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Background Pemphigus is an organ-specific autoimmune bullous disease. Objectives To determine the role of regulatory B cells (Bregs) in patients with pemphigus. Methods The frequency of the occurrence of CD19+ CD24hi CD38hi Bregs was detected from 34 patients with pemphigus and 20 healthy controls. Interleukin ( IL)-10 secretion was processed after stimulating B cells. Specific antidesmoglein antibody (Ab) titres and their subclasses were also measured. Ab response and cytokine production from peripheral blood mononuclear cells ( PBMCs) with or without Bregs were analysed. Results The number of Bregs was significantly increased in patients with pemphigus compared with healthy controls (15 ± 7% vs. 9 ± 3%; P < 0·01) and the proportion of Bregs in the active groups (newly diagnosed and chronic active patients) was significantly higher than in remittent individuals (16 ± 7% vs. 13 ± 8%; P = 0·04). The IL-10-producing B cells were significantly increased upon stimulation both in patients and in healthy controls. However, the increase ratio of IL-10-producing B cells between short- and long-term stimulation was significantly lower in patients with pemphigus (1·0-fold vs. 2·6-fold increase in control group; P < 0·01). Strikingly, Bregs from the controls were able to suppress interferon (IFN)-γ expression and T helper cell 1 (Th1) immune response (26% inhibition rate), while the suppressive function of Bregs from patients with pemphigus was significantly decreased (9% inhibition rate). There was no difference in Ab levels from PBMCs with or without Bregs after stimulation. Conclusions Bregs in patients with pemphigus are elevated but with defective regulatory function on Th1 cells. [ABSTRACT FROM AUTHOR]

Dentin bonding is a dynamic process that involves the penetration of adhesive resin monomers into the extrafibrillar and intrafibrillar demineralized collagen matrix using a wet-bonding technique. However, adhesive resin monomers lack the capacity to infiltrate the intrafibrillar space, and the excess water that is introduced by the wet-bonding technique remains at the bonding interface. This imperfectly bonded interface is inclined to hydrolytic degradation, severely jeopardizing the longevity of bonded clinical restorations. The present study introduces a dentin bonding scheme based on a dry-bonding technique, combined with the use of extrafibrillar demineralization and a collagen-reactive monomer (CRM)-based adhesive (CBA). Selective extrafibrillar demineralization was achieved using 1-wt% high-molecular weight (MW) carboxymethyl chitosan (CMCS) within a clinically acceptable timeframe to create a less aggressive bonding substance for dentin bonding due to its selectively extrafibrillar demineralization capacity. CMCS demineralization decreased the activation of in situ collagenase, improved the shrinking resistance of demineralized collagen, and thus provided stronger and more durable bonding than traditional phosphoric acid etching. The new dentin bonding scheme that contained CMCS and CBA and used a dry-bonding technique achieved an encouraging dentin bonding strength and durability with low technical sensitivity. This bonding scheme can be used to improve the stability of the resin-dentin interface and foster the longevity of bonded clinical restorations., Competing Interests: The authors declare no competing interest., (© 2021 The Authors.)

Objective: To investigate the effect of Na(+)/K(+)-ATPase inhibitor ouabain on the proliferation and division of liver cancer HepG2 cells, and to explore the anticancer mechanism. Methods: HepG2 cells were exposed with different concentrations of ouabain (0.1, 1, 10 μmol/L) for 24 h, the proliferation ability was appraised using CCK-8, and the HepG2 cells was as a control group. The status of chromosome separation was detected with cell immunofluorescence (ICC) coupled to confocal microscope. The expression levels of AURKA, mTOR, p-mTOR, ERK and p-ERK protein were analyzed using western blot. Results: After treating with 0.1, 1 and 10 μmol/L of ouabain for 24 h, the inhibitory rate of cells were (23.5±4.57)%, (49.80±5.32)%, and (72.10±5.62)%, respectively. Ouabain could significantly inhibit the proliferation of HepG2, and presented in a dose-dependent manner( F= 32.8, P< 0.05). The ICC results showed that the chromosome separation disorders occurred in HepG2 cells treated with 1 μmol/L for 24 h, and the spindle diameter of HepG2 cells with ouabain treatment was decreased significantly compared with the control group( t= 9.58, P< 0.05). The results of western blot showed that the expression levels of AURKA, p-mTOR and p-ERK expressions in HepG2 cells treated with 1 μmol/L of ouabain were significantly decreased compared with the control group( F= 16.26, 8.32, 33.59, P< 0.05). Ouabain inhibited the growth of hepatocellular carcinoma cells in nude mice( F= 370.20, P< 0.05). Conclusion: Ouabain can induce chromosome division disorder and inhibit the proliferation in liver cancer HepG2 cells by inhibiting AURKA signaling pathway.

