Zhansong Zhou

Detection of Nanobacteria Infection in Type III Prostatitis

OBJECTIVES To investigate the relationship between nanobacterial infection and type III prostatitis. The etiology of type III prostatitis remains unclear to date, although the recently discovered nanobacteria (NB) have been implicated in this disease. METHODS A total of 48 patients with chronic pelvic pain syndrome for whom conventional therapy had failed were selected and randomly divided into two groups, one receiving anti-NB treatment and the other receiving a placebo. The NB were isolated and cultured from expressed prostatic secretions and urine samples before and after treatment. The morphologic features were recorded and 16s rRNA gene expression was determined. The curative effect was evaluated by the NB-positive rate and symptomatic changes using the National Institutes of Health Chronic Prostatitis Symptom Index. RESULTS After anti-NB treatment, the NB-positive rates had decreased from 62.5% to 16.7% in the expressed prostatic secretions and from 12.5% to 0% in the urine samples after prostatic massage (P <0.001). In the patients receiving a placebo, the positive rates had no obvious change in either the expressed prostatic secretions or the urine samples after prostatic massage (P >0.05). The NB were coccoid or coccobacillary and clustered in a diameter of 100 to 500 nm. The BLAST result revealed that the 16s rRNA gene sequence from the NB in the patients with chronic pelvic pain syndrome was 97%, similar to that of the known NB with identity (97%). After anti-NB treatment, the Chronic Prostatitis Symptom Index scores decreased significantly. In contrast, no change in the Chronic Prostatitis Symptom Index scores was seen after placebo treatment. CONCLUSIONS The results of our study have shown that nanobacterial infection might be an important etiologic factor of type III prostatitis. Anti-NB treatment could be an effective therapy against refractory type III prostatitis.

Suppression of CX43 expression by miR-20a in the progression of human prostate cancer.

The aberrant expression of microRNAs (miRNAs) has been found in various types of cancer. The present study found miR-20a to be significantly upregulated in prostate cancer compared with normal prostate tissues. The proliferation and colony formation assays revealed that the downregulation of miR-20a by miR-20a inhibitor suppresses the proliferation of MDA-PCa-2b cells in vitro and also inhibits tumor growth in vivo. Furthermore, a gap junction protein, α 1 (CX43), was identified as a direct target gene of miR-20a. The upregulation of CX43 was detected in MDA-PCa-2b cells after treatment with miR-20a inhibitor both in vitro and in vivo. In conclusion, the findings show that miR-20a significantly contributes to the progression of prostate cancer by targeting CX43.

Celastrol suppresses tumor cell growth through targeting an AR-ERG-NF-κB pathway in TMPRSS2/ERG fusion gene expressing prostate cancer.

The TMPRSS2/ERG (T/E) fusion gene is present in the majority of all prostate cancers (PCa). We have shown previously that NF-kB signaling is highly activated in these T/E fusion expressing cells via phosphorylation of NF-kB p65 Ser536 (p536). We therefore hypothesize that targeting NF-kB signaling may be an efficacious approach for the subgroup of PCas that carry T/E fusions. Celastrol is a well known NF-kB inhibitor, and thus may inhibit T/E fusion expressing PCa cell growth. We therefore evaluated Celastrol’s effects in vitro and in vivo in VCaP cells, which express the T/E fusion gene. VCaP cells were treated with different concentrations of Celastrol and growth inhibition and target expression were evaluated. To test its ability to inhibit growth in vivo, 0.5 mg/kg Celastrol was used to treat mice bearing subcutaneous VCaP xenograft tumors. Our results show Celastrol can significantly inhibit the growth of T/E fusion expressing PCa cells both in vitro and in vivo through targeting three critical signaling pathways: AR, ERG and NF-kB in these cells. When mice received 0.5 mg/kg Celastrol for 4 times/week, significant growth inhibition was seen with no obvious toxicity or significant weight loss. Therefore, Celastrol is a promising candidate drug for T/E fusion expressing PCa. Our findings provide a novel strategy for the targeted therapy which may benefit the more than half of PCa patients who have T/E fusion expressing PCas.

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.

