AIM: To investigate the expression level of plasma vascularendothelial growth factor (P-VEGF) in patients withhepatocellular carcinoma (HCC) and its relationship withthe clinicopathologic characteristics, and to exami...AIM: To investigate the expression level of plasma vascularendothelial growth factor (P-VEGF) in patients withhepatocellular carcinoma (HCC) and its relationship withthe clinicopathologic characteristics, and to examine thechanges of P-VEGF in the course of transcatheter arterialchemoembolization (TACE).METHODS: Peripheral blood samples were taken from 45HCC patients before and 1, 3, 7 d, and 1 mo after TACE.Plasma VEGF level was measured with the quantitativesandwich enzyme-linked immunosorbent assay (ELISA).Twenty patients with benign liver lesions and 17 healthycontrol subjects were also included in this study.RESULTS: Plasma VEGF levels in HCC patients weresignificantly elevated as compared to those in patients withbenign liver lesions (P = 0.006) and in the normal controls(P = 0.003). Significant differences were observed whenP-VEGF was categorized by tumor size (P = 0.006), portalvein thrombosis (P= 0.011), distant metastasis (P= 0.017),arterial-portal vein shunting (P = 0.026), and InternationalUnion Against Cancer (UICC) TNM stage (P = 0.044). Therewas no correlation between plasma level of VEGF and thelevel of alpha fetoprotein (^-FP) (r = 0.068, P = 0.658) andweakly correlated with the number of platelets (r = 0.312,P = 0.038). P-VEGF levels increased significantly andreached the peak value on the first day after TACE, and thendecreased gradually. The change rate of P-VEGF concentration(one month post-TACE/pre-TACExl00%) was correlatedwith the retention rate of lipiodol oil (rs = 0.494, P= 0.001)and the tumor volume change (r s = 0.340, P = 0.034).The patients who achieved a partial or complete responseto TACE therapy showed significantly less pre-treatmentP-VEGF than those nonresponders (P = 0.025). A high pre-therapeutic P-VEGF level was associated with poor responseto treatment (P = 0.018).CONCLUSION: A high pre-treatment P-VEGF level is auseful marker for tumor nroeression, esBeciallv for vascularinvasion. TACE increases the level of P-VEGF onlytemporarily which may be associated with tumor ischemia.P-VEGF may be useful in predicting treatment response,monitoring disease course after TACE and judging the effectof different TACE regimens.展开更多
AIM: To investigate the effect of nutritional support therapy on severe acute pancreatitis (SAP).METHODS: A total of 96 patients with severe acute pancreatitis were divided randomly into control and treatment groups.T...AIM: To investigate the effect of nutritional support therapy on severe acute pancreatitis (SAP).METHODS: A total of 96 patients with severe acute pancreatitis were divided randomly into control and treatment groups.The former group received total parenteral nutrition (TPN)via central venous infusion, while parenteral nutrition (PN)and enteral nutrition (EN) therapies were applied in different phases for the latter group. The nutrition status, acute phase responses, pancreas lesions, enteric mucosa penetrability and immune functions were monitored.RESULTS: Body weight and prealbumin concentration were increased in treatment group, compared to those in the control group, but albumin concentration did not change significantly.Acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ)scores decreased after 7 d of treatment, whereas the scores of the control group decreased on the 11th day. Concentrations of tumor necrosis factor-α (TNF-α), interleukine-6 (IL-6) and serum C reactive protein (CRP) dropped earlier in the treatment group (on the 4th day) than that in the control group (on the 7th day). No difference was observed in pancreatic lesions between the control and treatment groups.Concentration of endotoxin and lactulose/manicol (L:M) ratio of urine did not change in treatment group, but those in the control group were elevated markedly. Compared with the treatment group, CD4:CD8 T cells ratio and immunoglobulin G (IgG) concentration in the control group decreased significantly.CONCLUSION: Compared to TPN, the combined therapy of EN and PN could improve the nutrition status and moderate the acute phase response obviously. Moreover, the integrity of enteric mucosa and immune function were protected more effectively in treatment group than in the control one. On the other hand, EN did not simulate the excretion of pancreas and avoid exaggerating the inflammation of pancreas. Thus,appropriate application of PN and EN appears to be more effective for patients with SAP.展开更多
AIM: To study the effects of pentoxifylline (PTX) on thecontent of hepatic TGF-β1, type Ⅰ and type Ⅲ collagen inschistosomiasis japonica mice with liver fibrosis and itsmechanism of anti-fibrosis.METHODS: Forty mic...AIM: To study the effects of pentoxifylline (PTX) on thecontent of hepatic TGF-β1, type Ⅰ and type Ⅲ collagen inschistosomiasis japonica mice with liver fibrosis and itsmechanism of anti-fibrosis.METHODS: Forty mice with schistosomiasis were dividedinto four groups: one group as control without anytreatment, other three were treated with Praziquantel 500mg/(kg.d)for 2 d, high dose PTX 360 mg/(kg.d) for 8 wk,and low dose PTX 180 mg/(kg.d) for 8 wk respectively.Immunohistochemical technique and multimedia colorpathographic analysis system were applied to observe thecontent change of hepatic TGF-β1, type Ⅰ and type Ⅲcollagen in schistosomiasis japonica mice with liver fibrosisbefore and after PTX treatment.RESULTS: Effects of PTX on the content change of hepaticTGF-β1, type Ⅰ and type Ⅲ collagen in schistosomiasis japonicamice with liver fibrosis were related to the dosage of PTX,high dose PTX treated group could significantly reduce thecontent of TGF-β1 (0.709±0.111), type Ⅰ (0.644±0.108) andtype Ⅲ (0.654±0.152) collagen compared with those ofcontrol group (0.883±0.140, 0.771±0.156, 0.822±0.129)with statistical significance (P＜0.05). Low dose PTX couldalso reduce the hepatic content of TGF-β1 (0.752±0.152),type Ⅰ (0.733±0.117) and type Ⅲ (0.788±0.147) collagen,but without statistical significance (P＞0.05). Both high doseand low dose PTX groups have significant differences onthe content of TGF-β1, type Ⅰ and type Ⅲ collagen (P＜0.05,P＜0.05, P＜ 0.01,respectively).CONCLUSION: High dose of PTX treatment could reducethe content of hepatic TGF-β1, type Ⅰ and type Ⅲ collagensignificantly in schistosomiasis japonica mice with liverfibrosis, and thus plays its role of antifibrosis.展开更多
AIM: To study the effects of carbon dioxide on the metastatic capability of cancer cells, and to compare them with that of nitrogen.METHODS: The colon cancer cell CCL-228 was treated with 100 % carbon dioxide or nitro...AIM: To study the effects of carbon dioxide on the metastatic capability of cancer cells, and to compare them with that of nitrogen.METHODS: The colon cancer cell CCL-228 was treated with 100 % carbon dioxide or nitrogen at different time points and then cultured under normal condition. Twelve hours after the treatment, the survival rates of suspension cells and the expressions of e-cadherin and VEGF were examined.RESULTS: After 60 min of carbon dioxide and longer time of nitrogen treatment, the suspended cells increased and the expression of e-cadherin decreased while the expression of VEGF was enhanced significantly. And the effects of nitrogen were similar to, but weaker than, those of carbon dioxide.CONCLUSION: Carbon dioxide may improve the metastatic capability of cancer cells and its effects are significantly stronger than that of nitrogen. A sequential use of carbon dioxide and nitrogen in pneumoperitoneum may take the advantage of both gases.展开更多
AIM:To investigate the effects of autoantibodies against β_1-adrenoceptor in hepatitis virus myocarditis on action potential and L-type Ca^(2+) currents. METHODS:Fifteen samples of autoantibodies against β_1- adreno...AIM:To investigate the effects of autoantibodies against β_1-adrenoceptor in hepatitis virus myocarditis on action potential and L-type Ca^(2+) currents. METHODS:Fifteen samples of autoantibodies against β_1- adrenoceptor positive sera of patients with hepatitis virus myocarditis were obtained and IgGs were purified by octanoic acid extraction.Binding of autoantibodies against β_1- adrenoceptor to guinea pig cardiac myocytes was examined by immunofluorescence.Using the patch clamp technique, the effects on the action potential and I_(ca-L) of guinea pig cardiac myocytes caused by autoantibodies against β_1-adrenoceptor in the absence and presence of metoprolol were investigated. Cell toxicity was examined by observing cell morphology and permeability of cardiac myocytes to trypan blue. RESULTS:The specific binding of autoantibodies against β_1-adrenoceptor to guinea pig cardiomyocytes was observed. Autoantibodies against β_1-adrenoceptor diluted at 1:80 prolonged APO_(20),APD_(50)and APD_(90) by 39.2%,29.1% and 15.2% respectively,and only by 7.2%,5.3% and 4.1% correspondingly in the presence of 1 μmol/L metoprolol. Autoantibodies against β_1-adrenoceptor diluted at 1:80, 1:100 and 1:120 significantly increased the I_(ca-l) peak current amplitude at 0 mV by 55.87±4.39%,46.33±5.01% and 29.29±4.97% in a concentration-dependent manner.In contrast,after blocking of β_1-adrenoceptors (1 μmol/L metoprolol),autoantibodies against β_1-adrenoceptor diluted at 1:80 induced a slight increase of I_(ca-L) peak amplitude only by 6.81±1.61%.A large number of cardiac myocytes exposed to high concentrations of autoantibodies against β_1- adrenoceptor (1:80 and 1:100) were turned into rounded cells highly permeable to trypan blue. CONCLUSION:Autoantibodies against β_1-adrenoceptor may result in arrhythmias and/or impairment of myocardiums in HVM,which would be mediated by the enhancement of I_(ca-L.展开更多
