OBJECTIVE: Growth-arrest-specific protein 6 (Gas6) is a vitamin K-dependent protein and involved in cell proliferation, survival, adhesion and migration . Also it has been shown to play an important role in the inflammatory response .The aim of present study was to investigate the role of Gas6 in the process of the expression of adhesion molecules and chemokines of human umbilical vein endothelial cells (HUVECs) induced by Porphyromonas gingivalis lipopolysaccharide(P.g-LPS). METHODS: After up-regulation and down-regulation of the expression of Gas6, the vascular endothelial cells were stimulated with 1 mg/L P.g-LPS for 3 h and 24 h. Real-time quantitative polymerase chain reaction(real-time PCR) was taken to detect the expression of the cell adhesion molecules:intercellular adhesion molecule-1 (ICAM-1) and E-selectin, as well as chemokines:interleukin-8 (IL-8) and monocyte chemoattractant protein 1 (MCP-1). Wound healing assay was taken to observe the migration ability of endothelium cells in different groups. RESULTS: After 3 h of P.g-LPS stimulation, the expression of adhesion molecules and chemokine in the down-regulation group was not significantly different from that in the control group,while in the up-regulation group the decrease of E-selectin, ICAM-1, IL-8 and MCP-1 was 81%±0%, 47%±3%, 76% ± 3%, 26% ± 6% respectively. After 24 h of P.g-LPS stimulation, the expression of adhesion molecules and chemokine in down-regulation group was significantly higher than that in control group (2.06±0.07, 1.99±0.11, 3.14±0.15, 1.84±0.03 flod), while these molecules in the down-regulation group was significantly lower than in the control group (29%±1%, 62%±3%, 69%±1%, 41%±2%). Differences were statistically significant (P<0.01). Wounding healing assay showed that down-regulation of Gas6 enhanced migration ability of endothelial cells while up-regulation of Gas6 weakened this ability,which was consistent with the trend of real-time PCR result. CONCLUSION Down-regulation of the Gas6 gene enhanced the expression of ICAM-1, E-selectin, IL-8 and MCP-1 in HUVECs after P.g- LPS stimulating, while up-regulaiton of the Gas6 gene weakened the expression of ICAM-1, E-selectin, IL-8 and MCP-1 in HUVECs after P.g-LPS stimulating,suggesting that Gas6 may play a role in the process of endothelial cell adhesion.
Cell Adhesion ; Cell Adhesion Molecule-1 ; Cells, Cultured ; Chemokines/metabolism* ; E-Selectin/metabolism* ; Endothelium, Vascular ; Humans ; Intercellular Signaling Peptides and Proteins/physiology* ; Lipopolysaccharides ; Porphyromonas gingivalis/metabolism* ; Vascular Cell Adhesion Molecule-1/metabolism* ; Vitamin K

Cell Hypoxia ; physiology ; Cells, Cultured ; Diabetic Retinopathy ; metabolism ; Female ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; Male ; Middle Aged ; Monocytes ; metabolism ; Prospective Studies ; Vascular Endothelial Growth Factors ; metabolism

OBJECTIVETo explore the effect of drug-containing serum of Chinese herbal compounds [Xiongshao Capsule (XS, for activating blood) and Huanglian Capsule (HL, for dispelling toxin)] on tumor necrosis factor-alpha (TNF-alpha)-induced adherence between human umbilical vein endothelial cells (HUVECs) and polymorphonuclear neutrophils (PMN), inflammatory reaction and expression of related proteins in mitogen-activated protein kinase (MAPK) pathway.

