1.Huoxue Wentong Prescription Ameliorates Post-myocardial Infarction Ventricular Remodeling in Rats via HIF-1α/TGF-β1/POSTN-mediated Regulation of EndMT
Xiaoning SUN ; Xiaoxiao ZHANG ; Cheng LI ; Qingqing WANG ; Jianguo LIN ; Mingyue WANG ; Ying YANG ; Kuiwu YAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(22):148-158
ObjectiveTo explore the potential mechanism by which Huoxue Wentong prescription (HXWT) ameliorates post-myocardial infarction (MI) ventricular remodeling by regulating endothelial-to-mesenchymal transition (EndMT) via the hypoxia-inducible factor-1α (HIF-1α)/transforming growth factor-β1 (TGF-β1)/periostin (POSTN) signaling pathway. MethodsEighty SPF-grade male Sprague-Dawley rats were randomly assigned to a sham-operated group (n=10) and a modeling group (n=70). The MI model was established by left anterior descending coronary artery ligation. Successfully modeled rats were further randomized into a model group, a sacubitril/valsartan (Entresto, 0.021 g·kg-1) group, and low, medium, and high-dose (4.15, 8.30, and 16.60 g·kg-1, respectively) HXWTF groups (n=10 per group). From postoperative day 2, rats received HXWTF or sacubitril/valsartan by gavage once daily for 4 weeks, and those in sham and model groups received an equal volume of normal saline. After intervention, general conditions were assessed. Echocardiography was performed to measure the left ventricular ejection fraction (LVEF), left ventricular fractional shortening (LVFS), left ventricular end diastolic diameter (LVEDD), and left ventricular end systolic diameter (LVESD). Hematoxylin and eosin (HE) staining was employed to evaluate myocardial histopathology, and Masson's trichrome staining to assess myocardial fibrosis. Serum levels of B-type natriuretic peptide (BNP), interleukin (IL)-6, IL-10, and matrix metalloproteinase-9 (MMP-9) were determined by enzyme-linked immunosorbent assay (ELISA). Collagen type Ⅰ (ColⅠ) expression was assessed by immunohistochemistry. Immunofluorescence double staining was adopted to detect the expression of platelet endothelial cell adhesion molecule-1 (CD31) and α-smooth muscle actin (α-SMA). Western blot was performed to quantify the expression of collagen type Ⅰ α2 (Col1A2), VE-cadherin, CD31, α-SMA, HIF-1α, TGF-β1, and POSTN. ResultsCompared with the sham group, the model group exhibited thinning and collapse of the left ventricular wall, decreased LVEF and LVFS and increased LVEDD and LVESD (P<0.01), disrupted myocardial fibers with marked inflammatory cell infiltration, increased collagen deposition, enhanced ColⅠ immunoreactivity, elevate serum BNP, IL-6, and MMP-9 levels, a reduced IL-10 level (P<0.01), downregulated expression of CD31 and VE-cadherin, and upregulated expression of α-SMA, Col1A2, HIF-1α, TGF-β1, and POSTN (P<0.05,P<0.01), accompanied by increased CD31/α-SMA co-localization. Compared with the model group, the sacubitril/valsartan group and all HXWTF groups showed improved ventricular wall structure, increased LVEF and LVFS, decreased LVEDD and LVESD, attenuated myocardial injury with reduced inflammatory infiltration and collagen deposition, declined serum BNP, IL-6, and MMP-9 levels, an elevated serum IL-10 level, upregulated expression of CD31 and VE-cadherin, downregulated expression of α-SMA, Col1A2, HIF-1α, TGF-β1, and POSTN, and reduced CD31/α-SMA co-localization (P<0.05,P<0.01). No significant differences were observed between the sacubitril/valsartan group and the medium- and high-dose HXWTF groups. ConclusionHXWTF may regulate the HIF-1α/TGF-β1/POSTN pathway to inhibit EndMT, thereby ameliorating post-MI ventricular remodeling.
