1.Effects of different electromagnetic stimulation protocols on hand function and activities of daily living for stroke patients: a network meta-analysis
Shan LIU ; Kunyao XU ; Zhimeng ZHOU ; Lingyun ZHOU ; Mao ZENG ; Sha HE ; Luoyang DAI ; Ting QIN ; Qinglin HU ; Jiahai YANG ; Rui LI
Chinese Journal of Rehabilitation Theory and Practice 2026;32(8):916-926
ObjectiveTo systematically compare the effects of diverse electromagnetic stimulation protocols on hand function and activities of daily living (ADL) in stroke patients using network meta-analysis (NMA), and screen the optimal stimulation strategy for post-stroke hand dysfunction rehabilitation. MethodsNMA was performed in accordance with the PRISMA-NMA statement. Databases of CNKI, Wanfang data, VIP, CBM, PubMed, Web of Science and Embase were searched from database inception to April, 2026 for eligible randomized controlled trials (RCTs) comparing different electromagnetic stimulation protocols for post-stroke hand impairment. The Cochrane Risk of Bias tool and PEDro scale were used to assess methodological quality of included studies, while the GRADE system was adopted to grade evidence quality of primary outcomes of Fugl-Meyer Assessment-Upper Extremities (FMA-UE) and modified Barthel Index (MBI). After literature screening, data extraction and bias evaluation, RevMan 5.4 and Stata 17.0 were used for meta-analysis and NMA. ResultsA total of 22 RCT involving 17 electromagnetic stimulation protocols were finally included. The PEDro scores ranged from six to ten. The top five protocols for improving FMA-UE score were repetitive peripheral magnetic stimulation (rPMS) + repetitive transcranial magnetic stimulation (rTMS), intermittent theta-burst stimulation (iTBS) + rPMS, iTBS + transcranial direct current stimulation (tDCS), transcutaneous electrical acupoint stimulation (TEAS) and contralaterally controlled functional electrical stimulation (CCFES). tDCS was the most effective on MBI, followed by TEAS and iTBS+rPMS. Evidence quality for FMA-UE and MBI was moderate. Funnel plots suggested low risk of publication bias, and consistency tests verified good coherence between direct and indirect comparisons in network models. ConclusionElectromagnetic stimulation significantly facilitates upper limb and hand functional recovery, and improves ADL in stroke patients. The combined central-peripheral magnetic stimulation protocol may be the most effective on motor rehabilitation, whereas tDCS on ADL.
2.Exploring Mechanism of Luoshi Neiyi Prescription in Treating Endometriosis Based on Ferroptosis and Serum Metabolomics
Haixia PAN ; Yingqiao ZHONG ; Ting MAO ; Ziyi DENG ; Meilin WU ; Lei HUANG ; Siyang CHEN ; Yong GUO ; Ying ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(20):201-212
ObjectiveThis study aimed to investigate the mechanism by which Luoshi Neiyi prescription treats endometriosis (EMs) through regulating ferroptosis, and to screen key metabolites and analyze their association with ferroptosis. MethodsClinical samples of normal endometrium from patients without EMs and eutopic and ectopic endometrium from EMs patients (10 cases each) were collected and divided into control group, eutopic group, and EMs group. Hematoxylin-eosin (HE) staining was performed to observe ectopic lesions of EMs. Immunohistochemistry was used to detect the expression of solute carrier family 7 