1.Efficacy Connotation and Mechanisms of Shudi Qiangjin Pills Against Steroid-induced Osteonecrosis of Femoral Head Based on "Disease-Syndrome-Formula" Association Network
Zhijian CHEN ; Suya ZHANG ; Longlong DING ; Guixin ZHANG ; Bo LIU ; Baohong MI ; Yanqiong ZHANG ; Na LIN ; Weiheng CHEN ; Chunzhu GONG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):88-99
ObjectiveTo elucidate the efficacy connotation of Shudi Qiangjin pills (SQP) against liver and kidney deficiency in steroid-induced osteonecrosis of femoral head (SONFH) from the perspective of the "disease-syndrome-formula" association and to clarify the underlying mechanisms based on in vivo and in vitro experiment validation. MethodsThe chemical components and the corresponding putative targets of SQP were collected from the Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine (TCMIP) v2.0, the Encyclopedia of Traditional Chinese Medicine (ETCM) v2.0, and HERB databases. The SONFH-related genes were identified based on the differential expression profiles of peripheral blood of patients with SONFH compared to the healthy volunteers, and the disease phenotype-related targets were collected from the TCMIP v2.0 database. Then, the interaction network of "SONFH-related genes and candidate targets of SQP" was constructed based on "gene-gene interaction information", and the major network targets were screened by calculating the topological characteristic values of the network followed by the functional mining according to the Kyoto Encyclopedia of Genes and Genomes (KEGG) database and the SoFDA database. After that, the SONFH rat model was prepared by lipopolysaccharide combined with methylprednisolone injection, and 2.5, 5, 7.5 g·kg-1 SQP (once per day, equivalent to 1, 2, and 3 times the clinical equivalent dose, respectively) or 7.3×10-3 g·kg-1 of alendronate sodium (ALS, once per week, equivalent to the clinical equivalent dose) was given for 8 weeks. The effect characteristics of SQP and ALS in the treatment of SONFH were evaluated by micro-computed tomography scanning, hematoxylin and eosin staining, alkaline phosphatase (ALP) staining, immunohistochemical staining, enzyme-linked immunosorbent assay, and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)staining, and a comparative efficacy analysis was conducted with ALS. In addition, SONFH cell models were prepared by dexamethasone stimulation of osteoblasts, and the intervention was carried out with the medicated serum of SQP at the aforementioned three doses. Cell counting kit-8, ALP staining, ALP activity assay, alizarin red staining, and flow cytometry were employed to investigate the regulatory effect of SQP on osteoblasts. The expression levels of osteogenesis-related proteins and key factors of the target signaling axis were detected by quantitative real-time polymerase chain reaction and Western blot. ResultsThe network analysis results demonstrated that the candidate targets of SQP primarily exerted their therapeutic effects through key signaling pathways, including phosphoinositide 3-kinase(PI3K)/protein kinase B(Akt), lipid metabolism and atherosclerosis, prolactin, chemokines, and neurotrophic factors pathways. These pathways were significantly involved in critical biological processes such as muscle and bone metabolism and the regulation of the "neuro-endocrine-immune" network, thereby addressing both modern medical symptoms (e.g., delayed skeletal maturation and recurrent fractures) and traditional Chinese medicine (TCM) symptoms (e.g., fatigue, aversion to cold, cold limbs, and pain in the limbs and joints in patients with SONFH characterized by liver and kidney deficiency syndrome. Among these pathways, the PI3K/Akt signaling pathway exhibited the highest degree of enrichment. The in vivo experimental results demonstrated that starting from the 4th week after modeling, the modeling group exhibited a significant reduction in body weight compared to the control group (P<0.05). After six weeks of treatment, all dosage groups of SQP showed significantly higher body weights compared to the model group (P<0.01). Compared with the normal group, the model group exhibited significant decreases in bone mineral density (BMD), bone volume fraction (BV/TV), trabecular number (Tb.N), osteocalcin (OCN), alkaline phosphatase (ALP) levels in femoral head tissue, and serum bone-specific alkaline phosphatase (BALP) (P<0.01), along with significant increases in trabecular separation (Tb.Sp), empty lacunae rate in tissue, and apoptosis rate (P<0.01). In comparison to the model group, the SQP intervention groups showed significant improvements in BMD, BV/TV and Tb.N (P<0.01), significant reductions in Tb.Sp, empty lacunae rate and apoptosis rate (P<0.05), and significant increases in protein levels of OCN and ALP as well as BALP content (P<0.05). The in vitro experimental results revealed that all dosage groups of SQP medicated serum showed no toxic effects on osteoblast. Compared with the normal group, the model group displayed significant suppression of osteoblast proliferation activity, ALP activity, and calcified nodule formation rate (P<0.01), significant decreases in mRNA transcription levels of OCN and Runt-related transcription factor 2 (RUNX2) (P<0.01), significant reductions in protein content of osteopontin (OPN), typeⅠ collagen (ColⅠ)A1, B-cell lymphoma-2 (Bcl-2), PI3K, and phosphorylated (p)-Akt (P<0.01), and a significant increase in apoptosis rate (P<0.01). Compared with the model group, the SQP medicated serum intervention groups exhibited significant increases in proliferation activity, ALP activity, calcified nodule formation rate, mRNA transcription levels of OCN and RUNX2, and protein content of OPN, ColⅠA1, Bcl-2, PI3K, and p-Akt (P<0.05), along with a significant decrease in apoptosis rate (P<0.01). ConclusionSQP can effectively reduce the disease severity of SONFH with liver and kidney deficiency syndrome and improve bone microstructure, with the therapeutic effects exhibiting a dose-dependent manner. The mechanism may be related to its regulation of key processes such as muscle and bone metabolism and the correction of imbalances in the "neuro-endocrine-immune" network, thereby promoting osteoblast differentiation and inhibiting osteoblast apoptosis. The PI3K/Akt signaling axis is likely one of the key pathways through which this formula exerts its effects.
2.Current status of eating behaviors and its predictive role in overweight and obese of adolescents
Chinese Journal of School Health 2025;46(1):53-57
Objective:
To explore the current status and influencing factors of eating behaviors in adolescents, so as to provide a theoretical foundation for health promotion education among adolescents.
Methods:
Based on the database from Survey of Chinese Family Health Index (2021), by a random number table method, 1 065 teenagers were selected from the provincial capitals of 22 provinces and 5 autonomous regions in China, as well as 4 municipalities directly under the central government. A general characteristic questionnaire, Patient Health Questionnaire-9 (PHQ-9), Short Form of the Family Health Scale (FHS-SF), 10-item Short Version of the Big Five Personality(BFP-10), Content-based Media Exposure Scale (CM-E) and Sakata Eating Behavior Scale Short Form(EBS-SF) were used to collect information. Multivariate stepwise linear regression analysis was employed to identify and analyze related factors of eating behaviors among adolescents. Receiver operating characteristic was used to validate the predictive ability of the EBS-SF score for overweight and obesity among adolescents.
Results:
The average scores of BFI-10,C-ME, FHS-SF, PHQ-9 and EBS-SF were (33.08±4.64)(19.20±4.55)(38.48±6.65)(6.09±5.63)(16.75±4.36), respectively. Multivariate linear regression showed that family type (other types), agreeableness, conscientiousness, family health and depression were the main related factors of EBS-SF scores among adolescents( B =2.61,-0.42,0.20,-0.11,0.23, P <0.05).The analysis of receiver operating characteristic curve showed that the EBS-SF scores had a good ability in predicting obesity among male adolescents ( AUC= 0.73, P <0.01).
Conclusions
Family type, big five personality, family health,depression are the related factors of eating behaviors among adolescents. EBS-SF scores are predictive of obesity in adolescents, which would provide a new perspective for promoting healthy eating habits among adolescents.
