1.Mechanism of Icariin in Regulating TGF-β1/Smad Pathway to Induce Autophagy in Human Bone Microvascular Endothelial Cells
Yaqi ZHANG ; Yankun JIANG ; Guoyuan SUN ; Bo LI ; Ran DING ; Cheng HUANG ; Weiguo WANG ; Qidong ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):123-130
ObjectiveTo investigate the regulatory effect of icariin (ICA) on transforming growth factor-β1 (TGF-β1)/Smad pathway in bone microvascular endothelial cells (BMECs) and the effect on autophagy in BMECs. MethodsBMECs were isolated and cultured, and the cell types were identified by immunofluorescence. Cells were divided into the control group, model group (0.1 g·L-1 methyl prednisolone), ICA group (0.1 g·L-1 methyl prednisolone +1×10-5 mol·L-1 ICA), and TGF-β inhibitor group (0.1 g·L-1 methyl prednisolone +1×10-5 mol·L-1 ICA +1×10-5 mol·L-1 LY2157299). Transmission electron microscopy was used to observe the ultrastructure and autophagosome number of BMECs. Autophagy double-standard adenovirus was used to monitor the confocal autophagy flow generation of each cell. Real-time quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to detect the gene and protein expression of autophagy in the TGF-β1/ Smad pathway. ResultsAfter cell separation culture, platelet endothelial cell adhesion molecule (CD31) and von willebrand factor (vWF) immunofluorescence identified BMECs. Transmission electron microscopy showed that the cell membrane was damaged, and the nucleus was pyknotic and broken in the model group. Compared with the model group, the ICA group had complete cell membranes, clear structures, with autophagy-lysosome sparsely distributed. The confocal photo showed that BMECs had autophagosomes and autophagy-lysosomes, and the autophagy expression of the ICA group was similar to that of the blank group. Compared with the blank group, in the model group and the LY2157299 group, autophagosomes and autophagy-lysosomes were barely seen in the autophagy flow. Compared with the blank group, the mRNA and protein expressions of autophagy effector protein 1 (Beclin1) and microtubule-associated protein 1 light chain 3B (LC3B) in the model group were significantly decreased (P<0.01), and those of ubiquitin-binding protein (p62) were significantly increased (P<0.01). The mRNA expression of TGF-β1, Smad homolog 2 (Smad2), and Smad homolog 3 (Smad3) decreased (P<0.05, P<0.01). The protein expressions of TGF-β1, p-Smad2, and p-Smad3 were significantly decreased (P<0.01). Compared with those of the model group, the mRNA and protein expression of Beclin1 and LC3B in BMECs of the ICA group increased (P<0.01), and those of p62 significantly reduced (P<0.01). The mRNA expression of TGF-β1, Smad2, and Smad3 increased significantly (P<0.01). The protein expression of TGF-β1, p-Smad2, and p-Smad3 increased significantly (P<0.01). Compared with those in the model group, the mRNA and protein expressions of Beclin1, LC3B, and p62 in the inhibitor group were not statistically significant. The expression of key genes and proteins of the TGF-β1 pathway in the inhibitor group was not statistically significant. ConclusionICA can promote glucocorticoid-induced autophagy expression of BMECs, and its mechanism may be related to activating the TGF-β1/Smad signaling pathway.
