[Dynamic change in the gene expression profile of rat benign prostate hyperplasia tissue after complete denervation].

OBJECTIVE: To investigate the dynamic change in the gene expression profile of the rat BPH tissue with progressive atrophy after complete denervation. METHODS: Twelve 29-week-old male rats with spontaneous hypertension and spontaneously developed BPH were used for this study, of which 3 were included in the control (C) group and the other 9 underwent complete denervation of the prostate. At 3, 7 and 11 days after operation (the D3, D7 and D11 groups), all the rats were sacrificed and their ventral prostatic lobes harvested for histopathological examination and RNA extraction, and the RNA samples were subjected to whole genome microarray of the expression profile, followed by real-time RT-PCR validation and bioinformatics analysis. RESULTS: Progressive atrophy of the BPH tissue was observed in the rats after complete denervation. Whole genome microarray of the expression profile was successfully performed for all the samples, and its reliability validated by real-time RT-PCR of 6 differentially expressed genes selected randomly. Hierarchical clustering analysis showed 108 up-regulated and 175 down-regulated genes in the differentially expressed ones between the D3 and C groups, 462 up-regulated and 189 down-regulated in those between the D7 and C groups, and 548 up-regulated and 256 down-regulated in those between the D11 and C groups. GO functional enrichment analysis indicated that the genes in each differentially expressed gene set participated in hundreds of molecular functions, biological processes and cellular components, while pathway enrichment analysis showed their involvement in hundreds of signaling pathways, of which many were enriched simultaneously in each differentially expressed gene set and ranked as the most enriched ones, and most of the genes involved were up-regulated and related with the activation of the complement system. CONCLUSIONS: Large numbers of abnormally expressed genes are involved in the progressive atrophy of rat BPH tissue after complete denervation, and these genes participate in hundreds of molecular functions, biological progresses, cellular components and signaling pathways. Abnormal activation of the complement system may play an important role in the progressive atrophy of the BPH tissue.

Correlation between Cholinergic Innervation, Autophagy, and Etiopathology of Benign Prostatic Hyperplasia.

BACKGROUND: Whether cholinergic innervations and/or autophagy have a role in the etiopathology of benign prostatic hyperplasia (BPH) is still unknown. This study aimed to investigate the role of cholinergic innervation and autophagy in the etiopathology of BPH. METHODS: Male, 13-week-old spontaneous hypertension rats (spontaneous BPH animal model) were divided into three groups: an experimental group (EG, n = 24), a control group (CG, n = 24), and a normal control group (NC, n = 10). The EG animals were intragastrically injected with tolterodine (3.5 mg/kg, twice a day), CG animals were intragastrically injected with physiological saline, and the NC animals did not receive any treatment. Rats were sacrificed every 4 weeks, and the prostatic gross morphological changes, wet weight/body weight (ww/bw), dry weight/wet weight (dw/ww), histological changes, ultrastructural changes, and LC3 immunohistochemistry were continuously observed and compared. RESULTS: The gross morphological and ww/bw changes in the three groups were similar at every stage. The dw/ww (mg/mg) values of the EG at week 17, 21, 25, and 29 were 0.1478 ± 0.0034, 0.1653 ± 0.0036, 0.1668 ± 0.0045, and 0.1755 ± 0.0034, respectively, and the CG values were 0.1511 ± 0.0029, 0.1734 ± 0.0020, 0.1837 ± 0.0052, and 0.1968 ± 0.0045, respectively. The difference between EG and CG for dw/ww showed statistical significance after 21 weeks of age (week 21: P= 0.016, week 25: P= 0.008, and week 29: P= 0.001). Both EG and CG, prostatic glandular epithelial cell proliferation, and secretory function improved with age, but in EG, these improvements were slower than those in CG, and all the differences were statistically significant after 21 weeks. An increasing number of autophagosomes in the prostatic glandular cell cytoplasm, attenuation of LC3-I immunohistochemical staining, enhancement of LC3-II staining, and the ratio of LC3-II/LC3-I staining were all progressive in both groups, but the rate of change in EG was faster than that in CG, and these differences gained statistical significance after 25 weeks. Comparisons with regard to the above indexes between CG and NC showed no statistical significance at any stage. CONCLUSIONS: Cholinergic innervations and activation of autophagy appear to have important functions in the etiopathology of BPH. Drug-mediated blockade of cholinergic innervations could delay the physiopathology processes. Moreover, overactivation of autophagy may also play an important role in this delay.

