1.Awareness and attitudes of elderly Southeast Asian adults towards telehealth during the COVID-19 pandemic: a qualitative study.
Ryan Eyn Kidd MAN ; Aricia Xin Yi HO ; Ester Pei Xuan LEE ; Eva Katie Diana FENWICK ; Amudha ARAVINDHAN ; Kam Chun HO ; Gavin Siew Wei TAN ; Daniel Shu Wei TING ; Tien Yin WONG ; Khung Keong YEO ; Su-Yen GOH ; Preeti GUPTA ; Ecosse Luc LAMOUREUX
Singapore medical journal 2025;66(5):256-264
INTRODUCTION:
We aimed to understand the awareness and attitudes of elderly Southeast Asians towards telehealth services during the coronavirus disease 2019 (COVID-19) pandemic in this study.
METHODS:
In this qualitative study, 78 individuals from Singapore (51.3% female, mean age 73.0 ± 7.6 years) were interviewed via telephone between 13 May 2020 and 9 June 2020 during Singapore's first COVID-19 'circuit breaker'. Participants were asked to describe their understanding of telehealth, their experience of and willingness to utilise these services, and the barriers and facilitators underlying their decision. Transcripts were analysed using thematic analysis, guided by the United Theory of Acceptance Use of Technology framework.
RESULTS:
Of the 78 participants, 24 (30.8%) were able to describe the range of telehealth services available and 15 (19.2%) had previously utilised these services. Conversely, 14 (17.9%) participants thought that telehealth comprised solely home medication delivery and 50 (51.3%) participants did not know about telehealth. Despite the advantages offered by telehealth services, participants preferred in-person consultations due to a perceived lack of human interaction and accuracy of diagnoses, poor digital literacy and a lack of access to telehealth-capable devices.
CONCLUSION
Our results showed poor overall awareness of the range of telehealth services available among elderly Asian individuals, with many harbouring erroneous views regarding their use. These data suggest that public health education campaigns are needed to improve awareness of and correct negative perceptions towards telehealth services in elderly Asians.
Humans
;
COVID-19/epidemiology*
;
Female
;
Telemedicine
;
Aged
;
Male
;
Singapore/epidemiology*
;
Qualitative Research
;
Health Knowledge, Attitudes, Practice
;
SARS-CoV-2
;
Aged, 80 and over
;
Middle Aged
;
Pandemics
;
Awareness
;
Asian People
;
Southeast Asian People
2.Adolescent self-harm and suicide attempts: An analysis of emergency department presentations in Singapore.
Darren Kai Siang CHONG ; Vicknesan Jeyan MARIMUTTU ; Pei Shan HOE ; Chu Shan Elaine CHEW ; Angelina Su Yin ANG
Annals of the Academy of Medicine, Singapore 2025;54(2):78-86
INTRODUCTION:
The rising rate of adolescent suicide, and the burden of self-harm and mental health disorders, pose significant threats to Singapore's future health outcomes and human potential. This study sought to examine the risk profile and healthcare utilisation patterns of Singaporean adolescents who presented to the emergency department (ED) for suicidal or self-harm behaviour.
METHOD:
A retrospective review of medical records for patients aged 10 to 19 years who visited Singapore's KK Women's and Children's Hospital ED for suicidal or self-harm attempts from January to December 2021 was conducted.
RESULTS:
A total of 221 patients were identified, with a predominance of female patients (85.5%) over males (14.5%). The mean age was 14.2 ± 1.4 years. Intentional drug overdose (52.0%) was the most commonly used method. Significantly more females presented for intentional paracetamol overdose (46.6% versus [vs] 28.1%, P=0.049), whereas jumping from a height was more common among males (18.8% vs 5.8%, P=0.022). The most frequently observed mental health challenges were stress-related and emotional coping difficulties (50.7%), followed by mood and anxiety symptoms (53.4%). A history of self-harm and suicidal behaviours were the most common psychosocial risk factors. Within the year prior to their ED presentation, 15.4% had accessed healthcare services for mild medical ailments, 19.5% for medically unexplained symptoms, and 17.2% for previous self-harm or suicide attempts.
CONCLUSION
Most cases involved psychosocial and emotional regulation difficulties, some of which displayed sex-specific patterns, rather than complex psychiatric disorders. The identified predictive factors can help inform Singapore's National Mental Health and Well-being Strategy, to guide targeted and transdiagnostic interventions in schools and community settings.
