Objective To analyze the related immune parameters in liver transplantation recipients to help guide adjusting future immunosuppressants.Methods Fifty-three tolerant recipients after liver transplantation were included in this study.They had normal liver functions and took only one immunosuppressive drug.They were divided into the 3 ～5 years tolerance group (3Y group,n =21) (including 5 years) after transplantation,and the 5 ～ 11 years tolerance group (5Y group,n =32).Another 15 liver transplantation recipients who had twice or more recurrent acute rejections were enrolled as the RJ group and 32 age-matched healthy donors served as the HC group.The natural killer (NK) cells,B cells,memory T cells,regulatory T cells (Tregs) and the subsets were detected by flow cytometry.The plasma TGF-β level was evaluated using ELISA.Results The percentages of NK cells (3Y group 19.00 ± 0.09,5Y group 20.00 ±0.09,HC group 14.00 ±0.07,RJ group 9.00 ±0.03) and total Tregs within the CD4+ cells (3Y group 6.26 ±2.33,5Y group 4.80 ±2.21,HC group 3.04 ± 1.25,RJ group 1.09 ±0.81) in the 3Y,5Y and HC groups were significantly higher than those in the RJ group (P ＜ 0.05).The proportions of resting Tregs (3Y group 0.23 ±0.07,5Y group 0.16 ±0.02,HC group 0.07 ±0.02,RJ group 0.05 ±0.01),active Tregs (3Y group 0.56 ± 0.11,5Y group 0.42 ± 0.15,HC group 0.08 ± 0.02,RJ group 0.05 ± 0.01) and non-suppressive Tregs (3Y group 3.51 ±0.80,5Y group 3.71 ±0.41,HC group 1.44 ±0.14,RJ group 2.15 ± 0.62) increased significantly in the tolerant recipients after liver transplantation when compared with those in the HC and RJ groups (all P ＜ 0.05).No significant differences on B cells,memory T cells and TGF-β level were detected among in four groups.Conclusions The NK cells,total Tregs,resting Tregs,active Tregs and non-suppressive Tregs increased significantly in the tolerant recipients after liver transplantation.These might serve as potential biomarkers for immune tolerance.

Objective To estimate the safety,feasibility and generalization of three point single-incision laparoscopic cholecystectomy (SILC).Methods The clinical data of 1 126 patients who underwent three-point SILC at the second department of Hepatobiliary Surgery of Zhu Jiang Hospital,Southern Medical University From January 1,2011 to December 30,2015 was analyzed retrospectively.The patient who were indicated for conventional laparoscopic cholecystectomy were included,but those suspected malignant diseases of gallbladder were excluded.Results Of the 1126 patients,the surgery was performed successfully in 923 patients,and 192 patients need extra ports due to the adhesion and difficulty of exposing the Calots triangle,and 11 were converted to open surgery due to severe adhesion,with the success rate being 81.9％.The operating time was (29.5 ± 12.2) min (from the entrance of laparoscope to the removing of gallbladder),the blood loss was (8.7 ± 7) ml,and the hospital stay time was (1.4 ± 0.7) d (after surgery).There were three cases of bile duct injury:two of them were bile leak of aberrant duct,one of them was bile leak of cystic duct damaged by heat.And there was one case of injury of duodenum,22 cases of umbilicus hematoma,13 cases of hematoma of thorax,and 2 cases of thoracic hemorrhage who required surgery.There were no hernia,aerothorax and so on.Conclusion Three point SILC is a technology that is safe,maneuverable and suitable for being carried out in clinical practice.

OBJECTIVETo determine whether the miRNA expression profile in peripheral blood mononuclear cells (PBMCs) differs between liver transplant recipients with long-term stable survival and those with acute rejection.

METHODSTwenty-nine liver transplant recipients with long-term stable survival (STA) group, 10 recipients with acute rejection (RJ group), and 17 healthy subjects (control group) were recruited for genome-wide microarray analysis of miRNA expressions in the PBMCs. The differentially expressed miRNAs among the 3 groups were validated by real-time PCR, and the targets of these miRNAs were predicted.