3.Visualization analysis of artificial intelligence in bone trauma research based on Citespace
Haoran SONG ; Yuqiang ZHANG ; Na GU ; Xiaodong ZHI ; Wei WANG
Chinese Journal of Tissue Engineering Research 2025;29(3):493-502
BACKGROUND:The development of artificial intelligence in the medical field is rapidly advancing,with increasing research on its applications in the field of bone trauma.Through bibliometric analysis,this paper analyzed the research hotspots of artificial intelligence in the field of bone trauma in recent years,and predicted the future research trend. OBJECTIVE:To summarize the development history,research status,hot spots,and future development trends of artificial intelligence technology in the field of bone trauma to provide new insights for future research. METHODS:This study selected relevant literature from the Web of Science core database,covering the period from the inception to August 2023,and retrieved 420 articles related to the application of artificial intelligence,machine learning,and deep learning in the field of bone trauma.After manual screening,202 articles related to this article were exported,and Citespace software was used for visual analysis of cooperation of countries,institutions,cited journals,citation analysis,keyword co-occurrence,and other aspects. RESULTS AND CONCLUSION:(1)The overall number of publications from the 202 selected articles showed an upward trend,indicating significant research potential for future studies.The country with the highest centrality and the highest publication volume was the United States.The University of California(USA)was the most prolific research institution.(2)The top five most commonly used keywords in bone trauma research using artificial intelligence were deep learning,artificial intelligence,bone density,machine learning,and diagnosis.The keyword with the highest centrality was bone density,and the keyword with the highest frequency was deep learning.(3)The top 10 most cited reference papers provided comprehensive insights into the feasibility of applying artificial intelligence techniques to the diagnosis of bone trauma from various perspectives.Among them,eight papers focused on bone and joint injuries and deep convolutional neural networks.One paper discussed the use of deep learning in detecting osteoporosis in CT scans to prevent fragility fractures,while another paper explored the correlation between the application of artificial intelligence in identifying changes in skin texture and the recognition of bone characteristics.(4)In the future,the research hotspots of artificial intelligence will mainly focus on the specific study of fractures caused by bone and joint trauma and osteoporosis.The research trend mainly focuses on improving the performance of artificial intelligence algorithms,using new artificial intelligence technologies to accurately classify and quickly and efficiently diagnose bone injuries,especially for the diagnosis of complex and hidden fractures.By establishing finite element analysis models,more standardized evaluations of bone injuries can be achieved.
4.SR9009 combined with indolepropionic acid alleviates inflammation in C2C12 myoblasts through the nuclear factor-kappa B signaling pathway
Huihui JI ; Xu JIANG ; Zhimin ZHANG ; Yunhong XING ; Liangliang WANG ; Na LI ; Yuting SONG ; Xuguang LUO ; Huilin CUI ; Ximei CAO
Chinese Journal of Tissue Engineering Research 2025;29(6):1220-1229
BACKGROUND:Rev-erbα is involved in the regulation of inflammation,but pharmacological activation of Rev-erbα increases the risk for cardiovascular diseases.To reduce the relevant risk,an exploration on SR9009,a Rev-erbα agonist,combined with other drugs to relieve inflammation in skeletal myoblasts was conducted,laying the theoretical foundation for the treatment of inflammation-associated skeletal muscle atrophy. OBJECTIVE:To investigate the relationship of SR9009,indolepropionic acid and nuclear factor-κB signaling pathways in lipopolysaccharide-induced C2C12 myoblasts. METHODS:(1)C2C12 myoblasts were induced to differentiate in the presence of lipopolysaccharide(1 μg/mL).RNA-seq and KEGG pathway analysis were used to study signaling pathways.(2)C2C12 myoblast viability was assessed using the cell counting kit-8 assay to determine optimal concentrations of indolepropionic acid.Subsequently,cells were categorized into control group,lipopolysaccharide(1 μg/mL)group,SR9009(10 μmol/L)+lipopolysaccharide group,indolepropionic acid(80μmol/L)+lipopolysaccharide group,and SR9009+indolepropionic acid+lipopolysaccharide group.ELISA was employed to measure protein expression levels of interleukin-6 in the cultured supernatant.Real-time quantitative PCR were employed to measure mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.Western blot assay were employed to measure protein expression levels of NF-κB p65 and p-NF-κB p65.