Hyaluronan (HA) was discovered over 60 years ago, but its metabolism and cellular interactions have only recently received detailed attention. HA is synthesized in the plasma membrane by addition of monosaccharides to the reducing terminal.
In tissues, it occurs bound to plasma membranes, aggregated with other macromolecules, or as free polysaccharide. Tissue HA enters the bloodstream in significant amounts through the lymph and is rapidly absorbed via a receptor into liver endothelial cells, where degradation follows. HA levels in serum are normally 10-100 ug/l, but can be elevated with aging especially over 70 years old, and in liver cirrhosis, rheumatoid arthritis and progressive systemic sclerosis, due either to impaired hepatic uptake or increased production.
Studies on urine, skin diseases, cancers and progeroid syndromes have identified other causes of deranged HA.metabolism. Recent biochemical and clinical analyses have indicated the anti-inflammatory activity of HA in several areas including rheumatoid arthritis and osteoarthritis by controlling cell proliferation, activation and chemotaxis.

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Leukocytes collected from the pouch fluid 4 h after the antigen challenge in the air pouch-type allergic inflammation model in rats produced proteinaceous chemotactic factors for neutrophils when they were incubated. The chemotactic factor production by the leukocytes from the immunized rats was significantly higher than that from the non-immunized rats. The production of the neutrophil chemotactic factors was inhibited by the nonspecific protein kinase inhibitor K-252a, the protein kinase C inhibitor H-7, and the tyrosine kinase inhibitor genistein, but was not inhibited by the cAMP-dependent protein kinase inhibitor H-89, suggesting that protein kinase C and tyrosine kinase but not cAMP-dependent protein kinase are activated in the leukocytes infiltrated into the inflammatory site to produce the neutrophil chemotactic factors.
Isoelectric focusing of the neutrophil chemotactic factors in the conditioned medium revealed that the leukocytes produce two factors, leukocyte derived neutrophil chemotactic factor (LDNCF) -1 and LDNCF-2, of which the pI values are 4-5 and >8, respectively. LDNCF-2, a major component of the neutrophil chemotactic factors, was purified by gel chromatography and reverse-phase HPLC, and the Nterminal amino acid sequence of the purified LDNCF-2 was determined.
It was demonstrated that the sequence (1 to 20) was identical to the 32 to 51 amino acid sequence of the proform of the rat macrophage inflammatory protein (MIP) -2. Furthermore, RT-PCR revealed that the level of MIP-2 mRNA was higher in the leukocytes from the immunized rats than that from the non-immunized rats.

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In the present study, we have found that unusual lymphocytes which phenotypically express Mac-1 (CD11b) exist in a large proportion (-20%) in murine lung. Further study revealed that these cells were T lymphocytes which expressed low intensity of CD3 and αβ T cell receptor. In addition, they express neither CD4 nor CD8, while conventional thymus-derived T lymphocytes usully have either of these molecules.
Mac-1⁺ lymphocytes accumulated in the lung and the expression of MHC class II and ICAM-1 on their surface was augmented after intravenous infection with Mycobacterium bovis BCG. Taken together, this population of lymphocytes might play some important role in the host defense against mycobacterial infection, although it remains to be elucidated whether they are double negative αβ T cells or the other population.

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Sairei-to (TJ114) is an extract with a multitude of constituents, prepared from 12 mixed raw herbs, i.e., Bupleurum Root, Alisma Rhizome, Pinellia Tuber, Scutellaria Root, Atractylodes Lancea Rhizome, Jujube Fruit, Chuling, Ginseng Root, Hoelen, Glycyrrhiza Root, Cinnamon Bark and Ginger Rhizome. The pharmacological actions of TJ114 include promotion of endogenous steroid secretion and enhancement of steroid effects. Type II collagen induced arthritis was generated in DBA/ 1J mice. Mice were immunized with a series of subcutaneous injections of type II collagen with complete Freund's adjuvant. Three weeks later, incomplete Freund's adjuvant and an equal quantity of type II collagen were injected at the same site as a booster, thereby creating a mouse with type II collagen induced arthritis.
TJ114 was effective in delaying and mitigating the development of type II collagen-induced arthritis after both the intial immunization and the booster immunization. As regards the optimal dosage of TJ114, 500 mg/kg/day appeared most effective with respect to mitigation of swelling in the extremities as well as improvement in the inflammation score. The latter is a severity of bone lesions. The special characteristic which emerged in the present study was the significant inhibitory effect of TJ114 with respect to the blast transformation of splenic cells in response to type II collagen as well as inhibition of anti-type II collagen antibody production in the blood serum.

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There have been increasing findings indicating that free radicals and microcirculatory disturbances in gastric mucosa play an important role in the mucosal injury induced by non-steroidal antiinflammatory drugs, besides the inhibitory effect of these drugs on prostaglandin production. Therefore, we evaluated the effect of ampiroxicam, a prodrug of piroxicam, on the levels of reduced glutathione (GSH) in rat gastric mucosa and the mucosal microcirculation in rats by using a technique of intravitalmicroscopy. GSH levels were examined 5 hr after the oral administration of ampiroxicam, piroxicam and indomethacin. Ampiroxicam (13.5 mg/kg) caused a significant decrease of the gastric mucosal GSH content, however, this effect was apparently weaker than those of piroxicam (10 mg/kg) and indomethacin (12.5 mg/kg) . In observation of the basal part of the mucosal microcirculation by intravitalmicroscopy, local application of ampiroxicam (0.2 mM) on gastric submucosa had little or no effect on the diameter of collecting venules in the mucosa. On the other hand, a similar application of piroxicam or indomethacin produced a long-lasting constriction of the venules.
These results suggest that a weak effect of ampiroxicam on the microcirculation system in gastric mucosa may be one explanation of the ameliorated ulcerogenic activity of this drug.

