Antifungal efficacy of Bacillus amyloliquefaciens ZK-9 against Fusarium graminearum and analysis of the potential mechanism of its lipopeptides.

Fusarium graminearum is a destructive fungal pathogen that seriously threatens wheat production and quality. In the management of fungal infections, biological control is an environmentally friendly and sustainable approach. Here, the antagonistic strain ZK-9 with a broad antifungal activity was identified as Bacillus amyloliquefaciens. ZK-9 could produce extracellular enzymes such as pectinase, protease, cellulase, and amylase, as well as plant growth-promoting substances including IAA and siderophore. Lipopeptides extracted from strain ZK-9 had the high inhibitory effects on the mycelia of F. graminearum with the minimum inhibitory concentration (MIC) of 0.8 mg/mL. Investigation on the action mechanism of lipopeptides showed they could change the morphology of mycelia, damage the cell membrane, lower the content of ergosterol and increase the relative conductivity of membrane, cause nucleic acid and proteins leaking out from the cells, and disrupt the cell membrane permeability. Furthermore, metabolomic analysis of F. graminearum revealed the significant differences in the expression of 100 metabolites between the lipopeptides treatment group and the control group, which were associated with various metabolic pathways, mainly including amino acid biosynthesis, pentose, glucuronate and glycerophospholipid metabolism. In addition, strain ZK-9 inhibited Fusarium crown rot (FCR) with a biocontrol efficacy of 82.14 % and increased the plant height and root length by 24.23 % and 93.25 %, respectively. Moreover, the field control efficacy of strain ZK-9 on Fusarium head blight (FHB) was 71.76 %, and the DON content in wheat grains was significantly reduced by 69.9 %. This study puts valuable insights into the antifungal mechanism of lipopeptides against F. graminearum, and provides a promising biocontrol agent for controlling F. graminearum.

RGBT Tracking via Challenge-Based Appearance Disentanglement and Interaction.

RGB and thermal source data suffer from both shared and specific challenges, and how to explore and exploit them plays a critical role in representing the target appearance in RGBT tracking. In this paper, we propose a novel approach, which performs target appearance representation disentanglement and interaction via both modality-shared and modality-specific challenge attributes, for robust RGBT tracking. In particular, we disentangle the target appearance representations via five challenge-based branches with different structures according to their properties, including three parameter-shared branches to model modality-shared challenges and two parameter-independent branches to model modality-specific challenges. Considering the complementary advantages between modality-specific cues, we propose a guidance interaction module to transfer discriminative features from one modality to another one to enhance the discriminative ability of weak modality. Moreover, we design an aggregation interaction module to combine all challenge-based target representations, which could form more discriminative target representations and fit the challenge-agnostic tracking process. These challenge-based branches are able to model the target appearance under certain challenges so that the target representations can be learned by a few parameters even in the situation of insufficient training data. In addition, to relieve labor costs and avoid label ambiguity, we design a generation strategy to generate training data with different challenge attributes. Comprehensive experiments demonstrate the superiority of the proposed tracker against the state-of-the-art methods on four benchmark datasets.

SREBP2 inhibitor betulin sensitizes hepatocellular carcinoma to lenvatinib by inhibiting the mTOR/IL-1ß pathway.

Lenvatinib has become the first-line therapy in advanced hepatocellular carcinoma (HCC), but its efficacy is still limited because of the inevitable development of resistance. It has been reported that cellular cholesterol levels are associated with tyrosine kinase inhibitor (TKI) efficacy. Here, we show that betulin, a sterol regulatory element-binding protein 2 (SREBP2) inhibitor, markedly enhances the anti-tumor effect of lenvatinib in HCC both in vitro and in vivo. Our results also show that the combination treatment of lenvatinib and betulin synergistically inhibits the proliferation and clonogenicity of HCC cells. The mRNA and protein expressions of IL-1ß are markedly decreased in HCC cells treated with betulin, while the sensitivity of HCC cells to lenvatinib is enhanced. Moreover, we find that the knockdown of IL-1ß also enhances the efficacy of lenvatinib, and recombinant IL-1ß protein rescues cell viability, which is reduced by lenvatinib in HCC cells. Further mechanistic studies indicate that betulin decreases the level of IL-1ß in HCC cells by inhibiting the mTOR signaling pathway. Finally, the growth of the tumors in xenograft mouse models subjected to combination treatment is significantly suppressed. In summary, our study reveals that the SREBP2 inhibitor betulin sensitizes hepatocellular carcinoma to lenvatinib by inhibiting the mTOR/IL-1ß pathway, which may be a promising therapeutic strategy for patients with HCC.

