Pelvic exenteration for primary and recurrent malignancies in urology department / 中华泌尿外科杂志

Objective The objectives of present study is to investigate the safety and efficacy of pelvic exenteration (PE) for the treatment of pelvic malignancies in urology department.Methods From April 2010 to December 2014,20 patients with primary or recurrent pelvic malignancy accepted anterior pelvic exenteration (APE) or total pelvic exenteration (TPE) surgery,including 7 males and 13 females,ranged from 35 to 87 years old with an average of 65 years old.Ten case accepted APE and 10 for TPE.The ilium conduit was done in 5 cases for APE and 6 cases for TPE as urinary diversion,cutaneous ureterostomy was done in 5 cases for APE and 4 cases for TPE as urinary diversion.There were 6 cases primary tumor in APE group and 3 primary tumors in TPE.All of the patients had 13 cases of the urinary tract tumor group,and none of the urinary tract tumor group in 7 cases.There were 4 cases received preoperative chemotherapy in the urinary tract tumor group.No case received preoperative radiotherapy.3 cases received preoperative chemotherapy in none of the urinary tract tumor group,3 cased received preoperative radiotherapy.After induction of general anesthesia using a laryngeal mask for airway management.All patients took the abdominal incision,then dissected lymph nodes on both sides of the iliac vessels,freed bilateral ureters to the end of the swollen bladder,separated the peritoneal space.The bilateral vas deferens was cutted and ligated,then isolated and ligated the seminal vesicles between the posterior wall of the bladder and the anterior wall of the rectum.Lateral ligaments of bladder was cuted,then cuted ligament of prostate and puboprostatic ligament,sutured and cut deep vein of penis.Urethra of apex prostate was freed and cuted.Female patients needed to free the uterus and the posterior wall,cut the cardinal ligament and round ligament of uterus,isolate the posterior wall of the uterus to the posterior vaginal wall.Rectal resection adopted Miles operation.And sigmoid colostomy was performed on the left side of the abdominal wall.The perioperative characters,pathological results and patients' survival data were collected and analyzed.Results The average operation time for APE was 3.8 hours and 5.2 hours for TPE (P =0.173).Median length of hospital stay was 17.9 (7-47) days.The median blood loss was 300ml (80-2 500 ml) for APE and 400ml (50-6 000 ml) for TPE (P =0.909).The median follow-up time was 12.5 months (1-41months).The estimated 2-year survival rate for APE was 55.6％ and 45.0％ for TPE (P =0.642).Urinary system tumors group and non urinary system tumors group were analyzed and compared,The median survival time was 28 months and 13 months (P =0.538) in the two groups.The incidence of gastrointestinal complications of urinary system tumors and non-urinary system tumors was 7.7％ and 28.6％,incision complications was 7.7％ and 28.6％.Complications of urinary diversion only occurred in the non urologic tumor group,the incidence was 14.3％.The incidence of transfusion in two groups was 46.2％ and 28.6％.Conclusions Pelvic exenteration (APE and TPE) could be a safe and reliable choice for local advanced primary and recurrent pelvic malignancy.Even for the recurrent malignancies,the survival results of the patients were satisfactory.

Sunitinib induces autophagy via suppressing Akt/mTOR pathway in renal cell carci-noma / 北京大学学报(医学版)

