1.Research progress on digital technology in endodontic microsurgery
XU Siying ; DI Jingyi ; QU Jiongwei ; HE Li
Journal of Prevention and Treatment for Stomatological Diseases 2026;34(1):96-104
Endodontic microsurgery (EMS) is an oral surgical procedure that utilizes the magnification and illumination provided by a microscope. Fine instruments are used to remove periapical diseased tissues, resect the root apex, and tightly seal the root canal system, aiming to promote the healing of periapical tissues and retain the affected tooth whenever possible. Precise localization and resection of the root apex have always been challenging in EMS. The application of digital technology in EMS can address many issues in traditional endodontic microsurgery. Digital technology offers advantages such as optimizing surgical planning, providing precise positioning guidance, and enhancing operational accuracy. Currently, the commonly used digital technologies in EMS include static and dynamic navigation technologies and oral surgical robots. Static navigation technology enhances surgical predictability through precise preoperative planning and guided fabrication, yet is constrained by its inability to adjust during surgery. Dynamic navigation technology excels in real-time tracking and intraoperative flexibility but demands high hand-eye coordination from surgeons and may be hindered by bulky handheld devices. Oral surgical robots reduce hand tremors and human error in surgery with their high precision, stability, and ability to adjust in real time, but their clinical applicability is limited and their cost is high. In clinical practice, tailored combinations of these technologies can be applied based on case complexity. For simple cases with well-defined anatomy, static navigation alone may suffice. For complex anatomical cases, static navigation can provide initial planning, supplemented by dynamic navigation for real-time guidance or robotic systems for high-precision execution. This paper discusses the workflow, clinical application status and advantages and limitations of these three digital technologies in EMS. The continuous development of digital technology is expected to simplify the operation process, improve the navigation accuracy, and reduce the operation cost. It is believed that with continuous improvement and optimization, these technologies will effectively break through the current bottleneck of the cost of equipment, operation complexity, and accuracy enhancement. These technologies are also expected to further expand the application boundaries, providing more minimally invasive, precise, and time-saving personalized treatment solutions for endodontic diseases.
2.Network analysis of basic psychological needs and psychological behavioral problems among junior and senior high school students in Taizhou City
LIN Nan, LI Li, FU Chaowei, LIN Haijiang, YANG Yuting, LIU Yixuan, WANG Tingting, WANG Jingyi
Chinese Journal of School Health 2026;47(3):388-393
Objective:
To explore the network structure of middle school students basic psychological needs and psychological behavioral problems, and identify the core nodes within the network, as well as examine demographic subgroup differences, so as to provide support for targeted mental health interventions for adolescents.
Methods:
In September and October of 2023, a total of 2 000 junior and senior high school students were selected with multistage cluster random sampling from 8 schools in Jiaojiang District and Tiantai County, Taizhou City. An online self administered questionnaire was used to assess emotional and behavioral problems, perceived autonomy, self awareness, loneliness, and social support. The instruments included the Strengths and Difficulties Questionnaire (SDQ), Perceived Choice and Awareness of Self Scale (PCASS), Mental Health Literacy Questionnaire (MHLQ), University of California,Los Angeles Loneliness Scale (UCLA-LS), and the Multidimensional Scale of Perceived Social Support (MSPSS). A network analysis approach was employed to construct a network representing adolescents basic psychological needs and psychological behavioral problems, focusing on centrality measures and demographic subgroup differences.
Results:
A total of 418 students (20.9%) reported abnormal emotional and behavioral problems. Perceived autonomy and competence were negatively correlated with emotional problems (weights: 0.12, 0.14) and hyperactivity (weights: 0.10, 0.16). Social support showed negative correlation with peer relationship issues, hyperactivity, and conduct problems (weights: 0.16, 0.13, 0.10). Loneliness was positively correlated with emotional symptoms and peer relationship problems (weights: 0.28, 0.18). In the overall network, perceived relationships (social support and loneliness), emotional symptoms, and hyperactivity emerged as central nodes. Significant differences in network structure were observed between gender subgroups ( P =0.02). Girls internalizing issues were more influenced by loneliness and perceived autonomy frustration, while social support exhibited higher centrality in boys.
