OBJECTIVES: The aim of this study is to determine the effect of cannabinoid receptor (CB) 2 inhibitor on desmoglein 3 (DSG3) expression in HaCaT cells co-cultured with pemphigus serum. METHODS: Immunohistochemical staining was used to compare CB expression in pemphigus patients and normal individuals. Enzyme-linked immunosorbent assay (ELISA) was employed to quantify the concentration of CB2 in the serum of pemphigus patients and normal individuals. A correlation analysis was performed to examine the relationship between the serum CB2 and DSG of pemphigus patients. The CCK-8 assay was used to evaluate the inhibitory effect of AM630 on HaCaT cells, and the half-maximal inhibitory concentration (IC₅₀) value was utilized to determine the experimental concentration. Serum from normal individuals (negative control group) and pemphigus patients (pemphigus group) was co-cultured with HaCaT cells at a 1∶1 ratio. HaCaT cells cultured in complete medium were used as the control group. HaCaT cells in the pemphigus group treated with AM630 were employed as the AM630 group. Real-time polymerase chain reaction (PCR) and Western blot were conducted to assess the expression levels of CB2, DSG3, and β-catenin. Cell dissociation experiments were conducted to evaluate the effect of AM630 on the adhesion of HaCaT cells. RESULTS: Immunohistochemistry revealed significant differences in CB2 expression between pemphigus and normal mucosa (P<0.000 1), but no difference was found in CB1 expression. ELISA analysis revealed a statistically significant difference in the expression levels of CB2 in the serum between normal individuals and pemphigus patients (P<0.001). The expression of CB2 in the serum of pemphigus patients exhibited a significant positive correlation with that of DSG3 (r=0.831, P=0.003). The CCK-8 assay indicated that the IC₅₀ of AM630 on HaCaT cells was 0.55 μmol/L. Real-time PCR and Western blot showed that the expression levels of CB2 and DSG3 increased in the pemphigus group, while the expression level of β-catenin decreased compared with that in the AM630 groups (P<0.05). CONCLUSIONS CB2 is highly expressed in oral mucosal pemphigus. AM630 inhibits overexpression of CB2 and DSG3 and underexpression of β-catenin levels, which can provide new therapeutic targets for pemphigus.
Humans ; Pemphigus/pathology* ; Receptor, Cannabinoid, CB2/metabolism* ; Desmoglein 3/metabolism* ; Acantholysis/metabolism* ; Mouth Mucosa/pathology* ; HaCaT Cells ; Coculture Techniques ; beta Catenin/metabolism*

Objective:To investigate the correlation between venous collateral circulation and clinical data such as symptoms, parenchymal injury, and prognosis in patients with cerebral venous thrombosis(CVT).Methods:The clinical and imaging data of patients with CVT diagnosed in the department of Neurosurgery of the Third Affiliated Hospital of Southern Medical University from December 2011 to August 2018 were retrospectively analyzed. A total of 32 patients with CVT were included, 19 males and 13 females, aged from 20 to 60 (39±12) years. All patients underwent cerebral angiography, individualized delayed rotational phlebography. According to the number and diameter of collateral circulation and the phenomenon of flow delay of contrast medium, the collateral venous circulation scale (CVCS) was developed and divided into 3 levels. The clinical data (risk factors, course of disease, clinical symptoms), imaging data (parenchymal injury, thrombus site), treatment (endovascular treatment, decompressive craniectomy) and prognosis of all patients were recorded. The differences in clinical data, imaging appearances, parenchymal injury, and prognosis between patients with different CVCS were compared, and the correlation between variables with statistically significant differences and CVCS was compared using the Gamma method or Spearman correlation analysis.Results:Among the 32 patients with CVT, 9 were CVCS 0, 13 were CVCS 1 and 10 were CVCS 2. Among them, there were 19 cases of neurological deficit and 17 cases of brain parenchymal injury. There were significant differences in course of disease, neurological deficit, focal dyskinesia, language dysfunction, consciousness disorder, isolated headache, deep vein thrombosis, cortical vein thrombosis and prognosis across different CVCS ( P<0.05). Correlation analysis showed that CVCS was positively correlated with course of disease and isolated headache ( r=0.724, 0.637, P<0.001), and negatively correlated with neurological deficit symptoms, focal dyskinesia, disturbance of consciousness, brain parenchymal injury and deep vein thrombosis ( r=-0.797, -0.451, -0.782, -0.697, -0.427, P<0.05). The results of 90 days follow-up showed that there were 18 cases with mRS 0, 6 cases with mRS 1, 2 cases with mRS 2-4, and 1 case with mRS 5-6 points. There was a negative correlation between CVCs grading and mRS score at 90 days ( r=-0.732, P<0.001). Conclusion:Lower cerebral venous collateral circulation grade is associated with higher incidence of brain parenchymal injury, neurological deficit symptoms, and worse clinical prognosis.