ObjectiveTo investigate the mechanism by which Gegen Qinliantang（GQT） improves skeletal muscle insulin resistance. MethodsThe db/m mice were used as the normal group， while db/db mice were assigned to a model group， low-dose （3.12 g·kg^-1）， medium-dose （6.24 g·kg^-1）， and high-dose （12.48 g·kg^-1） GQT groups， and a Western medicine group （semaglutide， 0.045 mg·kg^-1），n=6 in each group. All groups received corresponding interventions. Intraperitoneal glucose tolerance test （IPGTT）， intraperitoneal insulin tolerance test （IPITT）， and hematoxylin-eosin （HE） staining were used to evaluate insulin resistance and therapeutic efficacy. Serum lipid levels were measured using an automatic biochemical analyzer， and apoptosis in skeletal muscle was assessed via TUNEL assay. Transcriptome sequencing combined with gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） analyses was performed to identify differentially expressed genes （DEGs）. Real-time quantitative polymerase chain reaction （Real-time PCR） was used to validate gene expression. Molecular docking was applied to evaluate the binding patterns between active components of GQT and key regulatory genes to elucidate pharmacological mechanisms. ResultsCompared with the model group， the medium-dose and high-dose GQT groups showed significantly reduced fasting blood glucose （FBG） levels （P<0.01）. Triglycerides （TG）， total cholesterol （TC）， and low-density lipoprotein cholesterol （LDL-C） were markedly decreased （P<0.01）， while high-density lipoprotein cholesterol （HDL-C） was significantly increased （P<0.01）. IPGTT， IPITT， and HE staining demonstrated that GQT enhanced insulin sensitivity and restored skeletal muscle morphology. GQT also alleviated apoptosis in skeletal muscle tissue. Transcriptome analysis revealed that GQT primarily affected biological processes such as oxidative phosphorylation， metabolic pathways， cellular processes， and protein binding. Real-time PCR results showed that CBR2， CDK6， F830016B08Rik， IL-1β， Rab27b， and COLEC12 were key regulatory genes. Molecular docking demonstrated that CBR2， IL-1β， Rab27b， and COLEC12 formed stable binding with the main active components of GQT. The therapeutic effects of high- and medium-dose GQT were comparable to those of the semaglutide group. ConclusionGQT improves skeletal muscle insulin resistance， potentially by regulating apoptosis as part of its underlying biological mechanism.

ObjectiveTo investigate the effects of salidroside （SAL） on the impaired bioactivity of endothelial progenitor cells （EPCs） in atherosclerotic （As） mice and the potential mechanisms regarding AMP-activated protein kinase （AMPK）. MethodsAtherosclerosis was induced in 8-week-old male ApoE^-/- mice with high-fat diet. Intragastric administration of SAL was given to one mice group to investigate the effects of SAL on aortic plaque burden， plasma NO level， the migration and angiogenic capabilities of bone marrow-derived EPCs （BM-EPCs）. The proliferation， migration and vasculogenic properties of EPCs isolated from As mice were investigated in vitro. AMPK-sh-RNA or the AMPK inhibitor Compound C was used to investigate the role of AMPK/Akt/eNOS pathway in the regulatory effects of SAL. ResultsCompared with As group， NO level was significantly elevated in SAL group. The sizes of atherosclerotic plaques at the aortic root were reduced with smaller lipid cores in SAL group compared with As group. Moreover， the migration and angiogenesis capacity of EPCs markedly decreased in As mice， while SAL treatment reversed these impairments. Incubation with SAL at concentrations of 20， 40， and 80 μmol/L for 48 hours significantly promoted the proliferation， migration， and angiogenesis of EPCs. AMPK-sh-RNA transfection abrogated the 20 μmol/L SAL improvement in EPC biological activities. Western blot analysis further demonstrated that treatment with Compound C blocked the activation of AMPK/Akt/eNOS signaling pathway induced by SAL. ConclusionSAL upregulates the biological functions of EPCs through activating the AMPK/Akt/eNOS signaling pathway， thereby ameliorating EPC dysfunction during the pathological progression of atherosclerosis.