2.Study on the relationship between serum GLP-1,MCP-1,IGFBP-3 and glycolipid metabolism,bone metabolism and microvascular complications in children with T1DM
Xiaoning GUAN ; Lihua WANG ; Limin HU ; Jinfeng REN ; Wangye LI ; Jing WU ; Xiangfen YANG
International Journal of Laboratory Medicine 2025;46(14):1702-1708
Objective To investigate the relationship between serum glucagon-like peptide-1(GLP-1),monocyte chemoattractant protein-1(MCP-1),insulin-like growth factor binding protein-3(IGFBP-3)and glycolipid metabolism,bone metabolism and microvascular complications(MC)in children with type 1 diabe-tes mellitus(T1DM).Methods A total of 211 children with T1DM(T1DM group)admitted to Handan Cen-tral Hospital,Xingtai Traditional Chinese Medicine Hospital,Baoding First Central Hospital and Handan Ma-ternal and Child Health Hospital from January 2021 to February 2023 were selected,patients were divided into MC group(63 cases)and non-MC group(148 cases)according to whether MC was complicated within 1 year,and 108 healthy children who underwent physical examination during the same period were selected as control group.The levels of serum GLP-1,MCP-1,IGFBP-3 and glucose and lipid metabolism indexes[fasting plasma glucose(FPG),fasting insulin(FINS),glycosylated hemoglobin(HbA1c),homeostasis model assessment of insulin resistance(HOMA-IR),total cholesterol(TC),triglyceride(TG),high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL-C)]and bone metabolism indexes[bone specific alkaline phosphatase(BALP),osteocalcin(OST),type I collagen cross-linked C-terminal peptide(CTX)]were detec-ted.The correlation between serum GLP-1,MCP-1,IGFBP-3 and glucose and lipid metabolism,bone metabo-lism in children with T1DM were analyzed by Pearson and Spearman correlation coefficient.Taking MC in children with T1DM as the dependent variable,the influencing factors were determined by multivariate uncon-ditional Logistic regression model,and the predictive value of serum GLP-1,MCP-1 and IGFBP-3 for MC were analyzed by receiver operating characteristic curve.Results The levels of serum GLP-1,FINS,HDL-C,BALP,OST and CTX in the T1DM group were lower than those in the control group,while the levels of MCP-1,IGFBP-3,FPG,HbA1c,HOMA-IR,TG and LDL-C in the T1DM group were higher than those in the control group,the differences were statistically significant(P<0.05).Serum GLP-1 in children with T1DM was negatively correlated with FPG,HbA1c,HOMA-IR,TG and LDL-C,and positively correlated with FINS,HDL-C,BALP,OST and CTX(P<0.05).MCP-1 and IGFBP-3 were positively correlated with FPG,HbA1c,HOMA-IR,TG and LDL-C,and negatively correlated with FINS,HDL-C,BALP,OST and CTX(P<0.05).Follow-up for 1 year,the incidence of MC in 211 children with T1DM was 29.86%(63/211).Elevated HbA1c,HOMA-IR,LDL-C,MCP-1 and IGFBP-3 were independent risk factors for MC in children with T1DM,and elevated GLP-1 was an independent protective factor(P<0.05).The area under the curve of ser-um GLP-1,MCP-1 and IGFBP-3 combined to predict MC in children with T1DM was 0.919,which was grea-ter than 0.781,0.788 and 0.794 predicted by serum GLP-1,MCP-1 and IGFBP-3 alone(P<0.05).Conclu-sion The decrease of serum GLP-1 level and the increase of MCP-1 and IGFBP-3 levels are related to glyco-lipid metabolism,bone metabolism disorder and MC in children with T1DM,the combined application of ser-um GLP-1,MCP-1 and IGFBP-3 has a good predictive value for MC in children with T1DM.
3.Hyperparathyroidism-jaw tumor syndrome: a case report
Lanning JIA ; Ruoyu JIANG ; Xiaoning WANG ; Ke ZHAO ; Fuxin LI ; Yizeng WANG ; Xianghui HE
Chinese Journal of Endocrine Surgery 2025;19(5):787-789
Hyperparathyroidism-jaw tumor syndrome (HPT-JT) is a rare familial autosomal dominant genetic disease with primary hyperparathyroidism, jaw tumors, kidney tumors and uterine tumors caused by cell division cycle 73 (CDC73) germline mutations. A 42-year-old male patient was admitted for pancreatitis and further examination revealed elevated PTH at 54.00pmol/L and a history of jaw tumors. This patient was diagnosed as HPT-JT finally and underwent upper right, lower right, and upper left parathyroid glands resection and genetic testing. Postoperative pathology revealed that atypical adenomatous nodules of parathyroid glands with extensive atypia and nucleus division and parathyroid hyperplasia and whole exome sequencing suggested that the CDC73 mutation.
