[A Case of Localized Amyloidosis of the Right Ureter Treated Effectively by Occulusive Dressing Technique Therapy Using Dimethyl Sulfoxide : Long-Term Follow-Up of Eight Years].

The patient was a 56-year-old female. She was referred to our department for further examination of right hydronephrosis in 2010. Computed tomography (CT) showed right hydronephrosis, and retrograde pyelography (RP) revealed stenosis of the right lower ureter. Urine cytology was negative. Transurethral biopsy of the right ureter was performed using ureteroscopic cup forceps and the histopathlogical diagnosis was ureteral amyloidosis. A whole-body search was performed, including rectal biopsy, but no evidence of amyloidosis was obtained. She was diagnosed with localized amyloidosis of the right ureter. A ureteral stent was indwelled and the patient was given occulusive dressing technique (ODT) therapy using dimethyl sulfoxide (DMSO) for 1 year. After ODT therapy, right hydronephrosis improved. After a 2-year followup, it worsened. ODT therapy was restarted and continued for 2 years. She consulted our department because of fever and right lumbago in April 2017 after a 4-month interruption of ODT therapy. CT revealed progression of the right hydronephrosis. A ureteral stent was indwelled and ODT therapy was restarted. The right hydronephrosis improved after 1 year. ODT therapy using DMSO was effective for localized ureteral amyloidosis, but periodic follow-up was necessary and ODT therapy was also effective when it recurred after the interruption of treatment.

[THREE CASES OF IATROGENIC URETERAL INJURY ASSOCIATED WITH TOTAL LAPAROSCOPIC HYSTERECTOMY].

We report three cases of iatrogenic ureteral injury associated with total laparoscopic hysterectomy. Case 1 was a 50-year-old woman. She underwent total laparoscopic hysterectomy (TLH) for myoma uteri. Postoperatively, a blood test revealed renal dysfunction, and ultrasonography (US) revealed left hydronephrosis. She was referred to our department on the next day of TLH. Computed tomography (CT) revealed left hydronephrosis. Retrograde pyelography (RP) was tried, but the ureteral catheter could not be inserted into the left ureteral orifice. An operation was performed on the second day after THL. The left ureter was ligated near the ureterovesical junction, and so uretero-ureterostomy was conducted. Case 2 was a 38-year-old woman. She underwent TLH for myoma uteri. Postoperatively, she complained of abdominal fullness and diarrhea. A blood test revealed renal dysfunction and US revealed left hydronephrosis on the ninth day after THL. She was referred to our department next day. CT revealed left hydronephrosis and ascitis. RP revealed extravasation of contrast medium from the left ureter. She was diagnosed with left ureteral injury accompanied by extravasation of urine into the intraperitoneal space. Uretero-cystoneostomy was performed. Case 3 was a 45-year-old woman. She underwent TLH for myoma uteri. Postoperatively, a blood test revealed renal dysfunction, and US revealed left hydronephrosis. She was referred to our department on the fifth day after TLH. CT revealed left hdronephrosis and ureteral obstruction of the left lower ureter. An operation was performed under laparoscopy, cystoscopy, and fluoroscopy on that day and the left lower ureter was kinked by threads. Cutting of the threads was performed under laparoscopy. After thread cutting, the kink of the ureter was improved and ureteral stent could be indwelled between the renal pelvis and bladder. After 1 month, the ureteral stent was removed and left hydronephrosis disappeared.

[THREE CASES OF TRANSURETHRAL RESECTION OF THE BLADDER TUMOR (TURBT), PERFORMED FOR BLADDER TUMOR ON THE ANTERIOR WALL, WITH BLADDER RUPTURE OCCURRING AFTER DISCHARGE].

