ABSTRACT
Objective To investigate the effect of naringenin on the killing rate of natural killer (NK) cells and related mechanism by amplification of human peripheral blood mononuclear cells into NK cells in vitro and co-culture with hepatocellular carcinoma (HCC) CLC5 cells at a ratio of 1∶ 1. Methods A lymphocyte separation medium was used to isolate human peripheral blood mononuclear cells, which were induced with recombinant human interleukin-2 in vitro to culture NK cells. CCK-8 assay was used to measure the proliferation of HCC cells after human HCC cells were treated with naringenin (0, 3.125, 6.25, 12.5, 25, and 50 μmol/L) for 0, 24, and 48 hours, and after human NK cells were treated with different concentrations of naringenin for 24 hours, CCK-8 assay was used to measure the proliferation of NK cells. CellTiter-LumiTM was used to measure the killing rate of NK cells after the NK-HCC cell co-culture system at the ratio of 1∶ 1 was treated with naringenin for 24 hours. Quantitative real-time PCR was used to measure the gene expression of the activating receptor NKG2D in NK cells and NKG2D ligands in HCC cells. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t -test was used for further comparison between two groups. Results After being induced and cultured by recombinant human interleukin-2, NK cells were amplified to 82.33%±0.70% of human peripheral blood mononuclear cells. After naringenin treatment for 24 hours, there was no significant difference in the proliferation rate of HCC CLC5 cells between all mass concentration groups (all P > 0.05), and in the 25 and 50 μmol/L mass concentration groups, naringenin significantly promoted the proliferation of NK cells (both P 0.05); it significantly upregulated the expression of the NKG2D ligands such as ULBP1 and ULBP3 in HCC cells (all P < 0.001). Conclusion Naringenin may increase the killing activity of NK cells by upregulating the expression of NKG2D ligands in HCC cells.
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Objective To determine whether the antiserum produced by immunizing mice with conotoxin GI coupled with bovine serum albumin (BSA) could neutralize GI conotoxin.Methods The GI-BSA was prepared by glutaraldehyde-coupled method,and the mice were immunized with the GI-BSA to produce antiserum.The antibody neutralization assay was used to test the detoxication of the antiserum.Results The SDS-PAGE protein electrophoresis showed that the coupling reaction of GI hapten with BSA was successful.The two distinct protein bands of GI-BSA were more than 120×103.Each mouse was immunized four times with 99 μg every two weeks.After the fourth immunization,the serum neutralization titer was more than 1:64 000.After the intraperitoneal injection of the mixture of 100 or 200 μl of the antiserum and different doses of GI,75% of the mice survived in the group with 100 μl of the antiserum and 1× LD50 GI(16.3 μg/kg).The same percentage of mice also survived in the group of with 200 μl of serum and 25.8 μg/kg of GI.Conclusion The antiserum produced by immunizing mice with GI-BSA exhibits significant detoxication activity to conotoxin GI.
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Anti-microbial properties of 21 endophytic fungal strains from Hypericum perforatum Linn, were evaluated against three human pathogens, Staphyloccocus aureus, Escherichia coll and Rhodotorula glutinis, and two phytopathogens, Rhizoctonia cerealis and Pyricularia grisea
The results indicated that the ethyl acetate extracts of endophytic fermentation broth had stronger anti-microbial activities than their fermentation broth. And the inhibitory effect of the endophytic extracts on human pathogens was better than those on phytopathogens. Among these endophytic fungi, strains GYLQ-10, GYLQ-24 and GYLQ-22 respectively showed the strongest activities against S. aureu, E. coli, R. glutinis. GYLQ-14 and GYLQ-22 exhibited the most pronounced effect on P. Grisea while both GYLQ-06 and GYLQ-08 had the strongest anti-microbial activities against R. cerealis. Till now, this study is the first report on the isolation of endophytic fungi from H. Perforatum Linn, and their anti-microbial evaluation
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Background and purpose: Carcinosarcoma of the lung is a rare malignant pulmonary neoplasm,which including epithelial and parenchymal malignant structure with a poor prognosis. The purpose of this study was to describe the clinicopathologic characteristics and the survival of pulmonary carcinosarcoma. Methods: From Jan.1980 to Dec. 2006, 64 patients with pulmonary carcinosarcoma who underwent surgical treatment were retrospectively analyzed. Results: The overall 5-year survival rate of the patients was 14.1%. There was significant difference between stage Ⅰ/Ⅱ and stage Ⅲ/Ⅳ disease (28.6% vs 2.8%, P=0.003), respectively. The 5-year survival rates of lobectomy, pneumonectomy and palliative resection were 3 3.3 %, 2.8% and 0% ( P=0.003 ), respectively. Conclusion:p-TNM and operative pattern were correlated with survival. Early diagnosis and radical operation are important to the survival of the patients with pulmonary carcinosarcoma.