Preparation,stability and in vitro activity of egg yolk immunoglobulin Y against human Sucrase / 中国免疫学杂志

Objective:To prepare the egg yolk immunoglobulin Y ( IgY) against human Sucrase and study its stability,in vitro activity. Methods:Hy-line laying hens were immunized with human Sucrase protein,IgY was isolated and purified from egg yolks of im-munized hens using water dilution and salting out method. Indirect ELISA was used to evaluate the titer and stability of IgY. The purity and specificity of IgY were analysed by SDS-PAGE and Western blot respectively. The inhibitory effects of IgY on α-glucosidase was studied by PNPG method. Results:Indirect ELISA results showed IgY could be detected on the tenth day after the first immunization, and the peak titer of IgY was 1:12 800 after the 40th day of immunization. SDS-PAGE showed that the heavy chain and light chain of IgY were 65 kD and 25 kD respectively, and the IgY against human Sucrase could specifically recognize the protein of human Sucrase. The IgY maintained primary titer when it was kept between 29-69℃ for 15 min,and pH 4-7,37℃,4 h. The titer of IgY was maintained 50% after digestion by pepsin and trypsin respectively for 2 hours. IgY had a higher resistence to pepsin than trypsin after longer digestion time. IgY showed an inhibitory effect on α-glucosidase in concentration dependent manner. The half inhibitory concentration (IC50) was 0. 540 mg/ml. Conclusion:The IgY against human Sucrase has been successfully obtained,which established foundations for its study of Type 2 diabetes mellitus rat models in vivo.

Study on garlic oil combined with 5-FU induced apoptosis of adenoid cystic carcinoma cell line ACC-M / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To observe the effect of garlic oil combined with 5-FU induced apoptosis of adenoid cystic carcinoma cell line ACC-M.@*METHOD@#Human salivary in adenoid cystic carcinoma cell line AC-M was cultured, divided into the experimental group (5-FU group, garlic oil group, garlic oil + 5-FU group) and the control group, to observe the growth activity of tumor cells by MTT methods; to analyse the changes of cell cycle and apoptosis rate by flow cytometry.@*RESULT@#MTT experiments showed that 5-FU, garlic oil, garlic oil and 5-FU on ACC-M cells have inhibition in different concentration, with the increase of concentration and action time of the rise; Cell cycle analysis showed significant changes in flow cytometry. With the increase of concentration and the acting time, the G0/G1, phase of the cell ratio increased, S had no significant change, but G2/M phase cells decreased. Apoptosis rate display showed garlic oil combined with 5-FU induced apoptosis of ACC-M cells was significantly stronger than single group.@*CONCLUSION@#Garlic oil can effectively induce the apoptosis of adenoid cystic carcinoma cell line ACC-M. The effect of garlic oil combined with 5-FU on ACC-M cells was stronger than the garlic oil, 5-FU used alone.

Study on inhibitory effects of allicin in mucoepidermoid carcinoma cells / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To observe the anticancer mechanism of allicin by observing the inhibitory effect of allicin on human salivary gland carcinoma cell line MEC-1.@*METHOD@#Cell proliferation were measured by MTT assay at different doses and different hours. In the meantime, cell cycle was detected via flow cytometry after different dose incubation with different hours.@*RESULT@#MTT results showed that the inhibitory rates of MEC-1 proliferations were increased in a concentration-and time-dependent manner. Flow cytometry analysis showed percent-age of MEC-1 cells decreased in G0/G1 phase and increased in G2/M. But there was no evident change in S phase. The cells were mainly blocked in M phase, and the inhibitory effect of the allicin on MEC-1 cells increased with the increasing of concentration and time.@*CONCLUSION@#Allicin can inhibit the growth of MEC-1 cells in vitro dramatically.

A genetic transformation study of introducing wild Gastrodia elata DNA into Solanum tuberosum by soaking seedling method / 中草药

Objective Using the soaking seedling method to introduce wild Gastrodia elata DNA into potato plantlets and analyze gastrodin of transformed Solanum tuberosum.Methods After the tuber grown up,the solution of gastrodin was extracted from the potatoes which were transformed by wild G.(elata) DNA.The transformed plants were scaned by ultraviolet.PCR was used to analyze GAFP gene by SDS-PAGE.TLC was used to analyze the gastrodin of transformed S.tuberosum.Results(1)The 21 transformed plants had obvious absorption peak at 220 nm in 200 transformed S.tuberosunr.(2)The fragment of wild G.elata GAFP gene was found by PCR from 5 transformed plants.(3)The protein expressions in transgenic S.tuberosum were obviously different from the normal S.tuberosum;a band was detected in transgenic S.tuberosum and wild G.elata,but wasn't detected in normal S.tuberosum.(4)The expression of the gastrodin was detected by TLC from three transformed plants. Conclusion It is feasible to use the soaking seedling method to introduce exogenous DNA into plants.