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Objective: To explore the characteristics of NF-KappaB activation in the progress of pancreatitis, the relationship with expression of TNF-alpha in the inflammatory reaction, and prevent the exacerbation of pancreatitis by using NAC., Method: Forty-eight rats were divided into three groups: therapy (group C), pancreatitis (group B) and control (group A). NAC served as the inhibitor of NF-KappaB activation. In the time intervals of 1.5, 3.0, 6.0, 12.0 hour, NF-KappaB activation was detected with flow cytometry (FCM) and the expression of TNF-alpha mRNA and protein with in situ hybridization (ISH) and enzyme-linked immuno-sorbent assay (ELISA) respectively. Meanwhile, the level of lipase and amylase in the serum was assayed and the pathological change was evaluated., Result: NF-KappaB activation in the pancreatitis group was higher than that in the control group (P<0.01), peaked at 3 hours, and was depressed by the inhibitor of NF-KappaB, NAC. The expression of TNF-alpha as well as the level of lipase and amylase in the serum also rose synchronously with activation of NF-KappaB. In contrast to group A, it was significantly different (P<0.01) in group B. After using NAC in group C, all of these values were decreased and the inflammatory reaction in the pancreas abated evidently. The pathology changes of the pancreas were shown to be alleviated in group C., Conclusion: First, NF-KappaB activity is intensively initiated in the course of pancreatitis and shown to have closely relationship with the release of cytokines. Second, use of NAC markedly depressed NF-KappaB activation. TNF-alpha expression is down regulated by cytokines. It is suggested that NAC probably acts as a useful agent for treatment of pancreatitis by indirectly inhibiting activation of NF-KappaB.

Background: We investigated the role of 1,25-dihydroxyvitamin D3(1,25-(OH)2D3) in preventing allograft from acute rejection following orthotopic liver transplantation., Methods: A rat orthotopic liver transplantation model was used in this study. SD-Wistar rats served as a high responder strain combination. Recipients were subjected to administration of 1,25-(OH)2D3 at dosages ranging from 0.25 microg.kg(-1).d(-1) to 2.5 microg.kg(-1).d(-1). Survival after transplantation as well as pathological rejection grades and IFN-gamma mRNA, IL-10 mRNA transcription intragraft on day 7, and day 30 post-transplantation were observed., Results: After recipients were treated with 1,25(OH)2D3 at dosages of 0.5 microg.kg(-1).d(-1) or 1.0 microg.kg(-1).d(-1), survivals of recipients were prolonged. Ninety-five percent confidence intervals of survival were 46 - 87 days and 69 - 102 days (both P = 0.0005 vs control group), respectively. On day seven post-transplantation, relative levels of IFN-gamma mRNA transcription were 0.59 +/- 0.12 and 0.49 +/- 0.16, which was higher than the control group (P = 0.005, P = 0.003, respectively). Relative levels of IL-10 mRNA transcription were 0.83 +/- 0.09 and 0.76 +/- 0.09, which was lower than the control group (P = 0.002, P = 0.003, respectively). At a dosage of 0.5 microg.kg(-1).d(-1), the median of pathological rejection grade on day seven and on day thirty post-transplantation were 1.5 and 2.0 in comparison with the CsA-treated group (P = 0.178, P = 0.171, respectively). At a dosage of 0.5 microg.kg(-1).d(-1), the median of pathological rejection grade on day seven and day thirty post-transplantation were 1.5 and 1.5 in comparison with CsA-treated group (P = 0.350, P = 0.693, respectively)., Conclusion: After each recipient was treated with 1,25-(OH)2D3 at a dosage of (0.5 - 1.0) microg.kg(-1).d(-1), transcription of cytokine intragraft was accommodated effectively and deviated to Th2 type, resulting in alleviation of acute rejection. 1,25-(OH)2D3 can prolong survival of recipient after orthotopic liver transplantation.

Objective: To identify the causes of respiratory complications following liver transplantation (LT) and to discuss the management of these complications., Methods: One hundred and twenty four cases with pulmonary complications in the first two weeks after LT were identified among 163 patients admitted to the First Affiliated Hospital, College of Medicine, Zhejiang University from February, 1999 to March, 2003., Results: The incidence rate of complications was 76%(124/163) with the total cure rate of 92%(114/124). The cure rates of the various complications were as follows: pleural effusion 100%(113/113), pneumonia 92%(76/83), respiratory insufficiency 91%(59/65), pulmonary hypertension 98%(101/103), pulmonary edema 98(58/59), atelectasis 100%(4/4) and pneumothorax 100%(2/2)., Conclusion: To drainage the pleural effusion with an unicameral venous catheter is safety and effective. To cure or prevent pneumonia and atelectasis, aseptic manipulating, aspiration of sputum and keeping respiratory channel open were the key measurements of treatment. Restrictive ventilatory functional disturbance (RVFD) and dysfunction of ventilation are two major types of respiratory insufficiency in early stage of post-transplantation. The causes of pulmonary hypertension and edema are associated with pulmonary angiotasis and blood flow volume, and the vasodilator and diuretic often introduced in the therapy.

Objective: To elucidate the profile of serum cytokines and adhesion molecules in stable survivors with clinical liver transplantation., Methods: Flow cytometric analysis was used to analyse the phenotype of T cell subsets in peripheral blood mononuclear cells (PBMCs) from group of liver transplantation (LTx) (n = 22), primary liver carcinoma (PLC) (n = 13) and healthy control (n = 12). Enzyme-linked immunoabsorbent assay (ELISA) was used to determine the serum cytokines and adhesion molecules profiles in stable survivors with clinical liver transplantation., Results: Percentage of CD3(+) T cell and CD8(+) T cell, as well as ratio of CD4(+) to CD8(+) revealed no difference among three groups. The percentage of CD3(+)CD25(+) T cells in LTx group was found higher than that in healthy group (P = 0.022). Th1 cytokines (IL-2, IFN-gamma) and Th2 cytokines (IL-4, IL-10), as well as TNF-alpha displayed no significant difference among three groups. The levels of IL-6, ICAM-1 and P-selectin in serum were not found any difference between LTx group and PLC group, while the levels of IL-6, ICAM-1 and P-selectin in serum shown significant difference between LTx and healthy groups (P = 0.048, 0.000 and 0.025, respectively)., Conclusions: Our data demonstrates that effector T-cells can also be activated and exert immunoresponse to grafts permanently under the treatment of immunosuppressant. Adhesion molecules (ICAM-1, P-Selectin) and pro-inflammatory cytokines (IL-6, TNF-alpha) might be involved in the process of chronic graft damage induced by allo-immunoresponse.

Background: Hepatic artery thrombosis (HAT) which is a serious complication after orthotopic liver transplantation (OLT) remains a significant cause of graft loss. The purpose of this study was to sum up our experiences in the prevention, diagnosis and management of HAT after liver transplantation., Methods: From April 1993 to September 2003, a total of 198 patients underwent OLT at our hospital. The hepatic artery was anastomosed using 7/0 prolane with running continuous suture in 96 patients (group 1) and with interrupted suture in 102 (group 2). Ultrasonography was performed every day in two weeks after operation and selectively afterwards., Results: HAT occurred in 6 patients (6.3%, 6/96) of group 1, and in 1 (1%, 1/102) of group 2 (X2=4.027, P=0.045). Six patients received emergency thrombectomy, and 1 conservative therapy but died from tumor recurrence eventually. Biliary complication developed in 3 patients after thrombectomy of whom 2 died of liver failure and one waited for retransplantation. In the other 3 patients after thrombectomy, 1 died of renal failure, and 2 survived. The mortality of patients with HAT was 57.1% (4/7)., Conclusions: The technique of hepatic arterial anastomosis is the key factor for the prevention of HAT. Routine ultrasonography is very important in early detection of HAT after OLT. Biliary complication is a severe outcome secondary to HAT.

