1.Association between takeout fast foods and sugar sweetened beverage consumption with co-occurrence of anxiety and depressive symptoms among first year junior high school students in Yunnan Province
HU Dongyue, ZHANG Zhengwu, XU Zenglei, TAO Lei, ZENG Anna, GUAN Liao, CHANG Litao,〖JZ〗 HUANG Xin, CHEN Weiwei, LI Jiangli, XU Honglü ;
Chinese Journal of School Health 2026;47(1):23-26
Objective:
To explore the association between takeout fast foods and sugar sweetened beverage consumption with co-occurrence of anxiety and depressive symptoms among first year junior high school students in Yunnan Province, so as to provide theoretical basis for the prevention of anxiety and depressive symptoms co-occurrence among adolescents.
Methods:
A random cluster sampling involving 8 500 first year junior high school students in 11 counties in Yunnan Province was conducted by a questionnaire survey from October to December 2022. The Depression Anxiety Stress Scale-21 (DASS-21) was applied to assess anxiety and depressive symptoms in first year junior high school students. Chi-square test was used to compare the anxiety-depression co-occurrence symptoms of first year junior high school students with different demographic characteristics. The association between takeout fast foods and sugar sweetened beverage consumption with co-occurrence of anxiety and depressive symptoms of adolescents was analyzed by binary Logistic regression models.
Results:
The detection rate of co-occurrence of anxiety and depression symptoms among first year junior high school students in Yunnan Province was 26.92%. After controlling for demographic variables and other confounders, takeout fast foods and sugar sweetened beverage consumption( OR=1.50, 95%CI =1.27-1.77) was associated with anxiety-depression co-occurrence symptoms among first year junior high school students in Yunnan Province ( P <0.01). Stratified analysis showed that both Han ( OR=1.37, 95%CI =1.07-1.77) and ethnic minorities ( OR=1.60, 95%CI =1.29-2.00) exhibited statistically significant associations between takeout fast foods and sugar sweetened beverage consumption with co-occurrence of anxiety and depressive symptoms(both P <0.05).
Conclusions
Takeout fast foods and sugar sweetened beverage consumption increases the risk of co-occurrence of anxiety and depressive symptoms among first year junior high school students in Yunnan Province. It is recommended to strengthen guidance on the consumption of such products among junior high school students to prevent co-occurrence of anxiety and depressive symptoms.
2.Berberine ameliorates coronary artery endothelial cell injury in Kawasaki disease through complement and coagulation cascades.
Jin-Wen LIAO ; Xin GUO ; Bo LIANG ; Xu-Xia LI ; Ming-Guo XU
Chinese Journal of Contemporary Pediatrics 2025;27(1):101-108
OBJECTIVES:
To explore the role of berberine (BBR) in ameliorating coronary endothelial cell injury in Kawasaki disease (KD) by regulating the complement and coagulation cascade.
METHODS:
Human coronary artery endothelial cells (HCAEC) were divided into a healthy control group, a KD group, and a BBR treatment group (n=3 for each group). The healthy control group and KD group were supplemented with 15% serum from healthy children and KD patients, respectively, while the BBR treatment group received 15% serum from KD patients followed by the addition of 20 mmol/L BBR. Differential protein expression was analyzed and identified using isobaric tags for relative and absolute quantitation technology and liquid chromatography-tandem mass spectrometry, followed by GO functional enrichment analysis and KEGG signaling pathway enrichment analysis of the differential proteins. Western blot was used to detect differential protein expression.
RESULTS:
A total of 518 differential proteins were identified between the KD group and the healthy control group (300 upregulated proteins and 218 downregulated proteins). A total of 422 differential proteins were identified between the BBR treatment group and the KD group (221 upregulated proteins and 201 downregulated proteins). Bioinformatics analysis showed that compared to the healthy control group, the differential proteins in the KD group were enriched in the complement and coagulation cascade and ribosome biogenesis in eukaryotes. Compared to the KD group, the differential proteins in the BBR treatment group were also enriched in the complement and coagulation cascade and ribosome biogenesis in eukaryotes. Western blot results indicated that compared to the healthy control group, the expression of complement C1q subcomponent subunit C (C1QC), kininogen-1 (KNG1), complement C1s subcomponent (C1S), and C4b-binding protein alpha chain (C4BPA) was increased in the KD group (P<0.05). Compared to the KD group, the expression of KNG1, C1S, C1QC, and C4BPA was decreased in the BBR treatment group (P<0.05).
