Cyclopeptide moroidin inhibits vasculogenic mimicry formed by glioblastoma cells via regulating β-catenin activation and EMT pathways

Glioblastoma(GBM)is a highly vascularized malignant brain tumor with poor clinical outcomes.Vasculogenic mimicry(VM)formed by aggressive GBM cells is an alternative approach for tumor blood supply and contributes to the failure of anti-angiogenic therapy.To date,there is still a lack of effective drugs that target VM formation in GBM.In the present study,we evaluated the effects of the plant cyclopeptide moroidin on VM formed by GBM cells and investigated its underlying molecular mechanisms.Moroidin significantly suppressed cell migration,tube formation,and the expression levels ofα-smooth muscle actin and matrix metalloproteinase-9 in human GBM cell lines at sublethal concentrations.The RNA sequencing data suggested the involvement of the epithelialmesenchymal transition(EMT)pathway in the mechanism of moroidin.Exposure to moroidin led to a concentration-dependent decrease in the expression levels of the EMT markers N-cadherin and vimentin in GBM cells.Moreover,moroidin significantly reduced the level of phosphorylated extracellular signal-regulated protein kinase(p-ERK)and inhibited the activation of β-catenin.Finally,we demonstrated that the plant cyclopeptide moroidin inhibited VM formation by GBM cells through inhibiting the ERK/β-catenin-mediated EMT.Therefore,our study indicates a potential application of moroidin as an anti-VM agent in the treatment of GBM.

Genetic analysis and candidate gene identification of salt tolerancerelated traits in maize

Soil salinization poses a threat to maize production worldwide,but the genetic mechanism of salt tolerance in maize is not well understood.Therefore,identifying the genetic components underlying salt tolerance in maize is of great importance.In the current study,a teosinte-maize BC2F7 population was used to investigate the genetic basis of 21 salt tolerance-related traits.In total,125 QTLs were detected using a high-density genetic bin map,with one to five QTLs explaining 6.05–32.02%of the phenotypic variation for each trait.The total phenotypic variation explained(PVE)by all detected QTLs ranged from 6.84 to 63.88%for each trait.Of all 125 QTLs,only three were major QTLs distributed in two genomic regions on chromosome 6,which were involved in three salt tolerance-related traits.In addition,10 pairs of epistatic QTLs with additive effects were detected for eight traits,explaining 0.9 to 4.44%of the phenotypic variation.Furthermore,18 QTL hotspots affecting 3–7 traits were identified.In one hotspot(L5),a gene cluster consisting of four genes(ZmNSA1,SAG6,ZmCLCg,and ZmHKT1;2)was found,suggesting the involvement of multiple pleiotropic genes.Finally,two important candidate genes,Zm00001d002090 and Zm00001d002391,were found to be associated with salt tolerance-related traits by a combination of linkage and marker-trait association analyses.Zm00001d002090 encodes a calcium-dependent lipid-binding(CaLB domain)family protein,which may function as a Ca^(2+)sensor for transmitting the salt stress signal downstream,while Zm00001d002391 encodes a ubiquitin-specific protease belonging to the C19-related subfamily.Our findings provide valuable insights into the genetic basis of salt tolerance-related traits in maize and a theoretical foundation for breeders to develop enhanced salt-tolerant maize varieties.

Random Green's Function Method for Large-Scale Electronic Structure Calculation

We report a linear-scaling random Green's function(rGF) method for large-scale electronic structure calculation. In this method, the rGF is defined on a set of random states and is efficiently calculated by projecting onto Krylov subspace. With the rGF method, the Fermi–Dirac operator can be obtained directly, avoiding the polynomial expansion to Fermi–Dirac function. To demonstrate the applicability, we implement the rGF method with the density-functional tight-binding method. It is shown that the Krylov subspace can maintain at small size for materials with different gaps at zero temperature, including H_(2)O and Si clusters. We find with a simple deflation technique that the rGF self-consistent calculation of H_(2)O clusters at T = 0 K can reach an error of~ 1 me V per H_(2)O molecule in total energy, compared to deterministic calculations. The rGF method provides an effective stochastic method for large-scale electronic structure simulation.

Generation and characterization of an anti-GP73 monoclonal antibody for immunoblotting and sandwich ELISA

Recently, serum Golgi protein 73 （GP73） levels have been found to be elevated in patients with hepatocellu- lar carcinoma （HCC）, and GP73 has been proposed as a novel marker for HCC. However, GP73 levels in patients remain controversial due to the specificity of the anti-GP73 antibody-based enzyme linked immunosorbent as- say （ELISA）. Therefore, an anti-GP73 antibody with high specificity was highly demanded. In the present study, by hybridoma screening, we generated an anti-GP73 monoclonal antibody （mAb） designated as 6A2 using recom- binant GP73 protein produced by prokaryotic expression. The specificity of 6A2 was evaluated by Western blot- ting, immunohistochemistry and immunoprecipitation. The results showed that 6A2 recognized GP73 in both native and denatured forms. In addition, we have developed a sandwich ELISA using 6A2 and GP73 polyclonal antibody generated in New Zealand white rabbits according to standard procedures, and measured the serum GP73 level of patients using this assay. Our results showed that serum GP73 levels of HCC patients were significantly higher than those of healthy controls （P = 0.0036）. Furthermore, for the first time, GP73 serum level was found to be elevated in patients with breast cancer compared with healthy controls （P = 0.0172）.