AIM:To assess the vascularity of hepatocellular carcinoma (HCC) before and after transcatheter arterial chemoembolization (TACE) with the quantitative parameters obtained by first pass perfusion weighted MR imaging (F...AIM:To assess the vascularity of hepatocellular carcinoma (HCC) before and after transcatheter arterial chemoembolization (TACE) with the quantitative parameters obtained by first pass perfusion weighted MR imaging (FP-MRI). METHODS:Seventeen consecutive patients with one to three lesions in liver underwent FP-MRI before treatment. FP-MRI was also performed one,three,six,nine months, and one year after TACE.The baseline signal intensity (SO) of pre-TACE and one month after TACE was analyzed,the vascularity of HCC assessed by steepest slope of the signal intensity versus time curves (SS) was blindly correlated with their DSA feature and clinical outcome. RESULT:No significant difference was found on baseline signal intensity (S0) between pre-TACE and one month after TACE (F=0.309,P=0.583),The SS (mean,32% per second) of lesion one month after TACE was lower than that of pre-TACE (mean,69% per second),but with no statistical significance (F=3.067,P=0.092).When local recurrence occurred,the time intensity curves became steeper.The vascularity of HCC before and after TACE graded by SS closely correlated with that by DSA (K=0.453,P<0.05). CONCLUSION:FP-MRI is a useful criterion for selecting effective interventional treatment for patients with HCC in their initial treatment and during follow up.展开更多
AIM: To investigate the effects of hepatitis B virus x gene and its protein product HBxAg on apoptosis in hepatocyte line HL-7702. METHODS: The reconstituted plasmid pcDNA3-x was established through recombination DNA ...AIM: To investigate the effects of hepatitis B virus x gene and its protein product HBxAg on apoptosis in hepatocyte line HL-7702. METHODS: The reconstituted plasmid pcDNA3-x was established through recombination DNA technique; pcDNA3X was transfected into HL-7702 cells by lipid-mediated trasfection. Positive clones were screened by G418, and HL7702/HI3x cells were analysed by the RT-PCR to confirm the steady expression of X gene in HL-7702 cells. The apoptosis rate in HL-7702 cells was determined by flow cytometry, TUNEL technology, electronic microscope. At the mean time, pcDNA3-X was transfected transiently into HL-7702 cells, and total RNA from HL-7702 cells was extracted 24, 48, 72, 96 and 120 h after the transient transfection, and semiquantitative analysis was performed by RT-PCR to detect the expression of HI3V X gene. Furthermore, apoptosis rate in HL-7702 cells was determined by flow cytometry analysis at the different times. RESULTS: RT-PCR analysis showed that HI3V X gene could be expressed stably in HL-7702 cells. Both flow cytometry and TUNEL technology revealed that the apoptosis rates of HL-7702/HI3x cells were much higher than those of HL-7702/ pcDNA3 and HL-7702 cells. Furthermore, the apoptotic phenomena and apoptotic body were observed in HL-7702/ HI3x cells under electronic microscope, but not in HL-7702/ pcDNA3 and HL-7702 cells. In the experiment of transient transfection, RT-PCR reveals that X gene was expressed most at 72 h after transfection; and the apoptosis rate reached the highest at the same time. After that, the apoptosis rate was reduced with the decrease of the X gene expression. CONCLUSION: HBV X gene and X protein can promote the apoptosis in hepatocyte. And there exist a quantity-effect relationship between the X gene expression and apoptosis rate in hepatocyte.展开更多
AIM: To observe the change of tumor microcirculation after transcatheter arterial chemoembolization (TACE) with bletilla microspheres by using first pass perfusion MR imaging (FP) and Chinese ink casting.METHODS: VX2 ...AIM: To observe the change of tumor microcirculation after transcatheter arterial chemoembolization (TACE) with bletilla microspheres by using first pass perfusion MR imaging (FP) and Chinese ink casting.METHODS: VX2 carcinoma cells were surgically implanted into the left and right lobes of liver of 30 New Zealand white rabbits, which were divided into 3 groups at random. Emulsion of lipiodol mixed with mitomydn C, and 5-FU bletilla microspheres were injected into the hepatic artery respectively, and saline was used as control agent. MR imaging was performed with turbo-flash sequence 14 d after tumor implantation and 7 d after interventional therapy. The steepest slopes (SS) of the signal intensity versus time curves were created for quantitative analysis, 7.5% Chinese ink gelatin solution was injected through ascending artery (17 cases) or portal vein(2 cases) for lesion microvessel area (MVA) measurement after the last MRI examination.The correlation between perfusion imaging and MVA was studied blindly.RESULTS: The SS values at the rim of tumor in lipiodol group (mean, 49% per second) and bletilla group (mean,35% per second) were significantly decreased (P<0.05) as compared with control group (mean, 124% per second), no difference was found between lipiodol and bletilla groups(P>0.05). In lipiodol group, the MVAs (24 974±11 836μm^2) in the center of the tumor were significantly smaller than those of the control group (35 510±15 675 μm^2) (P<0.05),while the MVAs (80 031±22 745 μm^2) around the tumor were significantly increased because small and dense plexuses appeared around the tumor which correlated to intense reaction of granulation tissue. None of the vessels was seen in the tumor in bletilla group, the peripheral MVAs of the tumor were significantly smaller than those of the control group (P<0.05) and lipiodol group (P<0.05). There was a good correlation between SS and MVAs in control group (rsl, 0.985, P<0.0001) and bletilla group (rsl, 0.743,P<0.05), the correlation was not significant in lipiodol group(rsl, 0.527, P>0.05).CONCLUSION: TACE with bletilla microspheres may enhance its anti-tumor effect by inhibiting the angiogenesis,and FP-MRI provides useful information to assess the TACE effect by depicting tumor vascularization and perfusion,展开更多
AIM:Transcatheter arterial embolization (TAE) of the hepatic artery has been accepted as an effective treatment for unresectable hepatocellular carcinoma (HCC). However,embolized vessel recanalization and collateral c...AIM:Transcatheter arterial embolization (TAE) of the hepatic artery has been accepted as an effective treatment for unresectable hepatocellular carcinoma (HCC). However,embolized vessel recanalization and collateral circulation formation are the main factors of HCC growth and recurrence and metastasis alter TAE. Vascular endothelial growth factor (VEGF) plays an important role in tumor angiogenesis.This study was to explore the inhibitory effect of VEGF antisense oligodeoxynucleotides (ODNs) on VEGF expression in cultured Walker-256 cells and to observe the anti-tumor effect of intra-arterial infusion of antisense ODNs mixed with lipiodol on rat liver cancer.METHODS: VEGF antisense ODNs and sense ODNs were added to the media of non-serum cultured Walker-256 cells.Forty-eight hours later, VEGF concentrations of supernatants were detected by EUSA. Endothelial cell line ECV-304 cells were cultured in the supernatants. Seventy-two hours later,growth of ECV-304 cells was analyzed by NTT method. Thirty Walker-256 cell implanted rat liver tumor models were divided into 3 groups.0.2 mL lipiodol (LP group, n=10), 3OD antisense ODNs mixed with 0.2 mL lipiodol (LP+ODNs group, n=10) and 0.2 mL normal saline (control group, n=10) were infused into the hepatic artery. Volumes of tumors were measured by MRI before and 7 d alter the treatment.VEGF mRNA in cancerous and peri-cancerous tissues was detected by RT-PCR. Microvessel density (MVD) and VEGF expression were observed by immunohistochemistry.RESULTS: Antisense ODNs inhibited Walker-256 cells' VEGF expression, The tumor growth rate was significantly lower in LP+ODNs group than that in LP and control groups (140.1±33.8%, 177.9±64.9% and 403.9±69.4% respectively, F=60.019,P<0.01).VEGF mRNAs in cancerous and peri-cancerous tissues were expressed highest in LP group and lowest in LP+ODNs group. The VEGF positive rates showed no significant difference among LP, control and LP+ODNs groups (90%,70% and 50%, H=3.731, P>0.05).The MVD in LP+ODNs group (53.1±18.4) was significantly less than that in control group (73.2±20.4) and LP group(80.3±18.5) (F=5.44, P<0.05).CONCLUSION: VEGF antisense ODNs can inhibit VEGF expression of Walker-256 cells.It may be an antiangiogenesis therapy agent for malignant tumors. VEGF antisense ODNs mixed with lipiodol embolizing liver cancer is better in inhibiting liver cancer growth, VEGF expression and microvessel density than lipiodol alone.展开更多
