BACKGROUND:Many studies have shown that different nanostructures produce different influences on cellbioactivity, but the nanonetwork structure is not reported yet. OBJECTIVE:To study the influence of the nanonetwork topography on the bioactivity of bone marrow mesenchymal stem cels. METHODS:The nanonetwork topography was fabricated on biomedical titanium surface by alkali-heat treatment, and pure titanium served as control group. Bone marrow mesenchymal stem cels were co-cultured with the above two types of samples. cellmorphology and cytoskeleton were observed using scanning electron microscope and immunofluorescence method. The celladhesion, proliferation and osteogenic differentiation were detected by measurement of absorbance values at different culture time. RESULTS AND CONCLUSION: The nanonetwork topography had significant advantage on the number of adherent cels at 30, 60 and 120 minutes of co-culture. The cellproliferation was significantly accelerated by the nanonetwork topography at days 1, 3, 5 of co-culture, and the absorbance values in the nanonetwork group were significantly higher than those in the pure titanium group (P < 0.05). The alkaline phosphatase activity in the nanonetwork group was also significantly higher than that in the pure titanium group at 14 days of osteogenic induction (P < 0.05). The cellshape and cytoskeleton on the nanonetwork surface were better than those on the titanium surface. These findings indicate that the nanonetwork topography has better effects on cellbioactivity compared with pure titanium.

BACKGROUND:Autologous cartilage has a poor self-repair effect due to low chondrocyte density, low metabolism rate and no blood supply.
OBJECTIVE:To summarize the recent study about tissue engineering techniques and surgical treatment for cartilage injury.
METHODS:A computer-based online retrieval of PubMed database was performed by the first author for articles published between January 1992 and December 2013. The key words are“articular cartilage, injury, tissue engineering, repair”in English. According to the inclusion and exclusion criteria, 61 literatures were included into the final analysis.
RESULTS and CONCLUSION:The current clinical treatment of articular cartilage injury includes joint debridement, mosaicplasty, perichondrium transplantation and autologous chondrocyte implantation. However, their long-term result is unsatisfactory. One reason for limited clinical success is that new cartilage can be formed at the site of a defect, and the repaired tissue canot compare with the autologous cartilage in mechanical property. Tissue engineering technique is stil a hot topic in recent years, because it can potential y induce autologous cartilage formation. Through endogenous or ectogeneous seed cells and inducting factor and nutrient factors, tissue engineering technique can be applied to induce the self-repair of articular cartilage, thus regenerating into hyalinc cartilage with the similar even same biological property. How to simplify the treatment protocols and reduce treatment cost is the key to promote cartilage repair.

