Longkun Li

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.

Clinical evaluation of double-pigtail stent in patients with upper urinary tract diseases: report of 2685 cases.

PURPOSE To review the indications, procedures, complications, and related treatments of double pigtail stent (DPS) placement as an adjunct for some types of endoscopic and open urologic surgery. PATIENTS AND METHODS From July 1998 to December 2006, 2413 patients aged 8 to 81 years underwent stent placement (2685 total placements). The indications consisted of ureteroscopic lithotripsy (1984 cases), percutaneous nephrolithotomy (329 cases), ureteral incision to remove calculi (71 cases), extracorporeal shockwave lithotripsy for upper urinary tract calculi (145 cases), ureteropelvic junction obstruction (31 cases), ureterocystoneostomy (29 cases), benign ureteral stenosis (52 cases), extrinsic ureteral stenosis (16 cases), and iatrogenic ureteral trauma (28 cases). DPSs were inserted into the ureter by cystoscopy (115 stents), ureteroscopy (2052 stents), percutaneous nephrostomy (393 stents), or open surgery (125 stents), and were kept inside the body for 28 +/- 1.7 days (range 1-193 days). The mean follow-up period was 31 +/- 1.9 days (range 1-123 days). RESULTS Three hundred sixty-five patients (19.6%) experienced one or more problems during the stenting procedure. The main complications were gross hematuria (385 cases), pain (101 cases), bladder irritation (105 cases), high fever (6 cases), encrustation (53 cases), stent migration (42 cases), and stenosis or restenosis (51 cases). Most of the complications were mild and tolerable, and all were immediately treated appropriately. However, 60 stents had to be removed: 29 for gross hematuria, 18 for pain, 7 for bladder irritation, and 6 for high fever. CONCLUSIONS DPS is a safe and useful adjunct for both endoscopic and open procedures to treat upper urinary tract diseases. Most of the complications of DPS placement can be well managed.

Altered expression of calcium-activated K and Cl channels in detrusor overactivity of rats with partial bladder outlet obstruction

To evaluate the activity of large‐ and small‐conductance calcium‐activated potassium channels (BKCa, SKCa) and calcium‐activated chloride channels (ClCa) in detrusor overactivity (DO) cells after partial bladder outlet obstruction (PBOO) in rats.

Effects of mechanical stretch on interstitial cells of Cajal in guinea pig bladder.

OBJECTIVES C-kit positive interstitial cells of Cajal (ICCs) play an important role in the regulation of the smooth muscle motility. In this study, we investigated the mechanical sensitivity of ICCs in guinea pig bladder and their possible relationship with detrusor overactivity (DO). METHODS Stretch load was performed in vivo in DO models produced by 4 wk of partial bladder outlet obstruction (PBOO). Number and morphology of ICCs were examined using a specific ICC marker, c-kit, immunochemistry staining. The spontaneous and stretch-induced calcium transients (SICT) of ICCs were investigated in cells cultured onto flexible silicone membranes preloaded with the Ca(2+) indicator fluo-4AM in vitro. RESULTS C-kit positive ICCs were mainly located along and between bladder smooth muscle bundles. ICCs in DO bladders displayed more lateral branching with mutual connections. The number of c-kit positive bladder ICCs was increased in the DO group compared with the control group (n = 50, P < 0.05). Cultured ICCs from DO bladders showed spontaneous calcium waves with higher frequency and lower amplitude than those from control bladders (n = 15, P < 0.05). Significant SICT were detected in cultured bladder ICCs. SICT generated in ICCs from DO were more likely to transfer to adjacent smooth muscle cells through cell membrane connection than ICCs from control bladders. CONCLUSIONS Long-term overload tension following PBOO caused changes in morphology, quantity and spontaneous calcium transients of ICCs in guinea pig bladder. Mechanical sensitivity and interaction with SMC of ICCs may contribute to the mechanosensitive conductances in bladder regulation, and may play a role in the pathogenesis of DO.

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.

Zinc levels in seminal plasma and their correlation with male infertility: A systematic review and meta-analysis.

