RIN1 promotes renal cell carcinoma malignancy by activating EGFR signaling through Rab25.

We previously identified the important role of RIN1 expression in the prognosis of clear cell renal cell carcinoma (ccRCC). The role of RIN1 in ccRCC malignancy and underlying molecular mechanisms remain unclear. Here we report that ccRCC cells and tissues expressed more RIN1 than normal controls. Gain-of-function and loss-of-function studies demonstrated that RIN1 enhanced ccRCC cell growth, migration and invasion abilities in vitro and promoted tumor growth and metastasis in vivo. Mechanistic studies revealed that RIN1 has an activating effect on EGFR signaling in ccRCC. In addition, we unveil Rab25, a critical GTPase in ccRCC malignancy, as a functional RIN1 interacting partner. Knockdown of Rab25 eliminated the augmentation of carcinoma cell proliferation, migration and invasion by ectopic RIN1. We also confirmed that RIN1 and Rab25 expression correlates with the overall-survival of ccRCC patients from TCGA. These findings suggest that RIN1 plays an important oncogenic role in ccRCC malignancy by activation of EGFR signaling through interacting with Rab25, and RIN1 could be employed as an effective therapeutic target for ccRCC.

Validation of DAB2IP methylation and its relative significance in predicting outcome in renal cell carcinoma.

We have recently reported tumor suppressive role of DAB2IP in RCC development. In this study, We identified one CpG methylation biomarker (DAB2IP CpG1) located UTSS of DAB2IP that was associated with poor overall survival in a cohort of 318 ccRCC patients from the Cancer Genome Atlas (TCGA). We further validated the prognostic accuracy of DAB2IP CpG methylation by pyrosequencing quantitative methylation assay in 224 ccRCC patients from multiple Chinese centers (MCHC set), and 239 patients from University of Texas Southwestern Medical Center at Dallas (UTSW set) by using FFPE samples. DAB2IP CpG1 can predict the overall survival of patients in TCGA, MCHC, and UTSW sets independent of patient age, Fuhrman grade and TNM stage (all p<0.05). DAB2IP CpG1 successfully categorized patients into high-risk and low-risk groups with significant differences of clinical outcome in respective clinical subsets, regardless of age, sex, grade, stage, or race (HR: 1.63-7.83; all p<0.05). The detection of DAB2IP CpG1 methylation was minimally affected by ITH in ccRCC. DAB2IP mRNA expression was regulated by DNA methylation in vitro. DAB2IP CpG1 methylation is a practical and repeatable biomarker for ccRCC, which can provide prognostic value that complements the current staging system.

A CpG-methylation-based assay to predict survival in clear cell renal cell carcinoma.

Clear cell renal cell carcinomas (ccRCCs) display divergent clinical behaviours. Molecular markers might improve risk stratification of ccRCC. Here we use, based on genome-wide CpG methylation profiling, a LASSO model to develop a five-CpG-based assay for ccRCC prognosis that can be used with formalin-fixed paraffin-embedded specimens. The five-CpG-based classifier was validated in three independent sets from China, United States and the Cancer Genome Atlas data set. The classifier predicts the overall survival of ccRCC patients (hazard ratio=2.96-4.82; P=3.9 × 10(-6)-2.2 × 10(-9)), independent of standard clinical prognostic factors. The five-CpG-based classifier successfully categorizes patients into high-risk and low-risk groups, with significant differences of clinical outcome in respective clinical stages and individual 'stage, size, grade and necrosis' scores. Moreover, methylation at the five CpGs correlates with expression of five genes: PITX1, FOXE3, TWF2, EHBP1L1 and RIN1. Our five-CpG-based classifier is a practical and reliable prognostic tool for ccRCC that can add prognostic value to the staging system.

Transurethral bipolar plasma kinetic resection of ejaculatory duct for treatment of ejaculatory duct obstruction.

