1.Therapeutic effect of neurosurgical robot-assisted stereotactic puncture and drainage for brain abscess
Xu RAN ; Jing-peng LIU ; Ju-hong PENG ; Zuo-xin ZHANG ; Yuan XIE ; Yan XIANG ; Lin YANG ; Jin-bo YIN ; Guo-long LIU ; Sheng-qing LYU
Journal of Regional Anatomy and Operative Surgery 2025;34(11):987-992
Objective To evaluate the clinical outcome of neurosurgical robot-assisted stereotactic puncture and drainage for brain abscess.Methods A retrospective analysis was conducted on the clinical data of 53 patients with brain abscess admitted to our hospital from January 2018 to December 2024.Among them,29 cases underwent craniotomy for abscess resection(craniotomy group),while 24 cases received neurosurgical robot-assisted stereotactic puncture and drainage(robot-assisted group).The operation time,intraoperative blood loss,decompressive craniectomy rate,proportion of postoperative antibiotic regimen adjustment,postoperative hospital stay,incidence of postoperative complications,mortality rate and Glasgow outcome scale(GOS)scores 6 months after surgery of patients were compared between the two groups.Results Compared with the craniotomy group,the robot-assisted group demonstrated significantly shorter operation time,less intraoperative blood loss,and lower incidence of postoperative complication,the differences were all statistically significant(P<0.05).However,there were no statistically significant differences in terms of decompressive craniectomy rate,postoperative hospital stay,mortality rate,GOS score,or proportion of the postoperative antibiotic regimen adjustment between the two groups(P>0.05).Conclusion As a precise and minimally invasive surgical method,neurosurgical robot-assisted stereotactic puncture and drainage for patients with brain abscess can effectively improve the operational efficiency,shorten the operation time,reduce intraoperative injury,and lower the risk of postoperative complications.It has high clinical application value and potential for widespread adoption.
2.Construction and evaluation of a cell model simulating the change of testicular microenvironment mediated by hypoxic and high-pressure conditions in varicocele mice
Shu-lin LIANG ; Li-guo GENG ; Ling HAN ; Chu-nan RONG ; Zhan QIN ; Juan DU ; Chao-ba HE ; Shao-ying YUAN
National Journal of Andrology 2025;31(6):483-491
Objective:Varicocele(VC)induces male infertility by mediating changes in the testicular microenvironment,in which testicular hypoxia and high-pressure are important pathological conditions.This study aims to compare the mouse spermatogenesis(GC-2spd)cells and Sertoli(TM4)cells of mouse testis after hypoxic modeling and hypoxic and high-pressure combined modeling,and to explore the feasibility of establishing a hypoxic and high-pressure combined cell model.Methods:On the basis of cell hypoxia induced by CoCl2,the complex model of testicular cell hypoxia and high pressure was constructed by changing the osmotic pressure of GC-2 and TM4 cell medium with a high concentration of NaCl solution.After selecting the intervention concentration of CoCl2 by MTT test and detecting the expression level of HIF-1α for the determination of the optimal osmotic pressure conditions of the cell model,the cells were divided into normal group,hypoxia model group and composite model group.And the levels of OS,programmed cell death,inflammatory factors,and the expression levels of pyroptosis-related proteins were compared between the normal group and the groups with different modeling methods.Results:The optimal intervention concentration of CoCl2 in GC-2 and TM4 cells was 150 and 250μmol/L,respectively,and the expression of HIF-1α was the highest in both cells under osmotic pressure of 500 mOsmol/kg(P<0.05).Compared with the normal group,the SOD levels of GC-2 and TM4 cells decreased(all P<0.05),CAT level decreased(all P<0.05),and MDA level increased(all P<0.01),and the OS level of GC-2 and TM4 cells was more obvious than that of the hy-poxia model group(all P<0.05).Compared with the normal group,apoptosis occurred in GC-2 and TM4 cells after composite model-ing(all P<0.05).Compared with the normal group,the mRNA expressions of IL-1β,IL-18,TNF-α and COX-2 in GC-2 and TM4 cells significantly increased(P<0.01)and higher than those in hypoxia model group(P<0.05)and induced pyroptosis(P<0.01).The expression level of GSDMD increased(P<0.05).Conclusion:The cell model with hypoxia and high pressure com-bined modeling can not only induce oxidative stress and apoptosis of cells better than that with hypoxia alone,but also further cause in-flammatory response damage and pyroptosis,which simulates the changes of testis microenvironment mediated by hypoxia and high pressure combined conditions in VC.This cell model can be used for studying the pathogenesis of VC-associated male infertility,evalu-ating drug efficacy,and exploring pharmacological mechanisms.