CONCLUSIONS
The complement and coagulation cascade may be involved in the regulation of BBR treatment for coronary injury in KD, and C1QC, KNG1, C1S, and C4BPA may serve as biomarkers for this treatment.
Mucocutaneous Lymph Node Syndrome/blood*
;
Humans
;
Endothelial Cells/pathology*
;
Complement System Proteins/physiology*
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Coronary Vessels/drug effects*
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Male
;
Blood Coagulation/drug effects*
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Berberine/therapeutic use*
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Female
;
Child, Preschool
;
Infant
3.Mechanism of Reactive Oxygen/Nitrogen Species in Liver Ischemia-Reperfusion Injury and Preventive Effect of Chinese Medicine.
Lei GAO ; Yun-Jia LI ; Jia-Min ZHAO ; Yu-Xin LIAO ; Meng-Chen QIN ; Jun-Jie LI ; Hao SHI ; Nai-Kei WONG ; Zhi-Ping LYU ; Jian-Gang SHEN
Chinese journal of integrative medicine 2025;31(5):462-473
Liver ischemia-reperfusion injury (LIRI) is a pathological process involving multiple injury factors and cell types, with different stages. Currently, protective drugs targeting a single condition are limited in efficacy, and interventions on immune cells will also be accompanied by a series of side effects. In the current bottleneck research stage, the multi-target and obvious clinical efficacy of Chinese medicine (CM) is expected to become a breakthrough point in the research and development of new drugs. In this review, we summarize the roles of reactive oxygen species (ROS) and reactive nitrogen species (RNS) in various stages of hepatic ischemia-reperfusion and on various types of cells. Combined with the current research progress in reducing ROS/RNS with CM, new therapies and mechanisms for the treatment of hepatic ischemia-reperfusion are discussed.
Reperfusion Injury/drug therapy*
;
Reactive Oxygen Species/metabolism*
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Reactive Nitrogen Species/metabolism*
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Humans
;
Liver/drug effects*
;
Animals
;
Medicine, Chinese Traditional
;
Drugs, Chinese Herbal/pharmacology*
4.Mechanism of Sangqi Qingxuan Liquid in Alleviating Vascular Endothelial Injury in Hypertension Focuses on β-Catenin.
Wei-Quan REN ; Xin ZENG ; Jiang-Quan LIAO ; Li HUANG ; Lin LI
Chinese journal of integrative medicine 2025;31(8):726-734
OBJECTIVE:
To explore the main components and potential mechanisms of Sangqi Qingxuan Liquid in the treatment of arterial vascular endothelial cells (AVECs) injury in hypertension through network pharmacology.
METHODS:
Traditional Chinese Medicine Systems Pharmacology and Analysis Platform (TCMSP) and Traditional Chinese Medicine Integrated Database (TCMID) were used to screen the active components of Sangqi Qingxuan Liquid (SQQX), which met the oral utilization rate and drug similarity criteria. An active component-target network was constructed using Cytoscape 3.6 software. A protein-protein interaction (PPI) network of targets associated with SQQX treatment for hypertension was constructed using the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) database. The Metascape database was used to perform enrichment analysis of gene ontology biological functions and MSigDB pathway enrichment analysis of proteins in the PPI network. Further analysis of the main components of SQQX was performed using UPLC-MS. Based on the results of network pharmacology, the mechanism of SQQX to improve the injury of AVECs in hypertension was verified through lentiviral transfection by Wnt/ β -catenin signaling pathway. AVECs induced by angiotensin II (Ang II ) was used to establish a model of endothelial function injury in hypertension. Cell viability, intracellular nitric oxide content, malonaldehyde content, and superoxide dismutase activity were measured to determine the optimal induction conditions. The optimal intervention conditions for SQQX were determined based on cell viability, cellular DNA activity, and the gradient method. The cells were further divided into blank, model, overexpression lentivirus negative control, overexpression lentivirus, overexpression lentivirus + SQQX intervention (2.47 mg/mL, 12 h), inhibition lentivirus negative control, inhibition lentivirus, and inhibition lentivirus + SQQX intervention (2.47 mg/mL, 12 h) groups. Finally, quantitative real-time PCR and Western blotting were performed to analyze the molecular mechanisms of SQQX in the Wnt/ β -catenin signaling pathway.
