Peter V. Glybochko

Endothelial to mesenchymal transition contributes to arsenic-trioxide-induced cardiac fibrosis.

Emerging evidence has suggested the critical role of endothelial to mesenchymal transition (EndMT) in fibrotic diseases. The present study was designed to examine whether EndMT is involved in arsenic trioxide (As2O3)-induced cardiac fibrosis and to explore the underlying mechanisms. Cardiac dysfunction was observed in rats after exposure to As2O3 for 15 days using echocardiography, and the deposition of collagen was detected by Masson’s trichrome staining and electron microscope. EndMT was indicated by the loss of endothelial cell markers (VE-cadherin and CD31) and the acquisition of mesenchymal cell markers (α-SMA and FSP1) determined by RT-PCR at the mRNA level and Western blot and immunofluorescence analysis at the protein level. In the in-vitro experiments, endothelial cells acquired a spindle-shaped morphology accompanying downregulation of the endothelial cell markers and upregulation of the mesenchymal cell markers when exposed to As2O3. As2O3 activated the AKT/GSK-3β/Snail signaling pathway, and blocking this pathway with PI3K inhibitor (LY294002) abolished EndMT in As2O3-treated endothelial cells. Our results highlight that As2O3 is an EndMT-promoting factor during cardiac fibrosis, suggesting that targeting EndMT is beneficial for preventing As2O3-induced cardiac toxicity.

Prognostic Value of PD-1 and PD-L1 Expression in Patients with High Grade Upper Tract Urothelial Carcinoma

Purpose: We investigated the prognostic value of PD‐1 and PD‐L1 expression in patients with high grade upper tract urothelial carcinoma. Materials and Methods: Tissue microarrays of 423 patients treated with extirpative surgery for high grade upper tract urothelial carcinoma from the International Upper Tract Urothelial Carcinoma collaboration were stained for PD‐1 and PD‐L1 using antibodies, including Cell Marque™ NAT105 diluted 1:250 and prediluted E1L3N® via immunohistochemistry. A 1% or greater staining rate of tumor infiltrating lymphocytes (PD‐1) and tumor cells (PD‐L1) was considered positive. Univariate and multivariate analyses were performed to assess independent prognosticators of survival outcomes. Results: Median patient age was 70.0 years and median followup was 37.0 months. PD‐1 and PD‐L1 were positive in 37.2% and 26.2% of patients, respectively. PD‐1 positivity was significantly associated with adverse pathological characteristics while PD‐L1 positivity was associated with favorable pT stage. On univariate analysis PD‐1 expression was associated with worse recurrence‐free, cancer specific and overall survival. On multivariate analysis PD‐1 expression was an independent prognosticator of cancer specific survival (HR 1.7, 95% CI 1.03–2.66, p = 0.039) and overall survival (HR 1.5, 95% CI 1.05–2.24, p = 0.029) but not recurrence‐free survival (HR 1.4, 95% CI 0.9–2.16, p = 0.139). On univariate analysis PD‐L1 expression was not significantly associated with survival outcomes. However, on multivariate analysis in patients with organ confined disease (pT2 or less, pN0/x and cM0), PD‐L1 positivity was an independent prognosticator of recurrence‐free survival (HR 0.2, 95% CI 0.06–0.98, p = 0.046) and overall survival (HR 0.3, 95% CI 0.11–0.63, p = 0.003). Conclusions: PD‐1 positivity of tumor‐infiltrating lymphocytes was associated with adverse pathological criteria and independent prognostication of worse survival outcomes. PD‐L1 positivity of tumor cells was an independent prognosticator of favorable survival outcomes in cases of organ confined disease.

miR-93-3p inhibition suppresses clear cell renal cell carcinoma proliferation, metastasis and invasion

miRNA dysregulation is associated with many human diseases, including cancer. This study explored the effects of miR-93-3p on clear cell renal cell carcinoma (ccRCC). We found that miR-93-3p is upregulated an average of 38-fold in 138 ccRCC specimens compared to matched normal kidney tissues, which correlated with poor patient outcome. miR-93-3p inhibition reduced ccRCC cell growth, invasion, and migration in vitro and in a mouse xenograft model. A search of the TargetScan, miRanda, and PicTar databases revealed that miR-93-3p is predicted to regulate pigment epithelium-derived factor (PEDF). A direct PEDF-miR-93-3p interaction was confirmed via dual-luciferase reporter assays. Like miR-93-3p inhibition, PEDF overexpression induced cell apoptosis and inhibited migration and invasion. Additionally, co-transfection with PEDF siRNA reversed the effects of miR-93-3p inhibition in ccRCC cells. Thus, miR-93-3p is a likely ccRCC oncogene that acts by regulating PEDF. These results suggest that miR-93-3p may predict ccRCC patient clinical outcome and serve as a novel anti-ccRCC therapeutic target.

