Reprogrammed Epigenetic Landscape-Prophesied Functions of Bioactive Polysaccharides in Alleviating Diseases: A Pilot Study of DNA Methylome Remodeling in Astragalus Polysaccharide (APS)-Improved Osteoporosis in a Rat Model.

DNA methylation is an epigenetic event that plays critical roles in the pathogenesis, progression, and treatment of human diseases. In this study, we investigated the epigenetic mechanisms for Astragalus polysaccharide (APS)-improved osteoporosis in a rat model. The results showed that APS significantly changed the DNA methylome in colonic epithelia with great efficiency. Gene set enrichment analysis (GSEA) based on differentially methylated sites (DMSs) revealed that APS caused promoter DNA methylation changes of genes associated with calcium homeostasis, osteoclast/osteoblast balance, Wnt signaling, and hormone-related processes. Further analysis showed high consistency of APS-induced gene methylomic changes in colonic epithelia and its effects on diabetes, virus infection, and wound healing, which had been reported already. Moreover, we suggested new functions and the involved mechanisms of APS in heart disease, neurological disorder, reproductive problem, and olfactory dysfunction. In this study, we offered epigenetic mechanisms for APS-improved osteoporosis. More importantly, we proposed and proved a reliable method to explore the beneficial effects of bioactive polysaccharides by studying DNA methylation changes at nonfocal sites. We firmly believed the promising prospects of this method for its great efficiency, rapidness, and economy in exploring possible beneficial or therapeutic effects of functional macromolecules with one single experiment.

Isolation and identification of iron-chelating peptides from casein hydrolysates.

Iron deficiency is a common nutritional disorder worldwide. Peptides derived from protein hydrolysates have recently attracted interest as novel iron chelators due to their superiority in terms of increasing solubility, bioavailability, absorption and stability. The aim of this study was to isolate and identify iron-chelating peptides from casein hydrolysates. Casein was hydrolyzed (trypsin, 3 h) and subsequently isolated using ultrafiltration and RP-HPLC. Four iron-chelating casein hydrolysate peptides, named CHP-1, CHP-2, CHP-3 and CHP-4, were identified by LC-MS/MS, and their amino acid sequences were Glu-Asp-Val-Pro-Ser-Glu-Arg (EDVPSER), His-Lys-Glu-Met-Pro-Phe-Pro-Lys (HKEMPFPK), Asn-Met-Ala-Ile-Asn-Pro-Ser-Lys (NMAINPSK) and Ala-Val-Pro-Tyr-Pro-Gln-Arg (AVPYPQR), with molecular weights of 830.6120 Da, 1012.5280 Da, 873.4440 Da and 829.4570 Da, respectively. The artificially synthesized peptides of CHP-1, CHP-2, CHP-3 and CHP-4 were verified, and their iron-chelating rates were 11.14%, 8.02%, 7.57% and 59.76%, respectively. These results suggested that the isolated iron-chelating peptides might serve as potential iron supplements and be used as food additives and functional foods.

Anti-fatigue and anti-oxidant activities of oyster (Ostrea rivularis) hydrolysate prepared by compound protease.

Oyster, which is rich in protein and widely used as a marine traditional Chinese medicine, was believed to have good curative effects in health care and on chronic diseases. This study was designed to evaluate the anti-fatigue and anti-oxidant effects of oyster hydrolysate. Oyster meat (OM) was hydrolyzed with a complex protease, and oyster hydrolysate (OH) was separated by a 6 kDa ultrafiltration membrane into two fractions, OH-I (<6 kDa) and OH-II (≥6 kDa). The anti-fatigue effects of OM, OH, OH-I and OH-II groups were first investigated, and then the antioxidant activities of OH-I and OH-II were further analyzed. Anti-fatigue experimental results showed that OH-I displayed the strongest activity among the four groups. Compared to the control group, OH-I significantly prolonged swimming time (67.79%), increased the content of muscle glycogen (45.65%) and liver glycogen (49.01%), and reduced the content of blood urea nitrogen (BUN) (18.44%) (P < 0.05). Meanwhile, OH-I showed excellent chemical and cellular antioxidant activities, especially when the concentration increased; its antioxidant activity was significantly better than that of OH-II (P < 0.05). Results of an amino acid analysis showed that OH-I was rich in branched-chain amino acids (10.84 g per 100 g), Glu (8.63 g per 100 g), Tau (1.68 g per 100 g), Asp (5.02 g per 100 g) and Arg (3.61 g per 100 g), which were expected to contribute to its antioxidant and anti-fatigue activities.

