Molecular modeling and simulation approaches to characterize potential molecular targets for burdock inulin to instigate protection against autoimmune diseases.

In the current study, we utilized molecular modeling and simulation approaches to define putative potential molecular targets for Burdock Inulin, including inflammatory proteins such as iNOS, COX-2, TNF-alpha, IL-6, and IL-1ß. Molecular docking results revealed potential interactions and good binding affinity for these targets; however, IL-1ß, COX-2, and iNOS were identified as the best targets for Inulin. Molecular simulation-based stability assessment demonstrated that inulin could primarily target iNOS and may also supplementarily target COX-2 and IL-1ß during DSS-induced colitis to reduce the role of these inflammatory mechanisms. Furthermore, residual flexibility, hydrogen bonding, and structural packing were reported with uniform trajectories, showing no significant perturbation throughout the simulation. The protein motions within the simulation trajectories were clustered using principal component analysis (PCA). The IL-1ß-Inulin complex, approximately 70% of the total motion was attributed to the first three eigenvectors, while the remaining motion was contributed by the remaining eigenvectors. In contrast, for the COX2-Inulin complex, 75% of the total motion was attributed to the eigenvectors. Furthermore, in the iNOS-Inulin complex, the first three eigenvectors contributed to 60% of the total motion. Furthermore, the iNOS-Inulin complex contributed 60% to the total motion through the first three eigenvectors. To explore thermodynamically favorable changes upon mutation, motion mode analysis was carried out. The Free Energy Landscape (FEL) results demonstrated that the IL-1ß-Inulin achieved a single conformation with the lowest energy, while COX2-Inulin and iNOS-Inulin exhibited two lowest-energy conformations each. IL-1ß-Inulin and COX2-Inulin displayed total binding free energies of - 27.76 kcal/mol and - 37.78 kcal/mol, respectively, while iNOS-Inulin demonstrated the best binding free energy results at - 45.89 kcal/mol. This indicates a stronger pharmacological potential of iNOS than the other two complexes. Thus, further experiments are needed to use inulin to target iNOS and reduce DSS-induced colitis and other autoimmune diseases.

Long-term oral administration of burdock fructooligosaccharide alleviates DSS-induced colitis in mice by mediating anti-inflammatory effects and protection of intestinal barrier function.

BACKGROUND: Ulcerative colitis, a typical subtype of inflammatory bowel disease, can cause many serious complications. Burdock fructooligosaccharide (BFO), a linear inulin with a purity of 99.439% and a molecular weight of 2345 Da, demonstrates anti-inflammatory and immunomodulatory properties. METHODS: The Kunming mice were divided into two experimental models: a normal pretreatment model and a colitis experimental model. During the experimental treatment period, we assessed changes in weight and disease activity index (DAI), quantified the intestinal index, and determined myeloperoxidase (MPO) activity and reactive oxide species (ROS) levels in colitis mice. We also photographed colon morphology to investigate alterations in the integrity of the intestinal barrier function. Finally, we performed ELISA and qRT-PCR to evaluate the anti-inflammatory effect of BFO treatment on colitis mice. RESULT: The long-term oral administration of BFO alone exhibited protective effects by preventing disruption of the intestinal functional structure and increasing the colon index in mice. However, in a dextran sodium sulfate (DSS)-induced colitis mouse model, BFO administration facilitated quick recovery of body weight and effectively reduced the DAI, especially in the BFO-H group (500 mg/kg/day). BFO treatment maintained the integrity of the intestinal barrier by attenuating the crypt distortion and increasing the goblet cells count It restored the DSS-induced colon shortening and reduced the symptoms of colitis. These effects may be attributed to the appropriate concentrations of BFO effectively inhibiting MPO activity, clearing excessive ROS, and relieving spleen abnormalitie. BFO also attenuated the overexpression and excessive secretion of inflammatory cytokines (TNF-α, IL-1ß, IL-6, and MCP-1) induced by DSS, reduced intestinal inflammation, and consequently protected the intestinal barrier function. CONCLUSION: BFO effectively alleviated the symptoms of DSS-induced colitis by mediating anti-inflammatory effects and protecting the intestinal barrier integrity, thereby potentially facilitating the utilization of safer and more efficacious polysaccharides for managing chronic inflammatory diseases.

