Retroperitoneal robot-assisted laparoscopic upper pole heminephrectomy in adult patients with duplex kidneys.

Background: To report our experience with retroperitoneal robot-assisted laparoscopic upper pole heminephrectomy in adult patients with duplex kidneys. Methods: We retrospectively reviewed the medical records of 7 patients who underwent retroperitoneal robot-assisted laparoscopic upper pole heminephrectomy at our institution between September 2014 and July 2017. Of the robot-assisted laparoscopic procedures, 5 were on the left and 2 on the right side. Results: All patients underwent robot-assisted laparoscopic surgery successfully in a totally retroperitoneal manner without conversion to open surgery. The mean operative time was 175 mins (range 140-270). The mean estimated blood loss was 84 mL (range 20-200). The mean postoperative hospital stay was 7 days (range 5-9). No major intraoperative and postoperative complications occurred. All patients had a resolution of their presenting symptoms after surgery at a mean follow-up of 24 months (range 14-38). Conclusion: Our initial clinical experience suggests that robot-assisted laparoscopic upper pole heminephrectomy using a retroperitoneal approach for a duplex kidney appears to be safe with acceptable perioperative outcomes.

Study on Dual Channel Lateral Field Excitation Quartz Crystal Microbalance for Measuring Liquid Electrical Properties.

Lateral field excitation quartz crystal microbalance (LFE-QCM) can detect both the electrical properties (conductivity and permittivity) and mechanical properties (viscosity and density) of the liquid. In practical applications for detecting electrical properties, the viscosity and density of the liquid will also change. This research proposed a dual-channel LFE-QCM for reducing the influence of density and viscosity. The sensing layer of one resonant element is almost bare, and the other is covered by a metal film as a reference. Different organic solutions and NaCl solution were used to study the influence of mechanical properties and the temperature on electrical properties. The experimental results demonstrate that the dual-channel LFE-QCM is necessary for properly detecting electrical properties of the liquid.

Robotic-assisted Urethra-sparing Simple Prostatectomy Via an Extraperitoneal Approach.

OBJECTIVE: To present an original technique of robotic-assisted urethra-sparing simple prostatectomy (RAUSP) for treating patients with benign prostatic hyperplasia. MATERIALS AND METHODS: From April 2015 to December 2016, 27 patients underwent RAUSP via an extraperitoneal approach. Baseline patient characteristics, perioperative outcomes, pathologic outcomes, postoperative Clavien complications, International Prostate Symptom Score, International Index of Erectile Function, and ejaculatory function were assessed. RESULTS: Twenty-six patients (96.3%) successfully underwent RAUSP, one patient (3.7%) was converted to simple prostatectomy. Median operative time was 169 minutes (interquartile range: 150-185); median estimated blood loss was 235 mL (interquartile range: 180-300). Seven cases (26.9%) required urethral repair secondary to inadvertent urethrotomy. Mean catheterization time was 1.6 days (range 1-5). Clavien complications were reported, 6 being low grade (grade 1 or 2) with a single 3a complication (gross hematuria requiring bladder irrigation). Mean follow-up duration was 16.4 months (range 9-30). Postoperative questionnaire demonstrated that international prostate symptom score (P < .001) and quality of life score (P < .001) were significantly improved postoperatively. A total of 14 patients reported erectile function, 13 of which had normal ejaculation, only 1 complained retrograde ejaculation. CONCLUSION: RAUSP is technically feasible for patients with benign prostatic hyperplasia. Our data indicate that patients have short catheterization time, an acceptable risk profile, significant improvements of voiding function and maintaining antegrade ejaculation following this urethral-sparing technique.

Extraperitoneal robot-assisted laparoscopic retroperitoneal lymph node dissection for early-stage testicular nonseminomatous germ cell tumors: A case report and literature review.

RATIONALE: Typically robot-assisted laparoscopic retroperitoneal lymph node dissection (R-RPLND) has been performed via a transperitoneal approach. Herein we report the first case of a novel R-RPLND using an extraperitoneal approach. PATIENT CONCERNS: A 38-year-old man presented with an enlarging right scrotal mass. DIAGNOSES: Scrotal ultrasonography demonstrated a 5.5-cm solid mass of the right testis. The patient underwent right radical inguinal orchiectomy. Pathologic examination demonstrated a mixed germ cell tumor, predominately embryonal carcinoma with yolk sac tumor. INTERVENTIONS: Extraperitoneal R-PRLND was performed 3 weeks after the radical orchiectomy. OUTCOME: The final pathologic examination showed a count of 19 lymph nodes, all of them negative. Normal antegrade ejaculation returned within 4 weeks postoperatively. No retroperitoneal recurrence or elevation of tumor marker levels were seen via surveillance imaging. LESSONS: Our study shows that extraperitoneal R-RPLND is a safe and feasible procedure using an extraperitoneal approach that provides minimal invasion and rapid recovery of patients.

