Purpose　The aim is to study the activity and the sildenafil selective inhibition of PDE5. Methods　The PDE isoenzymes were purified from bovine penis corpus cavernosum tissue by FPLC system. PDE activity was assayed by using 3　H-cGMP as substrate, the PDE isoenzymes hydrolyzed it to 3　H-GMP, and 3　H-GMP was further hydrolyzed to 3　H-guanosine by 5′-nuclease of snake venom. Add scintillation cocktail to observe the PDE isoenzymes activity. The selective inhibitor sildenafil of different concentrations were used to observe the inhibition of PDE5. Data replotted according to procedure of Dixon plots.Results　Three PDE isoenzyme peaks were purificated from bovine corpus cavernosum. The PDE of the third peak had the strongest activity of cGMP hydrolyzation which could be inhibited by sildenafil apparently.Conclusion　Since PDE5 was mainly found in corpus cavernosum tissue of mammalian, and sildenafil was a selective inhibitor of PDE5. It was suggested that the third peak was PDE5. The result was in agreement with the article reported.

Vascular endothelial cells (EC) and smooth muscle cells (SMC) are target and effecter cells of inflammation, and they play an important role in inflammatory responses. The abnormal structure and function of EC and SMC play a significant role in microcirculation disturbance in septic shock and multiple organ dysfunction. This review was meant to discuss the changes in structure and function of EC and SMC and their bidirectional regulation. The cellular linkage of EC and SMC is essential for the interactions between them, and it contributes to the course of sepsis. Paracrine and autocrine as produced by EC and SMC constitute a network for mutual adjustment. Replication of the interaction between EC and SMC facilitates the potential to support hemodynamics, tissue perfusion and cellular metabolism, thereby lower the mortality rate of sepsis. However, the detailed and specific mechanisms remain to be disclosed.

Objective To compare the perioperative outcomes and short-term efficacy of three-port extraperitoneal laparoscopic radical prostatectomy (ELRP) and four-port ELRP.Methods Two hundred patients who had undergone ELRP for prostate cancer by a single surgeon from November 2010 to October 2014 were retrospectively analyzed.Among them,95 cases underwent three-port ELRP and 105 cases underwent four-port ELRP.On the basis of traditional four-port ELRP,three-port ELRP was characterized by the omission of the trocar on the inner side of right anterior superior iliac spine.The mean age was 66.8 ± 15.5 years,and mean total prostate specific antigen (tPSA) was 15.3 ± 12.4 μg/L.There were no significant differences including age,body mass index,tPSA,clinical stages,acceptance of neoadjuvant hormone therapy,history of transurethral resection of the prostate,history of diabetes mellitus between the 2 groups (P ＞ 0.05).Patients in three-port ELRP group had significantly smaller prostate volume than fourport group (35.6 ± 16.7 ml versus 42.2 ± 24.7 ml,P ＜ 0.05).The clinical factors as operative time,estimated blood loss,hospital stay,drainage tube keeping days,pathological Gleason scores,pathological stages,positive surgical margin rates,biochemical recurrence rates and urinary incontinence rates were compared between the 2 groups.Results The three-port group had significantly shorter operative time than the four-port group (81.0 ± 18.6 min versus 103.6 ±34.6 min),less estimated blood loss (102.6 ±75.8 ml versus 217.5 ± 182.9 ml),less positive surgical margin rates (13.7％ versus 27.6％).There were 9 patients having Gleason scores more than 7 in the three-port ELRP group and 29 patients in four-port ELRP group (P ＜ 0.05).There were no significant differences of hospital stay,drainage tube keeping days,pathological stages between the 2 groups (P ＞ 0.05).Eighty-three cases in the three-port ELRP group (87.4％) were followed up for 5-19 months with the median time of 11 months.Ninety-two cases in fourport ELRP group (87.6％) were followed up for 17-52 months and the median time was 27 months.There were no significant differences of biochemical recurrence rates and urinary incontinence rates between the 2 groups(P ＞ 0.05).Conclusions Compared to four-port ELRP,three-port ELRP can provide shorter operative time,less blood loss,better negative surgical margin rates,similar oncological control and recovery of postoperative continence.In experienced hands,three-port ELRP could be a feasible and effective option for localized prostate cancer.

