Objective To study the influence of inhibited steroidogenic factor-1 on human adrenocortical H295R cells, and explore its role in the pathogenesis of adrenal tumors. Methods The plasmids pGenesil1-SF-1-shRNA which containing U6 promoter and SF-1-specific short hairpin RNA (shRNA) and pGenesil1-negative-shRNA containing unspecific shRNA were transfected into H295R cell. The expression of SF-1 was measured by Western blot and real-time polymerase chain reaction(RT-PCR). Cell proliferation was analyzed by WST-1 assay and cell count. Ki-67 expression was detected by immunohistochemistry and cell apoptosis was examined by TUNEL assay. Results Compared with those in control cells, the protein and mRNA level of SF-1- transfected cells were reduced by 69.7% and 71.2% (P＜0. 01). WST-1 and cell count method showed that SF-1 gene silencing obviously inhibited cell proliferation(P＜0. 01). By contrast, there was a 3. 7-fold increase in the percentage of apoptotic H295R cells in SF-1-inhibited group than that of control group (P＜0. 01). Immunohistochemistry showed that Ki-67 positive cells in SF-1-inhibited cells were lower than the negative control cells (16.90±2.17) % and (33. 48±3.16)%,(P＜0. 01). Conclusion SF-1 gene silencing can inhibit the proliferation of adrenocortical cells, and it is expected to become a key protein in understanding pathogenesis of adrenal tumors or treating them.

Objective Screening OSAHS patients on pregnancy with Modified Epworth Sleepiness Scale( Epworth sleepiness scale,ESS)and to assess its effect. Methods 22 patients underwent the pregnancy,and pregnancy with OSAHS( mild in 23 cases,22 cases of moderate and severe in 19 cases)group were 64 people, By ESS and modified ESS score, EP and modified EP values was derived. The neck circumference ( NC), body mass index (BMI) was measured. Conduct of polysomnography ( PSG), apnea hypopnea index ( AHI ) and lowest oxygen saturation ( LSaO2 )ESS and modified ESS correlation with the AHI was analyzed and ROC curves was drawn. Results The EP value of pregnancy with mild OSAHS group has no significant difference between normal pregnancy group( P ＞ 0.05) ;the rest of pregnancy OSAHS group modified EP, EP values and the normal pregnancy group were significantly different ( all P＜0.05) ;modified EP, EP, NC, BMI values positively correlated with the AHI value, the correlation coefficient r were :0.876,0.748,0.671,0.670 ( all P ＜ 0.001 ) ;modified EP, EP, NC, BM I of the A UC values were 0.901,0.819,0.750, 0.779; when the modified EP = 8.5, had higher sensitivity ( 84.4% ) and specificity ( 90.9% ).Conclusion Modified ESS on pregnancy OSAHS patients had better clinical value of screening.

Objective To evaluate the efficacy of self-retaining suture (QuillTM SRS) in retroperitoneal laparoscopic partial nephrectomy for complicated renal tumor by assessing perioperative parameters.Methods Between 2010 and 2012,78 cases of complicated renal tumor (R.E.N.A.L score ≥ 7) treated by retroperitoneal laparoscopic partial nephrectomy (LPN) with two layers continuous knotless barbed suture (QuillTM SRS group) (n=30) or traditional absorbable vicyl suture (non-SRS group) (n=48) were retrospectively analyzed.In QuillTM SRS group,2-0 Quill SRS was used to suture the deep wound bed,and the second outcr layer renorrhaphy was performed with a 1-0 Quill SRS by the same way.In non-SRS group,the inner layer was sutured using a 15cm in length 2-0 monicryl suture by the same method mentioned above.A second outer layer was sutured with 1-0 vicryl suture across the wound.Cases were matched for R.E.N.A.L score.Comparison was made in term of operation time,preoperative parameter and perioperative complications between SRS group and non-SRS group.Results Renorrhaphy was successfully performed in all cases except 1 case converting to open surgery in non-SRS group.Mean warm ischemia time in SRS group was shorter than non-SRS group (18 vs 25 min,P =0.021).The proportion of bleeding requiring intervention in the non-SRS group (7/48,14.5％) was 4.3-fold higher than that of the SRS group (1/30,3.3％),but the differernce is not significant (P＞0.05).There were no significant differences between two groups in postoperative creatinine changes.Limitations of this study include the absence of randomization and the relative small sample size.Conclusions SRS can be safely used for complicated renal tumor during LPN,and SRS can significantly reduce the WIT and may also reduce bleeding during the operation.