Objective: To investigate the effect of different pre-stretching on the force degradation of elastomeric chains methods and provide experimental evidence for its application in clinical orthodontic treatment. Methods: One control group (no pre-stretched treatment) and 15 experimental groups (pre-stretched to 25%, 50%, and 100% of the original length in 1-5 cycles) were set up by taking five loop lengths of short elastomeric chain and long elastomeric chain samples, respectively. The elastomeric chains from both the control and experimental groups were mounted on the testing fixture at identical initial lengths. To simulate tooth movement, the fixation device was adjusted to reduce the distance by 0.35 mm weekly. The remaining force of each group was measured at different time points over a 4-week period. Further, the experiment was conducted in a simulated salivary environment with additional thermal cycling. Results: Pre-stretching length and cycles showed a significant interaction effect on the remaining force of elastomeric chains (P<0.001). For short elastomeric chains, the groups pre-stretched to 25% (P=0.001) and 50% (P=0.005) of the original length for 3 cycles showed significantly higher remaining force values than the control group at 24 h. At 7, 14, 21, and 28 d, the groups pre-stretched to 25%, 50%, and 100% of the original length for 3 cycles consistently maintained higher remaining force values than the control group (P<0.001). Within the groups of 3 pre-stretching cycles, no significant differences in remaining force were observed among the 50% group and the 25% (P=0.617) or 100% (P=0.229) groups at 7 d. However, at 14 d, the remaining force of the 50% group was significantly higher than that of both the 25% (P=0.003) and 100% (P=0.027) groups. At 21 d, the 50% group maintained a significantly higher remaining force compared to the 100% group (P=0.020), but the difference compared to the 25% group did not reach statistical significance (P=0.052). By 28 d, the remaining force of the 50% group remained significantly higher than that of both the 25% (P=0.003) and 100% (P=0.003) groups. For long elastomeric chains, the 50% pre-stretch (3 cycles) group showed significantly higher remaining force values than the control group at 8 h (P=0.005) and 24 h (P<0.001). Throughout the experimental period at 7, 14, 21, and 28 d, the remaining force values of the 25% (7, 14 d: P<0.001; 21 d: P=0.003; 28 d: P=0.008), 50% (P<0.001), and 100% (P<0.001) original length pre-stretching for 3 cycles groups were consistently higher than those of the control group. Within the groups of 3 pre-stretching cycles, the remaining force of the 50% original length pre-stretching for 3 cycles group was significantly higher than that of the 25% original length pre-stretching for 3 cycles group at all of the time points from 7 to 28 d (P<0.001). Although no statistically significant differences were observed between the 50% and 100% groups at 14 d (P=0.068), the 50% group exhibited significantly higher remaining force than the 100% group at 7 d (P<0.001), 21 days (P=0.005), and 28 days (P<0.001). Conclusion Appropriate pre-stretching methods can effectively delay the force degradation of the elastomeric chain. Pre-stretching to 50% original length for three cycles can maximize the delay of the force degradation of the elastomeric chain.

Objective: To establish a risk assessment system for infectious disease prevention and control in schools in Shangcheng District, Hangzhou and determine risk levels for each school, and propose corresponding risk management measures, so as to provide a scientific reference for infectious disease prevention and control in primary and secondary schools. Methods: Based on the Failure Mode and Effects Analysis (FMEA) method, potential failure analysis and current situation investigation of infectious disease prevention and control risks were conducted in 110 primary and secondary schools from 2022 to 2024 in Shangcheng District, Hangzhou. Risk levels were classified using K-Means cluster analysis. Results: Through expert panel discussions using FMEA, 6 first level indicators and 28 second level indicators were identified. The top three risk priority numbers were implementation of required prevention and control measures for clustered infectious disease outbreaks in schools in the past three years ( 189.00 ), student morning/afternoon health checks (168.00), and reporting status of clustered infectious disease outbreaks in schools in the past three years (144.00). The comprehensive prevention scores of schools ranged from 61.00 to 98.00 (mean: 87.40 ). There were no statistically significant differences in the average scores(primary school: 88.17±7.39, nine year consistent education: 86.26±7.68, junior high school: 85.55±8.20, and high school: 88.72±4.91) and risk level distribution of schools with different educational stages( F/H=0.95,1.47, P >0.05).K-Means cluster analysis divided the schools into 5 risk levels with cluster centers at 93.25, 85.78, 79.69, 70.29, 61.00 ( F=309.21, P <0.05), with 80% of schools classified as low risk or below. Conclusion The infectious disease prevention and control risk assessment system for primary and secondary schools can be established, and hierarchical management can be conducted according to school risk levels, thereby improving the efficiency and effectiveness of school infectious disease prevention and control, and enhancing the precision and sustainability of prevention efforts.