(3)After Rev-erbα was knocked down by siRNA,knockdown efficiency was assessed by RT-qPCR.And mRNA levels of interleukin-6 and tumor necrosis factor α were also measured. RESULTS AND CONCLUSION:Compared with the blank control group,lipopolysaccharide time-dependently inhibited myofibroblast fusion to form myotubes,the mRNA expression levels of interleukin-6 and tumor necrosis factor α were elevated,and the level of interleukin-6 in the cell supernatant was significantly increased.The results of KEGG pathway showed that the nuclear factor-κB signaling pathway was activated by lipopolysaccharide.Indolepropionic acid exhibited significant suppression of C2C12 myoblasts viability when its concentration exceeded 80 μmol/L.Indolepropionic acid and SR9009 inhibited the activation of NF-κB signaling pathway,thereby played an anti-inflammatory role,and suppressed the mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.Compared with the lipopolysaccharide group,the ratio of p-NF-κB p65/NF-κB p65 protein expression were downregulated.SR9009 combined with indolepropionic acid notably reduced lipopolysaccharide-induced inflammation,further downregulated the mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.The ratio of p-NF-κB p65/NF-κB p65 protein expression was significantly lower than that in the SR9009+lipopolysaccharide group or indolepropionic acid+lipopolysaccharide group.Rev-erbα increases time-dependently with lipopolysaccharide induction.The knockdown efficiency of Rev-erbα by siRNA reached over 58%,and lipopolysaccharide was added after Rev-erbα was successfully knocked down.Compared with the lipopolysaccharide group,the mRNA expression levels of interleukin-6 and tumor necrosis factor α were significantly up-regulated.These results conclude that Rev-erbα may act as a promising pharmacological target to reduce inflammation.SR9009 targeted activation of Rev-erbα combined with indolepropionic acid significantly inhibits the nuclear factor-κB signaling pathway and attenuates the inflammatory response of C2C12 myofibroblasts.Moreover,the combined anti-inflammatory effect is superior to that of the intervention alone.
5.Design, synthesis and antifungal and antitumor activity research of novel Hsp90 inhibitors
Qiao SHI ; Guiyan HAN ; Junteng ZHANG ; Na LIU
Journal of Pharmaceutical Practice and Service 2025;43(3):124-135
Objective To design and synthesize novel Hsp90 inhibitors with dual functions of synergistically enhancing the antifungal activity of fluconazole (FLC) against drug-resistant fungi and anti-tumor activity based on the Hsp90 inhibitor Ganetespib. Methods The previous research found that Ganetespib had a good synergistic anti-resistant fungal activity with FLC, with a fractional inhibitory concentration index (FICI) of 0.023 to 0.039. In this study, structural modifications were made to Ganetespib by replacing its indole ring with a phenyl ring containing different substituents to design and synthesize a series of new compounds. The in vitro synergistic anti-resistant fungal activity against C. albicans 0304103 in combination with FLC, anti-tumor activity (against HEL, HL60 and A549 cells), and Hsp90α inhibition activity were determined to explore their structure-activity relationship and mechanism of action. Results The chemical structures of 19 new compounds were confirmed by 1H NMR, 13C NMR and HRMS. Most of the compounds exhibited strong Hsp90α inhibitory activity, good synergistic activity against drug-resistant fungi in combination with FLC and anti-tumor activity. The substitution of electron-donating groups on the benzene ring was beneficial to enhancing the synergistic activity against drug-resistant fungi in combination with FLC. Among them, compounds F3 and F5 showed excellent synergistic activity against drug-resistant fungi in combination with FLC (FICI were both 0.047) and anti-tumor activity (IC50 were 0.025 to 0.15 μmol/L and 0.021 to 0.23 μmol/L respectively), and could down-regulate the expression levels of drug resistance genes and efflux pump genes in fungi, inhibit the formation of fungal biofilms, and arrest the cell cycle of HEL cells at G0/G1 phase. Conclusion The novel Hsp90 inhibitors such as F3 and F5 could both effectively exert the dual activities of synergizing with FLC to combat drug-resistant fungi and fight against tumors, which provided a new idea for the development of new drugs with dual functions of synergizing with FLC to combat drug-resistant fungi and fight against tumors.
