Development of Glycosylation-Modified DPPA-1 Compounds as Innovative PD-1/PD-L1 Blockers: Design, Synthesis, and Biological Evaluation.

In the context of peptide drug development, glycosylation plays a pivotal role. Accordingly, L-type peptides were synthesized predicated upon the PD-1/PD-L1 blocker DPPA-1. Subsequent glycosylation resulted in the production of two distinct glycopeptides, D-glu-LPPA-1 and D-gal-LPPA-1, by using D-glucose (D-glu) and D-galactose (D-gal), respectively, during glycosylation. Both glycopeptides significantly inhibited the interaction between PD-1 and PD-L1, and the measured half maximal inhibitory concentrations (IC50s) were 75.5 µM and 101.9 µM for D-glu-LPPA-1 and D-gal-LPPA-1, respectively. Furthermore, D-gal-LPPA-1 displayed a pronounced ability to restore T-cell functionality. In an MC38 tumor-bearing mouse model, D-gal-LPPA-1 demonstrated a significant inhibitory effect. Notably, D-gal-LPPA-1 substantially augmented the abundance and functionality of CD8+ T cells in the tumor microenvironment. Additionally, in the lymph nodes and spleens, D-gal-LPPA-1 significantly increased the proportion of CD8+ T cells secreting interferon-gamma (IFN-γ). These strong findings position D-gal-LPPA-1 as a potent enhancer of the antitumor immune response in MC38 tumor-bearing mice, underscoring its potential as a formidable PD-1/PD-L1 blocking agent.

Trichoderma hamatum and Its Benefits.

Trichoderma hamatum (Bonord.) Bainier (T. hamatum) belongs to Hypocreaceae family, Trichoderma genus. Trichoderma spp. are prominently known for their biocontrol activities and plant growth promotion. Hence, T. hamatum also possess several beneficial activities, such as antimicrobial activity, antioxidant activity, insecticidal activity, herbicidal activity, and plant growth promotion; in addition, it holds several other beneficial properties, such as resistance to dichlorodiphenyltrichloroethane (DDT) and degradation of DDT by certain enzymes and production of certain polysaccharide-degrading enzymes. Hence, the current review discusses the beneficial properties of T. hamatum and describes the gaps that need to be further considered in future studies, such as T. hamatum's potentiality against human pathogens and, in contrast, its role as an opportunistic human pathogen. Moreover, there is a need for substantial study on its antiviral and antioxidant activities.

Antimicrobial activity and enzymatic analysis of endophytes isolated from Codonopsis pilosula.

The roots of the medicinal plant Codonopsis pilosula (Franch.) Nannf (C. pilosula) possess most medicinal supplements. In current research on C. pilosula root endophytes were isolated, identified, and evaluated for their antimicrobial activity against human pathogens such as Escherichia coli, Staphylococcus aureus, Bacillus subtilis, Salmonella typhi, and Pseudomonas aeruginosa and the fungi Candida albicans and Aspergillus niger. Endophytes C.P-8 and C.P-20 exhibited very significant antimicrobial activity, the secondary metabolite of C.P-8 registered at retention time 24.075 by HPLC analysis. Significant minimum inhibitory concentration (MIC) of C.P-8 was exhibited at 250 µg/ml against S. aureus and 500 µg/ml against B. subtilis. Qualitative, quantitative analyses, and partial purification of enzymes and purity was analysed by molecular weight determined by SDS‒PAGE of enzymes produced by C.P-20, amylase-64 kDa, protease-64 kDa, chitinase-30 kDa, and cellulase-54 kDa. Optimum pH and temperature of the partially purified enzymes, was carried out. The partially purified enzymes from C.P-20 displayed maximum activity at pH 6-7 and temperatures of 40°C-45°C. Moreover, the above endophytes will be useful tools for producing active enzymes and active bioantimicrobial agents against human pathogens.

Preparation and structural characterization of acid-extracted polysaccharide from Grifola frondosa and antitumor activity on S180 tumor-bearing mice.