RESULTSCompared with the RJ group, the STA group showed down-regulation of 13 miRNAs in the PBMCs. Of these down-regulated miRNAs, miRNA-18b, miRNA-340 and miRNA-106b were validated by real-time PCR, and the latter two miRNAs were predicted to target the TGF-β pathway.

CONCLUSIONSThe differentially expressed miRNAs in liver transplant recipients with long-term stable survival, namely miRNA-18b, miRNA-340 and miRNA-106b, can be potential clinical biomarkers to predict the outcomes of liver transplantation.

Biomarkers ; metabolism ; Case-Control Studies ; Down-Regulation ; Graft Rejection ; Humans ; Leukocytes, Mononuclear ; metabolism ; Liver Transplantation ; MicroRNAs ; metabolism ; Oligonucleotide Array Sequence Analysis ; Real-Time Polymerase Chain Reaction ; Survival Rate ; Transforming Growth Factor beta

OBJECTIVETo detect the expression of Nusap1 of surgical margins in hepatocellular carcinoma (HCC) and investigate its association with early tumor recurrence.

METHODSThe expression of Nusap1 in the surgical margins of HCC, which were histopathologically negative for tumor cells, was examined using immunohistochemistry in 61 HCC cases.

RESULTSFifteen of 21 (71.4%) cases with immunohistochemical positivity for Nusap1 expression in the surgical margins had early recurrence of HCC, a rate significantly higher than that in patients with negative Nusap1 expression (12/40, 30%) (P<0.05).

CONCLUSIONSNusap1 expression in the surgical margins of HCC is closely correlated to early postoperative recurrence and can serve as an indicator for predicting early recurrence of HCC.

Adolescent ; Adult ; Aged ; Carcinoma, Hepatocellular ; metabolism ; pathology ; surgery ; Female ; Humans ; Immunohistochemistry ; Liver Neoplasms ; metabolism ; pathology ; surgery ; Male ; Microtubule-Associated Proteins ; metabolism ; Middle Aged ; Neoplasm Recurrence, Local ; pathology ; Young Adult

OBJECTIVETo study the expression of cellular inhibitor of apoptosis protein-2 (C-IAP2) mRNA and protein in hepatocellular carcinoma (HCC) and its relationship with the clinical outcomes.

METHODSQuantitative PCR and immunohistochemical staining were used to detect the expression of C-IAP2 mRNA and protein in the tumor tissues and corresponding adjacent non-cancerous tissues from HCC patients.

RESULTSThe expression of C-IAP2 mRNA in HCC tissues was 2.70 folds higher than that in the non-cancerous tissues (P<0.001). The expression rate of C-IAP2 protein in HCC tissues was 70.8%, significantly higher than that in the non-cancerous tissues (27.8%, P=0.001). The expression of C-IAP2 mRNA and protein was associated with the tumor emboli, lymph node metastasis, AFP level, histological differentiation, TNM stage, postoperative recurrence and metastasis (P<0.05), but not with the patients' gender, age, HbsAg positivity, number of tumors, cirrhosis or the presence of tumor encapsulation (P>0.05).

CONCLUSIONThe expression of C-IAP2 in HCC is associated with tumor recurrence and metastasis, and can be a biological marker for prognostic evaluation of HCC.

Baculoviral IAP Repeat-Containing 3 Protein ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; Liver Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Prognosis ; Ubiquitin-Protein Ligases

OBJECTIVETo study the expression of special AT-rich sequence binding protein 1 (SATB1) in hepatocellular carcinoma (HCC) cell lines with different invasive capacities.

METHODSSATB1 expression was detected using real-time fluorescence quantitative PCR, RT-PCR, Western blotting and immunofluorescence in immortalized liver cell line HL-7702, noninvasive HCC cell lines HepG2 and SMMC-7721, MHCC97L cells with low invasiveness, and highly invasive cell lines MHCC97H and HCCLM3.