Humans
;
Adolescent
;
Singapore/epidemiology*
;
Female
;
Male
;
Suicide, Attempted/psychology*
;
Emergency Service, Hospital/statistics & numerical data*
;
Self-Injurious Behavior/psychology*
;
Retrospective Studies
;
Child
;
Young Adult
;
Drug Overdose/epidemiology*
;
Risk Factors
;
Acetaminophen/poisoning*
;
Patient Acceptance of Health Care/statistics & numerical data*
;
Sex Factors
3.Singapore consensus statements on the management of obstructive sleep apnoea.
Leong Chai LEOW ; Chuen Peng LEE ; Sridhar VENKATESWARAN ; Michael Teik Chung LIM ; Oon Hoe TEOH ; Ruth CHANG ; Yam Cheng CHEE ; Khai Beng CHONG ; Ai Ping CHUA ; Joshua GOOLEY ; Hong Juan HAN ; Nur Izzianie KAMARUDDIN ; See Meng KHOO ; Lynn Huiting KOH ; Shaun Ray Han LOH ; Kok Weng LYE ; Mark IGNATIUS ; Yingjuan MOK ; Jing Hao NG ; Thun How ONG ; Chu Qin PHUA ; Rui Ya SOH ; Pei Rong SONG ; Adeline TAN ; Alvin TAN ; Terry TAN ; Jenny TANG ; David TAY ; Jade TAY ; Song Tar TOH ; Serene WONG ; Chiang Yin WONG ; Mimi YOW
Annals of the Academy of Medicine, Singapore 2025;54(10):627-643
INTRODUCTION:
Obstructive sleep apnoea (OSA) is common in Singapore, with moderate to severe OSA affecting around 30% of residents. These consensus statements aim to provide scientifically grounded recommendations for the management of OSA, standar-dise the management of OSA in Singapore and promote multidisciplinary collaboration.
METHOD:
An expert panel, which was convened in 2024, identified several areas of OSA management that require guidance. The expert panel reviewed the current literature and developed consensus statements, which were later independently voted on using a 3-point Likert scale (agree, neutral or disagree). Consensus (total ratings of agree and neutral) was set a priori at ≥80% agreement. Any statement not reaching consensus was excluded.
RESULTS:
The final consensus included 49 statements that provide guidance on the screening, diagnosis and management of adults with OSA. Additionally, 23 statements on the screening, diagnosis and management of paediatric OSA achieved consensus. These 72 consensus statements considered not only the latest clinical evidence but also the benefits and harms, resource implications, feasibility, acceptability and equity impact of the recommendations.
CONCLUSION
The statements presented in this paper aim to guide clinicians based on the most updated evidence and collective expert opinion from sleep specialists in Singapore. These recommendations should augment clinical judgement rather than replace it. Management decisions should be individualised, taking into account the patient's clinical characteristics, as well as patient and caregiver concerns and preferences.
Humans
;
Sleep Apnea, Obstructive/diagnosis*
;
Singapore
;
Consensus
;
Adult
4.Clinicopathological significance and prognostic value of serum 25-hydroxyvitamin D3 level in children with IgA vasculitis nephritis.
Pao YU ; Pei ZHANG ; Chun-Lin GAO ; Zi WANG ; Yin ZHANG ; Zheng GE ; Bi ZHOU
Chinese Journal of Contemporary Pediatrics 2025;27(1):55-61
OBJECTIVES:
To study the significance of serum 25-hydroxyvitamin D3 [25-(OH)D3] level in the clinicopathological characteristics and prognosis of children with immunoglobulin A vasculitis nephritis (IgAVN).
METHODS:
A retrospective analysis was conducted on the clinical data of children with IgAVN who underwent renal biopsy at Suzhou Hospital Affiliated to Anhui Medical University and Jinling Hospital of the Medical School of Nanjing University from June 2015 to June 2020. Based on serum 25-(OH)D3 level, the patients were divided into a normal group and a lower group. The clinicopathological characteristics and follow-up data of the two groups were collected and compared.
RESULTS:
A total of 359 children with IgAVN were included. Compared to the normal group (62 cases), the lower group (297 cases) exhibited higher incidences of hematochezia and gross hematuria, higher levels of serum creatinine, blood urea nitrogen, urinary retinol protein, urinary N-acetyl-β-D-glucosaminidase, and quantitative urinary protein, and a longer duration from renal biopsy to urinary protein becoming negative, as well as lower estimated glomerular filtration rate and albumin level (P<0.05). Renal pathology in the lower group showed a higher occurrence of tubular interstitial injury, crescent formation, segmental sclerosis in glomeruli, and inflammatory cell infiltration in the renal interstitium compared to the normal group (P<0.05). Survival analysis indicated that the cumulative renal survival rate was lower in the lower group (P<0.05). Multivariate Cox regression analysis revealed that low serum 25-(OH)D3 level is an independent risk factor for poor prognosis in children with IgAVN.