Postoperative Nomogram for Relapse-Free Survival in Patients with High Grade Upper Tract Urothelial Carcinoma

Purpose: We developed a prognostic nomogram for patients with high grade urothelial carcinoma of the upper urinary tract after extirpative surgery. Materials and Methods: Clinical data were available for 2,926 patients diagnosed with high grade urothelial carcinoma of the upper urinary tract who underwent extirpative surgery. Cox proportional hazard regression models identified independent prognosticators of relapse in the development cohort (838). A backward step‐down selection process was applied to achieve the most informative nomogram with the least number of variables. The L2‐regularized logistic regression was applied to generate the novel nomogram. Harrell’s concordance indices were calculated to estimate the discriminative accuracy of the model. Internal validation processes were performed using bootstrapping, random sampling, tenfold cross‐validation, LOOCV, Brier score, information score and F1 score. External validation was performed on an external cohort (2,088). Decision tree analysis was used to develop a risk classification model. Kaplan‐Meier curves were applied to estimate the relapse rate for each category. Results: Overall 35.3% and 30.7% of patients experienced relapse in the development and external validation cohort. The final nomogram included age, pT stage, pN stage and architecture. It achieved a discriminative accuracy of 0.71 and 0.76, and the AUC was 0.78 and 0.77 in the development and external validation cohort, respectively. Rigorous testing showed constant results. The 5‐year relapse‐free survival rates were 88.6%, 68.1%, 40.2% and 12.5% for the patients with low risk, intermediate risk, high risk and very high risk disease, respectively. Conclusions: The current nomogram, consisting of only 4 variables, shows high prognostic accuracy and risk stratification for patients with high grade urothelial carcinoma of the upper urinary tract following extirpative surgery, thereby adding meaningful information for clinical decision making.

A Feasibility Study Utilizing the Thulium and Holmium Laser in Patients for the Treatment of Recurrent Benign Prostatic Hyperplasia after Previous Prostatic Surgery

Background: Transurethral resection of the prostate (TURP) is considered to be the standard treatment for patients with benign prostatic obstruction (BPO) ≤80 mL. However, up to 14.7% of the patients require secondary TURP due to recurrent BPO. The aim of our study was to describe specific features of holmium laser enucleation of the prostate (HoLEP) and thulium laser enucleation of the prostate (ThuLEP) in patients with recurrent BPO after previous prostate surgery. Materials and Methods: A total of 768 consecutive patients from our prospective collected database were retrospectively reviewed and divided into 4 groups: group A (489 patients) and group C (253 patients) underwent primary HoLEP and ThuLEP treatment, while group B (17 patients) and D (9 patients) included patients with recurrent BPO who were treated with HoLEP and ThuLEP, respectively. Results: There were no significant differences in preoperative parameters between the groups at primary (A and C) and secondary (B and D) treatment except their age. At 6-month follow-up, voiding parameters and symptom scores showed statistically significant improvements compared to baseline without differences between the groups. The mean operative time was comparable between the groups and did not differ significantly (p > 0.05). Conclusions: Laser enucleation for the treatment of recurrent BPO is feasible and seems to be a safe and effective procedure.