Focal adhesion kinases crucially regulate TGFß-induced migration and invasion of bladder cancer cells via Src kinase and E-cadherin.

Focal adhesion kinase (FAK) is a non-receptor protein-tyrosine kinase that is triggered off by special extracellular signals such as some growth factors and integrins. FAK is found in cell-matrix attachment sites and implicated in cell migration, invasion, movement, gene expression, survival and apoptosis. In this study, we aimed to investigate whether FAK plays a role in invasion and migration of bladder cancer cells. Using an FAK-specific small interfering RNA (siRNA) and an FAK inhibitor PF-228, we found that inhibition of FAK tyrosine phosphorylation or knockdown of FAK suppressed invasion and migration of bladder cancer cells. Src is an important mediator of FAK-regulated migratory and invasive activity. Tyrosine phosphorylation of Src and FAK is mutually dependent and plays a key role in transforming growth factor beta (TGFß)-induced invasion and migration. E-cadherin acts downstream of FAK and is a critical negative regulator in FAK-regulated invasion and migration of bladder cancer cells. These findings imply that FAK is involved in oncogenic signaling of invasion and migration, which can be a novel therapeutic target to treat patients with bladder cancer.

Castleman's Disease Presenting in the Pelvis.

Castleman's disease (CD) is a rare benign lymphoid disorder with unknown etiology. There are 2 major forms of this disease, unicentric (ie, localized CD) and multicentric, which play a major role in determining therapy. The mediastinum is the most common localization for the localized CD, whereas its occurrence in the pelvis is even rarer. We report the case of a 22-year-old woman who had a pelvic mass located in the region of the left iliac fossa. The patient subsequently underwent a robotic-assisted laparoscopic tumorectomy. Pathological examination revealed pelvic localized CD of the hyaline-vascular type. CD should be included in the differential diagnosis of pelvic masses that are noted in the pelvic cavity.

Inhibition of focal adhesion kinase induces apoptosis in bladder cancer cells via Src and the phosphatidylinositol 3-kinase/Akt pathway.

Focal adhesion kinase (FAK) is a 125-kDa, cytosolic, non-receptor, protein tyrosine kinase localized at focal adhesions that can be activated by multiple inputs and in different manners. FAK is implicated in signaling pathways regulating cell movement, invasion, survival, gene expression and cancer stem cell self-renewal. The aim of the present study was to investigate whether FAK plays a role in the apoptosis of bladder cancer cells. The study employed in situ deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling and Annexin V labeling flow cytometry. It was found that both the knockdown of FAK and the suppression of FAK phosphorylation were able to induce apoptosis in bladder cancer cells. Caspase-3 was activated during the apoptosis induced by the suppression of FAK phosphorylation. Src was involved in FAK-regulated apoptosis in bladder cancer cells, while the suppression of Src phosphorylation was able to inhibit FAK tyrosine phosphorylation and induce apoptosis. Furthermore, phosphatidylinositol 3-kinase (PI3K)/Akt signaling was inhibited via the suppression of FAK tyrosine phosphorylation. Conversely, the expression of neither the general nor the tyrosine-phosphorylated FAK was regulated by inhibiting PI3K/Akt, which suggested that PI3K/Akt acted downstream of FAK to regulate apoptosis in bladder cancer cells. These findings indicate the presence of a mechanism of apoptosis involving FAK-mediated oncogenic signaling. FAK may function as an important regulator of extracellular signaling-mediated apoptosis in bladder cancer and be used as a novel therapeutic target in the treatment of the condition.

Monitoring of prostate cancer growth and metastasis using a PSA luciferase report plasmid in a mouse model.

OBJECTIVE: To construct a PSA luciferase report plasmid and monitor the growth and metastasis of prostate cancer after emasculation in SCID mice. METHODS: PSA promoter sequence and luciferase gene were amplified by PCR and subsequently inserted into pZsGreen1-1 vector to construct pPSA-FL-Luc vector. LNCaP cells that were stably transfected with pPSA-FL-Luc were used to establish a SCID mouse xenograft model. Then, the growth and metastasis of prostate cancer were monitored via living imaging. RESULTS: We successfully constructed a PSA luciferase plasmid, pPSA-FL-Luc. DHT enhanced luciferase activity in a concentration-dependent manner in 293T cells with pPSA-FL-Luc transfection. Prostate cancer SCID mouse model was established with pPSA-FL-Luc transfected LNCaP cells. In tumor bearing mice with or without emasculation, pPSA-FL-Luc plasmid was applied to monitored tumor growth and metastasis based on bioluminescence imaging. CONCLUSIONS: We construct a pPSA-FL-Luc plasmid, which stably expresses luciferase and can be applied to monitor tumor development in a prostate SCID mouse model.

