Cyclophosphamide-induced testosterone deficiency (CPTD) during the treatment of cancers and autoimmune disorders severelyinfluences the quality of life of patients. Currently, several guidelines recommend patients suf...Cyclophosphamide-induced testosterone deficiency (CPTD) during the treatment of cancers and autoimmune disorders severelyinfluences the quality of life of patients. Currently, several guidelines recommend patients suffering from CPTD receive testosteronereplacement therapy (TRT). However, TRT has many disadvantages underscoring the requirement for alternative, nontoxictreatment strategies. We previously reported bone marrow mesenchymal stem cells-derived exosomes (BMSCs-exos) could alleviatecyclophosphamide (CP)-induced spermatogenesis dysfunction, highlighting their role in the treatment of male reproductive disorders.Therefore, we further investigated whether BMSCs-exos affect autophagy and testosterone synthesis in Leydig cells (LCs). Here,we examined the effects and probed the molecular mechanisms of BMSCs-exos on CPTD in vivo and in vitro by detecting theexpression levels of genes and proteins related to autophagy and testosterone synthesis. Furthermore, the testosterone concentrationin serum and cell-conditioned medium, and the photophosphorylation protein levels of adenosine monophosphate-activatedprotein kinase (AMPK) and mammalian target of rapamycin (mTOR) were measured. Our results suggest that BMSCs-exos couldbe absorbed by LCs through the blood–testis barrier in mice, promoting autophagy in LCs and improving the CP-induced low serumtestosterone levels. BMSCs-exos inhibited cell death in CP-exposed LCs, regulated the AMPK-mTOR signaling pathway to promoteautophagy in LCs, and then improved the low testosterone synthesis ability of CP-induced LCs. Moreover, the autophagy inhibitor,3-methyladenine (3-MA), significantly reversed the therapeutic effects of BMSCs-exos. These findings suggest that BMSCs-exospromote LC autophagy by regulating the AMPK-mTOR signaling pathway, thereby ameliorating CPTD. This study provides novelevidence for the clinical improvement of CPTD using BMSCs-exos.展开更多
Accurate methods for identifying pelvic lymph node metastasis(LNM)of prostate cancer(PCa)prior to surgery are still lacking.We aimed to investigate the predictive value of peripheral monocyte count(PMC)for LNM of PCa ...Accurate methods for identifying pelvic lymph node metastasis(LNM)of prostate cancer(PCa)prior to surgery are still lacking.We aimed to investigate the predictive value of peripheral monocyte count(PMC)for LNM of PCa in this study.Two hundred and ninety-eight patients from three centers were divided into a training set(n=125)and a validation set(n=173).In the training set,the independent predictors of LNM were analyzed using univariate and multivariate logistic regression analyses,and the optimal cutoff value was calculated by the receiver operating characteristic(ROC)curve.The sensitivity and specificity of the optimal cutoff were authenticated in the validation cohort.Finally,a nomogram based on the PMC was constructed for predicting LNM.Multivariate analyses of the training cohort demonstrated that clinical T stage,preoperative Gleason score,and PMC were independent risk factors for LNM.The subsequent ROC analysis showed that the optimal cutoff value of PMC for diagnosing LNM was 0.405×10^(9)l^(−1)with a sensitivity of 60.0%and a specificity of 67.8%.In the validation set,the optimal cutoff value showed significantly higher sensitivity than that of conventional magnetic resonance imaging(MRI)(0.619 vs 0.238,P<0.001).The nomogram involving PMC,free prostate-specific antigen(fPSA),clinical T stage,preoperative Gleason score,and monocyte-to-lymphocyte ratio(MLR)was generated,which showed a robust predictive capacity for predicting LNM before the operation.Our results indicated that PMC as a single agent,or combined with other clinical parameters,showed a robust predictive capacity for LNM in PCa.It can be employed as a complementary factor for the decision of whether to conduct pelvic lymph node dissection.展开更多
Spermatogenic dysfunction caused by cyclophosphamide(CP)chemotherapy has seriously influenced the life quality of patients.Unfortunately,treatments for CP-induced testicular spermatogenic dysfunction are limited,and t...Spermatogenic dysfunction caused by cyclophosphamide(CP)chemotherapy has seriously influenced the life quality of patients.Unfortunately,treatments for CP-induced testicular spermatogenic dysfunction are limited,and the molecular mechanisms are not fully understood.For the first time,here,we explored the effects of bone marrow mesenchymal stem cell-derived exosomes(BMSC-exos)on CP-induced testicular spermatogenic dysfunction in vitro and in vivo.BMSC-exos could be taken up by spermatogonia(GC1-spg cells).CP-injured GC1-spg cells and BMSC-exos were cocultured at various doses,and then,cell proliferation was measured using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide(MTT)assay.In addition,photophosphorylation of extracellular-regulated kinase(ERK),p38 mitogen-activated protein kinase(p38MAPK),and protein kinase B(AKT)proteins was evaluated by western blotting as well as apoptosis in GC1-spg cells measured using flow cytometry.Treatment with BMSC-exos enhanced cell proliferation and reduced apoptosis of CP-injured GCI-spg cells.Phosphorylated levels of ERK,AKT,and p38MAPK proteins were reduced in CP-injured spermatogonia when co-treated with BMSC-exos,indicating that BMSC-exos acted against the reproductive toxicity of CP via the p38MAPK/ERK and AKT signaling pathways.In experiments in vivo,CP-treated rats received BMSC-exos by injection into the tail vein,and testis morphology was compared between treated and control groups.Histology showed that transfusion of BMSC-exos inhibited the pathological changes in CP-injured testes.Thus,BMSC-exos could counteract the reproductive toxicity of CP via the p38MAPK/ERK and AKT signaling pathways.The findings provide a potential treatment for CP-induced male spermatogenic dysfunction using BMSC-exos.展开更多
基金supported by grants from the National Natural Science Foundation of China(No.81772257)the Third Affiliated Hospital of Southern Medical University President’s Fund(YM2021008).
