Objective To establish a stable transplanted model of Vx 2 carcinoma in rabbit from three different transplanted methods, and analyze the DSA (digital substraction angiography) features of the implanted tumors. Methods 60 white rabbits of New Zealand were divided into 3 groups at random, 20 for each group. In the first group, Vx 2 carcinoma cells (about 5?10 7 )were infused via hepatic artery into the rabbit liver. For the second group, Vx 2 carcinoma cells (about 5?10 7 ) were inplanted into the left lobe of the liver via laparotomic route and in the 3rd group with the same route as the 2nd group, a piece of tumor tissue (contained about 10 6 10 8 carcinoma cells) were implanted into the left lobe of the liver. And then a comparative observation was performed in 3 groups included:1. the successful rate of implanted tumors; 2. the volume change and the growth rate of implanted tumors; 3. the biological features of implanted tumors in histopathology and electronic microscope photographs and DSA features of implanted tumors. Results The successful rates of transplanted tumors in the three groups were 7/20, 10/20 and 19/20 respectively. The successful rate of the 3rd group was the highest of the 3( P

Objectives:To explore the inhibition effect of TK/GCV and CD/5-FC system on liver metastasis of colon carcinoma.Methods:Forty nude mice were divided into 4 groups randomly,which were the control group,TK group,CD group and TK-CD group with 10 nude mice in each group.The mice in the control group were injected SW480 cells in the spleen and NS into the peritoneal cavity.The mice in other groups were all injected SW480/TK-CD cells in the spleen.And GVC,5-Fc,GVC+5F-c were injected into the peritoneal cavity of the mice in the TK group,CD group and TK-CD group respectively.Liver metastasis rate,the number of liver metastasis,the pathological changes of the tumor tissues under the electronic microscope,tumor cell's apoptotic index and life span and other indexes of nude mice in four groups were analyzed.Results:Liver metastasis rate of nude mice in every treatment group was lower than that in control group.Compared with control group,average number of liver metastasis decreased,life span of nude mice was prolonged,and the apoptosis rate of cancer cell in liver metastasis increased.The combined genes in the TK-CD group worked more markedly,and there was mutual effect between TK/GCV and CD/5-Fc.Conclusions:Synergistic effect was acquired combining TK/GCV with CD/5-FC system and it inhibited the formation of liver metastasis of colon carcinoma efficiently.

Objective To demonstrate the inhibitory effect of adenosine A1 receptor agonist R(-)-N6-(2-phenylisopropy1)adenosine(R-PIA)and cross-talk between adenosine A1 receptor and CaN in cultured myocardial cells hypertrophy induced by Ang Ⅱ.Methods The protein synthetic rate was determined by measuring the incorporation of [3H]-leucine into myocyte protein;the cardiomyocytes volumes were measured by computer photograph analysis system;the expression of atrial natriuretic polypeptide(ANP)and CaN were determined by Western-blot.Results R-PIA(1 ?mol/L)inhibited Ang Ⅱ(0.1 ?mol/L)-induced increase of [3H]-leucine incorporation [R-PIA:(976.2?89.0)vs Ang Ⅱ:(1130.7?102.7)count per min per well,P

Objective To investigate the changes in serum levels of nitric oxide ( NO) and endothelin ( ET) in type 2 diabetes patients with vascular complications, and to analyze the relationship between these levels and risk factors.Method We selected 98 cases of type 2 diabetes patients.Based on the grouping criteria, the patients were divided into diabetic patients with vascular complications ( group A,49 cases) and those without ( group B,49 cases) .In addition, 44 age-and body mass index-matched healthy cases were selected for control(group C).Height, weight, blood pressure, duration of diabetes, fasting blood glucose, hemoglobin A1c ( HbA1c), blood lipids, and serum levels of NO and ET-1 of all the patients were recorded.Results The NO levels of the two groups with diabetes mellitus were significantly lower than in group C[(43.87 ±12.05)and (53.29 ±11.75)μmol/L versus (66.08 ±16.48)μmol/L, P<0.01], while the ET-1 levels of the two groups with diabetes mellitus were significantly higher [(100.25 ±20.34) and (77.55 ±14.84) versus (53.62 ±8.40)ng/L, P<0.01] than those of the group C.The NO levels of group A were significantly lower than in group B [(43.87 ±12.05) versus (53.29 ±11.75)μmol/L, P<0.01].Moreover, the ET-1 levels of the group A were significantly higher than in group B [(100.25 ±20.34) versus (77.55 ±14.84)ng/L, P<0.01].Between the two diabetic groups, group A showed significantly higher systolic blood pressure(SBP), diastolic blood pressure(DBP), HbA1c, and course than group B (P<0.01).Correlation analysis showed a negative correlation between SBP, DBP, fasting blood glucose, HbA1c, and NO a positive correlation between high-density lipoprotein cholesterol ( HDL-C ) and NO, a negative correlation between HDL-C and ET-1, and a positive correlation between SBP,LDL-C, uric acid, fasting blood glucose, HbA1c, and ET-1.Conclusion The serum levels of NO and ET-1 in diabetic patients are evidently abnormal.Vascular endothelium injury will become more serious in patients with complications.Therefore, the serum levels of NO and ET-1 in diabetic patients are correlated with control of blood glucose, blood pressure, and blood lipid levels.

