Up-regulation of p21 activated kinase 4 expression in the retina of diabetes mice and its effects on the behavior and mitochondrial function in retinal vascular endothelial cells / 中华眼底病杂志

Objective:To observe the effects of p21 activated kinase 4 (PAK4) on the mitochondrial function and biological behavior in retinal vascular endothelial cells.Methods:The experimental study was divided into two parts: in vivo animal experiment and in vitro cell experiment. In vivo animal experiments: 12 healthy C57BL/6J male mice were randomly divided into normal control group and diabetes group, with 6 mice in each group. Diabetes mice were induced by streptozotocin to establish diabetes model. Eight weeks after modeling, quantitative real-time polymerase chain reaction and Western blots were performed to detect the expression of PAK4 in diabetic retinas. In vitro cell experiments: the human retinal microvascular endothelial cells (hRMEC) were divided into three groups: conventional cultured cells group (N group), empty vector transfected (Vector group); pcDNA-PAK4 eukaryotic expression plasmid transfected group (PAK4 group). WB and qPCR were used to detect transfection efficiency, while scratching assay, cell scratch test was used to detect cell migration in hRMEC of each group. In vitro white blood cell adhesion experiment combined with 4 ', 6-diamino-2-phenylindole staining was used to detect the number of white blood cells adhering to hRMEC in each group. The Seahorse XFe96 cell energy metabolism analyzer measures intracellular mitochondrial basal respiration, adenosine triphosphate (ATP) production, maximum respiration, and reserve respiration capacity. The t-test was used for comparison between the two groups. Single factor analysis of variance was used for comparison among the three groups. Results:In vivo animal experiments: compared with normal control group, the relative expression levels of PAK4 mRNA and protein in retina of diabetic mice were significantly increased, with statistical significance ( t=25.372, 22.419, 25.372; P<0.05). In vitro cell experiment: compared with the N group and Vector group, the PAK4 protein, mRNA relative expression and cell mobility in the hRMEC of PAK4 group were significantly increased, with statistical significance ( F=36.821, 38.692, 29.421; P<0.05). Flow cytometry showed that the adhesion number of leukocytes on hRMEC in PAK4 group was significantly increased, and the difference was statistically significant ( F=39.649, P<0.01). Mitochondrial pressure measurement results showed that the capacity of mitochondrial basic respiration, ATP production, maximum respiration and reserve respiration in hRMEC in PAK4 group was significantly decreased, with statistical significance ( F=27.472, 22.315, 31.147, 27.472; P<0.05). Conclusion:Over-expression of PAK4 impairs mitochondrial function and significantly promotes leukocyte adhesion and migration in retinal vascular endothelial cells.

Role of lactadherin anti-idiotypic monoclonal antibodies Ab2β in the treatment of human rotavirus-induced diarrhea in a neonatal mouse model / 中国热带医学

@#Abstract: Objective To investigate the preventive and therapeutic effects of anti-idiotypic monoclonal antibodies (Ab2β) of lactadherin on neonatal mice infected with human rotavirus (HRV), and to analyze the underlying mechanism. Methods Hybridoma technology was used to prepare Ab2β of lactadherin. One hundred and twenty 7-day-old Kunming mice were randomly divided into groups A, B, C and control, each consisting of 30 mice. Groups A, B, and C were all infected with HRV via oral gavage. Group A received no treatment, group B was orally administered lactadherin for 7 days prior to infection, and group C was orally administered lactadherin for 7 days after infection, the control group was orally administered cell culture medium that did not contain the virus. The clinical manifestations (diarrhea, body weight) at different time points after infection of the neonatal mice in each group were observed, and the content of rotavirus antigen in the feces of neonatal mice was detected by enzyme-linked immunosorbent assay (ELISA). After HRV infection for 7 days, immunohistochemical staining was used to examine the expression level of intercellular adhesion molecule-1 (ICAM-1) in mouse small intestinal tissues in each group. Results No diarrhea occurred in the control group at any time point. Groups A, B, and C showed diarrhea symptoms after HRV challenge for 1 day. The degree of diarrhea in groups B and C was lower than that in group A at 2-4 days after HRV challenge, and the difference was statistically significant (P<0.05). The HRV antigen content in the feces of the neonatal mice in groups B and C was lower than that in group A at 1-5 days after HRV challenge, and the difference was statistically significant (P<0.05). There was no significant difference in the degree of diarrhea and HRV antigen content between groups B and C at each time point (P>0.05). There was no significant difference in the body weight of the neonatal mice in each group before infection and 1 day after infection (P>0.05); the weight of neonatal mice in groups B and C was higher than that in group A at 3, 5 and 7 days after HRV challenge, and the difference was statistically significant (P<0.05), and there was no significant difference in body weight between groups B and C at each time point after HRV challenge (P>0.05). The number of ICAM-1 expressing cells in the small intestine of the three groups A, B, and C was higher than that of the control group after HRV challenge for 7 days, and the difference was statistically significant (P<0.05). The cell number and gray value of ICAM-1 expressing cells in groups B and C were lower than those in group A, and the difference was statistically significant (P<0.05). Conclusions Anti-idiotypic monoclonal antibodies (Ab2β) of lactadherin has a good preventive and therapeutic effects on human rotavirus infection in neonatal mice, and can significantly improve diarrhea symptoms and reduce HRV viral load. Its specific mechanism may be related to the inhibition of ICAM-1.

