Animals ; Endothelium, Vascular ; metabolism ; Eukaryotic Cells ; metabolism ; Genetic Therapy ; Neoplasms ; blood supply ; therapy ; Neovascularization, Pathologic ; Plasmids ; Receptors, Cell Surface ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Swine

OBJECTIVETo evaluate variations in the plasma tissue factor (TF) and urokinase type plasminogen activator (u-PA) system and their relationship with clinical cancer type, pathological classification and metastatic status in cancer patients.

METHODSPlasma levels of TF and its inhibitor (TFPI), as well as u-PA and its receptor (u-PAR) were measured using ELISA in 76 patients with malignant tumors and 24 patients with benign tumors.

RESULTSPlasma levels of TF and u-PAR in the malignant tumor group were significantly higher than those of the benign tumor group and the normal control. U-PA and u-PAR increased significantly in patients with esophageal and gastric cancer. However, most of these parameters except TFPI did not vary according to pathological classification. A significant elevation was evident in patients with local infiltration, lymph node involvement and distal metastasis, while u-PAR only increased in the latter two categories.

CONCLUSIONSBoth the TF and u-PA systems are activated in cancer patients. U-PA and its receptor might prove to be a clinically useful marker for disease progression.

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Lipoproteins ; blood ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplasms ; blood ; pathology ; Receptors, Cell Surface ; blood ; Receptors, Urokinase Plasminogen Activator ; Thromboplastin ; metabolism ; Urokinase-Type Plasminogen Activator ; blood

Interleukin (IL)-33 is an important mediator of innate immunity. Behcet's disease (BD) is an autoinflammatory disorder characterized by hyperactivity of the innate immune response. We measured serum levels of IL-33 and its receptor soluble ST2 (sST2) in patients with BD to investigate their association with disease activity. Serum levels of both IL-33 and sST2 were higher in patients with BD compared with those in normal controls (IL-33: 594.48+/-175.04 pg/mL in BD and 224.23+/-56.64 pg/mL in normal controls [P=0.048], sST2: 99.01+/-15.92 pg/mL in BD and 23.56+/-3.25 pg/mL in normal controls [P<0.001]). IL-33 and sST2 expression in skin tissue, as shown by immunohistochemistry, was higher in patients with BD compared with that in the normal controls. Serum sST2 level correlated significantly with the BD currently active form (BDCAF), Iranian BD dynamic activity measure (IBDDAM), erythrocyte sedimentation rate and C-reactive protein. Multiple linear regression showed that serum sST2 was an independent factor associated with IBBDAM (regression coefficient, 0.374; P=0.004), and BDCAF (regression coefficient, 0.236; P=0.047). These results demonstrate that IL-33 and sST2 are highly expressed in patients with BD and that serum sST2 is an independent factor associated with IBDDAM and BDCAF, suggesting a potential role for sST2 as a surrogate marker of disease activity in patients with BD.
Adult ; Behcet Syndrome/blood/*pathology ; Blood Sedimentation ; C-Reactive Protein/analysis ; Female ; Humans ; Immunohistochemistry ; Interleukins/*blood/metabolism ; Male ; Middle Aged ; Receptors, Cell Surface/*blood/metabolism ; Severity of Illness Index ; Skin/metabolism/pathology

OBJECTIVESTo explore the role of TLR4 in the mechanism of hepatic ischemia/reperfusion (I/R) injury in mice.

METHODSWild-type (C3H/Heouj) mice and TLR4 deficient mice (C3H/Hej) were used to prepare the models of liver I/R injury. Partial hepatic ischemia was produced by inflow causing occlusion in the median and left lobes for 45 minutes. Blood was drawn to kill the mice at 1 hours and 3 hours after reperfusion. The blood was used to analyze aspartate aminotransferase (AST) and tumor necrosis factor-alpha (TNFalpha). TNF-alpha mRNA expression and myeloperoxidase (MPO) level in ischemic lobes was examined by northern blot and myeloperoxidase assay, respectively.

RESULTSAST levels were significantly lower in TLR4 deficient mice, compared with those in wild-type mice at both time points (661.83U/L+/-106.09U/L vs. 1215.5U/L+/- 174.03U/L, t=-6.65, P<0.01; 1145.17U/L+/-132.42U/L vs. 2958.17U/L+/-186.81U/L, t=-5.57, P<0.01). Serum TNF-alpha level was lower in TLR4 deficient mice at 3 hours after reperfusion compared with that in wild-type mice (152.39pg/ml+/-43.3 pg/ml vs. 249.12pg/ml+/-51.89pg/ml, t=-3.13, P<0.05). This difference appeared to be mediated at the gene level, since TNF-alpha mRNA expression had decreased in TLR4 deficient mice at 1 hours after reperfusion, compared with that in wild type mice (80.3+/-28.8 vs. 189.4+/-24.6, t=-3.25, P<0.05). MPO level in ischemic lobes in TLR4 deficient mice at 3 hours after reperfusion was significantly lower than that in wild type mice (F=33.49, P<0.01).

