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Objective@#To study the expression of Wnt2b protein and the epithelial-mesenchymal transition related markers in tissues of carcinoma of bile duct and normal bile duct to determine the clinical significance. The relationships between the expression levels and clinicopathological characteristics were analyzed, and the correlation between Wnt2b and epithelial interstitial transformation (EMT), tumor invasion and metastasis were studied.@*Methods@#A total of 60 patients with cholangiocarcinoma and 30 patients with normal bile duct tissues admitted to the Affiliated Hospital of Qingdao University from December 2008 to December 2013 were studied. The expressions of Wnt2b, E-cadherin and Vimentin protein were detected by SP immunohistochemical staining. The patients were classified according to the expressions of these proteins. Analyses were conducted on the relationships of these proteins with clinical characteristics of the patients with cholangiocarcinoma.@*Results@#The positive expression rate of Wnt2b protein in carcinoma of bile duct tissues was 90.0%, which was significantly higher than that in normal bile duct tissues (χ2=38.1, P<0.05). The positive expression rate of Wnt2b in T3 + T4 was significantly higher than that in T1 + T2 (P<0.05). There were no correlations of the expression with patients’ age, gender, tumor location and degree of tumor differentiation (P>0.05). Patients with lymph node metastasis had a significantly higher positive expression of Vimentin than patients with no lymph node metastasis, and the loss of E-cadherin expression was increased (all P<0.05). There were no relationship of the expressions of these proteins with patients’ age, gender, tumor location and degree of tumor differentiation (P>0.05). The increased expression of Wnt2b in bile duct cancer cells was related to increase in EMT marker of the positive expression of vimentin and the loss of E-cadherin expression (both P<0.05).@*Conclusions@#Wnt2b protein overexpression in cholangiocarcinoma correlated with epithelial-mesenchymal transition related markers. The Wnt2b protein was correlated with cholangiocarcinoma occurrence, development, invasion and metastasis. Wnt2b has the potential to develop into a new therapeutic target for carcinoma of bile duct.
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Objective To study the expression of Wnt2b protein and the epithelial-mesenchymal transition related markers in tissues of carcinoma of bile duct and normal bile duct to determine the clinical significance. The relationships between the expression levels and clinicopathological characteristics were analyzed, and the correlation between Wnt2b and epithelial interstitial transformation (EMT), tumor inva-sion and metastasis were studied. Methods A total of 60 patients with cholangiocarcinoma and 30 patients with normal bile duct tissues admitted to the Affiliated Hospital of Qingdao University from December 2008 to December 2013 were studied. The expressions of Wnt2b, E-cadherin and Vimentin protein were detected by SP immunohistochemical staining. The patients were classified according to the expressions of these proteins. Analyses were conducted on the relationships of these proteins with clinical characteristics of the patients with cholangiocarcinoma. Results The positive expression rate of Wnt2b protein in carcinoma of bile duct tissues was 90. 0％, which was significantly higher than that in normal bile duct tissues (χ2 =38. 1, P<0. 05). The positive expression rate of Wnt2b in T3 + T4 was significantly higher than that in T1 + T2(P<0. 05). There were no correlations of the expression with patients' age, gender, tumor location and degree of tumor differentiation (P>0. 05). Patients with lymph node metastasis had a significantly higher positive expression of Vimentin than patients with no lymph node metastasis, and the loss of E-cadherin expression was increased (all P<0. 05). There were no relationship of the expressions of these proteins with patients'age, gender, tumor location and degree of tumor differentiation (P>0. 05). The increased expression of Wnt2b in bile duct cancer cells was related to increase in EMT marker of the positive expression of vimentin and the loss of E-cadherin expression (both P <0. 05). Conclusions Wnt2b protein overexpression in cholangiocarcinoma correlated with epithelial-mesenchymal transition related markers. The Wnt2b protein was correlated with cholangiocarcinoma occurrence, development, invasion and metastasis. Wnt2b has the poten-tial to develop into a new therapeutic target for carcinoma of bile duct.
