BACKGROUND:Sodium butyrate, a histone deacetylase inhibitor, can inhibit cell proliferation, and induce apoptosis and differentiation of various cancer cells. However, the role of sodium butyrate combined with tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) on lung cancer stem cells remains unclear. OBJECTIVE:To explore the effect of sodium butyrate combined with TRAIL on biological behaviors of lung cancer stem cells. METHODS:Magnetic bead separation was used to separate lung cancer stem cells (CD133+) from human lung adenocarcinoma A549 cells. After the lung cancer stem cells were treated with simple DMEM/F12, DMEM/F12 containing sodium butyrate (5 mmol/L), TRAIL (50 μg/L) or sodium butyrate combined with TRAIL, the cell proliferation within 96 hours of culture was determined by MTT assay; the apoptosis within 24 hours of culture was measured by flow cytometry; the cell migration within 48 hours of culture was detected by cell scratch test; the expression levels of pluripotent transcription factors (Oct4, Sox2 and Nanog) within 48 hours of culture were detected using western blot analysis. RESULTS AND CONCLUSION:The CD133+ lung cancer stem cells were successfully enriched from human lung adenocarcinoma A549 cells. MTT assay showed that sodium butyrate and TRAIL significantly inhibited the proliferation of lung cancer stem cells (P< 0.05), and the combination effect was even stronger (P < 0.05). Results from flow cytometry analysis and scratch test showed that sodium butyrate or TRAIL induced apoptosis and inhibited cell migration of lung cancer stem cells (P < 0.05), and the combination of sodium butyrate and TRAIL showed a stronger effect (P < 0.05). In addition, the expression levels of Oct4, Sox2 and Nanog were significantly down-regulated by sodium butyrate (P < 0.05), TRAIL or sodium butyrate combined with TRAIL, and the combination effect was stronger (P < 0.05). In conclusion, sodium butyrate and TRAIL have synergistic effects on lung cancer stem cells, indicating a new way for treatment of lung cancer.

Objective To evaluate the effect of alendronate on osteoprotegerin (OPG) and receptor of activator of nuclear factor κB-ligand (RANKL) expression in human marrow stroma cells (hMSCs) in vitro. Methods hMSCs were isolated from haman marrow, cultured in vitro, and randomly divided into two groups: alendronate group, hMSCs culture fluid containing 1×10-7 mol/ L alendronate; control group, no special treatment but culturing hMSCs in DMEM. Two weeks after treatment, the expressions of OPG and RANKL were evaluated by RT-PCR and Western blot.Results hMSCs became uniform spindle-shaped fibroblasts. As cells proliferated, they formed colonies and showed whirlpool arrangement. After one week's treatment, hMSCs in alendronate group had reduced processes and gradually showed disc shape, which did not happen in control group but kept fibroblast shape and just increased in density. In RT-PCR, the ratio of OPG/RANKL in alendronate group and control group was 8.77±1.16 and 4.58±1.27, respec-tively. In Western blot, the ratio of OPG/RANKL in alendronate group and control group was 2.58±0.47 and 1.52±0.32, respectively. The ratio of OPG/RANKL was higher in alendronate group than in control group (P<0.01).Conclusion Alendronatc enhances OPG expression and inhibits RANKL expression of hMSCs in vitro.

Objective To assess the clinical efficacy of a modified complete retroperitoneal laparoscopic nephroureterectomy via 3 port approach.Methods From August 2013 to February 2016,23 patients with complete retroperitoneal laparoscopic renal and ureteral sleeve resection were treated with modified three port approach,including 15 males and 8 females.The average age was 67 years old (ranging 44-83 years old).All patients had complained about the hematuria before operation and urine exfoliated cells showed moderate to severe nuclear atypia.All patients accepted the abdominal CT and urography CTU examination,pre-operatively.All of them was diagnosed localized upper urinary tract malignant tumors based on those images,including 13 cases in the pelvis,and 10 cases in the upper segment of the ureter.No chemotherapy,radiotherapy or immunotherapy was performed before surgery.No patients have the history of severe basic disease or upper urinary procedure.The operations were performed under general anesthesia,patients take the contralateral back 30 degrees slope,low elevation head foot,waist bridge,side waist stretch.In the anterior superior iliac spine perpendicular to the line 2 cm parallel to the lower intersection of the rib border were disposed into the 12 mm trocar.Above the anterior superior iliac spine two cross finger level with the intersection of the anterior axillary node,we placed into the 10 mm trocar placement lens.Laparoscopic placement of third casing form an isosceles triangle with the first two casing.The renal fascia was incised with an ultrasonic knife from the renal dorsal side,and the renal hilum was isolated from the kidney by suction aspirator.The renal artery and vein were separated and closed by hem-o-lok.Along the psoas muscle surface to ureter,ureteral clipping by hem-o-lok but not to cut off the free distal ureteral,the lens is composed of first casing into,using ultrasonic knife to free ureter to the bladder wall segment,with 30 mm endoscopic stapler ureter and bladder wall cut off part.Operation time,blood loss and postoperative recovery were recorded in 23 cases.Results All 23 cases were successfully operated without related the operative complication.The operative duration ranged from 3.5 to 6.1 h (mean 4.8 h),the blood loss was 30-880 ml (mean 304 ml),and the postoperative stay was 8-30 d (mean 17.8 d).There are 3 cases of positive lymph node by postoperative pathological reports.Within 2 to 30 months following up,2 patients died of tumor progression in 6 months after surgery.4 patients were diagnosed with bladder cancer in 15 months,15 months,21 months,24 months after surgery,respectively.And the transurethral resection of bladder tumor was performed.Conclusion The modified complete retroperitoneal laparoscopic nephroureterectomy via three ports is safe and reliable.