Conclusions
Perceived relationships, emotional problems, and hyperactivity are key nodes in the network of adolescents basic psychological needs and psychological behavioral problems. Loneliness demonstrates a prominent influence within the network, and the overall network exhibits gender differences.
3.Tongnao Decoction Promotes Angiogenesis and Alleviates Cerebral Ischemic Injury via PI3K/Akt/GSK-3β Signaling Pathway
Yan LIU ; Yang WU ; Wanhui PENG ; Jingyi CHEN ; Jiale GAN ; Li LI ; Yangjingyi XIA ; Yunze LI ; Zhaoyao CHEN ; Wenlei LI ; Minghua WU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(9):100-110
ObjectiveTo investigate the mechanisms of Tongnao decoction (TND) in mice with acute ischemic stroke (AIS). MethodsFifty male C57BL/6J mice were randomly divided into a sham operation group, model group, TND low-dose group (1.86 g·kg-1), TND high-dose group (3.72 g·kg-1), and butylphthalide (NBP) group (10 mg·kg-1), with 10 mice in each group. A mouse model of cerebral ischemic injury was established using photochemical thrombosis (PT). The sham operation group and model group were administered an equal volume of normal saline by gavage. All five groups were treated once daily for 14 consecutive days. Behavioral tests were performed before modeling and at the end of administration. T2-weighted imaging (T2WI) was performed 3 days after modeling to evaluate the extent of injury. Hematoxylin-eosin (HE) staining was used to observe histological changes in the cerebral cortex, and Nissl staining was used to observe neuronal morphology. Cerebral blood flow in mice was detected using a laser speckle contrast imaging (LSCI) system. Immunofluorescence staining was used to detect the cell proliferation marker bromodeoxyuridine (BrdU) and the highly glycosylated type I transmembrane glycoprotein CD34. Western blot analysis was used to detect the expression levels of phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt), glycogen synthase kinase-3β (GSK-3β), and their phosphorylation levels, as well as tight junction-related proteins zonula occludens-1 (ZO-1), Occludin, and Claudin-5 in the peri-infarct tissue. Thirty-five zebrafish were randomly divided into normal control group, model group, TND low and high dose groups (0.16, 0.32 g·L-1) and NBP group (10 μmol·L-1), with 7 in each group. A stereoscopic fluorescence microscope was used to observe vascular growth in zebrafish. ResultsImaging showed that PT caused ischemia in the right cortical region. Behavioral tests indicated that, compared with the model group, the drug-treated groups reduced the error rate of irregular balance ladder climbing on the affected side and shortened the tape removal time (P<0.05). HE staining and Nissl staining showed that, compared with the model group, the drug-treated groups exhibited reduced brain tissue damage, fewer scars, and improved neuronal morphology. LSCI results showed that the drug-treated groups partially restored cerebral blood perfusion and promoted the establishment of collateral circulation compared with the model group. Immunofluorescence staining indicated that the drug-treated groups increased the positive rates of BrdU and CD34 compared with the model group (P<0.01), promoting angiogenesis. Meanwhile, compared with the model group, the drug-treated groups upregulated the expression levels of p-PI3K, p-Akt, p-GSK-3β, and tight junction proteins ZO-1, Occludin, and Claudin-5 (P<0.05,P<0.01), and increased the number of intersegmental vessels in zebrafish (P<0.05,P<0.01). ConclusionTND can promote angiogenesis around the infarct in PT model mice by regulating the PI3K/Akt/GSK-3β signaling pathway, thereby improving cerebral ischemic injury.