AIM: TO investigate the effects of anti-Fas ribozyme on Fas expression and apoptosis in primary cultured mouse hepatocytes.METHODS: Mouse hepatocytes were isolated by using collagenase irrigation. A hammerhead ribozym...AIM: TO investigate the effects of anti-Fas ribozyme on Fas expression and apoptosis in primary cultured mouse hepatocytes.METHODS: Mouse hepatocytes were isolated by using collagenase irrigation. A hammerhead ribozyme targeting the Fas mRNA was constructed, and transfected into mouse hepatocytes via Effectene. Then Fas expression in mouse hepatocytes was detected by RT-PCR and western blotting.After being treated with anti-Fas antibody (JO2), hepatocytes viability was measured with MTI- assay. Caspase-3 proteolytic activity was detected, and cell apoptosis was measured according to Annexin V-FITC apoptosis detection kit.RESULTS: Fas expressed in primary mouse hepatocytes.Fas expression in hepatocytes transfected with anti-Fas ribozyme was decreased remarkably and correlated with the resistance to Fas-mediated apoptosis as determined by flow cytometry and caspase-3 proteolytic activity.CONCLUSION: AnU-Fas ribozyme can remarkably decrease the Fas expression in mouse hepatocytes, thus inhibit Fasmediated apoptosis in hepatocytes, It is suggested that anti-Fas ribozyme could significantly increase the resistance of transplanted hepatocytes to apoptosis and improve the survival of transplanted hepatocytes,展开更多
AIM: To investigate the effect of Kangxian ruangan keli (KXR) on hepatic stellate cell (HSC) proliferation mediated by platelet-derived growth factor (PDGF) and the underlying mechanism.METHODS: In a serum-free cultur...AIM: To investigate the effect of Kangxian ruangan keli (KXR) on hepatic stellate cell (HSC) proliferation mediated by platelet-derived growth factor (PDGF) and the underlying mechanism.METHODS: In a serum-free culture system, HSCs were treated with a KXR preparation for 24 hours, followed by stimulation with PDGF-BB for 24 hours. Then the cells were incubated again in the medium containing KXR for 3 hours stimulated with PDGF-BB for 5 minutes, and collected. The proliferation of HSC was examined using an MTT assay and flow cytometry. Tyrosine phosphorylation was detected with Western blotting and visualized by the enhenced chemiluminescent (ECL) method.RESULTS: The OD values for the HSCs growing in the media without and with addition of PDGF were 0.17±0.06 and0.82±0.05, respectively. The PDGF-induced increase was hindered remarkably by KXR preparation in a dose-dependent manner. The reaction values for the systems with 5mg/mL, 2.5 mg/mL and 1.25 mg/mL of KXR were 0.28±0.03,0.37±0.02 and 0.43±0.04, respectively. Moreover, the percentages of S-phase cells in these KXR-containing culturesystems were 10.95±1.35, 32.76±1.07 and 43.19±1.09,respectively, all of which were significantly lower than that in the culture free of KXR (68.24±2.72). In addition, the values for tyrosine-phosphorylated protein in HSCs treated with 5 mg/mL and 1.25 mg/mL of KXR were 0.1349±0.0072 and 0.1658±0.0025, respectively, which were smaller than that in the cells treated only with PDGF-BB (0.1813±0.0117).CONCLUSION: Within the dose range used in the present study, KXR preparation shows an inhibitory effect on HSC proliferation induced by PDGF. The mechanism of this process may involve interference with tyrosine phosphorylation mediated by PDGF.展开更多
AIM： To investigate the role of SMYD3 in hepatocellular carcinoma （HCC） development and progression and to verify whether its regulation activity was through RIZ1 inactivation. METHODS： Expression of SMYD3 in HCC ...AIM： To investigate the role of SMYD3 in hepatocellular carcinoma （HCC） development and progression and to verify whether its regulation activity was through RIZ1 inactivation. METHODS： Expression of SMYD3 in HCC cell lines and tissues were measured; silencing of SMYD3 by RNA interference （RNAi） was effectuated, hepatoma cell proliferation, migration and apoptosis were tested, with RIZl CpG promoter methylation, and corresponding mRNA expression were investigated. RESULTS： SMYD3 over-expression in HCC was associated with RIZl hypermethylation and mRNA down-expression. Suppression of SMYD3 expression de- methylated RIZl CpG promoter （P 〈 0.01） and increased RIZl mRNA expression （P 〈 0.01）. Consequently, SMYD3 down-expression with RIZl de-methylation strongly inhibited hepatoma cell growth （MTT inhibitory rates： Pgenesil-1-s1 60.95%± 7.97%, Pgenesil-1-s2 72.14% ± 9.68% vs Pgenesil-1-hk 6.89% ± 4.12%, P 〈 0.01） and migration （Pgenesil-1-s1 4.24% ± 1.58%, Pgenesil- 1-s1 4.87% ± 0.73% vs Pgenesil-1 19.03% ± 4.63%, Pgenesil-1-hk 19.95% ±5.21%, P 〈 0.01） and induced apoptosis （FCM subG1 phase Pgenesil-1-s1 19.07% ＋ 1.78%, Pgenesil-1-s2 17.68% ± 2.36% vs Pgenesil-1 0.47% ± 0.12%, Pgenesil-1-hk 1.46% ± 0.28%, P 〈 0.01. TUNEL-positive cells： Pgenesil-1-s1 40.24%± 5.18%, Pgenesil-1-s2 38.48% ± 4.65% vs Pgenesil-1 2.1B% - 1.34%, Pgenesil-1-hk 2.84%± 1.22%, P 〈 0.01） in HepG2 cells. CONCLUSION： These results demonstrate that SMYD3plays a critical role in the carcinogenesis and progression of HCC, The proliferation, migration induction and apoptosis inhibition activities of SMYD3 may be mediated through RIZl CpG promoter hypermethylation.展开更多
AIM: To study the distribution and stability of antisense oligodeoxynucleotide (ASODN) in Walker-256 cells and their distribution in liver, lung and kidney tissues after being infused alone or mixed with lipiodol via ...AIM: To study the distribution and stability of antisense oligodeoxynucleotide (ASODN) in Walker-256 cells and their distribution in liver, lung and kidney tissues after being infused alone or mixed with lipiodol via hepatic artery in a rat liver tumor model.METHODS: 5'-Isothiocyananate (FITC)-labeled vascular endothelial growth factor (VEGF) ASODN was added into Walker-256 cell culture media. Its distribution in cells was observed by fluorescence microscope at different time points. Walker-256 carcinosarcoma was transplanted into Wistar rat liver to establish a liver cancer model. 5'-FITC-labeled VEGF ASODN mixed with (mixed group, n = 6) or without (TAI group, n = 6)ultra-fluid lipiodol was administrated via hepatic artery.Frozen samples of liver, lung and kidney tissue were taken from rats after 1, 3 and 6 d, respectively. The distribution of ASODN was observed under fluorescent microscope.RESULTS: ASODN could enter cytoplasm within 2 h and nuclei within 6 h. Accumulation of ASODN reached the peak point in nuclei at 12 h, and then disappeared gradually. No fluorescence could be seen in cells at 48 h. In vivo experiment, on d 1 and 3 the fluorescence staining in liver was stronger in mixed group than in TAI group and more fluorescence could be detected in lung and kidney in TAI group than in mixed group. On d 6, no fluorescence could be detected in TAI group, but faint fluorescence could be seen in mixed group. ASODN could be seen in cancer cells and normal hepatic cells. In mixed group, ASODN was mainly distributed in liver tumor tissues.CONCLUSION: ASODN can transfect Walker-256 cells.ASODN mixed with lipiodol infusion via hepatic artery can be used in the treatment of HCC.展开更多
AIM: This study was designed to examine the hypothesis that gender differences in I/R injury are associated withendothelial cell nitric oxide synthase (eNOS)-derived nitric oxide (NO).METHODS: Wistar rats were randomi...AIM: This study was designed to examine the hypothesis that gender differences in I/R injury are associated withendothelial cell nitric oxide synthase (eNOS)-derived nitric oxide (NO).METHODS: Wistar rats were randomized into seven experimental groups (12 animals per group). Except for the sham operated groups, all rats were subjected to total liver ischemia for 40 min followed by reperfusion. All experimental groups received different treatments 45 min before the laparotomy. For each group, half of the animals (six) were used to investigate the survival; blood samples and liver tissues were obtained in the remaining six animals after 3 h of reperfusion to assess serum NO, alanine aminotransferase (ALT) and TNF-α levels, liver tissuemalondialdehyde (MDA) content, and severity of hepatic I/R injury.RESULTS: Basal serum NO levels in female sham operated (FS) group were nearly 1.5-fold of male sham operated (MS) group (66.7±11.0 μmol/L vs 45.3±10.1μmol/L, P＜0.01). Although serum NO levels decreased significantly after hepatic I/R (P＜0.01, vs sham operated groups), they were still significantly higher in female rat (F) group than in male rat (M) group (47.8±8.6 μmol/L vs 23.8±4.7 μmol/L, P＜0.01). Serum ALT and TNF-α levels, and liver tissue MDA content were significantly lower in F group than in M group (370.5±46.4 U/L, 0.99±0.11 μg/L and 0.57±0.10 μmol/g vs668.7±78.7 U/L, 1.71±0.18 μg/Land 0.86±0.11 μmol/g, respectively, P＜0.01). I/R induced significant injury to the liver both in M and F groups (P＜0.01 vs sham operated groups). But the degree of hepatocyte injury was significantly milder in F group than in M group (P＜0.05 and P＜0.01). The median survival time was six days in F group and one day in M group. The overall survival rate was significantly higher in F group than in M group (P＜0.05). When compared with male rats pretreated with saline (M group), pretreatment of male rats with 17-β- estradiol (E2) (M+E2 group) significantly increased serum NO levels and significantly decreased serum ALT and TNF-α levels, and liver tissue MDA content after I/R (P＜0.01).The degree of hepatocyte injury was significantly decreased and the overall survival rate was significantly improved in M+E2 group than in M group (P＜0.01 and P＜0.05). TheNOS inhibitor Nw-nitro-L-arginine methyl ester (L-NAME) treatment could completely abolish the protective effects of estrogen in both male and female rats. CONCLUSION: The protective effects afforded to female rats subjected to hepatic I/R are associated with eNOSderived NO.展开更多
