AIM: To investigate the expression of toll-like receptor 4(TLR4) and MD-2 gene and protein in Kupffer cells (KCs)and their role in ischemia-reperfusion (IR) injury of ratliver graft.METHODS: KCs were isolated at 0 (co...AIM: To investigate the expression of toll-like receptor 4(TLR4) and MD-2 gene and protein in Kupffer cells (KCs)and their role in ischemia-reperfusion (IR) injury of ratliver graft.METHODS: KCs were isolated at 0 (control group), 2, 12,24 h (IR group) following IR in rat liver graft, mRNA expressionof TLR4 and MD-2 was detected by RT-PCR analysis, proteinexpression of TLR4/MD-2 was detected by flow cytometric(FCM) analysis, and tumor necrosis factor-(~ (TNF-(~) levelin supernatant was measured by ELISA. Then isolated KCswere incubated with anti-TLR4 polyclonal antibody (anti-TLR4 group), and TNF-(~ level was measured again.RESULTS: The mRNA and protein expression of TLR4/MD-2 and the level of TNF-(~ in IR group increased significantlyat 2 h following IR, and reached the maximum at 12 h, andslightly decreased at 24 h, but were still significantly higherthan those in the control group (P<O.01). The expression ofthese factors was markedly decreased after anti-TLR4antibody treatment as compared with the IR group (P<O.01).CONCLUSION: Lipopolysaccharide (LPS) following IR canup-regulate TLR4/MD-2 gene and protein expression inKCs, and synthesize cytokine TNF-(~. Anti TLR4 antibodycan inhibit the production of TNF-(~ induced by LPS. TLR4and its partner molecule MD-2 may play an important rolein Kupffer cell activation and IR injury.展开更多
AIM:The diagnosis of cholangiocarcinoma is often difficult,making management approaches problematic. A reliable serum marker for cholangiocarcinoma would be a useful diagnostic test. The aims of our study were to eval...AIM:The diagnosis of cholangiocarcinoma is often difficult,making management approaches problematic. A reliable serum marker for cholangiocarcinoma would be a useful diagnostic test. The aims of our study were to evaluate the usefulness of a serum CA19-9 determination in the diagnosis of cholangiocarcinoma.METHODS: We prospectively measured serum CA19-9 and CEA concentrations in patients with cholangiocarcinoma (n=35), benign biliary diseases (n=92), and healthy individuals (n=15). Serum CA19-9 and CEA concentrations were measured by an immunoradiometric assay without knowledge of the clinical diagnosis.RESULTS:The sensitivity of a CA19-9 value>37KU·L^-1 and a CEA value >22μg·L^-1 in diagnosing cholangiocarcinoma were 77.14% and 68.57%, respectively. When compared with the benign biliary diseases group,the true negative rates of serum CA19-9 and CEA were 84.78% and 81.52%,respectively. The false positive rates of serum CA19-9 and CEA were 15.22% and 18.48%, whereas the accuracy of serum CA19-9 and CEA were 82.68% and 77.95%,respectively. Serum CA19-9 and CEA concentrations were significantly elevated (P<0.001 and P<0.05) in patients with cholangiocarcinoma (290.31±5.34KU·L^-1 and 36.46±18.03μg·L^-1) compared with patients with benign biliary diseases (13.38±2.59KU·L^-1 and 13.84±3.85μg·L^-1) and healthy individuals (12.78±3.69KU·L^-1 and 11.48±3.37μg·L^-1). In 15 patients undergoing curative resection of cholangiocarcinoma,the mean serum CA19-9 concentration was decreased from a preoperative level of 286.41±4.36KU·L^-1 to a postoperative level of 62.01±17.43KU·L^-1 (P<0.001), and the mean serum CEA concentration from 39.41±24.35μg·L^-1 to 28.69±11.03μg·L6-1(P<0.05). In patients with cholangiocarcinoma,however, no correlation was found between serum CEA and CA19-9 concentrations (r=-0.036).CONCLUSION:These data suggest that the serum CA19-9 determination is a useful addition to the available tests for the differential diagnosis of cholangiocarcinoma. Serum CA19-9 is an effective tumor marker in diagnosing cholangiocarcinoma,deciding whether the tumor has been radically resected and monitoring effect of treatment.展开更多
AIM: To study the anti-tumor effect of resveratrol and in combination with 5-FU on murine liver cancer. METHODS: Transplantable murine hepatoma22 model was used to evaluate the anti-tumor activity of resveratrol (RES)...AIM: To study the anti-tumor effect of resveratrol and in combination with 5-FU on murine liver cancer. METHODS: Transplantable murine hepatoma22 model was used to evaluate the anti-tumor activity of resveratrol (RES) alone or in combination with 5-FU in vivo. H22 cell cycles were analyzed with flow cytometry. RESULTS: Resveratrol could inhibit the growth of murine hepatoma22, after the mice bearing H22 tumor were treated with 10 mg/kg or 15 mg/kg resveratrol for ten days, and the inhibition rates were 36.3% (n = 10) and 49.3% (n = 9), respectively, which increased obviously compared with that in control group (85+22 vs 68+17, P<0.01). RES could induce the S phase arrest of H22 cells, and increase the persentage of cells in S phase from 59.1% (n = 9) to 73.5% (n = 9) in a dose-dependent manner (P<0.05). The enhanced inhibition of tumor growth by 5-FU was also observed in hepatoma22 bearing mice when 5-FU was administered in combination with 10 mg/kg resveratrol. The inhibition ratesfor 20 mg/kg or 10 mg/kg 5-FU in combination with 10 mg/kg resveratrol were 77.4% and 72.4%, respectively, compared with the group of 20 mg/kg or 10 mg/kg 5-FU alone, in which the inhibition rates were 53.4% and 43.8%, respectively (n = 8). There was a statistical significance between the combination group and 5-FU group. CONCLUSION: RES could induce the S phase arrest of H22 cells and enhance the anti-tumor effect of 5-FU on murine hepatoma22 and antagonize its toxicity markedly. These results suggest that resveratrol, as a biochemical modulator to enhance the therapeutic effects of 5-FU, may be potentially useful in cancer chemotherapy.展开更多
AIM: To observe the effect of resveratrol on nuclear factor Kappa-B (NF-κB) activation and the inflammatory response in sodium taurocholate-induced pancreatitis in rats. METHODS: Seventy-two male SD rats were randoml...AIM: To observe the effect of resveratrol on nuclear factor Kappa-B (NF-κB) activation and the inflammatory response in sodium taurocholate-induced pancreatitis in rats. METHODS: Seventy-two male SD rats were randomly divided into three groups: sham operation group (control), severe acute pancreatitis (SAP) group, and severe acute pancreatitis group treated with resveratrol (RES). A SAP model was established by injecting 4% sodium taurocholate 1 mL/kg through puncturing the pancreatic duct. In Res group, Res was given at 30 mg/kg b.m. intraperitoneally after the SAP model was successfully established. Eight animals from each group were sacrificed at 3, 6 and 12 h after modeling. The expression of NF-κB activation of pancreas was detected by irnmunohistochemical staining, whereas the levels of TNF-α and IL-8 in pancreatic tissues were estimated by radioimrnunoassay. The pathological changes of pancreas and lungs were examined microscopically. RESULTS: Much less hyperemia, edema, dust-colored necrotic focus and soaps were noticed in pancreas in RES group than in SAP group. In RES group, hemorrhage, exudates and infiltration of inflammatory cells in pancreas and interstitial edema, destruction of alveolar wall in lung were significantly less than in SAP group. In the SAP group, the activation of NF-κB in pancreatic tissues was enhanced significantly at any measure point compared with control group (64.23±10.72% vs 2.56±0.65%, 55.86±11.34% vs 2.32±0.42%, 36.23±2.30% vs 2.40±0.36%,P<0.01), TNF-α, IL-8 were also increased and reached their peak at 6 h and then declined. The activation of NF-κB and the levels of TNF-α and IL-8 in RES group were significantly lower than those in SAP group (P<0.01): activation (52.63±9.45% vs 64.23±10.72%, 40.52±8.40% vs 55.86±11.34%, 29.83±5.37% vs 36.23±2.30%), TNF-α (132.76±15.68 pg/mL vs 158.36±12.58 pg/mL, 220.32±23.57 pg/mL vs 247.67± 11.62 pg/mL, 175.68±18.43 pg/mL vs 197.35±12.57 pg/mL) and IL-8 (0.62±0.21 μg/L vs 0.83±0.10 μg/L, 1.10±0.124 μg/L vs1.32±0.18 μg/L, 0.98±0.16 μg/L vs 1.27±0.23μg/L). CONCLUSION: The activation of NF-KB is involved in the inflammatory response of rats with SAP. Resveratrol could effectively inhibit the expression of NF-κB activation, alleviate the severity of SAP through its anti-inflammatory effects and regulate the inflammatory mediators.展开更多
AIM: To study the protective effects of tumor necrosis factor α (TNFα) antibody and ulinastatin on liver ischemic reperfusion in rats.METHODS: One hundred and twenty male SD rats were randomly divided into four grou...AIM: To study the protective effects of tumor necrosis factor α (TNFα) antibody and ulinastatin on liver ischemic reperfusion in rats.METHODS: One hundred and twenty male SD rats were randomly divided into four groups: normal control group, ischemic group, TNFα antibody group and TNFα antibody + ulinastatin group. The animals were killed at 0, 3, 6, 9, 12 h after ischemia for 60 min and followed by reperfusion. Serum alanine aminotransferase (ALT), malondialdehyde (MDA) and liver histopathology were observed.RESULTS: After ischemic reperfusion, the serum ALT and MDA were remarkably increased, and the hepatic congestion was obvious. Treatment of TNFα antibody and ulinastatin could significantly decrease serum ALT and MDA levels, and relieve hepatic congestion.CONCLUSION: Ulinastatin and TNFα antibody can suppress the inflammatory reaction induced by hepatic ischemic reperfusion, and have protective effects on rat hepatic ischemic reperfusion injury.展开更多
