Expression of sphingosine kinase gene in the interactions between human gastric carcinoma cell and vascular endothelial cell

AIM：To study the interactions between human gastric carcinoma cell(HGCC)and human vascular endothelial cell(HVEC),and if the expression of sphingosine kinase (SPK)gene was involved in these interactions.METHODS:The specific inhibitor to SPK,dimethyl sphingosine(DMS),was added acting on HGCCand HVEC,then the cell proliferation was measured by MTT.The conditioned mediums(CMs)of HGCCand HVEC were prepared.The CMof one kind of cell was added to the other kind of cell,and the cell pro,liferation was measured by MTT,After the action of CM,the cellular expression of SPK gene in mRNA level was detected with in situ hybridization(ISH).RESULT:DMS could almost completely inhibit the proliferation of HGCCand HVEC.The growth inhibitory rates could amount to 97.21%.83.42%.respectively(P<0.01).The CMof HGCCcould stimulate the growth of HVEC(2.70±0.01,P<0.01)while the CMof HVEC could inhibit the growth of HGCC(52.97±0.01%,P<0.01).There was no significant change in the mRNA level of SPK gene in one kind of cell after the action of the CMof the other kind of cell.CONCLUSION:SPK plays a key role in regulating the proliferation of HGCCand HVEC.There exist complicated interactions between HGCCand HVEC.HGCC can significantly stimulate the growth of HVEC while HVEC can significantly inhibit the growth of HGCC.The expression of SPK gene is not involved in the interactions.     

The role of KDR in the interactions between human gastric carcinoma cell and vascular endothelial cell

AIM：To study the interactions between human gastric carcinoma cell(HGCC)and human vascular endothelial cell(HVEC),and the role of KDRin these interactions.METHODS:Antisenes oligodexynucleotide(ASODN)specific to KDR gene was devised and added to the culture medium of HGCC and HIVEG.After the action of ASODN,the proliferation of two cells was mesured by MTTmethod.The role of KDR in regulating the proliferation of two kinds of cells was known through oroliferation of two kinds of cells was known through observing the effect of ASODN on them.The conditioned mediums(CMs)of HGCCand HVEC were prepared.The CMof one kind of cell was added acting on the other kind of cell,then the cell proliferation was measured byMTT,After the action of ASODNor CM,the cellular expression of KDR gene was detected with in situ hybridization(ISH)for mRNA level and with immunohistochemical staining for protein level.ABC-ELISA was used to detect hVEGFin the CMs of two cells.RESULTS:KDR ASODN could specifically inhibit the proliferation of HGCCand HVEC significantly.The growth inhibitory rate amounted to 55.35%and 54.83%,respectively(P<0.01).HGCC and HVECcould secreta certain level of hVEGF(92.06±1.69ng/L.77.70±8.04ng/L).The CMof HGCCcould significantly stimulate the growth(2.70±0.01times)and KDRgene expression of HVEC(P<0.01)while the CM of HVEC could significantly inhibit the growth(52.97±0.01%)and KDRgene expression of HGCC(P<0.01).CONCLUSION:KDR plays a key role in regulating the proliferation of HGCC and HVEC.There exist complicated interactions between HGCCand HVEC.HGCCcan significantly stimulate the growth of HVE while HVEC can significantly inhibit the growth of HGCC.KDRis involved in the interactions between them.     

Liver fibrosis and tissue architectural change measurement using fractal-rectified metrics and Hurst＇s exponent

AIM: To provide the accurate alternative metrical means of monitoring the effects of new antiviral drugs on the reversal of newly formed collagen. METHODS: Digitized histological biopsy sections taken from 209 patients with chronic C virus hepatitis with different grade of fibrosis or cirrhosis, were measured by means of a new, rapid, user-friendly, fully computer-aided method based on the international system meter rectified using fractal principles. RESULTS: The following were described: geometric perimeter, area and wrinkledness of fibrosis; the collation of the Knodell, Sheuer, Ishak and METAVIR scores with fractal-rectified metric measurements; the meaning of the physical composition of fibrosis in relation to the magnitude of collagen islets; the intra- and inter-biopsy sample variability of these parameters; the "staging" of biopsy sections indicating the pathway covered by fibrosis formation towards its maximum known value; the quantitative liver tissue architectural changes with the Hurst exponent. CONCLUSION: Our model provides the first metrical evaluations of the geometric properties of fibrosis and the quantitative architectural changes of the liver tissue. The representativeness of histological sections of the whole liver is also discussed in the light of the results obtained with the Hurst coefficient.     

