Kupffer cells, the resident macrophages of the liver, play an important role in ischemia/reperfusion(L/R)injury in liver transplantation. 1schemic liver is reperfused through portal vein, and then Kupffer cells are activated. The liver is damaged by the released reactive oxygen species, inflammatory mediators and cytokines. On the other hand, upregulation of HO-1, induced by the activated kupffer cells, exerts a cytoprotective function for the IYR injury in liver transplantation models. Thus, Kupffer cells play the dual role on L/R injury in liver transplantation.

Hepatic ischemia-reperfusion injury,a relatively common phenomenon in clinical pathophysiology, is one of the currently difficult and hot issues in liver surgery and liver transplantation clinical studies. p38 mitogen activated protein kinases are activated in a variety of diseases and stress conditions. p38 signaling pathway activation is one of the important mechanisms leading to hepatic ischemia-reperfusion injury. This article reviews the roles and mechanisms of P38 signaling pathway recently years in the liver ischemia reperfusion injury.

Functional insulinoma accounts for 85% of insulinoma, and it is evenly distributed in the head, body and tail of the pancreas. The main clinical manifestation of patients with functional insulinoma is endocrine disorder, and 92% of them presented with neurological symptoms. Preoperative localization of functional insulinoma is difficult because of the small size of the tumor. A 31-year-old male patient with the chief complaint of paroxysmal dizziness and confusion was admitted to the First Affiliated Hospital of Kunming Medical College on May 3,2010. The patient was preliminarily diagnosed with functional insulinoma by detecting the levels of fasting blood glucose,serum insulin and fasting serum C-peptide, as well as the presence of Whipple's triad. Ultrasonography and enhanced computed tomography demonstrated that a well-defined tumor of 13.0 mm ×13.0 mm in size was located in the pancreatic uncinate process.On May 27, 2010, the patient received surgical resection of the tumor, and histological examination of the resected specimen confirmed insulinoma.

Objective To investigate theeffect of ischemic postconditioning (IPO) on ischemia/reperfusion injury via mitochondrial pathway.Methods Male Sprague-Dawley rats were divided into three groups by means of random number table.Rats in IRI group and IPO group received liver transplantation.The portal vein in IRI group was opened immediately after liver implantation to restore the blood supply.The graft rats in IPO group received IPO before the portal vein was completely opened: 60-s ischemia and 60-s reperfusion of the portal vein,repeated 6 times).The rats in the sham-operation group were only subject to dissociation of the liver ligament.Six h after portal vein reperfusion,liver function was tested.The expression of Bcl-2 mRNA and Bax mRNA was detected by using real time-PCR.The protein expression of Cyt-c was detected by using Western blotting.The apoptosis and necrosis of liver tissue and the mitochondrial transmembrane potential were detected by using flow cytomertry.The changes of mitochondrial structure were observed under an electron microscope.Results The liver functions in IRI group and IPO group were significantly worse (P＜0.05),the levels of Bax mRNA were significantly higher (P＜0.05),the levels of Bcl-2 mRNA were significantly lower (P＜0.05),the levels of Cyt-c protein were significantly higher (P＜0.05),the levels of apoptosis and necrosis were significantly higher (P＜0.05),and mitochondrial transmembrane potential (MTP) of liver cells was significantly lower (P＜0.05) than in sham operation group.The changes of all the parameters in IRI group were more significant than in IPO group (P ＜ 0.05).The morphological changes of the liver mitochondria were also significantly aggravated in IRI group as compared with IPO group.Conclusion IPO reduced IRI by inhibiting the mitochondrial signaling pathway in rats undergoing liver transplantation.