4.Comparative analysis of diagnostic criteria for occupational radiation diseases between China and the International Labour Organization
Ru XUE ; Jingyi LI ; Jinxin JU ; Erdong CHEN
Journal of Environmental and Occupational Medicine 2026;43(5):651-655
The diagnosis of occupational radiation diseases, with the diagnostic criteria as main technical basis, has always been one of the emphases of China's occupational health work. This study compared the diagnostic standards for occupational radiation diseases between China and the International Labour Organization (ILO) across three aspects—types, classifications/grades, and diagnostic dose requirements—categorizing the diseases into tissue reactions and stochastic effects. ① For tissue reactions, 8 types are included both in China and ILO standards, 13 unique to China's standards, and 4 unique to the ILO standards; significant differences exist in classifaton/grading; China's standards are either consistent with or stricter (i.e., requiring higher diagnostic dose values) than the ILO standards. ② For stochastic effects (radiation-induced cancer), 12 types were included in both standards; China's standards provide more comprehensive coverage for certain malignancies (e.g., bone cancer); while 5 types were unique to the ILO standards have, some (e.g. salivary gland cancer) are primarily observed in non-occupational populations. Overall, China’s standards include more disease types and maintain more stringent diagnostic dose requirements. However, specific discrepancies require further deliberation during future revisions. Finally, based on these results, recommendations were provided for basic research and the evaluation of Chinese diagnostic criteria.
5.Comparative analysis of diagnostic criteria for occupational radiation diseases between China and the International Labour Organization
Ru XUE ; Jingyi LI ; Jinxin JU ; Erdong CHEN
Journal of Environmental and Occupational Medicine 2026;43(5):651-655
The diagnosis of occupational radiation diseases, with the diagnostic criteria as main technical basis, has always been one of the emphases of China's occupational health work. This study compared the diagnostic standards for occupational radiation diseases between China and the International Labour Organization (ILO) across three aspects—types, classifications/grades, and diagnostic dose requirements—categorizing the diseases into tissue reactions and stochastic effects. ① For tissue reactions, 8 types are included both in China and ILO standards, 13 unique to China's standards, and 4 unique to the ILO standards; significant differences exist in classifaton/grading; China's standards are either consistent with or stricter (i.e., requiring higher diagnostic dose values) than the ILO standards. ② For stochastic effects (radiation-induced cancer), 12 types were included in both standards; China's standards provide more comprehensive coverage for certain malignancies (e.g., bone cancer); while 5 types were unique to the ILO standards have, some (e.g. salivary gland cancer) are primarily observed in non-occupational populations. Overall, China’s standards include more disease types and maintain more stringent diagnostic dose requirements. However, specific discrepancies require further deliberation during future revisions. Finally, based on these results, recommendations were provided for basic research and the evaluation of Chinese diagnostic criteria.
6.Performance Evaluation and Resource Utilization Optimization of Multidisciplinary Team Model for Lung Cancer: A Real-World Study
Meng WANG ; Xiaoli ZHANG ; Jue LIU ; Jingyi TANG ; Ziming LI
Medical Journal of Peking Union Medical College Hospital 2026;17(3):637-645
To compare the performance differences between the multidisciplinary team (MDT) model and the conventional diagnostic and treatment model for lung cancer, and to explore a high-quality development pathway for optimizing lung cancer diagnostic and treatment resources. A retrospective analysis was conducted on electronic medical record data of lung cancer patients at Shanghai Chest Hospital from March 2025 to December 2025. Patients were divided into an MDT group and a conventional care group based on whether they were admitted to the integrated oncology ward. Statistical analyses were performed using the Mann-Whitney A total of 4, 758 patients with primary lung cancer were included, comprising 365 (7.7%) in the MDT group and 4, 393 (92.3%) in the conventional care group. After adjusting for confounding factors, the MDT model significantly reduced hospitalization frequency during the observation period by 48.8% ( The MDT model for lung cancer significantly reduces hospitalization frequency; however, its effect on cost per hospitalization is population-selective, with increased costs in early-stage (stage Ⅰ) patients and decreased costs in late-stage (stages Ⅱ and Ⅳ) patients. The implementation of the MDT model should adopt precise patient stratification management, prioritizing the optimal patient population to achieve the optimal allocation of medical resources.
