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Objective@#To explore the molecular mechanism of miR-200b-3p regulates the proliferation, invasion, migration and apoptosis of pancreatic cancer cells by targeting vascular endothelial growth factor A (VEGFA).@*Methods@#The expression of miR-200b-3p in pancreatic cancer tissues and cell lines was detected by quantitative real-time fluorescence PCR (qRT-PCR). Pancreatic cancer PANC-1 cells were divided into NC group, miR-200b-3p mimic group, si-VEGFA group and si-VEGFA+ miR-200b-3p inhibitor group. The proliferation, migration and invasion of PANC-1 cells were measured by CCK-8 and Transwell assay. The apoptosis of PANC-1 cells was detected by Annexin V-FITC/PI double staining flow cytometry assay. The targeted relationship of miR-200b-3p and VEGFA was estimated by dual luciferase reporter gene assay and Western blotting.@*Results@#The expression of miR-200b-3p in pancreatic cancer tissues and cell lines was decreased. After miR-200b-3p was overexpressed in PANC-1 cells for 48 h, the cell viabilities of PANC-1 cells in NC group and miR-200b-3p mimic group were 1.250±0.028 and 0.983±0.044, the numbers of migrated cells were 402.700±21.530 and 158.000±17.620, the numbers of invaded cells were 478.300±31.050 and 170.000±32.470, and the cell apoptosis rates were (5.280±0.352)% and (7.430±0.393)%. The cell viability, migration and invasion of PANC-1 cells in miR-200b-3p mimic group were significantly decreased than those in NC group (t=5.060, P=0.007; t=8.796, P=0.001; t=6.863, P=0.002). The cell apoptosis rate in miR-200b-3p mimic group was significantly higher than that in NC group (t=4.076, P=0.015). The fluorescence intensity in VEGFA-WT group was 1.000±0.027, which was significantly higher than that in VEGFA-WT+ miR-200b-3p mimic group (0.632±0.048; t=6.637, P=0.003). The fluorescence intensities in VEGFA-MUT group and VEGFA-MUT + miR-200b-3p mimic group were 1.000±0.049 and 0.868±0.047, with no statistically significant difference (t=1.944, P=0.124). After miR-200b-3p was overexpressed in PANC-1 cells for 48 h, the expressions of VEGFA in NC group and miR-200b-3p mimic group were 1.000±0.058 and 0.762±0.020, respectively. The expression level in miR-200b-3p mimic group was lower than that in NC group (t=3.908, P=0.017). After transfection of PANC-1 cells with si-VEGFA or si-VEGFA + miR-200b-3p inhibitor for 48 h, the cell viabilities of PANC-1 cells in NC group, si-VEGFA group and si-VEGFA + miR-200b-3p inhibitor group were 1.300±0.058, 0.943±0.047 and 1.143±0.023, the numbers of migrated cells were 446.000±17.350, 206.300±19.360 and 428.300±30.330, and the numbers of invaded cells were 510.300±24.550, 175.700±24.290 and 473.700±35.530, with statistically significant differences (F=15.830, P=0.004, F=33.530, P=0.001, F=38.860, P<0.001). The cell viability, migration and invasion of PANC-1 cells in si-VEGFA group were significantly decreased than those in NC group (P=0.003, P<0.001, P<0.001). There was no significant difference between si-VEGFA + miR-200b-3p inhibitor group and NC group (P=0.107, P=0.854, P=0.671). The cell apoptosis rates in NC group, si-VEGFA group and si-VEGFA+ miR-200b-3p inhibitor group were (3.810±0.577)%, (7.373±0.482)% and (3.650±0.514)%, with a statistically significant difference (F=16.020, P=0.004). The cell apoptosis rate in si-VEGFA group was significantly higher than that in NC group (P=0.007), but there was no significantly difference between si-VEGFA + miR-200b-3p inhibitor group and NC group (P=0.975).@*Conclusion@#miR-200b-3p suppresses the proliferation, invasion and migration and promotes the apoptosis of pancreatic cells by down-regulating VEGFA.