Objective To explore the effect of heme oxygenase-1 (HO-1) on expressions of hypoxia inducing factor 1 alpha (HIF-1α) and vascular endothelial growth factor (VEGF)and regeneration of hepatic vascular plexus after orthotopic liver transplantation ischemia-reperfusion injury in rats.Methods Theexperimental study was conducted.According to the random number table,240 SD rats were divided into the 3 groups,80 rats in each group.Empty virus group:rats were transfected with the empty virus.Induced group:rats were transfected with HO-1 overexpression adenovirus.Inhibited group:rats were transfected with HO-1 RNAi adenovirus.Rats were made pairs (1 ∶ 1) and established rat liver transplantation model according to two cuffs method.Rats with less weight and with heavier weight were respectively chosen as donor rats and recipient rats,and then recieved tail intravenous injection of adenovirus at 24 hours before operation.(1) Detection of transfection efficiency of adenovirus before operation:HO-1 expression of liver tissue of rats in each group was detected by Western blot at 12 and 24 hours after injection of adenovirus.(2) Liver function test of recipient rats after liver transplantation:liver functions of recipient rats [alanine transaminase (ALT),aspartate transaminase (AST),alkaline phosphatase (ALP),gamma-glutamyl transferase (GGT)] were detected at l,3,7 and 14 days postoperatively.(3) Pathological histology of liver tissue and injury scores of recipient rats in the 3 groups after liver transplantation:paraffin sections of recipient rats in the 3 groups at postoperative 1 and 14 days were stained by HE staining and observed by light microscope,and were evaluated by Suzuki damage score standard.(4) Relative expressions of HIF-1α,VEGF and HO-1 in liver tissue of recipient rats were detected by Western blot.(5) Von Willebrand factor (vWF) in liver tissue of recipient rats at 14 days postoperatively was detected by immunofluorescence staining and small vessels were counted.Measurement data with normal distribution were represented as x ±s.Comparison between groups was analyzed by the independent-sample t test,comparison among groups was done using one-way ANOVA,and pairwise comparison was analyzed by the LSD test.Results (1) Detection of transfection efficiency of adenovirus before operation:the relative expression of HO-1 of liver tissue of rats at 12 and 24 hours preoperatively after injection of adenovirus was 1.08±0.16 and 1.08±0.26 in the empty virus group,1.18±0.21 and 1.39±0.19 in the induced group,0.87±0.26 and 0.57±0.12 in the inhibited group,respectively,with statistically significant differences in different time points (F =4.232,36.513,P＜ 0.05).(2) Liver function test of recipient rats after liver transplantation:level of ALT at 3 days postoperatively in the empty virus group,induced group and inhibited group was (504±67)U/L,(438±47)U/L and (490±39)U/L,with a statistically significant difference (F=3.517,P＜0.05).Levels of ALT,AST and ALP at 7 days posto-peratively were (443±49) U/L,(430± 34) U/L,(455± 38) U/L in the empty virus group and (382± 49) U/L,(372±50) U/L,(394±25) U/L in the induced group and (493±44) U/L,(455±62) U/L,(470±72) U/L in the inhibited group,respectively,with statistically significant differences (F =10.950,5.667,5.398,P＜0.05).Levels of ALT,AST,ALP and GGT at 14 days postoperatively were (394±46)U/L,(361 ±68)U/L,(417 ±17)U/L,(4.5±1.1)U/L in the empty virus group and (283±47) U/L,(288±60) U/L,(332±46) U/L,(2.5±0.5) U/L in the induced group and (446± 43) U/L,(422± 51) U/L,(423± 63) U/L,(4.3 ± 1.3) U/L in the inhibited group,respectively,with statistically significant differences (F=26.906,9.924,8.013,9.279,P＜ 0.05).