血管超声对椎动脉闭塞的血流动力学评价

Objective To investigate the combination of color Doppler flow imaging (CDFI) and transcranial Doppler (TCD) in the assessment of the hemodynamic changes of vertebral artery occlusion disease and their clinical value. Methods A total of 101 patients with vertebral artery occlusion detected by the combination of CDFI and TCD and confirmed by DSA were enrolled from January 2005 to January 2009. Taking the result of digital subtraction angiography (DSA) as a golden standard, The differences between the different types of the side of vertebral artery occlusion and contralateral vertebral artery on the extra- and intracranial segments were compared and analyzed in peak systolic velocity (PSV), end diastolic velocity (EDV), resistive indices (RI), pulsatility index (PI), spectrum morphology, and hemodynamics. Results No blood flow signals were detected by CDFI and TCD in patients of complete occlusion of the vertebral artery; the weak blood flow signals were detected by CDFI in patients of the occlusion in the intracranial segment of the vertebral artery. 3he blood flow signals after the establishment of collateral circulation in patients of the segmental occlusion were detected in the extraeranial segment or intraeranial segment of the vertebral artery. The PSV on the occluded sides of the extraeranial segments were decreased more significantly than that on the unoccluded sides (27.39 ± 12.44 cm/s vs. 62.61 ± 13.22 cm/s, P = 0.000); RI was significantly higher than the unoccluded sides (0. 99 ± 0. 21 vs. 0. 62 ± 0.07, P = 0. 000). When a vertebral artery had the segmental occlusion and the collateral circulation was established, the PSV, EDV, and PI of the intracranial segment of the vertebral artery on the occluded sides were decreased more significantly than those on the unoccluded sides, PSV were 37. 81 ± 12.28 cm/s and 73.17±30. 99 cm/s, respectively (P =0. 000), EDV were 17.58 ± 7.10 cm/s and 29.31 ± 12.94 cm/s, respectively (P = 0. 000), PI were 0.84 ± 0.22 and 1.01 ±0. 18, respectively (P=0. 000). The compard DSA showed that the sites of vertebral artery occlusion were different, There was significant difference in the Doppler flow velocity spectrum between the CDFI and TCD. Conclusions PSV, EDV, RI, and PI are the hemodynamic parameters of accurately assessing vertebral artery occlusion, and the combination of CDFI and TCD has significant value for the hemodynamie changes of different types of vertebral artery occlusion and the clinical comprehensive assessment.     

Influence of hepatic arterial blockage on blood perfusion and VEGF,MMP—1 expression implanted liver cancer in rats

AIM：To investigate the influence of hepatic arterial blodckage on blood perfusion of transplanted cancer in rat liver and the expression of vascular endothelial growth factor (VEGF)and matrix metalloproteinase-1(MMP-1),and to explore the mechanisms involved in transarterial embolization(TAE)-induced metastasis of liver cancer preliminarily.METHODS:Walker 256 carcinosarcoma was transplanted in to rat liver to establish the liver cancer model.Hepatic arterial ligation(HAL) was used to block the hepatic arterial blood supply and simulate TAE.Blood perfusion of tumor in control laparotomy control,and HAL group was analyzed by Hoechst 33342 labeling assay,the serum VEGF level was assayed by ELISA,the expression of VEGF and MMP-1 mRNA was detected by in situ hybridization.RESULTS:Two days after HAL.the number of Hoechst 33 342 labeled cells which represent the blood perfusion of tumor directly and hypoxia of tumor indirectliy in HAL group decreased significantly compared with that in control group (329±29VS 384±19,P<0.01),The serum VEGF level in the HAL group increased significantly as against that of the control group (93ng.L^-1±44ng.L^-1VS55ng.L^-1±19ng.L^-1,P<0.05),The expression of VEGFand MMP-1 mRNA in the tumor tissue of the HAL group increased significantly compared with that of the control and the laparotomy control groups(P<0.05),The blood perfusion data of the tumor represented by the number of Hoechst 33342 labeled cells,showed a good linear inverse correlation with the serum VEGF level(r=-0.606,P<0.05),and the expression of VEGF mRNA in the tumor tissue(r=-0.338,P<0.01).CONCLUSION:Blockage of hepatic arterial blood supply results in decreased blood perfusion and increased expression of metastasis-associated genes VEGF and MMP-1 of transplanted liver cancer in rats .Decreased blood Perfusion and hypoxia may be the major cause of upregulated expression of VEGF.±±     

