Separation of bufadienolides from Helleborus thibetanus Franch. by a combination approach involving macroporous resin column chromatography and gradient countercurrent chromatography.

In this study, a combination approach involving macroporous resin (MR) column chromatography and gradient countercurrent chromatography (CCC) was employed to enrich and purify bufadienolides from the roots and rhizomes of Helleborus thibetanus Franch. Initially, a D101 MR-packed column chromatography was utilized for fractionation and enrichment of the bufadienolides, which were effectively eluted from the column using a 60% ethanol solution. CCC was subsequently introduced to separate the enriched product using the ethyl acetate/n-butanol/water (EBuWat, 4:1:5, v/v) and EBuWat (5:0:5, v/v) solvent systems in a gradient elution mode. As results, five bufadienolides, including 6.1 mg of hellebrigenin-3-O-ß-D-glucoside (1), 2.2 mg of tigencaoside A (2), 8.3 mg of deglucohellebrin (3), 3.5 mg of 14 ß-hydroxy-3ß-[ß-D-glucopyranosyl-(1→6)-(ß-D-glucopyranosyl)oxy]-5α-bufa-20,22-dienolide (4), and 3.0 mg of 14ß-hydroxy-3ß-[(ß-D-glucopyranosyl)oxy]-5α-bufa-20,22-dienolide (5), were effectively separated from 300 mg of the enriched product. The respective high-performance liquid chromatography purities were as follows: 95.2%, 75.8%, 85.7%, 82.3%, and 92.8%. This study provides valuable insights for the efficient enrichment and separation of bufadienolides from Helleborus thibetanus Franch.

ε­Polylysine-stabilized agarose/polydopamine hydrogel dressings with robust photothermal property for wound healing.

Polydopamine (PDA) is emerging as an attractive photothermal agent due to its good photothermal performance and excellent biocompatibility. However, without chemical modification, PDA is normally unstable and usually leached out from the constructed biomaterials, realistically limiting its application space. Here, we constructed a new hydrogel dressing with robust and stable photothermal performance by introduction of ε-Polylysine (ε-PL) into agarose/PDA matrix to efficiently lock PDA. By optimizing PDA/ε-PL rational dose in agarose network structure, a hybrid agarose/PDA/ε-PL hydrogel (ADPH) with stable photothermal functionality and desirable physicochemical properties could be achieved. ADPH possessed satisfactory microbicidal efficacy in vivo, which enabled the bacteria-infected skin wound to be cured quickly by successful suppressing inflammation, accelerating collagen deposition and promoting angiogenesis in a bacterial-infected wound model. Collectively, this study illustrates a simple, convenient but powerful strategy to design functionally stable ADPH dressing for treating dermal wounds, which could open vistas in clinical wound management.

[Temporal expression of PD-1 and PD-L1 during the development of experimental periodontitis in rats and its implications].

PURPOSE: To study the temporal expression of programmed death receptor 1 (PD-1) and its ligand (PD-L1) in the development of periodontitis in rats. METHODS: SD rats were randomly divided into control group and model group, and 4 subgroups were divided in each model group according to the time of measurement: group A (1 week), group B (2 weeks), group C (3 weeks) and group D (4 weeks). There were 8 rats in each subgroup. Maxillary periodontitis models were made by using "thread ligation + vaccination LPS-PG" in rats. Periodontal tissue specimens were examined and bone resorption areas were determined in each group. Tumor necrosis factor alpha (TNF-α), interleukin-1 (IL-1ß), interleukin 6 (IL-6) and transforming growth factor beta (TGF-ß) mRNA in periodontal tissue in each group were determined by RT-PCR method. PD-1 and PD-L1 protein expression in periodontal tissues in each group were determined by Western blotting. The data were analyzed by SPSS 22.0 software package. RESULTS: During modeling period, amelocemental junction-alveolar crest(ACJ-AC) distance and bone resorption area of the first molar in model group gradually increased (P<0.05), which were significantly different from the control group at the corresponding time point (P<0.01). During modeling period, TNF-α, IL-1ß and IL-6 mRNA level in periodontal tissues in the model group was continuously increased(P<0.05), and TGF-ß mRNA was continuously decreased(P<0.05), which was significantly different from the control group at the corresponding time point(P<0.01). During disease progress, PD-1 and PD-L1 protein level in periodontal tissues in each model group was continuously increased(P<0.05), which was significantly different from the control group at the corresponding time point (P<0.01); and PD-1 and PD-L1 protein levels in periodontal tissues in each model group was positively correlated to TNF-α and IL-6 mRNA levels(P<0.01). CONCLUSIONS: PD-1, as an immunosuppressive molecule and its receptor PD-L1, can promote the progression of periodontal inflammation, and its effect may be achieved by regulating the expression of TNF-α and IL-6.Regulating the expression of PD-1 and PD-L1 may be a new target for the treatment of periodontitis.

Defect of cell wall construction may shield oral bacteria's survival in bloodstream and cause infective endocarditis.

Infective endocarditis (IE) is a rare but life-threatening infection. Bacteremia with organisms known to cause IE occurs commonly in association with invasive dental origin. Despite daily oral activities as well as professional dental treatments inducing bacteremia and the dental bacteremia as a risk factor of IE, the details of dental bacteria in the pathogenesis of IE are far from elucidation to date. How do a few microorganisms survive host defenses or escape from antibiotic attacking to seed target organs and cause distant infections? Why are Gram-positive bacteria more frequently detected than Gram-negative bacteria in IE? Cell wall-deficient bacteria (CWDB) were traditionally defined as bacteria with altered morphology and consistent with damaged or absent cell wall structures identified by EM. A number of case reports and laboratory studies suggest that CWDB may be found in the peripheral blood of patients with IE, and may also be demonstrated in vegetations on the valves of patients with IE. CWDB, in vitro, are resistant to antibiotics that act on cell wall biosynthesis. Recent studies indicate that the Streptococcus mutans (S. mutans) strains, the major cariogenic bacterium, isolated from the infected valve were deficient in some wall-associated proteins which are main cariogenic virulence of S. mutans, and the deficient stains exhibited less susceptible to antibiotics that act on cell wall biosynthesis. Further, the cloned deficient mutans were less susceptible to phagocytosis by human polymorphonuclear leukocytes but to possess higher platelet aggregation properties than their parent strains. As outlined above, we hypothesize that defect of cell wall construction may shield oral bacteria's survival in bloodstream and cause IE.