Objective To design and synthesize derivatives of oleanolic acid and ursolic acid, and investigate their anti-hepatitis C virus （HCV） activity along with that of common triterpenoid acids. To explore the structure-activity relationship and provide a reference for the research of anti-HCV drugs derived from natural products through obtaining compounds with higher activity. Methods Oleanolic acid and ursolic acid were directly reacted with corresponding amines using PyBOP as a condensing agent in the presence of DIEA. Alternatively, the target compounds were prepared through PCC oxidation followed by the Baeyer-Villiger reaction catalyzed by m-CPBA. In vitro anti-HCV activity was tested using the HCVcc infection model. Molecular docking was performed by Autodock software to investigate the interaction between the active compounds and HCV NS5B. Results Oleanolic acid, glycyrrhetinic acid, ursolic acid, and asiatic acid all exhibited certain anti-HCV effects. Specifically, oleanolic acid derivatives OA2-OA4, OA6, and OA7, as well as ursolic acid derivatives UA1 and UA2, demonstrated superior anti-HCV activity compared to their parent compounds. Preliminary structure-activity relationship analysis revealed that introducing a bulky group to 28-COOH of oleanolic acid and ursolic acid enhanced their activity. Molecular docking results demonstrated that the active compounds could stably bind to HCV NS5B, thereby exhibiting antiviral activity. Conclusion Pentacyclic triterpenoids possessed anti-HCV effects, and their derivatives coud be synthesized to obtain more active compounds. The anti-HCV mechanism of these compounds may be associated with their inhibition of NS5B.

OBJECTIVE: To investigate the effect of human adipose-derived mesenchymal stem cells (hASCs) exosomes on osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) extracted from osteoporotic mice, and to evaluate the effect of hASCs exosomes on preventing bone loss induced by estrogen deficiency. METHODS: hASCs exosomes were extracted by ultracentrifugation. The osteoporotic mice were established by bilateral ovariectomy (OVX). BMSCs were isolated from osteo-porotic mice and cultured for further analysis. In the experimental group, these BMSCs were exposed to an osteogenic induction medium supplemented with hASCs exosomes to evaluate their potential effects on osteogenesis. In contrast, the control group was treated with the same osteogenic induction medium, but without the addition of hASCs exosomes, to serve as a baseline comparison for the study. To comprehensively assess the osteogenic differentiation of BMSCs influenced by hASCs exosomes, alkaline phosphatase (ALP) staining, ALP activity quantitative analysis and quantitative reverse transcription polymerase chain reaction (qPCR) were performed. These evaluations provided critical insights into the role of hASCs exosomes in promoting osteoblast differentiation and bone formation in osteoporotic conditions. The fluorescence labeled hASCs exosomes were injected via the tail vein to observe the biodistribution of exosomes. Two weeks after OVX, the mice were divided into three groups: The experimental group consisted of estrogen-deficient mice receiving hASCs exosome injections; the negative control group consisted of estrogen-deficient mice receiving phosphate-buffered saline (PBS) injections; and the positive control group consisted of mice that underwent Sham surgery and received PBS injections.The injections were administered once every 3 days, for a total of 8 injections. Afterward, the femurs were collected from the mice, and micro-computed tomography (micro-CT) was performed to measure bone mineral density and conduct bone morphometric analysis. RESULTS: hASCs exosomes were successfully extracted using ultracentrifugation. After the induction by hASCs exosomes, ALP staining and ALP activity in the BMSCs extracted from osteoporotic mice were significantly enhanced, the expression of osteogenesis related genes in BMSCs were significantly up-regulated. More trabecular bone and higher bone mineral density were observed in estrogen-deficient mice injected with hASCs exosomes compared with estrogen-deficient mice injected with PBS, and there was no significant decrease in bone mineral density compared with the Sham operation group. CONCLUSION hASCs exosomes promoted the osteogenic differentiation of BMSCs extracted from osteoporotic mice. hASCs exosomes prevented bone loss induced by estrogen deficiency.
Animals ; Mesenchymal Stem Cells/cytology* ; Exosomes ; Estrogens/deficiency* ; Humans ; Osteogenesis ; Cell Differentiation ; Female ; Mice ; Osteoporosis/prevention & control* ; Ovariectomy ; Adipose Tissue/cytology* ; Cells, Cultured

