Urine miR-21-5p as a potential biomarker for predicting effectiveness of tadalafil in benign prostatic hyperplasia.

AIM: To investigate whether urine levels of miRNAs that regulate the function of endothelial cells are associated with effectiveness in benign prostatic hyperplasia (BPH) patients treated with a phosphodiesterase type 5 inhibitor, tadalafil. PATIENTS & METHODS: We measured urine levels of three miRNAs (miR-21-5p, miR-126-5p & miR-155-5p) in 55 BPH patients before and after tadalafil administration to understand its effectiveness. RESULTS: Baseline urine miR-21-5p level was an independent predictor of response to tadalafil in multivariate regression analysis (odds ratio: 0.28; 95% CI: 0.10-0.77; p = 0.014). Receiver operator curve analysis revealed that baseline urine miR-21-5p could serve as a predictor of response (area under curve: 0.85; 95% CI: 0.75-0.95; p < 0.001). CONCLUSION: Urine miR-21-5p could serve as a biomarker in predicting response of tadalafil for BPH.

Preoperative granulocyte-colony stimulating factor (G-CSF) treatment improves congested liver regeneration.

BACKGROUND: Hepatic venous congestion after liver surgery can cause liver failure. We evaluated the effectiveness of granulocyte-colony stimulating factor (G-CSF) for regeneration of remnant liver with venous congestion. MATERIALS AND METHODS: Rats were divided into three groups. Group A underwent 60% hepatectomy. Group B underwent 60% hepatectomy with partial venous congestion in remnant liver. Group C underwent the same procedures as group B with G-CSF given preoperatively for 5 d. To evaluate liver regeneration in each group at 1, 2, 3, 5, and 7 d postoperatively, the proliferating cell nuclear antigen (PCNA) labeling index (LI), mitotic index (MI) in the congested area of remnant liver, and regeneration rate of remnant liver weight were measured. CD34 antibody-positive hematopoietic stem cell (HSC) colonies were identified using immunopathological staining. RESULTS: Compared with group B, in group C, LI, and MI in congested remnant liver increased earlier, peak LI value in the congested area appeared 24h earlier, and full recovery of remnant liver weight occurred 2 d earlier. On postoperative day 3, remnant liver weight was significantly greater in group C than group B. On microscopy, CD34-positive cells were seen in preoperative group C livers. CONCLUSION: Preoperative G-CSF improves regeneration of livers with venous congestion.

Reduction of fibrosis in a rat model of non-alcoholic steatohepatitis cirrhosis by human HGF gene transfection using electroporation.

BACKGROUND AND AIM: To study the histological changes caused by transfection of the hepatocyte growth factor (HGF) gene using electroporation (EP) in a non-alcoholic steatohepatitis (NASH) cirrhotic liver model. METHODS: NASH cirrhotic livers were prepared by administering a choline-deficient diet to 5-week-old male Wister rats for 12 weeks. Three groups of rats were used: rats in the G(+) group were transfected with the GFP gene using EP, rats in the H(+) group were transfected with the HGF gene using EP, and rats in the H(-) group were only injected with the HGF gene. Rats were sacrificed 2 days after gene transfection, and the Azan positive rate (APR) and Sudan positive rate (SPR) were calculated to evaluate fibrosis and fatty changes. RESULTS: The APR of the NASH cirrhotic livers was significantly higher than that in the normal livers. The APR did not decrease in the G(+) group and the H(-) group, but decreased significantly in the nonelectroporated as well as electroporated areas of the H(+) group. For SPR, there were no significant differences between the G(+), H(-), and H(+) groups. CONCLUSION: The improvement of fibrosis was not significant when a direct injection of the HGF gene was used alone, but it was enhanced by the concomitant use of EP. However, no efficacy was observed in fat components. These findings suggest that transfection of the HGF gene by EP may lead to an improvement of irreversible cirrhotic livers to reversible fatty livers.

Improvement of the survival rate after rat massive hepatectomy due to the reduction of apoptosis by caspase inhibitor.

