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Objective To study whether Nεcarboxymethyl lysine(CML)can form a good molecular docking with the scavenger receptor CD36and induce a stable interaction.Methods The interaction between CML and CD36was studied by co-immunoprecipitation.The binding mode and affinity of CD36to CML were tested using AutoDock 4.2,iBabel and XQuartz-2.7.7software respectively. Results Co-immunoprecipitation showed that anti-CD36antibody magnetic bead could precipitate CD36from the total protein in RAW264.7cells and anti-CML could detect CD36 binding CML.CD36had a good molecular docking with CML,CD36and CML interacted stably with each other.The affinity of CML to 4Q4Bprotein structure of CD4extracellular domain was -29.62kJ/mol.ARG82,ASN71and THR70were the products of amino acid receptor interaction. Further docking analysis showed that CML could form 3interacting hydrogen bonds with 4Q4B,and the docking prediction inhibition constant was 6.92with a root mean square deviation of 2.54.Conclusion A good molecular docking between CML and 4Q4Bprotein structure of CD36extracellular domain can induce a stable interaction between CML and CD36.Hydrogen bonding is the main interaction mode.
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Objective@#To investigate the clinical value of the serum new molecular markers, soluble triggering receptor expressed on myeloid cells-1(sTREM-1)and soluble hemoglobin scavenger receptor(sCD163), in the diagnosis of sepsis in elderly patients with burns.@*Methods@#A total of 58 inpatients with burns from Jun 2017 to June 2018 were enrolled in the study.Patients were divided into three groups: the sepsis group(n=12), the localized infection group(n=21)and the non-infection group(n=29). The levels of sTREM-1 and sCD163 were determined by enzyme-linked immunosorbent assays(ELISAs). The clinical diagnostic value of sTREM-1 and sCD163 was assessed by receiver operating characteristic(ROC)curve analysis.@*Results@#There was a statistically significant difference in the levels of sTREM-1 and sCD163 at day 1 between the three groups(F=20.994 and 38.363, P<0.01). Serum levels of sTREM-1 and sCD163 were higher in the sepsis group than in the localized infection group and the non-infection group.Serum levels of sTREM-1 and sCD163 were higher in the localized infection group than in the non-infection group.Serum levels of sTREM-1 and sCD163 were lower at day 7 than those at day 1 in all groups(F=21.242 and 41.035, P<0.01). Serum sTREM-1 levels were positively correlated with serum sCD163 levels(r=0.609, P=0.000). The AUC of sTREM-1 and sCD163 for the diagnosis of sepsis was 0.880(95%CI: 0.816~0.926).@*Conclusions@#Serum levels of sTREM-1 and sCD163 are elevated with increasing degrees of infection.Monitoring serum sTREM-1 and sCD163 levels is helpful for the diagnosis of sepsis in elderly patients with burns.
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Objective To investigate the clinical value of the serum new molecular markers,soluble triggering receptor expressed on myeloid cells-1 (sTREM-1)and soluble hemoglobin scavenger receptor(sCD163),in the diagnosis of sepsis in elderly patients with burns.Methods A total of 58 inpatients with burns from Jun 2017 to June 2018 were enrolled in the study.Patients were divided into three groups:the sepsis group(n=12),the localized infection group(n=21)and the non-infection group (n=29).The levels of sTREM-1 and sCD163 were determined by enzyme-linked immunosorbent assays(ELISAs).The clinical diagnostic value of sTREM-1 and sCD163 was assessed by receiver operating characteristic(ROC)curve analysis.Results There was a statistically significant difference in thelevels of sTREM-1 and sCD163 at day 1 between the three groups(F =20.994 and 38.363,P＜0.01).Serum levels of sTREM-1 and sCD163 were higher in the sepsis group than in the localized infection group and the non-infection group.Serum levels of sTREM-1 and sCD163 were higher in the localized infection group than in the non-infection group.Serum levels of sTREM-1 and sCD163 were lower at day 7 than those at day 1 in all groups(F=21.242 and 41.035,P＜0.01).Serum sTREM-1 levels were positively correlated with serum sCD163 levels (r =0.609,P =0.000).The AUC of sTREM-1 and sCD163 for the diagnosis of sepsis was 0.880(95％CI:0.816～0.926).Conclusions Serum levels of sTREM-1 and sCD163 are elevated with increasing degrees of infection.Monitoring serum sTREM-1 and sCD163 levels is helpful for the diagnosis of sepsis in elderly patients with burns.