In Vivo Assessment of the Effects of Mono-Carrier Encapsulated Fucoxanthin Nanoparticles on Type 2 Diabetic C57 Mice and Their Oxidative Stress.

Fucoxanthin (FX) is a carotenoid from a marine origin that has an important role in our health, especially in the regulation and alleviation of type 2 diabetes. Its specific molecular structure makes it very unstable, which greatly affects its delivery in the body. In this study, FX was encapsulated in a mono-carrier using a hydrolyzed zein to form a nanocomplex with a stable structure and chemical properties (FZNP). Its stability was demonstrated by characterization and the efficacy of FX before and after encapsulation in alleviating diabetes in mice, which was evaluated by in vivo experiments. FZNP reduced the level of fasting blood glucose and restored it to normal levels in T2DM mice, which was not caused by a decrease in food intake, and effectively reduced oxidative stress in the organism. Both FX and FZNP repaired the hepatocyte and pancreatic ß-cell damage, increased serum SOD and reduced INS values significantly, upregulated PI3K-AKT genes as well as CaMK and GNAs expression in the pancreas. FZNP increased ADPN and GSH-PX values more significantly and it decreased serum HOMA-IR and MDA values, upregulated GLUT2 expression, promoted glucose transport in pancreatic and hepatocytes, regulated glucose metabolism and glycogen synthesis with much superior effects than FX.

Mn(II)-Mediated Self-Assembly of Tea Polysaccharide Nanoparticles and Their Functional Role in Mice with Type 2 Diabetes.

Tea polysaccharide (TPS) is a bioactive compound that has attracted increasing attention for its health effect on regulating the metabolism of glucose and lipid. Moreover, due to their good biocompatibility and biodegradability, TPS-based nanoparticles have emerged as effective nanocarriers for the delivery of bioactive molecules. In this study, we developed a TPS-based biocarrier system for the orally targeted administration of Mn(II) ions and investigated their antidiabetic effects in C57BL/6 mice with HFD/streptozotocin (STZ)-induced T2DM. Mn(II)-loaded TPS-based nanoparticles (MTNPs) were synthesized, in which negatively charged functional groups in protein and uronic acid in TPS conjugates would act as binding sites for Mn(II) ions, which is responsible for the cross-linking reaction of MTNP. The resulting MTNP had a spherical shape and a mean particle size of around 30 nm with a Mn(II) ion content of 2.24 ± 0.13 mg/g. In T2DM mice, we discovered that MTNP treatment significantly lowered blood glucose levels and improved glucose intolerance. Furthermore, the impact of MTNP on the recovery of FINS, the homeostatic index of insulin resistance (HOMA-IR), and the homeostatic index of ß-cell (HOMA ß-cell) levels was significantly larger (p < 0.05) than TPS alone, demonstrating that Mn(II) ions can enhance TPS's ability to repair HFD/STZ-induced ß-cell damage. Mn(II) ions in MTNP not only acted as cofactors to increase the exocytosis of insulin secretory cells by upregulating the expression of Ca(II)/calmodulin-dependent protein kinase II (CaMK II) but also promoted TPS's lipid-lowering effect in T2DM mice by inhibiting glucogenesis and regulating the lipid metabolism. Our findings suggest that Mn(II) ions can be used not only as cross-linkers in the formation of nanoparticulated TPS but also as cofactors in improving the functional role of TPS in regulating the glucose and lipid metabolism, which will provide insights into the development of TPS-based drug delivery systems for the prevention of type 2 diabetes.

Cholesterol suppresses GOLM1-dependent selective autophagy of RTKs in hepatocellular carcinoma.

Aberrant activation of receptor tyrosine kinases (RTKs) and the subsequent metabolic reprogramming play critical roles in cancer progression. Our previous study has shown that Golgi membrane protein 1 (GOLM1) promotes hepatocellular carcinoma (HCC) metastasis by enhancing the recycling of RTKs. However, how this RTK recycling process is regulated and coupled with RTK degradation remains poorly defined. Here, we demonstrate that cholesterol suppresses the autophagic degradation of RTKs in a GOLM1-dependent manner. Further mechanistic studies reveal that GOLM1 mediates the selective autophagy of RTKs by interacting with LC3 through an LC3-interacting region (LIR), which is regulated by a cholesterol-mTORC1 axis. Lowering cholesterol by statins improves the efficacy of multiple tyrosine kinase inhibitors (TKIs) in vivo. Our findings indicate that cholesterol serves as a signal to switch GOLM1-RTK degradation to GOLM1-RTK recycling and suggest that lowering cholesterol by statin may be a promising combination strategy to improve the TKI efficiency in HCC.

GOLM1 exacerbates CD8+ T cell suppression in hepatocellular carcinoma by promoting exosomal PD-L1 transport into tumor-associated macrophages.

