Objective To compare the double-guide wire technique (DGT) with the standard cannulation technique (SCT) in patients with difficult access due to biliary complications after liver transplantation.Methods Difficult CBD cannulation is characterized by unsuccessful cannulation in 10 minutes.A total of 91 patients with biliary complications after liver transplantation were assigned to the DGT group (44patients,including 6 difficult cannulation,41 males and 3 females,30 to 61 years) and the SCT group (47patients,41 males and 6 females,33 to 56 years).An extra 20-minute cannulation was performed on the two groups.Success rate,procedure time and complications were compared.Results CBD cannulation was successful in 36 (81.8％) patients of DGT group and 33 (70.2％ ) patients of SCT group,which was not different ( P ＞ 0.05 ).The time of successful CBD cannulation in the DGT group ( 11.7 ± 3.2 minutes) was shorter than that in the SGT group ( 16.8 ±2.8 minutes,P ＜0.05).The incidence of post-ERCP hyperamylnsemia had no difference in the two groups ( P ＞ 0.05 ).There were no serious complications like infection,hemorrhage or perforation in either group.Mild pancreatitis occurred in 2 cases in the SCT group,but none in DGT.Conclusion DGT is an effective and safe technique in patients with biliary complications after liver transplantation,with no more complications than the SCT group.It is recommended in difficult cannulation of common bile duct (CBD).

ObjectiveTo prospectively study the safety and feasibility of sedation with propofol plus fentanyl for cirrhotic patients undergoing upper gastrointestinal endoscopy (UGIE).MethodsA total of 50cirrhotic patients and 50 control subjects without liver diseases referred to UGIE were assigned to the cirrhotic sedation group and the non-cirrhotic sedation group,respectively.Patients of both groups received sedation with propofol plus fentanyl.Meanwhile,30 cirrhotic patients underwent conventional UGIE.Vital signs of all subjects were recorded before sedation and procedure,five minutes,ten minutes and one hour after the procedure.Number connection test A (NCT-A) and line tracing test (LTT) were completed for all patients before sedation or procedures and 4 hours after endoscopy procedures.Occurrence of sedation-related complications was measured.ResultsIn the cirrhotic sedation group and the non-cirrhotic sedation group,blood pressure,heart rate,respiratory rate and saturation of pulse oximetry decreased of different degrees after secation (P ＞ 0.05 or P＜ 0.05),but returned to normal one hour after endoscopy procedures ( P ＞ 0.05).The total complication rates differed significantly between the cirrhotic sedation group and the non-cirrhotic sedation group [ 36％ (18/50) v.s.14％ (7/50),P ＜0.05 ].However,the rate of such complications as hypotension,bradycardia and hypoxemia in both groups was of no statistical difference (P ＞0.05 ).No cirrhotic patient developed overt hepatic encephalopathy after procedures.In addition,the NCT-A and LTT times before and after sedation in the cirrhotic sedation group and the cirrhotic non-sedation group were longer than those before and after procedure in the non-cirrhotic sedation group ( before sedation or procedure:(55.1 ±22.1)s,(58.6±23.1)s v.s.(36.9±7.0)s,(98.6±33.1)s,(89.5±15.6)s v.s.(81.4±13.6)s,P＜0.05; four hours after procedure:(54.4 ±21.6)s,(58.3 ±22.4)s v.s.(36.3 ±6.3)s,(88.4 ±30.6)s,(80.2 ±15.9)s v.s.(71.8 ± 12.0)s,P＜0.05,while there was no difference between cirrhotic sedation group and cirrhotic non-sedation group ( P ＞ 0.05 ).Within-group comparison showed NCT-A did not change ( P ＞ 0.05 ),whereas,LTT was obviously shorter than pre-sedation or pre-procedure ( P ＜ 0.05) due to learning effect.The differences in the NCT-A and LTT times before and after sedation or procedure were not significant among the three groups (P ＞ 0.05 ).ConclusionSedation with propofol plus fentanyl is relatively safe in cirrhotic patients during UGIE,which will not precipitate hepatic encephalopathy or cause irreversible complications.

