[Function of bladder smooth muscle autophagy in cyclophosphamide-induced cystitis].

OBJECTIVE: To explore the existence of bladder smooth muscle autophagy and its function in cyclophosphamide (CYP) -induced cystitis. METHODS: A total of 30 Sprague-Dawley rats were randomized into control (n = 6) and experimental groups (4 h, 48 h, 72 h, 96 h, n = 24) . The experimental group received a single intraperitoneal injection of 150 mg/kg CYP in 0.6 ml saline while the control group had the same volume of saline injection. The changes of microtubule-associated protein light chain 3 (LC3), interleukin-6 (IL-6), IL-1ß and tumor necrosis factor-alpha (TNF-α) proteins in bladder muscular layer were measured with Western blot. Co-location of LC3 and alpha-small muscle action (α-SMA) was detected with double-labeled immunofluorescence. Autophagic vacuoles were observed with electron microscopy. The changes in bladder inflammation were measured by hematoxylin and eosin staining. RESULTS: Autophagy was confirmed in detrusor myocytes with electron microscope, LC3 Western blot and double-labeled immunofluorescence. As compared with the control group, the expression level of LC3 in experimental groups decreased (0.08 ± 0.01 vs 0.13 ± 0.01, P < 0.05) at 4 h and increased 1.19, 0.75 and 0.20 fold (0.29 ± 0.04 vs 0.13 ± 0.01, P < 0.05; 0.23 ± 0.03 vs 0.13 ± 0.01, P < 0.05;0.16 ± 0.03 vs 0.13 ± 0.01, P > 0.05) at 48, 72 and 96 h respectively.Electron microscopy and double-labeled immunofluorescent also supported the above change of LC3 by Western blot.In addition, as compared with the control groups, IL-6, IL-1ß and TNF-α and bladder histological scores of bladder muscular layer significantly increased in experimental groups (all P < 0.05) . CONCLUSIONS: Autophagy exists in bladder smooth muscle. In cyclophosphamide-induced cystitis, bladder smooth muscle autophagy activation may not be sufficient to protect bladder muscle layer inflammation in rats.

[Perineal urethrostomy plus secondary urethroplasty for ultralong urethral stricture: clinical outcomes and influence on the patient's quality of life].

OBJECTIVE: To investigate the outcomes of perineal urethrostomy plus secondary urethroplasty for ultralong urethral stricture and assess its influence on the patient's quality of life. METHODS: We retrospectively analyzed 54 cases of ultralong urethral stricture treated by perineal urethrostomy from 2000 to 2010. The mean age of the patients was 40 years, and the average length of stricture was 6.5 cm. We evaluated the patients'quality of life by questionnaire investigation and the clinical outcomes based on IPSS, Qmax, the necessity of urethral dilation and satisfaction of the patients. RESULTS: The mean Qmax of the 54 patients was (14.0 +/- 4.7) ml/min. Of the 34 cases that underwent secondary urethroplasty, 22 (64.7%) achieved a mean Qmax of (12.0 +/- 3.5) ml/min, 8 (23.5%) needed regular urethral dilatation and 4 (11.8%) received internal urethrotomy because of restenosis. IPSS scores were 5.4 +/- 2.1 and 8.5 +/- 5.8 after perineal urethrostomy and secondary urethroplasty, respectively. Fifty of the total number of patients (92.6%) were satisfied with the results of perineal urethrostomy, and 22 of the 34 (64.7%) with the results of secondary urethroplasty. CONCLUSION: Perineal urethrostomy plus secondary urethroplasty is safe and effective for ultralong urethral stricture, and affects very little the patient's quality of life.

Prognostic value of vascular endothelial growth factor expression in patients with prostate cancer: a systematic review with meta-analysis.

BACKGROUND: The vascular endothelial growth factor (VEGF) mediates vasculogenesis and angiogenesis through promoting endothelial cell growth, migration and mitosis, and has involvement in cancer pathogenesis, progression and metastasis. However, the prognostic value of VEGF in patients with prostate cancer remains controversial. OBJECTIVES: The aim of our study was to evaluate the prognostic value of VEGF in prostate cancer, and summarise the results of related research on VEGF. METHODS: In accordance with an established search strategy, 11 studies with 1,529 patients were included in our meta-analysis. The correlation of VEGF-expression with overall survival and progression-free survival was evaluated by hazard ratio, either given or calculated. RESULTS: The studies were categorized by introduction of the author, demographic data in each study, prostate cancer-relatived information, VEGF cut-off value, VEGF subtype, methods of hazard ratio (HR) estimation and its 95% confidence interval (CI). High VEGF-expression in prostate cancer is a poor prognostic factor with statistical significance for OS (HR=2.32, 95%CI: 1.40-3.24). However, high VEGF-expression showed no effect on poor PFS (HR=1.30, 95%CI: 0.88-1.72). Using Begg's, Egger's test and funnel plots, we confirmed lack of publication bias in our analysis. CONCLUSION: VEGF might be regarded as a prognostic maker for prostate cancer, as supported by our meta-analysis. To achieve a more definitive conclusion enabling the clinical use of VEGF in prostate cancer, we need more high-quality interventional original studies following agreed research approaches or standards.

Nanobacteria may be linked to testicular microlithiasis in infertility.