The immunosuppressive microenvironment plays an important role in tumor progression and immunotherapy responses. Golgi membrane protein 1 (GOLM1) is correlated to hepatocellular carcinoma (HCC) progression and metastasis. However, little is known about the role of GOLM1 in regulating the immunosuppressive environment and its impact on immunotherapeutic efficacy in HCC. In this study, GOLM1 was positively correlated with infiltrating tumor-associated macrophages (TAMs) expressed high levels of programmed death-ligand 1 (PD-L1) and CD8+ T cell suppression in HCC tissues. Both gain- and loss-of-function studies determined a close correlation between GOLM1 and immunosuppression. In the mechanism, GOLM1 promoted COP9 signalosome 5-mediated PD-L1 deubiquitination in HCC cells and increased the transport of PD-L1 into exosomes via suppression of Rab27b expression. Furthermore, co-culture with exosomes derived from HCC cells upregulated the expression of PD-L1 on macrophages. Zoledronic acid in combination with anti-PD-L1 therapy reduced PD-L1+ TAMs infiltration and alleviated CD8+ T cell suppression, resulting in tumor growth inhibition in the mouse HCC model. Together, our study unveils a mechanism by which GOLM1 induces CD8+ T cells suppression through promoting PD-L1 stabilization and transporting PD-L1 into TAMs with exosome dependent. Targeting PD-L1+ TAM could be a novel strategy to enhance the efficacy of anti-PD-L1 therapy in HCC.

Preparation and characterization of metal-tea polysaccharide complexes and their inhibition on α-glucosidase.

The preparation method and the sources of metal elements may affect the activity of the metal-polysaccharide complex. In this study, four Fe-tea polysaccharide complexes were prepared and three tea polysaccharides (TPSs) from different seasons were extracted. Moreover, the binding mode of TPSs with internal and external metallic elements as well as their inhibitory effect on α-glucosidase was explored. The results revealed that the binding mode (-C-O-Fe and -C-Fe) of the Fe-TPS complex prepared at pH 5.0 was closer to TPS with internal metallic elements. The TPS with the least amount of internal metallic elements (61.72 mg/g) exhibited a high inhibitory activity on α-glucosidase (37.90%). The inhibitory activity of Fe-TPS on α-glucosidase was lower than that without Fe. But the quenching effect and the inhibition type of TPSs on α-glucosidase were not affected by metallic elements. Therefore, the metallic elements have the potential to reduce the hypoglycemic activity of TPS. PRACTICAL APPLICATIONS: In this paper, TPS was extracted from crude tea in different seasons, and the effects of metallic elements in TPS on hypoglycemic activity, physicochemical properties, and structure of TPS were discussed. TPS metal complexes were prepared by adding Fe3+ or removing metallic elements, and the differences of internal metallic elements in TPS were discussed. It is of great academic significance to use tea pruned leaves and crude tea as potential resources to develop polysaccharide hypoglycemic products and to reveal the relationship between TPS metal ions and their structure and activity. In addition, it has guiding value for consumers to choose tea-producing regions and growers to choose chemical fertilizer.

Effect of purity of tea polysaccharides on its antioxidant and hypoglycemic activities.

The effects of purity of tea polysaccharides (TPS) on its five antioxidant activities and hypoglycemic activities in vitro were studied. The results showed that the higher the purity of TPS, the lower the antioxidant capacity. The purity of FTPSI is the highest (sugar content 80.72%), but its antioxidant activities were lower than those of Fujian tea polysaccharides (FTPS) and FTPSII. The antioxidant activity of tea polysaccharide is related to its protein and polyphenol content (Pearson r > .90). The protective effect of Zhejiang tea polysaccharides and FTPS on human umbilical vein endothelial cells (HUVEC) was better than that of its purified fractions. The inhibition rates of FTPSII (5 and 2 mg/ml) on α-glucosidase (32.76%) and α-amylase (-11.93%) were higher than those of FTPS and FTPSII. Purification does not change the basic structure of TPS. This study has certain reference value for the study of the antioxidant activities of TPS. Meanwhile, TPS can be used as a potential resource with hypoglycemic function. PRACTICAL APPLICATIONS: A large number of studies have shown that TPS have antioxidant activity. However, several studies considered that the antioxidant activity of TPS mainly comes from the residues of tea polyphenols. Therefore, the in vitro and cell antioxidant activities of TPS were studied in this paper. We believe that both glycoprotein and tea polyphenol are antioxidants of tea, and tea polysaccharide perform preferable effect on hypoglycemic. HUVEC cell model and four in vitro antioxidant test methods were used to study the antioxidant activities of TPS, and two enzyme inhibition activities were used to study the hypoglycemic effect of TPS, in order to provide a theoretical basis for the study of biological activity of TPS.

