To explore the inhibitory effect of ginkgolide B (GB) on MH7A human fibroblast-like synoviocytes (FLS) and its potential mechanism. Firstly, 20 μg/L tumor necrosis factor-α (TNF-α) was pretreated with MH7A to establish a cell model of arthritis. After incubation of MH7A cells with various concentrations of GB, CCK-8 assay, Transwell assay, and flow cytometry (FCM) were separately used to detect cell viability, cell invasion, and cell apoptosis rate and cell cycle; Real-time quantitative PCR and Western blot assay were performed to detect the apoptosis- and cycle-related gene transcriptions and protein expressions, respectively. The results showed that compared with the control group, GB dose- and time-dependently suppressed cell viability to a greater extent; GB significantly reduced cell invasive ability and increased cell apoptosis rate and proportion of G0/G1 phase in MH7A cells, along with increased transcription levels of Bcl-2-associated X protein (Bax) and p21 mRNA and decreased transcription levels of Bcl-2, myeloid cell leukemia 1(Mcl-1), protein kinase B (PKB; AKT), IP3K, Cyclin D1 and cyclin-dependent kinase 4 (CDK4) mRNA; GB remarkably increased expression levels of Bax, p21, and cleaved-Caspase 3 protein and decreased expression levels of Bcl-2, Mcl-1, p-AKT, p-PI3K, Cyclin D1, and CDK4 protein, with decreased ratios of p-PI3K/PI3K, p-AKT/AKT, and Bcl-2/Bax. In conclusion, GB blocks the G1-to-S cell cycle transition, suppresses cell viability and cell invasion and induces cell apoptosis of MH7A human RA-FLS via suppressing the PI3K/AKT signaling pathway.

Objective To compare the effects of three kinds of artificial hydronephrosis in percutaneous nephrolithotomy.Methods A total of 120 patients who underwent single-tract percutaneous nephrolithotomy in the Eighth Hospital of Wuhan from May 2020 to April 2022 were selected and divided equally into three groups according to different methods of artificial hydronephrosis.Patients in group A were received preoperative indwelling ureteral catheter and injection of normal saline through ureteral catheter to dilate renal pelvis and form artificial hydronephrosis.Patients in Group B were placed with double J catheters before surgery,and the bladder was filled with physiological saline through the catheter.The renal pelvis was dilated through the double J catheters,resulting in artificial hydronephrosis.Patients in Group C were received intravenous injection of furosemide and stimulated diuretic method to actively dilate renal pelvis to form hydronephrosis.The one-time puncture success rate,channel establishment time,overall operation time,stone clearance rate and incidence of surgical complications of percutaneous nephrolithotomy after hydronephrosis were compared among the three groups.Results The operation was successfully completed in the three groups.There was no significant difference in the one-time puncture success rate and channel establishment time between group A and group B(P>0.05),which were all higher than group C(P<0.05).The overall operation time of group B was shorter than that of group A and group C(P<0.05).There were no significant differences in stone clearance rate and surgical complications among the three groups(P>0.05).Conclusion Preoperative indwelling of double J tubes to create artificial kidney hydronephrosis has advantages such as high success rate of one-time puncture,short channel establishment time,and surgical time.