AIM: To explore the cooperative effects of antisense oligonucleotide (ASON) of cell adhesion molecules and cimetidine on the expression of E-selectin and ICAM-1 in endothelial cells and their adhesion to tumor cells. ...AIM: To explore the cooperative effects of antisense oligonucleotide (ASON) of cell adhesion molecules and cimetidine on the expression of E-selectin and ICAM-1 in endothelial cells and their adhesion to tumor cells. METHODS: After treatment of endothelial cells with ASON and/or cimetidine and induction with TNF-α, the protein and mRNA changes of E-selectin and ICAM-1 in endothelial cells were examined by flow cytometry and RT-PCR,respectively. The adhesion rates of endothelial cells to tumor cells were measured by cell adhesion experiment. RESULTS: In comparison with TNF-α inducing group, lipoASON and lipo-ASON/cimetidine could significantly decrease the protein and mRNA levels of E-selectin and ICAM-1 in endothelial cells, and lipo-ASON/cimetidine had most significant inhibitory effect on E-selectin expression (from 36.37±1.56% to 14.23±1.07%, P<0.001). Meanwhile,cimetidine alone could inhibit the expression of E-selectin(36.37±1.56% vs 27.2±1.31%, P<0.001), but not ICAM-1(69.34±2.50% vs68.07±2.10%,P>0.05)and the two kinds ofmRNA, either. Compared with TNF-α inducing group, the rate of adhesion was markedly decreased in lipo-E-selectin ASON and lipo-E-selectin ASON/cimetidine treated groups(P<0.05),and lipo-E-selectin ASON/cimetidine worked better than lipo-E-selectin ASON alone except for HepG2/ECV304 group(P<0.05). However, the decrease of adhesion was not significant in lipo-ICAM-1 ASON and lipo-ICAM-1 ASON/dmetidine treated groups except for HepG2/ECV304 group (P >0.05). CONCLUSION: These data demonstrate that ASON in combination with cimetidine in vitro can significantly reduce the adhesion between endc^thelial cells and hepatic or colorectal cancer cells, which is stronger than ASON or cimetidine alone. This study provides some useful proofs for gene therapy of antiadhesion.展开更多
To determine the accuracy of triphase enhanced helical CT in judging the invasion and metastasis of gastric carcinoma, and to discuss the relation between imaging signs and pathological findings. Methods: Triphase enh...To determine the accuracy of triphase enhanced helical CT in judging the invasion and metastasis of gastric carcinoma, and to discuss the relation between imaging signs and pathological findings. Methods: Triphase enhanced helical CT scanning was performed in 46 patients with gastric carcinoma. Imaging findings were compared with postoperative pathologic results. Results: (1) The accuracy of helical CT for diagnosing involvement of tunica serosa, lymph node metastasis and distant metastasis was 84.8%, 87.0% and 100~ respectively. (2) CT signs of serosal involvement, lymph node metastasis and distant metastasis were in good accordance with pathological findings (P<0.05). Conclusion: Triphase enhanced helical CT scans can comprehensively and precisely reflect the pathologic characteristics of gastric carcinoma, thus it is a reliable technique for the diagnosis of this disease.展开更多
AIM: To study the effects of palmatine, a known inhibitoron delayed rectifier potassium current and L-type calciumcurrent (ICa,L) in guinea pig ventricular myocytes, on thepotassium and calcium currents in isolated ra...AIM: To study the effects of palmatine, a known inhibitoron delayed rectifier potassium current and L-type calciumcurrent (ICa,L) in guinea pig ventricular myocytes, on thepotassium and calcium currents in isolated rat hepatocytes.METHODS: Tight-seal wh ole-cell patch-clamp techniqueswere performed to investigate the effects of palmatine onthe delayed outward potassium currents (IK), inward rectifierpotassium current (IK1) and Ca2+ release-activated Ca2+current (ICRAC) in enzymatically isolated rat hepatocytes.RESULTS: Palmatine 0.3-100 μM reduced IK in a concentationdependent manner with EC50of 41.62±10.11 μM and nH,0.48±0.07 (n=8). The effect of the drug was poorly reversibleafter washout. When the bath solution was changed totetraethylammonium (TEA) 8 mM, IK was inhibited.Palmatine 10 μM and 100 μM shifted the I-V curves of IKdownward, and the block of IK was voltage-independent.Palmatine 0.3-100 μM also inhibited ICRAC in a concentration-dependent manner. The fitting parameters were as follows:ECs0=51.19±15.18 μM, and nH=0.46+0.07 (n=8). The peakvalue of ICRAC in the I-V relationship was decreased bypalmatine 10 μM and 100 μM. But the reverse potential ofIcRAcoCcurred at Voltage=0 mV in all cells. Palmatine 0.3-100 μM failed to have any significant effect on either inwardor outward components of IK1 at any membrane potentialexamined.CONCLUSION: The inhibitory effects on IK and ICRAC couldbe one of the mechanisms that palmatine exerts protectiveeffect on hepatocytes.展开更多
AIM: To examine the effects of berberine, an isoquinoline alkaloid with a long history used as a tonic remedy for liver and heart, on ion channels of isolated rat hepatocytes.METHODS: Tight-seal whole-cell patch-clamp...AIM: To examine the effects of berberine, an isoquinoline alkaloid with a long history used as a tonic remedy for liver and heart, on ion channels of isolated rat hepatocytes.METHODS: Tight-seal whole-cell patch-clamp techniques were performed to investigate the effects of berberine on the delayed outward potassium currents (Ik), inward rectifier potassium currents (Ik1) and Ca^2+ release-activated Ca^+ currents (ICRAC) in enzymatically isolated rat hepatocytes.RESULTS: Berbenne 1-300 μmol/L reduced/K in a concentration-dependent manner with EC50 of 38.86=1=5.37 μmol/L and nH of 0.82±0.05 (n = 8). When the bath solution was changed to tetraethylammonium (TEA) 8 retool/L,IK was inhibited.Berberine 30 μmol/L reduced/K at all examined membranepote ntials, especially at potentials positive to +60 mV (n = 8,P<0.05 or P<0.01 vs control). Berberine had mild inhibitory effects on IK1 in rat hepatocytes. Berberine 1-300 μmol/L also inhibited ICRAC in a concentration-dependent fashion. The fitting parameters were EC50 = 47.20±10.86 μmol/L,nH= 0.71±0.09 (n = 8). The peak value of/CRAC in the I-Vrelationship was decreased by berberine 30 μmol/L at potentialnegative to -80 mV (n = 8, P<0.05 vscontrol). But the reverse potential of/CRAC occurred at voltage 0 mV in all cells.CONCLUSION: Berberine has inhibitory effects on potassium and calcium currents in isolated rat hepatocytes, which may be involved in hepatoprotection.展开更多
To test the accuracy of real-time three-dimensional echocardiography (RT3DE) imaging system for evaluating left ventricular mass (LVM) in phantom and excised canine heart. Methods Ten left ventricular (LV) wall phanto...To test the accuracy of real-time three-dimensional echocardiography (RT3DE) imaging system for evaluating left ventricular mass (LVM) in phantom and excised canine heart. Methods Ten left ventricular (LV) wall phantoms made of two rubber-bursas, ten excised canine hearts underwent RT3DE and two-dimensional echocardiography (2DE). In RT3DE "full volume" imaging, the myocardial volume was mea-sured using 2,4,8, and 16-plane method with the analysis software of RT3DE. Mass was then calculated by multiplying the resulting myocardial volume by specific density of myocardial tissue. In 2DE the masses were measured by area-length meth-od. The true LV wall phantom mass was measured by water displacement and the canine LVM was weighed by anatomy, which served as a reference standard. We compared RT3DE or 2DE with true mass. Results In LV wall phantoms, RT3DE correlated with true masses strongly (r = 0.813-0.994) and weakly correlated between 2DE and true masses (r = 0.628). In excised canine hearts, there is an excellent correlation between RT3DE and true masses (r = 0.764-0.991), while 2DE value showed a lesser correlation (r = 0.514). There are no difference between RT-3DE and true masses (P > 0.05) but different between 2DE and true masses (P < 0.05). In different planes, there was no difference between 8-plane and 16-plane (P > 0.05) but different between 8-plane and 2, 4-plane (P < 0.05). Conclusion RT3DE can accurately quantify LVM and provide a new tool to evaluate LV function. For LVM by RT3DE, 8-plane measurement method is the best choice for accuracy and convenience.展开更多
