Effect of stable DNA methyltransferase 3bknockdown on proliferation and apoptosis in bladder cancer cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of stable knockdown of DNA methyltransferase 3b (DNMT3b) on the proliferation and apoptosis of bladder cancer cells.</p><p><b>METHODS</b>Lentivirus expressing DNMT3b siRNA or the negative control siRNA was infected in human bladder cancer BIU-87 cells. MTT assay and flow cytometry were used to detect cell proliferation and apoptosis, respectively. The inhibitory effect of DNMT3b knockdown on xenograft tumors in nude mice was observed. Real-time PCR and Western blotting were carried out to investigate the expression level of cell apoptosis related genes. Methylation specific PCR was used to examine the methylation in the promoter region of the cell apoptosis related genes.</p><p><b>RESULTS</b>The results of real-time PCR and Western blotting showed that DNMT3b mRNA and protein level were stably knocked down in BIU-87 cells. Stable DNMT3b knockdown suppressed BIU-87 cell growth and the tumor formation ability of the cells in nude mice. DNMT3b knockdown promoted the apoptosis of BIU-87 cells, increased the mRNA and protein expression of the cell growth and apoptosis related genes including DAPK, Bax and RASSF1A, and significantly decreased the methylation of these genes.</p><p><b>CONCLUSION</b>Stable DNMT3b knockdown can affect the methylation of the cell growth and apoptosis related genes to regulate their expression, which might be a possible mechanism for suppressed cell growth and enhanced apoptosis of BIU-87 cells.</p>

Expression of myeloid-derived suppressor cells in the peripheral blood and its clinical significance in renal carcinoma / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of CD33⁺ HLA-DR⁻ myeloid-derived suppressor cells (MDSCs) in the peripheral blood of patients with renal carcinoma and its correlation with the clinicopathological features of renal cancer.</p><p><b>METHODS</b>Forty-four patients with renal carcinoma treated in our hospital between June, 2011 and October, 2012 and 18 healthy volunteers were enrolled in this study. Flow cytometry was performed to detect CD33⁺ HLA-DR⁻ MDSCs in the peripheral blood, and its correlation with the clinicopathological features of the patients were analyzed.</p><p><b>RESULTS</b>The positivity rate of CD33⁺ HLA-DR⁻ MDSCs in the peripheral blood was significantly higher in the cancer patients than in the healthy controls [(1.91 ± 0.66)% vs (0.62 ± 0.22)%, P<0.001]. The expression levels of CD33⁺ HLA-DR⁻ MDSCs in patients with renal carcinoma showed significant differences between stage I+II [(1.46 ± 0.44)%] and stage III [(2.04 ± 0.35)%] patients (P<0.01) and between stage III and stage IV patients [(2.50 ± 0.64)%] (P<0.05), but did not differ significantly in respect of age or gender.</p><p><b>CONCLUSION</b>CD33⁺ HLA-DR⁻ MDSCs expression in the peripheral blood is associated with tumor stage and differentiation in renal carcinoma and may play an important role in predicting the prognosis and tumor immunology of renal carcinoma.</p>

Expression of OCT4 protein in bladder cancer and its clinicopathological implications / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the expression of OCT4 protein in bladder cancer and its correlation to the clinicopathologic features and prognosis of bladder cancer.</p><p><b>METHODS</b>OCT4 mRNA and protein expression was detected in 5 bladder cancer cell lines (RT-4, Tcc-Sup, KK47, T24, and 5637) and 1 normal bladder cell lines by real-time PCR and Western blotting, respectively. Immunohistochemical analysis was used to detect the expression of OCT4 protein in 46 bladder cancer samples.</p><p><b>RESULTS</b>All the 5 bladder cancer cell lines expressed detectable levels of OCT4 mRNA and proteins, whereas the normal bladder cell line SV-HUC-1 was negative for OCT4 expression. The clinical bladder cancer tissues showed a high positivity rate of OCT4 expression (76.1%), which was not detected in normal bladder tissues. Specific OCT-4 signals were localized mainly in the nuclei of the cancer cells. The expression rate of OCT4 protein was significantly higher in bladder cancer tissue than in normal bladder epithelium (P<0.05), and showed a positive correlation to the grade of tumor differentiation and metastasis (P<0.05) but not to the patients' age, gender or TNM stage.</p><p><b>CONCLUSION</b>OCT4 protein expression is associated with tumor differentiation and metastasis in bladder cancer and may play an important role in the early diagnosis and prognostic evaluation of bladder cancer.</p>

