OBJECTIVES: Osteoarthritis (OA) and sarcopenia are significant health concerns in the elderly, substantially impacting their daily activities and quality of life. However, the relationship between them remains poorly understood. This study aims to uncover common biomarkers and pathways associated with both OA and sarcopenia. METHODS: Gene expression profiles related to OA and sarcopenia were retrieved from the Gene Expression Omnibus (GEO) database. Differentially expressed genes (DEGs) between disease and control groups were identified using R software. Common DEGs were extracted via Venn diagram analysis. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were conducted to identify biological processes and pathways associated with shared DEGs. Protein-protein interaction (PPI) networks were constructed, and candidate hub genes were ranked using the maximal clique centrality (MCC) algorithm. Further validation of hub gene expression was performed using 2 independent datasets. Receiver operating characteristic (ROC) curve analysis was used to evaluate the predictive value of key genes for OA and sarcopenia. Mouse models of OA and sarcopenia were established. Hematoxylin-eosin and Safranin O/Fast Green staining were used to validate the OA model. The sarcopenia model was validated via rotarod testing and quadriceps muscle mass measurement. Real-time reverse transcription PCR (real-time RT-PCR) was employed to assess the mRNA expression levels of candidate key genes in both models. Gene set enrichment analysis (GSEA) was conducted to identify pathways associated with the selected shared key genes in both diseases. RESULTS: A total of 89 common DEGs were identified in the gene expression profiles of OA and sarcopenia, including 76 upregulated and 13 downregulated genes. These 89 DEGs were significantly enriched in protein digestion and absorption, the PI3K-Akt signaling pathway, and extracellular matrix-receptor interaction. PPI network analysis and MCC algorithm analysis of the 89 common DEGs identified the top 17 candidate hub genes. Based on the differential expression analysis of these 17 candidate hub genes in the validation datasets, AEBP1 and COL8A2 were ultimately selected as the common key genes for both diseases, both of which showed a significant upregulation trend in the disease groups (all P<0.05). The value of area under the curve (AUC) for AEBP1 and COL8A2 in the OA and sarcopenia datasets were all greater than 0.7, indicating that both genes have potential value in predicting OA and sarcopenia. Real-time RT-PCR results showed that the mRNA expression levels of AEBP1 and COL8A2 were significantly upregulated in the disease groups (all P<0.05), consistent with the results observed in the bioinformatics analysis. GSEA revealed that AEBP1 and COL8A2 were closely related to extracellular matrix-receptor interaction, ribosome, and oxidative phosphorylation in OA and sarcopenia. CONCLUSIONS AEBP1 and COL8A2 have the potential to serve as common biomarkers for OA and sarcopenia. The extracellular matrix-receptor interaction pathway may represent a potential target for the prevention and treatment of both OA and sarcopenia.
Sarcopenia/genetics* ; Osteoarthritis/genetics* ; Computational Biology/methods* ; Humans ; Protein Interaction Maps/genetics* ; Animals ; Mice ; Gene Expression Profiling ; Gene Ontology ; Transcriptome ; Male ; Signal Transduction/genetics* ; Gene Regulatory Networks

OBJECTIVES: Osteoarthritis (OA) is one of the most common chronic degenerative diseases, with chondrocyte apoptosis and extracellular matrix (ECM) degradation as the major pathological changes. The mechanical stimulation can attenuate chondrocyte apoptosis and promote ECM synthesis, but the underlying molecular mechanisms remain unclear. This study aims to investigate the role of primary cilia (PC) in mediating the effects of mechanical stimulation on OA progression. METHODS: In vivo, conditional knockout mice lacking intraflagellar transport 88 (IFT88^flox/flox IFT88 knockout; i.e., primary cilia-deficient mice) were generated, with wild-type mice as controls. OA models were established via anterior cruciate ligament transection combined with destabilization of the medial meniscus, followed by treadmill exercise intervention. OA progression was evaluated by hematoxylin-eosin staining, safranin O-fast green staining, and immunohistochemistry; apoptosis was assessed by TUNEL staining; and limb function by rotarod testing. In vitro, primary articular chondrocytes were isolated from mice and transfected with lentiviral vectors to suppress IFT88 expression, thereby constructing a primary cilia-deficient cell model. Interleukin-1β (IL-1β) was used to induce an inflammatory environment, while cyclic tensile strain (CTS) was applied via a cell stretcher to mimic mechanical loading on chondrocytes. Immunofluorescence and Western blotting were used to detect the protein expression levels of type II collagen α1 chain (COL2A1), primary cilia, IFT88, and caspase-12; reverse transcription polymerase chain reaction was performed to assess COL2A1 mRNA levels; and flow cytometry was used to evaluate apoptosis. RESULTS: In vivo, treadmill exercise significantly reduced Osteoarthritis Research Society International (OARSI) scores and apoptotic cell rates, and improved balance ability in wild-type OA mice, whereas IFT88-deficient OA mice showed no significant improvement. In vitro, CTS inhibited IL-1β-induced ECM degradation and apoptosis in primary chondrocytes; however, this protective effect was abolished in cells with suppressed primary cilia expression. CONCLUSIONS Mechanical stimulation delays OA progression by mediating signal transduction through primary cilia, thereby inhibiting cartilage degeneration and chondrocyte apoptosis.
Animals ; Chondrocytes/cytology* ; Apoptosis/physiology* ; Mice ; Cilia/metabolism* ; Osteoarthritis/pathology* ; Extracellular Matrix/metabolism* ; Mice, Knockout ; Disease Progression ; Interleukin-1beta ; Male ; Cells, Cultured

