Objective:To explore the mechanism of biliary fibrosis after end to end anastomosis of bile duct.Methods:12 Neijiang miniature pigs (6-8 months old, weight 30-40 kg) were divided into experimental group ( n=6) and control group ( n=6). The model of end to end anastomosis after transection of the common bile duct was established in experimental group. Control group was a sham operation group, and only T tube was placed. After 9 months, Masson staining, fluorescence quantitative PCR and immunohistochemistry were used to analyze the expressing changes of pro-fibrotic factor transforming growth factor-β1 (TGF-β1), connective tissue growth factor (CTGF) and epithelial-mesenchymal transition (EMT) markers, including cytokeratin-19 (CK19), E-cadherin (E-Cad) and fibroblast specific protein-1 (S100A4), α-smooth muscle actin (α-SMA) and collagen components Collagen I (COL-1), collagen III (COL-3) and fibronectin (FN) in the anastomotic bile duct tissues. Results:Masson staining showed that the submucosal collagen fibers increased significantly in the experimental group. Compared with the control group, the mRNA expression of TGF-β1 [(3.482±0.313) vs. (1.000±0.102), t=18.43, P<0.001], CTGF [(2.160±0.287) vs. (1.000±0.103), t=9.32, P<0.001] were increased, the difference was statistically significant. Compared with control group, the mRNA and protein expression of CK19 and E-Cad were decreased in the experimental group, while the mRNA and protein expression of S100A4 and α-SMA were increased in the experimental group (all P<0.01). Conclusion:It was feasible in the short term to perform an end-to-end anastomosis after transection of the common bile duct, but there was an obvious fibrosis in the anastomotic bile duct tissue at later time.

Hepatocellular carcinoma (HCC) with bile duct tumor thrombus (BDTT) is rare and enhanced CT or MRI can be used for its diagnosis. Surgical procedure is the main treatment for HCC with BDTT. The authors introduce the experiences of recurrent patient with HCC and BDTT who was treated with targeted therapy plus immunotherapy, in order to provide reference for its clinical diagnosis and treatment.

Objective:To evaluate liver quadrate lobectomy combined with large-bore cholangio-jejunostomy for the treatment of benign biliary-enteric anastomotic stricture.Methods:The clinical data of 32 patients undergoing liver quadrate lobectomy combined with large-caliber cholangio-jejunostomy were retrospectively analyzed. The bile drainage effect and postoperative complications were analyzed. And the clinical effects of different size of biliary-enteric anastomosis were compared.Results:The most common short-term complications were cholangitis (9.4%) and bile leakage (9.4%), and the most common long-term complications were reflux cholangitis (15.6%). No anastomotic restenosis, stone formation or canceration were found. During the follow-up period, the total bilirubin ( t=19.455, P=0.000), direct bilirubin ( t=18.479, P=0.000), alkaline phosphatase ( t=3.229, P=0.002) and γ-glutamyltranspeptidase ( t=3.057, P=0.003) level were significantly improved. The effect of bile drainage in patients with 2-3 cm diameter of biliary-enteric anastomosis was similar to that in patients with >3 cm diameter ( t=0.284, P=0.778). The incidences of cholangitis (χ 2=0.121, P=0.728), bile leakage (χ 2=0.121, P=0.728) and reflux cholangitis (χ 2=0.205, P=0.652) were no statistical difference in both groups. Conclusion:Liver quadrate lobectomy combined with large-caliber cholangio-jejunostomy is effective in the treatment of benign biliary-enteric anastomotic stricture. More than 2cm in diameter of the biliary-enteric anastomosis is enough.

