Objective:To investigate the experience and results of the modified lateral prostate capsule sparing robot-assisted radical cystectomy-orthotopic ileal neobladder (LPCS-RARC-OIN).Methods:From December 2018 to November 2020, 19 patients received LPCS-RARC-OIN by a single surgeon in Sun Yat-sen Memorial Hospital, Sun Yat-sen University. LPCS-RARC-OIN was performed on male patients with high-risk non-muscle-invasive bladder cancer or muscle-invasive bladder cancer cT 2N 0M 0 without tumour in the bladder neck or urethra, and prostate cancer was ruled out by MRI and serum PSA<2.5ng/ml. The average age was 57.6 years, the average IIEF-5 score was 20.4. Separating the prostatic adenoma and the lateral prostate capsule from the base to the apex of the prostate, and retaining the lateral prostate surgical capsule or lateral prostate capsule about 1-2mm thickness. Patients were followed up and urinary function, sexual function and oncological outcomes were recorded. Results:All 19 operations were finished successfully. The average operation time was 279.9 (225-345) min and average estimated blood loss was 88.9 (30-200) ml. The average postoperative hospital stays was 15.8 (9 -23) days. The average lymph node yields was 23.3 (11-42). All surgical margins were negative and no incidental prostate cancer was found. 2 weeks, 1 month, 3 months and 6 months after catheter removal, the daytime and nighttime continence were 42.1% (8/19)and 36.8% (7/19), 63.2% (12/19)and 63.2% (12/19), 78.9% (15/19) and 73.7% (14/19), 94.7% (18/19) and 89.5% (17/19), respectively. 3 months and 6 months after operation, the average IIEF-5 score was 7.2 and 10.1 points respectively. The average follow-up was 10.6 months (5.4-26.1 months)and no recurrence or distant metastasis was found in this study.Conclusions:LPCS-RARC-OIN could improve the urinary and sexual function in selected patients. However, the long-term follow up is needed for functional and oncological outcomes.

BACKGROUNDProstate specific membrane antigen (PSMA) can facilitate the growth, migration, and invasion of the LNCaP prostate cancer cell lines, but the underlying molecular mechanisms have not yet been clearly defined. Here, we investigated whether PSMA serves as a novel regulator of the phosphatidylinositol 3-kinase (PI3K)/Akt signaling by employing PSMA knockdown model and PI3K pharmacological inhibitor (LY294002) in LNCaP prostate cancer cells.

METHODSPSMA knockdown had been stably established by transfecting with lentivirus-mediated siRNA in our previous study. Then, LNCaP cells were divided into interference, non-interference, and blank groups. We first testified the efficacy of PSMA knockdown in our LNCaP cell line. Then, we compared the expression of PSMA and total/activated Akt by Western blotting in the above three groups with or without LY294002 treatment. Furthermore, immunocytochemistry was performed to confirm the changes of activated Akt (p-Akt, Ser473) in groups. Besides, cell proliferation, migration, and cell cycle were measured by CCK-8 assay, Transwell analysis, and Flow cytometry respectively.

RESULTSAfter PSMA knockdown, the level of p-Akt (Ser473) but not of total-Akt (Akt1/2) was significantly decreased when compared with the non-interference and blank groups. However, LY294002 administration significantly reduced the expression of p-Akt (Ser473) in all the three groups. The results of immunocytochemistry further confirmed that PSMA knockdown or LY294002 treatment was associated with p-Akt (Ser473) down-regulation. Decrease of cell proliferation, migration, and survival were also observed upon PSMA knockdown and LY294002 treatment.

CONCLUSIONSTaken together, our results reveal that PI3K/Akt signaling pathway inhibition may serve as a novel molecular mechanism in LNCaP prostate cancer cells of PSMA knockdown and suggest that Akt (Ser473) may play a critical role as a downstream signaling target effector of PSMA in this cellular model.

