Objective To provide a technique of preparation recombinant hALR by gene engineering in larger scale.Methods Construct hALR(human augmenter of liver regeneration) protein expression vector pGEX-3X-hALR,induce the expreesion of hALR,and pruified the protein by affinity chromatography.Results Succeed in constructing the vector pGEX-3X-hALR and obtained the recombinant hALR protein.Conclusion The mothed could be used for preparation recombinant hALR by gene engineering in larger scale.

Objective To observe the regulation effect of transforming growth factor alpha(TGFα) on expression of glutamate transporter(GLAST)and ingestion activity of retinal M(u)ller cells in mice.Methods To take the retinal tissue of Kunming mouse at postnatal 7～10 day.and then cultured M(u)ller cells according to literature.The 3～4 generation cultured cells of the same primary cell were divided into two groups at random:①TGFα group:maintained in different concentrations of TGFα as 50,75,125 and 150 ng/ml,3 holes in each concentration;②Control group:cultured by Eagle culture medium which improved from Dulbeccon and contained 20%fetal calf serum.The influence of different concentrations TGFα on GLAST activity in M(u)ller cells were observed by L-3H-glutamate uptake detection;the expression of GLAST mRNA in M(u)ller cells was determined by RT-PCR;the expression of GLAST protein was detected with immunocytochemical staining.Results With the increase of TGFα concentration.both L-3H-glutamate uptake and GLAST mRNA expression were increased.The L-3H-glutamate accumulation had got to the maximum uptake at concentration of 125 ng/ml,which was 266% of that in control group,meanwhile,the expressions of GLAST mRNA also got to the maximum as 4 times of control group.Immunocytochemical staining indicated that the effect of 125ng/ml TGFα on expression of GLAST protein was higher than that in the control group,the differences between two groups were statistically significant(P＜0.05).Conclusion TGF-α can increase GLAST activity through up-regulating the expression of GLAST mRNA and protein.

Objective: To study the inhibition effect on Sarcoma 180 (S 180 ) and immune enhancing effect of rapeseed peptides (RSP) from enzymolysis of rapeseed protein. Methods: S 180 -bearing mice were used as animal model. The changes of tumor weight, lymphocyte proliferation, antibody in splenic cells and hemolysin were investigated. Results: Compared with cyclophosphamide (Cy), RSP inhibited S 180 growth markedly and enhanced immune function .Conclusion: RSP has potential in enhancing immune function and inhibiting tumor growth, but it deserves for further study.

Objective To study the distribution and tumor targeting property of anti-human prostate specific membrane antigen(PSMA) monoclonal antibody Ed-5 labeled with ~ 131 I in nude mice bearing human prostate cancer. Methods The nude mice xenografted models of human prostate cancer were established. Anti-human PSMA monoclonal antibody Ed-5 was labeled with ~ 131 I. ~ 131 I-Ed-5 was injected through the tail vein of nude mice bearing human prostate cancer. r-ray scintigraphy was performed at 24,48,96 and 144 hours after injection. And then the mouse were killed, the radioactivity ratio of tumor and non-tumor (T/NT), percentage of injected dosage of every gram tissue (%ID/g) were measured, and the bio-distribution of the labeled antibody was determined. Results The tumor imaging was clearly visualized at 96 hours after ~ 131 I-Ed-5 injection, and the tumor brim was clearer along with time prolonging. Radioactivity assembled in the tumors 24 to 144 hours after ~ 131 I-Ed-5 injection. The tumor's %ID/g at 96 hours after injection was 32.30, the radioactivity ratio of tumor and various tissues reached a maximum, and that of tumor/muscle was 7.08. Conclusion Ed-5 exhibited excellent immunoreactivity and tumor targeting property, and had application potential in targeting diagnosis and therapy of prostate cancer.

Objective By studying the expressions of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in rats′ cardiac muscle after acute myocardial infarction, we try to find out the relation between the expression of these two factors and the formation of new blood vessels under ischemia. Methods Wistar rats were divided into control group and infarction group (3, 7, 14, 28, 42, 56 d). The expression of VEGF and bFGF in cardiac muscle and endothelial cells is detected by means of immunohisto-chemial staining while the density of newly formed ressels is detected by marking the endothelial cells with antigens associated with factor Ⅷ. Results After ligating the left anterior descending coronary artery of the rats, the expression of VEGF and bFGF increased along with the prolongation of myocardial ischemia and reached the peak on the 7th day. The expression of the 2 factors began to decrease on the 28th day and the most significant decrease happened on the 42th and 56th day. The density of the newly formed capillaries is directly propontimal to the levels of the 2 factors. Less expression of VEGF and bFGF was observed in the control group. Conclusion The up-regulation of VEGF and bFGF expression might play important roles in neovascularization. Dicrectly intramyocardial injection of bFGF and VEGF gene at the time when the expression of bFGF and VEGF began to decrease maybe optimal.