Objective To investigate the in vitro effects of bladder cancer cell proliferation after silencing Notch1 gene. Methods The siRNA eukaryotic expression vector of Notch1 (psiRNA1)was constructed and transfected into bladder cancer cell lines T24 and BIU-87. Methabensthiazuron (MTT) and flow cytometry (FCM) assays were used to detect bladder cancer cells line growth, cell cycle and apoptosis after the transfection. RT-PCR and Western blotting were used to determine the expression changes of Notch1 in these cell lines. Results After transfection for 72 h, the rate of G0/G1 phase cells inceased from (23.89±1.32) % to (80.13±2.69)% in T24 cell line, and increased from (24.63±1.68)% to (69.44±2.41)% in BIU-87 cell line (both P<0.05). In addition, apop-totic cell index in T24 and BIU-87 cell lines increased from (1.28±0.14)% to (13.75±1.23)%, from (1.01±0.27)% to (8.72±1.01)%, respectively(both P<0.05). The growth of T24 and BIU-87 cell lines was obviously inhibited 24 h after the transfection, and the inhibitory effects lasted until 96 h after the transfection. Notch1 mRNA and protein significantly downregulated after transfection compared to the control(P<0.05). Conclusions Silencing Notch1 expression can inhibit the prolif-eration of bladder cancer cell lines. Notch1 gene might act as a tumor gene in bladder cancer.

Objective To evaluate the efficacy and feasibility of laparoscopie intervention for congenital obstructive megaureter in children. Methods Eleven children with congenital obstructive megaureter(left in 4,right in 7)underwent laparoseopie ureteroplasty.One had congenital ureter oririce stenosis,9 had been diagnosed as simple congenital ureter orifice stricture,1 had recurrent ureter orifice stricture after open ureterovesical reimplantation.B-ultrasound and IVU showed severe hydronephrosis in 7 cases and moderate in 4. Results The operation was successful in all cases and none had urine leakage.The mean operating time was 103.0±35.3 min(range 70-190 min).The mean blood loss was 18.0±9.5 ml(range 10-40 ml)and the mean postoperative hospital stay was 8.0±1.4 d(range 7-10 days).The double J stent was removed 6 weeks after operation.The patients were followed up for 3-24 months(mean,6 months).Cystography showed no reflux in all cases during follow-up. Conclusion Laparoscopical ureteroplasty could be a minimal invasive,less suffering technique for the treatment of congenital obstructive megaureter in children.

MicroRNAs (miRNAs or miRs) are a class of short, non-coding RNAs that participate in various oncological processes. This study aims to explore the roles of microRNA-34a (miR-34a) in invasive urothelial bladder carcinoma. miR-34a was transfected into bladder cancer cell lines 253J and J82. The miR-34a expression levels in tissues and cells were detected by using qRT-PCR. The Notch1 expression was detected by qRT-PCR and Western blotting. Cell migratory and invasive abilities were measured by Transwell chamber assay. Bioinformatics and luciferase assay were performed to predict and analyze the binding sites between miRNA-34a and Notch1. It was found that there was aberrant expression of miR-34a in bladder cancer tissues. Moreover, we revealed that ectopic expression of miR-34a suppressed cell migration and invasion, while forced expression of Notch1 increased cell migratory and invasive abilities. Finally, we observed that miR-34a transfection significantly down-regulated luciferase activity and reduced the mRNA and protein levels of Notch1. Our study concluded that microRNA-34a antagonizes Notch1 and inhibits cell migration and invasion of bladder cancer cells, which indicates the tumor-suppressive function of microRNA-34a in bladder cancer.