Role of lactadherin anti-idiotypic monoclonal antibodies Ab2β in the treatment of human rotavirus-induced diarrhea in a neonatal mouse model / 中国热带医学

@#Abstract: Objective To investigate the preventive and therapeutic effects of anti-idiotypic monoclonal antibodies (Ab2β) of lactadherin on neonatal mice infected with human rotavirus (HRV), and to analyze the underlying mechanism. Methods Hybridoma technology was used to prepare Ab2β of lactadherin. One hundred and twenty 7-day-old Kunming mice were randomly divided into groups A, B, C and control, each consisting of 30 mice. Groups A, B, and C were all infected with HRV via oral gavage. Group A received no treatment, group B was orally administered lactadherin for 7 days prior to infection, and group C was orally administered lactadherin for 7 days after infection, the control group was orally administered cell culture medium that did not contain the virus. The clinical manifestations (diarrhea, body weight) at different time points after infection of the neonatal mice in each group were observed, and the content of rotavirus antigen in the feces of neonatal mice was detected by enzyme-linked immunosorbent assay (ELISA). After HRV infection for 7 days, immunohistochemical staining was used to examine the expression level of intercellular adhesion molecule-1 (ICAM-1) in mouse small intestinal tissues in each group. Results No diarrhea occurred in the control group at any time point. Groups A, B, and C showed diarrhea symptoms after HRV challenge for 1 day. The degree of diarrhea in groups B and C was lower than that in group A at 2-4 days after HRV challenge, and the difference was statistically significant (P<0.05). The HRV antigen content in the feces of the neonatal mice in groups B and C was lower than that in group A at 1-5 days after HRV challenge, and the difference was statistically significant (P<0.05). There was no significant difference in the degree of diarrhea and HRV antigen content between groups B and C at each time point (P>0.05). There was no significant difference in the body weight of the neonatal mice in each group before infection and 1 day after infection (P>0.05); the weight of neonatal mice in groups B and C was higher than that in group A at 3, 5 and 7 days after HRV challenge, and the difference was statistically significant (P<0.05), and there was no significant difference in body weight between groups B and C at each time point after HRV challenge (P>0.05). The number of ICAM-1 expressing cells in the small intestine of the three groups A, B, and C was higher than that of the control group after HRV challenge for 7 days, and the difference was statistically significant (P<0.05). The cell number and gray value of ICAM-1 expressing cells in groups B and C were lower than those in group A, and the difference was statistically significant (P<0.05). Conclusions Anti-idiotypic monoclonal antibodies (Ab2β) of lactadherin has a good preventive and therapeutic effects on human rotavirus infection in neonatal mice, and can significantly improve diarrhea symptoms and reduce HRV viral load. Its specific mechanism may be related to the inhibition of ICAM-1.

Interferon gene stimulating protein inhibitor improves leukocyte adhesion and glycolysis of retinal vascular endothelial cells / 中华眼底病杂志