BACKGROUND: There are a lot of immunologic studies about limb allotransplantation in animal experiment. But, it is only early investigation in clinic; its clinical immunologic study needs further accumulation.OBJECTIVE: To dynamically analyze the early immunologic state change in patients following hand allotransplantation.DESIGN: Controlled trial.SETTING: Department of Orthopaedics, Guangzhou General Hospital,Guangzhou Military Area Command of Chinese PLA; Department of Traumatic Orthopaedics and Department of Laboratory Medicine, Nanfang Hospital, First Military Medical University of Chinese PLA.PARTICIPANTS: Two patients who underwent unilateral hand allotransplantation in the Department of Orthopaedics, Guangzhou General Hospital,Guangzhou Military Area Command of Chinese PLA were enrolled, serving as experimental group. The observation was between September 1999 and March 2000. Twenty persons, including 12 male and 8 female, who homochronously received health examination, aged 20 to 45 years, were enrolled, serving as healthy control group. They all had no reactive immune and infectious diseases, and voluntarily participated in the trial.METHODS: Peripheral blood was collected from 2 patients who underwent hand allotransplantation once respectively at pre-operative 1 day and 3 days. Blood collecting was performed once per day at post-operative 1 week, three times per week at post-operative 2 to 4 weeks, twice per week at 5 to 8 weeks post-operation, once per week at 9 to 16 weeks post-operation, twice per month at 5 to 6 months post-operation. ① Peripheral blood T cell subgroups (CD3+,CD4+,CD8+T cells)were detected by flow cytometer,serum panel reactive antibody (PRA) by ELISA method, serum C-reactive protein by turbidimetric immanoassay (TIA), serum creatine kinase (CK) by enzyme dynamics method. ②Mixed lymphocyte reaction(MLR): mitomycin C-treated donor peripheral blood lymphocytes were used as stimulator, and proliferative reaction of peripheral blood lymphocyte of patients to donor transplanted antigen was detected with the incorporation of 3H-TDR method (Negative: There was no significant difference between the mean value of stimulation index and 1, conversely positive). Autogenic peripheral blood lymphocytes treated with the same way replaced donor stimulator, serving as control. Stimulation index of each specimen was calculated (Stimulation index=Experiment cmp/controlcpm), serving as control index. Peripheral blood T-cell subgroups (CD3+,CD4+,CD8+T cell), serum PRA, C-reactive protein and CK were detected in 20 persons in healthy control group;Twenty persons were randomly divided into 10 groups. Two persons in each group were used as donor and recipient mutually and performed MLR.MAIN OUTCOME MEASURES: ① Peripheral blood T cell subgrpups (CD3+,CD4+,CD8+T cell). ②PRA. ③ C-reactive protein. ④CK. ⑤MLR.RESULTS: ①CD3+,CD4+,CD8+T cell levels were obviously decreased within one week after operation. CD3+ and CD4+T cell levels both recovered to be the pre-operative levels, but CD8+ level exceeded pre-operative level significantly [CD3+: (66.43±4.56); CD4+: (30.55±3.94); CD8 +:(33.45 ±2.69)]. There was no significant difference between experiment group and control group. ②Serum PRA was 0 to 10%, there was no significant difference as compared with control group. ③ Serum C-reactive protein was 0 to 0.359 mg/L, there was no significant difference as compared with control group. ④ Serum CK was 25 to 170 mmol/L, and there was no significant difference as compared with control group. ⑤ MLR after transplantation was negative, and it turned into be positive 5 months later.They were all positive in control group.CONCLUSION: Short-term change and long-term redistribution of T cell subgroups are closely related to immunosuppressive agent, suggesting that immunosuppressive agent has obvious effect on T-cell subgroup following hand allotransplantation. Immuno-induction schedule make patients be in immune suppression state, which effectively avoid early rejection. But patients cannot bear specificity yet; they need the inhibition of immunosuppressive agents.

BACKGROUND: There are many methods for posterior cruciate ligament (PCL) reconstruction, which is involved in many graft materials, but few studies aim to compare the differences in outcomes of different grafts for PCL reconstruction. OBJECTIVE: To compare the clinical results of arthroscopic PLC reconstruction with bone-patellar tendon-bone (B-PT-B) autograft, B-TP-B allograft and semitendinosus tendon autograft. DESIGN, TIME AND SETTING: A retrospective case analysis was completed in the Department of Orthopedics, Guangzhou General Hospital of Guangzhou Area Military Command of Chinese PLA from January 2000 to September 2005. MATERIALS: Totally 76 patients underwent arthroscopic PLC reconstruction from January 2000 to September 2005, with the use of B-TP-B autograft in 21 patients, B-TP-B allograft in 27 patients, semitendinosus tendon autograft in 28 patients. METHODS: A retrospective analysis was performed in 76 patients underwent arthroscopic PCL reconstruction, with the use of B-TP-B autograft in 21 patients, B-TP-B allograft in 27 patients, semitendinosus tendon autograft in 28 patients. Postoperative body temperature was examined duration hospitalization. The follow-up parameters included International Knee Documentation Committee (IKDC) scores, Lysholm knee joint scores, and KT-1000 evaluation.MAIN OUTCOME MEASURES: ①Range of motion. ②joint stability: posterior draw test and KT-1000 test. ③overall function of knee: IKDC scores and Lysholm scores; ④complications and side effect. RESULTS: The time of follow-up visit was 26-79 months. Differences were no statistically significant among the IKDC scores, Lysholm scores, KT-1000 side-side difference, the positive rate of posterior draw test in three groups of patients with PCL reconstruction using B-TP-B autograft, B-TP-B allograft and semitendinosus tendon graft (P > 0.05); 10° flexion limitation was found in 3 cases of B-TP-B autograft, 5° flexion limitation in 1 case of B-TP-B allograft and flexion limitation in 2 case of semitendinosus tendon graft. There was no significant difference in the ratio of knee joint flexion limitation among three groups. No synarthrophysis, wound infection, implant disrupture, screw loose, patellar fracture or vascular nerve injury was observed in three groups of patients; There were 12 cases presenting anterior knee pain in the B-TP-B autograft group and 5 cases presenting posterior knee pain in the semitendinosus tendon graft group. The difference of peal-knee pain incidence was statistically significant among three groups (P=0), the highest in B-TP-B autograft group, then semitendinosus tendon graft group and the lowest in B-TP-B allograft group. The time of post-operative fever in B-TP-B autograft group was earlier than that in the B-TP-B allograft and semitendinosus tendon graft groups (P=0). There was no significant difference between allogreft group and semitendinosus tendon autograft group (P=0.844). The rejections appeared in 4 cases of B-TP-B allograft with the manifestations of the sustained jam-like liquid outflow from tibial tunnel. After dressing, hormones or indomethacln, the rejection was healed. CONCLUSION: The arthroscopic B-TP-B autograft, B-TP-B autograft and semitendinosus tendon autograft have the same clinical curative effect in PCL reconstruction.