Testicular microlithiasis (TM) in infertility is an uncommon pathologic condition of unclear etiology that is characterized by calcium deposits within the seminiferous tubules. Nanobacteria (NB), as novel microorganisms mediating tissue calcification, have been discovered in some diseases. In this study, we hypothesized that NB may participate in the pathogenesis of TM, particularly in infertility. Seventeen infertility patients with TM detected by scrotal color Doppler ultrasonography and 17 infertility patients without TM as controls were enrolled in the study. The NB were isolated and cultured from semen samples and urine samples. After 3 to 6 weeks of culture, 10 of 17 (58.8%) semen samples and 2 urine samples from infertile patients with TM showed the growth of white granular microbes that firmly attached to the bottom of the culture flask and were visible to the naked eye. In the control group, only 1 of 17 (5.9%) semen samples from infertile patients without TM showed the growth of white granular microbes. The cultured microbes were identified by indirect immunofluorescent staining (IIFS), transmission electron microscopy (TEM), and 16s rRNA gene expression. IIFS and TEM revealed NB to be coccoid and 100 to 500 nm in diameter. The BLAST result revealed that the 16s rRNA gene sequence from the cultured microbes was 97% the same as that of the known NB. Our results showed that NB may be linked to the development of TM, which may provide a potential target for the diagnosis and treatment of infertility with TM.

[Correlation between activation of L5-S2 spinal cord astrocytes and effect of substance P in chronic prostatitis pain].

OBJECTIVE: To observe the expressions of the substance P (SP) mRNA and neurokinin-1 receptor (NK-1R) in the posterior horn of the L5 - S2 spinal cord in the rat model of chronic prostatitis pain, and to investigate the changes in the activation of astrocytes and influence of SP on this activation in rat spinal cord astrocytes cultured in vitro. METHODS: The rat model of chronic prostatitis pain was established by injection of complete Freund's adjuvant (CFA) and assessed by the tail flick threshold test, the control rats injected with sodium chloride and all observed at 0, 14 and 28 days. Changes in the expressions of SP mRNA, NK-1R, glial fibrillary acidic protein (GFAP), tumor necrosis factor-alpha (TNF-alpha) and inducible nitric oxide synthase (iNOS) in the posterior horn of the L5 - S2 spinal cord were detected by RT-PCR and Western blot. Rat spinal cord astrocytes were cultured in vitro and divided into a control group, cultured with ITS cell culture fluid, and two experiment groups, with Group 1 stimulated with SP at the concentration of 10(-9) - 10(-6) mol/L for 12 hours followed by determination of the expressions of TNF-alpha, IL-1beta, NO and NOS by ELISA and nitrate reductase and colorimetric methods, and Group 2 at 10(-7) mol/L for 0, 24, 48 and 72 hours followed by detection of the GFAP expression by Western blot. RESULTS: The expressions of SP mRNA, NK-1 R, GFAP, TNF-alpha and iNOS in the posterior horn of the L5 - S2 spinal cord were obviously higher in the rat prostatitis pain models than in the controls, successively higher at 28 than at 14 and 0 d (P < 0.01), and so was the expression of GFAP at 28 than at 14 d in the experiment groups (P < 0.05). SP induced a gradual increase at 10(-7) mol/L in the expression of GFAP in the spinal cord astrocytes at 0 -72 h, significantly different from that of the control group (P < 0.01), and it promoted the excretion of TNF-alpha and IL-1beta and the activity of NO and NOS at 10(-9) - 10(-6) mol/L at 12 h in a concentration-dependent manner, with marked differences between the experiment and control groups (P < 0.01, P < 0.05). But a decreased excretion of IL-1 beta was observed in the 10(-6) mol/L group, though with no significant difference from the control (P > 0.05). CONCLUSION: Chronic prostatitis pain could upregulate the expressions of the excitatory transmitter SP and receptor in the L5 - S2 spinal cord, and result in the activation of astrocytes and increased excretion of proinflammatory cytokines, which may be associated with the persistence and generalization of prostatitis pain.

[Morphological study on the role of ICC-like cells in detrusor neuro-modulation of rat urinary bladder].

OBJECTIVE: To investigate the role of ICC-like cells in bladder neuromodulation in rat urinary bladder. METHODS: 14 SD rats and 1 guinea pig were sacrificed in this study. The ultra structural relationships among interstitial cells, nerves and detrusor smooth muscle cells (DSMCs) of urinary bladder were investigated by transmission electron microscopy (TEM). c-kit immunofluorescence was used to identify ICC-like cells in SD rat urinary bladder and the structural relationship between ICC-like cells and nerve terminals was studied by immunofluorescence (double-label). RESULTS: Gap junction between ICC-like cells and DSMCs was confirmed by TEM. ICC-like cells were very close apposition with nerve terminals under TEM. ICC-like cells were identified to exist in sub-urothelium layer, along the longitude of smooth muscle bundles and among detrusor smooth muscle in SD rat urinary bladder by c-kit immunofluorescence. Double-labeled tissue with c-kit and PGP9.5 antibodies also showed that ICC-like cells were very close apposition with nerve terminals in SD rat bladder. CONCLUSIONS: Morphological study indicated that ICC-like cells in rat urinary bladder may play an important role in detrusor neuromodulation. Further study on function will be helpful for elucidating the mechanism of bladder neuromodulation clearly.