Objective:To investigate the effects of interferon gene stimulating protein (STING) inhibitor (C176) on human retinal microvascular endothelial cells (hRMEC) under oxidative stress.Methods:An animal experimental study. In vivo experiment: 48 healthy male C57BL/6J mice were randomly divided into wild type mice group (WT group) and diabetes (DM) group, with 24 mice in each group. DM mice were induced by streptozotocin to establish DM model. After successful modeling, DM group was divided into DM+dimethyl sulfoxide (DMSO) group and DM+C176 group, with 12 mice in each group. The mice in the DM+DMSO group were intraperitoneally injected with DMSO at the dose of 50 mg/kg. Mice in DM+C176 group were intraperitoneally injected with STING inhibitor C176 750 nmol at the dose of 50 mg/kg. Four weeks after modeling, immunohistochemical staining, Western blot and real-time fluorescence quantitative polymerase chain reaction were used to detect the expression of STING in the retina of WT and DM mice. The leukocyte adhesion test was used to detect the number of leukocytes adhering to hRMEC in mice with WT, DM+DMSO and DM+C176 groups. In vitro experiment: hRMEC was randomly divided into conventional culture cell group (N group), dimethyl sulfoxide (DMSO) group (with DMSO intervention) and C176 group (with C176 intervention). The cells were induced by 150 μg/ml glycation end products for each group. In vitro leukocyte adhesion test combined with 4', 6-diamino-2-phenylindole staining was used to detect the number of leukocytes adhering to hRMEC. The adherent leukocytes were quantitatively analyzed by flow cytometry; H 2DCFDA/reactive oxygen species (ROS) fluorescence probe was used to detect ROS expression in cells; Seahorse XFe96 cell energy metabolism analyzer was used to measure the level of intracellular glycolysis. t-test was used to compare the two groups; single factor analysis of variance was used to compare the three groups. Results:In vivo experiment: compared with WT group, the expression level of STING ( t=73.248) and the relative expression amount of mRNA ( t=67.385) in the retina of DM group mice increased significantly ( P<0.05). Compared with WT group, the number of leukocytes adhering to the retinal vessels of mice in DM+DMSO group was significantly increased, while that in DM+C176 group was significantly decreased ( F=84.352, P<0.01). In vitro: compared with N group and DMSO group, the number of leukocyte adhesion on hRMEC in C176 group decreased significantly ( F=35.251, P<0.01). Compared with N group, the number of leukocytes adhering to hRMEC in DMSO group and C176 group decreased significantly ( F=26.374, P<0.01). The ROS level in hRMEC in C176 group was significantly lower than that in N group and C176 group ( F=41.362, P<0.01). Compared with N group and DMSO group, the glycolysis level of hRMEC in C176 group was significantly reduced, with a statistically significant difference ( F=68.741, P<0.01). Conclusion:Inhibiting the expression of STING in retinal vascular endothelial cells can improve the progress of DM by inhibiting leukocyte adhesion, ROS production and glycolysis level.

Carrier frequency of autosomal recessive disorders (BC, BLAD, FXID and CVM) in Holstein cows in Jalisco, Mexico

The hereditary autosomal recessive disorders bovine citrullinemia (BC), bovine leukocyte adhesion deficiency (BLAD), factor XI deficiency (FXID), and complex vertebral malformation (CVM) have affected dairy cattle breeding significantly around the world. This study examined the carrier frequency of BC, BLAD, FXID, and CVM autosomal recessive disorders in Bos taurus Holstein cows bred in the Altos Norte region of the state of Jalisco, Mexico. We extracted DNA from 408 random samples of peripheral blood, and then used polymerase chain reaction (PCR) to identify insertion mutations for FXID, and PCR with restriction fragment length polymorphism (PCR-RFLP) for CVM, BC and BLAD. We visualized the PCR products using agarose gel electrophoresis stained with GelRed®. We found that 100% of wild-type (N/N) allele homozygous animals for genes CD18, ASS, and FXI were free of the mutations for BLAD, BC and FXID respectively. For gene SLC35A3 we estimated total carrier frequency of 10.3% and allele frequency of 5%.(AU)

Leukocyte Adhesion Deficiency Associated with Neonatal Septic Hip in a Late Preterm Infant

Leukocyte adhesion deficiency is a rare primary immunodeficiency and autosomal recessive disorder caused by a mutation in the gene encoding CD18, which is a constituent of leukocyte integrins. Clinical features usually begin with a delay in the separation of the umbilical cord in the neonatal period, and are characterized by marked leukocytosis with infection, delayed wound healing, and repeated bacterial and fungal infections. We experienced a case of leukocyte adhesion deficiency diagnosed in the neonatal period, in which a late preterm infant admitted to neonatal intensive care unit presented with a septic hip. Flow cytometry analysis of whole blood showed a decrease in the expression of CD11b/CD18. This is the first case of leukocyte adhesion deficiency with neonatal septic hip diagnosed in Korea.

