[Effect of androgen receptor on IgG expression, proliferation and migration of prostate cancer cells in vitro].

OBJECTIVE: To investigate the effect of androgen receptor (AR) on IgG protein expression and the proliferation and migration of prostate cancer cells. METHODS: Western blotting was used to detect the expression of AR protein and IgG in androgen-dependent prostate cancer LNCap cells and castration-resistant prostate cancer PC-3 cells. In AR-overexpressing cells (PC-3-AR cells) established by transfecting PC-3 with AR gene (pCDNA3.1) and LNCap cells with small interfering RNA-mediated AR silencing (LNCap-siAR cells) were analyzed for expressions of AR protein and IgG with Western blotting; the expression of IgG mRNA was detected by Q-PCR, and the cell proliferation and migration were assessed with MTT assay and wound healing assay, respectively. RESULTS: Compared with PC-3 cells, LNCap cells expressed a higher level of AR protein and a lower level of IgG (P<0.05). PC-3-AR cells showed attenuated proliferation and migration with a lowered expression of IgG (P<0.01), while LNCap-siAR cells showed enhanced proliferation and migration with increased expression of IgG (P<0.01). CONCLUSION: The expression of AR is inversely correlated with IgG and is associated with the proliferation and migration of prostate cancer cells in vitro.

[Effect of sodium phenylbutyrate on the sensitivity of PC3/DTX-resistant prostate cancer cells to docetaxel].

OBJECTIVE: To investigate the effect of sodium phenylbutyrate (SPB) in modulating docetaxel resistance in human prostate cancer cells in vitro. METHODS: A PC3/docetaxel-resistant human prostate cancer cell line PC3/DTX was induced and examined for proliferation, viability, and cell inhibition rate in the presence of SPB. The concentration of concentration of docetaxel required to kill 50% of PC3/DTX cells incubated with 0, 1, 2, and 4 mmol/L SPB was determined using MTT assay. Cell apoptosis rate was analyzed with flow cytometry and the cellular expressions of p21, cyclin D1 and survivin proteins were detected using Western blotting. RESULTS: Treatment of PC3/DTX cells with 0, 1, 2, and 4 mmol/L of SPB for 48 h resulted in cell viabilities of (99.85∓2.69)%, (84.68∓3.87)%, (68.65∓4.54)% and (43.54∓5.69)%, and cell inhibition rates of (10.69∓3.65)%, (25.78∓4.58)%, (54.68∓3.98)% and (69.84∓6.54)%, respectively (P<0.05). The concentration of docetaxel required to kill 50% of PC3/DTX cells cultured in the presence of with 0, 1, 2, and 4 mmol/L SPB was 135.98∓2.69, 109.65∓3.87, 87.65∓3.84 and 64.62∓2.98 nmol/L, respectively (P<0.05), and the cell apoptosis rates were (7.2∓0.8)%, (10.2∓0.9)%, (19.8∓2.1)% and (27.4∓2.5)%, respectively. SPB treatment promoted the protein expression of p21 and suppressed the expressions of cyclin D1 and survivin in PC3/DTX cells. CONCLUSION: SPB can affect the expressions of p21, cyclin D1, and survivin in PC3/DTX cells and increase the sensitivity to the drug-resistant cells to docetaxel.

Can the lower urinary tract storage symptoms be completely resolved after plasmakinetic enucleation of the prostate?

