ABSTRACT
@#Introduction: Dental caries in children is a major problem of mouth disease throughout the world, so too is there currently an increase in health problems in children due to obesity. Human Beta defensing(HBD) has been found in saliva and from several studies stated that HBD aside from being a broad-spectrum antimicrobial can act as an immunomodulator. The purpose of this study is to reveal whether there is a relationship between obesity and HBD-3 salivary concentration in caries patients and caries-free patients. Methods: This cross-sectional observational study was involved 62 children with caries and caries-free, aged 9-11 years, students at Qommarudin Islamic Boarding School, Gresik, East Java Indonesia. dental caries examination, carried out in accordance with World Health Organization (WHO) diagnostic criteria. Body mass index (BMI) was measured from the height and weight of individuals, HBD3 concentrations were tested with an ELISA kit from Bioassay Technology Laboratory (China) from saliva samples. Evaluate the results with the Kruskal Wallis test, followed by the Mann-Whitney test. The level of significance used in this statistical test was 0.05. Results: there was a relationship between BMI level and HBD-3 concentration in the caries group (p <0.05, p = 0.009) with a moderate level of association. but there was no significant relationship in the caries-free group (p> 0.05, p = 0.189). Conclusion: There was an association between BMI and HBD-3 salivary concentration in caries patients but there was no relationship in the caries-free group.
ABSTRACT
PURPOSE: To investigate whether secretion of human beta-defensin 3 (HBD-3) is induced by bacillus Calmette-Guerin (BCG) and to determine whether HBD-3 affects BCG internalization in bladder cancer cells. MATERIALS AND METHODS: RTPCR analysis was used to determine whether HBD-3 mRNA increases after incubation with BCG. HBD-3 proteins in 5637 and T24 human bladder cancer cell lines were assayed by ELISA. The internalization rate was evaluated by double immunofluorescence assay and confocal microscopy to test the optimal dose of HBD-3 for BCG internalization. We also investigated the difference in internalization rates and cell viability between recombinant HBD-3 protein, anti-HBD-3 antibody, and HBD-3 plus anti-HBD-3 antibody pretreatments. RESULTS: BCG induced HBD-3 mRNA expression and HBD-3 production dose and time-dependently in bladder cancer cells and affected BCG internalization. Pretreatment with recombinant HBD-3 protein lowered internalization of BCG dose-dependently. Moreover, anti-HBD-3 antibody prevented the effect of HBD-3 on BCG internalization in bladder cancer cells. The internalization rate of BCG pretreated with anti-HBD-3 antibody was higher than that in the control. The BCG internalization rate in cells pretreated with anti-HBD-3 antibody plus recombinant HBD-3 protein was higher than that in the control. BCG decreased bladder cancer cell viability, and anti-HBD-3 antibody prevented the inhibitory role of HBD-3 on the anti-proliferative effects of M. bovis BCG in bladder cancer cells. CONCLUSIONS: Bladder cancer cells produce HBD-3 when they are infected by BCG to defend themselves against BCG internalization, which plays an important role during the initiation and propagation of the immunotherapeutic response in bladder cancer cells.
Subject(s)
Humans , Bacillus , Cell Line , Cell Survival , Enzyme-Linked Immunosorbent Assay , Fluorescent Antibody Technique , Microscopy, Confocal , Mycobacterium bovis , RNA, Messenger , Urinary Bladder Neoplasms , Urinary BladderABSTRACT
Objective To achieve the fusion expression of the entire human beta-defensin-3(hBD-3) gene. Method We synthesized two oligonucleotide primers accor ding to the codon preference of Escherichia coli. The gene was cloned into p GEX -4T-2 to establish the pGEX-4T-2-hBD-3 as the fusion expression vector by PCR. Transformed into E.coli strain DH5α, the express vector was induced an d ex pressed by IPTG. The fusion protein GST-hBD-3 was obtained by repeated cycles of freezing and thawing, cut by thrombin to attain the recombinant hBD-3 protei n. Result The result of the antibacterial peptide agarose diffu sion assay shows the antibacterial activity of the rhBD-3 against the S.aureu s exists, and it reaches 0.843U. Conclusion The fusion expr ession of the hBD-3 gene is successful.