AIM:To detect tissue factor(TF)level both in plasma and in tissue of hepatocellular carcinoma(HCC)patients and to elucidate their association with clinical features.METHODS:Plasma TF levels of 50 cases of HCC patients and 30 cases of control were detected by ELISA.27 HCC tissue samples with their adjacent tissue samples and 27 normal liver tissues were detected by RT-PCR.RESULTS:① Plasma TF levels were increased significantly in HCC group when compared with control(P

AIM: To study the expression of laminin and survivin in primary gallbladder carcinoma(PGC),and their correlation with clinicopathologic characteristics of PGC.METHODS: Forty-nine specimens with PGC,21 with cholecystoadenoma and 13 with chronic cholecystitis were included in this study.Laminin and survivin expression in gallbladder tissues were detected by immunohistochemistry.The relationship between laminin or survivin expression and clinico-pathologic data were also analyzed.RESULTS: An intact basement membrane(BM) with well-delineated,smooth and continuous line-like laminin staining was identified in the tissues from chronic cholecystitis,adenoma of gallbladder and gallbladder carcinoma in situ.Submucosal BM,which was interrupted in PGC specimens in NevinⅡstage,was almost lost in tumor tissues in Nevin Ⅲ,Ⅳ and Ⅴ stage.Meanwhile,a distinguishing feature of the tumor tissue was broken or discontinuous line-like laminin staining in the matrix to tumor cells.These lines were detected to be weak,or even entirely absent in some neoplastic tissue sections.Sometimes,a faint cytoplasmic staining of laminin was also found in tumor tissues as classified to Nevin Ⅲ,Ⅳ and Ⅴ stage.Expression of survivin in PGC tissues was significantly higher than that in adenoma of gallbladder and chronic cholecystitis.However,survivin expressions had no specificity and positive predictive value for cell differentiation and grade as well as clinic stage of PGC by using statistical analyses.Furthermore,no correlation was confirmed between the staining features of laminin and survivin expression.CONCLUSION: Laminin expression is strongly associated with the malignancy and invasiveness of PGC,and survivin might play an important synergistic role in the development of PGC.

AIM: To investigate whether viable apoptotic cells and phagocytosis of them affect the activation of T lymphocytes. METHODS: Ultraviolet irradiation was used to induce apoptotic cells in vitro and the model of phagocytosis of these cells was established. Cytokine TGF ?1 was detected by ELISA. The rate of apoptotic cells and phagocytosis of them were assessed by flow cytometry. Furthermore, flow cytometry was also employed to examine the expression of activation signs, such as CD69, CD25, CD71, of T lymphocytes under the intervention of apoptotic cells and macrophage which ingested apoptotic cells, to reflect whether the apoptotic cells and the phagocytosis of these cells could influence the activation of lymphocytes stimulated by Con A. RESULTS: Ingestion of apoptotic cells increased TGF ?1 secretion. Only the macrophages that had ingested apoptotic cells could suppress the activation of lymphocytes. The expression of the markers of lymphocytes activation such as CD69, CD25, CD71 had been restrained. These inhibition effects were abolished by monoclonal anti-TGF ?1 antibody. CONCLUSION: The macrophages that have ingested apoptotic cells inhibit expression of CD69, CD25 and CD71 of T lymphocytes stimulated by ConA. This effect is dependent on the increase in TGF ?1 secretion in local site. [

0.05). However, a significant higher percentage of CD3+CD25+ T cells was found in stable liver transplantation group as compared to healthy group ( P0.05). There was not found no regular change of serum cytokine s (IL-6, TN F-?) and adhesion molecules (ICAM-1, P-selectin) in 6 liver transplanted patien ts during post-operation from day 1 to day 30, indicating that was associated wi th the different status of patients before or after transplantation. CONCLUSIONS: Our data suggesting that increased levels of ICAM-1 and P-selectin, appears to participate in the processing of immunoregulation to transplanted livers, whereas elevated concentrations of IL-6 appear to be involv ed in the repair of the injury induced by TNF-? in allo-transplanted livers.