The aim of this study was to determine whether the lower urinary tract storage symptoms of benign prostatic obstruction (BPO) could be completely resolved after plasmakinetic enucleation of the prostate (PKEP) and the possible predictors of persistent symptoms. Two hundred and sixty-seven cases of BPO performed PKEP from July 2008 to June 2009 were retrospectively analyzed. Five-year postoperative data were collected and compared with the preoperative data. According to the urodynamic results, the patients were divided into involuntary detrusor contraction (IDC) group (n = 95) and no IDC group (n = 172) preoperatively; the patients with IDC were divided into IDC-persistent group (n = 33) and IDC-resolved group (n = 62) after PKEP. The predictors of persistent IDC were analyzed. Compared with the preoperative data, the 5-year postoperative data showed that the IDC rate was lower (P = 0.000), Overactive Bladder Symptom Score (OABSS) was lower (P = 0.000), maximum cystometric capacity (MCC) was larger (P = 0.000), Prostate volume (PV) was smaller (P = 0.000), and prostate-specific antigen (PSA) was lower (P = 0.000). Compared with the no IDC group, the IDC group showed that the age was older (P = 0.016), MCC was smaller (P = 0.004), PSA was higher (P = 0.016), and Chronic Inflammation rate was higher (P = 0.004). Compared with IDC-resolved group after PKEP, IDC-persistent group showed that the age was older (P = 0.019), MCC was smaller (P = 0.000), PSA was higher (P = 0.013), and Chronic Inflammation rate was higher (P = 0.032). The present study shows that the storage symptoms are still needed to be focused on after PKEP. The advanced patient age, MCC, PSA, and chronic inflammation may be the important clinical predictors of persistent IDC.

Orthotopic detaenial sigmoid neobladder after radical cystectomy: technical considerations, complications and functional outcomes.

PURPOSE: In recent years the orthotopic neobladder has gained increasing popularity in patients who undergo radical cystectomy. However, there are only a few reports of orthotopic neobladders reconstructed from the sigmoid without detubularization. We investigated the complications and functional outcomes of the orthotopic sigmoid neobladder reconstructed using our detaenial technique. MATERIALS AND METHODS: We performed a retrospective study of the detaenial sigmoid neobladder in 210 consecutive patients who underwent radical cystectomy at our institution from January 2003 to March 2010. ANOVA was used to investigate urodynamic finding differentials with time. Univariable and multivariable analyses were done to determine factors influencing continence. RESULTS: Median followup was 48 months. Early complications (90 days or less) were observed in 65 patients (31%). Late complications (greater than 90 days) were observed in 45 patients (21.5%). Five-year daytime and nighttime complete continence rates were 74.6% and 57.1%, respectively. Younger age was the only independent factor associated with complete continence during the day (OR 2.342, 95% CI 1.803-3.041, p <0.001) and night (OR 1.193, 95% CI 1.087-1.310, p <0.001). Mean maximal capacity and post-void residual urine were 328.8 and 22.2 ml, respectively. The mean maximal flow rate was 18.5 ml per second. The mean end filling pressure, pressure at maximal capacity and maximal intravesical pressure were 35.8, 55 and 60.6 cm H2O, respectively. These parameters remained stable with time (each p >0.05). CONCLUSIONS: This study confirms that detaenial sigmoid neobladder is a safe, feasible alternative for urinary diversion.

Influence of reproductive tract obstruction on expression of epididymal proteins and their restoration after patency.

Vasectomy is a simple and reliable method of male contraception. A growing number of men after vasectomy request vasectomy reversal due to various reasons. The pregnancy rate is lower than the patency rate after vasovasostomy and the pregnancy rate is time dependent. In this study, we evaluated the influence of reproductive tract obstruction on expression of epididymal proteins and their restoration after patency. Adult male Wistar rats were studied 30, 60 and 120 days after vasectomy, 30 days after vasovasostomy or after sham operations. Two-dimensional gel electrophoresis, mass-spectrometric technique, multidatabase search, Western blotting and real-time PCR were used to analyze the expression regulation of epididymal proteins. Total integrated intensity and total spot area of autoradiograms showed a consistent downward trend with time after obstruction, and this trend remained after patency. The intensity of the autoradiographic spots in three patency groups showed three trends: a downward trend, similar intensity and an upward trend compared with the correspondent obstruction group, respectively. Further verified experiments on human epididymis 2 (HE2), fertilization antigen-1 (FA-1), clusterin and PH20 demonstrated that compared with the correspondent obstruction group, the translation levels of HE2 and the mRNA transcription levels of HE2 showed an upward trend in patency groups, especially in the groups of obstruction for 60 days where the expression levels of HE2 were significantly upregulated after patency (P<0.05). Reproductive tract obstruction provokes a disregulation of gene expression in the epididymis and this disregulation remained after patency. Successful reversal may recover some proteins and the recovery is time dependent. Obstruction differentially alters mRNA transcription of different proteins and the content of proteins seemed to be easier to be influenced than the gene transcription.

