Objective To investigate the effect of exosomes derived from hepatocellular carcinoma cells on the polarization of tumor-associated macrophages (TAMs), and to reveal the novel mechanism of hepatocellular carcinoma formation. Methods Hepatocellular carcinoma cell-derived exosomes were isolated by ultracentrifugation, and the characteristics of exosomes were identified by transmission electron microscope (TEM), Dynamic Light Scattering (DLS), and Western blotting. The model of macrophage polarization was induced and verified by quantitative real-time PCR and Western blotting. The t -test was used for comparison of normally distributed continuous data between two groups. A one-way analysis of variance was used for comparison between multiple groups, and the LSD- t -test was used for further comparison between two groups. Results TEM showed that hepatocellular carcinoma cell-derived exosomes were round or oval vesicles, LDS showed that the exosomes had a particle size of 172.65±2.34 nm, and Western blotting showed highly positive expression of the biomarkers TSG101 and CD63 in exosomes. There was a significant increase in the expression of CD68 after the addition of 15 ng phorbol ester to induce human-derived mononuclear macrophages for 24 hours to achieve adherent growth (1.00±0.25 vs 6.67±0.98, t =11.20, P < 0.001). Western blotting showed that compared with the control group (L02 cell-derived exosomes), the hepatocellular carcinoma cell-derived exosomes (at low, middle, and high doses) induced M2 polarization of macrophages and increased the expression of the markers Arg-1 and CD163 (all P < 0.05). Conclusion Hepatocellular carcinoma cell-derived exosomes promote M2 polarization of TAMs.

Objective To evaluate the association of the size of thyroid nodules and accuracy of fineneedle aspiration cytology in diagnose of thyroid nodules.Methods 691 thyroid nodules in 630 patients pathologically confirmed were retrospectively analyzed in our hospital.All imaging data of preoperative ultrasound-guided FNAC were collected in our review.Yields of FNAC were divided into six levels according to the classification criteria of the Bethesda system(level Ⅰ,insufficient material or nondiagnosed;level Ⅱ,benign ; level Ⅲ,atypical hyperplasia; level Ⅳ,follicular neoplasm ; level Ⅴ,suspicious for malignancy; level Ⅵ,malignant),＞level Ⅳ was the malignant cytologic criteria for diagnosis of thyroid nodules.According to the maximal diameter of thyroid nodules,the nodules were divided into group A(L≤0.5 cm),group B(0.5 cm＜L＜1.0 cm) and group C(L≥1.0 cm).Postoperative pathologic results were taken as the gold standard.Results Of 691 nodules,there were 176(25.47％),298(43.13％) and 217(31.40％) in group A,group B and group C respectively.Among the three groups,accuracy of ultrasound-guided FNAC in group B (90.94 ％) was higher than in group A(80.11％) and group C(83.41 ％),with statistically significant(P ＜0.05).There was not statistically different between group A and group C(P ＞0.05).The specificity,positive predictive value and negative predictive value were not statistically different among three groups(P ＞0.05).Conclusions The size of thyroid nodules was partly associated with accuracy of ultrasound-guided FNAC.

Objective To investigate the effect of a modified cuff insertion technique named “portal vein branch-sling” (PBS) method for portal vein (PV) anastomoses in rat liver transplantation.MethodsA total of 60 cases of orthotopic liver transplantation (OLT) using 2-cuff technique in rats were performed in our transplantation center.The PBS technique and conventional methods were used respectively in study and control groups.Time for PV anastomoses,anhepatic phase (AP) and surgery of recipients,successful rate for PV cuff insertion and operation were statistically compared between the different groups.Results35 cases of OLT using PBS technique in rats were assessed.The successful rate of PV cuff insertion was 94.3％,and the time of PV anastomoses using the modified PBS cuff insertion technique was only 1.9 ± 0.7min.The successful rate of operation was 80％with a 21.8±2.2min AP.The study group showed significant differences (P＜0.05) compared with conventional group.The survival rate in the first week was 85.8％ with no statistical difference (P＞0.05).ConclusionsThe modified PBS cuff insertion technique for PV anastomoses of OLT has advantages in speed,safety and stabilization,and should be considered as one of the best available PV anastomosis technique for OLT in rats.