CONCLUSIONS
Children with IgAVN and low serum 25-(OH)D3 level have relatively severe clinicopathological manifestations. Low serum 25-(OH)D3 level is an independent risk factor for poor prognosis in children with IgAVN.
Humans
;
Male
;
Female
;
Child
;
Prognosis
;
Retrospective Studies
;
Calcifediol/blood*
;
Child, Preschool
;
Adolescent
;
Glomerulonephritis, IGA/mortality*
;
Vasculitis/pathology*
;
IgA Vasculitis/mortality*
5.Preparation and Evaluation of Clinical-Grade Human Umbilical Cord-Derived Mesenchymal Stem Cells with High Expression of Hematopoietic Supporting Factors.
Jie TANG ; Pei-Lin LI ; Xiao-Yu ZHANG ; Xiao-Tong LI ; Fu-Hao YU ; Jia-Yi TIAN ; Run-Xiang XU ; Bo-Feng YIN ; Li DING ; Heng ZHU
Journal of Experimental Hematology 2025;33(3):892-898
OBJECTIVE:
To prepare clinical-grade human umbilical cord-derived mesenchymal stem cells (hUC-MSC) with high expression of hematopoietic supporting factors and evaluate their stem cell characteristics.
METHODS:
Fetal umbilical cord tissues were collected from healthy postpartum women during full-term cesarean section. Wharton's jelly was mechanically separated and hUC-MSCs were obtained by explant culture method and enzyme digestion method in an animal serum-free culture system with addition of human platelet lysate. The phenotypic characteristics of hUC-MSCs obtained by two methods were detected by flow cytometry. The differences in proliferation ability between the two groups of hUC-MSCs were identified through CCK-8 assay and colony forming unit-fibroblast (CFU-F) assay. The differences in multilineage differentiation potential between the two groups of hUC-MSCs were identified through induction of adipogenic, osteogenic, and chondrogenic differentiation. The mRNA expression levels of hematopoietic supporting factors such as SCF, IL-3, CXCL12, VCAM1 and ANGPT1 in the two groups of hUC-MSCs were identified by real-time fluorescence quantiative PCR(RT-qPCR).
RESULTS:
The results of flow cytometry showed that hUC-MSCs obtained by the two methods both expressed high levels of CD73, CD90 and CD105, while lowly expressed CD31, CD45 and HLA-DR. The results of CCK-8 and CFU-F assay showed that the proliferation ability of hUC-MSCs obtained by explant culture method was better than those obtained by enzyme digestion method. The results of the triple lineage differentiation experiment showed that there was no significant difference in multilineage differentiation potential between the two grous of hUC-MSCs. The results of RT-qPCR showed that the mRNA expression levels of hematopoietic supporting factors SCF, IL-3, CXCL12, VCAM1 and ANGPT1 in hUC-MSCs obtained by explant cultrue method were higher than those obtained by enzyme digestion method.
CONCLUSION
Clinical-grade hUC-MSCs with high expression levels of hematopoietic supporting factors were successfully cultured in an animal serum-free culture system.
Humans
;
Mesenchymal Stem Cells/metabolism*
;
Umbilical Cord/cytology*
;
Cell Differentiation
;
Female
;
Cell Proliferation
;
Cells, Cultured
;
Chemokine CXCL12/metabolism*
;
Angiopoietin-1/metabolism*
;
Vascular Cell Adhesion Molecule-1/metabolism*
;
Stem Cell Factor/metabolism*
;
Flow Cytometry
;
Pregnancy
6.Research Progress of Vagal Nerve Regulation Mechanism in Acupuncture Treatment of Atrial Fibrillation.