In this study, an acid-extracted polysaccharide (GFP-A) was extracted from the fruiting bodies of G. frondosa with 1 % hydrochloric acid solution. Our study aimed to imitate the processes of digestion, absorption and antitumor activities of polysaccharides from G. frondosa under the acid environment of stomach in the body. The preliminary structural analysis resulted that GFP-A (about 1.10 × 106 Da) was a neutral polysaccharide composed of xylose, mannose, glucose (molar ratio: 0.12:1.00:6.98) with α-type glycosidic linkages. Additionally, antitumor activities on S180 tumor-bearing mice showed that GFP-A could effectively inhibit the growth of S180 tumor cells by protecting immune organs (thymus and spleen), activating immune cells (NK cells, lymphocytes and macrophages), upregulating the secretion of serum cytokines (TNF-α, IL-2 and IFN-γ) in vivo. H & E staining and cell cycle determination further demonstrated that GFP-A could induce S180 tumor cells apoptosis via arresting them in G1 phase. These results demonstrated that GFP-A could provide a theoretical basis for treatment of cancer.

Characterization of a polysaccharide from Polygala tenuifolia willd. with immune activity via activation MAPKs pathway.

Polysaccharides from the Polygala tenuifolia Willd. have been shown multiple biological activities, however the structural feature and immunomodulatory activity are still rarely reported. In this study, a polysaccharide was obtained by purification, and its structural characteristics and immune activity were analyzed. The polysaccharide was a homogeneous macromolecular polysaccharide with smooth flat flakes surface structure and molecular weight of 2.34 × 105 Da, and composed of Rha, Ara, Xyl, Man, Glc, Gal. Methylation and NMR analyses confirmed that the repeating unit of polysaccharide was [→3)-α-Araf-(1 â†’ 3)-α-Araf-(1 â†’ 5)-α-Araf-(1 â†’ 5)-α-Araf-(1 â†’ 3)-α-Araf-(1 â†’ ]n, and the side chain was α-Araf-(1 â†’ 6)-ß-Galp-(1 â†’ 6)-ß-Glcp-(1 â†’ 6)-α-Manp-(1→, which was attached to the C3 of â†’ 3,5)-α-Araf-(1 â†’. In vitro, the RAW 264.7 cells were co-cultivated with LPS and polysaccharide, and the results revealed that the polysaccharide can promote cell proliferation, activate effectors to release cytokines (TNF-α, IL-6, IL-1ß), and then activate macrophages for immune activity. Therefore, we can infer that the polysaccharide might regard as a potential immunomodulator.

Research trends in hypertension associated with obstructive sleep apnea: a bibliometric analysis.

OBJECTIVES: The purpose of this study was to systematically analyze the studies of hypertension associated with obstructive sleep apnea to assess the current status and hot spots in this field. METHODS: We searched the Web of Science Core Collection for publications related to hypertension associated with obstructive sleep apnea published before July 3, 2021. Bibliometric analyses and science mappings were carried out using the CiteSpace 5.8.R1 and Microsoft Office Excel 2019. CiteSpace 5.8.R1 was used to visualize the distribution of research fields, analyze co-occurring keywords and burst terms to detect trends and frontiers, and identify leading collaborations among countries, authors, and institutions. Microsoft Office Excel 2019 was used to make bar graphs, histograms and line graphs. RESULTS: According to the search strategy, a total of 7263 published articles and reviews were retrieved. The research on hypertension associated with obstructive sleep apnea has been developing quickly at present. Sleep and Breathing was the most productive journal. The USA was a major producing country and Harvard Medical School was the most productive institution in this field. In the field of hypertension associated with obstructive sleep apnea, the main research hotspots were continuous positive airway pressure, cardiovascular disease, and obesity. CONCLUSIONS: The present study provides a new perspective for the study of hypertension associated with obstructive sleep apnea and valuable information for researchers to find potential partners and cooperative institutions, hot issues and research frontiers.

Decoding the enhanced antioxidant activities of the combined small berry pomaces by widely targeted metabolomics analysis.