ObjectiveTo identify the proteins presented variation during ischemia/reperfusion injury (I/RI) in liver transplantation (LT).MethodsAt Mar.2009,a total of nine liver sample tissues,three samples obtained from each donor liver at three different time points,were studied.The time points were as follows:(1) the time point immediately after donor liver was harvested (T1) ; (2) the time point just before the anastomosis of hepatic artery and potal vein (T2),during this time period the donor liver was kept in ice cold and transported,then back-table prepared,during time from T1 to T2,ischemia injury occurred; (3) the time after donor reconnected to the recipient (T3),during which the donor liver was subjected to reperfusion injury.The three time points represent normal control,ischemia injury,and reperfusion injury respectively.By using method of 2DE-MALDI-TOF and mass-spectrum,the differential expression of these protein at T1,T2,and T3 time point were compared.ResultsFrom the 1580 proteins that were isolated,19 ischemia/reperfusion injury (I/RI)-related proteins that varied markedly from time point T1 to T2 and from time point T2 to T3 were identified.They were metabolic enzyme,molecular chaperone,redoxase,cytoskeleton protein,signal transduction protein,and binding protein,respectively.The character of each protein was further analized based on their functions.ConclusionsFor the first time,we detected the protein related to I/RI during LT.We obtained independent proteomics information of ischemia injury,and reperfusion injury,respectively.Further functional analyses of these proteins may improve the understanding of the critical biological processes during I/RI.Medicine intervention on the I/RI-related proteins may result in improved survival of the recipients.

Objective To evaluate the efficacy and indications of salvage liver transplantation for patients with recurrent hepatecellular carcinoma(HCC)after liver resection.MethodsThe clinical data of 35 HCC patients who received salvage liver transplantation after liver resection at the Third Affiliated Hospital of Sun Yatsen University from October 2003 to March 2006 were retrospectively analyzed.All patients were divided into the salvage liver transplantation(SLT)group(n = 19)and extended SLT group(n = 16).Perioperative condition,postoperative complications and prognosis of the 2 groups were compared.The survival rate was analyzed and compared by the Kaplan-Meier method and Log-rank test,respectively.Results The anhepatic phase,ischaemic time,operation time,intraoperative blood loss,packed red blood cell transfusion,fresh frozen plasms transfusion,mobidity and retransplantation rate were(32 ± 9)minutes,(8.0 ± 2.1)hours,(7.6 ± 1.5)hours,2300 ml,8 U,23 U,6/19 and 2/19 in the SLT group,and(34 ± 7)minutes,(7.4 ± 2.3)hours,(7.4 ± 2.0)hours,2750 ml,12 U,20 U,4/16,1/16 in the extended SLT group,respectively,with no significant difference between the 2 groups(t=0.726,-0.804,-0.366,Z=-0.348,-0.549,-0.149,x2 =0.184,0.203,P＞0.05).The perioperative mortality,tumor recurrence rate were 0 and 2/19 in the SLT group,and 4/16 and 9/16 in the extended SLT group,with significant differences between the 2 groups(x2 = 5.363,8.426,P ＜ 0.05).The 1-,3-,5-year cumulative survival rates were 100%,84% and 84% in the SLT group,and 75%,33% and 33% in the extended SLT group.The 1-,3-,5-year tumor-free survival rates were 100%,89% and 89% in the SLT group,and 48%,29% and 19% in the extended SLT group.There were significant differences in the cumulative and tumor-free survival rates between the2 groups(x2 =11.58,19.31,P＜0.05).Conclusions The efficacy of SLT is satisfactory in the treatment of recurrent HCC.The optimal indication for SLT is Milan criteria.

