Forty adult male C57BL/6 mice were randomly divided into four groups:the control group,Salmonella typhimurium(S.typhimurium)(ST)group,phlorizin(PHZ)+S.typhimuri-um(ST)group,and PHZ(80 mg/kg)group,with 10 mice in each group.Morphological observa-tion,HE staining,ELISA,immunofluorescence and Western blot were performed,the results showed that PHZ significantly increased the cecal index,decreased the spleen index of S.typhi-murium-induced mice(P＜0.05),and reduce the pathological damage of cecum in mice.Mean-while,PHZ treatment also significantly reduced colonization of S.typhimurium in the cecum,spleen,mesenteric lymph nodes and liver(P＜0.05).The results of ELISA showed that PHZ treatment also significantly inhibited the S.ty phimurium-induced increase in the expression of IL-1β,INF-γ,TNF-a and IL-6 in the cecum of mice(P＜0.05).Immunofluorescence and Western blot results showed that PHZ significantly increased the protein expression levels of Occludin,Claudin-3,and ZO-1 in the cecal barrier of mice induced by S.typhimurium(P＜0.05).These results con-firmed that phlorizin could improve cecal inflammation and intestinal barrier damage induced by S.typhimurium in mice.

Mastitis is one of the common and prevalent diseases in dairy cows,and the natural prod-uct daidzein is a kind of natural flavonoids with a wide range of pharmacological and anti-inflam-matory effects.However,the effect of daidzein on mastitis in dairy cows has not been reported.Therefore,in this study,we explored the effects of daidzein on LPS-induced inflammatory response and tight junction damage in dairy mammary epithelial cells Firstly,we pretreated the MAC-T cell line using different concentrations of daidzein,and it was clarified that daidzein below 200 μmol/L had no effect on the cell activity.Next,we examined the production of pro-inflammatory mediators in LPS-stimulated MAC-T cell lines using qRT-PCR,and clarified that daidzein could reduce the production of pro-inflammatory mediators in a concentration-dependent manner.Subsequently,af-ter the expression of Occludin,Claudin3 and ZO-1 was detected by immunofluorescence and West-ern Blot,it was clear that daidzein could alleviate MAC-T cell intercellular tight junction injury.Fi-nally,it was demonstrated that daidzein significantly inhibited LPS-induced activation of the NF-κB signaling pathway within MAC-T using network pharmacological analysis and Western Blot.The above results suggest that daidzein can inhibit LPS-induced inflammatory response and tight junc-tion damage in mammary epithelial cells of cows by suppressing the activation of NF-κB signaling pathway.The present study provides a theoretical basis for the alleviation of mastitis by natural products and further expands the pharmacological effects of daidzein.

Mastitis is one of the common and prevalent diseases in dairy cows,and the natural prod-uct daidzein is a kind of natural flavonoids with a wide range of pharmacological and anti-inflam-matory effects.However,the effect of daidzein on mastitis in dairy cows has not been reported.Therefore,in this study,we explored the effects of daidzein on LPS-induced inflammatory response and tight junction damage in dairy mammary epithelial cells Firstly,we pretreated the MAC-T cell line using different concentrations of daidzein,and it was clarified that daidzein below 200 μmol/L had no effect on the cell activity.Next,we examined the production of pro-inflammatory mediators in LPS-stimulated MAC-T cell lines using qRT-PCR,and clarified that daidzein could reduce the production of pro-inflammatory mediators in a concentration-dependent manner.Subsequently,af-ter the expression of Occludin,Claudin3 and ZO-1 was detected by immunofluorescence and West-ern Blot,it was clear that daidzein could alleviate MAC-T cell intercellular tight junction injury.Fi-nally,it was demonstrated that daidzein significantly inhibited LPS-induced activation of the NF-κB signaling pathway within MAC-T using network pharmacological analysis and Western Blot.The above results suggest that daidzein can inhibit LPS-induced inflammatory response and tight junc-tion damage in mammary epithelial cells of cows by suppressing the activation of NF-κB signaling pathway.The present study provides a theoretical basis for the alleviation of mastitis by natural products and further expands the pharmacological effects of daidzein.

Forty adult male C57BL/6 mice were randomly divided into four groups:the control group,Salmonella typhimurium(S.typhimurium)(ST)group,phlorizin(PHZ)+S.typhimuri-um(ST)group,and PHZ(80 mg/kg)group,with 10 mice in each group.Morphological observa-tion,HE staining,ELISA,immunofluorescence and Western blot were performed,the results showed that PHZ significantly increased the cecal index,decreased the spleen index of S.typhi-murium-induced mice(P＜0.05),and reduce the pathological damage of cecum in mice.Mean-while,PHZ treatment also significantly reduced colonization of S.typhimurium in the cecum,spleen,mesenteric lymph nodes and liver(P＜0.05).The results of ELISA showed that PHZ treatment also significantly inhibited the S.ty phimurium-induced increase in the expression of IL-1β,INF-γ,TNF-a and IL-6 in the cecum of mice(P＜0.05).Immunofluorescence and Western blot results showed that PHZ significantly increased the protein expression levels of Occludin,Claudin-3,and ZO-1 in the cecal barrier of mice induced by S.typhimurium(P＜0.05).These results con-firmed that phlorizin could improve cecal inflammation and intestinal barrier damage induced by S.typhimurium in mice.

Objective: To analyze the clinical characteristics and treatment experience of Vitamin B12 (VB12) deficient patients with recurrent aphthous ulcers (RAU) to improve the clinical efficacy. Methods: A retrospective analysis was performed on 15 cases of recurrent oral ulcers from January 2016 to September 2018. The causes were analyzed according to the patients’ clinical characteristics. Results: In total, 15 patients with RAU had no remission after routine immunotherapy. Further clinical examination suggested that vitamin B12 levels were reduced. The erythrocyte mean corpuscular volume (MCV) was significantly increased, and the average number of red blood cells (RBC) and hemoglobin (Hb) levels were decreased. RAU disappeared after vitamin B12 supplementation. Routine blood work showed that the MCV returned to the normal range, which was statistically significant compared with the pretreatment MCV (P < 0.001). Vitamin B12 serum levels were significantly higher (P < 0.001) than those before treatment. Conclusion When the main manifestation of vitamin B12 deficiency is recurrent oral ulcer symptoms, dentists should examine the lesions carefully, inquire about the medical history in detail, and perform further serological tests when necessary to avoid the overuse of immunosuppressive drugs for treatment.

Objective: To detect the expression of vascular endothelial growth factor (VEGF), intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) in oral lichen planus (OLP). Methods : Thirty OLP patients and fifteen healthy subjects were enrolled in the study. Serum were collected from 15 healthy volunteers as controls. Normal tissues were collected from surgical department as immunohistochemical analysis. The levels of VEGF, ICAM-1, VCAM-1 in serum were measured by ELISA. Immunohistochemical analysis of VEGF, ICAM-1, VCAM-1 were carried out by the means of primary antibodies and anti-VEGF, anti-CD106 antigen (VCAM-1) and anti-CD54 antigen (ICAM-1). Results: ELISA results showed no expression differences for VEGF between the two groups. Whereas, the levels of ICAM-1, VCAM-1 in OLP group were significantly higher than those in control group (P < 0.05). Immunohistochemical results reveal the presence of a significant angiogenesis in OLP patients through the immunoexpression of VEGF, ICAM-1, VCAM-1 according to the percentage of stained cells (P < 0.05). Conclusion Regarding the results, it seems that high expression of VCAM-1 and ICAM-1 are related to oral lichen planus.