The control strategy of rehabilitation robots should not only adapt to patients with different levels of motor function but also encourage patients to participate voluntarily in rehabilitation training.However,existing control strategies usually consider only one of these aspects.This study proposes a voluntary and adaptive control strategy that solves both questions.Firstly,the controller switched to the corresponding working modes(including challenge,free,assistant,and robot-dominant modes)based on the trajectory tracking error of human-robot cooperative movement.To encourage patient participation,a musculoskeletal model was used to estimate the patient's active torque.The robot's output torque was designed as the product of the active torque and a coefficient,with the coefficient adaptively changing according to the working mode.Experiments were conducted on two healthy subjects and four hemiplegic patients using an ankle rehabilitation robot.The results showed that this controller not only provided adaptive the robot's output torque based on the movement performance of patients but also encouraged patients to complete movement tasks themselves.Therefore,the control strategy has high application value in the field of rehabilitation.

ObjectiveTo explore the role of saikosaponin D （SSD） targeting signal transducer and activator of transcription 3 （STAT3） in inducing apoptosis of bladder cancer cells by computer-aided drug design and experimental verification. MethodThe druggability and biotoxicity of SSD were explored by Bayesian classifier modeling. The information about SSD， the active ingredient of Bupleuri Radix， was searched against the Traditional Chinese Medicine Systematic Pharmacology Database and Analysis Platform （TCMSP）. The targets of SSD were predicted by PubChem， TCMSP， a Bioinformatics Analysis Tool for Molecular mechANism of Traditional Chinese Medicine （BATMAN-TCM）， Coremine， an Encyclopedia of Traditional Chinese Medicine （ETCM）， and SwissTargetPrediction. GeneCards， Therapeutic Target Database （TTD）， and Online Mendelian Inheritance in Man （OMIM） were employed to predict the potential therapeutic targets of bladder cancer. Then， the common targets shared by SSD and bladder cancer were selected for Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analyses. Molecular docking was adopted to explore the binding affinity and structural stability of SSD with target proteins. Cytoscape 3.9.1 was used to construct the STAT3-drug regulatory network and STAT3-apoptosis regulatory network. UM-UC-3 cells were treated with 0， 5， 10， 15 μmol·L^-1 SSD for 24 h. Then， flow cytometry was used to detect the apoptosis of bladder cancer cells， and Western blot was employed to determine the protein levels of B-cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， Bcl-2-associated death promoter （Bad）， STAT3， and phosphorylation （p）-STAT3. ResultBayesian classifier modeling and molecular docking showed that SSD had low biotoxicity and bound well to the target protein STAT3 to form a stable protein-ligand complex. There were 282 common targets between bladder cancer and SSD， among which STAT3 was the most central target. The GO enrichment analysis showed that the potential core therapeutic targets involved 3 036 biological processes， 82 cellular components， and 171 molecular functions. The KEGG enrichment analysis showed that the potential core targets were mainly related to the C-type lectin receptor signaling pathway， Toll-like receptor signaling pathway， and cell apoptosis pathway. The STAT3-drug regulatory network and STAT3-apoptosis regulatory network showed that 29 drugs interacted with STAT3， and 27 apoptosis-related genes had a strong correlation with STAT3. Flow cytometry showed that the apoptosis rate increased with the increase in SSD concentration （P<0.05）. Western blotting results showed that SSD down-regulated the protein levels of p-STAT3 and Bcl-2 and up-regulated the protein levels of Bax and Bad in a concentration-dependent manner （P<0.05）. ConclusionSSD has good druggability and low biotoxicity. It may promote the apoptosis of bladder cancer cells by targeting STAT3.