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Objective To explore the molecular mechanism of miR-200b-3p regulates the prolifera-tion,invasion,migration and apoptosis of pancreatic cancer cells by targeting vascular endothelial growth factor A (VEGFA). Methods The expression of miR-200b-3p in pancreatic cancer tissues and cell lines was detec-ted by quantitative real-time fluorescence PCR (qRT-PCR). Pancreatic cancer PANC-1 cells were divided into NC group,miR-200b-3p mimic group,si-VEGFA group and si-VEGFA + miR-200b-3p inhibitor group. The proliferation,migration and invasion of PANC-1 cells were measured by CCK-8 and Transwell assay. The apop-tosis of PANC-1 cells was detected by Annexin V-FITC/ PI double staining flow cytometry assay. The targeted relationship of miR-200b-3p and VEGFA was estimated by dual luciferase reporter gene assay and Western blotting. Results The expression of miR-200b-3p in pancreatic cancer tissues and cell lines was decreased. After miR-200b-3p was overexpressed in PANC-1 cells for 48 h,the cell viabilities of PANC-1 cells in NC group and miR-200b-3p mimic group were 1. 250 ± 0. 028 and 0. 983 ± 0. 044,the numbers of migrated cells were 402. 700 ± 21. 530 and 158. 000 ± 17. 620,the numbers of invaded cells were 478. 300 ± 31. 050 and 170. 000 ± 32. 470,and the cell apoptosis rates were (5. 280 ± 0. 352)％ and (7. 430 ± 0. 393)％ . The cell viability,migration and invasion of PANC-1 cells in miR-200b-3p mimic group were significantly decreased than those in NC group (t = 5. 060,P = 0. 007;t = 8. 796,P = 0. 001;t = 6. 863,P = 0. 002). The cell apop-tosis rate in miR-200b-3p mimic group was significantly higher than that in NC group (t = 4. 076,P = 0. 015). The fluorescence intensity in VEGFA-WT group was 1. 000 ± 0. 027,which was significantly higher than that in VEGFA-WT + miR-200b-3p mimic group (0. 632 ± 0. 048;t = 6. 637,P = 0. 003). The fluorescence intensi-ties in VEGFA-MUT group and VEGFA-MUT + miR-200b-3p mimic group were 1. 000 ± 0. 049 and 0. 868 ± 0. 047,with no statistically significant difference (t = 1. 944,P = 0. 124). After miR-200b-3p was overex-pressed in PANC-1 cells for 48 h,the expressions of VEGFA in NC group and miR-200b-3p mimic group were 1. 000 ± 0. 058 and 0. 762 ± 0. 020,respectively. The expression level in miR-200b-3p mimic group was lower than that in NC group (t = 3. 908,P = 0. 017). After transfection of PANC-1 cells with si-VEGFA or si-VEGFA + miR-200b-3p inhibitor for 48 h,the cell viabilities of PANC-1 cells in NC group,si-VEGFA group and si-VEGFA + miR-200b-3p inhibitor group were 1. 300 ± 0. 058,0. 943 ± 0. 047 and 1. 143 ± 0. 023,the numbers of migrated cells were 446. 000 ± 17. 350,206. 300 ± 19. 360 and 428. 300 ± 30. 330,and the numbers of invaded cells were 510. 300 ± 24. 550,175. 700 ± 24. 290 and 473. 700 ± 35. 530,with statisti-cally significant differences (F = 15. 830,P = 0. 004,F = 33. 530,P = 0. 001,F = 38. 860,P < 0. 001). The cell viability,migration and invasion of PANC-1 cells in si-VEGFA group were significantly decreased than those in NC group (P = 0. 003,P < 0. 001,P < 0. 001). There was no significant difference between si-VEGFA + miR-200b-3p inhibitor group and NC group (P = 0. 107,P = 0. 854,P = 0. 671). The cell apop-tosis rates in NC group,si-VEGFA group and si-VEGFA + miR-200b-3p inhibitor group were (3. 810 ± 0. 577)％,(7. 373 ± 0. 482)％ and (3. 650 ± 0. 514)％,with a statistically significant difference (F =16. 020,P = 0. 004). The cell apoptosis rate in si-VEGFA group was significantly higher than that in NC group (P = 0. 007),but there was no significantly difference between si-VEGFA + miR-200b-3p inhibitor group and NC group (P = 0. 975). Conclusion miR-200b-3p suppresses the proliferation,invasion and migration and promotes the apoptosis of pancreatic cells by down-regulating VEGFA.