Effect of docetaxel on C-jun and androgen receptor interaction in prostate cancer LNCaP cells and its androgen-independence subtype LNCaP-bic cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of docetaxel on the interaction between C-jun and androgen receptor (AR) in prostate cancer LNCaP cells and its androgen-independence subtype LNCaP-bic cells.</p><p><b>METHODS</b>LNCaP and LNCaP-bic cells were treated with docetaxel and the changes in AR and AP-1 expression were evaluated using luciferase assay. Western blotting and immunoprecipitation assay were employed to analyze the effects of docetaxel on the expressions of C-jun and AR and their interaction.</p><p><b>RESULTS</b>Luciferase assay showed that LNCaP and LNCaP-bic cells expressed higher levels of AR and C-jun after docetaxel treatment. Docetaxel induced a higher level of p-c-jun expression in LNCaP-bic cells than in the parental LNCaP cells. Western blotting showed a strong PSA protein expression in LNCaP-bic cell after docetaxel treatment. Docetaxel caused a stronger interaction between AR and C-jun in LNCaP-bic cells.</p><p><b>CONCLUSION</b>Docetaxel activates ligand-independent AR transcription, and the interaction between AR and C-jun may affect the outcome of docetaxel chemotherapy.</p>

Laparoendoscopic single-site retroperitoneal lymph node dissection: initial experience and 1-year follow-up / 南方医科大学学报

<p><b>OBJECTIVE</b>To summarize our initial experience with laparoendoscopic single-site (LESS) retroperitoneal lymph node dissection (RPLND) for treatment of nonseminomatous testicular cancer.</p><p><b>METHODS</b>From September 2010 to June 2011, 3 patients (aged 19-27 years) with right testicle enlargement and elevated alpha-fetoprotein level underwent right radical orchidectomy. Histopathological analysis revealed nonseminomatous germ cell tumor. LESS-RPLND was performed 3 weeks after orchiectomy. The homemade port was inserted through a 3-cm right pararectal incision in the right lower quadrant for unilateral RPLND using nerve-sparing technique and modified right-sided template removal similar to those in open RPLND.</p><p><b>RESULTS</b>The operation was successfully performed with a mean operative time of 240 min and a mean estimated blood loss of 50 ml. No conversion to open or conventional laparoscopic surgery was required. No major perioperative complications were observed. For the first case, the number of lymph nodes obtained for final histopathological examination was 11, and two positive nodes were detected. For the other 2 cases, no positive nodes were detected. Chemotherapy was administered in the first case. Alpha-fetoprotein level decreased close to the baseline one week postoperatively and no relapse occurred in these cases 3 month after RPLND. Follow-up at 1 year after the surgery showed good tumor control and preservation of the sexual function.</p><p><b>CONCLUSION</b>LESS-RPLND is safe and feasible for treatment of nonseminomatous testicular cancer, and the pararectal incision provides an ideal surgical approach with good cosmetic result, but the long-term effect needs to be tested by further large population-based study.</p>

Single-port laparoscopic surgery for radical nephrectomy: report of 22 cases / 南方医科大学学报

<p><b>OBJECTIVE</b>[corrected] To evaluate the method and technique of single-port laparoscopic radical nephrectomy.</p><p><b>METHODS</b>Form January 2009 to September 2011, 22 patients with renal carcinoma were treated with single-port laparoscopic radical nephrectomy. An incision about 5 cm in length was made through the umbilicus or in the postaxillary line under the 12th rib to establish the peritoneal or retroperitoneal working space. A single-port cannulation was deployed and the operation was carried out using standard and crooked laparoscopic equipment.</p><p><b>RESULTS</b>The operations were completed successfully in all the 22 cases without conversion to open surgery, but additional trocar was needed in 5 cases. The mean operative time of radical nephrectomy was 150 min (90-240 min). The mean postoperative hospital stay was 7.6 days (3-15 days). The operation left a roughly 5-cm-long scar in all the cases.</p><p><b>CONCLUSION</b>Single-port laparoscopic radical nephrectomy is safe and feasible with good cosmetic effect and shows much potential in radical resection of renal carcinoma.</p>

Pure laparoendoscopic single-site radical cystectomy: Initial experience with 10 cases report / 中华泌尿外科杂志

