Quantitative assessment of bladder neck compliance by using transvaginal real-time elastography of women.

To assess the feasibility of using ultrasound real-time elastography (RTE) to measure bladder neck compliance, we performed real-time elastography measurements by manually applying repetitive compression with the transducer on the scan position of the bladder neck. Instant elastography index (EI) and mean EI of anterior and posterior lips of the bladder neck were calculated. The EI values of anterior and posterior lips of the bladder neck were analyzed in relation to age, body surface area, body mass index, detrusor wall thickness and length, width and thickness of the bladder neck in healthy women. The intra-observer and inter-observer repeatability of measurements in different parts of the bladder neck were assessed using intra-class correlation coefficients with 95% confidence intervals and Bland-Altman analysis. There were no statistically significant differences between elastography measurements made by the same or two different observers in each area measured. There was no significant difference between anterior and posterior lip thickness of the bladder neck. The distribution of the elastography measurements indicated that the anterior lip of the bladder neck was slightly harder than the posterior lip. On the whole, from the results of the study, it was clear that EIs of the bladder neck were related to age in healthy women. Stepwise multiple regression analysis results revealed that age was the only independent factor modulating compliance of the bladder neck in healthy women. It is possible to provide a reproducible semi-quantification of real-time elastography in bladder neck compliance.

TLR4 mediates MAPK-STAT3 axis activation in bladder epithelial cells.

The role of Toll-like receptor 4 (TLR4) in immune cells is well characterized, but its biological properties in bladder epithelial cells (BECs), especially reciprocal crosstalk between mitogen-activated protein (MAP) kinase pathway and signal transducer and activator of transcription (STAT)3-mediated signal transduction elicited by TLR4 have not been demonstrated so far. The present studies were to demonstrate the signal transduction and inflammatory cytokine response elicited through activation of TLR4 in BECs with a special focus on the crosstalk between the MAPK-pathway and STAT3-mediated signals and its regulatory relevance for the inflammatory response towards lipopolysaccharide (LPS). We selected human bladder cancer T24 cell line in the present study and examined its expression of TLR4 by reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemistry. The expression of p38, extracellular signal regulated kinase (ERK), c-Jun NH2-terminal kinase (JNK), and STAT3 were performed by RT-PCR, quantitative PCR, and Western blotting. Signal transduction was analyzed by Western blotting. Interleukin-6 (IL-6) and interleukin-10 (IL-10) secretion in culture supernatants were tested by human enzyme-linked immunosorbent assay (ELISA) kit. BECs of rat infection in vivo model and patients with cystitis glandularis (CG) were analyzed as described above. Our study demonstrated that TLR4 was significantly upregulated following LPS treatment, with the maximum mRNA expression occurring at 4 h after stimulation. Activation of TLR4 signaling by LPS resulted in phosphorylation of MAPK and STAT pathways and upregulation of IL-10 in dose- and time-dependent manners in T24 cells. Pretreatment of cells with SB203580 (inhibitor of p38) and SP600125 (inhibitor of JNK) attenuated LPS-induced IL-10 expression, whereas it markedly inhibited the STAT3 expression. IL-10 mRNA expression was increased in inflamed lesions compared to noninflamed tissue in rats and patients with CG disease. Our results demonstrate that activation of TLR4 signaling in BECs induces IL-10 expression via activation of p38 and JNK, and the activation of STAT-3 was upregulated. Our data indicated that the reciprocal crosstalk between the MAPK pathway and STAT3-mediated signal transduction forms a critical axis successively activated by LPS in BECs.