Objective To explore selection of the procedures in one stage urethroplasty for treatment of the coexistence of urethral strictures in anterior and posterior urethra.Methods Between January 2008 and December 2014, a total of 27 patients with coexist strictures simultaneously in anterior urethra and posterior urethra were treated in our hospital.The mean age was 38 years old (ranging 13-83 years old.Stricture etiology was secondary to lichen sclerosus in 2 patients, iatrogenic in 3 and posttraumatic in 22.The mean length of urethral stricture was 11cm (ranging 6-14cm).Two procedures for treatment of anterior urethral stricture, including augmentation of urethroplasty using penile skin flap was performed in 20 patients and augmentation of urethroplasty using lingual mucosa in 7.Three procedures for treatment of posterior urethral stricture, including non-transecting spongiosum end to end anastomosis of the two urethral ends was performed in 3 patients, end to end anastomosis of the two urethral ends was performed in 17 and substitution urethroplasty using different tissues was performed in reminder 7 patients.Of them, pedicle scrotal skin urethroplasty was performed in 2 patients and lingual mucosal graft urethroplasty in 5 patients.Results The patients were mean followed up 2.6 years (ranging 0.545.0 years) with an overall success rate of 88.9％ (24 of 27 cases).Complications developed in 3 patients (11.1％).Of the 17 patients with end to end anastomosis, urethral stricture developed respectively 4 and 6 months in 2 patients and voiding well after pedicle scrotal skin urethroplasty.Urethral pseudodiverticulum developed 9 months after pedicle penile flap urethroplasty in another patient and voiding well after urethroplasty.Urethrography showed patent urethra with adequate lumen in the remaining patients and mean urinary peak flows was 21.3 ml/s (ranging 14.2-37.9 ml/s).Conclusions Substitution urethroplasty using penile skin or oral mucosa was more good procedure for anterior urethral stricture during the treatment of the coexistence of urethral stricture in the anterior and posterior urethra.The treatment of posterior urethral stricture was based on the length of the stricture, local condition to make a choice between anastomotic urethral reconstruction and substitution urethroplasty using other tissue.

Objective To observe the changes of blood sugar following use of recombinant human growth hormone (rhGH) in patients with severe burn. Methods A total of 140 patients with severe burn but with no history of diabetes were randomly and equally divided into Group A ( 18-44 years old) and Group B ( ≥45 years old) according to WHO criteria. Then, two groups were randomly divided Group B0 (treated with normal saline), 35 patients per group. Change of blood sugar and amount of insuline for control of blood sugar were observed and analyzed. Results Of 140 patients in four groups,128 patients accomplished the test, with no statistical difference upon general conditions between groups (P ＞0.05 ). Within four weeks after test, the amount of insuline used for control of blood sugar was (2908.0 ± 153.5) IU and (724.1 ± 31.1 ) IU respectively in Groups A1 and A0, (3725.9 ± 88.4) IU and (801.8 ±22.2) IU respectively in Groups B1 and BO. The results showed that the patients needing insulin to control blood sugar in Group B were more than that in Group A ( P ＜ 0.01 ). The time for blood sugar recovering to normal range in patients with high blood sugar with no use of rhGH in Group A1 and Group B1 was (5.21 ±0.64) d and (5.03 ± 1.01) d respectively (P＞0.05). Conclusions The blood sugar of patients with severe burn will rise after use of rhGH. The age of the patients is positively correlated with changes of blood sugar. Standard use of rhGH does not induce diabetes.

Objective: To establish a method for simultaneous determination of calceolarioside B and chlorogenic acid in Caulis Stauntoniae Chinensis tablets by HPLC.
Methods: The analysis was performed on a Thermo BDS HYPERSIL C₁₈ column (4.6 mm×250 mm,5 μm) maintained at 35 ℃. The mobile phase was consisted of methanol and 0.1% phosphoric acid solution, and gradient eluted with a flow rate of 1.0 mL·min^-1. The detection wavelength was 327 nm, and the injection volume was 10 μL.
Results: The linear ranges of calceolarioside B and chlorogenic acid were 0.51-20.60 μg·mL^-1 (r=1.000) and 0.52-20.63 μg·mL^-1 (r=1.000), respectively. The average recoveries were 100.3% with RSD as 1.1% and 105.9% with RSD as 1.4%, respectively. The content results of 5 batches of Caulis Stauntoniae Chinensis tablets were 0.083-1.115 mg·g^-1 for calceolarioside B and 0.161-1.204 mg·g^-1 for chlorogenic acid.
Conclusion: The method can be used for improving the quality evaluation standard of Caulis Stauntoniae Chinensis tablets.

Objective To discuss the curative effect of different operative methods for the treatment of urethrocutaneous fistula (UCF) after urethroplasty.Methods Clinical data of 54 cases of UCF from January 2003 to July 2011 were collected.Simple suture,advancement skin flap,pedicle penile skin flap urethroplasty,tongue/buccal mucosa urethroplasty were performed according to the size,location,number and whether there was urethral stricture of UCF.The treatment effect was recored and analyzed.Results The success rate of UCF repair was 85％ (46/54).There were 4 patients cured after second surgery.The success rates of repair of UCF with simple suture,advancement skin flap,pedicle penile skin flap urethroplasty,tongue/buccal mucosa urethroplasty were 95％,76％,82％,and 83％,respectively.Postoperative recurrence was observed in 5 patients in 6 months after the surgery and 4 of them received the re-operation.Conclusion According to the principle and the individual circumstance,taking personalized operative methods may improve the success rate of the repair of all kinds of UCF.