Antigens, Surface ; genetics ; metabolism ; Cell Line, Tumor ; Glutamate Carboxypeptidase II ; genetics ; metabolism ; Humans ; Male ; Phosphatidylinositol 3-Kinases ; metabolism ; Prostatic Neoplasms ; enzymology ; genetics ; therapy ; Proto-Oncogene Proteins c-akt ; metabolism ; RNA Interference ; Signal Transduction ; genetics ; physiology

BACKGROUND: Prostate cancer stem cell is an important reason for the invasion and recurrence of prostatic carcinoma. However, separation efficiency of prostate cancer stem cells is very low.OBJECTIVE: To explore the efficient method for isolating and identifying the prostate cancer stem cells from human prostatic carcinoma cell lines PC-3 and LNCap. METHODS: Prostate cancer cell lines PC-3 and LNCap were cultured in serum free medium (SFM) and serum supplemented medium (SSM), respectively. The percentage of prostate cancer stem cells from different medium was detected by flow cytometry through markers CD133 and CD44, and the properties of prostate cancer stem cells were preliminarily identified using inducing differentiation experiments.RESULTS AND CONCLUSION: PC-3 and LNCap formed sphere cells in SFM, which can be induced into adherent cells after culture in SSM. Higher percentage of CD44+/CD133+cells was obtained from LNCap cells (1.71%; 0.73%) than PC-3 cells (0.59%; 0.32%) in both SFM and SSM. The number of CD44+/CD133+ LNCap cells was more than PC-3 using both methods (P < 0.05), but the efficiency of SFM and SSM had no statistical significance (P > 0.05). However, the culture cycle was longer and number of obtained cells was less by SFM culture, directly influencing functional determination of prostate cancer stem cells. Compared with suspension culture method with SFM, SSM is more convenient and effective in isolating prostate cancer stem cells from LNCap cells.

Objective To present our initial experience in laparoscopic radical prostatectomy performed through an umbilical incision using a home-made multichannel port. Methods From August 2009 to March 2010, we performed single-port laparoscopic radical prostatectomy in 11 patients with localized prostate cancer. A home-made multichannel port was inserted extraperitoneally through a 3-cm umbilical incision. The single port extraperitoneal procedures included obturator fossa lymphadenectomy, radical prostatectomy and urethro-vesical anastomosis, while the urethro-vesical anastomosis was performed by a slip-knot running suture technique. Data were collected and analyzed prospectively. Results All cases were completed successfully, without conversion to a standard laparoscopic approach or open surgery except adding an additional port in one case. The average operative time was 256 minutes (range195-315), and the mean blood loss was 90 ml (range 20- 180), without any blood transfusion. The postoperative hospital stay was 15.4 days (range13- 24), and the Foley catheter was removed 12 days after surgery. No intraoperative complications occurred. One patient developed a vesico-rethralanastomosis leakage, 2 had lymphatic leakage and 1 had urinary tract infection,all of the cases were managed successfully with conservative treatment. Histopathological results showed negative surgical margine and negative lymph node dissection. All patients had no biochemical relapse after an average follow-up of 7 months. Conclusions Single-port laparoscopic radical prosta tectomy is feasible, cosmetic and minimally invasive with a low complication rate and good short-term outcome. Additional investigation is needed to evaluate the long-term safety and oncologic adequacy of this new approach.