AIM: To compare the impacts of adriamycin(ADM) intermittent and continuous heat infusion on respiration, heart rate and temperature of the rabbit as well as the concentration of ADM in VX-2 tumor for the verification of the safety and effectiveness of intermittent heat infusion through VX-2 transplanted tumor model in rabbit liver.METHODS: VX-2 tumor models were established respectively in the thighs of 30 New Zealand rabbits. All 30 rabbits were randomly allocated into three groups with 10 rabbits each. After catheterizing into femoral artery, which was demonstrated as the supply artery of tumors by digital subtraction angiography(DSA), animals in three groups were infused with 100 mL saline at normal temperature and ADM, 100 mL saline at 60 ℃ and ADM continuously and 100 mL saline at 60 ℃ and ADM intermittently respectively. During infusion, the 43 ℃ to 45 ℃ lasting time in the tumor tissues of the two 60 ℃ groups was measured. After infusion, the respiration(time/minute), heart rate(beats/minute) and temperature(℃ )of the rabbits in each group as well as the concentrations of ADM in tumor tissue were measured immediately.RESULTS: The concentration of ADM was (7. 1 ± 2.2) mg/L in normal temperature infusion group, (17.2 ± 1.6) mg/L in 60 ℃ continuous infusion group and(16.5 ±3.4) in 60 ℃ intermittent infusion group. There were significant differences among three groups( F = 48.95, P = 0. 000), but there was no significant difference between 60 ℃ intermittent infusion group and 60 ℃ continuous infusion group( P ＞ 0. 05). 43℃ to 45 ℃ lasting time was(22.5 ±1.4) minutes in 60 ℃continuous infusion and (24.3±2.4)minutes in 60 ℃intermittent infusion group( F =4.20, P ＞ 0.05) . There were significant differences among three groups in respiration, heart rate and temperature( F = 14. 58, 33.07, 10.00, P ＜ 0.01), but there was no significant difference in the respiration, heart rate and temperature between 60℃ intermittent infusion and normal temperature infusion group ( P ＞ 0. 05).CONCLUSION: Intermittent thermochemotherapy is a more effective and safe interventional thermochemotherapy method compared with continuous heat infusion.