Objective To investigate the effect and its molecular mechanism of phosphoglycerate mutase 5 (PGAM5) mediated pyroptosis on liver ischemia-reperfusion injury (IRI). Methods C57 mouse models of liver IRI were established and randomly divided into the 6 h reperfusion (6 h group) and 12 h reperfusion (12 h group), and sham operation group (sham group) was established too, 10 rats in each group. The effect of IRI on the parameters in the liver tissues and serum samples was evaluated. The expression levels of PGAM5 and cysteinyl aspartate specific proteinase (Caspase)-1 in the liver tissues during IRI were quantitatively detected. The IRI models of liver cells were established (IRI group). The IRI models of liver cells were established after pretreatment with Caspase-1 inhibitor Z-YVAD-FMK (inhibitor group). The untreated AML12 cells were allocated into the control group. The effect of inhibiting Caspase-1 activity on pyroptosis was analyzed. AML12 cells were transfected with PGAM5 small interfering ribonucleic acid (siRNA) (siRNA group) and siRNA-negative control (siRNA-NC) (siRNA-NC group) by liposome 3000, and then IRI models of liver cells were established. The untreated AML12 cells were assigned into the control group. The effect of PGAM5 mediated pyroptosis on IRI of liver cells was assessed. Results In the 6 h and 12 h groups, partial liver cell edema, hepatic sinusoid narrowing, central vein congestion and occasional spot necrosis were observed in the mouse liver tissues, and these changes in the 12 h group were more aggravated than those in the 6 h group. The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the 6 h and 12 h groups were higher than those in the sham group, and the values in the 12 h group were higher than those in the 6 h group. The levels of tumor necrosis factor (TNF)-α and interleukin (IL)-1β were increased in the 6 h and 12 h groups, and the values in the 12 h group were lower than those in the 6 h group. The relative expression levels of IL-1β messenger ribonucleic acid (mRNA) in the mouse liver tissues in the 6 h and 12 h groups were up-regulated, and the value in the 12 h group was lower than that in the 6 h group. The cell apoptosis rates in the liver tissues were significantly increased in the 6 h and 12 h groups, and the value in the 12 h group was remarkably lower than that in the 6 h group (P < 0.01-0.05). Compared with the sham group, the relative expression levels of PGAM5 mRNA and protein in the mouse liver tissues in the 6 h and 12 h groups were significantly up-regulated, and the values in the 12 h group were significantly higher than those in the 6 h group (P < 0.01-0.05). The protein expression levels of PGAM5 and Caspase-1 in the liver tissues were up-regulated in the 6 h and 12 h groups. Compared with the control group, the relative expression levels of NOD-like receptor protein 3 (NLRP3), cleaved Caspase-1 and Gasdermin D (GSDMD) proteins were up-regulated and the fluorescence intensity of GSDMD was increased in the IRI group. Compared with the IRI group, the relative expression levels of NLRP3, cleaved Caspase-1 and GSDMD proteins were significantly down-regulated and the fluorescence intensity of GSDMD was considerably decreased in the inhibitor group (P < 0.01-0.05). Compared with the control group, the cell survival rate was significantly decreased, and the relative expression levels of PGAM5, NLRP3, cleaved Caspase-1 and GSDMD proteins were significantly up-regulated in the siRNA-NC group (P < 0.01-0.05). Compared with the siRNA-NC group, the cell survival rate was remarkably increased, whereas the relative expression levels of PGAM5, NLRP3, cleaved Caspase-1 and GSDMD proteins were significantly down-regulated in the siRNA group (P < 0.01-0.05). Conclusions PGAM5 may aggravate the liver IRI in mouse models probably by mediating pyroptosis via PGAM5/Caspase-1/GSDMD signaling pathway and aggravating liver cell injury.

Objective:To investigate the effect of expanded lymphadenectomy on the survival rate of the patients with hilar cholangiocarcinoma(HC).Methods:The clinical data of 129 patients undergoing radical resection of HC were retrospectively analyzed. According to the range of lymphadenectomy, they were divided into regional group(91 cases) and expanded group(38 cases). The clinical data, survival rate and postoperative complications in two groups were compared and analyzed.Results:The 3-year and 5-year survival rates of metastasis free(M0) patients in the regional group and expanded group were 44.1%, 24.8% and 47.0%, 33.6%, respectively; there were no statistically significant difference in survival rates between the two groups(χ 2=0.662, P=0.416). Compared with patients in the regional group, the average number of harvested lymph nodes in the expanded group was significantly increased and the difference was statistically significant( t=14.678, P=0.000), while the incidence of severe complications and mortality does not increase significantly. Conclusion:Expanded lymphadenectomy did not improve the survival rate of M0 HC patients, which while yielding more lymph nodes does not increase the incidence of postoperative complications and mortality in resectable HC patients.

Objective To analyze the clinical characteristics,diagnosis,treatment and prognostic factors of primary hepatic neuroendocrine tumor (PHNET).Methods To analyze the clinical data of 21 patients with PHNET who were admitted to the First Affiliated Hospital of Zhengzhou University from January 2012 to January 2019.There were 11 males and 10 females,with ages which ranged from 36 to 75 years.Log-rank test was used for single-factor analysis of the relationship between clinical and pathological indicators and prognosis.Cox proportional risk model was used for multi-factor analysis.Results Among the 21 PHNET patients,10 presented with central and upper abdominal pain,3 abdominal pain accompanied by nausea and vomiting,5 abdominal distension,and 3 were asymptomatic.Carbohydrate antigen (CA) 19-9 was elevated in 6 patients and CA125 in 7 patients.Abdominal enhanced CT showed solid liver space occupying lesions.Four patients had solitary and 17 had multiple lesions.The mean diameter was 58 mm.Fourteen patients had lymph node metastasis.Five patients underwent radical hepatectomy,2 transcatheter hepatic arterial chemoembolization + chemotherapy,12 chemotherapy,and 2 supportive treatment.Factors which were associated with prognosis of PHNET patients were surgery,tumor grading,cytokeratin positivity and lymph node metastasis.Multivariate Cox regression analysis showed that inoperability (HR =8.99,95％ CI:1.13-71.80) was an independent risk factor of prognosis of PHNET patients.The prognosis in patients who underwent surgical resection was better.Conclusion Patients with PHNET had no specific clinical manifestations.Surgical resection gave the best results in treatment.As surgical resection affected prognosis,it should be carried out if technically feasible.