In order to study the molecular mechanism of the inhibitory effect of 1,25-dihydroxyvitamin D(3) on dendritic cells, experiments were performed using Atlas cDNA expression arrays from Clonetech to identify the differentially expressed genes of dendritic cells by 1,25-dihydroxyvitamin D(3). Analysis of cDNA arrays revealed changes in the expression of 9 genes, including those involved in DNA binding and transcription, extracellular cell signaling and communication, intracellular transducers, as well as cell adhesions. The results indicated that a multiple molecular network is involved in the inhibitory role of 1,25-dihydroxyvitamin D(3) on dendritic cells. The Atlas Array technology may facilitate the elucidation of complex pharmacological process of 1,25-dihydroxyvitamin D(3) on dendritic cells.

To evaluate the effects of proteasome inhibitors lactacystin (LAC) and beta-lactacystin (beta-LAC) on the proliferation and activation of T lymphocytes, flow cytometry was used to analyze the proliferation and the expression of CD69, CD25 and CD3 of T lymphocytes activated by PHA. Furthermore, the expressions of PA28 and IL-2 mRNA were assayed by competitive RT-PCR. The results indicated that: (1) LAC and beta-LAC significantly decreased the incorporation of BrdU and inhibited T lymphocytes proliferation in T lymphocytes activated by PHA; (2) although LAC and beta-LAC did not affect the expression of CD69 at any time, they significantly inhibited the expression of CD25 (48 h, 72 h, P<0.05); (3) in comparison with control, LAC and beta--LAC significantly down-regulated the expression of PA28 and IL-2 mRNA (48 h, 72 h, P<0.05). LAC and beta-LAC significantly inhibited the proliferation and activation of T cells. Mechanisms involved are inhibition of CD25 and down-regulation of PA28 and IL-2 mRNA expressions.

Aim: To investigate the expression of 4-1BB molecule in hepatocellular carcinoma (HCC) and its adjacent tissues., Methods: Reverse transcription-polymerase chain reaction (RT-PCR) was used to determine the gene expression of 4-1BB in hepatocarcinoma and its adjacent tissues, and peripheral blood mononuclear cells (PBMCs) from both HCC and health control groups. Flow cytometry was used to analyse the phenotypes of T cell subsets from the blood of HCC patients and healthy volunteers, and further to determine whether 4-1BB molecules were also expressed on the surface of CD4+ and CD8+ T cells. The localization of 4-1BB proteins on tumor infiltrating T cells was determined by direct immunofluorescence cytochemical staining and detected by confocal microscopy., Results: 4-1BB mRNA, which was not detectable in normal liver, was found in 19 liver tissues adjacent to tumor edge (<1.0 cm). Low expression of 4-1BB mRNA was shown in 8 tumor tissues and 6 liver tissues located within 1 to 5 cm away from tumor edge. In PBMCs, 4-1BB mRNA was almost not detected. Percentage of CD4+, CD8+ and CD3+/CD25+ T cells, as well as ratio of CD4 to CD8 revealed no difference between groups (P>0.05, respectively), while a significant lower percentage of CD3+ T cell was found in HCC group as compared to healthy control group (P<0.05). However, 4-1BB molecules were almost not found on the surface of CD4+ and CD8+ T cells in HCC and healthy control group. Double-staining of 4-1BB+/CD4+ and 4-1BB+/CD8+ immunofluorescence on tumor infiltrating T cells was detected in 13 liver tissues adjacent to tumor edge (<1.0 cm) by confocal microscopy., Conclusion: Although HCC may escape from immune attack by weak immunogenicity or downregulated expression of MHC-1 molecules on the tumor cell surface, tumor infiltrating T cells can be activated via other costimulatory signal pathways to exert a limited antitumor effect on local microenvironment. The present study also implicates that modulating 4-1BB/4-1BBL costimulatory pathway may be an effective immunotherapy strategy to augment the host response.

Aim: To investigate the inhibitory effect of tea polyphenols (TP) on the transforming growth factor-beta1 (TGF-beta1) expression in rat model of cyclosporine A (CsA)-induced chronic nephrotoxicity., Methods: The rat model of CsA-induced chronic nephrotoxicity was used, 4 groups of rats were respectively treated with vehicle (0.1 mL/kg/d sc), TP (80 mg/kg/d ig), CsA (15 mg/kg/d sc) and TP plus CsA (CsA 15 mg/kg/d sc+TP 80 mg/kg/d, ig). At the end of day 28 of treatment, serum and urine are analyzed for creatinine clearance, kidney tissue for pathologic analysis. The TGF-beta1 mRNA and its protein expression were detected by RT-PCR, immunohistochemistry, and Western blot., Results: CsA-treated rats had increased renal expression of TGF-beta1 mRNA and its protein, compared with the vehicle- or TP-treated controls. The renal function and interstitial fibrosis were ameliorated and renal expression of TGF-beta1 mRNA and its protein was decreased in animals treated with CsA plus TP, compared with animals treated with CsA alone (P<0.05)., Conclusion: TP significantly inhibits renal expression of TGF-beta1 in rat model of cyclosporine-induced chronic nephrotoxicity, suggesting that the decreased renal expression of TGF-beta1 exerted by TP is one of mechanisms to protect renal function and tissue structure.

Objective: To explore the effect of two dominating signaling pathways, VEGF/KDR and angiopoietins/Tie2, on the formation of new blood vessel in hepatocellular carcinoma (HCC) growth and metastasis., Methods: RT-PCR and Western blot were employed to evaluate the VEGF/KDR and angiopoietins/Tie2 expression in samples from 23 patients with HCC. Meanwhile, microvessel density (MVD) was determined as a marker of angiogenesis by counting CD34 positive cells with the method of immunohistochemistry., Results: The two pathways were activated in all HCC samples. The expressions of vascular endothelial growth factor (VEGF) and angiopoietin-2 (Ang2) were significantly higher (P<0.05) in hepatocellular carcinoma tissues and the margin of the tumor than those in control groups, and so did CD34 positive cells. Although significant difference in the expression of kinase insert domain containing receptor (KDR) and Ang1/Tie2 was not observed in all groups, their distinct high levels were seen in hepatoma and its margin compared with normal and cirrhotic liver. VEGF and Ang2 expressions were seen up-regulated in HCC with vascular invasion and satellite lesion., Conclusions: The two signaling pathways, VEGF/KDR and angiopoietins/Tie2 are activated in the process of angiogenesis in HCC and modulate the formation of new blood vessels. The imparity of the two signaling pathways' activation is to benefit HCC metastasis. In the two pathways, VEGF and Ang2 may play an important role in the process of angiogenesis, and are necessary indicators for the prognosis and metastasis of HCC. This study provides another clue for the exploration of anti-angiogenic agents.

