Objective To explore the mechanism by which puerarin alleviates the cardiotoxicity induced by doxorubicin in myocardial cells. Methods Cells in the logarithmic growth phase were divided into normal control group,model group,low-(20 mmol·L-1),medium-(40 mmol·L-1) and high-(80 mmol·L-1) dose puerarin groups,and positive control group(captopril,1 mmol·L-1). Except for the normal control group,the other groups were co-incubated with 5 mmol·L-1 doxorubicin. Cell viability was assessed using CCK-8 and lactate dehydrogenase (LDH) assays. ROS levels were detected using a ROS probe. Autophagy flux was detected by transfection with HBAD-mcherry-EGFP-LC3 adenovirus. Western Blot was used to measure the protein expression levels of Beclin-1,LC3,p62,p-AMPKα,and AMPKα. Lysosomal function was assessed using a lysosomal probe. Immunofluorescence was used to detect the relative intensity and co-localization of ASMase and LAMP1. Molecular docking analysis was performed to predict the binding capacity of PUE with ASMase. Differential gene expression was analyzed by gene set enrichment analysis. Results Compared to the normal control group,the model group showed reduced cell viability (P<0.01),increased release levels of LDH and ROS (P<0.05,P<0.01),increased number of autophagosomes (P<0.01),and decreased number of autophagic lysosomes (P<0.05). Beclin-1 protein expression and LC3-II/LC3-I ratio decreased(P<0.01),but p62 protein expression increased(P<0.01). Fluorescence intensity of lysosome decreased(P<0.01),whereas fluorescence intensity of ASMase increased(P<0.01). Immunofluorescence co-localization of ASMase and LAMP1 increased (P<0.01),the ratio of p-AMPKα/AMPKα decreased(P<0.05). Compared to the model group,the high-dose puerarin group showed a rebound in cell viability (P<0.05). The medium-and high-dose puerarin groups showed a decreasing trend in LDH level (P<0.05),and all puerarin groups showed a decreasing trend in ROS level (P<0.01). The number of autophagosomes in high-dose puerarin group reduced (P<0.01). The number of autophagic lysosomes in all puerarin groups increased (P<0.05,P<0.01). The high-dose puerarin group showed increased expression of Beclin-1 (P<0.05) and LC3-II/LC3-I ratio,and decreased p62 expression (P<0.01). All puerarin groups showed increased lysosomal fluorescence intensity (P<0.05,P<0.01). The medium-and high-dose puerarin groups showed a decrease in ASMase fluorescence intensity(P<0.05),a reduction in the immunofluorescence co-localization of ASMase with LAMP1 (P<0.01),and an increase in the p-AMPKα/AMPKα ratio (P<0.01). Molecular docking analysis discovered puerarin showed a binding energy of-8.6 kcal·mol-1 with ASMase. Gene enrichment analysis indicated that the differentially expressed genes in the doxorubicin cardiotoxicity model were related to apoptosis,autophagy,and lysosomal function. Conclusion Puerarin can alleviate doxorubicin-induced cardiotoxicity in myocardial cells and protect myocardial cells by regulating autophagy through AMPK/ASMase,as well as restoring autophagic flux.

Objective To investigate the effect of staphylococcal nuclease and tudor domain containing 1(SND1)on the biological function of osteosarcoma cells and decipher the mechanism of SND1 in regulating fer-roptosis in osteosarcoma cells via SLC7A11.Methods Human osteoblasts hFOB1.19 and osteosarcoma cell lines Saos-2,U2OS,HOS,and 143B were cultured,in which the expression level of SND1 was determined.Small in-terfering RNA was employed to knock down the expression of SND1(si-SND1)in the osteosarcoma cell line HOS and 143B.The CCK8 assay kit,colony formation assay,and Transwell assay were employed to examine the effect of SND1 expression on the biological function of osteosarcoma cells.Furthermore,we altered the expression of SND1 and SLC7A11 in osteosarcoma cells to investigate the effect of SND1 on osteosarcoma ferroptosis via SLC7A11.Results The mRNA and protein levels of SND1 in Saos-2,U2OS,HOS,and 143B cells were higher than those in hFOB1.19 cells(all P<0.01).Compared with the control group,transfection with si-SND1 down-regulated the expression level of SND1 in HOS and 143B cells(all P<0.01),decreased the viability of HOS and 143B cells,reduced the number of colony formation,and inhibited cell invasion and migration(all P<0.001).The ferroptosis inducer Erastin promoted the apoptosis of HOS and 143B cells,while the ferroptosis inhibitor Fer-rostatin-1 improved the viability of HOS and 143B cells(all P<0.001).After SND-1 knockdown,Erastin reduced the viability of HOS and 143B cells,while Ferrostatin-1 restored the cell viability(all P<0.001).After treatment with Erastin in the si-SND1 group,the levels of iron and malondialdehyde were elevated,and the level of glutathione was lowered(all P<0.001).The results of in vivo experiments showed that SND1 knockdown inhibited the mass of the transplanted tumor in 143B tumor-bearing nude mice(P<0.001).Knocking down the expression of SND1 resul-ted in down-regulated SLC7A11 expression(all P<0.001)and increased ferroptosis in HOS and 143B cells(P<0.001,P=0.020).Conclusions SND1 presents up-regulated expression in osteosarcoma cells.It may inhibit ferrop-tosis by up-regulating the expression of SLC7A11,thereby improving the viability of osteosarcoma cells.