BACKGROUND AND AIM: Acute liver failure after massive hepatectomy is caused by both necrosis and apoptosis in the remnant liver. We investigate the protective effect of the caspase inhibitor on apoptosis after massive hepatectomy in rats. METHODS: Benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone (ZVAD-fmk) is a general inhibitor of the caspase. Male Wister rats weighing 200-300 g were divided into three groups: 90Hx group undergoing 90% hepatectomy, 95Hx group undergoing 95% hepatectomy, 95Hx + ZVAD group undergoing 95% hepatectomy and administration of ZVAD-fmk. The 7-day survival rate was studied, and the rats were sacrificed at the 1, 2, 3, 5, and 7th day after hepatectomy. The remnant liver tissues were stained with hematoxylin-eosin, and with proliferating cell nuclear antigen (PCNA) for evaluation of liver regeneration, and with TdT-mediated dUTP-biotin nick end labeling (TUNEL) and in situ oligo ligation method (ISOL) for evaluation of apoptosis. RESULTS: The 7-day survival rates were 100%, 0%, and 30%, in the 90Hx, 95Hx, and 95Hx + ZVAD groups, respectively. There was no significant difference in PCNA labeling index (LI) between the 95Hx and 95Hx + ZVAD groups. TUNEL and ISOL LI of 95Hx + ZVAD group were significantly lower than those of 95Hx group. Fatal liver failure after massive hepatectomy was characterized by more apoptosis and less mitosis of hepatocytes. ZVAD-fmk could significantly attenuate apoptosis of hepatocytes in the remnant liver and improve the survival rate after 95% hepatectomy in rats. CONCLUSION: Caspase inhibitors such as ZVAD-fmk may provide a new adjuvant therapy to treat liver failure after massive hepatectomy.

Study on the relation of the shape of the uroflowmetrogram and the urethral loss coefficient calculated from the uroflowmetrogram.

The shape of the uroflowmetrogram reflects voiding conditions. Using a voiding simulation, we examined whether the urethral loss coefficient (LC) calculated from the approximated uroflowmetrogram correlates with parameters that regulate the shape of the uroflowmetrogram. A total of 161 normal and abnormal uroflowmetrograms were used. Normal female subjects and patients before and after transurethral resection of the prostate (TURP) were also studied. The ratio of maximum flow rate (Q(max)) to flow time (T), a parameter expressing the shape of the uroflowmetrogram, was calculated. The uroflowmetrograms were approximated using a voiding model, and the urethral LC was calculated. As a result, a strong negative correlation was observed between the Q(max)-flow time ratio, Q(max)/ T, and LC. Q(max)/T is the vertical to horizontal ratio of the uroflowmetrogram and indicates the average degree of acceleration of flow rate during voiding. On the other hand, urethral LC, which can be estimated from the shape of the uroflowmetrogram, is considered a kind of urethral resistance. We concluded that when urethral resistance is high, the degree of acceleration of flow rate is low on average. Our study also indicated that Qmax/T was less affected by voided volume (VV) compared to Q(max). As Q(max)/T is not as dependent on VV, it is useful for comparing cases with different VV.

Comparative antitumor activity of 5-fluorouracil and 5'-deoxy-5-fluorouridine in combination with interferon-alpha in renal cell carcinoma cell lines.

OBJECTIVES: Although interferon-alpha (IFNalpha) and interleukin-2 are used in the treatment of advanced renal cell carcinoma (RCC), the rate of efficacy is about 15% and not satisfactory. Therefore, a more effective treatment is being investigated. In this study, we examined the combined effects of IFNalpha and 5-fluorouracil (5-FU) as well as 5-deoxy-5-fluorouridine (5'-DFUR), a produrg of 5-FU, in vitro. MATERIALS AND METHODS: Four RCC cell lines (OCUU1, OCUU3, OCUU4, OCUU5) were established at our laboratory from RCC patients. OS-RC-2, RCC10RGB, TUHR14TKB, TUHR4TKB, A498 and Caki-1 were obtained from a commercial source. The sensitivity of the 10 RCC cell lines to 5-FU and 5'-DFUR was evaluated using MTT assay. The IC50 value for the cytostatics was expressed as the concentration at which growth was inhibited by 50% as compared with the control value. Thymidine phosphorylase (TP), the enzyme that converts 5'-DFUR into 5-FU and 5-FU into FdUMP, was estimated by ELISA. RESULTS: When the 10 RCC cell lines were divided into the low TP expression group and high TP expression group at 2.0 U/ml protein, TP expression was not enhanced by IFNalpha in all 4 cell lines in the low TP expression group. Antitumor effects were not enhanced by IFNalpha in 3 out of 4 cell lines for 5-FU and in all 4 cell lines for 5'-DFUR. On the other hand, in the high TP expression group, TP expression was enhanced by IFNalpha in 5 out of 6 cell lines, and antitumor effects were enhanced by IFNalpha in 5 out of 6 cell lines for 5-FU and in all 6 cell lines for 5'-DFUR. In addition, there was a significant correlation between TP expression and sensitivity to 5-FU and 5'-DFUR in all RCC cell lines. CONCLUSIONS: These results suggested that TP may be useful as a predictive factor in combination therapy with IFNalpha and 5-FU or 5'-DFUR, which may be a promising treatment for advanced RCC.