Immunomodulatory mechanisms of an acidic polysaccharide from the fermented burdock residue by Rhizopus nigricans in RAW264.7 cells and cyclophosphamide-induced immunosuppressive mice.

RBAPS is an acidic polysaccharide extracted from the burdock residue fermentation by Rhizopus nigricans. In RBAPS-activated RAW264.7 cells, transcriptome analysis identified a total of 1520 differentially expressed genes (DEGs), including 1223 down-regulated genes and 297 up-regulated genes. DEGs were enriched in the immune-related biological processes, involving in Mitogen-activated protein kinase (MAPK) and Toll-like receptor (TLR) signaling pathway, according to Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. The results of the confocal laser scanning microscope (CLSM) observation, antibody neutralization and Western blot verified that RBAPS modulated macrophages activation and cytokines secretion mainly via TLR4/MAPK/NF-κB signaling pathway. The immunomodulatory activity in vivo of RBAPS was investigated in cyclophosphamide (CTX)-induced immunosuppressive mice. RBAPS promoted the counts of white blood cells (WBC), red blood cells (RBC) and platelets (PLT) as well as the levels of immunoglobulins and cytokines (IgG, IgM, TNF-α, and IL-2) in immunosuppressive mice. RBAPS protected the spleen and thymus from CTX-induced injury by increasing the organ indexes, attenuating pathological damage, and promoting splenic lymphocytes proliferation. Importantly, RBAPS ameliorated the intestine integrity and function by promoting the expression of Occuldin, Claudin-5, Atg5, and Atg7, activating TLR4/MAPK signaling pathway in CTX-induced mice. This study suggested that RBAPS was a prime candidate of immunologic adjuvant in chemotherapy for the nutraceutical and pharmaceutical application.

Effects of burdock inulin-type fructans exposure on the physiological function of healthy mice and their filial generation.

Inulin-type fructans (ITFs) have been shown to possess various biological activities. However, studies on their safety and side effects are limited. Hence, we aimed to evaluate the possible effects of burdock ITFs on the physiological indices of healthy mice and their filial generation when fed for six months. Thirty-two C57BL/6J mice were randomly divided into two groups; a normal control (NC) and an ITFs group. The parental generations were kept in one cage with free access to a normal diet and double-distilled water (P-NC group) or burdock ITFs drinking water (P-ITFs group, 2% w/v). The filial generations (F-NC group and F-ITFs group) were kept separately and were fed as their parental generation. Behavior, organ/body weight, serum indices, histopathology, time of production, and number of pup births were observed. There were no significant adverse effects on these indices. Functional indices of the spleen, lung, heart, and pancreas of the ITFs groups were higher than those of the NC groups, respectively. Interestingly, the serum glucose (GLU), total cholesterol (TC), uric acid (UA) and creatine kinase (CK) levels of the ITFs groups were lower than those of the NC groups. Meanwhile, the pregnancy number and pup birth number of the P-ITFs group were more than those of P-NC group. Therefore, long-term consumption of burdock ITFs has no obvious adverse effects on the health of parental mice and their offspring, but may contribute to reproductive capacity, fatigue reduction, and risk reduction of renal disease.

Separation and Structural Characterization of a Novel Exopolysaccharide from Rhizopus nigricans.

The present study aims to analyze the structural characterization and antioxidant activity of a novel exopolysaccharide from Rhizopus nigricans (EPS2-1). For this purpose, EPS2-1 was purified through DEAE-52, Sephadex G-100, and Sephadex G-75 chromatography. The structural characterization of EPS2-1 was analyzed using high-performance gel permeation chromatography (HPGPC), Fourier transform infrared spectroscopy (FT-IR), methylation analysis, nuclear magnetic resonance (NMR) spectra, transmission electron microscope (TEM), and atomic force microscope (AFM). The results revealed that EPS2-1 is composed of mannose (Man), galactose (Gal), glucose (Glc), arabinose (Ara), and Fucose (Fuc), and possesses a molecular weight of 32.803 kDa. The backbone of EPS2-1 comprised →2)-α-D-Manp-(1→ and →3)-ß-D-Galp-(1→, linked with the O-6 position of (→2,6)-α-D-Manp-(1→) of the main chain is branch α-D-Manp-(1→6)-α-D-Manp-(1→, linked with the O-6 positions of (→3)-ß-D-Galp-(1→) of the main chain are branches →4)-ß-D-Glcp-(1→ and →3)-ß-D-Galp-(1→, respectively. Finally, we demonstrated that EPS2-1 also shows free radical scavenging activity and iron ion reducing ability. At the same time, EPS2-1 could inhibit the proliferation of MFC cells and increase the cell viability of RAW264.7 cells. Our results suggested that EPS2-1 is a novel polysaccharide, and EPS2-1 has antioxidant activity. In addition, EPS2-1 may possess potential immunomodulatory and antitumor activities. This study promoted the application of EPS2-1 as the functional ingredients in the pharmaceutical and food industries.