[A dosage regimen of M-receptor blocker after TURP for severe BPH with predominant urine storage symptoms].

OBJECTIVE: To study the dosage regimen of oral M-receptor blocker following transurethral resection of the prostate (TURP) for severe benign prostate hyperplasia (BPH) with predominant urine storage period symptoms (USPSs) and its clinical effect. METHODS: Severe BPH patients with predominant USPSs received oral tolterodine (2 mg q12d or 4 mg qd) 6 hours after TURP for 4 weeks. The medication continued for another 2 weeks in case of recurrence of USPSs or until the 12th week in case of repeated recurrence. Before and at 1, 4, 8 and 12 weeks after TURP, we analyzed the International Prostate Symptoms Score (IPSS), quality of life (QoL) score, maximum urinary flow rate (Qmax), and postvoid residual volume (PVR) of the patients. RESULTS: Complete clinical data were collected from 106 cases, of which 33 achieved successful drug withdrawal with no aggravation of USPSs at 4 weeks after TURP, 51 at 6－8 weeks, 13 at 10－12 weeks, and 9 needed medication after 12 weeks. Before and at 1, 4, 8 and 12 weeks after TURP, the total IPSSs were 25.33 ± 3.45, 19.33 ± 3.62, 11.56 ± 2.45, 8.38 ± 2.0 and 7.74 ± 1.87, those in the urine storage period were 11.97 ± 1.53, 10.76 ± 1.82, 6.16 ± 1.22, 4.08 ± 1.19 and 3.91 ± 1.15, those at urine voiding were 9.80 ± 1.60, 5.59 ± 1.45, 3.40 ± 0.92, 2.85 ± 0.71, and 2.61 ± 0.67, and the QoL scores were 4.70 ± 0.78, 3.92 ± 0.75, 2.55 ± 0.74, 1.83 ± 0.72 and 1.66 ± 0.75, respectively, with statistically significant differences between the baseline and the scores at 1 and 4 weeks (P <0.01) but not at 8 or 12 weeks (P >0.05). Qmax and PVR were improved progressively and significantly at 1 and 4 weeks (P <0.01) but not at 8 or 12 weeks (P >0.05). CONCLUSIONS: Four to eight weeks of oral administration of M-receptor blocker may be an effective dosage regimen for severe BPH with predominant USPSs after TURP.

Efficacy and Safety of Medium-to-long-term Use of Tolterodine Extended Release with or without Tamsulosin in Patients with Benign Prostate Hyperplasia and Larger Prostate Size: A Double-blind, Placebo-controlled, Randomized Clinical Trial.