Effects of epinephrine and glucose on the in vitro response of adipose tissue to sulfonylureas

Summary Pieces of lumbar fat pads from fed female rats were incubated in the presence of 1-¹⁴C-glycerol. Epinephrine (2.8 μM), glucose (5 mM), tolbutamide, glipentide or glibenclamide were added to the incubation media. The sulfonylurea drugs reduced the lipolytic effect of epinephrine in the absence of glucose; the effect of glibenclamide was greatest. In the absence of epinephrine, both glipentide and glibenclamide reduced the uptake of 1-¹⁴C-glycerol and its conversion to glyceride-glycerol while tolbutamide had no effect on these parameters. In the presence of glucose, the lipolytic effect of epinephrine was retained but tolbutamide and glipentide no longer affected glycerol metabolism. Glibenclamide still inhibited the production of glycerol and the utilization of 1-¹⁴C-glycerol by the tissues in the presence of glucose. The possibility that sulfonylureas exert their effect on adipose tissue glycerol metabolism by acting as uncoupling agents is discussed.     

Islet isolation assessment in man and large animals

Summary Recent progress in islet isolation from the pancreas of large mammals including man, accentuated the need for the development of precise and reproducible techniques to assess islet yield. In this report both quantitative and qualitative criteria for islet isolation assessment were discussed, the main topics being the determination of number, volume, purity, morphologic integrity andin vitro andin vivo function tests of the final islet preparations. It has been recommended that dithizone should be used as a specific stain for immediate detection of islet tissue making it possible to estimate both the total number of islets (dividing them into classes of 50 μ diameter range increments) and the purity of the final preparation. Appropriate morphological assessment should include confirmation of islet identification, assessment of the morphological integrity and of the purity of the islet preparation. The use of fluorometric inclusion and exclusion dyes together have been suggested as a viability assay to simultaneously quantitate the proportion of cells that are intact or damaged. Perifusion of islets with glucose provides a dynamic profile of glucose-mediated insulin release and of the ability of the cells to down regulate insulin secretion after the glycemic challenge is interrupted. Although perifusion data provides a useful guide to islet viability the quantity and kinetics of insulin release do not necessarily predict islet performance after implantation. Therefore, the ultimate test of islet viability is their function after transplantation into a diabetic recipient. For this reason,in vivo models of transplantation of an aliquot of the final islet preparation into diabetic nude (athymic) rodents have been suggested. We hope that these general guidelines will be of assistance to standardize the assessment of islet isolations, making it possible to better interpret and compare procedures from different centers.     

Failure of ergotamine to act as an amplifier of sulfonylurea-stimulated insulin secretion

Summary Normal dogs were injected i.v. with 0.25 mg/kg sodium salt of HB 419 (gliben-clamide) and plasma insulin concentrations were measured over a period of 2 hrs. When the animals were given a single i.v. injection of 0.2 mg/kg dihydroergotamine tartrate (DHE) 30 min prior to the administration of HB 419, the insulinogenic effect of the sulfonylurea was considerably amplified (192 μU/mlvs 34 μU/mI at 45 min). No augmentation of the insulinogenic effect of HB 419 was observed when the same experiments were conducted with 0.05, 0.025 or 0.01 mg/kg ergotamine tartrate. At the dose level of 0.1 mg/kg the insulinogenic effect of HB 419 was suppressed. Since the structural difference between these two ergor alkaloids consists of the presence or absence of the double bond at C₉ and C₁₀ of the lysergic acid moiety, it appears that saturation of this double bond is an essential structural requirement for DHE to function as an amplifier of sulfonylurea-stimulated insulin release. Supported by the Karel Ančerl Fund of the University of Toronto.     