Ureteral obstruction and urinary fistula due to fibrin glue after partial nephrectomy: A case report and review of the literature.

In the present study, we report the case of a 69-year-old female who developed urinary leakage following partial nephrectomy (PN) to remove left renal masses. The results of CT and MR urography revealed left proximal ureteral obstruction and urinary fistula. Reoperation was performed on the 16th postoperative day to explore the left kidney and ureter in order to relieve the obstruction. The left proximal ureter was found to be enfolded by fibrin glue and showed marked stiffness and adhesion during the reoperation. The lesion of the ureter was resected and the ureter was anastomosed with the routine double-J stent. Pathological examination of surgical specimens revealed fat fibrous scar tissue hyperplasia with inflammatory cell infiltration. The patient recovered completely without exudate. Our experience suggests that care should be taken to avoid touching the ureter with fibrin glue during PN surgery.

Epigallocatechin-3-gallate inhibits bladder cancer cell invasion via suppression of NF-κB­mediated matrix metalloproteinase-9 expression.

Bladder cancer is the fourth most common cancer in males and the ninth most common in females. However, despite the numerous advances made in the past few decades, the prognosis of patients with bladder cancer remains poor. Metastasis is one of the major causes of mortality in bladder cancer patients. Therefore, the inhibition of metastasis is one of the most significant issues in bladder cancer research. The present study was conducted to evaluate the anti-metastatic potential of (-)-epigallocatechin gallate (EGCG, the major phytochemical in green tea) against bladder cancer and its mechanism of action. EGCG efficiently and dose-dependently inhibited adhesion, migration and invasion of T24 human bladder cancer cells. Mechanistically, EGCG inhibited phosphatidylinositol 3'-kinase/Akt activation that resulted in inactivation of nuclear factor-κB (NF-κB) and the inhibition of the expression of matrix metalloproteinase-9 (MMP-9), ultimately suppressing invasion and metastasis. These findings suggest that EGCG is a potential therapeutic candidate against tumor invasion.

Tea consumption and risk of bladder cancer: a meta-analysis.

BACKGROUND: Tea consumption has been reported to be associated with an decreased risk of several types of cancers. However, the results based on epidemiological studies on the association of tea consumption with bladder cancer were inconsistent. This meta-analysis was undertaken to evaluate the relationship between tea consumption and bladder cancer risk. METHODS: Eligible studies were retrieved via both computer searches and review of references. The summary relative risk (RR) with 95% confidence interval (CI) was calculated. RESULTS: Twenty three studies met the inclusion criteria of the meta-analysis. No association with bladder cancer was observed in either overall tea consumption group (OR =0.94, 95% CI 0.85-1.04) or subgroups stratified by sex, study design, geographical region or tea types. CONCLUSIONS: Our findings did not support that tea consumption was related to the decreased risk of bladder cancer.

The construction of an oxalate-degrading intestinal stem cell population in mice: a potential new treatment option for patients with calcium oxalate calculus.

About 80% of all urological stones are calcium oxalate, mainly caused by idiopathic hyperoxaluria (IH). The increased absorption of oxalate from the intestine is the major factor underlying IH. The continuous self-renewal of the intestinal epithelium is due to the vigorous proliferation and differentiation of intestinal stem cells. If the intestinal stem cell population can acquire the ability to metabolize calcium oxalate by means of oxc and frc transgenes, this will prove a promising new therapy option for IH. In our research, the oxalate-degrading genes of Oxalobacter formigenes (Oxf)-the frc gene and oxc gene-were cloned and transfected into a cultured mouse-derived intestinal SC population to give the latter an oxalate-degrading function. Oxf was isolated and cultivated and the oxalate-degrading genes-frc and oxc-were cloned. The dicistronic eukaryotic expression vector pIRES-oxc-frc was constructed and transferred into the mouse stem cell population. After selection with G418, the expression of the genes was identified. The oxalate-degrading function of transfected cells was determined by transfection into the intestinal stem cell population of the mouse. The change in oxalate concentration was determined with an ion chromatograph. The recombinant plasmid containing oxc and frc genes was transfected into the stem cell population of the mouse and the expression of the genes found normal. The cell population had acquired an oxalate-degrading function. The oxc and frc genes could be transfected into the intestinal stem cell population of the mouse and the cells acquired an oxalate-degrading function.