RESULTS:
The main SQQX components were betaine, buddleoside, and chlorogenic acid, in descending order. Network pharmacology analysis screened 12 pathways associated with the hypertensive vascular endothelium. The results showed that 1 µ mol/L for 12 h was the optimal condition for Ang II to induce AVECs injury, and 2.47 mg/mL SQQX intervention for 12 h was the optimal condition for treating AVECs injury. In the experimental validation based on the interaction network of the Wnt/ β -catenin signaling pathway, SQQX significantly decreased the expressions of β -catenin, Smad2, peroxisome proliferator-activated receptors (PPARs), endothelial nitric oxide synthase (eNOS), and endothelin-1 (ET-1) caused by the β -catenin overexpression lentivirus (P<0.05 or P<0.01). The function of vascular endothelial cells can be improved by the β -catenin inhibition lentivirus, and no obvious changes were observed after further intervention with SQQX.
CONCLUSION
SQQX may protect against AVECs injury by regulating the Wnt/β -catenin signaling pathway.
Drugs, Chinese Herbal/therapeutic use*
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beta Catenin/metabolism*
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Hypertension/metabolism*
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Endothelial Cells/metabolism*
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Protein Interaction Maps/drug effects*
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Humans
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Wnt Signaling Pathway/drug effects*
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Network Pharmacology
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Endothelium, Vascular/injuries*
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Cell Survival/drug effects*
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Angiotensin II/pharmacology*
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Nitric Oxide/metabolism*
5.Protective effect of catalpol on liver injury in acute hepatitis B rats by regulating TLR4/NF-κB signaling pathway
Xuexia LIAO ; Xin PENG ; Tingting HUANG ; Guidan XIE
Chinese Journal of Immunology 2025;41(3):545-550
Objective:To investigate the protective effect of catalpol on the liver of rats with acute hepatitis caused by hepatitis B virus(HBV)infection and its possible mechanism.Methods:SD rats were randomly divided into Sham group,Model group,acyclo-vir group(ACY group),low-dose catalpol group(L-CAT group),medium-dose catalpol group(M-CAT group)and high-dose catalpol group(H-CAT group).The levels of serum inflammatory cytokines were detected by ELISA.Flow cytometry was used to detect the per-centage of peripheral blood Th17 cells.Hematoxylin-eosin staining and TUNEL assay were used to detect liver tissue injury and apopto-sis.Western blot analysis was used to detect the expression of related proteins in liver tissue.Results:Compared with the Sham group,the levels of serum HBsAg,HBeAg,ALT,AST,Th17 cell ratio,IL-17 and IL-23,and the expression of TLR4 and p-p65 protein in liver tissues were significantly increased in the Model group,while the levels of serum IL-13 and IL-4 were significantly decreased in the Model group,the differences were statistically significant(P<0.05).Compared with the Model group,the levels of serum HBsAg,HBeAg,ALT,AST,Th17 cell ratio,IL-17 and IL-23,as well as the expressions of TLR4 and p-p65 protein in liver tissues of rats in the ACY,M-CAT and H-CAT groups were significantly decreased,while the levels of serum IL-13 and IL-4 were significantly in-creased,the differences were statistically significant(P<0.05).Compared with the L-CAT and M-CAT groups,the levels of serum HB-sAg,HBeAg,ALT,AST,Th17 cell ratio,IL-17 and IL-23,the expression of liver TLR4 and p-p65 protein were significantly de-creased,while the levels of serum IL-13 and IL-4 were significantly increased in the H-CAT group,the differences were statistically significant(P<0.05).Conclusion:Catalpol improves HBV infection-induced liver injury in rats by reducing Th17 cell proportion and pro-inflammatory cytokine levels,and increasing anti-inflammatory cytokine levels,possibly by inhibiting TLR4/NF-κB p65 signaling pathway activation.