2.Effect of Icariin on Steroid-induced Ferroptosis in Rat Bone Microvascular Endothelial Cells
Jiancheng TANG ; Yue ZHANG ; Ruichen JIANG ; Zhengrong YUE ; Ming LI ; Yaqi ZHANG ; Zetao YIN ; Weiguo WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):131-140
ObjectiveTo investigate the effect of icariin (ICA) on steroid-induced ferroptosis in bone microvascular endothelial cells (BMECs). MethodsRat BMECs were selected and treated with 500 mg·L-1 hydrocortisone for 1.5 h to establish a ferroptosis model of BMECs. The experimental cells were divided into a blank group, hormone group (500 mg·L-1 hydrocortisone), ICA group (500 mg·L-1 hydrocortisone + 34 mg·L-1 ICA), and ferroptosis agonist group (500 mg·L-1 hydrocortisone + 34 mg·L-1 ICA + 2.7 mg·L-1 erastin). Cell viability was detected by CCK-8. The levels of ferrous ion, glutathione (GSH), malondialdehyde (MDA), superoxide dismutase (SOD), and reactive oxygen species (ROS) were detected by related kit species. The ferroptosis-related proteins, such as glutathione peroxidase 4(GPX4), ferritin light chain (FTL), and transferrin receptor protein1 (sTfR) were detected by Western blot, as well as autophagy-related proteins including microtubule-associated protein 1 light chain 3B (LC3B), Beclin1, B-cell lymphoma-2 (Bcl-2), and Caspase-3. Results500 mg·L-1 hydrocortisone intervention for 1.5 h could effectively induce ferroptosis in BMECs, and ferroptosis levels could reach a peak as the intervention continued. In terms of cellular antioxidant capacity, compared with those in the blank group, the cell vitality, GSH in the hormone group decreased significantly, and the levels of ROS, SOD, MDA, and ferrous ions were significantly increased (P<0.01). Compared with those in the hormone group, the cell viability, GSH were significantly increased, and the levels of ROS, SOD, MDA, and ferrous ions were decreased in the ICA group (P<0.01). Compared with those in the ICA group, the cell vitality, GSH in the ferroptosis agonist group decreased significantly, and the levels of ROS, SOD, MDA, and ferrous ions increased significantly (P<0.01). In terms of the relationship between ferroptosis and autophagy, compared with the blank group, the hormone group had significantly increased expression levels of LC3B, sTfR, Beclin1, and FTL and significantly decreased expression levels of GPX4 (P<0.01). Compared with the hormone group, The ICA group had significantly decreased expression levels of LC3B, sTfR, and FTL and significantly increased expression levels of Beclin 1 and GPX4 (P<0.01). Compared with those in the ICA group, the expression levels of LC3B, sTfR, and FTL increased in the rapamycin group, and those of Beclin 1 and GPX4 decreased (P<0.01). In terms of cell ferroptosis and apoptosis,compared with the blank group, the hormone group had significantly increased expression levels of FTL, sTfR and Caspase-3 and significantly decreased expression levels of GPX4, and Bcl-2 (P<0.01). Compared with the hormone group, the ICA group had significantly decreased expression levels of FTL, sTfR and Caspase-3 and significantly increased expression levels of GPX4, and Bcl-2 (P<0.01). Compared with those in the ICA group, the expression levels of FTL, sTfR and Caspase-3 in the ferroptosis agonist group were increased, and the expression levels of GPX4, and Bcl-2 were decreased (P<0.01). In terms of cell function,compared with that in the blank group, the ability of cell migration and tube formation was significantly decreased in the hormone group (P<0.01). Compared with that in the hormone group, the cell migration and tube formation ability in the ICA group were significantly increased (P<0.01). ConclusionFerroptosis is involved in steroid-induced damage in BMECs. ICA can inhibit steroid-induced ferroptosis in BMECs, and the mechanism may be associated with the inhibition of ferroptosis by regulating autophagy.