Three cases are reported of TURBT on the anterior wall, with bladder rupture occurring after discharge. Patient 1 was a 68-year-old man. He had macroscopic hematuria and he strained to void a bloody clot on the 10th day after TURBT. Subsequently, right lower abdominal pain occurred. Computed tomography (CT) revealed the extravasation of contrast medium into the prevesical space. He was diagnosed with extraperitoneal bladder rupture, and a urethral catheter was indwelled. Cancer invasion of muscle was diagnosed by pathological examination and total cystectomy was scheduled one and a half months later, but the prostate could not be resected due to hard tissue around the bladder neck. Patient 2 was an 82-year-old man and had a history of radiation therapy for a muscle invasive bladder tumor. He complained of pollakisuria two weeks after TURBT, and renal failure was detected on a blood test. CT revealed ascites, and a urethral catheter was indwelled. Ascites disappeared, but the urethral catheter deviated into the abdominal cavity based on repeated CT the next day, and he was diagnosed with intraperitoneal bladder rupture. Emergent surgery was performed, and the ruptured part was sutured with omentum covering and a cystostomy was created. Patient 3 was an 83-year-old man undergoing treatment for benign prostatic hypertrophy (BPH). He had received bladder instillation therapy of Bacillus Calmette-Guerin (BCG) ten months previously. When urinating 6 days after TURBT, lower abdominal pain developed. CT demonstrated retroperitoneal bladder rupture, and a urethral catheter was indwelled. The urethral catheter was removed 6 days later, but lower abdominal pain occurred again the next day. Thus, the urethral catheter was re-indwelled for a further two weeks.In TURBT on the anterior wall or dome, for the patients who had previously received radiation therapy to the pelvis, or intravesical instillation therapy of the BCG or accompanied by urinary disturbance, such as BPH, it is necessary to consider bladder rupture after discharge.

[TWO CASES OF PEDIATRIC DEEP RENAL INJURY AT A LOCAL HOSPITAL].

We report two cases of pediatric deep renal injury at a local hospital. Case 1 was a 13-year-old girl. She fell from a bicycle and hit her back in a gutter. She complained of left back pain. Computed tomography (CT) revealed left deep renal injury accompanied by peripancreatic hematoma. Emergent surgery was performed and the transected kidney was resected, but pancreatic injury was not noted. Case 2 was a 10-year-old girl. She slipped during walking on her way home from school and hit her back on a concrete block. She complained of left back pain and gross hematuria. CT revealed left deep renal injury. Interventional radiology (IVR) was performed, but arterial bleeding was not noted, and so conservative therapy was performed. Although pediatric deep renal injury might be treated conservatively in general, treatment of such cases should be performed ideally at a hospital with IVR available for general anesthesia, and radiologists on-call in the event of any sudden change in the patient's condition. However, pediatric patients with serious renal injury, including the above, who cannot be transported to an advanced treatment hospital, can be treated through cooperation between skilled interventional radiologists and surgeons even in local hospitals with limited facilities and manpower.

[A Case of Metastasis of Cholangiocarcinoma to the Bladder].

The patient was an 88-year-old male. He was referred to the Department of Internal Medicine because of total body itching and jaundice in July 2011. The serum bilirubin level was elevated, and the serum CA19-9 level was also elevated to 266. 6 U/ml. Computed tomography (CT) and endoscopic retrograde cholangiopancreatography (ERCP) revealed a solid tumor between the hepatic hilus and common bile duct, and choler cytodiagnosis was class V ; adenocarcinoma. The patient was diagnosed with hilar cholangiocarcinoma and received conservative treatment with endoscopic nasobiliary drainage (ENBD) due to his advanced age. The patient was then referred to our department because CT revealed right hydronephrosis and thickening of the right side of the bladder wall, which had not been detected on admission in October 2011. Cystoscopy revealed a broad-based edematous tumor on the right side of the bladder. Transurethral resection of the bladder tumor (TURBT) was performed. The histological diagnosis was moderately differentiated tubular adenocarcinoma. Immunohistostaining using CA19-9 was performed, and cancer cells were positive. The final histology led to a diagnosis of metastasis of cholangiocarcinoma to the bladder. The patient died of liver failure in March 2012.

[FOUR CASES OF THE PROSTATIC ABSCESS IN WHICH DRAINAGE WAS EFFECTIVE].