6.Glycosylated ceramide-related metabolic enzymes and atherosclerosis
Xin TAN ; Qiang WEI ; Yuan-hong LIAO ; Jing-kun LU
Chinese Pharmacological Bulletin 2025;41(6):1005-1010
Atherosclerosis is an arterial lesion involving abnor-mal lipid metabolism and an inflammatory response,and is the initiating factor of many cardiovascular and cerebrovascular dis-eases.Glycosylated ceramides are derivatives of ceramide mole-cules with a glycosylated group attached,which not only affect the structural integrity of cell membranes,but also regulate apop-tosis,inflammation and lipid metabolism,and disorders of glyco-sylated ceramide metabolism are closely related to the occurrence and progression of atherosclerosis.This review focuses on the specific functions of the glycosylated ceramide-related metabolic enzymes glucose ceramide synthetase,glucosylceramide syn-thetase,lactose ceramide synthetase,and galactosidase in athero-sclerosis,as well as their important effects on the development of atherosclerosis.Targeted therapeutic strategies for these meta-bolic enzymes,related drug development and significant potential in atherosclerosis prevention and treatment are also reviewed.
7.Occurrence and influencing factors of multiple organ dysfunction syndrome in elderly patients with subarachnoid hemorrhage
Xiuqin YANG ; Lingli ZHANG ; Zhenshen LONG ; Li ZHANG ; Xin LIAO ; Zhongxia LIU
Chinese Journal of Geriatric Heart Brain and Vessel Diseases 2025;27(7):941-945
Objective To explore the occurrence and influencing factors of MODS in elderly pa-tients with SAH.Methods A retrospective analysis was conducted on 187 elderly SAH patients admitted in the Second Affiliated Hospital of Guizhou Medical University from January 2021 to June 2024.According to the presence or absence of MODS,they were divided into MODS group(81 cases)and non-MODS group(106 cases).Their general clinical data were compared between the two groups.Multivariate logistic regression analysis was used to identify the risk factors for MODS in elderly SHA patients.ROC curve analysis was applied to evaluate the predictive value of the major risk factors for MODS in the patients.Results The MODS group had significantly ad-vanced age,larger proportion of modified Fisher grades Ⅲ—Ⅳ,higher APACHE Ⅱ score,and in-creased FPG level than the non-MODS group(P<0.05,P<0.01).Multivariate logistic regression analysis showed that APACHE Ⅱ score(OR=1.108,95%CI:1.070-1.148,P=0.000)and mo-dified Fisher grade(OR=1.143,95%CI:1.095-1.194,P=0.000)were independent risk factors for MODS in elderly SAH patients.ROC curve analysis suggested that the AUC value of APACHE Ⅱ score,modified Fisher grade and their combination in predicting the occurrence of MODS in elderly SAH patients was 0.809,0.737 and 0.880,respectively,with a sensitivity of 59.35%,88.93%and 84.02%,and a specificity of 88.73%,58.52%and 81.15%,respectively.The combined detection showed better predictive performance than the indicator alone(P<0.01).Con-clusion The incidence of MODS is quite high in elderly patients with SAH.APACHE Ⅱ score and modified Fisher grade are the main risk factors affecting the occurrence of MODS.
