Upregulation of Cytotoxic T-Lymphocyte-Associated Protein 4 and Forkhead Box P3 Transcripts in Peripheral Blood of Patients with Bladder Cancer.

Background: Regulatory T cells (Tregs) play a key role in the progression of tumors. These cells express forkhead box P3 (FOXP3) and cytotoxic T-lymphocyte-associated protein 4 (CTLA4), which are the potential targets for cancer immunotherapy. The present study aimed to evaluate FOXP3 and CTLA4 transcripts in patients with bladder cancer (BC) compared with healthy individuals. Methods: Transcripts of CTLA4 and FOXP3 genes in the peripheral blood mononuclear cells (PBMCs) of 50 patients with histologically confirmed BC and 50 healthy individuals were assessed at the Institute for Cancer Research, Shiraz University of Medical Sciences (Shiraz, Iran) during 2014-2016. RNA was extracted from PBMCs, then cDNA was synthesized and subjected to quantitative real-time PCR (qRT-PCR) using appropriate primers. Statistical analysis was performed using SPSS software (version 21.0). Results: Significantly higher amounts of CTLA4 and FOXP3 gene transcripts were found in the peripheral blood of BC patients compared with healthy individuals. The expression of both genes was significantly higher in patients with non-invasive and grade I/II BC. The median of CTLA4 and FOXP3 transcript expressions was 3.74 and 5.39, respectively, in non-invasive BC patients, which was significant compared with the control group (P=0.0016 and P=0.009, respectively). The median of target gene mRNA expression in grade I/II BC patients was 2.9 for CTLA4 and 6.61 for FOXP3, which was significant compared with the controls (P=0.013 and P=0.0037, respectively). Conclusion: This study highlights the functional activity of Tregs in early stages of bladder cancer and showed the importance of CTLA4 and FOXP3, when it comes to screening BC.

Circulating Levels of M30 and M65 Molecules in Transitional Cell Carcinoma of the Bladder and Their Relation to Tumor Progression.

BACKGROUND: Various markers are suggested for diagnosis and monitoring of transitional cell carcinoma of the bladder (TCC), including cytokeratins (CKs). OBJECTIVES: In the present study, the circulating CK18 (M65) and its caspase-cleaved form, ccCK18 (M30), have been investigated in a group of patients with TCC. PATIENTS AND METHODS: Serum samples were obtained from 60 patients before surgical resection, among which the samples of 26 patients after resection were also included. We measured the levels of soluble M30 and M65 molecules by enzyme-linked immunosorbent assay. The relation between these markers and patients' clinical characteristics was evaluated. RESULTS: M30 and M65 in total patient sera were 148 ± 16 U/L and 318 ± 34 U/L, respectively. A correlation existed between pre-operative M30 and M65 levels (P < 0.0001, Spearman r = 0.51). M65, but not M30, showed a significant relation to tumor stage and grade. The M65 quantity in patients with T3/T4 tumor stages (350 ± 42 U/L) was higher than that of patients with T1/T2 stages (293 ± 45U/L; P < 0.038). Patients with tumor grades III/IV also showed higher levels of M65 compared to patients with tumor grades I/II (P < 0.04). The M30:M65 ratio in all patients was 0.54 ± 0.04. There was a lower M30:M65 ratio in patients with T3/T4 stage tumors and those with tumor grades III/IV (P < 0.02). The M30 (133 ± 19 U/L) and M65 levels (240 ± 21 U/L) after surgery did not significantly differ compared to their pre-operative values. However, a correlation between the pre- and post-operative M30:M65 ratio in patients ≥ 70 years was seen (P = 0.009). CONCLUSIONS: These data suggested a relationship of both M65 and the M30:M65 ratio to tumor progression which might imply their importance in TCC monitoring.

Increased interleukin-17 transcripts in peripheral blood mononuclear cells, a link between T-helper 17 and proinflammatory responses in bladder cancer.