AIM: To investigate the protective effect of ischemic preconditioning (IPC) on hepatocellular carcinoma (HCC) patients with cirrhosis undergoing hepatic resection under hepatic inflow occlusion (HIO) and its possible ...AIM: To investigate the protective effect of ischemic preconditioning (IPC) on hepatocellular carcinoma (HCC) patients with cirrhosis undergoing hepatic resection under hepatic inflow occlusion (HIO) and its possible mechanism.METHODS: Twenty-nine consecutive patients with resectable OHCC were randomized into two groups: IPC group: before HIO, IPC with 5 min of ischemia and 5 min of reperfusion was given; control group: no IPC was given. Uver functions,hepatic Caspase-3 activity, and apoptotic cells were compared between these two groups.RESULTS: On postoperative days (POD) 1, 3 and 7, the aspartate transaminase (AST) and alanine transaminase (ALT) levels in the IPC group were significantly lower than those in the control group (P<0.05). On POD 3 and 7, the total bilirubin level in the IPC group was significantly lower than that in the control group (P<0.05). On POD 1, the albumin level in the IPC group was higher than that in the control group (P = 0.053). After 1 h of reperfusion, both hepatic Caspase-3 activity and apoptotic sinusoidal endothelial cells in the IPC group were significantly lower than those in the control group (P<0.05).CONCLUSION: IPC has a potential protective effect on HCC patients with cirrhosis. Its protective mechanism underlying the suppression of sinusoidal endothelial cell apoptosis is achieved by inhibiting Caspase-3 activity.展开更多
AIM: To study changes in characteristics of colorectal carcinoma during the metastatic process and to investigate the correlation between cell proliferation activity and metastatic ability of patients with Dukes′ sta...AIM: To study changes in characteristics of colorectal carcinoma during the metastatic process and to investigate the correlation between cell proliferation activity and metastatic ability of patients with Dukes′ stage C or D.METHODS: Formalin fixed and paraffin embedded materials of primary tumors and corresponding lymph node metastases resected from 56 patients with Dukes′ stage C or D of colorectal carcinoma were stained immunohistochemically with proliferating cell nuclear antigen (PCNA) and CD44variant exon 6 (CD44v6).RESULTS: Thirty-one of 56 patients (55.4 %) expressed PCNA in the primary sites and 36 of 56 patients (64.3 %)expressed PCNA in the metastatic lymph nodes. A significant relation in PCNA expression was observed between the primary site and the metastatic lymph node (0.010＜P＜0.025).Forty-one of 56 patients (73.2 %) expressed CD44v6 in the primary site and 39 of 56 patients (69.6 %) expressed CD44v6 in the metastatic lymph node. There was also an significant relationship of CD44v6 between the primary site and the metastatic lymph node (0.005＜P＜0.010). No difference was observed between expression of CD44v6 and PCNA in the primary site (0.250＜P＜0.500).CONCLUSION: This study partially demonstrates that tumor cells in metastatic lymph node of colorectal carcinoma still possess cell proliferation activity and metastatic ability of tumor cells in primary site. There may be no association between cell proliferation activity and metastatic ability in colorectal carcinoma.展开更多
AIM: To evaluate the effects of nitroester drugs on human sphincter of Oddi (SO) motility and their antagonistic effects against morphine which shows excitatory effect on Oddi's sphincter motility.METHODS: The eff...AIM: To evaluate the effects of nitroester drugs on human sphincter of Oddi (SO) motility and their antagonistic effects against morphine which shows excitatory effect on Oddi's sphincter motility.METHODS: The effects of these drugs on SO were evaluated by means of choledochofiberoscopy manometry.A total of 67 patients having T-tubes after cholecystectomy and choledochotomy were involved in the study, they were randomly divided into glyceryl trinitrate (GTN) group,isosorbide dinitrate (ISDN) group, pentaerythritol tetranitrate (PTN) group, morphine associated with GTN group, morphine associated with ISDN group and morphine associated with PTN group. Basal pressure of Oddi's sphincter (BPOS), amplitude of phasic contractions (SOCA), frequency of phasic contractions (SOF), duration of phasic contractions (SOD), duodenal pressure (DP) and common bile duct pressure (CBDP) were scored and analyzed. Morphine was given intramuscularly while nitroester drugs were applied sublingually.RESULTS: BPOS and SOCA decreased significantly after administration of ISDN and GTN, BPOS reduced from 10.95±7.49 mmHg to 5.92±4.04 mmHg (P＜0.05) evidently after application of PTN. BPOS increased from 7.37±5.58mmHg to 16.60±13.87 mmHg, SOCA increased from 54.09±38.37 mmHg to 100.70±43.51 mmHg, SOF increased from 7.15±3.20 mmHg to 10.38±2.93 mmHg and CBDP increased 3.75±1.95 mmHg to 10.49±8.21 mmHg (P＜0.01)evidently after injection of morphine. After associated application of ISDN and GTN, the four indications above decreased obviously. As for application associated with PTN,SOCA and SOF decreased separately from 100.64±44.99mmHg to 66.17±35.88 mmHg and from 10.70±2.76 mmHg to 9.04±1.71 mmHg (P＜0.05) markedly.CONCLUSION: The regular dose of GTN, ISDN and PTN showed inhibitory effect on SO motility, morphine showed excitatory effect on SO while GTN, ISDN and PTN could antagonize the effect of morphine. Among the three nitroester drugs, the effect of ISDN on SO was most significant.展开更多
AIM:Intrahepatic extension is the main cause of liver failure and death in hepatocellular carcinoma patients.The small GTPase Rho and one of its effector molecules ROCK regulate cytoskeleton and actomyosin contractili...AIM:Intrahepatic extension is the main cause of liver failure and death in hepatocellular carcinoma patients.The small GTPase Rho and one of its effector molecules ROCK regulate cytoskeleton and actomyosin contractility,and play a crucial role in cell adhesion and motility.We investigated the role of small GTPase Rho in biological behaviors of hepatocellular carcinoma to demonstrate the importance of Rho in cancer invasion and metastasis.METHODS:Using Western blotting,we quantitated Rho protein expression in SMMC-7721 cells induced by Lysophosphatidic acid (LPA).Furthermore,we examined the role of Rho signaling in regulating the motile and invasive properties of tumor cells.RESULTS:Rho protein expression was stimulated by LPA.Using the Rhotekin binding assay to assess Rho activation,we observed that the level of GTP-bound Rho was elevated transiently after the addition of LPA,and Y-27632 decreased the level of active Rho.LPA enhanced the motility of tumorcells and facilitated their invasion.Rho played an essential role in the migratory process,as evidenced by the inhibition of migration and motility of cancer cells by a specific inhibitor of ROCK,Y-27632.CONCLUSION: The finding that invasiveness of hepatocellular carcinoma is facilitated by the Rho/Rho-kinase pathway is likely to be relevant to tumor progression and Y-27632 may be a new potential effective agent for the prevention of intrahepatic extension of human liver cancer.展开更多
AIM: To assess the effects of intramuscular analgesics(morphine, Ap-237, pethidine and tramadol) on humanOddi's sphincter motility with choledochoscope manometry.METHODS: A total of 70 patients having T tubes afte...AIM: To assess the effects of intramuscular analgesics(morphine, Ap-237, pethidine and tramadol) on humanOddi's sphincter motility with choledochoscope manometry.METHODS: A total of 70 patients having T tubes aftercholecystectomy and choledochotomy were assessed bycholedochoscope manometry. They were randomly dividedinto morphine group, Ap-237 group, pethidine group andtramadol group. Basal pressure of Oddi's sphincter (BPOS),amplitude of phasic contractions (SOCA), frequency ofphasic contractions (SOF), duration of phasic contractions(SOD), duodenal pressure (DP) and common bile ductpressure (CBDP) were scored and analyzed. All narcoticanalgesic drugs were administered intramuscularly.RESULTS: Levels of BPOS, SOCA and SOF were increasedafter injection of morphine and Ap-237 (P<0.05), level of CBDPwas increased from 4.97+3.87 mmHg to 8.62+7.43 mmHg(10 min later) and 7.32+5.95 mmHg (20 min later) afterinjection of morphine (P<0.01). No apparent changeoccurred after intramuscular injection of pethidine. Level ofBPOS was increased from 7.01+5.50 mmHg to 2.87+2.78 mmHg10 min after injection of tramadol and SOCA was decreasedfrom 63.34+35.29 mmHg to 45.90+27.86 mmHg (10 minlater, P<0.05) and 35.97+24.30 (20 min later, P<0.01) afteradministration of tramadol.CONCLUSION: All these findings indicate that Oddi'ssphincter manometry via choledochoscope is a practicaland new way to study the dynamics of Oddi' s sphincter.The regular dose of morphine and Ap-237 could increaseBPOS, SOF and SOCA. Morphine could increase the levelof CBDP, demonstrating an excitatory effect on the sphincterof Oddi. Pethidine had no effect on Oddi's sphincter motility.Tramadol shows an inhibitory effect on the motility of thesphincter of Oddi and decreases levels of BPOS and SOCA.展开更多
AIM:To study the effect of matrine on activation of Kupffer cell during cold ischemia and reperfusion injury in rat orthotopic liver transplantation(OLT).