[Laparoendoscopic single-site surgery for male pseudohermaphroditism: the initial report].

OBJECTIVE: To evaluate the therapeutic effect of laparoendoscopic single-site surgery (LESS) for treatment of male pseudohermaphroditism. METHODS: A 17-year-old patient with male pseudohermaphroditism and a female social sex was admitted. According to the request by the patient and the relatives for a female gender, LESS vaginoplasty and cryptorchidectomy were performed using a single multilumen port inserted through a 2.5 cm incision below the umbilicus, followed by reconstruction of the perineal region by open surgery. RESULTS: The total operative time was 7 h, and the LESS procedure lasted for about 3.5 h. No other port incision was needed. The estimated intraoperative blood loss was 400 ml. No electrolyte or metabolic acid-base balance disorders were observed perioperatively. In the follow-up examination at 6 months after the operation, the reconstructed vagina healed smoothly without obvious contraction or fixation failure, and the perineal region showed good appearance. CONCLUSION: With minimal invasiveness, LESS surgery produces good cosmetic effect and allows rapid postoperative recovery, thus may become a promising alternative to the management of pseudohermaphroditism.

[Transurethral enucleation of the prostate for treatment of benign prostatic hyperplasia in patients less than 50 years old].

OBJECTIVE: To evaluate the therapeutic effect of transurethral enucleation of the prostate for treatment of benign prostatic hyperplasia in patients below 50 years of age. METHODS: Twelve patients with benign prostatic hyperplasia patients (mean age 48.2 years, range 46-49 years) underwent transurethral enucleation of the prostate. The middle lobe and two lateral lobes were enucleated with the preprosthetic sphincter and anterior fibromuscular stroma preserved during the operation. The patients were followed up to evaluate the lower urinary tract symptoms and sexual activity after the surgery. RESULTS: The 12 patients were followed up for 3 to 6 months. The symptoms of lower urinary tract obstruction were improved obviously after the surgery, and the International Prostate Symptom Score (IPSS) decreased from 24±5.1 to 8.8±1.4 and peak urine flow rate (Qmax) increased from 8.1±4.2 ml/s to 20.1±4.2 ml/s at 3 months postoperatively. All the 12 cases had residual urine (12-44 ml) preoperatively, but after the surgery, only 4 still had residual urine of less than 30 ml. All the patients had normal erection function postoperatively, and 10 had normal ejaculation; the other 2 patients recovered normal ejaculation 3 and 5 months after the operation, respectively. CONCLUSIONS: Transurethral enucleation can alleviate the low urinary tract obstruction symptom and improve the sexual function by avoiding preprosthetic sphincter injury in relatively young patients with benign prostatic hyperplasia.

[Laparoendoscopic single-site surgery radical cystectomy with orthotopic taenia myectomy sigmoid neobladder: initial report].

OBJECTIVE: To report the first case and detailed techniques of laparoendoscopic single-site surgery (LESS) radical cystectomy with orthotopic taenia myectomy sigmoid neobladder for organ-confined bladder cancer. METHODS: A 74-year-old man presented with gross hematuria for 2 months and biopsy revealed bladder cancer. LESS radical cystectomy and bilateral pelvic lymphadenectomies were performed using a single multilumen port inserted through a solitary 3.5 cm lower abdominal incision with conventional laparoscopic instruments. The taenia myectomy sigmoid pouch was then constructed by open procedure. RESULTS: The total operative time was 9.5 h, and the LESS procedure lasted for about 5.5 h. No other port incision was added. The final pathology revealed urothelial carcinoma. The estimated intraoperative blood loss was 600 ml with blood transfusion of 400 ml. The pelvic lymph nodes and the surgical margins of the ureters and urethra were all free of tumor invasion. No water electrolyte and metabolic acid-base balance disorders were observed perioperatively. The neobladder capacity was about 280 ml, with a residual urine volume of 10 ml and peak flow rate of 11.1 ml/s 3 months postoperatively. CONCLUSION: Although with a steep learning curve, LESS surgery can be a less invasive and promising alternative to muscle-invasive bladder carcinoma.