3.Therapeutic effect of neurosurgical robot-assisted stereotactic puncture and drainage for brain abscess
Xu RAN ; Jing-peng LIU ; Ju-hong PENG ; Zuo-xin ZHANG ; Yuan XIE ; Yan XIANG ; Lin YANG ; Jin-bo YIN ; Guo-long LIU ; Sheng-qing LYU
Journal of Regional Anatomy and Operative Surgery 2025;34(11):987-992
Objective To evaluate the clinical outcome of neurosurgical robot-assisted stereotactic puncture and drainage for brain abscess.Methods A retrospective analysis was conducted on the clinical data of 53 patients with brain abscess admitted to our hospital from January 2018 to December 2024.Among them,29 cases underwent craniotomy for abscess resection(craniotomy group),while 24 cases received neurosurgical robot-assisted stereotactic puncture and drainage(robot-assisted group).The operation time,intraoperative blood loss,decompressive craniectomy rate,proportion of postoperative antibiotic regimen adjustment,postoperative hospital stay,incidence of postoperative complications,mortality rate and Glasgow outcome scale(GOS)scores 6 months after surgery of patients were compared between the two groups.Results Compared with the craniotomy group,the robot-assisted group demonstrated significantly shorter operation time,less intraoperative blood loss,and lower incidence of postoperative complication,the differences were all statistically significant(P<0.05).However,there were no statistically significant differences in terms of decompressive craniectomy rate,postoperative hospital stay,mortality rate,GOS score,or proportion of the postoperative antibiotic regimen adjustment between the two groups(P>0.05).Conclusion As a precise and minimally invasive surgical method,neurosurgical robot-assisted stereotactic puncture and drainage for patients with brain abscess can effectively improve the operational efficiency,shorten the operation time,reduce intraoperative injury,and lower the risk of postoperative complications.It has high clinical application value and potential for widespread adoption.
4.Clinicopathological analysis of ALK-positive histiocytosis involving central nervous system
Yi-fei REN ; Mei-lin ZHANG ; Yuan-yuan CHENG ; Ji XIONG ; Yin WANG ; Feng TANG ; Zun-guo DU
Fudan University Journal of Medical Sciences 2025;52(6):837-846
Objective To summarize and analyze the clinicopathological characteristics,imaging manifestations,treatment and prognosis of anaplastic lymphoma kinase(ALK)-positive histiocytosis(APH)involving the central nervous system(CNS),so as to enhance understanding of this rare disease.Methods A retrospective analysis was conducted on 5 cases of CNS-involved APH diagnosed in Huashan Hospital,Fudan University between 2019 and 2023.Clinical,imaging and pathological data were collected,and supplemented by immunohistochemical staining(IHC),fluorescence in situ hybridization(FISH)and high-throughput sequencing for auxiliary diagnosis and molecular characterization.The findings were summarized and integrated with previous literature for a comprehensive analysis.Results All five patients were male,with a mean age of 27.6 years,in which 2 cases exhibited multi-system involvement,while 3 cases exhibited single system involvement.Imaging revealed multiple intracranial lesions in multi-system cases and solitary lesions in single system cases,with well-defined boundaries and homogeneous enhancement.Histologically,tumor cells were intermingled with lymphocytes,displaying an alternating"light and dark"pattern in 1 case.Granuloma-like structures were observed in 4 cases,along with frequent nuclear grooves,indentations and convolutions.Tumor cells usually infiltrated surrounding tissues.IHC demonstrated that tumors expressed histiocytic markers(CD68/CD163)and ALK predominantly expressed in the cytoplasm.FISH confirmed ALK gene rearrangements in all patients,while high-throughput sequencing identified KIF5B-ALK fusions in 2 cases.All single system CNS cases achieved complete remission after surgical resection with or without adjuvant chemotherapy/ALK inhibitors.Among multi-system cases,one achieved partial remission,and one experienced relapse and progression.Conclusion CNS APH is prone to preoperative misdiagnosis.Its histopathological features,ALK immunohistochemical expression,and ALK gene fusions are critical for diagnosis.Patients with single system involvement demonstrate overall superior outcomes compared to multi-system cases.Total resection is effective for localized disease,while multi-system cases require systemic therapy.Multidisciplinary collaboration is crucial for precise diagnosis and treatment.
