Objective:To discuss the enhancing effect of cordycepin on ferroptosis inducer RSL3-induced ferroptosis in the hepatocellular carcinoma HepG2 cells,and to clarify its potential mechanism.Methods:The HepG2 cells were divided into control group,RSL3 group,low,medium and high doses of cordycepin groups,RSL3+low,medium and high doses of cordycepin groups,RSL3+medium-dose cordycepin+ferroptosis inhibitor Ferrostatin-1(Fer-1)group,and RSL3+medium-dose cordycepin+ferroptosis inhibitor Liproxstatin-1(Lip-1)group.The HepG2,Huh-7 and HCCLM3 cells were treated with 0,1,5,10,15 and 20 μmol·L-1 RSL3 for 24,48 and 72 h,respectively.Cell counting kit-8(CCK-8)method was used to detect cell viability and determine the optimal concentration and treatment time of RSL3.The HepG2 cells were treated with 0,50,100,200,400,600,800,1 000,and 1 200 μmol·L-1 cordycepin for 24,48 and 72 h,respectively.CCK-8 method was used to detect the survival rate of the cells and the half maximal inhibitory concentration(IC50)was calculated to determine the optimal concentration and treatment time of cordycepin;the apoptosis inhibitor Z-VAD-FMK,autophagy inhibitor Chloroquine(CQ),necroptosis inhibitor Necrostatin-1(Nec-1),Fer-1,Lip-1,Deferasirox and 2,2,6,6-tetramethylpiper idinoxy(TEMPO)were used to treat HepG2 cells,and the survival rate of the cells was calculated;2',7'-Dichlorodihydrofluorescein diacetate(DCFH-DA)fluorescence probe was used to detect reactive oxygen species(ROS)levels in the HepG2 cells in various groups;C11 BODIPY 581/591 fluorescence probe was used to detect lipid peroxidation(LPO)levels in the HepG2 cells in various groups;FeRhoNox-1 fluorescent probe was used to detect ferrous ion(Fe2+)levels in the HepG2 cells in various groups;kits were used to detect glutathione(GSH)and malondialdehyde(MDA)levels in the HepG2 cells;Western blotting method was used to detect the expression levels of ferroptosis-related proteins,nuclear factor erythroid 2-related factor 2(Nrf2)and heme oxygenase-1(HO-1)proteins in the hepG2 cells in various groups;transmission electron microscope was used to observe the ultrastructural morphology of the HepG2 cells in various groups.Results:The CCK-8 results showed that when the cells were treated with 0.56 μmol·L ?1 RSL3,the viabilities of the three cell types differed significantly.Compared with 0 μmol·L?1 RSL3 group,the survival rates of the cells in 6.4 and 12.8 μmol·L-1 RSL3 groups were significantly decreased(P<0.05).The HepG2 cells had the highest IC50 value and were selected for subsequent experiments.Compared with 0 μmol·L-1 cordycepin group,the survival rates of the HepG2 cells in 200,400,600,800,1 000,and 2 000 μmol·L-1 cordycepin groups were significantly decreased(P<0.05 or P<0.01).0.5×IC50(267.9 μmol·L-1),1×IC50(535.8 μmol·L-1)and 1.5×IC50(803.7 μmol·L-1)were selected as low,medium and high doses of cordycepin groups,respectively,with an intervention time of 24 h.Compared with control group,the survival rates of the HepG2 cells in low,medium,and high doses of cordycepin groups were significantly decreased(P<0.05).Compared with low,medium,and high doses of cordycepin groups,the survival rates of the HepG2 cells in Z-VAD-FMK+low,medium,and high doses of cordycepin groups were significantly increased(P<0.01),and those in Fer-1+medium and high doses of cordycepin groups and Lip-1+low,medium,and high doses of cordycepin groups were significantly increased(P<0.05).Compared with control group,the survival rates of the HepG2 cells in RSL3 group,RSL3+low,medium,and high doses of cordycepin groups,RSL3+cordycepin+Fer-1 group and RSL3+cordycepin+Lip-1 group were significantly decreased(P<0.05 or