文摘Cyclophosphamide-induced testosterone deficiency (CPTD) during the treatment of cancers and autoimmune disorders severelyinfluences the quality of life of patients. Currently, several guidelines recommend patients suffering from CPTD receive testosteronereplacement therapy (TRT). However, TRT has many disadvantages underscoring the requirement for alternative, nontoxictreatment strategies. We previously reported bone marrow mesenchymal stem cells-derived exosomes (BMSCs-exos) could alleviatecyclophosphamide (CP)-induced spermatogenesis dysfunction, highlighting their role in the treatment of male reproductive disorders.Therefore, we further investigated whether BMSCs-exos affect autophagy and testosterone synthesis in Leydig cells (LCs). Here,we examined the effects and probed the molecular mechanisms of BMSCs-exos on CPTD in vivo and in vitro by detecting theexpression levels of genes and proteins related to autophagy and testosterone synthesis. Furthermore, the testosterone concentrationin serum and cell-conditioned medium, and the photophosphorylation protein levels of adenosine monophosphate-activatedprotein kinase (AMPK) and mammalian target of rapamycin (mTOR) were measured. Our results suggest that BMSCs-exos couldbe absorbed by LCs through the blood–testis barrier in mice, promoting autophagy in LCs and improving the CP-induced low serumtestosterone levels. BMSCs-exos inhibited cell death in CP-exposed LCs, regulated the AMPK-mTOR signaling pathway to promoteautophagy in LCs, and then improved the low testosterone synthesis ability of CP-induced LCs. Moreover, the autophagy inhibitor,3-methyladenine (3-MA), significantly reversed the therapeutic effects of BMSCs-exos. These findings suggest that BMSCs-exospromote LC autophagy by regulating the AMPK-mTOR signaling pathway, thereby ameliorating CPTD. This study provides novelevidence for the clinical improvement of CPTD using BMSCs-exos.
基金This study was supported by the National Natural Science Foundation of China(NSFC 81602248)the Natural Science Foundation of Guangdong Province(No.2017A030313686)to MKC.
文摘Accurate methods for identifying pelvic lymph node metastasis(LNM)of prostate cancer(PCa)prior to surgery are still lacking.We aimed to investigate the predictive value of peripheral monocyte count(PMC)for LNM of PCa in this study.Two hundred and ninety-eight patients from three centers were divided into a training set(n=125)and a validation set(n=173).In the training set,the independent predictors of LNM were analyzed using univariate and multivariate logistic regression analyses,and the optimal cutoff value was calculated by the receiver operating characteristic(ROC)curve.The sensitivity and specificity of the optimal cutoff were authenticated in the validation cohort.Finally,a nomogram based on the PMC was constructed for predicting LNM.Multivariate analyses of the training cohort demonstrated that clinical T stage,preoperative Gleason score,and PMC were independent risk factors for LNM.The subsequent ROC analysis showed that the optimal cutoff value of PMC for diagnosing LNM was 0.405×10^(9)l^(−1)with a sensitivity of 60.0%and a specificity of 67.8%.In the validation set,the optimal cutoff value showed significantly higher sensitivity than that of conventional magnetic resonance imaging(MRI)(0.619 vs 0.238,P<0.001).The nomogram involving PMC,free prostate-specific antigen(fPSA),clinical T stage,preoperative Gleason score,and monocyte-to-lymphocyte ratio(MLR)was generated,which showed a robust predictive capacity for predicting LNM before the operation.Our results indicated that PMC as a single agent,or combined with other clinical parameters,showed a robust predictive capacity for LNM in PCa.It can be employed as a complementary factor for the decision of whether to conduct pelvic lymph node dissection.
基金This study was supported by grants from the National Natural Science Foundation of China(No.81772257)the Guangdong Provincial Natural Science Foundation of China(No.2018A030313697).
文摘Spermatogenic dysfunction caused by cyclophosphamide(CP)chemotherapy has seriously influenced the life quality of patients.Unfortunately,treatments for CP-induced testicular spermatogenic dysfunction are limited,and the molecular mechanisms are not fully understood.For the first time,here,we explored the effects of bone marrow mesenchymal stem cell-derived exosomes(BMSC-exos)on CP-induced testicular spermatogenic dysfunction in vitro and in vivo.BMSC-exos could be taken up by spermatogonia(GC1-spg cells).CP-injured GC1-spg cells and BMSC-exos were cocultured at various doses,and then,cell proliferation was measured using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide(MTT)assay.In addition,photophosphorylation of extracellular-regulated kinase(ERK),p38 mitogen-activated protein kinase(p38MAPK),and protein kinase B(AKT)proteins was evaluated by western blotting as well as apoptosis in GC1-spg cells measured using flow cytometry.Treatment with BMSC-exos enhanced cell proliferation and reduced apoptosis of CP-injured GCI-spg cells.Phosphorylated levels of ERK,AKT,and p38MAPK proteins were reduced in CP-injured spermatogonia when co-treated with BMSC-exos,indicating that BMSC-exos acted against the reproductive toxicity of CP via the p38MAPK/ERK and AKT signaling pathways.In experiments in vivo,CP-treated rats received BMSC-exos by injection into the tail vein,and testis morphology was compared between treated and control groups.Histology showed that transfusion of BMSC-exos inhibited the pathological changes in CP-injured testes.Thus,BMSC-exos could counteract the reproductive toxicity of CP via the p38MAPK/ERK and AKT signaling pathways.The findings provide a potential treatment for CP-induced male spermatogenic dysfunction using BMSC-exos.