Identification and application of sphingosine-1-phosphate receptor of human umbilical vein endothelial cells / 天津医药

Objective To identify the expression of sphingosine-1-phosphate receptor (S1PR) subtypes, C-myc and His tag proteins of human umbilical vein endothelial fusion cell line, EA.hy926 and human umbilical vein endothelial cells (HUVEC), CRL-1730 for studying the function of apolipoprotein M (ApoM)-S1P axis. Methods Two kinds of cells (EA. hy926 and CRL-1730) were cultured to reach the density of 60%-70% in vitro. Immunofluorescence technique was em?ployed to investigate the expressions of coagulation factorⅧ(FⅧ), ApoM, S1PR1-S1PR5, C-myc and His tag proteins. Re?sults (1) Two kinds of cells both expressed FⅧand ApoM. FⅧpresented scattered particle distribution in CRL-1730, while uniform distribution in EA.hy926. However, ApoM was strongly expressed and widely distributed in cytoplasm of two kinds of cells. (2) S1PR1-3 can be detected on their membrane other than S1PR4 and S1PR5. S1PR1 was highly expressed but S1PR2 and S1PR3 were in a low level expression. (3) Two kinds of cells both expressed C-myc and His tag proteins in cytoplasm. Conclusion Two kinds of cells have the properties of endothelial cells and can express FⅧ, ApoM, C-myc and His tag proteins. It is not suitable for choosing C-myc and/or His tag–conjugated recombinant ApoM to study the fuction of ApoM-S1P axis with these two kinds of cells.

Protective Effects of Extract of Periplaneta Americana on Liver Fibrosis in Rats / 天津医药

Objective To investigate the protective effects of extract of Periplaneta americana (APA) against carbon tetrachloride (CCl4)-induced hepatic fibrosis in rats. Methods Seventy SD rats were divided into five groups:normal con-trol group (C), model control group (M), extract of APA 1 group (APA1), extract of APA 2 group (APA2) and reduced glutathi-one group (R). The liver fibrosis model was induced by injecting 40%CCl4-olive solution subcutaneously for seven weeks in M, APA1, APA 2 and R groups. Drugs were given at the same time. Rats were killed at 7-week (C, M, APA1 and R groups), and were killed at 9-week (APA2 group). The serum values of transaminase (ALT, AST), albumin (ALB), hyaluronic acid (HA) and laminin (LN), and hepatic inflammation and fibrosis were detected respectively. The expressions of apoptotic relat-ed gene Bcl-2 and Bax protein in central veins and portal areas of hepatic lobules were detected by immunohistochemical method. Results Rats showed poor nutritional status, and significantly increased transaminase, HA and LN in M group, but the serum level of ALB was significantly lower than that in C group. There was extensive necrosis of liver cells, obviously fi-brosis or cirrhosis in model rats’liver tissues. The serum contents of ALT, AST, HA and LN were significantly decreased, the serum level of ALB were significantly increased in APA1,APA2 and R groups (P<0.05). There were complete hepatic lobule and no obvious fibrous tissue hyperplasia in liver biopsy of APA1 and R groups. The values of Bcl-2/Bax proteins in central vein of liver tissues were significantly higher in APA1,APA2 and R groups than those of M group (P<0.05), but the ratio of Bcl-2/Bax was lower in periportal area of liver tissues (P<0.05). There were significantly higher body weight and ALB levels in APA1 group than those in R group (P<0.05). The level of transaminase was slightly higher in APA1 group than that of R group. The degeneration of liver cells was found mostly in R group. Conclusion The extract of Periplaneta americana has a role in protecting liver cells, inhibiting the progression of hepatic fibrosis, and improving the nutritional sta-tus of trial rats.