AIM: To investigate the mechanism of resveratrol underlying the microcirculation disorder and lung injury following severe acute pancreatitis (SAP). METHODS: Twenty-four rats were divided into 3 groups (SAP, sham and ...AIM: To investigate the mechanism of resveratrol underlying the microcirculation disorder and lung injury following severe acute pancreatitis (SAP). METHODS: Twenty-four rats were divided into 3 groups (SAP, sham and resveratrol groups) randomly. SAP model was established by injecting 4% sodium taurocholate l mL/kg through puncturing pancreatic ducts. Sham (control) group (8 rats) was established by turning over the duodenum. Resveratrol was given at 0.1 mg/kg b.m. intraperitoneally. Rats were sacrificed 9 h after SAP was induced. Blood samples were obtained for hemorrheological examination. Lung tissues were used for pathological observation, and examination of microvascular permeability, dry/wet ratio and myeloperoxidase (MPO) activity. Gene expression of intercellular adhesion molecule-1 (ICAM-1) was detected by RT-PCR. RESULTS: Compared with SAP group, resveratrol relieved the edema and infiltration of leukocytes in the lungs. Resveratrol improved markers of hemorrheology: high VTB (5.77±1.18 mPas vs9.49±1.34 mPas), low VTB (16.12±3.20 mPas vs30.91±7.28 mPas), PV (4.69±1.68 mPas vs 8.00±1.34 mPas), BSR (1.25±0.42 mm/h vs50.03±0.03 mm/h), VPC (54.67±3.08% vs 62.17±3.39%), fibrinogen (203.2?7.8 g/ L vs 51.3±19.1 g/L), original hemolysis (0.45±0.02 vs 0.49±0.02), and complete hemolysis (0.41±0.02 vs 0.43±0.02) (P<0.05). Resveratrol decreased the OD ratio of ICAM-1 gene (0.800±0.03 vs 1.188±0.10), dry/wet ratio (0.74±0.02 vs 0.77±0.03), microvascular permeability (0.079±0.006 vs 0.112±0.004) and MPO activity (4.42±0.32 vs 5.03±0.51) significantly (P<0.05). CONCLUSION: Resveratrol can improve the microcirculation disorder of the lung by decreasing leukocyte-endothelial interaction, reducing blood viscosity, improving the decrease of blood flow, and stabilizing erythrocytes in SAP rats. It may be a potential candidate to treat SAP and its severe complications (ALI).展开更多
AIM: To sum up the clinical characteristics of patients with central pontine myelinolysis (CPM) after orthotopic liver transplantation (OLT) and to document the possible causes of CPM.METHODS: Data of 142 patients und...AIM: To sum up the clinical characteristics of patients with central pontine myelinolysis (CPM) after orthotopic liver transplantation (OLT) and to document the possible causes of CPM.METHODS: Data of 142 patients undergoing OLT between January 1999 to May 2003 were analyzed retrospectively.Following risk factors during perioperation were analyzed in patients with and without CPM: primary liver disease,preoperative serum sodium level, magnesium level and plasma osrnolality, fluctuation degree of serum sodium concentration, and irnmunosuppressive drug level, etc.RESULTS: A total of 13 (9.2%) neurologic symptoms appeared in 142 patients post-operation including 5 cases (3.5%) with CPM and 8 cases (5.6%) with cerebral hemorrhage or infarct. Two patients developing CPM after OLT had a hyponatrernia history before operation (serum sodiurn<130 mmol/L), their mean serum sodium level was 130.6±5.54 mmol/L. The serum sodium level was significantly lower in CPM patients than in patients without neurologic complications or with cerebral hemorrhage/infarct (P<0.05).The increase in serum sodiurnduring perioperative 48 h after OLT in patients wibh CPM was significantly greater than bhat in patients with cerebral hemorrhage/infarct but without neurologic complications (19.5+6.54 retool/L, 10.1+6.43 retool/L,4.5±4.34 mmol/L, respectively, P<0.05). Plasma osrnolality was greatly increased postoperation in patients with CPM.Hypomagnesernia was noted in all patients perioperation,but there were no significant differences between groups.The duration of operation on patients with CPM was longer than that on others (492+190.05 rnin, P<0.05). Cyclosporin A (CsA) levels were normal in all patients, but there were significant differences between patients with or without neurologic complications (P<0.05).CONCLUSION: CPM may be more prevalent following liver transplantation. Although the diagnosis of CPM after OLT can be made by overall neurologic evaluations including magnetic resonance imaging (MRI) of the head, the mortality is still very high. The occurance of CPM may be associated with such factors as hyponatremia, rapid rise of serum sodium concentration, plasma osmolality increase postoperation, the duration of operation, and high CsA levels.展开更多
AIM: To study the mechanism and effect of nuclear factor-κB (NF-κB) activation and inflammatory response on the extended cold-preserved graft injury after orthotopic liver transplantation (OLT). METHODS: OLT was per...AIM: To study the mechanism and effect of nuclear factor-κB (NF-κB) activation and inflammatory response on the extended cold-preserved graft injury after orthotopic liver transplantation (OLT). METHODS: OLT was performed in rats with varying time of cold ischemia grafts (6, 18 and 24 h in University Wisconsin solution at 4 ℃). We determined the time of NF-κB activation and expression of tumor necrosis factor-α (TNF-α), cytokineinducible neutrophil chemoattractant (CINC), and intercellular adhesion molecule-1 (ICAM-1) within 6 h after reperfusion. Serum alarming aminotransferase (ALT), neutrophil sequestration, circulating neutrophil CD11b and L- selectin expression were also evaluated. RESULTS: The accumulation of neutrophils in the graft was significantly increased in the 18 h and 24 h cold-ischemia groups within 0.5 h after reperfusion, compared with the 6 h group. But the strongly activated neutrophils was slightly increased at 2 h after reperfusion and remained at high levels 4 h after reperfusion, which was synchronized with the common situation of recipients after transplantation. Prolonged cold-preservation did not affect neutrophil accumulation and activation. NF-κB activation preceded the expression of TNF-α, CINC, and ICAM-1 in the liver, which was significantly increased with prolonged cold preservation. In prolonged cold preserved grafts, prominently elevated NF-κB activation occurred at 0.5 h and 1 h, compared with that at 2 h after reperfusion, which was consistent with greatly increased intrahepatic TNF-α response. CONCLUSION: NF-κB activation is correlated with the expression of TNF-α, CINC, and ICAM-1 in vivo in OLT rats. Extended cold preservation of grafts might up-regulate TNF-α, CINC, and ICAM-1 expression in the grafts, most probably through elevated NF-κB activation, and might contribute to neutrophil infiltration in the grafts after reperfusion. Elevated NF-κB activity is harmful to inflammatory response in the grafts, and inhibited NF-κB activity might protect against early graft injury after liver transplantation.展开更多
AIM: To develop an in vitro three-dimensional (3-D) angiogenesis system to analyse the capillary sprouts induced in response to the concentration ranges of basic fibroblast growth factor (bFGF) and vascular endothelia...AIM: To develop an in vitro three-dimensional (3-D) angiogenesis system to analyse the capillary sprouts induced in response to the concentration ranges of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) and to quantify their synergistic activity.METHODS: Microcarriers (MCs) coated with human microvascular endothelial cells (HMVECs) were embedded in fibrin gel and cultured in 24-well plates with assay media. The growth factors bFGF, or VEGF, or both were added to the system. The wells (n = 8/group) were digitally photographed and the average length of capillary-like sprouts (ALS) from each microcarrier was quantitated.RESULTS: In aprotinin-stabilized fibrin matrix, human microvascular endothelial cells on the MCs invaded fibrin,forming sprouts and capillary networks with lumina. The angiogenic effects of bFGF or VEGF were dose-clependent in bhe range from 10 to 40 ng/mL. At d 1, 10 ng/mL of bFGF and VEGF induced angiogenesis with an ALS of 32.13±16.6 μm and 43.75±27.92 μm, respectively, which were significantly higher than that of the control (5.88±4.45 μm, P<0.01),and the differences became more significant as the time increased. In addition, the combination of 10 ng/mL of bFGF and VEGF each induced a more significant effect than the summed effects of bFGF (10 ng/mL) alone and VEGF (10 ng/mL) alone when analyzed using SPSS system for general linear model (GLM) (P= 0.011), and bhat also exceeded the effects by 20 ng/mL of either bFGF or VEGF.CONCLUSION: A microcarrier-based in vitro threedimensional angiogenesis model can be developed in fibrin.It offers a unique system for quantitative analysis of angiogenesis. Both bFGF and VEGF exert their angiogenic effects on HMVECs synergistically and in a dose-dependent manner.展开更多