ObjectiveTo explore the association between hepatic steatosis and the liver tissue expression of HBsAg and HBcAg in chronic hepatitis B (CHB) patients.MethodsFrom January 2005 to June 2008,a total of 147 CHB patients with hepatic steatosis diagnosed by liver biopsy and other 149 CHB patients without hepatic steatosis but with similar HBV DNA titer were enrolled.The differences of HBsAg and HBcAg immunostaining and liver injury in these two groups were compared.The data were analysed using t test and chi square test.ResultsCompared with non-steatosis group,the average age and body weight index of hepatic steatosis group were higher (t values were -3.31and -6.57,both P＜0.01).The percentage of moderate to severe hepatic inflammation in liver,obvious hepatic fibrosis and the strong positive HBsAg staining was lower (30.6％ vs 15.4％ ； 26.5％vs 12.8％； 23.1 ％ vs 6.7 ％； x2=9.63,8.92,15.76； all P＜0.01),and the percentage of strong positive HBcAg staining was also in downtrend.Compared with degree F1 and F2 of liver steatosis,the percentage of HBsAg and HBcAg strong positive staining in liver tissues of degree F3 and F4 was in downtrend.ConclusionsHepatic steatosis affected the expression of HBsAg and HBcAg in liver tissue of CHB patients.As hepatic steatosis appeared and became more severe,both expression of HBsAg and HBcAg and the degree of liver injury were in downtrend.

Objective To elucidate whether and how injection site diversity influences on the sodium hyaluronate (HA) distribution on cartilage surface of the knee joint.Methods From September 2014 to December 2014, 32 human cadaveric knees were sorted from small to large based on femoral condyle width (FCW) and divided into group Ⅰ (n=16, odd number, the patella medial approach) and group Ⅱ (n=16, even number, injected through the medial joint line approach).Knees of both groups were interfused with a 5％ (mg: ml) methylene blue tracer.Each specimen was then simulated a 5°-45° alternating walking for 2 h in the Electro Force platform.Afterwards, all the knee cavities were cut open to examine the range of HA distribution.Based on the eight-zoning classification law for the knee cartilage surface, the coverage area of HA in each zone was scored and the characteristics of HA distribution was depicted, as well as HA zonal distribution diversity between groups were statistically analyzed.Results HA of both patella medial and medial joint line subgroups showed analogical distribution in all zones except the lateral tibial plateau at the time of FCW ≤ 7.0 cm.However, HA coverage through the patella medial approach showed significantly higher distribution scores in the patella zone and the femoral trochlear zone, but significant lower distribution scores in the antero-lateral/postero-lateral femoral condyle, the medial posterior femoral condyle zone and the lateral tibial plateau zone compared with medial joint line approach at the time of FCW ＞ 7.0 cm.Conclusion When delivered through different injection approaches, HA showed analogical distribution characteristics in most zones of the knee cavity in population with a small bodily shape (FCW ≤ 7.0cm), but significant diverse distribution characteristics in population with a big bodily shape (FCW ＞7.0 cm).When FCW ＞ 7.0cm, HA injected through the medial-patellar approach tended to be distributed mainly over the patella-femoral articulation and the anterior knee cavity, while HA injected through the joint line approach tended to be distributed mainly over the tibio-femoral articulation and the posterior knee cavity.

Objective To observe the effect of a novel pressure controlled water jet used in transurethral resection of bladder tumor. Methods Clinical data of 12 patients who underwent a transurethral resection of bladder tumor by using a novel water jet was retrospectively analyzed. Results All 12 cases were successfully operated. Estimated blood loss during operation was less than 5 ml. Operation time was 20-45 minutes (median was 28 min), time of water injection was 8-15 minutes (median was 12 min). There were no significant complications. All patients were followed up for 4-10 months, and tumor recurrence or progression was not found by cystoscopic examination. Conclusion Transurethral resection of bladder tumor by using a novel pressure controlled water jet is advantageous with favorable safety and feasibility.