We encountered four prostatic abscess patients. Although antimicrobial therapies were ineffective, drainage was effective in all cases. Patient 1 had lung cancer and diabetes mellitus (DM), and patient 2 developed acute prostatitis after transrectal prostatic biopsy. Culture of the urine and blood revealed extended-spectrum beta lactamase (ESBL) -producing Escherichia coli (E.coli). Patient 3 had previously sustained spinal cord injury, and urinated by self-catheterization. Patient 4 had untreated, severe DM. Patient 1, 2 and 3 had been treated by transurethral resection of the prostate (TURP). Patient 2 complained of ejaculatory incompetence after the surgery, and the symptom caused mental distress. Patient 4 was a 43-year-old man who had undergone transperineal needle aspiration under ultrasound guidance to avoid ejaculatory incompetence. The prostatic abscess disappeared in all cases after drainage without recurrence.

A Simple Method for Constructing Artificial Promoters Activated in Response to Ultrasound Stimulation.

It has been pointed out that ultrasound could be used as a controller for bioprocesses including gene expression since its energy can noninvasively reach deep in the body. Gene expression may be timely and spatially controlled by ultrasound, thus providing necessary bioactive proteins for the targeted tissue in a timely fashion. Although there are many processes involved in gene expression control, one of the most important processes is transcription, and the promoter plays an essential role in it. There are several promoters known to be activated in response to ultrasound irradiation . However, in our opinion, an artificial promoter is more suitable for clinical use. We herein describe simple methods to construct promoters that are responsive to ultrasound irradiation by randomly combining cis-elements (transcription factor binding motifs) and thereby improve its reactivity to ultrasound irradiation .

Analysis of repeated 24-core saturation prostate biopsy: Inverse association between asymptomatic histological inflammation and prostate cancer detection.

Saturation prostate biopsy protocols have been developed to improve the prostate cancer (PCa) detection rate, particularly in the setting of repeat biopsies. The present study attempted to clarify the association between PCa detection and various risk factors in repeat saturation biopsies. A retrospective analysis was conducted on 78 Japanese patients for whom findings had caused suspicion of PCa despite previous negative prostate biopsies, and who consecutively underwent a 24-core transperineal repeat biopsy at Toyama University Hospital (Toyama, Japan). PCa was confirmed histologically in 16 of the 78 patients (20.5%). A univariate analysis revealed that the prostate-specific antigen (PSA) level at repeat biopsy was higher (P<0.01), the fPSA/tPSA ratio was lower (P=0.04), the total prostate volume was smaller (P=0.01) and the PSA density was higher (P<0.01) in PCa patients than in patients with benign prostatic disease (BPD). Histological inflammation was more frequently observed in BPD patients than in PCa patients (P<0.01). A multivariate analysis revealed that histological inflammation was the only independent predictor of the presence of a malignant lesion on repeat biopsy (odds ratio, 0.027; P=0.01). It must be considered that inflammation may cause a PSA increase in some patients with a negative initial biopsy, leading to unnecessary repeat biopsy.

A CASE OF ADVANCED NON-SEMINOMATOUS TESTICULAR GERM CELL TUMOR ACCOMPANIED PANIC DISORDER.

We report a 33-year-old male with a left advanced non-seminomatous testicular germ cell tumor (NSGCT) accompanied panic disorder. He had experienced palpitation and hyperpnea in crowds in his twenties. He was admitted to the Department of Otorhinolaryngology with the chief complaint of left neck swelling. 18F-fluorodeoxy glucose positron emission tomography/computed tomography (FDG-PET/CT) demonstrated left neck, left supraclavicular, left axillary, and paraaortic lymph node (LN) swelling and left testicular swelling. He was referred to our department. The left testis had enlarged to the size of a fist. He rejected admission at that time, but next day, he was taken to our hospital by an ambulance because he lost consciousness at home. No abnormalities were found in the brain CT and electrocardiogram. He was admitted and left high orchiectomy was performed. The human chorionic gonadotropin (HCG) level had elevated to 9,717 IU/L and alpha fetoprotein level (AFP) had elevated to 427 ng/ml. The histopathological diagnosis was tumors of more than one histological type, mixed forms: seminoma and embryonal carcinoma.He had palpitation and hyperpnea after admission and was diagnosed with panic disorder by a psychiatrist. Psychotropic drugs (fluvoxamine maleate 50 mg/day, alprazolam 0.8 mg/day) were prescribed and the panic attacks disappeared afterwards. The psychiatric social worker supported his mind side. Bleomycin, etoposide, and cisplatin (BEP) therapy was performed for 4 courses. He put on a blanket to his face and came to avoid a conversation with other people during the chemotherapy. He was diagnosed with depression and psychotropic drugs were increased (fluvoxamine maleate 50→75 mg/day, alprazolam 0.8→1.2 mg/day) in quantity.Lymphadenectomies for LN metastases were performed and their histopathological examination revealed the existence of viable embryonal carcinoma in the supraclavicular LN. Etoposide, ifosfamide, and cisplatin (VIP) therapy was performed for 2 courses.The pateint has remained alive without tumor recurrence. Psychotropic drugs were reduced and the recent drug is fluvoxamine maleate 25 mg/day.