3.Mechanism of Traditional Chinese Medicine in Treating Steroid-Induced Osteonecrosis of Femoral Head via Regulating PI3K/Akt Pathway: A Review
Yaqi ZHANG ; Bo LI ; Jiancheng TANG ; Ran DING ; Cheng HUANG ; Yaping XU ; Qidong ZHANG ; Weiguo WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):141-149
Steroid-induced osteonecrosis of the femoral head (SONFH) is a severe musculoskeletal disorder often induced by the prolonged or excessive use of glucocorticoids. Characterized by ischemia of bone cells, necrosis, and trabecular fractures, SONFH is accompanied by pain, femoral head collapse, and joint dysfunction, which can lead to disability in severe cases. The pathogenesis of SONFH involves hormone-induced osteoblast apoptosis, bone microvascular endothelial cell (BMEC) apoptosis, oxidative stress, and inflammatory responses. The phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling pathway plays a pivotal role in the development of the disease. Modulating the PI3K/Akt signaling pathway can promote Akt phosphorylation, thereby stimulating the osteogenic differentiation of bone marrow mesenchymal stem cells and osteoblasts, promoting angiogenesis in BMECs, and inhibiting osteoclastogenesis. The research on the treatment of SONFH with traditional Chinese medicine (TCM) has gained increasing attention. Recent studies have shown that TCM monomers and compounds have potential therapeutic effect on SONFH by intervening in the PI3K/Akt signaling pathway. These studies not only provide a scientific basis for the application of TCM in the treatment of SONFH but also offer new ideas for the development of new therapeutic strategies. This review summarized the progress in Chinese and international research on the PI3K/Akt signaling pathway in SONFH over the past five years. It involved the composition and transmission mechanisms of the signaling pathway, as well as its regulatory effects on osteoblasts, mesenchymal stem cells, osteoclasts, BMECs, and other cells. Additionally, the review explored the TCM understanding of SONFH and the application of TCM monomers and compounds in the intervention of the PI3K/Akt pathway. By systematically analyzing and organizing these research findings, this article aimed to provide references and point out directions for the clinical prevention and treatment of SONFH and promote further development of TCM in this field. With in-depth research on the PI3K/Akt pathway and the modern application of TCM, it is expected to bring safer and more effective treatment options for patients with SONFH.
4.Depressive symptoms and associated factors among middle school and college students from 2021 to 2023 in Hunan Province
Chinese Journal of School Health 2025;46(1):96-101
Objective:
To investigate the current status and trends of depressive symptoms among middle school and college students in Hunan Province, and to explore the primary related factors of depressive symptoms, so as to provide a scientific basis for strengthening mental health among students.
Methods:
A total of 279 382 students in Hunan Province were selected through a stratified cluster random sampling method from 2021 to 2023. National Survey Questionnaire on Common Diseases and Health Influencing Factors among Students was adopted for the survey, and the Center for Epidemiological Studies Depression Scale was used to assess their depressive symptoms. The χ 2 test and trend χ 2 test were used to analyze depressive symptoms prevalence and trends, and multivariable Logistic regression was used to analyze the related factors of depressive symptoms.
Results:
The prevalence of depressive symptoms among students in Hunan Province from 2021 to 2023 were 19.66%, 20.17% and 21.47%, respectively, showing an upward trend ( χ 2 trend =9.07, P <0.01). In addition, the results of the multivariable Logistic regression analysis showed that students with healthy diet ( OR=0.43, 95%CI =0.40-0.45), adequate sleep ( OR=0.88, 95%CI =0.86-0.90), and acceptable screen time ( OR=0.61, 95%CI =0.60-0.62) had lower risks in depressive symptoms detection, while students with smoking ( OR= 1.95, 95%CI =1.88-2.02), secondhand smoke exposure ( OR=1.33, 95%CI =1.30-1.36) and Internet addiction ( OR= 4.19 , 95%CI =4.05-4.34) had higher risks in depressive symptoms detection, with differences in the degree of association among different genders, educational stages and urban rural groups ( OR=0.40-6.04, Z =-12.69-11.98) ( P <0.05).
Conclusions
There is an increasing trend of depressive symptoms among middle school and college students in Hunan Province from 2021 to 2023.Targeted depression prevention measures should be taken for students with different demographic characteristics to promote their mental health.