RESULTSIn comparison with HL-7702 cells, all the 5 HCC cell lines showed overexpression of SATB1 mRNA, which was the highest in the highly invasive HCCLM3 and MHCC97H cells, followed by MHCC97L cell line, and then by SMMC-7721 and HepG2 cell lines (P<0.001). The relative expression quantity of SATB1 protein in HepG2, SMMC-7721, MHCC97L, MHCC97H, and HCCLM3 cell lines was 0.271±0.002, 0.351±0.023, 0.621±0.026, 0.878±0.026, and 1.236±0.006, respectively. SATB1 expression level in HCCLM3 cell line was 4.6-fold higher than that in HepG2 cell line (P<0.001). SATB1 was found to localize in the cytoplasm and cell nuclei of the 5 HCC cell lines, and the highly invasive HCCLM3 and MHCC97H cell lines showed a strong positive staining for SATB1 in immunofluorescence assay.

CONCLUSIONSATB1 expression levels differ distinctly between the HCC cell lines with different invasive capacities and are possibly associated with the metastatic potentials of the cells.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Matrix Attachment Region Binding Proteins ; metabolism ; Neoplasm Invasiveness ; RNA, Messenger ; genetics

OBJECTIVETo investigate the surgical techniques and appropriate perioperative management for ensuring successful orthotopic liver transplantation (ROLT) in rats.

METHODSBased on the double-cuff technique of Kamada, we modified the surgical techniques of separation, perfusion and cold preservation of the donor liver, shearing and anastomosis of the suprahepatic vena cava with optimized postoperative infusion protocols and animal care.

RESULTSTwo hundred and seventy rats underwent ROLT and a learning curve of the success rate was built to reflect the improvement of techniques. The learning curve showed steep improvements over the exploration stage, breakthrough stage and maturation stage, and the success rates increased sharply over time (0%, 71.1%, and 94.5%, respectively) until finally reaching over 90%. The shearing and anastomosis of the suprahepatic vena cava remained the most critical and difficult techniques in ROLT modeling.

CONCLUSIONProficient microsurgical techniques and meticulous nursing can reduce postoperative complications, enhance operational success rate and extend the survival time after ROLT.

Anastomosis, Surgical ; Animals ; Disease Models, Animal ; Graft Survival ; Liver ; surgery ; Liver Transplantation ; methods ; mortality ; Perioperative Care ; Rats ; Rats, Sprague-Dawley

OBJECTIVE： To study the effects of berberine hydrochloride on the pharmacokinetics of tacrolimus in rats after single or multiple administration， and to provide reference for clinical combination therapy. METHODS： 30 rats were randomly divided into 5 groups， with 6 rats in each group： group one was treated with single administration of tacrolimus； group two was treated with tacrolimus intragastrically， twice a day， for consecutive 1 week； group three was treated with single administration of berberine hydrochloride， 5 min later given single administration of tacrolimus； group four was treated with tacrolimus intragastrically， twice a day， for consecutive 1 week， and then given tacrolimus intragastrically once 5 min after intragastric administration of berberine hydrochloride on the 8th day； group five was treated with berberine hydrochloride intragastrically， twice a day， and given tacrolimus intragastrically every 5 min， for consecutive 8 d. The doses of berberine hydrochloride and tacrolimus were 200 mg/kg and 0.945 mg/kg. The blood samples 0.3 mL were collected from posterior orbital venous plexus of rats 0， 5， 15， 30 min and 1， 2， 3， 4， 6， 8， 12 h after last intragastric administration of tacrolimus. The concentration of tacrolimus in rat whole blood was determined by LC-MS/MS. DAS 2.0 software was used for pharmacokinetic study. RESULTS： Compared with group one， the pharmacokinetic parameters AUC0-12 h， AUC0-∞ and MRT0-12 h of tacrolimus in rats were decreased significantly in group three （P＜0.05），while there was no statistical significance in all pharmacokinetic parameters of tacrolimus in group four （P＞0.05）. Compared with group two， AUC0-12 h of tacrolimus was decreased significantly while CLz was increased significantly in group four （P＜0.05）； there was no statistical significance in all pharmacokinetic parameters of tacrolimus in group five （P＞0.05）. CONCLUSIONS： Single and multiple intragastric administration of berberine hydrochloride has a certain effect on the pharmacokinetics of tacrolimus in rats， it shows that there is a downward trend in blood drug concentration and needs to be used with caution.