Objective:To investigate the efficacy of two different percutaneous transhepatic biliary drainage (PTBD) procedures for malignant obstructive jaundice.Methods:The clinical data of 132 patients with malignant obstructive jaundice who underwent PTBD were retrospectively analyzed. According to whether the tip of the catheter was placed in the intestine, 132 patients were divided into the external drainage group and the internal-external drainage group. The effect on decrease in jaundice (total bilirubin, direct bilirubin levels), postoperative complications, and hospitalization costs of the two drainage methods were compared.Results:Of the 132 patients who were included in this study, there were 91 males and 41 females, aged 18 to 85 years. The external drainage group included 81 patients, and the internal and external drainage group included 51 patients. The total bilirubin [(190.2±41.8)μmol/L vs. (294.9±38.3) μmol/L] and direct bilirubin [(155.4±30.9)μmol/L vs. (242.1±39.6) μmol/L] levels in the external drainage group and the total bilirubin [(179.3±37.1)μmol/L vs. (288.1±35.4)μmol/L] and direct bilirubin [(147.7±32.5)μmol/L vs. (233.7±36.1)μmol/L] levels in the internal-external drainage group were significantly decreased after surgery (all P<0.05). The incidences of reoperation, re-intubation or bilateral catheterization [15.6%(8/51) vs. 3.7%(3/81)] and biliary tract infection [50.9%(26/51) vs. 27.1%(22/81)] in the internal-external drainage group was significantly higher than that in the external drainage group (all P<0.05). The mortality rate due to biliary tract infection in the internal-external drainage group was significantly higher than that of the external drainage group [7.8%(4/51) vs. 0, P<0.05]. Intestinal-derived bacteria such as Escherichia coli, Enterococcus faecium, Pseudomonas aeruginosa, and Klebsiella pneumoniae were the main bacteria in the biliary tract infections of the two groups. The hospitalization cost of patients in the internal-external drainage group was significantly higher than that in the external drainage group [(34 928.0±3 693.0) yuan vs. (29 360.0±3 219.0) yuan, P<0.05]. Conclusion:Both PTBD external drainage and internal and external drainage could alleviate the symptoms of jaundice quickly and effectively.

Objective:To investigate the effect of local antibiotics irrigation on chronic proliferative cholangitis (CPC).Methods:CPC model of rabbits was established. Rabbits were divided into CPC group (without local antibiotics irrigation, n=20) and experimental group (with local antibiotics irrigation, n=20). Only the gallbladder was removed, and 20 rabbits with free bile duct (sham operation) were used as normal control. The inflammatory conditions, proliferation of cholangiocytes, biliary fibrosis and biliary stones formation ability were analyzed. Results:Compared with CPC group, the relative expression of inflammation index lipopolysaccharide and interleukin-6 [(1.21±0.13) vs. (3.24±0.21), (1.52±0.22) vs. (3.10±0.23)], biliary cell proliferation index cyclooxygenase-2 and vascular endothelial growth factor [(2.15±0.12) vs. (4.07±0.22), (2.44±0.14) vs. (3.22±0.21)], fibrosis index transforming growth factor-β and Collagen-I [(2.44±0.28) vs. (4.36±0.44), (1.54±0.13) vs. (2.22±0.18)] and biliary stones formation index β-glucuronidase and Mucin 5AC [(1.74±0.20) vs. (3.42±0.31), (1.47±0.15) vs. (2.81±0.22)] were significantly decreased in experimental group (all P<0.05). Conclusion:Local antibiotics irrigation could inhibit CPC by inhibiting the chronic inflammation of the biliary tract and excessive proliferation of cholangiocytes and biliary fibrosis, and reducing the probability of biliary stone formation.

Objective:To study the impact of additional resection of an intraoperative proximal bile duct positive margin on the survival rate of patients with hilar cholangiocarcinoma (HCCA).Methods:The clinical data of 214 patients with HCCA treated at the First Affiliated Hospital of Zhengzhou University from January 2000 to January 2017 were analyzed. There were 126 males and 88 females, aged (62.3±17.1) years. These patients were divided into three groups according to the pathological status of bile duct margins and whether the proximal bile duct was further resected. Group A consisted of patients with negative margins without additional resection (161 cases). Group B consisted of patients with negative margins after additional resection (21 cases). Group C consisted of patients with R 1 resections (32 cases). The Kaplan-Meier method was used for survival analysis and log-rank test was used for inter-group comparison. Univariate and multivariate Cox regression analysis were used to analyze prognostic factors. Results:The 3-year and 5-year survival rates of patients in groups A, B, and C were 37.1%, 28.6%, 0 and 18.5%, 10.7%, 0, respectively. The cumulative survival rates of patients in group A and group B were significantly higher than that in group C (all P<0.05). Multivariate Cox regression analysis showed that R 1 proximal bile duct margin ( HR=3.728, 95% CI: 2.531-4.936), margin width >5 mm ( HR=0.534, 95% CI: 0.224-0.857), and T 3-4 staging ( HR=5.655, 95% CI: 3.174-8.203) were independent influencing factors for overall survival of patients with HCCA after attempted radical surgery. Conclusion:The survival rate of patients with HCCA with a positive proximal bile duct margin was significantly improved by further resecting the bile duct to obtain a negative margin. The R 1 proximal bile duct margin was an independent risk factor for prognosis in patients with HCCA after attempted radical resection.