4.Application of the Anderson sampler in the inspection for the filtration efficiency for bacteria in medical mask
Di LEI ; Chen WANG ; Minjuan ZHANG ; Cunlin LONG ; Jian REN ; Zhijie ZHAO ; Yuwei LI ; Yun LING ; Xiaoning SUN ; Jing ZHAO
China Medical Equipment 2025;22(3):160-163
The medical mask,which is used as an important tool of preventing the spread of respiratory diseases,can effectively block the transmission of biological aerosols.The detection for the filtration efficiency of bacteria in medical mask is particular importance.The Andersen sampler,is one kind of device that samples microbial aerosols,is widely used in the inspection for the filtration efficiency for bacteria in medical masks.It mainly consists of six impactors with different pore sizes.It simulates the deposition process of the most of particles at different positions in respiratory system through the bacterial particles in biological aerosols impact respectively the surface of petri dishes with agar under different pore sizes.This paper explored the development background,structure and sampling principle,operation and counting procedures of the Andersen sampler,as well as its application and importance in the inspection for the filtration efficiency for bacteria in medical mask.
5.Study on the Characteristics of TCM Syndromes of Insomnia Based on Multiple Data Analysis Methods
Xianbei WANG ; Rui ZHANG ; Jiwei ZHANG ; Yuying XU ; Ning SUN ; Miaoran WANG ; Xiaoning TAN ; Yufei WU ; Zirong LI ; Jing CAO ; Taiwei LOU ; Rui WEI ; Hongjin DU ; Qiuyan LI
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(5):25-31
Objective To explore the distribution of common TCM syndromes and symptoms of insomnia;To prepare for the construction of the theoretical framework and item pool of syndrome diagnosis and efficacy evaluation scale.Methods TCM guideline standards of insomnia,textbooks and journals over the years were retrieved,the information of TCM syndromes,syndrome elements and symptoms was extracted,the guideline textbook and journal database were established,and descriptive statistics,association rules,systematic clustering,factor analysis,potential categories and implicit structure analysis were carried out.Results Totally 116 guide standards and textbooks over the years were included,and 454 articles of journals were included.The high-frequency symptoms accounted for≥3%of the guide textbooks and journal databases were 87 and 79 categories,respectively,and the cumulative proportion was 87.48%and 87.75%,respectively.According to the analysis results,five common TCM syndromes and their characteristic symptom classification of insomnia were finally deduced.According to the frequency/person time distribution,they were heart and spleen deficiency syndrome,yin deficiency and fire hyperactivity syndrome,liver fire disturbing heart syndrome,phlegm heat disturbing heart syndrome,heart and gallbladder qi deficiency syndrome.Conclusion There are five common TCM syndromes of insomnia,and the characteristic symptoms of each TCM syndrome provide a reference source for the theoretical framework of syndrome diagnosis and efficacy evaluation scale and the establishment of item pool.
6.Evaluation of the Efficacy of Kai-Xin-San Combined with Fluoxetine in Improving Intestinal Absorption Damage in Depres-sion Model Mice
Xin LI ; Xuan LI ; Xiaoning HUANG ; Lingxin HUANG ; Jiahui WU ; Tingxia DONG ; TSIM Wah KEUNG ; Jin'ao DUAN ; Yue ZHU
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(3):313-322
OBJECTIVE To evaluate the efficacy and mechanism of Kai-Xin-San in improving intestinal function damage in-duced by intragastric administration of fluoxetine in chronic unpredictable mild stress(CUMS)depression model mice.METHODS A CUMS depression mouse model was established and treated with fluoxetine(9 mg·kg-1·d-1),low-dose(1.5 g·kg-1·d-1)and high-dose(4.5 g·kg-1·d-1)Kai-Xin-San,fluoxetine combined with low-dose(9 mg·kg-1·d-1+1.5 g·kg-1·d-1)and high-dose(9 mg·kg-1·d-1+4.5 g·kg-1·d-1)Kai-Xin-San,and mosapride combined with fluoxetine(2 mg·kg-1·d-1+9 mg·kg-1·d-1)for 28 consecutive days.The body weight of mice was measured;the food utilization was calculated and the serum D-xylose content was measured to evaluate the intestinal mucosal absorption capacity of