Objective To investigate changes in the microRNA expression profiles of apoptotic vesicles(apoVs)derived from bone marrow mesenchymal stem cells(BMSCs)under a simulated negative pressure environment,and to provide a theoretical basis for understanding the mechanism by which mechanical stress microenvironments influence the progression of osteoarthritis.Methods A negative pressure cellular environment was established using a pressure-loading system.Cell viability and apoptosis were assessed via the CCK-8 assay,Western blotting,and Annexin V-FITC/PI double staining.ApoVs were isolated by differential centrifugation and characterized using transmission electron microscopy(TEM),nanoparticle tracking analysis(NTA),and Western blotting.Small RNA sequencing was performed using the HiSeq Single-End mode,and differential expression analysis of microRNAs was conducted using DESeq to screen for differentially expressed microRNAs.The differentially screened microRNAs were validated by real-time quantitative PCR.After treating BMSCs with inhibitors of these differentially expressed microRNAs,the effects of the screened microRNAs on BMSCs were detected.Results Compared to apoVs generated by BMSCs under STS chemical treatment,those produced under a-40 kPa pressure environment showed significantly upregulated miR-183-5p and downregulated miR-3473.GO and KEGG enrichment analyses revealed that these differentially expressed microRNAs affected cell activity and inflammatory responses through multiple signaling pathways.Inhibition of miR-183-5p and miR-3473 expression reduced the proliferative activity of BMSCs.After inhibiting miR-183-5p expression,the levels of inflammatory factors increased.Inhibition of miR-3473 expression did not alter the IL-6 expression level,but significantly increased the TNFα expression level.Conclusions MicroRNAs specifically expressed in BMSC-derived apoVs under negative pressure stimulation may act as critical mechanical signaling mediators,regulating inflammatory response processes to participate in the pathogenesis and progression of arthritis.

Objective To investigate the anxiety and its related factors in medical staff during field training,so as to provide evidence for psychological education before field training.Methods A total of 81 medical staff who participated in field training were recruited in this cross-sectional study as research objects.The Hamilton Anxiety Scale(HAMA)and self-compiled questionnaires of related factors were used to measure the general states of the staff.Univariate analysis was used to screen relevant variables that affect anxiety,and then a logistic regression model was used for multivarate analysis to determine independent influencing factors.Results The detection rate of anxiety was 14.81%in the research objects.There were significant differences in the degree of anxiety among the medical staff with different variables such as gender,the number of exercises attended,childlessness,parental physical condition,tolerable training time,pastime activities,and personal title(P<0.05).Multivariate regression analysis indicated that gender and parental physical status were independent factors affecting anxiety(P<0.05).Conclusion During field training,a part of medical staff has anxiety,which may affect by multi-factors such as personal,family,and training environment.Therefore,psychological education and counseling before field training should be performed from multiple aspects.

Objective To investigate changes in the microRNA expression profiles of apoptotic vesicles(apoVs)derived from bone marrow mesenchymal stem cells(BMSCs)under a simulated negative pressure environment,and to provide a theoretical basis for understanding the mechanism by which mechanical stress microenvironments influence the progression of osteoarthritis.Methods A negative pressure cellular environment was established using a pressure-loading system.Cell viability and apoptosis were assessed via the CCK-8 assay,Western blotting,and Annexin V-FITC/PI double staining.ApoVs were isolated by differential centrifugation and characterized using transmission electron microscopy(TEM),nanoparticle tracking analysis(NTA),and Western blotting.Small RNA sequencing was performed using the HiSeq Single-End mode,and differential expression analysis of microRNAs was conducted using DESeq to screen for differentially expressed microRNAs.The differentially screened microRNAs were validated by real-time quantitative PCR.After treating BMSCs with inhibitors of these differentially expressed microRNAs,the effects of the screened microRNAs on BMSCs were detected.Results Compared to apoVs generated by BMSCs under STS chemical treatment,those produced under a-40 kPa pressure environment showed significantly upregulated miR-183-5p and downregulated miR-3473.GO and KEGG enrichment analyses revealed that these differentially expressed microRNAs affected cell activity and inflammatory responses through multiple signaling pathways.Inhibition of miR-183-5p and miR-3473 expression reduced the proliferative activity of BMSCs.After inhibiting miR-183-5p expression,the levels of inflammatory factors increased.Inhibition of miR-3473 expression did not alter the IL-6 expression level,but significantly increased the TNFα expression level.Conclusions MicroRNAs specifically expressed in BMSC-derived apoVs under negative pressure stimulation may act as critical mechanical signaling mediators,regulating inflammatory response processes to participate in the pathogenesis and progression of arthritis.

