Antiproliferative effects of cinobufacini on human hepatocellular carcinoma HepG2 cells detected by atomic force microscopy

AIM:To investigate the antiproliferative activity of cinobufacini on human hepatocellular carcinoma HepG2 cells and the possible mechanism of its action.METHODS:HepG2 cells were treated with different concentrations of cinobufacini.Cell viability was measured by methylthiazolyl tetrazolium(MTT) assay.Cell cycledistribution was analyzed by flow cytometry(FCM).Cytoskeletal and nuclear alterations were observed by fluorescein isothiocyanate-phalloidin and DAPI staining under a laser scanning confocal microscope.Changes in morphology and ultrastructure of cells were detected by atomic force microscopy(AFM) at the nanoscale level.RESULTS:MTT assay indicated that cinobufacini significantly inhibited the viability of HepG2 cells in a dosedependent manner.With the concentration of cinobufacini increasing from 0 to 0.10 mg/m L,the cell viability decreased from 74.9% ± 2.7% to 49.41% ± 2.2% and 39.24% ± 2.1%(P < 0.05).FCM analysis demonstrated cell cycle arrest at S phase induced by cinobufacini.The immunofluorescence studies of cytoskeletal and nuclear morphology showed that after cinobufacini treatment,the regular reorganization of actin filaments in HepG2 cells become chaotic,while the nuclei were not damaged seriously.Additionally,high-resolution AFM imaging revealed that cell morphology and ultrastructure changed a lot after treatment with cinobufacini.It appeared as significant shrinkage and deep pores in the cell membrane,with larger particles and a rougher cell surface.CONCLUSION:Cinobufacini inhibits the viability of HepG2 cells via cytoskeletal destruction and cell membrane toxicity.

Laparoscopic management of gastric gastrointestinal stromal tumors: A retrospective 10-year single-center experience

AIM To determine the feasibility,safety,and oncological outcome of laparoscopic resection of gastric gastrointestinal stromal tumors(GISTs)based on favorable or unfavorable location.METHODS Our hospital database included 207 patients who underwent laparoscopic removal of gastric GISTs from January 2004 to September 2015.Patient demographics,clinical presentation,surgery,histopathology,postoperative course,and oncological outcomes were reviewed and analyzed.RESULTS Gastric GIST in favorable locations was present in81/207(39.1%)cases,and in unfavorable locations in 126/207(60.9%)cases.Overall mean tumor size was 3.28±1.82 cm.No conversions occurred,and complete R0 resection was achieved in 207(100%)cases.There were three incidences of iatrogenic tumor rupture.The feasibility and safety of laparoscopic surgery were comparable in both groups with no statistical difference between unfavorable and favorable location groups,respectively:for operative time:83.86±44.41 vs 80.77±36.46 min,P=0.627;conversion rate:0%vs 0%;estimated blood loss:27.74±45.2vs 29.59±41.18 m L,P=0.780;tumor rupture during surgery:0.90%vs 2.82%,P=0.322;or postoperative complications:3.74%vs 7.04%,P=0.325.The follow-up period recurrence rate was 1.89%with no significant differences between the two groups(3.03%vs 0%,P=0.447).Overall 5-year survival rate was98.76%and survival rates were similar between the two groups:98.99%vs 98.39%,P=0.623(unfavorable vs favorable,respectively).CONCLUSION The laparoscopic approach for gastric GISTs is safe and feasible with well-accepted oncological surgical outcomes.Strategies for laparoscopic resection should be selected according to the location and size of the tumor.Laparoscopic treatment of gastric GISTs in unfavorable locations should not be restricted in gastrointestinal centers.

Laparoscopy-assisted total gastrectomy with trans-orally inserted anvil(OrVil^(TM)):A single institution experience

AIM:To investigate the feasibility of laparoscopyassisted total gastrectomy(LATG)using trans-orally inserted anvil(OrVilTM)in terms of operative characteristics and short term outcomes. RESULTS:Characteristics of 27 patients with gastric cancer who underwent LATG from October 2009 to October 2012 in the Foshan Affiliated Hospital of South Medical University were retrospectively reviewed. Among these patients,six were reconstructed by minilaparotomy and 21 by OrVilTM.The clinicopathological characteristics,total operation time,total blood loss, abdominal incision and complications of anastomosis including stenosis and leakage,were compared between the groups undergoing LATG with OrVilTM and the group undergoing minilaparotomy. RESULTS:The operations were successfully performed on all the patients without intraoperative complications or conversion to open surgery.Two(10%)patients received palliative procedure under laparoscope who were prepared for LATG preoperatively.One case had hepatic metastatic carcinoma and 1 case had tumor recurrence near the anastomosis 8 mo after surgery.The mean follow-up duration was 10 mo(range,2-24 mo). Operation time was significantly reduced by the use of OrVilTM(198.42±30.28 min vs 240.83±8.23 min). The postoperative course with regard to occurrence of stenosis and leakage was not different between the two groups.There were no significant differences in estimated blood loss.The upper abdominal incision was smaller in OrVilTM group than in minilaparotomy group (4.31±0.45 cm vs 6.43±0.38 cm). CONCLUSION:LATG using OrVil TM is a technically feasible surgical procedure with sufficient lymph node dissection,less operation time and acceptable morbidity.