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Bladder cancer is one of the most severe genitourinary cancers, causing high morbidity worldwide. However, the underlying molecular mechanism is not clear, and it is urgent to find target genes for treatment. G-protein-coupled receptors are currently a target of high interest for drug design. Thus, we aimed to identify a target gene-related to G-protein-coupled receptors for therapy. We used The Cancer Genome Atlas (TCGA) and DepMap databases to obtain the expression and clinical data of RGS19. The results showed that RGS19 was overexpressed in a wide range of tumor, especially bladder cancer. We also explored its effect on various types of cancer. High expression of RGS19 was also shown to be significantly associated with poor prognosis. Cell models were constructed for cell cycle detection. shRGS19 can halt the cell cycle at a polyploid point. RGS19 is a G-protein-coupled receptor signaling pathway-related gene with a significant effect on survival. We chose RGS19 as a therapeutic target gene in bladder cancer. The drug GSK1070916 was found to inhibit the effect of RGS19 via cell rescue experiments in vitro.

Intravascular migration of a double J stent into the inferior vena cava is an uncommon complication. Active prevention, timely diagnosis, and early intervention are crucial for this complication. Intravascular interventional therapy is relatively easy, less traumatic, and has a high success rate. It can be used to select patients for intravascular ectopic DJS treatment., Intravascular interventional therapy can be used to select patients to retrieve the ectopic intravascular DJS because it is relatively easy, less invasive, and has a high success rate.

Endothelial cells (ECs) play a key role in revascularization within regenerating tissue. Stem cells are often used as an alternative cell source when ECs are not available. Several cell types have been used to give rise to ECs, such as umbilical cord vessels, or differentiated from somatic stem cells, embryonic, or induced pluripotent stem cells. However, the latter carry the potential risk of chronic immune rejection and oncogenesis. Autologous endothelial precursors are an ideal resource, but currently require an invasive procedure to obtain them from the patient's own blood vessels or bone marrow. Thus, the goal of this study was to determine whether urine‐derived stem cells (USCs) could differentiate into functional ECs in vitro. Urine‐derived cells were then differentiated into cells of the endothelial lineage using endothelial differentiation medium for 14 days. Changes in morphology and ultrastructure, and functional endothelial marker expression were assessed in the induced USCs in vitro. Grafts of the differentiated USCs were then subcutaneously injected into nude mice. Induced USCs expressed significantly higher levels of specific markers of ECs (CD31, vWF, eNOS) in vitro and in vivo, compared to nondifferentiated USCs. In addition, the differentiated USC formed intricate tubular networks and presented similar tight junctions, and migration and invasion ability, as well as ability to produce nitric oxide (NO) compared to controls. Using USCs as autologous EC sources for vessel, tissue engineering strategies can yield a sufficient number of cells via a noninvasive, simple, and low‐cost method suitable for rapid clinical translation. Stem Cells Translational Medicine 2018 Stem Cells Translational Medicine 2018;7:686–698

Arginine methylation is mainly catalyzed by protein arginine methyltransferases (PRMTs) and is one of the most common posttranslational modifications closely related to the development of cancer. PRMT7 is overexpressed in various tumors and promotes the malignant progression of tumors, but the expression and role of PRMT7 in renal cell carcinoma (RCC) remains unclear. Here, we report for the first time that the expression of PRMT7 is increased in clear cell renal cell carcinoma (ccRCC) tissues and that it may act as an independent predictor for the poor prognosis of ccRCC patients. We found that PRMT7 promotes RCC cell proliferation both in vitro and in vivo. Moreover, the methyltransferase inhibitor adenosine dialdehyde (Adox) blocks the action of PRMT7 in ccRCC cells. Furthermore, PRMT7 regulates the expression of C-MYC, which plays an important role in promoting ccRCC cell proliferation, and it accelerates the tumor development of RCC in a C-MYC-dependent manner. Mechanistically, PRMT7 upregulates the expression of C-MYC via methylating β-catenin and inhibiting the ubiquitin-mediated degradation of β-catenin. In conclusion, our study demonstrates that overexpressed PRMT7 in ccRCC cells acts as an oncogene to promote the growth of renal cell carcinoma through regulating the β-catenin/C-MYC axis, thereby providing new strategies and targets for the treatment of ccRCC patients.