We report two cases of extrahepatic portal vein aneurysm,and both of them underwent surgical intervention. The first case had a mild pain in right upper quadrant of the abdomen; the second had no obvious symptoms. Phy...We report two cases of extrahepatic portal vein aneurysm,and both of them underwent surgical intervention. The first case had a mild pain in right upper quadrant of the abdomen; the second had no obvious symptoms. Physical examination revealed nothing abnormal. Both of them were diagnosed by magnetic resonance imaging angiography (MRA). One of the aneurysms was located at the main portal vein, the other, at the confluence of the superior mesenteric vein and the splenic vein, and these two places are exactly the most common locations of the extrahepatic portal vein aneurysm reported in the literature (30.7% each site). The first case underwent aneurysmorrhaphy and the second case, aneurysm resection with splenectomy. Both of them recovered soon after the operation, and the symptom of the first case was greatly alleviated. During the follow-up of half a year, no complication and adverse effect of surgical intervention was found and the color Doppler ultrasonography revealed no recurrence of the aneurysmal dilation. We suggest that surgical intervention can alleviate the symptom of the extrahepatic portal vein aneurysm and prevent its complications effectively and safely for low risk patients.展开更多
基金Supported by the National Natural Science Foundation of China,No.39770839
文摘AIM: To investigate the expression level of plasma vascularendothelial growth factor (P-VEGF) in patients withhepatocellular carcinoma (HCC) and its relationship withthe clinicopathologic characteristics, and to examine thechanges of P-VEGF in the course of transcatheter arterialchemoembolization (TACE).METHODS: Peripheral blood samples were taken from 45HCC patients before and 1, 3, 7 d, and 1 mo after TACE.Plasma VEGF level was measured with the quantitativesandwich enzyme-linked immunosorbent assay (ELISA).Twenty patients with benign liver lesions and 17 healthycontrol subjects were also included in this study.RESULTS: Plasma VEGF levels in HCC patients weresignificantly elevated as compared to those in patients withbenign liver lesions (P = 0.006) and in the normal controls(P = 0.003). Significant differences were observed whenP-VEGF was categorized by tumor size (P = 0.006), portalvein thrombosis (P= 0.011), distant metastasis (P= 0.017),arterial-portal vein shunting (P = 0.026), and InternationalUnion Against Cancer (UICC) TNM stage (P = 0.044). Therewas no correlation between plasma level of VEGF and thelevel of alpha fetoprotein (^-FP) (r = 0.068, P = 0.658) andweakly correlated with the number of platelets (r = 0.312,P = 0.038). P-VEGF levels increased significantly andreached the peak value on the first day after TACE, and thendecreased gradually. The change rate of P-VEGF concentration(one month post-TACE/pre-TACExl00%) was correlatedwith the retention rate of lipiodol oil (rs = 0.494, P= 0.001)and the tumor volume change (r s = 0.340, P = 0.034).The patients who achieved a partial or complete responseto TACE therapy showed significantly less pre-treatmentP-VEGF than those nonresponders (P = 0.025). A high pre-therapeutic P-VEGF level was associated with poor responseto treatment (P = 0.018).CONCLUSION: A high pre-treatment P-VEGF level is auseful marker for tumor nroeression, esBeciallv for vascularinvasion. TACE increases the level of P-VEGF onlytemporarily which may be associated with tumor ischemia.P-VEGF may be useful in predicting treatment response,monitoring disease course after TACE and judging the effectof different TACE regimens.
文摘AIM: To investigate the effect of nutritional support therapy on severe acute pancreatitis (SAP).METHODS: A total of 96 patients with severe acute pancreatitis were divided randomly into control and treatment groups.The former group received total parenteral nutrition (TPN)via central venous infusion, while parenteral nutrition (PN)and enteral nutrition (EN) therapies were applied in different phases for the latter group. The nutrition status, acute phase responses, pancreas lesions, enteric mucosa penetrability and immune functions were monitored.RESULTS: Body weight and prealbumin concentration were increased in treatment group, compared to those in the control group, but albumin concentration did not change significantly.Acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ)scores decreased after 7 d of treatment, whereas the scores of the control group decreased on the 11th day. Concentrations of tumor necrosis factor-α (TNF-α), interleukine-6 (IL-6) and serum C reactive protein (CRP) dropped earlier in the treatment group (on the 4th day) than that in the control group (on the 7th day). No difference was observed in pancreatic lesions between the control and treatment groups.Concentration of endotoxin and lactulose/manicol (L:M) ratio of urine did not change in treatment group, but those in the control group were elevated markedly. Compared with the treatment group, CD4:CD8 T cells ratio and immunoglobulin G (IgG) concentration in the control group decreased significantly.CONCLUSION: Compared to TPN, the combined therapy of EN and PN could improve the nutrition status and moderate the acute phase response obviously. Moreover, the integrity of enteric mucosa and immune function were protected more effectively in treatment group than in the control one. On the other hand, EN did not simulate the excretion of pancreas and avoid exaggerating the inflammation of pancreas. Thus,appropriate application of PN and EN appears to be more effective for patients with SAP.
基金the Science Research Foundation of Schistosomiasis of Hubei Province,No.2000
文摘AIM: To study the effects of pentoxifylline (PTX) on thecontent of hepatic TGF-β1, type Ⅰ and type Ⅲ collagen inschistosomiasis japonica mice with liver fibrosis and itsmechanism of anti-fibrosis.METHODS: Forty mice with schistosomiasis were dividedinto four groups: one group as control without anytreatment, other three were treated with Praziquantel 500mg/(kg.d)for 2 d, high dose PTX 360 mg/(kg.d) for 8 wk,and low dose PTX 180 mg/(kg.d) for 8 wk respectively.Immunohistochemical technique and multimedia colorpathographic analysis system were applied to observe thecontent change of hepatic TGF-β1, type Ⅰ and type Ⅲcollagen in schistosomiasis japonica mice with liver fibrosisbefore and after PTX treatment.RESULTS: Effects of PTX on the content change of hepaticTGF-β1, type Ⅰ and type Ⅲ collagen in schistosomiasis japonicamice with liver fibrosis were related to the dosage of PTX,high dose PTX treated group could significantly reduce thecontent of TGF-β1 (0.709±0.111), type Ⅰ (0.644±0.108) andtype Ⅲ (0.654±0.152) collagen compared with those ofcontrol group (0.883±0.140, 0.771±0.156, 0.822±0.129)with statistical significance (P＜0.05). Low dose PTX couldalso reduce the hepatic content of TGF-β1 (0.752±0.152),type Ⅰ (0.733±0.117) and type Ⅲ (0.788±0.147) collagen,but without statistical significance (P＞0.05). Both high doseand low dose PTX groups have significant differences onthe content of TGF-β1, type Ⅰ and type Ⅲ collagen (P＜0.05,P＜0.05, P＜ 0.01,respectively).CONCLUSION: High dose of PTX treatment could reducethe content of hepatic TGF-β1, type Ⅰ and type Ⅲ collagensignificantly in schistosomiasis japonica mice with liverfibrosis, and thus plays its role of antifibrosis.
基金the Natural Science Foundation of Hubei Province,No.2000J062
文摘AIM: To study the effects of carbon dioxide on the metastatic capability of cancer cells, and to compare them with that of nitrogen.METHODS: The colon cancer cell CCL-228 was treated with 100 % carbon dioxide or nitrogen at different time points and then cultured under normal condition. Twelve hours after the treatment, the survival rates of suspension cells and the expressions of e-cadherin and VEGF were examined.RESULTS: After 60 min of carbon dioxide and longer time of nitrogen treatment, the suspended cells increased and the expression of e-cadherin decreased while the expression of VEGF was enhanced significantly. And the effects of nitrogen were similar to, but weaker than, those of carbon dioxide.CONCLUSION: Carbon dioxide may improve the metastatic capability of cancer cells and its effects are significantly stronger than that of nitrogen. A sequential use of carbon dioxide and nitrogen in pneumoperitoneum may take the advantage of both gases.
