ObjectiveTo assess the feasibility of seeding adipose-derived stem cells (ADSCs) onto bladder acellular matrix grafts (BAMGs) for bladder reconstruction in a rabbit model.MethodsAutologous ADSCs were isolated,expanded and identified by flow cytometry.In the experimental group,ADSCs were seeded onto BAMGS for reconstructing bladder defects in 12 male rabbits.Unseeded BAMGs were used for bladder reconstruction in the control group of 12 rabbits.Cystography was performed at 24 weeks after grafts implantation.Following cystography,the animals were scarified and grafts were harvested； H&E and immunohistochemical staining were performed with cytokeratin AE1/AE3,smooth muscle α-actin and S-100 markers.ResultsFlow cytometry demonstrated that the ADSCs expressed CD90,CD44,CD105,CD166 and CD34,but not CD45 or CD106.The cells demonstrated good biocompatibility with BAMGs.At 24 weeks,in the experimental group,the reconstructed bladders reached a mean volume of (94.68 ± 3.31 )％ of the precystectomy bladder capacity.Complete regeneration of smooth muscle and nerve tissue was evident.Regenerated SMCs,urothelium and nerve cells stained positively for α-smooth muscle actin,AE1/AE3 and S100.In the control group,the mean bladder volume was (69.33 ± 5.05 )％ of the pre-cystectomy volume.Histologically,the control group was characterized by multi-layered urothelium without evidence for organized muscle or nerve tissue.Conclusion The tissue engineering bladder constructed by ADSCs and BAMG can be used as an ideal biomaterial to replace and repair the bladder.

OBJECTIVETo screen the lentiviral vector carrying siRNA and capable of significantly suppressing the ROCK2 gene expression in the corpus cavernosum smooth muscle cells of spontaneously hypertensive rats (SHR).

METHODSWe designed and synthesized 4 siRNA fragments targeting the ROCK2 gene and packaged them into lentiviral vectors. We collected corpus cavernosum smooth muscle cell samples from 5 male SHRs and randomly divided them into groups A (non-transfection control), B (GFP lentiviral transfection), C, D, E, and F (lentiviral transfection with siRNA fragments 1 -4 targeting the ROCK2 gene). Each group consisted of 5 samples and each sample 3 x 10(4) cells. At 48 hours after transfecting MOI = 80 into the SHR corpus cavernosum smooth muscle cells, we detected the expression of GFP under the fluorescent microscope and the mRNA expression of the ROCK2 gene by RT-PCR.

RESULTSThe transfection efficiency of the SHR corpus cavernosum smooth muscle cells was > 50%. Compared with group A, the expression of ROCK2 mRNA in the corpus cavernosum smooth muscle cells showed no remarkable change in group B (P > 0.05) but was inhibited very significantly in C ([43.91 +/- 8.19]%), D ([47.15 +/- 6.64]%), and F ([25.7 +/- 6.03]%) (P < 0.01), and significantly in E ([16.81 +/- 5.94]%, P < 0.05).

CONCLUSIONWe successfully constructed 4 lentiviral vectors carrying siRNA targeting the ROCK2 gene, all of which can significantly suppress the ROCK2 expression in the SHR corpus cavernosum smooth muscle cells, and one has a highly strong inhibitory effect.

Animals ; Gene Silencing ; Genetic Vectors ; Lentivirus ; genetics ; Male ; Myocytes, Smooth Muscle ; Penis ; cytology ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; Random Allocation ; Rats ; Rats, Inbred SHR ; Transfection ; methods ; rho-Associated Kinases ; genetics

OBJECTIVE:To compare the quality of domperidone tablets produced by 3 different factories METHODS:The quality of different products was investigated according to the standards of Ministry of Public Health and manufacture factories,including dissolubility,uniformity of dosage units,related substances and seals of blister packaging RESULTS:In 3 batches of products manufactured by Y,W and L,the contents of domperidone were 98 23%～99 74%,96 92%～98 31% and 97 42%～98 72%,and content uniformity A+1 8S was 2 11～3 38,3 97～5 25 and 5 49～7 47 respectively,which were in keeping with the Chinese pharmacopoeia;impurity R061668 was not found,and contents of impurity R052211 were 0 004%～0 024%,0 029%～0 072% and 0 003%～0 056%;the total impurity substances were 0 030%～0 095%,0 102%～0 128% and 0 232%～0 489%;the dissolubity(45min) was 95 09%～95 77%,91 25%～96 43% and 95 48%～96 58% respectively In experimental condition,no leakage was found from blister package of Y and W products,however,the leak rate of L product reached more than 38% CONCLUSION:According to this survey,the quality of product Y is the best,W is better