6.Interventional Effect and Mechanisms of Renqing Mangjue on MNNG-induced Malignant Transformation of Gastric Mucosal Epithelial Cells
Peiping CHEN ; Fengyu HUANG ; Xinzhuo ZHANG ; Xiangying KONG ; Ziqing XIAO ; Yanxi LI ; Xiaohui SU ; Na LIN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(8):69-77
ObjectiveThis study aimed to investigate the intervention effect of Renqing Mangjue on the malignant transformation of gastric mucosal epithelial cells induced by N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) and to explore its molecular mechanism in preventing precancerous lesions of gastric cancer based on the cyclic guanosine monophosphate (cGMP)/protein kinase G (PKG)/mitogen-activated protein kinase (MEK)/extracellular signal-regulated kinase (ERK) signaling pathway. MethodsHuman gastric mucosal epithelial cells (GES-1) were initially induced by MNNG to establish a precancerous cell model (MC cells). The effective concentration of MNNG for inducing malignant transformation in GES-1 cells was screened using the cell proliferation activity decection (CCK-8) assay, and the effective concentration of Renqing Mangjue for inhibiting the proliferation of transformed GES-1 cells was also determined. GES-1 cells were divided into a blank control group, a model group, and treatment groups with Renqing Mangjue at concentrations of 1, 3, 10, and 30 mg·L-1. Furthermore, the effects of Renqing Mangjue on the migratory ability and epithelial-mesenchymal transition (EMT) characteristics of GES-1 malignant transformed cells were evaluated using Transwell migration assays, wound healing assays, and real-time quantitative reverse transcription polymerase chain reaction (Real-time PCR). Additionally, candidate chemical components and target sites of Renqing Mangjue were obtained from the TCMIP v2.0 database, and disease targets at various stages of gastric cancer precursors were sourced from the Gene Expression Omnibus (GEO) database. Pathway enrichment analysis was performed using the Metascape database to predict the potential mechanisms of action of Renqing Mangjue. Finally, the protective mechanism of Renqing Mangjue against gastric cancer precursors was validated through Western blot analysis. ResultsAt a concentration of 20 μmol·L-1, MNNG exhibited an inhibition rate of approximately 50% on GES-1 cells (P<0.01), and at this concentration, the GES-1 cells displayed biological characteristics indicative of malignant transformation. In contrast, Renqing Mangjue had no significant effect on the proliferation of normal GES-1 cells, but significantly inhibited the proliferation of MC cells (P<0.01) and markedly reduced their migratory capacity (P<0.01). Moreover, it also increased the mRNA expression level of E-cadherin during the EMT process (P<0.05), while inhibiting the expression of both N-cadherin and the transcription factor Snail mRNA (P<0.05, P<0.01). Network predictions suggested that Renqing Mangjue may prevent gastric cancer precursors through modulating the cGMP/PKG and MAPK/ERK signaling pathways. Furthermore, Western blot results indicated that Renqing Mangjue upregulated the expression of PKG and NPRB (B-type natriuretic peptide receptor) proteins in the cGMP/PKG pathway (P<0.01), while downregulating the expression of the downstream proteins MEK and ERK (P<0.05, P<0.01). ConclusionIn summary, Renqing Mangjue can prevent gastric cancer precursors by inhibiting the proliferation and migration of malignant transformed GES-1 cells, thereby delaying the EMT process. The underlying mechanisms may be related to the activation of the cGMP/PKG pathway and the inhibition of the MEK/ERK signaling pathway.
7.Risk Identification Model of Coronary Artery Stenosis Constructed Based on Random Forest
Yongfeng LV ; Yujing WANG ; Leyi ZHANG ; Yixin LI ; Na YUAN ; Jing TIAN
Journal of Sun Yat-sen University(Medical Sciences) 2025;46(1):138-146
ObjectiveTo establish a risk recognition model for coronary artery stenosis by using a machine learning method and to identify the key causative factors. MethodsPatients aged ≥18 years,diagnosed with coronary heart disease through coronary angiography from January 2013 to May 2020 in two prominent hospitals in Shanxi Province, were continuously enrolled. Logistic regression,back propagation neural network (BPNN), and random forest(RF)algorithms were used to construct models for detecting the causative factors of coronary artery stenosis. Sensitivity (TPR), specificity (TNR), accuracy (ACC), positive predictive value (PV+), negative predictive value (PV-), area under subject operating characteristic curve (AUC), and calibration curve were used to compare the discrimination and calibration performance of the models. The best model was then employed to predict the main risk variables associated with coronary stenosis. ResultsThe RF model exhibited superior comprehensive performance compared to logistic regression and BPNN models. The TPR values for logistic regression,BPNN,and RF models were 75.76%, 74.30%, and 93.70%, while ACC values were 74.05%, 72.30%, and 79.49%, respectively. The AUC values were:logistic regression 0.739 9; BPNN 0.723 1; RF 0.752 2. Manifestations such as chest pains,abnormal ST segments on ECG,ventricular premature beats with hypertension, atrial fibrillation, regional wall motion abnormalities(RWMA) by color echocardiography, aortic regurgitation(AR), pulmonary insufficiency (PI), family history of cardiovascular diseases,and body mass index(BMI)were identified as top ten important variables affecting coronary stenosis according to the RF model. ConclusionsRandom forest model shows the best comprehensive performance in identification and accurate assessment of coronary artery stenosis. The prediction of risk factors affecting coronary artery stenosis can provide a scientific basis for clinical intervention and help to formulate further diagnosis and treatment strategies so as to delay the disease progression.