Objective To evaluate the learning curve of three-port extraperitoneal laparoscopic radical prostatectomy(ELRP) and to minimize operative time and blood loss about this procedure.Methods From August 2013 to October 2014,the data from 95 consecutive patients,who had undergone three-port ELRP for prostate cancer,were retrospectively analyzed.The mean age was 65.9 ± 7.7 years,mean total PSA level was 15.4 ± 12.7 μg/L,and mean body mass index(BMI) was 24.8 ± 3.2 kg/m2.According to the number of procedures performed by the surgeon,all patients were classified into three chronologic groups,including group A (No.1-32),group B (No.33-64) and group C (No.65-95).There were no significant differences including age,BMI,tPSA,estimated prostate volume,clinical stages,history of neoadjuvant endocrine therapy,history of transurethral resection of the prostate (TURP) among group A,B and C (P ＞ 0.05).The operative outcomes analyzed were operative time,estimated blood loss,hospital stay,drainage tube indwelling days,pathological Gleason scores,pathological stages,positive surgical margin rates,biochemical recurrence rates and urinary incontinence rates.Among these 95 patients,the results of the first 32 cases were compared with those of the remaining 63 cases,the first 64 with the remaining 31.Results The average operative time in 95 patients was 81.0 ± 18.6 min.The sloping learning curve for this surgeon showed that the operative time for all 95 cases was strongly correlated with additional experience (| rs | =0.612,P＜0.01).Operative time,however,was not strongly correlated with the surgeon's experience in each group of A,B and C(P ＞0.05).Group A had longer operative time than that of Group B plus C(96.4 ± 11.3 min vs 73.2 ± 16.7 min,P ＜0.01).Group A plus B had longer operative time than that of group C (87.6 ± 17.2 min vs 67.5 ± 13.8 min,P ＜ 0.01).For all cases,the estimated blood loss was strongly correlated with additional experience (| rs | =0.677,P ＜ 0.01).Estimated blood loss was strongly correlated with the accumulation of experience for the initial 32 cases(| rs | =0.619,P ＜ 0.01).However,no strong correlation was observed over the next 63 cases.Group A had more blood loss than that of Group B plus C (158.7 ± 81.3 ml vs 74.1 ± 54.4 ml,P ＜ 0.01).Group A plus B had more blood loss than that of group C (125.5 ± 71.6 ml vs 55.3 ± 61.6 ml,P ＜ 0.01).But hospital stay,drainage tube keeping days were not strongly correlated with additional experience in each group(P ＞ 0.05).There were no significant correlation between the accumulation of experience and positive surgical margin rates,biochemical recurrence rates and urinary incontinence rates.Conclusion Our experience of three-port ELRP cases appears to be favorable with decreasing tendency in operative time,estimated blood loss with experience accumulation.Exposure to 32 surgeries,operative time and estimated blood loss reduced significantly,and after 64 cases operative time and estimated blood loss further reduced.