Risk factors for urethral stricture and/or bladder neck contracture after monopolar transurethral resection of the prostate for benign prostatic hyperplasia:

Objective: To identify risk factors for urethral stricture and/or bladder neck contracture after transurethral resection of benign prostatic hyperplasia. Materials and methods: We performed a retrospective analysis of 402 patients, which underwent a monopolar transurethral resection of the prostate in the urology clinic of Sechenov First Moscow State Medical University for prostatic hyperplasia during the period 2011–2014. Urethral stricture and (or) bladder neck contracture in the postoperative period were diagnosed in 61 (15.27%) patients; 34 patients (8.6%) had urethral stricture, 20 (4.97%) bladder neck contracture, and 7 (1.7%) had a combination of urethral stricture and bladder neck contracture. In 341 of cases (84.73%), no late postoperative complications were observed. A total of 106 of the 341 patients met the inclusion criteria, hence, containing all the information necessary for analysis such as the volume of the prostate, the duration of the surgery, the size of the endoscope, data on concomitant diseases, analysis prostatic secretion, and so on. Thus, two groups were formed. Group 1 (106 patients) is the control group in which urethral strictures and/or bladder neck contractures did not occur in the long-term postoperative period and group 2 (61 patients), in which was observed the formation of these complications. To calculate the statistical significance of the differences for categorical data, Fisher criterion was used. For quantitative variables, in the case of normal data distribution, an unpaired t-test or one-way analysis of variance was used; for data having a distribution different from normal, a Mann–Whitney rank test was used. Results: Regression analysis established the significance of the influence of four factors on the development of scar-sclerotic changes of urethra and bladder neck: the tool diameter 27 Fr (p < 0.0001), presence of prostatitis in past medical history (p < 0.0001), prostate volume (p = 0.003), and redraining of the bladder (p = 0.0162). Conclusion: The relationship between the diameter of the instrument, presence of chronic prostatitis in anamnesis, increased volume of the prostate, and repeated drainage of the bladder using the urethral catheter with the risk of developing scar-sclerotic changes in the urethra and/or bladder neck are statistically reliable and confirmed as a result of regression analysis.

Multiple application of three-dimensional soft kidney models with localized kidney cancer: A pilot study:

Aim: To evaluate the effectiveness of three-dimensional printing application in urology for localized renal cancer treatment using three-dimensional printed soft models. Materials and methods: The study included five patients with kidney tumors. The patients were treated in the Urology Clinic of I.M. Sechenov First Moscow State Medical University from February 2016 to June 2017. Personalized three-dimensional printed models based on computed tomographic images were created. Five surgeons took part in a survey in which the utility of computed tomographic images versus three-dimensional printed models for presurgical planning was compared. A laparoscopic partial nephrectomy training using the developed three-dimensional printed models was performed by the same surgeons in a surgical training box. Results: The patients underwent endoscopic surgery using laparoscopic access. The average time of surgery was 187 min. All the operations were performed with complete renal artery clamping. The average warm ischemia time was 19.5 min and the average blood loss was 170 mL. No conversions to open surgery or radical nephrectomy, and no postoperative complications and deaths were observed. All the surgical margins were negative. The tumors were morphologically identified as renal cell carcinoma in four cases and as oncocytoma in one case. Conclusion: The developed three-dimensional printed models allow one to evaluate the pathological anatomy of tumors more effectively. High similarity between three-dimensional models and native kidneys contribute to improvement of surgical skills necessary for partial nephrectomy. Training on the three-dimensional models also allows surgeons to facilitate selection of an optimal surgical tactics for each patient.

The clinical role of X-ray computed tomography to predict the clinical efficiency of extracorporeal shock wave lithotripsy

Aim: To evaluate the clinical efficiency of computed tomography for diagnostics of patients with urolithiasis and the choice of treatment strategy. Material and methods: The study was carried out at the Urological Clinic of I.M. Sechenov First Moscow State Medical University and included 1044 patients with urinary stones. The ultimate goal of this study was to predict the clinical efficiency of extracorporeal shock wave lithotripsy using a combination of computed tomography and densitometry. Extracorporeal shock wave lithotripsy was performed on “Siemens Lithostar Plus,” “Siemens Modularis Uro,” and “Dornier Gemini” lithotripters. Statistical analysis of clinical data included evaluation of individual sampling groups and calculation of weighted arithmetic mean (M). Results: The efficiency of extracorporeal shock wave lithotripsy has been determined primarily using X-ray analysis of the concrement outlines and the structure (homogeneous or heterogeneous) of its central zone. However, in terms of efficiency and repetition rate (the number of fragmentation procedures required for complete clearance) of extracorporeal shock wave lithotripsy, the mean density of the concrement along the whole length of its three-dimensional structure (expressed in Hounsfield units) appeared to be the most reliable and informative predictive index in this study. Conclusion: The combination of computed tomography with densitometry in the treatment of patients with urolithiasis allows one (1) to determine the exact localization, size, X-ray structure, and structural density of urinary stones and (2) to predict, on the basis of densitometric data histograms, the clinical efficiency and repetition rate of extracorporeal shock wave lithotripsy with due regard to the X-ray structure of peripheral and central zones, and mean density (in Hounsfield units) of urinary concrements.