Objective To analyse the operation technique and therapeutic effect of "inserting" uretero-intestinal anastomosis in orthotopic bladder substitution.Methods Thirty-eight patients undergoing orthotopic bladder substitution operations were followed up,and the way of uretero-intestinal anastomosis in all Datients was the "inserting"uretero-intestinal anastomosis.The therapeutic effect was observed by radiation,cystoscopy,pathologic biopsy and blood test.Results The average follow-up time was(3 1.65±14.14)montll8.and the stricture rate was 4%(3/75),but no vesicoureteric reflux was found.The rate of leakage was 0.Nipples were formed at the site of anastomosis under the view of cystoscope,and among the 7 patients whose nipples were taken to be examined by histology,2 cases were intestinal epithelium which were taken at the base of nipple8.while the others were transitional epithelium which were taken at the top of nipples.The renal function of all patients was normal (Cr 54-135 μmol/L,BUN 3.2-9.4 mmol/L).Conclusion "Inserting"uretem-intestinal anastomosis is an ideal antireflux uretero-intestinal anastomosis method.

Objective To evaluate the efficacy of mitomycin (MMC) intravesical chemotherapy for superficial bladder carcinoma by in vitro chemosensitivity using histoculture drug response assay (HDRA).Methods Forty-one cases of superficial bladder transitional cell carcinoma(TCC) were obtained,including 30 males and 11 females with mean age of 55 years.Of them,10 cases were Ta and 31 were T1 according to TNM stage system (UICC 2002) while 9 cases were G1,22 were G2 and 10 were G3 (WHO1973).All cases had no chemotherapy history before operation and were operated to retain bladder.Tumor specimens were cultured by three-dimensional histoculture.HDRA with im-proved MTT assay was utilized for chemosenstivity test of MMC with 1 g/L concentration and 2 hours exposure.Growth inhibition rate (GI) exceeding 70% was defined as high-sensitive while less than 50% GI was defined as insensitive,others were moderate-sensitive.All cases were performed standard intravesical chemotherapy with MMC 40 mg plus 40 mt saline.Every case was followed up every 3 months.The patients were followed up for 5 years or untill recurrence.Results The MMC chem-osensitivity was different among 41 patients.Thirteen cases were insensitive including 1 of Ta,12 of T1 (P=0.009) and 1 of G1,7 of G2,5 of G3(P=0.101).Total recurrence rate was 39%(16/41) af-ter 3 to 5 years follow-up.There were 1 of Ta,15 of T1 (P=0.059) and 10 of G2 6 of G3 (P=0.016).Insensitive group recurrence rate was 77% (10/13) while sensitive group was 21% (6/28,P= 0.004).Patients in sensitive group showed a longer median time(49.2 months) than patients in insen-sitive group (16.5 months,P<0.001) according to Kaplan-Meier analysis with Log-rank test.The MMC chemosensitivity was independent prognostic factor examed by Cox regression analysis (P= 0.008).There was a 78% correlation rate of chemosensitivity by HDRA to clinical effect of MMC in-travesical chemotherapy.Conclusion HDRA could evaluate MMC intravesical chemotherapy for su-perficial bladder TCC,improve therapeutic effect and lower tumor recurrence rate.