BACKGROUND: The medium-to-long-term use of antimuscarinics alone or in combination with an α-blocker in men with an enlarged prostate is still controversial. This double-blind, placebo-controlled, randomized clinical trial aimed to investigate the efficacy and safety of medium-to-long-term use of tolterodine extended release (ER) with or without tamsulosin in patients with benign prostate hyperplasia (BPH) and larger prostate size. METHODS: Totally, 152 patients (age ≥50 years) with BPH, International Prostate Symptom Score (IPSS) ≥12, quality-of-life (QoL) score ≥3, and total prostate volume ≥25 ml were enrolled in this study. The patients were randomized into four groups (n = 38 in each) to receive tolterodine ER placebo plus tamsulosin placebo, 0.2 mg tamsulosin plus tolterodine ER placebo, 4 mg tolterodine ER plus tamsulosin placebo, or tolterodine ER plus tamsulosin once daily for 24 weeks. IPSS (total, storage, and voiding subscales), QoL, maximum urinary flow rate (Qmax), and postvoid residual volume (PVR) were collected at baseline, and at weeks 4, 12, and 24. RESULTS: Compared with placebo, tolterodine ER plus tamsulosin significantly improved total IPSS (-7.15, -12.20, and -14.66 vs. -3.51, -5.78, and -7.23), storage IPSS (-3.56, -5.63, and -6.66 vs. -1.52, -1.21, and -2.43), voiding IPSS (-2.88, -5.10, and -6.48 vs. -1.52, -3.03, and -2.97), QoL (-1.21, -2.40, and -3.21 vs. -0.39, -1.41, and -1.60), Qmax (2.21, 7.97, and 9.72 ml/s vs. 2.15, 2.44, and 2.73 ml/s), and PVR (-17.88, -26.97, and -27.89 ml vs. -12.03, -11.16, and -16.73 ml) at weeks 4, 12, and 24, respectively; the differences were all statistically significant (P < 0.05). Adverse events (AEs) were not increased with treatment progression. Tolterodine ER alone did not improve total IPSS (-4.61, -6.79, and -5.70), voiding IPSS (-0.64, -1.83, and -1.45), QoL (-0.69, -1.21, and -1.41), or Qmax(-0.79, 2.83, and 1.11 ml/s), compared with placebo (all P > 0.05). However, a gradual increase in PVR (10.03, 10.41, and 12.89 ml) and more urinary AEs suggestive of urinary retention (11/38 vs. 4/38) were observed. CONCLUSION: Medium-to-long-term use of tolterodine ER plus tamsulosin should be recommended in patients with BPH and an enlarged prostate volume. TRIAL REGISTRATION: www.chictr.org.cn, ChiCTR-TRC-09000596; http://www.chictr.org.cn/showproj.aspx?proj=8939.

Proficiency of virtual reality simulator training in flexible retrograde ureteroscopy renal stone management.

BACKGROUND: Minimally invasive flexible ureteroscopy techniques have widely adopted in the management of patients with renal stones. We performed this study to investigate the value of virtual reality simulator training in retrograde flexible ureteroscopy renal stone treatment for catechumen. METHODS: Thirty catechumen, included 17 attending physicians and 13 associate chief physicians, were selected for study. The trainees first underwent 1-hour basic training to get familiar with the instrument and basic procedures, then followed by 4-hour practice on virtual reality simulators. Before and after the 4-hour training, all trainees undertake an assessment with task 7 program (right low pole calyces stone management). We documented for each trainee the total time of procedure, time of progressing from the orifice to stone, stone translocation and fragmentation time, laser operate proficiency scale, total laser energy, maximal size of residual stone fragments, number of trauma from the scopes and tools, damage to the scope and global rating scale (GRS). The proficiency of this training program was analyzed by the comparison of the first and second assessment outcomes. RESULTS: Significant improvement was observed in retrograde flexible ureteroscopy management of renal stone on virtual reality simulators after finishing the 4 hour special-purpose training. This was demonstrated by improvement in total procedure time ((18.37±2.59) minutes vs. (38.67±1.94) minutes), progressing time from the orifice to stone ((4.00±1.08) minutes vs. (13.80±2.01) minutes), time of stone translocation ((1.80±0.71) minutes vs. (6.57±1.01) minutes), fragmentation time ((4.43±1.25) minutes vs. (13.53±1.46) minutes), laser operate proficiency scale (8.47±0.73 vs. 3.77±0.77), total laser energy ((3231.6±401.4) W vs. (5329.8±448.9) W), maximal size of residual stone fragments ((2.66±0.39) mm vs. (5.77±0.63) mm), number of trauma from the scopes and tools (3.27±1.01 vs. 10.37±3.02), damage to the scope (0 vs. 0.97±0.76) and GRS (29.27±2.95 vs. 9.87±2.21). The differences between the first and the second assessment were all statistically significant (all P < 0.01). CONCLUSION: The virtual reality simulator training program can help the trainees to rapidly improve their retrograde flexible ureteroscopy skill in renal stone treatment.