Objective To evaluate short-term fowllow-up results of castable ceramic-IPS Empress Ⅱ onlays in restoring severe tooth defects with low occlusogingival dimension from clinical aspects.Methods 41 units of the onlay were fabricated and applied in 37 patients,who were divided into experimental group and control group.The experimental group were restored with IPS EmpressⅡcastable ceramic onlay which was bonded by a dual-cure adhesive system.The control group were restored with mental onlay which was bonded by glass-ionomer cements.All these cases were followed at 6 and 12 months after cementation of the restorations.Metewands were established to evaluate its clinical effects.Results Onlay made from IPS Empress Ⅱ castable ceramic demonstrated natural and esthetical appearance.In addition,proximal contact,retention and marginal fit of the restorations were good.No gingivitis was found.Conclusion The reliability of onlay made from IPS Empress Ⅱ castable ceramic is greatly improved due to the strengthening effects of the lithium disilicate structures.In addition,the better esthetics compared to resin filling technique makes it a comparatively ideal all-ceramic system for onlays.

OBJECTIVETo explore the relationship of bone cement distribution and the puncture angle in the treatment of thoracolumbar compression fractures with unilateral percutaneous kyphoplasty (PKP).

METHODSThe clinical data of 37 patients with thoracolumbar osteoporotic compression fractures underwent PKP between January 2013 to March 2014 were retrospectively analyzed, all punctures were performed unilaterally. There were 6 males, aged from 65 to 78 years old with an average of (71.83 ± 6.15) years; and 31 females, aged from 57 to 89 years old with an average of (71.06 ± 7.89) years. Imaging data were analyzed and puncture angle and puncture point were measured before operation. According to the measured data, the puncture were performeds during the operation. Distribution area of bone cement were calculated by X-rays data after operation. The effect of bone cement distribution on suitable puncture angle was analyzed; VAS score was used to evaluate the clinical effects.

RESULTSThe puncture angle of thoracic vertebrae in T8-T12 was from 28° to 33° with an average 30.4°; and the puncture angle of lumbar vertebrae in L1-L5 was from 28° to 35° with an average of 31.3°. Postoperative X-rays showed the area ratios of bilateral bone cement was 0.97 ± 0.15. Bilateral diffuse area were basic equal. Postoperative VAS score decreased significantly (1.89 ± 1.29 vs 7.03 ± 1.42).

CONCLUSIONThrough measure imaging data before operation with PKP,the puncture point and entry point can be confirmed. According the measured data to puncture during operation, unilateral puncture can reach the distribution effect of the bilateral puncture in the treatment of thoracolumbar compression fractures.

Aged ; Aged, 80 and over ; Bone Cements ; Female ; Fractures, Compression ; surgery ; Humans ; Kyphoplasty ; methods ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Middle Aged ; Spinal Fractures ; surgery ; Spinal Puncture ; methods ; Thoracic Vertebrae ; injuries ; surgery

OBJECTIVETo observe the clinical effect of electric stimulation of long-term retaining needle on analgesia after laparoscopic cholecystectomy (LC) and the impacts on the post-surgical flatus time.

METHODSUnder static absorptive composite general anesthesia, 90 cases of LC were randomized into three groups, 30 cases in each one. In the control group, the analgesia was not applied after LC. In the analgesia-pumper group, the patient controlled intravenous analgesia (PCIA) was used. In the needle-retaining group, the electric acupuncture stimulator was used. The needles were inserted transversely at Riyue (GB 24), Qichong (ST 30) and Yanglingquan (GB 34) and fixed with sterile sticker. Separately, in 8 h and 24 h after surgery, the electric acupuncture stimulation with disperse-dense wave, 2 Hz/100 Hz frequency was applied continuously for 30 min. Visual analogue scale (VAS), adverse reactions such as vomiting and nausea and the postoperative flatus time in 2, 4, 8, 12, 24 and 36 h after surgery were observed and recorded in the three groups.