Small berry pomaces (SBPs) are poorly utilized as an inexpensive source of bioactive compounds. This study investigated the impact of compounding treatment on nutritional and antioxidant characteristics of combined SBPs, in comparison with single SBP. The results showed that the amounts of protein, minerals, dietary fiber (DF) and anthocyanidins were significantly (p < 0.05) higher in combined SBPs than in combined fruits. Moreover, the combined SBPs were characterized by an elevated abundance of minerals and anthocyanidins (6 kinds, and 5 kinds, respectively), substantiating the effectiveness of compounding treatment on SBP nutrition. A total of 776 secondary phytochemicals were detected in combined SBPs by a widely targeted metabolomics approach. Each SBP contained approximately 100 kinds of unique natural antioxidants. Furthermore, the combined SBPs group had the highest antioxidant activity compared with single SBP. Meanwhile, the antioxidant activities determined in combined SBPs were higher than arithmetic mean value of single SBP. The synergism and interaction of active components in different sources of SBPs play vital role in the high antioxidant capacity of combined SBPs. All the results provide reference for the comprehensive development and utilization of fruit residues. The SBPs should be highly prized for their substantial amount of nutritional and bioactive constituents, including protein, DF, essential minerals and secondary metabolites. These secondary metabolites are positively associated with antioxidant benefits. The present study summarizes the knowledge about bioactive compounds and antioxidant activities of combined SBPs group and discusses the relevant mechanisms. A conclusion can be educed that combined process is an effective way to improve properties of the pomaces.

Ultrasonic-Assisted Extraction of Codonopsis pilosula Glucofructan: Optimization, Structure, and Immunoregulatory Activity.

In recent years, multiple edible polysaccharides from Codonopsis pilosula were mainly isolated with high average molecular weights and exhibited various bioactivities, but it was proven that low-molecular-weight polysaccharides could exert stronger activities due to the superior water solubility and permeability. In the present study, the water-soluble polysaccharide C. pilosula with low molecular weight was isolated under ultrasonic assistance at 30 °C, the extraction process was optimized via response surface method (RSM), and the structure and immunoregulatory activity were further investigated. The maximum yield (4.86%) for crude polysaccharides (cCPPs) was obtained under following parameters: ultrasonic power of 370 W, liquid/material ratio of 33 mL/g, ultrasonic time of 81 min. Subsequently, the cCPPs were further purified through dialysis and Sephadex G-25 column to acquire purified polysaccharide (CPPs). Structural analysis indicated that CPPs was a glucofructan (average molecular weight of 4.23 × 103 Da) with (2→1)-ß-D-Fruf and (1→)-α-D-Glcp as the backbone branched by (2→6)-ß-D-Fruf. Additionally, CPPs could enhance immunoregulatory function by stimulating NO production and cytokine (IL-6 and TNF-α) secretion of RAW264.7 macrophages dose-dependently, which presented no cytotoxic effects. These data suggest that CPPs have the potential to be used as a nutritional dietary compound and natural immunostimulant supplement in the food industry.

Antitumor activity and immunomodulation mechanism of a novel polysaccharide extracted from Polygala tenuifolia Willd. evaluated by S180 cells and S180 tumor-bearing mice.

We recently isolated a polysaccharide from Polygala tenuifolia Willd. (PTP) and reported that such a PTP could induce cell apoptosis with FAS/FAS-L-mediated death receptor pathway in human lung cancer cells. Herein, we indicate antitumor activity and immunoregulation of PTP for S180 sarcoma cells by in vitro and in vivo targeting. In vitro, S180 cells took on prominent characteristics of apoptosis under-treated with PTP in follow-up antitumor activity studies, including irregular shrinkage and fragmentation nuclear, apoptotic bodies formation, and reduction of mitochondrial membrane potential (MMP). Additionally, flow cytometry indicated that the number of normal cells (FITC-/PI-) gradually decreased from 98.08% to 16.31%, while the number of apoptotic cells (FITC+/PI- or FITC+/PI+) increased from 0.87% to 54.84%. The ratio of BAX and Bcl-2 increased, which promoted the release of Cytochrome C (CytC), and it further maximized the expression of activated-caspase-9/-3. Additionally, the PTP revised the immune organ indexes, the activities of NK cells and lymphocytes, and induced the secretion of IL-2 (7.34-16.17%), IFN-γ (14.34-20.85%) and TNF-α (12.32-22.58%) in vivo. Thus, PTP can induce cell apoptosis and activate the immunoregulation mechanism thereby exhibiting biological activity.

Structural characterization of a water-soluble polysaccharide from Angelica dahurica and its antitumor activity in H22 tumor-bearing mice.