P<0.01).Compared with RSL3 group,the survival rates of the HepG2 cells in RSL3+low,medium,and high doses of cordycepin groups were significantly decreased(P<0.05).The DCFH-DA results showed that compared with control group,the ROS levels in the cells in medium and high doses of cordycepin groups,RSL3 group and RSL3+low,medium,and high doses of cordycepin groups were significantly increased(P<0.05 or P<0.01).The C11 BODIPY 581/591 results showed that compared with control group,the LPO levels in the HepG2 cells in medium and high doses of cordycepin groups,RSL3 group and RSL3+low,medium and high doses of cordycepin groups were significantly increased(P<0.05 or P<0.01).Compared with RSL3 group,the LPO levels in the HepG2 cells in RSL3+low,medium,and high doses of cordycepin groups were significantly increased(P<0.05 or P<0.01).The FeRhoNox-1 results showed that compared with control group,the Fe2+levels in the HepG2 cells in medium and high doses of cordycepin groups,RSL3 group and RSL3+low,medium,and high doses of cordycepin groups were significantly increased(P<0.05 or P<0.01).Compared with RSL3 group,the Fe2+levels in the HepG2 cells in RSL3+low,medium,and high doses of cordycepin groups were significantly increased(P<0.05 or P<0.01).Compared with control group,the MDA levels in the HepG2 cells in high doses of cordycepin group,RSL3 group and RSL3+low,medium,and high doses of cordycepin groups were significantly increased(P<0.05 or P<0.01).Compared with RSL3 group,the MDA levels in the HepG2 cells in RSL3+low,medium,and high doses of cordycepin groups were significantly increased(P<0.05 or P<0.01).Compared with control group,the GSH levels in the HepG2 cells in medium and high doses of cordycepin groups,RSL3 group and RSL3+low,medium,and high doses of cordycepin groups were significantly decreased(P<0.05 or P<0.01).Compared with RSL3 group,the GSH levels in the HepG2 cells in RSL3+low,medium,and high doses cordycepin groups were significantly decreased(P<0.05 or P<0.01).Compared with control group,the ultrastructure of the HepG2 cells in low and medium doses of cordycepin groups showed no significant changes,while the cells in high dose of cordycepin group exhibited reduced mitochondrial cristae,mild swelling and increased membrane density,with slightly distorted inner membrane structure.The cells in RSL3 group and RSL3+low,medium,and high doses of cordycepin groups all showed ultrastructural changes characteristic of ferroptosis.Compared with RSL3 group,the cells in RSL3+low,medium,and high doses of cordycepin groups exhibited ruptured mitochondrial membranes with increased membrane density,abnormally twisted or expanded inner membrane structures,and reduced or even disappeared mitochondrial cristae.The Western blotting results showed that compared with control group,the expression levels of FTH1 and GPX4 proteins in the HepG2 cells in medium and high doses of cordycepin groups were significantly decreased(P<0.05 or P<0.01),while the expression levels of Nrf2 and HO-1 proteins were significantly decreased(P<0.05 or P<0.01).Compared with control group,the expression levels of GPX4 protein in the HepG2 cells in low,medium and high doses of cordycepin groups,RSL3 group,and RSL3+cordycepin+Fer-1 group were significantly decreased(P<0.05).Compared with RSL3 group,the expression levels of GPX4 protein in the HepG2 cells in RSL3+low,medium,and high doses of cordycepin groups were significantly decreased(P<0.05).Conclusion:Cordycepin can significantly enhance RSL3-induced ferroptosis in the hepatocellular carcinoma HepG2 cells and down-regulate the expression of Nrf2 and HO-1 proteins in the HepG2 cells.