8.Optimization of 90Y PET/CT imaging based on the block-sequential regularized expectation maximization reconstruction algorithm
Tiantian ZHANG ; Ziwei LIANG ; Zhongbin HANG ; Yan ZHANG ; Deqing LIU ; Yuhang SHAN ; Yong LIAO ; Xin HUANG ; Bin LIANG ; Lin ZHANG ; Xiaobin FENG ; Zuoxiang HE
Chinese Journal of Nuclear Medicine and Molecular Imaging 2025;45(6):335-340
Objective:To optimize the image quality of PET/CT following 90Y-selective internal radiation therapy ( 90Y-SIRT) using block-sequential regularized expectation maximization (BSREM) reconstruction algorithm, and to evaluate its impact of different β values on image quality and quantitative analysis. Methods:A retrospective study was conducted on 8 male patients with hepatic tumors (age: 62(52, 71) years) treated at Beijing Tsinghua Changgung Hospital, School of Clinical Medicine, Tsinghua Medicine Tsinghua University, between June 2024 and January 2025. All patients were treated with 90Y resin microspheres (2.6(0.9, 3.6)GBq) and underwent post-treatment 90Y PET/CT liver imaging. Imaging data were reconstructed using BSREM with different noise penalty weighting factors ( β values: 0, 300, 1000, 1500, 2500, 3500, 4000, 6000, 8000, 10000). Visual assessment was independently performed by two nuclear medicine physicians, using a 4-point scale (1=worst, 4=best). The mean score was considered as the final score. The consistency of the 2 reviewers was calculated and analyzed by Kappa test. Visual scores of different β value groups were compared by Friedman test. The β value demonstrating highest mean score and optimal consistency was selected as the optimal. Quantitative analysis was performed using MIM software to calculate the maximum absorbed dose ( Dmax) and the mean absorbed dose ( Dmean) for tumor, normal liver, and whole liver regions, and the CV was used to evaluate the impact of β values. Results:The visual assessment consistency of reviewers in 3 β value groups (0, 3500, 6000) were the highest (7/8) (all kappa=0.88, all P<0.05). Visual scores of the 10 β value groups were significantly different ( χ2=28.74, P<0.001), and the visual scores of 2 β value groups (3500, 4000) were the highest, both of which were 4.0(4.0, 4.0). Overall, visual assessment identified β=3500 as the optimal. Quantitative analysis revealed that, (1) Dmax in all regions (tumor, normal liver, whole liver) decreased with the increasing β values, stabilizing when β>1000 ( CV 56%-67%); (2) Dmean remained stable across different β values ( CV 0.04%-5.00%). Conclusions:In BSREM reconstruction, β=3500 is the optimal parameter for improving 90Y-PET image quality. β values significantly affect Dmax (stabilizing at β > 1000), but have no significant impact on Dmean, suggesting that reconstruction parameters primarily influence dose distribution morphology rather than average dose assessments.
9.Guideline for diagnosis and treatment of infection after internal fixation of closed lower limb fractures in adults (version 2025)
Bobin MI ; Faqi CAO ; Weixian HU ; Wu ZHOU ; Chenchen YAN ; Hui LI ; Yun SUN ; Yuan XIONG ; Jinmi ZHAO ; Qikai HUA ; Xinbao WU ; Xieyuan JIANG ; Dianying ZHANG ; Zhongguo FU ; Dankai WU ; Guangyao LIU ; Guodong LIU ; Tengbo YU ; Jinhai TAN ; Xi CHEN ; Fengfei LIN ; Zhangyuan LIN ; Dongfa LIAO ; Aiguo WANG ; Shiwu DONG ; Gaoxing LUO ; Zhao XIE ; Dong SUN ; Dehao FU ; Yunfeng CHEN ; Changqing ZHANG ; Kun LIU ; Deye SONG ; Yongjun RUI ; Fei WU ; Ximing LIU ; Junwen WANG ; Meng ZHAO ; Biao CHE ; Bing HU ; Chengjian HE ; Guanglin WANG ; Xiao CHEN ; Guandong DAI ; Shiyuan FANG ; Wenchao SONG ; Ming CHEN ; Guanghua GUO ; Yongqing XU ; Lei YANG ; Wenqian ZHANG ; Kun ZHANG ; Xin TANG ; Hua CHEN ; Weiguo XU ; Shuquan GUO ; Yong LIU ; Xiaodong GUO ; Zhewei YE ; Liming XIONG ; Tian XIA ; Hongbin WU ; Qisheng ZHOU ; Mengfei LIU ; Yiqiang HU ; Yanjiu HAN ; Hang XUE ; Kangkang ZHA ; Wei CHEN ; Zhiyong HOU ; Bin YU ; Jiacan SU ; Peifu TANG ; Baoguo JIANG ; Guohui LIU
Chinese Journal of Trauma 2025;41(5):421-432
Postoperative infection of internal fixation of closed fractures the lower limbs in adults represents a devastating complication, characterized by diagnostic challenges, prolonged treatment duration and high disability rates. Current management of these infections faces multiple challenges, such as difficulties in early accurate diagnosis, and various controversies about the treatment plan, leading to poor overall diagnosis and treatment results. To address these issues, based on evidence-based medicine and principles with emphasis on scientific rigor, clinical applicability and innovation, the Trauma Branch of the Chinese Medical Association, Orthopedic Branch of the Chinese Medical Doctor Association, Orthopedics Branch of the Chinese Medical Association, and Trauma Orthopedics and Polytrauma Group of the Resuscitation and Emergency Committee of the Chinese Medical Doctor Association have collaboratively organized a panel of relevant experts to develop the Guideline for diagnosis and treatment of infection after internal fixation of closed lower limb fractures in adults ( version 2025). The guideline proposed 10 recommendations, aiming to provide a foundation for standardized diagnosis and treatment of postoperative infection in adults with closed lower limb fractures.