Objective To evaluate the long-term results of minimal incision osteotomy for hallux valgus.Methods From February 1996 to May 1999,372 cases(705 feet)with hallux valgus were treated with minimal incision osteotomy.Seventy-nine cases after surgery were followed up for more than 5 years,including 6 males(10 feet)and 73 females(140 feet)with an average age of 47 years(range,13-75 years).The average follow-up time was 7.5 years(range,5.3-13.2 years).The preoperative and postoperative hallux valgus angles(HVA),intermetatarsal angles(IMA),the tibial sesamoid position(TSP),American orthopedic foot and ankle society(AOFAS)score,range of movement of the first metatarsophalangeal joint,and lateral metatarsalgia etc.were observed and measured.And the clinic outcome were studied and evaluated.Results Based on Wen Jian-min's clinic curative effect evaluation criterion(2001),56 feet(37.3%)were excellent,88 feet(58.7%)good ,6 feet(4.0%)fair.The rate of excellent or good was 96.0%(144/150).The postoperative mean AOFAS score was 84.20±4.32 points(range,60-100 points).The mean HVA decreased from 33.28 to 12.31 degrees,the mean IMA from 11.75 to 6.80 degrees.The TSP was corrected from an average preoperative grade of 4.29 to a grade of 3.07 at final follow-up.There are no nonunion or delayed union of osteotomy,avascular necrosis of the 1st metatarsal head,infection,hallux varus.Four feet(2.7%)had numbness in the medial of the big toe.The 1st metatarsophalangeal joint range of motion decreased from 70.20 to 69.53 degrees.There were 97 feet(64.7%)with the 2-5 metatarsalgia before operation,and 35 feet(23.3%)disappeared,54 feet(36.0%)improved,8 feet(5.3%)aggravated after operation.Conclusion Minimal incision osteotomy technique is sound and reliable method to treat hallux valgus.The osteotomy technique is simple,and could provide satisfactory long-term results with lower complication rate.

Objective To verify the efficiency of a new laparoscopic urethrovesical anastomosis training model by comparing it with the chicken skin model. Methods Chicken posterior trunks and porcine colons were used to construct the training model. The posterior trunk of a chicken was used to simulate a human pelvis, and a 3-mm cloacal stump was used to simulate a human urethral stump. A 15-cm segment of porcine colon with a 1-cm orifice was used to simulate a human bladder or neobladder. An imitation urethrovesical anastomosis was performed with laparoscopic instruments in a laparoscopic training box. Forty trainees with no laparoscopic experience were randomized into 2 groups.The trainees in group A (n=20) practiced using this new model for 8 h, while those in group B (n=20) practiced using the chicken skin model for 8 h. The trainees' skills were assessed using the porcine model before and after training. Results Compared with the chicken skin model, this new training model more accurately resembled the structure and characteristic of human pelvis, urethral stump, and bladder (neobladder). After the training sessions, both groups improved in anastomosis time [GroupA: (64±11)min vs. (123±20)min, P＜0.05; Group B: (77±12)min vs. (121±17)min, P＜0.05] and quality (Group A: 8.8± 1.0 vs. 3.8 ± 1.2, P＜0. 05 ; Group B: 7.7 ± 0.9 vs.3. 7± 1.1, P＜0. 05). Compared with trainees in group B, trainees in group A required less time and achieved a higher quality score (P＜0.05). Conclusions This new training model can help urologic surgeons to reduce learning curve of this technique and improve their suturing skills. It is an effective,convenient training model for laparoscopic urethrovesical anastomosis.

Objective To obtain shRNA sequences that can stably block the expression of Nuclear Factor kappa- B (p65) in the prostate cancer cell line LNCaP and construct the lentivirus vector.And validate the gene function of p65 in the cell line. Methods According to p65 genetic information, we design siRNA1, siRNA2, siRNA3 those three siRNA sequences targeting the ods area of p65 gene and then form the corresponding four pairs of complementary single strand DNA of shRNA, including the sense strand and the antisense strand. The synthetic shRNA sequence was inserted into the empty pSIH1-H1-copGFP shRNA Vector, and after transfecting the prostate cancer cells , the inhibitory effect of p65 mRNA by different sequences was detected through real-time PCR, and the inhibitory effect of p65 protein expression was detected by Western-blotting. Thus we can obtain highly effective shRNA sequences in the inhibition of p65 in prostate cancer cells. MTT, flow cytometry, transwell were chosen to test the cell growth, migration and invasive power in vitro to compare the difference of the experimental group, control group and negative group. Results The third shRNA sequence had the best inhibitory effect and the inhibitory effect of p65 mRNA in prostate cancer cell line was 59 % and the protein was 81%. It's position locates in p65 (NM_021975 ) 1096-1113 and it's stemloop sequence is 5'-GATCCGCCCTATCCCTTTACGTCATTCAAGAGATGACGTAAAGGGATAGGGCTTTTTG-3'. After transfecting, the prostate cancer cell line had the low expression of p65 stably. Through MTT, we got the growth curve, which showed that the growth ability of experimental group was significantly decreased compared with the control group and the Logarithmic growth didn't appear in the first 96 hours. Flow cytometry test displayed that the percentage of G0-G1-phase cells in experimental group was 61.49%, and the control group was 44.89%, idle group was 41.52%, which was increasing oberviously. The S-phase cells in the experimental group was 28.58%, compared with the 47.36% and 46. 10% diminished. The results of transwell showed that the experimental group had 16. 5000±6. 62076 cells and the other two groups had 45. 6333 13. 54159 and 36. 8333±5. 68412 cells, which showed the invasive power of experimental group was significantly declined(P＜0.05).Conclusions It's successful to obtain shRNA sequences that can stably block the expression of p65 in the prostate cancer cell line LNCaP and construct the lentivirus vector. p65 can positively regulates the biological behavior of prostate cancer LNCaP cell line in the cell growth, migration and invasive power.