Lu-Lu CAO ; Hui-Rong LIU ; Ya-Jie JI ; Yin-Tao ZHANG ; Bing-Quan WANG ; Xiao-Hong XUE ; Pei WANG ; Zhi-Hui LUO ; Huan-Gan WU
Chinese journal of integrative medicine 2025;31(3):281-288
Atrial fibrillation (AF) is the most common arrhythmia in clinical practice. It has a high prevalence and poor prognosis. The application of antiarrhythmic drugs and even surgery cannot completely treat the disease, and there are many sequelae. AF can be classified into the category of "palpitation" in Chinese medicine according to its symptoms. Acupuncture has a significant effect on AF. The authors find that an important mechanism of acupuncture in AF treatment is to regulate the cardiac vagus nerve. Therefore, this article intends to review the distribution and function of vagus nerve in the heart, the application and the regulatroy effect for the treatment of AF.
Atrial Fibrillation/physiopathology*
;
Humans
;
Acupuncture Therapy
;
Vagus Nerve/physiology*
;
Animals
7.Outcomes of identifying enlarged vestibular aqueduct (Mondini malformation) related gene mutation in Mongolian people
Jargalkhuu E ; Tserendulam B ; Maralgoo J ; Zaya M ; Enkhtuya B ; Ulzii B ; Ynjinlhkam E ; Chuluun-Erdene Ts ; Chen-Chi Wu ; Cheng-Yu Tsai ; Yin-Hung Lin ; Yi-Hsin Lin ; Yen-Hui Chan ; Chuan-Jen Hsu ; Wei-Chung Hsu ; Pei-Lung Chen
Mongolian Journal of Health Sciences 2025;87(3):8-15
Background:
Hearing loss (HL) is one of the most common sensory disorders,
affecting over 5-8% of the world's population. Approximately half of HL cases are
attributed to genetic factors. In hereditary deafness, about 75-80% is inherited
through autosomal recessive inheritance, and common pathogenic genes include
GJB2 and SLC26A4. Pathogenic variants in the SLC26A4gene are the leading
cause of hereditary hearing loss in humans, second only to the GJB2 gene. Variants in the SLC26A4gene cause hearing loss, which can be non-syndromic autosomal recessive deafness (DFNB4, OMIM #600791) associated with enlarged
vestibular aqueduct (EVA) or Pendred syndrome (Pendred, OMIM #605646).
DFNB4 is characterized by sensorineural hearing loss combined with EVA or less
common cochlear malformation defect. Pendred syndrome is characterized by bilateral sensorineural hearing loss with EVA and an iodine defect that can lead to
thyroid goiter. Currently, it is known that EVA is associated with variants in the
SLC26A4 gene and is a penetrant feature of SLC26A4-related HL. Predominant
mutations in these genes differ significantly across populations. For instance, predominant SLC26A4 mutations differ among populations, including p.T416P and
c.1001G>A in Caucasians, p.H723R in Japanese and Koreans, and c.919-2A>G
in Han Taiwanese and Han Chinese. On the other hand, there has been no study
of hearing loss related to SLC26A4 gene variants among Mongolians, which is the
basis of our research.
Aim:
We aimed to identify the characteristics of the SLC26A4 gene variants in
Mongolian people with Enlarged vestibular aqueduct and Mondini malformation.
Materials and Methods:
In 2022-2024, We included 13 people with hearing loss
and enlarged vestibular aqueduct, incomplete cochlea (1.5 turns of the cochlea
with cystic apex- incomplete partition type II- Mondini malformation) were examined by CT scan of the temporal bone in our study. WES (Whole exome sequencing) analysis was performed in the Genetics genetic-laboratory of the National
Taiwan University Hospital.
Results:
Genetic analysis revealed 26 confirmed pathogenic variants of bi-allelic
SLC26A4 gene of 8 different types in 13 cases, and c.919-2A>G variant was dominant with 46% (12/26) in allele frequency, and c.2027T>A (p.L676Q) variant 19%
(5/26), c.1318A>T(p.K440X) variant 11% (3/26), c.1229C>T (p.T410M) variant 8%
(2/26) ) , c.716T>A (p.V239D), c.281C>T (p.T94I), c.1546dupC, and c.1975G>C
(p.V659L) variants were each 4% (1/26)- revealed. Two male children, 11 years
old (SLC26A4: c.919-2A>G) and 7 years old (SLC26A4: c.919-2A>G:, SLC26A4:
c.2027T>A (p.L676Q))had history of born normal hearing and progressive hearing
loss.
Conclusions
1. 26 variants of bi-allelic SLC26A4 gene mutation were detected
in Mongolian people with EVA and Mondini malformation, and c.919-2A>G was
the most dominant allele variant, and rare variants such as c.1546dupC, c.716T>A
(p.V239D) were detected.
2. Our study shows that whole-exome sequencing (WES) can identify gene
mutations that are not detected by polymerase chain reaction (PCR) or NGS analysis.
