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.Acupuncture at "four pharyngeal points" combined with Changma Xifeng tablets for simple vocal tics with liver hyperactivity disturbed wind in children: a randomized controlled trial.
Enjie WANG ; Liping LIU ; Yange WEN ; Senhui HE ; Jing LI ; Xiaojuan ZHENG ; Yaqi GENG
Chinese Acupuncture & Moxibustion 2025;45(11):1577-1581
OBJECTIVE:
To observe the effect of acupuncture at "four pharyngeal points" on simple vocal tics with liver hyperactivity disturbed wind in children.
METHODS:
Sixty children with simple vocal tics of liver hyperactivity disturbed wind were randomly divided into an observation group (30 cases, 1 case dropped out) and a control group (30 cases). The control group was given Changma Xifeng tablets orally, 3 times a day, while the observation group was treated with acupuncture at "four pharyngeal points" on the basis of the treatment in the control group, 15-20 min a time, once daily for 7 days, with a 3-day break. Both groups were treated for 3 months. The TCM syndrome score and Yale global tic severity scale (YGTSS) score of the two groups were observed before treatment and after 1, 2, 3 months of treatment, the disappearance time of simple vocal tics was recorded, and the therapeutic efficacy was evaluated after treatment.
RESULTS:
After 1, 2, 3 months of treatment, the TCM syndrome scores and YGTSS scores of the two groups were decreased compared with those before treatment (P<0.01, P<0.05), and the scores of the observation group were lower than those in the control group (P<0.05, P<0.01). The disappearance time of simple vocal tics in the observation group was earlier than that in the control group (P<0.05). The effective rate of the observation group was 93.1% (27/29), which was higher than 73.3% (22/30) in the control group (P<0.05).
CONCLUSION
Acupuncture at "four pharyngeal points" could improve symptoms in children with simple vocal tics of liver hyperactivity disturbed wind, and shorten the disappearance time of simple vocal tics.
Humans
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Male
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Acupuncture Points
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Female
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Child
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Acupuncture Therapy
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Drugs, Chinese Herbal/administration & dosage*
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Child, Preschool
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Liver/drug effects*
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Tics/drug therapy*
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Treatment Outcome
8.Erratum: Author correction to "The upregulated intestinal folate transporters direct the uptake of ligand-modified nanoparticles for enhanced oral insulin delivery" Acta Pharm Sin B 12 (2022) 1460-1472.
Jingyi LI ; Yaqi ZHANG ; Miaorong YU ; Aohua WANG ; Yu QIU ; Weiwei FAN ; Lars HOVGAARD ; Mingshi YANG ; Yiming LI ; Rui WANG ; Xiuying LI ; Yong GAN
Acta Pharmaceutica Sinica B 2025;15(6):3353-3353
[This corrects the article DOI: 10.1016/j.apsb.2021.07.024.].
9.Preparation and characterization of an injectable enamel matrix protein hydrogel
Chunbao WANG ; Ziyu ZENG ; Yaqi QUAN ; Zhongshan WANG
Journal of Practical Stomatology 2024;40(5):614-618
Objective:To prepare an enamel matrix protein(EMP)loaded poloxamer thermosensitive injectable hydrogel,and to study its characteristics.Methods:EMP was extracted from a 6-month old porcine and identified by SDS-PAGE electrophoresis.EMP at 2 mg/mL was loaded into gels poloxamer with P407∶P188 of 23∶8 and 19∶1 respectively.The gelling temperature of blank gel and EMP gel was detected by an inverted tube method.The release performance of selected EMP-loaded gel was detected by an Ultra-micro spectrophotometer.Results:A hydrogel(code number:FP1)with formula of 2 mg/mL EMP and 23∶8 of P407∶P188 possessed gel-ling temperature of 33 ℃,and the release behavior of FP1 conformed to the Higuchi equation.Conclusion:A poloxamer thermosensitive injectable hydrogel was successfully fabricated,this formulation possesses adaptive phase inversion temperature and controlled release ef-ficiency.
10.Application of customized dental-bone supported osteotomy guide plate in bilateral sagittal split osteotomy
Lidong WANG ; Wen MA ; Shuai FU ; Changbin ZHANG ; Qingying CUI ; Canbang PENG ; Yaqi CHEN ; Ming LI
Journal of Practical Stomatology 2024;40(5):698-702
Objective:To investigate the effects of customized dental-bone supported osteotomy guide plate in bilateral sagittal split osteotomy.Methods:24 patients(48 sides)with maxillofacial deformity underwent BSSO were included.The maxillofacial region of all patients was scanned by CT,the plaster dental models were scanned using laser surface scanner,and the 3D models were established.The osteotomy guide plates of the inner horizontal and anterior sagittal bone incision of mandible ramus were manufactured by digital technology.All splits underwent operation with(27 side)and without(21 sides)the osteotomy guide plate respectively by the same doc-tor,and the time for the inner horizontal and anterior sagittal bone incision of mandible ramus was recorded.Postoperative CT scan was performed to evaluate the surgical effects according to the lingual split scale(LSS)classification.Results:The wound in all patients healed well and no serious complication was observed.The time for the inner horizontal and anterior sagittal bone incision of mandible ramus in plate group and no plate group was(125.67±2.23)s and(141.15±3.69)s respectively(P<0.05).The probability of mandi-ble splitting according to Hunsuck standard osteotomy line increased from 42.86%to 66.67%,and the probability of osteotomy line passing through mandibular nerve canal decreased from 33.33%to 7.41%,by using osteotomy guide plate.In addition,LSS4 type of osteotomy line was avoided by using osteotomy guide plates.Data analysis showed that the split pattern of sagittal split osteotomy of the mandibular ramus was influenced by the application of osteotomy guide plates(P<0.05).Conclusion:The customized dental-bone supported osteotomy guide plate is effective in the completion of the operation and reducing time consuming and surgical complication in BSSO.


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