Living related liver transplantation (LRLT) has been developed as a rescue for the ever deteriorating shortage of cadaveric donor liver, especially in Asian countries and areas where the concept of brain death has not widely been accepted. Inclusion criteria for the biologically suitable potential donor of LRLT should be strict and approved by the ethical committee of the hospital before clinical evaluation is taken. Anatomical, physiological and clinical practices have proved that donor mortality is acceptably very low. The early and middle-long term results for the recipient of LRLT is comparable with and even better than those of cadaveric liver transplantation. In mainland China, interest in LRLT is surging with the volume ever increased and donor's safety guaranteed.
Humans ; Liver Transplantation ; Living Donors

NY-ESO-1 is an important member of cancer-testis antigen family and is widely distributed among many cancer types. As a tumor-specific antigen with the strongest immunogenicity so far identified, it can induce spontaneous antibody and T-cell responses in patients with NY-ESO-1-positive tumors. Therefore, it has been a good vaccine candidate in the immunotherapy against many malignancies. This article reviews the recent research advances in NY-ESO-1 and its relevant vaccines.
Antigens, Neoplasm ; genetics ; immunology ; therapeutic use ; Cancer Vaccines ; immunology ; therapeutic use ; Clinical Trials as Topic ; Humans ; Immunotherapy ; Membrane Proteins ; genetics ; immunology ; therapeutic use ; Neoplasms ; genetics ; immunology ; therapy

Abdomen/surgery* ; Humans ; Laparoscopy/methods* ; Pelvic Floor/surgery* ; Perineum/surgery* ; Proctectomy ; Rectal Neoplasms/surgery*

OBJECTIVETo construct dendritomas by fusion of human dendritic cells with HLE cells, a human hepatocellular carcinoma cell line.

METHODSHLE cells were cultured in RPMI 1640 with 15% FCS. Human dendritic cells (DCs) were obtained from peripheral blood monocytes cultured in the presence of GM-CSF and IL-4 for 7 days, matured with TNF-alpha and PGE(2) for 2 days. The DCs and HLE cells were labeled with green fluorescence dye PKH67-GL and red fluorescence dye PKH26-GL, respectively, and fused in 50% polyethylene glycol (PEG) + 10% dimethyl sulfoxide (DMSO) to generate dendritomas for rapid fluorescence-activated cell sorting (FACS).

RESULTSDendritomas with dual red-green fluorescence were constructed successfully, and FACS analysis showed the effective fusion rate was 16.8%.

CONCLUSIONWith fluorescence dyes PKH67-GL and PKH26-GL as fusion markers, dendritomas for rapid fluorescence-activated cell sorting are constructed, which may throw new light on immunotherapy of hepatocellular carcinoma.

Cancer Vaccines ; biosynthesis ; Carcinoma, Hepatocellular ; pathology ; Cell Fusion ; methods ; Cell Line, Tumor ; Cells, Cultured ; Dendritic Cells ; cytology ; Humans ; Hybrid Cells ; Liver Neoplasms ; pathology

OBJECTIVETo investigate the protective effects of Shenfu Injection (SI) on hepatic ischemia-reperfusion injury in rats.

METHODSPartial liver ischemia-reperfusion model under room temperature was established in 60 rats, which were divided into the control group and the treated group randomly and each group was again classified into 3 subgroups with 30 min, 60 min and 90 min hepatic ischemia time rspectively. Rats in the treated group were injected with SI 10 ml/kg every day, while the control group treated with normal saline. Survival rate after 1 week was observed, the serum levels of aspartate aminotransferase (AST), malondialde hyde (MDA), surperoxide dismutase (SOD), tumor nicrosis factor alpha (TNF-a) and endothelin (ET) were detected, and hepatic biopsy was performed with light and electronic microscope.

RESULTSThe survival rate in the treated subgroup with 90 min' ischemia after 1 week was 90%, higher than that in the control subgroup significantly (P <0. 05), which was 60% ; and serum levels of AST, MDA, TNF-alpha and ET were lower and SOD was higher significantly (all P <0.05), as well as the degenerative and necrotic degree of hepatocyte and sinusoidal endothelial cells was lighter in the 3 treated subgroups, compared with the control group.

CONCLUSIONShenfu injection can eliminate oxygen free radical during hepatic ishemia-reperfusion so as to has a protective effect and attenuate hepatic ischemia reperfusion injury.

Animals ; Aspartate Aminotransferases ; blood ; Drugs, Chinese Herbal ; therapeutic use ; Endothelins ; blood ; Injections ; Liver ; blood supply ; Liver Diseases ; blood ; drug therapy ; prevention & control ; Malondialdehyde ; blood ; Protective Agents ; therapeutic use ; Rats ; Reperfusion Injury ; blood ; drug therapy ; prevention & control ; Superoxide Dismutase ; blood ; Treatment Outcome ; Tumor Necrosis Factor-alpha ; blood

BACKGROUNDOverexpression of G-protein coupled receptor 34 (GPR34) affects the progression and prognosis of human gastric adenocarcinoma, however, the role of GPR34 in gastric cancer development and progression has not been well-determined. The current study aimed to investigate the effect of GPR34 knockdown on the proliferation, migration, and apoptosis of HGC-27 gastric cancer cells and the underlying mechanisms.

METHODSThe expression of GPR34 in gastric cancer cell line HGC-27 was detected by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. HGC-27 cells were employed to construct the stable GPR34 knockdown cell model in this study. Real-time RT-PCR and Western blotting were applied to validate the effect of short hairpin RNA (ShRNA) on the expression of GPR34 in HGC-27 gastric cells. The proliferation, migration of these cells were examined by Cell Counting Kit-8 and transwell. We also measured expression profile of PI3K/PDK1/AKT and ERK using Western blotting.

RESULTSThe ShRNA directed against GPR34 effectively inhibited both endogenous mRNA and protein expression levels of GPR34, and significantly down-regulated the expression of PIK3CB (P < 0.01), PIK3CD (P < 0.01), PDK1 (P < 0.01), phosphorylation of PDK1 (P < 0.01), Akt (P < 0.01), and ERK (P < 0.01). Furthermore, GPR34 knockdown resulted in an obvious reduction in HGC-27 cancer cell proliferation and migration activity (P < 0.01).

CONCLUSIONSGPR34 knockdown impairs the proliferation and migration of HGC-27 gastric cancer cells in vitro and provides a potential implication for therapy of gastric cancer.

Apoptosis ; genetics ; physiology ; Blotting, Western ; Cell Line, Tumor ; Cell Proliferation ; genetics ; physiology ; Humans ; RNA, Small Interfering ; genetics ; Real-Time Polymerase Chain Reaction ; Receptors, Lysophospholipid ; genetics ; metabolism ; Stomach Neoplasms ; genetics ; metabolism

OBJECTIVETo identify a naturally presented HLA-A2-restricted epitope of MAGE-A3 antigen, FLWGPRALV (MAGE-A3(271 - 279)), on the surface of a human hepatocellular carcinoma (HCC) cell line HLE.

METHODSSynthetic peptide FLWGPRALV, served as positive control target, was analyzed by HPLC and HPLC-ESI-TOF-MSMS, in order to determine its HPLC elution time, mass-spectrometric characteristics and the lowest detection limitation by the two approaches. 3 x 10(9) HLE cells were collected, peptides naturally presented by major histocompatibility complex (MHC) molecules on the cell surface were isolated by mild acid elution, and concentrated by lyophilization, then the mixtures of peptides were fractioned by HPLC. The ingredient ranged from 2 min before the elution time determined by the synthetic peptide to 2 min after that was collected, concentrated by lyophilization, and analyzed by HPLC-ESI-TOF-MSMS, to identify the existence of the MAGE-A3(271 - 279) peptide.

RESULTSThe HPLC-ESI-TOF-MSMS detection provided an evidence for the existence of a doubly charged ion of (m/z)(2) 529.9, which was further analyzed by collision induced dissociation. The doubly charged ion was ultimately identified as the MAGE-A3(271 - 279) peptide, its amino sequence was FLWGPRALV and its molecular weight was 1058.4 Da.

CONCLUSIONSMAGE-A3(271 - 279) epitope could be naturally presented by HLA-A2 molecules to the surface of HCC cell line and MAGE-A3(271 - 279) peptide may have potential immunotherapeutic value in HCC patients.

Amino Acid Sequence ; Antigen Presentation ; Antigens, Neoplasm ; analysis ; isolation & purification ; Carcinoma, Hepatocellular ; immunology ; pathology ; Cell Line, Tumor ; Chromatography, High Pressure Liquid ; Epitopes, T-Lymphocyte ; analysis ; isolation & purification ; HLA-A2 Antigen ; immunology ; Humans ; Liver Neoplasms ; immunology ; pathology ; Mass Spectrometry ; Neoplasm Proteins ; analysis ; isolation & purification

OBJECTIVETo investigate reason and the management of portal vein thrombosis in patients with portal hypertension postoperatively.