Decreased nanobacteria levels and symptoms of nanobacteria-associated interstitial cystitis/painful bladder syndrome after tetracycline treatment

Introduction and hypothesisThis study was designed to detect whether nanobacteria (NB) reside in urine and bladder tissue samples of patients with interstitial cystitis/painful bladder syndrome (IC/PBS) and whether antibiotic therapy targeting these organisms is effective in reducing NB levels and IC/PBS symptoms.MethodsTwenty-seven IC/PBS patients underwent cystoscopy. Bladder biopsies and urine samples were obtained and cultured for NB, which were identified by indirect immunofluorescent staining and transmission electron microscopy.ResultsEleven bladder samples showed growth of microbes that were identified to be similar to NB. Homologous study of the 16S ribosomal RNA gene suggested that the NB could be the pathogen. For enrolled 11 patients, NB levels decreased dramatically after tetracycline treatment, and they reported significant reduction in the severity of IC/PBS symptoms.ConclusionsA high prevalence of NB was observed in female IC/PBS, and anti-NB treatment effectively improved the symptoms, which suggest that NB may cause some cases of IC/PBS.

Better Compliance Contributes to Better Nocturnal Continence With Orthotopic Ileal Neobladder Than Ileocolonic Neobladder After Radical Cystectomy for Bladder Cancer

OBJECTIVES To investigate, in a randomized controlled study, the degree of continence after the creation of orthotopic ileocolonic and ileal neobladders after cystectomy and to explore a possible mechanism for the difference in continence between these 2 types of orthotopic neobladder. METHODS From 2003 to 2007, 71 male patients underwent orthotopic lower urinary tract reconstruction with either an ileocolonic or ileal neobladder after radical cystectomy. The degrees of continence and voiding patterns were individually evaluated using urodynamic examinations and a detailed patient questionnaire. The abnormal upper tract was evaluated using intravenous urography and ultrasonography. RESULTS Complete daytime continence was achieved in 90.9% and 89.4% of the patients and functional nocturnal continence 48.5% and 76.3% of patients in the ileocolonic neobladder and ileal neobladder groups, respectively. The urodynamic data showed that the initial volume of both the ileocolonic and the ileal neobladder appeared to not be significantly different statistically, although the compliance of the ileocolonic neobladder was lower than that of the ileal neobladder (P < .05). No difference was found in the parameters such as flow rate, urethral profile length, maximal urethral pressure, or neobladder neck pressure between the 2 neobladder types. CONCLUSIONS Although the ileocolonic and ileal neobladders can both achieve a large initial volume, the ileal neobladder has an advantage in the aspect of obtaining satisfactory nocturnal continence because of its greater compliance compared with that of the ileocolonic neobladder.

LXR agonist regulates the carcinogenesis of PCa via the SOCS3 pathway.

Background: Down-regulation of suppressor of cytokine signaling 3 (SOCS3) inhibits prostate cancer (PCa) cell growth. Liver X receptors (LXRs) agonists have been recently introduced for PCa treatment. We postulated that LXR may inhibit the carcinogenesis of PCa via the SOCS3 pathway. Methods: LNCaP cells were cultured and transfected with SOCS3 small-interfering RNA (SOCS3-siRNA) and control small-interfering RNA (control-siRNA). Then cells were treated with LXR activator (GW3965). The expressions of PCa related transcript factors, e.g. transcription 3 (STAT3), nuclear factor kappa B (NF-κB) and activation protein 1(AP1) were detected by western blot assay. In vitro cell proliferation, cell migration, cell invasion and apoptosis were analysed. Nude mice were used for in vivo tumorgenesis. Results: In cells treated with control-siRNA, GW3965 enhanced SOCS3 expression and significantly inhibited the phosphorylation of STAT3, NF-κB and AP1 expressions, accompanied by dramatically reduced cellular proliferation rate, immigration and invasion of cultured cells. In cells treated with SOCS3-siRNA, the inhibitory effects of LXR activator on the phosphorylation of STAT3 and expressions of NF-κB and AP1 were totally abolished. The cell proliferation rate, immigration and invasion were markedly elevated by SOCS3 gene mutation, even with GW3965 treatment. The in vivo tumorgenesis assay showed that GW3965 significantly reduced the tumor volumes in tumor-bearing nude mice receiving saline injection, but failed to limit the tumor volume in tumor-bearing nude mice receiving SOCS3 antibody injection. Conclusion: Our results provide evidence in support of the notion that LXR agonist may regulate the carcinogenesis of PCa via the SOCS3 pathway.