Objective To investigate the influencing factors of delirium after cardiac valve replacement went under car-diopulmonary bypass(CPB)with propofol sedation.Methods A total of 152 patients underwent cardiac valve replacement under CPB in Nanyang Central Hospital from January 2020 to December 2022 were selected as research objects,and they were randomly divided into observation group A[50 ≤bispectral index(BIS)＜60]and observation group B(35≤BIS＜45)according to the depth of propofol sedation,with 76 cases in each group.The clinical data such as age,gender,body mass index(BMI),diabetes,hypertension,coronary heart disease,chronic obstructive pulmonary disease,sleep disorder,nutritional disorder,anxiety,depression,smoking history,drinking history,preoperative cardiac insufficiency,intraoperative hypoxemia,intraoperative hypoproteinemia,postoperative acute renal injury,secondary intubation,massive blood transfusion,excessive pain,postoperative left ventricular ejection fraction(LVEF),surgical method and CPB time were collected,and the incidence of postoperative delirium of patients was evaluated by the confusion assessment method of intensive care unit(CAM-ICU)method.The incidence of postoperative delirium of patients between observation group A and observation group B was compared.The influencing factors of postoperative delirium occurrence was analyzed by using univariate and multivariate logis-tic regression analysis.Results Among the 152 patients underwent heart valve replacement,36 patients experienced postoperative delirium,with an incidence of 23.68％.The incidence of postoperative delirium of patients in the observation group A and the observation group B was 38.16％(29/76),9.21％(7/76),respectively;the incidence of postoperative delirium of patients in the observation group A was significantly higher than that in the observation group B(x2=17.617,P＜0.05).The gender,BMI,diabetes,hypertension,coronary heart disease,cognitive disorder,sleep disorder,nutritional disorder,anxiety,depression,smoking history,drinking history,intraoperative hypoxemia,intraoperative hypoproteinemia,postoperative acute renal injury,secondary intubation,massive blood transfusion,and surgical method were not related to postoperative delirium(P＞0.05);the age,chronic obstructive pulmonary disease,preoperative heart failure,excessive pain,postoperative LVEF,and CPB time were associated with postoperative delirium(P＜0.05).Multivariate logistic regression analysis showed that age 60 years,preoperative cardiac dysfunction,excessive pain,and CPB time≥100 minutes were risk factors for postoperative delirium(P＜0.05),while postoperative LVEF≤50％and propofol sedation depth of 35≤BIS＜45 were protective factors for postoperative delirium(P＜0.05).Conclusion Propofol sedation depth of 35≤BIS＜45,postoperative LVEF ≥50％can effectively reduce the risk of postoperative delirium after cardiac valve replacement under CPB.Age≥60 years old,preoperative cardiac insufficiency,excessive pain,and CPB time≥100 min can increase the risk of postoperative delirium.

Objective To identify the key genes differentially expressed in Wilms tumor and analyze their potential impacts on prognosis and immune responses of the patients. Methods High-throughput RNA sequencing was used to identify the differentially expressed mRNAs in clinical samples of Wilms tumor and paired normal tissues, and their biological functions were analyzed using GO, KEGG and GSEA enrichment analyses. The hub genes were identified using STRING database, based on which a prognostic model was constructed using LASSO regression. The mutations of the key hub genes were analyzed and their impacts on immunotherapy efficacy was predicted using the cBioPortal platform. RT-qPCR was used to verify the differential expressions of the key hub genes in Wilms tumor. Results Of the 1612 differentially expressed genes identified in Wilms tumor, 1030 were up-regulated and 582 were down-regulated, involving mainly cell cycle processes and immune responses. Ten hub genes were identified, among which 4 genes (TP53, MED1, CCNB1 and EGF) were closely related to the survival of children with Wilms tumor. A 3-gene prognostic signature was constructed through LASSO regression analysis, and the patients stratified into with high- and low-risk groups based on this signature had significantly different survival outcomes (HR=1.814, log-rank P=0.002). The AUCs of the 3-, 5-and 7-year survival ROC curves of this model were all greater than 0.7. The overall mutations in the key hub genes or the individual mutations in TP53/CCNB1 were strongly correlated with a lower survival rates, and a high TP53 expression was correlated with a poor immunotherapy efficacy. RT-qPCR confirmed that the key hub genes had significant differential expressions in Wilms tumor tissues and cells. Conclusion TP53 gene plays an important role in the Wilms tumor and may potentially serve as a new immunotherapeutic biomarker as well as a therapeutic target.

Objective To identify the key genes differentially expressed in Wilms tumor and analyze their potential impacts on prognosis and immune responses of the patients. Methods High-throughput RNA sequencing was used to identify the differentially expressed mRNAs in clinical samples of Wilms tumor and paired normal tissues, and their biological functions were analyzed using GO, KEGG and GSEA enrichment analyses. The hub genes were identified using STRING database, based on which a prognostic model was constructed using LASSO regression. The mutations of the key hub genes were analyzed and their impacts on immunotherapy efficacy was predicted using the cBioPortal platform. RT-qPCR was used to verify the differential expressions of the key hub genes in Wilms tumor. Results Of the 1612 differentially expressed genes identified in Wilms tumor, 1030 were up-regulated and 582 were down-regulated, involving mainly cell cycle processes and immune responses. Ten hub genes were identified, among which 4 genes (TP53, MED1, CCNB1 and EGF) were closely related to the survival of children with Wilms tumor. A 3-gene prognostic signature was constructed through LASSO regression analysis, and the patients stratified into with high- and low-risk groups based on this signature had significantly different survival outcomes (HR=1.814, log-rank P=0.002). The AUCs of the 3-, 5-and 7-year survival ROC curves of this model were all greater than 0.7. The overall mutations in the key hub genes or the individual mutations in TP53/CCNB1 were strongly correlated with a lower survival rates, and a high TP53 expression was correlated with a poor immunotherapy efficacy. RT-qPCR confirmed that the key hub genes had significant differential expressions in Wilms tumor tissues and cells. Conclusion TP53 gene plays an important role in the Wilms tumor and may potentially serve as a new immunotherapeutic biomarker as well as a therapeutic target.