7.Acupuncture Treatment for Severe Bell's Palsy and Its Impact on Serum GDNF and NGF:A Randomized Controlled Trial
Li MA ; Xiaonan LI ; Chenyang SU ; Juanjuan FENG ; Jingyi LIU ; Haoyi QIAO ; Peng BAI
Journal of Traditional Chinese Medicine 2026;67(12):1297-1304
ObjectiveTo evaluate the clinical efficacy and safety of acupuncture in treating severe Bell's palsy and to explore its potential mechanism by investigating the effect on serum levels of glial cell line-derived neurotrophic factor (GDNF) and nerve growth factor (NGF). MethodsA randomized, subject-blinded, sham-acupuncture controlled trial was conducted. A total of 130 patients with severe Bell's palsy were randomly allocated into a treatment group or a control group at a 1∶1 ratio. Both groups received conventional western medicine. In addition, the treatment group received acupuncture, while the control group received sham acupuncture, with each session lasting 30 minutes. The treatment course lasted 8 weeks for both groups, followed by a follow-up assessment at week 12. The primary outcome was the proportion of patients achieving House-Brackmann (H-B) grade Ⅱ or lower at week 8. Secondary outcomes included Sunnybrook facial grading system scores at week 0, 4, 8, and 12, the time to satisfactory recovery(the time required to achieve H-B grade≤Ⅱ), distribution of H-B grades and facial disability index (FDI) scores including the physical function subscale (FDIP) and social/well-being function subscale (FDIS) scores at week 0, 4, 8, and 12, and serum GDNF and NGF levels at week 0, 4, and 8. Adverse events and participants' self-assessments of treatment efficacy were also recorded. ResultsA total of 122 participants completed the study, including 62 in the treatment group and 60 in the control group. An intention-to-treat (ITT) analysis was performed, and missing data were handled using the last observation carried forward (LOCF) method. The proportion of patients achieving H-B grade ≤grade Ⅱ at week 8 was 78.5% (51/65) in the treatment group, significantly higher than 49.2% (32/65) in the control group (P<0.05). The Sunnybrook scores, FDIP and FDIS scores increased, while H-B grades decreased at week 4, 8, and 12 in both groups compared to week 0; moreover, improvements in all outcome measures were significantly greater in the treatment group than in the control group (P<0.05). The median time to satisfactory recovery was 6 weeks (95%CI: 5.697-6.303) in the treatment group, significantly shorter than 12 weeks (95%CI: 8.314-15.686) in the control group (P<0.05). Serum levels of GDNF and NGF were significantly higher in the treatment group at weeks 4 and 8 (P<0.05). No serious acupuncture-related adverse events occurred in either group. Adverse events were reported in 5 patients (7.69%) in the treatment group and 4 patients (6.15%) in the control group, with no statistically significant difference between groups (P>0.05). Patients' self-assessment of treatment efficacy after 8 weeks treatment was significantly better in the treatment group (P<0.05). ConclusionAcupuncture can effectively improve facial nerve function and shorten recovery time in patients with severe Bell's palsy, with a favorable safety profile. The therapeutic mechanism may be associated with the upregulation of serum GDNF and NGF levels.