基金Supported by the National Natural Science Foundation of China,NO.39970306
文摘AIM:To investigate the effects of autoantibodies against β_1-adrenoceptor in hepatitis virus myocarditis on action potential and L-type Ca^(2+) currents. METHODS:Fifteen samples of autoantibodies against β_1- adrenoceptor positive sera of patients with hepatitis virus myocarditis were obtained and IgGs were purified by octanoic acid extraction.Binding of autoantibodies against β_1- adrenoceptor to guinea pig cardiac myocytes was examined by immunofluorescence.Using the patch clamp technique, the effects on the action potential and I_(ca-L) of guinea pig cardiac myocytes caused by autoantibodies against β_1-adrenoceptor in the absence and presence of metoprolol were investigated. Cell toxicity was examined by observing cell morphology and permeability of cardiac myocytes to trypan blue. RESULTS:The specific binding of autoantibodies against β_1-adrenoceptor to guinea pig cardiomyocytes was observed. Autoantibodies against β_1-adrenoceptor diluted at 1:80 prolonged APO_(20),APD_(50)and APD_(90) by 39.2%,29.1% and 15.2% respectively,and only by 7.2%,5.3% and 4.1% correspondingly in the presence of 1 μmol/L metoprolol. Autoantibodies against β_1-adrenoceptor diluted at 1:80, 1:100 and 1:120 significantly increased the I_(ca-l) peak current amplitude at 0 mV by 55.87±4.39%,46.33±5.01% and 29.29±4.97% in a concentration-dependent manner.In contrast,after blocking of β_1-adrenoceptors (1 μmol/L metoprolol),autoantibodies against β_1-adrenoceptor diluted at 1:80 induced a slight increase of I_(ca-L) peak amplitude only by 6.81±1.61%.A large number of cardiac myocytes exposed to high concentrations of autoantibodies against β_1- adrenoceptor (1:80 and 1:100) were turned into rounded cells highly permeable to trypan blue. CONCLUSION:Autoantibodies against β_1-adrenoceptor may result in arrhythmias and/or impairment of myocardiums in HVM,which would be mediated by the enhancement of I_(ca-L.
文摘AIM:To assess the vascularity of hepatocellular carcinoma (HCC) before and after transcatheter arterial chemoembolization (TACE) with the quantitative parameters obtained by first pass perfusion weighted MR imaging (FP-MRI). METHODS:Seventeen consecutive patients with one to three lesions in liver underwent FP-MRI before treatment. FP-MRI was also performed one,three,six,nine months, and one year after TACE.The baseline signal intensity (SO) of pre-TACE and one month after TACE was analyzed,the vascularity of HCC assessed by steepest slope of the signal intensity versus time curves (SS) was blindly correlated with their DSA feature and clinical outcome. RESULT:No significant difference was found on baseline signal intensity (S0) between pre-TACE and one month after TACE (F=0.309,P=0.583),The SS (mean,32% per second) of lesion one month after TACE was lower than that of pre-TACE (mean,69% per second),but with no statistical significance (F=3.067,P=0.092).When local recurrence occurred,the time intensity curves became steeper.The vascularity of HCC before and after TACE graded by SS closely correlated with that by DSA (K=0.453,P<0.05). CONCLUSION:FP-MRI is a useful criterion for selecting effective interventional treatment for patients with HCC in their initial treatment and during follow up.
基金Supported by Sience and Technology Issue of Fujian Province,No.99-Z-162
文摘AIM: To investigate the effects of hepatitis B virus x gene and its protein product HBxAg on apoptosis in hepatocyte line HL-7702. METHODS: The reconstituted plasmid pcDNA3-x was established through recombination DNA technique; pcDNA3X was transfected into HL-7702 cells by lipid-mediated trasfection. Positive clones were screened by G418, and HL7702/HI3x cells were analysed by the RT-PCR to confirm the steady expression of X gene in HL-7702 cells. The apoptosis rate in HL-7702 cells was determined by flow cytometry, TUNEL technology, electronic microscope. At the mean time, pcDNA3-X was transfected transiently into HL-7702 cells, and total RNA from HL-7702 cells was extracted 24, 48, 72, 96 and 120 h after the transient transfection, and semiquantitative analysis was performed by RT-PCR to detect the expression of HI3V X gene. Furthermore, apoptosis rate in HL-7702 cells was determined by flow cytometry analysis at the different times. RESULTS: RT-PCR analysis showed that HI3V X gene could be expressed stably in HL-7702 cells. Both flow cytometry and TUNEL technology revealed that the apoptosis rates of HL-7702/HI3x cells were much higher than those of HL-7702/ pcDNA3 and HL-7702 cells. Furthermore, the apoptotic phenomena and apoptotic body were observed in HL-7702/ HI3x cells under electronic microscope, but not in HL-7702/ pcDNA3 and HL-7702 cells. In the experiment of transient transfection, RT-PCR reveals that X gene was expressed most at 72 h after transfection; and the apoptosis rate reached the highest at the same time. After that, the apoptosis rate was reduced with the decrease of the X gene expression. CONCLUSION: HBV X gene and X protein can promote the apoptosis in hepatocyte. And there exist a quantity-effect relationship between the X gene expression and apoptosis rate in hepatocyte.
文摘AIM: To observe the change of tumor microcirculation after transcatheter arterial chemoembolization (TACE) with bletilla microspheres by using first pass perfusion MR imaging (FP) and Chinese ink casting.METHODS: VX2 carcinoma cells were surgically implanted into the left and right lobes of liver of 30 New Zealand white rabbits, which were divided into 3 groups at random. Emulsion of lipiodol mixed with mitomydn C, and 5-FU bletilla microspheres were injected into the hepatic artery respectively, and saline was used as control agent. MR imaging was performed with turbo-flash sequence 14 d after tumor implantation and 7 d after interventional therapy. The steepest slopes (SS) of the signal intensity versus time curves were created for quantitative analysis, 7.5% Chinese ink gelatin solution was injected through ascending artery (17 cases) or portal vein(2 cases) for lesion microvessel area (MVA) measurement after the last MRI examination.The correlation between perfusion imaging and MVA was studied blindly.RESULTS: The SS values at the rim of tumor in lipiodol group (mean, 49% per second) and bletilla group (mean,35% per second) were significantly decreased (P<0.05) as compared with control group (mean, 124% per second), no difference was found between lipiodol and bletilla groups(P>0.05). In lipiodol group, the MVAs (24 974±11 836μm^2) in the center of the tumor were significantly smaller than those of the control group (35 510±15 675 μm^2) (P<0.05),while the MVAs (80 031±22 745 μm^2) around the tumor were significantly increased because small and dense plexuses appeared around the tumor which correlated to intense reaction of granulation tissue. None of the vessels was seen in the tumor in bletilla group, the peripheral MVAs of the tumor were significantly smaller than those of the control group (P<0.05) and lipiodol group (P<0.05). There was a good correlation between SS and MVAs in control group (rsl, 0.985, P<0.0001) and bletilla group (rsl, 0.743,P<0.05), the correlation was not significant in lipiodol group(rsl, 0.527, P>0.05).CONCLUSION: TACE with bletilla microspheres may enhance its anti-tumor effect by inhibiting the angiogenesis,and FP-MRI provides useful information to assess the TACE effect by depicting tumor vascularization and perfusion,
基金Supported by the National Natural Science Foundation of China,No.39770839
文摘AIM:Transcatheter arterial embolization (TAE) of the hepatic artery has been accepted as an effective treatment for unresectable hepatocellular carcinoma (HCC). However,embolized vessel recanalization and collateral circulation formation are the main factors of HCC growth and recurrence and metastasis alter TAE. Vascular endothelial growth factor (VEGF) plays an important role in tumor angiogenesis.This study was to explore the inhibitory effect of VEGF antisense oligodeoxynucleotides (ODNs) on VEGF expression in cultured Walker-256 cells and to observe the anti-tumor effect of intra-arterial infusion of antisense ODNs mixed with lipiodol on rat liver cancer.METHODS: VEGF antisense ODNs and sense ODNs were added to the media of non-serum cultured Walker-256 cells.Forty-eight hours later, VEGF concentrations of supernatants were detected by EUSA. Endothelial cell line ECV-304 cells were cultured in the supernatants. Seventy-two hours later,growth of ECV-304 cells was analyzed by NTT method. Thirty Walker-256 cell implanted rat liver tumor models were divided into 3 groups.0.2 mL lipiodol (LP group, n=10), 3OD antisense ODNs mixed with 0.2 mL lipiodol (LP+ODNs group, n=10) and 0.2 mL normal saline (control group, n=10) were infused into the hepatic artery. Volumes of tumors were measured by MRI before and 7 d alter the treatment.VEGF mRNA in cancerous and peri-cancerous tissues was detected by RT-PCR. Microvessel density (MVD) and VEGF expression were observed by immunohistochemistry.RESULTS: Antisense ODNs inhibited Walker-256 cells' VEGF expression, The tumor growth rate was significantly lower in LP+ODNs group than that in LP and control groups (140.1±33.8%, 177.9±64.9% and 403.9±69.4% respectively, F=60.019,P<0.01).VEGF mRNAs in cancerous and peri-cancerous tissues were expressed highest in LP group and lowest in LP+ODNs group. The VEGF positive rates showed no significant difference among LP, control and LP+ODNs groups (90%,70% and 50%, H=3.731, P>0.05).The MVD in LP+ODNs group (53.1±18.4) was significantly less than that in control group (73.2±20.4) and LP group(80.3±18.5) (F=5.44, P<0.05).CONCLUSION: VEGF antisense ODNs can inhibit VEGF expression of Walker-256 cells.It may be an antiangiogenesis therapy agent for malignant tumors. VEGF antisense ODNs mixed with lipiodol embolizing liver cancer is better in inhibiting liver cancer growth, VEGF expression and microvessel density than lipiodol alone.