Selective targeting of HPV-16 E6/E7 in cervical cancer cells with a potent oncolytic adenovirus and its enhanced effect with radiotherapy in vitro and vivo.

Recent studies have shown that oncolytic adenovirus specifically targeted tumor cells while sparing normal cells. Here, we report a novel E1A-mutant adenovirus (M6) with antisense HPV16 E6 E7 DNA inserted into the deleted 6.7K/gp19K region of E3. The target effects of M6 on HPV16-positive cervical cancer cells were evaluated in vivo and in vitro. By using cytopathic effect (CPE) and viral replication assays, we verified M6 was competent to selectively replicate in cervical cancer cells in vitro. Moreover, we found infection of M6 was able to inhibit the expression of HPV16 E6 and E7 oncogenes and induce apoptosis of HPV16-positive cervical cancer cells. Further analysis in vitro revealed that the invasive ability of SiHa cells was significantly inhibited by M6. To determine if M6 synergized with radiotherapy-induced anti-tumor activity against HPV16-related cancer cells, we transfected SiHa cells with M6 followed by a single exposure to radiation. A significantly suppression of cell growth and induced apoptosis was observed in SiHa cells received M6 transfection combined with radiotherapy. Animal experiments showed that M6 transfection notably improved the survival of tumor-bearing mice in combination with radiotherapy, much superior to that of those treated by Adv5/dE1A plus radiation or M6 alone. These findings indicated the anti-tumoral efficacy of M6 on HPV16-positive cervical cancer cells and its synergic therapeutic application in radiation for cervical cancer.

Inhibition of TGF-beta receptor I by siRNA suppresses the motility and invasiveness of T24 bladder cancer cells via modulation of integrins and matrix metalloproteinase.

BACKGROUND: Urinary bladder transitional-cell carcinoma is still challenging because the mechanisms underlying the tumor progression are still largely unknown. Transforming growth factor beta1 (TGF-beta1) is considered a crucial molecule in the tumorigenesis of urinary bladder carcinoma. Many studies have indicated that it is also associated with epithelial-mesenchymal transition, angiogenesis, migration and metastases in many types of malignant tumors. MATERIALS AND METHODS: We blocked the TGF-beta signal pathway in T24 human bladder cancer cells with a siRNA (TsiRNA), which targets the TGF-beta type I receptor and evaluated the effects of TGF-beta1 and TsiRNA on the cell motility and invasiveness by Matrigel migration assay, wound-healing assay and Matrigel invasion assay. RT-PCR and Western blotting analysis were used to examine the effects of TGF-beta1 and TsiRNA on the expression of TGFBRI and genes, which are related to tumor migration and invasion. RESULTS: While exogenous TGF-beta1 enhanced the migration and invasion of T24 cells, TsiRNA significantly suppressed them. RT-PCR and Western blotting analysis revealed that TsiRNA could downregulate both the expression of alpha3, beta1 and alpha2 integrin subunits and the activity of matrix metalloproteinase 9 enhanced by exogenous TGF-beta1. CONCLUSION: Our study suggested that inhibition of TGF-beta1 signaling pathway by siRNA could be beneficial in the treatment of patients with metastatic bladder cancer.

[Construction of oxalate-degrading intestinal stem cell population in mice].

OBJECTIVE: The oxalate-degradation genes of Oxalobacter formigenes (Ox.F)-frc gene and oxc gene-were cloned and transfected into intestinal stem cell population of the mouse to make the latter obtain oxalate-degradation function. METHODS: (1) The dicistronic eukaryotic expression vector, which could express oxc gene and frc gene in the same time, pIRES-oxc-frc was constructed. (2) The intestinal stem cell population of the mouse were isolated and cultured, and the function of the cell growth and differentiation was identified. (3) The cells were transfected with pIRES-oxc-frc. After selection by G418, the function of the cell growth and differentiation and the the expression of the objective genes were identified. (4) The concentration of the oxalate in the culture medium which was used to culture the transgenic cells was determined by ion chromatography to explore the oxalate-degradation function of the cells. RESULTS: Ox.F could be isolated and cultured from the feces of Chinese people. Compared with the foreign reports, a certain morphologic variation of the Ox.F existed. But the oxc gene and frc gene showed high homology with the sequence reported in GenBank. The recombinant plasmid containing oxc gene and frc gene could successfully be transfected into the intestinal stem cell population of the mouse. The expression of the objective genes was normal. The concentration of the oxalate in the culture fluid of the transgenic intestinal stem cell population [(2.48 +/- 0.03 g/L)] was lower than those of the normal group [(2.69 +/- 0.01) g/L] and the control group [(2.69 +/- 0.01) g/L, P < 0.01]. CONCLUSION: The oxc gene and the frc gene could be transfected into the intestinal stem cell population of the mouse, and the cells could be given oxalate-degrading function. The gene of prokaryocyte could be introduced into the eukaryocyte for a successful expression.