A novel Angelica dahurica polysaccharide (ADP) with Mw of 6.09 × 103 Da was isolated. The contents of total sugar and uronic acid in ADP were 91.04% and 12.69%. The structure characteristics indicated that ADP was an acidic polysaccharide consisting of rhamnose, arabinose, galactose, glucose, mannose, glucuronic acid and galacturonic acid (0.09: 0.61: 1.88: 1: 0.14: 0.63: 0.03). Moreover, there were →3)-Manp-(1→, →4, 6)-Galp-(1→, →4)-Galp-(1→, →3)-Glcp-(1→, →5)-Araf-(1→, →2)-Galp-(1→ in ADP with relative molar ratios of 0.32:0.57:0.29:0.95:0.71:0.26. In vivo experiments suggested that ADP significantly inhibited the tumor growth of mice, increased the activities of spleen lymphocytes and natural killer (NK) cells, improved the cytokine level (IL-2 and TNF-α) and the proportions of lymphocyte subsets in the peripheral blood. The tumor cell progression was arrested in the G1 phase, and the apoptosis rate of tumor cells were 7.54% and 19.32% at the dose of 100 and 200 mg/kg, which was consistent with the results of pathological observation. In summary, the study might provide a theoretical basis for the application on functional foods containing Angelica dahurica polysaccharides.

Antitumor and immunoregulatory activities of a novel polysaccharide from Astragalus membranaceus on S180 tumor-bearing mice.

The Astragalus membranaceus polysaccharide (APS4) with direct cytotoxicity on various cancer cells has been prepared in our previous study, while the underlying therapeutic role of APS4 on solid tumors in vivo hasn't been investigated yet. Therefore, in this paper, the lymphocytes-mediated antitumor and immunoregulatory activities of APS4 were researched by establishing S180 tumor-bearing mice model. Flow cytometry analysis revealed that APS4 could effectively regulate the percentages of CD3+, CD4+, CD8+ T cells and CD19+ B cells in thymus, peripheral blood and spleen of S180 tumor-bearing mice, dose-dependently. H&E staining and cell cycle determination of solid tumors manifested that APS4 treatment could significantly inhibit the growth of solid tumors by inducing cells apoptosis. Furthermore, two-dimensional electrophoresis and western blot analysis further demonstrated that APS4 could activate antitumor-related immune cells and promote anaerobic metabolism of tumor microenvironment, thereby causing the apoptosis of S180 tumor cells. These data implicated that APS4 could be used as a potential dietary supplement for immune enhancement.

Structural characteristics and anti-tumor/-oxidant activity in vitro of an acidic polysaccharide from Gynostemma pentaphyllum.

In this paper, a novel acidic polysaccharide (CPS-1) was successively prepared from Gynostemma pentaphyllum using hot water isolation method to explore its antitumor and antioxidant activities. Structural characteristics of CPS-1 were evaluated by SEM, HPGPC, HPAEC-PAD, FT-IR, and NMR. The results indicated: CPS-1 was mainly composed of Ara, Gal, Glc, Xyl, Man, GalA and GlcA in a molar ratio of 1.23:2.14:0.67:0.2:0.29:0.16:0.04 with molecular weight of 3297 kDa. Combining with the results of FT-IR and NMR, it was inferred that CPS-1 was mainly possessed the five main linkages including α-D-Ara, α-D-Gal, α-D-Man, α-D-Xyl and ß-D-Glc. Furthermore, MTT results exhibited that the IC50 value of CPS-1 for inhibitive effect on SPC-A-1 and MGC-803 cells for 24 h were 284.36 and 365.27 µg/mL, respectively. Microscopic observations showed that the cells exhibited significant apoptotic characteristics, such as cell shrinkage, the decreased of cell adherence and the appearance of apoptotic bodies. It was shown that CPS-1 had significant anti-tumor activity. In addition, the ability of CPS-1 to scavenge superoxide radical, ABTS and DPPH radicals was also enhanced with the increased of concentration. Therefore, it was revealed that CPS-1 might be used as a natural anticancer and antioxidant component.

A novel polysaccharide from Castanea mollissima Blume: Preparation, characteristics and antitumor activities in vitro and in vivo.