8.Qiangjing Tablets Regulate CDK4-E2F Signaling Pathway to Delay Aging of Leydig Cells and Testicular Tissue in Rats
Xiucheng LAN ; Meijing WANG ; Jingyi ZHANG ; Junjun LI ; Liang DONG ; Xujun YU ; Fang YANG ; Degui CHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(14):328-336
ObjectiveTo reveal the molecular mechanism by which the traditional Chinese medicine compound prescription Qiangjing tablets regulate the aging of the testicular tissue and Leydig cells in rats through the cyclin-dependent kinase 4 (CDK4)-early 2 factor (E2F) signaling pathway. MethodsFor the cell experiment, 2-month-old SPF-grade SD male rats were selected and randomly assigned into a blank control group (administrated with an equal volume of 0.9% sodium chloride injection) and a Qiangjing tablets group (20 rats in each group) according to body weight. The Leydig cell model of aging was established by treatment of TM3 cells with 100 μmol·L-1 H2O2, and the modeling performance was evaluated based on the levels of p16 and p21 determined by Western blot. The antioxidant NAC (1 mmol·L-1) was used as the positive control for eliminating reactive oxygen species (ROS). Cells were intervened with Qiangjing tablets-containing serum at low (2.5%), medium (5%), and high (10%) concentrations. The testosterone level in the cell supernatant was determined by enzyme-linked immunosorbent assay (ELISA), and the protein levels of CDK4, E2F1, and E2F2 were analyzed by Western blot. In the animal experiment, 19-month-old naturally aging rats were used as the model group, and 2-month-old rats as the young control group. The positive control group was subcutaneously injected with 5.21 mg·kg-1·d-1 testosterone propionate. Qiangjing tablets were administered by gavage at low, medium, and high doses of 0.72, 1.44, 2.88 g·kg-1·d-1, respectively. The general conditions of rats were observed, and the protein levels of CDK4, E2F1, and E2F2 in the testicular tissue were determined by Western blot. ResultsIn the cell experiment, compared with the blank control group, the model group showed upregulated expression of CDK4 and E2F1 (P<0.05) and slightly downregulated expression of E2F2. Compared with that in the model group, the expression of CDK4 was upregulated in the NAC group and the low-dose Qiangjing tablets group (P<0.05), slightly upregulated in the medium-dose Qiangjing tablets group, and downregulated in the high-dose Qiangjing tablets group (P<0.05). The NAC group showed downregulated expression of E2F1 (P<0.05) and E2F2, and the low-, medium-, and high-dose Qiangjing tablets groups showed downregulated expression of both E2F1 and E2F2 (P<0.05). Compared with that in the NAC group, the expression of CDK4 was upregulated in the low-dose Qiangjing tablets group and downregulated in the medium-dose and high dose (P<0.05) groups. The expression of E2F1 was down-regulated in all the three dose groups, with statistically significance in the high dose group (P<0.05), and that of E2F2 were downregulated in all the three dose groups (P<0.05). In the animal experiment, compared with the young control group, the model group exhibited downregulated expression of CDK4 (P<0.05) and slightly upregulated expression of E2F1 and E2F2. Compared with that in the model group, the expression of CDK4 decreased in the testosterone propionate group and the low-dose Qiangjing tablets group (P<0.05) but increased in the medium-dose (P<0.05) and high-dose groups. In addition, the expression of E2F1 decreased (P<0.05), and that of E2F2 was slightly elevated. Compared with that in the NAC group, CDK4 expression was elevated in the Qiangjing tablets groups, with statistical significance in the medium- and high-dose groups (P<0.05). Similarly, the E2F1 expression was also upregulated in the Qiangjing tablets groups, with statistical significance in the medium-dose group (P<0.05). The expression of E2F2 was downregulated in all the Qiangjing tablets groups. ConclusionQiangjing tablets delay the aging process of Leydig cells and testicular tissue by up-regulating the expression of CDK4 and lowering the levels of E2F1 and E2F2.