AIM:To explore the relationship between angiogenesis and biological behaviors of primary gallbladder carcinoma (PGBC),the relationship between the expression of inducible nitric oxide synthase (iNOS) and biological be...AIM:To explore the relationship between angiogenesis and biological behaviors of primary gallbladder carcinoma (PGBC),the relationship between the expression of inducible nitric oxide synthase (iNOS) and biological behaviors of PGBC and its relationship with the expression of iNOS and angiogenesis of PGBC.METHODS: The expression of iNOS and micro-vessel density (MVD) were assessed by immunohistochemical method and image analysis system in 40 specimens of PGBC and in 8 specimens of normal gallbladder. The immunostaining results and related clinicopathologic materials were analyzed by statistical methods.RESULTS: MVD in PGBC was significantly higher than that in normal gallbladder tissue (46±14 vs 14±6, P<0.05), and was not related with age, gender, tumor size and histological type. MVD of poorly and undifferentiated tumor tissues was higher than that of moderately-differentiated and well-differentiated tumor tissues (52±9 vs43±9 vs33±6, P<0.01).MVD of Nevin IV and V stages was higher than that of NevinI, Ⅱ and Ⅲ stages (52±8 vs 37±13, P<0.01). MVD of cases with lymphatic or liver metastasis was significantly higher than that without liver metastasis (55±6 vs 42±10, P<0.05) or lymphatic metastasis (53±8 vs38±8, P<0.01). The positive level index (PLI) of iNOS in PGBC was 0.435±0.134, and was not related with age, gender, tumor size, histological type,differentiation and clinical stage of PGBC. The PLI of iNOS in cases with lymphatic metastasis was higher than that without lymphatic metastasis (0.573±0.078 vs 0.367±0.064,P<0.01). The PLI of iNOS in cases with liver metastasis was higher than that without liver metastasis (0.533±0.067 vs0.424±0.084, P<0.05). There was a significant correlation between PLI of iNOS and MVD in PGBC (P<0.05).CONCLUSION:Angiogenesis of PGBC is significantly related to the biological behaviors of PGBC. The expression of iNOS is related to the biological behaviors of PGBC. The detection of MVD and the expression of iNOS in PGBC can be used as parameters to determine the degree of malignancy and prognosis.展开更多
AIM: To study oval cells' pathological characteristics and relationship with the occurrence of hepatocellular carcinoma (HCC); to observe the form and structural characteristics of oval cells; to explore the expre...AIM: To study oval cells' pathological characteristics and relationship with the occurrence of hepatocellular carcinoma (HCC); to observe the form and structural characteristics of oval cells; to explore the expression characteristics of C-kit, PCNA mRNA and c-mycgene during the occurrence and development of HCC and the effect of ulinastatin (UTI) on C-kit and PCNA expression.METHODS: One hundred and twenty-five SD rats fed on 3,3'-diaminobenzidine (DAB) to construct HCC models were divided into control group, cancer-inducing group and UTI intervention group. In each group, rat liver samples were collected at weeks 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22 and 24 respectively to study pathological distribution characteristics of oval cells in the process of carcinogenesis under optical microscope. Oval cells were separated by the methods of improved density gradient centrifugation and their structural characteristics were observed under optical microscope and electronic microscope respectively; the oval cells expressing C-kit and PCNA in the collected samples were observed by the methods of immunohistochemistry and image analysis and the expression of c-myc mRNA was also detected by reverse transcription polymerase chain reaction (RT-PCR).RESULTS: Oval cells proliferated firstly in the portal area then gradually migrated into hepatic parenchyma in the inducing group and intervention group. The oval cells distributed inside and outside the carcinoma nodes. The oval cells presented the characteristics of undifferentiated cells: a high ratio of nucleolus and cellular plasm and obvious nucleoli, rare organelle in plasm. Only a few mitochondria and endoplasmic reticulum and some villuslike apophysis on surface of cells could be seen. Cells stained with C-kit and PCNA antibody were mainly oval cells distributed in the portal area. The expression of cmyc mRNA increased with the progression of HCC.However, in the intervention group, UTI could retard its increase.CONCLUSION: Oval cells work throughout the development of HCC, and might play important roles in this process.c-mycgene may be a kind of promoter gene of HCC, and play a key role in hepatic injury and development of HCC.UTI could retard the occurrence of HCC.展开更多
AIM: To explore the mechanism of benign biliary stricture. METHODS: A model of trauma of bile duct was established in 28 dogs. The anastomosed tissues were resected and examined by light and electron microscopes on da...AIM: To explore the mechanism of benign biliary stricture. METHODS: A model of trauma of bile duct was established in 28 dogs. The anastomosed tissues were resected and examined by light and electron microscopes on day 3, in wk 1, 3 and mo 3, 6 after operation. CD68, TGF-β1 and α-SMA were examined by immunohistochemical staining, respectively. RESULTS: The mucosal epithelium of the bile duct was slowly recovered, chronic inflammation lasted for a long time, fibroblasts proliferated actively, extracellular matrix was over-deposited. Myofibroblasts functioned actively and lasted through the whole process. The expression of macrophages in lamina propria under mucosa, TGF-β1 in granulation tissue, fibroblasts and endothelial cells of blood vessels, oc-SMA in myofiroblasts were rather strong from the 1st wk to the 6th mo after operation. CONCLUSION: The type of healing occurring in bile duct belongs to overhealing. Myofibroblasts are the main cause for scar contracture and stricture of bile duct. High expressions of CD68, TGF-β1 and a-SMA are closely related to the active proliferation of fibroblasts, extracellular matrix over-deposition and scar contracture of bile duct.展开更多
AIM: To evaluate the characters, risks and benefits of laparoscopic cholecystectomy (LC) in cirrhotic portal hypertension (CPH) patients.METHODS: Altogether 80 patients with symptomatic gallbladder disease and CPH, in...AIM: To evaluate the characters, risks and benefits of laparoscopic cholecystectomy (LC) in cirrhotic portal hypertension (CPH) patients.METHODS: Altogether 80 patients with symptomatic gallbladder disease and CPH, including 41 Child class A,32 Child class B and 7 Child class C, were randomly divided into open cholecystectomy (OC) group (38 patients) and LC group (42 patients). The cohorts were well-matched for number, age, sex, Child classification and types of disease.Data of the two groups were collected and analyzed.RESULTS: In LC group, LC was successfully performed in 36 cases, and 2 patients were converted to OC for difficulty in managing bleeding under laparoscope and dense adhesion of Calot's triangle. The rate of conversion was 5.3%. The surgical duration was 62.6±15.2 min. The operative blood loss was 75.5±15.5 mL. The time to resume diet was 18.3±6.5 h. Seven postoperative complications occurred in five patients (13.2%). All patients were dismissed after an average of 4.6±2.4 d. In OC group, the operation time was 60.5±17.5 min. The operative blood loss was 112.5±23.5 mL. The time to resume diet was 44.2±10.5 h.Fifteen postoperative complications occurred in 12patients (30.0%). All patients were dismissed after an average of 7.5±3.5 d. There was no significant difference in operation time between OC and LC group. But LC offered several advantages over OC, including fewer blood loss and lower postoperative complication rate, shorter time to resume diet and shorter length of hospitalization in patients with CPH.CONCLUSION: Though LC for patients with CPH is difficult, it is feasible, relatively safe, and superior to OC.It is important to know the technical characters of the operation, and pay more attention to the meticulous perioperative managements.展开更多