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To date, the efficacy of radical surgery (RS) versus conservative surgery (CS) for liver hydatid cysts (LHC) remains controversial. This meta-analysis was conducted to compare the two interventions. PubMed, Embase, and Web of Science were searched from their inceptions until June 2016. Meta-analysis was performed using STATA 12.0 software. We identified 19 eligible studies from 10 countries by retrieval. In total, 1853 LHC patients who received RS were compared with 2274 patients treated by CS. The risk of postoperative overall complication, biliary fistula, and recurrence was significantly lower, and operation time was significantly longer in the RS group. However, no statistically significant differences were found in terms of mortality risk and the duration of hospital stay between RS and CS. No significant publication biases were observed in all the above analyses. In conclusion, RS reduces the rates of postoperative complications and recurrence, whereas no trend toward such a reduction in mortality was observed in LHC patients.
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Humans , Echinococcosis, Hepatic , Mortality , General Surgery , Length of Stay , Operative Time , Postoperative Complications , Epidemiology , Recurrence , Treatment OutcomeABSTRACT
Objective To observe the safety and clinical efficacy of enhanced recovery after surgery in the perioperation period of hepatectomy.Methods 92 patients with primary hepatic cancer who underwent surgical resection in our hospital from December 2014 to May 2016 were divided into two groups,50 patients received well-organized and consecutive clinical interventions guided by ERAS.42 patients underwent traditional perioperative management.Results Compared with traditional group,ERAS group had reduced hospital cost [(40 633.12 ± 6 336.46) RMB vs.(46 139.23 ± 9 605.88) RMB,P ＜ 0.05],shorter postoperative hospital stay[(10.24 ± 1.6) d vs.(13.35 ± 4.86) d,P ＜ 0.05],earlier flatus and defecation [(33.34 ±6.01) h vs.(50.31 ± 3.53) h,P ＜ 0.05],and improved satisfaction rate for pain management[45/50(90％) vs.22/42(52.4％),P ＜ 0.05];while the postoperative adverse events and complications of the two groups showed no difference (P ＞ 0.05).Conclusion The application of ERAS in the perioperation period of hepatectomy is safe and effective,reducing hospital cost,postoperative hospital stay,improving satisfaction rate to pain management,and facilitating recovery in hepatic surgery.
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Objective To compare the therapeutic efficacy of combined biliary stent and 125I seed intracavity irradiation with palliative surgery in the treatment of extrahepatic ductal cholangiocarcinoma.Methods A prospective analysis was conducted on 142 patients with cholangiocarcinoma who were treated in The First Affiliated Hospital of Bengbu Medical College from January 2012 to December 2015.There were 80 patients who underwent percutaneous biliary metal stenting combined with 125I particles implantation (the stenting-particle group) and 62 patients who were treated by palliative biliary drainage (the surgical group).The surgical group included R1 resection in 17 patients,R2 resection in 26 patients and biliary enteric drainage in 19 patients).The levels of jaundice,liver function,survival time,hospitalization time and hospitalization cost before and after therapy were analyzed.Results Jaundice was effectively alleviated in the two groups after a short period.The liver function in the 2 groups improved significantly at 1,3 and 6 months when compared with that before operation,(P ＜ 0.05).The average hospitalization time of the stenting-particle group and the surgery group were (16.5 ± 5.0) days and (25.5 ± 10.5) days,respectively,(P ＜ 0.01).The average hospitalization cost of the stenting-particle group and the surgery group were (39 622.0 ± 7 666.4) yuan and (59 562.0 ± 24 218.2) yuan,respectively,(P ＜ 0.05).The average survival time of the stenting-particle group and the surgery group were (12.2 ± 5.1) months and (12.69 ± 7.46) months,respectively,and the difference was not significantly different (P ＞ 0.05).Conclusions For patients with extrahepatic ductal cholangiocarcinoma who were not suitable for radical surgery,percutaneous biliary stenting combined with 125I seed brachytherapy effectively reduced jaundice,improved liver function,shortened average length of hospital stay and reduced average cost of hospitalization.When compared with palliative surgery,it was a minimally invasive,easy,safe and efficacious treatment,especially for elderly patients with poor physical conditions.