5.Analysis on Acupuncture in the Treatment of Post-stroke Dysphagia Based on the Theory of "Latent Pathogen in Cerebral Collaterals"
Hongtao LI ; Yaqi HUANG ; Lulu YAO ; Bing GAO ; Yu YE ; Nenggui XU ; Lin WANG
Journal of Traditional Chinese Medicine 2025;66(9):901-905
Dysphagia is a common complication of stroke. Combining the principles of traditional Chinese medicine with modern research findings, it is proposed that "latent pathogen in the cerebral collaterals" constitutes the core pathogenesis of post-stroke dysphagia (PSD). In clinical practice, treatment is tailored according to the location of PSD. During the oral stage, when the pathogen invades the face and mouth, resulting in excessive salivation, acupoints are primarily selected from the foot shaoyin (少阴) kidney channel, in combination with ren mai (任脉) , du mai (督脉), chong mai (冲脉) and the spleen channel, to replenish essence and fill the marrow, dispel dampness and unblock the channels. In the pharyngeal stage, as the pathogen obstructs the throat, disrupting normal swallowing, the therapy emphasizes dredging the shaoyang (少阳) channel and warming and tonifying the jueyin (厥阴) channel, by taking acupoints mainly from the hand and foot shaoyang channels, along with the jueyin channels, so as to soothe the liver and promote bile secretion, regulate and harmonize qi and blood. During the esophageal stage, where the pathogen damages the esophagus, impeding food passage, the treatment emphasizes activating the yangming (阳明) channels and regulating taiyin (太阴) channels; acupoints are mainly selected from the foot yangming stomach channel, along with the taiyin channels, aiming to warm yang, unblock the channels and dispel stasis.
6.Study on the Pathological Mechanism-Syndrome-Treatment Patterns of Approved Chinese Patent Medicines Targeting Collateral Disorders
Pengli SU ; Peng XU ; Yanhong WANG ; Yaqi ZU ; Run YUAN ; Kun LI ; Yufeng ZHAO
Journal of Traditional Chinese Medicine 2025;66(16):1711-1718
ObjectiveTo explore the pathological mechanism-syndrome-treatment patterns of approved Chinese patent medicines (CPMs) that treat collateral disorders, providing a reference for the principle of "treating different diseases with the same therapy" in collateral pathology. MethodsCPMs that apply treatment strategies based on collateral disorders were identified from the Pharmacodia database by extracting information from the "efficacy" or "indications" sections of drug package inserts. A database was established to extract the names and compositions of the CPMs, as well as their indications, related traditional Chinese medicine (TCM) symptoms, disease locations (affected areas), and pathological factors. Frequency statistics were performed. Using the Apriori algorithm, an association rule analysis was conducted on CPMs and disease-location combinations related to the top three most frequent pathological factor combinations. Core formulas for these combinations were identified and analyzed through drug network analysis and MCODE module clustering. ResultsA total of 660 CPMs targeting collateral disorders were retrieved, involving 299 indications, 323 TCM symptoms, 21 disease locations, 19 pathological factors, and 124 pathological factor combinations. The most frequent pathological factor combinations were blood stasis (involved in 109 CPMs, 16.52%), exogenous wind (外风) -cold-dampness (involved in 43 CPMs, 6.52%), and qi deficiency-blood stasis (involved in 42 CPMs, 6.36%). Analysis of the core formulas for these combinations revealed common ingredients such as Honghua (Carthami Flos), Chuanxiong (Chuanxiong Rhizoma), Danggui (Angelicae Sinensis Radix), and Dilong (Pheretima). ConclusionCollateral disorders involve a wide range of pathogenesis and represent a fundamental mechanism in the onset and development of various diseases, characterized by obstruction and stagnation. The primary therapeutic principle is unblocking of the collaterals. Blood stasis obstructing the collaterals is the core pathological basis, and the strategy of activating blood circulation and resolving stasis to unblock the collaterals should be central to the treatment. The core medication pattern involves combining blood-activating and stasis-resolving herbs with insect-derived medicinals that unblock collaterals. Exogenous wind is often the initiating patholo-gical factor in colla-teral disorders, and the appropriate addition of wind-dispelling herbs can enrich the treatment strategies for such conditions.
