistry. However, effective fractionation is not always easy to obtain, as preparative centrifuge experiments are mostly conducted in an empirical way, even when it is guided by the quantitative results from analytical ultracentrifuge (AUC). Very few works have been performed to enhance the fractionation resolution of the differential centrifugation method in a swing-out rotor. This is primarily due to the absence of a characterization tool for sedimentation in the preparative centrifuge. In this study, we utilized image analysis to map the particle concentration distribution throughout the preparative centrifuge tube, revealing an unexpected and abnormal sedimentation process. By characterizing the sedimentation coefficient distributions of the fractionated product via AUC, we demonstrated that the overall sedimentation efficiency in a swing-out preparative centrifuge was significantly reduced. Furthermore, effective fractionation was confined to the intermediate phase of the entire sedimentation process. We propose that the mechanism here is a combination of the inverse Boycott effect and droplet sedimentation. The actual sedimentation process within a preparative centrifuge can be described by modifying the Lamm equation phenomenologically, which simply results in an effective sedimentation coefficient. Our work builds a foundation for determining the optimal preparative centrifugation conditions for various systems.

Structural Transformation between a Nematic Loose Packing and a Randomly Stacked Close Packing of Granular Disks.

Packing structures of granular disks are reconstructed using magnetic resonance imaging techniques. As packing fraction increases, the packing structure transforms from a nematic loose packing to a dense packing with randomly oriented stacks. According to our model based on Edwards' volume ensemble, stack structures are statistically favored when the effective temperature decreases, which has a lower structural anisotropy than single disks, and brings down the global orientational order consequently. This mechanism identified in athermal granular materials can help us understand the nonergodic characteristics of disklike particle assemblies such as discotic mesogens and clays.

Equivalence of Fluctuation-Dissipation and Edwards' Temperature in Cyclically Sheared Granular Systems.

Using particle trajectory data obtained from x-ray tomography, we determine two kinds of effective temperatures in a cyclically sheared granular system. The first one is obtained from the fluctuation-dissipation theorem which relates the diffusion and mobility of lighter tracer particles immersed in the system. The second is the Edwards compactivity defined via the packing volume fluctuations. We find robust agreement between these two temperatures, independent of the type of the tracers, cyclic shear amplitudes, and particle surface roughness, giving therefore the first experimental evidence that the concept of effective temperature is valid in driven frictional granular systems.

Autophagy and Exosome Coordinately Enhance Macrophage M1 Polarization and Recruitment in Influenza A Virus Infection.

Background: Influenza A virus infection results in viral pneumonia, which is often accompanied by the infiltration and recruitment of macrophages, overactivation of inflammatory responses, and obvious cell autophagy and exosome production. However, little is known about the roles of autophagy and exosome production in these inflammatory responses. Methods: In this study, multiple methods, such as flow cytometry, real-time quantitative reverse transcription-polymerase chain reaction, immune-fluorescence technology, and western blot, were applied to explore the possible effects of autophagy and exosome production by H1N1-infected host cells. Results: It was observed that a high number of polarized macrophages (CD11b+/F4/80+/CD86+) were recruited to the lung tissues of infected mice, which could be mimicked by tracking the movement of macrophages to H1N1-infected cells in vitro (transwell assays). Furthermore, there was some coordinated upregulation of M1 polarization signs (iNOS/Arg-1 bias) as well as autophagy (LC3) and exosome (CD63) biomarkers in the infected macrophages and epithelial cells. Moreover, exosomes extracted from the supernatant of virus-infected cells were shown to promote the recruitment and polarization of more peritoneal macrophages than the normal group. The fluorescence colocalization of LC3-CD63 and the inhibition of autophagy and exosome signaling pathway further revealed that H1N1 infection seemed to sequentially activate the M1 polarization and recruitment of macrophages via autophagy-exosome dependent pathway. Conclusion: Autophagy and exosome production coordinately enhance the M1 polarization and recruitment of macrophages in influenza virus infection, which also provides potential therapeutic targets.

Liver-Targeted Nanoparticles Facilitate the Bioavailability and Anti-HBV Efficacy of Baicalin In Vitro and In Vivo.

