The aim of this paper was to investigate the immunosuppressive effects of dihydroartemisinin and Huobahua compatibility in mice with delayed hypersensitivity and explore its possible mechanism. The delayed-type hypersensitivity(DTH) model in mice was established to observe the immunosuppressive effects of dihydroartemisinin and Huobahua compatibility in DTH mice. ELISA assay was used to detect the contents of interferon(IFN-γ); histopathological changes and degree of mononuclear infiltration of right ear tissues were examined by HE staining; the expression level of intercellular cell adhesion molecule-1(ICAM-1) in the right ear of mice was detected by immunohistochemistry; the protein expression levels of p38 phospho mitogen activated protein kinase(p-p38 MAPK) was detected by Western blot analysis. As compared with the control group, the degree of ear swelling, thymus/spleen index, serum IFN-γ as well as the number and degree of infiltration of monocytes were significantly increased in the model group. As compared with the model group, the degree of ear swelling and thymus/spleen index of the mice in the combination group were significantly reduced; the number and degree of infiltration of monocytes were significantly relieved; the serum levels of IFN-γ and the expression levels of p-p38 MAPK and ICAM-1 proteins in the right ear were also significantly reduced. The combination of dihydroartemisinin and Huobahua can significantly inhibit the DTH response, and it may regulate the production and secretion of related inflammatory factor IFN-γ by inhibiting the phosphorylation activity of p38 MAPK, thereby further reducing the expression of ICAM-1 and thus exerting the immunosuppressive effect.
Animals ; Artemisinins ; Intercellular Adhesion Molecule-1/genetics* ; Mice ; Monocytes ; p38 Mitogen-Activated Protein Kinases/genetics*

OBJECTIVETo study the linkage between K469E polymorphism of intercellular adhesion molecule 1(ICAM1) gene with ICAM1 plasma level and coronary heart disease (CHD) in Han population of China.

METHODSOne hundred and sixty-four controls without CHD and 160 patients with CHD were enrolled in our study. By nested PCR with allele-specific oligonucleotide primers, all patients and controls were genotyped for the ICAM1 polymorphism. And the ICAM1 plasma level was measured by ELISA.

RESULTSIn the patients with CHD, both K allele frequency and the plasma level of ICAM1 were higher than those in control (P<0.05). The individual with K allele had higher plasma level of ICAM1 than that without K allele (344.34+/-128.59 microg/L vs 303.54+/-108.74 microg/L, P=0.008). K allele enhanced the risk of CHD (P<0.01, OR=2.158, 95%CI: 1.250-3.727). There was the K allele cooperation with smoking in influencing the risk of CHD.

CONCLUSIONThere is the polymorphism of ICAM1 K469E gene in Han population of China, and the K allele may be a genetic factor influencing the risk of CHD.

China ; ethnology ; Coronary Disease ; blood ; genetics ; Gene Frequency ; Humans ; Intercellular Adhesion Molecule-1 ; blood ; genetics ; Polymorphism, Genetic ; Potassium ; antagonists & inhibitors

OBJECTIVETo explore the effect of hemangiopoietin (HAPO) on the adhesive properties of human umbilical vein endothelial cells (HUVEC).

METHODSThe adhesion of HUVEC and the expressions of CD54, CD102, CD106, CD31, CD62E, and CD62P were measured by adhesion assay, flow cytometry, and semi-quantitative RT-PCR.

RESULTSHAPO enhanced the total adherence of HUVEC in a concentration-dependent manner. Flow cytometry analysis revealed that the treatment of HAPO resulted in a significantly increased expression of CD106 and CD62E on HUVEC in a time-dependent manner. When HUVEC were incubated with HAPO for 6 h, the percentage of CD106 + HUVEC and CD62E HUVEC increased about 2.10 folds and 5.84 folds, respectively, compared with control. The time-course of adhesive molecules mRNA expression indicated that the expression of CD106 and CD62E reached at the maximum 1.86 folds and 6.16 folds, respectively, compared with control.

CONCLUSIONHAPO may facilitate the homing of hematopoietic stem/progenitor cells.