Estrogen in combination with 5-azacitidine up-regulates p75NTR expression and induces apoptosis in 22Rv1 prostate cancer cells.

Previous studies suggest that the low-affinity neurotrophin receptor p75NTR inhibits the proliferation of human prostate cancer cells, and that estrogen interacts with p75NTR in many tissues. In this study, we exposed 22Rv1 androgen-independent prostate cancer cells to 17-ß-estradiol and the DNA demethylating agent 5-azacitidine (5-AzaC) to explore the interactions between estrogen and p75NTR. We found that estrogen induced estrogen receptor (ESR) subtype?2 and p75NTR expression in 22Rv1 cells, and that 5-AzaC further enhanced these effects. Estrogen in combination with 5-AzaC induced cell apoptosis, which was associated with the inhibition of NF-κB translocation to the nucleus. These results provide evidence that the up-regulation of ESR2 and p75NTR by estrogen plus 5-AzaC may be a potential therapeutic strategy for the treatment of androgen-refractory prostate cancer.

Up-regulation of E-cadherin by small activating RNA inhibits cell invasion and migration in 5637 human bladder cancer cells.

Recent studies have reported that chemically synthesized small duplex RNAs complementary to promoters of target genes can specifically induce gene expression in several cancer cell lines. Such dsRNA, referred to as small activating RNA (saRNA), are involved in the recently described phenomenon called RNA activation (RNAa). Recent findings show that saRNA can inhibit cell proliferation and viability via up-regulation of p21(WAF1/CIP1) (p21) in human bladder cancer cells. In the present study, we demonstrate that induction of E-cadherin expression by saRNA leads to suppression of migration and invasion of 5637 human bladder cancer cells in vitro. The elevated E-cadherin expression was confirmed at transcriptional and protein levels after transfection of a 21-nucleotide dsRNA targeting the E-cadherin promoter (dsEcad). Furthermore, this inhibitory effect was associated with relocalization of beta-catenin from the nucleus to the plasma membrane and decreased beta-catenin-mediated transactivation. These data suggest that activation of E-cadherin by saRNA may have a therapeutic benefit for bladder and other types of cancer.

RNA interference against HPV16 E7 oncogene leads to viral E6 and E7 suppression in cervical cancer cells and apoptosis via upregulation of Rb and p53.

The simultaneous expression of human papillomavirus type 16 (HPV16) E6 and E7 oncogenes is pivotal for malignant transformation and maintenance of malignant phenotypes. Silencing these oncogenes is considered to be applicable in molecular therapies of human cervical cancer. However, it remains to be determined whether HPV16 E6 and E7 could be both silenced to obtain most efficient antitumor activity by using RNA interference (RNAi) technology. Herein, we designed a small interfering RNA (siRNA) targeting HPV16-E7 region to degrade either E6, or truncated E6 (E6*) and E7 mRNAs and to simultaneously knockdown both E6 and E7 expression. Firstly, the sequence targeting HPV16-E7 region was inserted into the shRNA packing vector pSIREN-DNR, yielding pSIREN-16E7 to stably express corresponding shRNA. HPV16-transformed SiHa and CaSki cells were used as a model system; RT-PCR, Western Blotting, MTT assay, TUNEL staining, Annexin V apoptosis assay and flow cytometry were applied to examine the effects of pSIREN-16E7. Our results indicated that HPV16-E7 specific shRNA (16E7-shRNA) induced selective degradation of E6 and E7 mRNAs and proteins. E6 silencing induced accumulation of cellular p53 and p21. In contrast, E7 silencing induced hypophosphorylation of retinoblastoma (Rb) protein. The loss of E6 and E7 reduced cell growth and ultimately resulted in massive apoptotic cell death selectively in HPV-positive cancer cells, compared with the HPV-negative ones. We demonstrated that 16E7-shRNA can induce simultaneous E6 and E7 suppression and lead to striking apoptosis in HPV16-related cancer cells by activating cellular p53, p21 and Rb. Therefore, RNAi using E7 shRNA may have the gene-specific therapy potential for HPV16-related cancers.