RESULTSIn 2, 4, 8, 12 and 24 h after surgery, VAS scores in the needle-retaining group and the analgesia-pumper group were all lower than those in the control group (P < 0.05, P < 0.01). The analgesia effect at the above time points in the needle-retaining group was better than that in the analgesia-pumper group (all P < 0.05). There was not adverse reaction in the needle-retaining group. But there were 3 cases of somnolence, 6 cases of nausea and 3 cases of vomiting in the analgesia-pumper group, and 2 cases of nausea and 1 case of vomiting in the control group. The flatus time was quite earlier in the needle-retaining group as compared with the other two groups [(14.77 +/- 4.99) h vs (18.50 +/- 4.22) h, P < 0.01; (14.77 +/- 4.99) h vs (18.17 +/- 4.69) h, P < 0.05].

CONCLUSIONThe electric stimulation of long-term retaining needle is safe and effective in analgesia after LC. It avoids the adverse reactions of analgesics and promotes postoperative flatus.

Acupuncture Analgesia ; instrumentation ; Adult ; Aged ; Cholecystectomy, Laparoscopic ; adverse effects ; Electroacupuncture ; instrumentation ; Female ; Humans ; Male ; Middle Aged ; Pain Management ; Pain, Postoperative ; etiology ; therapy

Objective To evaluate the selection and outcome of procedures for the treatment of female patients with low urovaginal fistulas.Methods Between Jan.1999 and Dec.2012,a total of 94 low urovaginal fistula patients with mean age 28(5-58)years and the duration of the condition for mean 4(1-23)years were treated using a variety of procedures.Of the 94 patients,the etiology was trauma in 57 patients,iatrogenic injuries in 34,local inflammation in 2 and congenital in 1.Urethral stricture was associated with urethrovaginal fistulas in 61 patients(Group of urethra)and vesicovaginal fistula in 33 (Group of bladder).Of the group of urethra,it was associated with ileovaginal fistula in 2 patients,rectovaginal fistula in 3 and veginal strictures in 8.Of the group of bladder,the fistula was simple or incipient in 15 cases and complex or recurrent in 18 cases.In group of urethra,pedicle labial skin grafs urethroplasty was used in 30 cases,island flap of vulva urethroplasty in 4 cases,vaginal wall flap urethroplasty in 18 cases,end to end anastomotic urethroplasty in 6 cases,and anterior bladder flap uretbroplasty in 3 cases.At the same time of urethroplasty,bladder neck reconstruction was performed in 6 cases with pre-existing traumatic sphincter incompetence,intestovaginal fistula repair was performed in 5 cases.In the group of vesicovaginal fistula,the fistula repairing was performed by transabdominal approach in 18 cases and by transvaginal approach in 15 cases.Results There were no serious complications postoperatively.Patients were followed up with mean 45(5-140)months.Of the 34 cases underwent pedicle labial or an island flap of vulva urethroplasty,fistulas was recurrent in 3 cases and urethral strictures was happened in one case,3 cases had frequent and stress incontinence,however,all 3 cases achieved urinary continence in 3-8 months postoperatively;of the 18 cases underwent urethroplasty using vaginal flap repairs,all patients achieved anatomical success,and continence in 15 patients and stress incontinence in 3 patients;of the 6 patients underwent urethral end to end anastomosis,all patients achieved urethral anatomical repair success and the continence was achieved in 5 patients,stress incontinence in one patient;and the left 3 patients underwent bladder anterior wall urethroplasty,urethral anatomical repair success was in all 3 patients and continence in one,stress incontinence in one and dysuria in one.Endoscopic resection was performed in the patient with voiding difficulty,after which the patient could void smoothly.In the group of urethra,successful urethral anatomical recovery rate was 91.8％(56/61)and successful functional recovery rate was 83.6％(51/61).In the group of bladder,the fistulas were successfully repaired by one procedure in 29 patients(87.9％)and recurrence in 4 patients.Conclusions The selection of procedures for treatment of female patients with low urovaginal fistulas should be determined by stricture characteristics,including location,length and vaginal condition.It is very important to prevented fistulas reformation during operation by using tissue flaps as a bulk.