8.Palpitations, Shortness of Breath, Weakness in Limbs, Edema, and Dyspnea: A Rare Inflammatory Myopathy with Positive Aniti-mitochondrial Antibodies and Cardiac Involvement
Chunsu LIANG ; Xuchang ZHANG ; Ning ZHANG ; Lin KANG ; Xiaohong LIU ; Jiaqi YU ; Yingxian LIU ; Lin QIAO ; Yanli YANG ; Xiaoyi ZHAO ; Ruijie ZHAO ; Na NIU ; Xuelian YAN
Medical Journal of Peking Union Medical College Hospital 2025;16(1):248-255
This article presents a case study of a patient who visited the Geriatric Department of Peking Union Medical College Hospital due to "palpitations, shortness of breath for more than 2 years, limb weakness for 6 months, edema, and nocturnal dyspnea for 2 months". The patient exhibited decreased muscle strength in the limbs and involvement of swallowing and respiratory muscles, alongside complications of heart failure and various arrhythmias which were predominantly atrial. Laboratory tests revealed the presence of multiple autoantibodies and notably anti-mitochondrial antibodies. Following a comprehensive multidisciplinary evaluation, the patient was diagnosed with anti-mitochondrial antibody-associated inflammatory myopathy. Treatment involved a combination of glucocorticoids and immunosuppressants, along with resistance exercises for muscle strength and rehabilitation training for lung function, resulting in significant improvement of clinical symptoms. The case underscores the importance of collaborative multidisciplinary approaches in diagnosing and treating rare diseases in elderly patients, where careful consideration of clinical manifestations and subtle abnormal clinical data can lead to effective interventions.
9.Effect of Shenqi Yiliu Formula (参芪抑瘤方) Drug-Containing Serum on the Cycle of Gastric Cancer Cell AGS and Wnt/β-Catenin Pathway
Na WEI ; Chongyuan GUO ; Min BAI ; Yaorong AN ; Sichao ZHANG ; Liyang WU ; Yongqiang DUAN
Journal of Traditional Chinese Medicine 2025;66(4):399-406
ObjectiveTo investigate the potential mechanism of Shenqi Yiliu Formula (参芪抑瘤方) in treating precancerous lesions of gastric cancer (PLGC) by the Wnt/β-catenin signaling pathway. MethodsThe CCK-8 assay was used to determine the optimal intervention time for Shenqi Yiliu Formula drug-containing serum and the concentration of the Wnt/β-catenin pathway inhibitor XAV939 depends on the survival rate of AGS gastric cancer cell line. AGS cells were divided into the gastric cancer cell group (15% blank serum), inhibitor group (selected concentration of XAV939), high-dose Shenqi Yiliu Formula group (12% Shenqi Yiliu Formula drug-containing serum + 3% blank serum), medium-dose Shenqi Yiliu Formula group (6% Shenqi Yiliu Formula drug-containing serum + 9% blank serum), and low-dose Shenqi Yiliu Formula group (3% Shenqi Yiliu Formula drug-containing serum + 12% blank serum). Each group was tested in triplicate. After culturing for 24 and 48 hours, cell migration and invasion were assessed by scratch assays; after a selected intervention period (48 hours), cell cycle distribution was analyzed using flow cytometry, Ki67 protein levels were detected by immunofluorescence, the protein levels of Wnt, β-catenin, GSK-3β, and intranuclear T-cell specific factor(TCF) were measured by the protein immunoblotting assay, and the mRNA expressions of these above factors were determined by quantitative real-time PCR. ResultsThe optimal intervention time for Shenqi Yiliu Formula drug-containing serum was determined to be 48 hours, and the effective concentration of XAV939 was 20 μmol/L. Compared with the gastric cancer cell group, Shenqi Yiliu Formula at all doses reduced the cell migration rate at 24 and 48 hours (P<0.05), except for the low-dose group at 24 hours. Compared to the low-dose group at corresponding time points, high- and medium-dose Shenqi Yiliu Formula groups showed significantly reduced migration rates, particularly the high-dose group at 48 hours (P<0.05). Compared with the