EFFECTS OF RED LIGHT AT 640nm FROM LIGHT EMITTING DIODES ON THE RESPIRATORY BURST OF HUMAN NEUTROPHILS

Photobiomodulation(PBM)has been reported to have effects on respiratory burst of polymorphonuclear neutrophils(PMNs),but little focus was on the individual differences of human PMNs.The latter was investigated in this study.The PMNs were isolated from peripheral blood of 13 volunteers(10 ordinary persons,3 athletes)and treated by red light(640±15 nm)from light emitting diodes(RLED)at 50,100,300,500 and 1000 J/m^(2)for 100 seconds,respectively.Blood samples of athletes were extracted at different running stages in 10 km non-interrupted long-distance running,before running,1 hour after running began,just finishing the running,resting for 1 hour and 2 hours after running.The PMN respiratory burst was assessed by the nitroblue tetrazolium test.It was found that there were three types of RLED PBM on the respiratory burst of 3 types of PMNs,respectively,inducing for the subactivated PMNs,inhibiting for the overactivated PMNs and none for the PMNs in homeostasis.It was then concluded that there may be RLED PBM on dysfunctional human PMNs while none on those in homeostasis,and RLED at 300 J/m^(2)for 100 seconds may have bi-direction modulation on PMN respiratory burst.

The Observation of Complement Activation and Polymorphonuclear Neutrocytopenia during Cardiopulmonary Bypass

By determining the plasma levels of C3, C4, factor B and polymorphonuclear neutrophils (PMNs) of the patients who received CPB, the path of complement activation and changes of PMNs were studied. The results suggest that complement system was activated through alternative pathway during CPB and was activated through classic pathway after CPB. The anaphylatoxin, the products of complement activation may be responsible for the polymorphonuclear neutrocytopenia.

Macrophage inflammatory protein-2 as mediator of inflammation in acute liver injury

Macrophage inflammatory protein(MIP)-2 is one of the CXC chemokines and is also known as chemokine CXC ligand(CXCL2). MIP-2 affects neutrophil recruitment and activation through the p38 mitogen-activatedprotein-kinase-dependent signaling pathway, by binding to its specific receptors, CXCR1 and CXCR2. MIP-2 is produced by a variety of cell types, such as macrophages, monocytes, epithelial cells, and hepatocytes, in response to infection or injury. In liver injury, activated Kupffer cells are known as the major source of MIP-2. MIP-2-recruited and activated neutrophils can accelerate liver inflammation by releasing various inflammatory mediators. Here, we give a brief introduction to the basic molecular and cellular sources of MIP-2, and focus on its physiological and pathological functions in acute liver injury induced by concanavalin A, lipopolysaccharides, irradiation, ischemia/reperfusion, alcohol, and hypoxia, and hepatectomy-induced liver regeneration and tumor colorectal metastasis. Further understanding of the regulatory mechanisms of MIP-2 secretion and activation may be helpful to develop MIP-2-targeted therapeutic strategies to prevent liver inflammation.

High-mobility group box1 as an amplifier of immune response and target for treatment in Aspergillus fumigatus keratitis