[Expression of special AT-rich sequence-binding protein in bladder urothelial carcinoma and its clinical significance].

OBJECTIVE: To detect the expression of special AT-rich sequence-binding protein (SATB1) in bladder urothelial carcinoma and investigate its correlation to the biological behavior of the carcinoma. METHODS: The expression of SATB1 mRNA was detected in 34 cases of bladder urothelial carcinoma and 14 normal bladder tissues by RT-PCR, and the protein expression of SATB1 was detected in 68 cases of bladder urothelial carcinoma and 17 normal bladder tissues by immunohistochemistry. The correlation between SATB1 expressions and the biological behavior of the tumor was analyzed. RESULTS: The expression of SATB1 was significantly higher in bladder urothelial carcinoma tissues than in normal bladder tissues (P<0.05). and the expression of SATB1 in the tumor tissues was correlated to the clinical stage and metastasis of the tumor. CONCLUSION: SATB1 expression can be associated with the development and metastasis of bladder urothelial carcinoma and may potentially serve as an indicator for predicting the prognosis of bladder urothelial carcinoma.

[Temperature dependent Raman spectra and micro-structure study of cuspidine in solid and liquid phases].

Cuspidine plays an important role in conventional metallurgical continuous casting mould flux. An UV laser source was used to record its ambient and high temperature Raman spectra (temperature range: 298-1 723 K) combined with a charge coupled device (CCD) detector. Both increasing and decreasing processes as well as characteristic spectra and shifts in wavenumber were observed. Micro-structure of cuspidine in liquid state is not unitary and different from that in solid state, suggesting multi clusters coexisting. Density functional theory (DFT) simulation method was applied to calculate its wavenumbers of Raman active vibrations by introducing the crystal spatial configuration model of cuspidine. Thus the experimental vibrational wavenumbers of the characteristic peaks could be assigned. This will help study physical and chemical behavior of cuspidine in continuous casting mould flux and provide an unique in-situ method under varying temperature with Raman spectroscopic technique.

[Reversion transcriptional expression of DAPK in bladder cancer T24 cells 5-aza-2'-deoxycytidine].

OBJECTIVE: To investigate the methylation status of the promoter of resion death associated protein kinase (DAPK) gene in bladder cancer cell (T24), and study the effect of 5-aza-2'-deoxycytidine (5-aza-dc) on DAPK gene reactive expression in T24 and its inhibitory effect on T24. METHODS: The bladder cancer cell T24 was treated with different doses of 5-aza-dc. The inhibitory effect and apoptosis rate were detected by MTT and flow cytometry, and the changes of DAPK mRNA and protein expression and the methylation status of DAPK promoter were assessed by RT-PCR, Western blotting, and methylation specific PCR, respectively. RESULTS: The growth of bladder cancer cell was inhibited significantly and the maximal apoptosis rate detected by flow cytometry was (24.12-/+1.4)%. DAPK mRNA was not expressed in bladder cancer cell T24 in normal conditions. DAPK mRNA and protein re-expressed after 5-aza-dc (12.5 micromol/L) treatment in cell line T24 for 24 h, and DAPK promoter became unmethylated. CONCLUSIONS: The promoter methylation can be an important factor for silencing the expression of DAPK in bladder cancer cell. 5-aza-dc can inhibit the growth and induce apoptosis of bladder cancer cells through reversing unmethylation status of DAPK promoter.

[Preparation of whole-kidney acellular matrix in rats by perfusion].

OBJECTIVE: To prepare rat whole-kidney acellular matrix (ACM) scaffolds using fluid perfusion method. METHODS: The kidneys with ureters and renal vessels were harvested from 12-week-old Wistar rats. Intravenous catheters were inserted through the renal arteries to establish channels for whole-kidney retrograde perfusion successively with heparinized PBS, 1% SDS, deionized water, 1% TritonX-100 and antibiotic-containing PBS under a pressure of 100 cmH2O. After decellularization, the scaffolds were observed under microscope with HE staining, scanning electron microscope, and fluorescence microscope with DAPI fluorescence staining. RESULTS: No cell residue was found in the scaffolds under microscope. Scanning electron microscope identified reticular structures consisting of basilar membrane and collagen without normal cellular structures in the scaffolds, and no strong fluorescence due to the binding of DAPI to the cell nuclei was observed under fluorescence microscope. CONCLUSION: Fluid perfusion is simple and reliable to prepare rat whole-kidney acellular matrix, which may serve as an ideal cell-free scaffold.

