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Objective · To prepare recombinant human TIGIT protein in E.coli and characterize its ability in binding Fusobacterium nucleatum (Fn). Methods · The gene of immunomodulatory protein of human TIGIT was amplified and cloned into pGEX4T2, and recombinant plasmid was transformed into E.coli BL21 (DE3) for GST-TIGIT fusion proteins were purified by the GST affinity chromatography and the interaction between GST-TIGIT fusion protein and Fn was tested by a pulldown assay. Results · Recombinant GST-TIGIT fusion protein expressed successfully in E.coli and was purified to homogeneity by GST affinity column. This protein could specifically bind to Fn, but not Lactobacillus acidophilus. Conclusion · High purify and activity of human GST-TIGIT fusion protein can be achieved by the prokaryotic expression system, and the adhesion between this protein and Fn has been preliminarily explored, which provides basis for further characterize interaction between them.
ABSTRACT
α1-Antitrypsin (α1-AT) belongs to serine protease inhibitor (Serpin) superfamily and is the main protease inhibitor in human circulation. It can inhibit many proteases to protect tissues from digradation. The mutant Z (Glu342Lys) of α1-AT predisposes to the early onset of emphysema due to decreased functional α1-AT in the lung and to neonatal hepatitis due to accumulation of α1-AT polymers in the endoplasmic reticulum of hepatocytes, which disrupts the balance between protease and protease inhibitors. This paper reviews recent research progress on the pathogenic mechanism and the prognosis of α1-antitrypsin deficiency.
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AimTo study the anti-inflammatory effect of astragalosides(AST) and its mechanisms of action. Methods The exudate volume, neutrophil count and protein content in exudate were measuredinthe carrageenan-induced air pounch model in rats. The content of PGE2 was assayed by radioimmunoassay,the activity of PLA2 by microacid titration assay, IL-8 by ELISA, and NO by nitrate reductase assay. The production of O2 in neutrophil was determined by cytochrome C assay. Results AST(40, 80 mg· kg -1) could markedly reduce the exudate volume, neutrophil count, protein content, the content of IL-8, and the production of O2.AST lowered PLA2 activity of neutrophil and accellular component in exudate, and it also decreased the contents of PGE2 and NO in exudate. Conclusion AST has an obvious anti-inflammatory effect on carrageenan-induced acute inflammation.Its mechanisms may be related to the inhibition of vascular permeability and leukocyte migration, as well as to the suppression of PLA2 activity and the reduction of IL-8, PGE2, NO and O2 production.
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2500mg/kg ( ig ) gave no any toxic symptom nor death. In long-term toxicity test, 50, 1000 and 2000mg/kg?d-1 ig on 30~90 consecutive days to rats and 280 and 560mg/kg?d-1 ig on 90 consecutive days to dogs respectively, revealed no significant toxicity in general physical signs, body weight, appearance. behavious, routine urine test, routine blood test, liver and kidney function tests, histological examinations of important organs, blood biochemistry tests and electrocardiogram, except that the platelet numbers of both animals increased.
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AIM To study the effect of alendronate sodium (Alen) on osteoclastic bone resorbing lacunae. METHOD Using the method of culturing osteoclasts on slices, the techniques of photomicrography microdensitometric scan and computer image analysis. RESULTS Alen(0 5,5 or 50 ?mol?L -1 )reduced the number and the surface area of osteoctastic bone resorbing lacunae dose dependently compared with model group there was significant defference ( P
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Effects of total glucosides of paeony ( TGP) on interleukin2 ( IL-2 ) production were studied with lL-2-dependent marine splenic lymohocytes. The results showed that IL-2 production by splenocytes was enhanced with low concentration of TGP (0.5-12.5 mg/L) , and diminished over the rang of 12.5-62.5mg/L. The concentration-effect curve seems to be bell-shaped. The data suggested that TGP hap two-sided regulatory effects on IL 2 production by splenocytes.
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AIM To study immunomodulation and antitumor activity of paeonol. METHODS Paeonol was administered singlely to HepA bearing mice. 14 d later, the inhibition of tumor weight were observed. IL-2 and TNF-? content in serum, IL-2 secretion by splencytes, TNF-? production by PM? were measured by RIA. RESULTS Paeonol possess markedly inhibitory activity on the growth of HepA tumor in mice, and IL-2 and TNF-? were induced signficantly. CONCLUSION Paeonol has the effects of antitumor. This effect may be derived from inducing IL-2 and TNF-? production.
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Method for measuring interleukin 1 ( IL -1 ) and effects of total glucosides of paeony(TGP) on IL -1 production were studied. The results showed that the best concentration for LPS -induced IL-1 production of M s in rats was 8mg/L, the best dilution for IL-1 of LPS-induced M s production was 1 : 90. TGP ( 0 .5 - 12 .5mg/L ) enhanced LPS-induced IL -1 production in a concentration dependent manner, but diminished in the higher concentration of TGP ( 62 .5~l25mg/L ) , the concentration-effect curve seems to be bell-shaped. The data suggested that TGP had two-sided regulatory effects on LPS -induced IL production of M s in rats.
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~3H-Clonidine, a potent and selective alpha2-adrenergic agonist was used to label alpha2-a-drenoceptors in intact lymphocytes isolated from rat spleen. Binding of ~3H-Clo-nidine was rapid(t1/2: 2min)and readily reversed by 10umol?L~(-1) clonidine(t1/2: 3-4min). ~3H-Clonidine saturationexperiments indicated a single c1ass of site with a K_D of 6.57?1.63nM and Bmax of 72.4?13.4 fmol/10~7 lymphocytes. Adrenergic agonists competed for ~3H-clonidine binding site with anorder of potency:epinephrine)norepinephrine)isoproterenol. These results show the presence ofalpha2-adrenoceptors in splenic lymphocytes. Computer analysis of competition experiments withadrenergic agonists revealed three classes of sites: high affinity site, medium affinity site and lowaffinity site. The affinity of high affinity site is 2-3 orders of magnitude higher than the one ofmedium affinity site, whereas the latter is the same orders higher than low affinity site Using im-proved Mishell-Dutton method, 10umol?L~(-1) clonidine suppressed the Ig M synthesis and thesuppression was blocked by 10 umol?L~(-1) phentolamine. These results indicate the suppression ofIg M antibody response to SRBC in vitro by clonidine is mediated by the alpha-adrenoceptors.