Objective To analyze the main active components and potential molecular mechanism of Sophora flavescens against breast cancer based on network pharmacology and molecular docking. Methods The chemical constituents were collected and screened by TCMSP, ETCM database and literature review. The targets of active ingredients were predicted by Swiss Target Prediction database. Breast cancer-related targets were collected by GeneCards, TTD, Drugbank and OMIM. The anti-breast cancer targets of Sophora flavescens were screened by Venny 2.1.0 software. Cytoscape software was used to construct the network diagram of Sophora flavescens-key active ingredients-targets. STRING database was used to analyze the common targets, and PPI network diagram was constructed. GO function enrichment analysis and KEGG pathway enrichment analysis of key target proteins were performed by DAVID database and Hiplot online platform. Schrodinger software was used to calculate the molecular docking between the active ingredients and targets. Molecular biological methods were used to verify the key targets. Results A total of 36 active components with clear structures were screened from Sophora flavescens. 70 anti-breast cancer targets of Sophora flavescens were screened out. 12 core targets including EGFR, AKT1, ESR1, SRC, CYP19A1, AR and ABCB1 participate in endocrine resistance, EGFR tyrosine kinase inhibitors and estrogen signaling pathways in breast cancer. Moreover, the docking score between the core component and the key target AR is the highest. In vitro experiments showed that the extract of Sophora flavescens can inhibit the proliferation of breast cancer cells, induce cell apoptosis and up-regulate AR protein expression. Conclusion It was revealed that Sophora flavescens plays an anti-breast cancer role by regulating complex biological processes through multiple components acting on multiple targets and signaling pathways. The upregulation of AR protein by Sophora flavescens may become a new therapeutic strategy for the treatment of breast cancer.

Objective: To explore the effect of C1q / tumor necrosis factor-related protein 9 ( CTRP9 ) on the expression of genes and proteins related to lipid metabolism of brown adipose tissue (BAT) in mice after cold stimulation． Methods : C57BL /6J male mice were injected with adenovirus Ad-GFP (control group) or Ad-CTRP9 ( experience group) into the scapular region and kept for 7 days．After cold stimulation at 4 ℃ for 10 hours，the expression levels of BAT marker genes and proteins were detected by real time PCR and Western blot. Results: Overexpression of CTRP9 induced by cold stimulation significantly increased the mRNA level of iodothyronine deiodinase 2 (Dio2) in BAT (P＜0. 01) ．Additionally，there was no significant difference in the expression of BAT marker genes ( UCP-1，PGC-1 α , PRDM16 and ARβ3) ，and liposynthesis and lipolysis related genes (PPARγ , HSL and ATGL) ．Uncoupling protein 1 (UCP-1) protein expression was upregualted in Ad-CTRP9 compared to the Ad-GFP control group ，while the expression of lipolysis related protein adipose triglyceride lipase ( ATGL) decreased significantly (P＜0. 05) ． Conclusion In cold environment，overexpression of CTRP9 promotes the accumulation of UCP-1 protein in BAT，upregulates the expression of thyroid hormone signal related gene Dio2，and inhibits triglyceride hydrolysis to maintain a constant body temperature.

