A biomimetic tri-phasic scaffold with spatiotemporal patterns of gastrodin to regulate hierarchical tissue-based vascular regeneration.

Clinical use of small-diameter vascular grafts remains a challenging issue in neovessel regeneration in view of thrombosis and intimal hyperplasia. Developing a vascular graft with structure and function similar to those of the native vessels necessitates a major direction of vascular tissue regeneration. Thus, this study sought to design and fabricate a range of tri-phasic scaffolds (0, 2, and 5 wt% gastrodin-polyurethane (PU)) with spatiotemporally defined structure and gastrodin-release for regulating the highly coordinated processes in growth of the intima and media. While the small pores of inner layer guided infiltration of human umbilical vein endothelial cells (HUVECs), the bigger pores of medial layer could offer smooth muscle cell (SMC)-friendly habitat, and external fibers conferred adequate mechanical properties. Correspondingly, spatial distribution and differential regulation of key proteins in HUVECs and SMCs were mediated by hierarchical release of gastrodin, of which rapid release in inner layer elicited enhanced HUVEC proliferation and migration against those of the SMC via activated endothelial nitric oxide synthase (eNOS) and heat shock protein 70 (HSP70) signal. Of note, superior anti-coagulation was reflected in 2 wt% gastrodin-PU ex vivo extracorporeal blood circulation experiment. After in vivo implantation for 12 weeks, there was no formation of obvious thrombosis and intimal hyperplasia in 2 wt% gastrodin-PU. The scaffold maintained high patency and improved vascular remodeling, including the formation of thin endothelialization in lumen and dense extracellular matrix deposition in medial layer. Taken together, the results demonstrate the positive function of hierarchical releasing system that responded to tri-phasic structure, which not only suppressed intimal thickening but also tightly controlled tissue regeneration.

Benefits of Huang Lian mediated by gut microbiota on HFD/STZ-induced type 2 diabetes mellitus in mice.

Background: Huang Lian (HL), one of the traditional Chinese medicines (TCMs) that contains multiple active components including berberine (BBR), has been used to treat symptoms associated with diabetes for thousands of years. Compared to the monomer of BBR, HL exerts a better glucose-lowering activity and plays different roles in regulating gut microbiota. However, it remains unclear what role the gut microbiota plays in the anti-diabetic activity of HL. Methods: In this study, a type 2 diabetes mellitus (T2DM) mouse model was induced with a six-week high-fat diet (HFD) and a one-time injection of streptozotocin (STZ, 75 mg/kg). One group of these mice was administrated HL (50 mg/kg) through oral gavage two weeks after HFD feeding commenced and continued for four weeks; the other mice were given distilled water as disease control. Comprehensive analyses of physiological indices related to glycolipid metabolism, gut microbiota, untargeted metabolome, and hepatic genes expression, function prediction by PICRUSt2 were performed to identify potential mechanism. Results: We found that HL, in addition to decreasing body fat accumulation, effectively improved insulin resistance by stimulating the hepatic insulin-mediated signaling pathway. In comparison with the control group, HL treatment constructed a distinct gut microbiota and bile acid (BA) profile. The HL-treated microbiota was dominated by bacteria belonging to Bacteroides and the Clostridium innocuum group, which were associated with BA metabolism. Based on the correlation analysis, the altered BAs were closely correlated with the improvement of T2DM-related markers. Conclusion: These results indicated that the anti-diabetic activity of HL was achieved, at least partly, by regulating the structure of the gut microbiota and the composition of BAs.

In vitro fermentation outcomes of arabinoxylan and galactoxyloglucan depend on fecal inoculum more than substrate chemistry.