[Value of laparoscopic virtual reality simulator in laparoscopic suture ability training of catechumen].

OBJECTIVE: To investigate the value of laparoscopic virtual reality simulator in laparoscopic suture ability training of catechumen. METHODS: After finishing the virtual reality training of basic laparoscopic skills, 26 catechumen were divided randomly into 2 groups, one group undertook advanced laparoscopic skill (suture technique) training with laparoscopic virtual reality simulator (virtual group), another used laparoscopic box trainer (box group). Using our homemade simulations, before grouping and after training, every trainee performed nephropyeloureterostomy under laparoscopy, the running time, anastomosis quality and proficiency were recorded and assessed. RESULTS: For virtual group, the running time, anastomosis quality and proficiency scores before grouping were (98 ± 11) minutes, 3.20 ± 0.41, 3.47 ± 0.64, respectively, after training were (53 ± 8) minutes, 6.87 ± 0.74, 6.33 ± 0.82, respectively, all the differences were statistically significant (all P < 0.01). In box group, before grouping were (98 ± 10) minutes, 3.17 ± 0.39, 3.42 ± 0.67, respectively, after training were (52 ± 9) minutes, 6.08 ± 0.90, 6.33 ± 0.78, respectively, all the differences also were statistically significant (all P < 0.01). After training, the running time and proficiency scores of virtual group were similar to box group (all P > 0.05), however, anstomosis quality scores in virtual group were higher than in box group (P = 0.02). CONCLUSION: The laparoscopic virtual reality simulator is better than traditional box trainer in advanced laparoscopic suture ability training of catechumen.

[Differential expression of hyaluronic acid synthase family in human bladder transitional cell carcinoma and its clinical significance].

OBJECTIVE: To investigate the differential expression of the hyaluronic acid synthase (HAS) family in human bladder transitional cell carcinoma (BTCC) and its potential clinical significance. METHODS: The relative quantitative detection of the expression of HAS isoforms (HASs) was performed in 78 human BTCC tissues (mRNA & protein) and 12 normal human bladder mucosa (protein) by real-time RT-PCR and Western blot, and the results were statistically analyzed according to the clinical data. RESULTS: All the BTCC tissues expressed three HAS isoform mRNA and protein, but to a different extent, as for mRNA, HAS3 > HAS2 > HAS1 (P < 0.001), with a significant difference in HAS1/HAS2, HAS1/HAS3 and HAS2/HAS3 (P = 0.003, < 0.001, 0.006, respectively). Among the proteins, the HAS2 expression was the highest, with a significant difference in HAS1/HAS2, and HAS2/HAS3 (P = 0.004, 0.001, respectively), but not in HAS1/HAS3. The elevation of HAS1 mRNA and protein expression was significantly related with the tumor malignancy, tumor initial onset/recurrence, T1/T2 and T1/T3-4 stags, and tumor grading (P = 0.02, < 0.001, 0.038, < 0.001; 0.025, 0.031, 0.023, 0.002; respectively). The HAS2 mRNA expression was significantly related with tumor size (diameter ≤ 3.0 cm/> 3.0 cm), tumor number (single or multiple), tumor initial onset/recurrence, T-staging, and histopathological differentiation (low grade/high grade) (P = 0.012, 0.004, < 0.001, < 0.001, < 0.001, respectively), but its protein expression was not significantly different in all subgroups except with the tumor size (mass diameter > 3.0 cm/≤ 3.0 cm). However, HAS3 mRNA and protein expression had no significant difference among all the subgroups. In normal human bladder mucosa, no HAS expressions were detected. CONCLUSIONS: The abnormally high expression of the HASs further indicate the reliability of hyaluronan as a urinary marker for human BTCC. Compared with HAS1 and HAS3, HAS2 as a marker may have more usefulness in studies on human BTCC carcinogenesis or development. The high expression of HAS1 protein seems to play a more important role in the BTCC tumorigenesis, and may indicate a poor prognosis of the BTCC patients.