Objective To investigate the effects and mechanisms of omega-3 polyunsaturated fatty acids (ω-3 PUFA) supplementation on colonic macroscopic and histological score , inflammatory response , and endo-plasmic reticulum stress ( ERS ) response in experimental rat models of colitis .Methods Experimental rat models of colitis were induced by trinitro-benzene-sulfonic acid (TNBS).Totally 100 male SD rats were ran-domly divided into 5 groups according to the random data tables:sham operation group ( Sham group ) , inflam-matory bowel disease group (IBD group),ω-3 PUFA supplementation group (IBD+ω-3 group), 5-aminosali-cylic acid group ( IBD +5-ASA group ) , and ERS activation 2-deoxy-D-glucose group ( IBD +ω-3 +2-DG group).Colonic macroscopic and histological scores were evaluated on days 1, 3, 7 and 14 after modeling.The serum levels of tumor necrosis factor-α(TNF-α), interleukin (IL) -1, and IL-6 were measured using en-zyme-linked immunosorbent assay , whereas ERS cytokines including glucose-regulated protein 78 ( GRP78 ) , inositol-requiring enzyme 1 (IRE-1), and C/EBP homologous protein (CHOP) were tested by Western blot. Results Compared with the Sham group , colonic macroscopic and histological score , the serum levels of in-flammation relatived factors (TNF-α, IL-1, IL-6) and ERS relatived factors (GRP78、 IRE-1, CHOP) were significantly increased on the rest of the four groups ( all P<0.001 ) .Compared with the IBD group , ω-3 PUFA supplementation reduced colonic macroscopic [7 d: 3.55 ±0.29 vs.4.37 ±0.39, P=0.03, 14 d:2.46 ±0.17 vs.3.86 ±0.21, P=0.04] and histological score [7 d: (2.56 ±0.27) scores vs.(3.45 ± 0.40) scores, P=0.02, 14 d: (2.23 ±0.20) scores vs.(3.06 ±0.26) scores, P=0.04].Meanwhile,ω-3 PUFA supplementation suppressed the expressions of inflammation [TNF-α:(43.71 ±11.39) pg/ml vs. (84.97 ±13.81) pg/ml, P=0.02, IL-1:(38.51 ±10.60) pg/ml vs.(73.04 ±12.48) pg/ml, P=0.01, IL-6:(28.91 ±7.27) pg/ml vs.(53.45 ±9.40) pg/ml, P=0.02] and ERS relatived factors (GRP78:2.41 ±0.29 vs.1.47 ±0.21, P=0.01, IRE-1:2.83 ±0.31 vs.1.23 ±0.20, P<0.001, CHOP:1.89 ± 0.17 vs.1.32 ±0.11 , P=0.04 ) .However , the salutary effects of ω-3 PUFA would been reversed by ERS activation 2-deoxy-D-glucose [ TNF-α: (72.67 ±10.37 ) pg/ml vs.(43.71 ±11.39 ) pg/ml, P =0.02, IL-1:(57.66 ±13.88) pg/ml vs.(38.51 ±10.60) pg/ml, P=0.02, IL-6: (46.10 ±9.67) pg/ml vs. (28.91 ±7.27) pg/ml, P=0.01, GRP78:1.47 ±0.21 vs.1.82 ±0.24, P=0.03, IRE-1:1.23 ±0.20 vs.2.21 ±0.23, P=0.02, CHOP:1.32 ±0.11 vs.1.61 ±0.16, P=0.04].Conclusion The salutary effects of ω-3 PUFA supplementation on the colitis induced by TNBS appear to be mediated by inhibited inflam -matory responses , which may suppress the activation of ERS response .

Objective:To explore an improved osteotomy of mandible ascending ramus for the surgical approach to large tumors in parapharyngeal space.Methods:According to the operation of parotid gland, masseter muscle was cut near the border of mandibular angle till subperiosteum dissection to mandibular notch, vertical osteotomy outside mandibular foramen to 1.0 cm under mandibular notch, vertical to posterior border of ascending ramus, tumor was exposed and removed, bone plate was repositioned and fixed with titanium. Results:Tumors were completely removed in this way in 3 patients without complications. Conclusion:This surgical approach is suitable for the surgical removal of parapharyngeal interstitial giantism tumor.

Objective To investigate the shape and hemodynamic characteristics of the ruptured posterior communicating artery minimal aneurysms. Methods The clinical data of 42 patients with ruptured posterior communicating artery minimal aneurysm (the maximum diameter < 3 mm,11 ruptured aneurysms and 31 unruptured aneurysms)were collected retrospectively. Three-dimensional DSA shapes of the aneurysms were assessed,and the hemodynamic parameters of the aneurysms were calculated according to their computer simulation models. Results (1)The multiple aneurysms were more common in the unruptured group than those in the ruptured group (58. 1% [18/ 31]vs. 9. 1% [1/ 11]). There was significant difference,P = 0. 006 ). (2 )The complex flow pattern was more common in the ruptured group (63. 6%[7 / 11]vs. 6. 5% [2 / 31],P < 0. 01)and also the changed flow pattern (45. 5% [5 / 11]vs. 3. 2% [1 / 31),P = 0. 003). (3)The median aneurysm wall shear stress of the ruptured group was 0. 74 (0. 52,0. 86)and that of the unruptured group was 1. 03(0. 83,3. 64). There was significant difference between the 2 groups (P =0. 008). Conclusion The unruptured minimal aneurysms are common in patients with multiple aneurysms. Active surgical intervention is recommended for the posterior communicating artery minimal aneurysms with low wall shear stress,complex and change flow.