BACKGROUND: Interleukin (IL)-17-producing CD4+ T helper (Th17) cells thatare known by producing IL-17 have recently been defined as a unique subset of proinflammatory helper cells. IL-17 is an inflammatory cytokine with robust effect on many cells and it can play important roles in pathogenesis of diverse groups of immune-mediated diseases. OBJECTIVES: The aim of this case-control study was to determine the gene expression of IL-6, IL-17, and transforming growth factor beta (TGF-ß) in Iranian patients with bladder cancer. PATIENTS AND METHODS: Blood samples were collected from 37 patients with bladder cancer and 37 healthy individuals with no history of malignancies or autoimmune disorders, based of simple sampling. The expression of IL-6, IL-17, and TGF-ß were measured by quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS: The mean of IL-17 transcripts was significantly higher in patients with bladder cancer compared with healthy individuals (0.33 ± 0.06 vs. 0.42 ± 0.14, ) (P = 0.04), but their TGF-ß was lower (12.53 ± 8.41 vs. 54.94 ± 17.95, ) (P = 0.04). However, the IL-6 transcripts level was similar in both groups (5.34 ± 2.40 vs. 8.07 ± 3.28, ) (P > 0.05) and there was not any significant difference between the noted cytokines expressions among patients with different stages and grades. CONCLUSIONS: As most of the cases studied in this investigation were in stages I and II, IL-17 as a prominent proinflammatory cytokine may play an important role in recruiting and infiltrating of antitumor immune responses in early stages of bladder cancer. Furthermore, it can be used as predictor for the clinical stage and prognosis of cancers such as bladder carcinoma.

Soluble CD138/Syndecan-1 Increases in the Sera of Patients with Moderately Differentiated Bladder Cancer.

BACKGROUND: CD138/Syndecan-1 (Sdc-1) is expressed on the tumor and stromal cells of invasive bladder carcinoma. CD138/Sdc-1 shedding from the cell surface is associated with the invasive phenotype in lung and breast cancers. PATIENTS AND METHODS: Soluble CD138/Sdc-1 was measured in the sera of 86 bladder cancer patients and 57 healthy individuals by a commercial ELISA assay. RESULTS: Soluble Sdc-1 was increased in the sera of patients with bladder cancer (138.42 ± 81.85 vs. 86.48 ± 82.58 ng/ml, p = 0.0003). Patients aged over 70 years had higher levels of CD138/Sdc-1 in their sera (159.7 ± 15.77 vs. 124.5 ± 9.99 ng/ml, p = 0.025), and soluble Sdc-1 levels were higher in the sera of patients with moderately differentiated tumors compared to poorly differentiated ones (170.47 ± 85.06 vs. 101.79 ± 68.24 ng/ml, p = 0.01). The soluble Sdc-1 level was higher in muscle-invasive (154.45 ± 83.60 vs. 89.9 ± 55.02 ng/ml) but not lymphatic-invasive (106.25 ± 52.10 vs. 123.43 ± 63.76 ng/ml) tumors (p = 0.027 and 0.45, respectively). A non-significant trend of soluble Sdc-1 increase in the sera of male patients compared to female patients was observed (145.38 ± 85.47 vs. 110.20 ± 59.04 ng/ml, p = 0.054). CONCLUSION: The elevated levels of soluble CD138/Sdc-1 in older bladder cancer patients and those with muscular invasion sheds some light on the mechanisms of the disease invasion.

Role of viruses in renal cell carcinoma.

To determine whether viral infections are related to renal cell carcinoma (RCC), we studied 49 patients with RCC (29 patients were males with age ranging from 30 to 81 years and a mean of 57.5 years; 20 patients were females with age ranging from 36 to 70 years with a mean of 58.4 years) and 16 non-neoplastic kidney patients as controls. Tissues specimens from study patients and controls were examined by nested polymerase chain reaction (PCR) to determine the presence of DNA of several viruses including human papilloma virus (HPV), Epstein-Barr virus (EBV), and polyoma viruses (BKV and JCV). Our results revealed that 7 of 49 (14.29%) RCC tissue specimens had HPV DNA compared with none of 16 non-cancer control subjects. Regarding the HPV types, all the positive results were high-risk HPV types (type 16 in three and 18 in four patients). The present study suggests that HPV infection, especially high-risk types, is associated with RCC. However, more studies are necessary to demonstrate the molecular oncogenic processes involved in this association.

European genome-wide association study identifies SLC14A1 as a new urinary bladder cancer susceptibility gene.