OBJECTIVETo explore the relationship of continuous positive airway pressure and pulmonary function in patients with obstructive sleep apnea hypopnea syndrome (OSAHS), and formulate the prediction equation for the effective therapeutic pressure.

METHODSIn a retrospective cross-sectional study of 48 patients with established OSAHS, all the patients were carefully examined for their medical history, and overnight sleep monitoring was carried out to measure the sleep apnea-hypopnea index, mean oxygen saturation, minimum oxygen saturation, and oxygen drop index. The data of manual pressure titration (effective pressure) and pulmonary function tests (tidal volume, one second forced expiratory volume, central airway resistance, and peripheral airway resistance were collected for multiple linear regression analysis.

RESULTSThe effective therapeutic pressure was not correlated with the indices of the pulmonary functions in the patients, but showed correlations with the neck circumference, abdominal circumference, apnea-hypopnea index, mean oxygen saturation, least oxygen saturation, and oxygen desaturation index. Multiple linear regression (α=0.05 test level) identified only oxygen desaturation index (P=0.012) and mean oxygen saturation (P=0.036) as the dependent variables of the effective therapeutic pressure. Linear regression analysis showed that the effective therapeutic pressure had a linear relationship with the oxygen drop index and mean oxygen saturation, and was inversely correlated with the mean oxygen saturation.

CONCLUSIONSIn patients with OSAHS, the pulmonary function indices are not correlated with the effective therapeutic pressure. In the absence of manual pressure titration, the effective therapeutic pressure can be predicted using the prediction equation: effective therapy pressure = 24.262+0.044×oxygen desaturation index -0.19×average oxygen saturation.

Adult ; Aged ; Continuous Positive Airway Pressure ; Forced Expiratory Volume ; Humans ; Middle Aged ; Polysomnography ; Respiratory Function Tests ; Retrospective Studies ; Sleep Apnea, Obstructive ; physiopathology ; therapy

Objective To study the role of fibulin-5 in urothelial carcinoma of bladder. Methods Fibulin-5 expression was detected in bladder cancer tissues (13 cases of G_1 and G_2, 7 cases of G_3) and normal bladder mucosa samples by Western blotting assay. Fibulin-5 cDNA was amplified by PCR and cloned into pMD-19T simple vector. The pMD-19T-Fibulin-5 vector was digested by restriction endonucleases XhoI and EcoRI to generate a XhoI-Fibulin5-EcoRI fragment that was then ligated into the identical sites in p-EGFP-Nl plasmid to synthesize p-EGFP-Fibulin-5 plasmid. The p-EGFP-Fibulin-5 plasmid was finally transfected into bladder cancer cell line 5637. The migration and invasion of untransfected, vector-transfected and fibulin-5-transfected bladder cancer cells were measured by Boyden chamber assay. Results Compared to 1. 16 ±0. 28 in the normal control, the expression of fibulin-5 protein in low grade and high grade tumors were 0. 57±0. 32 and 0. 44±0. 42(P<0. 01, respectively). However, the difference between low grade and high grade tumors was not statistically significant (P>0. 05). The successfully transfected bladder cancer cells demonstrated green fluorescent light. The migrated cell number of fibulin-5-transfected cells was 127. 6 ± 3. 1 compared with 139. 3±7. 7 for vector-transfected cells and 136. 9±5. 7 for untransfected cells (P>0. 05, respectively). In contrast, the invaded cell number of fibulin-5-transfected cells was 8. 0±3. 1 compared with 31. 5±4. 8 for vector-transfected cells and 31. 7±4. 7 for untransfected cells (P<0. 01, respectively). Conclusion Fibulin-5 is down-regulated in urothelial carcinoma of bladder and acts as a tumor suppressor gene by inhibiting the invasion of bladder cancer cells.

