Objective:To observe the influence of wogonin on colitis mice induced by dextran sulfate sodium (DSS) and explore the related mechanism.Methods:Eighteen C57BL/6 mice were randomly and equally divided into the control group, the model group and the treatment group. The water was given normally to mice in control group, and the 2.5% DSS drinking water was given to mice of other two groups for 7 days. Wogonin via intraperitoneal injection was administrated in the mice of treatment group on the second and the fourth day. The mice were sacrificed on the eighth day and specimens were collected. The pathological damage and inflammation degree of mice colon were evaluated by measuring the length of colon and using HE staining. Immunofluorescence was used to detect the infiltration of neutrophils in mice colon tissue. Wogonin of 25, 50, 100 μmol/L was applied to handle neutrophils from mice marrow tissue in vitro, and there was no treatment in the negative control group. The flow cytometry was used to detect the apoptosis of neutrophil. Western blot was used to detect the expressions of anti-apoptotic protein myeloid cell leukelia-1 (Mcl-1) and extracellular signal-regulated kinase (ERK) . Results:Compared with the model group, the mice colon length in the treatment group was significantly longer [ (7.80 ± 0.21) cm vs. (6.43 ± 0.10) cm, P<0.01], the pathological damage score of the colon tissue was significantly lower [ (6.83 ± 0.98) points vs. (14.33 ± 1.03) points, P<0.01], the number of infiltrative neutrophils in the colon of mice was significantly lower [ (8.52 ± 0.15) neutrophils per low power field vs. (29.43 ± 0.43) neutrophils per low power field, P<0.01]. The apoptosis rate of neutrophils were 6.41% ± 0.51%, 14.01% ± 0.81%, 20.89% ± 0.82%, 24.23% ± 0.29% in negative control group and 25, 50, 100 μmol/L wogonin groups. The apotosis rate of neutrophils increased constantly with the concentration of wogonin increasing gradually and there were significant differences among any two groups ( P<0.01) . Compared with the negative control group, the phosphorylated ERK expressions of neutrophils in 25, 50, 100 μmol/L wogonin groups were decreased obviously (all P<0.05) . The Mcl-1 expression of neutrophils declined constantly with the concentration of wogonin increasing gradually. Conclusion:Wogonin can induce the apoptosis of neutrophils in concentration-dependent manner, reduce the infiltration of neutrophils and relieve the intestinal damage in colon tissue of colitis mice, which may be regulated by the inhibition of ERK phosphorylation and decreased expression of Mcl-1 in concentration-dependent manner.

Objective:To observe the influence of wogonin on colitis mice induced by dextran sulfate sodium (DSS) and explore the related mechanism.Methods:Eighteen C57BL/6 mice were randomly and equally divided into the control group, the model group and the treatment group. The water was given normally to mice in control group, and the 2.5% DSS drinking water was given to mice of other two groups for 7 days. Wogonin via intraperitoneal injection was administrated in the mice of treatment group on the second and the fourth day. The mice were sacrificed on the eighth day and specimens were collected. The pathological damage and inflammation degree of mice colon were evaluated by measuring the length of colon and using HE staining. Immunofluorescence was used to detect the infiltration of neutrophils in mice colon tissue. Wogonin of 25, 50, 100 μmol/L was applied to handle neutrophils from mice marrow tissue in vitro, and there was no treatment in the negative control group. The flow cytometry was used to detect the apoptosis of neutrophil. Western blot was used to detect the expressions of anti-apoptotic protein myeloid cell leukelia-1 (Mcl-1) and extracellular signal-regulated kinase (ERK) . Results:Compared with the model group, the mice colon length in the treatment group was significantly longer [ (7.80 ± 0.21) cm vs. (6.43 ± 0.10) cm, P<0.01], the pathological damage score of the colon tissue was significantly lower [ (6.83 ± 0.98) points vs. (14.33 ± 1.03) points, P<0.01], the number of infiltrative neutrophils in the colon of mice was significantly lower [ (8.52 ± 0.15) neutrophils per low power field vs. (29.43 ± 0.43) neutrophils per low power field, P<0.01]. The apoptosis rate of neutrophils were 6.41% ± 0.51%, 14.01% ± 0.81%, 20.89% ± 0.82%, 24.23% ± 0.29% in negative control group and 25, 50, 100 μmol/L wogonin groups. The apotosis rate of neutrophils increased constantly with the concentration of wogonin increasing gradually and there were significant differences among any two groups ( P<0.01) . Compared with the negative control group, the phosphorylated ERK expressions of neutrophils in 25, 50, 100 μmol/L wogonin groups were decreased obviously (all P<0.05) . The Mcl-1 expression of neutrophils declined constantly with the concentration of wogonin increasing gradually. Conclusion:Wogonin can induce the apoptosis of neutrophils in concentration-dependent manner, reduce the infiltration of neutrophils and relieve the intestinal damage in colon tissue of colitis mice, which may be regulated by the inhibition of ERK phosphorylation and decreased expression of Mcl-1 in concentration-dependent manner.