Objective To present our initial experience of pure laparoendoscopic single-site surgery (LESS) for radical cystectomy and bilateral pelvic lymph node dissections. Methods 10 patients with pathology confined bladder urothelial carcinoma underwent laparoendoscopic single-site radical cystectomy, including 9 males and 1 female. After a 3-4 cm lower median abdominal incision was made, quadport or homemade single multichannel port was inserted, and conventional and prebent laparoscopic instruments were utilized. The surgical procedure included bilateral pelvic lymphadenectomies, radical cystectomy and building with a sigmoid orthotopic neobladder by open surgery.Results No extra port needed, neither conversion to open or conventional laparoscopic surgery. The time of LESS procedure ranged from 130 to 330 min (mean 243 nin). Estimated blood loss ranged from 50 to 600 ml (mean 270 ml). 5 patients needed blood transfusion of 2 to 4 units. The pathologic evaluation revealed bladder urothelial carcinoma, negative margins and negative pelvic lymph node involvement. No mortality or severe complications were observed perioperatively. After followup of more than 6 months, all revealed controllable urination at daytime, while 4 revealed nocturnal incontinence and needed one or two pads during nighttime. No evidence of recurrent or metastatic disease was detected. Conclusions LESS radical cystectomy and bilateral lymphadenectomies was safe and feasible, and short-term follow-up showed good tumor control outcomes. Homemade single multichannel port made of two elastic ring and glove was simple and effective.

Cellular biocompatibility of whole-kidney acellular matrix in rats by perfusion / 中国组织工程研究

BACKGROUND: At present, there is little related report about producing a whole-kidney acellular matrix (ACM) scaffold in rats using perfusion. The cellular biocompatibility of the ACM is poorly understood. OBJECTIVE: To produce a whole-kidney ACM scaffold in rats by perfusion, to evaluate the cytocompatibility of ACM with the L929 cells in vitro, and to assess the possibility of ACM as the cytoskeleton and tissue-engineered urinary organ construction. DESIGN, TIME AND SETTING: An in vitro observation was performed at the Central Laboratory of Zhujiang Hospital, Southern Medical University from February to May 2009. MATERIALS: Kidneys were obtained from 12-week-old Whista rats, while ureter, renal veins and renal artery were reserved. Intravenous catheters were inserted through renal arteries to establish channels for perfusion. Whole-kidney retrograde perfusion was performed with successively heparinized PBS, 1% SDS, deionized water, 1% TritonX-100 and antibiotic-containing PBS under a pressure of 9.81 kPa to prepare whole-kindney acellular matrix scaffolds. METHODS: ① Samples were randomly divided into blank group (without any cells), negative control group (culture media), experimental group (rat kidney ACM leaching liquor), and positive control group (culture media containing 0.64% phenol). L929 cells in the logarithmic phase were seeded in 96-well plates at the density of 4×10~3/well, with 5 wells in each group. At 24, 72, and 120 hours after incubation, cells were stained with MTT method to detect absorbance at 490 nm and calculate relative growth rate. ② Control group (culture medium), experimental group (rat kidney ACM leaching liquor), and positive control group (culture media containing 0.64% phenol) were set up to detect cell apoptosis at 48 hours after culture using flow cytometry. MAIN OUTCOME MEASURES: Microstructure of the scaffold, cytotoxicity and cell apoptotic rate. RESULTS: After SDS and TritonX-100 union processing, reticulate structures made of basilar membrane and collagen were shown under scanning electron microscope rather than normal structures of cells. At every time intervals (24, 72, and 120 hours), there was no significant difference in the absorbance between experimental group and negative control group (P > 0.05). The grade of the cytotoxicity of the ACM was .0-1. There was no significant difference in cell apoptotic rate between experimental group and negative control group (P > 0.05). CONCLUSION: The whole-kidney acellular matrix scaffolds in rat by perfusion have good biocompatibility.

Construction of Detenial Sigmoid Neobladder after Laparoscopic Radical Cystectomy:Report of 71 Cases / 中国微创外科杂志

Objective To report our experience on construction of detenial sigmoid neobladder after laparoscopic radical cystectomy in 71 cases.Methods From August 2002 to May 2006,a total of 71 patients with invasive bladder carcinoma underwent construction of detenial sigmoid neobladder after laparoscopic radical cystectomy in our hospital.After the bladder was excised by laparoscopy,a 5-to 7-cm incision was made on the abdomen to remove the resected tissues,and then a 15-to 20-cm sigmoid colon was resected,the two colic bands opposite to the mesentery and the circular muscle and seromuscular layers between them were removed to construct a detenial sigmoid neobladder.Afterwards,the neobladder was anastomosed with the posterior urethra.Laparoscopic anastomosis was performed in 26 of the cases.Results The operation time was 240-390 min totally in the 71 cases.Laparoscopic radical cystectomy was finished in 80-270 min(mean,180 min),and the open surgery was completed in 160-240 min(mean 140 min).Oral intake was started at day 4-8 postoperation,ureteral stents were removed at week 3-4,and urethral catheter was removed at week 4.Three months after the operation,the mean intra-bladder pressure was 22.0 cm H2O(17-38 cm H2O),average bladder reservoir was 340 ml(200-410 ml),volume of residual urine was less than 25 ml in all the cases(