member 11 (SLC7A11) and glutathione peroxidase 4 (GPX4). Enzyme-linked immunosorbent assay (ELISA) was adopted to determine the levels of malondialdehyde (MDA), ferrous ion (Fe2+), GPX4 and glutathione (GSH) in endometrial tissues, as well as serum levels of Fe2+, GPX4 and GSH. Real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of SLC7A11, GPX4, transferrin receptor (TFR) and ferritin heavy chain 1 (FTH1). In the in vitro experiment, primary stromal cells were isolated from ectopic lesions of EMs patients. Cell counting kit-8 (CCK-8) was used to determine the optimal concentration of drug-containing serum for intervention. The level of reactive oxygen species (ROS) was measured, and Real-time PCR was applied to detect ferroptosis-related indicators. In the animal experiments, an EM rat model was established, and the rats were randomly assigned to the sham operation group, EMs group, low-dose Luoshi Neiyi Formula group (7.87 g·kg-1), high-dose Luoshi Neiyi prescription group (15.74 g·kg-1), and danazol group (42 mg·kg-1). Untargeted metabolomics detection and pathway enrichment analysis were conducted on serum samples from patients and rats. Spearman correlation analysis was performed to assess the relationship between differential metabolites and ferroptosis indicators. The correlations between differential metabolites in patient endometrium and serum and key ferroptosis indicators (GPX4, Fe2+, MDA, GSH) as well as ferroptosis-related mRNAs (GPX4, SLC7A11, FTH1) were analyzed, and correlation heatmaps were generated accordingly. ResultsCompared with normal eutopic endometrium, ectopic lesions in EMs patients showed glandular disorganization and stromal fibrosis. In ectopic endometrium, the contents of MDA, ROS, and Fe2+ decreased, while GPX4 level increased, and the mRNA expression of SLC7A11 and GPX4 was upregulated (P<0.05, P<0.01). In serum, the levels of GPX4 and Fe2+ were elevated, whereas the GSH level declined, suggesting abnormalities in ferroptosis-related pathways in ectopic lesions (P<0.05, P<0.01). After intervention with Luoshi Neiyi prescription-containing serum, the intracellular ROS level in ectopic endometrial stromal cells was elevated, the mRNA expression of SLC7A11 and GPX4 was downregulated, and TFR mRNA expression was upregulated (P<0.05, P<0.01). Metabolomics analysis revealed 1104 and 198 differential metabolites in EMs patients and EMs rats, respectively, compared with their corresponding control groups, and both low-dose and high-dose Luoshi Neiyi prescription were found to regulate this metabolic disturbance, with the core regulatory pathways mainly involving arginine and proline metabolism. Correlation analysis showed that the glycerophospholipids including PI(16∶0/17∶0) and PI[18∶2(9Z,12Z)] were negatively correlated with GPX4 and positively correlated with MDA, while 17α-hydroxyprogesterone was positively correlated with GPX4, SLC7A11, and FTH1 q<0.05). ConclusionLuoshi Neiyi prescription may systematically ameliorate disease-associated metabolic dysregulation via modulation of the serum arginine and proline metabolism pathway, and may regulate ferroptosis in ectopic lesions through a mechanism potentially linked to the serum glycerophospholipid and steroid metabolism pathways. Collectively, these findings provide experimental evidence for the clinical application of Luoshi Neiyi prescription.