[Laparoscopic radical cystectomy and detenial sigmoid colon orthotopic neobladder reconstruction for bladder tumor in a 3-year-old boy].

OBJECTIVE: To present a case of laparoscopic radical cystectomy and detenial sigmoid colon orthotopic neobladder reconstruction for bladder tumor in a child. METHODS: A 3-year-old boy with bladder rhabdomyosarcoma underwent laparoscopic radical cystectomy and detenial sigmoid colon orthotopic neobladder reconstruction. The bilateral pelvic lymphadenectomy and cystectomy were performed laparoscopically, and removal of the mobilized specimens and urinary diversion operation were managed through enlarged abdomen incision. The urinary diversion procedure included construction of the detenial sigmoid pouch, bilateral stented antiflux implantation of the ureters in the pouch and orthotopic anastomosis of the neobladder with the urethra. RESULTS: The total operative time was 6 h, and the laparoscopic procedure lasted for about 3.5 h. The intraoperative blood loss was 50 ml, and 200 ml concentrated red blood cell transfusion was used for the safety of the patient. Six dissected lymph nodes in each pelvic side and the surgical margins of the ureter and urethra were all free of tumor invasion. Bowel peristalsis recovered 3 days after the operation, and the pelvic drainage and the neobladder drainage tubes were removed on day 7 and 14, respectively. The urethral catheter and ureteral stents were removed 25 days after the operation. The daytime urine control and micturition recovered 1 week after the operation. The neobladder capacity was about 110 ml, with residual urine volume of 10 ml and peak flow rate of 12 ml/s after 5 months. No perioperative complications occurred such as water-electrolyte and metabolic acid-base balance disorders, urinary leakage, reflux or bowel obstruction. CONCLUSION: Laparoscopic radical cystectomy is minimally invasive, reduces intraoperative blood loss and allows rapid postoperative recovery, and can be a promising approach to management of bladder rhabdomyosarcoma in children.

[Cytocompatibility of silk fibroin film with rabbit urinary bladder transitional epithelial cells in vitro].

OBJECTIVE: To obtain large quantities of well differentiated urinary bladder transitional epithelial cells for used as the seed cells in bladder tissue engineering, and evaluate the cytocompatibility of silk fibroin film with the transitional cells in vitro to assess the possibility of tissue-engineered urinary organ construction. METHODS: The urinary bladder transitional epithelial cells were isolated from the bladders of New Zealand rabbits and cultured in vitro as the seed cells, whose morphology was observed and the specific protein (cytokeratin) expression identified by immunofluorescence assay. The cells were seeded in 96-well plates at 1 x 10(>4)/ml and incubated with silk fibroin film leaching solution or culture medium (negative control). MTT assay was performed to determine the cell proliferation rates of the wells and evaluate the cytotoxicity and cytocompatibility of the silk fibroin film. RESULTS: The isolated urinary bladder transitional epithelial cells reached confluence after 9-10 days of culture, which showed positive staining for immunocytochemistry with monoclonal antibody against cytokeratin. The absorbance of the cells culture in the presence of silk fibroin film leaching solution averaged 0.424-/+0.020, 0.996-/+0.118 and 1.285-/+0.048 after at 24, 72 and 120 h of cell culture, and that of the negative control group at the time points was 0.419-/+0.030, 1.105-/+0.098 and 1.228-/+0.052, respectively, showing no significant difference between the two groups (P>0.05). CONCLUSION: Silk fibroin film has good cytocompatibility with rabbit urinary bladder transitional epithelial cells, and may serve as good scaffold material for urologic tissue engineering.

[Structure and Raman spectra of titanium oxides].