Stereologic study of the sinoatrial node of rats -- age related changes

Changes in the sinoatrial node represent the major mechanism of sudden death in humans, and because of the sparse knowledge about the effects of aging on this structure, light microscopic and quantitative studies of the sinoatrial node were undertaken. Twenty-one hearts were studied, seven rat hearts from each of the following age groups: three months of age, twelve months of age and eighteen months of age. In the stereo logic study, the following parameters were studied: Vv_[nc] and Vv_{[interstitium]} % (the volume densities of the nodal cell and interstitium, determined by the point-couting method), and Nv_[nc](1/mm³) (the numerical density of the nodal cell, determined by the disector method). The mean volume of the nodal cell (V_[nc]) (μm³) was also determined. The comparisons showed that in the oldest animals, the volume density of the nodal cells decreased, while the volume density of the interstitium increased. Although numerical density of the nodal cell per volume of sinus node decreased, the nodal cells displayed increased mean volume with age. In conclusion, the aging process implies changes in the cell and fiber content of the sinoatrial node. This revised version was published online in July 2006 with corrections to the Cover Date.     

The study between the dynamics and the X-ray anatomy and regularizing effect of gallbladder on bile duct sphincter of the dog

AIM: To study the relationship between the radiological anatomy and the dynamics on bile duct sphincter in bile draining and regulatory effect of gallbladder.METHODS: Sixteen healthy dogs weighing 18 kg to 25 kg were divided randomly into control group and experimental group (cholecystectomy group). Cineradiography, manometry with perfusion, to effect of endogenous cholecystokinin and change of ultrastructure were employed.RESULTS: According to finding of the choledochography and manometry, in control group the intraluminal basal pressure of cephalic cyclic smooth muscle of choledochal sphincterc CS was 9.0±2.0mmHg and that of middle oblique smooth muscle of choledochal sphincter (mOS) was 16.8±0.5mmHg,the intraluminal basal pressure of cCS segment was obviously lower than that of mOS (P<0.01) in the interval period of bile draining, but significant difference of intraluminal basal pressure of the mOS segment was not found between the interval period of bile draining (16.8±0.5mmHg) and the bile flowing period (15.9±0.9mmHg) (P>0.05). The motility of cCS was mainly characterized by rhythmically concentric contraction, just as motility of cCS bile juice was pumped into the mOS segment in control group. And motility of mOS segment showed mainly diastolic and systolic activity of autonomically longitudinal peristalsis. There was spasmodic state in cCS and mOS segment and reaction to endogenous cholecystokinin was debased after cholecystectomy. The change of ultrastructure of cCS portion showed mainly that the myofilaments of cell line in derangement and mitochondria is swelling.CONCLUSION: During fasting, the cCS portion has a function as similar cardiac “pump” and it is main primary power source in bile draining, and mOS segment serves mainly as secondary power in bile draining. The existence of the intact gallbladder is one of the important factors in guaranteeing the functional coordination between the cCS and mOS of bile duct sphincter. There is dysfunction in the cCS and mOS with cholecystectomy.     

In vitro effect of exogenous insulin on insulin secretion. Studies with glucose,leucine, arginine,aminophylline and tolbutamide