Cell Adhesion Molecules ; metabolism ; Cells, Cultured ; E-Selectin ; biosynthesis ; genetics ; Endothelial Cells ; cytology ; Humans ; Intercellular Adhesion Molecule-1 ; biosynthesis ; genetics ; Proteoglycans ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Umbilical Veins ; cytology ; Vascular Cell Adhesion Molecule-1 ; biosynthesis ; genetics

OBJECTIVETo explore the effects of ICAM-1 gene transfection on the differentiation of MSCs to adipocytes.

METHODSThe recombinant retroviral expression plasmid MIGR1-ICAM-1 containing full length of mouse ICAM-1 gene was constructed. The constructed plasmid MIGR1-ICAM-1, empty plasmid MIGR1 and packaging plasmid ECOS were transfected into T293 cell lines and then the supernatant generated from T293 cells were used to infect mouse MSCs cell line C3H10T 1/2. The transfective efficiency was determined by inverted fluorescence microscope, real-time PCR and flow cytometry. Furthermore, ICAM-1 overexpressing MSCs (C3H10T 1/2-ICAM-1) and empty vector transfection MSCs (C3H10T 1/2-MIGR1) were cultured in medium with or without induction reagents, Oil-red-O staining was used to detect the lipid accumulation, and the expression of transcriptional factors C/EBPα and PPARγ, which were key factors in the differentiation of MSCs to adipocytes, were tested by real-time-PCR.

RESULTSThe recombinant retrovirus vector containing mouse ICAM-1 gene was successful constructed. After transfection into MSCs cell line C3H10T 1/2, the overexpression ICAM-1 MSCs cell line (C3H10T 1/2-ICAM-1) and control cell line (C3H10T 1/2-MIGR1) were obtained. Furthermore, these two cell lines were treated without or with adipocytic induction reagents, C3H10T 1/2-ICAM-1 showed significantly lower mRNA expression level for C/EBPα [(1.2 ± 0.7), (2.9 ± 0.9)] and PPARγ [(1557.6 ± 70.2), (7547.0 ± 442.2)] when compared with C3H10T 1/2-MIGR1 [(5.8 ± 0.5), (23.0 ± 2.3) and (2453.0 ± 215.6), (9856.3 ± 542.2)](P < 0.05). Moreover, little lipid droplet and decreased quantity of adipocytes were detected in C3H10T 1/2-ICAM-1 [(3.2 ± 0.5)/well, (12.2 ± 3.8)/well] than that in C3H10T 1/2-MIGR1 [(11.2 ± 0.4)/well, (51.3 ± 2.8)/well] (P < 0.05).

CONCLUSIONOverexpression of ICAM-1 in MSCs can inhibit its adipocytic differentiation.

Adipocytes ; cytology ; Animals ; Cell Differentiation ; Cell Line ; Intercellular Adhesion Molecule-1 ; genetics ; Mesenchymal Stromal Cells ; cytology ; Mice ; Transfection

Animals ; E-Selectin ; biosynthesis ; genetics ; Graft Rejection ; immunology ; pathology ; Intercellular Adhesion Molecule-1 ; biosynthesis ; genetics ; Liver Transplantation ; immunology ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Inbred Lew ; Vascular Cell Adhesion Molecule-1 ; biosynthesis ; genetics

OBJECTIVETo investigate the distributions of genotypic and allelic frequencies of intercellular adhesion molecule-1 (ICAM-1) gene K469E and platelet endothelial cell adhesion molecule-1 (PECAM-1) gene C373G in patients with preeclampsia.

METHODSPolymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) and DNA sequencing were used for detecting ICAM-1 gene K469E and PECAM-1 gene C373G genotypes in 110 women with preeclampsia and 110 normotensive pregnant women in comparison with their clinical characteristics.

ESULTSThe distributions of observed and expected genotype frequencies were consistent with Hardy-Weinberg equilibrium. No significant differences were found in the genotype and allele frequencies of ICAM-1 gene K469E between the two groups (P>0.05), but the CC and the CG genotype frequencies of PECAM-1 gene C373G were significantly different between them (P<0.05). The relative risk for preeclampsia of CG genotype was 1.959 folds of that in CC genotype carriers (OR=1.959, 95%CI: 1.090-3.520, P=0.024), and this association still existed after adjustment for age, gravidity, parity and BMI in logistic regression models. The C373G allele frequencies showed no significant difference between the two groups (P>0.05).