Objective To compare the perioperative outcomes and postoperative complications of retroperitoneal laparoscopic partial nephrectomy ( RLPN) for the treatment of endophytic renal tumors and non-endophytic tumors.Methods Three hundred and ninety-two patients who underwent RLPN for kidney neoplasms from May 2005 to September 2012 were retrospectively analyzed . They were divided into endophytic renal tumor group ( 48 cases ) and non-endophytic tumor group ( 344 cases ) .There were no significant differences in the aspects of gender , age, body mass index, tumor side, diameter, preoperative estimated glomerular filtration rate (eGFR) between the two groups.Operative time, warm ischemia time, method of renal vascular occlusion , repair rate of renal collecting system , estimated blood loss , usage of laparoscopic ultrasonography , intraoperative complications , pathological types , postoperative hospitalization days, postoperative complications and postoperative eGFR were collected and analyzed .Results Patients with endophytic tumors had significantly more usage of laparoscopic ultrasonography (95.8%versus 1.2%, P<0.001) and higher repair rate of renal collecting system (35.4%versus 6.1%, P<0.001).Clamping segmental renal artery and without clamping renal vessels were not used in dealing with renal vessels of endophytic tumors (P<0.05).There were no significant differences of operative time , warm ischemia time, estimated blood loss , intraoperative complications , pathological types , postoperative hospitalization and postoperative eGFR (P>0.05) between the 2 groups.All the patients′surgical margins were negative.The rates of postoperative complication ( Clavien gradeⅠtoⅢb) were 4.2%and 2.9%in the endophytic group and non-endophytic group , respectively ( P >0.05 ) . Median follow-up was 42 ( 33 -108 ) months in endophytic group and 45 (33 -120) months in non-endophytic group.No local recurrence or metastasis occurred in the two groups .Conclusions In experienced hands , RLPN could represent a feasible , safe and effective treatment for selected patients diagnosed with endophytic renal tumors .Laparoscopic ultrasonography is valuable on locating the tumor and defining tumor margins in RLPN of endophytic renal tumors .

OBJECTIVETo explore the effects of exogenous carbon monoxide-releasing molecule 2 (CORM-2) on formation of human neutrophil extracellular traps (NETs) stimulated by endotoxin/lipopolysaccharide (LPS) and its relevant mechanism.

METHODSVenous blood samples were collected from a healthy adult volunteer to isolate neutrophils. The neutrophils were divided into normal control (NC) group, LPS group, LPS+ 10 μmol/L CORM-2 group, LPS+ 50 μmol/L CORM-2 group, and LPS+ inactive CORM-2 (iCORM-2) group according to the random number table. No treatment was given to the neutrophils in NC group. The neutrophils in LPS group underwent LPS stimulation (1 μL, 1 μg/mL). The neutrophils in LPS+ 10 μmol/L CORM-2 group, LPS+ 50 μmol/L CORM-2 group, and LPS+ iCORM-2 group underwent the same LPS stimulation as that in LPS group and treatment of 10 μmol/L CORM-2, 50 μmol/L CORM-2, and 50 μmol/L iCORM-2, respectively, with the volune of 1 μL. After conventional culture for 1 h, the number of NETs was determined with propidium iodide staining method; the early cell apoptosis rate was determined with flow cytometer; the generation level of reactive oxygen species (ROS) was assessed with dihydrogenrhodamine 123 fluorescent probe staining method (denoted as mean fluorescence intensity); the expression level of phosphorylated extracellular regulated kinase 1/2 (p-ERK1/2) was determined by Western blotting. The sample numbers of each group in the 4 experiments were all 5. Data were processed with one-way analysis of variance and SNK test.