Summary Using rat pancreatic islets and the perfused rat pancreas, the effect of exogenous insulin on insulin secretion mediated by glucose, leucine, arginine, aminophylline and tolbutamide was studied. (1) In both systems the insulin releasing capacity of glucose was inhibited by exogenous insulin. In the perfused pancreas the inhibition concerned the first and the second phase of insulin release; (2) the EC50 (half-maximal inhibitory effect of insulin on glucose-induced insulin secretion) in islets was 1.2 nM (=160 μU/ml) and 2.8 nM (390 μU/ml) in perfused pancreas; (3) exogenous insulin also inhibited insulin release in response to leucine and arginine in the isolated islet system and in the perfused pancreas; (4) using aminophylline and tolbutamide in combination with glucose, the extent of the inhibitory effect of insulin was in the range of the inhibitory effect when glucose was used alone as stimulator in islets. Data suggest that the insulinogenic action of physiological stimulators including glucose, leucine and arginine is inhibited by exogenous insulin whereas this seems not to be the case when insulin release was stimulated by aminophylline and tolbutamide. Comparing the EC50s, isolated islets seem to be more sensitive to inhibition than the perfused pancreas when glucose was used as stimulator. As far as glucose is concerned the inhibitory effect seems to depend on the extent of its concentration and/or the extent to which the mechanism of insulin release is sensitive to stimulation. The EC50 of the inhibitory effect of exogenous insulin was in the range of dissociation constant of binding of insulin to insulin receptors of islets. Preliminary data on this study have been presented at the 22th Spring Meeting of theDeutsche Pharmakologische Gesellschaft, Mainz, F.R.G., March 10–13, 1981. This study was supported by grants of theDeutsche Forschungsgemeinschaft, Bonn-Bad Godesberg, F.R.G.     

Effect of cholecystokinin on experimental neuronal aging

AIM: To observe the effect of cholecystokinin (CCK) on lipofusin value, neuronal dendrite and spine ultrastructure, and total cellular protein during the process of experimental neuronal aging. METHODS: Experimental neuronal aging study model was established by NBA2 cellular serum-free culture method. By using single intracellular lipofusin value from microspectrophotometry, morphology of neuronal dendrites and spines from the scanner electron microscopy, and total cellular protein as the indexes of experimental neuronal aging, we observed the effect of CCKs on the process of experimental neuronal aging. RESULTS: Under the condition of serum-free culture, intracellular fluorescence value (%) increased with the extension of culture time (1 d 8.51±3.43; 5 d 10.12±3.03; 10 d 20.54±10.3; 15 d 36.88±10.49; bP<0.01). When CCK was added to serum-free culture medium, intracellular lipofusin value (%) decreased remarkably after consecutive CCK reaction for 10 and 15 d (control 36.88±10.49; 5 d 32.03±10.01; 10 d 14.37±5.55; 15 d 17.31±4.80; bP<0.01). As the time of serum-free culturing was prolonged, the number of neuronal dendrite and spine cells decreased. The later increased in number when CCK8 was added. CCK8 could improve the total cellular protein in the process of experimental neuronal aging. CONCLUSION: CCKs may prolong the process of experimental neuronal aging by maintaining the structure and the number of neuronal dendrite and spine cells and changing the total cellular protein.     

Preparation of monoclonal antibody against apoptosis—associated antigens of hepatoma cells by subtractive immunization