mice;HE staining was used to evaluate the intestinal structural dam-age of mice;TUNEL staining was used to evaluate the intestinal tissue apoptosis of mice;ELISA was used to detect the expression lev-els of brain gut peptides such as vasoactive intestinal peptide(VIP),gastrin(MTL),substance P(SP)and Ghrelin in the intestine of mice;Western blot was used to detect the expression of apoptosis signaling pathway proteins.RESULTS Compared with the model group,fluoxetine significantly reduced the body weight of mice after 2 weeks of administration(P<0.05);the food utilization and ser-um D-xylose content of mice were significantly reduced after 4 weeks of administration(P<0.05),and the intestinal villi of depressed mice were damaged(P<0.05)and intestinal epithelial apoptosis of mice was enhanced(P<0.01);the expression of VIP in the small intestine of mice was upregulated(P<0.05),and the expression of MTL,SP and Ghrelin was downregulated(P<0.05,P<0.01);cleaved Caspase-3/Caspase-3 and cleaved Caspase-9/Caspase-9 in the intestinal apoptosis signaling pathway of mice were upregulat-ed(P<0.05,P<0.01).Compared with the fluoxetine group,the body weight of mice was significantly increased after 2 weeks of com-bined use of Kai-Xin-San and fluoxetine(P<0.05,P<0.01).After 4 weeks of combined use of high-dose Kai-Xin-San and fluoxe-tine,the food utilization and serum D-xylose expression of mice were significantly increased(P<0.05);intestinal villus damage was improved(P<0.05);intestinal epithelial tissue apoptosis was significantly reduced(P<0.01);small intestinal VIP expression was significantly downregulated(P<0.01),and the expression of MTL,SP and Ghrelin was significantly upregulated(P<0.05);cleaved Caspase-3/Caspase-3 and cleaved Caspase-9/Caspase-9 in the apoptosis signaling pathway were significantly reduced(P<0.05,P<0.01).CONCLUSION Kai-Xin-San has the effect of improving the gastrointestinal motility and intestinal absorption function dam-age caused by fluoxetine in depressed mice.Its mechanism may be related to improving the expression of brain gut peptide in the small intestine and inhibiting intestinal villi damage and intestinal tissue apoptosis.
7.Evaluation of the Effect of Chinese Medicine Formula Kai-Xin-San Combined with Fluoxetine on Hippocampal Neural Stem Cells in Chronic Stress Induced Depression Model Mice
Lingxin HUANG ; Xin LI ; Lei YUAN ; Yun ZHU ; Xiaoning HUANG ; Xuan LI ; Huaqiang ZHAN ; Jinao DUAN ; Lejun LI ; Yue ZHU
World Science and Technology-Modernization of Traditional Chinese Medicine 2025;27(4):1035-1046
Objective To evaluate the effect of Kaixin San(KXS)combined with fluoxetine on hippocampal neural stem cells in mice with chronic stress stress and depression.Methods A mouse model of depression was constructed using the method of chronic unpredictable stress stress,and the highest dose of KXS water extract and fluoxetine for clinical application was given for 28 days,and behavioral tests were carried out.Nissl staining was used to detect the pathological status of hippocampal tissues in mice.The expression of TUNEL and Nestin in mouse hippocampus was determined by immunofluorescence.Western blotting was used to detect the expressions of apoptotic proteins cleaved caspase-3 and caspase-3,pyroptosis proteins GSDMD and cleaved caspase-1,as well as the expression of neural stem cell marker Nestin in the hippocampus,and the expression of Wnt/β-catenin signaling pathway-related proteins in the hippocampus.Results The combination of KXS extract and fluoxetine can significantly improve the depression-like behavior of model mice,and the effect is better than fluoxetine alone.The combination inhibited the activation of apoptosis and pyroptosis signaling pathways in the hippocampus when used alone with high-dose fluoxetine,significantly upregulated the expression of Nestin,and regulated the expression of Wnt/β-catenin signaling pathway protein.Conclusion The combination of KXS and high-dose fluoxetine can improve apoptosis and pyroptosis in the hippocampus of stress stress and depression model mice,and upregulate the expression of neural stem cell marker Nestin by regulating the Wnt/β-catenin signaling pathway,which may be a key link to improve the antidepressant effect of the combination drug.