Humans ; Early Detection of Cancer ; Endoscopy ; Esophageal Neoplasms/epidemiology* ; Risk Factors ; Mass Screening

This study was designed to investigate the effect of silencing microtubule-affinity regulating kinase 4(MARK4)on the apoptosis,inflammatory cytokine release and intestinal barrier protein expression of FHC cells in a lipopolysaccharide(LPS)-induced ulcerative colitis(UC)model,and the underlying molecular mechanisms.Western blot analysis was used to measure the expression levels of MARK4 and apoptosis-related factors including Caspase-1,NLRP3,and GSDMD in colon tissues from both UC patients and healthy individuals,as well as in LPS-induced FHC cell inflammation model.FHC cells was transfected with shRNA to silence MARK4.In control(normal FHC cells),LPS(LPS-stimulated FHC cells),and MARK4-silenced+LPS(shRNA-and LPS-treated FHC cells)groups,the expression levels of Caspase-1,NLRP3,GSDMD,intestinal barrier proteins,and NF-κB pathway-related proteins were assessed by Western blotting.ELISA and RT-qPCR were used to measure the expression levels of inflammatory cytokines IL-1β,IL-6,and TNF-α;flow cytometry was utilized to assess apoptosis.Data showed that both in UC patient colon tissues and the in vitro LPS-induced FHC cell UC inflammation model,there was a significant increase in the expression of MARK4 and apoptosis-related proteins including NLRP3,Caspase-1,and GSDMD.Silencing MARK4 inhibited the expression of these apoptosis-related proteins and downregulated the inflammatory cytokines IL-1β,IL-6,and TNF-α in LPS-induced FHC cells.Silencing MARK4 also reduced apoptosis,increased the expression of intestinal barrier proteins ZO-1,Occludin,and upregulated Claudin2.Gene Set Enrichment Analysis(GSEA)indicated a positive correlation between MARK4 and the NF-κB signaling pathway.Furthermore,silencing of MARK4 inhibited the expression levels of p-P65 and p-IKKα in the NF-κB pathway.In conclusion,MARK4 is significantly upregulated in UC tissues and cells.Silencing MARK4 inhibits the activation of the NF-κB signaling pathway,thereby inhibiting the apoptosis and inflammatory factor expression of UC cells.Thus,MARK4 could be a potential therapeutic target for UC patients.

Objective This clinical study aimed to evaluate the accuracy of a fully digital technique for mea-suring sagittal condylar inclination(SCI),as well as vali-dating whether differences existed between the left and right SCI values of the same participant,to provide a ref-erence for clinical practice.Methods Ten participants with good occlusal relationship and normal temporomandibular joint were recruited.Three methods were used to mea-sure the SCI values of the participants,namely,A(mechanical facebow transferring and mechanical articulator-based measuring method with physical protrusive interocclusal registration),B(face scan-based virtual facebow and virtual ar-ticulator-based measuring method with digital protrusive interocclusal registration),and C(jaw motion tracking system-based measuring method).With the group subjected to methods A and C as the control,the SCI values obtained by the three methods were statistically analyzed.The left and right SCI values of the same participant were also compared.Re-sults The left and right SCI values measured by method A were 41.70°±7.09° and 42.80°±8.62°,those by method B were 35.09°±12.49° and 37.63°±12.10°,and those by method C were 39.43°±8.72° and 38.45°±6.91°.No significant dif-ference existed among the SCI values measured by the three methods(P>0.05).Meanwhile,no statistical difference existed between the SCI values on the left and right sides of the same participant(P>0.05).Conclusion The accuracy of the virtual facebow and digital protrusive occlusal registration based SCI measuring method was the same as that of mechanical facebow based and jaw motion tracking system-based methods.The SCI values on the left and right sides of the same participant were similar.Clinically,an appropriate SCI measurement and setting strategy can be selected based on the actual situations.