Three genome-wide association studies in Europe and the USA have reported eight urinary bladder cancer (UBC) susceptibility loci. Using extended case and control series and 1000 Genomes imputations of 5 340 737 single-nucleotide polymorphisms (SNPs), we searched for additional loci in the European GWAS. The discovery sample set consisted of 1631 cases and 3822 controls from the Netherlands and 603 cases and 37 781 controls from Iceland. For follow-up, we used 3790 cases and 7507 controls from 13 sample sets of European and Iranian ancestry. Based on the discovery analysis, we followed up signals in the urea transporter (UT) gene SLC14A. The strongest signal at this locus was represented by a SNP in intron 3, rs17674580, that reached genome-wide significance in the overall analysis of the discovery and follow-up groups: odds ratio = 1.17, P = 7.6 × 10(-11). SLC14A1 codes for UTs that define the Kidd blood group and are crucial for the maintenance of a constant urea concentration gradient in the renal medulla and, through this, the kidney's ability to concentrate urine. It is speculated that rs17674580, or other sequence variants in LD with it, indirectly modifies UBC risk by affecting urine production. If confirmed, this would support the 'urogenous contact hypothesis' that urine production and voiding frequency modify the risk of UBC.

Renal carcinoid tumor.

Primary renal carcinoid is a rare tumor, until now less than 50 cases of this tumor has been reported, so very little is known about its presentation clinicopathologic patterns and prognosis. We report a patient with primary carcinoid tumor of kidney in a middle-aged woman treated by nephrectomy and review the literature concerning this kind of neoplasm.

Application of amniotic membrane as xenograft for urethroplasty in rabbit.

OBJECTIVES: Uroepithelium repair has always been the subject of discussion in urology surgeries. Using human amniotic membrane allograft has already proved to be useful in other fields. In this study, we use amniotic membrane to repair uroepithelium injuries in rabbits. METHODS: Twenty healthy rabbits underwent surgery to induce a 10-mm incision, 10 mm from the meatus on ventral part of the urethra. Then a 5 x 10 mm patch of amniotic membrane was sutured to the incised urethra. All of the rabbits were catheterized for a week and then the patches were removed. The rabbits were studied for a month for any signs of infection and fistula formation. RESULTS: All samples revealed complete re-epithelialization of reconstructed urethra by transitional epithelium. There was one case of infection and following fistula (5%). There were two cases of urethral strictures (10%). CONCLUSION: The result from this study suggests that amniotic membrane is an inexpensive, easy, and biodegradable graft with very little antigen effect which seems to be the ideal solution for urethroplasty.

Combined use of Mathieu procedure with plate incision for hypospadias repair: a randomized clinical trial.

OBJECTIVES: To introduce a simple and effective technique for urethroplasty of distal hypospadias, using the advantages of both standard techniques of Snodgrass and Mathieu to reduce or eliminate the shortcomings of each. METHODS: Forty patients with primary distal hypospadias were randomized between two groups (20 in each). Patients in group I underwent standard Snodgrass urethroplasty, whereas those in group II were operated with our novel technique, Mathieu-Incised Plate (Mathieu-IP), comprising preparation of a parameatal-based skin flap as in the Mathieu procedure together with modifications (adopted from Snodgrass urethroplasty) such as a complete longitudinal incision on the urethral plate as well as coverage of the neourethra with an inner preputial flap. Operative data and outcomes of both procedures were analyzed prospectively after 1 year of follow-up. RESULTS: Both groups were age matched (group I: 7.05 +/- 4.85 years; group II: 7.31 +/- 2.17 years), and there were no significant differences in mean operative time (95 minutes versus 100 min in groups I and II, respectively). There were significant differences between the two groups in terms of complications. Whereas 3 cases of posturethroplasty meatal stenosis and 2 of urethrocutaneous fistula (5 complications, 25%) were detected in the Snodgrass group, neither of these complications occurred in the Mathieu-IP group. All 40 patients benefited from a slit-like meatus. CONCLUSIONS: Combining the Mathieu procedure with plate incision could be considered a promising simple technique to achieve a normally shaped meatus and to reduce the rate of meatal-related complications, the major concern with the Snodgrass procedure.

Ischemic preconditioning protects the dog kidney from ischemia-reperfusion injury.