Objective: To use 2.2.15 cell line to determine the effects of mycophenolate acid (MPA) on hepatitis B virus (HBV) replication and viral protein synthesis in vitro., Methods: The 2.2.15 cells were treated with different concentration of MPA (1-50 mug/ml) for 12 days. HBsAg and HBeAg were detected in the supernatant fluid by ELISA and intracellular HBV DNA was analyzed quantitatively by slot blot hybridization., Results: MPA could suppress the expression of HBsAg and HBeAg, and the higher concentration of MPA induced lower expression of HBsAg and HBeAg. The suppression rates of MPA for HBsAg and HBeAg at a concentration of 50 micrograms/ml were 34.2% and 24.1% respectively. The expression of HBV DNA was only 49% as compared with controls when treated with MPA at a concentration of 50 micrograms/ml., Conclusions: Mycophenolate acid can suppress the expression of HBsAg and HBeAg as well as the replication of HBV DNA in the 2.2.15 cell. The suppressive degree is dose-dependent.

Aim: To modify the hamster-to-rat liver xenotransplant technique to prevent postoperative complications, and to study the inhibiting effect of multiglycosides tripterygium wilfordii (T(II)) on immune rejection., Methods: Female golden hamsters and inbred male Wistar rats were used as donors and recipients, respectively. One hundred and twelve orthotopic liver xenotransplants were performed by Kamada's cuff technique with modifications. Over 72 hour survival of the animal after operation was considered as a successful operation. When the established surgical model became stable, 30 of the latest 42 cases were divided into untreated control group (n=15) and T(II) group (n=15) at random. Survival of recipients was observed. Liver specimens were collected at 2 and 72 h from the operated animals and postmortem, respectively, for histological study., Results: The successfully operative rate of the 30 operations was 80 %, and the survival of the control and T(II) group was 7.1+/-0.35 was days and 7.2+/-0.52 days, respectively (t=0.087,P=0.931). The rate of conjunctival hyperemia in control group (100 %) differed significantly from that (31 %) in T(II) group (P=0.001). Rejection did not occur in both groups within 2 h postoperatively, but became obvious in control group at 72 h after surgery and mild in T(II) group. Although rejections were obvious in both groups at death of recipients, it was less severe in T(II) group than in control group., Conclusion: This modified Kamada's technique can be used to establish a stable hamster-to-rat liver xenotransplant model. Monotherapy with multiglycosides tripteryguiumwilfordii (30 mg x kg(-1) x d(-1)) suppresses the rejection mildly, but fails to prolong survival of recipients.

Unlabelled: Use and effects of liver specific antigen in orthotopic liver transplantations were researched in this study. Group I: syngeneic control (Wistar-to-Wistar); Group II: acute rejection (SD-to-Wistar); Group III: Thymic inoculation of SD rat LSA day 7 before transplantation. The observation of common situation and survival time, rejection grades, NF-KappaB activity of splenocytes and IL-2mRNA expression of grafted liver were used to analyze acute rejection severity and immune state of animals in different groups. The common situation of group I was very well after transplantation and no signs of rejection were found. Recipients of group II lost body weight progressively. All dead within day 9 to day 13 posttransplantation; median survival time was 10.7+/-0.51 days. It was an optimal acute rejection control. As for group III, 5 out of 6 recipients survived for a long time and common situation was remarkably better than that of group II. Its rejection grades were significantly lower than that of group II(P < 0.05). NF-KappaB activity was only detected in group I at day 5 and day 7 after transplantation, whereas high activity of NF-KappaB was detected at all time points in group II and the low NF-KappaB activity detected in group III was significantly lower than that of group II(P < 0.05). No IL-2mRNA expression was detected at any time point in group I, whereas high level expression was detected at all time points in group II and the low level expression only detected at day 3 in group III was significantly lower than that of group II(P < 0.05)., Conclusion: LSA is an important transplantation antigen which is involved directly in the immunorejection of liver transplantation. We report here for the first time that intrathymic inoculation of LSA can alleviate the rejection of liver allotransplantation; and that grafts can survive for a long time thereby, thus leading to a novel way to achieve liver transplantation immunotolerance.

Objective: To investigate the effect of combined CsA and FK506 with 5-FU on hepatocellular carcinoma rats., Methods: A syngeneic rat model of hepatocellular carcinoma was used. Control group (A) underwent 4 ml 5% GS. Treatment group was divided into 3 groups namely, group B: only 5-FU and 5% GS; group C: 5-FU, CsA and 5% GS; group D: 5-FU, FK506 and 5%GS. Cell cycle, apoptosis, necrosis and mitochondrial transmembrane potential were measured by flow cytometry, laser scanning confocal microscopy, and electron transmission microscopy. Statistical analysis was performed by SPSS 10.0 for Windows software. Statistical comparisons were made with ANOVA followed by Dunnett's T3 or LSD test., Results: Compared to the control group, the percentage of apoptotic cells including trifle necrotic cells was significantly higher, and among the treatment group, group D was the highest, and group C was higher than group B. In the treatment group, cell cycle of hepatoma cells was mainly arrested at S phase, but in group D, G0/G1 phase cells were significantly decreased and S phase cells significantly increased. Compared to the control group, mitochondrial transmembrane potential was significantly decreased in the treatment group, among with, group B was the lowest, group C was higher than group D. Morphological changes demonstrated by electron microscopy included dispersed nuclear chromatin, loss of nucleoli, membrane bleeding, cell shrinkage, typical apoptotic bodies and marked swelling of mitochondria in the treatment group. In the control group, however, they were characterized by normal cell ultrastructure., Conclusions: The present study reveals that 5-FU combined with CsA or FK506 demonstrated a synergistic effect on hepatocellular carcinoma rats. For FK506, the powerful mutual effect is related to the increase of tumor cell's quantity in S phase. Both CsA and FK506 can provide protection on mitochondrial transmembrane potential reduction against hepatoma cells damage from 5-FU.

Aim: To study the mechanism and the preventive role of 1,25-dihydroxyvitamin D3 in acute rejection following orthotopic liver transplantation., Methods: Rats were randomly divided as donors or recipients for orthotopic liver allotransplantation model. Four groups were designed in the study, Group I: syngenic control (Wistar to Wistar); Group II: acute rejection (SD to Wistar); Group III: acute rejection treated with cyclosporine A, and Group IV: acute rejection treated with 1,25-(OH)2D3. Liver function, rejection activity index and mRNA of IFN-gamma, IL-10 in intragraft in recipients were measured in on day 1, 5, 7, 15, 30 posttransplant for assessing graft function, severity of acute rejection and immune state of recipients., Results: Survival time of recipients in Group VI was significantly prolonged (over 100 days) in comparison with other groups (vs Group II, P<0.001; vs Group III, P>0.05). After treatment with 1,25-(OH)2 D3, mean value of all the assay tested on each experimental time was compared, liver function in group IV was significantly improved (AST 127+/-41 U/L-360+/-104 U/L, BIL 13+/-5 mmol/l-38+/-11 mmol/l; vs Group II, P<0.05; vs Group III, P>0.05. Rejection activity index was significantly decreased (0-3.3+/-1.6; vs Group II, P<0.05; vs Group III, P>0.05). Level of hepatic IFN-gamma mRNA in group IV was decreased, while level of hepatic IL-10 mRNA was increased (vs Group II, P<0.05; vs Group III, P>0.05)., Conclusion: Our results indicated that 1,25-(OH)2D3 induced the secretion of cytokine toward to Th2 type, which would alleviate acute rejection, protect liver function and prolong survival of recipient after orthotopic liver transplantation.

Nuclear factor-kappa B (NF-kappa B) as an essential transcription factor in the control of expression of the cytokine-induced genes in immune and inflammatory responses regulates cytokines in allograft rejection. In this review, we summarize the general properties of NF-kappa B and the principal findings to shed new light on transplantation.

