PRMT5-activated c-Myc promote bladder cancer proliferation and invasion through up-regulating NF-κB pathway.

AIM: PRMT5 and c-Myc were considered as oncogene of bladder cancer. Nevertheless, whether the interaction between of PRMT5 and c-Myc affect bladder cancer progress is unknown. Herein, we explore the above points and discuss deeply its' potential mechanism. METHOD: 5637 and T24 cells were study subjects in vitro. Western blot was used to examined the protein expression. CCK8 and transwell assay were used to analyze proliferation and invasion ability. Additionally, xenograft tumor model was established. Mice imaging experiment, Immunochemistry assay and western blot were carried out. RESULT: Western blot result showed successful transfection of PRMT5-siRNA and c-Myc-siRNA. PRMT5-siRNA could inhibit c-Myc expression, and decrease the proliferation and invasion of bladder cells. And c-Myc overexpression could reverse inhibitory action caused by PRMT5 silence. And in vitro studies found low-expression of c-Myc reduced proliferation and invasion of tumor cells and make the NF-κB pathway inactivation. In vivo studies also demonstrated that inhibiting PRMT5 could downregulate c-Myc expression and inhibit the bladder cancer progress, and the potential mechanism was likely to be related to NF-κB signaling pathway. CONCLUSION: In a word, low-expression of PRMT5 suppressed c-Myc, and thus inhibited proliferation and invasion ability of 5637 and T24 cells through NF-κB pathway.

Analysis of the binding modes of the first- and second-generation antiandrogens with respect to F876L mutation.

Androgen receptor (AR) is an important target for the treatment of prostate cancer, and mutations in the AR have an important impact on the resistance of existing drugs. In this work, we performed molecular dynamics simulations of the existing marketed antiandrogens flutamide, nilutamide, bicalutamide, enzalutamide, apalutamide, darolutamide, and its main metabolite ORM15341 in complex with the wild-type and F876L mutant AR. We calculated the residue-specific binding free energy contribution of the wild-type and mutant ARs with the AS-IE method and analyzed the hotspot residues and the binding free energy contributions of specific residues before and after the mutation. In addition, we analyzed the total binding obtained by adding residue binding energy contributions and compared the results with experimental values. The obtained residue-specific binding information should be very helpful in understanding the mechanism of drug resistance with respect to specific mutations and in the design of new generation drugs against possible new mutations.

Long non-coding DANCR targets miR-185-5p to upregulate LIM and SH3 protein 1 promoting prostate cancer via the FAK/PI3K/AKT/GSK3ß/snail pathway.

BACKGROUND: Long non-coding RNA differentiation antagonizing non-protein coding RNA (DANCR) acts as an oncogene in different cancers, although its roles in prostate cancer are not fully reported. We aimed to explore its mechanism in facilitating the malignancy of prostate cancer. METHODS: The expression of DANCR, microRNA (miR)-185-5p and LIM and SH3 protein 1 (LASP1) in 40 pairs of prostate cancer tissues and normal tissues, five prostate cancer cell lines and one epithelial cell line was assessed by a quantitative real-time polymerase chain reaction, western blotting and immunohistochemistry, respectively. In transfected PC3 and C4-2 cells, cell proliferation, migration, invasion, cell cycle distribution and epithelial-mesenchymal transition (EMT) protein expression were tested via cell counting kit-8, wound healing, transwell, flow cytometry and western blot assays, respectively. The interactions between DANCR, miR-185-5p and LASP1 were verified by a dual-luciferase reporter assay. Rescue experiments were conducted to determine the roles of DANCR on the malignant properties of PC3 and C4-2 cells. The involvement of the signaling pathway was examined using a p-FAK inhibitor. RESULTS: DANCR and LASP1 expression was enhanced, whereas miR-185-5p expression was diminished in prostate cancer tissues and cell lines. Knockdown of DANCR suppressed cell proliferation, migration, invasion, G1-S transition and expression of EMT proteins of the transfected PC3 and C4-2 cells. DANCR sponged miR-185-5p to upregulate LASP1 expression. DANCR-miR-185-5p-LASP1 axis activates the FAK/PI3K/AKT/GSK3ß/Snail pathway to promote the malignant properties of PC3 and C4-2 cells. CONCLUSIONS: These findings suggest that DANCR exerts oncogenic roles in prostate cancer via the miR-185-5p/LASP1 axis activating the FAK/PI3K/AKT/GSK3ß/Snail pathway. It can be a potential biomarker in the diagnosis and monitoring of prostate cancer.