Ligand-gated ion channels transduce chemical signals into ionic currents while being bonded with specific ligands,thereby facilitating signal transduction and intercellular communication.These channels have become critical therapeutic targets for a variety of diseases.Ginsenosides,triter-penoid saponins primarily derived from herbs of the Panax genus,are known for their broad range of biological activities.Ginsenosides and their metabolites can regulate the functions of various ligand-gated ion channels,including nicotinic acetylcholine receptors,5-hydroxytryptamine type-3 receptors,γ-aminobu-tyric acid type-A receptors,glycine receptors,ionotropic glutamate receptors,ATP-gated P2X chan-nels,and transient receptor potential channels.They play important roles in anti-inflammation,analge-sics,immune regulation,anti-epilepsy,cognitive enhancement and neuroprotection.Further investiga-tion into the mechanisms by which ginsenosides regulate these channels will provide important evidence for their clinical applications and the development of new drugs.

Objective To evaluate the cost-utility of tiotropium/olodaterol in treating Chinese patients with moderate to very severe chronic obstructive pulmonary disease(COPD)and to provide references for selecting more economical inhaled preparations in clinical practice.Methods A four-state lifetime Markov model was established with a 3-month cycle.The health outcomes included life years and quality-adjusted life years.Costs,including direct medical costs,were calculated from the perspective of the Chinese health system.Discontinuation rates were derived from the discontinuation curve using GetData Graph Digitizer.The main output indicator of the model was the incremental cost-utility ratio,which was calculated from the queue simulation results to judge the economy of tiotropium bromide/odataterol.The scenario analysis and sensitivity analyses were carried out to detect the robustness of the base case result.Results Compared with tiotropium bromide,the patient treated with tiotropium bromide/odataterol gained an additional 0.0846 life years,an additional cost of ￥3 201.50,and additional 0.029 6 QALY.The incremental cost-utility ratio was 108 140.11 yuan/QALY,lower than the willingness-to-pay threshold of three times China's per capita GDP in 2021.The costs of tiotropium bromide and tiotropium bromide/odataterol had the greatest impact on the result in the one-way sensitivity analysis.93.8%of the Chinese COPD population was willing to pay for tiotropium bromide/odataterol under the threshold in the probability sensitivity analysis.Conclusion Tiotropium/olodaterol is a cost-effective alternative compared to tiotropium for patients with moderate to very severe COPD in China and the results were robust in the sensitivity analyses.

This study aimed to explore the roles of microRNAs (miRNAs) in the post-transcriptional regulation of cocaine- and amphetamine-regulated transcript (CART) peptide in the bovine hypothalamus and to screen key regulatory miRNAs. Targetscan was used to predict the potential miRNAs binding to CART 3' untranslated regions (3'UTR). Bioinformatics analysis predicted 7 miRNA binding sites in the bovine CART 3'UTR, which were bta-miR-377, bta-miR-331-3p, bta-miR-491, bta-miR-493, bta-miR-758, bta-miR-877, and bta-miR-381, respectively. Reverse transcription-PCR (RT-PCR) was carried out to determine the endogenous expression of CART and target miRNAs in the bovine hypothalamus. All the 7 target miRNAs and CART were endogenously expressed in the bovine hypothalamus. The dual-luciferase reporter gene assay was employed to detect the targeted binding relationship between CART 3'UTR and target miRNAs obtained from bioinformatics analysis. The dual-luciferase reporter gene assay confirmed that the 3'UTR of CART had a targeted binding relationship with the 7 target miRNAs. Cell experiments were conducted to examine the effects of target miRNAs on the messenger RNA (mRNA) and protein levels of exogenous CART and screen for key regulatory miRNAs. The results of cell experiments showed that the 7 miRNAs downregulated the mRNA level of CART, with bta-miR-491 demonstrating the strongest downregulating effect. Bta-miR-377, bta-miR-331-3p, bta-miR-491, bta-miR-493, and bta-miR-381 downregulated the protein level of CART, with bta-miR-381 exerting the strongest downregulating effect. Animal experiments were conducted to explore the effects of key regulatory miRNAs on the mRNA and protein levels of CART in the hypothalamus and the CART concentration in the serum. The results from animal experiments showed that miR-491 and miR-381 regulated the endogenous expression of CART in the hypothalamus and the concentration in the serum by binding to the CART 3'UTR. These results suggest that miR-491 and miR-381 are the main miRNAs regulating CART expression in the bovine hypothalamus, which can affect serum CART concentration by modulating endogenous CART expression.
Animals ; MicroRNAs/metabolism* ; Cattle ; Hypothalamus/metabolism* ; 3' Untranslated Regions/genetics* ; Nerve Tissue Proteins/metabolism* ; Gene Expression Regulation ; Binding Sites ; Base Sequence ; Computational Biology/methods* ; Cocaine- and Amphetamine-Regulated Transcript Protein

Intestinal fibrosis in Crohn′s disease (CD) presents a significant clinical challenge, imposing a substantial burden on patients, their families and society. This article explores the histopathological and pathophysiological changes associated with intestinal fibrosis, providing a detailed summary of the related phenomena and processes. Additionally, we discuss the methods for detecting and assessing intestinal fibrosis, including radiological examinations, histopathological investigations, and the application of serum biomarkers. Finally, we delve into the therapeutic strategies for intestinal fibrosis in CD and the challenges faced in this realm, offering vital insights for further research and treatment of the disease.

