Characterization of an Organ Specific and Pathogen Responsive CC-NBS-LRR Gene from Cotton(Gossypium hirsutum L.)

Cotton diseases represent a major challenge to cotton growth.Cloning of a cotton pathogen response gene and promoter is of great importance to improve disease resistance.In this study,a

Cloning and analysing of 5' flanking region of Xenopus organizer gene noggin

Xenopus organizer specific gene Noggin possessesnearly all the characterestic properties of the action of or-ganizer to specify the embryonic body axis. To analyzehow the maternal inherited factors cofltrol its expressionpattern, we cloned the 5’ regulatory region of noggn gene.The 1.5 kb upstream sequense could direct reporter geneto express in vivo and data from deletion analysis indi-cated that a 229 base pair fragmet is essential for acti-vating noggn expression. We further demonstrated thatthe response elements within this regulatory region wereindeed under the control of growth factor activin and Wntsignaling pathway components.

Gastric autoimmune disorders in patients with chronic hepatitis C before,during and after interferon-alpha therapy

AIM:To explore the prevalence of autoimmune gastritis in chronic hepatitis C virus (HCV) patients and the influence of α-interferon (IFN) treatment on autoimmune gastritis. METHODS:We performed a prospective study on 189 patients with positive anti-HCV and viral RNA enrolled in a 12-month IFN protocol.We evaluated:a) the baseline prevalence of autoimmune gastritis,b) the impact of IFN treatment on development of biochemical signs of autoimmune gastritis (at 3,6 and 12 months),c) the evolution after IFN withdrawal (12 months) in terms of anti-gastric-parietal-cell antibodies (APCA),gastrin,anti-thyroid,and anti-non-organ- specific antibodies. RESULTS:APCA positivity and 3-fold gastrin levels were detected in 3 (1.6%) and 9 (5%) patients,respectively,at baseline,in 25 (13%) and 31 (16%) patients at the end of treatment (both P<0.001,vs baseline),and in 7 (4%) and 14 (7%) patients 12 months after withdrawal (P=0.002 and P=0.01 respectively,vs baseline;P=not significant vs end of treatment).The development of autoimmune gastritis was strictly associated with the presence of autoimmune thyroiditis (P=0.0001),no relationship was found with other markers of autoimmunity. CONCLUSION:In HCV patients,IFN frequently precipitates latent autoimmune gastritis,particularly in females.Following our 12-month protocol,the phenomenon generally regressed.Since APCA positivity and high gastrin levels are associated with the presence of antithyroid antibodies, development of autoimmune thyroiditis during IFN treatment may provide a surrogate preliminary indicator of possible autoimmune gastritis to limit the need for invasive examinations.

Endophytic Mycodiversity of Sacred Tree-Couroupita guianensis Aubl.

In the present study,a total of 142 fungal strains were isolated from 180 samples of the leaf,stem and bark tissues of sacred tree Couroupita guianensis from an unusual semi-arid tropical region.The tissues of the samples were grown in potato dextrose agar(PDA)medium and the endophytic mycoflora were identified based on the morphological characteristics.Among all the strains,130 were fertile,which belong to 11 species and 12 sterile morpho species.The relative frequency of isolated individual endophytic fungal group consists of hyphomycetes(41.5%),coelomycetes(32.4%),ascomycetes(13.4%),zygomycetes(4.2%)and sterile fungi(8.5%).Among the fungal isolates,Scytalidium acidophilum and Mycosphaerella sp.were recorded as the most dominant fungal isolates in the leaf tissues,Cladosporium cladosporioides and Colletotrichum falcatum were observed as the dominant endophytic fungal isolates in the stem tissues and Botryodiplodia theobromae was found to be predominant species in bark tissues of C.guianensis.The species of Colletotrichum were found only in stem tissues.Therefore,the endophytic mycodiversity were high and abundant and they were distinctly associated with host plant.All statistical analyses confirmed that leaf tissues contained more endophytes than the stem and bark while Colletotrichum exhibited organ specificity.The present study revealed that the sacred tree C.guianensis is one of the ecological niches for sheltering endophytic mycoflora eventhough in harsh climatic conditions.