Zinc is an essential trace mineral for the normal functioning of the male reproductive system. Current studies have investigated the relationship between seminal plasma zinc and male infertility but have shown inconsistent results. Hence, we systematically searched PubMed, EMBASE, Science Direct/Elsevier, CNKI and the Cochrane Library for studies that examined the relationship between seminal plasma zinc and male infertility, as well as the effects of zinc supplementation on sperm parameters. Twenty studies were identified, including 2,600 cases and 867 controls. Our meta-analysis results indicated that the seminal plasma zinc concentrations from infertile males were significantly lower than those from normal controls (SMD (standard mean differences) [95% CI] −0.64 [−1.01, −0.28]). Zinc supplementation was found to significantly increase the semen volume, sperm motility and the percentage of normal sperm morphology (SMD [95% CI]: −0.99 [−1.60, −0.38], −1.82 [−2.63, −1.01], and −0.75 [−1.37, −0.14], respectively). The present study showed that the zinc level in the seminal plasma of infertile males was significantly lower than that of normal males. Zinc supplementation could significantly increase the sperm quality of infertile males. However, further studies are needed to better elucidate the correlation between seminal plasma zinc and male infertility.

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.

Roles of stem cell factor on loss of interstitial cells of Cajal in bladder of diabetic rats.

OBJECTIVE To explore the roles of stem cell factor (SCF) on the loss of interstitial cells (ICCs) in the bladder of diabetic rats, which have not been investigated. METHODS The rats were assigned to 3 groups: normal control rats, diabetic rats, and SCF-treated diabetic rats. The diabetic rat model was created using a streptozotocin (60 mg/kg) intraperitoneal injection. The SCF and c-kit levels in bladder tissue were determined using reverse transcriptase-polymerase chain reaction and Western blot analysis. The quantity of ICCs as represented by c-kit-positive cells was examined by image analysis of immunofluorescence staining. RESULTS Compared with the control rats, the diabetic rats exhibited a significant decrease in the SCF levels and c-kit expression and number of ICCs in the bladder tissues. All these impaired parameters were effectively restored to the control level after exogenous SCF treatment. CONCLUSION These findings suggest that the loss of ICCs in the bladder tissue of diabetic rats can be attributed to a deficiency in endogenous SCF. The beneficial effect of exogenous SCF on diabetic depletion of ICCs could provide a theoretical rationale for the use of SCF as a potential therapeutic drug in treating patients with diabetes-related voiding dysfunction.

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.

Identification of T-type Calcium Channels in the Interstitial Cells of Cajal in Rat Bladder

OBJECTIVE To investigate the expression and function of T-type calcium channels in the interstitial cells of Cajal in rat bladders. METHODS Bladders were harvested from Sprague-Dawley rats. The expression of T-type calcium channels subtypes (α1G, α1H, and α1I) in interstitial cells of Cajal were identified by double-labeled immunofluorescence analysis and reverse transcription-polymerase chain reaction analysis in whole mount preparations of rat bladders. The function of T-type calcium channels in freshly isolated interstitial cells of Cajal was assessed by detecting the changes of intracellular calcium ([Ca(2+)](i)) with preloading fluo-3 AM, and by evaluating the changes of the phasic contractions of rat bladder strips after treating with mibefradil and glivec. RESULTS Three T-type calcium channels subtypes, α1G, α1H, and α1I, colocalized with c-kit in bladder interstitial cells of Cajal by double-labeled immunofluorescence analysis, and this was confirmed using reverse transcription-polymerase chain reaction. The T-type calcium channels selective blocker, mibefradil (1 μM), significantly decreased the intracellular calcium concentration ([Ca(2+)](i)) in isolated interstitial cells of Cajal (P < .01) and inhibited the spontaneous phasic contraction of bladder strips (P < .01). Moreover, the c-kit receptor blocker, glivec, significantly decreased the [Ca(2+)](i) of interstitial cells of Cajal further (P < .01) and the spontaneous phasic contraction of bladder strips. CONCLUSION T-type calcium channel subtypes were confirmed to colocalize in interstitial cells of Cajal in rats bladders, which might participate in the spontaneous activity of interstitial cells of Cajal and phasic contractions of bladder strips by modulating [Ca(2+)](i) in interstitial cells of Cajal.