Under the first-principle density functional theory, structures of several titanium oxides (TiO, Ti2O3, Ti3O5, anatase and rutile) were optimized, and the obtained the structure parameter that coincides with the X-ray crystal diffraction result. The optimized structure parameters coincide well with those obtained by X-ray diffraction method. Molecular vibrational modes were also studied and assigned. Double Numeric including D-polarization function basis set was used with local density approximation, and the local exchange-correlation energy was described with Vosko-Wilk-Nusair and spin non-restricted function. The calculated vibrational wavenumbers were used to interpret observed experimental Raman spectra of those titanium oxides. By comparing the Raman spectra of those titanium oxides with the calculated results, the characteristic peaks of various titanium oxides could be assigned. So the types of titanium oxides could be diagnosed and recognized. This will help to investigate the correlation between the structure and property of materials.

[Application of silk fibroin film for repairing rabbit urethral defect].

OBJECTIVE: To evaluate the effect of silk fibroin film for repairing urethral defect in rabbits. METHODS: Twenty-four male New Zealand rabbits were randomized into experimental group, control group I and control group II. In the experimental group, a urethral defect of 1.5 cm was induced in the 12 rabbits and repaired with silk fibroin film. The 6 rabbits in control group I without the surgically induced defect served as the sham operation group, and in control group II consisting of 6 rabbits the urethral defect of 1.5 cm was induced without repair. Histological observation and immunohistochemistry were conducted to examine the regenerative segments of the urethra at regular time points between 2 and 16 weeks postoperatively. RESULTS: The 12 rabbits in the experimental group did not show signs of urethral stricture following the surgery. The implanted silk fibroin film for defect repair was degraded completely at 16 weeks and the defect was repaired with smooth urethral mucous membrane lining and orderly arranged smooth muscle cells. Immunohistochemistry identified the cells lining the defect area as the urethral epithelial cells. CONCLUSION: Silk fibroin film can promote the repair of urethral defect by inducing the growth of the urethral epithelial cells and smooth muscle cells.

[Application of intracavitary retrograde dissection in transurethral vaporization resection of the prostate].

OBJECTIVE: To evaluate the effect of intracavitary retrograde dissection during transurethral vaporization resection of the prostate (TUVRP). METHODS: The efficacy of intracavitary retrograde dissection of the prostate with PK bipolar generation was evaluated in 260 cases in view of the volume of intraoperative bleeding, operation time, safety and postoperative bleeding, catheterization time and prognosis in comparison with the results of exclusive use of TUVRP. RESULTS: Intracavitary retrograde dissection of the prostate provided good operative view as compared with conventional TUVRP. CONCLUSION: Intracavitary retrograde dissection is a practically feasible procedure for benign prostatic hyperplasia.

[Laparoscopic radical cystectomy and sigmoid colon orthotopic neobladder reconstruction: report of 26 cases].

OBJECTIVE: To review our experience with intracorporeal laparoscopic radical cystectomy and sigmoid colon orthotopic neobladder reconstruction. METHODS: The clinical data of 26 cases of bladder carcinoma treated with the indicated surgical procedures were reviewed. RESULTS: The surgeries were successful in all the cases with the operating time ranging from 240 to 390 min, blood loss of 400 to 800 ml and red-cell transfusion of 0-4 U. Oral food intake was allowed 4-8 days after the operation, ureteral stents were removed in weeks 3 to 8 and the pouch catheter was removed in week 4 postoperatively. Daytime urinary continence was excellent and urinary incontinence at night occurred in 8 patients 3 months after the operation. CONCLUSION: Sigmoid colon orthotopic neobladder reconstruction can be effective for urinary diversion to ensure good quality of life of the patients.

[Sclerotherapy for renal cyst using erythromycin and procaine via renal puncture guided by B-ultrasound: report of 1,000 cases].

Our experience of sclerotherapy for renal cyst injection of erythromycin and procaine via ultrasound-guided renal puncture is reported. The total efficiency reached 96% with low recurrence rate in the 1,000 cases receiving this therapy, suggesting that this approach is a simple, accurate and effective renal cysts with minimized postoperative complications treatment.