The level of antimicrobial peptides in gingival crevicular fluid and its correlation with periodontal clinical indexes in elderly patients with type 2 diabetic periodontitis / 口腔疾病防治

Objective@#To investigate the changes and significance of human beta-defensin-2 (HBD-2) and LL-37 in the gingival crevicular fluid of patients with periodontitis and type 2 diabetes mellitus (T2DM).@*Methods@#This study was conducted among 45- to 85-year-old patients in the Department of Stomatology and Internal Medicine of Shenzhen Center for Chronic Disease Control, including a healthy control group of 22 people, a systemically healthy control group of 19 people with periodontitis, a T2DM periodontal health group of 15 people, and a T2DM group of 21 people with periodontitis. The Florida periodontal probe was used for periodontal examination, and the clinical indexes, including probing depth (PD), clinical attachment level (CAL) and probing on bleeding (BOP), were recorded. The concentrations of HBD-2 and Ll-37 in gingival crevicular fluid were determined by ELISA. The differences in HBD-2, LL-37 and periodontal clinical indexes between the groups were compared, and correlation analysis was conducted.@*Results@#The PD values in T2DM with the periodontitis group were higher than those of the systemically healthy controls with periodontitis group (P < 0.05); the levels of HBD-2 and LL-37 in gingival crevicular fluid in systemically healthy controls with periodontitis group were significantly higher than those in the healthy control group (P < 0.05), the level of HBD-2 in gingival crevicular fluid in systemically healthy controls with periodontitis group was significantly higher than that in T2DM with periodontitis group (P < 0.05); and the antimicrobial peptides HBD-2 and LL-37 in gingival crevicular fluid were significantly positively correlated with the PD and CAL in systemically healthy controls with periodontitis group (P < 0.05), and there was no significant correlation between the antimicrobial peptides HBD-2, LL-37 in gingival crevicular fluid and PD, CAL in T2DM with periodontitis group (P > 0.05).@*Conclusion@#The levels of antimicrobial peptides HBD-2 and LL-37 in gingival crevicular fluid of middle-aged and elderly patients with T2DM periodontitis were lower, and there was no significant correlation with PD and CAL in periodontal clinical indicators.

TLR4/NF-κB signal pathway mediate HBD-2 in patients with preterm premature rupture of membrane associated with histologic chorioamnionitis / 中国综合临床

Objective To observe the effect of human β-defensins-2(HBD-2) for chorioamnionitis(HCA) pregnant women before term premature rupture of fetal membrane(PROM) process,and explore toll-like receptor 4 / nuclear factor-κ B (TLR4 / NF-κB) predominate role in the process of signal transduction pathway in the mechanism.Methods Fifty five women with PROM were enrolled in the study.According to the Results of pathological diagnosis of membranes,pregnant women with PROM divided into histological chorioamnionitis,HCA and non-HCA.The same sample without PROM pregnancies matching the same gestational ages were recruited as control group.We examined the messenger RNA(mRNA) of TLR4,NF-κB p65 and HBD-2 in placenta and fetal membrane real-time reverse transcription polymerase chain reactions by dsDNA-binding dyes of SYBR Green.Results (1)In the placenta,the level of TLR-4(17.15±4.52),NF-κB p65(47.11±14.23),HBD-2mRNA(27.35±2.67) in PROM group were significantly higher than the level of TLR-4(7.21±3.25),NF-κB p65(30.51±13.05),HBD-2mRNA(13.55±0.8) in control group(t=-1.966,-1.474,-1.754,P0.05).Conclusion Linear positive correlation of TLR4,NF-κB and HBD2 indicated that TLR4/NF-κB/HBD2 signal transduction pathway may be involved in the development of preterm premature rupture of membrane associated with histologic chorioamnionitis.