METHODS329 patients with portal hypertension in liver cirrhosis who had splenectomy was reviewed from 1992 to 2001. In whom 43 (13.1%) patients with portal vein thrombosis postoperative were analyzed.

RESULTSIn these patients, except 1 died for portal vein phlebitis, all patients were recovered. There are 138 patients who underwent splenectomy or splenectomy and devascularization, 26 (18.8%) of them had thrombosis. 191 patients underwent splenectomy and portacaval or portasplenic shut, 17 (8.9%) of them had thrombosis. The data of these two groups have significant difference (chi(2) = 8.44, P < 0.01).

CONCLUSIONSThrombocytosis postsplenectomy as well as the changes of portal hemodynamics is the main reason of portal vein thrombosis. Portal vein thrombosis is also in association with the operative ways. Operation standardization, dynamic examining platelet count, routine color ultrasonography examining and early anticoagulation therapy are the effective methods in preventing and managing portal thrombosis postoperation for portal hypertension.

Adult ; Budd-Chiari Syndrome ; etiology ; prevention & control ; therapy ; Female ; Follow-Up Studies ; Humans ; Hypertension, Portal ; surgery ; Male ; Middle Aged ; Portal Vein ; pathology ; Postoperative Complications ; Vascular Surgical Procedures ; adverse effects ; methods

OBJECTIVETo observe the effects of galectin-3 on proliferation and apoptosis of hepatic stellate cells.

METHODSRT-PCR and Western blot were used to detect the expression of galectin-3 in hepatic stellate cells. Short hairpin DNA targeting galectin-3 of rat was was ligated into the recombinant vector pGCsilencer U6/Neo/GFP/shRNA plasmid. Then the plasmid was transfected into rat hepatic stellate cells. RT-PCR and Western blot were used to detect the interfering efficiency. Cell proliferation level was observed by CCK8 method at 24, 48 and 72 hours after transfection. Cell apoptosis was measured by Annexin V/PI-labeled flow cytometric analysis.

RESULTSExpression of galectin-3 in HSC was verified by both RT-PCR and Western blot. The recombinant vector was successfully constructed and verified, and was transfected into rat hepatic stellate cells. Western Blot and RT-PCR results demonstrated that the expression level of Galectin-3 was significantly down-regulated in galectin-3 shRNA transfected cells compared to control vector transferred cells. CCK8 assay indicated that proliferation of Galectin-3 knockdown cells was lower than that of control cells 48 and 72 hours post-transfection. Apoptotic cells in shRNA-interfering group were higher than those in control group both in early stage and advanced stage.

CONCLUSIONHepatic stellate cells can express galectin-3. Inhibition of galectin-3 using RNAi technique can suppress proliferation and induce apoptosis in HSC.

Animals ; Apoptosis ; Cell Line ; Cell Proliferation ; Down-Regulation ; Flow Cytometry ; Galectin 3 ; genetics ; metabolism ; Genetic Vectors ; Hepatic Stellate Cells ; cytology ; metabolism ; Liver Cirrhosis ; pathology ; Plasmids ; genetics ; RNA Interference ; RNA, Messenger ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Rats ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

OBJECTIVEKupffer cells (KCs) are resident macrophages in the liver. Because the densities and sizes of KCs show a significant overlap with other sinusoidal cells, it is difficult to separate and purify them. We aim to find an improved procedure that could optimize the method for isolation of highly purified rat Kupffer cells.

METHODSIn ex vivo rat liver perfusion with pronase E and collagenase IV, density gradient centrifugation by Histodenz and selective attachment of Kupffer cells were used to isolate them. Cell proliferation and morphological characterization were studied under light phase-contrast microscopes; KCs were also studied with transmission electron microscopy and scanning electron microscopy using standard techniques. Immunocytochemistry was used to detect the expression of ED2 CD163 and lysosome associated membrane protein 2 (LAMP2). Phagocytosis of latex-beads by KCs was also studied.

RESULTSThe yield rate of KCs was 5 x 10(7) and the KCs viability exceeded 98%. The purity of KCs identified by ED2 was higher than 98%, and over 99% of the collected KCs were LAMP2 positive.

CONCLUSIONIn the future this simple, stable and effective method of collecting highly purified Kupffer cells is expected to help in further studies.

Animals ; Cell Culture Techniques ; Cell Separation ; methods ; Cells, Cultured ; Kupffer Cells ; cytology ; Male ; Rats ; Rats, Inbred Lew