Effect of cholecystokinin on cytokines during endotoxic shock in rats

AIM: To study the effect of cholecystokinin-octapeptide (CCK-8) on systemic hypotension and cytokine production in lipopolysaccharide (LPS)-induced endotoxic shock (ES) rats. METHODS: The changes of blood pressure were observed using physiological record instrument in four groups of rats: LPS (8mg.kg(-1),iv) induced ES; CCK-8 (40 microg.kg(-1), iv) pretreatment 10 min before LPS (8mg.kg(-1)); CCK-8 (40 micro.kg(-1), iv) or normal saline (control) groups. Differences in tissue and circulating specificity of the proinflammatory cytokines (TNF-alpha, IL-1beta and IL-6) were assayed with ELISA kits. RESULTS: CCK-8 reversed LPS-induced decrease of mean artery blood pressure (MABP) in rats. Compared with control, LPS elevated the serum level of IL-6 significantly (3567 +/- 687 ng.L(-1) vs 128 +/- 22 ng.L(-1), P【0.01), while contents of TNF-alpha and IL-1beta elevated significantly (277 +/- 86 ng.L(-1) vs not detectable and 43 +/- 9 ng.L(-1) vsnot detectable, P【0.01) but less extent than IL-6. CCK-8 significantly inhibited the LPS-induced increase in serum TNF-alpha IL-1beta and IL-6. LPS elevated spleen and lung content of IL-1beta significantly (5184 +/- 85 ng.L(-1) vs 1047 +/- 21 ng.L(-1) and 4050 +/- 614 ng.L(-1) vs not detectable, P【0.01), while levels of TNF-alpha and IL-6 also rose significantly but in less extent than IL-1beta. CCK-8 inhibited the LPS-induced increase of the cytokines in spleen and lung. In the heart, CCK-8 significantly inhibited LPS-induced increase of TNF-alpha (864 +/- 123 ng.L(-1) in CCK-8+LPS group vs 1599 +/- 227 ng.L(-1) in LPS group, P 【 0.01), and IL-1beta (282 +/- 93 ng.L(-1) in CCK-8+LPS group vs 621 +/- 145ng.L(-1) in LPS group, P 【 0.01). CONCLUSION: CCK-8 reverses ES, which may be related to its inhibitory effect on the overproduction of cytokines.

Get effective polyclonal antisera in one month

According to the traditional immunization procedure, after the first injection of the sample A (emulsion of aimed antigen and Freund's complete adjuvant) to immunize rabbit, successive injections of the sample B (emulsion of aimed antigen and Freund's incomplete adjuvant) were followed every 2-4 weeks. In general,high titer of the corresponding polyclonal antisera will be observed after 4-5 injections of sample B in 3-4months. This report presents a simply modified procedure that was able to stimulate the antisera formation in one month and achieve enough avidity to satisfy either Western blot or immunohistochemistry analysis.It just applied an additional injection of the sample A to the rabbit at the 3rd day after the primary immunization injection. You could gain the high titer of the antisera right after the first sample B injection in one month. This method has produced the desired results in three different recombinant antigens with different molecular weight (5.9 KD-55 KD) expressed from prokaryotic or eukaryotic cells.

Changes of CD8+CD28-T regulatory cells in rat model of colitis induced by 2,4-dinitrofluorobenzene

AIM:To determine the changes of CD8+ T subsets especially CD8+CD28-T regulatory cells in rat model of experimental colitis induced by 2,4-dinitrofluorobenzene (DNFB). METHODS:The rat model of experimental colitis was induced by enema with DNFB.Ten days later,colonic intraepithelial and splenic lymphooltes were isolated from colitis animals (n=16) and controls (n=8).The proportion of CD8+ T cells,CD8+CD28+ T cells and CD8+CD28-T regulatory cells were determined by flow cytometry. RESULTS:The model of experimental colitis was successfully established by DNFB that was demonstrated by bloody diarrhea,weight loss and colonic histopathology.The proportion of CD8+ T cells in either splenic or colonic intraepithelial lymphocytes was not significantly different between colitis animals and controls (spleen:34.6±7.24 % vs 33.5±9.41%, colon:14.0±8.93 % vs 18.0±4.06 %,P>0.05).But CD8+CD28- T regulatory cells from colitis animals were significantly more than those from controls (spleen:11.3±2.26 % vs 5.64±1.01%, colon:6.50±5.37 % vs 1.07±0.65 %,P<0.05).In contrast, CD8+CD28+ T cells from colitis animals were less than those from controls (spleen:23.3±6.14 % vs 27.8±9.70 %,P=0.06; colon:7.52±4.18 % vs 16.9±4.07 %,P<0.05).The proportion of CD8+CD28-T regulatory cells in splenic and colon intraepithelial CD8+ T cells from colitis animals was higher than that from controls (spleen:33.3±5.49 % vs 18.4±7.26 %, colon:46.0±14.3 % vs6.10±3.72 %,P<0.005). CONCLUSION:Experimental colitis of rats can be induced by DNFB with simplicity and good reproducibility.The proportion of CD8+CD28-T regulatory cells in rats with experimental colitis is increased,which may be associated with the pathogenesis of colitis.

Cholecystokinin octapeptide inhibits the in vitro expression of CD14 in rat pulmonary interstitial macrophages induced by lipopolysaccharide

OBJECTIVE: To study the effect of cholecystokinin octapeptide (CCK-8) on lipopolysaccharide (LPS)-stimulated pulmonary interstitial macrophages (PIM) in vitro. METHODS: PIM were isolated and cultured in the presence or absence of LPS, CCK-8, proglumide (the antagonist of CCK receptors) and vehicle. The expression of membrane CD14 (mCD14) protein was assayed by flow cytometry and soluble CD14 (sCD14) in the supernatant was analyzed semi-quantitatively by Western blot. TNF-alpha in the supernatant was detected with ELISA. RESULTS: CCK-8, at concentrations of 10(-7) mol/L and 10(-6) mol/L, significantly inhibited the expression of mCD14. Release of sCD14 and TNF-alpha in the supernatant was up-regulated by LPS (1 microg/ml) but reduced by CCK-8. The effect of CCK-8 was inhibited by proglumide. CONCLUSION: CCK-8 negatively modulated several functions of LPS-stimulated PIM through CCK receptors. This may be one of the mechanisms for CCK-8 to alleviate inflammation in lung tissue during endotoxemia.