Correlation between hyaluronic acid,hyaluronic Acid synthase and human renal clear cell carcinoma.

OBJECTIVE: To study the correlation between hyaluronic acid (HA), hyaluronic acid synthase (HAS) and human renal clear cell carcinoma (RCCC). METHODS: The expression of three HAS isoforms' gene and HA in 93 RCCC tissues, 27 nephridial tissues by the side of RCCC from two hospitals were measured with Real-Time RT-PCR、Western Blot and immunohistochemical methods and analyzed. RESULTS: All RCCC and adjacent normal tissues expressed three HASs' mRNA & protein; at the mRNA level, both RCCC and adjacent normal tissues, expressed more HAS3 than HAS1 or HAS2, their differences were statistically significant (all P values <0.05); but, at the protein level, all HAS isoforms presented the equivalent expression. Compared with the adjacent non-neoplastic kidney tissues, the expression of all HAS isoforms' mRNA in RCCC tissues were increased evidently and their differences were significant (all P values <0.0001); but at the protein level, only the expression of HAS3 increased evidently (P=0.022). In all adjacent normal tissues, more than 80% renal tubular cells strongly expressed HA, however, only the minority RCCC cases (16/93) presented weakly positive HA staining in few cancer nests (5%-30%), the difference were significant (P<0.0001). In RCCC tissues subgrouped according to clinical stage, pathological grade, lymphatic metastasis or not and distant metastasis or not, the HASs' mRNA & protein differential expression all had no statistical significance (all P values >0.05). CONCLUSION: Different from other malignancy, HA and HASs (except for HAS3) may not play important roles in the biological progress of human RCCC.

[Pathological changes of benign hyperplastic prostate after removal of innervation of cholinergic pelvic nerve: experiment with spontaneous hypertension rats].

OBJECTIVE: To study the pathological change of benign hyperplastic prostate after removal of the innervation of cholinergic parasympathetic pelvic nerve. METHODS: Sixty-five male spontaneous hypertension rats (SHRs) were randomly assigned into 3 groups: operation group (n = 30) undergoing truncation of bilateral originating branches of parasympathetic pelvic nerve of major pelvic ganglion (MPG) followed by cystostomy, sham operation group (operation control group, n = 30) undergoing cystostomy, and normal control group (n = 5) not undergoing operation. 3, 7, 11, 15 and > or = 21 days after operation 6 rats from the 2 operation groups and 1 from the control group were sacrificed to observe the gross morphology and histological and cellular changes of the prostate glands. RESULTS: The prostate of the operation group on post-operational day 7 showed mild granular solidification and such change progressed gradually over time, the ratio of prostate wet weight/rat body weight was (0.4764 +/- 0.0125) mg/g on day 3, then gradually decreased, and became (0.2749 +/- 0.0197) mg/g > or = 21 days post-operationally; while the ratio of prostate tissue dry weight/wet weight on day 3 was (0.1966 +/- 0.0062), then gradually increased, and became (0.2596 +/- 0.0035) > or = 21 days post-operationally. HE staining showed that the glandular structure gradually became dilated and rounded, with accumulation of prostatic fluid. The glandular epithelial cells showed gradual degeneration, necrosis, and detachment. The glandular epithelium became progressively thinner, the smooth muscles elongated and thinned progressively, and the stromal components showed mild to moderate overgrowth. Electron microscopy showed that the glandular cells gradually underwent vacuolar degeneration and the structures of the basement membrane became fuzzy. The smooth muscle cells degenerated mildly, and the fibroblasts and collagenous fibers in the stroma overgrew slowly. All these histological changes were not found in the sham operation control and normal control groups. CONCLUSION: Remarkable atrophy occurs in benign hyperplastic prostatic gland after radical removal of the innervation of cholinergic parasympathetic pelvic nerve. Such operation may represent a novel therapy for BPH.