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Objective To evaluate the effect of melatonin pretreament on cell apoptosis and autophagy during lung ischemia-reperfusion (I/R) in rats.Methods Thirty-six adult male Sprague-Dawley rats,weighing 230-280 g,were divided into 3 groups (n=12 each) using a random number table method:sham operation group (group Sham),pulmonary I/R group (LIR group) and melatonin pretreatment group (MLT group).Lung I/R injury model was established by clamping the left hilum of lung for 60 min followed by 2 h reperfusion in LIR and MLT groups.Melatonin 1 mg/100 g was intraperitoneally injected at 15 min before clamping the left hilum of lung in group MLT.The rats were sacrificed at the end of reperfusion,the left lung was lavaged and the broncho-alveolar lavage fluid (BALF) was colleted for determination of protein concentrations.Lungs were removed for microscopic examination of the pathological changes (with a light microscope) which were scored and for determination of wet to dry weight ratio (W/D ratio),cell apoptosis (by TUNEL) and expression of Bcl-2,Bax,microtubule-related protein 1 light chain 3B Ⅰ (LC3B Ⅰ),LC3B Ⅱ,Beclin-1 and phosphorylated mammal rapamycin target protein receptor (p-mTOR) (using Western blot).The apoptosis index,Bcl-2/Bax ratio and LC3B Ⅱ/LC3B Ⅰ ratio were calculated.Results Compared with Sham group,the protein concentration in BALF,W/D ratio of lung tissues,apoptosis index,LC3B Ⅱ/LC3B Ⅰ ratio and Beclin-1 expression were significantly increased,and Bcl-2/Bax ratio and p-mTOR expression were decreased in LIR and MLT groups (P＜0.05).Compared with LIR group,the protein concentration in BALF,W/D ratio of lung tissues,apoptosis index,LC3B Ⅱ/LC3B Ⅰ ratio and Beclin-1 expression were significantly decreased,and Bcl-2/Bax ratio and p-mTOR expression were increased in MLT group (P＜0.05).Conclusion The mechanism by which melatonin pretreatment mitigates lung I/R injury may be related to inhibiting cell apoptosis and autophagy in rats.
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Objective@#To clarify the clinicopathologic features of hepatocellular carcinoma (HCC) patients survived more than 10 years after radical hepatectomy.@*Methods@#Two hundreds and fifty-two patients who underwent curative resection for HCC between January 1999 and March 2006 at Department of Hepatopancreatobiliary Surgery, Affiliated Hospital of Qingdao University were included.There were 217 male cases and 35 female cases aging from 17 to 82 years with median age of (53.8±10.5)years. Followed by March 31 2016, clinicopathologic factors in 10-year survivors and patients who died within 10 years were compared by χ2 test, Kaplan-Meier survival analysis and Cox proportional hazards model and the prognostic factors affecting survival were identified.@*Results@#All patients were followed-up for 4.0 to 205.7 months with median time of 53.4 months. The 10-year overall survival rate was 26%, there were 62 cases(26.2%) who survived for more than 10 years after initial hepatectomy. In survival >10-year group, the paitents with ALT<40 U/L, gamma-glutamyl transpeptidase<64 U/L, albumin≥35 g/L, without liver cirrhosis and portal hypertension, Child-Pugh grade A, no blood transfusion, AFP≤20 μg/L, tumor size ≤5.0 cm, single tumor, high differentiation, TNM stage Ⅰ and TACE negative after resection were more than the patients in survival <10-year group (P<0.05). In multivariate analysis, Child-Pugh grade A, the tumor size ≤5.0 cm and TACE negative after resection were favorable independent factors associated with 10-year survival (P<0.05).@*Conclusion@#Based on the results of the study, Child-Pugh grade A, tumor size ≤5.0 cm and TACE negative after resection at initial hepatectomy might be biologically favorable conditions for patients surviving more than 10 years.
