ABSTRACT
Objective:To detect the inflammatory reaction of macrophages induced by lipoglycans of different genotypes of Mycobacterium tuberculosis ( Mtb) in vitro and to analyze the differences in lipoglycan virulence. Methods:Lipoglycans were extracted from Mtb of Beijing, T1 and MANU2 genotypes and H37Rv by Triton X-114 liquid phase method and the crude extracts of lipoglycans was used to stimulate RAW264.7 macrophages. Changes in cytokine and receptor expression and cell apoptosis were detected 24 h after stimulation. The virulence of lipoglycans from different genotypes of Mtb was analyzed and compared. One-way analysis of variance and Tukey′s multiple comparisons test were used to compare the differences in various indexes between groups. Results:The expression of IL-10 at mRNA level induced by lipoglycans from Mtb of Beijing, T1 and MANU2 genotypes and H37Rv was (0.94±0.24), (1.86±0.24), (1.90±0.24) and (2.55±0.75) times that of the control group. Moreover, IL-10 mRNA expression induced by lipoglycans from Mtb of Beijing genotype was significantly lower than that of H37Rv group ( P<0.05). After stimulating RAW264.7 cells with the crude extracts of lipoglycans, the proportions of living cells in H37Rv, Beijing genotype, T1 genotype and MANU2 genotype groups were (72.75±2.25)%, (60.99±0.13)%, (80.66±0.40)% and (79.06±1.19)%, and the total cell apoptosis ratios was (10.42±0.23)%, (8.30±0.03)%, (9.24±0.79)% and (8.04±0.48)%, respectively. The proportion of living cells in Beijing genotype group was the lowest ( P<0.05), and the proportions of living cells in T1 and MANU2 genotype groups were higher than that in H37Rv group ( P<0.05). There was no significant difference in cell apoptosis ratio among the groups ( P>0.05). Lipoglycan-induced cell death was increased in Beijing genotype group, and the lipoglycan from Beijing genotype Mtb was more virulent than those from Mtb of T1 and MANU2 genotypes. Conclusions:Lipoglycan from Mtb of Beijing genotype could induce a higher level of cell death in vitro. It was an antigen component with stronger virulence than those from Mtb of T1 and MANU2 genotypes.
ABSTRACT
With the guide of state syndrome differentiation theory,this article states a combination method of disease and syndrome differentiation for treating chronic hepatitis B.We think it is a good way to treat chronic hepatitis B from regulating liver,spleen and kidney,treating qi and blood and strengthening,clearing and dissolving simultaneously.Clinically,we use the methods of strengthening spleen and kidney,soothing fiver,activating qi flow and blood circulation,clearing away heat and dampness,and removing toxin; and adopt the medicines of astragalus,herbal artemisiae,mushrooms,meat from rong,radix scutellariae,radix puerariae,notoginseng,radix bupleuri,liquorice,etc.