To evaluate the efficacy of transurethral bipolar plasma kinetic resection of ejaculatory duct for ejaculatory duct obstruction. The clinical information of 42 cases of ejaculatory duct obstruction was analyzed between July 2008 and June 2012. The diagnostic criteria included semen analysis, fructose and neutral α-glucosidase measurement in seminal plasma, transrectal ultrasonography, magnetic resonance imaging and vasography necessarily. Endoscopic procedure with bipolar plasma kinetic resection of ejaculatory duct was performed in all patients. Among these cases followed up 6 ≈ 24 months after operation, 38 patients (90.5%) had improved semen parameters, 23 azoospermic patients (60.5%) had sperm in the semen and 13 patients' wife (31%) achieved pregnancies in 42 cases of bipolar plasma kinetic resection of ejaculatory duct. Postoperative complications ensued as epididymitis in 1 case, watery ejaculate in 1, but no serious complication was observed. Bipolar plasma kinetic resection of ejaculatory duct appears to represent a promising endoscopic treatment alternative for ejaculatory duct obstruction patients, with high efficacy, less complications, quicker recovery and satisfactory follow-up parameters.

A modified single-armed technique for microsurgical vasoepididymostomy.

This study is to evaluate the effectiveness of a modified single-armed suture technique for microsurgical vasoepididymostomy (VE) in patients with epididymal obstructive azoospermia. From September 2011 to December 2011, microsurgical two-suture longitudinal intussusception VEs were performed using our modified single-armed suture technique in 17 men with epididymal obstructive azoospermia at our hospital. Two of these patients underwent repeated VEs after previous failed VEs, and one patient underwent unilateral VE because of an occlusion of the left abdominal vas deferens. The presence of sperm in the semen sample at 3 months postoperation was used as the preliminary endpoint of this study. Each patient provided at least one semen sample at the 3-month time point, and the patency was assessed by the reappearance of sperm (>10(4) ml(-1)) in the semen. The mean operative time for the modified technique was 219 min. Patency was noted in 10 men (58.8%), including one patient who underwent repeated VE. The patient who underwent unilateral anastomosis manifested no sperm postoperatively in his semen. Sperm granulomas were not detected in this cohort. The results of this study demonstrate that our modified technique for microsurgical longitudinal intussusception VE is effective. We believe that it is a practical alternative that may reduce operation time and obviate the suture crossing.

Microsurgical denervation of the spermatic cord for treatment of idiopathic chronic orchialgia.

A patient referred to our hospital, diagnosed with left idiopathic chronic orchialgia, was evaluated with a thorough medical and psychiatric history, physical examination, scrotal ultrasound and magnetic resonance imaging. Conservative management failed. The patient had temporary pain relief after undergoing outpatient cord block three times. Microsurgical denervation of the left spermatic cord was operated in March, 2011. A pain questionnaire was used to determine efficacy before and after operation, and complete pain relief was noted at one week after operation. The follow up period was 12 months, at the end of which the pain score was still zero. No complications, including testicular atrophy and hydrocele, occurred. Microsurgical denervation of the spermatic cord can be a minimally invasive, safe and effective management option for treatment of idiopathic chronic orchialgia.

Prostate-specific membrane antigen can promote in vivo osseous metastasis of prostate cancer cells in mice

Reports remain insufficient on whether and how prostate-specific membrane antigen (PSMA) can influence in vivo osseous metastasis of prostate cancer (PCa). In the present study, the authors induced stable expression of PSMA in mouse PCa cell line RM-1. In vivo osseous metastasis was induced in 37 6-week-old female C57BL/6 mice weighing 22.45 ± 0.456 g. RM-1 cells were actively injected into the femoral bone cavity, leading to bilateral dissymmetry of bone density in the femoral bone. Tumor cells were also detected in bone tissue by pathological examination. The impact on bone density was demonstrated by the significant difference between animals injected with RM-PSMA cells (0.0738 ± 0.0185 g/cm²) and animals injected with RM-empty plasmid cells (0.0895 ± 0.0241 g/cm²). The lytic bone lesion of the RM-PSMA group (68.4%) was higher than that of the control group (27.8%). Immunohistochemistry showed that the expression of both vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) was distinctly higher in the RM-PSMA group than in the control group, while ELISA and Western blot assay indicated that VEGF and MMP-9 were higher in the RM-PSMA group compared to the control group (in vitro). Thus, the present study proposed and then confirmed for the first time that PSMA can promote in vivo osseous metastasis of PCa by increasing sclerotic destruction of PCa cells. Further analyses also suggested that PSMA functions positively on the invasive ability of RM-1 by increasing the expression of MMP-9 and VEGF by osseous metastases in vivo.