CONCLUSIONSI/R hepatic injury in TLR4 deficient mice is less than that in wild-type mice. TNF-alpha expression down-regulated at the mRNA level appears critical. These suggest that TLR4 be involved in the mechanism of hepatic ischemia/reperfusion injury in mice.

Alanine Transaminase ; blood ; Animals ; Liver ; blood supply ; metabolism ; Membrane Glycoproteins ; physiology ; Mice ; Mice, Inbred C3H ; Peroxidase ; metabolism ; RNA, Messenger ; analysis ; Receptors, Cell Surface ; physiology ; Reperfusion Injury ; etiology ; Toll-Like Receptor 4 ; Toll-Like Receptors ; Tumor Necrosis Factor-alpha ; analysis ; genetics

This project was aimed to investigate the role and the underlying mechanism of microglia polarization on blood-brain barrier (BBB) during cerebral ischemia-reperfusion. After construction of the mouse model of cerebral ischemia-reperfusion, upregulated IL-6 and TNF-α in peripheral blood and increased IL-6 and iNOS in ischemia tissues were confirmed. The supernatant expression of TNF-α and IL-6, as well as IL-6, iNOS and CD86 mRNA, was significantly increased in the of Bv-2 cells after oxygen-glucose deprivation/reoxygenation (OGD/R) model was constructed in vitro. For further understanding the expression pattern of RNAs, the next-generation RNA sequencing was performed and upregulation of Robo4 (roundabout guidance receptor 4) was found both in M1-polarized and OGD/R treated Bv-2 cells, which was also confirmed by RT-qPCR. Extracellular soluble Robo4 (sRobo4) protein also increased in the supernatant of M1-polarized and OGD/R treated Bv-2 cells. Treating bEND3 cells with the Robo4 recombinant protein, M1-polarized Bv-2 cell supernatant or OGD/R Bv-2 cell supernatant decreased trans-endothelial electrical resistance (TEER), suggesting the injury of BBB. In addition, Robo4 was also highly expressed in the serum of patients who experienced acute ischemia stroke and mechanical thrombectomy operation. All the results suggest that increased secretion of Robo4 by M1-polarized-microglia during cerebral ischemia-reperfusion is most likely one of the causes of BBB injury, and Robo4 may be one of the therapeutic targets for BBB functional protection.
Animals ; Blood-Brain Barrier/metabolism* ; Brain Ischemia/drug therapy* ; Glucose/metabolism* ; Interleukin-6/metabolism* ; Mice ; Microglia/metabolism* ; Oxygen/metabolism* ; Receptors, Cell Surface/metabolism* ; Reperfusion ; Reperfusion Injury/drug therapy* ; Tumor Necrosis Factor-alpha/metabolism*

Hypercoagulable state and thrombosis are major lethal causes of ulcerate colitis (UC). The aim of the present study is to explore the change and role of protein C (PC) system in UC thrombosis. 4% dextran sulfate sodium (DSS) was used to induce the UC model, and the body weight, the length of colon, and the weight of spleen were measured after intake of DSS as drinking water for 1 week. The macroscore and microscore were examined. The quantity of macrophage in colon smooth muscle was observed by immunofluorescence, and TNF-α and IL-6 levels in plasma were evaluated by ELISA. Intravital microscopy was applied to observe colonic mucosal microvascular circulation, activities of PC and protein S (PS) were determined by immunoturbidimetry, endothelial cell protein C receptor (EPCR) and thrombomodulin (TM) expressions were detected by immunohistochemistry. In vitro, TNF-α and IL-6 levels were tested in supernatant of macrophage separated from colonic tissue. After stimulation of mouse colonic mucosa microvascular endothelial cells by TNF-α and IL-6 respectively, the activities of PC, PS, activated protein C (APC) were evaluated, and the expressions of EPCR and TM were detected by Western blotting. The results revealed that compared with control, the DSS mouse showed weight loss (P < 0.05), a shortened colon (P < 0.05), and swelled spleen (P < 0.05), accompanied by higher histological score (P < 0.05), as well as infiltration of macrophages, elevated TNF-α and IL-6 levels in plasma (P < 0.01). The intravital microscopy results revealed that compared with control, DSS mice showed significantly enhanced adhesion of leukocytes and colonic mucosal microvascular endothelial cells (P < 0.01), meanwhile, decreased activity of PC and PS in plasma (P < 0.01 or P < 0.05), and down-regulated expression of EPCR (P < 0.01). The degree of inflammation was negatively correlated with the PC activity. In vitro, TNF-α and IL-6 levels were increased in the supernatant of macrophages from DSS mice colonic tissue (P < 0.05), and after incubation of TNF-α or IL-6 with colonic mucosal microvascular endothelial cells, the APC activity was decreased (P < 0.05 or P < 0.01), and expression of EPCR was down regulated (P < 0.05). These results suggest that PC system is inhibited in UC mouse. Presumably, the mechanism may be due to the secretion of cytokines from macrophages and subsequential influence on the function of endothelia cells. Furthermore, enhancement of PC system activity may serve as a new strategy for the treatment of UC.
Animals ; Blood Coagulation Factors ; metabolism ; Colitis, Ulcerative ; chemically induced ; physiopathology ; Dextran Sulfate ; Immunohistochemistry ; Inflammation ; Interleukin-6 ; blood ; Intestinal Mucosa ; pathology ; Macrophages ; cytology ; Mice ; Protein C ; metabolism ; Receptors, Cell Surface ; metabolism ; Spleen ; pathology ; Tumor Necrosis Factor-alpha ; blood

