Characterization of exosomes derived from IPEC-J2 treated with probiotic Bacillus amyloliquefaciens SC06 and its regulation of macrophage functions.

Probiotics can maintain or improve health by modulating the response of immune cells in the gastrointestinal tract. However, the mechanisms by which probiotics promote macrophage (Mφ) activity are poorly understood. Here, we evaluated exosomes derived from intestinal epithelial cells treated with Bacillus amyloliquefaciens SC06 (Ba) and investigated the regulation of Mφ phagocytosis, apoptosis, and polarization. We isolated two exosomes from intestinal porcine epithelial cell lines (IPEC-J2) with or without Ba-treatment, named Ba-Exo and Exo, respectively. They had typical sizes and a cup-shaped morphology, and their surfaces presented typical exosomes-associated proteins, including CD63, ALIX, and TSG101. Ba-Exo and Exo could entrer Mφ (3D4/21 cells) effectively. Moreover, an in vitro phagocytosis assay demonstrated that Ba-Exo can promote phagocytosis of Mφ. Similar to Exo, Ba-Exo had no effect on Mφ apoptosis. Furthermore, Ba-Exo significantly increased inducible nitric oxide synthase (iNOS), declined the expression of arginase 1 (Arg1) in Mφ, and stimulated Mφ polarization to M1. To explore the differences in the regulation of Mφ polarization between Ba-Exo and Exo, we performed reverse transcription quantitative polymerase chain reaction analysis of the small RNAs and found that miR-222 increased in the Ba-Exo group compared to that in the Exo group. These results provide a new perspective on the relationship between probiotics and intestinal immunity.

Inhibition of glucuronomannan hexamer on the proliferation of lung cancer through binding with immunoglobulin G.

The anti-lung cancer activity of oligosaccharides derived from glucuronomannan was investigated. The inhibition of A549 cell proliferation by glucuronomannan (Gn) and its oligomers (dimer (G2), tetramer (G4) and hexamer (G6)) were concentration dependent. In vivo activities on the A549-derived tumor xenografts showed the tumor inhibition of G2, G4 and G6 were 17 %, 40 % and 46 %, respectively. Organ coefficients in nude mice showed an increase in the kidney with G4, the brain with G6, and the spleen with G6. An advanced tandem mass tag labeled proteomics approach was performed. A significant differential expression was found in 59 out of the 4371 proteins, which involved the immune system. Surface plasmon resonance (SPR) studies revealed G6 was strongly bound to immunoglobulin G. This suggests that glucuronomannan hexamer inhibits the proliferation of lung cancer through its binding to immunoglobulin.

Inhibitory effects of piceatannol on human cytomegalovirus (hCMV) in vitro.

Human cytomegalovirus (hCMV) is a ubiquitous herpesvirus, which results in the establishment of a latent infection that persists throughout the life of the host and can be reactivated when the immunity is low. Currently, there is no vaccine for hCMV infection, and the licensed antiviral drugs mainly target the viral enzymes and have obvious adverse reactions. Thus, it is important to search for compounds with anti-hCMV properties. The present study aimed to investigate the suppressive effects of piceatannol on hCMV Towne strain infection and the putative underlying mechanisms using human diploid fibroblast WI-38 cells. Piceatannol supplementation prevented the lytic changes induced by hCMV infection in WI-38 cells. Furthermore, piceatannol suppressed the expression of hCMV immediate-early (IE) and early (E) proteins as well as the replication of hCMV DNA in a dose-dependent manner. Moreover, hCMV-induced cellular senescence was suppressed by piceatannol, as shown by a decline in the senescence-associated ß-galactosidase (SA-ß-Gal) activity and decreased production of intracellular reactive oxygen species (ROS). p16INK4a, a major senescence-associated molecule, was dramatically elevated by current hCMV infection that was attenuated by pre-incubation with piceatannol in a dose-dependent manner. These results demonstrated that piceatannol suppressed the hCMV infection via inhibition of the activation of p16INK4a and cellular senescence induced by hCMV. Together, these findings indicate piceatannol as a novel and potent anti-hCMV agent with the potential to be developed as an effective treatment for chronic hCMV infection.

