Objective To investigate the effect of propofol on the function of gap junction (GJ) in HeLa cells transfected with Cx32/Cx26 plasmid. Methods Cervical cancer HeLa cells transfected with Cx32/Cx26 was given as present by professor Andrew L. Harris from New Jersey Dental Medical School, department of pharmacology and physiology. The transfected cells were selected by G-418. The effective GJ channels were identified by "Parachute Assay". The cells were randomly divided into 6 groups: Ⅰ control group (group C); Ⅱ fat emulsion group was exposed to fat emulsion 10 μg/ml (group E); Ⅲ 18-α-GA group was exposed 18-α-GA (gap junction blocker) 1.0 μg/ml (18-α-GA); Ⅳ, Ⅴ, Ⅵ propofol groups were exposed to propofol 1.3, 2.2 and 3.2μg/ml respectively (group P1, P2, P3). The transfected HeLa cells were incubated at 37 ℃ for 4 h. Gap junction function was assessed using fluorescent indicators Calcine-AM which emits green fluorescence and CM-Dil which emits red fluorescence. The small molecular Calcine-AM can pass through gap junction and enters HeLa cells while the large molecular CM-Dil cannot pass through gap junction and stays in the loading cells. Fluorescent indicator transmissibility and inhibition rate were calculated. Results The fluorescent indicator transmissibility was significantly lower and inhibition rate higher in group 18-α-GA, P1, P2 and P3 than in control group. There was nosignificant difference in the fluorescent indicator transmissibility and inhibition rate between group C and E. The inhibition of GJ function by propofol was dose-dependent. Conclusion Propofol can inhibit the function of GJ in HeLa cells transfected by Cx32/Cx26 in a dose-dependent manner.

Aim To study the effect of emodin in Po-lygonum multiflorum on the expression of CYP450 isoenzymes in L02 hepatocytes and explore its mecha-nism of cytotoxicity. Methods L02 cells were treated with different concentrations of emodin. Cell viability was examined by MTS assay kit, and cell membrane injury was examined by detecting the release rate of lactate dehydrogenase( LDH) . The expression of cyto-chrome P450 mRNA was detected by real time PCR. Results The result of MTS assay showed that L02 cells viability was significantly reduced following expo-sure to emodin in a concentration and time dependent manner. The LDH release rate of L02 cells significant-ly increased after exposure to emodin for 48 h com-pared with the control group. On the mRNA level, compared with the control group,emodin had inductive effects on mRNA of each CYP450 enzyme, while had significant inductive effects on mRNA of CYP1 A1 and CYP1 B1 in a concentration and time dependent man-ner. Conclusion Emodin in Polygonum multiflorum may generate significant liver injury in L02 cells and has inductive effects on CYP450 enzyme activity.

Objective To investigate the protective effect of wogonin on acute kidney injury(AKI)induced by li-popolysaccharide(LPS).Methods The model of septic-induced AKI was established on male C57BL/6J mice by a single intraperitoneal injection of LPS and normal C57BL/6J mice were used as normal control group.Twenty-four male C57BL/6 mice were randomly divided into 4 groups(6 mice in each group):normal control group(NC),normal control+wogonin(NC+WOG 12.5 mg/kg),LPS model group(LPS 10 mg/kg),LPS model+wogonin(LPS 10 mg/kg+WOG 12.5 mg/kg).After LPS intervention for 24 h,serum samples were collected to detect blood creatinine(CRE)and urea nitrogen(BUN)levels.HE staining and PAS staining were performed to observe the degree of renal pathological injury.Immunohistochemistry was performed to detect the degree of expression of inflammatory markers interleukin(IL)-1 β,IL-6 and tumor necrosis factor(TNF)-α in renal tissues.PCR was performed to detect the expression of KIM-1 and NGAL in renal tissues.Western blot was performed to detect the changes in protein expression of NF-κB signaling pathway subunits P65 and PP65 in renal tissues.Results Com-pared with NC group,CRE and BUN levels in LPS group increased(FCRE=60.90,P＜0.001,FBUN=82.13,P＜0.001);compared with LPS group,these indexes decreased in LPS+WOG group(P＜0.001).PCR test re-sults showed that compared with the NC group,the expression of KIM-1 and NGAL mRNA was significantly in-creased in LPS group(FKIM-1=146.3,P＜0.001,FNGAL=161.2,P＜0.001).In contrast,KIM-1 and NGAL mR-NA expression was decreased in the LPS+WOG group(P＜0.01).Renal histopathological examination showed that compared with the NC group,renal tissues of mice had renal tubular dilatation and inflammatory cell infiltration in LPS group;compared with LPS group,the number of tubular dilatation reduced and inflammatory cell infiltration was reduced in LPS+WOG group(FHE=721.4,P＜0.001;FPAS=518.9,P＜0.001).Immunohistochemical staining showed that the expression of IL-1 β,IL-6 and TNF-α was significantly increased in the LPS group;com-pared with the LPS groups(FIL-1β=114.6,FIL-6=108.9,FTNF α=251.6,all P＜0.001),these indexes de-creased in LPS+WOG group(all P＜0.01).Further studies using Western blot showed that the NF-κB signaling pathway of LPS-treated mice had been activated and produced a hyperphosphorylated state in comparison to the NC group(FPP65=13.02,P＜0.01),yet this pathway in the LPS+WOG group showed the opposite effect,namely attenuated activity and reduced phosphorylation when the control was LPS(P＜0.01).Conclusion WOG effec-tively blocked the NF-κB signaling pathway in the LPS-induced acute kidney injury model mice,thereby attenua-ting the inflammatory response and tissue damage in the kidneys of LPS-induced acute kidney injury mice.