CONCLUSIONSThe CG genotype of PECAM-1 gene C373G genetically predispose the carriers to preeclampsia, while ICAM-1gene K469E polymorphisms is not associated with preeclampsia.

Adult ; Case-Control Studies ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; Platelet Endothelial Cell Adhesion Molecule-1 ; genetics ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Pre-Eclampsia ; genetics ; Pregnancy ; Sequence Analysis, DNA

To study the in vitro anti-angiogenesis effect of three curcumin pigments (curcumin, demethoxycurcumin, bisdemethoxycurcumin). In the study, the inhibitory effect of the three curcumin pigments on proliferation of HUVEC cells induced by OX-LDL and the effect on migration of HUVEC cells were detected. The effect on neovascularization was observed by chorioallantoic membrane (CAM) test. The effect on cell adhesion factors ICAM-1 and VCAM-1 of HUVECs were tested by Real-time RT-PCR. It was found that the three curcumins could inhibit the proliferation of HUVEC cells induced by OX-LDL within the dosage range 4, 8, 16 mg x L(-1), with a dose-dependence. The proliferative effect of curcumins on HUVECs was greater than the other two derivatives (P < 0.01). All of the three curcumin pigments inhibited the migration of HUVEC cells and the angiogenesis of chick chorioallantoic membrane (CAM). The migration inhibition rate of curcumins at middle and high concentrations was greater than the other two (P < 0.01). All of the three curcumin could down-regulate the expression of VEGF and ICAM-1, and curcumins showed more obvious effect in down-regulating VEGF than demethoxycurcumin and bisdemethoxycurcumin(P < 0.01); Bisdemethoxycurcumin showed the most significant effect in down-regulating ICAM-1 (P < 0.01). All of the three showed no remarkable effect on expression of VCAM-1, and only bisdemethoxycurcumin showed the down-regulating effect (P < 0.05). According to the findings, all of the three curcumin pigments could resist angiogenesis by inhibiting proliferation and migration of endothelial cells and down-regulating the expression of VEGF and adhesion molecules ICAM-1.
Angiogenesis Inhibitors ; pharmacology ; Animals ; Cell Movement ; drug effects ; Cells, Cultured ; Chorioallantoic Membrane ; drug effects ; Curcumin ; analogs & derivatives ; pharmacology ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; RNA, Messenger ; analysis ; Vascular Cell Adhesion Molecule-1 ; genetics ; Vascular Endothelial Growth Factor A ; genetics

OBJECTIVETo study the gene expression of adhesion molecules in pulmonary and hepatic microvascular endothelial cells during sepsis in mice.

METHODSMale mice were subjected to cecal ligation and puncture (CLP) and microvascular endothelial cells in pulmonary and hepatic tissues were harvested at 3 hours (early sepsis) and 12 hours (late sepsis) after CLP, respectively. Gene expression of the adhesion molecules was assessed by reverse transcription polymerase chain reaction (RT-PCR). Simultaneously, the alterations of myeloperoxidase (MPO) activity in pulmonary and hepatic tissues were also examined.

RESULTSE-selectin mRNA levels markedly increased at 3 hours after CLP in both pulmonary and hepatic microvascular endothelial cells, then they returned to the normal level at 12 hours after CLP. Increases in intercellular adhesion molecule-1 (ICAM-1) mRNA levels were found at 3 hours after CLP in both pulmonary and hepatic microvascular endothelial cells, and these levels became higher at 12 hours after CLP. Adhesion molecule-1 (VCAM-1) mRNA expression of vascular cells also increased significantly at 3 hours and 12 hours after CLP in both pulmonary and hepatic microvascular endothelial cells. The level of VCAM-1 mRNA in hepatic microvascular endothelial cells was higher at 3 hours than that at 12 hours after CLP, while the level of VCAM-1 mRNA in pulmonary microvascular endothelial cells was higher at 12 hours than that at 3 hours after CLP. The MPO activity in pulmonary and hepatic tissues increased at 3 hours after CLP, compared with that of the sham group. They both declined significantly at 12 hours after CLP, but they were still higher than that of the sham group.