Objective: To investigate the reverse effect and mechanism of IL-12 on chemotherapeutic medicine suppressing the immune function of NK cells. Methods: Purified NK cells were stimulated with PMAplus Ionomycin in the presence or absent of Cisplatin (DDP) and IL-12. The levels of IFN-γ and TNF-α in culture supernatants were detected by enzyme-linked immunosorbent assay (ELISA); The content of IFN-γ and TNF-α, TRAIL (TNF-related apoptosis inducing ligand) and transcription factors including T-bet and p-STAT-4 in NK cells were analyzed by Flow cytometry. The cytotoxicity of purified NK cells (pretreated with/without chemotherapeutics and IL-12 for 48 h) to Jurkat cells was measured by Flow cytometry. Results: Chemotherapeutics significantly inhibited the production of IFN-γ, TNF-α and the expression of TRAILin NK cells, which were significantly reversed by IL-12 (P<0.05 or P<0.01). Further study revealed that chemotherapeutics down-regulated while IL-12 reversed the expression of p-STAT4 to restore cytokine production. In addition, DDP also inhibited but IL-12 recovered the cytotoxicity of NK cells against tumor cells by inducing the expression of TRAIL (P<0.05 or P<0.01). Conclusion: Chemotherapeutics inhibited the cytotoxicity of NK cells and its secretion of cytokines (IFNγ and TNF-α), which were reversed by IL-12 via up-regulating TRAIL and p-STAT-4; this might provide experimental evidence for the clinical application of IL-12 for rebuild the immune function of tumor patients receiving chemotherapy.

Objective:The occurrence of numerous tumors, particularly classical Hodgkin's lymphoma (CHL), is related with Ep-stein-Barr virus (EBV) infection. However, the incidence of CHL and its association with EBV varies significantly with ethnicity, geo-graphic location, sex, and age. This study investigated the association of EBV infection with CHL in Northern Chinese Han population. Methods:EBV-encoded small RNA (EBER) was detected in 136 cases of CHL through in situ hybridization. Results:A total of 37 cas-es were EBER positive (28%). The mixed cellularity (MC) subtype had the highest positive EBER rate of 49%(23/47;P<0.001), fol-lowed by lymphocyte-rich subtype with 30%(3/10), nodular sclerosis (NS) subtype with 14%(10/73), and 1ymphocyte depletion with 0%(0/2). Our study identified a single age distribution in the third decade. Moreover, NS subtype showed an evident single peak in the third decade. However, MC subtype had a lower peak in the fifth decade. The incidence of EBER showed a bimodal age distribution with two peaks in the first and fifth decades (21.6%and 24.3%, respectively). Conclusion:CHL in Northern Chinese Han population was associated with EBV infection, particularly the MC subtype.