Objective:To establish a pristine-induced rheumatoid arthritis model in mice,and to evaluate its histological and immunological distinction.Methods:Thirty female BALB/c mice,6-8 weeks old,were randomly divided into 2 groups,a control group and pristine group.The mice in pristine group were injected intraperitoneally with 0.5 ml pristine three times at 0,9,and 18 weeks, while mice in the control group receiving saline at the same time.Arthritis score and paw thickness were measured and histopathological assessment of joint sections was performed.The expression of phagocytes,dendritic,neutrophils,T and B cells markers in spleen were determined by flow cytometry.Results:In model-marking group,11 mice were presented with macroscopic evidence of arthritis such as erythema or swelling.The paw thickness in pristine-induced mice was significant higher than that in the control groups[(2.90±0.51) mm vs(1.29±0.47 mm),P<0.05].In addition,arthritis score in pristine-induced mice was 9.55±2.80 at 21 weeks after first injection with 0.5 ml pristine.H&E staining revealed a significant increase of synovial inflammation, cartilage and bone destruction after stimulated with pristine.Meanwhile,the expression levels of CD11b,CD11c,GR1,CD4,CD8 and CD154 were obviously increased in model-marking group when compared with that in control group.Conclusion: The pristine-induced model presents the similar histological and immunological distinctions with human rheumatism arthritis,which can mimic the pathogenesis of rheumatism arthritis.

0.05). Conclusions TVT is a safe, effective and minimally invasive surgical procedure for treatment of female stress urinary incontinence.

AIM: To investigate the changes of intracellular calcium ion (Ca2+) concentration in mouse H9c2 (2-1) cells transfected with or without FK506 binding protein 12.6(FKBP12.6) gene by ultrasound mediated destruction of microbubbles. METHODS: The pcDNA3.1-FKBP12.6 plasmid, mingled with albumin-coated microbubbles agents, was transfected into H9c2 (2-1) cells by ultrasound-mediated destruction of microbubbles. The H9c2 (2-1) cell growth state was investigated by inverted microscope. The changes of intracellular Ca2+ concentration was determined by laser scanning confocal microscope. The FKBP12.6 protein expression was checked by immunohistochemistry. RESULTS: As compared with control cells, the H9c2 (2-1) cells, transfected with FKBP12.6 gene, grew better, had higher gross intracellular Ca2+ concentration. CONCLUSION: FKBP12.6 gene augments Ca2+ concentration in mouse H9c2 (2-1) cells, enhances the contractibility of the myocardial cell, which may be helpful to improve the myocardial dysfunction.

Objective To discuss the efficacy of urinary continence in patients undergoing radical prostatectomy with bladder neck extension and mucosal eversion reconstruction anastomosis.Methods From August 2016 to November 2018,31 patients with prostate cancer underwent laparoscopic radical prostatectomy.The patients were 62-85 years old(mean 74.8 years),and the mean PSA score was 16.5 ng/ml(6.8-34.2 ng/ml).The pathological examination confirmed that the Gleason score was 6-9 and the prostate size was 44-83 ml.All patients underwent laparoscopic radical prostatectomy with bladder neck extension and mucosal eversion reconstruction anastomosis.Surgical procedure:After resection of the prostate in laparoscopic radical prostatectomy,the bladder neck was sutured at 6 o'lock position to narrow the bladder neck (" tennis racquet" reconstruction).The interval was 1 cm,and 2-3 needles were sutured,the distance between the neck of the bladder and the ureter was extended.The bladder neck mucosa and urethral mucosa eversion were performed.The posterior wall of the bladder neck was sutured at interval of 1 cm on both sides of the midline.After the knot was tightened,the posterior wall of the bladder was folded and bladder neck was elevated.The posterior wall of the bladder and the posterior wall of the urethra were sutured to reduce the distance between the bladder and the urethra.Finally,the bladder and urethra were anastomosed.The postoperative urinary continence recovery and the clinical effect were recorded.Results The operation time of 31 patients ranged from 80 to 210,with an average of 139.7 minutes.Intraoperative bleeding was 50-330 ml,with an average of 144.2 ml.None of the patients switched to open surgery during the operation,and there was no injury to large vessels and rectum,and no anastomotic leakage.Postoperative pathology showed 21 cases of pT2 stage,10 cases of pT3 stage,2 cases of positive margin,including 1 case of basal part and 1 case of apex part,both of which received medical castration therapy postoperatively.The surgical margin was positive in 2 cases (6.45％).31 patients removed the urinary catheter in ten days after surgery.17 cases (54.8％) recovered instantly urinary continence;7 cases (22.6％) urinary continence in 1 month after operation;4 cases (12.9％) urinary continence in 3 months after operation;and 3 cases (9.7％) urinary control in 6 months after operation.One case had urinary retention after removing the urinary catheter,and cathetered for 2 more weeks.After pulling out again,the urinary continence was good.Conclusions Laparoscopic radical prostatectomy with bladder neck extension and mucosal eversion reconstruction anastomosis may be helpful for early recovery of urinary continence.