5.Disease Burden and Associated Risk Factors of Early-Onset Lung Cancer in China and Worldwide
Lin CAI ; Chenxin ZHU ; Jiani YUAN ; Xinglong ZHANG ; Yi FANG ; Haiyan YANG ; Lanwei GUO
Medical Journal of Peking Union Medical College Hospital 2025;16(4):1047-1056
Objective To assess the global and Chinese disease burden of early-onset lung cancer(di-agnosed in patients aged 15-49 years)and its major risk factors.Methods Based on the GLOBOCAN 2022 and Global Burden of Disease(GBD)2021 datasets,we evaluated the disease burden and associated risk fac-tors of early-onset lung cancer globally and in China,stratified by age,sex,geographic location,and human development index(HDI).Key indicators included age-standardized incidence rate(ASIR),age-standardized mortality rate(ASMR),and disability adjusted life years(DALYs)attributable to risk factors.Results In 2022,there were 137 705 new cases and 72 646 deaths from early-onset lung cancer globally,with ASIR and ASMR of 3.43 per 100 000 and 1.82 per 100 000 population,respectively.The disease burden was higher in males than in females(ASIR:3.72 per 100 000 vs.3.14 per 100 000;ASMR:2.31 per 100 000 vs.1.33 per 100 000).High-HDI regions exhibited the highest ASIR(5.51 per 100 000)and ASMR(2.57 per 100 000),with health inequality analysis revealing a concentration of disease burden in higher-HDI areas.China bore the heaviest burden,accounting for 48.69%of global new cases and 35.77%of deaths.China's ASIR(8.21 per 100 000)and ASMR(3.17 per 100 000)exceeded global averages,with incidence higher in fe-males(8.78 per 100 000 vs.7.67 per 100 000)but mortality higher in males(4.01 per 100 000 vs.2.29 per 100 000).Smoking and ambient particulate matter pollution were the leading risk factors globally(DALYs contribution:42.01%and15.62%)and in China(DALYs contribution:46.78%and 20.84%).Globally,household air pollution ranked third,whereas in China,secondhand smoke replaced it as the third leading risk factor,with household air pollution dropping to fifth.Risk factor profiles varied significantly across age groups,with modifiable risks contributing less to disease burden in the 15-24 age group.Conclusions The burden of early-onset lung cancer varies markedly by sex,region,and HDI,with China facing a disproportionately high burden.Policymakers should prioritize equitable resource allocation and targeted interventions,particularly in tobacco control and air pollution mitigation,to enhance cancer prevention and control efforts.