A CASE OF MULTIPLE VISCERAL INJURIES ACCOMPANIED BY RIGHT RENAL PEDICLE INJURY AND LEFT URETERAL DISRUPTION TREATED SUCCESSFULLY BY LEFT URETEROCALICOSTOMY.

We report a 25-year-old male with multiple visceral injuries accompanied by right renal pedicle injury and left ureteral disruption treated successfully by left ureterocalicostomy. He was accidentally crushed by a roller for fishing net hoists while working as a fisherman in May 2011. He was emergently transported to Kurobe City Hospital. He was in shock, but recovered with fluid therapy. CT revealed bilateral hemothorax, liver injury, bowel injury, right renal pedicle injury, left renal injury, and inferior vena cava damage. After bilateral chest drainage, emergent surgery was performed. Laparotomy revealed pancreatic injury, liver injury, disruption of the stomach and jejunum, colonic injury, and retroperitoneal hematoma on the right side. Distal pancreatectomy, hepatorrhaphy, left half resection of the colon, subtotal gastrectomy, and colostomy were performed. However, the bleeding of the right lobe of the liver could not be stopped, and gauze packing on the liver surface was performed for damage control. During the operation, right renal pedicle injury was not treated because the pulsation of the retroperitoneal hematoma was not palpable and the hematoma did not enlarge to the left side across the center; furthermore, his general condition was very poor. After the operation, the patient showed anuria, and hemodialysis was performed twice a week. One week after the operation, removal of the gauze was performed under general anesthesia. The gauze was removed from the liver while sprinkling physiological saline, and there was little bleeding. Tachocomb® (CSL Behring, Tokyo, Japan) was placed on the surface of the liver and a drainage tube was indwelled. Twenty-four days postoperatively, CT revealed left hydronephrosis with right nonfunctioning kidney, and percutaneus left nephrostomy was performed. Antegrade and retrograde pyelograms revealed a left ureteral defect of 8 cm in the upper ureter.The patient was introduced to the Department of Urology of Shinshu University Hospital. Left ureterocalicostomy was performed in January 2012, and the nephrostomy catheter was removed. The temporal colostomy was closed in the Department of Surgery of Kurobe City Hospital in May 2014. He subsequently resumed his normal life.

Ultrasound up-regulates expression of heme oxygenase-1 gene in endothelial cells.

PURPOSE: We found heme oxygenase-1 (HO-1) gene up-regulation in response to ultrasound stimulation in endothelial cells. This project was started to explore the mechanism since it could be applied clinically. METHODS: Quantitative real-time PCR experiments were conducted to determine the appropriate condition to up-regulate HO-1 gene expression. A promoter assay experiment and a microarray analysis were done to analyze signal transduction pathways to be activated in endothelial cells by ultrasound stimulation. RESULTS: Approximately, 33-fold up-regulation of HO-1 mRNA was observed when endothelial cells were exposed to 1-MHz ultrasound at 0.3 W/cm(2) and DF 10 % for 20 s. Data obtained from promoter analysis and microarray analysis suggested that the Nrf2/StRE signal pathway was involved in the up-regulation. In addition, ultrasound stimulation to endothelial cells changed its gene expression profile so that it could modify the anti-inflammatory, antioxidant, and vasodilatory activities in addition to pro-angiogenic and vascular reconstitution activities of endothelial cells. CONCLUSION: It was suggested that ultrasound stimulation of endothelial cells up-regulated HO-1 expression through intracellular oxidative stress, which subsequently changed the gene expression pattern so that it could modify vascular functions.