Identification and Functional Verification of MicroRNAs in the Obese Rat With Erectile Dysfunction

Introduction Obesity is a potential risk factor for erectile dysfunction (ED). MicroRNAs (miRNAs) regulate the expression of genes involved in various pathophysiologic processes. Aim To identify the miRNA profile in the corpus cavernosum (CC) of obese rats with ED and elucidate the potential function of miRNA in the pathogenesis of ED. Methods Obesity was induced in rats by a high-fat diet. After the erectile function test, experimental animals were divided into two groups: obese rats with ED and obese rats with normal erectile function. The CCs from these rats were collected for miRNA microarray analysis. The results were verified by real-time polymerase chain reaction analysis. Subsequently, the targets of differentially expressed miRNAs were predicted. Bioinformatics analysis was applied to predict the functions of differentially expressed miRNAs in ED. Apomorphine-induced penile erection and intracavernous pressure measurements were used to evaluate the effects of miRNA on the erectile function of rats. Main Outcome Measures MiRNA expression in the CC of obese rats with ED and those with normal erectile function was detected by miRNA microarray analysis. Candidate miRNAs were validated by real-time polymerase chain reaction. Bioinformatics analysis was used to predict the functions of miRNAs. Apomorphine-induced penile erection and intracavernous pressure measurements were used to reflect the erectile function of rats. Results Sixty-eight miRNAs were differentially expressed in the CC of obese rats with ED (≥1.5-fold change). The real-time polymerase chain reaction results were consistent with the miRNA microarray analysis results. Specifically, miR-328a was significantly upregulated in rats with ED compared with control rats and was chosen for functional evaluation in the pathogenesis of ED. Overexpression of miR-328a noticeably decreased the erectile response to apomorphine and the expression of heme oxygenase-1. Conclusion MiRNAs are involved in the pathogenesis of obesity-related ED. MiR-328a might facilitate the induction of ED. Bai Y, Zhang L, Jiang Y, et al. Identification and Functional Verification of MicroRNAs in the Obese Rat With Erectile Dysfunction. Sex Med 2017;5:e261–e271.

The Role of Water-Jet Dissection in Improving Erectile Function and Urinary Continence after Nerve-Sparing Prostatectomy

Objectives The aim of this study was to assess erectile function (EF) and urinary continence (UC) recovery after nerve-sparing retropubic radical prostatectomy (NS-RRPE) and nerve-sparing laparoscopic radical prostatectomy (NS-LRPE) depending on the neurovascular bundle (NVB) dissection technique used. Methods Twenty-four hour pad test was used to assess the degree of postoperative UC 1, 6 and 12 months after the surgery. For the purpose of EF assessment, International Index of Erectile Function-5 (IIEF-5) questionnaire was used. In addition, EF recovery assessment included the time needed to achieve a successful sexual intercourse with or without Inhibitors Phosphodiesterase 5 (IPDE5). A successful sexual intercourse was defined as an ability to achieve a strong erection enough for penetration and maintain erection for a long time, throughout the intercourse. Assessment was performed before the surgery and 8 and 6 months after the surgery. Results In the patient group with BNS + water jet dissection (WJD), the IIEF-5 score was considerably higher 8 weeks after the surgery (by 2.8 points) (р = 0.02). In 6 months, the difference between the group become more significant and reached 3.5 points (p = 0.01). Three months after urethral catheter removal, majority (95%) of patients in the NS-RPE + WJD group had no urinary incontinence (UI). For standard NS-RPE, that figure was 87%. Mild stress UI was observed in 5% of patients after NS-RPE + WJD and in 13% of patients after NS-RPE. Six months after urethral catheter removal, no significant differences in UC assessment were observed in the two groups. Conclusions The implementation of WJD of NVB in clinical practice has made it possible to considerably improve the quality of life for postoperative patients due to good outcomes in terms of early UC and EF recovery.