AIM: To elucidate the mechanism by which somatostatin and its analogue exert the influence on liver fibrosis, and to investigate the mRNA expression of somatostatin receptors subtypes (SSTRs) and the distribution of s...AIM: To elucidate the mechanism by which somatostatin and its analogue exert the influence on liver fibrosis, and to investigate the mRNA expression of somatostatin receptors subtypes (SSTRs) and the distribution of somatostatin analogue octreotide in rat hepatic stellate cells (HSCs). METHODS: HSCs were isolated from Sprague Dawley (SD) rats by in situ perfusion and density gradient centrifugation. After several passages, the mRNA expression of 5 subtypes of SSTRs were assessed by reverse transcription-polymerase chain reaction (RT-PCR). HSCs were planted on coverslip and co-cultured with octreotide tagged by FITC. Then the distribution of FITC fluorescence was observed under laser scanning confocal microscope (LSCM) in 12-24 h. RESULTS: There were mRNA expression of SSTR2, SSTR3 and SSTR5 but not SSTR1 and SSTR4 in SD rat HSCs. The mRNA expression level of SSTR2 was significantly higher than that of other subtypes (P<0.01). FITC fluorescence of octreotide was clearly observed on the surface and in the cytoplasm, but not in the nuclei of HSCs under LSCM. CONCLUSION: The effect exerted by somatostatin and its analogues on HSCs may mainly depend on the expression of SSTR2, SSTR3 and SSTR5. Octreotide can perfectlycombine with HSCs, and thereby exerts its biological activity on regulating the characters of active HSCs. This provides a potential prevention and management against liver fibrosis.展开更多
基金Supported by the National Natural Science Foundation of China,No.30300337,30200278,30170919
文摘AIM: To investigate the expression of toll-like receptor 4(TLR4) and MD-2 gene and protein in Kupffer cells (KCs)and their role in ischemia-reperfusion (IR) injury of ratliver graft.METHODS: KCs were isolated at 0 (control group), 2, 12,24 h (IR group) following IR in rat liver graft, mRNA expressionof TLR4 and MD-2 was detected by RT-PCR analysis, proteinexpression of TLR4/MD-2 was detected by flow cytometric(FCM) analysis, and tumor necrosis factor-(~ (TNF-(~) levelin supernatant was measured by ELISA. Then isolated KCswere incubated with anti-TLR4 polyclonal antibody (anti-TLR4 group), and TNF-(~ level was measured again.RESULTS: The mRNA and protein expression of TLR4/MD-2 and the level of TNF-(~ in IR group increased significantlyat 2 h following IR, and reached the maximum at 12 h, andslightly decreased at 24 h, but were still significantly higherthan those in the control group (P<O.01). The expression ofthese factors was markedly decreased after anti-TLR4antibody treatment as compared with the IR group (P<O.01).CONCLUSION: Lipopolysaccharide (LPS) following IR canup-regulate TLR4/MD-2 gene and protein expression inKCs, and synthesize cytokine TNF-(~. Anti TLR4 antibodycan inhibit the production of TNF-(~ induced by LPS. TLR4and its partner molecule MD-2 may play an important rolein Kupffer cell activation and IR injury.
文摘AIM:The diagnosis of cholangiocarcinoma is often difficult,making management approaches problematic. A reliable serum marker for cholangiocarcinoma would be a useful diagnostic test. The aims of our study were to evaluate the usefulness of a serum CA19-9 determination in the diagnosis of cholangiocarcinoma.METHODS: We prospectively measured serum CA19-9 and CEA concentrations in patients with cholangiocarcinoma (n=35), benign biliary diseases (n=92), and healthy individuals (n=15). Serum CA19-9 and CEA concentrations were measured by an immunoradiometric assay without knowledge of the clinical diagnosis.RESULTS:The sensitivity of a CA19-9 value>37KU·L^-1 and a CEA value >22μg·L^-1 in diagnosing cholangiocarcinoma were 77.14% and 68.57%, respectively. When compared with the benign biliary diseases group,the true negative rates of serum CA19-9 and CEA were 84.78% and 81.52%,respectively. The false positive rates of serum CA19-9 and CEA were 15.22% and 18.48%, whereas the accuracy of serum CA19-9 and CEA were 82.68% and 77.95%,respectively. Serum CA19-9 and CEA concentrations were significantly elevated (P<0.001 and P<0.05) in patients with cholangiocarcinoma (290.31±5.34KU·L^-1 and 36.46±18.03μg·L^-1) compared with patients with benign biliary diseases (13.38±2.59KU·L^-1 and 13.84±3.85μg·L^-1) and healthy individuals (12.78±3.69KU·L^-1 and 11.48±3.37μg·L^-1). In 15 patients undergoing curative resection of cholangiocarcinoma,the mean serum CA19-9 concentration was decreased from a preoperative level of 286.41±4.36KU·L^-1 to a postoperative level of 62.01±17.43KU·L^-1 (P<0.001), and the mean serum CEA concentration from 39.41±24.35μg·L^-1 to 28.69±11.03μg·L6-1(P<0.05). In patients with cholangiocarcinoma,however, no correlation was found between serum CEA and CA19-9 concentrations (r=-0.036).CONCLUSION:These data suggest that the serum CA19-9 determination is a useful addition to the available tests for the differential diagnosis of cholangiocarcinoma. Serum CA19-9 is an effective tumor marker in diagnosing cholangiocarcinoma,deciding whether the tumor has been radically resected and monitoring effect of treatment.
基金Supported by Traditional Chinese Medicine Bureau Foundation of Shaanxi Province,No.2001-035
文摘AIM: To study the anti-tumor effect of resveratrol and in combination with 5-FU on murine liver cancer. METHODS: Transplantable murine hepatoma22 model was used to evaluate the anti-tumor activity of resveratrol (RES) alone or in combination with 5-FU in vivo. H22 cell cycles were analyzed with flow cytometry. RESULTS: Resveratrol could inhibit the growth of murine hepatoma22, after the mice bearing H22 tumor were treated with 10 mg/kg or 15 mg/kg resveratrol for ten days, and the inhibition rates were 36.3% (n = 10) and 49.3% (n = 9), respectively, which increased obviously compared with that in control group (85+22 vs 68+17, P<0.01). RES could induce the S phase arrest of H22 cells, and increase the persentage of cells in S phase from 59.1% (n = 9) to 73.5% (n = 9) in a dose-dependent manner (P<0.05). The enhanced inhibition of tumor growth by 5-FU was also observed in hepatoma22 bearing mice when 5-FU was administered in combination with 10 mg/kg resveratrol. The inhibition ratesfor 20 mg/kg or 10 mg/kg 5-FU in combination with 10 mg/kg resveratrol were 77.4% and 72.4%, respectively, compared with the group of 20 mg/kg or 10 mg/kg 5-FU alone, in which the inhibition rates were 53.4% and 43.8%, respectively (n = 8). There was a statistical significance between the combination group and 5-FU group. CONCLUSION: RES could induce the S phase arrest of H22 cells and enhance the anti-tumor effect of 5-FU on murine hepatoma22 and antagonize its toxicity markedly. These results suggest that resveratrol, as a biochemical modulator to enhance the therapeutic effects of 5-FU, may be potentially useful in cancer chemotherapy.
基金Supported by the National Natural Science Foundation of China, No.30371398
文摘AIM: To observe the effect of resveratrol on nuclear factor Kappa-B (NF-κB) activation and the inflammatory response in sodium taurocholate-induced pancreatitis in rats. METHODS: Seventy-two male SD rats were randomly divided into three groups: sham operation group (control), severe acute pancreatitis (SAP) group, and severe acute pancreatitis group treated with resveratrol (RES). A SAP model was established by injecting 4% sodium taurocholate 1 mL/kg through puncturing the pancreatic duct. In Res group, Res was given at 30 mg/kg b.m. intraperitoneally after the SAP model was successfully established. Eight animals from each group were sacrificed at 3, 6 and 12 h after modeling. The expression of NF-κB activation of pancreas was detected by irnmunohistochemical staining, whereas the levels of TNF-α and IL-8 in pancreatic tissues were estimated by radioimrnunoassay. The pathological changes of pancreas and lungs were examined microscopically. RESULTS: Much less hyperemia, edema, dust-colored necrotic focus and soaps were noticed in pancreas in RES group than in SAP group. In RES group, hemorrhage, exudates and infiltration of inflammatory cells in pancreas and interstitial edema, destruction of alveolar wall in lung were significantly less than in SAP group. In the SAP group, the activation of NF-κB in pancreatic tissues was enhanced significantly at any measure point compared with control group (64.23±10.72% vs 2.56±0.65%, 55.86±11.34% vs 2.32±0.42%, 36.23±2.30% vs 2.40±0.36%,P<0.01), TNF-α, IL-8 were also increased and reached their peak at 6 h and then declined. The activation of NF-κB and the levels of TNF-α and IL-8 in RES group were significantly lower than those in SAP group (P<0.01): activation (52.63±9.45% vs 64.23±10.72%, 40.52±8.40% vs 55.86±11.34%, 29.83±5.37% vs 36.23±2.30%), TNF-α (132.76±15.68 pg/mL vs 158.36±12.58 pg/mL, 220.32±23.57 pg/mL vs 247.67± 11.62 pg/mL, 175.68±18.43 pg/mL vs 197.35±12.57 pg/mL) and IL-8 (0.62±0.21 μg/L vs 0.83±0.10 μg/L, 1.10±0.124 μg/L vs1.32±0.18 μg/L, 0.98±0.16 μg/L vs 1.27±0.23μg/L). CONCLUSION: The activation of NF-KB is involved in the inflammatory response of rats with SAP. Resveratrol could effectively inhibit the expression of NF-κB activation, alleviate the severity of SAP through its anti-inflammatory effects and regulate the inflammatory mediators.