OBJECTIVE: To compare the registration accuracy of cone beam computed tomography (CBCT) images while registering to virtual craniodentofacial patients based on soft tissue and the dentition registration method. METHODS: Virtual dentofacial patients out of 13 selected participants who needed CBCT scanning were established by impression with a registered-block impression (RBI) based on digital dental images, three-dimensional (3D) facial images and maxillofacial CBCT images. CBCT images were processed in the Mimics software program, establishing the craniofacial virtual patients based on CBCT images (CCTs). Registration between virtual patients from RBI and CCT, using the soft tissue in lower half face (STE) and dentition (DTN) as the reference area, respectively, forming two kinds of virtual craniofacial patients based on digital dental images, 3D facial images and skeletal images of CBCT (hiding the soft tissue and dental casts from CBCT). Three-dimensional deviation analysis was performed in the upper half face and lower half face of facial images from CBCT between two kinds of virtual craniodentofacial patients and compared with 3D facial images from RBI and recorded as root mean square error (RMSE). Paired-t test was used to compare the deviations of RMSEs between the upper and lower half of the face and the upper half of the face of facial images from CCT, respectively, between the two kinds of virtual craniodentofacial patients based on STE and DTN methods. RESULTS: Paired-t tests showed that there was no statistically significant difference between the upper and lower half faces of facial images from CCT between STE and DTN (P>0.05), but the deviation of RMSEs of the upper half face of facial images from CCT in STE was smaller than those in DTN [(1.696±0.420) mm vs. (1.752±0.424) mm, P < 0.01]. CONCLUSION The registration accuracy of CBCT registered in virtual craniodentofacial patients using soft tissue as the reference area was higher.
Humans ; Cone-Beam Computed Tomography/methods* ; Imaging, Three-Dimensional/methods* ; Male ; Face/anatomy & histology* ; Female ; Adult ; Image Processing, Computer-Assisted/methods* ; Young Adult ; User-Computer Interface

Evading host immunity killing is a critical step for virus survival. Inhibiting viral immune escape is crucial for the treatment of viral diseases. Serine/threonine kinase 39 (STK39) was reported to play an essential role in ion homeostasis. However, its potential role and mechanism in viral infection remain unknown. In this study, we found that viral infection promoted STK39 expression. Consequently, overexpressed STK39 inhibited the phosphorylation of interferon regulatory factor 3 (IRF3) and the production of type I interferon, which led to viral replication and immune escape. Genetic ablation or pharmacological inhibition of STK39 significantly protected mice from viral infection. Mechanistically, mass spectrometry and immunoprecipitation assays identified that STK39 interacted with PPP2R1A (a scaffold subunit of protein phosphatase 2A (PP2A)) in a kinase activity-dependent manner. This interaction inhibited DDB1 and CUL4 associated factor 1 (DCAF1)-mediated PPP2R1A degradation, maintained the stabilization and phosphatase activity of PP2A, which, in turn, suppressed the phosphorylation of IRF3, decreased the production of type I interferon, and then strengthened viral replication. Thus, our study provides a novel theoretical basis for viral immune escape, and STK39 may be a potential therapeutic target for viral infectious diseases.

Digital technologies have become an integral part of complete denture restoration. With advancement in computer-aided design and computer-aided manufacturing (CAD/CAM), tools such as intraoral scanning, facial scanning, 3D printing, and numerical control machining are reshaping the workflow of complete denture restoration. Unlike conventional methods that rely heavily on clinical experience and manual techniques, digital technologies offer greater precision, predictability, and efficacy. They also streamline the process by reducing the number of patient visits and improving overall comfort. Despite these improvements, the clinical application of digital complete denture restoration still faces challenges that require further standardization. The major issues include appropriate case selection, establishing consistent digital workflows, and evaluating long-term outcomes. To address these challenges and provide clinical guidance for practitioners, this expert consensus outlines the principles, advantages, and limitations of digital complete denture technology. The aim of this review was to offer practical recommendations on indications, clinical procedures and precautions, evaluation metrics, and outcome assessment to support digital restoration of complete denture in clinical practice.
Humans ; Denture, Complete ; Computer-Aided Design ; Denture Design/methods* ; Consensus ; Printing, Three-Dimensional

Objective To construct a curriculum system for higher vocational education in speech-language-hearing rehabilitation based on the World Health Organization rehabilitation competency framework(RCF),to align with international standards. Methods Based on RCF,curriculum theory and principles of vocational education psychology,a curriculum system and its content for higher vocational education in speech-language-hearing rehabilitation were developed.The curric-ulum content and core competencies were analyzed in detail across four levels:basic courses,specialized basic courses,specialized core courses and practical training courses. Results From the perspective of the seven competency domains of RCF,the theoretical and practical significance of con-structing a speech-language-hearing higher vocational education curriculum system based on RCF was systemati-cally elaborated.The study emphasized the importance of establishing competency-oriented higher vocational re-habilitation education for developing students'professional competence,adapting to job requirements,and pro-moting career development.A curriculum content system for speech-language-hearing rehabilitation higher voca-tional education based on RCF was systematically constructed,encompassing basic courses,professional cours-es,and practical training courses.The course names,main content and competency objectives at each level were analyzed in detail. Conclusion It is important to develop the course system of higher vocational education of speech-language-hearing reha-bilitation based on RCF.By comparing with international standards,the curriculum is optimized,focusing on im-proving students'vocational competence and promoting the international development.The curriculum system covers key areas such as core values,beliefs,practice and professionalism,integrates basic,core and practical training courses to achieve a competence-oriented curriculum system for higher vocational speech-language-hear-ing rehabilitation education.