AIM: To investigate lymphotactin (Lptn) gene transcription during acute cardiac allograft rejection and the inhibitory effect of cyclosporine A (CsA). METHODS: Graft specimens were harvested at indicated time to determine morphological changes by pathological examination. The grade of acute cardiac allograft rejection was evaluated by using modified Banff scoring system. Reverse transcription-polymerase chain reaction (RT-PCR) was used to determine the Lptn mRNA expression in cardiac grafts. NFATc1 activity of splenocytes after transplantation was assessed by enzyme-linked immunoabsordent assay (ELISA). RESULTS: Prominent splenomegaly on day 3 posttransplantation was found in C57BL/6-Balb/c group. The extent of myocardial inflammatory infiltration was scored 2.667?0.577 at day 5 and 2.333?0.577 at day 7, respectively. Splenomegaly was ameliorated by CsA treatment, and the extent of myocardial infiltrate was scored 1.000?0.000 at day 5 and 1.333?0.577 at day 7, respectively. Lymphotactin mRNA was undetectable in cardiac isografts. Lymphotactin mRNA, which was inhibited partially by CsA, was upregulated strongly in acutely rejecting cardiac allografts at day 5 and day 7. Further studies demonstrated that NFATc1 activity in splenocytes, which markedly upregulated during acute rejection, was completely inhibited by CsA. CONCLUSION: Lptn appears to be a key chemokine of lymphocyte infiltration during acute allograft rejection. Inhibition of NFATc1 activity by CsA seems to decrease Lptn expression incompletely, suggesting that there was else mechanism to regulate Lptn expression other than NFAT pathway.

0.05).Tumor volumes was diminished in group B and C as compared with that in group A(P

AIM: To investigate the in vitro anti tumo r immune responses of dendritoma formed by mouse hepatocellular carcinoma cells and lymphotactin gene modified dendritic cells (DCs). METHODS: DCs prepared from mouse bone marrow were genetically mo dified by lymphotactin adenovirus, and fused with H22 cells by polyethylene glyc ol. RT-PCR and ELISA were employed to identify lymphotactin expression at mRNA a nd protein levels. The phenotypes and fusion efficiency were detected by FACS. T he stimulatory capacity of DC to T cells was detected by mixed leukocyte reactio n. The cytotoxicity activity against H22 cells was assayed by LDH method. RESULTS: Lymphotactin effectively expressed by DCLptn/H22 hybrid oma. DCLptn/H22 cells induced potent T cell proliferation effect and generated s trong CTL reaction against allogenic H22 cells. CONCLUSION: Lymphotactin genetic modification enhanced the in vitro immune activity of dendritoma.

AIM: To investigate the effects of alanyl-glutamine (Ala-Gln) on expression of iNOS and TNF-? in injured intestinal mucosa induced by oral tacrolimus(FK506). METHODS: Twenty-four BALB/c mice were randomized to receive orally 0.2 mL of normal saline solution ( groupⅠ), 0.2 mL of FK506 in a dose of 0.1 mg/kg (groupⅡ) or 1.0 mg/kg (groupⅢ), and orally high-dose FK506 (0.2 mL, 1.0 mg/kg) plus intraperitoneal injection of Ala-Gln (0.5 g/kg )(groupⅣ),respectively. Damages of intestinal mucosa were determined by pathological examination. Intestinal mucosal permeability was analysed by FITC-dextran fluorescence assay. Expression of iNOS and TNF-? in intestine was detected by RT-PCR and Western blotting.RESULTS: Severe damage on the villi and increased intestinal permeability were observed in high-dose FK506 treated mice according to scanning electron microscopy and FITC-dextran flux respectively. The erosion and increased intestinal permeability were significantly alleviated by Ala-Gln treatment. Transcription of iNOS mRNA and TNF-? mRNA, which was up-regulated in high-dose FK506 treated group, was also markedly down-regulated in mice combined with Ala-Gln-treatment. A significantly increased expression of iNOS and TNF-? protein was found in the high-dose FK506 treated mice, while small amounts of these proteins were identified in the Ala-Gln-treated group.CONCLUSION: FK506 could induce a significant impairment of intestinal mucosa morphologically, which might be associated with up-regulated expression of iNOS and TNF-? in small intestinal mucosa. Subsequently, the intestinal permeability is increased. Ala-Gln has a strong protective effect on FK506-induced intestinal barrier dysfunction, probably relates to the down-regulation of iNOS and TNF-? expression.