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Objective To explore the efficacy difference of biliary stent combined with 125I seed intracavity irradiation and palliative biliary drainage in the treatment of obstructive jaundice caused by pancreatic head cancer. Methods The clinic date of 95 patients with pancreatic head cancer who underwent palliative surgery from June 2010 to June 2015 were analyzed retrospectively. 46 of the cases were treated with percutaneous biliary metal stent implantation combined with 125I seed intracavity irradiation (stent group), and the other 49 cases were treated with palliative biliary enterostomy (surgery group). The levels of jaundice, liver function, tumor size, survival time and hospitalization cost were compared between the patients before and after treatment. Paired t-test was applied for statistical analysis. Results Both the two treatment coulel effectively relieve jaundice and improve liver function. The mean TBIL of surgery group and stent group was (29.4±9.6) and (21.0±8.0)μmol/L after three months treanment, (40.3±11.0) and (24.4±9.6)μmol/L after six months treatment, respectively. Tumor maximum diameter of the two groups were (37.9 ± 4.5) mm and (33.5 ± 6.0)mm after three months treatment, (45.9 ± 5.0) mm and (37.0 ± 6.5) mm after six months, respectively. Each time point between them had statistical significance (t=-4.235,-5.476,-3.654,-6.702, P<0.05). The mean survival time was (12.83 ± 0.80) months in the stent group and (9.06±0.49) months in the surgery group, the difference was statistically significant (t=-3.49, P<0.01). The mean hospitalization cost of the stent group was (38 453.0 ± 7 282.0) yuan and the surgery group was (43 057.0 ± 7 667.4) yuan, the difference was statistically significant (t= 2.759, P<0.05). Conclusion For inoperable pancreatic head cancer patients, compared with palliative biliary enteric drainage, percutaneous biliary stent placement combined with 125I seed intracavity irradiation can effectively relieve biliary obstruction, improve liver function, inhibit tumor growth and prolong survival time.
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<p><b>OBJECTIVE</b>To investigate the effect of MSX2 interference on epithelial-mesenchymal transitions (EMT) of pancreatic cancer cell line PANC-1.</p><p><b>METHODS</b>Three vectors containing short hairpin RNAs (shRNAs) of MSX2 (shMSX2-1, shMSX2-2, and shMSX2-3) and the empty vector (negative control) were transfected separately into PANC-1 cell line with Lipofectamine2000. Real-time RT-PCR and Western blotting were used to observe changes in the expressions of MSX2, E-cadherin, and vimentin in the cells. CCK-8 assay was used to assess the changes in the cell growth, and wound scratch assay and Transwell assay were employed to evaluate the cell invasion and metastasis after the transfection.</p><p><b>RESULTS</b>Among the 3 shRNA, shMSX2-1 showed the highest interference efficiency. MSX2 knockdown by the specific shRNA of MSX2 significantly increased E-cadherin expressions, lowered vimentin expressions, and suppressed the invasion, metastasis and proliferation of the cells (P<0.05). MSX2 knockdown also resulted in morphological changes of the cells into cobblestone-like cells in close contact. RT-PCR results revealed significantly reduced mRNA expressions of the transcription factors snail and twist (P<0.05) without affecting slug and zeb1 expressions in the cells with MSX2 knockdown. Conclusion MSX2 knockdown can reverse EMT and induce MET in PANC1 cells, in which process the transcription factors snail and twist may play a role.</p>
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Humans , Cadherins , Metabolism , Cell Line, Tumor , Cell Proliferation , Epithelial-Mesenchymal Transition , Homeodomain Proteins , Metabolism , Nuclear Proteins , Metabolism , Pancreas , Pancreatic Neoplasms , Pathology , RNA, Small Interfering , Snail Family Transcription Factors , Transcription Factors , Metabolism , Transfection , Twist-Related Protein 1 , Metabolism , Vimentin , MetabolismABSTRACT
