Objective To investigate the effects of Carthamus tinctorius L.extract(CTLE)on the levels of oxidative stress and inflammation in mice with ethanol-induced alcoholic liver disease and its mechanism of action.Methods SPF-grade C57BL/6 male mice were randomly divided into four groups:control group,model group,low-CTLE group(50 mg·kg-1),and high-CTLE group(100 mg·kg-1).The control group was given Lieber-Decarli liquid diet,and the other groups were given Lieber-Decarli alcohol diet to construct a chronic alcoholic liver injury model in mice.Serum and liver tissues of mice were collected and serum biochemical indexes of mice were detected.HE and oil red O staining were applied to observe pathological changes in mouse liver tissues.Real-time fluorescence quantitative PCR and Western Blot were used to detect the mRNA and protein expression levels of Keap1/Nrf2 and STAT3/NF-κB pathway-related factors.Results Compared with the model group,the ALT,AST,LDL-C,and MDA levels were significantly reduced(P＜0.05,P＜0.01),while the levels of HDL-C,SOD,and GSH were increased dramatically in the administered group(P＜0.05,P＜0.01),which indicated that CTLE has specific protective and antioxidant effects on alcoholic liver injury in mice.HE staining and oil red O staining showed that the hepatic lesions and lipid deposition of mice were ameliorated.It enhances the antioxidant and anti-inflammatory effects of the body by activating the mRNA and protein expression levels of antioxidant factors related to the Keap1/Nrf2 pathway and inhibiting the mRNA and protein expression levels of inflammatory factors related to STAT3/NF-κB pathway(P＜0.05,P＜0.01).Conclusion It was shown that CTLE could exert anti-oxidative stress and anti-inflammatory effects through regulating Keap1/Nrf2 and STAT3/NF-κB signaling pathways to attenuate alcoholic liver injury in mice.This study may provide a new idea for the treatment of alcoholic liver disease and the subsequent study of molecular mechanisms.

Abstract OBJECTIVE To study the effect of berberine(BBR) on autophagy of human breast cancer HCC1937 cells under hypoxia condition. METHODS Cultured human breast cancer HCC1937 cells, CCK-8 method was used to determine the effects of different concentrations of BBR(0, 5, 10, 20, 40, 80, 160 μmol·L-1) on cell viability under normoxia and hypoxia conditions, and select the drug concentration for further experiments. Cultured HCC1937 cells were randomly divided into 4 groups: control group, 20 μmol·L-1 BBR group, hypoxia group, hypoxia+20 μmol·L-1 BBR group. LIVE/DEAD cell viability/cytotoxicity kits were used to measure the cell death rate. The expressions of autophagy related proteins Beclin1, LC3 and P62 in each group were determined by Western blotting. The cells were infected with mCherry-GFP-LC3 adenovirus, and the number of autophagosomes and autophagolysosomes in each group were counted by laser confocal microscopy to determine the effect of BBR on the autophagy flow of HCC1937 cells. RESULTS BBR decreased the cell viability of human breast cancer HCC1937 cells in a concentration-dependent manner. After hypoxia treatment, the cell death rate of HCC1937 cells was not significantly changed, and the intracellular Beclin1, LC3-II and LC3-II/LC3-I ratio were significantly increased, while P62 without significant changes, and the autophagy flow was increased. BBR significantly increased cell death rate, decreased Beclin1 and LC3II/LC3-I ratio, increased intracellular P62, significantly reduced the number of autophagosomes and autophagolysosomes, and inhibited the formation and clearance of autophagosomes under both normal and hypoxia conditions. CONCLUSION BBR increases the death rate of human breast cancer HCC1937 cells under hypoxia condition, and its effect is related to the inhibitory effect of berberine on autophagy under hypoxia condition.

New threats posed by the emerging circulating variants of SARS-CoV-2 highlight the need to find conserved neutralizing epitopes for therapeutic antibodies and efficient vaccine design. Here, we identified a receptor-binding domain (RBD)-binding antibody, XG014, which potently neutralizes β-coronavirus lineage B (β-CoV-B), including SARS-CoV-2, its circulating variants, SARS-CoV and bat SARSr-CoV WIV1. Interestingly, antibody family members competing with XG014 binding show reduced levels of cross-reactivity and induce antibody-dependent SARS-CoV-2 spike (S) protein-mediated cell-cell fusion, suggesting a unique mode of recognition by XG014. Structural analyses reveal that XG014 recognizes a conserved epitope outside the ACE2 binding site and completely locks RBD in the non-functional "down" conformation, while its family member XG005 directly competes with ACE2 binding and position the RBD "up". Single administration of XG014 is effective in protection against and therapy of SARS-CoV-2 infection in vivo. Our findings suggest the potential to develop XG014 as pan-β-CoV-B therapeutics and the importance of the XG014 conserved antigenic epitope for designing broadly protective vaccines against β-CoV-B and newly emerging SARS-CoV-2 variants of concern.
Angiotensin-Converting Enzyme 2 ; Antibodies, Neutralizing ; Antibodies, Viral ; COVID-19 ; Epitopes ; Humans ; SARS-CoV-2/genetics* ; Spike Glycoprotein, Coronavirus/genetics*