BACKGROUND: The studies about cellular cardiomyoplasty (CCM) of bone marrow stromal cell (BMSC) are centered on modeling autologous cell transplantation while the study of myocardium transplantation of allogeneic BMSC is seldom reported domestically.OBJECTIVE: To study the hypothesis that the allogeneic BMSC, after transplanted into the myocardial infarction (MI) regions, can survive, further proliferate, differentiate, and its effects on host hearts.DESIGN: A randomized and controlled trial with experimental animals as subjects.SETTING: Laboratory of Internal Cardiology, Affiliated Union Hospital, Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: Ten Wistar rats of one-month old, body mass of 100-120 g and unconfined sex were used to culture BMSC while eighty female Wister rats of three-month old, body mass of 200-250 g were used for animal models.METHODS: The experiment was completed in the Laboratory of of Internal Cardiology, Affiliated Union Hospital, Tongji Medical College,Huazhong University of Science and Technology from June to December 2004. ①Eighty rats were used to establish the acute myocardial infarction (AMI) models by ligating the left anterior descending branch of coronary artery, and 45 of them were successful. ② After 4 weeks, the models were divided into 2 groups at random. In the experiment group (n=25), the passaged and uninduced BMSC cultured in vitro were injected into the MI region of the recipients while only medium was injected in the control group (n=20). ③At 4 weeks after implantation,the hemodynamic indexes of recipients' hearts were examined. Then the samples were obtained to detect the survival, differentiation and angiogenesis status of the removal cells.MAIN OUTCOME MEASURES: ①Comparison of hemodynamic indexes in rats of two groups②Results of the implantation cells③Changes of angiogenesis in rats of two groups.RESULTS: Totally 13 rats in the experiment group and 11 in the control group were involved in the result analysis. ①The hemodynamic indexes of rats were significantly improved in the experiment groups compared with the control group [Left ventricle systolic blood pressure (LVSBP): (88.61±5.99),(76.93±4.75) mm Hg, left ventricle end-diastolic pressure(LVEDP): (7.72±1.36),(12.77±2.76) mm Hg, P ＜ 0.05;The maximum changing velocity of LVSBP:(2 365.26±266.31),(2 025.04±230.25) mm Hg/s; The maximum changing velocity of LVDBP:(2 313.26±159.30),(2 140.12±191.03) mm Hg/s]. ②After implanted in the MI region, the allogeneic BMSC passed the acute inflammation period and did not induce the remarkable reject reaction of transplantation. The BMSC in the infarcted region were mainly differentiated into fibroblast. Some cells around the infarcted region were differentiated into endothelial cells, and improved the angiogenesis. ③The number of angiogenesis in and around the transplantation regions was significantly higher in the experiment group than in the control group (P ＜ 0.01).CONCLUSION: The allogeneic BMSC can not form cardiomyocyte in the infarcted region after cell transplantation, and the engendered endothelial cells of blood vessels may promote the angiogenesis after AMI and ameliorate the cardiac function.

To isolate and culture adipose stromal cells (ASCs), and study the effect of cytokines secreted by ASCs on endothelial cells, human adipose tissue was digested with collagenase type I solution and ASCs were derived by culture. The cells surface phenotype was examined by flow cytometry. ELISA was used to detect the secretion of VEGF, HGF, SDF-1 alpha and RT-PCR was employed to detect the expression of their mRNA. Then the ASC medium was utilized to culture human umbilical vein endothelial cells ECV304. Cells were counted by hemacytometer to determine the proliferation and Annexin V/ PI was employed for the examination of the apoptosis rate of ECV304. ASCs were derived by culture and expressed CD34, CD105 while they did not express CD31 or CD45. ASCs secreted cytokines such as VEGF, HGF and SDF-1 alpha so the ASC medium could stimulate proliferation and counteract apoptosis of endothelial cells (P < 0.05). Bcl-2 mRNA was also found to be up-regulated in the endothelial cells. It is concluded that ASCs can secrete cytokines and has significant effect on the proliferation of endothelial cells and apoptosis.

It has been suggested that progression of bladder transitional cell cancer (BTCC) may be regulated at the molecular level by a typical pattern of expression of genes involved in apoptosis. Recently Livin, belonging to the inhibitors of apoptosis (IAP) family, has been found to be expressed in most solid tumors, where its expression is suggested to have clinical significance. In order to explore the significance of Livin expression in the development of BTCC, immunohistochemistry and RT-QPCR were used to detect the expression of Livin mRNA in tumor tissues and adjacent normal tissues of 30 cases of BTCC. The results showed that the positive rate of Livin expression in adjacent normal tissues and tumor tissues was 0 and 60% (18/30) respectively. The-DeltaDeltaCT value of Livin in BTCC tissues was 8.0454 (7.4264-8.6644) times of that in adjacent normal tissues. The expression of Livin mRNA had no correlation with tumor pathological grades and clinical stages. It was suggested that there was weak expression of Livin mRNA in adjacent normal tissues, but strong in tumor tissues.

Objective: To study the effect of rapeseed peptides (RSP) from enzymolysis of rapeseed protein on growth of Sarcoma 180 (S180) cell incubated in vitro and its cell membrane. Methods: S180 cell incubated in vitro was used as model. The change of proliferation of S180 cell, fatty acid and sialic acid in membrane and membrane fluidity of S180 cell were investigated. Results: RSP had inhibitiory ability to S180 cell proliferation and could influence its membrane. Conclusion: RSP had potential in inhibiting tumor growth, so was needed to study further.

Objective:To compare the anti tumor activity and immune enhancing effect of crude(PGF) and purified (PGF 1) polysaccharide from Grifola frondosa. Methods:S 180 bearing mice were used as animal model. The effects on tumor weight and immune function were investigated. Results:PGF and PGF 1 showed inhibitory activity on S 180 growth. They could also increase weight of immune organs and improved the phagocytic function, DTH response, lymphocyte proliferation, antibody formation in splenic cells and content of serum HC IgM markedly. PGF was better than PGF 1. Conclusion:Both PGF and PGF 1 can inhibit S 180 tumor growth and enhance immune function in S 180 bearing mice.