1.Molecular mechanisms and diagnostic value of seminal plasma exosomal miR-26a-5p targeting PTEN in idiopathic teratozoospermia
Chunhui LIU ; Ruipeng WU ; Zhiqiang WANG ; Wensheng SHAN ; Shaojun LI
The Journal of Practical Medicine 2025;41(20):3256-3266
Objective This study aims to elucidate the functional mechanism through which seminal plasma exosomal miR-26a-5p and its target genes contribute to idiopathic teratozoospermia,with emphasis on their regulatory pathways in sperm morphogenesis defects,thereby establishing a theoretical foundation for novel diagnostic markers and targeted therapies.Methods A case-control study design was adopted,enrolling 154 male subjects(84 in the teratozoospermia group[TZS]and 70 in the normal control group[NC]).Seminal plasma exosomes were isolated to screen differentially expressed miRNAs,followed by prediction and analysis of target genes and signaling pathways.Dual-luciferase reporter gene assays were employed to validate the direct binding of miR-26a-5p to PTEN.Quantitative real-time PCR and Western blot were used to measure miRNA and protein expression levels.Spearman correlation analysis evaluated relationships between miR-26a-5p,PTEN,and sperm morphological parameters,while ROC curve analysis assessed diagnostic efficacy.Results Seminal plasma exosomal miRNA sequencing identified 11 differentially expressed miRNAs,with miR-26a-5p significantly upregulated in the TZS group(P<0.05)and negatively correlated with the percentage of normal sperm morphology in TZS(r=-0.762,P<0.05).PTEN was confirmed as a direct target of miR-26a-5p,with its expression significantly reduced in the TZS group(P<0.05)and positively correlated with normal sperm morphology(r=0.821,P<0.05).A negative correlation was observed between miR-26a-5p and PTEN(r=-0.878,P<0.05).Dual-luciferase assays demonstrated that miR-26a-5p specifically inhibited PTEN 3'UTR activity(45%reduction in luciferase activity,P<0.05).Functional enrichment analysis revealed that the miR-26a-5p-PTEN axis exacerbates oxidative stress,DNA damage,and abnormal spermatocyte division by regulating the PI3K/AKT/mTOR,p53,Wnt/β-catenin,and NF-κB pathways.ROC analysis showed diagnostic efficacy for teratozoospermia with AUCROC values of 0.785(sensitivity 78.9%,specificity 71.3%)for miR-26a-5p and 0.754(sensitivity 73.3%,specificity 75.1%)for PTEN.Conclusions miR-26a-5p suppresses PTEN through targeted inhibition,activating PI3K/AKT/mTOR signaling pathways while impairing DNA repair functions.This cascade leads to accumulated oxidative stress and sperm morphological abnormalities.The combined pattern of elevated seminal plasma exosomal miR-26a-5p expression and reduced PTEN levels demonstrates diagnostic potential for idiopathic teratozoospermia,with its molecular mechanism providing a theoretical foundation for biomarker development.Targeted silencing of miR-26a-5p,either alone or in combination with AKT/mTOR inhibitors and antioxidant therapy,may represent a novel strategy for improving sperm morphology.
2.Molecular mechanisms and diagnostic value of seminal plasma exosomal miR-26a-5p targeting PTEN in idiopathic teratozoospermia
Chunhui LIU ; Ruipeng WU ; Zhiqiang WANG ; Wensheng SHAN ; Shaojun LI
The Journal of Practical Medicine 2025;41(20):3256-3266
Objective This study aims to elucidate the functional mechanism through which seminal plasma exosomal miR-26a-5p and its target genes contribute to idiopathic teratozoospermia,with emphasis on their regulatory pathways in sperm morphogenesis defects,thereby establishing a theoretical foundation for novel diagnostic markers and targeted therapies.Methods A case-control study design was adopted,enrolling 154 male subjects(84 in the teratozoospermia group[TZS]and 70 in the normal control group[NC]).Seminal plasma exosomes were isolated to screen differentially expressed miRNAs,followed by prediction and analysis of target genes and signaling pathways.Dual-luciferase reporter gene assays were employed to validate the direct binding of miR-26a-5p to PTEN.Quantitative real-time PCR and Western blot were used to measure miRNA and protein expression levels.Spearman correlation analysis evaluated relationships between miR-26a-5p,PTEN,and sperm morphological parameters,while ROC curve analysis assessed diagnostic efficacy.Results Seminal plasma exosomal miRNA sequencing identified 11 differentially expressed miRNAs,with miR-26a-5p significantly upregulated in the TZS group(P<0.05)and negatively correlated with the percentage of normal sperm morphology in TZS(r=-0.762,P<0.05).PTEN was confirmed as a direct target of miR-26a-5p,with its expression significantly reduced in the TZS group(P<0.05)and positively correlated with normal sperm morphology(r=0.821,P<0.05).A negative correlation was observed between miR-26a-5p and PTEN(r=-0.878,P<0.05).Dual-luciferase assays demonstrated that miR-26a-5p specifically inhibited PTEN 3'UTR activity(45%reduction in luciferase activity,P<0.05).Functional enrichment analysis revealed that the miR-26a-5p-PTEN axis exacerbates oxidative stress,DNA damage,and abnormal spermatocyte division by regulating the PI3K/AKT/mTOR,p53,Wnt/β-catenin,and NF-κB pathways.ROC analysis showed diagnostic efficacy for teratozoospermia with AUCROC values of 0.785(sensitivity 78.9%,specificity 71.3%)for miR-26a-5p and 0.754(sensitivity 73.3%,specificity 75.1%)for PTEN.Conclusions miR-26a-5p suppresses PTEN through targeted inhibition,activating PI3K/AKT/mTOR signaling pathways while impairing DNA repair functions.This cascade leads to accumulated oxidative stress and sperm morphological abnormalities.The combined pattern of elevated seminal plasma exosomal miR-26a-5p expression and reduced PTEN levels demonstrates diagnostic potential for idiopathic teratozoospermia,with its molecular mechanism providing a theoretical foundation for biomarker development.Targeted silencing of miR-26a-5p,either alone or in combination with AKT/mTOR inhibitors and antioxidant therapy,may represent a novel strategy for improving sperm morphology.