基金Supported by the National Natural Science Foundation of China,No.30070741
文摘AIM: To study the protective effects of tumor necrosis factor α (TNFα) antibody and ulinastatin on liver ischemic reperfusion in rats.METHODS: One hundred and twenty male SD rats were randomly divided into four groups: normal control group, ischemic group, TNFα antibody group and TNFα antibody + ulinastatin group. The animals were killed at 0, 3, 6, 9, 12 h after ischemia for 60 min and followed by reperfusion. Serum alanine aminotransferase (ALT), malondialdehyde (MDA) and liver histopathology were observed.RESULTS: After ischemic reperfusion, the serum ALT and MDA were remarkably increased, and the hepatic congestion was obvious. Treatment of TNFα antibody and ulinastatin could significantly decrease serum ALT and MDA levels, and relieve hepatic congestion.CONCLUSION: Ulinastatin and TNFα antibody can suppress the inflammatory reaction induced by hepatic ischemic reperfusion, and have protective effects on rat hepatic ischemic reperfusion injury.
文摘AIM: To investigate the protective effect of ischemic preconditioning (IPC) on hepatocellular carcinoma (HCC) patients with cirrhosis undergoing hepatic resection under hepatic inflow occlusion (HIO) and its possible mechanism.METHODS: Twenty-nine consecutive patients with resectable OHCC were randomized into two groups: IPC group: before HIO, IPC with 5 min of ischemia and 5 min of reperfusion was given; control group: no IPC was given. Uver functions,hepatic Caspase-3 activity, and apoptotic cells were compared between these two groups.RESULTS: On postoperative days (POD) 1, 3 and 7, the aspartate transaminase (AST) and alanine transaminase (ALT) levels in the IPC group were significantly lower than those in the control group (P<0.05). On POD 3 and 7, the total bilirubin level in the IPC group was significantly lower than that in the control group (P<0.05). On POD 1, the albumin level in the IPC group was higher than that in the control group (P = 0.053). After 1 h of reperfusion, both hepatic Caspase-3 activity and apoptotic sinusoidal endothelial cells in the IPC group were significantly lower than those in the control group (P<0.05).CONCLUSION: IPC has a potential protective effect on HCC patients with cirrhosis. Its protective mechanism underlying the suppression of sinusoidal endothelial cell apoptosis is achieved by inhibiting Caspase-3 activity.
文摘AIM: To study changes in characteristics of colorectal carcinoma during the metastatic process and to investigate the correlation between cell proliferation activity and metastatic ability of patients with Dukes′ stage C or D.METHODS: Formalin fixed and paraffin embedded materials of primary tumors and corresponding lymph node metastases resected from 56 patients with Dukes′ stage C or D of colorectal carcinoma were stained immunohistochemically with proliferating cell nuclear antigen (PCNA) and CD44variant exon 6 (CD44v6).RESULTS: Thirty-one of 56 patients (55.4 %) expressed PCNA in the primary sites and 36 of 56 patients (64.3 %)expressed PCNA in the metastatic lymph nodes. A significant relation in PCNA expression was observed between the primary site and the metastatic lymph node (0.010＜P＜0.025).Forty-one of 56 patients (73.2 %) expressed CD44v6 in the primary site and 39 of 56 patients (69.6 %) expressed CD44v6 in the metastatic lymph node. There was also an significant relationship of CD44v6 between the primary site and the metastatic lymph node (0.005＜P＜0.010). No difference was observed between expression of CD44v6 and PCNA in the primary site (0.250＜P＜0.500).CONCLUSION: This study partially demonstrates that tumor cells in metastatic lymph node of colorectal carcinoma still possess cell proliferation activity and metastatic ability of tumor cells in primary site. There may be no association between cell proliferation activity and metastatic ability in colorectal carcinoma.
文摘AIM: To evaluate the effects of nitroester drugs on human sphincter of Oddi (SO) motility and their antagonistic effects against morphine which shows excitatory effect on Oddi's sphincter motility.METHODS: The effects of these drugs on SO were evaluated by means of choledochofiberoscopy manometry.A total of 67 patients having T-tubes after cholecystectomy and choledochotomy were involved in the study, they were randomly divided into glyceryl trinitrate (GTN) group,isosorbide dinitrate (ISDN) group, pentaerythritol tetranitrate (PTN) group, morphine associated with GTN group, morphine associated with ISDN group and morphine associated with PTN group. Basal pressure of Oddi's sphincter (BPOS), amplitude of phasic contractions (SOCA), frequency of phasic contractions (SOF), duration of phasic contractions (SOD), duodenal pressure (DP) and common bile duct pressure (CBDP) were scored and analyzed. Morphine was given intramuscularly while nitroester drugs were applied sublingually.RESULTS: BPOS and SOCA decreased significantly after administration of ISDN and GTN, BPOS reduced from 10.95±7.49 mmHg to 5.92±4.04 mmHg (P＜0.05) evidently after application of PTN. BPOS increased from 7.37±5.58mmHg to 16.60±13.87 mmHg, SOCA increased from 54.09±38.37 mmHg to 100.70±43.51 mmHg, SOF increased from 7.15±3.20 mmHg to 10.38±2.93 mmHg and CBDP increased 3.75±1.95 mmHg to 10.49±8.21 mmHg (P＜0.01)evidently after injection of morphine. After associated application of ISDN and GTN, the four indications above decreased obviously. As for application associated with PTN,SOCA and SOF decreased separately from 100.64±44.99mmHg to 66.17±35.88 mmHg and from 10.70±2.76 mmHg to 9.04±1.71 mmHg (P＜0.05) markedly.CONCLUSION: The regular dose of GTN, ISDN and PTN showed inhibitory effect on SO motility, morphine showed excitatory effect on SO while GTN, ISDN and PTN could antagonize the effect of morphine. Among the three nitroester drugs, the effect of ISDN on SO was most significant.
文摘AIM:Intrahepatic extension is the main cause of liver failure and death in hepatocellular carcinoma patients.The small GTPase Rho and one of its effector molecules ROCK regulate cytoskeleton and actomyosin contractility,and play a crucial role in cell adhesion and motility.We investigated the role of small GTPase Rho in biological behaviors of hepatocellular carcinoma to demonstrate the importance of Rho in cancer invasion and metastasis.METHODS:Using Western blotting,we quantitated Rho protein expression in SMMC-7721 cells induced by Lysophosphatidic acid (LPA).Furthermore,we examined the role of Rho signaling in regulating the motile and invasive properties of tumor cells.RESULTS:Rho protein expression was stimulated by LPA.Using the Rhotekin binding assay to assess Rho activation,we observed that the level of GTP-bound Rho was elevated transiently after the addition of LPA,and Y-27632 decreased the level of active Rho.LPA enhanced the motility of tumorcells and facilitated their invasion.Rho played an essential role in the migratory process,as evidenced by the inhibition of migration and motility of cancer cells by a specific inhibitor of ROCK,Y-27632.CONCLUSION: The finding that invasiveness of hepatocellular carcinoma is facilitated by the Rho/Rho-kinase pathway is likely to be relevant to tumor progression and Y-27632 may be a new potential effective agent for the prevention of intrahepatic extension of human liver cancer.
