Objective To investigate the impact of shikonin(SHI)on the malignant biological activity of liver cancer cells by regulating Notch signaling pathway.Methods Western blot assay was used to detect the expression of Notch,Hairy mitosis-related enhancer-1(Hes1),hairy-related transcription factor-1(HEY1)protein in liver cancer tissue,paracancerous tissue,hepatoma cells(HepG2 cells,Hep3B cells,HCCLM3 cells,Huh-7 cells and SMMC-7721 cells)and normal liver cells(HL-7702 cells).Huh-7 cells were divided into the control group,the L-SHI group(1μmol/L SHI),the M-SHI group(2μmol/L SHI),the H-SHI group(4μmol/L SHI),the DAPT group(50μmol/L Notch signal inhibitor DAPT)and the H-SHI+VPA group[4μmol/L SHI and 8 mmol/L Notch pathway activator Valproic acid(VPA)].The proliferation of Huh-7 cells was detected by CCK-8 method and plate cloning test.The apoptosis and cell cycle of Huh-7 cells were detected by flow cytometry.Cell scratch test and Transwell invasion test were used to detect migration and invasion of Huh-7 cells.Western blot assay was used to detect the expression of epithelial-mesenchymal transformation(EMT)and apoptosis related proteins.Results The expression levels of Notch,HES1 and HEY1 were obviously increased in liver cancer tissue and cells,and Huh-7 cells showed the most obvious difference,therefore,Huh-7 cells were taken as the research object.Compared with the control group,the protein levels of Notch,HES1,HEY1 and Bcl-2 decreased,and the proportions of S phase and G2 phase cells,OD450 value,number of clones,migration rate,number of invasive cells and levels of N-cadherin and Vimentin decreased significantly in the L-SHI group,the M-SHI group,the H-SHI group and the DAPT group(P＜0.05).The proportion of G1/G0 phase cells,apoptosis rate and levels of Bax,cleaved Casase-3,and E-cadherin increased obviously(P＜0.05).The effect of SHI was dose-dependent.Compared with the H-SHI group,the above indexes showed the opposite trend in the H-SHI+VPA group.VPA attenuated the effect of SHI on reducing the malignant biological activity of liver cancer cells.Conclusion SHI may inhibit the proliferation,migration and invasion of Huh-7 cells and promote apoptosis of Huh-7 cells by inhibiting Notch signal pathway.

【Objective】 To investigate the prognosis of blood donors with occult hepatitis B virus infection (OBI) by long-term follow-up and repeated testing of HBsAg and HBV DNA. 【Methods】 From January 1, 2010 to December 31, 2020, voluntary blood donors were screened by both serological and viral nucleic acid(NAT) testing, then samples were further confirmed as HBV DNA positive by manual nested-PCR amplification.A total of 306 cases were detected as HBsAg negative /HBV DNA positive, then followed-up for a long time and re-examined of HBsAg and HBV DNA to confirm whether they had infected with OBI.The prognosis of patients with OBI who experienced long-term immunization was determined by repeated testing. 【Results】 A total of 306 HBsAg negative/ HBV DNA positive blood donors had been followed up, and 40(13.07%, 40/306) were recalled frequently for re-examination.Among them, 90%(36/40), 57.5%(23/40), 40% (16/40)were anti-HBc + , anti-HBs + and anti-HBe + , respectively, and 50%(20/40), 40%(16/40), 7.5%(3/40) and 2.5% (1/40)were anti-HBs+ / anti-HBc + , anti-HBc + / anti-HBs -, anti-HBc -/ anti-HBs + and anti-HBc -/ anti-HBs -, respectively.Those 40 blood donors were followed-up for 1-13 times, with the duration of 8-108 months (0.6~9 years).1 donor (2.5%) was followed-up less than 1 year, 11 (27.5%)>1 year and ≤3 years, 23 (57.5%) 23(57.5%)>3 years and ≤5 years, and 5 (12.5%) for more than 5 years.After long-term following up and repeated testing, 50%(20/40)of OBI blood donors turned negative for HBV DNA (HBsAg negative / HBV DNA negative), 42.5% (17/40)were confirmed as OBI infection (HBsAg negative / HBV DNA positive), and 7.5%(3/40) were hard to determine (after repeated testing, the results were either positive or negative). 【Conclusion】 After long-term following up and repeated screening, we found that none of the OBI patients turned into acute or chronic HBV infection, and most of them maintained OBI.However, OBI blood donors carry very low load of HBV DNA for a long time, which could lead to false negative results of NAT and bring a great challenge to the safety of blood transfusion.