(3) Pathological histology of liver tissue and injury scores of recipient rats in the 3 groups after liver transplantation:liver cell swelling,loose cytoplasm and a varying quantity of inflammatory cell infiltration in the portal regions in the liver tissue of 3 groups were observed at 1 day postoperatively.A few inflammatory cell infiltrations in the portal regions,basically normal liver cell arrangement and a slightly swelling of liver cell were found in the empty virus group at 14 days postoperatively.Reduced liver cell swelling and basically normal structure of liver lobule were observed in the induced group.There were small patchy or focal necrosis of liver cell,masses of inflammatory cell infiltration in the portal regions and damage of bile duct in the inhibited group.Suzuki score at 1 day postoperatively in the empty virus group,induced group and inhibited group were respectively 6.7± 1.7,6.1 ± 1.2 and 7.6± 1.3,with no statistically significant difference (F=2.257,P＞0.05).Suzuki score at 14day postoperatively in the empty virus group,induced group and inhibited group were respectively 4.0±0.8,2.9± 0.8 and 5.1± 1.4,with a statistically significant difference (F=9.776,P＜0.05).(4) Western blot results:the relative expressions of HIF-1α and VEGF (43 KD) in liver tissue of recipient rats at 1 day postoperatively were 0.21±0.10,0.30±0.12 in the empty virus group and 0.23±0.09,0.34±0.14 in the induced group and 0.17± 0.06,0.29±0.11 in the inhibited group,respectively,with no statistically significant difference (F =0.902,0.410,P＞0.05).The relative expressions of VEGF (24 KD) and HO-1 in liver tissue of recipient rats at 1 day postoperatively were 1.21 ±0.25,0.55±0.12 in the empty virus group and 2.13±0.40,0.72±0.12 in the induced group and 0.91±0.22,0.26±0.07 in the inhibited group,respectively,with statistically significant differences (F=35.158,39.082,P ＜ 0.05).The relative expressions of HIF-1α,VEGF (43 KD),VEGF (24 KD) and HO-1 in liver tissue of recipient rats at 7 days postoperatively were 0.49±0.22,0.46±0.13,0.98± 0.37,0.98±0.37 in the empty virus group and 0.83±0.26,0.63±0.19,1.60±0.33,1.49±0.46 in the induced group and 0.24±0.09,0.30±0.12,0.64±0.18,0.75±0.26 in the inhibited group,respectively,with statistically significant differences (F=16.853,10.021,20.756,8.156,P＜0.05).(5) Immunofluorescence staining results:number of small vessels at 14 days postoperatively in the empty virus group,induced group and inhibited group was respectively 7.9±2.0,10.6± 1.9 and 7.6 ± 1.9,with a statistically significant difference (F=5.921,P＜0.05).Conclusion HO-1 could promote expressions of HIF-1α and VEGF in liver tissue after liver transplantation ischemia-reperfusion injury and regeneration of intrahepatic vascular plexus,and it also alleviate bile duct ischemia-reperfusion injury after liver transplantation.