Identification of a Hyperpolarization-activated Cyclic Nucleotide-gated Channel and Its Subtypes in the Urinary Bladder of the Rat

OBJECTIVE To investigate the distribution and effects of hyperpolarization-activated cyclic nucleotide-gated (HCN) channel and its isoforms in bladder, especially in bladder interstitial cells of Cajal (ICC). METHODS Four HCN isoforms were detected in bladder tissue from rats using reverse transcription-polymerase chain reaction and Western blotting. The HCN1 subtype was observed in bladder ICCs by double-labeled fluorescence. The effect of the HCN blocker, ZD7288, was investigated using the bladder smooth muscle strip test. RESULTS HCN1-4 isoforms were all identified in bladder ICCs using reverse transcription-polymerase chain reaction and Western blotting. Based on our semiquantitative analysis, HCN1 was found to be the most prominent isoform. The expression of HCN1 was confirmed in bladder ICCs by double-labeled fluorescence through colabeling of HCN1 and kit (CD117). ZD7288 significantly decreased the bladder excitation. CONCLUSION All 4 HCN channel isoforms exist in the bladder, and they affect the bladder excitation, presumably via bladder ICCs.

Nanobacteria: A Possible Etiology for Type III Prostatitis

PURPOSE Nanobacteria are thought to be a pathopoiesis bacterium in urological disease. We observed pathological changes in nanobacteria infected prostates in Sprague-Dawley(R) rats and investigated the possible etiological relationships of nanobacteria and type III prostatitis. MATERIALS AND METHODS We randomized 40 adult male Sprague-Dawley rats each to the control and model groups. Rat prostate infection models were reproduced by infusing nanobacteria suspension transurethrally. Rats were sacrificed 1, 2, 4 and 8 weeks later, respectively. Prostatic pathology, and the cytokines interleukin-1beta and tumor necrosis factor-alpha were assessed. Nanobacteria isolation, culture and characterization were also analyzed. RESULTS In model rats we observed prostatic acute inflammatory changes 1 to 2 weeks after nanobacteria infusion and chronic inflammatory changes after 4 weeks. At 8 weeks we noted microcalculous formation in the prostatic glandular cavity in 7 of the 10 model rats, which was not seen in controls. Interleukin-1beta and tumor necrosis factor-alpha in prostatic tissues were higher in model rats than in controls at different time points (p <0.01). In model rats interleukin-1beta and tumor necrosis factor-alpha were higher 2 weeks after infusion than at 1, 4 and 8 weeks (p <0.05). Prostatic tissue was nanobacteria positive in 35 model rats and in 0 controls. CONCLUSIONS Nanobacteria may be an important etiological factor for type III prostatitis.

Transurethral Endoscopy Technique with a Ureteroscope for Diagnosis and Management of Seminal Tracts Disorders: A New Approach

PURPOSE To apply a transurethral endoscopic technique for examining and managing suspicious distal seminal tracts disorders with a ureteroscope. PATIENTS AND METHODS Sixteen patients with distal seminal tracts disorders underwent transurethral endoscopy through the distal seminal tracts using a semirigid ureteroscope. Of the 16 patients, 6 had suspected hemospermia, 4 spermatocele, and 6 male infertility. RESULTS The ejaculatory duct, seminal vesicle, and ampulla of the vas deferens were observed under direct vision with the ureteroscope. The vas deferens was investigated by cannulation with a guidewire or an epidural anesthesia catheter. Four patients received a diagnosis of spermatocele, four seminal vesiculitis, and three vas deferens obstruction. All patients received appropriate treatment. The remaining five patients had no anatomic disorders. All patients received careful postoperative observation and treatment, and were monitored for at least 3 months. Three patients had postoperative discomfort in the perineal region. There were no further complications. CONCLUSIONS This new technique with the ureteroscope enables diagnosis and management of distal seminal tracts disorders through the normal anatomic tract. This endoscopic technique can be performed easily with minimal complications.