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Objective:To investigate the effects of orlistat on the viability of human gall-bladder cancer (GBC) cells.Methods:The experimental study was conducted. The human GBC NOZ cells with high expression of FSAN was screened out through in vitro cultivating human GBC-SD, SGC-996 and NOZ cells. The cell proliferation assay, clone formation assay and protein detection experiment were used to analysis of the effects of orlistat on the viability of human GBC cells. Cell grouping: NOZ cells cultured with medium were set as the control group, cultured with medium + 10 μmol/L orlistat were set as the low-dose orlistat group, cultured with medium + 100 μmol/L orlistat were set as the high-dose orlistat group, respectively. Observation indicators: (1) expression of FASN protein in human GBC cells; (2) effects of orlistat on the proliferation of human GBC NOZ cells; (3) effects of orlistat on apoptosis of human GBC NOZ cells. Measurement data with normal distribution were represented as Mean± SD, the ANOVA test was used for comparison between groups and the least significant difference method was used for pairwise comparison. Results:(1) Expression of FASN protein in human GBC cells. Results of western blot showed that the relative expression of FASN protein in human GBC NOZ, GBC-SD and SGC-996 cells was 0.57±0.06, 0.12±0.04 and 0.10±0.02, respectively, showing a significant difference among them ( F=115.67, P<0.05). There were significant differences between the NOZ cells and the GBC-SD or the SGC-996 cells ( P<0.05), and there was no significant difference between the GBC-SD cells and the SGC-996 cells ( P>0.05). (2) Effects of orlistat on the proliferation of human GBC NOZ cells. ① Results of cell proliferation assay showed that the absorbance value of NOZ cells was 2.34±0.12, 1.57±0.08 and 1.07±0.13 in the control group, low-dose orlistat group and high-dose orlistat group, respectively, showing a significant difference among them ( F=205.88, P<0.05). ② Results of clone formation assay showed that the number of NOZ cells clones was 257±23, 153±11 and 83±11 in the control group, low-dose orlistat group and high-dose orlistat group, respectively, showing a significant difference among them ( F=92.64, P<0.05). ③Results of western blot showed that the relative expression of Cyclin-D1 protein of NOZ cells was 2.31±0.10, 1.52±0.05 and 1.23±0.11 in the control group, low-dose orlistat group and high-dose orlistat group, respectively, showing a significant difference among them ( F=120.73, P<0.05). The relative expression of CDK-4 protein of NOZ cells was 1.58±0.04, 1.21±0.02 and 1.19±0.04 in the control group, low-dose orlistat group and high-dose orlistat group, respectively, showing a signifi-cant difference among them ( F=110.45, P<0.05). (3) Effects of orlistat on apoptosis of human GBC NOZ cells. Results of western blot showed that the relative expression of Bcl-2 protein of NOZ cells was 1.07±0.03, 0.36±0.03 and 0.15±0.02 in the control group, low-dose orlistat group and high-dose orlistat group, respectively, showing a significant difference among them ( F=1 242.93, P<0.05). The relative expression of Bax protein of NOZ cells was 0.51±0.03, 0.38±0.05 and 1.38±0.04 in the control group, low-dose orlistat group and high-dose orlistat group, respectively, showing a signifi-cant difference among them ( F=583.51, P<0.05). Conclusion:Orlistat can inhibit the growth of human GBC NOZ cells and promote their apoptosis.
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Objective:To study the expression of forkhead box P1 (FOXP1) in intrahepatic cholangiocarcinoma (ICC), and its clinicopathological and prognostic significance.Methods:The clinical data of ICC patients treated with radical resection at Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine from January 1, 2013 to December 12, 2019 were retrospectively analysed. Of 48 ICC patients, there were 24 males and 24 females, with age of (59.1±10.1) years old (range 42 to 83 years old). Their clinicopathological data, including age, gender, tumor size, degree of differentiation, and staging were recorded. Immunohistochemical method was used to detect the expression of FOXP1 protein in ICC cancer tissues and the corresponding adjacent normal tissues. Kaplan-Meier method was used to calculate survival rates and to construct survival curves of patients. Cox regression model was used to analyze factors affecting prognosis of patients.Results:Forty-eight ICC cancer tissues and 40 corresponding paracancerous tissues were collected. The positive rates of FOXP1 proteins in ICC were significantly lower than the adjacent normal tissues [54.2%(26/48) vs. 92.5%(37/40), χ 2=15.76, P<0.05]. The degrees of tumor differentiation, lymph node metastasis, organ invasion and TNM staging were related to expression of FOXP1 ( P<0.05). Forty-two patients were followed-up with a median follow-up time of 11.5 (7.75, 19.25) months. Cox multivariate analysis revealed that invasion to adjacent organs, lymph node metastasis, high TNM staging (stage Ⅲ) and negative expression of FOXP1 were independent risk factors affecting overall survival of ICC patients. The overall survival and recurrence-free survival of FOXP1-positive ICC patients were 17.5 months and 15.5 months, which were significantly higher than the 14.0 months and 11.1 months, respectively, in FOXP1-negative patients. Conclusion:Negative FOXP1 expression was closely correlated with aggressive biological behavior and poor prognosis of ICC. FOXP1 may be used as new diagnostic and therapeutic targets.