Using in vitro fermentation conditions, this study investigated the fermentation characteristics of arabinoxylan (AX) and xyloglucan (XG) with a fecal inoculum that was collected either from humans consuming unrestricted diets or pigs fed a semi-defined diet with cellulose being the sole non-starch polysaccharide for 10 days prior to fecal collection. Metagenomic analysis revealed that microbial communities in the two types of inoculum were distinctively different, which led to distinct fermentation characteristics with the polysaccharides. The microbial communities fermented with the porcine fecal inoculum were clustered according to the fermentation time, while those fermented with the human fecal inoculum were differentiated by the substrates. Using the porcine fecal inoculum, irrespective of the substrates, Prevotella copri and the unclassified lineage rc4-4 were the dominant operational taxonomic units (OTUs) promoted during fermentation. Fermentation of wheat AX (WAX) and galacto-XG (GXG) with the human fecal inoculum, however, promoted different OTUs, except for a shared OTU belonging to Lachnospiraceae. Specifically, WAX promoted the growth of Bacteroides plebeius and a Blautia sp., while GXG promoted an unclassified Bacteroidales, Parabacteroides distasonis, Bacteroides uniformis and Bacteroides sp. 2. These changes in bacterial communities were in accordance with the short chain fatty acid (SCFA) production, where comparable SCFA profiles were obtained from the porcine fecal fermentation while different amounts and proportions of SCFA were acquired from fermentation of WAX and GXG with the human fecal inoculum. Altogether, this study indicated that the starting inoculum composition had a greater effect than polysaccharide chemistry in driving fermentation outcomes.

Extracellular depolymerisation triggers fermentation of tamarind xyloglucan and wheat arabinoxylan by a porcine faecal inoculum.

Arabinoxylan (AX) and xyloglucan (XG) are important components of primary cell walls of cereal grains and vegetables/fruits, respectively. Despite the established health benefits of these non-starch polysaccharides, the mechanisms of their utilisation by the gut microbiota are poorly understood. In this study, the mechanisms of solubilised wheat AX and tamarind XG degradation were investigated under in vitro fermentation conditions using a porcine faecal inoculum. Through structural analysis of the polymers, we demonstrate that depolymerisation by microbial surface accessible endo-degrading enzymes occurs prior to active fermentation of AX or XG. Breakdown products are released into the medium and potentially utilised cooperatively by other microbes. Acetate and propionate are the main fermentation products and are produced concurrently with polysaccharide depletion. Butyrate, however, is produced more slowly consistent with it being a secondary metabolite.

Mechanisms of utilisation of arabinoxylans by a porcine faecal inoculum: competition and co-operation.

Recent studies show that a single or small number of intestinal microbes can completely degrade complex carbohydrates. This suggests a drive towards competitive utilisation of dietary complex carbohydrates resulting in limited microbial diversity, at odds with the health benefits associated with a diverse microbiome. This study investigates the enzymatic metabolism of wheat and rye arabinoxylans (AX) using in vitro fermentation, with a porcine faecal inoculum. Through studying the activity of AX-degrading enzymes and the structural changes of residual AX during fermentation, we show that the AX-degrading enzymes are mainly cell-associated, which enables the microbes to utilise the AX competitively. However, potential for cross-feeding is also demonstrated to occur by two distinct mechanisms: (1) release of AX after partial degradation by cell-associated enzymes, and (2) release of enzymes during biomass turnover, indicative of co-operative AX degradation. This study provides a model for the combined competitive-co-operative utilisation of complex dietary carbohydrates by gut microorganisms.

[A primary study of immunotherapy with carcinoembryonic antigen peptide-pulsed, autologous human cultured dendritic cells in patients with advanced non-small cell lung cancer].

BACKGROUND: Dendritic cell (DC)-based immunotherapy is a new approach and effective for some malignant tumors. The aim of this study is to observe the efficacy and toxicity of immunotherapy with carcinoembryonic antigen (CEA) peptide-pulsed DCs in patients with refractory advanced lung cancer. METHODS: Lung cancer patients with high CEA expression were enrolled into this project. Autologous DCs were generated from patients' plastic-adherent peripheral blood mononuclear cells and loaded with CEA 5 days later. Cytokine-induced killer cells (CIK) were cultured from non-adherent peripheral blood mononuclear cells. DCs and CIK were transfused to patients. Responses and toxicities were observed. RESULTS: A total of 22 patients with lung cancer received DCs immunotherapy. DCs doses were 2.5×106-9.6×107 (5.03×106). CIK doses were 3.4×108-46×108. CD3, CD8, NK and IFN-γ levels obviously increased after treatment (P < 0.05). The 1-year survival rate was 68.2% (15/22). Main toxicities were fever and rash. CONCLUSIONS: DCs-based immunotherapy is feasible and safe to patients with lung cancer.