Objective To explore the effectivity and feasibility of uindirectional barbed suture for primary closure of common bile duct on Laparoscopic Exploration of Common Bile Duct Stones, Methods From January 2013 to August 2015,109 cases of primary closure of common bile duct after Laparoscopic Exploration of Common Bile Duct Stones were performed in this hospital.The characteristics of these cases were retrospectively comparatively analyzed.Results Conventional braided Sutures in 68 cases(group A),barbed Suture Devices in 41 cases (group B).There was no difference in postoperative hospital stay,intraoperative blood loss and complication rate between the two groups.Differences in bile duct incision suture time and operative time between the two groups were significantly different.Conclusion The application of uindirectional barbed suture for primary closure of common bile duct using barbed suture after Laparoscopic Exploration is a safe and effective way after treatment of choledocholithiasis.This method cuold reduce the difficulty of operation,and shorten the operation time and the learning curve.

Objective To evaluate upfront common bile duct suturing against T-tube drainage after exploration in the treatment of common bile duct stone.Methods 253 cases of extrahepatic bile duct stones treated at our department from 2008 June to 2012 January were randomly divided into primary suture group and T tube drainage group,by t test or analysis of variance independent sample comparison.Results All operations were successful.Postoperative bile leakage was observed in 2 patients in group A and 4 in group B respectively(P ＞ 0.05),there was no reoperations in the two groups.In group B retrograde biliary tract infection developed in one and was cured by biliary tract flush combined with antibiotics administration.The abdominal drainage was bile tainted fluid about 5-10 ml a day in bile leakage cases in both groups and healed itself in 3-4 days without fever,jaundice symptoms.There was no other severe complications such as pancreatitis.The postoperative biliary complication rate (P =0.802),operative time (P =0.137),intraoperative blood loss (P =0.069) and liver function recovery(ALT P =0.087,AST P =0.752,TBIL P =0.459,DBIL P =0.217,ALP P =0.576,GGT P =0.362) was not significantly different between the two groups.In group A postoperative flatus passing (P =0.037),postoperative fluid volume (P =0.008),postoperative hospital stay(P =0.015) were better than that in T-tube drainage group.At 3 to 12 months follow-up,no patients were found to have residual stones and biliary stricture in group A and group B.Conclusions With the definite indication and proficient surgical technology,primary suture of common bile duct after exploration is a safe and effective way after treatment of choledocholithiasis.

Background and purpose: Because of the aggressive nature of hilar cholangiocarcinoma and the absence of effective adjuvant therapy, surgical radical resection offers hilar cholangiocarcinoma patients the only choice. Research focus include preoperative assessment, the use of preoperative biliary drainage, the range of hepatic resection, and the range of lymphadenectomy. To investigate the clinical experience and efifcacy of combined hepatectomy in the treatment of hilar cholangiocarcinoma. Methods: Two hundred and seven patients with hilar cholangiocarcinoma treated surgically in the First Afifliated Hospital of Kunming Medical University form Jan. 2007 to Oct. 2013 were retrospectively analyzed. Results:Of the 207 patients, 125 patients who received radical resection (R0 resection) and the curative resection rate was 60.4%. One hundred and iffty-six cases were treated in combined hepatectomy group, 51 cases in non-hepatectomy group, the rate of R0 resection was 70.5%in hepatectomy group and 29.4%in non-hepatectomy group, and the difference was signiifcant (P<0.01). Two patients died perioperatively, the main postoperative complications included hepatic function insufifciency and bile leakage. One hundred and seventy-two patients were followed up, the median survival time of the 102 patients who received R0 resection was 45 months, and the 1, 3, 5 year survival rates were 96.1%, 59.1%and 17.2%. The median survival time of the 70 patients who received R1-2 resection was 26 months, and the 1, 3 year survival rates were 81.3%and 19.2%, and none of the patient survived for over 5 years. The survival rate of patients who received R0 resection was signiifcantly higher than those who received R1-2 resection (χ2=39.121, P<0.01). In the hepatectomy group was awarded the R0 resection in patients with postoperative 1, 3, 5 year survival rate was 97.8%, 63.9% and 18.0%, in non-hepatectomy group received R0 resection in patients with postoperative 1, 3, 5 year survival rate was 83.3%, 20.8%and 8.3%. There were signiifcant differences in the postoperative survival rate between both group (χ2=5.988, P=0.014). Conclusion:Radical excision is the key to improve the long term survival. Combined hemihepatectomy and standardized lymph node resection has signiifcantly improved the radical resection rate and the efifcacy of treatment for hilar cholangiocarcinoma.

