AIM:To observe the expression of TLR2 and TLR4 on mast cells (MCs) in the periapical tissues from different types of human chronic periapical diseases , and to analyze the role of TLR 2 and TLR4 on tryptase-positive MCs in the immunopathogenesis of human chronic periapical diseases .METHODS: A total of 60 donors, including healthy control group , periapical granuloma group and periapical cyst group , were enrolled in the study .The periapical tis-sue specimens were fixed in 10%buffered formalin and stained with hematoxylin and eosin for histopathology , or stained with double-immunofluorescence for identification of TLR 2-tryptase and TLR4-tryptase double-positive MCs in the periapical tissues.RESULTS:Compared with the healthy control , the densities of TLR2-tryptase and TLR4-tryptase double-positive MCs in periapical tissues were significantly increased in human chronic periapical diseases (P<0.01).The densities of TLR2-tryptase and TLR4-tryptase double-positive MCs in periapical cyst group were significantly higher than those in peria-pical granuloma group (P<0.01).CONCLUSION:TLR2 and TLR4 were expressed on the MCs in the periapical tissues of human chronic periapical diseases .TLR2-tryptase and TLR4-tryptase double-positive MCs may participate in the patho-genesis of chronic periapical diseases .

Background Extracellular matrix (ECM) fibrosis leads to corneal scaring during the process of cornea wound healing.Transforming growth factor-β1 (TGF-β1) is known to mediate overproduce of ECM components.Our previous study developed a three-dimensional model for corneal stromal cells culture in vitro.Objective The hypothesis of this study was to apply TGF-β1 in the three-dimensional culture system to establish a corneal stroma ECM fibrosis model.Methods Fresh bovine corneas were extracted for the culture of bovine keratocytes in constructed three-dimension culture system.The Pellets were cultured in the DMEM/F12+ 10％ fetal bovine serum (FBS) medium with 0.5 ng/ml or 1.0 ng/ml TGF-β1 or without TGF-β1,respectively.Calcein AM/(propidium iodide) PI staining was employed to assay the cell viability 2 weeks after culture.The expressions of α-smooth muscle actin (α-SMA),type Ⅰ collagen (Col Ⅰ) and Col Ⅲ mRNA and protein in the cells were detected by real-time PCR and Western blot respectively 48 hours,1 week and 2 weeks after cultured.The results were statistically analyzed.Results Cultured for 48 hours in the Pellet system,corneal stromal cells clustered and was identified alive by Calcein-AM/PI staining in 2 weeks.The relative expression levels of α-SMA,Col Ⅰ and Col m mRNA were elevated in both the 0.5 ng/ml and 1.0 ng/ml TGF-β1 supplement groups in comparison with the only DMEM/F12+10％ FBS group,with marked difference among the three groups (Fgroup =696.745,P＜0.001;Fgroup =35.166,P＜0.001;Fgroup =33.677,P＜0.001),and the expression levels increased with the lapse of culture time (Ftime =5.863,P＜0.05;Ftime =298.614,P＜0.001;Ftime =607.472,P＜0.001).The synthetic rate of Col Ⅲ mRNA was obviously faster than that of Col Ⅰ mRNA.Western blot showed that only a trace of α-SMA,Col Ⅰ and Col Ⅲ were detected 48 hours and 1 week after culture.The expression levels of α-SMA,Col Ⅰ and Col Ⅲ in Pellet system in 0.5 ng/ml TGF-β1 medium were 0.395±0.208,1.060±0.175 and 0.629±0.382,and in 1.0 ng/ml TGF-β1 medium were 0.758±0.228,1.201 ±0.187 and 0.753±0.468,respectively 2 weeks after culture,significant differences were shown among the three groups (α-SMA:F=10.691,P＜0.05;Col Ⅰ:F=14.094,P＜0.05;Col Ⅲ:F=10.995,P＜0.05).Conclusions Addition of TGF-β1 and serum enhance the assembly and fibrosis of ECM,showing the higher expressions of specific fibrotic markers in bovine keratocytes Pellet.This culture systerm can be used as a candidate three-dimensional model for corneal stroma ECM fibrosis.

Myeloid differentiation factor 88 (MyD88) plays an important role in tumorigenesis,development and malignant transformation,also participates in the microenvironment,proliferation,apoptosis,invasion,metastasis and tumor resistance.MyD88 may serve as a new and meaningful therapeutic target,which can promote the growth and development of tumors by regulating multiple signaling pathways and enhance the drug resistance of tumor cells.

Objective:To determine the prevalence situation of adult female acne (AFA) patients and to explore the risk factors related to AFA.Methods:From September 2019 to June 2020, 290 female acne patients aged from 25 to 48 (29.57±4.50) years were surveyed with a questionnaire of risk factors and the prevalence situation of acne in the acne clinic of the Department of Dermatology, Wuxi No.2 Hospital Affiliated with Nanjing Medical University.Results:AFA occured more frequently in the jaw (95.17%), cheek (93.79%) and forehead (89.66%). Recurrent acne (38.62%) and comedonal acne (50.34%) more commonly occured. Cosmetics, endocrine, diet, genetics and other factors aggravated AFA. Age ( H=7.286, P>0.05; F=0.122, P>0.05), gonadal hormone concentrations ( Z=-0.365, P>0.05; χ 2=0.276, P>0.05), menstrual cycle ( Z=-0.274, P>0.05; χ 2=0.217, P>0.05), genetics ( Z=-1.244, P>0.05; χ 2=1.771, P>0.05) made no difference to acne grading and types. Excessive use of cosmetics could lead to increased comedo (χ 2=7.097, P<0.05). Cosmetics had no difference to acne grading ( Z=-0.065, P>0.05). Gonadal hormone concentrations were uncorrelated with menstrual cycle (χ 2=1.397, P>0.05). Conclusions:The pathogenesis of AFA is related to a variety of factors, which affect the skin barrier function and require comprehensive treatment.