ADSCs labeled with SPIONs tracked in corpus cavernosum of rat and miniature pig by MR imaging and histological examination.

Adipose tissue-derived stem cells (ADSCs) have been shown to improve erectile function in animal models of erectile dysfunction. However, few studies have been carried out using a reliable in vivo imaging method to trace transplanted cells in real time, which is necessary for systematic investigation of cell therapy. The study aims to explore the feasibility of non-invasively monitoring intracavernous injection of ADSCs in rat and miniature pig corpus cavernosum using in vivo magnetic resonance (MR) imaging. Thirty-six male Sprague Dawley rats (10 weeks old) and six healthy, sexually mature male miniature pigs (20 kg weight) were obtained. ADSCs were isolated from paratesticular fat of donor rats and cultured. Then ADSCs were labeled with superparamagnetic iron oxide nanoparticles (SPIONs), a type of MR imaging contrast agent, before transplantation into rats and pigs. After intracavernous injection, all rats and pigs underwent and were analyzed by MR imaging at the day of ADSC transplantation and follow-up at 1, 2 and 4 weeks after transplantation. In addition, penile histological examination was performed on all rats and pigs before (n = 6) and at 1 day (n = 6), 1 week (n = 6), 2 weeks (n = 6) or 4 weeks (n = 12) after ADSC transplantation. SPION-labeled ADSCs demonstrated a strong decreased signal intensity compared with distilled water, unlabeled ADSCs or agarose gel. SPION-labeled ADSCs showed a hypointense signal at all concentrations, and the greatest hypointense signal was observed at the concentration of 1 × 106. MR images of the corpus cavernosum showed a hypointense signal located at the injection site. T2*-weighted signal intensity increased over the course of 1 week after ADSCs transplantation, and demonstrated a similar MR signal with that before ADSCs transplantation. After SPION-labeled ADSC injection, T2*-weighted MR imaging clearly demonstrated a marked hypointense signal in pig corpus cavernosum. The T2*-weighted signal faded over time, similar to the MR imaging results in rats. Obvious acute inflammatory exudation was induced by intracavernous injection, and the T2*-weighted signal intensity of these exudation was higher than that of the injection site. The presence of iron was detected by Prussian blue staining, which demonstrated ADSC retention in rat corpus cavernosum. Lack of cellular infiltrations were demonstrated by H&E staining before and 4 weeks after transplantation, which indicated no negative immune response by rats. Prussian blue staining was positive for iron oxide nanoparticles at 2 weeks after transplantation. SPION-labeled ADSCs showed a clear hypointense signal on T2-weight MRI in vitro and in vivo. The MR signal intensity in the corpus cavernosum of the rats and miniature pigs faded and disappeared over time after ADSC transplantation. These findings suggested that MR imaging could trace transplanted ADSCs in the short term in the corpus cavernosum of animals.

Single-cell sequencing provides insights into the landscape of ovary in PCOS and alterations induced by CUMS.

Polycystic ovary syndrome (PCOS) is a common endocrine disorder related to psychological distress. However, the mechanism underlying increased prevalence of depression in PCOS remained unclear. This study aimed to explore the unique transcriptional landscape of ovary and offered a platform to explore the mechanism of PCOS, as well as the influences caused by depression. The PCOS rat model was established by letrozole whereas PCOS rat model with depression was established by letrozole combined with chronic unpredicted mild stress (CUMS). Then single-cell RNA sequencing (scRNA-Seq) was applied to analyze the transcriptional features of rat ovaries. Granulosa cells (GCs) and fibroblasts (Fibros) accounted for the top two clusters of total 12 cell types. There were nine clusters in GCs, related to inflammatory response, endoplasmic reticulum (ER) stress, and steroidogenesis. The expression of differentially expressed genes (DEG) Hes1 was higher in PCOS and PCOS + CUMS groups, exhibiting enhanced expression by pseudotime and positively related to inflammation. Pseudotemporal analysis revealed that inflammation contributed to the different GCs distributions. Moreover, analysis of DEGs and gene ontology (GO) function enrichment revealed CUMS aggravated inflammation in PCOS GCs possibly via interferon signaling pathway. In theca cells (TCs), nine clusters were observed and some of them were relevant to inflammation, ER stress, and lipid metabolism. DEGs Ass1, Insl3, and Ifi27 were positively related to Cyp17a1, and Ces1d might contribute to the different trajectory of TCs. Subsequent scRNA-seq revealed a signature profile of endothelial cells (ECs) and Fibros, which suggest that inflammation-induced damage of ECs and Fibro, further exacerbated by CUMS. Finally, analysis of T cells and mononuclear phagocytes (MPs) revealed the existence of immune dysfunction, among which interferon signaling played a critical role. These findings provided more knowledge for a better understanding PCOS from the view of inflammation and identified new biomarkers and targets for the treatment of PCOS with psychological diseases.NEW & NOTEWORTHY In this study, we mapped the landscape of polycystic ovary syndrome (PCOS) ovary with rat model induced by letrozole and provided a novel insight into the molecular mechanism of PCOS accompanied by chronic unpredicted mild stress (CUMS) at single-cell transcriptomic level. These observations highlight the importance of inflammation in the pathogenesis of PCOS, which might also be the bridge between PCOS and psychological diseases.