A new water-soluble polysaccharide, CMP90, with a molecular weight of 23.9 kDa was isolated from Castanea mollissima Blume and the preliminary structural characteristics and antitumor effects of CMP90 in vitro and in vivo were investigated in the research. CMP90 consists of arabinose, galactose, glucose, xylose and mannose (molar ratio: 0.08:0.11:5.14:0.12:0.08) with α- and ß-anomeric units. The results of in vitro experiments indicated that CMP90 exhibited a significant inhibitory effect on the proliferation of HL-60 cells with typical apoptotic characteristics by inducing cell cycle arrested at G1/M phase. Additionally, the results in vivo suggested CMP90 was able to inhibit the growth of S180 solid tumors via protecting immune organs, improving the levels of serum cytokines (TNF-α, IL-2 and IFN-γ), enhancing the activities of immune cells (macrophages, lymphocytes and NK cells) and inducing cell apoptosis or death. Taken together, these combined data clearly indicated that CMP90 may be used as a potential candidate agent for cancer therapy.

FAS/FAS-L-mediated apoptosis and autophagy of SPC-A-1 cells induced by water-soluble polysaccharide from Polygala tenuifolia.

In this study, a water-soluble polysaccharide was extracted from Polygonum tenuifolia to inhibit the proliferation of SPC-A-1 cells by the FAS/FAS-L-mediated pathway and autophagy. The molecular weight, composition, and structure of the Polygala tenuifolia polysaccharide (PTP) was detected by HPLC, HPAEC-PAD, NMR and FT-IR. The purified polysaccharide was composed of Ara, Gal, and Glc (molar ratio: 2.6:1.8:1.0) with α- and ß-configurations. Morphological changes were observed with microscopes, and the cell apoptosis-related markers detected by flow cytometry indicated that apoptosis and autophagy occurred in the SPC-A-1 cells. Western blot analysis showed that the expression of proteins was related to apoptosis and autophagy. The death receptor pathway demonstrated the up-regulated expression of FAS, ligand FAS-L, and FADD, which led to a cascade reaction of the caspase family that induced cell apoptosis. The up-regulation of LC 3B-II and the down-regulation of P62 indicated the occurrence of autophagy. In summary, these results showed that PTP can induce FAS/FAS-L-mediated apoptosis and autophagy in SPC-A-1 cells, and provide a strong theoretical basis for tumor prevention and clinical application of PTP in the future.

An alcohol-soluble polysaccharide from Atractylodes macrocephala Koidz induces apoptosis of Eca-109 cells.

In this study, polysaccharides from Atractylodes macrocephala Koidz (APA) which were soluble in alcohol were prepared, purified, analyzed the structure and investigated the antitumor activity in vitro cell experiment. Results of high-performance gel permeation chromatography (HPGPC), fourier-transform infrared spectroscopy (FT-IR), and gas chromatography (GC) showed that APA was a 2.1KDa neutral hetero polysaccharide composed of arabinose and glucose (molar ratio, 1.00:4.57) with pyranose rings and α-type and ß-type glycosidic linkages. Results by MTT experiments showed that the proliferation inhibition was 74.63% in Eca109 cells treated with 2 mg/mL dose of APA. Annexin V/PI assay, Hoechst 33,258 staining, cell cycle distribution, rhodamine 123 dye assay and western blot assay clarified that APA could accelerate the apoptosis of Eca109 cells by mitochondrial pathway and stocked cells at S phase. These data indicated that APA is a promising potential candidate for therapeutic treatment of esophageal cancer.

Antitumor effects of seleno-short-chain chitosan (SSCC) against human gastric cancer BGC-823 cells.

Seleno-short-chain chitosan (SSCC) is a derivative of chitosan. In the present study, we sought to investigate the underlying antitumor mechanism of SSCC on human gastric cancer BGC-823 cells in vitro. MTT assay suggested that SSCC exhibited a dose-dependent inhibitory effect on the proliferation of BGC-823 cells. We found the SSCC-treated cells showed typical morphological characteristics of apoptosis in a dose dependent manner by observing on microscope. Annexin V-FITC/PI double staining and cell cycle assay identified that SSCC could induce BGC-823 cells apoptosis by triggering G2/M phase arrest. Our research provided the first evidence that SSCC could effectively induce the apoptosis of BGC-823 cells via an intrinsic mitochondrial pathway, as indicated by inducing the disruption of mitochondrial membrane potential (MMP), the excessive accumulation of reactive oxidative species (ROS), the increase of Bax/Bcl-2 ratio and the activation of caspase 3, caspase 9 and cytochrome C (Cyt-C) in BGC-823 cells. These combined results clearly indicated that SSCC could induce BGC-823 cells apoptosis by the involvement of mitochondrial signaling pathway, which provided precise experimental evidence for SSCC as a potential agent in the prevention and treatment of human gastric cancer.