Sulfated galactofucan from Sargassum thunbergii induces senescence in human lung cancer A549 cells.

Isolated compounds from Sargassum thunbergii (S. thunbergii) have shown to exhibit diverse biological activities, including anti-cancer activity. In this study, we examined the effect of sulfated galactofucan (SWZ-4-H), which was successfully isolated from S. thunbergii, and its underlying mechanism on human lung cancer (LC) A549 cell growth in vitro and in vivo. In vitro experiment indicated that SWZ-4-H decreased cell growth and number in a dose-dependent manner (P < 0.05 vs. control). Besides, cells treated with SWZ-4-H had irregular morphology, including increased cell volumes, and large nuclei, which suggested senescence-like changes. Moreover, SWZ-4-H increased senescence-related ß-galactosidase (SA-ß-Gal) staining in a dose-dependent manner; however, while lower (1 mg mL-1) concentration induced mainly senescence without causing cell death, higher dosage (3 mg mL-1) induced both senescence and cell death. The effect of SWZ-4-H was further confirmed by analyzing the expression of p53, p21, p16, and Rb (p-RB); SWZ-4-H significantly increased the expression of p53, p21, and p16 and decreased phosphorylated Rb (p-RB) in a dose-dependent manner. Moreover, in vivo experiment showed that SWZ-4-H significantly reduced the tumor volume without affecting the body weight. To sum up, our data indicated that SWZ-4-H could induce lung cancer senescence by regulating p53, p21, p16, and p-Rb, thus providing a novel perspective on anti-cancer mechanisms of SWZ-4-H in human lung cancer A549 cells.

Protective effect of seleno-amino-oligosaccharide on oxidative damage of IPEC-1 cells by activating Keap1/Nrf2 signaling pathway.

The aim of the study was to investigate the antioxidant effect of seleno-amino-oligosaccharide (Se-AOS) on intestinal porcine epithelial cells (IPEC-1). MTT assay showed that Se-AOS had no effect on the viability of IPEC-1 cells up to a concentration of 9200 µg/L and Se-AOS significantly increased the viability of IPEC-1 cells compared to cells exposed to H2O2 alone. Se-AOS significantly increased the level of superoxide Dismutase (SOD) and decreased the levels of malonic dialdehyde (MDA) and lactate dehydrogenase (LDH) in IPEC-1 cells. The gene expression levels of different antioxidant enzymes dramatically increased by the pretreatment of Se-AOS compared to H2O2 treatment. In addition, the results indicated that Se-AOS up-regulated the intracellular Nrf2 and down-regulated the level of Keap1 by western blot. Taken together, these findings suggested that Se-AOS can protect IPEC-1 cells from oxidative damage through activating the Keap1/Nrf2 signaling pathway.

Aggressive behavior and brain neuronal activation in sexually naïve male Mongolian gerbils.

Aggressive behavior plays an important role in animal's survival and reproductive success. Although there has been growing interests in studying neural mechanisms underlying aggressive behavior using traditional laboratory animal models, little is known about mechanisms controlling naturally occurring aggression in sexually naïve animals. In the present study, we characterized aggressive behavior displayed by sexually naïve male Mongolian gerbils (Meriones unguiculatus) and examined the subsequent neuronal activation in the brain measured by Fos-immunoreactive (Fos-ir) staining. We found that resident males initiated attacks and showed intense levels of aggression (including chase, bite, offensive sideway, lunge and on-top) towards a conspecific male intruder. Furthermore, attacks from the resident males towards the intruder produced a nonrandom distribution of bites, with the most on the rump, flank, back and tail and few on the limbs, ventrum and head. In contrast, control males that were exposed to a woodblock (control for novelty) never attacked the woodblock and showed higher levels of object/environmental investigation. Male gerbils exposed to an intruder had significantly higher levels of Fos-ir density in the medial (MeA) and anterior cortical (ACo) subnuclei of the amygdala, principal nucleus (BSTpr) and interfascicular nucleus (BSTif) of the bed nucleus of the stria terminalis, ventrolateral subdivision of the ventromedial hypothalamus (VMHvl), and paraventricular nucleus of the hypothalamus (PVN), compared to control males. Together, our results indicate that sexually naïve, group housed male gerbils naturally display aggression towards conspecific strangers, and such aggressive behavior is associated with special patterns of neuronal activation in the brain.