Objective:To explore the correlation between system thinking ability and safety behavior of nurses in Emergency Department.Methods:From August 2021 to February 2022, convenience sampling was used to select nurses in Emergency Department from three ClassⅢ hospitals as the survey subject. Three rounds of questionnaire surveys were conducted using the Systems Thinking Scale and the Nurse Safety Behavior Questionnaire, with a 3-month interval among each round. The measurement time for each round was marked as T1, T2, and T3, respectively. AMOS 23.0 software was used for cross-lagged analysis to explore the correlation between Emergency Department nurses' system thinking ability and safety behavior. A total of 257 nurses in Emergency Department participated in the three rounds of questionnaire surveys.Results:The results of cross-lagged analysis showed that T1 system thinking ability could predict T2 safety behavior and T2 system thinking ability (β=0.26, 0.58, P<0.01). T1 safety behavior could predict T2 safety behavior and T2 system thinking ability (β=0.54, 0.17, P<0.01). T2 system thinking ability could predict T3 safety behavior and T3 system thinking ability (β=0.31, 0.59, P<0.01). T2 safety behavior could predict T3 safety behavior (β=0.48, P<0.01), but it could not predict T3 system thinking ability (β=0.04, P>0.05) . Conclusions:The system thinking ability and safety behavior of nurses in Emergency Department interact with each other, and the correlation develops dynamically. We should strengthen the cultivation of system thinking ability among nurses in Emergency Department to improve their safe behavior.

BACKGROUND: Although newly formed constructs of feasible pressure-preadjusted bone marrow mesenchymal stem cells (BMSCs) and platelet-rich fibrin (PRF) showed biomechanical flexibility and superior capacity for cartilage regeneration, it is still not very clear how BMSCs and seed cells feel mechanical stimuli and convert them into biological signals, and the difference in signal transduction underlying mechanical and chemical cues is also unclear. METHODS: To determine whether mechanical stimulation (hydrostatic pressure) and chemical cues (platelet-rich fibrin, PRF) activate canonical or noncanonical Wnt signaling in BMSCs, BMSCs cocultured with PRF were subjected to hydrostatic pressure loading, and the activation of the Wnt signaling molecules and expression of cartilage-associated proteins and genes were determined by western blotting and polymerase chain reaction (PCR). Inhibitors of canonical or noncanonical Wnt signaling, XVX-939 or L690,330, were adopted to investigate the role of Wnt signaling molecules in mechanically promoted chondrogenic differentiation of BMSCs. RESULTS: Hydrostatic pressure of 120 kPa activated both Wnt/b-catenin signaling and Wnt/Ca2+ signaling, with the the maximum promotion effect at 60 min. PRF exerted no synergistic effect on Wnt/b-catenin signaling activation. However, the growth factors released by PRF might reverse the promotion effects of pressure on Wnt/Ca2+ signaling. Real-time PCR and Western blotting results showed that pressure could activate the expression of Col-II, Sox9, and aggrecan in BMSCs cocultured with PRF. Blocking experiment found a positive role of Wnt/b-catenin signaling, and a negative role of Wnt/ Ca2+ signaling in chondrogenic differentiation of the BMSCs. Mutual inhibition exists between canonical and noncanonical Wnt signaling in BMSCs under pressure. CONCLUSION Wnt signaling participates in the pressure-promoted chondrogenesis of the BMSCs co-cultured with PRF, with canonical and noncanonical pathways playing distinct roles during the process.

Understanding the regulatory networks for germ cell fate specification is necessary to developing strategies for improving the efficiency of germ cell production in vitro. In this study, we developed a coupled screening strategy that took advantage of an arrayed bi-molecular fluorescence complementation (BiFC) platform for protein-protein interaction screens and epiblast-like cell (EpiLC)-induction assays using reporter mouse embryonic stem cells (mESCs). Investigation of candidate interaction partners of core human pluripotent factors OCT4, NANOG, KLF4 and SOX2 in EpiLC differentiation assays identified novel primordial germ cell (PGC)-inducing factors including BEN-domain (BEND/Bend) family members. Through RNA-seq, ChIP-seq, and ATAC-seq analyses, we showed that Bend5 worked together with Bend4 and helped mark chromatin boundaries to promote EpiLC induction in vitro. Our findings suggest that BEND/Bend proteins represent a new family of transcriptional modulators and chromatin boundary factors that participate in gene expression regulation during early germline development.
Animals ; Cell Differentiation/genetics* ; Chromatin/metabolism* ; Embryonic Stem Cells ; Germ Cells/metabolism* ; Germ Layers/metabolism* ; Mice