6.Distal derotational femoral osteotomy combined with knee extension device reconstruction for adolescent habitual patellar dislocation with severe lower limb torsional deformity
Chao FENG ; Lin HUANG ; Lianyang LIN ; Shengzhong WU ; Yukun WANG ; Yuan GUO ; Xuemin LYU ; Zheng YANG
Chinese Journal of Orthopaedics 2025;45(10):654-661
Objective:To investigate the surgical technique and initial outcomes of distal derotational femoral osteotomy (DDFO) combined with knee extension device reconstruction in adolescents with habitual patellar dislocation and severe lower limb torsional deformity.Methods:A retrospective study was conducted on 10 adolescent patients (12 knees) treated at Beijing Jishuitan Hospital and its Guizhou branch from June 2016 to June 2022. There were 6 males and 4 females with an average age of 12.0±1.5 years (range: 10.0-14.5 years) Surgical treatment included DDFO and knee extension device reconstruction (lateral retinacular release, medial retinacular plication, Roux-Goldthwait distal realignment, and MPFL reconstruction). Clinical outcomes were assessed using Lysholm scores, incidence of redislocation and complications, and imaging parameters (lateral patellofemoral angle, Insall-Salvati index, TT-TG distance, and femoral anteversion angle) preoperatively and at 1 year postoperatively.Results:All 12 knees were successfully operated on, with an average surgery time of 2.0±0.5 h (range 1.0-2.5 h), intraoperative blood loss of 47.1±17.1 ml (range 20-80 ml), and follow-up time of 46.2±18.7 months (range 24-72 months). The Lysholm knee score improved from 58.25±8.80 preoperatively to 89.17±5.32 at final follow-up ( t=-9.096, P<0.001) with significant difference. The lateral patellofemoral angle improved from -64.92±4.68 preoperatively to 6.08±2.27 at final follow-up ( t=39.178, P<0.001) with significant difference. The femoral anteversion angle decreased from 34.08±3.06 preoperatively to 14.50±2.65 at final follow-up ( t=16.916, P<0.001) with significant difference. No patellar redislocation, skin necrosis, wound infection, or limited joint mobility occurred during follow-up. Conclusion:DDFO combined with knee extension device reconstruction is an effective and safe treatment for adolescent habitual patellar dislocation with severe torsional deformity, resulting in significant clinical and radiographic improvement with low complication rates.
7.Analysis of factors influencing frequent episodes in children with moderate-to-severe atopic dermatitis: a national multicenter cross-sectional study
Jing TIAN ; Yifeng GUO ; Xiaoyan LUO ; Yuan LIANG ; Ping LI ; Jinping CHEN ; Yao LU ; Jianping TANG ; Yunsheng LIANG ; Ying GAO ; Qiufang QIAN ; Hong SHU ; Hongxiang CHEN ; Pingshen FAN ; Xiuping HAN ; Hua QIAN ; Qinfeng LI ; Ming LI ; Shengchun WANG ; Ying LIU ; Hua WANG ; Lin MA
Chinese Journal of Dermatology 2025;58(10):943-951
Objective:To investigate factors influencing frequent episodes (≥ 4 episodes within 1 year) in children with moderate-to-severe atopic dermatitis (AD) in China.Methods:A national multicenter cross-sectional study was conducted. Patients under the age of 18 years diagnosed with moderate-to-severe AD were enrolled at dermatology clinics in 18 medical institutions across 12 provinces and municipalities in China between June 12 and August 8, 2023. At the time of the visit, their guardians completed a structured questionnaire covering demographic characteristics, clinical features of AD, personal and family history, factors associated with frequent episodes of moderate-to-severe AD, compliance with treatment, and disease awareness. Statistical analyses included t tests, one-way analysis of variance, rank-sum tests, and chi-square tests, with multiple-response analysis applied for multiple-choice questions. Results:A total of 965 valid questionnaires were collected, and 965 children with moderate-to-severe AD were included. Among them, there were 531 males and 434 females, 678 (70.3%) were aged 2 - < 12 years, 837 (86.7%) were from urban areas, the age at onset was 2.47 ± 3.03 years, and the median frequency of AD episodes in the past year was 4 times. These children were divided into 2 groups based on the median episode frequency: < 4-episode group (439 cases, 45.5%) and ≥ 4-episode group (526 cases, 54.5%). Compared with the < 4-episode group, children in the ≥ 4-episode group showed younger ages at onset (2.22 ± 2.98 years vs. 2.76 ± 3.06 years, P = 0.006) and higher proportions of patients with comorbid allergic diseases in both the children themselves (82.9% [436/526] vs. 69.7% [306/439], χ2 = 23.42, P < 0.001) and their relatives (66.0% [347/526] vs. 57.4% [252/439], χ2 = 7.46, P = 0.006). Children in the ≥ 4- episode group also had higher monthly usage of moisturizers (150 [30, 300] g vs. 60 [6, 200] g) and daily frequency of moisturizer use, greater disease awareness, but more severe fear of medication use (all P < 0.05). The region and the human development index level were both significantly associated with the episode frequency (both P < 0.001), with the highest proportion of children from South China in the ≥ 4- episode group (36.3%, 191/526). Children in the ≥ 4-episode group also had a longer duration of topical glucocorticoid use than those in the < 4-episode group ( Z = -2.21, P = 0.027). External triggers associated with AD episodes mainly included heat exposure (50.36%, 486/965), hot water bathing (40.73%, 393/965), seafood (23.52%, 227/965), and dust mites (33.37%, 322/965) . Conclusion:In children with moderate-to-severe AD in China, factors influencing frequent episodes may include residence in southern or economically developed regions, earlier age at onset, having a personal or family history of allergic diseases, and fear of medication use.