Objective This clinical study aimed to as-sess the trueness of three intraoral scanners for the recor-ding of the maximal intercuspal position(MIP)to provide a reference for clinical practice.Methods Ten participants with good occlusal relationship and healthy temporomandibular joint were recruited.For the control group,facebow transferring procedures were performed,and bite registrations at the MIP were used to transfer maxillary and mandibular casts to a mechanical articulator,which were then scanned with a laboratory scanner to obtain digital cast data.For the experimental groups,three intraoral scanners(Trios 3,Carestream 3600,and Aoralscan 3)were used to obtain digital casts of the participants at the MIP following the scanning workflows endorsed by the corresponding manufacturers.Sub-sequently,measurement points were marked on the control group's digital casts at the central incisors,canines,and first molars,and corresponding distances between these points on the maxillary and mandibular casts were measured to calcu-late the sum of measured distances(DA).Distances between measurement points in the incisor(DI),canine(DC),and first molar(DM)regions were also calculated.The control group's maxillary and mandibular digital casts with the added mea-surement points were aligned with the experimental group's casts,and DA,DI,DC,and DM values of the aligned control casts were determined.Statistical analysis was performed on DA,DI,DC,and DM obtained from both the control and ex-perimental groups to evaluate the trueness of the three intraoral scanners for the recording of MIP.Results In the con-trol group,DA,DI,DC,and DM values were(39.58±6.40),(13.64±3.58),(14.91±2.85),and(11.03±1.56)mm.The Trios 3 group had values of(38.99±6.60),(13.42±3.66),(14.55±2.87),and(11.03±1.69)mm.The Carestream 3600 group showed values of(38.57±6.36),(13.56±3.68),(14.45±2.85),and(10.55±1.41)mm,while the Aoralscan 3 group had val-ues of(38.16±5.69),(13.03±3.54),(14.23±2.59),and(10.90±1.54)mm.Analysis of variance revealed no statistically sig-nificant differences between the experimental and control groups for overall deviation DA(P=0.96),as well as local devi-ations DI(P=0.98),DC(P=0.96),and DM(P=0.89).Conclusion With standardized scanning protocols,the three intra-oral scanners demonstrated comparable trueness to traditional methods in recording MIP,fulfilling clinical requirements.

BackgroundPatients with depressive disorder commonly experience sleep disturbances. Previous studies have indicated that group exercise therapy is beneficial in alleviating depressive symptom among patients with depressive disorder. However， there is a lack of research on the impact of group exercise therapy on improving sleep quality in patients with depressive disorder. ObjectiveTo explore the impact of group exercise therapy on sleep quality in patients with acute mild-to-moderate depression during the acute phase， so as to provide references for clinically improving the sleep quality of patients with mild to moderate depressive disorder during the acute phase. MethodsFrom December 2018 to July 2021， patients with mild-to-moderate depressive disorder during the acute phase （n=40）， who met the diagnostic criteria for depressive disorder according to International Classification of Diseases， tenth edition （ICD-10） ，were recruited from the outpatient clinic of Beijing Anding Hospital， Capital Medical University. All participants underwent an 8-week moderate-intensity group exercise therapy program comprising three sessions per week， each lasting 60 minutes. Assessments were conducted at baseline and after 2， 4， 6 and 8 weeks of intervention using Visual Analogue Scale （VAS）， Hamilton Depression Scale-17 item （HAMD-17） and Pittsburgh Sleep Quality Index （PSQI）. The reduction scores at each time point relative to baseline treated as the dependent variables， time as the independent variable， baseline scores as covariates， with time as a fixed effect and baseline values as random effects. Data were analyzed using a linear mixed-effects model. ResultsThe PSQI scores of patients at baseline， 2， 4 ， 6 and 8 weeks after the intervention were （10.62±5.12）， （9.07±3.58）， （7.39±3.66）， （6.54±3.84） and （5.50±3.41）， respectively. The results of linear mixed effect model analysis showed that after 2， 4， 6 and 8 weeks of intervention， patients scored lower than baseline， with statistically significant differences observed in all cases （P<0.01）. The HAMD-17 sleep fcctor scores at baseline， 2， 4， 6 and 8 weeks were （2.25±1.56）， （2.06±1.49）， （1.36±1.27）， （1.22±1.46） and （0.97±1.34）， respectively. The results of linear mixed effects model analysis showed that the HAMD-17 sleep factor scores of 4， 6 and 8 weeks of intervention were lower than that of baseline， and the difference was statistically significant （P<0.05 or 0.01）. The VAS scores at baseline， 2， 4， 6 and 8 weeks after the intervention were （3.18±2.17）， （4.74±2.22）， （6.01±2.31）， （6.54±2.16） and （7.90±1.64）， respectively. The results of linear mixed effect model analysis showed that VAS scores of 2， 4， 6 and 8 weeks of intervention were higher than baseline，and the difference was statistically significant （P<0.01）. ConclusionGroup exercise therapy may improve sleep quality and alleviate depressive symptoms in patients with mild-to-moderate depressive disorder during the acute phase. ［Funded by National Key Research and Development Plan Project （number， 2016YFC1307200）； Beijing Municipal Hospital Scientific Research and Cultivation Plan Project （number， PX2024070）］