Objective To assess the value of multi-slice computed tomography angiography (MSCTA) in the preoperative detection of aberrant hepatic arteries in patients scheduled to undergo pancreaticoduodenectomy.Methods Patients with pancreatic and peri-ampullary tumors were preoperatively studied using contrast-enhanced abdominal CT angiography (CTA).The results on hepatic arterial anatomy were compared with those obtained from digital subtraction angiography (DSA) and on surgical findings.Results Eighty-one patients were included into this study.DSA was carried out in 29 patients to evaluate tumor resectability,and 66 patients received surgery.Anomalous hepatic arteries were detected in 17 (21％) patients on CTA.Thirteen (16.0％) patients had a single arterial variant,and 4 (4.9％) patients had two arterial variants.One patient each was seen in the Michels type Ⅳ,Ⅶ,and Ⅷ respectively,while 2 patients each were seen in the Michels type Ⅲ and Ⅴ respectively.Five patients were diagnosed as the Michels type Ⅵ,and four as the Michels type Ⅸ.One patient demonstrated a rare variant which was not included into the Michels classification.MSCTA had an accuracy of 100％,a sensitivity of 100％,and a specificity of 100％.Regarding the traceability scores of hepatic arterial segment,there were no statistically significant differences between MSCTA and DSA.Conclusions MSCTA is an effective imaging tool to assess arterial anatomical variation around the pancreatic head.It is non-invasive,and it provides valuable information on the peri-pancre atic vascular anatomy before pancreaticoduodenectomy.

Objective To investigate the feasibility of using small intestinal submucosa (SIS) graft for the repair of anterior urethral strictures. Methods From June 2009 to August 2010, 18 men (mean age, 38 yrs) with anterior urethral strictures underwent urethroplasty using a four-layer SIS as an onlay patch graft. SIS was used to augment the urethral caliber at the stricture site. The mean stricture length was 4.6 cm (range 3.5 to 7 cm). The pre-operative mean maximal flow rate was 3.8 ml/s (range 1.5 to 5.5 ml/s). The required SIS grafts (4 to 7.5 cm long and 2 cm wide) were positioned into the urethrotomy defect and were spread-fixed to the corpora cavernosa using 5-0 polyglactin interrupted sutures. Two apices of the graft were sutured to the proximal and distal apices of the urethrotomy with 5-0 polyglactin interrupted stitches. The margins of the opened urethra were sutured to the SIS patch with 5-0 polyglactin running sutures. Results The mean follow-up period was 10 mon. (range 6-18 mon.). No postoperative complication, such as infection or rejection related to the use of heterologous graft material was observed. Seventeen patients voided well postoperatively with the mean peak urine flow of 25.4 ml/s (14-44 ml/s). Cystoscopy revealed that at four weeks and six weeks, the SIS graft was well distinguishable from the normal surrounding tissue; and at 16 weeks, the urothelium was regenerated and the biomaterial was not distinguishable from the normal surrounding tissue. The squamosal epithelium was seen in the histological examination of the grafts. The remaining one patient with failed hypospadias developed a slight urethral narrowing at five months post-operatively and needed sound dilatations. Conclusions SIS matrix appears to be a safe and effective reconstructive material in selected urethral reconstructions.

Objective To evaluate the long-term efficacy of using penile skin flaps for urethroplasty in the treatment of anterior urethral strictures.Methods Between Jan 2006 and Dec 2012,138 patients with anterior urethral stricture were treated by using penile skin flaps for urethroplasty.The mean age was 38 years (range,7-82 year).The etiology of stricture included trauma in 78 cases,iatrogenicity in 41 cases,infection in 17 cases,unknown reason in 2 cases.The penile urethral stricture was found in 110 cases and the bulbourethral stricture was found in 28 cases.The mean length of anterior urethral stricture was 6.5 cm (range 3-14 cm).Among them,the length of urethral stricture was more than 10 cm in 48 patients.Basing on location,length of stricture and condition of penile skin,different penile skin flaps were chosen,including vertical pedicle skin flap,pedicle circular flap,L-flap,Q-flap.Three different techniques were used for urethroplasty,such as lateral patch flap urethroplasty (group1,n=80),dorsal and ventral inlaid flap urethroplasty (group 2,n =42) and tubularized flap urethroplasty (group 3,n =16).Results 4 patients were lost during follow-up.The mean duration of follow-up in the remaining 134 patients was 39 months (range,8-84 months).Complications developed in 29 of 134 patients (21.6％),including strictures recurrence in 17 (group 1,n =12,group 2,n=2 and group 3,n =3),urethrocutaneous fistulas in 7 (group 1,n =5,group 2,n=1 and group 3,n =1) and urethral diverticulum in 5 (group1,n =4,and group 3,n =1).105 cases voided well and the urinary peak flows ranged from 13-49 ml/s (mean 25 ml/s),The overall success rate was 78.4％ (105/134).Conclusions Penile skin is thinner,rich in blood supply and easy to be manipulated,which is one of the excellent materials for the urethral reconstruction.Q-flap or L-flap urethroplasty is an effective technique for the treatment of long-segment urethral strictrues (≥ 10 cm).