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OBJECTIVE:To establish the method for the determination of plasma concentration of magnesium isoglycyrrhiz-inate in portal vein and peripheral venous blood of patients underwent liver resection,to further validate and evaluate pharmacoki-netic characteristics,rational and safe use of drugs in the clinic. METHODS:31 patients underwent liver resection in our hospital during Oct. 2014-Mar. 2015 were given magnesium isoglycyrrhizinate intravenously at the beginning of surgery. Portal vein and pe-ripheral venous blood of patients were drawn at 1 hour after drug use,and HPLC-UV detection method was used to determine the plasma concentration of drug. RESULTS:The retention time of isoglycyrrhizinate magnesium was 4.5 min,which showed a good peak shape,and was not interfered with the determination by plasma endogenous peak. The plasma concentration ranged from 0.55 to 55.00 mg/L. The minimum quantitative concentration was 0.55 mg/L. The extraction recoveries were 84.7%-87.1%,and method recoveries were 101.2%-105.4%,and RSDs of intra-day and inter-day were less than 6%. Plasma concentration of magnesium iso-glycyrrhizinate in portal vein blood was significantly higher than in peripheral vein blood of patients underwent liver resection (close to 2 times);and plasma concentration was not affected by primary liver diseases and underlying diseases such as cirrhosis. CONCLUSIONS:The method is simple and has high recovery rate of extraction,high accuracy and high sensitivity. It can meet the needs of pharmacokinetic study. After the application of magnesium isoglycyrrhizinate during liver resection,there is higher blood concentration of magnesium isoglycyrrhizinate in portal vein,which is beneficial to protect liver cells and improve liver func-tion. It is suitable during perioperative period of liver.
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Objective To compare the clinical result of Da Vinci robot-assisted distal pancreatectomy(RDP) and laparoscopic distal pancreatectomy(LDP), and to evaluate the clinical application experience of Da Vinci robotassisted spleen-preserving distal pancreatectomy.Methods From March 2013 to June 2015, totally 12 patients undergone RDP and 22 patients undergone LDP in our department were analyzed retrospectively.Results Intraoperative blood loss, hospitalization duration and postoperative fast time in RDP group was less than that in LDP group, the spleen-preserving rate and hospitalization expenses were higher in RDP group(P ＜ 0.05).There was no statistically significant difference in the rate of surgery duration and incidence of postoperative complication between two groups (P ＞ 0.05).The following-up period was 1-28 months with a mean of (9.94 ± 8.99) months, 1 case of peritoneal infection occurred in RDP group, 1 case of survival with tumor recurrence and 3 case suffered peritoneal infection in LDP group.Others were no metastasis, recurrence or death.Conclusions RDP is safe and feasible, the short-term prognosis is better than that of LDP.It has advantages of cleat 3 D visual field, stability in control,less invasive,and quick recovery.It is worth further clinical use.
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Objective To investigate the effect of palliative surgical treatment and minimally invasive biliary drainage of hilar cholangiocarcinoma.Methods Retrospectively collected 244 hilar cholangiocarcinoma patients clinical data in Affiliated Hospital of Qiingdao University between Jan.1,2008 to Dec.31,2011.Survival accoding to different treatment methods was compared using Kaplan-Meier method.The continuous measurement data were analyzed using the one-way ANOVA and the U test.The categorical variable were analyzed using the chi-square test or Fisher exact test.Result Among 244 patients, the R1/R2 resection group in 93 patients, the endoscopic retrograde biviar drainage group in 69 patients, the percutaneous transhepatic cholangial drainage group in 82 patiens.Three groups of median survival time and l-, 2-, 3-year survival rate and median survival time were (13.5 months, 8.9 months, 8.6 months), (63.0％, 24.7％, 7.4％), (33.3％, 3.3％, 0), (32.4％,4.2％, 0), respectively.There was significant difference in the survival time between R1/R2-resections and endoscopic retrograde biviary drainage treatments (P ＜ 0.001).R1/R2-resections and percutaneous transhepatic cholangial drainage treatments were statistical differences in survival time (P ＜ 0.001).Endoscopic retrograde biviary drainage and percutaneous transhepatic cholangial drainage treatments were no statistical differences in survival time (P =0.971).Conclusions Palliative surgical treatment for hilar cholangiocarcinoma patients had a more significant effect.Endoscopic retrograde biviary drainage and percutaneous transhepatic cholangial drainage of minimally invasive jaundice reducing method for prognosis in patients with hilar cholangiocarcinoma was same.