BACKGROUND: Extensive liver resection or liver transplantation operated on patients with combined hepatic cirrhosis and other complications correlates with high morbidity and mortality.Child-Turcotte-Pugh scoring system is now widely used in the assessment of liver function.This classification scheme includes three clinical indicators and two biochemical indices;however,it seems difficulty on directly evaluating functional status of hepatocytes.OBJECTIVE: To explore the practicability of bioluminescence adenosine triphosphate (ATP) determination assay to assess the functional reserve of residual hepatocytes,DESIGN,TIME AND SETTING: Case contrast study,which was carried out in the Second Affiliated Hospital,Sun Yat-sen University from January 2005 to March 2006.PARTICIPANTS: Thirty-two patients who underwent major extra-and intra hepatic surgery including liver transplantation were randomly divided into three groups based on hepatic cirrhosis grading standard,including normal group (n=7),macronodular cirrhosis group (n=9),and micronodular cirrhosis group (n=16).METHODS: Routine examination and biochemical indexes of liver were performed preoperatively,including glutamic oxalacetic transaminase (GOT) and total bilirubin (TBIL).Liver specimens were delivered by aseptic technique during operation and enzymatic digested.Cell suspension was cultured and centrifuged.Hepatocytes were counted and dispensed cell suspension to be used for ATP extraction and measurement.MAIN OUTCOME MEASURES: ATP content,preoperative biochemical parameters of liver function,and correlation between biochemical parameters and ATP content.RESULTS: The ATP content in the macronodular cirrhosis group was significantly higher than that in the micronodular cirrhosis group and normal group (P=0.000 1,0.004).While,the ATP content in the micronodular cirrhosis group was also significantly higher than that in the normal group (P=0.004).ATP content (mole/cell) wassignificantly positively correlated with serum glutamic oxalacetic transarninase (r=-0.609 3,P=0.000 2) and TBIL (r=0.614 5,P=0.000 2).CONCLUSION: ATP assay can directly evaluate functional reserve of liver parenchyma and reflect high operative risk status (HORS) and course of postoperative recovery in major hepatic resection.

AIM: To investigate the effects of alanyl - glutamine ( Ala -Gln) on expression of iNOS and TNF- α in injured intestinal mucosa induced by oral tacrolimus (FK506). METHODS: Twenty -four BALB/c mice were randomized to receive orally 0.2 mL of normal saline solution ( group Ⅰ ), 0.2 mL of FK506 in a dose of 0.1 mg/kg ( group Ⅱ ) or 1.0 mg/kg (group Ⅲ), and orally high -dose FK506 (0.2 mL, 1.0 mg/kg) plus intraperitoneal injection of Ala -Gln (0.5 g/kg )(group Ⅳ ),respectively. Damages of intestinal mucosa were determined by pathological examination.Intestinal mucosal permeability was analysed by FITC - dextran fluorescence assay. Expression of iNOS and TNF - α in intestine was detected by RT - PCR and Western blotting. RESULTS: Severe damage on the villi and increased intestinal permeability were observed in high - dose FK506 treated mice according to scanning electron microscopy and FITC - dextran flux respectively. The erosion and increased intestinal permeability were significantly alleviated by Ala - Gln treatment. Transcription of iNOS mRNA and TNF - α mRNA, which was up - regulated in high - dose FK506 treated group,was also markedly down- regulated in mice combined with Ala- Gln- treatment. A significantly increased expression of iNOS and TNF - α protein was found in the high - dose FK506 treated mice, while small amounts of these proteins were identified in the Ala - Gln - treated group. CONCLUSION: FK506 could induce a significant impairment of intestinal mucosa morphologically, which might be associated with up - regulated expression of iNOS and TNF - α in small intestinal mucosa. Subsequently, the intestinal permeability is increased. Ala - Gln has a strong protective effect on FK506 - induced intestinal barrier dysfunction, probably relates to the down - regulation of iNOS and TNF - α expression.