10.Efficacy and mechanism of botulinum toxin type A combined with static progressive stretching in the treatment of traumatic knee stiffness in rats
Ke CHEN ; Xin ZHANG ; Kai REN ; Yingying LIAO ; Xin HE ; Xiaoju LI
Chinese Journal of Trauma 2025;41(2):201-211
Objective:To investigate the efficacy and mechanism of botulinum toxin type A (BTX-A) combined with static progressive stretching (SPS) in the treatment of traumatic knee stiffness in rats.Methods:Forty healthy male SD rats aged 8 weeks and weighing 220-300 g, were randomly divided into blank control group ( n=8) and model groups ( n=28) (including injury group, BTX-A group, SPS group and BTX-A+SPS group, with 7 in each group). Hlidebrand′s method was used to construct a traumatic knee stiffness model, with the following main steps: destruction of the joint capsule, Kirschner wire fixation, joint drilling, and removal of the internal fixation at 4 weeks. The blank control group did not receive any treatment and could move freely in the cage. The injury group moved freely after successful modeling. On the day of internal fixation removal, BTX-A was injected into the joint cavity in group BTX-A, SPS treatment was started in the SPS group, BTX-A was injected into the joint cavity and SPS treatment was started in the BTX-A+SPS group. The treatments lasted 16 days. The range of motion (ROM) and joint stiffness were measured before treatment and at 16 days after treatment. At 16 days after treatment, knee joint tissue was collected and the rats were sacrificed, and the articular capsule fibrous tissue proliferation was observed by HE and Masson staining. The expression levels of phosphorylated (p)-Smad2, Smad2, p-Smad3, Smad3, Smad4, transforming growth factor-β1 (TGF-β1), collagen type I, collagen type III, and α-smooth actin (α-SMA) were determined by Western blot. The ratio of phosphorylated protein to total protein was calculated to reflect the phosphorylation level. Results:(1) ROM: Before treatment, the ROM in the blank control group was significantly higher than that in the other groups ( P<0.05), with no significant difference in ROM among the other groups ( P>0.05). At 16 days after treatment, ROM in the injury group, BTX-A group, SPS group, and BTX-A+SPS group was lower than that in the blank control group ( P<0.05), among which ROM in the BTX-A+SPS group was significantly higher than that in the injury group, BTX-A group, and SPS group ( P<0.05). At 16 days after treatment, there was no significant difference in ROM before and after treatment in the blank control group ( P>0.05), and ROM in the other groups was significantly increased compared with that before treatment ( P<0.01). (2) Joint stiffness: At 16 days after treatment, the joint stiffness levels in the injury group, the BTX-A group, and the SPS group were (0.95±0.24)N·cm/°, (0.86±0.22)N·cm/°, and (0.65±0.09)N·cm/° respectively, which were significantly lower than (0.36±0.03)N·cm/° in the blank control group ( P<0.05). The joint stiffness level of the BTX-A+SPS group was (0.49±0.04)N·cm/°, which was not significantly different from that in the blank control group ( P>0.05), but was significantly lower than those in the injury group, BTX-A group, and SPS group ( P<0.05). (3) Fibrous tissue proliferation: at 16 days after treatment, the joint capsular structure in the blank control group was complete and clear, the fibers were arranged in order, and there was no obvious fibrous tissue proliferation. The pathological changes in the injury group were the most serious, with a large number of synovial fibrous tissue proliferation, significantly increased blood vessels in the tissue, and inflammatory cell infiltration. Compared with the SPS group and BTX-A group, the lesions in BTX-A+SPS group were milder, with only slight increase in the number of synovial cells but no obvious vascular proliferation or lymphocytes, and the overall lesions were the least severe. (4) Protein