Antidepressant-like effect of acute dose of Naringin involves suppression of NR1 and activation of protein kinase A/cyclic adenosine monophosphate response element-binding protein/brain-derived neurotrophic factor signaling in hippocampus.

Naringin (Nr) has been identified to have antidepressant-like effects through repeated treatment. However, the underlying mechanism of the rapid antidepressant-like effects of Nr was still unclear. The present study used behavioral tests, classic depressive model and pharmacological methods to reveal the rapid antidepressant-like potential of Nr. We found that a single dose of Nr (20 mg/kg) produced antidepressant-like action after 2 h in the tail suspension test (TST) and forced swimming test (FST). Moreover, ketamine-like effects were also demonstrated by using the chronic mild stress model (CMS) and learned helplessness (LH), and the results showed that Nr reversed all behavioral defects, TST, FST, source preference test (SPT) in CMS, and LH testing, TST, FST in LH model, at 2 h after a single administration. In addition, Nr (20 mg/kg) could improve the abnormal expressions of NMDA receptor NR1 and PKA/CREB/BDNF pathway in hippocampus 2 h after a single administration in CMS mice. Further investigation revealed that activation of NMDA receptors by NMDA (750 mg/kg) could block the antidepressant effects of acute administration of Nr (20 mg/kg). However, the inhibition of NMDA receptors by MK-801 (0.05 mg/kg) promoted the subdose of Nr (10 mg/kg) to have antidepressant effect, which was similar to the effective dose Nr (20 mg/kg). Taken together, acute dose of Nr produces rapid antidepressant-like action, and the underlying mechanism could be through inhibiting NMDA receptors in the hippocampus.

Traditional Chinese medicine CFF-1 exerts a potent anti-tumor immunity to hinder tumor growth and metastasis in prostate cancer through EGFR/JAK1/STAT3 pathway to inhibit PD-1/PD-L1 checkpoint signaling.

