Objective:To investigate the effects of fluoride exposure on proliferation, apoptosis and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in mice.Methods:BMSCs were isolated and cultured from femur bone marrow of C57BL/6 mice (6 - 8 weeks). The cells in passage 3 were used to detect the surface markers of stem cells by flow cytometry. The cells were cultured in media with a final fluoride concentration of 0.0, 0.1, 1.0, 5.0, 10.0, 15.0, 20.0 and 40.0 mg/L, respectively. The effects of different fluoride concentrations on BMSCs cell proliferation (CCK8 method), apoptosis (flow cytometry analysis), osteogenic differentiation ability [alizarin red and alkaline phosphatase (ALP) staining] were detected. Western blot was applied to detect the levels of apoptosis-related proteins [poly ADP-ribose polymerase (PARP)], mitogen-activated protein kinase (MAPK) pathway member proteins [extracellular regulated protein kinase 1/2 (ERK1/2), c-Jun amino-terminal kinase (JNK), p38 and phosphorylated ERK, JNK, p38 (p-ERK, p-JNK, p-p38)], osteogenic differentiation-related protein [Runt-related transcription factor 2 (Runx2), ALP] and Wnt/β-catenin pathway member proteins [glycogen synthase kinase-3β (GSK3β), phosphorylated GSK3β (p-GSK3β) and β-catenin]. Immunocytofluorescense staining was applied to evaluate the expression levels of p-GSK3β and β-catenin. The two pathways (MAPK and Wnt/β-catenin) were blocked by SP600125 and DKK-1, respectively, to testify their involvement in mechanisms of apoptosis and osteogenic differentiation.Results:The mouse BMSCs were successfully isolated and cultured. Flow cytometry analysis showed that the mesenchymal stem cell surface biomarkers (CD73, CD90 and CD105) were positively expressed. The comparison of cell proliferation at three time points (24, 48 and 72 h) in each concentration group was statistically significant ( F = 65.36, 160.04 and 365.32, P < 0.001), and the comparison of early apoptosis (24 h) in each concentration group was statistically significant ( F = 214.04, P < 0.001); compared with the 0.0 mg/L group, the cell proliferation in 15.0, 20.0 and 40.0 mg/L groups decreased, and the early apoptosis rate in 10.0, 15.0 and 20.0 mg/L groups increased ( P < 0.05). When cells were treated with 15.0 mg/L fluoride for 0 - 24 h, the p-JNK/JNK ratio was higher at 2, 4, 8, 12, 18 and 24 h compared with that at 0 min ( P < 0.05); compared with the fluoride group (15.0 mg/L), the early apoptosis rate of cells after SP600125 block decreased ( P < 0.05), and the protein expression levels of PARP and p-JNK decreased ( P < 0.05). After osteogenic induction, compared with the 0.0 mg/L group, in 0.1 and 1.0 mg/L groups ALP staining was enhanced and the number of calcified nodules increased, and the protein expression levels of Runx2 and ALP in the 0.1 and 1.0 mg/L groups were higher ( P < 0.05). After osteogenic induction, compared with the 0.0 mg/L group, the p-GSK3β/GSK3β ratio and β-catenin protein level were significantly higher in the 0.1 and 1.0 mg/L groups ( P < 0.05); and compared with the fluoride group (1.0 mg/L), addition of DKK-1 significantly decreased the protein expression levels of p-GSK3β and β-catenin and reduced the nuclear entry of β-catenin, and ALP staining decreased and the number of calcified nodules decreased. Conclusions:High concentration of fluoride (> 10.0 mg/L) inhibits the proliferation and promotes apoptosis of BMSCs, while low concentration of fluoride (0.1, 1.0 mg/L) promotes osteogenic differentiation. The MAPK/JNK pathway and the classical Wnt pathway are involved in the above cellular processes, respectively.

Objective:To explore the feasibility and efficacy of one layer interrupted strengthening suture applied in cervical mechanical anastomosis of esophageal cancer operation .Methods:Forty patients with upper and middle esophageal carcinoma , who underwent cervical mechanical anastomosis during operation ,were divided into two groups .In group A ,22 patients re‐ceived no further treatment for the anastomotic stoma after anastomosis .In group B ,18 patients received one‐layer interrupted strengthening suture with 3‐0 prolene for the anastomotic stoma after mechanical anastomosis .Results:The cervical mechanical anastomosis reached one‐time success in all 40 patients .In group A ,4 cases of anastomotic leakage and 3 cases of anastomotic stricture occurred after operation and were cured .No anastomotic leakage and anastomotic stricture was identified in group B . Conclusions:The application of anastomotic one layer interrupted strengthening suture in cervical mechanical anastomosis of esophageal cancer operation could significantly reduce the incidence rates of anastomotic leakage and stricture .It is worthy of promotion and application .

Purpose To investigate the clinical value of artificial intelligence(AI)quantitative parameters in predicting the invasion degree of lung adenocarcinoma with diameter≤2 cm of ground-glass density.Materials and Methods A total of 80 patients with lung adenocarcinoma with diameter≤2 cm ground-glass density confirmed by pathology from March 2019 to April 2022 were retrospectively analyzed.A total of 90 nodules were rerolled,including 8 adenocarcinomas in situ(AIS),34 minimally invasive adenocarcinomas(MIA)and 48 invasive adenocarcinomas(IAC).They were divided into the experimental group(IAC)and the control group(AIS and MIA).The differences of the AI quantitative parameters such as volume,three-dimensional length diameter,maximum area,maximum CT value,minimum CT value and average CT value were compared between two groups,and the predictive values of AI quantitative parameters for the invasion degree of lung adenocarcinoma was evaluated.Results There were statistically significant differences with age,volume,three-dimensional length diameter,maximum area,maximum CT value and average CT value between the two groups(all P<0.05),but no statistically significant differences in gender and minimum CT value(both P>0.05).Binary Logistic regression analysis showed that the three-dimensional length diameter(odd ratio=2.020,P=0.034)and the maximum CT value(odd ratio=1.008,P=0.013)were independent predictors for lung adenocarcinoma with diameter≤2 cm of ground-glass density.The regression model based on the three-dimensional length diameter and the maximum CT value had the best predictive performance,and its AUC was 0.901.When the critical value was 2.432,its sensitivity and specificity were 93.75%and 71.43%,respectively.Conclusion AI quantitative parameters have a high value in predicting the degree of invasion of lung adenocarcinoma with diameter≤2 cm of ground-glass density,and the combined model with three dimensional long diameter and maximum CT value has the highest diagnostic efficiency.