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.

Objective:To investigate the mode and timing of delivery in pregnant women with complex fetal congenital heart disease (CHD) detected by prenatal ultrasound.Methods:The clinical data of 123 fetuses with complex CHD detected by prenatal ultrasound in Peking University People′s Hospital from January 2016 to December 2023 were retrospectively analyzed. Pregnant women with indications for prenatal diagnosis underwent G-banding karyotype analysis, single nucleotide polymorphism array (SNP arrry) or whole exome sequencing after informed consent. Integrated managements were provided for pregnant women with complex CHD during pregnancy and perinatal period, and to determine the mode and timing of delivery. Infants with complex CHD received timely treatment or referral after birth.Results:The gestational age at ultrasound diagnosis of the 123 fetuses with complex CHD was (23.7±3.4) weeks. There were 11 cases (8.9%) of total anomalous pulmonary venous connection (TAPVC), 14 cases (11.4%) of anomalous pulmonary valve (PVA), 7 cases (5.7%) of right ventricle double outlet (RVDO), 13 cases (10.6%) of anomalous aortic arch, 69 cases (56.1%) of transposition of the great arteries (TGA), 9 cases (7.3%) of other types. All cases were treated with fetal preservation after prenatal consultation. Among the 72 cases undergoing prenatal diagnosis, 9 cases (12.5%) had chromosomal variations. Fifty-five cases (44.7%, 55/123) underwent trial of labor, of which 46 cases (37.4%, 46/123) had successful vaginal delivery, including 6 cases of forceps delivery, and other 9 cases of failed trial of labor transferred to cesarean section. A total of 77 cases of pregnant women underwent cesarean section. Except for the 21 cases of pregnant women who asked for cesarean section, the cesarean section rate of pregnant women with complex CHD was 45.5% (56/123), which was not significantly different from the average cesarean section rate of the same period in our hospital (40.2%; χ2=7.34, P=0.270). The gestational age at delivery of the 123 fetuses with complex CHD was (37.9±1.4) weeks, the birth weight of the neonates was (3 099±480) g, and the umbilical artery blood pH value was 7.31±0.05. The oxygen saturation of 86 cases before and after alprostadil infusion were (72.8±6.0)% and (80.5±5.0)%, respectively, and the difference was statistically significant ( t=4.38, P<0.001). One hundred and fourteen children underwent surgical treatment, 112 of them (98.2%) had good postoperative reexamination. Only 2 cases (1.8%) died after surgery, and 14.0% (16/114) had the possibility of secondary surgery. Conclusions:Fetal complex CHD is not an indication for cesarean section, and the delivery mode could be selected according to the obstetric situation. If the mother and child are in stable condition, the delivery is planned after 39 weeks of gestation. For children with low oxygen saturation after birth, alprostadil could be pumped to maintain the open ductus arteriosus, and timely referral to the pediatric cardiac surgery for subsequent surgical treatment, which could achieve a good prognosis.

Kleine-Levin syndrome (KLS) is a rare, recurrent-remitting sleep disorder characterized by periodic hypersomnia, hyperphagia, and hypersexuality, accompanied by cognitive dysfunction, perceptual disturbances, mood changes, or psychiatric abnormalities. Patients typically experience recurrent episodes, with hypersomnic periods lasting from several days to weeks. Most KLS patients exhibit normal functions during the intermission period, while early-onset KLS cases in adolescents are rare, often presenting with frequent episodes and complex symptoms, which can easily lead to misdiagnosis. Therefore, this case report analyzed the detailed medical history, laboratory findings, differential diagnosis, and treatment of a teenage patient with KLS, along with a review of the literature. This case aims to provide a deeper understanding of the complex symptoms and diagnostic challenges associated with early-onset KLS in adolescents.

Objective:To explore the current status and trends in emergency nursing teaching methods based on CiteSpace software in China from 2012 to 2023, and to provide reference for subsequent research.Methods:Literature on teaching methods in emergency nursing was electronically searched in the China National Knowledge Infrastructure, WanFang Data, and VIP from January 2012 to December 2023. CiteSpace and Excel software were used to perform bibliometric analysis on the publication year, journal source, first author affiliation, keywords and clustering content.Results:An initial search yielded 1 550 articles and 936 relevant articles were screened and published in 194 journals. From 2012 to 2023, the number of publications on teaching methods in emergency nursing in China was relatively flat, the first author affiliations were mainly general hospitals, and the research methods were mainly quantitative. Keyword clustering showed that six themes were extracted, including exploration of professional skills, reform and application of teaching methods, nursing teaching and evaluation, teaching modes, teaching tools, and training modes.Conclusions:The trend of publications in emergency nursing teaching methods in China in the past 10 years has been relatively flat, and the quality of the literature needs to be improved. Emergency nursing teaching needs to focus on reforming teaching modes and methods, and further exploring the use of tools such as mind maps and teaching manuals to improve the quality of teaching.