BACKGROUND: There is an intimate temporal and spatial relationship between growth of primitive cardiac cells, septum transversum mesenchyme and liver development. The signal from primitive cardiac cells and septum transversum mesenchyme induces the ventral foregut endoderm cells specialize toward hepatocytes. While the septum transversum mesenchyme provides a suitable environment for forming the liver bud and promoting the growth and differentiation. However, the molecular mechanism of this induction is not yet delineated.OBJECTIVE: Using alpha-fetal protein (AFP), c-Met and cytokeratin (CK) 19 as markers of hepatic stem cells, the growth of early human embryo of 3-5 weeks and morphologic characteristic of hepatic stem cells were observed to demonstrate the characteristic and factors that affected the proliferation and differentiation of hepatic stem cell, which provided experimental evidence for basic research and clinical application of hepatic stem cells.DESIGN: An opening experiment was designed.SETTING: Department of Anatomy, Weifang Medical College.MATERIALS: The experiment was carried out at the Scientific Research Center of Chengdu Medical College between September 2004 and January 2005. Twenty cases fresh human embryos aged less than 2 months were collected with signed agreements of the pregnant women suffering from pregnancy termination with mifepristone. The samples were fixed with 40 g/L polymerisatum in 20 minutes and embedded routinely in paraffin, and then 5 μm thick series sections were continuously made. After hematoxylin-eosin staining, the embryonicage was determined under the microscope according to the length of embryos, the number of somites and the development of organs, which was referring to the Jirasek's staging standard of human embryo.METHODS: The immunohistochemical staining was conducted with SABC method on one of every ten sections, which were incubated overnight at 4 ℃ with polyclonal antibodies against hepatocyte growth factor (HGF),c-Met, insulin-like growth factor (IGF-Ⅰ), IGF-Ⅰ receptor (IGF-IR), transforming growth factor (TGFβ1), TGFβR1, TGFβR2 or monoclonal antibodies against proliferating cell nuclear antigen (PCNA), AFP and CK19.The following day, the sections were incubated for 2 hours at room temperature with biotinylated anti-mouse or anti-rabbit IgG and SABC liquid respectively, and then diaminobenzidine (DAB) was used to color them. The negative control was conducted with the phosphate buffer, then the sections were observed and photographed under light microscope.MAIN OUTCOME MEASUERS: ①the morphologic characteristic of human hepatic stem cells and immunohistochemical staining of markers②the expression of HGF, IGF-Ⅰ, TGFβ1 and their receptors on human embryonic livers of 3-5 weeks, primitive cardiac cells and septum transversum mesenchyme.RESULTS: ①The morphologic characteristic of human hepatic stem cells and immunohistochemical staining of markers: The hepatic bud formed at the end of 3rd week and migrated into the septum transversum mesenchyme to form the hepatic cords at the 4th week. The cells structuring the hepatic cords displayed the typical characteristic of immature cells. At the 5th week, the number of cells within the hepatic cords, the size of cell body,the cytoplasmic acidophilia all increased, whereas the basophilia of nuclei decreased. However the cellular forms were still homogeneous and displayed the typical characteristic of immature cells. The cells of hepatic cords were negative for PCNA response during 3rd-4th week but began to express positive at the 5th week, mainly in the nucleus and minority cellular cytoplasm showed weak positive. Most hepatic cells during 3rd-5th weeks were positive for AFP, c-Met and negative for CK19. The immunologic reaction depositors of AFP positive cells were located in the nuclei, cytoplasm and membrane of the hepatocytes, and c-Met presented mainly in the nuclei and the positive signal was weak in the cytoplasm. ②Expressions of HGF, IGF-Ⅰ, TGFβ1 and their receptors in the embryonic human liver, primitive heart and septum transversum mesenchyme: At the 4th week,the c-Met expressed only in all hepatocytes, whereas the other growth factors and their receptors were undetectable. At the 5th week, all the growth factors, except HGF, were expressed in the hepatocytes. The immunologic reaction depositors of TGFβ1, TGFβ1R1 and TGFβ1R2 were located in the cytoplasm and cell membrane. The positive response of IGF-Ⅰ and IGF-IR were present at nuclei, cytoplasm and cell membrane. At the 3rd-5th week, myocardial cells surrounding liver bud or hepatic cord and the septum transversum mesenchyme were positive for HGF, TGFβ1 and IGF-Ⅰ,with the signals were aggregated mainly in cytoplasm and minority nucei.CONCLUSION: ①It was at the end of 3rd week that part of endoderm cells in foregut ventral were specialized to hepatic stem cells. ②The undifferentiated hepatic stem cells could be drawn to develop to the liver stem cells with bi-directional differentiation potentials by using specific markers for studying human embryonic liver stem cells. According to the corresponding relation of embryonic age between human and rats, the time studying the rat hepatic stem cells could be calculated. ③HGF, IGF-Ⅰ,TGFβ1 and their receptors promoted the early development of human embryonic liver.

Objective To evaluate the effects of interventional thermotherapy(60℃)via hepatic artery on the vascular permeability of hepatic tumor tissue and normal liver tissue in VX2 tumor-bearing rabbits.Methods Thirty white rabbits were used in this experiment,Thirty VX2 tumor-bearing rabbits models were established by direct injection of VX2 tissue particle into the liver parenchyma and were randomly and equally divided into three groups:(1)non-perfusion group(n=10),used as control group; (2)normal thermal perfusion group(n=10),a perfusion of 30ml saline at 37 4±0.5℃ via hepatic artery in 15 minutes was used;(3)hyper-thermai perfusion group(n=10),a perfusion of 30ml saline at 60±0.5℃via hepatic artery in 15 minutes was used.The vascular permeability of the tumor tissue and the normal liver tissue was estimated with Evan's blue method.Results In all three groups the vascular permeability was significantly incleased in tumor tissue than that in normal liver tissue(P＜0.05).Normal thermal perfusion resulted in a significant increase in the permeability of vasculature(JP＞0.05).Hyper-thermal peffusion resulted in a significant increase in vascular permeability of tumor tissue (P＜0.05),and vascular permeability was increased in tumor tissue mole markedly than that in normal tissue(P＜0.05).Conclusion Hyperthermia hepatic arterial perfusion can increase vascular permeability for both tumor tissue and normal tissue in VX2 tumor-bearing rabbits.