The acquired radioresistance in HeLa cells under conditions mimicking hypoxia was attenuated by a decreased expression of HIF subunit genes induced by RNA interference.

The cancer cells residing in the hypoxic layer are resistant to radiation and these are ones responsible for cancer recurrence after radiation therapy. One of the reasons why hypoxic cancer cells acquire radioresistance may be attributable to changes in the gene expression profile by the activation of hypoxia inducible factors (HIFs). However, the details underlying this process remain unknown. In this study, we investigated the effects of knockdown of HIF subunit genes to elucidate how HIF subunit genes may be involved in the radioresistance acquired by HeLa cells following exposure to a hypoxia mimic. Interestingly, HIF-1α and HIF-2α seemed mutually complementary for each other when either of them was suppressed. We thus suppressed the expression of both genes simultaneously. To do this, we developed a short hairpin RNA (shRNA) targeting a high homology region between HIF-1α and HIF-2α. It was shown that the expression of the shRNA effectively suppressed the acquisition of radioresistance following the hypoxia mimic. Moreover, it was confirmed that suppression of both subunits resulted in the downregulation of stem cell markers and the suppression of spheroid formation during the hypoxia mimicking-conditions. This shRNA-mediated knockdown method targeting a common region shared by a family of genes may offer a new candidate cancer treatment.

The effect of selective renal parenchymal clamping during laparoscopic partial nephrectomy on early postoperative renal function: a preliminary report.

PURPOSE: A major concern when performing laparoscopic partial nephrectomy (LPN) is potential postoperative renal dysfunction. The objective of this study was to compare the effects of LPN with selective renal parenchymal clamping (SRPC) (LPNSRPC) and LPN using microwave tissue coagulation (MTC) (LPNMTC) on postoperative renal function. MATERIALS AND METHODS: This study included 12 patients (5 men and 7 women) who underwent LPNSRPC (n = 6) or LPNMTC (n = 6) for exophytic tumors. Renal scanning with technetium-99m diethylenetriaminepentaacetic acid (Tc-99m DTPA) was performed preoperatively and postoperatively at 1 month in all patients. RESULTS: The mean tumor size, surgical duration, and intraoperative blood loss were similar in both groups. In the LPNMTC group, although not significant, the mean postoperative glomerular filtration rate (GFR) values in the affected kidneys were decreased compared to the preoperative values. When evaluating the affected renal function by split function (SF), the mean postoperative SF in the affected kidneys was significantly decreased compared to the preoperative value. In the LPNSRPC group, the mean postoperative GFR and SF in the affected kidneys were not significantly changed compared with the preoperative values. CONCLUSION: Our preliminary experience demonstrates that LPNSRPC facilitates maximal nephron-sparing surgery without collateral thermal damage causing renal impairment.

Multiple organ retraction during urological laparoscopic surgery.

PURPOSE: During laparoscopic surgery, as in open surgery, exposure is critical. We describe our surgical technique that provides intra-abdominal multiple organ retraction in order to obtain a stable surgical field and adequate exposure. MATERIALS AND METHODS: The Flex Arm™ surgical holder (Mediflex Surgical Products, Islandia, NY) is an adjustable, spring-loaded articulating instrument holder. The device offers the flexibility of the attachment of multiple arms (up to three) on the same rod, which can rotate 360° to aid in positioning. This device permits multiple laparoscopic instruments to be held simultaneously. We used this device for multiple organ retraction during urological laparoscopic surgery (retroperitoneoscopic radical prostatectomy, n=5; laparoscopic radical nephrectomy, n=3; laparoscopic partial nephrectomy, n=2; laparoscopic pyeloplasty, n=2). RESULTS: We achieved significant improvements in the efficiency of retraction of the liver, kidney, colon, and prostate during urological laparoscopic surgery. Our method maintains a constant surgical field through the proper retractor position and the proper amount of isometric force for optimal exposure. CONCLUSIONS: With this device, instrument migration due to muscle fatigue and organ injury as a result of excessive force are all substantially reduced. We believe that this technique may be helpful not only in urological laparoscopic surgery, but also in laparoscopic approaches in other departments.

Utilization of microRNAs with decreased expression levels in response to X-ray irradiation for fine-tuning radiation-controlled gene regulation.