METHODSThirty-two rats were randomly divided into four groups (8 in each group) using random digit table: the blank control group treated with distilled water, the test group I treated with Chinese herbal compound of XS (0.135 g/kg), the test group II treated with Chinese herbal compound of HL (0.135 g/kg), and the test group Ill treated with Chinese herbal compound of XS (0.135 g/kg) and HL (0.135 g/kg). All medication was given by gastrogavage once a day for a week. Rats' blood serum was harvested 1 h after the last administration to prepare drug-containing serum. HUVECs were exposed to TNF-alpha (100 ng/mL) to induce cell injury model and incubated with corresponding drug-containing serum (10%) for 24 h. Normal rats' serum was given to cells in the blank control group and the model group, while XC + HL containing serum was given to cells in the rest 3 groups. The adherence of HUVECs and PMN cells was detected by using rose bengal strain. Levels of E-selectin, intercellular adhesion molecule-1 (ICAM-1), and interleukin-1beta (IL-1P) in the supernatant of cultured HU-VECs were determined by ELISA. Protein expressions of mitogen-activated protein kinases p38 (p38MAPK) and extracellular signal-regulated kinase 1/2 (ERK 12) were determined by Western blot.

RESULTSCompared with the blank control group, HUVECs were seriously injured; PMN adherence amount significantly increased; levels of E-selectin, ICAM-1, and IL-1beta increased; expression levels of p-p38MAPK and p-ERK 1/2 in the supernatant of HUVECs significantly increased in the model group (all P < 0.01). Compared with the model group, HUVECs-PMN adherence amount decreased (P < 0.05); levels of E-selectin, ICAM-1, and IL-1 beta in the supernatant of HUVECs decreased (P < 0.01, P < 0.05); expression levels of p-p38MAPK and p-ERK 1/2 of endothelial cells decreased in the test group I, II, and III (P < 0.01).

CONCLUSIONSDrug-containing serums of activating blood, activating blood and dispelling toxin could attenuate TNF-alpha induced injury of HUVECs, inhibit HUVECs-PMN adherence and the release of adhesion factors. Its mechanism might be involved with protein phosphorylation of p38MAPK and ERK 1/2 in the MAPK pathway.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; E-Selectin ; Endothelial Cells ; physiology ; Human Umbilical Vein Endothelial Cells ; Humans ; Inflammation ; Intercellular Adhesion Molecule-1 ; metabolism ; Interleukin-1beta ; Mitogen-Activated Protein Kinase 3 ; Neutrophils ; Rats ; Serum ; Tumor Necrosis Factor-alpha ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

BACKGROUNDTotal knee arthroplasty (TKA) is a successful and frequently performed procedure in orthopedic surgery. The diagnosis of peri-prosthetic joint infection following TKA remains challenging. The present study estimated the usefulness of knee skin temperature (measured by infrared thermography) and serum soluble intercellular adhesion molecule-1 (sICAM-1) in the diagnosis of post-operative knee peri-prosthetic infection.

METHODSPatients were divided into three groups: 21 patients undergoing uncomplicated TKAs, seven with prosthesis infection, and three undergoing TKA revisions. The serum levels of interleukin-6 (IL-6), C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), and sICAM-1 as well as the local knee skin temperature were measured pre-operatively and on Days 1 and 7 and at 1, 3, and 6 months post-operatively in Groups 1 and 3. The same parameters were measured in Group 2 at the time of prosthesis infection diagnosis.

RESULTSIn Group 1, the levels of IL-6, CRP, ESR, and knee skin temperature were significantly elevated post-operatively, but returned to baseline levels within 6 months. The sICAM-1 levels were not significantly different. The mean differential temperature (MDT) and levels of siCAM-1, IL-6, CRP, and ESR differed significantly between Groups 1 and 2. The MDT had returned to normal in Group 3 by 6 months post-operatively.

CONCLUSIONSElevations in IL-6, CRP, ESR, and MDT in patients undergoing TKA could be a normal response to surgical trauma, but sustained elevations may be indicative of complications. The knee skin temperature and sICAM-1 may be used as indicators in the diagnosis of knee prosthesis infection following TKA.