Chondrosarcoma of the urinary bladder and establishment of a human chondrosarcoma cell line (OCUU-6).

Chondrosarcoma is a very rare tumor of the urinary bladder, with only 4 cases reported to date. In this study, we report on a case of a 73-year-old male who presented bladder mass and right hydroureteronephrosis. Radical cystectomy, right nephrectomy and left ureterocutaneoustomy were performed, and histological study disclosed chondrosarcoma of the urinary bladder. As reported in other cases, the tumor was highly aggressive with a short clinical course, and the patient died of carcinomatous pleuritis at one month after surgery. Subsequently, we successfully established a human chondrosarcoma cell line (OCUU-6) from the pleural effusion of the patient.

Chondrosarcoma of the urinary bladder and establishment of a human chondrosarcoma cell line (OCUU-6).

Chondrosarcoma is a very rare tumor of the urinary bladder, with only 4 cases reported to date. In this study, we report on a case of a 73-year-old male who presented bladder mass and right hydroureteronephrosis. Radical cystectomy, right nephrectomy and left ureterocutaneoustomy were performed, and histological study disclosed chondrosarcoma of the urinary bladder. As reported in other cases, the tumor was highly aggressive with a short clinical course, and the patient died of carcinomatous pleuritis at one month after surgery. Subsequently, we successfully established a human chondrosarcoma cell line (OCUU-6) from the pleural effusion of the patient.

Role of tumor-associated macrophages in renal cell carcinoma.

We investigated the biologic meaning of tumor-associated macrophages (TAM) in renal cell carcinoma (RCC). The study group comprised of 83 patients with RCC. TAM was isolated by plastic adherence following enzymatic digestion of surgically removed tumor tissues. In some of the patients, monocytes were also isolated from peripheral blood mononuclear cells by plastic adherence. When TAM and monocytes were compared in the same patients, TAM produced interleukin 6 (IL-6), tumor necrosis factor alpha (TNFalpha) and interleukin 1beta (IL-1beta) without lipopolysaccharide (LPS) stimulation, while monocytes hardly produced IL-6, TNFalpha and IL-1beta without LPS stimulation. However, with LPS stimulation, monocytes produced more IL-6, TNFalpha and IL-1beta than TAM. In stage T1 RCC patients, there was a significant positive correlation between TNFalpha production of TAM and tumor size. In order to investigate the effects of TAM on cancer cells, TAM was co-cultured with A498, K562 and in some cases, with short-term established RCC lines for 96 h. As a result, TAM largely enhanced cell proliferation. These results suggested that TAM may play an important role in certain steps of tumor progression.

Effects of tumor necrosis factor alpha in renal cell carcinoma.

Tumor necrosis factor alpha (TNFalpha) and TNFalpha receptor mRNA expression, the effects of TNFalpha on DNA synthesis and cell proliferation as well as its effects on interleukin-6 (IL-6) production and expression in renal cell carcinoma (RCC) were studied using RCC cell lines. The effects of TNFalpha on DNA synthesis and IL-6 production were also studied using short-term established RCC cell lines. A total of 8 cell lines, 4 RCC cell lines (RC-2, RGB, 14T, 4T) and 4 cell lines established at our laboratory (OCUU-1, 2, 4, 5), as well as 10 short-term established RCC cell lines were used. When TNFalpha and TNFalpha receptor mRNA expression was examined by RT-PCR, p55 TNF receptor mRNA expression was observed while TNFalpha and p75 TNF receptor mRNA expression was not observed in all cell lines. When the effects of TNFalpha on DNA synthesis were studied by [3H]-thymidine incorporation assay, DNA synthesis was induced in RGB, 14T, 4T, OCUU-2 and OCUU-4 by adding 10 and 100 pg/ml of TNFalpha while it was not induced in RC-2 and OCUU-5 at all concentrations. When its effects on cell proliferation were evaluated by MTT assay, cell proliferation was observed in RGB, 14T and 4T at TNFalpha concentration of 10 pg/ml and in RGB, 14T, 4T, OCUU-4 and OCUU-5 at TNFalpha concentration of 100 pg/ml. However, cell proliferation was not detected in RC-2 and OCCU-2 at any concentration. When the effects of TNFalpha on IL-6 production were studied by ELISA, IL-6 production was induced in RC-2, RGB, 14T, OCUU-1, OCUU-2 and OCUU-5 while not in 4T and OCUU-4. When its effects on IL-6 expression were examined by Northern blot analysis, the results were similar to those obtained by ELISA. As for the 10 short-term established RCC cell lines, DNA synthesis and IL-6 production were induced with the addition of TNFalpha. These results suggested that TNFalpha induced cell proliferation in RCC.