OBJECTIVE: To evaluate the effects of unilateral ischemic insult and ischemic preconditioning (IPC) on renal histology in a canine model. METHODS: 30 dogs were randomized into 4 groups. In group A (5 male controls) and group B (5 female controls), ischemia was induced by clamping both left renal arteries for 40 min. Dogs in group C (10 male cases) or group D (10 female cases) underwent 5 min of arterial clamping and 10 min of declamping prior to the final 40-min ischemia induction. Renal biopsy was prepared 48 h later and microscopically examined. RESULTS: The control groups (A and B) developed 40% frank necrosis, 60% moderate injury, and there was no intact renal tissue in this group with no difference between sexes. The IPC groups (C and D) revealed 55% moderate injury and 45% normal pathology; however, there was no frank necrosis among them. Better IPC protection in the female group was not statistically significant. CONCLUSION: An IPC schedule of 5-min ischemia and 10-min reperfusion improves ischemia-reperfusion injury from subsequent prolonged ischemia in a canine model.

Effect of intraurethral captopril gel on the recurrence of urethral stricture after direct vision internal urethrotomy: Phase II clinical trial.

OBJECTIVE: The purpose of this study is to evaluate the effect of intraurethral captopril gel as an antifibrotic agent on patients with urethral stricture. MATERIALS AND METHODS: In the first phase of clinical trial, 13 rabbits were included and local side-effects of captopril gel were evaluated. In the second phase, 56 patients were enrolled from April 2004 to January 2006. After internal urethrotomy the patients were classified into three patient groups: (i) received placebo gel (group I); (ii) received 0.1% captopril gel (group II); and (iii) instilled 0.5% captopril gel intraurethrally (group III). RESULTS: In phase I, no significant local side-effects were seen in the urethra of rabbits. In phase II, the mean age of the patients was 39.5 and the mean follow-up duration was 16 months. The most common etiology of the urethral stricture in the patients was iatrogenic (35.7%), most of their strictures had a depth of 0.5 cm or less (67.8%), and the length of most strictures was between 1 and 2 cm (41.1%). The patients' maximum urine flow increased more in groups II and III, than in group I (P < 0.04, P < 0.05, respectively). The recurrence rate was less in groups II and III than in group I (P < 0.05). In terms of the maximal urine flow and recurrence rate, no significant difference was seen between group II and group III (P = 0.13, P = 0.21, respectively). CONCLUSION: Captopril gel is a safe, effective and non-toxic agent for decreasing the recurrence rate of the urethral stricture after internal urethrotomy. However, more studies, including more cases and a longer follow up, are needed to prove the effect of captopril gel on patients' urethra.

Left renal vein transposition for treatment of the Nutcracker syndrome.

The nutcracker syndrome refers to compression of the left renal vein between the aorta and the superior mesenteric artery, which results in renal vein and left gonadal vein varices. Herein, we report our experience with left renal vein transposition for treatment of the nutcracker syndrome. To the best of our knowledge, this is the first reported case of the left renal vein transposition for treatment of the nutcracker syndrome in Iran.

Open dismembered tubularized flap pyeloplasty: an effective and simple operation for treatment of ureteropelvic junction obstruction.

OBJECTIVE: To evaluate the success rate of dismembered tubularized flap pyeloplasty (DTFP) in the treatment of ureteropelvic junction obstruction (UPJO). PATIENTS AND METHODS: In a prospective study from August 2002 to September 2004, 15 patients with a mean age of 21 years (range 2-47) in whom UPJO had been diagnosed by sonography, excretory urography or diuretic renography and who had a large extrarenal pelvis, underwent operation via flank intercostal incision. The proximal ureter and renal pelvis were dissected and mobilized retroperitoneally, the site of UPJO was excised and the site of insertion of the ureter on the renal pelvis was closed with a stitch. A wide based renal pelvic flap was created and tubularized to bridge the upper ureteral defect. After insertion of a nephrostomy tube, a double-J tube was inserted as an internal ureteral stent and anastomosis of the tubularized flap to the spatulated upper ureter was done and the renal pelvis window was closed. Patients were followed 3, 6 and 12 months postoperatively. RESULTS: Mean operation time was 1 h and mean hospital stay was 3 days. The ureteral stent was removed 4 weeks after operation and at the same time a nephrostogram was done that showed a widely patent ureteropelvic junction with good renal pelvis drainage in 12 ( approximately 80%) of the cases, but in 3 cases (approximately 20%) passage of contrast materials was not seen. In these patients, methylene blue was injected via a nephrostomy tube and in 2 patients (14%) urine color turned blue 20 min later, but in 1 patient (7%) this test was also negative. The latter patient underwent percutaneous endopyelotomy later. Mean patient follow-up was 14 months. Follow-up excretory urography confirmed patent and unobstructed ureteropelvic junction in all patients. The overall success rate of DTFP was 93%. CONCLUSION: DTFP is a simple and effective procedure for patients with UPJO who have long or multiple upper ureteral strictures and a large extrarenal pelvis.