Congenital defects of phagocyte number or function / 中华实用儿科临床杂志

Congenital defects of phagocyte number,function or both was categorized to the fifth classification from the international union of immunological societies expert committee for primary immunodeficiency 2015.Severe congenital neutropenia was the most fatal phagocyte number defect.Phagocyte functions included motility,chemotaxis,adhesion,phagocytosis and killing.Leukocyte adhesion deficiency and chronic granulomatous disease were the most common diseases.This article will describe pathogenesis,molecular,clinical,laboratory features and treatment and prognosis,to supporting clues for paediatrician's clinical operations.

Adesão leucocitária e MDA sérico em cães naturalmente infectados por Leishmania infantum / Leukocyte adhesion and MDA serum levels in naturally infected dogs by Leishmania infantum

O presente estudo avaliou o estresse oxidativo e a adesão de leucócitos (AL) em cães naturalmente infectados por Leishmania infantum. Foram utilizados cães saudáveis (CN = 10) e cães acometidos por leishmaniose visceral na forma sintomática (CS = 10), submetidos previamente a exames de imunofluorescência indireta (IFI), ensaio imunoenzimático (ELISA) e pesquisa do parasito em aspirados de medula óssea. Soro foi utilizado para avaliação de malondialdeído (MDA) no ensaio para espécies reativas ao ácido tiobarbitúrico (TBARS) e AL foi determinada pelo método da coluna de náilon em sangue em EDTA, heparina e citrato. Os dados de AL foram expressos em porcentagem e MDA em média ± desvio padrão, submetidos ao teste T de student não pareado (p < 0,05). Amostras em heparina apresentaram níveis mais elevados de AL no grupo CS (55,62%, p < 0,05) quando comparadas com EDTA e citrato (10,46% e 5,28%). Citrato e EDTA inibiram AL em cães doentes e saudáveis, enquanto a heparina preservou a AL. A proporção neutrofílica se apresentou reduzida nas amostras em heparina (85% para 67%, p <0,05) quando comparadas com citrato e EDTA, que por sua vez mantiveram-se estáveis (83% para 80%). Os níveis de MDA apresentaram-se mais elevados em CS (0,0117µM ± 0,002) quando comparado com CN (0,0057µM ± 0,001) (p <0,05). Estes dados dão suporte à conclusão de que na LVC ocorre elevação do estresse oxidativo e aumento da expressão das moléculas de adesão nos leucócitos, evidenciando resposta inflamatória sistêmica. A escolha do anticoagulante é importante para a implementação do ensaio de AL.

The present study evaluated the oxidative stress and leukocyte adhesion (LA) in dogs naturally infected by Leishmania infantum. Healthy dogs (HD = 10) and dogs affected by canine visceral leishmaniasis (CVL) in symptomatic form (SD= 10) were previously submitted to indirect immunofluorescence reaction (IIF), enzyme-linked immunosorbent assay (ELISA) and detection of parasites in bone marrow aspirates. Serum was used to assess malondialdeid (MDA) by thiobarbituric acid reactive species (TBARS) assay and AL was determined by nylon column method using whole blood stored in EDTA, citrate and heparin. AL data was expressed as percentage and MDA data as mean ± standard deviation, both submitted to the unpaired student's T test (p < 0.05). Heparin samples showed higher levels of AL in CS group (55.62 %, p < 0.05) when compared with EDTA and citrate (10.46% and 5.28%). Citrate and EDTA inhibit AL in healthy and sick dogs, while heparin preserved AL in both groups. Neutrophil proportion in heparin samples were lower (85% to 67%, p <0.05) compared with citrate and EDTA, which in turn remained practically unchanged (83% to 80%). MDA levels were higher in SD (0.0117µM ± 0.002) when compared to HD (0.0057µM ± 0.001) (p <0.05). These data support the conclusion that CVL induces oxidative stress enhance and leukocyte adhesion increase, indicating systemic inflammatory response. The choice of anticoagulant is an important decision for implementing LA assays.

