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Objective To investigate the effect of plasma membrane-associated sialidase 3(NEU3) activity on the proliferation and apoptosis of osteosarcoma MG-63 cells in vitro. Methods MG-63 cells were cultured in vitro. Anti-NEU3 antibody(Ab)immunofluorescent staining was used to indicate the cellular locali-zation of NEU3 in MG-63 cells. The cells treated with 0 nmol/ L 2-deoxy-2,3-didehydro-N-acetyl neuraminic acid(DANA)or 0 μg/ ml anti-NEU3 Ab were used as blank control groups. The cells were treated with 10, 20,50 nmol/ L DANA,or 0. 5,1. 0,2. 0 μg/ ml anti-NEU3 Ab for 24 h or 48 h,respectively. The inhibition rates of the cell proliferation and cell apoptosis rates were measured with CCK-8 and flow cytometry. The expression levels of oncogene-related proteins,Ras protein and Bcl-2 protein,were detected by Western blotting. Results The immunofluorescence result showed that NEU3 was located in the cytoplasm of MG-63 cell. After treating with 0,10,20,50 nmol/ L DANA for 48 h,the inhibition rates of cell proliferation were 0, 15. 10% ± 3. 23% ,41. 46% ± 2. 31% ,64. 68% ± 4. 12% ,with significant statistical difference(F = 99. 90, P < 0. 001),and the following contrast between each two groups met the statistical significance(all P < 0. 05). After treating with 0,0. 5,1. 0,2. 0 μg/ ml anti-NEU3 Ab for 48 h,the inhibition rates of cell proliferation were 0,9. 34% ± 1. 53% ,19. 66% ± 4. 18% ,42. 50% ± 5. 68% ,and the difference was statistically signifi-cant(F = 25. 67,P < 0. 001),and the following contrast between each two groups met the statistical signifi-cance(P < 0. 05),except the difference between 0. 5 and 1. 0 μg/ ml groups(P > 0. 05). When the MG-63 cells were treated with 0,10,20,50 nmol/ L DANA for 24 h,the cell apoptosis rates were 4. 05% ± 0. 07% , 4. 15% ± 0. 23% ,12. 85% ± 1. 48% ,8. 29% ± 0. 86% ,respectively,and the difference was statistically sig-nificant(F = 23. 21,P < 0. 001). And the following contrast between each two groups met the statistical signi-ficance(P < 0. 05),except the differences between 0 nmol/ L and 10 nmol/ L,20 nmol/ L and 50 nmol/ L groups(P > 0. 05). When the MG-63 cells were treated with 0,0. 5,1. 0,2. 0 μg/ ml anti-NEU3 Ab for 24 h,the cell apoptosis rates were 4. 05% ± 0. 07% ,20. 13% ± 2. 97% ,20. 29% ± 2. 82% ,20. 58% ± 0. 70% ,with statistical significant difference(F = 15. 36,P = 0. 001). And the following contrast between each two groups showed that the differences between 0 μg/ ml and each treated group were statistically signifi-cant(P < 0. 05),while the differences between two treated groups were not statistically significant( P >0. 05). Western blotting results showed that the expression levels of Ras and Bcl-2 decreased with the increasing concentrations of DANA and anti-NEU3. Conclusion Inhibition of NEU3 enzyme activity can suppress the survival rate of MG63 cells and increase the cell apoptosis. The possible mechanism may be related to the declined expression of oncogene-related proteins Ras and Bcl-2,which suggests that NEU3 may be a possible target for treating osteosarcoma.
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Objective To probe the diagnostic value of MRI in synovial hemangioma of the knee.Methods Seven patients(2 male,5 female,age ranged from 8 to 58 years with mean age 25.1 years) selected from 2025 cases with MRI examination of knee,which were proved to be intra-articular hemangiomas histologically or clinically.All of the patients had plain film before MRI examination.MR findings of synovial hemangioma of knee were analysed ultrasound.Results Four hemangiomas occurred in infrapatellar bursa,others 3 cases in both infrapatellar and suprapatellar bursa were involved.MR signal intensity of hemangiomas appeared three types:(1)nodular hypointensity on both T_1WI and T_2WI in 2 cases;(2)iso-hypointensity on T_1WI and hyper-hypointensity on T_2WI in 2 cases;(3)hypointensity mixed with isontensity on T_1WI and nodular of linear mixedintensity on T_2WI in 3 cases.The difference of synovial hemangioma of knee in histologic elements,the MR findings were different.Conclusion MRI features of synovial hemangioma are correlated with pathology histologic elements.
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Aim To observe the effect of recombinant human interferon and verazole used alone or in combination in resisting respiratory synthesis virus (RSV) in vitro. Methods RSV strains were proliferated with Hela cells in Eagles solution on a 96 hole plate. The recombinant human interferon and virazole were diluted to different concentrations and were separately added in the dose of 100 ?l to each hole of the plate. After 48 hours cultured, the concentrations of the drugs for inhibiting cytopathogenic effect(CPE)of RSV were determined. Results When the concentration of interferon was ≥5 U?ml -1 and virazlole ≥24 ?g?ml -1 ,respectively,the effect of the two drugs on inhibiting the CPE of RSV was remarkable and was improved with their concentration increasing .When the concentrations of the two drugs were lower than that of their effect respectively , their united use also had obvious effect in resisting the virus. In addition, the different using methods of interferon have also different results. Conclusion Both recombinant human interferon and virazole are effective in inhibiting RSV in vitro and will bring about better effect when used in combination.
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AIM: To study the possibility and conditions of transplacental infection of coxsackievirus B3(CVB 3) from pregnant mice to their fetuses and newborns. METHODS: Coxsackievirus B 3 strain causing balb/c mice myocardial injury(CVB 3m )was inoculated with 10 5 TCID 50 in dose into the mother mice at 6-7 days (early gestation),9-10 days (middle gestation) and 17-18 days (late gestation) of gestation, in contrast with non pregnant mice. Some placentas and fetuses were removed by caesarean section before mothers partusing; some mothers and their babies were sacrificed after parturition, and virus isolation, serological and pathological tests were performed. RESULTS: Viramiae was observed in mother mice of late gestation inoculated with CVB 3m at a fit amount on the second day after inoculation, while no newtralizing antibody to CVB 3m was detected in blood. The virus was isolated from cardiac muscles of inoculated mother mice in different gestation and the controls. The virus was also isolated from some placentas and fetuses, and both sera and cardiac muscles of infants in the late gestation (virus titer were all 10 -2 -10 -3 ). On d 7 of inoculating virus, pregnant and non pregnant mice titers of neutralizing antibody to CVB 3m in sera were all between 1160 and 1320. Under the electromicroscopy, some cardiac muscle cells of mother or infant mice appeared with morphological changes and little hollow bubbles occured in cytoplasm. The fibers broke off, and the bright and dark belts became indistinct. CONCLUSION: The amimal model, intraplacental passage of CVB 3 from pregnant mother in late gestation to fetus in mice, is a benefitial tool to study enterovirus diseases in human perinatal period.