基金Supported by the National Natural ScienceFoundation of China,No.30240022
文摘AIM: TO investigate the effects of anti-Fas ribozyme on Fas expression and apoptosis in primary cultured mouse hepatocytes.METHODS: Mouse hepatocytes were isolated by using collagenase irrigation. A hammerhead ribozyme targeting the Fas mRNA was constructed, and transfected into mouse hepatocytes via Effectene. Then Fas expression in mouse hepatocytes was detected by RT-PCR and western blotting.After being treated with anti-Fas antibody (JO2), hepatocytes viability was measured with MTI- assay. Caspase-3 proteolytic activity was detected, and cell apoptosis was measured according to Annexin V-FITC apoptosis detection kit.RESULTS: Fas expressed in primary mouse hepatocytes.Fas expression in hepatocytes transfected with anti-Fas ribozyme was decreased remarkably and correlated with the resistance to Fas-mediated apoptosis as determined by flow cytometry and caspase-3 proteolytic activity.CONCLUSION: AnU-Fas ribozyme can remarkably decrease the Fas expression in mouse hepatocytes, thus inhibit Fasmediated apoptosis in hepatocytes, It is suggested that anti-Fas ribozyme could significantly increase the resistance of transplanted hepatocytes to apoptosis and improve the survival of transplanted hepatocytes,
基金the Natural Science Foundation of Hubei Province, No.2000J042the Science Research Foundation of the Education Office of Hubei Province,No.2000A06010
文摘AIM: To investigate the effect of Kangxian ruangan keli (KXR) on hepatic stellate cell (HSC) proliferation mediated by platelet-derived growth factor (PDGF) and the underlying mechanism.METHODS: In a serum-free culture system, HSCs were treated with a KXR preparation for 24 hours, followed by stimulation with PDGF-BB for 24 hours. Then the cells were incubated again in the medium containing KXR for 3 hours stimulated with PDGF-BB for 5 minutes, and collected. The proliferation of HSC was examined using an MTT assay and flow cytometry. Tyrosine phosphorylation was detected with Western blotting and visualized by the enhenced chemiluminescent (ECL) method.RESULTS: The OD values for the HSCs growing in the media without and with addition of PDGF were 0.17±0.06 and0.82±0.05, respectively. The PDGF-induced increase was hindered remarkably by KXR preparation in a dose-dependent manner. The reaction values for the systems with 5mg/mL, 2.5 mg/mL and 1.25 mg/mL of KXR were 0.28±0.03,0.37±0.02 and 0.43±0.04, respectively. Moreover, the percentages of S-phase cells in these KXR-containing culturesystems were 10.95±1.35, 32.76±1.07 and 43.19±1.09,respectively, all of which were significantly lower than that in the culture free of KXR (68.24±2.72). In addition, the values for tyrosine-phosphorylated protein in HSCs treated with 5 mg/mL and 1.25 mg/mL of KXR were 0.1349±0.0072 and 0.1658±0.0025, respectively, which were smaller than that in the cells treated only with PDGF-BB (0.1813±0.0117).CONCLUSION: Within the dose range used in the present study, KXR preparation shows an inhibitory effect on HSC proliferation induced by PDGF. The mechanism of this process may involve interference with tyrosine phosphorylation mediated by PDGF.
基金National Natural Science Foundation of China, No 30200273 & 30672067
文摘AIM： To investigate the role of SMYD3 in hepatocellular carcinoma （HCC） development and progression and to verify whether its regulation activity was through RIZ1 inactivation. METHODS： Expression of SMYD3 in HCC cell lines and tissues were measured; silencing of SMYD3 by RNA interference （RNAi） was effectuated, hepatoma cell proliferation, migration and apoptosis were tested, with RIZl CpG promoter methylation, and corresponding mRNA expression were investigated. RESULTS： SMYD3 over-expression in HCC was associated with RIZl hypermethylation and mRNA down-expression. Suppression of SMYD3 expression de- methylated RIZl CpG promoter （P 〈 0.01） and increased RIZl mRNA expression （P 〈 0.01）. Consequently, SMYD3 down-expression with RIZl de-methylation strongly inhibited hepatoma cell growth （MTT inhibitory rates： Pgenesil-1-s1 60.95%± 7.97%, Pgenesil-1-s2 72.14% ± 9.68% vs Pgenesil-1-hk 6.89% ± 4.12%, P 〈 0.01） and migration （Pgenesil-1-s1 4.24% ± 1.58%, Pgenesil- 1-s1 4.87% ± 0.73% vs Pgenesil-1 19.03% ± 4.63%, Pgenesil-1-hk 19.95% ±5.21%, P 〈 0.01） and induced apoptosis （FCM subG1 phase Pgenesil-1-s1 19.07% ＋ 1.78%, Pgenesil-1-s2 17.68% ± 2.36% vs Pgenesil-1 0.47% ± 0.12%, Pgenesil-1-hk 1.46% ± 0.28%, P 〈 0.01. TUNEL-positive cells： Pgenesil-1-s1 40.24%± 5.18%, Pgenesil-1-s2 38.48% ± 4.65% vs Pgenesil-1 2.1B% - 1.34%, Pgenesil-1-hk 2.84%± 1.22%, P 〈 0.01） in HepG2 cells. CONCLUSION： These results demonstrate that SMYD3plays a critical role in the carcinogenesis and progression of HCC, The proliferation, migration induction and apoptosis inhibition activities of SMYD3 may be mediated through RIZl CpG promoter hypermethylation.
文摘AIM: To study the distribution and stability of antisense oligodeoxynucleotide (ASODN) in Walker-256 cells and their distribution in liver, lung and kidney tissues after being infused alone or mixed with lipiodol via hepatic artery in a rat liver tumor model.METHODS: 5'-Isothiocyananate (FITC)-labeled vascular endothelial growth factor (VEGF) ASODN was added into Walker-256 cell culture media. Its distribution in cells was observed by fluorescence microscope at different time points. Walker-256 carcinosarcoma was transplanted into Wistar rat liver to establish a liver cancer model. 5'-FITC-labeled VEGF ASODN mixed with (mixed group, n = 6) or without (TAI group, n = 6)ultra-fluid lipiodol was administrated via hepatic artery.Frozen samples of liver, lung and kidney tissue were taken from rats after 1, 3 and 6 d, respectively. The distribution of ASODN was observed under fluorescent microscope.RESULTS: ASODN could enter cytoplasm within 2 h and nuclei within 6 h. Accumulation of ASODN reached the peak point in nuclei at 12 h, and then disappeared gradually. No fluorescence could be seen in cells at 48 h. In vivo experiment, on d 1 and 3 the fluorescence staining in liver was stronger in mixed group than in TAI group and more fluorescence could be detected in lung and kidney in TAI group than in mixed group. On d 6, no fluorescence could be detected in TAI group, but faint fluorescence could be seen in mixed group. ASODN could be seen in cancer cells and normal hepatic cells. In mixed group, ASODN was mainly distributed in liver tumor tissues.CONCLUSION: ASODN can transfect Walker-256 cells.ASODN mixed with lipiodol infusion via hepatic artery can be used in the treatment of HCC.
文摘AIM: This study was designed to examine the hypothesis that gender differences in I/R injury are associated withendothelial cell nitric oxide synthase (eNOS)-derived nitric oxide (NO).METHODS: Wistar rats were randomized into seven experimental groups (12 animals per group). Except for the sham operated groups, all rats were subjected to total liver ischemia for 40 min followed by reperfusion. All experimental groups received different treatments 45 min before the laparotomy. For each group, half of the animals (six) were used to investigate the survival; blood samples and liver tissues were obtained in the remaining six animals after 3 h of reperfusion to assess serum NO, alanine aminotransferase (ALT) and TNF-α levels, liver tissuemalondialdehyde (MDA) content, and severity of hepatic I/R injury.RESULTS: Basal serum NO levels in female sham operated (FS) group were nearly 1.5-fold of male sham operated (MS) group (66.7±11.0 μmol/L vs 45.3±10.1μmol/L, P＜0.01). Although serum NO levels decreased significantly after hepatic I/R (P＜0.01, vs sham operated groups), they were still significantly higher in female rat (F) group than in male rat (M) group (47.8±8.6 μmol/L vs 23.8±4.7 μmol/L, P＜0.01). Serum ALT and TNF-α levels, and liver tissue MDA content were significantly lower in F group than in M group (370.5±46.4 U/L, 0.99±0.11 μg/L and 0.57±0.10 μmol/g vs668.7±78.7 U/L, 1.71±0.18 μg/Land 0.86±0.11 μmol/g, respectively, P＜0.01). I/R induced significant injury to the liver both in M and F groups (P＜0.01 vs sham operated groups). But the degree of hepatocyte injury was significantly milder in F group than in M group (P＜0.05 and P＜0.01). The median survival time was six days in F group and one day in M group. The overall survival rate was significantly higher in F group than in M group (P＜0.05). When compared with male rats pretreated with saline (M group), pretreatment of male rats with 17-β- estradiol (E2) (M+E2 group) significantly increased serum NO levels and significantly decreased serum ALT and TNF-α levels, and liver tissue MDA content after I/R (P＜0.01).The degree of hepatocyte injury was significantly decreased and the overall survival rate was significantly improved in M+E2 group than in M group (P＜0.01 and P＜0.05). TheNOS inhibitor Nw-nitro-L-arginine methyl ester (L-NAME) treatment could completely abolish the protective effects of estrogen in both male and female rats. CONCLUSION: The protective effects afforded to female rats subjected to hepatic I/R are associated with eNOSderived NO.