8.Regulation of Immune Function by Exercise-induced Metabolic Remodeling
Hui-Guo WANG ; Gao-Yuan YANG ; Xian-Yan XIE ; Yu WANG ; Zi-Yan LI ; Lin ZHU
Progress in Biochemistry and Biophysics 2025;52(6):1574-1586
Exercise-induced metabolic remodeling is a fundamental adaptive process whereby the body reorganizes systemic and cellular metabolism to meet the dynamic energy demands posed by physical activity. Emerging evidence reveals that such remodeling not only enhances energy homeostasis but also profoundly influences immune function through complex molecular interactions involving glucose, lipid, and protein metabolism. This review presents an in-depth synthesis of recent advances, elucidating how exercise modulates immune regulation via metabolic reprogramming, highlighting key molecular mechanisms, immune-metabolic signaling axes, and the authors’ academic perspective on the integrated “exercise-metabolism-immunity” network. In the domain of glucose metabolism, regular exercise improves insulin sensitivity and reduces hyperglycemia, thereby attenuating glucose toxicity-induced immune dysfunction. It suppresses the formation of advanced glycation end-products (AGEs) and interrupts the AGEs-RAGE-inflammation positive feedback loop in innate and adaptive immune cells. Importantly, exercise-induced lactate, traditionally viewed as a metabolic byproduct, is now recognized as an active immunomodulatory molecule. At high concentrations, lactate can suppress immune function through pH-mediated effects and GPR81 receptor activation. At physiological levels, it supports regulatory T cell survival, promotes macrophage M2 polarization, and modulates gene expression via histone lactylation. Additionally, key metabolic regulators such as AMPK and mTOR coordinate immune cell energy balance and phenotype; exercise activates the AMPK-mTOR axis to favor anti-inflammatory immune cell profiles. Simultaneously, hypoxia-inducible factor-1α (HIF-1α) is transiently activated during exercise, driving glycolytic reprogramming in T cells and macrophages, and shaping the immune landscape. In lipid metabolism, exercise alleviates adipose tissue inflammation by reducing fat mass and reshaping the immune microenvironment. It promotes the polarization of adipose tissue macrophages from a pro-inflammatory M1 phenotype to an anti-inflammatory M2 phenotype. Moreover, exercise alters the secretion profile of adipokines—raising adiponectin levels while reducing leptin and resistin—thereby influencing systemic immune balance. At the circulatory level, exercise improves lipid profiles by lowering pro-inflammatory free fatty acids (particularly saturated fatty acids) and triglycerides, while enhancing high-density lipoprotein (HDL) function, which has immunoregulatory properties such as endotoxin neutralization and macrophage cholesterol efflux. Regarding protein metabolism, exercise triggers the expression of heat shock proteins (HSPs) that act as intracellular chaperones and extracellular immune signals. Exercise also promotes the secretion of myokines (e.g., IL-6, IL-15, irisin, FGF21) from skeletal muscle, which modulate immune responses, facilitate T cell and macrophage function, and support immunological memory. Furthermore, exercise reshapes amino acid metabolism, particularly of glutamine, arginine, and branched-chain amino acids (BCAAs), thereby influencing immune cell proliferation, biosynthesis, and signaling. Leucine-mTORC1 signaling plays a key role in T cell fate, while arginine metabolism governs macrophage polarization and T cell activation. In summary, this review underscores the complex, bidirectional relationship between exercise and immune function, orchestrated through metabolic remodeling. Future research should focus on causative links among specific metabolites, signaling pathways, and immune phenotypes, as well as explore the epigenetic consequences of exercise-induced metabolic shifts. This integrated perspective advances understanding of exercise as a non-pharmacological intervention for immune regulation and offers theoretical foundations for individualized exercise prescriptions in health and disease contexts.