Computational Analysis of Residue-Specific Binding Free Energies of Androgen Receptor to Ligands.

Androgen receptor (AR) is an important therapeutic target for the treatment of diseases such as prostate cancer, hypogonadism, muscle wasting, etc. In this study, the complex structures of the AR ligand-binding domain (LBD) with fifteen ligands were analyzed by molecular dynamics simulations combined with the alanine-scanning-interaction-entropy method (ASIE). The quantitative free energy contributions of the pocket residues were obtained and hotspot residues are quantitatively identified. Our calculation shows that that these hotspot residues are predominantly hydrophobic and their interactions with binding ligands are mainly van der Waals interactions. The total binding free energies obtained by summing over binding contributions by individual residues are in good correlation with the experimental binding data. The current quantitative analysis of binding mechanism of AR to ligands provides important insight on the design of future inhibitors.

Real-time transrectal ultrasound-guided seminal vesiculoscopy for the treatment of patients with persistent hematospermia: A single-center, prospective, observational study.

This study aimed to describe endoscopic anatomy of the seminal tract and summarize our experience of transutricular seminal vesiculoscopy (TSV) guided by real-time transrectal ultrasonography (TRUS) in managing persistent hematospermia. A total of 281 consecutive patients with persistent hematospermia who underwent TSV with or without real-time TRUS were enrolled in this single-center, prospective, observational study. The median follow-up period was 36.5 (range: 8.0-97.5) months. TSV was successfully performed in 272 (96.8%) patients. The approach of a 4.5/6 F rigid vesiculoscope entering the seminal tract was categorized into four types on the basis of endoscopic presentation of the ejaculatory duct orifice and verumontanum. Seven (2.6%), 74 (27.2%), 64 (23.5%), and 127 (46.7%) patients had Types I (through the ejaculatory duct in the urethra), II (through the ejaculatory duct in the prostatic utricle), III (transutricular fenestration through a thin membrane), and IV (real-time transrectal ultrasound-guided transutricular fenestration) approach, respectively. In patients who successfully underwent surgery, bleeding occurred in the seminal vesicle in 249 (91.5%) patients. Seminal vesiculitis, calculus in the prostatic utricle, calculus in the ejaculatory duct, calculus in the seminal vesicle, prostatic utricle cysts, and seminal vesicle cysts were observed in 213 (78.3%), 96 (35.3%), 22 (8.1%), 81 (29.8%), 25 (9.2%), and 11 (4.0%) patients, respectively. Hematospermia was alleviated or disappeared in 244 (89.7%) patients 12 months after surgery. Fifteen patients had recurrent hematospermia, and the median time to recurrence was 7.5 (range: 2.0-18.5) months. TSV guided by TRUS may contribute to successful postoperative outcomes in managing persistent hematospermia.

Induced Intermediate Mesoderm Combined with Decellularized Kidney Scaffolds for Functional Engineering Kidney.