Objectives: To explore the relation between tumor TNM of pancreatic cancer and treatment, and to summarize the experience in diagnosis and treatment of pancreatic cancer., Methods: The data on 321 patients with pancreatic cancer treated at our hospital between January 1990 and December 2000 were reviewed retrospectively. Radical resection and survival for various TNM stages were analyzed., Results: The rates of radical resection for stage I, stage II, stage III, stage IVA, and stage IVB were 100%, 100%, 82.6%, 12.3% and 2.1% respectively. The 1-, 3- and 5-year survivals of stage I, II were 73.3%, 40.0% and 30.0% respectively, and higher than those of other groups., Conclusions: The patients with pancreatic cancer diagnosed early can undergo radical resection. The elevation of early diagnosis rate is significant for the improvement of survival.

Objective: To investigate the alloimmunogenicity of liver specific antigen and its effects on allolymphocytes., Methods: Liver specific antigen isolated from inbred F344 rats was used as immunogen to immunize inbred Lew rats through different immunization pathways such as low-dose long-term hind footpad, high-dose portal vein and thymus immunization. Western blotting, DNA fragments gel electrophoresis, mixed lymphocyte culture (MLC) and mixed lymphocyte hepatocyte culture (MLHC) were employed to analyze the immune state after immunization., Results: At the time point of sampling, different degree of specific low immunoresponses appeared in all immunized groups as well as cyclophosphamide (CY) treated group. Compared with group I, other groups expressed caspase-3 significantly as detected by using Western blotting. DNA fragment gel electrophoresis of splenocytes showed lymphocyte apoptosis. Compared with the group I, MLC of the experimental groups showed no significant changes except that of the group V, whereas MLHC decreased markedly (P<0.05)., Conclusions: Liver specific antigen not only has alloimmunogenicity to induce alloimmunoreaction but induce antigen specific low immunoresponses and antigen specific lymphocyte apoptosis by high-dose or low-dose long-term immunization. It may be an important transplantation antigen that may lead to a novel way to liver transplantation immunotolerance.

Objective: To investigate the expression of tumor necrosis factor-alpha(TNF-alpha), interleukin-1 beta (IL-1beta), and intercellular adhesion molecule-1 (ICAM-1) in the lung of rat with small-for-size liver transplantation and the significance of the expression of these cytokine in lung injury after liver transplantation., Methods: 150 SD rats were randomly divided into 4 groups: sham operation group (n = 6), whole graft liver transplantation group (n = 48), 50% size graft liver transplantation group (n = 48), and 30% size liver transplantation group (n = 48). Six rats from each group were killed 0.5, 2, 6, and 24 hours after operation. Blood samples from subhepatic inferior vena cava were obtained to examine the plasma TNF-alpha by ELISA. Specimens of lung were obtained to be examined pathologically. RT-PCR was used to examine the expression of TNF-alpha mRNA, IL-1beta mRNA, and ICAM-1 mRNA in lung, and chromatometry was performed to detect the activity of myeloperoxidase (MPO)., Results: (1) The plasma TNF-alpha at any time point was higher in the 3 transplantation groups than in the sham operation group. The plasma TNF-alpha 2 hours after operation in the whole graft group was significantly higher than that in the 30% size group (P < 0.05). (2) The expression levels of TNF-alpha mRNA 2 and 6 hours after operation in the whole graft group and in the 50% graft group were significantly higher than those in the 30% graft group (all P < 0.01). The expression levels of TNF-alpha mRNA remained significantly higher than those in the sham operation group since the second hour after operation (all P < 0.01). (3) IL-1beta mRNA was expressed in the 3 liver transplantation groups without significant differences between any levels at all the time points and was not expressed in the sham operation group. The expression of ICAM-1 mRNA was higher at all the time points in the liver transplantation groups than in the sham operation group (P < 0.01 or P < 0.05) without significant difference between the values of any transplantation group at any time point (all P > 0.05). The MPO activity was stronger in the 3 liver transplantation groups at any time point than in the sham operation group (all P < 0.01). The peak occurred 2 hours after operation in the whole graft group and 50% size group and occurred 6 hours after operation in the 30% graft group. The plasma TNF-alpha level was positively correlated to the MPO activity in lung tissue with a correlation coefficient of 0.422 (P < 0.05). (4) The morphology of lung was normal in the sham operation group. Obvious interstitial hyperemia and hemorrhage, neutrophil infiltration, and pulmonary septal thickening were found in the 3 transplantation groups, in particular being severe at the 2-hour time point., Conclusion: Increase of plasma TNF-alpha is one of the causes of lung injury after small-for-size liver transplantation. Upregulation of TNF-alpha mRNA, IL-1beta mRNA, and ICAM-1 mRNA expression may be also responsible for the lung injury and their expression may be correlated to the size of graft.

Aim: To study the effects of liver specific antigen (LSA) on the immunoreaction of liver allotransplantation and its significance., Methods: Orthotopic liver transplantation was used in this study. Group I: syngeneic control (Wistar-to-Wistar); Group II: acute rejection (SD-to-Wistar). Group III: acute rejection treated by intramuscular injection of cyclosporine A (CsA) (SD-to-Wistar+CsA). Group IV: Intrathymic inoculation of SD rat LSA one week before transplantation (LSA+SD-to-Wistar). The common situation and survival time, rejection grades, NF-kappaB activity of splenocytes and intragraft cytokine gene expression were observed to analyze the acute rejection severity and immune state of animals., Results: The common situation of Wistar-to-Wistar group was very good after the transplantation and no signs of rejection were found. Recipients of SD-to-Wistar group lost body weight progressively. All died within 9 to 13 days after transplantation with the median survival time of 10.7+/-0.51 days. It was an optimal control for acute rejection. The common situation of SD-to-Wistar+CsA group was bad during CsA medication but only with mild rejection. As for LSA+SD-to-Wistar group, 5 of 6 recipients survived for a long time and common situation was remarkably better than that of SD-to-Wistar group and SD-to-Wistar+CsA group. Its rejection grades were significantly lower than that of SD-to-Wistar group (P=0.026). Furthermore, no significant discrepancies of rejection were found between SD-to-Wistar group and LSA+SD-to-Wistar group at day7 and day12 (P=0.067). NF-kappaB activity, IFN-gamma and IL-2mRNA expression were significantly inhibited in LSA+SD-to-Wistar group compared with that of SD-to-Wistar group (P<0.05)., Conclusion: LSA is an important transplantation antigen which involves in the immunorejection of liver transplantation directly. We reported for the first time that intrathymic inoculation of LSA can induce immnotolerance of liver allotransplantation and grafts can survive for a long time thereby, thus leading to a novel way to liver transplantation immunotolerance.

Aim: To investigate the expression of growth hormone receptor (GHR) and mRNA of GHR in cirrhotic livers of rats with the intension to find the basis for application of recombinant human growth hormone (rhGH) to patients with liver cirrhosis., Methods: Hepatic cirrhosis was induced in Sprague-Dawley rats by administration of thioacetamide intraperitoneally for 9-12 weeks. Collagenase IV was perfused in situ for isolation of hepatocytes. The expression of GHR and its mRNA in cirrhotic livers was studied with radio-ligand binding assay, RT-PCR and digital image analysis., Results: One class of specific growth hormone-binding site, GHR, was detected in hepatocytes and hepatic tissue of cirrhotic livers. The binding capacity of GHR (R(T), fmol/mg protein) in rat cirrhotic liver tissue (30.8+/-1.9) was significantly lower than that in normal control (74.9+/-3.9) at the time point of the ninth week after initiation of induction of cirrhosis (n=10, P<0.05), and it decreased gradually along with the accumulation of collagen in the process of formation and development of liver cirrhosis (P<0.05). The number of binding sites (X10(4)/cell) of GHR on rat cirrhotic hepatocytes (0.86+/-0.16) was significantly lower than that (1.28+/-0.24) in control (n=10, P<0.05). The binding affinity of GHR among liver tissue, hepatocytes of various groups had no significant difference (P>0.05). The expression of GHR mRNA (riOD, pixel) in rat cirrhotic hepatic tissues (23.3+/-3.1) was also significantly lower than that (29.3+/-3.4) in normal control (n=10, P<0.05)., Conclusion: The growth hormone receptor was expressed in a reduced level in liver tissue of cirrhotic rats, and lesser expression of growth hormone receptors was found in a later stage of cirrhosis. The reduced expression of growth hormone receptor was partly due to its decreased expression on cirrhotic hepatocytes and the reduced expression of its mRNA in cirrhotic liver tissue.