Suppression effect of MaiShu on formation of atherosclerotic plaque of apolipoprotein E knock-out mice / 药学学报

The research aimed to investigate the suppression effect of MaiShu which contains hawthorn, hippophae, medlar, phytosterols (β-sitosterol, stigmasterol and campesterol), β-glucan and lycopeneon formation of atherosclerotic plaque in apolipoprotein E knock-out (ApoE-/-) mice. Liquid chromatography-ultraviolet-mass spectrometry (LC-UV-MC) methods were used to analyze the main chemical composition of MaiShu.Atherosclerotic mice models were established by high-fat diet. The mice were administrated with MaiShu (1, 2, 4 g·kg-1·d-1) or other contrast materials by intragastric route for 10 weeks continuously. At the end of administration, the blood of mice was collected for tests of the serum total cholesterol (TC), total triglyceride (TG) and high density lipoprotein cholesterol (HDL-C) level. Atherosclerotic lesions in aorta and aortic root were assessed by calculating the relative area of lesions (oil red O stained). Intravital fluorescence microscopic system was used to evaluate the leukocyte-endothelial adhesion in mesenteric artery of mice by detecting the rolling velocity of white blood cells (WBC). Collagenous fibers and macrophages in lesions were detected by sirius red staining and immunological histological chemistry to evaluate the atherosclerotic plaque stability. Results showed that MaiShu contains various flavonoids (9.5%), phytosterols (23.8%) and polysaccharides (8.9%). The serum lipid level of model animals was significantly higher than the control animals. Serum TC level was decreased by MaiShu (4 g·kg-1, P-1) compared to model (P-1, P-1, P-1, P-1, P-1, P-1) and macrophages were decreased (2, 4 g·kg-1) compared to model. These results demonstrate that MaiShu can obviously decrease the serum lipid levels and the risk of leukocyte-endothelial adhesion in ApoE-/- mice. The effect of MaiShu may be associated with the decrease of macrophages in plaque.

Quality evaluation and specification research of transfer factor injections / 中国药学杂志

OBJECTIVE: To understand the quality status and problems of domestic transfer factor injections and evaluate the suitability of statutory standard and additional explorative researches. METHODS: Fifty-seven batches of transfer factor injections were analyzed according to the legal quality standard and the results were statistically analyzed to assess the overall level of quality. The qualities of the products form different manufacturing enterprises were compared. According to the explorative researches, the overall situation of product quality and standard condition were then comprehensively evaluated. RESULTS: The qualified rate of the 57 batches of samples was 91.2%. Four samples failed due to visible foreign substances and one sample failed due to unqualified content of ribose. There were obvious differences between the results of legal test and exploratory research. The results reflected some defects and problems of the legal quality standard which was greatly improved and revised in our research. CONCLUSION: The overall quality status of transfer factor injections is not ideal. The results of exploratory research demonstrate that many defects exist in the legal quality standard and a series of exclusive, accurate and sensitive methods should be established to fully control the quality of transfer factor injections.

Clinical Profile of Leukocyte Adhesion Deficiency Type I.

Leukocyte adhesion deficiency type I (LAD-I) is a rare, inherited immunodeficiency with defect in the recruitment of leukocyte to the site of inflammation. Patients with severe LAD-I have absent or markedly reduced expression of CD18 and CD11. Here we report clinical profile of 7 cases of LAD-I diagnosed at our center over a period of 3 years. Recurrent skin and mucous membrane infections were the major presenting manifestations. All children had a history of delayed cord separation.

Leukocyte adhesion deficiency syndrome: report on the first case in Chile and South America / Síndrome de deficiencia de adhesión leucocitaria: informe del primer caso en Chile y América del Sur

CONTEXT: Adhesion molecule deficiency type 1 is a rare disease that should be suspected in any patient whose umbilical cord presents delay in falling off, and who presents recurrent severe infections. Early diagnostic suspicion and early treatment improve the prognosis. CASE REPORT: The case of a four-month-old boy with recurrent hospitalizations because of severe bronchopneumonia and several episodes of acute otitis media with non-purulent drainage of mucus and positive bacterial cultures is presented. His medical history included neonatal sepsis and delayed umbilical cord detachment. Laboratory studies showed marked leukocytosis with predominance of neutrophils and decreased CD11b and CD18. These were all compatible with a diagnosis of leukocyte adhesion deficiency type I [LAD type 1].

CONTEXTO: El deficit de moleculas de adhesión tipo 1 es una enfermedad rara que debe ser sospechada en todo paciente que presente un retardo en la caída del cordón unmbilical, además de infecciones graves a repetición. Un sospecha diagnóstica precoz y un tratamiento oportuno mejorarán el pronóstico. INFORMES DE CASO: Se presenta el caso de un niño de cuatro meses de edad, con hospitalizaciones recurrentes a causa de bronconeumonía severa y varios episodios de otitis media aguda con drenaje mucoso, no purulento, y cultivos bacterianos positivos. Su historial médico incluye la sepsis neonatal y el desprendimiento tardío del cordón umbilical. Los estudios de laboratorio mostraron leucocitosis marcada con predominio neutrofílico y disminución de CD11b y CD18, todos compatibles con el diagnóstico del tipo de deficiencia de adhesión leucocitaria 1 [tipo LAD 1].

