ABSTRACT
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.
ABSTRACT
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 × 109 l
ABSTRACT
Objective No studies have been reported on the comparison of ultracentrifugation, ExoPerfectTM-MU and PEG6000 in extracting seminal plasma exosomes. This article aimed to compare the three methods for the extraction and identification of seminal plasma exosomes. Methods Semen samples were obtained from 30 healthy donors and randomly divided into three portions, followed by extraction of exosomes from the seminal plasma by ultracentrifugation, ExoPerfectTM-MU, and 8%PEG6000, respectively. The size of the extracted exosomes was measured with the nanoparticle tracking analyzer (NTA), their morphology observed under the transmission electron microscope (TEM), and their protein biomarkers detected by Western blot. Results Significantly higher expressions of CD63 and TSG101 were found in the exosomes extracted by ultracentrifugation than in those extracted by ExoPerfectTM-MU and 8%PEG6000 (P0.05). Compared with the 8%PEG6000 group, the ultracentrifugation and ExoPerfectTM-MU groups showed significantly higher concentrations ([11.90±1.78] vs [21.20±0.98] and [19.74±1.45]×108/mL, P<0.01) and numbers of seminal plasma exosomes under TEM (4.7±1.7 vs 7.0±1.6 and 6.0±1.6, P< 0.01). Conclusion Ultracentrifugation, ExoPerfectTM-MU and 8%PEG6000 are all capable of successful extraction and identification of seminal plasma exosomes, but the former two yield more exosomes, the latter one gives a higher purity, and ExoPerfectTM-MU is simple and convenient in operation.
ABSTRACT
<p><b>OBJECTIVE</b>To extract and identify semen-derived exosome using PEG6000.</p><p><b>METHODS</b>Exosomes were extracted from semen specimens from 6 healthy volunteers with step-by-step centrifugations and ultracentrifugation prior to 8% PEG6000 enrichment. The extracted exosomes were characterized by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA) and Western blotting.</p><p><b>RESULTS</b>The pellets obtained were round or elliptic membrane vesicles 30 to 150 nm in diameter with intact double membranes and contained low electron density material. The pellets expressed CD63, ALIX and TSG101 molecules but not calnexin that was expressed in sperm cells.</p><p><b>CONCLUSION</b>The PEG6000-based method for extraction of exosomes from semen samples facilitates future studies of seminal exosomes.</p>
ABSTRACT
<p><b>Objective</b>To explore the application value of real-time contrast-enhanced ultrasound (RTCEU) in improving the detection rate of transrectal ultrasound-guided prostate biopsy.</p><p><b>METHODS</b>This prospective study included 91 male patients with abnormally high PSA (4－20 μg/L) or abnormalities in DRE or MRI, who underwent 12+X prostate biopsy following conventional transrectal ultrasonography (TRUS) and RTCEU examination. We compared the numbers of suspected prostatic nodules before and after RTCEU as well as the detection rates of prostate cancer between conventional TRUS-guided 12PBx and 12PBx plus lesion-targeted biopsy procedures.</p><p><b>RESULTS</b>Totally, 57 of the 86 suspected lesions on TRUS (66.3%), and 108 of the 118 abnormal nodules on RTCEU (91.5%) were confirmed to be prostate cancer. RTCEU achieved a significantly higher detection rate than TRUS (P<0.01). A total of 39 cases of prostate cancer (42.8%) were detected by RTCEU, while only 28 (30.7%) by TRUS, with statistically significant difference in the detection rate between the two procedures (P=0.033).</p><p><b>CONCLUSIONS</b>Real-time contrast-enhanced ultrasound can significantly improve the detection rate of prostate cancer and provide a valuable guide to targeted prostate biopsy.</p>
Subject(s)
Humans , Male , Contrast Media , Endoscopic Ultrasound-Guided Fine Needle Aspiration , Magnetic Resonance Imaging , Prospective Studies , Prostate , Diagnostic Imaging , Pathology , Prostate-Specific Antigen , Blood , Prostatic Neoplasms , Blood , Diagnostic Imaging , Pathology , Ultrasonography, InterventionalABSTRACT
<p><b>Objective</b>To investigate the changes in the percentage of myeloid-derived suppressor cells (MDSCs) in the peripheral blood of prostate cancer (PCa) patients and explore the correlation of MDSCs and their subsets with the prognosis of PCa.</p><p><b>METHODS</b>Using flow cytometry, we determined the percentage of MDSCs and the levels of Arg-1, iNOS and PD-L1 in the peripheral blood of 32 PCa patients and 25 healthy controls, detected the distribution of CD14+ Mo-MDSC and CD15+ PMN-MDSC subsets, and analyzed the correlation between the obtained parameters and the prognosis of PCa.</p><p><b>RESULTS</b>Compared with the healthy controls, the PCa patients showed significant increases in the percentage of MDSCs (P<0.01) and levels of Arg-1, iNOS and PD-L1 in the peripheral blood. Statistically significant differences were observed in the distribution of the CD14+ Mo-MDSC and CD15+ PMN-MDSC subsets between the two groups（60.4% vs 72.2%, 29.5% vs 18.8%） (P<0.05). The percentages of MDSCs and Mo-MDSCs were remarkably correlated with the total survival rate of the PCa patients (P=0.025 and 0.017).</p><p><b>CONCLUSIONS</b>The percentages of MDSCs and CD14+ Mo-MDSCs in the peripheral blood were correlated with the prognosis of PCa, which may provide a target or some evidence for the clinical treatment of PCa.</p>