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AIM:To construct the recombinant eukaryotic expression plasmid pEGFP-C1-SR-A I for the high expression in 293T cells in order to identify functions of savenger receptor-A I(SR-A).METHODS:The primer was designed according to MSR1 cDNA and pEGFP-C1-SR-A I was constructed by standard molecular cloning technique and enzyme digestion.After sequencing,the plasmid was transfected into 293T cells by lipidosome method.The expression of scavenger receptor-A I was identified by RT-PCR and Western blotting.The foam cells were evaluated by the formation of lipid granules in the cells with oil red staining.Cell adhesion was analyzed by cell adhesion assay.RESULTS:24 h after transfection,SR-A I mRNA was highly expressed and the high level of the protein was detected.The ratio of foam cell formation was doubled,the efficacy of cell adhesion was enhanced two times compared to the control group and the empty vector group.CONCLUSION:The recombinant eukaryotic expression plasmid has been constructed successfully with enhancing the function of uptake ox-LDL and adhesion in 293T cells by overexpression of SR-A I.
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AIM: To investigate the changes of expression of SR-BI in phorbol 12-myristate 13-acetate(PMA) differentiated U937 cells.METHODS: U937 cells were cultured with 100 nmol/L PMA in order to differentiate the cells to macrophages.Immunocytochemical method,Western blotting analysis and reverse transcription polymerase chain reaction(RT-PCR) were used to detect SR-BI protein and mRNA during differentiation.RESULTS: Immunocytochemistry showed that after exposure of U937 cells to PMA for 24,48,72 hours,the values of SR-BI protein expression in U937 cells were 15.94?3.56,27.86?4.39 and 9.08?2.37,with the first two higher than that in undifferentiated cells(7.76?1.74,P0.05) increment in the expression of SR-BI protein compared with U937 monocytes.RT-PCR showed that relative SR-BI mRNA expression in different group was 0.112?0.006,0.235?0.014,0.344?0.140 and 0.138?0.010,respectively.CONCLUSION: SR-BI protein and mRNA were increased after differentiation,reached a peak at 48 hours,and decreased at 72 hours.High expression levels of SR-BI in U937 macrophages following PMA differentiation may be correlated with foam cell formation.
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AIM: To investigate serum lipid and the expression of SR-B1 in the livers of diabetic mice. METHODS: Ten normal diet, female C57BL/6J mice, fifteen high fat and sugar diet female C57BL/6J mice, five fed 8 weeks and ten fed 16 weeks were used in the experiment. Serum total cholesterol (TC), triglycerides (TG), high density lipoprotein cholesterol (HDL-C), fasting blood glucose (FBG), insulin (INS) and the expression of SR-B1 in the livers were measured. RESULTS: 1. In the high fat and sugar diet mice, serum TC and FBG at 16 weeks were significantly higher than that in normal diet mice (P
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AIM: To observe the relationship between UCP2 mRNA expression in white adipose tissue and diet-induced obesity in SR-A I/II gene knock-out(SR-AⅠ/Ⅱ-/-) mice.METHODS: Fluorescent quantitative RT-PCR was used to detect UCP2 mRNA expression in mice epididymal white adipose tissue.The cellular morphological changes were analyzed by using image analysis.Serum TG,TC and LDL-C concentrations were measured by enzymatic determination.RESULTS: After fed with high fat diet for 12 weeks,average body weight of SR-A I/II-/-mice was much higher than that of wild type(SR-A I/II+/+) control mice(P
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AIM: To observe activity of scavenger receptor of macrophage derived from monocytes, the change of inflammation factor in plasma (including CRP, sICAM-1, sVCAM-1) and the influence of atorvastatin on activity of scavenger receptor in patients with coronary heart disease. METHODS: 75 patients with coronary heart disease (CHD), whose plasma lipid levels were normal, were divided into three groups: stable angina pectoris, unstable angina pectoris and acute myocardia infarction. 29 healthy persons were served as control. The level of C-reactive protein (CRP), soluble intercellular adhesion molecule-1 (sICAM-1) and soluble vascular cell adhesion molecule-1 (sVCAM-1) in plasma were determined in all subjects. Monocytes in peripheral blood were dissevered and cultured to transform into macrophage. The influence of atorvastatin on activity of scavenger receptor in macrophage was observed. RESULTS: The level of CRP, sICAM-1, sVCAM-1 and activity of scavenger receptor of macrophage in acute myocardia infarction group were higher than that in stable angina pectoris, unstable angina pectoris and control. Atorvastatin lowered the activity of scavenger receptor of macrophages derived from monocytes in patients with coronary heart disease. Activity of scavenger receptor of macrophages derived from monocytes in patients with coronary heart disease was correlated positively with CRP, sICAM-1 and sVCAM-1. CONCLUSION: Activity of scavenger receptor may be taken as index for monitoring the degree of active vulnerable atherosclerosis plaque. Atorvastatin may inhibit activity of scavenger receptor in macrophages derived from monocytes in patients with coronary heart disease.