Interferon regulatory factor 4 (IRF4) is a critical transcription factor that governs the differentiation of cluster of differentiation 4⁺ (CD4⁺) T cells. The pathogenesis and progression of psoriasis are primarily attributed to an immune imbalance stemming from the overproduction of interleukin-17A (IL-17A) by T lymphocytes. However, the role of IRF4 in psoriasis remains unexplored. In this study, we found that IRF4 activity is increased in the cutaneous lesions of patients with psoriasis in response to stimulation by IL-23A and IL-1β. This IRF4 elevation heightens its binding to the E1A binding protein p300 (EP300) promoter, triggering the transcription of downstream retinoic acid receptor-related orphan receptor-γt (RORγt) and increasing the secretion of IL-17A, thereby establishing the IL-1β/IL-23A-IRF4-EP300-RORC-IL-17A inflammatory cascade in psoriasis. The alleviation of imiquimod (IMQ)-induced psoriatic-like symptoms was achieved through the creation of a Irf4 ^-/- gene deletion mouse model and pharmacological inhibition using antisense oligonucleotides targeted for Irf4. This amelioration was accompanied by a decreased number of IL-17A-producing CD4⁺ T cells in the skin. The findings of this study suggest that IRF4 plays a crucial role in the promotion of inflammation and exacerbation of IMQ-induced psoriasiform dermatitis. Consequently, IRF4 targeting could be a promising therapeutic strategy.

OBJECTIVES: To explore the therapeutic mechanism of puerarin for alleviating synovitis in rats with collagen-induced arthritis (CIA). METHODS: In a SD rat model of CIA, we tested the effects of daily gavage of puerarin at low, moderate and high doses (10, 30, and 100 mg/kg, respectively) for 3 weeks, with tripterygium glycosides (GTW, 10 mg/kg) as the positive control, on swelling in the hind limb joints regions evaluated by arthritis index scoring. Mass fraction of the liver of the rats was calculated, and pathologies in joint synovial membrane were observed with HE staining. The expressions of transforming growth factor β‑activated kinase-1 (TAK1), Toll-like receptor 4 (TLR4), and nuclear factor kappa-Bp65 (NF‑κB p65) at the mRNA and protein levels in the synovial tissues were detected using Real-time PCR and Western blotting. RESULTS: Compared with those in the model group, the rats in GTW group and high-dose puerarin group showed significantly reduced mass fraction of the liver. Treatment with GTW and puerarin at the 3 doses all significantly alleviated plantar swelling, lowered arthritis index scores, and improved synovitis in CIA rats (P<0.05), and the effects of puerarin showed an obvious dose dependence. Both GTW and puerarin treatments significantly lowered TAK1, TLR4, and NF‑κB p65 mRNA and protein expressions in the synovium of CIA rats. CONCLUSIONS Puerarin alleviates synovium damages in CIA rats possibly by suppressing the TLR4/NF‑κB signaling pathway via downregulating TAK1 expression.
Animals ; Toll-Like Receptor 4/metabolism* ; Rats, Sprague-Dawley ; Rats ; MAP Kinase Kinase Kinases/metabolism* ; Signal Transduction/drug effects* ; Arthritis, Rheumatoid/drug therapy* ; NF-kappa B/metabolism* ; Isoflavones/therapeutic use* ; Male ; Arthritis, Experimental/drug therapy* ; Transcription Factor RelA/metabolism* ; Synovial Membrane/metabolism*

Stem cells play a crucial role in maintaining tissue regenerative capacity and homeostasis. However, mechanisms associated with stem cell senescence require further investigation. In this study, we conducted a proteomic analysis of human dental pulp stem cells (HDPSCs) obtained from individuals of various ages. Our findings showed that the expression of NUP62 was decreased in aged HDPSCs. We discovered that NUP62 alleviated senescence-associated phenotypes and enhanced differentiation potential both in vitro and in vivo. Conversely, the knocking down of NUP62 expression aggravated the senescence-associated phenotypes and impaired the proliferation and migration capacity of HDPSCs. Through RNA-sequence and decoding the epigenomic landscapes remodeled induced by NUP62 overexpression, we found that NUP62 helps alleviate senescence in HDPSCs by enhancing the nuclear transport of the transcription factor E2F1. This, in turn, stimulates the transcription of the epigenetic enzyme NSD2. Finally, the overexpression of NUP62 influences the H3K36me2 and H3K36me3 modifications of anti-aging genes (HMGA1, HMGA2, and SIRT6). Our results demonstrated that NUP62 regulates the fate of HDPSCs via NSD2-dependent epigenetic reprogramming.
Humans ; Dental Pulp/cytology* ; Nuclear Pore Complex Proteins/genetics* ; Cellular Senescence/genetics* ; Stem Cells/metabolism* ; Epigenesis, Genetic ; Cell Proliferation ; Cell Differentiation ; Histone-Lysine N-Methyltransferase/metabolism* ; Cells, Cultured ; Cellular Reprogramming ; Cell Movement ; Proteomics