Objective To study the effect of matrine on the expression of a proliferation-inducing ligand (APRIL) in colorectal cancer cell line (SW480 cell). Methods MTT assay was used to evaluate the inhibitory effect of matrine on SW480 cells. The protein and mRNA levels of APRIL in SW480 cells were determined by immunohistochemistry and real-time fluorescence quantitative PCR (RFQ-PCR). SW480 cells were treated with 0.5,1.0,2.0 mg/ml of matrine for 24 h, 48 h and 72 h. FU and blank were served as drug control and blank control groups, respectively. Results Matrine had obviously inhibitory effect on proliferation of SW480 cells in a time- and dose-dependant manner. The expression of APRIL was strong in SW480 cells. When treated with 50,100,200 ug/ml of FU, the APRIL mRNA levels in SW480 cells raised gradually and reached the highest levels at 72 h after treatment, which were significantly higher than those in blank control group (all P value<0.001). When treated with 0. 5,1.0, 2.0 mg/ml of matrine, the APRIL mRNA levels in SW480 cells increased at 24 h after treatment, which were significantly higher than those in blank control group (all P value<0. 001), and then decreased gradually and almost equal to level of blank control group at 72 h. Conclusion In treatment with FU, the survival cells.may have stronger ability of proliferation due to higher expression of APRIL in SW480 cells. Anti-APRIL therapy might be an important assistant treatment to counter the impact of APRIL. Matrine will not cause persistent increase of APRIL mRNA levels in SW480 cells, so it might be a helpful drug in anti-tumor theraphy.

Objective To investigate the role of TLR4 in the pathogenesis of chronic hepatitis B(CHB) by study the expression of TLR4 in liver tissues in patients with CHB, and the relationship among TLR4 and serum HBV DNA level, clinical severity degrees and histo-logical grades and stages. Methods Expression of TLR4 in liver tissues was semi-quantitatively determined by immunohistochemistry and e-valuated by a scoring system in 75 patients with CHB and 10 health controls. Results The positive staining of TLR4 mainly located in the cytoplasm and some on cell membrane of bepatocytes. Expression of TLR4 in the liver tissues of patients with CHB was stronger than that of health controls. The scores of TLR4 expression in patients with mild, moderate and severe CHB were 1.0±0.5,2.3±0.5 and 2.9±0.4. The scores increased gradually and significantly along with the increase of clinical severity degrees( F = 104.8, P<0.01). The scores of TLR4 expression in the liver tissues of patients with CHB were positively correlated with the clinical severity degrees (r=0.838, P<0.01) and histological grades (r=0.579, P<0.05), but not correlated with Lg (serum HBV DNA) or histological stages. Conclusion TLR4 was up-regulated in the hepatocytes of patients with CHB. There may be a role of TLR4 in the pathogenesis of CHB.

Objective To observe the effects of 5-fluorouraeil(5-FU)and eisplatin(DDP)on the expression of Toll-like receptor 2 (TLR2)and Toll-like receptor4(TLR4)in hepatocellular carcinoma cell lines HepG2 and HepG2.2.15.Methods Direct immanotlaorescenee flow cytometry was used to detect mean flubrescence intensity(MFI)of TLR2 and TLR4,and TLR2 and TLR4 positive cell percentage in HepG2 and HepG2.2.15 cells before and after treated with 5-FU.and DDP at various concentrations for 24h,48h and 72h.Results MFI of TLR2 and TLR4.and TLR2 and 11LR4 positive cell percentage in HepG2.2.15 cells were significantly higher than those in HepG2(P＜0.01).After HepG2 and HepG2.2.15 cells were treated with different concentration of 5-FU and DDP,MFI of TLR2 and TLR4,TLR2 and TLR4 positive cell percentage in HepG2 and HepG2.2.15 cells almost had no change.only MFI of TLR2 in HepG2.2.15 cells decreased after cells were treated with 5-FU at the concentrations of 100,200μg/ml and DDP at the concentrations of 20μg/ml for 72h(P＜0.05 for all).Conclusions 5-FU and DDP can not activate TLR2 and TLR4 signal pathway in hepatocellular carcinoma cell lines HepG2 and HepG2.2.15.To find the activated pathway in TLR2 and TLR4 signal pathway,some other methods should be used,and this will be helpful in antieancer therapy.

Objective To study the change of TLR4 on peripheral blood monoeytes (PBMCs) and its role in the pathogenesis of chronic severe hepatitis B. Methods The expression of TLR4 on 10000 CDI4 + PBMCs was determined by flow eytometer in 30 healthy control,31 patients with chronic hepatitis B and 30 patients with chronic severe hepatitis B. The level of serum tumor necrosis factor α(TNF- α) was determined by ELISA. Results The values of TLR4 on PBMCs and serum TNF-αof the groups of healthy control, patients with chronic hepatitis B and patients with chronic severe hepatitis B were 2.3±1.1,3.7±2.3, (6.9±4.1 ) mean fluorescence intensity (MFI) and (53.8±38.1 ), ( 164.3±89.9) and (359.8±140.0) ng/L. The TLR4 value in patients with chronic severe hepatitis B was signifi- cant higher than those in healthy control and the patients with chronic hepatitis B ( P <0.05). However, there was no significant difference between the patients with chronic hepatitis B and healthy control ( P > O. 05 ). TNF-α increased gradually and significantly from the healthy control to the patients with chronic hepatitis B and patients with chronic severe hepatitis B. There was a significant positive correlation be- tween the value of TLR4 and the value of serum TNF-αin the patients with chronic severe hepatitis B( r=0.666, P <0.01). Conclusion There may be a role of TLR4 in the pathogenesis of chronic severe hepatitis B.