Background Autologous urothelial cells are often obtained via bladder biopsy to generate the bio-engineered urethra or bladder, while urine-derived stem cells (USC) can be obtained by a non-invasive approach. The objective of this study is to develop an optimal strategy for urothelium with permeability barrier properties using human USC which could be used for tissue repair in the urinary tract system. Methods USC were harvested from six healthy adult individuals. To optimize urothelial differentiation, five different differentiation methods were studied. The induced cells were assessed for gene and protein expression markers of urothelial cells via RT-PCR, Western blotting, and immunofluorescent staining. Barrier function and ultrastructure of the tight junction were assessed with permeability assays and transmission electron microscopy (TEM). Induced cells were both cultured on trans-well membranes and small intestinal submucosa, then investigated under histology analysis. Results Differentiated USC expressed significantly higher levels of urothelial-specific transcripts and proteins (Uroplakin III and Ia), epithelial cell markers (CK20 and AE1/AE3), and tight junction markers (ZO-1, ZO-2, E-cadherin, and Cingulin) in a time-dependent manner, compared to non-induced USC. In vitro assays using fluorescent dye demonstrated a significant reduction in permeability of differentiated USC. In addition, transmission electron microscopy confirmed appropriate ultrastructure of urothelium differentiated from USC, including tight junction formation between neighboring cells, which was similar to positive controls. Furthermore, multilayered urothelial tissues formed 2 weeks after USC were differentiated on intestine submucosal matrix. Conclusion The present study illustrates an optimal strategy for the generation of differentiated urothelium from stem cells isolated from the urine. The induced urothelium is phenotypically and functionally like native urothelium and has proposed uses in in vivo urological tissue repair or in vitro urethra or bladder modeling. Electronic supplementary material The online version of this article (10.1186/s13287-018-1035-6) contains supplementary material, which is available to authorized users.

// Gaosi Xu 1, * , Xiongbing Lu 2, * , Tianlun Huang 1 , Jie Fan 1 1 Department of Nephrology, The Second Affiliated Hospital of Nanchang University, Nanchang, China 2 Department of Urology, The Second Affiliated Hospital of Nanchang University, Nanchang, China * These authors have contributed equally to this work Correspondence to: Gaosi Xu, email: gaosixu325@163.com Keywords: renal cancer, ARHGAP24, proliferation, cell cycle, apoptosis Received: March 14, 2016 Accepted: June 13, 2016 Published: July 2, 2016 ABSTRACT Renal cell carcinoma (RCC) is the major cause of kidney malignancy-related deaths. Rho GTPases are key regulators in cancer cell metastasis. ARHGAP24, a Rac-specific member of the Rho GTPase-activating protein family, acts as a functional target of cancer cell migration and invasion. In the present study, we identified ARHGAP24 expression is downregulated in renal cancer tissues and is highly correlated with long-term survival in RCC patients. Therefore, we investigated the biological functions of ARHGAP24 in renal cancer cells. Ectopic expression of ARHGAP24 resulted in inhibited cell proliferation and arrested cell cycle in two renal cancer cell lines (786-0 and Caki-2); the results were confirmed by ARHGAP24 knocking down. In addition, ARHGAP24 significantly reduced the cell invasion ability and induced apoptosis in renal cancer cells. In addition, overexpressing ARHGAP24 impaired tumor formation in vivo . In summary, our results illustrated that ARHGAP24 plays a unique role in RCC progression as a tumor repressor.

Urinary bladder cancer (UBC) is one of the most common malignancies worldwide. UBC patients at muscle invasive stage have poor clinical outcome, due to high propensity for metastasis. Non-tumor activated fibroblasts, named α-SMA+Fs, is similar to carcinoma-associated fibroblasts (CAFs) which could express α-SMA. However, whether α-SMA+Fs patients could induce UBC cell invasion is unclear. Herein, we found that characterization of primary α-SMA+Fs separated from PBOO (partial bladder outlet obstruction) rats was fell in between normal fibroblasts (α-SMA-Fs) and CAFs. Additionally, the conditional medium from α-SMA+Fs enhanced the NBT-II cell invasion through inducing EMT, and the oncogenic function of mixed supernatant of α-SMA+Fs/CAFs was stronger than that of CAFs. Inhibition of TGF-β1 by TGF-β1 neutralizing antibody decreased the EMT-associated gene expression and NBT-II cell invasion, suggesting that α-SMA+Fs can induce tumor EMT through TGF-β1. Xenograft experiments showed that the tumorigenic effect of α-SMA+Fs in mice was also between CAFs and α-SMA-Fs, and α-SMA+Fs/CAFs also had a strong tumorigenic effect. We preformed rats with PBOO and found that the incidence of invasive bladder cancer in PBOO+BBN group was higher than in BBN group, suggesting the PBOO treatment contributed to tumorigenesis. Thus, α-SMA+Fs promoted tumorigenesis by secreting TGF-β1 to induce EMT.