Objective To develop a staged laparoscopic training program for performing the ana-tomic retroperitoneoscopic adrenalectomy(ARA), and to determine its safety and feasibility. Me-thods Five young urological doctors without previous experience in open adrenalectomy were selected third period, trainees acted as camera holder first, then performed simple operations such as laparo-scopic renal cyst unroofing. Finally, they performed 30 ARA independently under the mentor's super-vision. Pheochromocytoma was ruled out for its large tumor size and potential cardiovascular risk. The patient selection criteria were the same as those of the initial 30 cases performed by the tutor. Preope-rative data of the initial 30 ARA performed by each trainee and tutor which included gender, age, body mass index, tumor size, tumor location and pathological diagnosis of tumor were compared between trainees and the tutor. The intraoperative and postoperative data of 150 ARA in the trainees were compared with the initial 30 ARA of the tutor. These included mean operative time, estimated blood loss, length of hospital stay, conversion rate, complication rate. Qualitative and quantitative data were compared between the groups using x2 and t test statistics methods by SPSS 12.0 for Windows, except operative time, which was from a nonnormal distribution. A P value less than 0.05 was consi-dered to be statistically significant. Results Preoperative data of the initial 30 ARA performed by each trainee were marched to those of the mentor (all P>0.05). All ARA were completed successful-ly. No procedure converted to open surgery. The median operative time of the trainees was 82 min (range 59-133 min), which was less than that of the tutor [132 min (range 73-230 min), P< 0.01]. And the trainees' learning curve was flatter than their tutor's. Estimated blood loss and length of hospital stay for the 5 trainees and the tutor were 62.2±22.0 ml, 4.8±1.3 d and 63.9±21.1 ml, 4.5±1.4 d respectively. There was no significant difference between these results (both P>0.05). No major complication was observed. Though the total perioperative complication rates were no diffe-rence between the trainees and their tutor (8.0% versus 13.3%, P>0.05), intraoperative minor complication rates of the trainees (1.3%) was less than that of the tutor (10.0%, P<0.05). Con-clusion The staged laparoscopic training is safe and feasible for young urological doctor to study in performing ARA.

Objective To study the endoscopic anatomical structures in retroperitoneal space and to share experiences of retroperitoneoscopic radical nephrectomy. Methods Between January 2006 and March 2008, a total of 85 patients underwent retroperitoneoscopic radical nephrectomy. Thirty-eight tumors were on the left kidney and 47 on the right side. The mean tumor size was 5.5± 1.7 cm in diameter (2.5 to 10.5 cm). There were 74 cases in clinical stage T1N0M0 and 11 cases in T2N0M0. Following the principle of radical nephrectomy outside the renal fascia, the whole surgical procedure was performed along "2 spaces" and "2 poles". The ventral attachment of the kidney was dissected in anterior pararenal space between peritoneum and anterior renal fascia. The dorsal attachment was dissected in anterior psoas space between posterior renal fascia and psoas fascia. The cepha-lic attachment was dissected up to the subdiaphragmatic and down to iliac fosse. During the proce-dure, important anatomic structures such as parietal peritoneum and its reflexion, anterior renal fasci-a, lateroeonal fascia, posterior renal fascia, psoas muscles, greatvessels and their branches were care-fully identified. Results One case was converted to open surgery because of severe and extensive ad-hesion of the right kidney to the adjacent tissues. The other 84 procedures were successfully comple-ted. The median operative time was 65 rain (range 50 to 165 min) and median estimated blood loss was 58 ml (range 25 to 600 ml). Of all operations, peritoneum perforation occurred in 5 cases and small vessel injuries around renal pedicles were observed in 6 cases. Major complication such as great vessel injury was not observed. Mean follow-up of all 85 patients was 10 months (range 2 to 25 months). No local recurrence and port site tumor seeding was found. Conclusion During retrope-ritoneoscopic radical nephrectomy, studying anatomical features of renal area and recognizing impor-tant anatomic structures will help to improve the safety of the surgery and reduce morbidities.