Background: The molecular mechanism of small-for-size graft injury remains unclear. The aim of this study is to investigate the gene expression pattern of acute phase response in relation to graft size in a rat-liver transplantation model., Methods: A rat orthotopic liver transplantation model using 30%, 50%, and whole grafts was used. The graft survival rates and liver morphology were compared among the three groups. Two transcription factors, nuclear factor (NF)-kappaB (p65) and early growth response (Egr-1), and their downstream genes were compared., Results: According to the graft size, the rats were grouped as follows: group 1 (n=20), 32% (24-47%); group 2 (n=10), 56% (50-65%); and group 3 (n=10), 104% (89-120%). The 7-day survival rates were 20% (P=0.039 vs. group 2, P=0.000 vs. group 3), 60%, and 100% in groups 1, 2, and 3, respectively. Dilation of hepatic sinusoids and vacuolization of hepatocytes were observed in group 1. Up-regulation of Egr-1 and endothelin (ET)-1 and over-expression of nitric oxide synthase (iNOS) was found in group 1, but heme oxygenase (HO)-1 and A20 were down-regulated. At 24 hours after reperfusion, the intragraft protein level of heat-shock protein (Hsp)-70 was significantly lower in group 1 than that in group 3 (12.4 vs. 17.0 ng/mL, P=0.04). More apoptotic nuclei were found in group 1., Conclusions: Small-for-size graft injury was related to early over-expression of Egr-1 associated with up-regulation of ET-1 and deterioration of intracellular homeostasis reflected by down-regulation of Hsps and A20.

Objective: To investigate the gene expression of 4-1BB in peripheral blood mononuclear cells (PBMCs) and its possible significance in clinical liver transplantation., Methods: Reverse transcription-polymerase chain reaction (RT-PCR) was used to determine the gene expression of 4-1BB in PBMCs from 22 patients receiving liver transplantation, 13 patients with primary liver carcinoma (PLC), and 12 healthy controls. To determine whether 4-1BB molecule is also expressed on the surface of CD4+ and CD8+ T cell, flow cytometry was used to analyse the phenotype of T cell subsets from the blood of liver transplantation patients., Results: 4-1BB mRNA was detected in PBMCs from stable survivors after liver transplantation, but almost not detected in PBMCs from PLC patients and healthy controls. Meanwhile, 4-1BB was almost not expressed on the surface of CD4+ and CD8+T cells in healthy controls and PLC patients. A low level of 4-1BB expression, however, was found on the surface of CD4+ and CD8+ T cells from the stable survivors after liver transplantation., Conclusions: This study demonstrates that although patients are stable after liver transplantation, effector T-cells can also be activated through the signal of 4-1BB molecule and persistent immune response to grafts. Blockage of 4-1BB/4-1BBL pathway may benefitially reduce the clinical dosage of immunosuppressive agents and prolong the survival of grafts.

Objectives: To understand the characteristics of primary liver cancer (PLC) in the elderly and summarize the experience in treatment of such patients., Methods: The clinical data of PLC in the elderly group (>/=60 years, 125 patients) and the young group (0.05)., Conclusions: Hepatectomy is a choice of treatment for PLC in the elderly based on their liver function. AFP and B-ultrasonography are important methods for the diagnosis of PLC in the elderly.

Objective: To summarize our clinical experience in liver transplantation while considering the background in this field in China., Methods: Ninety-five patients who had received liver transplantation from April 1993 to March 2002 were analyzed retrospectively. Three periods were defined objectively as period I (1993 - 1997), II (1999) and III (2000 - 2002). Operative techniques, recipients, original diseases, complications and survival rates were compared among the three periods., Results: Malignant liver lesions were the main cause for liver transplantation in period I and II. The ratio of number of malignant disease to total recipients decreased gradually from period I to III (100%, 53% and 35%, respectively). The 1-year survival rate in patients with benign liver disease was 85% and the total operative mortality was 5% in period III. The incidence of hepatitis B virus reactivation or reinfection was 24% twelve months after liver transplantation. Vascular complication decreased but biliary complications did not and remained a major long-standing problem. No veno-venous bypass technique was used in period III, and its advantages were obvious when comparing with those with veno-venous bypass in period I and II., Conclusions: Strict selection of recipients, fine operative technique, familiarity with various complications and correct therapeutic methods, prophylaxis of recurrence of hepatitis B and hepatocellular carcinoma are necessary to improve long-term results of liver transplantation in China.

Objective: To study the real effect of IL-15, a kind of T lymphocyte activators which were derived from lymphocytes, on the acute rejection process in heart and liver transplantation in rats., Methods: Male (body weight 200 - 250 g) 1A (RT1(a)) and LEW (RT1(l)) rats were selected as donors and recipients, respectively. Heterotopic heart transplantation (in abdomen) and orthotopic liver transplantation were performed as the acute rejection model group (experimental group); LEW (RT1(l))-->LEW (RT1(l)) as donors and recipients to establish isografts transplantation as the control group. Animals were sacrificed on day 1, 3, 5, 7 and graft specimens were collected. Microarray, immunohistochemistry and Western-blotting methods were used to detect the expressions of IL-15, IL-2 and IFN-gamma, etc. 48 rats were divided evenly into two groups and each time-point consisted of 6 rats., Results: Acute rejections which were clarified by pathological findings and animal manifestations were found 3 days after operation in the experimental group. The early expression of IL-15 was found on endothelial cells in allografts 1 day after operation in contrast to IL-2, which expressed lately and only be found on inflammatory cells including lymphocytes and Kupffer cells 3 days after graft implantation. The result of INF-gamma was the same as that of IL-2., Conclusions: IL-15 appeared earlier in heart and liver allografts than IL-2 and IFN-gamma in rat acute rejection model, and the expression site differed from the later two. IL-15 participated in acute rejection reaction earlier in this process and the pathway may be different from IL-2 and IFN-gamma. Early blocking this pathway combined with other blockade would have a promising result in control of the progression of acute rejection.

Objective: To sum up the experience in liver transplantation in a period of ten years at a single center., Methods: We retrospectively reviewed the clinical records of 120 patients receiving liver transplantation from April 1993 to October 2002. The patients' clinical characteristics, surgical techniques, complications and survival were compared in the phases of 1993-1997 (phase I), 1999 (phase II), and 2000-2002 (phase III)., Results: Malignant liver diseases were major indications for liver transplantation in phase I (100%) and II (53.3%), but decreased markedly in percentage in phase III (34.0%). When compared with recipients in phase I and II, the survival of recipients with benign liver diseases in phase III was significantly improved with the 3-month, 6-month and 1-year survival rates of 85.7%, 84.5% and 83.1%, respectively. For patients with malignant liver diseases, the 3-month, 6-month and 1-year survival rates were 87.4%, 81.1% and 46.0%, respectively. The reinfection rate of hepatitis B virus was 24% 12 months after transplantation. With technical refinements, the incidence of postransplantation vascular complications has significantly decreased from 29.4% in phase I and II to 4.9% in phase III. Biliary complications remained one of the major obstacles to long-term survival. No veno-venous bypass was applied in phase III, providing a promising outcome., Conclusion: Strict selection of potential recipients, technical refinement, appropriate management of vascular and biliary complications, and prophylaxis of recurrences of hepatitis B and malignant liver diseases are important to obtain long-term survival of patients receiving liver transplantation in China.