Selective, highly efficient extraction of Cr(III), Pb(II) and Fe(III) from complex water environment with a tea residue derived porous gel adsorbent.

A novel macroporous (~150 µm) double network hydrogel (TR/PAA) was prepared from tea residue and acrylic acid, and its performance was systematically evaluated. The static adsorption experiments showed that gel exhibited high selectivity and adsorption capacity, ultrafast kinetics (~10 min) for Cr(III), Pb(II) and Fe(III). The adsorption behavior showed heterogeneous and chemisorption process adsorption capacities of 206.19, 253.16, and 94.88 mg g-1 for Cr(III), Pb(II) and Fe(III), respectively. In pluralistic systems, TR/PAA showed the adsorption order of Fe(III) > Cr(III) > Pb(II). Mechanism studies confirm that nitrogen and oxygen-containing functional groups play a major role in the adsorption process. In the fixed-bed column experiments, the treatment volume of simulated wastewater reached 1400 bed volumes (BV) (21.6 L), producing only 7 BV (323 mL) eluent. This work provides a new avenue for the combination of TR/PAA reuse and heavy metal removal, which is expected to be applied in actual wastewater treatment.

Fabrication of stable zein nanoparticles by chondroitin sulfate deposition based on antisolvent precipitation method.

The purpose of this work is to stabilize zein nanoparticles with anionic polysaccharides-chondroitin sulfate (CS) to overcome the poor colloidal stability of zein nanoparticles. The average molecular weight of CS was 20.51 kDa and the disaccharide composition of CS was 70.71% monosulfated disaccharide Di2S and 29.29% disulfated disaccharide Di2,6 diS. Zein/CS composite nanoparticles were fabricated by antisolvent precipitation method at pH 4.0. The optimal mass ratio of zein to CS was 1:1 (w/w). Zein/CS composite nanoparticles showed mean size of 148 nm and PDI < 0.2, maintaining great stability at pH 3.0 to 8.0. Meanwhile, these prepared nanoparticles were also stable to heat treatment and was stable to a range of ionic strength of 0-15 mM at pH 4.0 and 0-10 mM at pH 7.0. The composite nanoparticles exhibited regular spherical structures, and CS was deposited on the surface of zein mainly by electrostatic interactions. In the process, the addition of urea and sodium dodecyl sulfate indicated that it also involved both hydrogen bonding and hydrophobic interactions. In addition, cytotoxicity assay on human renal epithelial cells-293 cells indicated that zein/CS composite nanoparticles were essentially nontoxic.

Effects of intrinsic metal ions of lentinan with different molecular weights from Lentinus edodes on the antioxidant capacity and activity against proliferation of cancer cells.

In this study, effects of intrinsic metal ions in lentinan on its bioactivities were evaluated. Lentinan extracted from Lentinus edodes was separated to four fractions with different molecular weights with Sephadex G-200 gel chromatography, and intrinsic metal ions were removed by EDTA. The ferrous ion chelating capacity of ELWs was measured to estimate EDTA residue after extensive dialysis. Antioxidant and antitumor activities of the isolated lentinans (LWs and ELWs) were investigated in vitro and compared, respectively. The results indicated that all of the isolated lentinan contained a large amount of intrinsic metal ions and the fraction with the lowest Mw had highest metal content. After removing intrinsic metal ions, the polysaccharides showed lower ABTS and DPPH radicals scavenging capacity and the reduced inhibition of the proliferation of BXPC-3 and Hela cells. Our results indicated that the metal ions in lentinan had positive effects on its bioactivities.

Effects of metal ions in tea polysaccharides on their in vitro antioxidant activity and hypoglycemic activity.

Total tea polysaccharides (TTPS) were extracted from two kinds of pruning leaves of tea plant and fractionated into neutral tea polysaccharides (TPSI) and acidic tea polysaccharides (TPSII) by anion exchange resin D315. Some physicochemical properties, including structure, monosaccharide composition, and molecular weight distribution, as well as the 4 in vitro antioxidant activities and inhibitory effects on α-glucosidase of above polysaccharides before and after removing metal ions were investigated. By comparing TTPS and TPSII, we found that they exhibited different antioxidant activities and inhibitory actions against α-glucosidase after their metal ions were removed. However, the in vitro antioxidant activities and inhibitory effects on α-glucosidase of TPSI were substantially improved. The study can be a certain reference for tea and soil selection. At the same time, we suggested that pruning leaves of tea plant could be treated as a potential resource for the development of polysaccharide antioxidants and hypoglycemic products.