3.Exploring Mechanism of Luoshi Neiyi Prescription in Treating Endometriosis Based on Ferroptosis and Serum Metabolomics
Haixia PAN ; Yingqiao ZHONG ; Ting MAO ; Ziyi DENG ; Meilin WU ; Lei HUANG ; Siyang CHEN ; Yong GUO ; Ying ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(20):201-212
ObjectiveThis study aimed to investigate the mechanism by which Luoshi Neiyi prescription treats endometriosis (EMs) through regulating ferroptosis, and to screen key metabolites and analyze their association with ferroptosis. MethodsClinical samples of normal endometrium from patients without EMs and eutopic and ectopic endometrium from EMs patients (10 cases each) were collected and divided into control group, eutopic group, and EMs group. Hematoxylin-eosin (HE) staining was performed to observe ectopic lesions of EMs. Immunohistochemistry was used to detect the expression of solute carrier family 7 member 11 (SLC7A11) and glutathione peroxidase 4 (GPX4). Enzyme-linked immunosorbent assay (ELISA) was adopted to determine the levels of malondialdehyde (MDA), ferrous ion (Fe2+), GPX4 and glutathione (GSH) in endometrial tissues, as well as serum levels of Fe2+, GPX4 and GSH. Real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of SLC7A11, GPX4, transferrin receptor (TFR) and ferritin heavy chain 1 (FTH1). In the in vitro experiment, primary stromal cells were isolated from ectopic lesions of EMs patients. Cell counting kit-8 (CCK-8) was used to determine the optimal concentration of drug-containing serum for intervention. The level of reactive oxygen species (ROS) was measured, and Real-time PCR was applied to detect ferroptosis-related indicators. In the animal experiments, an EM rat model was established, and the rats were randomly assigned to the sham operation group, EMs group, low-dose Luoshi Neiyi Formula group (7.87 g·kg-1), high-dose Luoshi Neiyi prescription group (15.74 g·kg-1), and danazol group (42 mg·kg-1). Untargeted metabolomics detection and pathway enrichment analysis were conducted on serum samples from patients and rats. Spearman correlation analysis was performed to assess the relationship between differential metabolites and ferroptosis indicators. The correlations between differential metabolites in patient endometrium and serum and key ferroptosis indicators (GPX4, Fe2+, MDA, GSH) as well as ferroptosis-related mRNAs (GPX4, SLC7A11, FTH1) were analyzed, and correlation heatmaps were generated accordingly. ResultsCompared with normal eutopic endometrium, ectopic lesions in EMs patients showed glandular disorganization and stromal fibrosis. In ectopic endometrium, the contents of MDA, ROS, and Fe2+ decreased, while GPX4 level increased, and the mRNA expression of SLC7A11 and GPX4 was upregulated (P<0.05, P<0.01). In serum, the levels of GPX4 and Fe2+ were elevated, whereas the GSH level declined, suggesting abnormalities in ferroptosis-related pathways in ectopic lesions (P<0.05, P<0.01). After intervention with Luoshi Neiyi prescription-containing serum, the intracellular ROS level in ectopic endometrial stromal cells was elevated, the mRNA expression of SLC7A11 and GPX4 was downregulated, and TFR mRNA expression was upregulated (P<0.05, P<0.01). Metabolomics analysis revealed 1104 and 198 differential metabolites in EMs patients and EMs rats, respectively, compared with their corresponding control groups, and both low-dose and high-dose Luoshi Neiyi prescription were found to regulate this metabolic disturbance, with the core regulatory pathways mainly involving arginine and proline metabolism. Correlation analysis showed that the glycerophospholipids including PI(16∶0/17∶0) and PI[18∶2(9Z,12Z)] were negatively correlated with GPX4 and positively correlated with MDA, while 17α-hydroxyprogesterone was positively correlated with GPX4, SLC7A11, and FTH1 q<0.05). ConclusionLuoshi Neiyi prescription may systematically ameliorate disease-associated metabolic dysregulation via modulation of the serum arginine and proline metabolism pathway, and may regulate ferroptosis in ectopic lesions through a mechanism potentially linked to the serum glycerophospholipid and steroid metabolism pathways. Collectively, these findings provide experimental evidence for the clinical application of Luoshi Neiyi prescription.