The anti-hepatitis B virus (HBV) efficacy of baicalin (BA) is mediated by HBV-related hepatocyte nuclear factors (HNFs). However, this efficacy is severely limited by the low bioavailability of BA. Therefore, a novel liver-targeted BA liposome was constructed to promote the bioavailability and antiviral ability of BA. The results showed that apolipoprotein A1 (ApoA1)-modified liposomes (BAA1) significantly enhanced BA's cellular uptake and specific distribution in the liver. Furthermore, the substantial inhibitory effects of BAA1 on HBsAg, HBeAg, HBV RNA, and HBV DNA were assessed in HB-infected cells and mice. Western blotting, co-immunoprecipitation, and transcriptomics analysis further revealed that the enhanced anti-HBV efficacy of BAA1 was attributed to the interaction between hepatocyte nuclear factors (HNFs) and estrogen receptors (ERs). Based on the findings, we propose that the ApoA1-modified liposomes aid BA in inhibiting HBV transcription and replication by augmenting its bioavailability and the HNFs-ERs axis.

Structural evolution of granular cubes packing during shear-induced ordering.

Packings of granular particles may transform into ordered structures under external agitation, which is a special type of out-of-equilibrium self-assembly. Here, evolution of the internal packing structures of granular cubes under cyclic rotating shearing has been analyzed using magnetic resonance imaging techniques. Various order parameters, different types of contacts and clusters composed of face-contacting cubes, as well as the free volume regions in which each cube can move freely have been analyzed systematically to quantify the ordering process and the underlying mechanism of this granular self-assembly. The compaction process is featured by a first rapid formation of orientationally ordered local structures with faceted contacts, followed by further densification driven by free-volume maximization with an almost saturated degree of order. The ordered structures are strongly anisotropic with contacting ordered layers in the vertical direction while remaining liquid-like in the horizontal directions. Therefore, the constraint of mechanical stability for granular packings and the thermodynamic principle of entropy maximization are both effective in this system, which we propose can be reconciled by considering different depths of supercooling associated with various degrees of freedom.

Cubatic structural transformation of the packing of granular cylinders.

Packing structures of granular cylinders with the aspect ratio close to one have been reconstructed with the help of magnetic resonance imaging techniques. By controlling the container boundary conditions and preparation protocols, a structural transformation from a disordered liquid-like state to an orientationally ordered state with cubatic symmetry at a high packing fraction is observed. This ordering process is accompanied by the formation of more faceted contacts, which lower the elastic energy between jammed granular particles to drive the transformation. With the help of Edwards' volume ensemble theory, this granular structural transformation is explained using a phenomenological thermodynamic model and a self-consistent mean-field statistical mechanical model. Both models predict a sharp but continuous change of order parameter when the effective granular temperature is lowered. The intrinsic difference and connection between this granular structural transformation and the entropy-driven phase transition of conventional thermal hard-particle systems are discussed.

Electromagnetic Exposure Dosimetry Study on Two Free Rats at 1.8 GHz via Numerical Simulation.

Normally, the impact of electromagnetic exposure on human health is evaluated by animal study. The biological effect caused by electromagnetic exposure on such experimental animals as rats has been proven to be dose-dependent. However, though the dose of radio frequency (RF) electromagnetic exposure described by the specific absorbing rate (SAR) on fixed rats has been relatively well-studied utilizing the numerical simulations, the dosimetry study of exposure on free rat is insufficient, especially in the cases of two or more free rats. Therefore, the present work focuses on the variation of SAR caused by the existence of neighboring free rat in the same cage. Here, infrared thermography was used to record the activity of the two free rats who lived in the same cage that mounted at the far-field region in the microwave darkroom for a duration of 48 h. Then, using image processing techniques, the relative positions and orientations of the two rats are identified, which are defined by three parameters, such as the relative distance (d), relative direction angle (α), and relative orientation angle (ß). Using the simulation software XFdtd 7.3, the influence of d, α, and ß on the whole-body average SAR (WB-avgSAR) of the rats exposed to 1.8 GHz electromagnetic wave was calculated and analyzed. Then, the average variation of WB-avgSAR of the two rats compared with that of a single rat within 48 h was calculated. The numerical simulation results showed that the relative posture position described by (d, α, and ß) of the two rats affects their WB-avgSAR and leads to fluctuations at different positions. However, the variation rate of the 48-h-average WB-avgSAR was only 10.3%, which implied that the over-time average SAR of two or more rats can be roughly described by the WB-avgSAR of a single free rat, except when a real-time precise control of exposure dose is necessary.