Alanine-serine-cysteine transporter 2 (ASCT2) is reported to participate in the progression of tumors and metabolic diseases. It is also considered to play a crucial role in the glutamate-glutamine shuttle of neuroglial network. However, it remains unclear the involvement of ASCT2 in neurological diseases such as Parkinson's disease (PD). In this study, we demonstrated that high expression of ASCT2 in the plasma samples of PD patients and the midbrain of MPTP mouse models is positively correlated with dyskinesia. We further illustrated that ASCT2 expressed in astrocytes rather than neurons significantly upregulated in response to either MPP⁺ or LPS/ATP challenge. Genetic ablation of astrocytic ASCT2 alleviated the neuroinflammation and rescued dopaminergic (DA) neuron damage in PD models in vitro and in vivo. Notably, the binding of ASCT2 to NLRP3 aggravates astrocytic inflammasome-triggered neuroinflammation. Then a panel of 2513 FDA-approved drugs were performed via virtual molecular screening based on the target ASCT2 and we succeed in getting the drug talniflumate. It is validated talniflumate impedes astrocytic inflammation and prevents degeneration of DA neurons in PD models. Collectively, these findings reveal the role of astrocytic ASCT2 in the pathogenesis of PD, broaden the therapeutic strategy and provide a promising candidate drug for PD treatment.

Molecular glues can specifically induce aggregation between two or more proteins to modulate biological functions. In recent years, molecular glues have been widely used as protein degraders. In addition, however, molecular glues play a variety of vital roles, such as complex stabilization, interactome modulation and transporter inhibition, enabling challenging therapeutic targets to be druggable and offering an exciting novel approach for drug discovery. Since most molecular glues are identified serendipitously, exploration of their systematic discovery and rational design are important. In this review, representative examples of molecular glues with various physiological functions are divided into those mediating homo-dimerization, homo-polymerization and hetero-dimerization according to their aggregation modes, and we attempt to elucidate their mechanisms of action. In particular, we aim to highlight some biochemical techniques typically exploited within these representative studies and classify them in terms of three stages of molecular glue development: starting point, optimization and identification.

Lipocalin-2 (LCN2) is a secreted glycoprotein originally purified from mouse kidney cells infected with simian virus 40 and plays a key role in the control of cellular homeostasis during inflammation and the response to cellular stress or injury, and it is considered a potential biomarker for rheumatic diseases, cancer, liver diseases, and inflammatory diseases. Studies have shown that LCN2 is expressed in hepatic parenchymal and nonparenchymal cells and is secreted into the bloodstream, and it is closely associated with the development and progression of acute liver injury, liver cirrhosis, viral hepatitis, alcoholic liver disease, nonalcoholic fatty liver disease, and hepatocellular carcinoma. This article summarizes the animal experiments and clinical studies on the association of LCN2 with the pathogenesis of liver diseases, in order to provide new ideas and therapeutic targets for the prevention and treatment of liver diseases.