4.Rare relapsed TTMV::RARA fusion gene-positive pediatric acute promyelocytic leukemia: report of 1 case and review of literature
Ting ZHAO ; Luyue DING ; Jianwen ZHOU ; Pengkai FAN ; Mingfa GUO ; Chunxiang LI ; Yutai SU ; Yanna MAO ; Ping MA
Journal of Leukemia & Lymphoma 2025;34(8):494-497
Objective:To improve the understanding of pediatric acute promyelocytic leukemia with TTMV::RARA fusion gene positive caused by torque teno mini virus (TTMV).Methods:A retrospective analysis was conducted on the clinical data of a patient with relapsed TTMV::RARA fusion gene-positive acute promyelocytic leukemia who was admitted to Children's Hospital Affiliated to Zhengzhou University in July 2024, and literature review was conducted.Results:The patient was a girl with the age of 5 years and 7 months. She presented with joint pain and fever. Combined with bone marrow cell morphology and whole transcriptome sequencing, she was diagnosed with TTMV::RARA fusion gene-positive acute promyelocytic leukemia. After induction therapy with regimens such as retinoic acid +daunorubicin+cytarabine and retinoic acid+venetoclax+homoharringtonine, the joint pain was relieved, but the primary disease did not improve. Subsequently, there was no regular treatment. One year later, the disease recurred and was complicated with severe infection. Her condition improved following anti-infection and induction therapy.Conclusions:TTMV::RARA fusion gene-positive pediatric acute promyelocytic leukemia is a special type of acute promyelocytic leukemia caused by the insertion of viral sequences from TTMV infection. It is rare in clinical practice and difficult to treat, and the overall prognosis may be poor.
5.Pathogenicity and multiple detection methods for infectious diarrhea in children under 5 years old in Zhongshan City from 2023 to 2024
Wuyang SHI ; Ting OUYANG ; Shuhuan YANG ; Yunxia MAO ; Yanheng WU ; Bo HE
Chinese Journal of Infection Control 2025;24(10):1402-1408
Objective To analyze the pathogen spectrum characteristics of infectious diarrhea in children under 5 years old in Zhongshan City from 2023 to 2024,and evaluate the application value of multiple detection technique in monitoring diarrhea syndrome.Methods Diarrhea specimens from hospitalized children under 5 years old in 4 senti-nel hospitals in Zhongshan City from 2023 to 2024 were collected,Luminex? multi-pathogen detection kit(GPP)was used for screening 16 types of pathogens,and fluorescence quantitative polymerase chain reaction(qPCR)was simultaneously used to verify the consistency of detection results of 5 diarrhea virus.Results A total of 578 dia-rrhea specimens were collected,and the positive detection rate of pathogens was 67.13%(n=388).The positive detection rate of viral pathogens was 38.24%,mainly norovirus(21.63%),rotavirus A(10.90%),and sapovirus(4.67%).The positive detection rate of bacterial pathogens was 41.00%,mainly Salmonella spp.(19.55%),Clostridioides di f ficile toxin A/B(14.71%),and Campylobacter sp p.(8.82%).Cryptosporidium,Entamoeba histolytica,and Giardia were not detected.The consistency between GPP and qPCR in detecting viral pathogens reached 95.16%,with a Kappa value of 0.897(x2=465.36,P<0.001).Conclusion The main pathogens causing diarrhea in children in Zhongshan City are norovirus,Salmonella,and Clostridioides toxin A/B.GPP technique can efficiently construct a multi-pathogen spectrum,and provide reliable technical support for optimizing the monitoring system of diarrhea syndrome.