Objective To analyze the clinical efficacy of different anti-tumor therapies for recurrence and metastasis after liver transplantation for primary liver cancer (liver cancer). Methods Clinical data of 145 recipients undergoing liver transplantation for liver cancer were retrospectively analyzed. The overall survival and recurrence and metastasis after liver transplantation for liver cancer were analyzed. The clinical efficacy of different anti-tumor therapies for recipients with recurrence and metastasis were compared. Results Sixty-five recipients (44.8%) developed recurrence and metastasis. The median recurrence time was 6 months. Among them, 1 case underwent secondary liver transplantation after recurrence and died of intestinal perforation. Twenty-four recipients (37%) received targeted drug therapy with a median tumor-bearing survival of 22 months. Eleven recipients (17%) received radiotherapy or chemotherapy with a median tumor-bearing survival of 11 months. Nine recipients (14%) received local treatment (surgical resection or radiofrequency ablation), and the median tumor-bearing survival was 8 months. Twenty recipients (31%) abandoned anti-tumor therapy, and the median tumor-bearing survival was 3 months. The tumor-bearing survival of recipients receiving anti-tumor therapy was significantly longer than that of recipients without anti-tumor therapy (P < 0.001). The tumor-bearing survival of recipients receiving targeted drug therapy was significantly longer than that of those receiving other anti-tumor therapies (P=0.03). The tumor-bearing survival of recipients receiving local treatment, radiotherapy and chemotherapy was considerably longer than that of those who abandoned anti-tumor therapy (P=0.004). Conclusions Surgical resection and radiofrequency ablation are the optimal therapies for recipients with recurrence and metastasis after liver transplantation for liver cancer. For recipients with multi-focal tumors who fail to receive local treatment, those receiving targeted drug therapy obtain the longest survival. In addition, radiotherapy and chemotherapy can also prolong the survival of recipients with recurrence and metastasis.

Objective:To explore the efficacy of sirolimus-based immunosuppressive protocol on tumor recurrence and tumor-free survival after liver transplantation(LT)in hepatocellular carcinoma (HCC)patients.Methods:From January 1, 2016 to December 31, 2018, a total of 114 HCC patients undergoing LT were recruited and divided into two groups of sirolimus(SRL)and tacrolimus. Univariate and multivariant analyses were performed for evaluating the risk factors of recurrence after LT. Tumor-free survival were compared using Cox logistic regression analysis.Results:Tumor recurrence and/or metastasis occurred in 45 patients. Univariate and multivariate regression analysis indicated that sirolimus was an independent protective factor for preventing tumor recurrence( P=0.005, HR=0.38, 95% CI 0.193~0.748). The median tumor-free survival time was 5(4~19)months in tacrolimus group and 23(13~31)months in sirolimus group. No inter-group statistical difference existed in incidence of infection or rejection complications( P>0.05). Conclusions:HCC patients benefit from sirolimus-based immunosuppressive protocol after LT. And sirolimus may reduce tumor recurrence rate and prolong tumor-free survival time.