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 clone and express Dengue virus nonstructural protein 4A (NS4A) gene and express in eukaryotic cells.Then,to isolate and purify and isolate cellular proteins interacted with NS4A.Methods With specific primers,NS4A gene fragment tagged with FLAG and HA (FLAG-NS4A-HA) was amplified by PCR and cloned into an expression vector,pSG5 vector.Recombinant plasmid was transfected into A549 cells by LipofectAMINETM2000.Transient expression of FLAG-NS4A-HA was detected by Western blot.The NS4A interacting proteins were isolated and purified by tandem affinity purification (TAP) system using HA and FLAG antibodies,and then assayed by silver stained SDS-PAGE.Results Dengue virus NS4A gene tagged with FLAG and HA was successfully constructed into pSG5 vector and expressed in A.549 cells.Silver stained SDS-PAGE showed that the expressed NS4A and two potential interacting proteins that interact with NS4A were isolated after TAP purification and SDS-PAGE.Conclusion Cellular proteins that potentially interacted with Dengue virus NS4A were successfully purified and isolated,which provided a basis for further research.

Objective To establish a tandem affinity purification(TAP) system of innate immune-regulatory protein PKR and analyze PKR function, for the future screen and identification of novel PKR-interaction proteins. Methods PKR gene was amplified by PCR, and then cloned into a mammalian expression vector pcTAP-A. Recombinant pcTAP-PKR was transfected into PKR knock-down(PKRkd) HeLa cells by LipofectAMINE 2000,and the PKR overexpressed HeLa cells were harvested for mitogen-activated protein kinases(MAPK) activation analysis. Cell extracts of PKR overexpressed cells were purified using TAP kit and examined by Western blot. Results Cal modulin resin(CBP) and streptavidin resin(SBP) tagged PKR was detected in PKRkd HeLa cells as early as 24 h upon transfection with pcTAP-PKR, and its expression decreased at later time points. The overexpression of PKR was autophosphorylated, and thus involved in the regulation of MAPK actviation. After small-scale TAP kit purification, PKR protein was detectable by Western blot. Conclusion We have successfully established a TAP system that over-expresses functional PKR, providing a useful tool for the future study on the identification of PKR interacting proteins.

Objective To investigate the differentiation of rat bone marrow mesenchymal stem cells (MSCs) to renal tubular epithelial-like cells under different conditions. Methods MSCs were obtained from rat marrow. MSCs were isolated by gradient density centrifugation and plastic adherence and then purified. Surface markers were identified with flow cytometry after amplification in vitro. The purified MSCs of the third passage were cultured respectively as follows: (1) control group: DMEM medium with fetal bovine serum(FBS). (2) all-trans retinoic acid (ATRA) group: DMEM medium with FBS, ATRA and ischemic reperfusion-injured kidney tissue homogenate. (3)combination group: DMEM medium with FBS, ATRA, ischemic reperfusion-injured kidney tissue homogenate, epidermal growth factor (EGF) and bone morphogenetic protein 7 (BMP-7). After 7 days, the MSCs were collected for alkaline phosphatase (AKP) staining, cytokeratin-18 and E-cadherin immunocytochemical analysis. Results The positive rates of the third passage MSCs in CD44, CD90 and CD29 were 97.8%±0.9%, 96.8%±1.4% and 97.6%±2.4%,respectively, but in CD11b/c and CD34 were only 13.2%±0.6% and 1.2%±0.5%. The MSCs in control group were spindle. The MSCs in ATRA group were round and elliptic. The MSCs in combination group became cobblestone-like cells after 7 days. AKP staining showed that tubular epithelial-like cells from MSCs in control group were negative, some above cells in ATRA group were positive and number of above cells increased in combination group. Compared with negative control group, the ratios of cytokeratin-18 positive cells in ATRA group and combination group were respectively increassed by 29.47%±1.08% and 47.52%±2.13% (all P<0.05), the ratios of E-cadherin positive cells in ATRA group and combination group were respectively increased by 14.88%±2.46% and 36.15%±1.13% (all P<0.05). Conclusion MSCs may differentiate by renal tubular epithelial-like cells under the induction of ischemic reperfusion-injured kidney tissue homogenate and ATRA in vitro, which are further differentiated under the combined induction of EGF and BMP-7.

ured with different techniques.Standardized application of somatropins will not induce diabetes.

Objective To investigate the efficacy and safety of using combined lingual mucosa and buccal mucosa onlay grafts or foreskin flap urethroptasty for the treatment of long or multi-seg-ment urethral strictures. Methods Seven patients with long and 4 cases with multi segment urethral strictures(range 10 to 15 cm,mean 12)underwent substitution urethroplasty using combined lingual mucosa and buccal mucosa onlay grafts or foreskin flap urethroplasty.The patients'age ranged 24 to 56,mean 32 and the course of disease was from 6 to 96 months.Of the 11 patients 7 underwent com-bined lingual mucosa and buccal mucosa grafts urethroptasty,4 patients underwent combined lingual mucosa graft and foreskin flap Urethroplasty. Results The patients were followed up 5-1 2(mean 10)months postoperatively. Meatal stenosis developed 3 months postoperatively in 1 patient who un-derwent combined lingual mucosa and foreskin flap urethroplasty.The patient could void well after re-operation.The other patients could void well and the peak flow rate ranged from 2 1 to 3 6 ml/s(mean 26.8 ml/s). Conclusions Combined lingual mucosa and buccal mucosa onlay grafts or foreskin flap substitution urethroplasty may have the advantage of easier harvest,less trauma.It could be a good U- rethral substitution technique for the treatment of long or multi-segment urethral stricture.