Increasing reports indicate that circular RNAs (circRNAs) are very important regulators in human diseases, including cancers. In bladder cancer (BC), several circRNAs have been reported to be involved in tumor progressions, such as circ-ITCH and circTCF25. However, the functions of most circRNAs in BC still remains largely unknown. In this study, we identified a novel circRNA termed as circ-VANGL1 by bioinformatics analysis. We found that circ-VANGL1 was highly expressed in BC tissues compared with adjacent normal tissues. Furthermore, we showed that circ-VANGL1 could serve as a prognostic marker for patients with BC. Through functional experiments, we found that circ-VANGL1 knockdown significantly suppressed BC cell proliferation, cell cycle, migration, and invasion in vitro. Besides, circ-VANGL1 silence inhibited BC cell propagation in vivo. Mechanistically, we identified circ-VANGL1 as a sponge of miR-605-3p which targeted VANGL1 in BC cells. Through repressing miR-605-3p availability, circ-VANGL1 contributes to VANGL1 expression, consequently leading to BC cell proliferation, migration, and invasion. Taken together, our study demonstrated circ-VANGL1/miR-605-3p/VANGL1 as a novel essential signaling pathway involved in BC progression.

Table S4. Percentage of USC expressing tight junction markers 2Â weeks after urothelial induction assessed by immunofluorescence. (DOCX 16Â kb)

Human urine-derived stem cells (USCs), a subpopulation of cells isolated from urine, possess biological characteristics of stem cells, i.e., clonogenicity, cell growth patterns and expansion capacity, cell surface marker expression profiles, capacity for multipotent differentiation, paracrine effects, and immunomodulatory properties. USCs originate from the parietal epithelial cells in kidney glomeruli. These cells not only can differentiate into mesenchymal cell lineages, but also renal cells, including podocytes and endothelial cells. USCs possess immunomodulatory properties via inhibition of T-cell and B-cell proliferation. In vivo studies demonstrated that USCs significantly enhanced renal function via antiinflammatory and antifibrotic mechanisms after implantation of USCs in rodent models of renal function insufficiency. Importantly, USCs can be obtained via a simple, noninvasive and low-cost approach. Thus, USCs provide a very promising cell source for treatment of renal diseases.

Ectopic insertion of the ureter in the genitourinary tract is a rare congenital disorder, usually associated with ureteral duplication. Identification of the insertion open is critical for ureteric re-implantation. However, the challenge in the diagnosis of ectopic insertion of the ureter usually is to identify its insertion, particularly when the affected ureter is not dilated. Multi-detector computed tomography (MDCT) urography with nonionic iodinated contrast media delineates the ureteric course in the normal functioning kidney in the excretory phase [1]. This report presented a young male patient with ectopic insertion of a duplicated ureter diagnosed by MDCT urography. Three-dimensional (3D) analysis technology, such as volume rendering (with a color display improving the visualization of complex anatomy and 3D relationships) and maximum intensity projection (similar in principle to projection angiography), is useful for the illustration of urinary tract anatomy [1]. Rotated volume rendering reconstruction images and continual thinner maximum intensity projection reformatted images can be viewed as videos, which provides detail delineation of the ectopic ureteral insertion and its associated ureteral duplication.In this study, we reported MDCT urography and 3D analysis technology as an appropriate diagnostic method for the ectopic ureteral insertion and its associated complications.

Noncontrast computed tomography (CT) has great advantage with higher sensitivity and more clear modalities in detecting urinary tract radiolucent calculi in patients with acute renal insufficiency (ARI) compared to other image diagnosis approaches. We report two cases (female, 28 years old; male, 39 years old) with persistent flank pain and acute anuria after the administration of ceftriaxone (4.0 g daily) for 2 days intravenously. No abnormality was found in the kidney-ureter- bladder (KUB) areas with plain abdomen X-rays. A diagnosis of bilateral hydronephrosis was made by ultrasound examination in both cases. Serum creatinine levels reached up to 257 and 810 μ mol/L (normal serum creatinine level is 40-130 μ mol/L), respectively. Vague density spots were noticed in the pelvis with noncontrast multidetector-row CT (MDCT) scanning. However, distinguishable clusters of high-density shadows were seen in pelvic areas with maximum intensity projections (MIP, CT values in 30-128 HU). Ceftriaxone crystal calculi were found on both sides of distal ureters under endoscopy. Renal function recovered in both patients after double-J ureteral stents were installed. Out results demonstrated that noncontrast MDCT scanning and MIP reconstruction as an effective diagnostic tool could provide clear images in detection of radiolucent calculi in urinary tract when conventional X-rays image are not suitable in the patients with obstructive anuria and ARI of unknown origin.

