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Objective To study the protective effects and mechanism of aqueous extract of arctium lappa root on vas?cular endothelial cell injury in hypertensive rats. Methods The hypertensive rat model was induced by N-nitro-L-argi?nine. Rats were randomly divided into normal control group, model control group, positive control group (aptopril 15 mg/kg), low concentration of aqueous extract of arctium lappa root (0.5 g/kg), medium concentration of (1 g/kg) and high concentra?tion of (2 g/kg) groups. After six weeks of continuous intragastric administration, the systolic blood pressure levels at tail ar?tery were measured at 1, 4, 7, 10, 13, 16, 19, 22, 29, 36 and 42 d after treatment. And other indicators related to inflammato?ry factors were detected including C-reactive protein (CRP) and interleukin (IL)-6. The intercellular adhesion molecule-1 (ICAM-1) level was detected by taking samples of thoracic aorta. Results (1) The systolic blood pressure level at tail ar?tery was significantly lower in aqueous extract of arctium lappa root group than that of model control group ( P<0.05). (2) The aqueous extract of arctium lappa root can significantly improve the vascular endothelial cell injury, suppress vascular endo?thelial cell loss and blood cell adhesion, and cell proliferation with collagen fibers in muscle membrane. ( 3) The serum levels of IL-6, CRP and vascular endothelial ICAM-1 were significantly reduced in aqueous extract of arctium lappa root group than that of model control group (P<0.05). Conclusion Aqueous extract of arctium lappa root can significantly improve vascular endothelial cell injury in hypertensive rats. The mechanism may be related to the inhibition of inflammatory cyto?kines like IL-6, CRP and the expression of ICAM-1, and the improvement of chronic inflammatory response in vascular en?dothelium of hypertensive rats.
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objective To explore the correlation of single nucleotide polymorphism (SNP) frequency of adiponectin(APN)in patients with type Ⅱ diabetes(T2DM)and its complications,investigate whether the SNP is a risk factor of inheritance of T2DM,and to set up a highly efficient.accurate, economical and practical screening assay to detect the mutation of APN in clinical practice.Methods According to the diagnostic criteria of T2DM,patients with coronary heart disease(CHD),hypertension (HP),and diabetic nephropathy(NE)were recruited into this study.In simple,12DM group,T2DM-HP, T2DM-CHD.T2DM-NE and the control group.serum biochemistry items are measured.The technique of denaturing high-performance liquid chromatography(DHPLC)was used to detect SNPs of ANP gene.Results After all fragments amplified in the reglen of promoter of APN gene were compared with APN GeneID:9370 sequence recorded in GenBank,point mutation has been identified(-11377G/C).The frequency of genotypes of GG,GC,and CC are 5.16%,42.25%,52.58%and 3.4%.32.75%,63.85%,respectively in the groups of T2DM and control.The frequency of G allele was related to the incidence of T2DM,and is a risk factor of T2DM.The relative risk of GC to CC in developing T2DM is much high than that in the control group (OR=0.55).By comparing the clinical data of different groups of genotype in T2DM,it was observed that the genotype affected systolic blood pressure,BMI,abdominal circumference, and waist-buttock ratio(P=0.015).After optimizing the experimental conditions.it was found column temperature 6 0℃ was the best when using DHPLC technology to estimate SNP of APN gene.Conclusion SNP (-11377G/C) of APN gene G allele has a definite correlation with complications of hypertension in T2DM patients,and may contribute to the genetic risk for type 2 diabetes.
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Objective Detection of single nucleutide polymorphisms in CD31-563 codon by denaturing high-performance liquid chromatography(DHPLC). Methods The eighth exon fragments of CD31 code gene in chromosome 17q23 were amplified by PCR. The fragments were analysed with DHPLC and were sequenced and compared with the sequences available in National Center for Biotechnology Information ( NCBI) database. Results The length of the amplified fragments is 203bp. There are three kinds of pictures when the fragments are analysed by DHPLC, after comparing with direct sequencing, the pictures of G/G homozygous、A/A homozygous、G/A heterozygous are obviously different, thus the individuals with the three kinds of genetype can be distinguished accurately. In 74 healthy people studied,the gene frequencies of CD31-563S(AGC) is 0.514, the gene frequencies of CD31-563N( AAC) is 0. 486. Conclusion DHPLC can effectively, economically and accurately detect the single nucleutide polymorphisms in CD31 -563 codon.
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Objective:To develop specific anti-AML T cells in vitro,and to study their biological characteristics and functions.Methods:Peripheral blood or bone marrow mononuclear cells (MNCs) were isolated from 12 patients with AML,and co-cultured with cytokine combinations in 96 well plates to be induced into dendritic cells (DCs).Cell morphology was observed under an inverted microscope during the first week and immunophenotype was detected by flow cytometry at day 7.Cytokine combination was replaced with high dose IL-2 at day 7 to promote specific anti-AML T cells.T cell phenotype was detected after 4 to 5 weeks.Lactate dehydrogenase (LDH) release assay was used to determine T cell killing activity.Results:After being cultured with cytokine combinations,the typical dendritic appearance with delicate membrane projections was observed.CD80,CD86 and HLA-DR markers were significantly upregulated(P