AIM:To determine the roles of high-mobility group box1(HMGB1)in pro-inflammation,host immune response and its potential target for treatment in Aspergillus fumigatus(A.fumigatus)keratitis.METHODS:Expression of HMGB1 was tested in C57 BL/6 normal and infected corneas.Dual immunostaining tested coexpression of HMGB1 with TLR4 or LOX-1.C57 BL/6 mice were pretreated with Box A or PBS and then infected.Clinical scores,polymerase chain reaction,ELISA,and MPO assay were used to assess the disease response.Flow cytometry were used to test the effect of Box A on reactive oxygen species(ROS)expression after A.fumigatus stimulation in polymorphonuclear neutrophilic leukocytes(PMN).C57 BL/6 peritoneal macrophages were pretreated with Box B before A.fumigatus stimulation,and MIP-2,IL-1β,TNF-α,HMGB1 and LOX-1 were measured.Macrophages were pretreated with Box B or Box B combined with Poly(I)(an inhibitor of LOX-1)before stimulating with A.fumigatus,and MIP-2,IL-1β,TNF-α,LOX-1,p38-MAPK,p-p38-MAPK were measured.RESULTS:HMGB1 levels were elevated in C57 BL/6 mice after infection.HMGB1 co-expressed with TLR4,and LOX-1 in infiltrated cells.Box A vs PBS treated C57 BL/6 mice had lower clinical scores and down-regulated corneal HMGB1,MIP-2,IL-1βexpression and neutrophil influx.Box B treatment amplified expression of MIP-2,IL-1β,TNF-α,HMGB1 and LOX-1 that induced by A.fumigatus in macrophage.Compared to the treatment of Box B only,the protein expression of IL-1β,TNF-αshowed inhibition of Box B combined with Poly(I),which also reduced the A.fumigatusevoked protein level of LOX-1 and phosphorylation level of p38-MAPK.The production of A.fumigatus-stimulated ROS was significantly declined after Box A pretreatment in PMN.CONCLUSION:Blocking HMGB1 reduces the disease response in C57 BL/6 mice.HMGB1 can amplify the host immune response through p38-MAPK,and is a target for treatment of A.fumigatus keratitis.

Anti-inflammatory effect of Heliotropium indicum Linn on lipopolysaccharide-induced uveitis in New Zealand white rabbits

AIM： To investigate the anti-inflammatory effect of an aqueous whole plant extract of Heliotropium indicum（HIE） on endotoxin-induced uveitis in New Zealand white rabbits.·METHODS： Clinical signs of uveitis including flares,iris hyperemia and miosis, were sought for and scored in1.0 mg/kg lipopolysaccharide（LPS）-induced uveitic rabbits treated orally with HIE（30-300 mg/kg）,prednisolone（30 mg/kg）, or normal saline（10 m L/kg）. The number of polymorphonuclear neutrophils infiltrating, the protein concentration, as well as levels of tumor necrosis factor-α（TNF-α）, prostaglandin E2（PGE2）, and monocyte chemmoattrant protein-1（MCP-1） in the aqueous humor after the various treatments were also determined. A histopathological study of the anterior uveal was performed.· RESULTS： The extract and prednisolone-treatment significantly reduced（P ≤0.001） both the clinical scores of inflammation（1.0-1.8 compared to 4.40 ±0.40 in the normal saline-treated rabbits） and inflammatory cells infiltration. The level of protein, and the concentrationsof TNF-α, PGE2 and MCP-1 in the aqueous humor were also significantly reduced（P ≤0.001）. Histopathological studies showed normal uveal morphology in the HIE and prednisolone-treated rabbits while normal saline-treated rabbits showed marked infiltration of inflammatory cells.· CONCLUSION： The HIE exhibits anti-inflammatory effect on LPS-induced uveitis possibly by reducing the production of pro-inflammatory mediators.

PMN apoptosis and its relationship with the lung injury after chest impact trauma

Background Polymorphonuclear neutrophil (PMN),one of the most important inflammatory cells,functions throughout the initiation,progression and resolution of inflammation. This study aimed at investigating the relationship between PMN apoptosis and the lung injury after chest impact trauma. Methods PMNs were purified from rabbits subjected to the chest impact trauma and their apoptosis,necrosis,survival and respiratory burst were detected by flow cytometry. Meanwhile,lactate dehydrogenase and (LDH) [Ca 2+ ]i were measured. Results The delayed apoptosis of PMNs in bronchoalveolar lavage fluid was observed from 2 hours to 12 hours after trauma,and viable cells increased. Respiratory burst of PMNs in bronchoalveolar lavage fluid was increased significantly from 2 hours with the peak at 8 hours. Meanwhile,lactate dehydrogenase in bronchoalveolar lavage fluid was higher than that in control ( P <0.05) from 4 hours to 24 hours,and intracellular free Ca 2+ in PMN was increased temporarilly. Conclusions Retention of PMN in tissues and the abnormality in apoptotic pathway inevitably generate persistent activation of PMN and excessive release of toxic substances,resulting in tissue injury. The temporary increase of intracellular free Ca 2+ may be responsible for the delayed apoptosis of PMN.