Background: Chronic kidney disease is a severe threat to human health with no ideal treatment strategy. Mature mammalian kidneys have a fixed number of nephrons, and regeneration is difficult once they are damaged. For this reason, developing an efficient approach to achieve kidney regeneration is necessary. The technology of the combination of decellularized kidney scaffolds with stem cells has emerged as a new strategy; however, in previous studies, the differentiation of stem cells in decellularized scaffolds was insufficient for functional kidney regeneration, and many problems remain. Methods: We used 0.5% sodium dodecyl sulfate (SDS) to produce rat kidney decellularized scaffolds, and induce adipose-derived stem cells (ADSCs) into intermediate mesoderm by adding Wnt agonist CHIR99021 and FGF9 in vitro. The characteristics of decellularized scaffolds and intermediate mesoderm induced from adipose-derived stem cells were identified. The scaffolds were recellularized with ADSCs and intermediate mesoderm cells through the renal artery and ureter. After cocultured for 10 days, cells adhesion and differentiation was evaluated. Results: Intermediate mesoderm cells were successfully induced from ADSCs and identified by immunofluorescence and Western blotting assays (OSR1 + , PAX2 +). Immunofluorescence showed that intermediate mesoderm cells differentiated into tubular-like (E-CAD + , GATA3 +) and podocyte-like (WT1 +) cells with higher differentiation efficiency than ADSCs in the decellularized scaffolds. Comparatively, this phenomenon was not observed in induced intermediate mesoderm cells cultured in vitro. Conclusion: In this study, we demonstrated that intermediate mesoderm cells could be induced from ADSCs and that they could differentiate well after cocultured with decellularized scaffolds.

GCN5 inhibition prevents IL-6-induced prostate cancer metastases through PI3K/PTEN/Akt signaling by inactivating Egr-1.

General control non-derepressible 5 (GCN5) is ectopically expressed in different types of human cancer and association with the carcinogenesis, development, and poor prognosis of cancers. The present study was aimed to investigate the potential role and related mechanisms of GCN5 in IL-6-treated prostate cancer (PCa) cell. The results showed that an elevated GCN5 expression was stimulated by IL-6. Knockdown of GCN5 significantly inhibited IL-6-driven proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT). Moreover, early growth response-1 (Egr-1) expression was elevated by IL-6 treatment and GCN5 siRNA down-regulated the expression of Egr-1. Furthermore, overexpression of Egr-1 attenuated the effects of GCN5 silence on cell proliferation, migration, invasion, and EMT in PCa. Besides, knockdown of GCN5 resulted in the down-regulation of p-Akt and up-regulation of PTEN, which was partly impeded by Egr-1 overexpression. The effects of GCN5 overexpression on cell proliferation and invasion were suppressed by LY294002, In conclusion, these data demonstrated the negative effect of up-regulated GCN5 in IL-6-induced metastasis and EMT in PCa cells through PI3K/PTEN/Akt signaling pathway down-regulating Egr-1 expression.

Uretero-fallopian fistula after hysteroscopy fallopian tube embolization: a case report.

BACKGROUND: Uretero-fallopian fistula (UFF) is a very rare surgery complication which usually happens after surgeries of fallopian tube or ureter. There has been no report of interventional operations of fallopian tube causing UFF. CASE PRESENTATION: A 41-year-old female received fallopian tube embolization for birth control. After that she noticed "clear vaginal discharge". She neglected that symptom for 7 years, until a sudden onset of abdominal pain brought her to the ER. Retrograde ureterogram confirmed UFF and revealed severe hydronephrosis of the left kidney. She received left nephrectomy afterwards and recovered well, with no urine leakage from her vagina. CONCLUSION: UFF could be caused by interventional operations of fallopian tube, and could lead to severe consequences. The application of fallopian tube embolization should be carefully controlled.

[Real-time transrectal ultrasonography in seminal vesiculoscopy].

OBJECTIVE: To compare three different pathways for transurethral seminal vesiculoscopy (SVS) and investigate the reliability and efficiency of transrectal ultrasonography (TRUS)-guided SVS (TRUS-SVS). METHODS: We retrospectively analyzed 90 cases of seminal vesiculoscopy conducted directly through the ejaculatory duct or prostatic utricle or under the guide of TRUS. We compared the success rate and complications among the three approaches. RESULTS: Operations were successfully performed in 87 (96.67%) of the 90 cases, 30 through the ejaculatory duct, 37 via the prostatic utricle, and 20 under the guide of TRUS, the operation time ranging from 25 to 75 minutes. Sperm was detected from the seminal vesicle fluid in (92.06%) of the azoospermia patients (58/63) during the surgery and in 77.78% of them (49/63) in semen analysis at 1 week postoperatively. Fifteen hematospermia and 12 spermatocystitis patients were cured. Postoperative follow-up found 20 cases of water-like semen and 3 cases of orchiepididymitis, but no such complications as retrograde ejaculation, incontinence, or rectourethral fistula. CONCLUSIONS: Transejaculatory duct and transprostatic utricle pathways are two common approaches to SVS, while TRUS-SVS may achieve a higher success rate and avoid injury of both the prostate and the rectum.

