ABSTRACT
Objective The expression of chemokine C-X-C motif ligand 13 (CXCL13) within liver in primary biliary cholangitis (PBC) patients is significantly increased, but its origin and mechanism is not clear yet.The study aimed to investigate the expression of CXCL13 in the liver of mice through establishing a mouse model of PBC.Methods C57BL/6 mice were randomly divided into experiment group (n=20) control group(n=10).The mice in the experimental group were intraperitoneally injected with polyriboinosinic polyribocytidylic acid (Poly I:C) while the mice in control group were injected with PBS of the same volume.The level of serum AMA was quantified by ELISA and intrahepatic inflammatory cells were assessed by HE staining.Kupffer cells, liver sinusoidal endothelial cells, and infiltrating lymphocytes in the liver of mice were collected by in situ perfusion enzyme digestion and magnetic bead separation methods.The transcriptional level of intrahepatic CXCL13 in liver tissues and cell subpopulations were detected by qPCR.Results The serum AMA titers of the mice in experiment group increased gradually with the prolonging of modeling time and the positive rates at the 4th, 8th, and 12th week after the first injection of Poly I:C were 5.9%, 52.9% and 76.5% respectively.While the serum AMA titers of the mice in control group were at a lower level through the modeling process, with only 2 mice presenting a little higher level above positive cutoff value at the 12th week.The results of HE staining in liver tissues of both groups showed that there were a great amount of intensely infiltrating inflammatory cells in the mice of experimental group while no inflammatory cell infiltration were found in the mice of control group.The separation purity of Kupffer cells and liver sinusoidal endothelial cells in the mice of experiment group tested by flow cytometry were 76%-80%, 68%-72% respectively.Compared with the CXCL13 mRNA level in Kupffer cells [2.34(0.22-8.64)], the expression levels in liver sinusoidal endothelial cells and infiltrating lymphocytes declined[0.27(0.03-1.64), 0.05(0-0.22), P<0.05].Conclusion The chemokine CXCL13 is predominantly produced by Kupffer cells in the liver of PBC mice established by Poly I:C injection.
ABSTRACT
In this research, Casuarina eguisetifolia Linn was used to verify the broadly suitability of DNA bar-codes in identification of Li-medicine plants and systematic development of species. The genomic DNA of 22 samples collected C. eguisetifolia and its adulterants were amplified by 4 pairs of primers respectively (ITS (inter-nal transcribed spacer), ITS2 (internal transcribed spacer 2), trnH-psbA , rbcL) and sequenced bi-directionally. Obtained sequences were assembled using CodonCode Aligner. The dates were analysised using MEGA5.1 in ac-cordance with the kimura 2-parameter (K2P) model. The neighbor-joining (NJ) phylogenetic trees were construct-ed. Our study demonstrated the efficacy of ITS/ITS2 to distinguish between C. eguisetifolia and other adulterants species at the molecular level. Comparative to the primer of trnH-psbA and rbcL, there was a obviously DNA gap. The NJ trees showed that the several species of Casuarina can be classified to same types to show a obvi-ously monophyly, which the nearest family was Guttiferae. Therefore, ITS/ITS2 regions can accurately distinguish the original plant of Li-medicine. The systematic evolution of Casuarina can be verified in the molecular level.
ABSTRACT
Objective The effect of the pesticides on sister-chromatid exchange and micronuclei frequency of garlic root tip cells was researched. Methods The garlic root tips were treated with 13.65 mg/L phoxim, 25 mg/L lambda-cyhalothrin respectrely.With the positive and negative control, the garlic root tips were treated with 2%CP and distilled water respectively. Sister-chromatid exchange(SCE) and micronucle(iMCN)frequencies were calculated. Results As for the SCE frequency,lambda-cyhalothrin was the same as the negative control, metaldehyde and phoxim were more than the negative control (P