8.Performance verification of fully automated chemiluminescence immunoassay analyzer in measuring special sequence indicators of serum β-CTx
Di LEI ; Jian REN ; Minjuan ZHANG ; Xiaoning SUN ; Yingjun LI ; Xiaodong ZHANG ; Cunlin LONG
China Medical Equipment 2025;22(8):57-60
Objective:To verify the performance of MAGLUMI 4000 fully automatic chemiluminescence immunoassay analyzer in measuring special sequence of β-Collagen(β-CTx).Methods:Referring to a series of standards included WS/T 492-2016"Verification of performance for precision and trueness of quantitative measurements in clinical laboratories"and CNAS-GL037 2019"Guidance on the verification of quantitative measurement procedures used in the clinical chemistry",the precision,trueness,and linear interval of MAGLUMI 4000 fully automatic chemiluminescence immunoassay analyzer were verified in measuring β-CTx.Results:The intra batch precisions(repeatability)of MAGLUMI 4000 fully automatic chemiluminescence immunoassay analyzer were respectively 3.22%and 3.49%in measuring serum β-CTx samples with low and high values.The intermediate precisions(precision within laboratory)were respectively 4.35%and 3.29%,both of which met the requirements of laboratory.The results of trueness verification showed that the bias of samples with low concentration was-2.4%,and the bias of samples with high concentration was-2.1%.The expected values of the standards with low and high values were all between the corresponding up and low validation limits of them,which met the judgment criteria.The linear interval was 0.03-6.00 ng/mL,which was within the linear interval,and it can meet the requirements of manufacturer′s claim.Conclusion:The precision,trueness and linear interval of MAGLUMI 4000 fully automatic chemiluminescence immunoassay analyzer all passed the verification in measuring β-CTx,which indicates the performance of the project can meet the quality specifications.
9.Guideline for the prevention of intraoperative acquired pressure injury in paraplegic patients with spinal cord injury (version 2025)
Aijun XU ; Shuixia LI ; Bo CHEN ; Mengyuan YE ; Lejiao LANG ; Ning NING ; Lin ZHANG ; Changqing LIU ; Zhonglan CHEN ; Weihu MA ; Weishi LI ; Xiaoning WANG ; Dongmei BIAN ; Jiancheng ZENG ; Xin WANG ; Yuan GAO ; Yaping CHEN ; Jiali CHEN ; Yun HAN ; Xiuting LI ; Yang ZHOU ; Xiaojing SU ; Qiong ZHANG ; Tianwen HUANG ; Ping ZHANG ; Hua LIN ; Xingling XIAO ; Ruifeng XU ; Fanghui DONG ; Bing HAN ; Luo FAN ; Yanling PEI ; Suyun LI ; Xiaoju TAN ; Rongchen GUO ; Yefang ZOU ; Xiaoyun HAN ; Junqin DING ; Yi WANG ; Shuhua DENG ; Jinli GUO ; Yinhua LIANG ; Yuan CEN ; Xiaoqin LIU ; Junru CHEN ; Haiyang YU ; Lunlan LI ; Ying REN ; Yunxia LI ; Jianli LU ; Ying YING ; Lan WEI ; Yin WANG ; Qinhong XU ; Yanqin ZHANG ; Yang LYU ; Shijun ZHANG ; Sui WENJIE ; Sanlian HU ; Shuhong YANG ; Guoqing LI ; Jingjing AN ; Baorong HE ; Leling FENG
Chinese Journal of Trauma 2025;41(6):530-541
Paraplegia caused by spinal cord injury is a serious neurological complication, for which surgery is currently the main treatment method. Due to different surgical approaches, patients are usually expected to maintain a passive prone position for a long time or switch between the supine and prone positions. Affected by multiple factors such as neurogenic sensory disorders, pathological changes in muscle tone and operative duration, the risk of intraoperative acquired pressure injury (IAPI) is significantly increased. Current clinical prevention strategies for IAPI in these patients predominantly focus on localized pressure relief during positioning, lacking systematic, standardized comprehensive prevention protocols or evidence-based guidelines. To address it, Department of Nursing, Orthopedics Branch, China International Exchange and Promotive Association for Medical and Health Care, Spinal Trauma Professional Committee, Orthopedics Branch, Chinese Medical Doctor Association, Nursing Group of Spine and Spinal Cord Professional Committee of Chinese Association of Rehabilitation Medicine organized experts in relevant fields to formulate Guideline for the prevention of intraoperative acquired pressure injury in paraplegic patients with spinal cord injury ( version 2025), based on evidence-based medical evidence and latest research results and clinical practice at home and abroad. Eleven recommendations were put forward from the aspects of preoperative risk assessment, intraoperative prevention strategies, postoperative handover and monitoring, and supportive mechanisms for IAPI prevention, aiming to standardize the prevention measures and management strategies of IAPI in paraplegic patients with spinal cord injury and accelerate the recovery of patients and improve the therapeutic effect.