AIM：To elucidate the expression of the apoptosis-associated molecules in human primary hepatocellular carcinoma(HCC)cells,and prepare the monoclonal antibodies(mAb)against the apoptosis-associated antigens of HCC cells.METHODS:Human HCC cell line HCC-9204 cells were induced apoptosis with 60mL&#183;L^-1 ethanol for 6h and their morphological changes were obserevd by transmission electron microscope,The cell DNA fragmentations were detcted by Terminal Deoxynucleotidyl transferase-mediated dUTPnick end labeling(TUNEL)assay,and the cell DNA contents by flow cytometry.Ten mice were immunized with ethanol-induced apoptoticHCC-9204 cells with the method of subtractive immunization.while the other 10mice used as the control were immunized by the routin procedures.The tail blood of all the mice were prepared after the last immunization.and the produced antibodies were determined by the immunocytochemical ABC staining .The splenic cells of the mice whose tail blood sera-HCC-9204 cells serum reactions were most different between the apoptotic and the non-apoptotic were prepared an fused with the mouse myeloma cell lineSP2/0cells.The positive antibodies were selected by ELISA assay.The fusion rates of hybridona cells and the producing rates of antibodies were calculated.The fused cells that secreted candidate objective antibody were cloned continually with the of limited dilution method.and then selected and analyzed further by the immunocytochemical ABCstaining.The chromosomes of the cloned hybridoma cells that secreted objective mAb and the mAb immunoglobulin(Ig)subtpe of the prepared mAb were also determined.The molecular mass of the mAb associated antigen was analyzed by Western blot assay.RESULTS:HCC-9204 cells treated with60mL&#183;L^-1 ethanod for 6h,manifested obvious apoptotic morphological changes,the majority of the cells wereTUNEL-positive,and the sub-G1 apoptotic peak was evident.There were 2mice in the experimental group whose tail blood serum reacted strongly with the apoptotic HCC-9204 cells,but weakly with their non-apoptotic counterparts,In the fusion rates of hybridoma cells as well as the producing rates of the antibody deseribed above,there did not show significant difference between the experimental and the control group,but weakly with non-apoptoticHCC-9204.However,the total producing rate of antibodies in the experimental group wa significantly lower compared with the control(P&lt;0.01).and so was the producing rate of the Antibodies which racted strongly with both paoptotic and non-apoptotic HCC-9204cells(P&lt;0.01).After cloned continually for several times the cell that produce mAb which reacted strongly with the nuclei of ethanol-induced apoptotic HCC-9204cells,but very weakly with that of non-apoptotic cells was selected out.Chromosme analysis revealed that the selected cell was with the universal characteristics of the monoclonal hybridoma cells which secreted mAb,and the Ig subtype of the prepared mAb was IgG1,The molecular mass of this mAb associated antigen of was about75ku.CONCLUSION :Subtractiv immunization is a useful method to prpar the mAb against the apoptosis-associated antigens of cells,The expression of some molecules increases to some extent in HCC-9204cells in the process of apoptosis induced by low-concentration ethanol.The mAb that may be against ethanol-induced apoptosis-associated antigens of HCC cells was successfully prepared and primarily identified.     

Bax protein may influence the invasion of colorectal cancer

AIM: To evaluate the expression of Bcl-xL, Bak, and Bax proteins in correlation with particular clinico-histopathological parameters, including tumor invasion front, in patients with colorectal cancer. METHODS: The expression of these proteins was evaluated with the use of the immunohistochemical method in 50 primary tumors. RESULTS: According to observations, a low expression of Bax and Bak proteins is related to the localization of the tumor in the rectum(P < 0.05 and P < 0.05 respectively), which may explain an increased incidence of colorectal cancer in this area. A positive expression of Bax protein also correlates with the presence of cancer cell infiltration to lymph and blood vessels(P < 0.05), which may suggest the participation of this protein in the early stages of colorectal cancer progression. Moreover, a positive expression of Bcl-xL protein correlated with a positive expression of Bak protein. This may suggest a greater participation of Bcl-xL protein in the inhibition of the proapoptotic Bak protein, but not the Bax protein. CONCLUSION: Bax protein is probably very significant in the cancerogenesis mechanism in the large intestine.     

Experimental assessment of a modified retrograde traction tracheal intubation method for increasing the success rate of myocardial infarction model in rats

Background Animal models of myocardial infarction （MI） have been widely used to study the pathologi- cal and physiological changes that occur in MI, and to objectively evaluate the efficacy of new treatments. They are an important tool in this procedure. However, the mortality rate of MI animal models has so far been higher than in real-life situations. The aim of this study was to explore the use of a modified retrograde traction tracheal intubation （MRTI） method for increasing the success rate of MI models in rats. Methods Sixty male Sprague-Dawley rats were used in the experiment. Using the MRTI method of artificial airway generation, we established the MI model by ligation of the left anterior descending branch of the coronary artery. We analyzed the effects of MRTI, the use of lidocaine, operative details, nursing considerations during the operation, and post-operative factors on the success rate of the MI model in rats. Results The success rate of generating an MI model in rats can be significantly increased using the following methods： 1） Setting up the artificial airway through the use of MRTI by using a single-lumen central venous catheter; 2） Selecting a ligation site 2 mm be- low the midpoint of the connection between the left atrial appendage and the pulmonary cone; 3） Adding a drop of lidocaine to the surface of the heart to slow down the heart rate, make the operation easier to perform, and prevent arrhythmias postoperatively; 4） Clearing up airway secretions timely both intra and postoperative- ly; 5） Making sure that rats are in a warm state both intra and postoperatively; 6） Preventing wound infection. Conclusions Use of the MRTI method can quickly establish an artificial airway in rats. Intraoperative use of lidocaine, selecting a precise vascular ligation site, and appropriate care both intra and postoperatively can in- crease the success rate of MI model generation.     