Objective To study the related factors associated with the reversal of posttransplant diabetes mellitus (PTDM) following liver transplantation. Methods The clinical data of 62patients with PTDM in 232 patients receiving liver transplantation (26. 7 %) were retrospectively analyzed and the patients were divided into two groups: patients with transient PTDM (34 cases) and those with persistent PTDM (28 cases). Pre-operative and post-operative variables, including sex,age, body mass index, family history of diabetes, hepatitis B virus infection, pretransplantation fasting plasma glucose, the immunosuppressant regime, FK506 concentration and duration of steroid usage, were analyzed retrospectively. Results The variables, including sex, age, body mass index,family history of diabetes, hepatitis B virus infection, pretransplantation fasting plasma glucose,FK506 concentration at month 1, 3 and 6 after operation, rate of cyclosporine usage and duration of steroid usage had no significant difference between the two groups (P＞0. 05). Compared with the persistent PTDM patients, the transient PTDM patients were characterized by younger age at the time of transplantation (54 ± 8 vs. 42 ± 6 years, P＜0. 05), longer time before the development of PTDM (18 ± 23 vs. 35 ± 42 days, P＜0. 05), and higher rate of mycophenolate mofetil or sirolimus usage (0vs. 8. 9 %, P＜0. 05). Based on a multivariate analysis, age at the time of transplantation was determined as the single independent predictive factor associated with reversal of PTDM following liver transplantation (odds ratio: 1. 312, 95 % confidence interval: 1. 005 - 1. 743). Conclusion Age at the time of transplantation, duration before the development of PTDM and rate of mycophenolate mofetil or sirolimus usage are associated with reversal of PTDM following liver transplantation. Among these factors, age at the time of transplantation is only the single independent predictive factor.

Objective To evaluate the effect of liver transplantation for end-stage autoimmune liver disease (ESALD) and summarize the clinical experience of liver transplantation in the treatment of ESALD.Methods The clinical data of 11 ESALD cases who underwent liver transplantation from September 2003 to July 2009 were analyzed retrospectively. There were 2 males and 9 females ( median age, 44. 2 ± 8. 7years). The indication of liver transplantation was end stage of primary biliary cirrhrosis (8 cases),autoimmune hepatitis (2 cases), and primary sclerosing cholangitis ( 1 case). In all cases, modified piggyback liver transplantation with venacavaplasty was carried out. Postoperatively all patients were treated with immunosuppressive agents including tacrolimus (or cyclosporine A) and prednisone, some patients were treated additionally with mycophenolate mofetil and ursodeoxycholic acid. Results Postoperatively 2patients of primary biliary cirrhosis died, one of lung infection and multiple organ failure on the 5th postoperative day, the other dying of sepsis and graft dysfunction on the 964th postoperative day. Five cases suffered from episodes of acute cellular rejection within 1 month after transplantation and was successfully reversed by strengthened immunosuppressive therapy. Nine patients recovered satisfactorily and with excellent life quality until now. Patients were followed up from 7 months to 62 months with the median follow-up time of 38 months. The recipient survival rate at 1 year and 3 years was 91% and 82% ,respectively. One patient has now survived for 5 years. No recurrent ALD case was found during follow up.Conclusions Orthotopic liver transplantation is an exclusive treatment for ESALD. Optimum operation timing and effective immunosuppressive treatment are very important for decreasing occurrence of complications.