Clinicopathological study of renal cell carcinoma.

We investigated the clinicopathological features of 153 patients with renal cell carcinoma who underwent nephrectomy at our institute from April 1992 to August 2001 and evaluated the prognostic significance of various clinical and pathological factors. The prognostic factors were statistically analyzed by the log-rank and generalized Wilcoxon tests, and for the factors that were statistically significant by both analyses, multivariate analysis was conducted by stepwise regression in the Cox proportional hazard model. As a result, the overall 1-year, 3-year, 5-year and 10-year survival rates were 93.5%, 84.3%, 82.9% and 79.1%, respectively. In addition, multivariate analysis showed that regional lymph node metastasis, stage and histological type were independent prognostic factors for renal cell carcinoma. The number of renal cell carcinoma cases incidentally discovered during routine medical check-ups has increased in recent years, and in our study, such incidental tumors accounted for 65.3% of all cases. The 5-year survival rate of incidental tumors was 87.4% as compared to that of non-incidental tumors at 70.1%, and prognosis of incidental tumors was significantly more favorable. However, even in such cases, a favorable prognosis is not necessarily guaranteed according to the presence of regional lymph node metastasis, stage and histological type. Therefore, we conclude that even in incidental tumors, if the patient has unfavorable prognostic factors, post-operative adjuvant therapy should be considered.

Xanthogranulomatous pyelonephritis with acquired cystic disease of the kidney in a haemodialysis patient.

A 54-year-old-female patient who had received regular haemodialysis therapy for 12 years was referred to our hospital for evaluation of a left renal mass. Imaging examinations revealed acquired cystic disease of the kidney (ACDK) and a tumour-like lesion in the left kidney. Because of the preoperative diagnosis of the left renal cell carcinoma, the patient underwent a left nephrectomy. Pathological examination revealed xanthogranulomatous pyelonephritis. It was difficult to distinguish xanthogranulomatous pyelonephritis from renal cell carcinoma in our case, because it is very rare for xanthogranulomatous pyelonephritis to occur in ACDK.

Comparative antitumor activity of 5-fluorouracil and 5'-deoxy-5-fluorouridine in combination with interferon gamma in renal cell carcinoma cell lines.

The treatment of advanced renal cell carcinoma (RCC) remains unsatisfactory, and conventional therapy using interferon (IFN) has shown limited antitumor action. In this study, we examined the combined effects of IFN gamma with 5-fluorouracil (5-FU) as well as 5'-deoxy-5-fluorouridine (5'-DFUR), a prodrug of 5-FU, in vitro. The sensitivity of 4 RCC cell lines to IFN gamma and 5-FU or 5'-DFUR was evaluated using the MTT assay. The IC50 value for the cytostatics was expressed as the concentration at which growth was inhibited by 50% as compared with the control value. Thymidine phosphorylase (TP), the enzyme that converts 5'-DFUR into 5-FU and 5-FU into 5-fluoro-2'-deoxyuridine-5'-monophosphate (FdUMP), was estimated by ELISA. As a result, a significant correlation between TP expression and sensitivity to 5-FU and 5'-DFUR was found in all 4 RCC cell lines. By adding 12.5 U/ml of IFN gamma, which is a concentration that does not affect cell proliferation, the TP expression significantly increased by 1.5 to 3.4 times in the 4 RCC cell lines. Susceptibility to 5-FU and 5'-DFUR was also significantly induced by 1.42 to 2.36 times and 2.9 to 5.71 times, respectively, in the 4 cell lines. These results suggested that TP may be useful as a predictive factor in combination therapy with IFN gamma and 5-FU or 5'-DFUF, which may be a promising treatment for advanced RCC.