gastric cancer cell group, the high-dose Shenqi Yiliu Formula and inhibitor groups exhibited reduced protein and mRNA levels of Wnt, β-catenin, and TCF, along with reduced Ki67 protein levels and a decreased proportion of cells in the S and G2 phases of the cell cycle, but GSK-3β protein levels, GSK-3β mRNA expression, and the proportion of cells in the G1 phase increased (P<0.05). Compared to the inhibitor group, the high-dose Shenqi Yiliu Formula group showed a decreased proportion of G1-phase cells and an increased proportion of G2-phase cells (P<0.05), although differences in Wnt and β-catenin protein levels and mRNA expressions were not statistically significant (P>0.05). ConclusionShenqi Yiliu Formula drug-containing serum inhibits the migration and invasion of gastric cancer AGS cells and block the cell cycle at G1 phase, and its underlying mechanism may be related to the regulation of the Wnt/β-catenin signaling pathway.
10.Guidelines for Establishing Animal Models of Rheumatoid Arthritis with Cold-dampness Obstruction Syndrome and Dampness-heat Obstruction Syndrome
Na LIN ; Yanqiong ZHANG ; Changhong XIAO ; Shenghao TU ; Jianning SUN ; Shijun XU ; Representation Preparation GROUP
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):47-54
Rheumatoid arthritis belongs to arthralgia syndrome in the theory of traditional Chinese medicine, and cold-dampness obstruction syndrome and dampness-heat obstruction syndrome are core syndromes and main syndrome differentiation types of this disease. Fine therapeutic effects have been obtained in the long-term clinical practice of many famous traditional Chinese medicine practitioners following the syndrome differentiation and treatment based on the guiding principles of cold and heat. To adapt to the clinical diagnosis practice of combining disease differentiation and syndrome differentiation, and to better carry out basic research on integrated Chinese and Western medicine and preclinical study on new traditional Chinese medicines, Guidelines for Establishing Animal Models of Rheumatoid Arthritis with Cold-Dampness Obstruction Syndrome and Dampness-Heat Obstruction Syndrome (hereinafter referred to as the Guidelines) were compiled by our research group, in cooperation with the renowned experts in research fields including traditional Chinese medicine, clinical medicine, zoology and evidence-based medicine, which provide a meaningful reference for scientific research, teaching and clinical applications. The compilation process of the Guidelines was guided by the theory of disease and syndrome integration and the principles of "evidence takes the main place, consensus plays an auxiliary role, and experience serves as the reference". Based on the comprehensive evaluation of pathogenesis homology, behavioral phenotypic consistency, and drug treatment predictability compared between animal models and human diseases, by the nominal group method, "recommendations" were formed for recommendations supported by evidence, and "consensus recommendations" were formed for recommendations not supported by evidence. Guidelines were formed involving content such as animal types, arthritis modeling methods, external stimulation conditions, and modeling assessment indicators during the establishment of the animal models of rheumatoid arthritis with cold-dampness obstruction syndrome and dampness-heat obstruction syndrome. The Guidelines are applicable for the disease and syndrome research on rheumatoid arthritis, investigation of therapeutic mechanisms, and development of new traditional Chinese medicine. The Guidelines also provide a reference for the establishment of guidelines on other types of diseases and syndromes combined with animal models to further promote the modernization of traditional Chinese medicine research and its integration with international academic development.


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