Separation, Purification, Structural Characterization, and Anticancer Activity of a Novel Exopolysaccharide from Mucor sp.

Mucor sp. has a wide range of applications in the food fermentation industry. In this study, a novel exopolysaccharide, labeled MSEPS, was separated from Mucor sp. fermentation broth through ethanol precipitation and was purified by ion-exchange chromatography, as well as gel filtration column chromatography. MSEPS was composed mostly of mannose, galactose, fucose, arabinose, and glucose with a molar ratio of 0.466:0.169:0.139:0.126:0.015 and had a molecular weight of 7.78 × 104 Da. The analysis of methylation and nuclear magnetic resonance results indicated that MSEPS mainly consisted of a backbone of →3,6)-α-d-Manp-(1→3,6)-ß-d-Galp-(1→, with substitution at O-3 of →6)-α-d-Manp-(1→ and →6)-ß-d-Galp-(1→ by terminal α-l-Araf residues. MTT assays showed that MSEPS was nontoxic in normal cells (HK-2 cells) and inhibited the proliferation of carcinoma cells (SGC-7901 cells). Additionally, morphological analysis and flow cytometry experiments indicated that MSEPS promoted SGC-7901 cell death via apoptosis. Therefore, MSEPS from Mucor sp. can be developed as a potential antitumor agent.

The flavonoid 3'-hydroxylase gene from the Antarctic moss Pohlia nutans is involved in regulating oxidative and salt stress tolerance.

Flavonoids are the important secondary metabolites. They are thought to play an important role in plant adaptation to terrestrial environment. However, the downstream branching pathway of flavonoids in bryophytes, which are the most ancient of terrestrial plants, remains unclear. Here, we cloned a flavonoid 3'-hydroxylase gene (PnF3'H) from the Antarctic moss Pohlia nutans and studied its function in plant stress tolerance. The Arabidopsis with overexpressing PnF3'H (AtOE) were constructed. The AtOE plants had more lateral roots and higher activities of antioxidant enzymes than the wild-type plants under oxidative stress. Meanwhile, the gene expression levels of reactive oxygen species (ROS) scavengers (i.e., AtCAT3, AtFeSOD1, and AtCu-ZnSOD3) were upregulated in the AtOE plants, and the transcription levels of ROS producing enzyme genes were significantly downregulated. The AtOE plans showed increased sensitivity to NaCl stress or abscisic acid (ABA) treatment during seed germination and early root development. Furthermore, several stress-resistant genes in the ABA signaling pathway were also downregulated in the AtOE plants when compared with the wild-type plants. These results suggested that PnF3'H participates in regulating the oxidative tolerance and ABA sensitivity to enable P. nutans to adapt to polar environments.

Effects of mixed culture fermentation of Bacillus amyloliquefaciens and Trichoderma longibrachiatum on its constituent strains and the biocontrol of tomato Fusarium wilt.