Objective:To investigate the mode and timing of delivery in pregnant women with complex fetal congenital heart disease (CHD) detected by prenatal ultrasound.Methods:The clinical data of 123 fetuses with complex CHD detected by prenatal ultrasound in Peking University People′s Hospital from January 2016 to December 2023 were retrospectively analyzed. Pregnant women with indications for prenatal diagnosis underwent G-banding karyotype analysis, single nucleotide polymorphism array (SNP arrry) or whole exome sequencing after informed consent. Integrated managements were provided for pregnant women with complex CHD during pregnancy and perinatal period, and to determine the mode and timing of delivery. Infants with complex CHD received timely treatment or referral after birth.Results:The gestational age at ultrasound diagnosis of the 123 fetuses with complex CHD was (23.7±3.4) weeks. There were 11 cases (8.9%) of total anomalous pulmonary venous connection (TAPVC), 14 cases (11.4%) of anomalous pulmonary valve (PVA), 7 cases (5.7%) of right ventricle double outlet (RVDO), 13 cases (10.6%) of anomalous aortic arch, 69 cases (56.1%) of transposition of the great arteries (TGA), 9 cases (7.3%) of other types. All cases were treated with fetal preservation after prenatal consultation. Among the 72 cases undergoing prenatal diagnosis, 9 cases (12.5%) had chromosomal variations. Fifty-five cases (44.7%, 55/123) underwent trial of labor, of which 46 cases (37.4%, 46/123) had successful vaginal delivery, including 6 cases of forceps delivery, and other 9 cases of failed trial of labor transferred to cesarean section. A total of 77 cases of pregnant women underwent cesarean section. Except for the 21 cases of pregnant women who asked for cesarean section, the cesarean section rate of pregnant women with complex CHD was 45.5% (56/123), which was not significantly different from the average cesarean section rate of the same period in our hospital (40.2%; χ2=7.34, P=0.270). The gestational age at delivery of the 123 fetuses with complex CHD was (37.9±1.4) weeks, the birth weight of the neonates was (3 099±480) g, and the umbilical artery blood pH value was 7.31±0.05. The oxygen saturation of 86 cases before and after alprostadil infusion were (72.8±6.0)% and (80.5±5.0)%, respectively, and the difference was statistically significant ( t=4.38, P<0.001). One hundred and fourteen children underwent surgical treatment, 112 of them (98.2%) had good postoperative reexamination. Only 2 cases (1.8%) died after surgery, and 14.0% (16/114) had the possibility of secondary surgery. Conclusions:Fetal complex CHD is not an indication for cesarean section, and the delivery mode could be selected according to the obstetric situation. If the mother and child are in stable condition, the delivery is planned after 39 weeks of gestation. For children with low oxygen saturation after birth, alprostadil could be pumped to maintain the open ductus arteriosus, and timely referral to the pediatric cardiac surgery for subsequent surgical treatment, which could achieve a good prognosis.

Objective:To explore the current status and trends in emergency nursing teaching methods based on CiteSpace software in China from 2012 to 2023, and to provide reference for subsequent research.Methods:Literature on teaching methods in emergency nursing was electronically searched in the China National Knowledge Infrastructure, WanFang Data, and VIP from January 2012 to December 2023. CiteSpace and Excel software were used to perform bibliometric analysis on the publication year, journal source, first author affiliation, keywords and clustering content.Results:An initial search yielded 1 550 articles and 936 relevant articles were screened and published in 194 journals. From 2012 to 2023, the number of publications on teaching methods in emergency nursing in China was relatively flat, the first author affiliations were mainly general hospitals, and the research methods were mainly quantitative. Keyword clustering showed that six themes were extracted, including exploration of professional skills, reform and application of teaching methods, nursing teaching and evaluation, teaching modes, teaching tools, and training modes.Conclusions:The trend of publications in emergency nursing teaching methods in China in the past 10 years has been relatively flat, and the quality of the literature needs to be improved. Emergency nursing teaching needs to focus on reforming teaching modes and methods, and further exploring the use of tools such as mind maps and teaching manuals to improve the quality of teaching.

Kleine-Levin syndrome (KLS) is a rare, recurrent-remitting sleep disorder characterized by periodic hypersomnia, hyperphagia, and hypersexuality, accompanied by cognitive dysfunction, perceptual disturbances, mood changes, or psychiatric abnormalities. Patients typically experience recurrent episodes, with hypersomnic periods lasting from several days to weeks. Most KLS patients exhibit normal functions during the intermission period, while early-onset KLS cases in adolescents are rare, often presenting with frequent episodes and complex symptoms, which can easily lead to misdiagnosis. Therefore, this case report analyzed the detailed medical history, laboratory findings, differential diagnosis, and treatment of a teenage patient with KLS, along with a review of the literature. This case aims to provide a deeper understanding of the complex symptoms and diagnostic challenges associated with early-onset KLS in adolescents.

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.

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.