7.Role of Wnt signaling Pathway in Diabetic Osteoporosis and Traditional Chinese Medicine Intervention: A Review
Yaqi ZHANG ; Weiguo WANG ; Qidong ZHANG ; Huizhong BAI ; Lingling QIN ; Feng GAO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(9):283-292
Diabetic osteoporosis (DOP) is a kind of bone complication caused by diabetes, which is characterized by the decrease of bone mineral density, the change of bone microstructure and the increase of bone fragility. The process of DOP is closely related to high glucose, insulin resistance, oxidative stress and other mechanisms. The Wnt/β-catenin signaling pathway plays an important role in mediating insulin resistance and bone metabolic balance in diabetes. Regulation of Wnt signal transduction promotes the expression of glycogen synthase kinase-3β(GSK-3β)phosphorylation and improves glucose and lipid metabolism. The Wnt/β-catenin signaling pathway is also an important way regulating osteocyte-driven bone remodeling, which not only plays an important regulatory role in the balance between osteoblasts and osteoclasts and improve bone metabolic homeostasis, but also promotes the expression of osteopontin, osteocalcin and type Ⅰ collagen, and improves bone proliferation and osteogenic differentiation by regulating the Wnt pathway. In recent years, the research of traditional Chinese medicine (TCM) in the prevention and treatment of DOP has gradually increased, and the exploration of TCM to interfere with the Wnt pathway to improve DOP has made some progress. This paper collects and summarizes the studies on the Wnt signaling pathway in glucose metabolism, bone metabolism and DOP worldwide in the past decade, as well as the related literature on the intervention of DOP by TCM compounds (classical and other compounds), single Chinese medicine and TCM monomers based on the Wnt pathway, in order to provide a reference and direction for the development of new drugs for clinical prevention and treatment of DOP.
8.Relationship between mean platelet volume/lymphocyte ratio and left atrial thrombosis in elderly patients with nonvalvular atrial fibrillation
Bin LI ; Jinchun WANG ; Zhibin WANG ; Bin ZHANG ; Yalan MAO ; Yaqi ZHANG
International Journal of Laboratory Medicine 2024;45(6):681-685
Objective To investigate the relationship between mean platelet volume/lymphocyte ratio(MPVLR)and left atrial thrombosis in elderly patients with non-valvular atrial fibrillation(NVAF).Methods A total of 178 elderly patients with NVAF admitted to the hospital from January 2019 to December 2022 were enrolled in the study.The patients were divided into thrombosis group(28 cases)and non-throm-bosis group(150 cases)according to the left atrial thrombosis judged by using esophageal echocardiography(TEE).The white blood cell count(WBC),red blood cell count(RBC),lymphocyte count,lymphocyte pro-portion,platelet count(PLT)and mean platelet volume(MPV)were detected by automatic blood cell analy-zer,and MPVLR was calculated.The liver and kidney function indicators and blood lipid indicators were detec-ted by automatic biochemical analyzer.Receiver operating characteristic(ROC)curve was used to evaluate the predictive value of MPV,lymphocyte ratio and MPVLR for left atrial thrombosis in NVAF patients.Multiva-riate Logistic regression was used to analyze the influencing factors of left atrial thrombosis in elderly NVAF patients.Results MPV,lymphocyte proportion and MPVLR in the thrombosis group were higher than those in the non-thrombosis group,and the differences were statistically significant(P<0.05).ROC curve analysis showed that the area under the curve(AUC)of MPV,lymphocyte ratio and MPVLR for predicting left atrial thrombosis in NVAF patients were 0.821(95%CI:0.764-0.882),0.771(95%CI:0.714-0.842)and 0.901(95%CI:0.861-0.949).respectively.The course of disease in the thrombosis group was longer than that in the non-thrombosis group,the proportion of patients with chronic heart failure,the proportion of patients with stroke,CH A2DS2-VASc score,LAEF,LAD,LVEDV,MPVLR,serum uric acid,MPV,lymphocyte proportion and MPVLR were higher than those in the non-thrombosis group,and LVEF was lower than that in the non-thrombosis group,the differences were statistically significant(P<0.05).Multivariate Logistic regression a-nalysis showed that disease duration ≥1.93 years(OR=3.050,95%CI:1.928-4.824),chronic heart failure(OR=3.333,95%CI:1.808-6.144),MPVLR≥3.10(OR=3.873,95%CI:1.734-8.650)were independ-ent risk factors for left atrial thrombosis in elderly NVAF patients(P<0.05).Conclusion The increase of MPVLR is associated with left atrial thrombosis in elderly patients with NVAF,and it can be used as a an in-dicator to predict left atrial thrombosis in patients with NVAF.