Experimental Test of the Edwards Volume Ensemble for Tapped Granular Packings.

Using x-ray tomography, we experimentally investigate granular packings subject to mechanical tapping for three types of beads with different friction coefficients. We validate the Edwards volume ensemble in these three-dimensional granular systems and establish a granular version of thermodynamic zeroth law. Within the Edwards framework, we also explicitly clarify how friction influences granular statistical mechanics by modifying the density of states, which allows us to determine the entropy as a function of packing fraction and friction. Additionally, we obtain a granular jamming phase diagram based on geometric coordination number and packing fraction.

Structural characterization and statistical properties of jammed soft ellipsoid packing.

The jamming transition and jammed packing structures of hydrogel soft ellipsoids are studied using magnetic resonance imaging techniques. As the packing fraction increases, the fluctuation of local free volume decreases and the fluctuation of particle deformation increases. Effective thermodynamic quantities are obtained by characterizing these fluctuations using k-gamma distributions based on an underlying statistical model for granular materials. Surprisingly, the two granular temperatures measuring the relative fluctuations of both free volume and particle deformation remain basically unchanged as the packing fraction increases. The total configurational entropy is also approximately constant for packing with different packing fractions. The significantly different behaviors of these effective thermodynamic quantities compared with hard sphere systems are further attributed to a statistically affine structural transformation of the packing structures along with particle deformations when the packing fraction changes.

Baicalin Inhibits Influenza A Virus Infection via Promotion of M1 Macrophage Polarization.

BACKGROUND AND AIMS: The natural compound baicalin (BA) possesses potent antiviral properties against the influenza virus. However, the underlying molecular mechanisms of this antiviral activity and whether macrophages are involved remain unclear. In this study, we, therefore, investigated the effect of BA on macrophages. METHODS: We studied macrophage recruitment, functional phenotypes (M1/M2), and the cellular metabolism via flow cytometry, qRT-PCR, immunofluorescence, a cell culture transwell system, and GC-MS-based metabolomics both in vivo in H1N1 A virus-infected mice and in vitro. RESULTS: BA treatment drastically reduced macrophage recruitment (CD11b+, F4/80+) by approximately 90% while maintaining the proportion of M1-polarized macrophages in the bronchoalveolar lavage fluid of infected mice. This BA-stimulated macrophage M1 phenotype shift was further verified in vitro in ANA-1 and primary peritoneal macrophages by measuring macrophage M1 polarization signals (CD86, iNOS, TNF-α, iNOS/Arg-1 ratio, and IL-1ß cleavage). Meanwhile, we observed an activation of the IFN pathway (upregulation of IFN-ß and IRF-3), an inhibition of influenza virus replication (as measured by the M gene), and distinct cellular metabolic responses in BA-treated cells. CONCLUSION: BA triggered macrophage M1 polarization, IFN activation, and other cellular reactions, which are beneficial for inhibition of H1N1 A virus infection.

Baicalin down-regulating hepatitis B virus transcription depends on the liver-specific HNF4α-HNF1α axis.

Baicalin (BA) inhibits hepatitis B virus (HBV) RNAs production and reduces levels of the related hepatocyte nuclear factors (HNFs), although the underlying mechanism is unclear. In this study, we investigated the specific pathway by which BA regulates HBV transcription through the HBV-related HNFs. Following transfection of HepG2 cells with pHBV1.2, we observed that BA inhibited the production of HBV RNAs and viral proteins in a time- and dose-dependent manner. These effects were consistent with the downregulation of HNF1α, which was abolished by HNF1α-shRNA. The shRNA of HNF4α, the upstream gene of HNF1α, also remarkedly reduced HNF1α expression and impaired the anti-HBV efficacy of BA, indicating that this function of BA depended on HNF4α/HNF1α axis. Furthermore, chromatin immunoprecipitation assay showed that BA significantly reduced HNF4α-HNF1α transactivation activity. The similar effects of BA were observed in entecavir (ETV)-resistant HBVrtM204V/rtLl80M transfected HepG2 cells. Thus, we proposed a mechanism for the anti-HBV activity of BA in an HNF4α-HNF1α-dependent manner, which impaired HNF4α and HNF1α transactivation, and effectively inhibited HBV transcription and viral replication.

Translational and Rotational Dynamical Heterogeneities in Granular Systems.