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ObjectiveTo study the effect of α7 ( α7 AChR) agonist nicotine on regulating sensitivity of regular chemotherapeutic agent in cholangiocarcinoma cells,and explore the possible target.MethodsThe effect of nicotine and α-BTX pretreatment on the survival ability of cholangiocarcinoma cells was investigated when applied with 5-FU by using MTT and Flat cloning formation experiment.ResultsApplied with 5-FU,in various con centrations nicotine stimulating group( 10-3 g/L,10-4 g/L,10-5 g/L ),the survive rate of QBC939 was 128％,124％,118％,while that in α-BTX stimulating group and combined stimulation group was 92％,94％,93％,92％,respectively.The cloning formation ability of nicotine- stimulating group (6.2 ± 0.40) was significantly higher than α- BTX stimulating group (3.2 ± 0.20 ),combined stimulation group ( 3.2 ± 0.20 ) and control group ( 3.4 ±0.33).ConclusionNicotine can prevent chemotherapy-induced apoptosis,and improve cholangiocarcinoma cell survival via α7 nicotine acetylcholine receptor in vitro.
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Objective To explore the experiment condition and method for the application of in vitro in vasive Transwell chamber and to observe muscarinicreceptor stimulant and muscarinicreceptor antagonist's influence to cholangiocarcinoma's invasiveness.Methods Two hundred microliter cell suspension of various concentrations(0.5×105/mL,1.0×105/mL,1.5×105/mL and 2.0×105/mL)was added into the upper chamber of the Transwell chamber,and the cells were allowed to penetrate the matrigel for 12,18,24and 48 hours respectively.The numbers was gotten as the invasive cells on the under surface of the membrane.After optimal cell concentration and time were gotten,pilocarpine of various concentrations(0 mmol/L,0.1 mmol/L,0.3 mmol/L and 0.5 mmoL/L)was added into the upper chamber of the Transwell chamber,then the cells on the matrigel were stained and counted.So did the cells when atropine of various concentrations(0.01 mmol/L,0.01 mmol/L,0.05 mmoVL and 0.1 mmol/L)were added into the upper chamher of the Transwell chamber in according to pilocarpine of various concentrations(0 mmol/L,0.3 mmol/L,0.3 mmol/L and 0.3mmol/L).Results With the increase of the time and cell concentrations，the cells couts that penetrated the matrigel increased，while the increase tended to he stable when the culture time exceeded 36 hous and the cell concentration Was over 1.0×105/mL.By adding pilocarpine,there were significant differences between the control and experimental groups(P＜0.05)，but there were no significant differences in experimental groups with various concentrations.There were no significant differences in blank group and experimental groups with atropine added(P＞0.05).When added pilocarpine and atropine,there were significant differences between blank and experimental groups(P＜0.05)，but there were no significant differences in experimental groups with various concentrations.Conclusions Thirty-six hours as invasive time,and one cell concentration 1.0 × 105/mL were optimal to test invasion abilities of cholangiocarcingma cells to different medicines or reagents.There is the possibility that museariniereceptor exists in cholangiocarcinoma cells，and may play an important role in cholangiocarcinoma's invasiveness and metastasis.