JAK2 inhibitor TG101348 overcomes erlotinib-resistance in non-small cell lung carcinoma cells with mutated EGF receptor.

Non-small cell lung cancer (NSCLC) patients with epidermal growth factor receptor (EGFR) mutations are responsive to EGFR-tyrosine kinase inhibitor (EGFR-TKI). However, NSCLC patients with secondary somatic EGFR mutations are resistant to EGFR-TKI treatment. In this study, we investigated the effect of TG101348 (a JAK2 inhibitor) on the tumor growth of erlotinib-resistant NSCLC cells. Cell proliferation, apoptosis, gene expression and tumor growth were evaluated by diphenyltetrazolium bromide (MTT) assay, flow cytometry, terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) staining, Western Blot and a xenograft mouse model, respectively. Results showed that erlotinib had a stronger impact on the induction of apoptosis in erlotinib-sensitive PC-9 cells but had a weaker effect on erlotinib-resistant H1975 and H1650 cells than TG101348. TG101348 significantly enhanced the cytotoxicity of erlotinib to erlotinib-resistant NSCLC cells, stimulated erlotinib-induced apoptosis and downregulated the expressions of EGFR, p-EGFR, p-STAT3, Bcl-xL and survivin in erlotinib-resistant NSCLC cells. Moreover, the combined treatment of TG101348 and erlotinib induced apoptosis, inhibited the activation of p-EGFR and p-STAT3, and inhibited tumor growth of erlotinib-resistant NSCLC cells in vivo. Our results indicate that TG101348 is a potential adjuvant for NSCLC patients during erlotinib treatment.

Enhancement of neutral endopeptidase activity in SK-N-SH cells by ginsenoside Rb1.

OBJECTIVES: To investigate the effect of ginsenosides Rb1 and Rg1 on Neprilysin (NEP) activity in SK-N-SH cells, and probe the underlying mechanism. METHODS: The effects of ginsenosides Rb1 and Rg1 on NEP activity were analyzed by NEP peptidase assay. Western blot was used to determine NEP gene expression at translational level, and RT-PCR was also performed to detect NEP gene expression at transcriptional level. RESULTS: NEP peptidase assay indicated that ginsenoside Rb1 can improve the activity of NEP, and RT-PCR and western blot results showed that the enhancement of NEP activity by ginsenoside Rb1 was due to enhancing NEP gene expression, while Rg1 did not have this effect. CONCLUSION: Our studies showed that ginsenoside Rb1 can enhance NEP activity by upregulating NEP gene expression. Our findings might offer a pharmacological explanation for the use of ginseng in traditional medicine.

Evaluation of transcutaneous electroacupoint stimulation with the train-of-four mode for preventing nausea and vomiting after laparoscopic cholecystectomy.

OBJECTIVE: To evaluate the efficacy of transcutaneous electroacupoint stimulation with a train-of-four (TOF) mode for the prevention of postoperative nausea and vomiting (PONV) in the patients undergoing laparoscopic cholecystectomy. METHODS: Ninety-six ASA Grade I - II patients scheduled for laparoscopic cholecystectomy were randomized into Neiguan (P6) electroacupoint stimulation group (treated group) and a placebo control group (placement of electrodes without electroacupoint stimulation). The anesthetic regimen was standardized by needling at Neiguan on the left side and connecting the TOF peripheral nerve stimulator. The incidence of nausea, vomiting, severity, antiemetic dosage and the degree of pain were assessed at 0, 60, 120 min, and 24 h after surgery. RESULTS: The incidence of nausea and vomiting, the dose of antiemetics and the occurrence of severe nausea were all significantly lower in the treated group compared with the control group and the score for pain was obviously reduced in patients of the treated group at 24 h post-operation (P<0.05 or P<0.01). CONCLUSION: Transcutaneous electroacupoint stimulation at P6 with the TOF mode could reduce the incidence and severity of nausea and vomiting with analgesic effects.