9.Optimization and Mechanism Exploration of Tusizi Prescription for Ovarian Reserve Function Based on Uniform Design Method
Yuan LI ; Hanqian DU ; Jiashan LI ; Li GUO ; Zehui LI ; Na LIN ; Ying XU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(7):53-62
ObjectiveTo optimize Tusizi prescription for ovarian reserve function based on the uniform design method combined with in vitro experiments and explore the underlying mechanisms of this prescription. MethodsThe uniform design method was adopted to design a 5-factor 11-level experiment on the water extract of Tusizi prescription. The cell-counting kit-8 (CCK-8) assay was employed to measure the viability of human ovarian granulosa cells (KGN cells) treated with Tusizi prescription extracts 1-11, and multivariate regression analysis was performed to determine the optimal herb ratio in this prescription. The potential targets of active ingredients in the prescription were retrieved from traditional Chinese medicine systems pharmacology database and analysis platform (TCMSP) and encyclopedia of traditional Chinese medicine (ETCM). The common targets shared by Tusizi prescription and diminished ovarian reserve (DOR) were selected and imported into search tool for the retrieval of interacting genes/proteins (STRING) to construct a protein-protein interaction (PPI) network and into gene function annotation database (DAVID) for gene ontology (GO) analysis. The CCK-8 assay was used to measure the viability of ovarian germline stem cells treated with hyperoside. The CCK-8 assay, 5-ethynyl-2'-deoxyuridine (EdU) staining, terminal-deoxynucleoitidyl transferase mediated nick-end labeling (TUNEL), and enzyme-linked immunosorbent assay (ELISA) were employed to examine the proliferation, apoptosis, and estradiol (E2) secretion of KGN cells treated with the water extract 11 of Tusizi prescription (Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 4∶4∶2∶1∶1) and the optimal prescription screened by uniform design. On this basis, the optimal prescription composition for maximizing the effect on ovarian reserve function was determined and preliminary insights into the underlying mechanisms of this prescription were gained. ResultsA total of 147 common targets were obtained from 278 targets of Tusizi prescription and 1 721 targets of DOR. GO analysis revealed 194 biological processes, primarily involving cellular responses to exogenous compound stimuli, negative regulation of apoptotic process, and positive regulation of cell proliferation. It identified 84 cellular components, including cell membrane, mitochondria, and neuronal cell body, as well as 144 molecular functions such as enzyme binding, estrogen response element binding, and nuclear estrogen receptor binding. The multivariate regression analysis revealed that when Tusizi prescription was composed of Cuscutae Semen, Lycii Fructus, Dioscoreae Rhizoma, Poria, and Nelumbinis Semen in a ratio of 27∶30∶17∶12∶14, the water extract of Tusizi prescription had the best effect of enhancing the viability of KGN cells. CCK-8 results showed that compared with the normal group, the hyperoside group demonstrated increased viability of ovarian germline stem cells (P<0.01). The CCK-8, EdU, and ELISA results showed that compared with the normal group, the optimal prescription screened by uniform design and the water extract 11 of Tusizi prescription increased the proliferation and reduced the apoptosis of KGN cells (P<0.05, P<0.01). ELISA results showed that compared with the normal group, the water extract 11 of Tusizi prescription promoted the E2 secretion of KGN cells (P<0.05), while the optimal prescription screened by uniform design had no significant effect on the E2 secretion. ConclusionBoth the water extract 11 of Tusizi prescription (Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 4∶4∶2∶1∶1) and the optimal prescription screened by uniform design (Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 27∶30∶17∶12∶14) can improve the ovarian reserve function, and the former has better effect. Tusizi prescription can modulate biological processes (such as cell proliferation and apoptosis) and molecular functions (such as enzyme binding and estrogen response element binding) through active components like hyperoside to promote the proliferation and E2 secretion and inhibit the apoptosis of KGN cells, thereby protecting the ovarian reserve function.