Objective To evaluate the clinical application of the treatment on malignant obstructive jaundice by combination of percutaneous biliary stent placement and 125 I particles intraluminal implanation.Methods As a prospective study , 32 patients with malignant obstructive jaundice who either have no opportunity for radical operations or unwilling to be surgically treated were enrolled to be treated with percutaneous biliary stent placement and 125 I particles intraluminal implanation.Biochemical routine tests , blood routine tests , tumor markers , coagulation function , color ultrasound , CT and magnetic resonance cholangiopancreatography were conducted prior to the operation to obtain general information of the clinical status of the patients and the tumor and the site of obstruction.Percutaneous transhepatic cholangial drainage was performed first under the B -type ultrasound system.After one week , biliary stents were placed under DSA.According to the stent expansion presented by real-time DSA imaging , 125 I particles were implanted simultaneously or afterwards.Routine biochemical tests and cholangiopancreatography under DSA were conducted in one week , one month and three months after the implantation.Variance analysis was performed with repeated measurements to compare the difference of liver function indexes pre -and post-operation.Meanwhile, 125I particle displacement, falling off and stent patency were observed.After three months, the tumor size was measured by CT.Student t-test was used to compare the tumor sizes of pre-and post-operation.Results The symptoms of jaundice resolved and the general physical conditions improved in 32 patients substantially.The total bilirubin level decreased from preoperative level of (302 ±169)μmol/l to the level of (108 ±50)μmol/l at one week following the surgery , and the indirect bilirubin level decreased from preoperative level of ( 250 ±160 )μmol/l to the level of ( 85 ±43 )μmol/l at one week following the surgery(F=76.32,58.23,P<0.01).The maximal diameters of the tumors decreased from preoperative size of (3.78 ±1.14)cm to the size of (3.14 ±1.28)cm at three months following the surgery, and the minimal diameters of the tumors decreased from preoperative size of ( 2.80 ±0.88 ) cm to the size of ( 1.93 ± 1.00)cm at three months following the surgery, and the differences were statistically significant (t=5.11, 6.73,P<0.05).By using Kaplan-Meier survival curve, the average survival periods were ( 9.9 ± 0.7) months.Conclusions Percutaneous biliary stent placement and 125 I particles intraluminal implanation have definite short-term effects in prolonging survival time , stent patency time and improving the living standard of the patients.The technique is safe and simple.It only needs small incision , has no absolute contraindications and can be applied repeatedly .
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BACKGROUND:Hepatocyte transplantation can improve biochemical parameters and survival of animals with acute liver failure.However,cell sources.immunological rejection,distribution and histomorphological alternation of transplanted hepatocytes are important issues for its wide application in clinic.OBJECTIVE:To observe early-stage histomorphological and ultrastructural changes of SV40LT antigen gene transfected hepatocytes after transplanted into rat spleen.DESIGN,TIME AND SETTING:Randomized controlled animal trial was performed at Laboratory of National Hepatobiliary and Enteric Surgery Research Center.Ministry of Health from March to December 2001.MATERIALS:Sixty male Wistar rats were selected for hepatocyte transplantation.METHODS:Sixty rats were randomly divided into 4 groups(n=15).Primary cell group and primary cell plus cyclosporine A group were intrasplenically transplanted with primarily cultured hepatocytes;SV40LT antigen gene group and SV40LT antigen gene plus cyclosporine A group were intrasplenic transplanted with SV40LT antigen gene transfected hepatocytes.Twenty-four hours before and 14 days after transplantation,the rats in primary cell group and SV40LT antigen gene group were injected with 0.5 mL normal saline through tail vein daily,while the other groups were injected with cyclosporine A (10mg/kg per day).MAIN OUTCOME MEASURES:The spleen of one rat was harvested every day postoperatively for light microscopic and electron microscopic examinations to observe survival rate,histomorphological and ultrastructural features of transplanted hepatocytes for 14 days.RESULTS:Compared with primary cell group and SV40LT antigen gene group,the survival rate of transplanted hepatocytes in the other groups was significantly increased(P<0.05),but the histomorphological and ultrastructural changes were minor.There were no significant differences in the survival rate between SV40LT zntigen gene transfected hepatocytes and primarily cultured hepatocytes during the first week after transplantation(P>0.05),while the survival rate in SV40LT antigen gene transfected hepatocytes was significantly higher than primarily cultured hepatocytes 8-14 days postoperatively(P<0.01).CONCLUSION:Under treatment of cyclosporine A,the SV40LT antigen gene transfected hepatocytes can maintain a stable ultrastructure and a long survival rate.