8.Causal relationship between five autoimmune diseases and intervertebral disc degeneration:a bidirectional Mendelian analysis.
Bao-Fei ZHANG ; Xun-Lu YIN ; Ze-Ling HUANG ; Shuai PEI ; Yu-Wei LI
China Journal of Orthopaedics and Traumatology 2025;38(8):856-866
OBJECTIVE:
To explore the potential causal relationship between intervertebral disc degeneration and certain autoimmune diseases.
METHODS:
Genome-wide association study (GWAS) data of 5 autoimmune diseases were obtained from large-scale GWAS databases. Data on internal vertebral disc degeneration (IVDD) were derived from the FinnGen consortium, which included 294, 770 controls and 41, 669 cases. A two-sample bidirectional Mendelian randomization (MR) analysis was performed to investigate the potential causal relationship between the 5 autoimmune diseases and IVDD. Multiple analytical methods were adopted, including MR methods such as inverse variance weighting(IVW), MR-Egger, weighted median, weighted mode, and simple mode. Cochran's Q test, leave-one-out analysis, and MR-Egger intercept test were conducted to assess heterogeneity, robustness, and pleiotropy. For the robustness of the results, MR-PRESSO was used to detect outliers, and MR analysis was re-conducted after removing the outliers.
RESULTS:
The MR analysis results showed that there might be a bidirectional causal relationship between ankylosing spondylitis(AS) and IVDD:AS on IVDD, OR=1.038, 95%CI (1.024, 1.053), P=0.000;and IVDD on AS, OR=2.117, 95%CI(1.065, 4.207), P=0.032. There might be a positive correlation between IVDD and rheumatoid arthritis(RA) as well as systemic lupus erythematosus(SLE):IVDD on RA, OR=1.184, 95%CI(1.071, 1.309), P=0.001;and IVDD on SLE, OR=1.678, 95%CI(1.187, 2.372), P=0.003. There was no significant correlation between ulcerative colitis(UC), autoimmune thyroiditis(ATD) and IVDD. After removing outliers by MR-PRESSO and re-conducting MR analysis, the results did not change qualitatively. Sensitivity analysis indicated that the results were robust to potential sources of bias.
CONCLUSION
AS and IVDD may be risk factors for each other, and IVDD may be a potential risk factor for RA and SLE. These findings provide a basis for guiding the prevention and combined diagnosis and treatment of IVDD, AS, RA, and SLE, while the specific underlying mechanisms still require further experimental basic research.
Humans
;
Intervertebral Disc Degeneration/etiology*
;
Mendelian Randomization Analysis
;
Autoimmune Diseases/complications*
;
Genome-Wide Association Study
;
Spondylitis, Ankylosing/genetics*
;
Arthritis, Rheumatoid/genetics*
9.Mechanism of inhibiting miR-34a-5p expression and promoting bone growth in mouse brain tissue by Semen Ziziphi Spinosae extract.
Yuan-Yuan PEI ; Yan XIE ; Na YIN ; Wen-Long MA ; Wei-Peng XING ; Gui-Zhi WANG ; Qing-Feng WANG
China Journal of Orthopaedics and Traumatology 2025;38(10):1061-1070
OBJECTIVE:
To explore the mechanism by which the extract of Semen Ziziphi Spinosae extract promotes bone growth in mice by modulation of the expression of miR-34a-5p in brain tissue.
METHODS:
Mice were assigned to four experimental groups:a normal control group, a drug administration group (receiving 0.320 mg·g-1 body weight of Semen Ziziphi Spinosae extract via intragastric administration), a positive control group (receiving 0.013 mg·g-1 body weight of jujube seed saponin via intragastric administration), and a combination group administration with Semen Ziziphi Spinosae extract plus a 5-hydroxytryptamine 2A receptor (5-HT2AR) agonist (intragastric administration of Semen Ziziphi Spinosae extract combined with intracerebroventricular injection of 8 μg P-MPPF per mice for the final three days of the experiment). Following a 20-day administration period, the effects of the interventions on bone growth, serum growth hormone (GH) levels, and 5-HT2AR expression in brain tissue were evaluated. MicroRNAs (miRNAs) that were differentially expressed in the brain tissues of mice exhibiting bone growth induced by Semen Ziziphi Spinosae extract, as compared to those in normal mice, were identified using a gene chip approach. The interaction between miR-34a-5p and 5-HT2AR was subsequently validated through quantitative reverse transcription polymerase chainreaction (RT-qPCR) and dual-luciferase reporter gene assays. Subsequently, by utilizing the miR-34a-5p inhibitor group and mimics group, along with the normal control group, the drug administration group, the positive control group, and the drug administration combined with miR-34a-5p inhibitor group, the variations in 5-HT2AR expression in mouse brain tissue across all groups were examined, and the binding activity of 5-hydroxytryptamine (5-HT) to the 5-hydroxytryptamine 1A receptor (5-HT1AR) in mice was assessed.