Objectives: To prevent and early diagnose hepatitis B virus reinfection and recurrent hepatitis B following liver transplantation, and to discuss the further treatment of recurrent hepatitis B., Methods: Liver transplantation recipients received lamivudine for prophylaxis of HBV reinfection. Virological and biochemical data, serum HBV DNA, and immunohistological staining for HBsAg and HBcAg in liver biopsy specimens were tested in due time., Results: Five patients with hepatitis B virus reinfection and two patients with hepatitis B recurrence were observed after liver transplantation. One patient with recurrent hepatitis B developed chronic severe hepatitis B despite treatment. One patient improved after a series of treatment., Conclusions: Hepatitis B virus reinfection or recurrent hepatitis B following liver transplantation occurs mostly 6-12 months after operation. The diagnosis of hepatitis B recurrence should be taken into account when liver biochemical data becomes poor during this period. The treatment for recurrent hepatitis B after liver transplantation includes increased dosage of lamivudine, application of famciclovir, and other liver protection measures.

Objective: To investigate the variation of functions of the heart and lung during orthotopic liver transplantation (OLT)., Methods: Pulmonary artery cannula and right radial artery cannula were indewelled before anaesthesia and the parameters of hemodynamics in different periods were monitored. Analysis of variance was used to reveal the variation among the groups. T hypothesis test in paired data was used to compare the preoperative parameters with those in each period during operation and after operation respectively, and to compare the parameters immediately after operation with those in each period after operation respectively., Results: During the operation, heart rate increased, but blood pressure decreased significantly at the beginning of no-liver period, increased again in a short period and then increased progressively 12 h after operation. Pulmonary artery pressure (PAP) increased from before the no-liver period to 60 h after operation. Pulmonary wedge pressure changed in accordance with the variation of PAP. Cardiac output was maintained at a high-output level from before operation to 60 h after operation. Systemic vascular resistance (SVR) was within the normal limits before operation, whereas pulmonary vascular resistance (PVR) was lower than normal. In the no-liver period during the operation, SVR decreased significantly. Both SVR and PVR increased progressively and returned to normal postoperatively., Conclusions: The patients undergoing OLT have a high cardiac output and low resistance obstacle before and during the operation, and will recover gradually after operation. Monitoring hemodynamics during the peri-operative period is of significance in the prevention and treatment of pneumonedema and cardiac functional insufficiency.

Objective: To study the relationship between the serum levels of hyaluronic acid (HA), procollagen type III (PCIII), collagen type IV (CIV) and the histological degree of hepatic fibrosis evaluated by image analysis, and the clinical significance of serum HA, PCIII, CIV in the diagnosis of hepatic fibrosis in patients with chronic viral hepatitis., Methods: The concentrations of serum HA, PCIII, CIV in 151 patients with chronic viral hepatitis were measured by radioimmunoassay. Liver biopsies were performed in all the patients. Histological sections of 4 microm thickness were stained with Masson's trichrome for fibrosis assessment. Morphometric quantitative measurements for hepatic fibrosis assessment in the 4 microm sections were performed using a fully automated image analysis system. Serum levels of HA, PCIII, and CIV were analyzed at different stages of liver pathology and compared with the morphometric quantitative measurements of hepatic fibrosis., Results: The serum levels of HA, PCIII, CIV all elevated gradually with the progression of the disease, and all reached the highest in patients with liver cirrhosis. There was a significant difference in the levels of these 3 components between liver cirrhosis group and the other groups (P<0.05). They all increased steadily with the histological stages of hepatic fibrosis, and reached the highest levels in stage IV. The serum levels of HA, PCIII, CIV were all positively correlated with the histological stages of liver sections and the morphometric measurement (P<0.001). The coefficients with stages were 0.694, 0.493, 0.552 (P<0.001), respectively and with surface density of total collagen on liver biopsy sections by image analysis were 0.715, 0.595, 0.573 (P<0.001), respectively., Conclusion: The serum levels of HA, PCIII, CIV were in consistent with the degree of hepatic fibrosis, and the determination of these marks is valuable for detecting hepatic fibrosis.

Objective: To sum up the preliminary experience in living related liver transplantation (LRLT)., Methods: A 9-month-old male infant with biliary atresia (BA) who had undergone an unsuccessful Kasai operation was defined as a candidate for LRLT. The donor was his 30-year-old mother. Her lateral lobe of the left liver was transplanted into the infant's body as the graft. The left branches of the portal vein, left hepatic artery and left hepatic vein of the graft were end-to-end anastomosed to the portal vein, hepatic artery proper and hepatic vein of the recipient respectively. Biliary drainage was reestablished via Roux-en-Y operation., Results: The donor retained her liver function within 2 weeks after the operation. Steroid and FK506 were prescribed in immunosuppressive therapy for the recipient. The blood bilirubin level of the infant decreased to normal 2 weeks after operation. No acute rejection occurred. Biliary leakage in the early period after the transplantation was controlled by drainage, and E.coli infection was effectively treated with antibiotics. The donor and recipient are in satisfactory condition to the present., Conclusion: LRLT is advisable for children with biliary atresia.

Objective: To summarize the experience of human orthotopic liver transplantation (OLTx) in treatment of patients with end-stage hepatic disease and their perioperative management., Methods: A retrospective analysis was made on OLTx performed in 77 patients from April 1993 to September 2001 in our department included combined liver-kidney transplantation (6 patients) and living related liver transplantation (2). Among them, 76 were adults and 1 was infant (67 males and 10 females). The donors included 75 non-heart beating donors and 2 living related donors. Veno-venous bypass was used only in 45 cases during the operation. immunosuppressive agents consisted of cyclosporine, cellcept, ALG corticosteroids and FK506., Results: The one-year survival rates of grafts and recipients with benign hepatic disease were 75%. The recipients this year are surviving with good function of grafts., Conclusion: Liver transplantation is the best therapeutic method for a large number of patients with end-stage hepatic disease in China.

Objective: To investigate human cytomegalovirus infection and genetic variations in glycoprotein B (gB) in liver transplant recipients in south-east China., Methods: EDTA-blood samples were obtained from 21 liver transplant recipients. The semi-nested PCR was used to amplify a region of high sequence variability in the gB gene of human cytomegalovirus (HCMV) followed by direct sequence analysis., Results: Out of the 21 liver transplant recipients, 5 were proved HCMV positive 62 to 180 days after transplantation. The nucleotide and encoded amino acid sequences were compared with published sequences of AD169 and Towne laboratory strains. Within the region sequenced, 2 out of 5 strains possessed a peptide configuration similar to that of strain AD169, while another 2 strains displayed a peptide configuration similar to that of strain Towne. One strain had amino acid substitution, which was different from those of both AD169 and Towne in the cleavage site., Conclusion: Our results provide molecular epidemiological data for HCMV strains circulating among transplant recipients in south-east China.