9.Qiangjing Tablets Alleviate Oxidative Stress Damage in Varicocele by Regulating Keap1/Nrf2 Signaling Pathway
Liang DONG ; Fang YANG ; Jingyi ZHANG ; Xinyi TANG ; Yulin LI ; Xujun YU ; Degui CHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(14):347-359
ObjectiveTo explore the mechanism by which Qiangjing tablets (QJT) alleviate the spermatogenic function damage caused by varicocele (VC) based on the Kelch-like ECH-associated protein 1 (Keap1)/nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway-mediated oxidative stress. MethodsTen Sprague-Dawley (SD) rats were randomly assigned into a control group and a model group. Pathological examination confirmed the stability of the model. Thirty-six SD rats were randomized into control, model, low-dose (0.23 g·kg-1) QJT, medium-dose (0.46 g·kg-1) QJT, high-dose (0.92 g·kg-1) QJT, and mazhilin (61.7 mg·kg-1) groups, with 6 rats in each group. A rat model of experimental left varicocele (ELV) was established by partially ligating the left renal vein to simulate the human nutcracker syndrome. The rats were administrated with corresponding agents once a day for 28 consecutive days. The in vitro testicular culture model of rats was established through the Transwell chamber method and intervened with QJT-containing sera (2.3, 4.6, and 9.2 g·kg-1). Microscopic observation was carried out for the morphology of the left kidney. A micrometer was used to measure the diameter of the left spermatic vein (LSV). The body weights of rats were recorded weekly, and the epididymis and testis weights were measured. The pathological changes of the testicular tissue was observed via hematoxylin-eosin (HE) staining. The levels of testosterone (T) in the cell culture supernatant and reactive oxygen species (ROS) in the rat testicular tissue were measured by enzyme-linked immunosorbent assay (ELISA). Flow cytometry was employed to determine the ROS content. Immunohistochemical staining was conducted to analyze Keap1, Nrf2, 3β-hydroxysteroid dehydrogenase (3β-Hsd), GATA-binding protein-4 (Gata-4), and proto-oncogene receptor tyrosine kinase (C-kit). The ultrastructure of the tissue was observed by transmission electron microscopy (TEM). Cell apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining. The expression of Keap1, Nrf2, glutathione S-transferase α2 (Gsta2), glutathione S-transferase μ1 (Gstm1), heme oxygenase-1 (HO-1), quinone oxidoreductase 1 (Nqo1), and thioredoxin reductase 1 (Txnrd1) was quantified by Real-time quantitative polymerase chain reaction(Real-time PCR) and Western blot. ResultsCompared with the control group, the ROS content and the percentage of apoptotic cells in the model group were significantly increased (P<0.01), the T concentration was significantly decreased (P<0.01), the mRNA and protein expressions of Keap1 were significantly increased (P<0.01), and the mRNA and protein expressions of Nrf2, Gsta2, Gstm1, HO-1, Nqo1 and Txnrd1 were significantly decreased (P<0.05). Compared with the model group, the ROS content and the percentage of apoptotic cells in each dose group of the Qiangjing Tablets were significantly reduced (P<0.05), and the mRNA and protein expressions of Keap1 were significantly decreased (P<0.05), while the mRNA and protein expressions of Nrf2, Gsta2, Gstm1, HO-1, Nqo1 and Txnrd1 were significantly increased (P<0.05). ConclusionQJT improves sperm motility in the rat model of VC by modulating the Keap1/Nrf2 signaling pathway and reducing oxidative stress injury.