Objective To establish a three-dimensional finite element model of knee joint in patients with mild knee osteoarthritis in the absence of CT and MRI with thin layer thickness,and to analyze the stress distributions of the tibial plateau.Methods A female patient with mild knee osteoarthritis was selected as the study object,and received serial CT scan from pelvis to distal tibia of fibula.The obtained files in DICOM format were imported into Mimics software for extracting the bony structures of knee joint through threshold segmentation,and then optimizing the extracted structures with Edit Masks,Cavity Fill and Region Grow.The generated model of bony structures was imported into 3-matic for smoothing and wrapping,and Geomagic Wrap was used for bony structure processing with mesh doctor inspection,surface defect repair and surface fitting.The cartilages,meniscus and ligaments were established by contour extension,and assembled with bony structures in SolidWorks in the way of origin coincidence.After defining material properties and contact relationships,meshing,and setting constraints and loads in ANSYS software,the stress distributions of the tibial plateau when standing with both legs were analyzed.Results A complete knee joint model including bony structure,cartilage and ligament was successfully established.When standing with two legs,the peak value of contact stress was about 1.21 MPa,and the maximum stress was 0.72 MPa at the medial middle of tibial plateau and lateral tibial plateau.The 62.7%and 37.3%of the total load were borne by the inner and outer compartments.Conclusion The model of the total knee joint is established with CT extraction and software rendering,and the finite element analysis results meet clinical expectations,indicating that the model established by the proposed method is reliable and can be used for follow-up research.

Objective To compare the stability of multi-rod structures with double-headed screws and traditional connectors in posterior three-column osteotomy of the spine using finite element analysis.Methods A finite element model of the T3-L4 thoracolumbar spine was constructed based on postoperative computed tomography(CT)data of patients with severe kyphosis.Based on the patient's standard two-rod model(2R),a double-headed screw multi-rod structure model(4R-DHS)and a traditional connector multi-rod structure model(4R-TC)were constructed.The two models were evaluated under 300 N follower load and 7.5 N-m moment load,and the stability,maximum von Mises stresses on the main rods,and stress distributions of the two multi-rod structures were analyzed.Results There was little difference in the stability between the two multi-rod structures.Compared with 4R-TC,4R-DHS showed a decrease in the maximum von Mises stresses on the main rods during all motions(the stress decreased by 7.2％,8.8％,8.7％,18.5％,and 16.9％during flexion,left lateral bending,right lateral bending,left axial rotation,and right axial rotation,respectively)and more uniform stress distribution,except for a slight increase in the maximum von Mises stresses on the main rods during post-extension.Conclusions The double-headed screw multi-rod structure can reduce the maximum stress on the main rod compared with the traditional connector multi-rod structure,and there is no problem with stress concentration on the main rod near the connector,which can more effectively reduce the risk of internal fixation failure.

Objective:This study aims to investigate the anxiety-like behavior induced by cystitis and its neurobio-logical mechanisms in the anterior cingulate cortex(ACC)by constructing a mouse model of cystitis.Methods:A cys-titis model was established using lipopolysaccharide(LPS).Anxiety-like behavior in mice was explored through the open field test,and single-nucleus RNA sequencing was used to analyze changes in gene expression of neurons in the ACC.Results:HE staining results indicated significant inflammatory responses in the bladders of LPS-treated mice,including extensive infiltration of inflammatory cells,tissue edema,and bladder epithelial shedding.The open field test showed that the time and distance of mice moving in the central area of the open field were significantly reduced,indica-ting obvious anxiety-like behavior.Quality control of single-nucleus sequencing data suggested good data quality.Di-mensionality reduction clustering of the top 2000 highly variable genes revealed seven different cell subpopulations in the ACC.Five major cell types were identified in the ACC,and neurons were further clustered into five different subpopula-tions,identifying two main cell types.Differential gene analysis indicated significant changes in gene expression in neu-rons,with functional enrichment in pathways related to synaptic dysfunction.Conclusion:Cystitis can induce changes in gene expression in specific cell subpopulations in the ACC,suggesting that neurons may play an important role in the anxiety-like behavior induced by cystitis.This provides a new perspective for understanding the relationship between cystitis and anxiety behavior and offers important experimental evidence for future exploration of the brain regions and molecular mechanisms underlying cystitis-induced anxiety behavior.