6.Expression levels of miR-127-5p,miR-127 and miR-155 in elderly patients with pneumonia infection by multidrug-resistant bacteria and their correlation with immune-related cytokines
Yanqing MAO ; Ting FU ; Juan WANG ; Ya'nan WANG ; Jie LI
The Journal of Practical Medicine 2025;41(3):385-390
Objective To explore the expression levels of ribonucleic acid(miR)-127-5p,miR-127 and miR-155 in elderly patients with pneumonia infection by multidrug-resistant bacteria and their correlation with immune-related cytokines.Methods 65 elderly patients with pneumonia infection by multidrug-resistant bacteria,80 healthy physical examination persons and 80 elderly patients with common pneumonia in our hospital from June 2020 to June 2022 were retrospectively selected as the observation group,the control group A and the control group B,respectively.The expression levels of miR-127-5p,miR-127,miR-155,interferon-γ(IFN-γ),interleu-kin(IL-4),IL-17,and transforming growth factor-β(TGF-β)in serum samples were detected and compared among the three groups.Pearson correlation analysis analyzed the correlation between miR-127-5p,miR-127,miR-155 and IFN-γ,IL-4,IL-17,TGF-β,and the Receiver operating characteristic curve(ROC)analyzed the diagnostic value of miR-127-5p,miR-127,miR-155 alone or in combination.Results The serum levels of miR-127-5p and IFN-γ in the control group B and the observation group were lower than those in the control group A,and the both in the observation group were lower than the both in the control group B(P<0.05).The serum levels of miR-127,miR-155,IL-4,IL-17,and TGF-β in the control group B and observation group were higher than those in the control group A,and those in the observation group were higher than those in the control group B(P<0.05).There was a positive correlation between serum miR-127-5p and serum IFN-γ levels(r=0.463,P<0.05),serum miR-127 and IL-4,IL-17 or TGF-β levels(r=0.306,0.273 or 0.330,P<0.05),and serum miR-155 and IL-4,IL-17 or TGF-β levels(r=0.410,0.453 or 0.385,P<0.05),and as well as a negative correlation between serum miR-127-5p and IL-4,IL-17 or TGF-β(r=-0.378,-0.396 or-0.408,P<0.05),serum miR-127 or miR-155 and IFN-γ levels(r=-0.267 or-0.513,P<0.05).Serum miR-127-5p,miR-127 and miR-155 expression levels alone and in combination have a certain diagnostic value in elderly patients with pneumonia infec-tion by multi-drug resistant bacteria,and the area under ROC curve(AUC)values were 0.811,0.733,0.817 and 0.906,and the sensitivities were 90.77%,60.00%,67.69%,84.62%,and the specificities were 66.87%,80.62%,83.75%,83.18%.Conclusion MiR-127-5p,miR-127 and miR-155 were differentially expressed in the elderly patients with pneumonia infection by multi-drug resistant bacteria,and were correlated with immune-related cytokines to a certain extent,and the combination of the three has a good diagnostic value,thus may providing a new target for clinical diagnosis of this disease.
7.Effects of methyl oxofulnonone A ameliorates hyperglucose-induced H9c2 cardiomyocyte injury
Ming-hui OUYANG ; Xiao-tong GUO ; Ting LIU ; Li-ying CHENG ; Zi-chang NIU ; Hao-ping MAO
The Chinese Journal of Clinical Pharmacology 2025;41(1):45-49
Objective To investigate the effect and mechanism of methyl oxofulnonone A(META)on high glucose(HG)-induced H9c2 cell injury.Methods H9c2 cells were divided into control group(normal culture),model group(55 mmol·L-1 glucose)and experimental-L,-M,-H groups(55 mmol·L-1 glucose+12.5,25.0,50.0 μmol·L-1 META).Cell viability was detected by cell counting kit-8;intracellular reactive oxygen species(ROS)level was detected by DCFH-DA fluorescent probe;intracellular adenosine triphosphate(ATP)content was detected by luciferase;and autophagy-related protein expression was detected by Western blotting.Results The optical density values of 72-hour cells in the control group,model group and experimental-M,-H groups were 0.91±0.03,0.61±0.01,0.69±0.02 and 0.72±0.03;the ROS levels were(40.75±1.53)%,(43.73±1.30)%,(30.87±1.27)%and(28.28±1.43)%;the ATP contents were(8.16±0.71),(4.03±0.29),(5.29±0.31)and(5.83±0.31)nmol·mg-1;the relative expression levels of autophagy-related gene 5 protein were 1.05±0.06,1.46±0.09,0.98±0.11 and 0.89±0.09;the relative expression levels of ubiquitin-binding protein were 1.05±0.10,0.55±0.13,0.89±0.04 and 0.98±0.04;the ratios of microtubule-associated protein 1 light chain 3 11/Ⅰ protein were 1.09±0.09,1.82±0.05,1.67±0.29 and 1.09±0.15,respectively.Among the above indicators,there were statistically significant differences between the model group and the control and experimental-M,-H groups(P<0.05,P<0.01).Conclusion META significantly ameliorates H9c2 cardiomyocyte damage caused by high glucose,ameliorates oxidative stress,protects mitochondrial respiration and inhibits autophagy.