A novel femur window chamber for in vivo studies of bone microcirculation

Background: Due to technical difficulties of in vivo observation of blood flow and microvessels in bone, no study has been done concerning the role of blood flow in bone remodeling. Objective: To develop a new window chamber for microscopic observation of the microcirculation in living bone, and to examine the utility of the chamber using rat femur in health and diseases. Methods: A stainless chamber (19 mm in diameter and 5 mm in height) with a circular window (7.5 mm in diameter) for microscopic observation was developed. The chamber was put on rat femur which was exposed for direct observation of the microvasculature. Intravital observation was made of bone blood flow and microvessels, using fluorescence videomicroscopy and confocal laser microscopy. The utility of the chamber was examined based on images of microcirculation (normal and abnormal) in the femur bone. Results and conclusions: Images of rat femur microvasculature were enhanced in the quality by use of the femur window chamber. The new chamber provides a powerful tool for in vivo studies of the bone microcirculation in health and diseases.

Research progress of carbon monoxide protection mechanism / 中国病理生理杂志

Carbon monoxide(CO) has been considered as a fatal gas.Recent researches show that tissues generate carbon monoxide,and carbon monoxide plays a key role in the tissue physiological and pathological processes.The endogenous CO is generated from the decomposition of heme which is catalyzed by heme oxygenase enzyme(HO).Studies show that CO involves in the process of anti-inflammation,anti-apoptosis,regulation of vascular tone,modulation of leukocyte adhesion and platelet aggregation.CO also functions as neurotransmitters.

Platelet-activating Factor Mediates Hydrogen Peroxide Induced Endothelial-leukocyte Adhesion / 第二军医大学学报

The effects of hydrogen peroxide (H2O2) on endothelial-polymorphonuclear cells (EC-PMN) adhesion and their mechanisms wsre studied in cultured bovine pulmonary artery endothelial monolayers in vitro. H2O2 at various concentrations (10-1, 10-2, 10-3mol/L respectively) stimulated EC dependent PMN adhesion, of which l02mol/L H2O2 was the most potent one, increasing adhesion to 2.3 times that of the control. Pretreatment of PMNs with SRI 63-441, a platelet-activating factor (PAF) receptor antagonist, had no effect on H2O2 induced EC-PMN adhesion. Pretreatment of ECs with SRI 63-441 before H2O2 exposure significantly decreased PMN adherence to ECs. Pretreatment of ECs with phospholipase A2 inhibitor p-bromophenacyl-bromide or cahnodulin antagonist chlorpromazine and aildum ion chelate EGTA obviously decreased H2O2 induced increment of EC-PMN adhesion. These results suggest that H2O2 may activate ECs, causing the inflow of extracellular calcium or the release of calcium from intracellular deposits. Increased intracellular Ca2+ may bind with calmodulin to activate phospholipase A2 thus initiating PAF synthesis and promoting EC-PMN adhesion.

Inhibitory Effects of Protein Kinase C Inhibitors on Polymorphoneuclear-endothelial Cell Adhesion / 第二军医大学学报

The effects of protein kinase C (PKC) inhibitors 1- (5-isoquinolinylsulfonyl) - 2-methylpoperazine (H-7) and quercetin on endotheliai-polymorphoneuclear (EC-PMN) adhesion induced by tumor necrosis factor (TNF) and platelet activating factor (PAF) were studied in cultured bovine pulmonary artery endothelial monolayers in vitro. TNF (100 U/ml) and PAF (1.0 ?mol/L) stimulated EC dependent PMN-EC adhesion. Both H-7 and quercetin dose-dependently inhibited TNF and PAF induced PMN-EC adhesion. The IC50 of H-7 was 22.22, 5.25 umol/L, and that of quercetin was 18.30, 4.83 ?mol/L respectively. WEB-2086, a specific PAF receptor antagonist, dose-dependently inhibited PAF induced PMN-EC adhesion, but had no effect on TNF induced adhesion. These results suggest that PKC play an important role in EC activation by TNF or PAF, and TNF induced PMN-EC adhesion by independent on endogenetic PAF.