Suppression of LASP-1 attenuates the carcinogenesis of prostatic cancer cell lines: Key role of the NF-κB pathway.

Prostate cancer (PCa) is one of the most frequently diagnosed cancers among males worldwide and causes a considerable number of deaths each year. One of the newly explored targets for the development of therapies against PCa is LIM and SH3 protein 1 (LASP-1). In the present study, the function of LASP-1 in the oncogenesis and metastasis of PCa was investigated using a series of in vitro experiments. Moreover, the mechanism through which LASP-1 exerted its effect on the carcinogenesis of PCa was also explored. The expression levels of LASP-1 in clinical PCa specimens were determined both at the mRNA and protein levels. Afterwards, the activity of LASP-1 in human PCa cell lines PC3 and DU145 was inhibited using a short hairpin RNA (shRNA) interfering method. The effects of LASP-1 knockdown on the cell growth, apoptosis, cell cycle distribution, migration and invasion were assessed. It was demonstrated that the expression of LASP-1 was significantly higher in the clinical PCa tissues than the level in the corresponding para-carcinoma tissues. Following the knockdown of the LASP-1 gene in human PCa cell lines, the viability, migration and invasion of the cancer cells were decreased. It was also demonstrated that the change in the cell viability and motile ability were associated with an induction of cell apoptosis and G1 phase cell cycle arrest. Based on the results of the detection of the expression of NF-κB-related factors, it was indicated that LASP-1 may affect the carcinogenesis of PCa through a NF-κB inhibition-dependent manner. Although the detailed explanation of the mechanism of LASP-1 in the carcinogenesis of PCa requires further elucidation, the present study highlights the potential of LASP-1 as a promising therapeutic target to ameliorate the oncogenesis and metastasis of PCa.

[Narrow band imaging-assisted holmium laser resection reduces the residual tumor rate of primary non-muscle invasive bladder cancer: a comparison with the standard approach].

OBJECTIVE: To assess the feasibity and efficacy of narrow band imaging (NBI) cystoscopy assisted holmium laser resection of primary non-muscle invasive bladder cancer (HoLRBt). METHODS: During the period of May 2013 to December 2014, 150 cases of primary non-muscle invasive bladder cancer (NMIBC) admitted in our hospital were randomly divided into NBI-HoLRBt and WLI-TURBt group. In NBI-HoLRBt group, all suspicious lesion identified by either WLI or NBI were resected during the surgery with WLI and in NBI mode for lesion only visible with NBI. At the end of the procedure, NBI cystoscopic examination was performed again to identify whether there was residual lesions at the margins of the resection areas. In WLI-TURBt group, only WLI and TURBt were applied. All patients from the two groups underwent routine intravesical instillation after surgery. A total of 124 patients were diagnosed NMIBC by pathological findings (NBI-HoLRBt group: n=60, WLI-TURBt group: n=64), they were followed-up at 3 months, at which both WLI and NBI cystoscopy were performed to examine the residual tumor, and cytology was checked for all patients. The residual tumor rates at the first follow-up (RR-fFU) were recorded and compared. RESULTS: Baseline characteristics of the patient and the tumor were comparable between the two groups. The overall detection rate of NMIBC and carcinoma in situ (CIS) were significantly higher with NBI than WLI (94.5% (137/145) vs 75.8% (110/145), 16/17 vs 10/17, both P<0.05). The RR-fFU for NBI-HoLRBt and WLI-TURBt was 3.3% (2/60) and 17.2% (11/64), respectively (P<0.05). CONCLUSION: NBI-HoLRBt was feasible, and more effective for identification of NMIBC as well as for the reduction of residual tumor rate compared with WLI-TURBt.