Objective:To observe the clinical effect of Bairui grain combined with ambroxol atomization inhalation treating acute attack of chronic bronchitis (wind heat attact the lung symptom). Methods:A total of 150 patients with acute attack of chronic bronchitis who were admitted in our hospital from July 2015 to July 2018 were randomly divided into control and intervention groups (75 each group) by the random number table method. The control group received ambroxol atomization inhalatio based on the regular western medicine treatment; the intervention group took Bairui grain orally based on control group. Both of the two groups were treated for 2 weeks. Before and after the treatment, to score the clinical symptoms and wind-heat attact the lung symptom. Use pulmonary function meter to detect the ratio of FEV1 to estimated value (FEV1%) and FEV1/FVC; use ELISA to detect serum and sputum levels of TNF-α and IL-6. Record the extinction time of the symptoms (cough, wheezing, sputum, lung rumble) and evaluate the clinical efficacy. Results:The total efficacy rate of intervention group was 98.6% (72/73), which was significantly higher than control group 88.9% (64/73) ( χ2=4.354, P=0.037). The symptoms (cough, wheezing, sputum, lung rumble) extinction time of intervention group were significantly less than those of the control group ( t values 5.331, 5.590, 5.841, 6.305, respectively, all Ps<0.01). After the treatment, scores of clinical symptoms (cough, wheeze, cough, cough) and symptoms of wind-heat attacking lung (cough, wheezing, fever, dry mouth, stuffy nose, runny nose) of intervention group were significantly lower than those of the control group ( t values 4.990, 4.431, 5.221, 5.004, 5.652, 5.190, 5.311, 5.793, 5.643, respectively, all Ps<0.01). After treatment, the FEV1% (52.51% ± 5.63% vs. 47.30% ± 5.21%, t=8.931) and FEV1/FVC (61.57 ± 6.44 vs. 56.87 ±5.82, t=8.251) were significantly higher than the control group ( P<0.01). The serum level of TNF-α, IL-6 ( t values 5.331, 4.908) and the level of TNF-α, IL-6 ( t values 6.001, 4.803) in sputum were significantly lower than those of the control group ( P<0.01). Conclusions:The Bairui grain combined with ambroxol atomization inhalation can decrease the inflammotory cytokine levels of the acute attack patients with chronic bronchitis (syndrome of wind-heat attacking lung symptome), improve clinical symptoms and enhance efficacy.

ObjectiveTo investigate the application value of endoscopic retrograde cholangiopancreatography (ERCP) in biliary stricture following liver transplantation (LT).MethodsClinical data of 47 patients with biliary stricture following LT treated with ERCP in the Third Affiliated Hospital of Sun Yat-sen University between January 2008 and January 2014 were retrospectively studied. The informed consents of all patients were obtained and the local ethical committee approval had been received. Among the 47 patients, 41 were males and 6 were females with the age ranging from 24 to 62 years old and the median of 51 years old. Anastomotic stricture was observed in 31 cases and non-anastomotic stricture in 16 cases. ERCP was performed on patients to localize biliary stricture and then papillotomy was performed under the support of guide wire. The stricture was dilated progressively by dilating catheter or columnar balloon through the guide wire. Endoscopic nasobiliary drainage or plastic stent placement was chosen according to the ERCP results. The evaluation of curative effect was graded by cure, improvement, inefifcacy and restenosis.ResultsAll the 47 patients underwent ERCP successfully with totally 112 times. Nasobiliary drainage was performed 79 person-times, stent placement 33 person-times. Among the 31 patients with anastomtic stricture, 28 were cured and 3 were improved. Among the 16 patients with non-anastomtic stricture, 5 were cured, 3 were improved, and 8 were ineffective. The total cured rate of all the patients was 70% (33/47) and the effective rate was 83% (39/47). The incidence of complications was 9% (4/47), including 2 cases of mild pancreatitis, 1 case of hyperamylasemia and 1 case of biliary tract infection.ConclusionERCP is safe, effective and integrated in diagnosis and treatment for biliary stricture following LT, which is the ifrst choice for non-surgery treatment.