Objective To investigate the diagnosis and interventional therapeutic technology for the obstruction of hepatic vein(HV)or inferior vena cava(IVC)after liver transplantation.Methods In the 831 patients who received orthotopic liver transplantation(OLT)and 26 patients who received living donor liver transplantation(LDLT),11 cases were confirmed with HV or IVC obstruction by venography and received interventional treatment from 2 to 111 days after liver transplantation.Of the 11 patients,five had the obstruction of HV anastomosis,five had the obstruction of IVC anastomosis,and one had the obstruction of HV and IVC anastomosis.In the eleven patients,five patients underwent OLT,four patients underwent LDLT,and two pediatric patients underwent reduced-size OLT.Before interventional treatment,9 patients received CT enhanced scans,2 received MR enhanced scans.Follow-up evaluations included liver or renal function tests,clinical symptom,and monitoring of HV or IVC flow.Pressure gradients before and after interventional treatment were compared by using a paired t test.The imaging data and interventional therapeutic technology of 11 cases were retrospectively analyzed.Results In all 11 patients,CT or MRI could clearly show congested areas of the liver,and the location and degree of HV or IVC obstruction.Of the 11 patients,four with HV obstruction and five with IVC obstruction were treated with stent placement,one with HV obstruction was treated with percutaneous transluminal angioplasty(PTA),one with HV and IVC obstruction was treated with HV PTA and IVC stent placement.Interventional technical success was achieved in all patients.The venous pressure gradient across obstruction was significantly reduced from(16.5±4.1)mm Hg(1 mm Hg=0.133 kPa)before the procedure to(2.9±1.7)mm Hg after the rocedure(t= 11.5,P＜0.01).Clinical improvement was noted in 10 patients except one pediatric patient who died of multiple-organs failure at the 9 th day after the treatment During the follow-up period of 9 to 672 days,two patients with PTA treatment had recurrent HV stenosis within one month after treatment,no patient with stent placement developed venous restenosis.No major complications occurred during the procedures.Conclusions Stent placement is safe and effective for HV or IVC obstruction after liver transplantation.CT or MRI before treatment is of important value for the diagnosis of congested areas of the liver,and the observation of HV or IVC obstruction.

Objective To investigate the indications, complications and survival results of combined liver-kidney transplantation. Methods From Oct 2003 to Dec 2008, the clinical data of 13 patients who underwent combined liver-kidney transplantation (CLKTs) were retrosptiverly analyzed in our institution. The perioperative mortality rate, complications and the result of follow-up were analyzed.Results The perioperative mortality rate (within 30 days) was 30.8% (4/13). Postoperative complications included intrabdominal bleeding in 4 patients ( 30. 8% ); pulmonary infection in 7 patients (53.8%); acute renal rejection in one (7. 7% ). Survivors were followed up from 4.4 to 60 months, with the median time of 40 months. Eight patients have survived more than 1 year; six patients have survived more than 2 years; five of them have survived for more than 3 years; and three of them have survived for more than 4 years, with one surviving for more than 5 years. One patient had undergone liver transplantation ( case 2 ) and two patients had had kidney transplantations ( case 3 and case 4 ) before this CLKTs.Postoperatively case 4 died of pulmonary infection and multiple organ failure at day 29, while case 2 and case 4 survived respectively 40 m, 48 m after CLKTs. Conclusions CLKTs is an effective therapy for end-stage liver and kidney disease. CLKTs for patients with irreversible liver and renal insufficiency after initial liver transplantation or kidney transplantation was feasible.

Objective To determine the timing for therapy and efficacy for different types of hepatic artery stenosis (HAS) after orthotopic liver transplantation(OLT). Methods From October 2003 to May 2007, a total of 21 patients had hepatic artery stenosis after OLT in this hospital. Of the 21 patients, 19 underwent stent placement in their narrowed hepatic arteries and 2 were regularly followed up. Liver function, clinical outcomes, and the hepatic artery potency were reviewed. Results The occurring rate of HAS was 3.43% (21/613) and its median time of diagnosis was 146 days (range, 2-515 days). Six patients with early HAS were treated with interventions and 2 of them died.For the 4 surviving patients, 2 received retransplantation. For the 15 patients with late HAS, 13 were treated with interventions and 4 of them died. Two patients received retransplantation. Seven surviving patients had abnormality in liver function. Another 2 patients had normal liver function because of hepatic portal form compensatory circulation. Conclusion The presence of ischemic bile duct lesion and whether or not favourable compensatory circulation exists or not should be considered before individualized therapeutic regimens adopted according to postoperative HAS types.