Effects of inactivation of vitamin D receptor in intestinal tumor growth of APCmin/+ mice / 中国医师杂志

Objective To explore effects and mechanism of inactivation of vitamin D receptor (VDR) in intestinal tumor growth of APCmin/+ mice. Methods To generate APCmin/+VDR -/- mice through breeding, the number and size of small intestinal and colonic tumors were assessed and compared between APCmin/+ and APCmin/+ VDR -/- mice. The intestinal tumors were diagnosed with HE stain. The expressions of BCL-2,vimentin-1,Stat-1 and MSH-2 proteins of tumors were determined by immanohistocbemistry and compared be-tween APCmin/+ and APCmin/+VDR -/- mice. Results A comparison between APCmin/+ and APCmin/+ VDR -/- mice revealed that the number of the tumors, which were larger than 3mm, was significantly increased in APCmin/+ VDR -/- mice at 4 months of age (P< 0.01). HE staining indicated fistulous adenomas from small intestine and colon of two groups, Immunostaining showed Stat - 1 level in APC-min/+ tumors were increased and MSH-2 and vimentin-1 levels in APCmin/+ VDR-/- mice were increased, compared to APCmin/+tumors. Conclusion These observations suggested that inactivation of VDR promoted the intestinal tumor growth of APCmin/+ mice.

The Immunohistochemical Study of Oncogene and Tumor Suppressor Gene Proteins on Bone Tumor / 대한정형외과학회잡지

The discovery of oncogenes and tumor suppressor genes have made it possible to partly understand the mechanism of cancer development. It is generally accepted that the cancer development is caused by specific gene alterations and now more than 100 oncogenes and suppressor genes are known to be involved in human carcinogenesis. However, there are only a few reports about oncogene expression on bone tumors. The author carried out an immunohistochemical study to reveal the oncogene and suppressor genes on carcinogenesis of bone tumors using antibodies against c-myc, c-H-ras, p53 and EGF. In 32 cases of osteochondrorma, EGF, p53 and c-myc antisera revealed positive reaction in 4 (12.5%), 2 (6.3%) and 7 (21.9%) cases, and, in 4 cases of chondrosarcoma, c-myc antisera revealed positive reaction in 2 (50%) cases. In 21 cases of osteosarcoma, the positive reaction of p53 was noted in 10 (47.6%) cases and that of c-myc in 3 (14.3%) cases. In 14 cases of fibrous bone tumors, there are only 2 (14.3%) cases of positive reaction with p53. These results suggest some roles of the p53 and c-myc genes in osteosarcoma development and c-myc gene in osteochondroma and chondrosarcoma development.

Comparison of Ku protein Expression in CNE1 and CNE2 nasopharyngeal carcinoma cell lines / 中华放射肿瘤学杂志

Objective Radiosensitivity is highly correlated with DNA doub le stra nd breaks(DSBs). The repair of human DSB is possible mainly through nonhomologou s DNA end joining(NHEJ)pathway. This study was designed to determine the relat ionship between expression of 70Ku/ 80Ku prot ein (a modulating subunit of DNA-PK, which is an important component in NHEJ pathway) and radiosensitivity of nasoph aryngeal carcinoma cell lines. Methods Radiosensitivity parameters of the nasoph aryngeal carcinoma cell lines were obtained by the colony forming assay and radi ation dose-survival curve was done by linear quadratic model. Western blot was p erformed to determine the expression of 70Ku and 80 Ku in total extract of CNE1 a nd CNE2 at baseline level、different time after 4?Gy irradiation、12 h after di ff erent doses of irradiation. To perform a semi-quantitated assay of protein expr e ssion, the optic density (OD) value of each band was estimated by automatic imag e analysis system. Results Surviving fraction of CNE1 was higher than those of CNE2 at each dose point. Mean inactivation dose was higher in the CNE1(2.35)t han that in the CNE2(1.11), but the quantity of 70Ku/ 80Ku in either cell line was similar at both baseline and postirradiation levels. Conclusions CNE2 is mo re radiaosensitive than CNE1; as there was no marked correlation observed betwee n the quantity of Ku protein and radiosensitivity of the nasopharyngeal carcinom a cell lines. X-irradiation cannot result in any change in total quantity of Ku protein.