Objective To investigate the regulatory effect of circular RNA ASAP1(circASAP1)targeting microRNA-4500(miR-4500)on the proliferation and apoptosis of osteosarcoma cells.Methods Human osteosarcoma cells(Saos-2)were selected as the experimental subjects and cul-tured in DMEM medium containing 10％fetal bovine serum while maintaining 5％CO2.The expres-sions of circASAP1 and miR-4500 in osteosarcoma tissues were detected using RT-qPCR.Saos-2 cells were transfected with si-NC(40 nmol/L),si-circASAP1(40 nmol/L),pcDNA(0.4 μg),pcDNA-circASAP1(0.4 μg),miR-NC(40 nmol/L),miR-4500 mimics(40 nmol/L),si-circASAP1+anti-miR-NC(40 nmol/L),and si-circASAP1+anti-miR-4500(40 nmol/L),respectively.Cell proliferation and apoptosis were assessed using CCK-8,colony formation assays,and flow cytometry.The interaction between circASAP1 and miR-4500 was analyzed using a dual-luciferase reporter assay.Results Compared with adjacent non-tumor tissues,the expression of circASAP1 was significantly upregulated,while that of miR-4500 was significantly downregulated in osteosarcoma tissues(P＜0.001).Compared with the si-NC group,the si-circASAP1 group exhibited significantly decreased circASAP1 expression,optical density(OD)values,and the number of cell colonies(P＜0.001).Compared with the si-NC group,the si-circASAP1 group showed significantly upregulated levels of cleaved-caspase3 protein,cleaved-caspase9 protein,and the apoptosis rate(P＜0.001).StarBase search revealed specific binding sequences between miR-4500 and circASAP1.Co-transfection of miR-4500 mimics and WT-circASAP1 significantly reduced the relative luciferase activity of the cells[(0.34±0.03)versus(0.95±0.06),t=27.280,P＜0.001].The expression of miR-4500 in the pcDNA-circASAP1 group was significantly lower than that in the pcDNA group,whereas the ex-pression of miR-4500 in the si-circASAP1 group was significantly higher than that in the si-NC group(P＜0.001).Compared with the miR-NC group,the miR-4500 group exhibited significantly in-creased miR-4500 expression,cleaved-caspase3 protein levels,cleaved-caspase9 protein levels,and the apoptosis rate,while the OD values and the number of colonies significantly decreased(P＜0.001).Compared with the si-circASAP1+anti-miR-NC group,the si-circASAP1+anti-miR-4500 group showed significantly decreased miR-4500 expression,cleaved-caspase3 protein levels,cleaved-caspase9 protein levels,and the apoptosis rate,while the OD values and the number of col-onies significantly increased(P＜0.001).Conclusion The expression of circASAP1 is increased,while that of miR-4500 is decreased in osteosarcoma tissues.Moreover,circASAP1 promotes the proliferation and inhibits the apoptosis of osteosarcoma cells by targeting miR-4500.

ObjectiveTo investigate the protective effect of cytochrome P4502D6 （CYP2D6） and cytochrome P4503A4 （CYP3A4）， key enzymes of drug metabolism in liver， on acute liver injury in water extract of Glycyrrhizae Radix et Rhizoma （WEOGRR）. MethodHealthy male Kunming mice were divided into normal group， model group， WEOGRR low-， medium- and high-dose groups （5， 10， 15 g·kg^-1·d^-1） and positive drug group （diammonium glycyrrhizinate， 75 mg·kg^-1·d^-1）， with 10 in each group. One week after preventive administration， acute liver injury model was induced by single intragastric administration of 270 mg·kg^-1 Tripterygium Glycosides tablets， and samples were collected after 18 h. The pathological changes of liver were observed by hematoxylin-eosin （HE） staining. Serum liver function indexes including alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， γ-glutamyl transpeptadase （γ-GT）， alkaline phosphatase （ALP）， and total bilirubin （TBIL） as well as the levels of oxidative stress indexes including malondialdehyde （MDA） and superoxide dismutase （SOD） in hepatocytes were determined by biochemical method. Real-time polymerase chain reaction （Real-time PCR） and Western blot were performed to detect the mRNA and protein expression levels of CYP2D6 and CYP3A4， respectively. ResultCompared with normal group， model group had significant hepatocyte swelling and inflammatory cell infiltration （P<0.01）， increased AST， ALT， γ-GT， ALP and TBIL （P<0.05）， elevated MDA and decreased SOD （P<0.01） as well as down-regulated mRNA and protein expression levels of CYP2D6 and CYP3A4 （P<0.05）. Compared with the model group， the normal group had intact liver structure without obvious abnormality， and the WEOGRR groups and positive drug group presented alleviated hepatocyte swelling and inflammatory cell infiltration （P<0.01）， reduced AST， ALT， γ-GT， ALP and TBIL （P<0.01）， lowered MDA and increased SOD （P<0.01） as well as up-regulated expression levels of CYP2D6 and CYP3A4 （P<0.01）. ConclusionThe protective effect of WEOGRR on acute liver injury induced by Tripterygium glycosides tablets may be related to reducing the contents of AST， ALT， γ-GT， ALP and TBIL in serum， inhibiting MDA and increasing the activity of SOD in liver cells， and enhancing the activities of CYP2D6 and CYP3A4， thus accelerating the metabolism of toxic substances.