We previously developed a promoter that was responsive to radiation by randomly combining cis-elements of transcription factors that are activated in response to radiation in prostate cancer cells. The promoter enhanced the expression of the luciferase gene linked downstream by more than 10-fold 12 h after X-ray irradiation at 10 Gy. However, without radiation, it still significantly drove its expression. To suppress expression while retaining its enhancement in response to radiation, we focused our attention on microRNAs (miRNAs). miRNAs are a group of non-coding RNAs approximately 22 nucleotides long that control gene expression by binding to a target sequence residing on the 3'-untranslated region (3'UTR) of a target gene. We identified 8 miRNAs that were downregulated in response to X-ray irradiation, and inserted artificial target sequences composed of randomly combined complementary sequences into 3 representative miRNAs into the 3'UTR of the luciferase gene. The target sequences suppressed the expression, and released the expression, after X-ray irradiation, as expected. When we combined an artificial target sequence with the radiation-responsive promoter, it resulted in a clear-cut gene regulation of expression that was greater than that induced by the promoter alone.

Laparoscopic resection of adrenal and extra-adrenal pheochromocytoma.

BACKGROUND AND PURPOSE: Laparoscopic resection of extra-adrenal pheochromocytoma (EAPs) necessitates meticulous surgical procedures because of changes in anatomic disposition and/or proximity to major blood vessels. Complete resection can be traumatic and may cause an increase in catecholamine levels. We present our experiences with laparoscopic resection of EAP (LEAP) and compare the intraoperative hemodynamics with those during laparoscopic resection of adrenal pheochromocytoma (LAP). PATIENTS AND METHODS: We retrospectively reviewed the medical records of five patients who underwent LEAP (retrocaval EAP, n=2; interaortocaval EAP, n=1; periadrenal EAP, n=2) and five who underwent LAP between October 2001 and October 2011. We also evaluated fluctuations in blood pressure (BP) reported during both surgeries. RESULTS: The tumors were successfully resected under laparoscopic guidance in both groups, and conversion to open surgery or blood transfusion was not needed. Intraoperative hypertension (BP>200 mm Hg) was observed in three LEAP and four LAP patients, whereas intraoperative hypotension (BP<80 mm Hg) was observed in five LEAP and three LAP patients. No significant differences were observed between groups, however. CONCLUSIONS: Laparoscopy is the method of choice for surgeons experienced in EAP resection because it is feasible and reproducible with appropriate preoperative planning, similar to LAP.

Expression of Hepatocyte growth factor activator inhibitor type-1 (HAI-1) in prostate cancer.

BACKGROUND: Hepatocyte growth factor activator inhibitor type-1 (HAI-1) inhibits hepatocyte growth factor activator and matriptase. In the present study it was investigated whether the expression of HAI-1 is associated with the progression of prostate cancer. PATIENTS AND METHODS: The expression of HAI-1 was evaluated by immunohistochemistry (IHC) of samples from 51 patients with negative prostate biopsies and 75 patients with untreated prostate cancer. Furthermore, the expression of HAI-1 was evaluated in 24 patients with castration-resistant prostate cancer (CRPC), and the relationship between HAI-1 expression and the prostate-specific antigen (PSA) progression-free rate was investigated. RESULTS: Expression of HAI-1 by IHC in patients with prostate cancer was significantly higher than in those with negative prostate biopsy. CRPC exhibited significantly lower HAI-1 expression than untreated metastatic prostate cancer. The PSA progression-free rate was worse in patients without HAI-1 expression than in those with positive HAI-1 expression. CONCLUSION: It is suggested that HAI-1 may play an important role in the pathogenesis of CRPC.

Early quality of life outcomes in patients with prostate cancer managed by high-dose-rate brachytherapy as monotherapy.