基金Supported by the-Foundation of Education Committee of Fujian Province,No.K99054
文摘AIM: To explore the cooperative effects of antisense oligonucleotide (ASON) of cell adhesion molecules and cimetidine on the expression of E-selectin and ICAM-1 in endothelial cells and their adhesion to tumor cells. METHODS: After treatment of endothelial cells with ASON and/or cimetidine and induction with TNF-α, the protein and mRNA changes of E-selectin and ICAM-1 in endothelial cells were examined by flow cytometry and RT-PCR,respectively. The adhesion rates of endothelial cells to tumor cells were measured by cell adhesion experiment. RESULTS: In comparison with TNF-α inducing group, lipoASON and lipo-ASON/cimetidine could significantly decrease the protein and mRNA levels of E-selectin and ICAM-1 in endothelial cells, and lipo-ASON/cimetidine had most significant inhibitory effect on E-selectin expression (from 36.37±1.56% to 14.23±1.07%, P<0.001). Meanwhile,cimetidine alone could inhibit the expression of E-selectin(36.37±1.56% vs 27.2±1.31%, P<0.001), but not ICAM-1(69.34±2.50% vs68.07±2.10%,P>0.05)and the two kinds ofmRNA, either. Compared with TNF-α inducing group, the rate of adhesion was markedly decreased in lipo-E-selectin ASON and lipo-E-selectin ASON/cimetidine treated groups(P<0.05),and lipo-E-selectin ASON/cimetidine worked better than lipo-E-selectin ASON alone except for HepG2/ECV304 group(P<0.05). However, the decrease of adhesion was not significant in lipo-ICAM-1 ASON and lipo-ICAM-1 ASON/dmetidine treated groups except for HepG2/ECV304 group (P >0.05). CONCLUSION: These data demonstrate that ASON in combination with cimetidine in vitro can significantly reduce the adhesion between endc^thelial cells and hepatic or colorectal cancer cells, which is stronger than ASON or cimetidine alone. This study provides some useful proofs for gene therapy of antiadhesion.
文摘To determine the accuracy of triphase enhanced helical CT in judging the invasion and metastasis of gastric carcinoma, and to discuss the relation between imaging signs and pathological findings. Methods: Triphase enhanced helical CT scanning was performed in 46 patients with gastric carcinoma. Imaging findings were compared with postoperative pathologic results. Results: (1) The accuracy of helical CT for diagnosing involvement of tunica serosa, lymph node metastasis and distant metastasis was 84.8%, 87.0% and 100~ respectively. (2) CT signs of serosal involvement, lymph node metastasis and distant metastasis were in good accordance with pathological findings (P<0.05). Conclusion: Triphase enhanced helical CT scans can comprehensively and precisely reflect the pathologic characteristics of gastric carcinoma, thus it is a reliable technique for the diagnosis of this disease.
文摘AIM: To study the effects of palmatine, a known inhibitoron delayed rectifier potassium current and L-type calciumcurrent (ICa,L) in guinea pig ventricular myocytes, on thepotassium and calcium currents in isolated rat hepatocytes.METHODS: Tight-seal wh ole-cell patch-clamp techniqueswere performed to investigate the effects of palmatine onthe delayed outward potassium currents (IK), inward rectifierpotassium current (IK1) and Ca2+ release-activated Ca2+current (ICRAC) in enzymatically isolated rat hepatocytes.RESULTS: Palmatine 0.3-100 μM reduced IK in a concentationdependent manner with EC50of 41.62±10.11 μM and nH,0.48±0.07 (n=8). The effect of the drug was poorly reversibleafter washout. When the bath solution was changed totetraethylammonium (TEA) 8 mM, IK was inhibited.Palmatine 10 μM and 100 μM shifted the I-V curves of IKdownward, and the block of IK was voltage-independent.Palmatine 0.3-100 μM also inhibited ICRAC in a concentration-dependent manner. The fitting parameters were as follows:ECs0=51.19±15.18 μM, and nH=0.46+0.07 (n=8). The peakvalue of ICRAC in the I-V relationship was decreased bypalmatine 10 μM and 100 μM. But the reverse potential ofIcRAcoCcurred at Voltage=0 mV in all cells. Palmatine 0.3-100 μM failed to have any significant effect on either inwardor outward components of IK1 at any membrane potentialexamined.CONCLUSION: The inhibitory effects on IK and ICRAC couldbe one of the mechanisms that palmatine exerts protectiveeffect on hepatocytes.
文摘AIM: To examine the effects of berberine, an isoquinoline alkaloid with a long history used as a tonic remedy for liver and heart, on ion channels of isolated rat hepatocytes.METHODS: Tight-seal whole-cell patch-clamp techniques were performed to investigate the effects of berberine on the delayed outward potassium currents (Ik), inward rectifier potassium currents (Ik1) and Ca^2+ release-activated Ca^+ currents (ICRAC) in enzymatically isolated rat hepatocytes.RESULTS: Berbenne 1-300 μmol/L reduced/K in a concentration-dependent manner with EC50 of 38.86=1=5.37 μmol/L and nH of 0.82±0.05 (n = 8). When the bath solution was changed to tetraethylammonium (TEA) 8 retool/L,IK was inhibited.Berberine 30 μmol/L reduced/K at all examined membranepote ntials, especially at potentials positive to +60 mV (n = 8,P<0.05 or P<0.01 vs control). Berberine had mild inhibitory effects on IK1 in rat hepatocytes. Berberine 1-300 μmol/L also inhibited ICRAC in a concentration-dependent fashion. The fitting parameters were EC50 = 47.20±10.86 μmol/L,nH= 0.71±0.09 (n = 8). The peak value of/CRAC in the I-Vrelationship was decreased by berberine 30 μmol/L at potentialnegative to -80 mV (n = 8, P<0.05 vscontrol). But the reverse potential of/CRAC occurred at voltage 0 mV in all cells.CONCLUSION: Berberine has inhibitory effects on potassium and calcium currents in isolated rat hepatocytes, which may be involved in hepatoprotection.
文摘To test the accuracy of real-time three-dimensional echocardiography (RT3DE) imaging system for evaluating left ventricular mass (LVM) in phantom and excised canine heart. Methods Ten left ventricular (LV) wall phantoms made of two rubber-bursas, ten excised canine hearts underwent RT3DE and two-dimensional echocardiography (2DE). In RT3DE "full volume" imaging, the myocardial volume was mea-sured using 2,4,8, and 16-plane method with the analysis software of RT3DE. Mass was then calculated by multiplying the resulting myocardial volume by specific density of myocardial tissue. In 2DE the masses were measured by area-length meth-od. The true LV wall phantom mass was measured by water displacement and the canine LVM was weighed by anatomy, which served as a reference standard. We compared RT3DE or 2DE with true mass. Results In LV wall phantoms, RT3DE correlated with true masses strongly (r = 0.813-0.994) and weakly correlated between 2DE and true masses (r = 0.628). In excised canine hearts, there is an excellent correlation between RT3DE and true masses (r = 0.764-0.991), while 2DE value showed a lesser correlation (r = 0.514). There are no difference between RT-3DE and true masses (P > 0.05) but different between 2DE and true masses (P < 0.05). In different planes, there was no difference between 8-plane and 16-plane (P > 0.05) but different between 8-plane and 2, 4-plane (P < 0.05). Conclusion RT3DE can accurately quantify LVM and provide a new tool to evaluate LV function. For LVM by RT3DE, 8-plane measurement method is the best choice for accuracy and convenience.
文摘We report two cases of extrahepatic portal vein aneurysm,and both of them underwent surgical intervention. The first case had a mild pain in right upper quadrant of the abdomen; the second had no obvious symptoms. Physical examination revealed nothing abnormal. Both of them were diagnosed by magnetic resonance imaging angiography (MRA). One of the aneurysms was located at the main portal vein, the other, at the confluence of the superior mesenteric vein and the splenic vein, and these two places are exactly the most common locations of the extrahepatic portal vein aneurysm reported in the literature (30.7% each site). The first case underwent aneurysmorrhaphy and the second case, aneurysm resection with splenectomy. Both of them recovered soon after the operation, and the symptom of the first case was greatly alleviated. During the follow-up of half a year, no complication and adverse effect of surgical intervention was found and the color Doppler ultrasonography revealed no recurrence of the aneurysmal dilation. We suggest that surgical intervention can alleviate the symptom of the extrahepatic portal vein aneurysm and prevent its complications effectively and safely for low risk patients.