Objective:To understand the problems existing in the curriculum design of food hygiene and nutrition for undergraduates, and put forward countermeasures to improve the quality of talents cultivation and promote professional development.Methods:Self-made questionnaires were distributed by convenience sampling method, through online and offline ways, and the graduates' opinions on the curriculum design of their school. Quantitative data were analyzed by SPSS 26.0, and qualitative data were analyzed by NVivo 11.0.Results:A total of 258 valid questionnaires were recovered, and the average score of graduates' satisfaction with teaching quality was 2.42±1.02. Graduates had different opinions on the importance of special courses, and 150 students (58.14%) were satisfied with major courses. Some graduates thought the time of special courses was unreasonable and there were few major courses. Besides, 141 graduates (54.65%) were satisfied with the clinical probation in their graduation colleges. Although graduates believed that innovation ability was important, innovative education courses were scarce at present.Conclusions:Graduates are less satisfied with the setting of undergraduate curriculum in food hygiene and nutrition. It is suggested to optimize professional courses setting, improve experimental facilities and equipment, strengthen the management of practice bases, and pay attention to the cultivation of students' innovation and entrepreneurship ability.

Objective:To evaluate the quality of test papers and to analyze students'mastery of knowledge through the analysis of test papers,so as to provide reference for the reform of test proposition and correction teaching.Methods:Using paper analysis software(Ver 2.0),the paper quality,the students'scores and the answers to the questions of the final exam paper of Physiology of grade 2021 students majoring in nursing were analyzed.Results:The composition of the test paper was consistent with the requirements of the teaching programme,the difficulty of subjective and objective test questions was moderate,the differentiation of subjective test questions was good,the differentiation of objective test questions was general,and the reliability and validity were good.The overall performance was basically normal distribution.The full score ratio of objective test questions was higher than that of subjective test questions,and the zero score test questions were mostly concentrated in chapter 10 and chapter 4.Conclusion:Test paper analysis can feedback the problems and shortcomings of test paper proposition and teaching process,promote the quality of test paper and teaching model innovation,and improve the quality of teaching.

Objective:To investigate the short-term efficacy and safety of Donafenib as postoperative adjuvant therapy for patients with high risk of recurrence after radical resection of hepatocellular carcinoma (HCC).Methods:The propensity score matching (PSM) and retrospective cohort study was conducted. The clinicopathological data of 157 HCC patients with high risk of recurrence after radical resection who were admitted to 6 medical centers, including The First Affiliated Hospital of Nanjing Medical University et al, from June 2021 to February 2023 were collected. There were 128 males and 29 females, aged (59±10)years. Of 157 patients, 101 cases undergoing Donafenib as postoperative adjuvant therapy were divided into the the Donafenib group, and 56 cases under-going no systemic postoperative adjuvant therapy were divided into the control group. Observation indicators: (1) PSM and comparison of general data of patients between the two groups after matching; (2) postoperative treatment; (3) follow-up and survival of patients; (4) analysis of risk factors affecting recurrence-free survival of patients. PSM was done based on the principle of optimal perfect matching, with the clamp value of 0.5, and the Donafenib group and the control group were matched at a ratio of 1.25∶1. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the t test. Measurement data with skewed distribution were represented as M(range). Count data were described as absolute numbers and/or percentages, and comparison between groups was conducted using the chi-square test. Comparison of ordinal data between groups was conducted using the Kruskal-Wallis H test. The Kaplan-Meier method was used to calculate survival rates and draw survival curves, and the Log-Rank test was used for survival analysis. The COX proportional hazard model was used for univariate and multivariate analyses. Results:(1) PSM and comparison of general data of patients between the two groups after matching. Of 157 patients, 126 cases were successfully matched, including 70 cases in the Donafenib group and 56 cases in the control group, respectively. The elimination of tumor number confounding bias ensured comparability between the two groups after PSM. (2) Postoperative treatment. After PSM, of 70 patients in the Donafenib group, there were 23 cases receiving Donafenib monotherapy, 26 cases combined with transcatheter arterial chemoembolization (TACE), 14 cases combined with immunotherapy, and 7 cases combined with TACE+immunotherapy. Of 56 patients in the control group, there were 37 cases receiving postoperative follow-up alone and 19 cases combined with TACE. (3) Follow-up and survival of patients. All 157 patients were followed up, and the follow-up time of the 101 patients in Donafenib group and the 56 patients in control group were 10.1(range, 6.3-14.6)months and 22.2(range, 15.1-25.5)months, respectively. During the follow-up period, 70 patients in the Donafenib group experienced treatment-related adverse reactions, inclu-ding 8 cases of grade 3 adverse reactions, 23 cases of grade 2 and 39 cases of grade 1 adverse reactions, respectively. After PSM, the postoperative 12-, 18-month recurrence-free survival rates were 83.7%, 83.7% in the 70 patients of Donafenib group and 67.8%, 58.9% in the 56 patients of control group, respectively, showing a significant difference in the postoperative recurrence-free survival time between the two groups ( hazard ratio=0.395, 95% confidence interval as 0.176-0.888, P<0.05). (4) Analysis of risk factors affecting recurrence free survival of patients. Results of multivariate ana-lysis showed that microvascular invasion, vascular thrombus, clinical stage as ⅢA were independent risk factors affecting recurrence-free survival in patients with high risk of recurrence after radical resection of HCC ( hazard ratio=2.181, 2.612, 2.612, 95% confidence interval as 1.028-4.629, 1.128-6.047, 1.128-6.047, P<0.05), Donafenib as postoperative adjuvant therapy was an independent protective factor affecting recurrence-free survival in patients with high risk of recurrence after radical resection of HCC ( hazard ratio=0.457, 95% confidence interval as 0.227-0.920, P<0.05). Results of further analysis showed that after PSM, there were significant differences in the postoperative recurrence-free survival time in patients with different clinical factors, including male, age ≥60 years, tumor diameter >5 cm, positive microvascular invasion, positive hepatitis B virus infection, alpha fetoprotein <200 μg/L, between the Donafenib group and the control group ( hazard ratio=0.283, 0.202, 0.174, 0.345, 0.273, 0.180, 95% confidence interval as 0.114-0.707, 0.044-0.937, 0.038-0.794, 0.128-0.929, 0.091-0.819, 0.052-0.620, P<0.05). Conclusion:Donafenib as postoperative adjuvant therapy can effectively reduce the short-term recurrence rate in patients with high risk of recurrence after radical resection of HCC, with good safety and tolerance.