BACKGROUND: Traditional Chinese Medicine (TCM) CFF-1 has been used in clinic for prostate cancer therapy in China. We reported before CFF-1 induced cell apoptosis via suppressing EGFR-related pathways, reminding us its potential role associated with antitumor immunity. PURPOSE: The study was aimed to investigate the regulatory mechanism of CFF-1 on PD-L1/PD-1-mediated tumor immune escape. METHODS: Prostate-specific antigen (PSA) test and the functional assessment of cancer therapy-prostate (FACT-P) and karnosky performance status (KPS) questionnaires were carried out to evaluate patient' condition before and after therapy. Flow cytometry (FCM) was used for analyzing cell apoptosis, T lymphocyte subsets and cell cycle. Western blotting and Immunohistochemistry (IHC) were performed to measure protein expressions. The synergy of drug combination was assessed by calculating combination index (CI). RESULTS: CFF-1 obviously decreased PSA and improved the quality of life in patients with advanced prostate cancer. PD-L1 was highly expressed in prostate cancer cells including LNCaP, 22Rv1, PC-3, DU145 and RM-1. PD-1/PD-L1 was upregulated in tumorigenesis and tumor progression of subcutaneous homograft mouse model with immune response, where CD3+ T cell subsets were declined. CFF-1 inhibited PD-L1 expression in prostate cancer cells in a time/dose-dependent manner and blocked tumor growth by suppressing PD-1/PD-L1 upregulation to promote the recovery of CD3+ T lymphocytes, especially CD4+ T cell subset, accompanied by the downregulation of CD4+ FOXP3+ T cell subset. CFF-1 also prolonged the survival and inhibited lung metastasis in tail vein prostate cancer mouse model while repressing PD-1/PD-L1. CFF-1 in combination with docetaxol (DTX) produced a synergistic effects by sensitizing the inhibitory effect of DTX on JAK1/STAT3 pathway targeting PD-L1 blockade. CONCLUSION: CFF-1 inhibited tumor growth and lung metastasis by blocking PD-1/PD-L1 to ameliorate T lymphocyte immune response through EGFR/JAK1/STAT3 pathway, suggesting that CFF-1 might be a promising treatment to resist tumor immunosuppression for prostate cancer patients.

NCAPD3 promotes prostate cancer progression by up-regulating EZH2 and MALAT1 through STAT3 and E2F1.

NCAPD3 is one of the non-SMC regulatory subunits of Condensin II, which is mainly responsible for the condensation and segregation of chromosomes during mitosis. However, its role in cancer especially in prostate cancer (PCa) and the molecular mechanism have not been clearly elucidated. Here, we find that NCAPD3 is high-expression and up-regulates the levels of EZH2 and MALAT1 in PCa. In detail, high expression of NCAPD3 increases the levels of transcription factor STAT3 and E2F1 and recruits more STAT3 and E2F1 to the promoter of EZH2 gene and more STAT3 to the promoter of MALAT1 gene, and then results in the increasing expression of both EZH2 and MALAT1 in PCa cells. In vitro and in vivo functional characterization reveals that overexpression of NCAPD3 enhances the growth of PCa cells, while knockdown of NCAPD3 impairs the growth of PCa cells. Together, our data demonstrate that NCAPD3 is a tumor-promoting factor which enhances the progression of PCa by up-regulating EZH2 and MALAT1.

A novel 6-gene signature derived from tumor-infiltrating T cells and neutrophils predicts survival of bladder urothelial carcinoma.

Intratumoral immune cells were reported to be associated with prognosis of bladder urothelial carcinoma (BUC). However, the role of immune cells related genes in BUC prognosis is less well defined. In the study, we analyzed data retrieved from the Cancer Genome Atlas database and found higher neutrophils and lower T cells infiltration in BUC tumor tissues were significantly correlated with patients' worse prognosis. Additionally, the expression levels of 164 genes were significantly correlated with T cells and neutrophils proportions. A Cox proportional-hazards model integrating 6 genes expression (EMP1, RASGRP4, HSPA1L, AHNAK, SLC1A6, and PRSS8) was identified. The 6-gene signature outperformed other clinical factors in risk prediction and was an independent prognostic factor for BUC. The findings were further conformed in three Gene Expression Omnibus datasets (n=331) and Jiangsu Province Hospital cohort (n = 46). Gene set enrichment analysis revealed that the model was highly involved in some immune-related pathways. A comprehensive nomogram combining the model and other clinical parameters was finally constructed to facilitate clinical application. In conclusion, a T cell and neutrophil-associated 6-gene prognostic model was identified for the survival prediction of BUC patients.

Efficacy and safety of single port robotic radical prostatectomy and multiport robotic radical prostatectomy: a systematic review and meta-analysis.