Objective:To compare the effects of low-carbohydrate diet (LCD) and low-fat diet (LFD) in the lifestyle intervention of non-alcoholic fatty liver disease (NAFLD) through a meta-analysis of randomized controlled trials.Methods:PubMed, Embase, Web of Science, Cochrane, CNKI and Wanfang were searched for relevant studies and study references and conference proceedings were manually searched. Two authors independently screened the items retrieved, extracted the data and assessed the quality of included studies. Meta-analysis was performed using R4.4.1 and RevMan5.4.1. Data were pooled using random-effects models and potential sources of heterogeneity were investigated using stratified meta-analysis. Funnel plots and Peters' test were used to assess publication bias.Results:Nine studies with a total of 510 participants met our inclusion criteria. Meta-analysis results showed that LCD and LFD interventions had similar effects on the reduction of intrahepatic lipid content in NAFLD patients ( SMD: -0.31,95% CI: 0.97 to 0.35, P = 0.36). There were no significant differences in changes of alanine aminotransferase ( SMD: -0.25, 95%CI: 0.91 to 0.41, P = 0.45) and aspartate aminotransferase ( SMD: -0.45, 95%CI: 1.63 to 0.72, P = 0.45) levels, either. Subgroup analyses implied that the duration of different interventions might be the cause of heterogeneity across studies. No significant publication bias was showed in the meta-analysis. Conclusion:Current evidence from randomized controlled studies does not support the superiority of LCD over LFD in the treatment of NAFLD.

Objective:To analyze the research hotspots and frontiers of hospital research management research from 1981 to 2022.Methods:The relevant literature in the field of hospital scientific research management was retrieved from the CNKI database to explore the trends of publications in this research field. A scientific knowledge graph was drawn and a visualization analysis of the information of authors, issuing units, and research institutions were conducted by Cite Space.5.8.R3.Research hotspots were discussed based on keyword emergence, cluster analysis, and keyword time zone graph.Results:The publication trend in this field was generally policy-oriented, but the cooperation among authors and institutions was relatively loose, and the research hotspots were gradually shifting from scientific research funding management to discipline construction, talent training, translational medicine, and informatization. Cluster analysis found that the main content of hospital scientific research management was scientific research funding and clinical scientific research management and the main management objects were the medical and nursing staff.Conclusions:Hospital scientific research managers must adhere to the policy-oriented approach, strengthen the cooperation and exchanges in scientific research management, innovate the scientific research management mode around the research hotspots and development trends, and promote the quality and efficiency of scientific research management.