RESULTS(1) The numbers of NETs per 400-time visual field in cells of LPS and LPS+ iCORM-2 groups were close to the number in NC group (with P values above 0.05). The number of NETs per 400-time visual field was significantly larger in cells of LPS+ 10 μmol/L CORM-2 and LPS+ 50 μmol/L CORM-2 groups than in NC and LPS groups (with P values below 0.05). The number of NETs per 400-time visual field in cells of LPS+ iCORM-2 group was close to that of LPS group (P>0.05). (2) The early cell apoptosis rate was significantly increased in LPS, LPS+ 10 μmol/L CORM-2, LPS+ 50 μmol/L CORM-2, and LPS+ iCORM-2 groups than in NC group (with P values below 0.05). The early cell apoptosis rates in LPS+ 10 μmol/L CORM-2, LPS+ 50 μmol/L CORM-2, and LPS+ iCORM-2 groups were close to the rate in LPS group (with P values above 0.05). (3) The generation level of ROS was significantly higher in cells of LPS, LPS+ 10 μmol/L CORM-2, and LPS+ iCORM-2 groups than in NC group (with P values below 0.05). The generation level of ROS in cells of LPS+ 50 μmol/L CORM-2 group was close to that of NC group (P>0.05). The generation level of ROS was lower in cells of LPS+ 10 μmol/L CORM-2 and LPS+ 50 μmol/L CORM-2 groups than in LPS group (with P values below 0.05), while the generation level of ROS in cells of LPS+ iCORM-2 group was close to that of LPS group (P>0.05). (4) The expression levels of p-ERK1/2 in cells of LPS and LPS+ iCORM-2 groups (respectively 0.0311±0.001 and 0.0309±0.0018) were close to the level in NC group (0.0304±0.0046, with P values above 0.05). The expression level of p-ERK1/2 was significantly higher in cells of LPS+ 10 μmol/L CORM-2 and LPS+ 50 μmol/L CORM-2 groups (respectively 0.7891±0.0201 and 1.2970±0.0056) than in NC group (with P values below 0.05). The expression level of p-ERK1/2 was significantly higher in cells of LPS+ 10 μmol/L CORM-2 and LPS+ 50 μmol/L CORM-2 groups than in LPS group (with P values below 0.05). The expression level of p-ERK1/2 in cells of LPS+ iCORM-2 group was close to that of LPS group (P>0.05).

CONCLUSIONSCORM-2 can obviously increase the production of NETs in LPS-induced neutrophils, and it might be attributable to the promotion of inhibition of ROS generation and phosphorylation of ERK1/2.

Apoptosis ; Carbon Monoxide ; metabolism ; Extracellular Traps ; Humans ; Lipopolysaccharides ; pharmacology ; Organometallic Compounds ; pharmacology ; Phosphorylation ; drug effects

Objective:To analyze the impact of different surgical methods on perioperative complication rate after radical cystectomy in patients over 75 years of age.Methods:A retrospective study reviewed 1 432 patients who received radical cystectomy from January 2003 to January 2015.A total of 239 pa-tients were ≥75 years (median age:78 years),of whom,74 patients (31.0%)suffered one or more perioperative complications.According to the different operation methods,patients could be divided into ileal conduit group and cutaneous ureterostomy group.The ileal conduit group included laparoscopic and open radical cystectomy with ileal conduit.The cutaneous ureterostomy group included transperitoneal laparoscopic,transperitoneal and extraperitoneal open radical cystectomy with cutaneous ureterostomy. Results:Perioperative complications were significantly associated with the patient’s age (P =0.012), American Society of Anesthesiologists (ASA)score (P =0.001),node staging (P =0.043),and dif-ferent surgical methods.Perioperative complications caused a prolonged hospital stay and delayed recovery (34 d vs.21 d,P =0.002).For different surgical methods,the perioperative complication rate of ileal conduit was higher than cutaneous ureterostomy (P =0.013).However,there were no significant differences between transperitoneal laparoscopic and open radical cystectomy with ileal conduit in perioperative complication rate (P =0.836).The perioperative complication rate was statistically signifi-cant among transperitoneal laparoscopic,transperitoneal and extraperitoneal open radical cystectomy with cutaneous ureterostomy (P =0.022).On multivariate analysis,age (hazard ratio =4.856,95% CI:1.465 -16.103,P =0.010),ASA score (P =0.008),and different surgical methods (P =0.016) were significantly associated with the perioperative complication rate.Conclusion:The perioperative complication rate after radical cystectomy in elderly patients was associated with the patients’age,ASA score,and different surgical methods.Patients who received extraperitoneal open radical cystectomy with cutaneous ureterostomy suffered fewer perioperative complications,which was an appropriate choice for elderly patients.