Objective:To determine the mechanism of sunitinib-induced autophagy in renal cell carci-noma cells.Methods:MTS assay was applied to detect the cell viability alteration under the treatment of sunitinib (2,8 μmol /L).The sunitinib-induced autophagy as well as cell apoptosis was measured and compared after knocking down autophagy-related protein Beclin1 and microtubule associated protein 1 light chain 3 fusion protein (LC3)by RNA interference.The transmission electron microscope was used to observe the formation of autophagosomes in ACHN cells.The fluorescence microscope was used to mo-nitor distribution and aggregation of endogenous LC3-Ⅱ.The expressions of protein such as LC3-Ⅱ,the autophagic regulation molecules protein kinase B /mammalian target of rapamycin (Akt/mTOR)and the symbol of apoptosis poly ADP-ribose polymerase (PARP)were capable to be detected by immunoblotting assay.Results:Sunitinib was able to significantly trigger cell viability loss in the renal carcinoma cell ACHN,which was both in a concentration-dependent and time-dependent manner (P <0.05 ).After reducing the autophagy by knocking down Beclin1 and LC3,the number of cleavage of PARP was in-creased remarkably,whereas there was nearly not any cleavage in the mock group.By the transmission electron microscope,there were more autophagic vacuoles in ACHN cells after being administrated with sunitininb compared with the control.And the nuclear-to-cytosol translocation as well as aggregation of LC3-Ⅱ was presented after sunitinib treatment by the fluorescence microscope,which was the proof of the enhanced autophagy.According to the immunoblotting,sunitinib was able to increase the accumula-tion of LC3-Ⅱ.At the same time,the result of sunitinib combined with chloroquine,a drug which blocked the fusion of autophagosomes and lysosomes,demonstrated that the increasing amount of LC3-Ⅱwas due to the enhanced autophagy flux by sunitinib treatment in ACHN cells.However,phosphorylation of Akt as well as mTOR was decreased at the same time.The rapamycin (mTOR inhibitor)or knocking down Akt subunits could change the sunitinib-induced LC3-Ⅱ accumulation,whereas overexpression of Akt subunits decreased the autophagic flux,indicating that Akt/mTOR was the target of sunitinib in auto-phagy.Conclusion:Sunitinib induced autophagy via suppressing Akt/mTOR pathway,and the auto-phagy was involved in apopotosis.

Study on the Preparation Process of Zhenjing Xiehuo Granules / 中国药师

Objective: To investigate the preparation procedure of Zhenjing Xiehuo granules. Methods: Using the dry extract yielding rate and the contents of liquiritin and salvianolic acid B as the indices, an orthogonal test was adopted to choose the best ex-traction and purification technology. Using the qualified ratio of granules as the index, an orthogonal test was adopted to choose the best preparation process of the granules. Results:The optimized preparation conditions were as follows:Pulvis ferri was decocted first for 60 min. The other medicines were dipped in 8-fold amount of water for 90 min, and then added into pulvis ferri extracts and decocted for 3 times with 90 min for each time. The extracts were collected and concentrated till the relative density was 1. 3 (measured at 60℃), water was added with the dilution ratio of 1:2, ethanol was added till the percentage of ethanol was 50%, and then the mixed liquid was filtered after 24 hours. After ethanol was recycled from the filtrate, the filtrate was concentrated till the relative density was 1. 3 (measured at 60℃), and then dried at 60℃. Starch as the diluent, the ratio of extract to excipient was 1:0. 8, and the wet granules were prepared with 90% ethanol as the wetting agent, dried 3 hours at 60℃ followed by size stabilization to obtain the products. Con-clusion:The optimized preparation procedure of Zhenjing Xiehuo granules is stable and feasible.

Comparison of autophagy related proteins expression between clear cell renal cell carcinoma and adjacent tissue / 中华泌尿外科杂志

Objective To investigate the autophagy capacity in clear cell renal cell carcinoma tissue compared with adjacent normal tissue.Methods Sixty-nine pairs of samples from human clear cell renal cell carcinoma tissues and relatively healthy renal tissues adjacent to the tumor were collected during surgical resection.The expressions of proteins that were participating in the regulation and execution of autophagy were detected by immunohistochemisty.Electron-microscopy was also carried out for the morphometrical analysis.Results The protein expression of p-mTOR (P =0.004),P62 (P =0.000) and ULK1 (P =0.000) were up-regulated in the carcinoma tissue,while the expression of Beclin1 (P=0.000),LC3 (P =0.000) and ATG7 (P =0.000) were down-regulated,and the expression of ATG5 (P =0.349) had no signif-icant difference compared with adjacent normal tissue.Morphometrical analysis showed that the basal autophagic activity (measured by the presence of autophagy vacuole compartment) was remarkably down-regulated in carcinoma tissue,compared with adjacent normal tissue.The expression level of mTOR was correlated with P62,LC3 and ATG7,but results showed no correlation between mTOR and Beclin 1.Conclusion Our studies show a relatively reduced autophagy capacity in clear cell renal cell carcinoma,which is regulated by multiple autophagy-related proteins such as mTOR,Beclin 1 and LC3.