AIMS: To study the effects of mixed culture fermentation (MCF) of Bacillus amyloliquefaciens and Trichoderma longibrachiatum on its constituent strains and the application values for agricultural production, with the intention of developing efficient and environmentally friendly biocontrol agents. METHODS AND RESULTS: In this study, an in vitro antifungal growth experiment showed that the inhibitory rate of the MCF broth on pathogenic fungi (Fusarium oxysporum f. sp. lycopersici, Botrytis cinerea, Trichothecium roseum and Colletotrichum gloeosporioides) was less than that of B. amyloliquefaciens culture fermentation (BCF). Moreover, the content and gene expression of lipopeptide antibiotics were also lower than that in the BCF group. However, the pot experiments based on irrigation with appropriately diluted fermentation broth showed that the biocontrol effect of MCF on tomato Fusarium wilt was significantly higher than that of TCF (T. longibrachiatum culture fermentation) and BCF, and was approximately 15.79% higher than that of the BTF group which made by mixing equivalent amounts of BCF and TCF. In MCF broth, two micro-organisms antagonized and coexisted, and the growth of T. longibrachiatum was inhibited. Using transcriptomic analysis, we speculated that MCF can upregulate the expression of genes related to carbon and nitrogen metabolism, oxidation-reduction activity, sporulation, environmental information response and chemotaxis, and biosynthesis of secondary metabolites of B. amyloliquefaciens, which might enhance the nutrient substances metabolism and competitiveness, survival ability, colonization and adaptability to the environment to increase its biocontrol potential. CONCLUSIONS: Mixed culture fermentation could promote the more reasonable and effective utilization of biocontrol micro-organisms though improving biocontrol effect, enhancing strains survival and competitiveness, increasing beneficial metabolites, combined with resistance induction or synergistic control. SIGNIFICANCE AND IMPACT OF THE STUDY: Using MCF agronomically utilizes biocontrol agents in an efficient way, which has a good potential for commercial implementation and could reduce production costs.

Burdock Fructooligosaccharide Attenuates High Glucose-Induced Apoptosis and Oxidative Stress Injury in Renal Tubular Epithelial Cells.

Hyperglycemia-induced apoptosis and oxidative stress injury are thought to play important roles in the pathogenesis of diabetic nephropathy (DN). Attenuating high glucose (HG)-induced renal tubular epithelial cell injury has become a potential approach to ameliorate DN. In recent years, burdock fructooligosaccharide (BFO), a water-soluble inulin-type fructooligosaccharide extracted from burdock root, has been shown to have a wide range of pharmacological activities, including antiviral, anti-inflammatory, and hypolipidemic activities. However, the role and mechanism of BFO in rat renal tubular epithelial cells (NRK-52E cells) have rarely been investigated. The present study investigated the protective effect of BFO on HG-induced damage in NRK-52E cells. BFO could protect NRK-52E cells against the reduced cell viability and significantly increased apoptosis rate induced by HG. These anti-oxidative stress effects of BFO were related to the significant inhibition of the production of reactive oxygen species, stabilization of mitochondrial membrane potential, and increased antioxidant (superoxide dismutase and catalase) activities. Furthermore, BFO increased the expression of Nrf2, HO-1, and Bcl-2 and decreased the expression of Bax. In conclusion, these findings suggest that BFO protects NRK-52E cells against HG-induced damage by inhibiting apoptosis and oxidative stress through the Nrf2/HO-1 signaling pathway.

Preparation and characterization of sulfated inulin-type fructans from Jerusalem artichoke tubers and their antitumor activity.

The modification of polysaccharides is important for enhancing their biological activities. In this study, a pure inulin-type fructan, denoted as Jerusalem artichoke polysaccharide (P-JAP), was purified from Jerusalem artichoke tubers and modified by sulfation via treatment with a sulfur trioxide-pyridine complex to produce its sulfated derivative (S-JAP). Fourier-transform infrared spectroscopic analysis confirmed the successful introduction of sulfate groups. The inhibitory effects of S-JAP on the proliferation of human liver hepatocellular carcinoma (HepG2) cells was evaluated via a CCK-8 assay, and the pro-apoptotic effects were assessed using annexin V-FITC/PI double staining. The inhibition rates of various concentrations of S-JAP on HepG2 cells after 24, 48, and 72 h were significantly higher than those of P-JAP; moreover, S-JAP succeeded in promoting cell apoptosis. Thus, the sulfate-modified polysaccharide extracted from Jerusalem artichoke tubers was shown to exhibit effective antitumor activity with potential for further development.

Enhanced antitumor activity of inulin-capped Se nanoparticles synthesized using Jerusalem artichoke tubers.

An inulin polysaccharide with a molecular weight of ~ 2600 Da was derived from Jerusalem artichoke tubers and referred to as "JAP". Previous studies have shown that inulin can improve glucose tolerance and the liver lipid profile; however, its antitumor activity remains to be examined in detail. Therefore, to investigate the possible improvement of the antitumor activity of JAP, a novel nanostructured biomaterial was constructed by capping Se nanoparticles with JAP using sodium selenite, via a redox reaction with ascorbic acid, and referred to as "JAP-SeNPs". Transmission electron microscopy revealed that the average diameter of JAP-SeNPs is ~ 50 nm, and the C:Se mass ratio in JAP-SeNPs was found to be 15.4:1 by energy-dispersive X-ray spectroscopy. The well-dispersed JAP-SeNPs exhibited a significant in vitro antiproliferative effect on mouse forestomach carcinoma cells at a concentration of 400 µg/mL when incubated for 48 h, with an inhibition rate of 41.5%. Moreover, 38.9% of later apoptotic cells were observed. These results reveal that a combination of Se and JAP can effectively enhance the antitumor activity of polysaccharides obtained from Jerusalem artichoke tubers.