Reversing effects of traditional Chinese antitumor medicines on colorectal tumor immunosuppression of natural killer cell and T lymphocyte in vitro

Objective： Reversing effects of traditional Chinese medicines on colorectal tumor immunosuppressions of natural killer （NK） cell and T lymphocyte were analyzed to provide evidence on selecting medicines for patients according to the differ- ent types of tumor immunosuppression. Methods： Six traditional Chinese medicines, including Arsenious acid （AS）, Ligustra- zine hydrochloride （LHC）, Astragalus mongholicus bge （AMB）, Matrine N-oxide （MOX）, Polyporus umbellatus polysaccharide （PUPS） and Artesunate （ART）, were enrolled. The reversing effects on suppression of murine splenocyte transformation and NK killing activity were measured by 3-{4,5-dimethyl-2-thiazolyl}-2,5-diphenyl tetrazolium （MTT）, and the effects on the suppressed expression of intefieukin 2 receptor a （IL-2R（I）, CD3E＊~,＊ and CD3~.-~＊ were detected by flow cytometry （FCM）. The effects on immunosuppressive molecules were measured by enzyme linked immunosorbent assay （ELISA）, including transforming growth factor ~1 （TGF-~I）, vascular endothelial growth factor （VEGF）, interleukin 4 （IL-4）, IL-6, IL-10 and pros- taglandin （PG） E2. Results： （1） The reversing effects of AMB on the inhibition of NK killing and CD3 expression were the most significant; the effect of LHC on inhibition of CD3 expression was the strongest; the effects of AMB, PUPS and ART on inhibition of transformation were the greatest; and the effect of ART on inhibition of IL-2Ra expression was the strongest. （2） The correlated molecules of these medicines that exerted reversing effects on colorectal tumor immunosuppression were TGF-~I and IL-10. AMB had the highest down-regulating effect on the secretion of TGF-~I. AS and ART had the highest effects on IL-10. Conclusion： Reversing tumor immunosuppression through the down-regulation of immunosuppressive molecules is one of the novel antitumor mechanisms of traditional Chinese medicines. The clinical use of compounded prescriptions of ART combined with AMB and LHC should be considered to avoid the reduced treatment efficiency caused by tumor im- munosuppression.

Programmable electron density patterns induced by the interaction of an array laser and underdense plasma

The spatially modulated electron distribution of plasma is the basis for obtaining programmable electron density patterns.It has an important influence on plasma technology applications.We propose an efficient scheme to realize controllable electron density patterns in underdense plasma based on the array laser-plasma interaction.Theoretical evidence for the realization of programmable electron density patterns and the corresponding electrostatic field is provided analytically,which is confirmed by particle-in-cell simulations.Results show that the spatial distribution of electron density in the propagation and transverse directions of the laser can be highly modulated to obtain rich programmable electron density patterns by adjusting the array pattern code and pulse width of the array laser beam.

Dynamics and functionalities of bacterial community during foxtail millet dough fermentation by metagenomic analysis

Sourdough flavors were closely related to microbial metabolism.The microbial diversity of foxtail millet dough during fermentation has never been studied.Here,the metabolic potential and diversity of the bactenial community were analyzed by metagenomic during dough fermentation.Firmicutes was the dominant phylum in the dough,within heterofementative lactic acid bacteria(e.g.Companilactobacillus,Limosilactobacillus,Pediococcus and Lactobacillus)as the most abundant bacteria.Proteobacteria was gradually inhibited after fermentation.Companilactobacillus_crustorum was notably found abundant during dough 1eavening.Limosilactobacillus_fermentum increased markedly during fermentation,while Companilactobacillus_cnustorum decreased significantly.For futher exploration,genes associated with metabolism were annotated through metagenomics.Limosilactobacillus,Companilactobacillus and Pediococcus were actively engaged in glycolysis(ko00010),starch and sucrose metabolism(ko00500),and pyruvate metabolism(ko00620),leading in part to 1actic and acetic acid acumulations and dough acidification.Limosilactobacillus and Lactiplantibacillus were the main contributors to key aninopeptidases ox/and transaminases involved in amino acid metabolism,which was nesponsible for flavor metabolite formation.This study will provide an enhanced understanding of the predominance and diversity of dough bactenial communities,and contribute to future strain screening in the dough for better flavor.

A Regularized Singular Boundary Method for Inverse Cauchy Problem in Three-Dimensional Elastostatics

The application of the singular boundary method(SBM),a relatively new meshless boundary collocation method,to the inverse Cauchy problem in threedimensional(3D)linear elasticity is investigated.The SBM involves a coupling between the non-singular boundary element method(BEM)and the method of fundamental solutions(MFS).The main idea is to fully inherit the dimensionality advantages of the BEM and the meshless and integration-free attributes of the MFS.Due to the boundary-only discretizations and its semi-analytical nature,the method can be viewed as an ideal candidate for the solution of inverse problems.The resulting ill-conditioned algebraic equations is regularized here by employing the first-order Tikhonov regularization technique,while the optimal regularization parameter is determined by the L-curve criterion.Numerical results with both smooth and piecewise smooth geometries show that accurate and stable solution can be obtained with a comparatively large level of noise added into the input data.