【Objective】 To study the genotypes of ABO ambiguous blood group samples(n=20) and identify their molecular biological characteristics. 【Methods】 The serological phenotype of the samples was analyzed by serological techniques. Seven exons of ABO gene were amplified by polymerase chain reaction (PCR) and the PCR products were directly sequenced; the genotypes and sequences of ABO subtypes were analyzed. 【Results】 The serological phenotypes of 20 samples presenting ABO ambiguous blood group were as follows: weak A antigen (n=5), weak A antigen combined with anti-A1 antibody (n=5), normal A antigen combined with anti-A1 antibody (n=2), weak B antigen (n=8). The genotypes of them were as follows: Ax02/O01 (n=3), Ael07/O01 (n=2), B313/O01 (n=2), A204/O02 (n=1), A220/O01 (n=1), Ael07/O02 (n=1), Ael02/O01 (n=1), Ael02/O02 (n=1), Ax03/O01 (n=1), Ax03/O02 (n=1), B313/O02 (n=1), B302/O01 (n=1), B302/O02 (n=1), Bw19/O02 (n=1), A102/B313 (n=1) and A101/Bw37 (n=1). 【Conclusion】 ABO genotyping technology can accurately identify the ambiguous blood group of samples, provide definite genetic information of blood group and ensure the safety of clinical transfusion.

Tripterygium glycosides tablet(TGT),the classical commercial drug of Tripterygium wilfordii Hook.F.has been effectively used in the treatment of rheumatoid arthritis,nephrotic syndrome,leprosy,Behcet's syndrome,leprosy reaction and autoimmune hepatitis.However,due to its narrow and limited treatment window,TGT-induced organ toxicity(among which liver injury accounts for about 40％of clinical reports)has gained increasing attention.The present study aimed to clarify the cellular and molecular events underlying TGT-induced acute liver injury using single-cell RNA sequencing(scRNA-seq)technology.The TGT-induced acute liver injury mouse model was constructed through short-term TGT exposure and further verified by hematoxylin-eosin staining and liver function-related serum indicators,including alanine aminotransferase,aspartate aminotransferase,alkaline phosphatase and total bilirubin.Using the mouse model,we identified 15 specific subtypes of cells in the liver tissue,including endothelial cells,hepatocytes,cholangiocytes,and hepatic stellate cells.Further analysis indicated that TGT caused a significant inflammatory response in liver endothelial cells at different spatial locations;led to marked inflammatory response,apoptosis and fatty acid metabolism dysfunction in hepatocytes;activated he-patic stellate cells;brought about the activation,inflammation,and phagocytosis of liver capsular macrophages cells;resulted in immune dysfunction of liver lymphocytes;disturbed the intercellular crosstalk in liver microenvironment by regulating various signaling pathways.Thus,these findings elaborate the mechanism underlying TGT-induced acute liver injury,provide new insights into the safe and rational applications in the clinic,and complement the identification of new biomarkers and ther-apeutic targets for liver protection.

Objective:To study the effect of sacubitril valsartan sodium tablets on serum tenascin-C (TN-C) level and myocardial remodeling in patients of chronic left heart failure (CHF) complicated with renal failure.Methods:A total of 84 patients with chronic left heart failure complicated with renal failure admitted to Qinhuangdao Jungong Hospital from October 2020 to October 2021 were included and divided into the observation group (treated with sacubitril valsartan sodium tablets) and the control group (treated with valsartan), with 42 cases in each group according to the random number table method. The clinical efficacy of the two groups was compared after 3 months of treatment. The TN-C level and cardiac function index left ventricular end-diastolic diameter (LVEDD), left ventricular ejection fraction (LVEF), troponin T (cTnT) and other index before the treatment and after 3 months of treatment were compared between the two groups.Results:After 3 months of treatment, the total effective rate between the two groups had no significant difference ( P>0.05). After 3 months of treatment, the TN-C level in the observation group was lower than that in the control group: (32.42 ± 4.22) μg/L vs. (37.32 ± 4.86) μg/L; and the LVEF in the observation group was higher than that in the control group: (41.21 ± 5.39)% vs. (37.76 ± 5.45)%, the differences were statistically significant ( P<0.05). The LVEDD and cTnT in the two groups had no significant differences ( P>0.05). After 3 months of treatment, neuroendocrine factors norepinephrine, aldosterone, angiotensin Ⅱlevels in the in the observation group were lower than those in the control group: (1 668.60 ± 251.19) pmol/L vs. (2 005.86 ± 280.91) pmol/L, (246.97 ± 13.99) ng/L vs. (275.41 ± 19.38) ng/L, (99.68 ± 8.57) ng/L vs. (112.20 ± 9.52) ng/L, the differences were statistically significant ( P<0.05). Conclusions:Sacubitril valsartan sodium tablets have a good effect in the treatment of CHF complicated with renal failure, which can improve the cardiac function and inhibit the over-activation of neuroendocrine hormones.