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Objective To investigate expression of slug and E-cadherin in pancreatic cancer tissues and determine the inhibitory effects of anti-Slug, an anti-sense plasmid, on the invasion of pancreatic cancer cell lines in vitro. Methods Slug and E-cadherin protein and mRNA was analyzed by IHP and RT-PCR in 36 cases of pancreatic cancer. Then anti-Slug plasmid was transfected into herin and Slug expression. The inhibitory effects of anti-sense Slug were also detected by Transwell motility assay and Matrigel invasion assay. Results The expression of Slug and mRNA in metastatic pancreatic cancer tissue was higher than that in non-metastatic tissue. E-cadherin and mRNA was lower in metastasis tissues(P＜0.05). The inverse relationships were further observed by transient transfection of anti-Slug into SW1990H4 cells. The downregulated expression of Slug and re-expression of E-cadherin were found. The Slug mRNA levels were 0.985±0.016,0.973±0.014, 0. 554±0. 011 after 0, 48 h of transfection of anti-sense Slug, and that of E-cadherin were 0.120±0.001, 0.360±0.002, 0. 727±0. 006, respectively. The diference was significant between different time points (P＜0.05). The Slug mRNA levels were 0. 206±0.017, 0.968±0.015, and that of E-cadherin were 0. 18±0.002,0.727±0.006 after stable transfection of anti-sense Slug, and control plasmid, respectively. The diference was significant (P＜0.05). The motility activity(393±28, 352±24, 96 ±13 )and the invasion activity (223 ± 69, 202 ± 64, 65 ±19) of1 antisense Slug transfectant cells were significantly decreased as compared with those of control cells (P＜0.05). Conclusions Higher expression of slug and lower expression of E-cadherin is related to the invasion and metastasis in pancreatic cancer. A reverse corelation of E-cadherin and Slug expression exists in pancreatic cancer. Slug is possibly a potential target for cancer gene therapy blocking invasion and metastasis in human pancreatic cancer.
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Objective Through studying the apoptosis induced by stichopus japonicus acid mucopoly saccharide in the hepatocellular carcinoma cell line HepG2 in vitro, analysing the expression of Bcl-2 and nm-23in HepG2, to provide the theory foundation and its feasibility on whether it can be used for the chemotherapy of hepetocellular carcinoma. Methods The cells of HepG2 were cultured in vitro and treated with SJAMP at different doses(0.25,0. 5,1.0,2.0,4.0 g/L). MTT was used to observe the inhibitory effects of SJAMP on cell growth, Western blotting was used to detect apoptosis, and the apoptosis related change of expression of protein Bcl-2 and nm23-H1. Results (1) MTT identified that SJAMP produced an obvious time-and-dose-dependent inhibitory effect on the HlepG2 cells. (2) Western blot showed that SJAMP could induce the apoptosis of HepG2 cells through changing the expression of the protein of Bcl-2 and nn23-H1 (P<0.05). Conclusion (1)SJAMP produced obvious inhibitory effects on HepG2 cells and induce HepG2 apoptosis. (2)SJAMP can enduce the anti-tumor function in the method of changing the expression of protein Bcl-2 and nm23-H1.
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Cholangiocarcinoma is a type of malignant tumor with high destruction.Due to its low diagnostic rate and high fatality rate,the operation is the unique therapeutic methods for the radical cure.However,the diagnosis and treatment for the disease were always in the phase of progression,so currently,the radical therapeutic rate is quite low,while the recurrence rate of the operation is extremely high.If the correlated mechanism of perineural invasion of cholangiocarcinoma could be understood,then interrupted its perineural invasion in the early period,that could greatly enhance the prognosis of cholangiocarcinoma patients.This article systematically reviews the progress of cholangiocarcinoma neural invasion related molecules and possible mechanism.