Construction and identification of recombinant lentivirus carrying human beta defensins-3 and connective tissue growth factor gene / 军事医学

Objective To construct a recombinant lentivirus containing human beta defensins -3 ( hBD3 ) , connective tissue growth factor gene (CTGF) and enhanced green fluorescent protein (EGFP), and to detect its translation in rabbit bone marrow mesenchymal stem cells (BMSC).Methods The lentivirus containing hBD3, CTGF and EGFP genes was constructed in vitro.The titer of lentivirus was tested with end-paint dilution assay .Rabbit BMSCs were transfected with recombinant virus.The best value of multiplicity of infection (MOI) was tested.The expression condition, transfection efficacy and genetic stability of the target genes were evaluated by using fluorescence microscopy and flow cytometry . Western blotting was used to detect the expression of the target protein .Results Recombinant lentivirus vectors: Lenti-CTGF-hBD3-EGFP, Lenti-hBD3-EGFP, and Lenti-EGFP, were successfully obtained . The titer of the recombinant lentiviruses was 3.21 ×108, 5.80 ×108, and 1.16 ×109, respectively.The best MOI value to transfect BMSCs was 150. The transfection efficacy of these lentivirus vectors was high , reaching 79.72%as assessed by flow cytometry , and it could be stably inherited .Western blotting displayed that target protein expression was successful .Conclusion The construction of recombinant lentiviruses carrying hBD3 and CTGF genes is successful and can be effectively transfected into BMSCs .

Expression of TIM-3, Human beta-defensin-2, and FOXP3 and Correlation with Disease Activity in Pediatric Crohn's Disease with Infliximab Therapy

BACKGROUND/AIMS: This study investigated the expression of T cell immunoglobulin- and mucin-domain-containing molecule 3 (TIM-3), human beta-defensin (HBD)-2, forkhead box protein 3 (FOXP3), and the frequency of CD4+ CD25+ FOXP3+ regulatory T cells (Tregs) in children with Crohn's disease (CD) during infliximab therapy. METHODS: We enrolled 20 CD patients who received infliximab treatment for 1 year. Peripheral blood and colonic mucosal specimens were collected from all CD patients and from healthy control individuals. RESULTS: A significant difference in TIM-3 mRNA expression was evident in peripheral blood mononuclear cells and colonic mucosa between CD patients before infliximab therapy and the healthy controls (p<0.001 and p=0.005, respectively). A significant difference in HBD-2 mRNA expression was found in colonic mucosa between CD patients before infliximab therapy and the healthy controls (p=0.013). In the active phase of CD, at baseline, the median percentage of T cells that were CD25+ FOXP3+ was 1.5% (range, 0.32% to 3.49%), which increased after inflixmab treatment for 1 year to 2.2% (range, 0.54% to 5.02%) (p=0.008). CONCLUSIONS: Our study suggests that both the adaptive and innate immune systems are closely linked to each other in CD pathogenesis. And the results of our study indicate that it could be a useful therapeutic tool, where restoration of TIM-3, HBD-2 and the function of Tregs may repair the dysfunctional immunoregulation in CD.

The Expression of Human Beta Defensins by Thermal Injury

PURPOSE: Defensins are important components of innate immune system. These peptides have antimicrobial activity against a wise variety of pathogens that associated with burn wound infection. In particular, human beta-defensins are expressed in normal epidermal region and showed differential expression of some skin disease. We investigated that expression of human beta-defensin by in vitro and ex-vivo by thermal condition. METHODS: To investigate the expression of human beta-defensins in acute burn condition, we cultured keratinocytes and used to rat's skin at this experiment. After thermal condition, we showed the expression of beta-defensins-2 (hBD-2), -3 (hBD-3), keratins, keratinocyte differentiation and junction protein levels by RT-PCR and immunohistochemistry (IHC). RESULTS: HBD-2 & involucrin were down-regulated from 1 hr to 8 hrs in mRNA level. But others were not changed in mRNA level. In protein level, hBD-3 was decreased but pan-cytokeratin and beta-catenin were not changed. CONCLUSION: HBD-2 was down-regulated in thermal injury. Because thermal injury could induce the influence of keratinocyte differentiation and the decrease of skin protection ability. Our results suggested that human beta-defensins plays an important role in protection by several injury.