Objective: To explore cytomegalovirus (CMV) infection in recipients of liver transplantation (LT)., Methods: 30 recipients of LT were screened for the appearance of CMV infection by using ELISA to test anti-CMV-Ab from serum samples and using immunohistochemistry method to test CMV antigen expression and nested-PCR to amplify CMV-DNA from blood samples., Results: Four of 243 samples taken from 30 recipients came out positive of anti-CMV IgG and anti-CMV IgM with a positive rate of 100% and 1.6% respectively. 85 samples resulted in CMV antigen expression (35.0%) with the average antigen index being 4.2+/-3.1/5 X 10(4) WBC. Besides, 99 samples were found to be positive by nested-PCR with a positive rate of 40.7%. 61 samples were found to be simultaneously positive in test of CMV antigen and DNA, with a rate of 25.1%., Conclusions: Infection of CMV is common in recipients of LT. Simultaneous screening of anti-CMV-Ab, CMV-Ag and CMV-DNA after liver transplantation is very important for early diagnosis of CMV infection.

We reported preliminary results of rapid propagation, callus induction and regeneration of Trichosanthes Kirilowii, and its protein analysis. Pre-existing meristerms regenerate shoots very rapidly when grown on MS medium containing 0.5 or 1.0 mg/L 6-BA; calli could be induced from leaf sections when put on MS medium containing 4.0 mg/L 6-BA + 0.2 mg/L IAA; shoots regenerated successfully 30 days after calli induction and the differentiation ratio was one shoot out of every four leaf sections; and all shoots gave rise to roots after removing onto MS medium containing 0.1 mg/L NAA and 100% survived when transplanted into soil. Very excitingly, these plants produced small tubers in one month, where satisfactory expression of TCS protein was detected by Western blot analysis.

Glutathione peroxidases have been thought to function in cellular antioxidant defense. However, some recent studies on Gpx1 knockout (-/-) mice have failed to show a role for Gpx1 under conditions of oxidative stress such as hyperbaric oxygen and the exposure of eye lenses to high levels of H2O2. These findings have, unexpectedly, raised the issue of the role of Gpx1, especially under conditions of oxidative stress. Here we demonstrate a role for Gpx1 in protection against oxidative stress by showing that Gpx1 (-/-) mice are highly sensitive to the oxidant paraquat. Lethality was already detected within 24 h in mice exposed to paraquat at 10 mg.kg-1 (approximately (1)/(7) the LD50 of wild-type controls). The effects of paraquat were dose-related. In the 30 mg.kg-1-treated group, 100% of mice died within 5 h, whereas the controls showed no evidence of toxicity. We further demonstrate that paraquat transcriptionally up-regulates Gpx1 in normal cells, reinforcing a role for Gpx1 in protection against paraquat toxicity. Finally, we show that cortical neurons from Gpx1 (-/-) mice are more susceptible to H2O2; 30% of neurons from Gpx1 (-/-) mice were killed when exposed to 65 microM H2O2, whereas the wild-type controls were unaffected. These data establish a function for Gpx1 in protection against some oxidative stressors and in protection of neurons against H2O2. Further, they emphasize the need to elucidate the role of Gpx1 in protection against different oxidative stressors and in different disease states and suggest that Gpx1 (-/-) mice may be valuable for studying the role of H2O2 in neurodegenerative disorders.

Microsurgical procedures have been used to examine the function of selected regions of the epididymis. Vasoepididymostomy in the rabbit and rat confirmed that spermatozoa require some exposure to the epididymal environment for normal development of motility and fertilizing ability. In both of these species surgical redirection of spermatozoa from a region just distal to the initial segment into the vas deferens resulted in a significant reduction in sperm viability, motility and fertility compared with untreated animals. Surgical bypass of the distal cauda epididymidis in the rabbit did not significantly reduce the number of spermatozoa in ejaculates or the total sperm output accumulated during exhaustive ejaculations. The capacity of these bypass animals to store spermatozoa may be explained by the post-surgical formation of convolutions in the normally straight segment of the rabbit vas deferens. Vas autografts in the rabbit and rat were used to bypass selected epididymal segments. In rabbits, vascularized grafts were used as bridges to bypass the distal caput and proximal corpus epididymidis. Despite high rates of graft survival and patency in this species, the motility and viability of ejaculated spermatozoa were similar to those observed following high level vasoepididymostomy. In rats, non-vascularized vas grafts also showed a high survival and patency rate. There was a transitory reduction in sperm viability and motility 2 weeks after surgery which was associated with structural changes, especially in the epithelium, in vas bridge autografts during revascularization. Longer term observations (3 and 6 months) showed recovery of epithelial morphology and by three months after surgery, no difference was observed in sperm motility, sperm viability or fertility between experimental groups and controls. Similar results were obtained when epididymoepididymostomy was used as the surgical procedure to bypass the same regions of the epididymis. The evidence considered in this review clearly shows that the proximity of vasoepididymal bypass to the testis is inversely related to the state of maturity of ejaculated spermatozoa. More surprisingly, perhaps, it reveals that exposure of spermatozoa to the distal caput and proximal corpus regions of the epididymis in the rat is not obligatory for spermatozoa to develop a level of maturation in the remaining regions of the epididymis sufficient to ascend the female tract and fertilize ova. This finding suggests that the epididymis is more flexible in its functions than had been considered previously and that sperm maturation may not be as dependent on specific regional functions of the epididymis as had been previously proposed.

The significance of neoangiogenesis in the metastasis of nasopharyngeal carcinoma (NPC) was investigated to clarify the role neovascularity in the prognosis of NPC and the probability of antiangiogenesis preventing NPC from distant metastasis. A group of 52 patients presenting with metastatic NPC were selected and strictly paired one-to-one, in sex, age, T stage, and N stage, with another 52 patients with non-metastatic NPC, who had survived for a long time after therapy. The tumor tissues of all 104 patients were retrieved for computer-assisted, immunohistochemical analysis of tumor vasculature. Counts of the microvessels and the relative area of all microvessels per image were significantly higher in metastatic NPC than they were in curable, non-metastatic NPC, while the average area of the microvessels and their average perimeter of in metastatic NPC were smaller than in non-metastatic disease. No significant difference in any microvessel parameter was found among the various types of metastasis. The alterations of microvessel parameters were significantly linked to the metastasis of NPC. Evaluation of neovascularity by computer image analysis may be helpful in estimating the prognosis of NPC and in determining the indicators for aggressive multimodal treatments.

It is postulated the pulmonary injury in acute necrotizing pancreatitis could be due to oxygen-derived free radicals released during the attack of acute pancreatitis. To elucidate the relationship between oxygen-derived free-radicals and the degree of pulmonary injury in acute necrotizing pancreatitis, lipid peroxide (LPO) in lung tissues, bronchoalveolar lavage fluid and serum were measured in 10 adult Mongrel dogs with acute necrotizing pancreatitis induced by injection of 5% sodium taurocholate into the pancreatic duct. Compared with the controls (n = 10), the weight of lung was increased (241.2 +/- 33.7 g vs 121.2 +/- 51.4 g, P < 0.05). LPO of bronchoalveolar lavage fluid (BALF) was higher (3.56 +/- 1.64 mumol/L vs 0.75 +/- 0.31 mumol/L, P < 0.05), LPO in peripheral and portal vein 10 hours after induction of acute pancreatitis was also significantly increased. The LPO content of lung tissue was not higher than that of the control group, but hemorrhage within alveolar space and infiltration by polymorphonuclear and phagocytic cells was seen on histological examination. In conclusion, lipid peroxidation could be an important cause of pulmonary injury in acute necrotising pancreatitis.