Structural and Biophysical Investigation of the Key Hotspots on the Surface of Epstein-Barr Nuclear Antigen 1 Essential for DNA Recognition and Pathogenesis.

Epstein-Barr Virus (EBV) is considered the most important human pathogen due to its role in infections and cellular malignancies. It has been reported that this Oncolytic virus infects 90% world's population. EBNA1 is required for DNA binding and survival of the virus and is considered an essential drug target. The biochemical and structural properties of this protein are known, but it is still unclear which residues impart a critical role in the recognition of dsDNA. Intending to disclose only the essential residues in recognition of dsDNA, this study used a computational pipeline to generate an alanine mutant of each interacting residue and determine the impact on the binding. Our analysis revealed that R469A, K514A, Y518A, R521A and R522A are the key hotspots for the recognition of dsDNA by the EBNA1. The dynamics properties, i.e. stability, flexibility, structural compactness, hydrogen bonding frequency, binding affinity, are altered by disrupting the protein-DNA contacts, thereby decreases the binding affinity. In particular, the two arginine substitution, R521A and R522A, significantly affected the total binding energy. Thus, we hypothesize that these residues impart a critical role in the dsDNA recognition and pathogenesis. This study would help to design structure-based drugs against the EBV infections.

The moss flavone synthase I positively regulates the tolerance of plants to drought stress and UV-B radiation.

Flavonoids are extensively distributed secondary metabolites in land plants. They play a critical role in plant evolution from aquatic to terrestrial and plant adaption to ultraviolet radiation. However, the downstream branching pathway of flavonoids and its regulatory mechanism in bryophytes, which are the most ancient of terrestrial plants, remain unclear. Here, a type I flavone synthase (PnFNSI) was characterized from the Antarctic moss Pohlia nutans. PnFNSI was primarily distributed in the cytoplasm, as detected by subcellular localization. PnFNSI could catalyze the conversion of naringenin to apigenin with an optimal temperature between 15 and 20 °C in vitro. Overexpression of PnFNSI in Arabidopsis alleviated the growth restriction caused by naringenin and accumulated apigenin product. PnFNSI-overexpressing plants showed enhanced plant tolerance to drought stress and UV-B radiation. PnFNSI also increased the enzyme activities and gene transcription levels of reactive oxygen species (ROS) scavengers, protecting plants against oxidative stress. Moreover, overexpression of PnFNSI enhanced the flavone biosynthesis pathway in Arabidopsis. Therefore, this moss FNSI-type enzyme participates in flavone metabolism, conferring protection against drought stress and UV-B radiation.

Signaling pathways associated with macrophage-activating polysaccharide isolated from the fermentation liquor of Rhizopus nigricans.

Our previous reports showed that the structural features and immunologic enhancement of polysaccharide (EPS1-1) from Rhizopus nigricans. However, the molecular mechanism in cellular immunomodulatory of EPS1-1 remains unclear. Here the experiments for the molecular mechanisms of EPS1-1 on the peritoneal macrophages were performed. The results demonstrated that the expression of TLR4 was significantly improved by EPS1-1. Subsequently, the phosphorylation of p38MAPK, ERK1/2, JNK and IKKα/ß were promoted. Moreover, EPS1-1 enhanced the expressions of IL-2, TNF-α and iNOS in EPS1-1-induced macrophages which were pretreated with MAPK signaling pathway inhibitors, and reduced the blocking effects of the inhibitors to the expressions of p-p38MAPK, p-ERK1/2 and p-IKKα/ß. Therefore, these results illustrated that EPS1-1 could improve the immune functions of peritoneal macrophages by promoting the gene expressions of IL-2, TNF-α and iNOS via the MAPK and NF-κB signaling pathways.