Polysaccharide extracted from Atractylodes macrocephala Koidz (PAMK) induce apoptosis in transplanted H22 cells in mice.

Polysaccharide of Atractylodes macrocephala Koidz (PAMK) was extracted by alcohol sedimenting ranging from 60% to 90% and purified by ultrafiltration membrane. The high-performance gel permeation chromatography (HPGPC), Fourier-transform infrared spectroscopy (FT-IR), gas chromatography (GC) and Nuclear magnetic resonance (NMR) revealed that PAMK was a 4.1KDa neutral heteropolysaccharide composed of galactose, arabinose and glucose with α-configuration (molar ratio, 1: 1.5: 5). Results of determination of chemical components suggested that PAMK contained 96.47% of polysaccharide and little protein, nucleic acid and uronic acid. Antitumor experiments in vivo could be concluded that PAMK had a significantly cytotoxic and anti-tumor effect by blocking tumor cells in S phase. And not only that, PAMK could protect immune organ efficiently, comparing with cyclophosphamide. The research provided a potential antineoplastic drug component for tumor treatment.

Apoptosis of human gastric carcinoma MGC-803 cells induced by a novel Astragalus membranaceus polysaccharide via intrinsic mitochondrial pathways.

In our previous study, a novel cold-water-soluble polysaccharide (APS4) was isolated from Astragalus membranaceus. This study aimed to evaluate the proliferation inhibition and apoptosis-induced effects of APS4 on human gastric carcinoma MGC-803 cells and to investigate its potential molecular mechanism. It was found that APS4 could significantly suppress the proliferation of MGC-803 cells in a concentration- and time-dependent manner. Morphologic observations and Annexin V-FITC/PI staining showed that APS4-treated MGC-803 cells exhibited typical morphological characteristics of apoptosis. Cell cycle detection revealed that APS4 could arrest MGC-803 cells in S phase of the cell cycle. Additionally, APS4 treatment could induce the mitochondria-dependent apoptosis, which was closely related to the accumulation of intracellular ROS, the collapse of mitochondrial membrane potential, the increase of the pro-apoptotic/anti-apoptotic (Bax/Bcl-2) ratios, the release of cytochrome c, further activating the expression of caspase-9/-3 and the cleavage of poly-ADP-ribose polymerase (PARP) in MGC-803 cells. Taken together, our results suggested that APS4 had observable apoptosis-induced effects on MGC-803 cells via arresting the cell cycle in S phase and inducing the intrinsic mitochondrial apoptosis pathway.

Injectable Silk-Vaterite Composite Hydrogels with Tunable Sustained Drug Release Capacity.

Improving the efficiency of chemotherapy remains a key challenge in drug delivery. Many drug carriers have been designed to achieve multifunctional factors as part of their performance, including controlled release, dispersibility in aqueous environments, and targeting to cancer sites. However, it is difficult to optimize multiple properties simultaneously for a single carrier system. Here, synergistic carriers composed of vaterite microspheres and silk nanofiber hydrogels were developed to improve the dispersibility of vaterite spheres and the control of drug delivery without compromising the injectability or sensitivity to pH. The vaterite microspheres were dispersed homogeneously and remained stable in the silk nanofiber hydrogels. Doxorubicin (DOX) was effectively loaded on the vaterite spheres and silk nanofibers, forming synergistic silk-vaterite hydrogel delivery systems. The sustained delivery of DOX was tuned and controlled by vaterite stability and the DOX content loaded on the spheres and nanofibers. The cytotoxicity was regulated via the controlled delivery of DOX, suggesting the possibility of optimizing chemotherapeutic strategies. These silk-vaterite delivery hydrogels suggest a useful strategy for designing novel delivery systems for improved delivery and therapeutic benefits.