Distribution of positive lymph nodes of muscle invasive bladder cancer: analysis the pathological data from 208 consecutive cases of radical cystectomy / 中华泌尿外科杂志

Objective To describe the distribution of positive lymph nodes of muscle invasive bladder cancer, and explore the relationship between positive nodes and pathological characters. Methods Pathological data from 208 consecutive cases of muscle invasive bladder cancer were collect-ed and reviewed. The correlation of tumor grade, tumor stage and lymph nodes status was analyzed. The locations and numbers of positive nodes were recorded and compared according to the specific grade or stage. Results There were 153 cases (73.6%)of G_3 tumor and 55 cases(26.4%) of G_2 tumor and none G_1 (0%)in this cohort. The case number from pT1 to pT4 was 59(28. 4%)、58 (27.8%)、48(23.0%)and 43(20.6%), respectively. The tumor grade was positively correlated with tumor stage in this cohort (r=0. 392, P=0. 000). 153 cases had been taken lymph node dissection. There was more node positive cases in pT_3 and pT_4 than that in T_1 (P=0. 001 ,P=0. 000), as well as pT_4 compared with pT_2 (P= 0. 012). The data showed that most of the positive nodes were located within the pelvic region. There was only 1 case and 1 node positive for G_1/G_2 tumor with 24.84% of node positive cases for G_3. The positive nodes involved from pelvic to proximal artery while the stage increased. Conclusions There is less chance for low grade (G_1/G_2) bladder cancer to be node posi-tive compared with G_3 ones. It is necessary to take a extensive lymphadenectomy for the patients with stage more than T_2.

Anatomical retroperitoneoscopic nephrectomy——with clinical experience of 405 cases / 中华泌尿外科杂志

Objective To evaluate the safety and efficacy of the anatomical retroperitoneoscopic nephrectomy(RSN)and standardize the procedure of RSN. Methods The retrospective analysis was performed on 405 consecutive patients underwent anatomical RSN in Our institute from January 2002 to June 2008.There were 232 male and 173 female patients with the average age of(57.2±14.2)years,among whom there were 228 renal cell carcinoma patients accepted RSU,96 and 49 renal pelvic carcinoma and ureteral carcinoma cases accepted retroperitoneoscopic ureteronephrectomy (RSUN) and 32 cases accepted simple RSN due to loss of renal function caused by benign renal discsses.The tadical RSN was performed by dissecting outside Gerota's fascia and in the latent cavities between this fascia and lateral conal fascia in the dorsal side and between this fascia and prerenal fusion fascia in the ventral side,whereas the simple RSN was done inside Gerota's fascia by making direct incision on it and dissecting between this fascia and perirenal adipose tissue.Kidneys and perirenal adipose tissue were completely removed by dissection along several avascular planes around the kidney under the amplified view of laparoscopy. The software SPSS 12.0 was used for the statistical analysis of all data. Results The mean operative time was (132±48)min for radical and simple RSN and (245 ± 62)min for radical RSUN, which included the time for position change and second skin preparation. The medium estimated blood loss was 100 ml(10-2500 ml) and the average drainage volume was 150 ml (0-1152 ml) postoperatively. 15 cases (3. 70%) required blood transfusion with the median volume of 400ml (400-1650 ml). Four cases (0. 99%) were converted to open surgery due to severe adhesion (2 cases), difficult exposure of renal helium (1 case) and severe bleeding (1 case).The mean drainage time was (3. 9±1.8)d, the mean time to first oral intake was (2.7±1.2)d and the mean postoperative hospital stay was (8.6±3. 8)d. Conclusion The anatomical RSN is safe and effective and should be the standard surgical procedure for laparoscopic nephrectomy.

