Chang-Te Lin

Methylomics analysis identifies ZNF671 as an epigenetically repressed novel tumor suppressor and a potential non-invasive biomarker for the detection of urothelial carcinoma.

The molecular mechanism underlying the lethal phenomenon of urothelial carcinoma (UC) tumor recurrence remains unresolved. Here, by methylation microarray, we identified promoter methylation of the zinc-finger protein gene, ZNF671 in bladder UC tumor tissue samples, a finding that was independently validated by bisulphite pyrosequencing in cell lines and tissue samples. Subsequent assays including treatment with epigenetic depressive agents and in vitro methylation showed ZNF671 methylation to result in its transcriptional repression. ZNF671 re-expression in UC cell lines, via ectopic expression, inhibited tumor growth and invasion, in possible conjunction with downregulation of cancer stem cell markers (c-KIT, NANOG, OCT4). Clinically, high ZNF671 methylation in UC tumor tissues (n=96; 63 bladder, 33 upper urinary tract) associated with tumor grade and poor locoregional disease-free survival. Quantitative MSP analysis in a training (n=97) and test (n=61) sets of voided urine samples from bladder UC patients revealed a sensitivity and specificity of 42%-48% and 89%-92.8%, respectively, for UC cancer detection. Moreover, combining DNA methylation of ZNF671 and 2 other genes (IRF8 and sFRP1) further increased the sensitivity to 96.2%, suggesting a possible three-gene UC biomarker. In summary, ZNF671, an epigenetically silenced novel tumor suppressor, represents a potential predictor for UC relapse and non-invasive biomarker that could assist in UC clinical decision-making.

Tubeless Percutaneous Nephrolithotomy for Geriatric Patients

Aim: To evaluate the safety of tubeless percutaneous nephrolithotomy (PCNL) in geriatric patients. Materials and Method: This is a retrospective review of 401 patients who received tubeless PCNL in a single institute. Among these, 50 were performed in patients aged older than 70 years (group 1), while 351 were performed in the remaining younger patients (group 2). Results: There was no significant difference in stone size between the 2 groups (3.6 ± 1.9 vs. 3.5 ± 2.0 cm). The average operative time was similar in both groups (92.8 ± 34.5 vs. 86.6 ± 32.0 min). The stone-free rate in groups 1 and 2 was 68.0% (34/50) and 83.8% (294/351), respectively, which was statistically significantly different. The average postoperative hospital stay was longer in group 1 (4.6 ± 3.4 days) than in group 2 (3.9 ± 2.5 days), but the difference was not statistically significant. There was no significant difference in postoperative urinary tract infection rate and blood transfusion rate in both groups (urinary tract infection: 18.0 vs. 8.8%; blood transfusion: 4 vs. 2.6%). Two patients in group 1 and 3 patients in group 2 experienced pulmonary complications. There was no other severe complication. Conclusion: Tubeless PCNL is a safe procedure for the treatment of geriatric patients with urolithiasis.

The high incidence of JC virus infection in urothelial carcinoma tissue in Taiwan.

Human polyomaviruses, JC virus (JCV) and BK virus (BKV), usually remain latent in kidney and urothelial tissue after primary infection. Infection with human polyomavirus has still not been correlated conclusively with malignancy of kidney and urothelial tissue. The present study investigated further the possible relationship between JCV/BKV infection and urothelial carcinoma. Tissue samples were examined from 33 urothelial carcinomas and 5 renal cell carcinomas for JCV/BKV infection, using nested PCR with primers common to both JCV and BKV. The viral genotypes were further verified by endonuclease digestion and DNA sequencing following the PCR. In addition, immunohistochemistry and Western blotting were also performed to detect viral large tumor protein (LT) and the late capsid protein (VP1) in the tissue samples. The results from nested PCR showed that 90.1% (30/33) of the urothelial carcinomas samples and all of the renal cell carcinomas samples (5/5) were JCV DNA positive. Both archetypal and re‐arranged JCV genotypes were detected. On the other hand, BKV DNA was detected in only one (3%) of the urothelial carcinoma tissue samples. The immunohistochemical results showed that 30% (10/33) of urothelial carcinoma tissues was stained positive for large tumor antigen (LT). However, the structural protein VP1 was not detectable in any of the tissue samples examined. The present study demonstrated that JCV is highly prevalent in urothelial carcinoma tissue as is the expression of large tumor antigen. Therefore, the findings support the hypothesis that JCV infection is associated with urothelial carcinoma. J. Med. Virol. 83:2191–2199, 2011.