Ligand-grafted long circulation liposome is an advanced and effective carrier in which drugs are modified to be stabilized in the body and delivered properly in time and amount at targeted organs with suppressing side effects.
Animals ; Chemistry, Pharmaceutical ; methods ; Drug Carriers ; chemistry ; Drug Delivery Systems ; Drug Stability ; Humans ; Ligands ; Liposomes ; blood ; chemistry ; Nuclear Medicine ; methods ; Phosphatidylethanolamines ; chemistry ; Polyethylene Glycols ; chemistry ; Receptors, Cell Surface ; metabolism ; Surface Properties

OBJECTIVETo investigate the possibility of microvessel density (MVD) and blood vessel invade (BVI) as the indexes in predicting prognosis of rectal carcinoma at stages I to II.

METHODSTumor tissues from 380 patients who underwent resection of stage I or II rectal cancer were analyzed for MVD and BVI by immunohistochemical S-P method with anti-CD105 and anti-CD 34 antibody. Binary and multivariable Cox regression was applied to indicate independent factors associated with overall survival.

RESULTSCD105 was present in the neovascularity of the cancer tissue but not in the normal tissue, while CD34 was present in the tumor tissue and the normal tissue. BVI on CD34 staining was significantly higher than that on HE staining. Multivariable analysis revealed that TNM stage, CD34-BVI, histologic type, and CD105-MDV were independent risk factors to predict the possibility of poor prognosis of stage I or II rectal cancer. CD34-BVI or CD105-MVD positivity had a hazard ratio of 4.483 (95% confidence interval 2.861-7.026) for mortality.

CONCLUSIONThe expressions of CD34-BVI and CD105-MVD are independent factors to predict the possibility of poor survival of stage I or II rectal carcinoma. Detection of CD105-MVD combined with CD34-BVI may help predict clinical outcome and design further individualized adjuvant treatment.

Adult ; Aged ; Aged, 80 and over ; Antigens, CD ; metabolism ; Antigens, CD34 ; metabolism ; Endoglin ; Female ; Humans ; Male ; Microvessels ; pathology ; Middle Aged ; Neoplasm Staging ; Neovascularization, Pathologic ; pathology ; Prognosis ; Receptors, Cell Surface ; metabolism ; Rectal Neoplasms ; blood supply ; diagnosis ; pathology

OBJECTIVE: To explore the expressions of thymidine phosphorylase (TP), midkine (MK) and MVD marked with CD105 antibody in laryngeal squamous cell carcinoma (LSCC) and their clinical significance. METHOD: The expressions of TP, MK and CD105 in LSCC tissues of 43 cases were studied by immunohistochemical staining. RESULT: The positive expression rates of TP and MK in LSCC were 67.4% and 60.5% respectively, the mean value of MVD was 6.01 +/- 1.78. MVD was significantly higher in tumor tissue with both positive TP and MK than in that with both negative TP and MK (7.07 +/- 3.26 vs. 4.03 +/- 1.90, P < 0.05). The expression of TP, MK and CD105 were all correlated with T-stage and lymph node metastasis. Positive TP, MK expression and high MVD were all associated with a poor survival, and positive expression of both TP and MK in tumors conferred a poorer prognosis than negative expression of those factors in tumors, but only the lymph node metastasis and MVD were independent prognostic factors on multivariate analysis. CONCLUSION Both TP and MK are important for angiogenesis in LSCC. TP, MK and angiogenesis are all closely correlated with the progress of LSCC and the lymph node metastasis. The lymph node metastasis and MVD marked with CD105 antibody were independent prognostic factors. TP and MK may affect the progression and prognosis of tumor by promotion of angiogenesis. A combinative detection of TP, MK and CD105 can be as valuable tumor marker and prognostic factor for LSCC.
Adult ; Aged ; Antigens, CD ; metabolism ; Carcinoma, Squamous Cell ; blood supply ; metabolism ; pathology ; Endoglin ; Female ; Humans ; Laryngeal Neoplasms ; blood supply ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Midkine ; Neoplasm Staging ; Neovascularization, Pathologic ; Nerve Growth Factors ; metabolism ; Prognosis ; Receptors, Cell Surface ; metabolism ; Thymidine Phosphorylase ; metabolism

Adenocarcinoma ; blood supply ; metabolism ; mortality ; secondary ; Adult ; Aged ; Female ; Humans ; Lymphatic Metastasis ; Male ; Microcirculation ; Middle Aged ; Neovascularization, Pathologic ; metabolism ; pathology ; Prognosis ; RNA, Messenger ; biosynthesis ; genetics ; Receptors, Cell Surface ; biosynthesis ; genetics ; Receptors, Urokinase Plasminogen Activator ; Stomach Neoplasms ; blood supply ; metabolism ; mortality ; pathology ; Survival Rate ; Vascular Endothelial Growth Factor A ; metabolism