This study evaluated the use of a new collagen-reactive monomer (CRM), isocyanate-terminated urethane methacrylate precursor, which has covalent affinity to dental collagen, in the formation of dentin-resin bonds and compared it with 2 other dental adhesives. Dentin specimens were bonded with either the CRM-based adhesive (CBA), One-Step (OS; Bisco, Inc.), or a negative adhesive (NA) control and subjected to 24-h storage in water, thermocycling to simulate 1-y clinical function, or a matrix metalloproteinase-mediated aging process. We tested the microtensile bond strength (µTBS), characterized the bonding interface with an atomic force microscope, conducted micro-Raman analysis, and performed leakage tests and in situ zymography. CBA and OS exhibited comparable bonding strength after 24 h ( P > 0.05); however, there was a sharp decrease in µTBS after aging for all except CBA ( P < 0.001). Raman spectra results indicated increased collagen crosslinking and chemical reaction between the adhesive and collagen in the CBA group. CBA achieved high-quality hybridization with collagen, improving mechanical properties and integrity, and decreased the enzyme-mediated degradation of the bonding interface by inhibiting collagenolytic activity. With the promising bonding durability of coapplied CBA, CRM may be the first dental adhesive to provide strong and long-lasting resin-dental collagen bonding without the additional conditioning step. The use of CBA results in high-quality hybrid layers that protect the resin-dentin interface from harmful biological and chemical activities commonly occurring in the oral environment.

When damaged or fractured collagen-rich hard tissues are repaired by resin material, the collagen matrix may be used as a scaffold, after removal of the natural minerals, for resin monomers to penetrate and polymerize in-situ. Formation of a collagen-polymer hybrid biocomposite via mechanical hybridization provides a stable and strong link between endogenous tissue and the prosthesis for successful clinical integration. However, the heterogeneity between hydrophobic resin polymers and hydrophilic collagen presents a challenge to the quality of hybrid biocomposite. The objective of the present study was to evaluate the potential benefits of a collagen-reactive monomer (CRM, an isocyanate-terminated urethane-based methacrylate) with covalent affinity to collagen as "chemical link" to enhance in-situ resin hybridization within a collagen scaffold. Here, the CRM ligand with active isocyanate group may be chemically grafted onto the collagen receptor via covalent and hydrogen bonds. Dentin-derived collagen chemical modified by CRM shows improved mechanical property, thermostability and enzymatic stability. Moreover, CRM inhibited both exogenous and endogenous collagenase activities. The modification of collagen by chemical grafting of resin monomers improved its mechanical and physicochemical properties and demonstrated the potential of CRM for use in promoting chemical adhesion and creating a much stronger and durable bonding interface. Formation of a chemical bond between polymer and collagen scaffold in-situ improves the mechanical performance of collagen and may create a much stronger and durable collagen-polymer hybrid material. Addition of CRM into adhesives might effectively prolong the longevity of clinical resin-bonded restorations., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Pemphigus is an organ-specific autoimmune bullous disease., Objectives: To determine the role of regulatory B cells (Bregs) in patients with pemphigus., Methods: The frequency of the occurrence of CD19(+) CD24(hi) CD38(hi) Bregs was detected from 34 patients with pemphigus and 20 healthy controls. Interleukin (IL)-10 secretion was processed after stimulating B cells. Specific antidesmoglein antibody (Ab) titres and their subclasses were also measured. Ab response and cytokine production from peripheral blood mononuclear cells (PBMCs) with or without Bregs were analysed., Results: The number of Bregs was significantly increased in patients with pemphigus compared with healthy controls (15 ± 7% vs. 9 ± 3%; P < 0·01) and the proportion of Bregs in the active groups (newly diagnosed and chronic active patients) was significantly higher than in remittent individuals (16 ± 7% vs. 13 ± 8%; P = 0·04). The IL-10-producing B cells were significantly increased upon stimulation both in patients and in healthy controls. However, the increase ratio of IL-10-producing B cells between short- and long-term stimulation was significantly lower in patients with pemphigus (1·0-fold vs. 2·6-fold increase in control group; P < 0·01). Strikingly, Bregs from the controls were able to suppress interferon (IFN)-γ expression and T helper cell 1 (Th1) immune response (26% inhibition rate), while the suppressive function of Bregs from patients with pemphigus was significantly decreased (9% inhibition rate). There was no difference in Ab levels from PBMCs with or without Bregs after stimulation., Conclusions: Bregs in patients with pemphigus are elevated but with defective regulatory function on Th1 cells., (© 2014 British Association of Dermatologists.)

Objective: To study the mechanism of inhibiting actions of bufalin on human leukemic cells (HL60)., Methods: HL60 cells were incubated with bufalin at different concentrations and growth inhibition was analyzed by trypan-blue staining. Cell apoptosis was evaluated by phase contrast microscopy, transmission eletron microscopy and agarose gel electrophoresis., Results: Bufalin remarkably inhibited the growth of HL60 cells. Apoptosis of HL60 cells could be effectively induced by bufalin at concentration of 0.01 mumol.L-1 or higher, showing the apoptotic changes in morphology, including cell shrinkage, chromatin agglutination and formation of apoptotic corpuscula; DNA cleavage was observed with agarose gel electrophoresis., Conclusion: Bufalin can effectively induce apoptosis of HL60 cells, which is possibly one of the mechanisms for anti-cancer of bufalin.