文摘AIM: To assess the effects of intramuscular analgesics(morphine, Ap-237, pethidine and tramadol) on humanOddi's sphincter motility with choledochoscope manometry.METHODS: A total of 70 patients having T tubes aftercholecystectomy and choledochotomy were assessed bycholedochoscope manometry. They were randomly dividedinto morphine group, Ap-237 group, pethidine group andtramadol group. Basal pressure of Oddi's sphincter (BPOS),amplitude of phasic contractions (SOCA), frequency ofphasic contractions (SOF), duration of phasic contractions(SOD), duodenal pressure (DP) and common bile ductpressure (CBDP) were scored and analyzed. All narcoticanalgesic drugs were administered intramuscularly.RESULTS: Levels of BPOS, SOCA and SOF were increasedafter injection of morphine and Ap-237 (P<0.05), level of CBDPwas increased from 4.97+3.87 mmHg to 8.62+7.43 mmHg(10 min later) and 7.32+5.95 mmHg (20 min later) afterinjection of morphine (P<0.01). No apparent changeoccurred after intramuscular injection of pethidine. Level ofBPOS was increased from 7.01+5.50 mmHg to 2.87+2.78 mmHg10 min after injection of tramadol and SOCA was decreasedfrom 63.34+35.29 mmHg to 45.90+27.86 mmHg (10 minlater, P<0.05) and 35.97+24.30 (20 min later, P<0.01) afteradministration of tramadol.CONCLUSION: All these findings indicate that Oddi'ssphincter manometry via choledochoscope is a practicaland new way to study the dynamics of Oddi' s sphincter.The regular dose of morphine and Ap-237 could increaseBPOS, SOF and SOCA. Morphine could increase the levelof CBDP, demonstrating an excitatory effect on the sphincterof Oddi. Pethidine had no effect on Oddi's sphincter motility.Tramadol shows an inhibitory effect on the motility of thesphincter of Oddi and decreases levels of BPOS and SOCA.
基金the Chinese Medicine Administration Bureau of Jiangsu Province,No SZ 9902
文摘AIM:To study the effect of matrine on activation of Kupffer cell during cold ischemia and reperfusion injury in rat orthotopic liver transplantation(OLT).
基金Supported by the National Natural Science Foundation of China, No. 30371398
文摘AIM: To investigate the mechanism of resveratrol underlying the microcirculation disorder and lung injury following severe acute pancreatitis (SAP). METHODS: Twenty-four rats were divided into 3 groups (SAP, sham and resveratrol groups) randomly. SAP model was established by injecting 4% sodium taurocholate l mL/kg through puncturing pancreatic ducts. Sham (control) group (8 rats) was established by turning over the duodenum. Resveratrol was given at 0.1 mg/kg b.m. intraperitoneally. Rats were sacrificed 9 h after SAP was induced. Blood samples were obtained for hemorrheological examination. Lung tissues were used for pathological observation, and examination of microvascular permeability, dry/wet ratio and myeloperoxidase (MPO) activity. Gene expression of intercellular adhesion molecule-1 (ICAM-1) was detected by RT-PCR. RESULTS: Compared with SAP group, resveratrol relieved the edema and infiltration of leukocytes in the lungs. Resveratrol improved markers of hemorrheology: high VTB (5.77±1.18 mPas vs9.49±1.34 mPas), low VTB (16.12±3.20 mPas vs30.91±7.28 mPas), PV (4.69±1.68 mPas vs 8.00±1.34 mPas), BSR (1.25±0.42 mm/h vs50.03±0.03 mm/h), VPC (54.67±3.08% vs 62.17±3.39%), fibrinogen (203.2?7.8 g/ L vs 51.3±19.1 g/L), original hemolysis (0.45±0.02 vs 0.49±0.02), and complete hemolysis (0.41±0.02 vs 0.43±0.02) (P<0.05). Resveratrol decreased the OD ratio of ICAM-1 gene (0.800±0.03 vs 1.188±0.10), dry/wet ratio (0.74±0.02 vs 0.77±0.03), microvascular permeability (0.079±0.006 vs 0.112±0.004) and MPO activity (4.42±0.32 vs 5.03±0.51) significantly (P<0.05). CONCLUSION: Resveratrol can improve the microcirculation disorder of the lung by decreasing leukocyte-endothelial interaction, reducing blood viscosity, improving the decrease of blood flow, and stabilizing erythrocytes in SAP rats. It may be a potential candidate to treat SAP and its severe complications (ALI).
文摘AIM: To sum up the clinical characteristics of patients with central pontine myelinolysis (CPM) after orthotopic liver transplantation (OLT) and to document the possible causes of CPM.METHODS: Data of 142 patients undergoing OLT between January 1999 to May 2003 were analyzed retrospectively.Following risk factors during perioperation were analyzed in patients with and without CPM: primary liver disease,preoperative serum sodium level, magnesium level and plasma osrnolality, fluctuation degree of serum sodium concentration, and irnmunosuppressive drug level, etc.RESULTS: A total of 13 (9.2%) neurologic symptoms appeared in 142 patients post-operation including 5 cases (3.5%) with CPM and 8 cases (5.6%) with cerebral hemorrhage or infarct. Two patients developing CPM after OLT had a hyponatrernia history before operation (serum sodiurn<130 mmol/L), their mean serum sodium level was 130.6±5.54 mmol/L. The serum sodium level was significantly lower in CPM patients than in patients without neurologic complications or with cerebral hemorrhage/infarct (P<0.05).The increase in serum sodiurnduring perioperative 48 h after OLT in patients wibh CPM was significantly greater than bhat in patients with cerebral hemorrhage/infarct but without neurologic complications (19.5+6.54 retool/L, 10.1+6.43 retool/L,4.5±4.34 mmol/L, respectively, P<0.05). Plasma osrnolality was greatly increased postoperation in patients with CPM.Hypomagnesernia was noted in all patients perioperation,but there were no significant differences between groups.The duration of operation on patients with CPM was longer than that on others (492+190.05 rnin, P<0.05). Cyclosporin A (CsA) levels were normal in all patients, but there were significant differences between patients with or without neurologic complications (P<0.05).CONCLUSION: CPM may be more prevalent following liver transplantation. Although the diagnosis of CPM after OLT can be made by overall neurologic evaluations including magnetic resonance imaging (MRI) of the head, the mortality is still very high. The occurance of CPM may be associated with such factors as hyponatremia, rapid rise of serum sodium concentration, plasma osmolality increase postoperation, the duration of operation, and high CsA levels.
基金Supported by the Education Foundation of Xuzhou Anesthesia Laboratory,Jiangsu,No.KJS02055
文摘AIM: To study the mechanism and effect of nuclear factor-κB (NF-κB) activation and inflammatory response on the extended cold-preserved graft injury after orthotopic liver transplantation (OLT). METHODS: OLT was performed in rats with varying time of cold ischemia grafts (6, 18 and 24 h in University Wisconsin solution at 4 ℃). We determined the time of NF-κB activation and expression of tumor necrosis factor-α (TNF-α), cytokineinducible neutrophil chemoattractant (CINC), and intercellular adhesion molecule-1 (ICAM-1) within 6 h after reperfusion. Serum alarming aminotransferase (ALT), neutrophil sequestration, circulating neutrophil CD11b and L- selectin expression were also evaluated. RESULTS: The accumulation of neutrophils in the graft was significantly increased in the 18 h and 24 h cold-ischemia groups within 0.5 h after reperfusion, compared with the 6 h group. But the strongly activated neutrophils was slightly increased at 2 h after reperfusion and remained at high levels 4 h after reperfusion, which was synchronized with the common situation of recipients after transplantation. Prolonged cold-preservation did not affect neutrophil accumulation and activation. NF-κB activation preceded the expression of TNF-α, CINC, and ICAM-1 in the liver, which was significantly increased with prolonged cold preservation. In prolonged cold preserved grafts, prominently elevated NF-κB activation occurred at 0.5 h and 1 h, compared with that at 2 h after reperfusion, which was consistent with greatly increased intrahepatic TNF-α response. CONCLUSION: NF-κB activation is correlated with the expression of TNF-α, CINC, and ICAM-1 in vivo in OLT rats. Extended cold preservation of grafts might up-regulate TNF-α, CINC, and ICAM-1 expression in the grafts, most probably through elevated NF-κB activation, and might contribute to neutrophil infiltration in the grafts after reperfusion. Elevated NF-κB activity is harmful to inflammatory response in the grafts, and inhibited NF-κB activity might protect against early graft injury after liver transplantation.
文摘AIM: To develop an in vitro three-dimensional (3-D) angiogenesis system to analyse the capillary sprouts induced in response to the concentration ranges of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) and to quantify their synergistic activity.METHODS: Microcarriers (MCs) coated with human microvascular endothelial cells (HMVECs) were embedded in fibrin gel and cultured in 24-well plates with assay media. The growth factors bFGF, or VEGF, or both were added to the system. The wells (n = 8/group) were digitally photographed and the average length of capillary-like sprouts (ALS) from each microcarrier was quantitated.RESULTS: In aprotinin-stabilized fibrin matrix, human microvascular endothelial cells on the MCs invaded fibrin,forming sprouts and capillary networks with lumina. The angiogenic effects of bFGF or VEGF were dose-clependent in bhe range from 10 to 40 ng/mL. At d 1, 10 ng/mL of bFGF and VEGF induced angiogenesis with an ALS of 32.13±16.6 μm and 43.75±27.92 μm, respectively, which were significantly higher than that of the control (5.88±4.45 μm, P<0.01),and the differences became more significant as the time increased. In addition, the combination of 10 ng/mL of bFGF and VEGF each induced a more significant effect than the summed effects of bFGF (10 ng/mL) alone and VEGF (10 ng/mL) alone when analyzed using SPSS system for general linear model (GLM) (P= 0.011), and bhat also exceeded the effects by 20 ng/mL of either bFGF or VEGF.CONCLUSION: A microcarrier-based in vitro threedimensional angiogenesis model can be developed in fibrin.It offers a unique system for quantitative analysis of angiogenesis. Both bFGF and VEGF exert their angiogenic effects on HMVECs synergistically and in a dose-dependent manner.