10.Mechanism of Quercetin-loaded Exosomes in Improving Testosterone Synthesis in Leydig Cells from Correlation Perspective of "Disease, Syndrome, Formula, and Medicine"
Meijing WANG ; Xiucheng LAN ; Fangyue WANG ; Jingyi ZHANG ; Guangsen LI ; Degui CHANG ; Xujun YU ; Fang YANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(14):360-370
ObjectiveBased on the multidimensional correlation analysis framework of "disease, syndrome, formula, and medicine", this study aims to systematically elucidate the regulatory effects of effective components in Qiangjing tablet on testosterone synthesis pathways in testicular Leydig cells under oxidative stress, providing a theoretical basis for the treatment of male infertility with traditional Chinese medicine and modern research on compounds. MethodsDisease targets for male infertility were obtained from The Human Gene Database (GeneCards, score ≥20), the Comparative Toxicogenomics Database (CTD, score ≥150), DrugBank (score ≥0.2), and DisGeNET (score ≥0.2). Targets related to the syndrome of kidney deficiency and blood stasis were acquired from the traditional Chinese medicine syndrome association database SymMap. Components of Qiangjing tablet were retrieved based on The Encyclopedia of Traditional Chinese Medicine (ETCM) database and the Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine (TCMIP), and they were screened according to a quantitative estimate of drug-likeness (QED ≥ 0.49) and a target confidence index>0.8. Intersecting targets were taken to construct a protein-protein interaction (PPI) network using the STRING database. The network was visualized with Cytoscape software and subjected to the functional annotation of gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis. Quality markers (Q-markers) were predicted via the ADMETlab 2.0 platform based on Lipinski's rule, Pfizer's rule, GSK's rule, and the Golden Triangle. For experimental validation, rats' testicular Leydig cells were used. Exosomes were extracted and loaded with active components via the ultrasonic method. Exosome concentration was determined using a BCA protein quantification kit. Morphology was observed using a transmission electron microscope. The particle size was analyzed with a particle size analyzer. The surface marker proteins such as cluster of differentiation 9 (CD9), cluster of differentiation 63 (CD63), and cluster of differentiation 81 (CD81) were identified by Western blot, and drug loading capacity was measured by high-performance liquid chromatography (HPLC). An oxidative stress model was induced by alpha, alpha'-azodiisobutyramidine dihydrochloride (AAPH), and Leydig cells were divided into the following groups: A control group, an AAPH group, a quercetin group (Que group), an exosome group (Exo group), and a QUE-loaded Exo group (Que-Exo group). The cell viability was detected using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) thiazolyl blue assay. The reactive oxygen species (ROS) levels and mitochondrial membrane potential were measured by flow cytometry. The levels of oxidative indicators, including malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT), and testosterone (T), were detected by enzyme-linked immunosorbent assay (ELISA). The expressions of steroidogenic enzymes such as cytochrome p450 family 11 subfamily a member 1 (CYP11A1), hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid delta-isomerase 1 (HSD3B1), and hydroxysteroid 17-beta dehydrogenase 3 (HSD17B3), regulatory factors such as steroidogenic factor 1 (SF-1) and steroidogenic acute regulatory protein (StAR), and miR-145-5p content, were detected by Western blot and real-time polymerse chain reaction (Real-time PCR). ResultsNetwork pharmacology analysis reveals that the main active components of Qiangjing tablet for intervening in male infertility with kidney deficiency and blood stasis syndrome were Que, luteolin, etc., with the core mechanism involving pathways such as steroid hormone biosynthesis. Experimental results show that compared with the control group, the AAPH group exhibits significantly reduced cell viability (P<0.01), decreased mitochondrial membrane potential (P<0.01), significantly elevated levels of ROS, MDA, and miR-145-5p (P<0.01), significantly reduced activities of SOD, GSH-Px, and CAT, as well as reduced testosterone content (P<0.01), and significantly downregulated protein and mRNA expressions of steroidogenic enzymes, SF-1, and StAR (P<0.01). The above indicators were reversed in the Que and Que-Exo groups (P<0.05). Compared with the Que group, the Que-Exo group showed more significant effects in enhancing cell viability, mitochondrial membrane potential, testosterone level, antioxidant enzyme activities, and expressions of key molecules in the steroidogenic pathway (P<0.05). ConclusionThis study demonstrates that Que, an active component of Qiangjing tablet, inhibits oxidative stress reaction, improves mitochondrial function in Leydig cells, upregulates steroidogenic enzyme expression, and restores testosterone production. As a carrier for Que, Exo enhance its stability, delivery efficiency, and biological effect. Additionally, miR-145-5p may be closely associated with testosterone synthesis, though its precise molecular mechanism requires further exploration. By integrating traditional Chinese medicine compounds with modern scientific technology, this research expands the paths for the modernized research of traditional Chinese medicine and opens a novel therapeutic direction with translational potential for clinical intervention of male infertility.


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