Objective To study the application of pediculated skin flaps in the treatment of com-plicated long urethratresia. Methods From March 1999 to May 2006, a total of 18 male patients with complicated long urethratresia were treated by using the pediculated skin flaps. The causes of urethratresia were 7 cases of postoperative pelvic fractures with posterior urethral stricture, 4 cases of transurethral intravesical chemotherapy, 3 cases of postoperative bulbar urethral stricture, 2 cases of gonorrhea, and 2 cases of long-time urethral catheter placement. Four cases were urethratresia nf cor-pus penis, 7 cases were anterior urethral obliteration, 7 cases were posterior urethral and anterior ure-thral obliteration. Urethro-perineal fistulas were found in 8 cases, posterior urethrorectal fistulas in 7 cases, false passage formations in 8 cases. The average length of urethratresia was 15.1 cm (range 8. 7 to 23. 0 cm). The urethral scar was rasected, the posterior urethrorectal fistula was repaired, and different kinds of pediculated skin flaps depending on the length of urethratreaia was used. Results All the patients were followed up for 12 to 18 months (mean 14 months). Fifteen patients voided well 3 months postoperatively, none of the urography showed stricture. The mean peak urinary flow rate was 16. 9 ml/s (range from 16. 5 to 21.7 ml/s). Of the other 3 cases, 1 case experienced difficult voi-ding due to the long and circuitous tabularized skin flap but recovered after proper shortening;1 case had restenosis for the infection of anastomosis but voided well after excision and reanastomnsis;1 casehad a urinary fistula resulting from hematoma and infection, but was successfully treated by the neo-plasty of the urinary fistula. The mean peak urinary flow rate was 17.0 ml/s (range 15.0 to 22.0 ml/s) for 17 patients 6 months postoperatively, except for one who experienced genuine urinary incon-tinence. At 9-18 months after operations, the mean peak urinary flow rate was 17.5 ml/s (range 15.8 to 22.5 ml/s) for 17 patients. Conclusion The single-stage urethroplasty based on pediculated skin flaps is a reliable and durable method for complicated long urethratresia.

Objective: To explore the effects of various forms of prostatic apex on positive apical margin rate (PAM) and biochemical recurrence (BCR) after laparoscopic radical prostatectomy. Methods: A retrospective analysis of 309 patients (aging (65±6) years) who were experienced laparoscopic radical prostatectomy from January 2010 to December 2016 at the Department of Urology, First Affiliated Hospital of Fujian Medical University. According to the relationship between prostate apex and membrane urethra at the mid-sagittal plane of preoperative MRI, all patients were classified into 4 categories. There were 31 patients for type 1, apex covering both anterior and posterior aspects of membranous urethra, 139 patients for type 2, apex covering anterior side of membranous urethra, 63 patients for type 3, apex covering posterior aspect of membranous urethra, 76 patients for type 4, apex not covering membranous urethra. PAM and BCR after operation were compared between this four groups respectively. The χ² test was used to compare PAM among the 4 types. Logistic regression analysis were undertaken to analyze the factors affecting PAM. Cox′s proportional hazards regression model was undertaken to identify the variables influencing BCR. Results: There was no significant difference in the 4 groups concerning age, body mass index, prostate volume, preoperative prostate-specific antigen (PSA) value, postoperative Gleason score and pathological stage (P>0.05).The median follow-up time was 32 months (ranged from 12 to 60 months).The data showed that the apical type 3 patients has the highest PAM. There was statistical difference among the 4 groups in PAM (χ²=15.592, P=0.001). Preoperative level of PSA (OR=20.356, 95% CI: 2.440 to 169.810, P=0.005), postoperative Gleason score (OR=4.113, 95% CI: 1.911 to 8.849, P=0.001), pathological stage (OR=3.422, 95% CI: 1.600 to 7.319, P=0.002) and apical type 3 (OR=6.134, 95% CI: 2.196 to 17.132, P=0.001) were independent relactive factors of PAM. Preoperative level of PSA (HR=1.362, 95% CI: 1.006 to 1.843, P=0.045), postoperative Gleason score (HR=1.920, 95% CI: 1.384 to 2.665, P=0.001), pathological stage (HR=1.476, 95% CI: 1.098 to 1.983, P=0.010), PAM (HR=3.497, 95% CI: 2.407 to 5.081, P=0.001)and apical type 3 (HR=1.828, 95% CI: 1.266 to 2.639, P=0.001) were independent prognosis factors of BCR. Conclusion Prostate apical type 3 could be a significant independent predictor of PAM, and an independent prognosis factor for BCR.