Objective: To establish colorectal cancer LoVo and SW620 cell lines that stably interfere with the expression of LINC01224, and to explore the effect of down-regulating the expression of LINC01224 on cell apoptosis. Methods: The GEPIA2 database was used to analyze the expression of LINC01224 in colorectal cancer tissues; qPCR method was used to detect the expression of LINC01224 in 10 human colorectal cancer cells. Three different LINC01224 siRNAs were respectively transfected into human colorectal cancer LoVo cells, and the LINC01224 shRNA lentiviral vector was constructed with the siRNA sequence with the most obvious inhibitory effect of LINC01224 expression. Recombinant lentiviral particles were packaged in HEK293 T cells and then infected with LoVo and SW620 cells. After selection by puromycin, the monoclonal cells that stably interfere with LINC01224 were obtained by limiting dilution method. MTS method detects cell proliferation ability, and flow cytometry detects cell apoptosis rate. Results: The expression of LINC01224 in colorectal cancer tissues was higher than that in normal colorectal tissues, and its expression in 10 types of colorectal cancer cells was also higher than that in normal colorectal epithelial cells HCOEPic. The inhibition rate of siRNA-3 on the expression of LINC01224 in LoVo cells was higher than that of siRNA-1 and siRNA-2. Therefore, siRNA-3 was chosen to design LINC01224 shRNA.Compared with the control group(sh-NC group), the expression level of LINC01224 in the LoVo and SW620 cells of the stable interference LINC01224 group(sh-LINC01224 group) was reduced(P<0.01), and the cell growth rate was slowed down(P<0.01), the rate of apoptosis also increased(P<0.01). Conclusion The shRNA lentiviral interference vector of LINC01224 is successfully constructed, which can stably infect LoVo and SW620 cells, down-regulate the expression of LINC01224 and induce cell apoptosis.

Objective:The aim of this study was to investigate the prognostic value of vitamin B12 as the non-invasive biomarker to predict long-term rebleeding rate in cirrhotic patients with esophagogastric varices.Methods:From Dec 1, 2016 to Dec 31, 2017, cirrhotic patients with esophagogastric varices who had been admitted to Zhongshan Hospital affiliated to Fudan University were enrolled. All these patients received endoscopic treatment to prevent variceal rebleeding. The serum vitamin B12 and folic acid levels were measured in all of them. The receiver operating characteristic (ROC) analysis, Kaplan-Meier analysis, univariate and multivariate cox regression analysis were conducted to explore the value of vitamin B12 in predicting 3-year variceal rebleeding in cirrhotic patients with esophagogastric varices after endoscopic treatment.Results:115 patients were included. The ROC curve analysis indicated that the optimal cutoff value of vitamin B12 for 3-year variceal rebleeding was 567.25 pg/ml. According to the cut-off value, the patients were divided into high-level vitamin B12 group ( n=49) and low-level vitamin B12 group ( n=66). Compared with the low vitamin B12 group, the high vitamin B12 group had lower albumin level, less male (63.3% vs 80.3%), and higher 3-year rebleeding rate ( P<0.05). Cox analysis showed that vitamin B12 and platelet were independent prognostic factors for 3-year rebleeding in patients with variceal bleeding. Conclusions:Elevated peripheral blood vitamin B12 predicts a higher risk of long-term rebleeding in patients with liver cirrhosis and esophagogastric varices.