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Murine sympathetic superior cervical ganglia were co-cultured with mesenteric lymph node explants isolated from nematode Nippostrongylus brasiliensis infected mice (Nb-MLN) to examine if inflammatory cytokine response affects the ability of MLN to induce neurite outgrowth. Explants were cultured with ganglia at a distance of 1 mm in a Matrigel layer, and number of neurites towards explants were counted at 1 and 2 days in culture. While uninfected MLN exerted a smaller effect on neurite outgrowth, Nb-MLN promoted the extent of the outgrowth, and the maximum response was observed 10 days after infection. To demonstrate the roles of cytokines in the promotion of the neurite outgrowth, various anti-cytokines were added to the co-cultures. Uninfected MLN-induced neurite outgrowth was inhibited by antinerve growth factor (NGF) and anti-IL-1β. On the other hand, Nb-MLN-induced outgrowth was inhibited not only by anti-NGF and anti-IL-1β but also by anti-IL-3, anti-IL-6 and anti-GM-CSF. Since IL-1 is known to stimulate NGF production, IL -1 might be necessary for NGF-mediated neurite outgrowth in lymph nodes. Moreover, IL-3, IL-6 and GM-CSF produced during inflammation might also play important roles in the stimulation of nerve growth in vivo.

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In order to clarify the antinephritic mechanisms of acteoside (TJC-160), a component of Stachys sieboldii MIQ (Chorogi), the effects of TJC-160 on adhesion molecule were evaluated with crescentic-type anti-GBM nephritic rats. TJC-160 was administered orally, once a day from the day after the injection of anti-GBM serum and continued througout the experiment. TJC-160 improved proteinuria during the experimental period. The number of LEA-1 positive cells or ICAM-1 expression in glomeruli of nephritic rats increased by day 20 or day 25, respectively. In addition, the number of ED 1, CD8, RT 7.2 (granulocytes) and Ia positive cells in glomeruli of nephritic rats was elevated by the 15th day, and then restored to the normal level by the 35th days. TJC-160 markedly prevented the increased number of LFA-1 and the expression of ICAM-1 in glomeruli. Moreover, TJC-160 suppressed the increase in the number of ED-1, CD8 and Ia positive cells by 40-60%. TJC-160, however, failed to suppress the production of specific antibody against rabbit γ globulin. These results indicate that the antinephritic effect of TJC-160 may be at least partly due to the inhibition of glomerular infiltration of certain leukocyte subsets and the expression of adhesion molecules.

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We examined the changes in vascular permeability induced by inflammatory mediators in mouse skin by the Pontamine sky blue (PSB) leakage technique. Five min after i.v. administration of PSB, mediators or saline were injected (0.1 ml/site, s.c.) into the back. Sixty min later, the dye accumulated in the skin was determined colorimetrically. Plateletactivating factor (PAF, 45-180 pmol/site), 5-hydroxy-tryptamine (5-HT, 120-960 pmol/site), substance P (100-1, 000 pmol/site) and histamine (550-5, 500 pmol/site) produced dose-related increases in vascular permeability. The dose response curve induced by bradykinin (20-80 nmol/site) was bellshaped. Pretreatment with indomethacin significantly reduced the increase in vascular permeability elicited by the 5 substances tested. Coadministration of prostaglandin (PG) E₂ partially reversed the inhibitory effect of indomethacin on the PAF- or histamine induced-increase in vascular permeability. Increases in vascular permeability induced by 5-HT, substance P and bradykinin were inhibited by N^G-nitro-l-arginine methyl ester (L-NAME) and methylene blue, but not by D-NAME ; however, PAF-and histamine-induced increases in vascular permeability were not affected by L-NAME.
These results suggest that eicosanoids may play a role in the effect of all 5 substances tested, and especially the increases in plasma extravasation induced by PAF and histamine are mediated by the production of PGE₂, and that NO may play a role, at least partly, in the case of 5-HT, substance P and bradykinin.

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The myeloperoxidase (MPO) activity of polymorphonuclear leukocytes (PMNs) during phagocytosis was investigated using a pro-chemiluminescence probe tyrosine. The tyrosine dependent chemiluminescence (T-CL) appeared when PMNs was stimulated with opsonized zymosan (OZ) and enhanced by the addition of tyrosine.
This method can measure the released MPO activity in intact PMNs over time.

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Rubus suavissimus is a kind of the plants in Rosaceae, the leaves of which have been brewed and drunk as a usual drink in China. It has been experientially known that the tea is effective for sore throat and laryngitis.
In the present experiments, we investigated the influence of the Rubus suavissimus extracts on experimental allergy. As the results, the extracts showed the suppressive effects on cyclooxygenase activity and anaphylactically induced histamine release from rats peritoneal mast cells, in vitro. In addition to these, oral administration of the extracts significantly inhibited either skin reaction induced by compound 48/80 or passive cutaneous anaphylaxis in the rat.
These results suggest that the rubus suavissimus extracts are therapeutically effective for inflammatory and allergic diseases.

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Recently some reports say that nitric oxide (NO) increased in exhaled air of asthmatic children. We measured NO concentration of asthmatic children and control subjects.
The result was that NO was significantly higher in exhaled air of asthmatic children than control subjects. There was no significant difference between asthmatic children who had athmatic attacks within 1 month and those who did not. But the children who suffered attacks within 2 weeks showed higher NO concentration than those who did not.
We assume that measurement of NO concentration in exhaled air is efficient on judgement of asthmatic state.

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