ObjectiveTo investigate the changes and role of β-arrestin 1 in the course of non-alcoholic fatty liver disease (NAFLD) progressing to hepatocellular carcinoma (HCC). MethodsEighty healthy male C57BL/6 mice were randomized into the vegetarian diet group and the high fat diet group according to the random number table with 40 mice in each group. Mice in the vegetarian diet group were fed with vegetarian diet (13% calories in fat) and mice in the high fat diet group were fed with high fat diet (58% calories in fat). Eight mice in each group were decapitated at the end of 9 and 24 weeks. The rest mice in each group were decapitated at the end of 48 weeks. The incidence of HCC of two groups was observed. The expression of proteinβ-arrestin 1 in the liver tissues of mice was detected by Western blot and the mRNA level was examined using TaqMan real time fluorescence quantitative RT-PCR. The incidence of HCC in two groups was compared using Fisher's exact test, and the protein β-arrestin 1 expression and mRNA level of two groups were compared usingt test. Spearman correlation analysis was used to evaluate the relationship between protein β-arrestin 1 expression, mRNA level and the feeding duration of high fat diet in high fat diet group.ResultsThe incidence of HCC in the high fat diet group was 18% (4/22), which was significantly higher than 0 (0/23) in the vegetarian diet group (P=0.034). The expression level of protein β-arrestin 1 in liver tissues of mice in the high fat diet group was 2.4±0.5 in the 9th week, which was significantly higher than 1.5±0.4 in the vegetarian diet group (t=2.779,P<0.05). The β-arrestin 1 mRNA level in liver tissues of mice in the high fat diet group in the 9th, 24th and 48th week were 4.1±0.8, 7.8±2.1 and 12.5±1.2 respectively, which were all significantly higher than 2.6±0.7, 3.6±0.6 and 6.9±1.2 in the vegetarian diet group (t=4.029, 5.522, 9.487;P<0.05) . The protein β-arrestin 1 and mRNA level in HCC tissues of mice in the high fat diet group in the 48th week were 4.6±0.5 and 22.0±3.2, which were signiifcantly higher than 1.6±0.4 and 12.5±1.2 in liver tissues at the same period (t=9.600, 7.837;P<0.05). The protein β-arrestin 1 and mRNA level in high fat diet group were positively correlated with the duration of high fat diet (r=0.949, 0.922;P<0.05). Conclusions It is likely to develop NALFD for the mice fed with high fat diet, and the incidence of HCC is signiifcantly increased. β-arrestin 1 may play a role of accelerating the course of NAFLD progressing to HCC.

ObjectiveTo explore the influence of portal vein thrombosis (PVT) on the clinical efifcacy of endoscopic esophageal variceal ligation (EVL) in cirrhotic patients with esophagogastric variceal hemorrhage (EVH).MethodsClinical data of 314 cirrhotic patients with EVH who underwent endoscopic EVL and were followed up for more than 6 months in the Third Afifliated Hospital of Sun Yat-sen University between January 2005 and December 2014 were retrospectively analyzed. The informed consents of all patients were obtained and the local ethical committee approval was received. The patients were divided into the PVT group and the control group according to whether they had PVT during treatment. Among the 72 patients in the PVT group, 61 were males and 11 were females with the mean age of (50±11) years old.Among the 242 patients in the control group, 206 were males and 36 were females with the mean age of (47±11) years old. The clinical efifcacy of two groups was compared, and the correlation between PVT and the elimination rate of esophageal varices (EV) as well as the recurrent bleeding rate of EV was analyzed. The number of EVL treatment period in two groups was compared usingt test and the rate was compared using Chi-square test. Univariate logistic regression analysis was conducted for the correlation between PVT and the elimination rate, recurrent bleeding rate of EV.ResultsThirty-six patients in the PVT group and 115 patients in the control group developed acute EVH. After EVL, the emergency hemostatic rate of both groups was 100%. The elimination rate of EV in the PVT group was 76%(55/72), signiifcantly lower than 90%(218/242) in the control group (χ2=9.166,P<0.05). The number of EVL treatment period in the PVT group was 3.4±1.6, significantly more than 2.8±1.1 in the control group (t=3.065,P<0.05). The recurrent bleeding rate of EV in the PVT group was 36%(26/72), signiifcantly higher than 21%(51/242) in the control group (χ2=6.779,P<0.05). PVT was a risk factor for both the elimination rate of EV (OR=0.356, 95%CI: 0.179-0.709,P<0.05) and the recurrent bleeding rate of EV (OR=2.383, 95%CI: 1.354-4.196,P<0.05). ConclusionPVT is a risk factor for both the elimination rate of EV and the recurrent bleeding rate of EV in cirrhotic patients with EVH treated by endoscopic EVL.