CONCLUSIONSThe up-regulation of the gene expression of adhesion molecules in pulmonary and hepatic microvascular endothelial cells is an important step for the migration and accumulation of leukocytes at the site of inflammation, which plays a critical role in organ damage during sepsis. And the contribution of the heterogeneity of endothelial cells in organs' vulnerability during sepsis is worth a further investigation.

Animals ; Endothelium, Vascular ; cytology ; Gene Expression ; Intercellular Adhesion Molecule-1 ; genetics ; Liver ; cytology ; Lung ; cytology ; Male ; Mice ; Peroxidase ; metabolism ; Sepsis ; metabolism ; Up-Regulation ; Vascular Cell Adhesion Molecule-1 ; genetics

This study was aimed to clone human intercellular adhesion molecule-1 (ICAM-1) gene, to transfect the constructed eukaryotic expression vector ICAM-1-GFP into CHO cells, as well as to detect ICAM-1-GFP expression in CHO cells binding with Molt-4 cells. ICAM-1 cDNA gene was amplified by RT-PCR and inserted in PMD(18)-T vector. Then ICAM-1 cDNA from pMD18-ICAM-1 vector was subcloned into eukaryotic expression vector pEGFP-C1 to construct recombinant ICAM-1-pEGFP-C1 vector. Restriction analysis and DNA sequencing were used to confirm the recombinant vector. After stable transfection of CHO-K1 cells with the recombinant vector, the expression and subcellular localization of ICAM-1-GFP were detected by RT-PCR, flow cytometry and fluorescence microscopy. The function of ICAM-1-GFP fusion protein was assessed by the binding of ICAM-1-GFP/CHO cells to Molt-4 cells. The results showed that 1622 bp full-length ICAM-1 cDNA obtained and was successfully ligated with pMD(18)-T-vector, subcloned to construct recombinant ICAM-1-pEGFP-C1 vector. Restriction analysis and DNA sequencing indicated that recombinant ICAM-1-GFP was successfully constructed and ICAM-1-GFP was expressed stably in CHO cells. ICAM-1-GFP expression was only observed in the cytoplasm of ICAM-1-GFP/CHO cells by fluorescence microscopy. The ICAM-1-GFP/CHO cells were bound to PMA-treated Molt-4 cells. The expression of MEM-148 was very weak in PMA-treated Molt-4 cells. It is concluded that the ICAM-1-GFP eukaryotic expression vector has been constructed successfully and expresses stably in CHO cells. PMA can increase the binding of Molt-4 cells to ICAM-1-GFP/CHO cells by inducing specialized form of ICAM-1 clustering.
Animals ; CHO Cells ; Cricetinae ; Cricetulus ; DNA, Complementary ; genetics ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; Recombinant Proteins ; genetics ; Transfection

OBJECTIVETo explore the expression of NF-kappaB and ICAM-1 in the gas explosion wounded lung of rats and the relationship.

METHODSDigoxin labeled NF-kappaB was used as probe. In situ hybridization was performed to detect the NF-kappaB mRNA. Immunohistochemistry was used to detect the expression of NF-kappaB and ICAM-1.

RESULTSThe levels of NF-kappaB mRNA, the expression of NF-kappaB and ICAM-1 in the wounded rats were significantly increased and reached their peak two hours after injury. Pathology of lung tissue showed that some crachea epithelium mucosae were desquamated; congestion, edema of trachea wall and infiltration of neutrophilic granulocytes were found; hemorrhage, edema and infiltration of lots of inflammatory cells were present in alveolus cells. Electron microscope showed that type I, especially type II alveolus epithelia had degeneration and desquamation.

CONCLUSIONThe injury of gas explosion can activate NF-kappaB, which has close correlation with the acute injury to lung.

Animals ; Blast Injuries ; metabolism ; pathology ; Explosions ; Intercellular Adhesion Molecule-1 ; biosynthesis ; genetics ; Lung ; metabolism ; pathology ; NF-kappa B ; biosynthesis ; genetics ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley