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1.
Article in Chinese | WPRIM | ID: wpr-700763

ABSTRACT

Hepatocellular carcinoma (HCC) is one of the most common malignancies in the world. For all the advances in surgical techniques,including transplantation, no substantial improvement has been made in the curative effect of HCC. The 1-year survival rate of the HCC patients with recurrence after liver transplantation is only 18%. In order to optimize the screening criteria for transplant recipients,perfect the distribution system of the donor,reduce the recurrence rate and improve the survival of the patients, it is necessary to gain a deeper insight into the tumor biological characteristics of the patients undergoing liver transplantation,precisely predict their prognostic indexes,and ascertain the relationship of post-transplant recurrence and metastasis with the patient's immune status, so as to carry out effective intervention for high-risk patients and individualized and integrated treatment for those with recur-rence or metastasis. Evidence shows that circulating tumor cells (CTC) and circulating tumor microemboli (CTM) are closely related with and contribute even more than other factors,such as the tumor size,tumor number,and vascular invasion,to the postoperative re-currence and metastasis of HCC. Therefore,a dynamic evaluation of CTC and CTM before liver transplantation can provide essential in-dicators for screening the recipient and predicting postoperative recurrence and metastasis.

2.
Basic & Clinical Medicine ; (12): 260-264, 2018.
Article in Chinese | WPRIM | ID: wpr-693882

ABSTRACT

Endoplasmic reticulum stress (ERS) is an important cellular self-protection mechanism. It can activate the unfolded protein response (UPR) that promotes cell survival, but sustained ERS may lead to cell apoptosis. There are a lot of endoplasmic reticulum in the liver cells, and many liver-related diseases are associaled with endoplasmic reticulum stress, such as alcoholic liver disease, non-alcoholic liver disease, toxic liver injury, viral hepatitis, hepatic malignant tumors and hepatic ischemia- perfusion injury. This article will summarize the role of ERS in the pathogenesis of liver-related diseases and the significance of intervention in liver disease.

3.
Chinese Journal of Oncology ; (12): 444-447, 2010.
Article in Chinese | WPRIM | ID: wpr-260381

ABSTRACT

<p><b>OBJECTIVE</b>To study the clinicopathological and immunohistochemical features, histogenesis and biological behavior, clinical treatment and prognosis of solid pseudopapillary tumor of the pancreas (SPT).</p><p><b>METHODS</b>Routine HE and immunohistochemical (SP) stainings were used in the pathological examination of 18 cases of SPT. Their clinical data were retrospectively analyzed. All the 18 postoperative patients were followed-up for 3 months to 10 years with an average of 29.2 months.</p><p><b>RESULTS</b>There were 16 females and 2 males, age ranging from 9 to 65 years with mean age of 25.3 years. Abdominal pain and palpable mass were among the major complains. Tumors were encapsulated and mixed with solid and cystic tissues. Histological features were pseudopapillary structure with a fibrovascular core. Immunhistologically, most tumors were positive for alpha-AT, alpha-ACT and Vim, with a high percentage of 94.4%. The eighteen cases were followed-up from 3 to 120 months. Five cases received reoperation after recurrence, and 14 cases were alive. Maximum survival time was 121 months and the minimum survival time was 3 months, with a median survival time of 23.0 months. The 5-year survival rate was 72.2%. A Kaplan-Meier analysis revealed that patient's age, tumor size, pathologic features, metastasis were major prognostic factors for SPT.</p><p><b>CONCLUSION</b>SPT is a tumor of low-grade malignancy and may be derived from multipotent stem cells. SPT most frequently affects young female, and has distinct clinicopathologic manifestation with excellent prognosis after surgical treatment.</p>


Subject(s)
Adolescent , Adult , Aged , Carcinoma, Papillary , Diagnosis , Metabolism , Pathology , General Surgery , Child , Female , Follow-Up Studies , Humans , Male , Middle Aged , Neoplasm Recurrence, Local , General Surgery , Pancreatectomy , Methods , Pancreatic Neoplasms , Diagnosis , Metabolism , Pathology , General Surgery , Reoperation , Retrospective Studies , Survival Rate , Vimentin , Metabolism , Young Adult , alpha 1-Antichymotrypsin , Metabolism , alpha 1-Antitrypsin , Metabolism
4.
Chinese Journal of Surgery ; (12): 1616-1619, 2009.
Article in Chinese | WPRIM | ID: wpr-291046

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the perioperative clinical outcome and predictive factors for perioperative complication morbidity and mortality.</p><p><b>METHODS</b>From August 2003 to August 2008, the data of 338 cases of hepatectomy performed in the liver transplant center of the First Affiliated Hospital of Nanjing Medical University was collected in a prospective manner. The patients' perioperative clinical risk factors and results were analyzed.</p><p><b>RESULTS</b>In the 338 hepatectomy cases, 255 patients (75.4%) underwent precise anatomical hepatectomy. The overall perioperative complication morbidity was 18.1%, while the perioperative mortality was 0.6%. In a total of 211 (62.4%) cases, the operation was carried out without blood transfusion. Univariate analysis revealed that cirrhotic liver, thrombocytopenia, blood loss in operation > 1000 ml, blood transfusion in operation and several other factors were closely related with the incidence rate of complication. Multivariate logistic regression analysis indicated that thrombocytopenia and perioperative blood transfusion were important independently predictive factors for the occurrence of perioperative complications in hepatectomy.</p><p><b>CONCLUSIONS</b>Precise hepatectomy enables patients to obtain better clinical outcome with low complication morbidity and perioperative mortality. Reducing hemorrhage is an important factor that lead to good clinical results.</p>


Subject(s)
Blood Loss, Surgical , Hepatectomy , Methods , Mortality , Humans , Intraoperative Complications , Epidemiology , Logistic Models , Minimally Invasive Surgical Procedures , Multivariate Analysis , Retrospective Studies , Risk Factors , Thrombocytopenia
5.
Chinese Journal of Surgery ; (12): 1658-1662, 2009.
Article in Chinese | WPRIM | ID: wpr-291036

ABSTRACT

<p><b>OBJECTIVE</b>To study the effect of total body irradiation of the donor in a spontaneous tolerance rat liver transplantation model and the role of CD4(+)CD25(+) regulatory T cells on induction of immunotolerance in the recipient.</p><p><b>METHODS</b>Liver transplantation was performed using male Lewis rats as donors and male DA rats as recipients. These rats were randomly allocated into the following groups:Control group, Homogeneity Liver Transplantation group, Idio-immunotolerance group and Acute Rejection group. After transplantation, survival time rate of each group were observed. Serum ALT, TB level, Foxp3(+)CD4(+)CD25(+) regulatory T cells, expression of GITR on T cell subgroup, histopathology of the hepatic graft on day 14, spleen CTL lytic activity on day 14 were measured.</p><p><b>RESULTS</b>In the Idio-immunotolerance group, the recipients suffered from transient rejection after surgery but acquired immunotolerance and survived long. In the Acute Rejection group, the donors were preconditioned with total body irradiation before liver transplantation. All recipients died between day 17 to 21. Serum ALT and TB increased significantly and the ratio of Foxp3(+)CD4(+)CD25(+) regulatory T cells decreased significantly compared with the Idio-immunotolerance group, the Homogeneity Liver Transplantation group and the Control group. The expression of GITR on CD3(+)CD4(+)T cells in the peripheral blood decreased, the expression of GITR on CD3(+)CD8(+) T cells and CTL lytic activity of the recipients increased by preconditioning of the donors with total body irradiation.</p><p><b>CONCLUSIONS</b>Preconditioning of the donors with total body irradiation eliminated the passenger lymphocytes of the liver graft, decreased the expression of Foxp3(+)CD4(+)CD25(+) regulatory T cells in peripheral blood, and increased the expression of GITR on CD3(+)CD8(+) T cells, thus affected the course of tolerance and induced acute rejection after liver transplantation.</p>


Subject(s)
Animals , Liver Transplantation , Allergy and Immunology , Male , Rats , Rats, Inbred Lew , T-Lymphocytes, Regulatory , Allergy and Immunology , Tissue Donors , Transplantation Conditioning , Transplantation, Homologous , Allergy and Immunology , Whole-Body Irradiation
6.
Chinese Journal of Surgery ; (12): 437-440, 2009.
Article in Chinese | WPRIM | ID: wpr-280638

ABSTRACT

<p><b>OBJECTIVE</b>To probe into indication of living-related liver transplantation (LRLT) for Wilson's Disease.</p><p><b>METHODS</b>From January 2001 to February 2007, thirty-seven living-related liver transplants were performed. A retrospective analysis was carried on outcome of those patients. The indications for LRLT were acute hepatic failure in 3 patients and chronic advanced liver disease in 32 patients including 13 patients with Wilsonian neurological manifestations. Two patients presented with severe Wilsonian neurological manifestations even though their liver functions were stable. According to the scoring system for evaluation of the neurological impairment in Wilson disease based on neurological signs and functions (total score was 30), the pre-transplantation score of those patients with neurological manifestations was 15.9 +/- 4.3 (n = 15).</p><p><b>RESULTS</b>Thirty-seven patients were followed up for 20 - 93 months. The survival rates of post-transplant patients and grafts at 1, 3, and 5 year were 91.9%, 83.8%, 75.7%, and 86.5%, 78.4%, 75.7%, respectively. Postoperative surgical complications occurred in 2 donors with bile leakage required drainage, in 2 recipients with hepatic thrombosis underwent retransplantation of cadaveric liver and in 1 recipient with hepatic stenosis required balloon dilatation. Neurological function was improved in all recipients and the score of posttransplantation at 6, 12, 18, 24, and 30 month was 17.5 +/- 3.7 (n = 13); 21.0 +/- 4.3 (n = 12); 23.9 +/- 3.9 (n = 10); 26.6 +/- 2.2 (n = 10) and 28.1 +/- 1.9 (n = 7) respectively.</p><p><b>CONCLUSIONS</b>Patients with acute hepatic failure or patients with severe liver disease unresponsive to chelation treatment should be treated with LRLT. Early transplantation in patients with an unsatisfactory response medical treatment may prevent irreversible neurological deterioration even though their liver function is stable.</p>


Subject(s)
Adolescent , Adult , Child , Female , Follow-Up Studies , Hepatolenticular Degeneration , General Surgery , Humans , Liver Failure , General Surgery , Liver Transplantation , Living Donors , Male , Nervous System Diseases , Retrospective Studies , Transplantation, Homologous , Treatment Outcome , Young Adult
7.
Chinese Journal of Oncology ; (12): 410-413, 2009.
Article in Chinese | WPRIM | ID: wpr-293102

ABSTRACT

<p><b>OBJECTIVE</b>To study the efficacy of anti-telomerase siRNA in hepatocellular carcinoma both in vitro and in vivo.</p><p><b>METHODS</b>Lentvirus vectors contained anti-telomerase siRNA were conducted with a high performance homologous recombination system, and then were transduced into human hepatocellular carcinoma HepG2 cells. The telomerase activity was detected by RT-PCR, HepG2 cell proliferation was determined by MTT assay, and apoptosis was detected by TUNEL assay. The in vivo experiment was carried out by inoculation of HepG2 cells into nude mice and the tumor growth was measured and analyzed.</p><p><b>RESULTS</b>The growth of transfected HepG2 cells was significantly inhibited and the inhibition rate was 57.5% at the 8th day after transfection. The telomerase activity was significantly suppressed in vitro. The growth of transfected human hepatocellular HepG2 tumor in the nude mice was also significantly inhibited.</p><p><b>CONCLUSION</b>The results of this study demonstrate that the growth of hepatocellular carcinoma cells is effectively inhibited by transfection of anti-telomerase siRNA both in vitro and in vivo.</p>


Subject(s)
Animals , Apoptosis , Carcinoma, Hepatocellular , Therapeutics , Cell Proliferation , Female , Genetic Therapy , Methods , Genetic Vectors , Hep G2 Cells , Humans , Lentivirus , Genetics , Liver Neoplasms , Therapeutics , Mice , Mice, Inbred BALB C , Mice, Nude , Neoplasm Transplantation , RNA, Messenger , Metabolism , RNA, Small Interfering , Recombinant Proteins , Genetics , Metabolism , Telomerase , Genetics , Metabolism , Transfection , Tumor Burden
8.
Chinese Journal of Hepatology ; (12): 363-367, 2009.
Article in Chinese | WPRIM | ID: wpr-310085

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the antiangiogenic property of pigment epithelium-derived factor(PEDF) in heptocarcinoma cell lines and explore its possible application in the gene therapy of human hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>The gene encoding human PEDF was subcloned into lentiviral vector to generate the recombinant plasmid pLenti-PEDF. The plasmid pLenti-PEDF and two other packaging plasmids were cotransfected to 293T cells by calcium phosphate. Then HepG2 was infected with recombinant lentivirus and the expression efficiency of PEDF was analyzed by western blot. Proliferation and migration assay of human umbilical vein endothelial cells (HUVEC) was used to evaluate the biological activity of PEDF in vitro. Murine subcutaneous tumor model was established to investigate the therapeutic effects of Lenti-PEDF on HCC, and the expression of PEDF mRNA in tumor tissues was analyzed by RT-PCR.</p><p><b>RESULTS</b>Restriction enzyme digestion and DNA sequencing demonstrated that the recombinant plasmid pLenti-PEDF was constructed successfully. HepG2 secreted PEDF in the media effectively after infected with the recombinant lentivirus and this protein exhibited strong inhibitory effects on proliferation and migration of human umbilical vein endothelial cells (P less than 0.01). Intratumoral injection of Lenti-PEDF caused significant inhibition of tumor growth (P less than 0.01), and high level expression of PEDF mRNA was detected in tumor tissues by RT-PCR.</p><p><b>CONCLUSIONS</b>Our data suggest that PEDF may exert an inhibitory effect on tumor angiogenesis and PEDF gene therapy may provide a new approach for the treatment of HCC.</p>


Subject(s)
Animals , Cell Proliferation , Endothelial Cells , Metabolism , Eye Proteins , Genetics , Metabolism , Genetic Therapy , Genetic Vectors , Hep G2 Cells , Humans , Lentivirus , Genetics , Liver Neoplasms , Genetics , Therapeutics , Male , Mice , Mice, Inbred BALB C , Mice, Nude , Neoplasm Transplantation , Neovascularization, Pathologic , Therapeutics , Nerve Growth Factors , Genetics , Metabolism , RNA, Messenger , Genetics , Metabolism , Serpins , Genetics , Metabolism , Transfection , Umbilical Veins , Cell Biology
9.
Chinese Journal of Oncology ; (12): 654-658, 2008.
Article in Chinese | WPRIM | ID: wpr-255609

ABSTRACT

<p><b>OBJECTIVE</b>To study the mechanism of enhancement of the CTL activity in mice co-expressing of CD80, CD86 and CD137L genes.</p><p><b>METHODS</b>The mice were randomly divided into five groups, named A, B, C, D and E. The group A and B were control groups (CG). H22-BAL B/c HCC mouse model was established by subcutaneous injection with hepatocellular carcinoma cells of cell line H22-Wt (group A), H22-neo (group B), H22-CD80/CD86(+) (group C), H22-CD137L(+) (group D) and H22-CD80/CD86/CD137L(+) (group E), respectively. On the 14th, 35th, 56th and 84th day after the first inoculation of tumor cells, TUNEL staining and DNA ladder examination were used to detect apoptosis of splenic T lymphocytes in all groups at each post-inoculation time point. Electrophoretic mobility-shift assay (EMSA) method was used to detect the activity of nuclear factor kappaB (NF-kappaB) in splenic T lymphocytes in each group at each time point post-inoculation.</p><p><b>RESULTS</b>Apoptosis was found in a great number of T lymphocytes in CG on the 14th day, much more than that in group C and E. The number of apoptotic T cells in group C had a significant difference compared with that in the group E from 14th to 84th day (P = 0.003). DNA ladder analysis showed typical positive results in group C and E. The significant apoptosis fragments were found in group C on 21st, 35th and 84th days. NF-kappaB activity of T cells in groups C and E was remarkably higher than that of groups CG and D, with higher in group D than that of CG (P = 0.002), and with no significant difference between group C and E on 14th day. The activity in group E was stable and remarkably higher than that of group C on 56th and 84th days after the first inoculation.</p><p><b>CONCLUSION</b>H22-CD80/CD86/CD137L(+) induces higher NF-kappaB activity of the host T cells by synergistic action of CD28 and CD137, which may be one of the mechanisms of enhancement of the host CTL activity induced by co-expression of CD80, CD86 and CD137L genes.</p>


Subject(s)
4-1BB Ligand , Metabolism , Animals , Apoptosis , B7-1 Antigen , Metabolism , B7-2 Antigen , Metabolism , CD28 Antigens , Cell Line, Tumor , Female , Gene Expression Regulation, Neoplastic , Liver Neoplasms, Experimental , Allergy and Immunology , Metabolism , Pathology , Lymphocyte Activation , Mice , Mice, Inbred BALB C , NF-kappa B , Metabolism , Random Allocation , Spleen , Pathology , T-Lymphocytes , Metabolism , Pathology , T-Lymphocytes, Cytotoxic , Allergy and Immunology
10.
Chinese Journal of Surgery ; (12): 1019-1022, 2007.
Article in Chinese | WPRIM | ID: wpr-340869

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the outcome of emergency adult right lobe living donor liver transplantation for fulminant hepatitis.</p><p><b>METHODS</b>Nine cases of adult right lobe living donor liver transplantation were performed from September 2002 to August 2005, the clinical and follow-up data was analyzed.</p><p><b>RESULTS</b>According to Child Pugh Turcotte (CPT) classification, 9 patients were classified as grade C before transplant. The Model for End-Stage Liver Disease (MELD) scores of these patients were 26.7 +/- 8.8. The principal pre-transplant complications included hepatic encephalopathy (5 cases), electrolyte disturbance (3 cases), renal failure (2 cases), gastrointestinal bleeding (1 case). The operations in donors and recipients were all successful. The post-transplant complications induced pulmonary infection in 2 patients, acute renal failure in 3 and transplantation related encephalopathy in 1. There were no primary graft non-function and no blood vessel and bile tract complications occurred. One-year survival rate was 55.6%. No serious complication or death found in donors.</p><p><b>CONCLUSIONS</b>Emergency adult to adult living donor liver transplantation is an effective treatment for fulminant hepatitis but the safety of the donors should be assessed strictly preoperation.</p>


Subject(s)
Adult , Critical Illness , Emergency Medical Services , Female , Follow-Up Studies , Hepatitis , Pathology , General Surgery , Humans , Liver Transplantation , Methods , Living Donors , Male , Middle Aged , Retrospective Studies , Treatment Outcome
11.
Chinese Journal of Surgery ; (12): 323-325, 2007.
Article in Chinese | WPRIM | ID: wpr-342173

ABSTRACT

<p><b>OBJECTIVE</b>To study the substitute portal vein by irradiated allograft saphenous vein during liver transplantation and investigate the changes in morphology and immunology.</p><p><b>METHODS</b>All the recipients were divided into 3 groups randomly:irradiated allograft group (n = 11) (group A), fresh allograft group (n = 9) (group B) and fresh self-graft group (n = 14) (group C). The number of non-jam graft vessels in each group was explored at 1st week, 2nd week, 1st month, 2nd month and 3rd month post-operation. Also, the infiltration of CD(4)(+), CD(8)(+) T cells and histological changes in grafted vessels were detected.</p><p><b>RESULTS</b>No obvious histological changes were observed in group A, as well as under naked eyes. There were 9, 3 and 12 non-jam vessels in group A, B and C and there were significant differences between group A and B (P < 0.05). The endothelial cells of graft vessels were observed both in group A and C two weeks post-operation and covered the graft vessels two months later. There were infiltration of lymphocytes and inflammatory cells at early stage, obvious damage and no endothelial cells growth in graft vessels in group B. Compared with group B, the percentage of CD(4)(+), CD(8)(+) T cells in group A was lower significantly, but higher slightly than that in group C.</p><p><b>CONCLUSIONS</b>Irradiated allograft saphenous veins have the quality of ideal vascular transplantation prosthesis and weak antigenicity at the same time. The changes of CD(4)(+), CD(8)(+) T cells after allograft vessels can be detected as immunology index for acute immunological rejection.</p>


Subject(s)
Adult , CD4-Positive T-Lymphocytes , Allergy and Immunology , CD8-Positive T-Lymphocytes , Allergy and Immunology , Graft Rejection , Diagnosis , Humans , Liver Transplantation , Male , Neutrophil Infiltration , Saphenous Vein , Allergy and Immunology , Radiation Effects , Transplantation , Transplantation, Homologous
12.
Article in Chinese | WPRIM | ID: wpr-336481

ABSTRACT

<p><b>OBJECTIVE</b>To identify genetic abnormalities in primary gastric carcinoma.</p><p><b>METHODS</b>Comparative genomic hybridization (CGH) was used in screening DNA copy number changes along all chromosomes in 23 cases of primary gastric cancer.</p><p><b>RESULTS</b>Twenty-one out of 23 cases showed chromosomal losses and gains for at least one of the chromosomal arms in primary gastric cancer. The mean number of chromosomal alterations was 7.52. Chromosomal gains predominated over chromosomal losses in a ratio of 5.38:2.14. The most often involved chromosomal gains were observed in 8q (9/21, 42.9%), 20q (9/21, 42.9%), 17q (8/21, 38.1%), 3q (7/21, 33.3%), 7q (7/21, 33.3%), 11q (6/21, 28.6%), 13q (6/21, 28.6%), 1q (5/21, 23.8%) and 20p (5/21, 23.8%). The chromosomal arms with frequent losses were 17p (7/21, 33.3%), 18q (6/21, 28.6%), 5q (5/21, 23.8%), 8p (5/21, 23.8%), and 9p (5/21, 23.8%).</p><p><b>CONCLUSIONS</b>The phenomenon of gain and loss of chromosomal regions is observed in primary gastric cancer, which may induce the amplification of oncogenes and the loss of tumor suppressor genes to regulate the development and progression of gastric cancer.</p>


Subject(s)
Adult , Aged , Aged, 80 and over , Chromosome Aberrations , Chromosome Deletion , Comparative Genomic Hybridization , DNA , Female , Gene Expression Profiling , Genes, Tumor Suppressor , Humans , In Situ Hybridization, Fluorescence , Male , Middle Aged , Neoplasm Staging , Stomach Neoplasms , Genetics , Pathology
13.
Chinese Journal of Surgery ; (12): 1448-1452, 2006.
Article in Chinese | WPRIM | ID: wpr-288573

ABSTRACT

<p><b>OBJECTIVE</b>To investigate and evaluate different surgical methods applied in living-donor liver transplantation (LDLT).</p><p><b>METHODS</b>Fifty patients with end-stage liver disease received LDLT in our department between January 1995 and March 2006. The data were analyzed on a retrospective basis. The choice of different surgical methods, strategies applied to ensure the safety of donors and indications of LDLT in the series were reviewed.</p><p><b>RESULTS</b>All donors recovered uneventfully. Among the 50 patients, 47 recipients presented with end-stage cirrhosis, 3 patients suffered from malignant tumor. To date, 6 recipients died after LDLT, among them, 3 recipients died of the operation and the other 3 recipients died of long-term complications. Resected donor livers included 9 cases of segments V, VI, VII and VIII (not including the middle hepatic veins) and 1 case of segments V, VI, VII and VIII (including the middle hepatic veins), 36 cases of segments II, III and IV (including the middle hepatic veins) and 4 cases of segments II, III, and part of IV (not including middle hepatic veins).</p><p><b>CONCLUSIONS</b>LDLT helps tackle the problem of donor shortage in the world. The process is complicated, and it is very important to choose appropriate surgical methods for the improvement of surgical achievement and donor safety.</p>


Subject(s)
Adolescent , Adult , Child , Child, Preschool , Female , Follow-Up Studies , Humans , Infant , Liver Cirrhosis , General Surgery , Liver Neoplasms , General Surgery , Liver Transplantation , Methods , Living Donors , Male , Middle Aged , Retrospective Studies
14.
Chinese Journal of Surgery ; (12): 1463-1466, 2006.
Article in Chinese | WPRIM | ID: wpr-288569

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the role of intrahepatic CD4(+)CD25(+) T regulatory cells and Foxp3 gene in the natural tolerance in rat liver transplantation.</p><p><b>METHODS</b>The orthotopic liver transplantation models of inbred rats (LEW and DA rats) were established with double-sleeve technique and the models were divided into two groups: tolerance group (TOL group, LEW-to-DA) and rejection group (REJ group, DA-to-LEW). The intrahepatic lymphocytes from each group were isolated by using density gradient centrifugation. CD4(+)CD25(+) T cells were isolated by magic cell sorting system (MACS) and identified by flow cytometry (FCM). CD4(+)CD25(+) Tr cells suppression on the proliferation of CD4(+)CD25(-) T effector cells were analyzed by cell proliferation assay in vitro. Western blot was used to detect Scurfin protein expression of CD4(+)CD25(+) Tr cells.</p><p><b>RESULTS</b>CD4(+)CD25(+) Tr cells developed significantly greater in the TOL group than in the REJ group. In vitro, the spleen cells from LEW rats can irritate the proliferation of CD4(+)CD25(+) T cells more obviously than the syngeneic spleen cells. CD4(+)CD25(+) T cells could suppress the proliferation of CD4(+)CD25(-) T cells, but the inhibition was reversed by exogenous IL-2 (200 U/ml).</p><p><b>CONCLUSIONS</b>The immune suppression function of CD4(+)CD25(+) Tr cell, mediated by Foxp3 gene, is one of the mechanisms in liver transplantation tolerance.</p>


Subject(s)
Animals , Blotting, Western , CD4-Positive T-Lymphocytes , Cell Biology , Allergy and Immunology , Metabolism , Cells, Cultured , Flow Cytometry , Forkhead Transcription Factors , Metabolism , Graft Rejection , Allergy and Immunology , Interleukin-2 Receptor alpha Subunit , Allergy and Immunology , Liver Transplantation , Allergy and Immunology , Male , Models, Animal , Rats , Rats, Inbred Lew , Rats, Inbred Strains , Transplantation Tolerance , Allergy and Immunology
15.
Chinese Journal of Hepatology ; (12): 824-827, 2006.
Article in Chinese | WPRIM | ID: wpr-260582

ABSTRACT

<p><b>OBJECTIVES</b>To analyze the differential expression genes (DEGs) among hepatocellular carcinoma (HCC), para-cancerous tissue (PCT) and normal liver tissue (NLT) and explore the target genes related to the development and progression of HCC.</p><p><b>METHODS</b>The total RNAs of matched HCC, PCT and NLT of HCC patients were isolated using one step Trizol method. Matched RNAs were qualified using 10 g/L agarose gel electrophoresis and lab-on-chip. cRNAs were synthesized, fluorescence labeled and purified after total RNAs were purified. The RNAs of HCC and NLT, HCC and PCT were hybridized with Agilent oligo microarray (21,074 probes). The fluorescence intensity features were detected by Agilent scanner and quantified by feature extraction software. The selected candidate genes were confirmed by SYBR Green I stained real time RT-PCR.</p><p><b>RESULTS</b>(1) The total RNA, reverse transcription product and fluorescence labeled cRNA were all of high quality; (2) There were 420 up-regulated genes and 552 down-regulated genes among 2-fold DEGs, including DKK1 (dickkopf homolog 1) which was 5-fold up-regulated; (3) The results of real time RT-PCR, using beta-actin as an internal control, showed that the 2-Delta Ct values of DKK1 in HCC, PCT and NLT were 0.089 504, 0.007,65 and 0.000,631 respectively.</p><p><b>CONCLUSION</b>(1) The high throughput and effective Agilent oligo microarray can screen novel therapy targeted genes by analyzing the DEGs in development and progression of HCC; (2) The development and progression of HCC is a complicated process involving multigenes and multiprocedures; (3) DKK1, as a novel gene, is involved in the development and progression of HCC and may be a new therapy target.</p>


Subject(s)
Carcinoma, Hepatocellular , Genetics , Pathology , Gene Expression , Gene Expression Regulation, Neoplastic , Humans , Liver Neoplasms , Genetics , Pathology , Oligonucleotide Array Sequence Analysis , RNA, Neoplasm , Genetics
16.
Chinese Journal of Hepatology ; (12): 243-246, 2006.
Article in Chinese | WPRIM | ID: wpr-245690

ABSTRACT

<p><b>OBJECTIVE</b>To summarize our clinical experience in adult-to-adult living donor liver transplantation (ALDLT).</p><p><b>METHODS</b>Clinical data of 12 patients with ALDLT performed in our center from September 2000 to June 2005 were analyzed, retrospectively.</p><p><b>RESULTS</b>Left lobe (segments II, III, IV, including the middle hepatic veins) transplantation was performed in 3 patients and right lobe (segments V, VI, VII, VIII, with or without the middle hepatic veins) transplantation was performed in 9 patients. Donors: There were no operative deaths. The median operative time was 6.20+/-1.40 hours and their blood loss ranged from 300 ml to 1200 ml. Postoperative complications included biliary fistula (1 donor) and wound fat liquefaction (1 donor). During a 6-12 months follow-up, no long-term complications were found. Recipients: The operating time ranged from 5 to 11 hours and their blood loss ranged from 800 to 7000 ml. Modified outflow reconstruction, microvascular reconstruction of the hepatic artery and duct-to-duct biliary reconstruction were done during the recipient operations. The median cold ischemia time was 1.90+/-0.50 hours. The median anhepatic phase of recipients was 1.63+/-0.43 hours. Graft/recipient weight ratio (GRWR) was (1.20+/-0.26)%. One recipient presented a postoperative complication of biliary fistula and another recipient died 1 month after the operation from serious infection. The other 11 recipients had long-term survivals.</p><p><b>CONCLUSION</b>ALDLT is an effective treatment for decompensated end-stage liver disease patients and is relatively safe for the donors.</p>


Subject(s)
Adult , Female , Hepatolenticular Degeneration , General Surgery , Humans , Liver Cirrhosis , General Surgery , Liver Transplantation , Living Donors , Male
17.
Chinese Medical Journal ; (24): 1003-1009, 2006.
Article in English | WPRIM | ID: wpr-265263

ABSTRACT

<p><b>BACKGROUND</b>Because of the lack of brain death laws in China, the proportion of cadaveric organ donation is low. Many patients with end-stage liver disease die waiting for a suitable donor. Living donor liver transplantation (LDLT) would reduce the current discrepancy between the number of patients on the transplant waiting list and the number of available organ donors. We describe the early experience of LDLT in the mainland of China based on data from five liver transplant centers.</p><p><b>METHODS</b>Between January 2001 and October 2003, 45 patients with end-stage liver disease received LDLT at five centers in China. The indication and timing, surgical techniques and complications, nonsurgical issues including rejection, infection, and advantages of LDLT in the series were reviewed. Actuarial patient and graft survival rates were calculated by using the Kaplan-Meier product-limit estimate. Statistical analysis was completed by using SPSS 10.0.</p><p><b>RESULTS</b>All LDLT recipients were cirrhotic patients, except for one man with fulminant hepatic failure. Among the 45 cases of LDLT, 35 (77.8%) were performed in one center (the First Affiliated Hospital of Nanjing Medical University). The overall 1 and 3 year survival rate of the recipients was 93.1% and 92.0%, respectively. Of the 45 LDLT donors, there were 3 cases of biliary leakage, 2 subphrenic collections, 1 fat liquefaction around the incision and 1 biliary peritonitis after T tube removal. All donors recovered completely.</p><p><b>CONCLUSIONS</b>LDLT provides an excellent approach to addressing the problem of donor shortage in China even though the operation is complicated, uncompromising and difficult with respect to the safety of the donors and receptors. Despite early technical hurdles having been overcome, perfection of technique is still necessarily. At present, LDLT is a good choice for the patients with irreversible liver disease.</p>


Subject(s)
Adolescent , Adult , Child , Child, Preschool , Humans , Liver Transplantation , Living Donors
18.
Chinese Journal of Oncology ; (12): 490-493, 2006.
Article in Chinese | WPRIM | ID: wpr-236951

ABSTRACT

<p><b>OBJECTIVE</b>To understand the influence of co-expression of CD80, CD86 and CD137L genes on tumor immunogenicity in hepatocellular carcinoma H22-BAL B/c mouse models.</p><p><b>METHODS</b>The mice were randomly divided into five groups, named A, B, C, D and E, and control groups A and B, 20 mice in each group. Hepatocellular carcinoma H22-BAL B/c mouse model was established by subcutaneous injection of cells H22-Wt, H22-neo, H22-CD80/CD86+, H22-CD137L+ and H22-CD80/CD86/CD137L+, respectively. The rate and incubation period of tumor development, survival rate, and the tumor growth in vivo were observed and recorded. The effects of gene transduction on immunogenicity of the tumor and antitumor immunity of the animals were assessed by re-innoculation of wild type H22 cells.</p><p><b>RESULTS</b>The rate of tumor development in group E was only 50%, much lower than that in other four groups (P < 0. 01). The tumor growth in group C was reduced with complete tumor regression in two hosts (20%, 2/10). In group E, there was more pronounced reduction of tumor size. The maximal tumor sizes were remarkably smaller than those of group C, and there was complete tumor regression in three mice (60%, 3/5). No tumor regression was found in the other three groups. Survival rates of group C and E were significantly higher than that of animals in groups A, B and D (P < 0. 01), but no significant difference was seen between group C and E. The results of re-inoculation test showed that tumor formation rate was 40% (4/10) in group C, 100% (8/8) in group D, and 0 (0/5) in group E. There were significant differences between groups C and E and control group, between group E and C, but not between C and D. After the third time of re-inoculation with H22-Wt cells at the 56th day, tumor occurred in 6/6 mice (100%) of group C, but 0 (0/5) in group E. The difference was very significant. Five animals without tumor formation after the first inoculation in group E, were re-inoculated with H22-Wt cells on the 21st day and the third re-inoculation on the 56th day, no tumor was found (0/5).</p><p><b>CONCLUSION</b>Both co-expression and solo-expression of CD80+ CD86 and CD137L genes reduce the tumorigenicity of wild type H22 cells, but co-expression of CD80, CD86 and CD137L genes can more significantly improve the immunogenicity of H22-CD80/CD86/CD137L+ cells.</p>