Inhibition of autophagy leads to increased apoptosis in LNCaP cells under androgen deprivation conditions / 中华泌尿外科杂志

Objective To investigate the change of apoptosis in LNCaP cells after inhibition of autophagic process under androgen removal conditions. Methods The autophagic level was deter-mined by using confocal microscopy and RT-PCR. The DAPI staining was used to indicate the apopto-sis of LNCaP cells after inhibition of autophagic by 3-MA. Also, Z-VAD-FMK was used to extend the apoptosis results. Results ①Androgen deprivation led to increased autophagy in LNCaP cells. LN-CaP cells cultured in complete medium(CM) presented low autophagic process with 1.9 scores. After 24 hours, the punetate GFP-LC3 structures were accumulated in the cells cultured in serum-free medi-um (SF)(2.64 scores). In contrast, the number of punctate GFP-LC3 remained at a very low level (1.85 scores), when cells were incubated with DHT in SFA(serum-free medium+DHT). Statistical analysis showed the significant difference between SF and SFA (P<0.01). Semiquantitative RT-PCR was employed to examine the mRNA expression of LC3. Indeed, cells grown in the medium without serum had a higher LC3 mRNA expression with the highest at 12 hour time point as compared with the cells grown in CM. DHT treatment reduced the level of LC3 mRNA. ②Blockage of autophagy by 3-MA increased the apoptosis of LNCaP cells. LNCaP cells in SF and SFA just presented a basal level of apoptosis, which is (3.19±1.09)% and (3.01±0.33)% , respectively. Under androgen-free con-ditions, inhibition of autophagy by 3-MA could increase apoptosis significantly(10. 90±2.91%). While Z-VAD-FMK, a pan Caspase inhibitor, was able to suppress this apoptotic process to the level of (1.16±0.52)%, which was statistically significant(P<0.01). Conclusions Androgen removal can lead to the increase of autophagy in LNCaP cells. Moreover, inhibition of autophagy promotes the occurrence of apoptosis.

Renal cell carcinoma related novel gene, GYLZ-RCC18: cloning and functional studies / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To clone the full length of renal cell carcinoma (RCC) related novel gene GYLZ-RCC18 and study its function.</p><p><b>METHODS</b>SMART RACE technology was used to clone the full length of GYLZ-RCC18. RT-PCR was used to detect its expression in renal cell carcinoma tissue at different stages and grades. We transfected the antisense oligonucleotide of GYLZ-RCC18 to renal cell carcinoma cell line, GRC-1, and analyzed proliferation activity, growth rate, apoptosis, and mortality changes.</p><p><b>RESULTS</b>The full length of GYLZ-RCC18 (GenBank accession number: BE825133) cDNA was about 3.5 kb. GYLZ-RCC18 had a higher expression in higher grades and stages of renal cell carcinoma than in lower ones. The expression of GYLZ-RCC18 in renal cell carcinoma was much higher than in normal kidney. After the transfection of GYLZ-RCC18 antisense oligonucleotide, the mortality of GRC-1 increased significantly, while proliferative activity and growth rate were substantially inhibited at the same time. The antisense oligonucleotide induced apoptosis of GRC-1 through the entire observation time.</p><p><b>CONCLUSION</b>GYLZ-RCC18 is an important novel gene related to renal cell carcinoma. Overexpression of this gene results in higher growth and proliferative activity and has an antiapoptosis effect on renal cell carcinoma cells. Transfection of the antisense oligonucleotide may inhibit the generation and development of renal cell carcinoma.</p>