RESULTS:
The body lengths of the normal control group and the drug administration group were(8.9±0.3) and(10.4±0.4) cm;femur lengths were (8.5±0.3) and (9.1±0.5) mm;tibia lengths were (10.7±0.3) and (11.2±0.4) mm, respectively. The contents of GH levels were (58.6±8.2) and (72.9±6.1) ng·ml-1;and the contents of 5-HT2AR were (32.0±5.0) and (21.9± 5.5) ng·ml-1, respectively. Compared with the normal control group, the drug administration group promoted the growth of body length, femur, and tibia in mice, and increased GH secretion, showing statistically significant differences (P<0.05). Additionally, it significantly reduced the content of 5-HT2AR in brain tissue, with statistical significance (P<0.01). The gene chip analysis identified a total of 16 differentially expressed miRNAs, of which 13 were up-regulated and 3 were down-regulated. Bioinformatics analysis predicted that the up-regulated miR-34a-5p could regulate the expression of 5-HT2AR, a prediction that was confirmed through a dual-luciferase reporter gene assay, demonstrating a direct regulatory interaction between the two. Furthermore, in vivo experiments in mice revealed that overexpression and silencing of miR-34a-5p resulted in corresponding changes in the expression levels of 5-HT2AR in brain tissues/cells, as well as in the binding activity between 5-HT and 5-HT1AR.
CONCLUSION
The Semen Ziziphi Spinosae extract promotes animal bone growth by enhancing miR-34a-5p expression in brain tissue, downregulating the expression level of 5-HT2AR, improving the binding activity between 5-HT and 5-HT1AR, and extending slow-wave sleep duration, thereby stimulating GH secretion.
Animals
;
MicroRNAs/metabolism*
;
Mice
;
Male
;
Brain/metabolism*
;
Ziziphus/chemistry*
;
Bone Development/drug effects*
;
Drugs, Chinese Herbal/pharmacology*
;
Plant Extracts/pharmacology*
10.Active ingredient combination of Longma formula promotes chondrocyte proliferation via regulating Wnt/β-catenin signaling pathway
Zi-han PEI ; Yin-sheng CAO ; Qi-yao ZHANG ; Kun JIANG ; Gong-jin ZHOU ; Ke-fang DONG ; Ping WU
Chinese Pharmacological Bulletin 2025;41(9):1776-1782
Aim To investigate the promotive effects and mechanisms of the combined use of brucine(Bru)and lumbrokinase(LK),active ingredient derived from Longma formula,in promoting chondrocyte proliferation via the Wnt/β-catenin signaling pathway.Methods The extracted primary rat chondrocytes were divided in-to the following groups:Control group,Bru,LK alone group,and Bro+LK combination group.The optimal drug concentration and intervention time were deter-mined using CCK-8 assay,followed by cell proliferation validation through EdU and phalloidin staining.The expression levels of collagen Ⅱ,aggrecan and SRY-re-lated high-mobility group box gene 9(SOX9)in chon-drocytes following intervention with the combination of Bru and LK were detected by Western blotting.Addi-tionally,the regulatory effects of these proteins on the Wnt/β-catenin signaling pathway were also investiga-ted.Results The optimal combination concentration of Longma formula active ingredients(Bru 0.025 mg·L-1+LK 5 mg·L-1)significantly enhanced chondro-cyte viability compared to control,Bru,or LK alone at 48 h.This combination increased the S-phase ratio,promoted the aggregation of intracellular actin fila-ments,and upregulated the expression of collagen Ⅱ and aggrecan.Furthermore,it activated the Wnt/β-catenin pathway,leading to increased SOX9 expres-sion.Conclusions The optimal combination of Bru and LK(Bru 0.025 mg·L-1+LK 5 mg·L-1)de-rived from Longma formula significantly maintains chondrocyte phenotype and promotes cellular prolifera-tion through the activation of the Wnt/β-catenin signa-ling pathway,which subsequently upregulates the downstream target SOX9.

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