Effect of fermented soy milk on the intestinal bacterial ecosystem

AIM: To investigate the effect of fermented soy milk on human ecosystem in the intestinal tract by way of examining the population of different microorganisms isolated from fecal samples. METHODS: A crossover experimental design was applied. Twenty-eight healthy adults completed this experiment. Each subject consumed 250 ml, twice a day between meals, of either fermented soy milk or regular soy milk first for 2 wk, then switched to the other drink after 2 wk. Fecal samples were collected from all subjects every week starting from the second week to the end of the experiment. The microorganisms analyzed were Bifidobacterium spp., Lactobacillus spp., Clostridium perfringens, coliform organisms, and total anaerobic organisms. RESULTS: In the period of fermented soy milk consumption, the populations of Bifidobacterium spp. and Lactobacillus spp. increased (P<0.05) as well as the ratios of Bifidobacterium spp. and Lactobacillus spp. to Clostridium perfringens (P<0.05). The population of coliform organisms decreased (P<0.05) when subjects were in the period of fermented soy milk consumption. CONCLUSION: Intake of fermented soy milk significantly improved the ecosystem of the intestinal tract in the body by increasing the amount of probiotics.     

Effect of tetramethylpyrazine on exocrine pancreatic and bile secretion

AIM: To investigate the effect of tetramethylpyrazine ?A (ligustrazine, TMP) on the secretion of exocrine pancreas(and biliary). METHODS: In in vivo study, we investigated the effect of TMP on the secretion of pancreatic-bile juice (PBJ) in rats. Using human pancreatic duct cell line, CAPAN-1, combined with the short-circuit current (Isc) technique we further studied the effect of TMP on the pancreatic anion secretion. RESULTS: Administration of TMP (80mg/kg, ip) significantly increased the secretion of PBJ (P&lt;0.05), but the pH of PBJ and the secretion of pancreatic protein were not significantly affected. Basolateral addition of TMP produced a dosedependent increase in Isc(EC50=1.56mmol/L), which contained a fast transient Isc response followed by a slow decay. Apical application of CI- channel blockers, DPC (1mmol/L), decreased the response by about 67.1% (P&lt;0.001), whereasamiloride (100μmol/L), a epithelial sodium channel blockers, had no effect. Removal of extracellular HCO3^- abolished TMP-induced increase in Isc by about 74.4% (P&lt;0.001),but the removal of external Cl^- did not. Pretreatment with phosphodiesterase inhibitor, IBMX(0.5mmol/L), decreasedthe TMP-induced Isc by 91% (P&lt;0.001). CONCLUSION: TMP could stimulate the secretion of PBJ, especially pancreatic ductal HCO3- secretion via cAMP or cGMP-dependent pathway. It need further study to investigate the roles of cAMP or cGMP in the effect of TMP on the secretion of exocrine pancreas.     

Potent inhibition of angiogenesis and liver tumor growth by administration of an aerosol containing a transferrin-liposome-endostatin complex