8.Application of wide awake local anesthesia no tourniquet technique in locking of metacarpophalangeal joint with extension lag surgery
Guo-tao YANG ; Hua-jie MAO ; Zhi-jun CHEN ; Xu ZHANG ; Huan HU ; Qi-ting JIANG
Journal of Regional Anatomy and Operative Surgery 2025;34(9):836-839
Objective To explore the application value of wide awake local anesthesia no tourniquet(WALANT)technique in outpatient surgery for locking of metacarpophalangeal joint with extension lag.Methods The clinical data of 6 patients with locking of meta-carpophalangeal joint with extension lag in Danyang People's Hospital from January 2019 to October 2023 were retrospectively analyzed.The patients were received outpatient surgery under the WALLANT technique for release,and lidocaine mixed solution containing 1∶100 000 epinephrine was injected into the proximal midpoint of the volar projection of the metacarpophalangeal joint.The joint was exposed with a volar or dorsal finger web incision to determine the unrestricted structure,and the collateral ligament and paralateral collateral ligament with high tension were cut off.The intraoperative blood loss,postoperative incision healing and complications were recorded.Visual analogue scale(VAS)was used to evaluate the intraoperative pain,and the range of motion of metacarpophalangeal joint and total active movement(TAM)of finger joint were observed during postoperative follow-up.Results The incision of patients were healed successfully in the first phase after surgery,without wound necrosis.The anesthesia effects of patients were all satisfied and the operation was successfully completed.The VAS score was less than 3 points and there was only a small amount of bleeding during the operation.The recovery of joint flexion and extension movements could be observed during the operation,and the TAM score after the operation was 20 points.No significant change was found on the range of motion of metacarpophalangeal joint or TAM of finger joint between the injured finger and the corresponding healthy finger(P>0.05).Conclusion WALANT technique for locking of metacarpophalangeal joint with extension lag surgery has good anesthesia effect,less bleeding in the incision,and clear vision of the surgery.It can avoid vascular and nerve injuries,observe the recovery of joint activities during the operation and relieve pain of patients,which is conducive to outpatient surgery and saving medical and social resources at the same time.
9.Underlying target of bullatine A in treating rheumatoid arthritis based on LiP-SMap drug target proteomics
Hao-hong ZHANG ; Nan-ting ZOU ; Chun-fei ZHANG ; Qing-yan MO ; Ming-qian JU ; Xiao-hong LI ; Shuai LIU ; Mao-kui HUANG ; Hong-yun WANG ; Chun-ping WAN
Chinese Pharmacological Bulletin 2025;41(6):1072-1078
Aim To identify the underlying target of bullatine A(BA)against rheumatoid arthritis(RA)u-sing limited proteolysis-small molecule mapping(LiP-SMap)drug target proteomics and to provide a scientif-ic basis for clinical application of Aconiti brachypodi Radix in the treatment of RA.Methods LiP-SMap drug target proteomics was employed to perform bioin-formatics analysis for comparing and validating the dif-ferential protein expression after BA intervention.A collagen-induced arthritis(CIA)model was estab-lished in DBA/1 mice using bovine type Ⅱ collagen.The mice were then divided into the CIA model group,methotrexate-positive control group(MTX group),and BA groups(10 mg·kg-1 and 20 mg·kg-1)based on their clinical scores.After drug intervention,the thera-peutic efficacy against RA was assessed by joint index scores and foot thickness measurements.Histopatholog-ical changes in the arthritic joints of CIA mice were e-valuated using hematoxylin and eosin(HE)staining.Enzyme-linked immunosorbent assay(ELISA)was employed to detect inflammatory cytokines interleukin-17(IL-17)and total