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Objective To explore the influence of PUMA on radiosensitivity of pancreatic cancer AsPC-1 cells after Slug gene inhibition by transfected short interferencing RNA(siRNA). Methods The AsPC-1 cells were infected with MOI 10,50,100 for 72 h, respectively. The expression of Slug and PUMA was analyzed by Western blotting and immunohistochemistry methods. The transfected and control cells were exposed to 4 Gy γ-rays. The cells inhibition rate was examined by MTT, Hoechst 33342 and IP double staining. DNA ladder and Giemsa staning was used to observe apoptosis. Results The relative value of Slug expression was 0.831 ±0.14,0. 546 ±0.12 and 0.178 ±0.08 after AsPC-1 was infected with Slug-siRNA ( MOI 10,50,100) for 72 h, significantly lower than that of control group ( F = 4. 992,P ＜ 0.05 ).The relative value of PUMA was 0. 325 ±0. 07,0. 593 ±0. 11 and 0. 978 ±0. 12, after AsPC-1 was infected with Slug-siRNA ( MOI 10,50,100) for 72 h, significantly higher than that of control group ( F = 4. 324,P ＜ 0. 05 ). The cell proliferation rate was ( 78.76 ± 9. 36 ) % in transfection combined with radiosensitivity group, significantly higher than that of transfection group [ ( 43.68 ± 6.71 ) % ] and radiosensitivity group alone [( 19.25 ± 3.72)% ] (F = 5.056, P ＜ 0.05). The apoptosis of transfection combined with radiosensitivity group was significantly higher than that of others. Conclusions Slug gene targeting siRNA could inhibit the expression of Slug, and consequently increase the activation of PUMA expression, and so enhance the radiosensitivity to γ-rays.
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Objective To investigate the expression changes of nuclear factor kappa B (NF-?B) and matrix metalloproteinase-9 (MMP-9) in the cultured hepatocellular carcinoma cells 9204 (HCC9204) transfected with inhibitory kappa B alpha(I?B-?)vector. Methods After pcDNA3-I?B-? vector and pcDNA3 were transfected into HCC9204 by lipofectamine method, Western-blot and RT-PCR analysis were used to detect the expressions of NF-?B and MMP-9. Migration and invasion of tumor cells were assayed by fundus membrane invaded by them. Results When pcDNA3-I?B-? was transfected into HCC9204, the expression of NF-?B was decreased at the protein level, and the expression of MMP-9 mRNA and the invision and metastasis ability of transfected cells were obviously decreased. Conclusion When the activity of NF-?B is inhibited, the ability of invasion and metastasis in HCC9204 cells decrease, which could be related to the decreased the expression of MMP-9.
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Objective To evaluate the efficacy of fibercholedochoscopy for the removal of residual stones after a surgical choledochostomy. Methods Two hundred and twenty cases of cholelithiasis underwent fibercholedochoscopy through a surgically formed T tube fistulae for residual stones from Sept. 1993 to Feb. 2002. Results A total of 572 times of fibercholedochoscopy was performed with residual stones totally evacuated in 201 cases (91.4%). Complications developed in 84 cases with no mortality. Conclusion Postoperative fibercholedochoscopy through a T tube fistulae is less traumatic and effective remedy for postoperatively retained common bile duct stones.
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ObjectiveTo evaluate the efficacy and feasibility of extraperitoneal approach (EPA) for the resection of primary retroperitoneal tumors (PRT). MethodsForty six cases undergoing resection of PRT were analyzed retrospectively, of which, 26 cases were through transabdominal approach (TAA group) and 20 through EPA. ResultsThe postoperative complications in EPA group was lower than in TAA group (2/20 vs. 11/26, P
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Objective To compare early enteral with parenteral nutrional support in patients after hepatectomy. Methods In this study, 59 patients were randomized into 2 groups to respectively receive enteral or parenteral nutritional support beginning the first day post-op for a week. The general nutrition condition, liver function, gut function, dosage of albumin, mortality, complication rate and expense were recorded. Results Patients were given same quantity of heat and nitrogen. At the end of the study, serum albumin, body weight and upper arm circumference had not reached the preoperative level in patients receiving enteral mutrition while all except for serum prealbumin had not reached the level in parenterally nutritional patients. Furthermore, the time of gut begins functional (29?12) h in enterally nutritional patients was shorter than in parenterally nutritional patients (38?14) h. Enteral nutrition was more economic than parenteral nutrition (P