We conducted a prospective study to validate our previous finding that serum urea and plasma glucose levels on admission could predict the outcome of acute pancreatitis. Forty-two (24%) of 176 patients developed complications related to the attack of acute pancreatitis and were classified as having severe disease. By logistic regression analysis of 17 admission parameters, serum urea and plasma glucose levels were again the factors with independent significance in defining the outcome. By adopting the same cutoff levels as in our previous study (serum urea level greater than 7.4 mmol/L and plasma glucose level greater than 11.0 mmol/L), and the presence of either factor above the cutoff level as indicative of severe disease, the sensitivity of prediction was 79%, specificity 67%, and overall accuracy 70%. All the deaths were correctly predicted by this urea/glucose criteria. The overall accuracy was also found to be comparable with those of the APACHE II (cutoff level greater than 11) and Ranson's scoring systems. We conclude that the simple prognostic criteria for acute pancreatitis were validated; these criteria have the potential to stratify risk rapidly at the time of admission for patients who might benefit from an aggressive interventional protocol.

Hepaticocutaneous jejunostomy offers the advantage of permanent percutaneous access to the biliary tract in patients with complex biliary problems. The long-term value, however, has not been assessed. In 41 patients who underwent this procedure for intrahepatic stones, there was no hospital mortality, and the postoperative morbidity rate was 10%. The presence of the cutaneous stoma facilitated postoperative flexible choledochoscopy for dilatation of biliary strictures and extraction of residual stones. On follow-up, symptoms recurred in 12 patients (29%) at a median time of 27 months (range: 1 to 97 months). Reconstruction of the cutaneous stoma and flexible choledochoscopy via the jejunal loop helped to resolve the acute cholangitis (n = 4) and to eradicate recurrent stones in all cases. Hepaticocutaneous jejunostomy was also beneficial in the management of three patients who experienced further recurrence of symptoms. There was no major difficulty in reconstructing the cutaneous stoma and in performing choledochoscopy via the jejunal loop. The overall complication rate related to the cutaneous stoma was 15%. Repeat laparotomy for recurrent disease was required in only one patient who underwent a second bilio-enteric anastomosis for a nondilatable left duct stricture. We conclude that hepaticocutaneous jejunostomy is a valuable procedure in the management of hepatolithiasis.

Background: Most patients with acute biliary pancreatitis have stones in the biliary tract or ampulla of Vater. Because these stones may be passed spontaneously soon after a patient is admitted to the hospital, the importance of early operative removal is not known. We tested the hypothesis that endoscopic papillotomy within 24 hours of admission decreased the incidence of complications in patients with acute biliary pancreatitis., Methods: We studied 195 patients with acute pancreatitis who were randomly assigned to one of two groups: 97 patients underwent within 24 hours after admission emergency endoscopic retrograde cholangiopancreatography (ERCP) followed by endoscopic papillotomy for ampullary and common-bile-duct stones, and 98 patients received initial conservative treatment and selective ERCP with or without endoscopic papillotomy only if their condition deteriorated., Results: One hundred twenty-seven patients ultimately proved to have biliary stones. Emergency ERCP with or without endoscopic papillotomy resulted in a reduction in biliary sepsis as compared with conservative treatment (0 of 97 patients vs. 12 of 98 patients, P = 0.001). The decrease in biliary sepsis occurred both in patients predicted to have mild pancreatitis (0 of 56 patients in the group that received emergency ERCP vs. 4 of 58 patients in the conservative-treatment group, P = 0.14) and in patients predicted to have severe pancreatitis (0 of 41 patients vs. 8 of 40 patients, P = 0.008). In all patients who had unrelenting biliary sepsis, persistent ampullary or common-bile-duct stones were identified. There were no major differences in the incidence of local complications (10 patients in the group that received emergency ERCP vs. 12 patients in the conservative-treatment group) or systemic complications (10 patients vs. 14 patients) of acute pancreatitis between the two groups, but the hospital mortality rate was slightly lower in the group undergoing emergency ERCP with or without endoscopic papillotomy (5 patients vs. 9 patients, P = 0.4)., Conclusions: Emergency ERCP with or without endoscopic papillotomy is indicated in the treatment of patients with acute pancreatitis.

Skin tests including immediate patch test (IPT), skin prick test (SPT), or intradermal test (IT) with penicillin G(PenG) and SPT with benzylpenicilloyl human serum albumin (BPO) were done in 54 patients with history of anaphylactic reaction to penicillin or shock of unknown cause. 26 patients with penicillin allergy were diagnosed. BPO specific IgE measured with ELISA gave a lower positive rate in detecting penicillin allergy as compared with the tests mentioned above. The results of skin tests in 26 patients showed that IPT with 500 IU/ml of PenG was not only accurate but also safe. Because no skin injury occurred and PenG residue could be washed out, the amount of PenG penetrated into skin is very small, thus, adverse reactions were very few. It is recommended that IPT with PenG in 500 IU/ml concentration is performed at the beginning of skin tests. If negative, SPT and then IT both with a solution of 500 IU/ml concentration are carried out, until a positive reaction occurs. This procedure is relatively accurate, simple and safe.

Allergenicity of three plant abortifacient proteins, trichosanthin (TCS), alpha-momorcharin (alpha-MMC) and beta-momorcharin (beta-MMC) and the kinetics of IgE antibody response against these proteins were studied in C57BL/6N and BALB/cAn mice. These proteins were purified from Chinese medicinal herbs, characterized by biochemical and immunological procedures and found to be homogeneous by PAGE, SDS-PAGE and immunoelectrophoresis. The results obtained were summarized as follows: 1) different levels of IgE antibody responses against TCS, alpha-MMC and beta-MMC were induced when they were injected i.p. as antigen adsorbed onto AI(OH)3 gel into BALB/cAn or C57BL/6N mice on day 0 and day 28; 2) the allergenicity of the proteins was in the order of beta-MMC greater than alpha-MMC greater than TCS; 3) two injections of 10 micrograms or 50 micrograms of TCS or alpha-MMC given at 4 weeks interval without adjuvant elicited a low or undetectable PCA titer, while the same dose given weekly for 5 weeks resulted in high levels of IgE production; and 4) on the basis of PCA observations, no cross immunological reactivity among these three proteins was found, it seemed to support that TCS, alpha-MMC and beta-MMC may be alternately used for inducing abortion.

Experimental study had been conducted on 18 healthy adult mongral dogs of both sex, weighting from 7.5-11 kg, randomly divided into four groups. Experimental models of acute hemorrhagic necrotizing pancreatitis (AHNP) were established by retrograde injecting 1 ml/kg of sodium taurocholate directly into the pancreatic duct. The dogs were treated respectively with intravenous infusion of Salviae miltiorrhizae (5 g/kg), 654-2(5 mg/kg) or normal saline. The results showed that PaO2, PaCO2 and pH did not change in early stage of AHNP. The contents of lactic acid dehydrogenase (LDH), albumin and lipid peroxide (LPO) of bronchoalveolar lavage fluid in the AHNP group were significantly higher than that of Salviae miltiorrhizae group (P less than 0.05). The necrosis and disruption of conjunction of endothelial cells resulting from the defects of vascular wall were noted under transmission electron microscope. Both pulmonary vascular and type II pneumocyte were normal in the Salviae miltiorrhizae group. These results suggested that Salviae miltiorrhizae possess the effect of protecting endothelial cells of pulmonary vascular and type II pneumocyte, which could function as scavenger of oxygen-derived free radicals.