Prostate-specific membrane antigen can promote in vivo osseous metastasis of prostate cancer cells in mice.

Reports remain insufficient on whether and how prostate-specific membrane antigen (PSMA) can influence in vivo osseous metastasis of prostate cancer (PCa). In the present study, the authors induced stable expression of PSMA in mouse PCa cell line RM-1. In vivo osseous metastasis was induced in 37 6-week-old female C57BL/6 mice weighing 22.45 ± 0.456 g. RM-1 cells were actively injected into the femoral bone cavity, leading to bilateral dissymmetry of bone density in the femoral bone. Tumor cells were also detected in bone tissue by pathological examination. The impact on bone density was demonstrated by the significant difference between animals injected with RM-PSMA cells (0.0738 ± 0.0185 g/cm²) and animals injected with RM-empty plasmid cells (0.0895 ± 0.0241 g/cm²). The lytic bone lesion of the RM-PSMA group (68.4%) was higher than that of the control group (27.8%). Immunohistochemistry showed that the expression of both vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) was distinctly higher in the RM-PSMA group than in the control group, while ELISA and Western blot assay indicated that VEGF and MMP-9 were higher in the RM-PSMA group compared to the control group (in vitro). Thus, the present study proposed and then confirmed for the first time that PSMA can promote in vivo osseous metastasis of PCa by increasing sclerotic destruction of PCa cells. Further analyses also suggested that PSMA functions positively on the invasive ability of RM-1 by increasing the expression of MMP-9 and VEGF by osseous metastases in vivo.

[Efficacy of transurethral resection of ejaculatory duct for treatment of ejaculatory duct obstruction: report of 60 cases].

OBJECTIVE: To evaluate the effect of transurethral resection of ejaculatory duct (TURED) for treatment of ejaculatory duct obstruction (EDO). METHODS: The clinical data of 60 cases of EDO from Oct. 2004 to Oct. 2010 were analyzed. The diagnostic criteria included semen analyses, fructose in seminal plasma, transrectal ultrasonography (TRUS), and vasography if necessary. All the patients were treated by TURED. Post-operative semen assay, postoperative patency rate and postoperative impregnation rate were followed. RESULTS: Semen analyses in the majority of cases showed the typical characteristics of EDO, azoospermia, low semen volume (average 1.10 ± 0.76 mL), low pH (average 6.5 ± 1.4), absent or low semen fructose (average 5.86 ± 2.19 µmol/one ejaculation). TRUS showed pure dilation of both ejaculatory ducts in 15 cases, prostatic cyst in 6, pure dilated seminal vesicles on both sides in 10, unilateral dilated seminal vesicle in 4, dilation of both ejaculatory duct and seminal vesicles in 10, dilated seminal vesicles with prostatic cyst in 5, unilateral dilated seminal vesicle and contralateral aplasia of seminal vesicle in 2, dilated seminal vesicles with dilation and calcifications of both ejaculatory ducts in 3. The remaining 5 had unilateral dilated ejaculatory duct and seminal vesicle with contralateral aplasia of seminal vesicle. In all the cases followed up more than 6 to 78 months after TURED, 51 patients (85.0%) had improved semen parameters and 16 patients' wives (26.7%) had pregnancies. CONCLUSION: TURED may be the simple, minimally invasive and effective method for the treatment of EDO.

Was fine-needle vasography an obsolete diagnostic method to evaluate ejaculatory duct obstruction? Report of 37 cases.