Adult ; Aged ; Arthroplasty, Replacement, Knee ; adverse effects ; Blood Sedimentation ; C-Reactive Protein ; metabolism ; Female ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; Interleukin-6 ; blood ; Knee Joint ; immunology ; surgery ; Male ; Middle Aged ; Prospective Studies ; Skin Temperature ; physiology ; Thermography ; methods

BACKGROUNDPoly(ADP-ribose) polymerase (PARP) plays an important role in the death of retinal capillary cells in diabetic retinopathy (DR) partly via its regulation of nuclear factor kappa B (NF-κB). The current study investigated the effect of the regimen of Gaoshan Hongjingtian (RG) on the mechanism of PARP regulation of NF-κB, and demonstrated the possible impact of the RG and Gaoshan Hongjingtian (Rhodiola sachalinensis, RS) on diabetic retinopathy.

METHODSWistar rats were made diabetic by administering streptozotocin. They were then assigned to three groups at random. After 2 months, the three groups of these diabetic rats were treated with RS or RG, or untreated. Analyses of expression levels of PARP, NF-κB, and intercellular adhesion molecule-1 (ICAM-1) in the retinas of rats in different groups were performed by Western blotting and immunohistochemical assays, and mRNA levels of NF-κB and ICAM-1 were determined by real-time polymerase chain reaction (PCR). In addition, the basement membranes of capillaries in the rats' retinas were observed using electron microscopy, and diabetes-induced capillary degeneration (ghost pericytes and acellular capillaries) were quantitated.

RESULTSFrom the third month after the injection of streptozotocin, the diabetic rats were given daily RG, RS or tap water separately. The diabetic rats failed to gain weight compared with normal age-matched rats, whereas their glycated hemoglobin levels were significantly increased. After 5 months, the mRNA levels of NF-κB and ICAM-1 and the protein expression of PARP, NF-κB, and ICAM-1 were significantly increased in the retinas of diabetic rats in the untreated group compared with the nondiabetic controls. After 8 months, the number of degenerated retinal capillaries (ghost pericytes and acellular capillaries) was significantly increased in the diabetic rats in the untreated group compared with normal age-matched rats. RG and RS inhibited diabetes-induced over-expression of PARP, NF-κB, and ICAM-1 in the retinas of diabetic rats at the end of 5-month diabetic duration. Treatment using RG and RS significantly inhibited increases in the number of acellular capillaries and pericyte ghosts and suppressed the basement membrane thickening in the retinas of rats with diabetes for 8 months compared with the control diabetic rats.

CONCLUSIONSThese results indicate that PARP plays an important role in the pathogenesis of diabetic retinopathy. RS and RG may have acted on the mechanism of PARP regulation of NF-κB, which suppressed the expression of NF-κB and ICAM-1, and led to the inhibition of retinal capillary degeneration.

Animals ; Basement Membrane ; drug effects ; pathology ; Diabetes Mellitus, Experimental ; drug therapy ; pathology ; Diabetic Retinopathy ; drug therapy ; pathology ; Intercellular Adhesion Molecule-1 ; genetics ; Male ; Medicine, Chinese Traditional ; NF-kappa B ; genetics ; physiology ; Poly(ADP-ribose) Polymerases ; physiology ; Rats ; Rats, Wistar ; Rhodiola ; Streptozocin

OBJECTIVETo investigate the changes of adhesion molecules, cyclic adenosine monophosphate(cAMP) and cyclic guanosine monophosphate (cGMP) in divers post 480 heliox saturation diving.

METHODSFour divers were compressed within 96 hours to depths of 480 m with heliox-oxygen and held at the designated depth for 49 hours, excursion to 493 m during their saturation stay, then decompressed within 302 hours to the surface. The blood samples were collected before compression and post decompression, the expression level of intercellular adhesion molecule-1(ICAM-1), E-selectin, P-selectin, cAMP, cGMP were detected with ELISA analysis box.

RESULTSCompared with the levels of CAMs before compression, the levels of ICAM-1, E-selectin, P-selectin and cGMP in the serum were changed post decompression (P > 0.05). The levels of cAMP were significantly elevated post decompression (629.91 +/- 75.01) nmol/L vs (66.72 +/- 83.15) (P < 0.05).

CONCLUSIONThe decompression schedule in this heliox saturation diving is safe, the decompression sickness pathology in this diving has not been induced. But the stress response of divers are enhanced by this great depth saturation diving.