文摘AIM:To explore the relationship between angiogenesis and biological behaviors of primary gallbladder carcinoma (PGBC),the relationship between the expression of inducible nitric oxide synthase (iNOS) and biological behaviors of PGBC and its relationship with the expression of iNOS and angiogenesis of PGBC.METHODS: The expression of iNOS and micro-vessel density (MVD) were assessed by immunohistochemical method and image analysis system in 40 specimens of PGBC and in 8 specimens of normal gallbladder. The immunostaining results and related clinicopathologic materials were analyzed by statistical methods.RESULTS: MVD in PGBC was significantly higher than that in normal gallbladder tissue (46±14 vs 14±6, P<0.05), and was not related with age, gender, tumor size and histological type. MVD of poorly and undifferentiated tumor tissues was higher than that of moderately-differentiated and well-differentiated tumor tissues (52±9 vs43±9 vs33±6, P<0.01).MVD of Nevin IV and V stages was higher than that of NevinI, Ⅱ and Ⅲ stages (52±8 vs 37±13, P<0.01). MVD of cases with lymphatic or liver metastasis was significantly higher than that without liver metastasis (55±6 vs 42±10, P<0.05) or lymphatic metastasis (53±8 vs38±8, P<0.01). The positive level index (PLI) of iNOS in PGBC was 0.435±0.134, and was not related with age, gender, tumor size, histological type,differentiation and clinical stage of PGBC. The PLI of iNOS in cases with lymphatic metastasis was higher than that without lymphatic metastasis (0.573±0.078 vs 0.367±0.064,P<0.01). The PLI of iNOS in cases with liver metastasis was higher than that without liver metastasis (0.533±0.067 vs0.424±0.084, P<0.05). There was a significant correlation between PLI of iNOS and MVD in PGBC (P<0.05).CONCLUSION:Angiogenesis of PGBC is significantly related to the biological behaviors of PGBC. The expression of iNOS is related to the biological behaviors of PGBC. The detection of MVD and the expression of iNOS in PGBC can be used as parameters to determine the degree of malignancy and prognosis.
基金Supported by the Natural Science Foundation of Guangdong Province,No.2001.010593,2002.020097 and Key Science and Technology Research Project of Guangzhou,No.B012000-X-005-01-05
文摘AIM: To study oval cells' pathological characteristics and relationship with the occurrence of hepatocellular carcinoma (HCC); to observe the form and structural characteristics of oval cells; to explore the expression characteristics of C-kit, PCNA mRNA and c-mycgene during the occurrence and development of HCC and the effect of ulinastatin (UTI) on C-kit and PCNA expression.METHODS: One hundred and twenty-five SD rats fed on 3,3'-diaminobenzidine (DAB) to construct HCC models were divided into control group, cancer-inducing group and UTI intervention group. In each group, rat liver samples were collected at weeks 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22 and 24 respectively to study pathological distribution characteristics of oval cells in the process of carcinogenesis under optical microscope. Oval cells were separated by the methods of improved density gradient centrifugation and their structural characteristics were observed under optical microscope and electronic microscope respectively; the oval cells expressing C-kit and PCNA in the collected samples were observed by the methods of immunohistochemistry and image analysis and the expression of c-myc mRNA was also detected by reverse transcription polymerase chain reaction (RT-PCR).RESULTS: Oval cells proliferated firstly in the portal area then gradually migrated into hepatic parenchyma in the inducing group and intervention group. The oval cells distributed inside and outside the carcinoma nodes. The oval cells presented the characteristics of undifferentiated cells: a high ratio of nucleolus and cellular plasm and obvious nucleoli, rare organelle in plasm. Only a few mitochondria and endoplasmic reticulum and some villuslike apophysis on surface of cells could be seen. Cells stained with C-kit and PCNA antibody were mainly oval cells distributed in the portal area. The expression of cmyc mRNA increased with the progression of HCC.However, in the intervention group, UTI could retard its increase.CONCLUSION: Oval cells work throughout the development of HCC, and might play important roles in this process.c-mycgene may be a kind of promoter gene of HCC, and play a key role in hepatic injury and development of HCC.UTI could retard the occurrence of HCC.
文摘AIM: To explore the mechanism of benign biliary stricture. METHODS: A model of trauma of bile duct was established in 28 dogs. The anastomosed tissues were resected and examined by light and electron microscopes on day 3, in wk 1, 3 and mo 3, 6 after operation. CD68, TGF-β1 and α-SMA were examined by immunohistochemical staining, respectively. RESULTS: The mucosal epithelium of the bile duct was slowly recovered, chronic inflammation lasted for a long time, fibroblasts proliferated actively, extracellular matrix was over-deposited. Myofibroblasts functioned actively and lasted through the whole process. The expression of macrophages in lamina propria under mucosa, TGF-β1 in granulation tissue, fibroblasts and endothelial cells of blood vessels, oc-SMA in myofiroblasts were rather strong from the 1st wk to the 6th mo after operation. CONCLUSION: The type of healing occurring in bile duct belongs to overhealing. Myofibroblasts are the main cause for scar contracture and stricture of bile duct. High expressions of CD68, TGF-β1 and a-SMA are closely related to the active proliferation of fibroblasts, extracellular matrix over-deposition and scar contracture of bile duct.
文摘AIM: To evaluate the characters, risks and benefits of laparoscopic cholecystectomy (LC) in cirrhotic portal hypertension (CPH) patients.METHODS: Altogether 80 patients with symptomatic gallbladder disease and CPH, including 41 Child class A,32 Child class B and 7 Child class C, were randomly divided into open cholecystectomy (OC) group (38 patients) and LC group (42 patients). The cohorts were well-matched for number, age, sex, Child classification and types of disease.Data of the two groups were collected and analyzed.RESULTS: In LC group, LC was successfully performed in 36 cases, and 2 patients were converted to OC for difficulty in managing bleeding under laparoscope and dense adhesion of Calot's triangle. The rate of conversion was 5.3%. The surgical duration was 62.6±15.2 min. The operative blood loss was 75.5±15.5 mL. The time to resume diet was 18.3±6.5 h. Seven postoperative complications occurred in five patients (13.2%). All patients were dismissed after an average of 4.6±2.4 d. In OC group, the operation time was 60.5±17.5 min. The operative blood loss was 112.5±23.5 mL. The time to resume diet was 44.2±10.5 h.Fifteen postoperative complications occurred in 12patients (30.0%). All patients were dismissed after an average of 7.5±3.5 d. There was no significant difference in operation time between OC and LC group. But LC offered several advantages over OC, including fewer blood loss and lower postoperative complication rate, shorter time to resume diet and shorter length of hospitalization in patients with CPH.CONCLUSION: Though LC for patients with CPH is difficult, it is feasible, relatively safe, and superior to OC.It is important to know the technical characters of the operation, and pay more attention to the meticulous perioperative managements.
基金Supported by National Nature Science Foundation of China,No.30271270
文摘AIM: To elucidate the mechanism by which somatostatin and its analogue exert the influence on liver fibrosis, and to investigate the mRNA expression of somatostatin receptors subtypes (SSTRs) and the distribution of somatostatin analogue octreotide in rat hepatic stellate cells (HSCs). METHODS: HSCs were isolated from Sprague Dawley (SD) rats by in situ perfusion and density gradient centrifugation. After several passages, the mRNA expression of 5 subtypes of SSTRs were assessed by reverse transcription-polymerase chain reaction (RT-PCR). HSCs were planted on coverslip and co-cultured with octreotide tagged by FITC. Then the distribution of FITC fluorescence was observed under laser scanning confocal microscope (LSCM) in 12-24 h. RESULTS: There were mRNA expression of SSTR2, SSTR3 and SSTR5 but not SSTR1 and SSTR4 in SD rat HSCs. The mRNA expression level of SSTR2 was significantly higher than that of other subtypes (P<0.01). FITC fluorescence of octreotide was clearly observed on the surface and in the cytoplasm, but not in the nuclei of HSCs under LSCM. CONCLUSION: The effect exerted by somatostatin and its analogues on HSCs may mainly depend on the expression of SSTR2, SSTR3 and SSTR5. Octreotide can perfectlycombine with HSCs, and thereby exerts its biological activity on regulating the characters of active HSCs. This provides a potential prevention and management against liver fibrosis.