Exopolysaccharides isolated from Rhizopus nigricans induced colon cancer cell apoptosis in vitro and in vivo via activating the AMPK pathway.

Colorectal cancer (CRC) is a leading cause of cancer-related human deaths. The exopolysaccharide (EPS1-1), isolated from Rhizopus nigricans, has been described as exhibiting anti-tumor and pro-apoptotic activity against CRC, although the underlying mechanism is poorly understood. Herein, we investigate how EPS1-1 induces apoptosis of CRC cells in vitro and in vivo. Our results show that, in vitro, EPS1-1 suppressed cell growth and facilitated apoptosis in a dose- and time-dependent manner by activating the AMP-activated protein kinase (AMPK) pathway in mouse colon cancer CT26 cells. However, treatment with small interfering RNAs (siRNAs) targeting AMPKα or with compound C, an AMPK inhibitor, interfered with the pro-apoptosis effects of EPS1-1. We also show that EPS1-1 initiated the release of reactive oxygen species (ROS) and liver kinase B1 (LKB1), both of which are necessary signals for AMPK activation. Furthermore, EPS1-1-mediated apoptosis is regulated by inactivation of mammalian target of rapamycin complex 1 (mTORC1) and activation of the jun-NH2 kinase (JNK)-p53 signaling axis dependent on AMPK activation. In vivo, azoxymethane/dextran sulfate sodium (AOM/DSS)-treated CRC mice, when administered EPS1-1, exhibited activation of the AMPK pathway, inhibition of mTORC1, and accumulation of p53 in tumor tissues. Collectively, these findings suggest that EPS1-1-induced apoptosis relies on the activation of the AMPK pathway. The present study provides evidence suggesting that EPS1-1 may be an effective target for development of novel CRC therapeutic agents.

Metabonomic Variation of Exopolysaccharide from Rhizopus nigricans on AOM/DSS-Induced Colorectal Cancer in Mice.

BACKGROUND: Colorectal cancer (CRC), which occurs at the junction of the rectum and sigmoid colon, is a common malignancy associated with poor prognosis and high mortality worldwide. The exopolysaccharide (EPS1-1), isolated from the fermentation broth of Rhizopus nigricans (R. nigricans), has been reported to possess anti-CRC properties. However, the metabolic alterations caused by azoxymethane (AOM) and dextran sulfate sodium (DSS) are still unknown. METHODS: In the present study, a mice colon cancer model was established by treatment with AOM/DSS. LC-MS/MS-based metabolomics studies were performed to analyze metabolic alterations at the tissue level. Partial least squares discriminant analysis (PLS-DA) was used to identify differentially expressed metabolites. RESULTS: Nineteen distinct metabolites were identified that were associated with disruptions in the following pathways: biosynthesis of unsaturated fatty acids, pyrimidine metabolism, phenylalanine metabolism, fatty acid metabolism, folate biosynthesis, and inositol phosphate metabolism. Furthermore, six significantly altered metabolites were involved in these six pathways. Compared with the Model group, the expression of cytosine, deoxyuridine, 20-hydroxy-leukotriene E4, and L-homocysteic acid was lower, whereas that of 2-dehydro-3-deoxy-6-phospho-D-gluconic acid and hematoporphyrin was higher in the EPS1-1 group. CONCLUSION: The results of multivariate statistical analysis demonstrate a promising application of the above metabolites by EPS1-1 in CRC therapy. Deeper understanding of the related mechanism warrants further investigation.

Carbon Nanoparticles Inhibit Α-Glucosidase Activity and Induce a Hypoglycemic Effect in Diabetic Mice.

New, improved therapies to reduce blood glucose are required for treating diabetes mellitus (DM). Here, we investigated the use of a new nanomaterial candidate for DM treatment, carbon nanoparticles (CNPs). CNPs were prepared by carbonization using a polysaccharide from Arctium lappa L. root as the carbon source. The chemical structure and morphology of the CNPs were characterized using Fourier-transform infrared spectroscopy, X-ray photoelectron spectroscopy, elemental analysis, and transmission electron microscopy. CNPs were spherical, 10-20 nm in size, consisting of C, H, O, and N, and featuring various functional groups, including C=O, C=C, C-O, and C-N. In vitro, the as-prepared CNPs could inhibit α-glucosidase with an IC50 value of 0.5677 mg/mL, which is close to that of the reference drug acarbose. Moreover, in vivo hypoglycemic assays revealed that the CNPs significantly reduced fasting blood-glucose levels in mice with diabetes induced by high-fat diet and streptozocin, lowering blood glucose after intragastric administration for 42 days. To the best of our knowledge, this is the first report of CNPs exhibiting α-glucosidase inhibition and a hypoglycemic effect in diabetic mice. These findings suggest the therapeutic potential of CNPs for diabetes.