We use x-ray tomography to investigate the translational and rotational dynamical heterogeneities of a three dimensional hard ellipsoid granular packing driven by oscillatory shear. We find that particles which translate quickly form clusters with a size distribution given by a power law with an exponent that is independent of the strain amplitude. Identical behavior is found for particles that are translating slowly, rotating quickly, or rotating slowly. The geometrical properties of these four different types of clusters are the same as those of random clusters. Different cluster types are considerably correlated or anticorrelated, indicating a significant coupling between translational and rotational degrees of freedom. Surprisingly, these clusters are formed already at time scales that are much shorter than the α-relaxation time, in stark contrast to the behavior found in glass-forming systems.

Structural and topological nature of plasticity in sheared granular materials.

Upon mechanical loading, granular materials yield and undergo plastic deformation. The nature of plastic deformation is essential for the development of the macroscopic constitutive models and the understanding of shear band formation. However, we still do not fully understand the microscopic nature of plastic deformation in disordered granular materials. Here we used synchrotron X-ray tomography technique to track the structural evolutions of three-dimensional granular materials under shear. We establish that highly distorted coplanar tetrahedra are the structural defects responsible for microscopic plasticity in disordered granular packings. The elementary plastic events occur through flip events which correspond to a neighbor switching process among these coplanar tetrahedra (or equivalently as the rotation motion of 4-ring disclinations). These events are discrete in space and possess specific orientations with the principal stress direction.

Granular materials flow like complex fluids.

Granular materials such as sand, powders and foams are ubiquitous in daily life and in industrial and geotechnical applications. These disordered systems form stable structures when unperturbed, but in the presence of external influences such as tapping or shear they 'relax', becoming fluid in nature. It is often assumed that the relaxation dynamics of granular systems is similar to that of thermal glass-forming systems. However, so far it has not been possible to determine experimentally the dynamic properties of three-dimensional granular systems at the particle level. This lack of experimental data, combined with the fact that the motion of granular particles involves friction (whereas the motion of particles in thermal glass-forming systems does not), means that an accurate description of the relaxation dynamics of granular materials is lacking. Here we use X-ray tomography to determine the microscale relaxation dynamics of hard granular ellipsoids subject to an oscillatory shear. We find that the distribution of the displacements of the ellipsoids is well described by a Gumbel law (which is similar to a Gaussian distribution for small displacements but has a heavier tail for larger displacements), with a shape parameter that is independent of the amplitude of the shear strain and of the time. Despite this universality, the mean squared displacement of an individual ellipsoid follows a power law as a function of time, with an exponent that does depend on the strain amplitude and time. We argue that these results are related to microscale relaxation mechanisms that involve friction and memory effects (whereby the motion of an ellipsoid at a given point in time depends on its previous motion). Our observations demonstrate that, at the particle level, the dynamic behaviour of granular systems is qualitatively different from that of thermal glass-forming systems, and is instead more similar to that of complex fluids. We conclude that granular materials can relax even when the driving strain is weak.

Origin of Noncubic Scaling Law in Disordered Granular Packing.

Recent diffraction experiments on metallic glasses have unveiled an unexpected noncubic scaling law between density and average interatomic distance, which led to the speculation of the presence of fractal glass order. Using x-ray tomography we identify here a similar noncubic scaling law in disordered granular packing of spherical particles. We find that the scaling law is directly related to the contact neighbors within the first nearest neighbor shell, and, therefore, is closely connected to the phenomenon of jamming. The seemingly universal scaling exponent around 2.5 arises due to the isostatic condition with a contact number around 6, and we argue that the exponent should not be universal.

The structural origin of the hard-sphere glass transition in granular packing.

Glass transition is accompanied by a rapid growth of the structural relaxation time and a concomitant decrease of configurational entropy. It remains unclear whether the transition has a thermodynamic origin, and whether the dynamic arrest is associated with the growth of a certain static order. Using granular packing as a model hard-sphere glass, we show the glass transition as a thermodynamic phase transition with a 'hidden' polytetrahedral order. This polytetrahedral order is spatially correlated with the slow dynamics. It is geometrically frustrated and has a peculiar fractal dimension. Additionally, as the packing fraction increases, its growth follows an entropy-driven nucleation process, similar to that of the random first-order transition theory. Our study essentially identifies a long-sought-after structural glass order in hard-sphere glasses.