Cyclic AMP ; blood ; Diving ; physiology ; E-Selectin ; blood ; Helium ; Humans ; Intercellular Adhesion Molecule-1 ; blood ; Oxygen ; P-Selectin ; blood

OBJECTIVETo extract tumor interstitial fluid (TIF) from MKN-45 gastric cancer which is similar to "muddy phlegm" in Chinese medicine and observe influences of MKN-45 tumor interstitial fluid (MKN-45 TIF) intervention on metastasis of gastric cancer and on the expressions of vascular endothelial growth factor (VEGF), kinase insert domain containing receptor (KDR), epithelial-cadherin (E-cad), cyclooxygenase-2 (COX-2), intercellular adhesion molecule-1 (ICAM-1) and telomerase genes and proteins in primary tumor tissue.

METHODSAn MKN-45 tumor-bearing model was established in 50 nude mice. The modeled animals were equally randomized to 5 groups: the simple tumor-bearing group (model group), the normal saline (NS) via tail vein injection (i.v.) group (NS i.v. group), MKN-45 TIF i.v. group (TIF i.v. group), NS intraperitoneal injection (i.p.) group (NS i.p. group), and MKN-45 TIF i.p. group (TIF i.p. group). The TIF and NS intervention groups received injection (i.p. or i.v.) of MKN-45 TIF or NS twice a week, 0.2 mL at a time. After 8 weeks, the primary tumors were removed, weighed and HE stained to observe tumor metastasis. The primary tumor tissues were analyzed by immunohistochemistry and real-time quantitative PCR to detect expressions of VEGF, KDR, E-cad, COX-2, ICAM-1, and telomerase genes and proteins in different groups.

RESULTSThere were significant differences in tumor weight between TIF intervention groups and the model and NS intervention groups. Tumor metastasis was observed in all 5 groups, but the tumor metastasis rate in TIF intervention groups was significantly higher than those in the model and NS intervention groups. The gene and protein expressions of gastric cancer-related factors VEGF, KDR, COX-2, ICAM-1 and telomerase were unregulated while the gene and protein expressions of E-cad were downregulated in TIF intervention groups.

CONCLUSIONSTIF promotes tumor growth, invasion and metastasis of gastric cancer. These findings provide preliminary experimental clues for verifying the hypothesis of "tumor-phlegm microenvironment".

Animals ; Cadherins ; genetics ; metabolism ; Cell Line, Tumor ; Cyclooxygenase 2 ; genetics ; metabolism ; Extracellular Fluid ; metabolism ; Gene Expression Regulation, Neoplastic ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; metabolism ; Male ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Stomach Neoplasms ; metabolism ; secondary ; Telomerase ; genetics ; metabolism ; Tumor Microenvironment ; physiology ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; metabolism

The purpose of this study is to determine 1) whether morphine postconditiong (MPostC) can attenuate the intercellular adhesion molecules-1 (ICAM-1) expression after reoxygenation injury and 2) the subtype(s) of the opioid receptors (ORs) that are involved with MPostC. Human umbilical vein endothelial cells (HUVECs) were subjected to 6 hr anoxia followed by 12 hr reoxygenation. Three morphine concentrations (0.3, 3, 30 microM) were used to evaluate the protective effect of MPostC. We also investigated blockading the OR subtypes' effects on MPostC by using three antagonists (a micro-OR antagonist naloxone, a kappa-OR antagonist nor-binaltorphimine, and a delta-OR antagonist naltrindole) and the inhibitor of protein kinase C (PKC) chelerythrine. As results, the ICAM-1 expression was significantly reduced in the MPostC (3, 30 microM) groups compared to the control group at 1, 6, 9, and 12 hours reoxygenation time. As a consequence, neutrophil adhesion was also decreased after MPostC. These effects were abolished by coadministering chelerythrine, nor-binaltorphimine or naltrindole, but not with naloxone. In conclusion, it is assumed that MPostC could attenuate the expression of ICAM-1 on endothelial cells during reoxygenation via the kappa and delta-OR (opioid receptor)-specific pathway, and this also involves a PKC-dependent pathway.
Animals ; Benzophenanthridines/pharmacology ; Endothelial Cells/cytology/*drug effects/*metabolism ; Endothelium, Vascular/cytology ; Humans ; Intercellular Adhesion Molecule-1/genetics/*metabolism ; Morphine/*pharmacology ; Naloxone/pharmacology ; Naltrexone/analogs & derivatives/pharmacology ; Narcotic Antagonists/pharmacology ; Narcotics/*pharmacology ; Protein Isoforms/metabolism ; Protein Kinase C/antagonists & inhibitors/metabolism ; Receptors, Opioid/metabolism ; Reperfusion Injury/*metabolism ; Signal Transduction/physiology ; Umbilical Veins/cytology