AIM:To analyze and screen the risk factors for acute kidney injury(AKI)following heart transplantation(HT),and to establish a dynamic no-mograms prediction model to forecast early AKI af-ter HT.METHODS:A retrospective analysis was con-ducted on clinical data from HT recipients at Nan-jing First Hospital from October 2012 to June 2024.Patients were divided into AKI and non-AKI groups based on AKI occurrence within 7 days post-sur-gery,with a 8:2 ratio for training and testing sets.Lasso regression and multivariable logistic regres-sion were used to select influencing factors.A dy-namic nomogram model was visualized using R.In-ternal validation was performed using 1 000 boot-strap samples.Model accuracy and discrimination were evaluated using the area under the receiver operating characteristic curve(AUC-ROC),calibra-tion curves,and the Hosmer-Lemeshow goodness-of-fit test.The nomogram model was compared with the Cleveland score.RESULTS:The results of a multivariable logistic regression indicate that a his-tory of atrial fibrillation(OR=9.647,95％CI=1.961-47.470),vasoactive inotropic score(OR=1.094,95％CI=1.012-1.183),intraoperative transfusion of red blood cells or plasma(OR=10.200,95％CI=1.727-60.238),postoperative central venous pressure(OR=1.548,95％CI=1.186-2.021),and postoperative use of vancomycin(OR=25.082,95％CI=2.122-296.417)are independent risk factors for HT-AKI.The dynamic nomogram model achieved an AUC of 0.842(95％CI:0.676-0.971)in the test set,with a calibration plot showing a slope close to 1 and a Brier score of 0.173.The Hosmer-Lemeshow good-ness-of-fit test(x2=5.658,P=0.685)suggests good predictive performance of the model.Moreover,this model demonstrates superior discriminative ability compared to the Cleveland score.CONCLU-SION:This study identified preoperative,intraoper-ative,and postoperative risk factors influencing the occurrence of HT-AKI.The developed dynamic no-mogram model accurately identifies high-risk indi-viduals for early HT-AKI and is convenient for clini-cal use.

Intestinal fibrosis in Crohn′s disease (CD) presents a significant clinical challenge, imposing a substantial burden on patients, their families and society. This article explores the histopathological and pathophysiological changes associated with intestinal fibrosis, providing a detailed summary of the related phenomena and processes. Additionally, we discuss the methods for detecting and assessing intestinal fibrosis, including radiological examinations, histopathological investigations, and the application of serum biomarkers. Finally, we delve into the therapeutic strategies for intestinal fibrosis in CD and the challenges faced in this realm, offering vital insights for further research and treatment of the disease.

AIM:To analyze and screen the risk factors for acute kidney injury(AKI)following heart transplantation(HT),and to establish a dynamic no-mograms prediction model to forecast early AKI af-ter HT.METHODS:A retrospective analysis was con-ducted on clinical data from HT recipients at Nan-jing First Hospital from October 2012 to June 2024.Patients were divided into AKI and non-AKI groups based on AKI occurrence within 7 days post-sur-gery,with a 8:2 ratio for training and testing sets.Lasso regression and multivariable logistic regres-sion were used to select influencing factors.A dy-namic nomogram model was visualized using R.In-ternal validation was performed using 1 000 boot-strap samples.Model accuracy and discrimination were evaluated using the area under the receiver operating characteristic curve(AUC-ROC),calibra-tion curves,and the Hosmer-Lemeshow goodness-of-fit test.The nomogram model was compared with the Cleveland score.RESULTS:The results of a multivariable logistic regression indicate that a his-tory of atrial fibrillation(OR=9.647,95％CI=1.961-47.470),vasoactive inotropic score(OR=1.094,95％CI=1.012-1.183),intraoperative transfusion of red blood cells or plasma(OR=10.200,95％CI=1.727-60.238),postoperative central venous pressure(OR=1.548,95％CI=1.186-2.021),and postoperative use of vancomycin(OR=25.082,95％CI=2.122-296.417)are independent risk factors for HT-AKI.The dynamic nomogram model achieved an AUC of 0.842(95％CI:0.676-0.971)in the test set,with a calibration plot showing a slope close to 1 and a Brier score of 0.173.The Hosmer-Lemeshow good-ness-of-fit test(x2=5.658,P=0.685)suggests good predictive performance of the model.Moreover,this model demonstrates superior discriminative ability compared to the Cleveland score.CONCLU-SION:This study identified preoperative,intraoper-ative,and postoperative risk factors influencing the occurrence of HT-AKI.The developed dynamic no-mogram model accurately identifies high-risk indi-viduals for early HT-AKI and is convenient for clini-cal use.