PnSAG1, an E3 ubiquitin ligase of the Antarctic moss Pohlia nutans, enhanced sensitivity to salt stress and ABA.

Plant U-box (PUB) E3 ubiquitin ligases play crucial roles in the plant response to abiotic stress and the phytohormone abscisic acid (ABA) signaling, but little is known about them in bryophytes. Here, a representative U-box armadillo repeat (PUB-ARM) ubiquitin E3 ligase from Antarctic moss Pohlia nutans (PnSAG1), was explored for its role in abiotic stress response in Arabidopsis thaliana and Physcomitrella patens. The expression of PnSAG1 was rapidly induced by exogenous abscisic acid (ABA), salt, cold and drought stresses. PnSAG1 was localized to the cytoplasm and showed E3 ubiquitin ligase activity by in vitro ubiquitination assay. The PnSAG1-overexpressing Arabidopsis enhanced the sensitivity with respect to ABA and salt stress during seed germination and early root growth. Similarly, heterogeneous overexpression of PnSAG1 in P. patens was more sensitive to the salinity and ABA in their gametophyte growth. The analysis by RT-qPCR revealed that the expression of salt stress/ABA-related genes were downregulated in PnSAG1-overexpressing plants after salt treatment. Taken together, our results indicated that PnSAG1 plays a negative role in plant response to ABA and salt stress.

Structure and Anti-Tumor Activities of Exopolysaccharides from Alternaria mali Roberts.

In this study, an extracellular polysaccharide from Alternaria mali Roberts (AMEP) was extracted, and its structure was characterized, in addition to its antitumor activity in vitro. Neutral polysaccharide AMEP-1 and anionic polysaccharide AMEP-2 were isolated from AMEP, and their monosaccharide compositions consisted of mannose (Man), glucose (Glc), and galactose (Gal) but at different ratios. The linking mode of both AMEP-1 and AMEP-2 is Manp-(1→4) and Glcp-(1→6), and the branched chains are connected to the main chain through O-6. AMEP-2 inhibited the proliferation of BGC-823 cells in a time- and concentration-dependent manner. AMEP-2 also induced the apoptosis of BGC-823 cells, and showed anti-tumor effects by inducing cell cycle arrest in the S phase, reactive oxygen species production, and mitochondrial membrane potential reduction in BGC-823 cells. Therefore, AMEP-2 shows potential for further development as a novel anti-tumor agent.

The inhibitory effect of polysaccharide from Rhizopus nigricans on colitis-associated colorectal cancer.

An extracellular polysaccharide (EPS1-1) of Rhizopus nigricans was found to enhance immunity and reduce colon cancer cell proliferation. Here, the effect of EPS1-1 on a mouse model of colitis-associated cancer (CAC) induced by azoxymethane (AOM)/dextran sodium sulfate (DSS) was investigated. Pathological symptoms, including weight loss, piloerection, hematochezia and insensitivity caused by AOM/DSS, were relieved by EPS1-1. Anatomical results showed a 100% tumor incidence, a series of neoplasms, disordered cell structure and hyperplastic glands in the model group, while the abnormal behaviors were relieved and the tumors decreased in the EPS1-1 group. Compared with the model group, the EPS1-1 group showed decreased oncogenic protein (COX-2, ß-catenin, CyclinD1 and C-Myc) expression. TUNEL staining showed that EPS1-1 increased the apoptosis of colon cancer cells in mice. Furthermore, the expression of proliferative proteins (Ki-67 and PCNA) and an antiapoptotic gene transcript (Bcl-2) were significantly down regulated by EPS1-1, while apoptotic gene transcripts (p53 and Bax) were enhanced. In addition, EPS1-1 notably decreased the number of cells positive for CD68, F4/80 and NF-κB and reduced the concentrations of inflammatory factors (TNF-α and IL-6) in serum compared with those in the model group. Taken together, these results suggest that EPS1-1 may be a therapeutic option for the prevention and treatment of CAC.