Structure and Bioactivities of Porphyrans and Oligoporphyrans.

BACKGROUND: Pyropia (Porphyra), commonly known as nori or laver, is an important food source in many parts of the world. Edible dried Pyropia contains numerous nutrients and biofunctional components, including proteins, vitamins, eicosapentaenoic acid, minerals, carotenoids, mycosporine-like amino acids, and carbohydrate, and one of the compounds which we are interested in is porphyran, a sulfated polysaccharide comprising the hot-water-soluble portion of Pyropia cell walls. Researchers have performed a large number of in-depth studies on the biological activity and potential therapeutic applications of porphyrans and oligoporphyrans. METHODS: This mini review aims to provide comprehensive and update overview on the source, extraction, structure, biological activities and structure-activity relationships of porphyrans and oligoporphyrans based on the studies in the past 30 years which were included in Web of Science. RESULTS: The structure of porphyran has been basically determined given that its straight chain is relatively simple, and the skeleton structure has been described. The extraction methods were simplified continuously, but different extraction methods and post- processing methods still had great influence on the structure and composition of porphyran, so there was no standardized extraction process which can achieve quality control until now. In order to obtain oligoporphyrans, there are a variety of degradation methods, including chemical method, physical method and enzymatic method, but it is worth mentioning that specific degradation enzyme is still unavailable. Studies on the biological and pharmacology properties include antioxidant, anti-tumor, anti-inflammatory, immunomodulation, anti-cardiovascular and cerebrovascular diseases and drug delivery. CONCLUSION: Owing to the therapeutic potential and drug delivery applications, porphyran and oligoporphyrans are expected to be further developed as a medicine against human diseases, as well as a supplement in cosmetics and health products.

Synthesized sulfated and acetylated derivatives of polysaccharide extracted from Gracilariopsis lemaneiformis and their potential antioxidant and immunological activity.

The sulfated and acetylated derivatives of the sulfated polysaccharide extracted from red algae Gracilariopsis lemaneiformis were synthesized, and their antioxidant and immunological activities were investigated. And then the relationship between the activity and chemical characteristics was characterized. The results of chemical analysis and FT-IR showed the modification of polysaccharide was successful. In addition, it was found that a certain derivative exhibited stronger antioxidant activity than the one unmodified in the same experiment. The sulfated and acetylated derivatives showed different activities on the T and B lymphocytes proliferation. They may be of potential value as a possible supplement in food and pharmaceutical industries.

Elevated pretreatment platelet distribution width and platelet count predict poor prognosis in nasopharyngeal carcinoma.

BACKGROUND: Previous studies have demonstrated that platelets play a multifaceted role in cancer progression and metastasis. However, the value of platelet indices for predicting survival in nasopharyngeal carcinoma (NPC) patients remains unknown. The aim of this study was to evaluate the predictive significance of platelet indices in NPC cases. MATERIALS AND METHODS: A total of 168 patients who were diagnosed with NPC between January 2011 and June 2012 were recruited. The optimal cut-off values for the platelet indices were determined using a receiver operating characteristic curve. The Kaplan-Meier method and Cox regression were used to evaluate the prognostic impact of the potential predictors. RESULTS: Of the 168 patients, high platelet distribution width (PDW) and platelet count (PLT) levels were observed in 81 (48.21%) and 68 (40.48%) of the patients, respectively. An increased PDW was associated with the depth of invasion (T stage, P = 0.019), lymph node metastasis (N stage, P = 0.026), and clinical stage (P < 0.001). Moreover, the survival analysis showed that the overall survival of the patients with a PDW > 16.3 fL or platelet count > 266 × 109/L was associated with a poorer prognosis (both P < 0.001). In the multivariate Cox regression model, the PDW (P < 0.001), PLT (P = 0.001), T stage (P < 0.001), N stage (P = 0.006), clinical stage (P = 0.005), and Epstein-Barr virus DNA (P = 0.039) were independent prognostic factors for the overall survival. CONCLUSIONS: The PDW and PLT are easily available via a routine blood test, and our study showed that the PDW and PLT could be prognostic predictors in NPC patients. However, further studies are required to confirm this conclusion.