OBJECTIVETo investigate the interference effects of lymph plasma on endotoxic shock and its mechanism.

METHODSSixty Wistar male rats were randomly divided into three groups: control group, model group and lymph plasma group. The endotoxic shock model of rats were duplicated by jugular intravenous injection with LPS (15 mg/kg), and after 15 minutes the normal lymph plasma was infused in lymph plasma group rats, which amount of lymph plasma was one fifteenth of whole blood volume. The effects of lymph plasma on the mean arterial pressure (MAP), microcirculation of mesentery near the ileum lower, the numbers of leukocytes adherent to the venular wall, the P-selectin and intercellular adhesion molecule-1 (ICAM-1) in plasma were observed.

RESULTSThe normal lymph plasma might be prevent from progressive decreased of MAP, depress the pathologic diameter constriction of mesenteric microvessels, reduce leukocytes adherent to the venular wall, improve blood flow condition of microcirculation, and decrease the level of P-selectin and ICAM-1 in plasma (P < 0.05, P < 0.01).

CONCLUSIONA little of normal lymph plasma plays a positive interference effect on microcirculatory dysfunction and hypotension with endotoxic shock induced by LPS challenge. Those mechanisms may be concerned with decreasing the production of cell adhesion molecules.

Animals ; Intercellular Adhesion Molecule-1 ; blood ; Lipopolysaccharides ; Lymph ; physiology ; Male ; Mesentery ; blood supply ; Microcirculation ; physiology ; P-Selectin ; blood ; Random Allocation ; Rats ; Rats, Wistar ; Shock, Septic ; chemically induced ; physiopathology

OBJECTIVETo explore the role of intercellular adhesion molecule-1 (ICAM-1) and regulated upon activation normal T cell expressed and secreted (RANTES) in bronchiolitis and their correlation in the pathogenesis of this disorder.

METHODSThe expression of ICAM-1 was detected by flow cytometry on lymphocytes of peripheral blood in 28 infants with bronchiolitis, 23 infants with bronchopneumonia and 24 healthy infants. Serum level of RANTES was assayed using ELISA. The correlation between ICAM-1 and RANTES levels was evaluated using Pearson correlation coefficient.

RESULTSThe ICAM-1 level in the bronchiolitis group (35.0+/-10.3%) was much higher than that in the bronchopneumonia (29.9+/-8.6%; p<0.05) and the control groups (24.6+/-6.9%; p<0.01). The bronchopneumonia group had higher ICAM-1 level than the control group (p<0.05). The RANTES level in the bronchiolitis (32.1+/-6.0 ng/mL) and the bronchopneumonia groups (30.6+/-6.2 ng/mL) was significantly higher than that in the control group (27.1+/-5.1 ng/mL) (p<0.01, p<0.05, respectively), however, no significant difference was found between the bronchopneumonia and bronchiolitis groups. There was a positive correlation between ICAM-1 and RANTES levels in the bronchiolitis group (r=0.675, P<0.01).

CONCLUSIONSICAM-1 and RANTES are involved in the pathogenesis of bronchiolitis and show a synergistic effect.

Bronchiolitis ; etiology ; metabolism ; Chemokine CCL5 ; analysis ; physiology ; Child, Preschool ; Female ; Humans ; Infant ; Intercellular Adhesion Molecule-1 ; analysis ; physiology ; Male