AIM: To obtain an efficient delivery system for transporting endostatin gene to mouse liver tumor xenografts by administration of aerosol.METHODS: Recombinant plasmid pcDNA3.0/endostatin containing human endostatin gene together with signal peptide from alkaline phosphatase were transferred into human umbilical vein endothelial cell (HUVEC) by transferrin(TF)-Iiposome-endostatin complex. Western blot was used to detect the expression of human endostatin in transfected HUVEC cells and its medium. After the tumor-bearing mice were administrated with TF-liposome-endostatin complex,the lung tissue was analyzed by immunohistochemical method for expression of endostatin and the tumors were treated with CD-31 antibody to detect the density of microvesseles in tumor tissues. The inhibition of tumor growth was estimated by the weight of tumors from groups treated with different doses of TF-liposome-endostatin complex. DNA fragmentation assay was used to detect the apoptosis of the cells from primary liver tumor.RESULTS: Western blot analysis and immunohistochemical method confirmed the expression of endostatin protein in vitro and in vivo. After the tumor sections were treated with CD-31 antibody, the positive reaction cells appeared brown while the negative cells were colorless. The positively stained area of the TF-liposome-endostatin treated group was significantly smaller (P&lt;0.01, 645.8+55.2 μm^2) than that of the control group (1325.4&#177;198.5 μm^2). The data showed a significant inhibition of angiogenesis. After administration of TF-liposome-endostatin, comparing with the control group administrated with TF-liposome-pcDNA3.0, liver tumor growth in the mice treated with 50, 250 and 500 mg DNA/kg was inhibited by 36.6 %, 40.8%, and 72.8%, respectively(P&lt;0.01). And a typical DNA fragmentation of apoptosis was found in the cells from tumor tissues of the mice treated with TF-liposome-endostatin but none in the control group.CONCLUSION: Endostatin gene could be efficiently transported into the mice with TF-liposome-DNA delivery system by administration of aerosol. TF-liposome-mediated endostatin gene therapy strongly inhibited angiogenesis and the growth of mouse xenograft liver tumors. It also could promote the development of apoptosis of tumors without direct influence on tumor cells.     

Effect of lactulose on establishment of a rat non-alcoholic steatohepatitis model

AIM: To explore the relationship between changes of intestinal environment and pathogenesis of non-alcoholic steatohepatitis (NASH). METHODS: Forty-two Sprague-Dawley rats were randomly divided into model group (n = 24), treatment group (n = 12), and control group (n = 6). The rats of model and treatment groups were given high-fat diet, and those of the control group were given normal diet. Furthermore, the rats of treatment group were given lactulose after 8 wk of high-fat diet. Twelve rats of the model group were killed at 8 wk of high-fat diet. At the 16 wk the rats of treatment group, control group, and the rest of the model group were killed. The serum levels of aminotransferase were measured and the histology of livers was observed by H&E staining. RESULTS: The livers of rats presented the pathological features of steatohepatitis with higher serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the model group after 16 wk. Compared to the model group, the serum levels of ALT and AST in treatment group decreased significantly and were close to the normal group, and the hepatic inflammation scores also decreased markedly than those in the model group after 16 wk (5.83±2.02 vs3.63±0.64, P<0.05), but were still higher than those in the model group after 8 wk (3.63±0.64 vs 1.98±0.90, P<0.05). However, the degree of hepatic steatosis had no changes in treatment group compared to the model group after 16 wk. CONCLUSION: Lactulose could ameliorate the hepatic inflammation of rats with steatohepatitis induced by fat-rich diet, but could not completely prevent the development of steatohepatitis. It is suggested that intestinal environmental changes such as intestinal bacteria overgrowth, are one of the important factors in the pathogenesis of NASH.     

Studies on the Relationship between Production of Oxygen Free Radicals and Cardiac Failure of Coronary Artery Disease

<正> In this paper, the relationship between Oxygen Free Radicals（OFR） and cardiac failure of Coronary Artery Disease（CAD）was studied. The authors analyse the release of OFR, as measured by Lumin-dependent chemiluminescence, from the polymorphonuclear leukocytes（PMN）; the activity of plasma superoxide dismutase （SOD） and the concentration of malondialdehyde（MDA）in patients of CAD with heart failure. The results showed that compared with the health controls and the group of CAD without heart failure, the release of OFR in respiratory burst of the PMN stimulated with PMA（phorbol myristate acetate）and the concentration of MDA in plasma were significantly increased（p<0.05） and the activity of SOD was markedly lower（p<0.05）in the group of CAD with heart failure.It suggests that the enhanced production of OFR by PMN, the increased concen tration MDA in plasma and the decreased SOD might be associated with the genesis and development of cardiac failure, The use of agents that reduce the amount of OFR would be of value in the prevention and treatment of cardiac failure.