Objective To review our experience in the diagnosis and management of paralysis of the right hemidiaphragm after liver transplantation. Methods 60 adult patients received liver transplantation from February 2001 to March 2007 in Sun Yat-sen Memorial Hospital were retrospectively analyzed. The pathophysiologic changes, clinical progress, and management of serious respiratory complications caused by post-transplant paralysis of the right hemidiaphragm were studied. Results Among 60 patients, 40 developed postoperative respiratory complications, and 5 were due to paralysis of the right hemidiaphragm. The 5 patients presented with paradoxical respiration and the ventilator supporting times were 14, 16, 34, 45, and 60 days, respectively. Tracheostomy was performed in 4. These patients developed pneumonia in 5, atelectasis in 4, acute respiratory distress syndrome (ARDS) in 4, hepatopulmonary syndrome in 4, and pulmonay interstitial edema in 3. Among the 5 patients, 4 patients survived and 1 patient died of ARDS and multiple organs failure 31 days after the transplantation. Conclusions After liver transplantation, strict monitoring of the respiratory function and timely use of a respirator for patients with the paralysis of the hemidiaphragm is very important. For patients with suspicious hemidiaphragm paralysis, tracheostomy should be decisively performed.

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.

BACKGROUND: The immature dendritic cell (imDC) can induce immunological tolerance and has widely application in the field of organ transplant. At present, the methods of inducing imDC are insufficient, so the new induction method is demanding.OBJECTIVE: To investigate the effect of sodium butyrate (SB) on the maturation and immunological function of human peripheral blood-derived imDC.DESIGN: Controlled observation and in vitro cytological trial.SETTING: Department of Hepatobiliary Surgery in the Second Affiliated Hospital of Sun Yat-sen University.MATERIALS: Five samples of human peripheral blood were obtained from the healthy volunteers (aged 20-23 years) of Sun Yat-sen University, totally 500 mL. Then peripheral blood mononuclear cells (PBMCs) and lymphocytes were isolated within 2 hours.METHODS: The experiment was carried out in the Medical Research Center of the Second Hospital Affiliated to Sun (1 mmol/L) was added for induction, while those supplemented with maturation promoting factor lipopolysaccharide (LPS)the beginning of induction, while LPS was added on the sixth day for second stimulation.MAIN OUTCOME MEASURES: Cell morphological change, flow cytometry was used to detect DC phenotype,FITC-labeled Dextran was used to detect the endocytosis of DC, the production of IL-12 was determined by means of enzyme-linked immunosorbent assay, and the proliferation of lymphocyte induced by DC was assayed with mixed lymphocyte reaction.expressions of CD80, CD83 and HLA-DR were significantly lower in the imDC of routine induction group following SB maturity promoting, compared with LPS group (P＜0.01). On the sixth day, LPS was added into the SB-induced imDC,Endocytosis of DC: The imDC of routine induction group possessed a significantly lower endocytic activity after induced by LPS, and there were extremely significant differences compared with blank control group and SB maturation Production of IL-12: The production of IL-12 in the mDC induced by LPS was significantly higher than that in control group, SB maturation promoting group and SB induction group, the mDC induced by LPS in routine induction group stimulated significantly stronger proliferation of lymphocyte (P＜0.01).

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.

OBJECTIVETo investigate the gene expression of 4-1BB in peripheral blood mononuclear cells (PBMCs) and the possible significance of the 4-1BB pathway after clinical orthotopic liver transplantation (OLT).

METHODS4-1BB mRNA levels in PBMCs from 22 OLT patients were analyzed by RT-PCR. 4-1BB protein expressed on the surface of CD(4)(+) and CD(8)(+) T cells were detected by flow cytometry, and visualized with direct immunofluorescence and confocal fluorescence microscopy. Patients with primary liver cancer (PLC) and healthy volunteers served as controls. Six cases of recently performed liver transplantation were also observed in this study.

RESULTS4-1BB mRNA was detected in PBMCs from both liver transplant patients with long-term graft acceptance (22 cases) and from transplant patients on day 1 to day 3 post-transplantation (6 cases), but was not found in PBMCs from transplant patients on day 7 to day 30 post-transplantation (6 cases). 4-1BB mRNA was also not found in samples from 8 of the healthy controls and 7 of the PLC patients, though very low expression was detected in the other 4 healthy volunteers and 6 PLC patients. Simultaneously, 4-1BB protein was expressed at nearly undetectable levels on CD(4)(+) and CD(8)(+) T cells from healthy controls, PLC patients, as well as OLT patients within the first month post-transplantation (6 cases). However, 4-1BB expression was found on the surface of CD(4)(+) and CD(8)(+) T cells from liver transplant patients with long-term graft acceptance. Direct immunofluorescent staining and confocal fluorescence microscopy clearly revealed evidence of 4-1BB protein on cell membranes of CD(4)(+) and CD(8)(+) T cells from liver transplant patients with long-term graft acceptance. Simultaneously, a significantly higher percentage of CD(3)(+) CD(25)(+) T cells were found in liver transplant patients with long-term graft acceptance group as compared with the healthy control group (P < 0.05). The expression of 4-1BB protein on T cells did not correlate with the survival time of OLT patients postoperation.

CONCLUSIONSThis study demonstrates that although patients remain in stable condition after liver transplantation under the treatment of immunosuppressants, activated T cells are present to some extent and 4-1BB protein may be involved in this process. Effector T-cells can exert permanent immunoresponses against grafts under these circumstances. Therefore, we conclude that a new immune response balance is established under the combination of both treatment with immunosuppressants and natural immune responses against alloantigens. Manipulation of the 4-1BB/4-1BBL pathway may provide a therapeutic technique for prolonging graft survival.

Adult ; Antigens, CD ; Female ; Gene Expression ; Humans ; Leukocytes, Mononuclear ; chemistry ; Liver Transplantation ; Male ; Middle Aged ; Receptors, Nerve Growth Factor ; genetics ; physiology ; Receptors, Tumor Necrosis Factor ; genetics ; physiology ; Tumor Necrosis Factor Receptor Superfamily, Member 9

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. [

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.

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.

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

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