BACKGROUND: The purpose of this study is to compare the clinical efficacy and safety of single port (SP) robot radical prostatectomy and multiport (MP) robot radical prostatectomy. METHODS: Using the China National Knowledge database, EMBASE, Cochrane library, PubMed, and other databases to obtain relevant research, SP robot radical prostatectomy and MP robot radical prostatectomy were comprehensively evaluated. The software used to evaluate the impact of the results in the selected articles was Review Manager 5.2. Deviation analysis, forest plot analysis, and sensitivity analysis were carried out for the collected data. RESULTS: A total of 7 related studies that met the criteria were finally included. The data showed that the operation time of MP in the control group was significantly longer than that in the SP group [mean difference (MD) =-13.29; 95% confidence interval (CI): (-17.35, -9.23); P<0.00001; I2=50%]. The duration of intensive care unit (ICU) stay for SP surgery was shorter than that for MP surgery [MD =-18.30; 95% CI: (-29.17, -7.42); P=0.0010; I2=94%]. The blood loss of SP surgery was less than that of MP surgery [MD =-15.54; 95% CI: (-28.37, -2.71); the total effective rate was 0.02; I2=0%]. There was no significant difference in the incidence of postoperative complications between SP and MP surgery [risk ratio (RR) =0.95; 95% CI: (0.55, 1.63); P=0.85; I2=0%]. At the same time, the sensitivity analysis and funnel plot showed that this study was robust and publication bias was limited. DISCUSSION: Our results show that SP robotic radical prostatectomy is superior to MP robotic radical prostatectomy in terms of efficacy and safety. SP robot radical prostatectomy is worthy of wide promotion.

Comparison of Laparoendoscopic Single-Site and Laparoscopic Radical Cystectomy for Male Patients with Bladder Cancer: Perioperative and Oncologic Outcomes.

Objective: To compare the surgical and early oncological outcomes in patients with bladder cancer who had laparoendoscopic single-site radical cystectomy (RC) or laparoscopic RC. Materials and Methods: From July 2012 to May 2019, 28 consecutive men suffering from bladder cancer underwent laparoendoscopic single-site RC or laparoscopic RC with extracorporeally ileal conduit diversion. Data regarding the patient characteristics, surgical outcomes, and short-term oncological outcomes were analyzed retrospectively. Results: Compared with laparoscopic RC, laparoendoscopic single-site RC was associated with less postoperative pain (mean, 4.67 versus 6.08 scores; P = .004), and shorter convalescence (time to ambulation, mean, 1.13 days versus 2.15 days; P = .000; hospital stay after surgery, mean, 13 days versus 19 days; P = .001). In addition, differences in patient characteristics, mean total operation time, and mean estimated blood loss were not statistically significant between laparoendoscopic single-site RC and laparoscopic RC groups. There was no difference in the early or late complication rate between the two groups as well. It is also revealed that there was no significant difference in the overall survival rate at 24 months between laparoendoscopic single-site RC and laparoscopic RC groups. Conclusions: Based on our initial experience with laparoendoscopic single-site RC, it is a safe procedure with acceptable complications and oncological outcomes. Notably, laparoendoscopic single-site RC is associated with less postoperative pain and rapider convalescence compared with the historical series of laparoscopic RC. However, further comparative studies with longer follow-up period are warranted to validate this procedure.

Inhibition of Specificity Protein 1 Is Involved in Phloretin-Induced Suppression of Prostate Cancer.

Phloretin is a flavonoid existed in various plants and has been reported to possess anticarcinogenic activity. However, the anticancer mechanism of phloretin in prostate cancer (PCa) remains unclear. Here, our in vitro and in vivo experimental data demonstrate that phloretin inhibits the phosphorylation and the activation of EGFR and then inhibits its downstream PI3K/AKT and MEK/ERK1/2 pathways in PCa cells. Inhibition of these two pathways further decreases expression of Sp1 by inhibiting Sp1 gene transcription, induces degradation of Sp1 protein by inhibiting GSK3ß phosphorylation, suppresses nucleolin-enhanced translation of Sp1 mRNA by inhibiting nucleolin phosphorylation, and directly inactivates transcription activity of Sp1. Inhibition of Sp1 subsequently decreases the expression of Sp3/4, VEGF, and Survivin and then upregulates apoptosis-related proteins and downregulates cell cycle-related proteins in PCa cells. Finally, phloretin treatment in PCa cells induces cell growth inhibition and apoptosis, suggesting that phloretin may be an effective therapy compound in the treatment of prostate cancer.