OBJECTIVES: Based on comparative analyses of the precision as well as reliability of the diagnostic results on fine-needle vasography (FNV) and transrectal ultrasonography (TRUS) in the diagnosis of patients who were suspected of having ejaculatory duct obstruction (EDO), this paper aims to demonstrate whether or not FNV techniques still constitute the backbone of diagnostic imaging used in obstructive azoospermia of EDO. MATERIALS AND METHODS: From July 2005 to December 2008, opening fine-needle punctuation and vasography were performed under local anesthesia in 37 infertile male patients in our center, all of whom were suspected of having bilateral EDO. The diagnostic criteria of these infertile males included characteristic changes of TRUS and/or characteristic changes of semen analysis. Comparative analyses were made of the diagnostic results of the two methods. RESULTS: In the 37 male patients, 19 (51.35%) had been confirmed with pathognomonic findings of both semen analyses and TRUS on complete EDO. Of the others, 18 patients (48.65%) testified with only pathognomonic findings of TRUS but not the typical characteristics on semen analyses. When compared with diagnostic results via FNV however, there were only 5 patients (13.51%) who could be confirmed as having bilateral EDO, 16/37 showed obstruction of the epididymis and proximal vas deferens, 7/37 showed bilateral vasal multiple obstruction, 2 showed unilateral EDO and multiple obstruction of the contralateral vas deferens, and the other 6 patients showed unilateral vasal multiple obstruction and obstruction on the contralateral epididymis and proximal vas deferens. Free flow of the contrast medium could be defined into the bladder by vasography in 23 patients. Of the 23 patients, 12 were suspected of having delayed evacuation based on more than 50% contrast medium remaining inside their seminal vesicles after exhaustion of urine. CONCLUSION: Our analyses suggest that despite its advantages of being non-invasive, the diagnostic effect of TRUS is limited by both accuracy and reliability. In contrast, although an invasive radiographic imaging method, FNV has been proven as being informative when providing preoperative details about obstruction and other symptoms. After 3 years of research we came to the conclusion that opening fine-needle punctuation and vasography have not become outdated, at least recently, and will remain a necessary and effective method to identify diseases and to provide a strictly defined morphology of the seminal vesicle and ductal system for more comprehensive diagnosis of obstructive azoospermia of EDO.

[Surgical treatment of obstructive azoospermia: a report of 56 cases].

OBJECTIVE: To evaluate the diagnosis and surgical treatment of obstructive azoospermia. METHODS: We analyzed the clinical data of 56 cases of obstructive azoospermia, 43 of them with ejaculatory duct obstruction (EDO), and the other 13 suspected of epididymal obstruction. The diagnostic methods included semen analyses, measurement of fructose and neutral alpha-glucosidase in the seminal plasma, transrectal ultrasonography (TRUS), and vasography when necessary. The 43 patients with EDO were treated by transurethral resection of the ejaculatory duct (TURED), and 11 of the 13 cases of suspected epididymal obstruction were confirmed by scrotal exploration and underwent either bilateral or unilateral vasoepididymostomy. The patients were followed up for 3 -51 months for postoperative semen quality and impregnation. RESULTS: Of the 43 azoospermia patients with EDO treated by TURED, 36 (83.7%) showed improved semen parameters and 11 (25.6%) achieved pregnancies. Among the 11 cases of azoospermia with confirmed epididymal obstruction treated by vasoepididymostomy, 6 (54.5%) had sperm in the semen assay and 3 (27.3%) achieved pregnancies. CONCLUSION: Semen analyses, measurement of fructose and neutral alpha-glucosidase in the seminal plasma, TRUS and vasography are important diagnostic methods for obstructive azoospermia. TURED is effective for azoospermia with EDO, while vasoepididymostomy is preferable for the treatment of azoospermia with epididymal obstruction.

[Diagnosis and treatment of iatrogenic ejaculatory duct obstruction following prostatic hyperthermia (report of 3 cases)].

OBJECTIVE: To investigate the characteristics of diagnosis and treatment of iatrogenic ejaculatory duct obstruction following prostatic hyperthermia. METHODS: Retrospective analyses were made of the clinical data of 3 cases of ejaculatory duct obstruction following prostatic hyperthermia. RESULTS: The 3 cases were diagnosed as urethrostenosis and deformity of the posterior urethra by transurethral ultrasound and semen analysis. And all the 3 patients had undergone prostatic hyperthermia for prostatitis. Transurethral resection of the ejaculatory duct (TURED) was performed to remove the obstruction and the postoperative semen analysis showed both semen volume and sperm count to be normal. CONCLUSION: Urethra microwave thermotherapy, urethra radiofrequency, or per urethra rheophore ablation can be adopted in the treatment of protatitis, but should be strictly indicated and cautiously selected lest secondary iatrogenic ejaculatory duct obstruction should result. For the treatment of this obstruction, TURED is the first choice.