10.Inhibition of osteogenic differentiation of mouse bone marrow mesenchymal stem cells and maxillary expansion osteogenesis by cytoskeleton-associated protein 4 knockout
Hanzhe WANG ; Dihao TAO ; Shiping CHANG ; Xiaoning HE ; Bei LI ; Yimin ZHAO
Chinese Journal of Stomatology 2025;60(5):525-533
Objective:To investigate the effect of cytoskeleton-associated protein 4 (CKAP4) gene knockout on maxillary expansion osteogenesis and its regulatory mechanism on the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSC).Methods:Nineteen wild type (WT) and nineteen CKAP4 gene knockout (Ckap4 -/-) mice aged 6-8 weeks were selected to establish a mouse model of rapid maxillary expansion. Samples were taken on the 7th and 14th day after the operation. Micro-CT and HE staining were used to evaluate bone regeneration. Tissue proteins in the modeled area were collected, and Western blotting analysis (WB) was used to detect the protein expression levels of alkaline phosphatase (ALP), Runt-related transcription factor 2 (RUNX2), and osteocalcin (OCN). BMSC were isolated from WT and Ckap4 -/- mice. The expression of surface markers CD29, Sca-1, CD44, CD45, CD34, and CD11b was detected by flow cytometry, and cell proliferation ability was detected by 5-ethynyl-2'-deoxyuridine (EdU). After 7 days of osteogenic induction, real-time fluorescence quantitative PCR (RT-qPCR) and WB were used to detect the expression levels of RUXN2, ALP, OCN, protein kinase B (AKT), and phosphorylated protein kinase B (p-AKT). After 21 days, alizarin red staining and cetyl pyridine chloride quantification were used to detect the differences in mineralized nodule formation in each group. In CKAP4 gene knockout BMSC, the small-molecule AKT agonist sc79 (4 μg/ml) was added as the intervention group (Ckap4 -/- +sc79), and dimethyl sulfoxide (DMSO) treatment was used as the control group (Ckap4 -/- +DMSO). After osteogenic induction, RT-qPCR, WB, and alizarin red staining were used to compare the osteogenic differentiation differences between the two groups of cells. Results:The micro-CT results showed that at 7 days and 14 days after surgery, the new bone volume in the Ckap4 -/- group [(0.070±0.010) and (0.146±0.019) mm 3] was significantly lower than that in the WT group [(0.094±0.006) and (0.196±0.013) mm 3] (both P<0.01). HE-stained histological sections showed that the area of new bone tissue in the Ckap4 -/- group at 7 days and 14 days after surgery [(0.101±0.008) and (0.158±0.010) mm 2] was also significantly lower than that in the WT group [(0.116±0.005) and (0.183±0.008) mm 2] (both P<0.05). WB was used to detect the tissue proteins in the maxillary modeling area of mice in the two groups 7 days after surgery. The results showed that the expression levels of ALP, RUNX2 and OCN in the Ckap4 -/- group were significantly lower than those in the WT group. BMSC from wild-type mice and CKAP4 knockout mice were both positively expressed for CD29, CD44, and Sca-1, and basically not expressed for CD45, CD34, and CD11b. EdU assay showed that there was no significant difference in the proliferation ability of cells in the two groups. After 21 days of osteogenic induction of BMSC, alizarin red staining results showed that the number of mineralized nodules in the Ckap4 -/- group was significantly less than that in the WT group. After adding sc79, the number of mineralized nodules increased significantly, which was consistent with the results of cetyl pyridine chloride quantification. After 7 days of osteogenic induction, It was found that the expression levels of ALP, RUNX2, and OCN in the CKAP4 -/-group (0.751±0.066, 0.484±0.040, 0.679±0.063) were significantly lower than those in the WT group (1.000±0.113, 1.000±0.081, 1.000±0.113) (all P<0.001). The results of WB were consistent with those of RT-qPCR. At the same time, the WB results showed that the level of p-AKT protein in the CKAP4 -/-group (0.518±0.114) was significantly lower than that in the WT group (1.000±0.234) ( P<0.05). After treatment with sc79 for 7 days of osteogenic induction, RT-qPCR was used to detect the gene expression levels of ALP, RUNX2, and OCN. The results showed that the expression levels in the CKAP4 -/-+sc79 group (2.755±0.353, 4.800±0.990, 2.524±0.137) were significantly higher than those in the CKAP4 -/-+DMSO group (1.000±0.078, 1.000±0.247, 1.000±0.175) (all P<0.001). Conclusions:CKAP4 knockout inhibits the osteogenic differentiation of BMSC by reducing the activity of the PI3K/AKT signaling pathway, thereby suppressing osteogenesis in maxillary expansion.

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