AIM:To investigate the role of Rho-associated kinase ( ROCK) inhibitor fasudil in the formation of rabbit urethral stricture after injury and to observe the cell activity , migration and extracellular matrix synthesis in the rabbit urethra fibroblasts.METHODS:The rabbit model of urethral stricture was established by microsurgical techniques .The rabbits were divided into sham operation group , operation group and fasudil (3 mg/kg, 10 mg/kg, 30 mg/kg) groups.The diameter of the stenosis was measured by retrograde urethrography 3 months after surgery .The fibroblasts were isolated from urethral scar, and then incubated with fasudil (12.5 μmol/L, 25 μmol/L, 50 μmol/L) in the presence of transforming growth factor-β1 (TGF-β1, 10 μg/L).The untreated cells were used for control .The cell activity was measured by MTT assay.The cell migration ability was tested by the method of Transwell chambers .The protein expression of ROCK , α-smooth muscle actin (α-SMA) , collagen I and collagen III was determined by Western blot analysis .RESULTS:Fasudil significantly reduced formation of urethral stricture after injury (P<0.05).Cultured rabbit fibroblasts with different con-centrations of fasudil inhibited the cell activity and cell migration ability (P<0.05).The protein expression of ROCK,α-SMA, collagen I and collagen III was also inhibited by treatment with fasudil in a dose -dependent manner ( P<0.05 ) . CONCLUSION:Fasudil inhibits the formation of extracellular matrix and reduces the incidence of urethral stricture after injury by down-regulating TGF-β1-induced Rho/ROCK pathway activation in the rabbit urethra fibroblasts .

Objective To evaluate the predictors of gleason score pathological downgrading after radical prostatectomy in patients with biopsy-proven level 2 of grading groups (Gleason Score 3 + 4 =7).Methods Data of 252 patients,diagnosed with level 2 of grading groups(Gleason score 3 + 4 =7) prostate cancer by biopsy,with subsequent laparoscopic radical prostatectomy,were retrospectively analyzed.The mean age was 64.3,ranged from 46 to 82 years.The average body mass index (BMI) was 23.2 kg/m2,ranged from 15.2 to 30.4 kg/m2.The median prostate volume,transition zone volume(TZV) and transition zone index(TZI) were 48.9 ml (30.3-73.1 ml),21.4 ml(13.5-31.2 ml) and 0.46％ (0.37％-0.58％),respectively.The median value of tPSA,fPSA and PSAD were 1.53 ng/ml(0.67-3.92 ng/ml),9.65 ng/ml (4.13-18.68 ng/ml) and 0.18 ng/(ml · cm3) [0.09-0.50 ng/ (ml · cm3)],respectively.Clinical T stage was also evaluated,including 153 (60.7％) diagnosed as T1e stage,78 (3 1.0％) diagnosed as T2 stage,and 21 (8.3％) diagnosed as T3 stage.There were 58(23.0％) with extracapsular extension,47 (18.7％) patients with seminal vesicle invasion,and 2(0.8％) with lymph node metastasis.Pathological T stage includes 112 (44.4％) diagnosed as T2 stage,55 (21.8％) diagnosed as T3a stage,35 (13.9％) diagnosed as T3b stage,and 50(19.8％) diagnosed as T4 stage.The patients were assigned Prostate ImagingReporting and Data System version 2 scores of 1,2,3,4,and 5 were 45 (17.9％),36 (14.3％),51 (20.2％),68(27.0％)and 52(20.6％),respectively.The patients were categorized into 2 groups with and without pathological downgrading,including downgrade and no downgrade group.Univariate and multivariate logistic regression analysis were done to determine the predictors of pathological downgrading.Results The patients were categorized into downgrade(n =31) and no downgrade group(n =221) of 252 patients.The pathological downgrading was identified in 31 (12.3％).The tPSA,PSAD and PI-RADS scores in patients with downgrade group which were lower than those in without downgrade group (P ＜ 0.05).The logistic regression analysis revealed PI-RADS score was the independent predictor of downgrading(OR =0.364,95％ CI 0.253-0.522,P ＜ 0.01).The area under the ROC curve of PI-RADS score was 0.810 and the diagnostic value was the best.Conclusions These findings suggested that PI-RADS scores was predictor for pathological downgrading after radical prostatectomy in patients with biopsy-proven level 2 of grading groups,reduced PI-RADS score (PI-RADS score ≤ 3) is correlated with increased pathological downgrading after radical prostatectomy.