expression: the ratios of p-Smad2/Smad2 in the injury group, BTX-A group and SPS group were 1.552±0.234, 1.328±0.272 and 1.194±0.277 respectively, which were higher than 0.794±0.082 in the blank control group ( P<0.05). The ratio of p-Smad2/Smad2 in the BTX-A+SPS group was 1.013±0.123, which was not significantly different from those in the blank control group, BTX-A group or SPS group ( P>0.05), but was lower than that in the injury group ( P<0.05). At 16 days after treatment, the p-Smad3/Smad3 ratios in the injury group, BTX-A group, SPS group and BTX-A+SPS group were 2.272±0.309, 1.664±0.285, 1.381±0.276 and 1.003±0.060 respectively, which were higher than 0.515±0.051 in the blank control group ( P<0.05). The p-Smad3/Smad3 ratio in the BTX-A+SPS group was significantly lower than those in the injury group, BTX-A group and SPS group ( P<0.05). At 16 days after treatment, the level of Smad4 in the injury group (1.001±0.015) was higher than 0.294±0.076 in the blank control group ( P<0.05). However, there was no significant difference between the BTX-A group (0.664±0.051), SPS group (0.833±0.045), BTX-A+SPS group (0.467±0.068) or the blank control group ( P>0.05). The level of Smad4 in the BTX-A+SPS group was significantly lower than those in the injury group, BTX-A group and SPS group ( P<0.05). At 16 days after treatment, the level of TGF-β1 in the injury group (1.004±0.407) was higher than 0.269±0.122 in the blank control group ( P<0.05), while there was no significant difference between the BTX-A group (0.564±0.194), SPS group (0.422±0.086) and BTX-A+SPS group (0.347±0.161) and the blank control group ( P>0.05). The level of TGF-β1 in the BTX-A+SPS group was significantly lower than those in the injury group, BTX-A group and SPS group ( P<0.05). At 16 days after treatment, the level of type I collagen in the injury group was 0.999±0.170, higher than 0.299±0.139 in the blank control group ( P<0.05), while there was no significant difference between the BTX-A group (0.542±0.278), SPS group (0.561±0.165), and BTX-A+SPS group (0.537±0.045) and the blank control group ( P>0.05). The level of collagen type I in the BTX-A+SPS group was significantly lower than those in the injury group, BTX-A group, and SPS group ( P<0.05). At 16 days after treatment, the level of type III collagen in the injury group was 1.002±0.126, higher than 0.239±0.106 in the blank control group ( P<0.05), while there was no significant difference between the BTX-A group (0.661±0.062), SPS group (0.595±0.062), and BTX-A+SPS group (0.504±0.269) and the blank control group ( P>0.05). The level of collagen type III in the BTX-A+SPS group was significantly lower than those in the injury group, BTX-A group, and SPS group ( P<0.05). At 16 days after treatment, the level of α-SMA in the injury group was 0.998±0.074, higher than 0.130±0.023 in the blank control group ( P<0.05), while there was no significant difference between the BTX-A group (0.358±0.060), SPS group (0.432±0.230), and BTX-A+SPS group (0.293±0.135) and the blank control group ( P>0.05). The level of α-SMA in the BTX-A+SPS group was significantly lower than those in the injury group, BTX-A group and SPS group ( P<0.05). Conclusions:Compared with single treatment, the combination of BTX-A and SPS demonstrates significantly greater efficacy in the treatment of traumatic knee stiffness in rats. This combined approach not only enhances joint mobility and elasticity but also effectively inhibits joint capsule fibrosis. The underlying mechanism may involve the further suppression of TGF-β1 expression in the joint capsule, leading to reduced phosphorylation levels of Smad2 and Smad3. This, in turn, inhibits the binding of Smad2 and Smad3 to the Smad4 receptor, ultimately downregulating the expression of the downstream proteins of the TGF-β/Smad signaling pathway, such as collagen type I, collagen type III and α-SMA.


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