9.Discussion on clinical application of Dazhui acupoint based on data mining
Yaqi YAO ; Xiang ZHANG ; Yinxiang XU ; Shuxin XIONG ; Kaiqi MIAO ; Guang'an WANG
International Journal of Traditional Chinese Medicine 2024;46(2):243-248
Objective:To discuss dominant symptoms and compatibility rules of Dazhui(GV14) based on data mining.Methods:Literature related to Dazhui (GV14) was retrieved from CNKI, Wangfang, VIP, China Biomedical Literature Database (CBM) and Pubmed databases from January 1, 2012 to August 15, 2022, and the main symptoms of Dazhi (GV14) and the compatibility of acupoints were summarized. Gephi 0.9.5 software was used for complex network analysis to compare the treatment for dominant symptoms with single acupoint of Dazhi (GV14) and the compatibility of the acupoint. SPSS Modeler 18.0 software was used to analyze the association rules of acupoint combination based on Apriori algorithm. The clustering analysis of high frequency acupoints was carried out by SPSS Statistics 26.0 software.Results:A total of 722 articles were included, involving 732 prescriptions. The dominant symptoms of single acupoint were cervical spondylosis, acne, and cold; the treatment for dominant symptoms with compatibility included 14 types, such as cervical spondylosis, allergic rhinitis, ischemic stroke sequelae. The meridian compatibility was dominated by bladder meridian, and the frequency of yang meridians was higher than yin meridians. The compatibility of specific acupoints such as Xiahe acupoint, Beishu acupoint and Bahui acupoint were the main acupoints, and the high frequency acupoints were 33 acupoints such as Feishu (BL13), Baihui (GV20), Fengchi (GB20) and Zusanli (ST36), obtaining 4 series and 8 types of compatible combinations of Dazhui (GV14).Conclusions:Dazhui (GV14) is widely used in the treatment of internal diseases, such as respiratory diseases, nervous system diseases and vertebral artery type of cervical spondylosis. It tends to be flexibly used with multiple compatibility and clustering combination of specific acupoints.
10.The value of plaque-to-aorta CT value ratio in differentiating coronary lipid and fibrous plaques
Yaqi GAO ; Xuechang ZHANG ; Yao PAN ; Wei WU ; Zhaoqian WANG ; Xixia SUN ; Shuang PAN ; Hao WANG ; Zhiqiang YANG ; Chongfu JIA
Journal of Practical Radiology 2024;40(2):217-221
Objective To explore the value of plaque-to-aorta CT value ratio(standardized CT value)in differentiating coronary lipid and fibrous plaques,and to preliminarily analyze the stability of the cutoff.Methods Patients who underwent coronary computed tomography angiography(CCTA)and intravascular ultrasound(IVUS)within 1 week were included.The plaque CT value was obtained by measuring the all,four and two short-axis planes,respectively.The CT value of the ascending aorta was measured and standardized(plaque-to-aorta CT value ratio).The receiver operating characteristic(ROC)curves of the standardized and the traditional CT values were drawn.Results A total of 60 patients with 74 plaques were included,35 lipid and 39 fibrous plaques were diagnosed by IVUS.The aorta CT value was significantly correlated with the plaque(r=0.420,P<0.01);the cutoffs for the CT value of all,four and two plaque slices were 55 HU,48 HU and 52 HU,respectively,and all there of the cutoffs of standardized CT value were 0.149;the sensitivity,specificity,positive predictive value(PPV)and negative predictive value(NPV)of four-slice traditional and standardized CT values to differentiate lipid and fibrous plaques were 69%,87%,83%,76%and 91%,82%,82%,91%,respectively.Conclusion Compared with traditional CT value,the standardized CT value can greatly improve the sensitivity and NPV in differentiating coronary lipid and fibrous plaques,while maintaining modest to high specificity and PPV.Furthermore,the cutoff is stable.


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