Objective:To investigate the effects of heme oxygenase-1 (HO-1) on hepatic sinusoidal endothelial cells (LSECs) proliferation, migration, and hepatocyte proliferation.Methods:Eighteen male C57BL/6 mouse aged 6-8 weeks old were underwent partial hepatectomy. Cell proliferation and HO-1 expression in residual liver tissue were detected by immunofluorescence histochemistry at 0 d, 2 d and 4 d after operation. In vitro, LSECs were transfected with adenovirus carrying HO-1 gene (HO-1 group), and the cells were transfected with empty vector adenovirus and the non-transfected cells were used as control. In addition, LSECs from different transfection groups were co-cultured with hepatocyte without contact to evaluate the effect of HO-1 expression on promoting hepatocyte proliferation. Western Blotting and RT-PCR were used to detect the protein and mRNA expression of HO-1, inhibitor of DNA binding and or differentiation (Id1), hepatocyte growth factor (HGF) and Wnt2. Cell proliferation was detected by EdU. The ability of cell migration was detected by Transwell migration assay.Results:Compared with 0 d after hepatectomy, LSECs proliferation and HO-1 expression within LSECs were increased significantly at 4 d after surgery. EdU positive rate of LSECs in HO-1 group (27.20±4.80)% was higher than that in empty vector group (12.47±3.30)% and non-transfected group (15.97±2.50)%. The number of LSECs migration in HO-1 group (258.70±36.56) was higher than that in empty vector group (122.00±38.16) and non-transfected group (107.70±30.01). The protein and mRNA expression level of HO-1, Id1, HGF and Wnt2 in HO-1 group were higher than that in empty vector group and non-transfected group. EdU positive rate of hepatocytes that co-cultured with LSECs in HO-1 group (18.33±2.52) % was higher than that in empty vector group (11.33±1.53)% and non-transfected group (11.7±2.08)%. The differences were statistically significant (all P<0.05). Conclusion:Up-regulation of HO-1 promoted LSECs proliferation and migration of, as well as up-regulation of HO-1 in LSECs enhanced the capacity of LSECs to promote hepatocyte proliferation.

Objective To investigate the protective effect of gastrodin (Gas) on the hepatic ischemia-reperfusion injury (HIRI) in mice,and study possible mechanisms.Methods Forty male C57BL/6 mice were randomly divided into sham-operated group (Sham),HIRI group,gastrodin-treated groups with low and high does (Gas-L,Gas-H,respectively),and cobalt protoporphyrin(CoPP) group(n =8).Mice in Gas-L,Gas-H,and CoPP groups were injected intraperitoneally with individual drugs and dose before operation.Except for Sham group,an 70％ volume HIRI model was established by means of 60 minutes ischemia and then 6 hours reperfusion in the other groups.The levels of serum AST and ALT in each group were compared.The levels of superoxide dismutase (SOD) and malondialdehyde (MDA) in the liver tissue were measured;The level of TNF-αt,IL-6,IL-10 and heme oxygenase-I (HO-1) mRNA level were detected by real time quantitative PCR.Liver tissue was stained with H&E.The hepatocyte apoptosis was studied by TUNEL.Results Respectively,the serum ALT level in the HIRI,Gas-L,Gas-H and CoPP groups was (5 057.34±290.80)U/L,(3 917.49±198.10) U/L,(3 645.63± 171.10) U/L,(2 977.78± 179.00) U/L,and the ALT level was (5 871.25 ± 819.01) U/L,(4 660.88 ± 505.96) U/L,(4 182.00 ± 507.51) U/L,(3 788.65±462.14)U/L.The serum level of ALT and AST in Gas-L,Gas-H and CoPP groups was lower than HIRI group (P＜ 0.05).The apoptotic index in Gas-L,Gas-H and CoPP groups respectively was (37.89±4.27)％,(32.59±3.78)％,(20.45-±2.49)％,which was lower than group (58.92±3.32)％ (P＜ 0.05).Compared with HIRI group,the TNF-α and IL-6 mRNA level in Gas-L,Gas-H and CoPP groups were decreased,and the levels of IL-10 and HO-1 mRNA were increased.Simultaneously,the activity of SOD was promoted,whereas the levels of MDA was reduced (P＜0.05).Conclusions Gastrodin shows an protective function aganist liver ischemia-reperfusion injury in mice by inhibiting inflammation,oxidative stress and cell apoptosis.The up-regulation of HO-1 by gastrodin may be the possible mechanism.