10.Optimization and Mechanism Exploration of Tusizi Prescription for Ovarian Reserve Function Based on Uniform Design Method
Yuan LI ; Hanqian DU ; Jiashan LI ; Li GUO ; Zehui LI ; Na LIN ; Ying XU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(7):53-62
ObjectiveTo optimize Tusizi prescription for ovarian reserve function based on the uniform design method combined with in vitro experiments and explore the underlying mechanisms of this prescription. MethodsThe uniform design method was adopted to design a 5-factor 11-level experiment on the water extract of Tusizi prescription. The cell-counting kit-8 (CCK-8) assay was employed to measure the viability of human ovarian granulosa cells (KGN cells) treated with Tusizi prescription extracts 1-11, and multivariate regression analysis was performed to determine the optimal herb ratio in this prescription. The potential targets of active ingredients in the prescription were retrieved from traditional Chinese medicine systems pharmacology database and analysis platform (TCMSP) and encyclopedia of traditional Chinese medicine (ETCM). The common targets shared by Tusizi prescription and diminished ovarian reserve (DOR) were selected and imported into search tool for the retrieval of interacting genes/proteins (STRING) to construct a protein-protein interaction (PPI) network and into gene function annotation database (DAVID) for gene ontology (GO) analysis. The CCK-8 assay was used to measure the viability of ovarian germline stem cells treated with hyperoside. The CCK-8 assay, 5-ethynyl-2'-deoxyuridine (EdU) staining, terminal-deoxynucleoitidyl transferase mediated nick-end labeling (TUNEL), and enzyme-linked immunosorbent assay (ELISA) were employed to examine the proliferation, apoptosis, and estradiol (E2) secretion of KGN cells treated with the water extract 11 of Tusizi prescription (Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 4∶4∶2∶1∶1) and the optimal prescription screened by uniform design. On this basis, the optimal prescription composition for maximizing the effect on ovarian reserve function was determined and preliminary insights into the underlying mechanisms of this prescription were gained. ResultsA total of 147 common targets were obtained from 278 targets of Tusizi prescription and 1 721 targets of DOR. GO analysis revealed 194 biological processes, primarily involving cellular responses to exogenous compound stimuli, negative regulation of apoptotic process, and positive regulation of cell proliferation. It identified 84 cellular components, including cell membrane, mitochondria, and neuronal cell body, as well as 144 molecular functions such as enzyme binding, estrogen response element binding, and nuclear estrogen receptor binding. The multivariate regression analysis revealed that when Tusizi prescription was composed of Cuscutae Semen, Lycii Fructus, Dioscoreae Rhizoma, Poria, and Nelumbinis Semen in a ratio of 27∶30∶17∶12∶14, the water extract of Tusizi prescription had the best effect of enhancing the viability of KGN cells. CCK-8 results showed that compared with the normal group, the hyperoside group demonstrated increased viability of ovarian germline stem cells (P<0.01). The CCK-8, EdU, and ELISA results showed that compared with the normal group, the optimal prescription screened by uniform design and the water extract 11 of Tusizi prescription increased the proliferation and reduced the apoptosis of KGN cells (P<0.05, P<0.01). ELISA results showed that compared with the normal group, the water extract 11 of Tusizi prescription promoted the E2 secretion of KGN cells (P<0.05), while the optimal prescription screened by uniform design had no significant effect on the E2 secretion. ConclusionBoth the water extract 11 of Tusizi prescription (Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 4∶4∶2∶1∶1) and the optimal prescription screened by uniform design (Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 27∶30∶17∶12∶14) can improve the ovarian reserve function, and the former has better effect. Tusizi prescription can modulate biological processes (such as cell proliferation and apoptosis) and molecular functions (such as enzyme binding and estrogen response element binding) through active components like hyperoside to promote the proliferation and E2 secretion and inhibit the apoptosis of KGN cells, thereby protecting the ovarian reserve function.

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