3.Research progress on the intervention of traditional Chinese medicine monomer and formula in the mTOR signaling pathway for the treatment of osteoporosis
Yan CHEN ; Liufeng SHAN ; Wensheng CHE ; Yujun LIU ; Zhiqi LIU ; Xiaoxue SHI ; Zhihai FENG
China Pharmacy 2023;34(23):2934-2938
Osteoporosis is an important cause of bone weakness and susceptibility to fractures. Anti-osteoporosis drugs of Western medicine cannot reverse its progression, and can only reduce the loss of bone density; long-term use of them is accompanied by certain adverse reactions. Traditional Chinese medicine focuses on syndrome differentiation and holistic approach, which can make up for the shortcomings of Western medicine’s treatment. The mammalian target of rapamycin (mTOR) signaling pathway is involved in the growth, proliferation, and differentiation of bone cells, and is closely related to the occurrence and development of osteoporosis. In recent years, various traditional Chinese medicine monomers (such as flavonoids, polysaccharides, alkaloids, etc.) and traditional Chinese medicine formulas (such as Bushen huoxue decoction, Liuwei dihuang pills, Erzhi pills, etc.) have been proven to promote bone formation, inhibit bone resorption, enhance bone cell autophagy, and delay the progression of osteoporosis by regulating the mTOR signaling pathway. Therefore, the article summarizes the traditional Chinese medicine monomer and formula that intervene in the mTOR signaling pathway for the treatment of osteoporosis, in order to provide medication ideas for the traditional Chinese medicine treatment of osteoporosis.
4.Primary central nervous system T-cell lymphoma: a case report
Bo LI ; Changguo SHAN ; Weiping HONG ; Hainan LI ; Tao LIN ; Wensheng WANG ; Zhanhang WANG ; Linbo CAI
Chinese Journal of Neurology 2022;55(12):1396-1401
Primary central nervous system T-cell lymphomas (PCNSTL) are rare, the clinical symptoms and radiographic imaging of which are unspecific, and the pathological morphology is antypical, leading to misdiagnosis and delays in treatment. A 45-year-old male patient with diplopia accompanied by numbness and dysarthria was reported in this paper, which was considered as "lymphoma or lymphoproliferative lesions" on magnetic resonance imaging (MRI) while no typical tumor cells in brain biopsy. The clinical symptoms worsened one month later and the reexamined MRI showed that the scope of the lesion was enlarged and the enhancement was more obvious than before, which was still considered as lymphoma or lymphoproliferative lesion. The second biopsy was performed and still no typical tumor lymphocytes were seen. Finally, gene rearrangement was carried out and showed the β and γ chains both present positive mutations in T cell receptor (TCR) gene rearrangement. Combined with cell morphology, immunophenotype and TCR gene rearrangement results, the patient was finally diagnosed as PCNSTL. This article reviewed the clinical symptoms, imaging features, laboratory examinations, pathological characteristics, diagnosis and differential diagnosis of PCNSTL, so as to improve the understanding of this rare disease.
5.Preliminary longitudinal mechanical pattern study of the isolated carotid atherosclerotic plaque intima using velocity vector imaging
Wensheng YUE ; Lixue YIN ; Shan WANG ; Yan DENG ; Mingliang ZUO ; Shuang LI ; Yang YU ; Anguo LUO
Chinese Journal of Ultrasonography 2008;17(8):681-684
Objective To evaluate the mechanical homogeneity pattern of isolated carotid atherosclerotic plaque intima on longitudinal axis view using ultrasonic velocity vector imaging.Methods Fourty six patients with 48 isolated plaques were undergone the high frequency ultrasound scanning,the real-time two-dimensional longitudinal-axis gray scale view at the maximal thickness of the plaque was obtained in three complete cardiac cycles.A dedicated velocity vector imaging(ⅤⅤⅠ)workstation was used for the off-line dynamic two-dimensional gray-scale image analysis and mechanics parameter(i.e.,the peak strain and the absolute difference of the peak strain)assessment at the sites of the upper,middle and down stream separately.The length and direction distribution of the velocity vectors at the carotid atherosclerotic plaque intima were observed.Wilcoxon test was utilized for the comparison of the peak strain and the absolute difference of the peak strain between two sites.Results The chaos phenomenon of velocity vector distribution at the sites of plaque intima occurred significantly differed from those uniform velocity vectors at the reference sites.According to the two groups of the higher strain value side and the lower strain value side on the upper stream and the down stream of the plaque intima,the peak strain at the higher strain value side was significantly higher than the lower strain value side(P=0.000).There was a significantly difference between the absolute difference of peak strain from the higher strain value side to the middle stream point of the plaque intima and the absolute difference of peak strain from the lower strain value side to the higher strain value side of the plaque intima(P<0.017).Conclusions The length and direction distribution of velocity vectors of the isolated carotid atherosclerotic plaque intima on longitudinal axis view are uneven,the mechanical heterogeneity pattern exists at the plaque intima and could be visualized and quantitatively evaluated using ultrasonic velocity vector imaging.

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