Subject(s)
4-1BB Ligand , Genetics , Allergy and Immunology , Metabolism , Animals , B7-1 Antigen , Genetics , Allergy and Immunology , Metabolism , B7-2 Antigen , Genetics , Allergy and Immunology , Metabolism , Cell Line, Tumor , Female , Gene Expression Regulation, Neoplastic , Liver Neoplasms, Experimental , Genetics , Allergy and Immunology , Pathology , Mice , Mice, Inbred BALB C , Neoplasm Transplantation , Random Allocation , Survival Analysis
19.
Chinese Journal of Hepatology ; (12): 183-186, 2005.
Article in Chinese | WPRIM | ID: wpr-349174

ABSTRACT

<p><b>OBJECTIVES</b>To construct a recombinant adeno-associated virus vector pSNAV expressing CTLA-4Ig and to demonstrate its expression in transplanted liver allografts and to see if a long term inhibitive effect of CTLA-4Ig could be obtained though its use.</p><p><b>METHODS</b>After AAVCTLA-4Ig and PUC18 were cut with BamHI, CTLA-4Ig cDNA was inserted into the plasmid PUC18 by T4DNA ligase and PUC18-CTLA-4Ig was constructed. The obtained PUC18-CTLA-4Ig and pSNAV cut with Kpn I and EcoR I, CTLA-4Ig cDNA was inserted into plasmid pSNAV to construct the recombinant vector pSNAV-CTLA-4Ig, which was transfected into BHK-21 packaging cells by lipofectine-mediated transfection. Then the BHK-21 cell line was infected with HSV1-rc to produce a large amount of pSNAV- CTLA-4Ig. The specificity of the expressed product was identified by digestion with BamHI, PCR and sequence determination. The titer of the virus was detected. The product was infused into rats liver allografts via portal vein and its expression in the transplanted livers was detected immunohistochemically.</p><p><b>RESULTS</b>Recombinant adeno-associated virus vector pSNAV-CTLA-4Ig was generated and purified into 8.5 x 10(11)/ml. Agarose gel analysis of PCR products verified the presence of CTLA-4Ig. Digestion with BamHI and sequence determination confirmed that pSNAV-CTLA-4Ig was constructed. Expression of CTLA-4Ig in the transplanted livers was detected successfully.</p><p><b>CONCLUSION</b>Prepared pSNAV-CTLA-4Ig was constructed correctly and can express CTLA-4Ig effectively. Besides this, it can express CTLA-4Ig in rat liver allografts. It may be used in the study of transplant tolerance.</p>


Subject(s)
Abatacept , Animals , Dependovirus , Genetics , Metabolism , Genetic Vectors , Immunoconjugates , Genetics , Metabolism , Liver Transplantation , Random Allocation , Rats , Rats, Sprague-Dawley , Rats, Wistar , Transplantation, Homologous
20.
Chinese Journal of Oncology ; (12): 328-331, 2003.
Article in Chinese | WPRIM | ID: wpr-347433

ABSTRACT

<p><b>OBJECTIVE</b>To study the construction of the PCI-neo mammalian expression vector system containing murine 4-1BBL gene and its stable and effective expression in rat hepatocellular carcinoma cell line CBRH7919.</p><p><b>METHODS</b>The murine full-length 4-1BBL cDNA was obtained and subcloned into the PCI-neo mammalian expression vector. The recombinant named as PCI-neo-4-1BBL was identificated by restriction enzyme digestion and sequencing. Subsequently, PCI-neo-4-1BBL was transfected into CBRH7919 with lipofectamine reagent, then G418-resistance clone was acquired and named as PCI-neo-4-1BBL-CBRH7919+. The stable expression of 4-1BBL mRNA in PCI-neo-4-1BBL-CBRH7919+ was detected by RT-PCR.</p><p><b>RESULTS</b>The fragment of 980 bp (4-1BBL) and 5.4 kb (PCI-neo) was shown after PCI-neo-4-1BBL had been digested by EcoR I and Not I and agarose gel electrophoresis. The DNA sequencing of 4-1BBL, proved to be identical to the data of 4-1BBL in Genebank, showed stable and effective expression in PCI-neo-4-1BBL-CBRH7919+.</p><p><b>CONCLUSION</b>The PCI-neo mammalian expression vector system containing murine 4-1BBL has been constructed successfully, which shows stable and effective expression in CBRH7919.</p>


Subject(s)
4-1BB Ligand , Animals , Cell Line, Tumor , Cloning, Molecular , DNA, Complementary , Genetics , Genetic Vectors , Liver Neoplasms, Experimental , Metabolism , Pathology , RNA, Messenger , Genetics , Rats , Rats, Wistar , Recombination, Genetic , Transfection , Tumor Necrosis Factor-alpha , Genetics
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