Objective: This study was designed to investigate the effect of chromium (VI) on DNA-protein crosslinks (DPC) of SPC-A1 cells. Methods: We exposed SPC-A1 cells were cultured in 1640 medium and treated with the SPC-A1 cells in vitro to different concentrations of Hexavalent chromium Cr(VI) for 2h, the KC1-SDS precipitation assay were used to measure the DNA-protein cross-linking effect. Results: All the different concentrations of Cr(VI) could cause the increase of DPC coefficient in SPC-A1 cells. But this effect was not significant (P>0.05) at low concentrations; while in high concentration Cr(VI) induced SPC-A1 cells could produce DNA-protein cross-linking effect significantly (P

Tissue-specific stem cells can more efficiently repair the tissue from which they originate, compared to other types of adult stem cells. Human urine-derived stem cells (USCs), a subpopulation of cells constantly exposed to urine, possess biological characteristics of stem cells, and thus are an optimal cell source for tissue repair in the genitourinary system. USCs display clonogenicity, high proliferation capacity, stem cell marker expression profiles, and multipotent differentiation capacity. These cells can more efficiently differentiate into the cell types of the genitourinary tract, particular in urothelial cells, renal podocytes, and endothelial cells, compared to other adult stem cells. In addition, USCs secrete numerous growth factors and cytokines to support cell growth, differentiation, resident stem cell recruitment, and immunomodulatory properties. In vivo studies in rodent models of diabetic erectile dysfunction, urinary tract incontinence, and renal dysfunction demonstrated that USCs significantly enhanced organ function via angiogenesis and transdifferentiation, and showed anti-inflammatory and anti-fibrotic properties after implantation of USCs. USCs can be obtained via a simple, non-invasive, and low-cost approach. Thus, USCs offer an extremely promising cell source for cell-based therapy in androurology.

Objective: To observe the lethal effect of T cells activated by dendritic cells (DCs) with MUC1 gene transfection (MUC1-DCs) on BIU-87 cells in vitro, and to evaluate the possibility of dendritic cells transfected by MUC1 gene as a vaccine for bladder cancer immunotherapy. Methods: MUC1 was successfully transfected into cultured human blood-derived dendritic cell with lipofectin. The transfection efficiency and immunocompetence were detected by flow cytometry and MTT colourmetry. T lymphocytes activated by MUC1-DCs were used to kill BIU-87 cell lines and normal bladder epithelium in vitro and the killing rate was evaluated by MTT colourmetry. Results: There was significant lethal effect on the BIU-87 cells and normal bladder epithelium with T cells activated by MUC1-transfected DCs, but the lethal effect on the BIU-87 cells significantly exceeded that on normal bladder epithelium (P

As a tubularized organ in the distal portion of urinary tract system, urethras may have strictures often due to injury, infections, and congenital defects. Two types of tissue engineering technologies have been used to build urethral tissue, namely cell-seeded and non-seeded. Cell-seeded technology uses scaffolds that are seeded in vitro with primary cultured cells obtained from a bladder or oral mucosa biopsy. This composite graft is then implanted back in the host to continue the regenerative process. The non-seeded technology uses cell-free biodegradable scaffolds to allow the natural process of regeneration to occur in vivo and was successfully applied experimentally in partial (patch tissue replacement) urethral reconstruction models. Increasing evidence has demonstrated that use of autologous cells seeded on biodegradable scaffolds achieved better outcomes in urethral tissue regeneration, particularly in long segment of urethral replacement. Stem cells obtained from patients’ own tissues provide an alternative for cell therapy and tissue engineering in urology. Our recent study displayed that a subpopulation of cells isolated from naturally voided urine possesses the features of progenitor/stem cells and has potential as an alternative cell source for tissue engineering and cell therapy in urology. These cells adhere to plastic, are clonogenic, express mesenchymal stem cells/pericyte cell surface markers, and display high telomerase activity. In addition, they possess high expansion capability and multipotential differentiation capacity. Importantly, these cells can give rise to urothelium, smooth muscle to form urethral tissue in vivo in a highly efficient manner when seeded on porous natural collagen-based scaffolds. In this chapter, we discuss the recent progress made in urethral tissue engineering with urine-derived stem cells and acellular matrix for potential use in clinical applications.

Table S2. Antibodies used in this study. (DOCX 14Â kb)

Table S2. Antibodies used in this study. (DOCX 14Â kb)

Table S3. Percentage of USC expressing urothelial cell markers 2Â weeks after urothelial induction assessed by immunofluorescence. (DOCX 14Â kb)