Outcomes of Microscopic Subinguinal Varicocelectomy With and Without the Assistance of Doppler Ultrasound: A Randomized Clinical Trial.

OBJECTIVE: To compare the surgical outcomes and complications between microscopic subinguinal varicocelectomy (MV) and intraoperative vascular Doppler ultrasound-assisted microscopic subinguinal varicocelectomy (IVDU-MV) for infertile patients with varicoceles. MATERIALS AND METHODS: One hundred seventy-two infertile patients with varicoceles were randomly divided into IVDU-MV group (n = 85) and MV group (n = 87). We assessed patients' operative and postoperative parameters, semen parameters, and the pregnancy rate. The mean follow-up period was 21 months (range, 13-34 months). RESULTS: The operative time was significantly shorter in the IVDU-MV group than MV group (41.9 ± 13.6 vs 52.7 ± 14.1 minutes, P <.05). The number of intraoperative arteries spared was significantly greater in the IVDU-MV group than the MV group (1.9 ± 0.8 vs 1.3 ± 0.7, P <.05). In addition, the average number of spermatic veins ligated was significantly greater in the IVDU-MV group (7.8 ± 2.1 vs 7.0 ± 1.9, P <.05). Lymphatic spared showed no significant difference (P >.05). The postoperative hospital stay showed no significant difference. Sperm concentration, sperm motility, and the percentage of grade a+b sperm were significantly increased in both groups at 3, 6, and 12 months after surgery (P <.05), and the sperm motility was higher in IVDU-MV than MV group (43.98 ± 7.64 vs 36.98 ± 5.10, P <.05) in 12 months after surgery. Sperm morphology was comparable between the 2 groups. The pregnancy rate showed no significant difference (36.8% of the MV vs 34.1% of the IVDU-MV, P >.05). CONCLUSION: Our study demonstrated that both MV and IVDU-MV are effective methods for the improvement of semen parameters in infertile men with varicocele, with a natural conception rate of 35% over a mean follow-up of 21 months. Compared with MV, IVDU-MV is superior in shortening operative time, increasing the number of spermatic arteries spared, spermatic veins ligated, and sperm motility after surgery. IVDU should be routinely used as an effective tool to improve outcomes and safety of varicocelectomy.

[Narrow band imaging-assisted holmium laser resection reduced the recurrence rate of non-muscle invasive bladder cancer: a prospective, randomized controlled study].

OBJECTIVE: To compare the differential effects of narrow band imaging (NBI)-assisted holmium laser with transurethral resection on the 1-year recurrence rate of non-muscle invasive bladder cancer (NMIBC), and to evaluate the clinical values of NBI-assisted holmium laser resection for NMIBC (NBI-HoLRBt). METHODS: During the period of February 2013 to February 2014, 178 cases of NMIBC were randomly divided into NBI-HoLRBt group and white light imaging (WLI) assisted transurethral resection of bladder tumor (WLI-TURBt) group. In NBI-HoLRBt, all suspicious lesion identified by either WLI or NBI were resected with WLI and in NBI mode for lesion only visible with NBI. At the end of the procedure, a NBI cystoscopic examination was performed to assess the margins of the resection areas and to identify eventual residual lesions. In WLI-TURBt group, only WLI and TURBt were applied. All patients underwent routine follow-up with WLI and NBI cystoscopy supplemented with cytology every 3 month. The recurrence risk of patients with NMIBC subjected to either NBI-HoLRBt or WLI-TURBt was compared at 3 and 12 month. RESULTS: The 3-month and 1-year recurrence rate was 18.48% (17/92) and 38.04% (35/92) respectively in the WLI-TURBt group, it was 5.81% (5/86) and 18.60% (16/86) in the NBI-HoLRBt group (both P<0.05). In addition, the in situ recurrence rate was less in the NBI-HoLRBt than WLI-TURBt group (2.33% vs 14.13%, P<0.05). CONCLUSION: NBI-assisted holmium laser resection of bladder tumor can reduce the 3-month and 1-year recurrence risk of NMIBC and should be considered a valuable clinical therapeutic method for NMIBC.