Objective:To investigate the association of serum cholinesterase(ChE) with insulin resistance(IR) in patients with type 2 diabetes mellitus(T2DM).Methods:General information and laboratory results of 376 hospitalized patients with type 2 diabetes were collected from the Endocrinology Department of Xuzhou Medical University Affiliated Hospital from June 2020 to November 2022. Based on quartile levels of serum cholinesterase(ChE), the patients were divided into four groups: Q1 group(≤7 912 IU/L), Q2 group(7 913-9 083 IU/L), Q3 group(9 084-10 304 IU/L), and Q4 group(≥10 305 IU/L). The Homeostasis model was used to evaluate insulin resistance(HOMA-IR) for each group, and the correlation between ChE and HOMA-IR was analyzed. Results:As ChE levels increased, HOMA-IR levels significantly increased( P<0.05). Spearman correlation analysis showed that ChE was positively correlated with body mass index, diastolic blood pressure, high-sensitivity C-reactive protein, HbA 1C, total cholesterol, triglycerides, low density lipoprotein-cholesterol(LDL-C), aspartate aminotransferase, alanine aminotransferase(ALT), glutamyl transpeptidase, prealbumin, albumin, uric acid, and HOMA-IR( P<0.05). ChE was negatively correlated with age, duration of diabetes, and blood urea nitrogen( P<0.05). Multivariat stepwise linear regression analysis showed that disease duration, body mass index, HbA 1C, ALT, and ChE were independent influencing factors of HOMA-IR( P<0.01). Logistic regression analysis showed that compared to the Q1 group, the risks of developing IR in the Q4, Q3, and Q2 groups were 3.969(1.791-8.797), 2.100(1.059-4.164), and 2.026(1.071-3.833), respectively. Conclusion:Serum ChE is closely associated with IR in patients with T2DM.