[The pathological change of rats' benign hyperplastic prostate after radical denervation].

OBJECTIVE: To study the pathological change of rats' benign hyperplastic prostate (BHP) after radical denervation. METHODS: A total of 65 male spontaneous hypertension rats (SHR) at 30 weeks age were randomly assigned into treatment group, sham surgery control group and normal control group. In surgery group, all the axonal branches of the major pelvic ganglion (MPG) supplying the bilateral prostate were truncated, followed performing of cystostomy; In sham surgery control group, only cystostomy was performed; In normal control group, no procedure was performed. The rats were sacrificed at 3, 7, 11, 15 and >or= 21 d post-operation respectively. The gross morphological changes of prostate in all animals were observed. RESULTS: In treatment group, the prostate in 3 d post-operation showed granular solidification and shrunken volume and the changes occurred gradually over time. The glandular epithelial cells showed gradual degeneration, necrosis and detachment. The glandular epithelium became progressively thinner, the smooth muscles elongated and thinned progressively and the stromal components showed mild to moderate overgrowth. At the later stage, the glandular epithelium, glandular lumen and smooth muscles gradually disappeared and the prostate was largely replaced by connective tissues. Electron microscopic study showed that the glandular cells gradually underwent vacuolar degeneration and the structures of basement membrane became fuzzy. The smooth muscles cells degenerated overtime and the fibroblasts and collagenous fibers in the stroma overgrew slowly. At the late stage, most of the glandular cells became necrotic, the basal membrane and smooth muscle cells disappeared and collagenous fibers were highly hyperplasic. In surgery group in 3 d post-operation, the S-100 staining of nerve fiber was diffuse and disappeared after 11 d while it persisted normally in other groups. The two values in sham surgery control group showed no significant changes post-operatively. CONCLUSIONS: After radical denervation, the rat prostate with benign hyperplasia (gland and smooth muscles) undergoes dramatic atrophic changes and the volume decreases significantly. It suggests that this treatment may represent a novel therapy for BPH.

Rapid identification of UCA1 as a very sensitive and specific unique marker for human bladder carcinoma.

PURPOSE: The most common genitourinary malignancy in China is bladder transitional cell carcinoma (TCC). Early diagnosis of new and recurrent bladder cancers, followed by timely treatment, will help decrease mortality. There are currently no satisfactory markers for bladder cancer available in clinics. Better diagnostic methods are highly demanded. EXPERIMENTAL DESIGN: In this research, we have used comprehensive expressed sequence tag analysis, serial analysis of gene expression, and microarray analysis and quickly discovered a candidate marker, urothelial carcinoma associated 1 (UCA1). The UCA1 gene was characterized and its performance as a urine marker was analyzed by reverse transcription-PCR with urine sediments. A total of 212 individuals were included in this study, 94 having bladder cancers, 33 ureter/pelvic cancers, and 85 normal and other urinary tract disease controls. RESULTS: UCA1 was identified as a novel noncoding RNA gene dramatically up-regulated in TCC and it is the most TCC-specific gene yet identified. The full-length cDNA was 1,439 bp, and sequence analysis showed that it belonged to the human endogenous retrovirus H family. Clinical tests showed that UCA1 assay was highly specific (91.8%, 78 of 85) and very sensitive (80.9%, 76 of 94) in the diagnosis of bladder cancer and was especially valuable for superficial G2-G3 patients (sensitivity 91.1%, 41 of 45). It showed excellent differential diagnostic performance in various urinary tract diseases without TCC. CONCLUSIONS: UCA1 is a very sensitive and specific unique marker for bladder cancer. It could have important implications in postoperative noninvasive follow-up. This research also highlights a shortcut to new cancer diagnostic assays through integration of in silico isolation methods with translational clinical tests based on RNA detection protocols.