Prognostic relevance of preoperative circulating CD8-positive lymphocytes in the urinary bladder recurrence of urothelial carcinoma

OBJECTIVE Tumor-infiltrate lymphocytes (TIL) have been associated with favorable outcomes in various tumors including urothelial carcinoma (UC). There is little literature about peripheral blood lymphocytes (PBL). Our objective is to investigate the clinical significance and relevance of PBL on the outcomes of UC patients. MATERIALS AND METHODS Ninety UC treated patients at Chia-Yi Christian hospital were enrolled. Preoperative PBLs were collected and analyzed for the percentage of each subpopulation of lymphocyte using flow cytometry. The prognostic values were calculated by using Kaplan-Meier curve and Cox progression model for univariate and multivariate analyses, respectively. Furthermore, available tumor specimens from 27 patients were further analyzed for number of CD8(+) tumor infiltrating lymphocytes (TIL) using immunohistochemistry. The correlation between percentage of CD8+ PBL and number of CD8+ TIL was analyzed using a linear regression model. RESULTS The log-rank test showed that tumor location (urinary bladder vs. upper urinary tract), enrolled status (primary or recurrent), and CD8(+) PBL were significant prognostic indicators of recurrence (P values, 0.043, 0.039, and 0.018, respectively). Cox analyses showed that CD8(+) PBL was the sole independent prognostic indicator for recurrence-free survival (P = 0.048). The results using a linear regression analysis showed there was a reverse correlation between CD8(+) TIL and PBL (r(2) = 0.635, P < 0.0001). CONCLUSIONS In our investigation, preoperative CD8(+) PBL was an independent predictor for bladder recurrence. The percentages of CD8(+) PBL were reversely correlated with the number of TIL. Such findings may benefit in the decision for subsequent intravesical therapy after surgery.

Bilateral ureteral complete obstruction with huge spontaneous urinoma formation in a patient with advanced bladder cancer

Spontaneous rupture of the collecting system with extravasation of urine and urinoma formation is usually associated with urinary tract obstruction by a ureteral calculus. Tumor growth is an extremely rare cause of urinary extravasation. Here we report a case of bilateral obstructive uropathy with a huge spontaneous left retroperitoneal urinoma caused by advanced infiltrative transitional cell carcinoma of the urinary bladder. The point of leakage was located in the left renal pelvis. The urinary leakage ceased after percutaneous nephrostomy drainage, and the patient subsequently underwent radical cystoprostatectomy. Histopathology revealed a high-grade urothelial carcinoma of the urinary bladder with pelvic lymph node metastasis. The patient refused any adjuvant treatment and expired 6 months after the operation from disseminated metastasis from bladder cancer.

Dysfunction of natural killer cells in patients with transitional cell carcinoma

Previous studies had established that transforming growth factor-beta1 (TGF-beta1) is highly expressed in bladder tumor, facilitating progression and spreading of the cancerous cells. Here, we report that both the number and the cytotoxic function of natural killer (NK) cells are decreased in patients with superficial transitional cell carcinoma (TCC). In consistent with previously reported findings, the plasma TGF-beta1 concentration is also elevated in patients with superficial TCC. In vitro, the cytotoxic function of NK cells was impaired by the presence of TGF-beta1. Hence, our data suggested elevated concentration of serum TGF-beta1 in loss of NK cytotoxicity in superficial TCC patients, implicating altered innate immunity may correlate with the incidence of TCC.

Hyperbaric oxygen therapy for cyclophosphamide-induced intractable refractory hemorrhagic cystitis in a systemic lupus erythematosus patient.

Hemorrhagic cystitis is a complication of systemic lupus erythematosus and is also a common side effect after cyclophosphamide therapy. Intractable hemorrhagic cystitis is not unusual and may be a life-threatening condition; it has no effective noninvasive treatment at present. We report a case of hemorrhagic cystitis with intractable refractory bleeding that occurred in a 40-year-old woman after cyclophosphamide treatment for systemic lupus erythematosus. The hemorrhage was resistant to various therapies but resolved after hyperbaric oxygen therapy. There was no recurrent hematuria after hyperbaric oxygen therapy during 6 months of follow-up.

Delayed Pneumothorax After Tubeless Percutaneous Nephrolithotomy

Percutaneous nephrolithotomy is the treatment of choice for large urinary stone and staghorn stone. Supracostal access through the upper calyx provides a straight tract along the long axis of the kidney and is the optimal route for the treatment of staghorn stone. However, the supracostal access bears higher risk for pleural or lung injury resulting in hydrothorax or pneumothorax. Percutaneous nephrolithotomy induced pneumothorax usually occurs immediately after operation. We report a case of delayed pneumothorax after tubeless percutaneous nephrolithotomy for a complete staghorn stone.