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Laparoscopic cholecystectomy (LC) is the first preferred treatment of benign gallbladder diseases such as gallbladder stones and gallbladder polyps, however bile duct injury is a serious complication of LC. Although bile duct injury is a rare complication, improper treatments will seriously affect the quality of life or even threaten life. Therefore, the prevention and correct treatments of bile duct injury in LC are crucial. Based on domestic and overseas researches, the authors investigate risk factors for bile duct injury in LC, share experiences of timely detection, diagnosis and treatment, so as to provide references for hepatic and biliary surgeons.
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Objective To investigate application value of cutter stapler in transumbilical single port laparoscopic left lateral lobectomy.Methods The retrospective cohort study was adopted.The clinical data of 26 patients who underwent transumbilical single port laparoscopic left lateral lobectomy at the Shengjing Hospital of China Medical University from January 2010 to February 2016 were collected.Nine patients who received liver parenchyma using ultrasonic knife were allocated into the ultrasonic knife group,17 patients who received liver parenchyma using cutter stapler were allocated into the cutter stapler group.Observation indicators included (1) operation situations:operation time,volume of intraoperative blood loss,postoperative complications,time of postoperative bowel function recovery,time of abdominal cavity drainage tube removal,duration of postoperative hospital stay.(2) Postoperative reexamination and follow-up:ultrasound or computed tomography (CT) examination was performed when necessary for detecting local exudation or encapsulated effusion.The patients were followed up at postoperative 1 to 3 months with telephone interview for whether with abdominal distension or abdominal pain till March 2016.Measurement data with normal distribution were presented as (x) ± s and analyzed by using t test.Measurement data with skewed distribution were presented as M (range) and analyzed by ranksum text.Comparison of count data was analyzed by the Fisher' s exact probility.Results (1) Operation situations:all the 26 patients received transumbilical single port laparoscopic left lateral lobectomy with no conversion to porous laparoscopic surgery or open surgery.The operation time was (114 ± 54) minutes,the volume of intraoperative blood loss was 100 mL (range,20-800 mL),and no intraoperative blood transfusion was adopted.The operation time and volume of intraoperative blood loss were (135 ±43)minutes and 200 mL (range,20-800 mL) in the ultrasonic knife group,(103 ±57)minutes and 100 mL (range,20-300 mL) in the cutter stapler group,respectively,showing no statistically significant difference between the 2 groups (t =1.500,Z =-0.961,P ＞ 0.05).All the 26 patients recovered well after surgery,with no postoperative complications as postoperative hemorrhage,bile leakage,incision infection or death.The time of postoperative bowel function recovery,time of abdominal cavity drainage tube removal and duration of postoperative hospital stay was (1.5 ±0.4) days,(5.8 ± 2.0) days and (7.0 2.0) days in the ultrasonic knife group,(1.1 ± 0.3) days,(4.1 ±1.1) days and (4.9 ± 1.4) days in the cutter stapler group,respectively,showing statistically significant differences between the 2 groups (t =2.599,2.875,3.036,P ＜ 0.05).(2) Postoperative reexamination and follow-up:of 26 patients,11 patients received ultrasound or CT examination after surgery and detected no obvious local exudation or encapsulated effusion,with no special treatment.The other 15 patients didn't receive ultrasound or CT examination.All the 26 patients were followed up for 1-3 months,with no occurrence of upper abdominal distension or abdominal pain.Conclusion Transumbilical single port laparoscopic left lateral lobectomy is safe and feasible,the application of cutter stapler is helpful to safety and success of the operation,further accelerating the postoperative recovery of patients.
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Objective To establish immortalized human hepatocyte lines for studies of bioartificial liver,hepatocyte transplantation,and drug metabolism in vitro.Methods Primary human hepatocytes were isolated by 4-step perfusion technique with collagenase and transfected with recombinant retrovirus containing Simian virus 40 large T antigen(SV40 LT).Subsequently,immortalized human hepatocytes were evaluated by analysis of gene expression and functional characteristics in vitro.Results Two immortalized human hepatocyte lines,HepLi2 and HepLi3,were obtained after primary human hepatocytes being infected by SV40 LT containing recombinant retrovirus for 3-4 weeks.The immortalized human hepatocytes showed classical appearance of hepatocyte observed by phase contrast microscope.The protein expression of SV40 LT in HepLi2 and HepLi3 cells were detected by Western blotting.The mRNA expressions of albumin(Alb),glutathione S-transferase(GST-p),human blood coagulation factor X(HBCF-X)and β-actin in HepLi2 and HepLi3 cells were detected by reverse transcription polymerase chain reaction(RT-PCR),and the mRNA expressions of cytochrome(CY)450 subtypes(CYP3A5,CYP2E1,CYP2C8-19 and CYP3A4)in HepLi2 and HepLi3 cells were also observed by RT-PCR.Levels of alanine transaminase (ALT),lactate dehydrogenase(LDH)and Alb were detected in the supernatant of immortalized human hepatoeyte culture.Conclusions The immortalized human hepatocyte lines have the biological characteristics of primary human hepatocytes and have the CYP450 functions of hepatocytes,which may be heIDful for the studies of bioartificial liver,heoatocvte transplantation and drug metabolism in vitro.