Can thymic tissue induce tolerance to kidney allografts?

INTRODUCTION: The aim of this study was to investigate the beneficial effect of donor thymic tissue to induce tolerance in thymokidney allografts, transplanted to thymectomized cross-bred canines. MATERIALS AND METHODS: Seven pairs of transplant donors and recipients were selected from 3- to 4-month-old cross-bred canines with major histocompatibility complex (MHC) mismatches. Recipients underwent partial thymectomy 4 weeks before transplantation and received an autologous thymic graft under the renal capsule, which had been engrafted in the donors 3 months before transplantation (thymokidney). Successful engraftment with evidence of thymocyte development in the donors was determined by gross and histologic examination at the time of transplantation. Biopsy specimens were obtained at the transplant day and 3 months after transplantation and were studied histologically for evidence of hyperacute or acute rejection. RESULTS: At 90 days after the operation, all 7 juvenile thymic grafts had developed with normal thymic structure under the renal capsule. Hyperacute rejection was not observed in allografts, and all of them were functioning until the end of follow-up; however, all of the allografts showed acute cell-mediated rejection 3 months after transplantation. CONCLUSION: No tolerance was induced by vascularized donor thymokidneys in MHC-mismatched canines. The advantages of tolerance over chronic immunosuppression are so great that a potentially tolerogenic approach such as thymic transplantation would seem worthy of further investigations on large animal models. To evaluate the beneficial effects of thymic tissue in tolerance induction, utilizing a short course, low-dose adjuvant immunosuppressant to this regimen and/or application of in-bred MHC-matched canines is suggested.

Primary realignment of posterior urethral rupture.

INTRODUCTION: We report the results of treatment of posterior urethral rupture (PUR) by primary realignment with some modifications of the technique. MATERIALS AND METHODS: In this prospective study, 25 patients (mean age, 33.5 years; range, 18 to 70 years) in whom PUR had been proved underwent primary urethral realignment. All patients were evaluated postoperatively for urinary incontinence, erectile dysfunction, and urethral stricture. They were followed for a mean of 20 months (range, 9 to 27 months). RESULTS: In 20 of 25 patients (80%), posterior urethral rupture was associated with pelvic fractures and in 2 (8%), bladder rupture was also present. None of the patients had urinary incontinence. Six patients (24%) had evidence of postoperative stricture that required urethral dilatation and/or direct vision internal urethrotomy in 2 or 3 procedures under local anesthesia. Erectile dysfunction was reported by 4 patients (16%) as a decreased quality of erection, all of whom responded to sildenafil. CONCLUSION: We believe that primary realignment of PUR is a simple procedure associated with low morbidity. It is recommended for patients who are stable and have no other significant intra-abdominal and pelvic organ injuries.

Apoptosis in transitional cell carcinoma of bladder and its relation to proliferation and expression of p53 and bcl-2.

Transitional cell carcinoma of bladder (TCC) is a relatively common cancer among men. Tumor progression is associated with expression or modulation of several gene products that control apoptosis and proliferation. Apoptosis is a negative growth regulatory mechanism in tumors. The aim of this study is to examine apoptosis and related regulatory molecular markers in a group of patients with TCC. Paraffinembedded tissues from 49 patients with TCC were examined for the expression of bcl-2, p53 and Ki-67 by immunohistochemistry. Apoptosis was detected by TUNEL method. Correlation between apoptotic index (AI), proliferation index (PI) and bcl-2 and p53 expression with each other and with pathological grade was determined. Apoptosis was observed in 28.1% of TCC cases. The mean AI of all cases was 13.7+/-24. No correlation was found between apoptosis and differentiation status of carcinoma. Bcl-2 expression was weakly detected in only one sample. P53 expression was detected in 26 of cases with mean staining index of 102+/-96. A significant correlation between p53 and Ki-67 staining indices was observed (r=0.521, p=0.001). Both p53 and Ki-67 expression showed a good association with the pathological grade (p=0.0001 and p=0.004, respectively). None of the markers showed significant correlation with AI and no correlation was found between the ratio of AI to PI and other parameters either. In conclusion, the frequency of apoptosis in TCC of bladder appears not to be associated with tumor grade, and with bcl-2, p53 and Ki-67 expression.