Objective To establish a rabbit VX2 liver tumor model by transplanting tumor tissue mass into the rabbit's liver,to analyze and observe the growing features of the liver tumor.Methods The tumor tissue mass(about 106-108 VX2 liver tumor cells)was inoculated into the left hepatic labe in 20 rabbits to establish rabbit VX-2 hepatic carcinoma model.The observation included the following two respects.(1)The tumor's volume at 7,10,14,17 and 21 days after the procedure was measured by ultrasonography and the growth rate of tumor was calculated.(2)The morphological feature of the tumor was inspected both macroscopically and microscopically.Results The growing pattern of the tumor was compatible with the exponential curve.Seventeen days after transplantation the increase rate of the tumor volume was much higher than that of the tumor diameter. Histopathologjcally,the growing pattern of the tumor took the form of infiltrative way,with its appearance being quite similar to the VX2 squamous cell carcinoma.Conclusion Transplantation of tumor tissue mass is the technique of first choice to establish the VX2 liver carcinoma model in rabbits.This experimental model is a very ideal animal form for both clinical and fundamental studies of liver carcinoma.

Objective To study the changes in the immune function of advanced hepatic carcinoma patients after thermochemoembo-lization via hepatic artery. Methods Forty advanced hepatic carcinoma patients were randomized divided into 2 groups. A group (n=20) was treated by perfusion with ADM(40mg) + MMC(10mg) in 37℃ 9% sodium chloride solution via hepatic artery, and B group (n=20) was treated by perfusion with ADM(40mg) + MMC(10mg) in 60℃ 9% sodium chloride solution via hepatic artery, then all were embolized with some quantity of lipiodol. Flow cytometry (FCM) was used to detect the changes in percentages of T-lymphocyte subsets (CD3<'3, CD4<'+, CD8<'+) and NK cells, and the contents of soluble interleukin-2 receptor (sIL-2R) were detected by double sandwich ELISA before and after therapy. LDH enzyme-release assay was used to detect cytotoxic activity of NK cells. Results Compared with pre-therapy group or A group after therapy, the percentage of CD4<'+ cells of B group after therapy significantly increased (P<0.05), the percentages of CD8<'+ cells and the content of sIL-2R of B group after therapy markedly decreased (P<0.05). The percentages of NK cells and cytotoxic activity NK cells of B group after therapy were significantly higher than those from pre-therapy group or A group (P<0.05). Conclusion Intra-arterial thermochemoembolization may activate and improve cell-mediated immune function with a certain degree.

Objective To evaluate the value of two-dimension and color Doppler flow imaging(CDFI) in diagnosis of deep venous thrombosis(DVT) for different segments of lower extremity compared with digital subtract angiography(DSA). Methods Fifty-eight patients suspected of DVT enrolled in the study. Every segment venous of affected extremities was observed by CDFI. All patients underwent DSA examination. CDFI findings were compared with the results of DSA. Statistical analysis was according to quadruple tabular form. The indexes include sensitivity, specificity, accuracy, false positive rate and so on. Every segment of lower extremity deep venous was evaluated and analysis. Results Total of 150 vessels of fifty-eight patients were detected with CDFI and DSA. Evaluation findings of statistical analysis with quadruple tabular form as follows= sensitivity of common lilac vein was 82.6%, specificity was 66.7%; sensitivity of external lilac vein was 83.3%, specificity was 80.0%; sensitivity of hypogastria vein was 80.0 %, specificity was 66.7%;sensitivity of popliteal vein was 92.6 %, specificity was 92.2 % ;sensitivity of femoral vein was 96.2%, specificity was 93.8% ; sensitivity of anterior tibial vein was 71.4%, specificity was 75.0%; sensitivity of posterior tibial veins was 72.7%, specificity was 75.0%. Conclusions As the first-choice examination for DVT, ultrasonography has different diagnostic value for different segment of venous. It is more sensitive and accurate for femoral, popliteal and external iliac veins thrombus, but less sensitive for anterior,posterior tibial veins and bypass circuit vessels.