Objective:To explore the role of 17β-estradiol(17β-E2) derived from the hippocampus and gonad in the cognitive memory during the reconsolidation period of fear memory in rats with post-traumatic stress disorder(PTSD).Methods:The 8-week-old clean grade female SD rats were used for this study. Single prolonged stress combined with context fear conditioning was used to prepare the rat PTSD model.(1) Gonad estradiol experiment: 50 female rats were randomly divided into blank control group, model group, sham ovariectomy group, ovariectomy group, and ovariectomy+ estradiol group by random number table method, with 10 rats in each group. The rats in model group, sham ovariectomy group, ovariectomy group, and ovariectomy+ estradiol group were established PTSD model. Rats in ovariectomy group and ovariectomy+ estradiol group underwent ovariectomy, while rats in sham ovariectomy group only were removed adipose tissue of the same mass around the ovaries. And the rats in ovariectomy+ estradiol group were injected with 17β-E2(1 mg/kg, once a day for 14 days) 7 days after ovariectomy.(2) Hippocampal estradiol experiment: 40 female rats were randomly divided into blank control group, model group, hippocampal solvent control group and hippocampal estradiol inhibitor group by random number table method, with 10 rats in each group.The rats in model group, hippocampal solvent control group and hippocampal estradiol inhibitor group were established PTSD model.Rats in hippocampal estradiol inhibitor group were given letrozole once(bilateral, 0.5 μL per side) during the fear memory consolidation period) and rats in the hippocampus solvent control group were injected once with dimethyl sulfoxide in the hippocampus(bilateral, 0.5 μL per side) . The open field test and elevated cross maze test were used to evaluate the anxiety level and autonomous exploration ability, the freeze test was used to evaluate the fear memory, the new object recognition test was used to evaluate the non-spatial memory, and ELISA was used to detect serum 17β-E2 level. SPSS 25.0 software was used for data analysis, and one-way ANOVA was used for inter group comparisons, LSD test was used for further pairwise comparisons.Results:(1)In the gonad estradiol experiment, there were statistical differences in various indicators among the five groups of rats in the open field test, elevated cross maze test, freeze test, and new object recognition test( F=20.200, 12.702, 7.514, 10.094, 7.899, 13.211, all P<0.05). The number of upright times, central area activity time, distance and frequency of entering the open arm, and cognitive index of the model group rats were all lower than those of the blank control group(all P<0.05), and the freezing time was higher than that of the blank control group( P<0.05). The number of upright positions((11.20±1.55) times), central area activity time((11.33±1.80) s), distance((1.49±0.26) m) and times((10.00±1.50) times) entering the open arm, the freezing time((92.20±6.07) s) and cognitive index((60.40±3.71)%) in ovariectomy+ estradiol group were all higher than those of ovariectomy group((4.90±0.65) times, (4.31±1.07) s, (0.49±0.06) m, (3.10±0.62) times, (60.30±5.28) s, (32.60±8.08)%)(all P<0.05). (2) In the hippocampal estradiol experiment, there were statistical differences in various indicators among the four groups in the open field test, elevated cross maze test, freeze test, and new object recognition test( F=40.831, 5.553, 9.087, 5.848, 7.657, 9.191, all P<0.05). The numbers of upright positions, distance and frequency of entering open arms of the model group rats were lower than those of the blank control group(all P<0.05). The number of upright positions((3.00 ± 0.39) times), distance of entering open arm((1.17±0.37) m), freezing time((46.70±3.57)s), and cognition index((29.60±2.70)%) in the hippocampal estradiol inhibitor group were all lower than those in the hippocampal solvent control group((10.10±1.40) times, (4.02±0.79) m, (93.70±9.73) s, (54.20±5.08)%)(all P<0.05). Conclusion:17β-E2 derived from the hippocampus and gonad both had ameliorative effect on cognitive memory and anxiety-like behavior in the reconsolidation period of fear memory in PTSD model rats.

Objective:To explore the application of PDCA (plan, do, check, act) cycle theory in the teaching of eight-year clinical medical students for the course of Laboratory Diagnostics.Methods:From September 2021 to November 2022, the students majoring in clinical medicine of eight-year system who were studying the course of Laboratory Diagnostics at Peking Union Medical College were selected as the research objects. The teaching reform of the course of Laboratory Diagnostics was carried out according to the four stages of PDCA cycle theory, and the teaching effect was evaluated by the final examination results and questionnaire survey scores of the students.Results:The score of theoretical examination of eight-year students in 2018 was (86.7±4.68) points, which was higher than that of students in 2017 [(83.3±3.89) points], and the difference was statistically significant ( P<0.05). The questionnaire survey results of the two groups of students showed that the rating of experimental course was higher than that of theoretical course ( P<0.05). After the teaching reform based on PDCA cycle theory, the questionnaire survey scores of theoretical course [(3.83±0.25) points vs (2.94±0.28) points] and experimental course [(4.13±0.09) points vs (3.32±0.12) points] in students of 2018 were higher than those of 2017 (all P<0.001). Conclusion:PDCA cycle theory provides new methods and ideas for teaching management, which helps to improve the performance of clinical medical students of eight-year system and their recognition of the course.