IgG and IgG3 anti-collagen-spe-cific antibodies levels from the serum of CIA mice.Flow cytometry was used to detect the expression levels of intracellular Th17 cells(IL-17+CD4+T cells)and Th1 cells(IFN-γ+CD4+T cells).Fluorescent quanti-tative PCR was performed to detect the expression of genes related to differential proteins.Results The proteomic analysis identified Serpinb1a as a protein with strong binding affinity to BA,and KEGG enrich-ment analysis indicated IL-17 signaling pathway was a crucial pathway of BA in against RA.BA treatment significantly reduced clinical scores and foot thickness,improved local arthritis symptoms in CIA mice,and al-leviated inflammatory cell infiltration into arthritic joints(P<0.05).Differential protein validation re-sults showed that BA had strong affinity with Serpinb1a(-5.92 kJ·mol-1)and downregulated the expres-sion of Serpinb1a mRNA.Furthermore,the administra-tion of BA markedly reduced serum IL-17 A levels from CIA mice,inhibited the expression of intracellular IL-17 A and IFN-γ cytokines in splenic CD4+T cells(P<0.05),and significantly downregulated the transcrip-tional expression of IL-17F(P<0.05).Conclusion BA exhibits therapeutic effects on collagen-induced arthritis,and its mechanism of action may involve the regulation of Serpinb1a and the IL-17 signaling path-way.
10.Icariin improves injury of tight junctional function by regulating balance of mTORC1 and mTORC2 in testicular Sertoli cells in naturally aging mice
Yao-ting CHENG ; Chang-cheng ZHANG ; Guo-qing FU ; Tan WANG ; Jian-min MAO ; Jian-ming SUN ; Hai-xia ZHAO
Chinese Pharmacological Bulletin 2025;41(6):1091-1098
Aim To explore the protective effect of icariin on the damage of tight junctional function of Sertoli cells in naturally aging mice and the related mechanism.Methods 15-month-old C57BL/6J male mice were randomly divided into three groups:aging model group,low-dose and high-dose icariin treatment group(5 and 20 mg·kg-1).Another 1-month-old C57BL/6J male mice were considered as adult control group(n=10).The mice in adult control group and aging model group were given the vehicle(0.5%sodi-um carboxymethyl cellulose solution)by intragastric administration,while the mice in icariin-treated groups were given different concentrations of icariin,respec-tively.After continuous administration of icariin for three months,the testes and epididymis were immedi-ately removed,weighed,and the organ index was calcu-lated.Sperm viability and sperm concentration in epi-didymis were measured.The morphological changes of testes were observed by HE staining.The ultrastructur-al changes of tight junctions of Sertoli cells were ob-served by transmission electron microscopy.The ex-pression levels of tight junction-related proteins ZO-1,Occludin,and Claudin11 of testicular Sertoli cells were detected by Western blot.The expression and localiza-tion of ZO-1,Occludin,Raptor,Rictor,p-70S6K,and p-rps6 were detected by immunofluorescence.Results Compared with the aging model group,icariin signifi-cantly increased testicular weight and its index,and ep-ididymal index,improved sperm viability and increased sperm concentration in naturally aging mice.In addi-tion,icariin improved the degeneration of testicular morphology and the damage of ultrastructure of Sertoli cell tight junction with aging.Furthermore,Western blot results showed that icariin up-regulated the expres-sion of ZO-1 and Occludin,but had no significant effect on the expression of Claudin 11.Immunofluorescence assay showed that icariin up-regulated the expression of Rictor,and down-regulated the expression of p-70S6K,p-rps6 and Raptor.Conclusions Icariin improves the tight junction damage of Sertoli cells in naturally aging mice,and its mechanism may be related to restoring the balance between mTORC1 and mTORC2.

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