Objective To compare the efficacy,toxicitis and side-effects of Casodex and Flutamide in the hormonal therapy of advanced prostate cancer patients.Methods One hundred and thirty-six advanced prostate cancer patients were treated with with hormonal therapy.The patients were divided into 3 groups,of which 52 patients (group A) used LHRHa and Casodex as intermittent hormonal therapy;60 patients(group B) used LHRHa and Flutamide as intermittent hormonal therapy;24 patients(group C) were treated with surgical castration only.The difference of clinical symptoms,serum PSA,disease progression risk,survival rate,toxicitis and side-effects of 3 groups were compared.Results The relief rates of group A and B were 80.8% (42/52)and 81.7% (49/60) respectively,higher than 70.8% (17/24) of group C.The mean serum PSA of group A and B decreased from 133.3 ng/ml(17.9-982.8 ng/ml) to 15.8 ng/ml(0.02-28.9 ng/ml),142.6 ng/ml (20.2-1001.0 ng/ml)to 16.1 ng/ml(0.07-53.8 ng/ml),respectively,both better than that of group C,which decreased from 142.3 ng/ml (27.1-988.0 ng/ml) to 27.6 ng/ml(6.0-62.1 ng/ml).The mean chemical recurrence rates of group A and B were 34.7% (18/52) and 36.7% (22/60),respectively,lower than 58.3% (14/24) of group C.The mean chemical recurrence time of group A and B was 22(5-52)months and 22(6-65)months,respectively,longer than 11(5-54)months of group C.The mortality rates of group A and B were 26.9% (14/52) and 31.7% (19/60),respectively,lower than 66.7%(16/24) of group C.88.5% (46/52)of group A were treated continuously,while group B had 66.7% (40/60).The side-effects rate of group A was lower than group B.Conclusions Both Cadosex and Flutamide are effective for prostate cancer,and decrease the disease progression risk.Casodex is more effective and safer as for the treatment of prostate cancer compared to Flutamide.

Objective To detect the differentially expressed microRNAs (miRNAs) in bladder cancer tissue and normal bladder tissue. Methods Total RNA was extracted from bladder cancer tissue and normal bladder tissue by Trizol, and then miRNAs were isolated and enriched from the total RNA. Mammalian miRNA microarrays were used to analyze the differentially expressed miRNAs between the bladder cancer tissue and normal tissue. Data analysis was performed by software of LuxS-can3. 0 and SAM version 2. 1. Choose let-7 gene which was interesting to us, and validation of mi-croarray results was carried out by northern blotting. Results Compared with normal bladder tissues, there were 71 differentially expressed miRNAs in bladder cancer tissue, of which there were 38 down-regulated ones and 33 up-regulated ones. Among these miRNAs, 26 miRNAs were the most significant with 12 up-regulated and 14 down-regulated. The expression of let-7 gene in bladder cancer was down-regulated to normal bladder tissue by northern blotting, which was in agreement with the results of the miRNA microarrays. Conclusion There are differentially expressed miRNAs between bladder cancer tissue and normal bladder tissue, and let-7 gene is probably as a tumor suppressor in bladder cancer.