Objective:To investigate the application of blended team-based learning (BTBL) in improving the clinical reasoning ability of general practitioners from different sources.Methods:From September to December in 2021, 52 postgraduates of general practice in Tongji University School of Medicine were selected as subjects, and according to their study background and practice experience, they were divided into residency training group, community group, and specialist group. BTBL was adopted for all students in the course of clinical reasoning in general practice, and a 4-station objective structured clinical examination (OSCE) test was conducted before and after the course to evaluate teaching effectiveness from the aspects of medical history collection, physical examination, doctor-patient communication, and medical record writing. SPSS 22.0 was used to perform a statistical analysis of data. Continuous data were expressed as mean±standard deviation, and an analysis of variance or the Kruskal-Wallis test was used for comparison between groups; the paired samples t-test or the Wilcoxon signed rank sum test was used for comparison of normally distributed or non-normally distributed data within each group. Categorical data were expressed as frequency and percentage. Results:There was a significant change in the score of physical examination after the course in the community group (44.88±9.17 vs. 78.06±12.11, t=-12.49, P<0.001), the specialist group (45.85±8.18 vs. 68.65±13.10, t=-6.43, P<0.001), and the residency training group (42.38±15.30 vs. 76.44±16.46, t=-5.98, P<0.001). There was a significant change in the score of doctor-patient communication after the course in the community group (63.00±13.84 vs. 78.69±7.10, t/Z=-4.33, P<0.05) and the residency training group (60.00±15.50 vs. 70.88±6.44, t/Z=-2.12, P<0.05). There was also a significant change in the score of medical record writing after the course in the community group (60.19±17.96 vs. 79.69±14.25, t=-2.96, P<0.05), the specialist group (47.77±12.59 vs. 80.31±12.93, t=-6.26, P<0.05), and the residency training group (37.00±15.50 vs. 75.88±15.88, t=-7.25, P<0.05). The score of medical record writing before the course was 60.19±17.96 in the community group, 47.77±12.59 in the specialist group, and 37.00±15.50 in the residency training group, with a significant difference between the three groups ( χ2=13.04, P=0.001); after the course, the score of medical record writing was 78.69±7.10 in the community group, 63.46±19.40 in the specialist group, and 70.88±6.44 in the residency training group, with a significant difference between the three groups ( χ2=10.13, P=0.006). Conclusions:In the course of clinical reasoning in general practice, BTBL can improve teaching efficiency, save teaching resources, and play the role of promoting strengths and avoiding weaknesses.

The wide application of assisted reproductive technology (ART) has brought the hope of fertility to infertile couples worldwide. However, with the gradual popularization of ART, the long-term health status of ART children has attracted extensive attention from society. Adolescence is a critical developmental period for individuals to transition from childhood to adulthood, with significant physical and psychological changes, so the health status of ART offspring during this period has a profound impact on their overall future health. This article reviews the existing studies, focusing on the effects of ART on multiple aspects of adolescent health, especially growth and development, cardiovascular health, endocrine function, fertility, cancer risk, atopic diseases, mental health, epigenetic changes and other aspects. The possible mechanisms leading to these health problems, including genetic factors, environmental exposures, and the long-term effects of early life interventions, were explored. Future research should continue to explore the long-term effects of ART on the health of offspring and further clarify the mechanisms of these effects, so as to provide a scientific basis for the development of effective prevention and intervention measures.

Objective To analyze and compare the efficacy of DNA barcode,i.e.,Cytochrome b(Cytb)and 12S ribosomal RNA(12S rRNA)gene sequences,in the species identification of wildlife.Methods DNA extraction,quantification,PCR amplification of Cytb and 12S rRNA gene fragments,Sanger sequencing,and sequence alignment analysis were performed on ten wildlife samples.Results Both gene fragments were successfully amplified in six samples,while Cytb alone was successfully amplified in 1 sample,and 12S rRNA alone in 3 samples.Sequence analysis indicated that Cytb enabled species-level identification for 6 samples(Gallinula chloropus,Streptopelia orientalis,Phasianus colchicus,Falco naumanni,Myiopsitta monachus and Lynx lynx)and genus-level identification for 1 sample(Lepus).In contrast,12S rRNA achieved species-level identificaggion for 8 samples(Gallinula chloropus,Lepus sinensis,Phasianus colchicus,Myiopsitta monachus,Muntiacus reevesi,Macaca mulatta and Lynx lynx),representing seven species,and genus-level identification for 1 sample(Falco).However,by combining Cytb and 12S rRNA,all samples could be identified to the species level.Conclusion When applying DNA barcodes to wildlife identification,the Cytb and 12S rRNA gene regions analyzed here can effectively identify common species such as Gallinula chloropus and Streptopelia orientalis,but face difficulties in distinguishing closely related species within the same genus.Therefore,when conducting wildlife species identification,it is recommended to use two or more DNA barcode markers.