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Objective To investigate the clinical value of improved percutaneous tramhepatic cholangiographic drainage(PTCD)and percutaneous placement of biliary metallic stents on the treatment of malignant obstructive jaundice.Methods Thirty three patients with malignant biliary obstruction were treated by the improved PTCD guided by ultrasound followed by pereutaneous placement of biliary tract metallic stcnts 1～3 weeks later.Results Improved PTCD WaS succeeded in all the patients,30 patients(90.9%)accepted percutaneous placement of biliary metallic stents successfully,3 patients who failed in stents placement accepted PTCD again for palliative management of iallndice.Thofle who failed in stent placement were found to have bleeding from the PTCD tube,but recovered several days later.Jaundice alleviated in all the patients.The patients were followed up for 1～29 months,medium follow-up period is 14.8 months.Six cases were found lever and jaundice,among that,two ageepted improved PTCD again,one recovered after antibioic treatment,the remaining 3 patients failed in further treatment due to the metastasis of the tumor.Condusion Pereutaneous placement of biliary metallic stents after improved PTCD is a safe and effeetive method to relieve malignant biliary obstruction,it can improve life quality,prolong survival time.This method could be a good choice for treating the patients with mnlignant biliary obstruction without operation.
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BACKGROUND: Hepatocyte transplantation can improve biochemical parameters and survival of animals with acute liver failure. However, cell sources, immunological rejection, distribution and histomorphological alternation of transplanted hepatocytes are important issues for its wide application in clinic. OBJECTIVE: To observe early-stage histomorphological and ultrastructural changes of SV40LT antigen gene transfected hepatocytes after transplanted into rat spleen. DESIGN, TIME AND SETTING: Randomized controlled animal trial was performed at Laboratory of National Hepatobiliary and Enteric Surgery Research Center, Ministry of Health from March to December 2001. MATERIALS: Sixty male Wistar rats were selected for hepatocyte transplantation. METHODS: Sixty rats were randomly divided into 4 groups (n=15). Primary cell group and primary cell plus cyclosporine A group were intrasplenically transplanted with primarily cultured hepatocytes; SV40LT antigen gene group and SV40LT antigen gene plus cyclosporine A group were intrasplenic transplanted with SV40LT antigen gene transfected hepatocytes. Twenty-four hours before and 14 days after transplantation, the rats in primary cell group and SV40LTor antigen gene group were injected with 0.5 mL normal saline through tail vein daily, while the other groups were injected with cyclosporine A (10 mg/kg per day). MAIN OUTCOME MEASURES: The spleen of one rat was harvested every day postoperatively for light microscopic and electron microscopic examinations to observe survival rate, histomorphological and ultrastructural features of transplanted hepatocytes for 14 days. RESULTS: Compared with primary cell group and SV40LT antigen gene group, the survival rate of transplanted hepatocytes in the other groups was significantly increased (P 0.05), while the survival rate in SV40LT antigen gene transfected hepatocytes was significantly higher than primarily cultured hepatocytes 8-14 days postoperatively (P
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Objective To summarize the experience of diagnosis and treatment of solid-pseudopapillary tumor of pancreas (SPTP).Methods The clinical data of 15 patients with SPTP who were treated in our hospital from March,2003 to March,2006 were analyzed retrospectively.Results The 15 cases were all women, and the average age was 29.4 years.The chief manifestations were abdominal mass, abdominal pain or other abdominal discomfort. None of the 15 cases had history of pancreatitis or abdominal trauma, and no long history of drinking or smoking. Six cases were negative for CEA,CA50,CA199,CA125 and other tumor markers.Solid and solid-cystic masses in pancreas or solid and solid-cystic tumors in retroperitoneum were found both by B-mode ultrasonography and CT examinations. Preoperative fasting blood sugar was within normal limits. The tumor in 8 cases was located in the pancreatic head, in 6 cases was in the body and tail of pancreas, and in 1 case was in the neck of pancreas. The diameter of the tumors was 2.5-10 cm. No metastasis was found in the abdominal cavity or liver. Local excision was performed in 6 cases, distal pancreatectomy was performed in 5 cases, including 2 cases combined with splenectomy, and pancreaticoduodenectomy was performed in 3 cases, segmental pancreatectomy was performed in one patient with tumor in the neck of pancreas. The 15 cases showed typical pathologic manifestation of SPTP by microscopy. At followed up for 16-52 months, no evidence of recurrence or metastasis in these cases was found.Conclusions Solid-pseudopapillary tumor of pancreas primarily affects young women, and it may be located in any part of pancreas. Surgical resection is recommended as the treatment of choice, and the prognosis is good.