OBJECTIVES: To evaluate the early quality of life outcomes in prostate cancer patients managed by high-dose-rate brachytherapy as monotherapy. METHODS: A total of 51 patients with cT1c-T3aN0M0 prostate cancer treated between July 2007 and January 2010 were included in this study. The average age was 69 years, and the average initial serum prostate-specific antigen was 10.98 ng/mL. A total of 25, 18 and eight patients were considered to be low, intermediate and high risk, respectively. All patients received one implant of Ir-192 and seven fractions of 6.5 Gy within 3.5 days for a total prescribed dose of 45.5 Gy. For high-risk prostate cancer, neoadjuvant androgen deprivation therapy was carried out for at least 6 months, and continued after high-dose-rate brachytherapy. Quality of life outcomes were measured by using the International Prostate Symptom Score, the Functional Assessment of Cancer Therapy-Prostate and the International Index of Erectile Function Questionnaire. The oncological outcome was assessed by serum prostate-specific antigen and diagnostic imaging. Adverse events were also recorded. RESULTS: The Functional Assessment of Cancer Therapy-Prostate scores decreased for a few months after high-dose-rate brachytherapy, and recovered to pretreatment condition thereafter. The International Prostate Symptom Score significantly increased 2 weeks after treatment for each of its items and their sum, and it returned to baseline after 12 weeks. Sexual function decreased at 2 and 4 weeks, and recovered after 12 weeks. Severe complications were rare. Within a median follow up of 17.2 months, two patients showed a prostate-specific antigen recurrence. CONCLUSIONS: High-dose-rate brachytherapy for prostate cancer is a feasible treatment modality with acceptable toxicity and only a limited impact on the quality of life.

Regulation of gene expression in human prostate cancer cells with artificially constructed promoters that are activated in response to ultrasound stimulation.

We chose promoters responsive to sonication in LNCap cells, a prostate cancer cell line, out of a library composed of DNA fragments constructed by linking the TATA box sequence to randomly combined cis-acting elements of transcription factors activated in response to radiation in prostate cancer cells. When a plasmid containing the luciferase gene under control of a promoter was transfected into LNCap cells and sonicated with 1 MHz ultrasound at 0.5 W/cm(2), 10% DF for 60s, 13 promoters showed more than 10-fold enhancement compared with their counterparts without sonication 12h after sonication. As to their responsiveness to sonication, the best two promoters were then compared to clone 880-8, a derivative from clone 880 that was created by random introduction of point mutations and was shown to have an improved response to X-ray irradiation. We then took clone 880-8 for further analyses since it showed the highest enhancement to sonication, though not statistically significant from the others. Next, we employed a retrovirus vector and stably introduced the luciferase gene under control of clone 880-8 into LNCap cells to establish a cell line. When the cell line was sonicated with 1 MHz ultrasound at 0.5 W/cm(2), 10% DF for 60s, luciferase expression was enhanced up to 14.8-fold 12h after sonication. We then established another cell line by replacing the luciferase gene with the fcy::fur gene, a suicide gene, and when the cell line was sonicated with 1 MHz ultrasound at 0.5 W/cm(2), 10% DF for 60s, expression of the gene was enhanced, showing the maximum expression 12-24h after sonication. When the cells were incubated in medium containing 5-fluorocytosine, cell survival ratio decreased dose dependently with 5-fluorocytosine only after sonication treatment, suggesting this promoter could be utilized for gene expression control with ultrasound.

Development of a therapeutically important radiation induced promoter.

Radio-genetic therapy is a combination of radiation therapy and gene therapy that may solve some of the problems associated with conventional radiotherapy. A promoter responsive to radiation was obtained from a promoter library composed of DNA fragments created by linking the TATA box signal to randomly combined binding sequences of transcription factors that are reactive to radiation. Each promoter connected to the luciferase gene, was evaluated by luciferase expression enhancement in transfected cells after X-ray irradiation. The reactivity of the best promoter was improved by the random introduction of point mutations and the resultant promoter showed more than a 20-fold enhancement of the luciferase expression after X-ray irradiation at 10 Gy. The expression of downstream genes was also enhanced in stably transfected cells not only by X-rays but also by proton beam irradiation; and either enhancement was attenuated when an anti-oxidant was added, thus suggesting the involvement of oxidative stress in the promoter activation. Constructed promoters were also activated in tumors grown in mice. In addition, cell killing with the fcy::fur gene (a suicide gene converting 5-fluorocytosin to highly toxic 5-fluorouracil) increased dose-dependently with 5-fluorocytosin only after X-ray irradiation in vitro. These results suggest that promoters obtained through this method could be used for possible clinical applications.