The antitumor activity of a red alga polysaccharide complexes carrying 5-fluorouracil.

Porphyran is a sulfated galactan isolated from red algae Porphyra haitanensis, and have been reported to have many kinds of biological activities such as antitumor activity. In order to provide a water-soluble macromolecule prodrug of 5-Fu showing slow release of 5-Fu and reducing side-effect, we carried out fixation of 5-Fu to porphyran at 6-position. In this study, the antitumor and immunomodulation activities of low MW porphyran carrying 5-Fu on transplanted S180 tumor mice were studied. Weight of immune organ, proliferation ratio of lymphocyte concentration of TNF-α and NO from the transplanted S180 tumor mice were also determined. Results indicated that the conjugate could enhance antitumor activity of 5-Fu and improve immunocompetence damaged by 5-Fu.

The immunological and antioxidant activities of polysaccharides extracted from Enteromorpha linza.

Polysaccharides extracted with water and alkali from Enteromorpha linza were a group of hetero polysaccharides, and their immunological and antioxidant activities were investigated employing various established in vitro systems. The structural analysis showed that they were composed of rhamnose, mannose, xylose, glucose, and galactose with different molar ratio. Available data obtained with in vitro models suggested that the two kinds of polysaccharide showed significant inhibitory effects on radical and could increase the capability of T and B lymphocytes proliferation. These results clearly establish the possibility that these polysaccharides extracted from E. linza could be effectively employed as ingredient in health or functional food, to all eviate oxidative stress. However, comprehensive studies need to be conducted in experimental animal models.

Physicochemical properties and inhibition effect on iron deficiency anemia of a novel polysaccharide-iron complex (LPPC).

Porphyran (P) was extracted from red algae Porphyra by boiling water. A novel polysaccharide-iron complex (LPPC) was prepared under the alkaline condition by adding a ferric chloride solution to the low molecular weight porphyran (LP) solution. Physicochemical properties and inhibition effect on iron deficiency anemia of this complex were studied. The content of iron(III) in the complex is 21.57% determined with iodometry. The results indicate that LPPC was product required. The complex can increase red blood cell count (RBC), hemoglobin (Hb), Serum iron (SI), spleen index, spleen mass and mass of mice with iron deficiency anemia (IDA). Although the structure and deeper mechanisms on hemolytic anemia of LPPC should be further studied, LPPC is hoped to be developed as a late-model iron supplement which has a synergism on anemia.

Isolation and antioxidant activities of polysaccharides extracted from the shoots of Phyllostachys edulis (Carr.).

Polysaccharides extracted from Phyllostachys edulis (Carr.) are a group of hetero polysaccharides, and their antioxidant activities were investigated employing various established in vitro systems. Available data obtained with in vitro models suggested that among the three samples, B1 (extraction with water) showed significant inhibitory effects on superoxide radical and hydroxyl radical; its reducing power was also the strongest among the three samples. These results clearly establish the possibility that polysaccharides extracted from P. edulis could be effectively employed as ingredient in health or functional food, to alleviate oxidative stress. However, comprehensive studies need to be conducted in experimental animal models.

Synthesized phosphorylated and aminated derivatives of fucoidan and their potential antioxidant activity in vitro.

Three sulfated polysaccharide derivatives (phosphorylated and aminated fucoidan) were synthesized, and their potential antioxidant activities were investigated employing various established in vitro systems. Two methods were used in phosphorylation fucoidan: polyphosphoric acid and POCl(3) method. Aminated fucoidan was prepared using the epichlorohydrin and ammonia water. All fucoidan derivatives possessed considerable antioxidant activity, and exhibited stronger antioxidant ability than fucoidan in certain tests. The phosphorylated fucoidan showed stronger hydroxyl radical and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and reducing power. The mechanism on influence the antioxidant activity of samples of phosphate and amino group was indicated.