59 research outputs found
Gal3 expression in DRGs is increased after SNL.
<p>(A) The mRNA level of gal3 is increased in DRGs of rats subjected to L5 SNL. Total RNA was extracted from the fifth lumbar dorsal horn sections and was subjected to real-time PCR to analyze the relative expression level of gal3 in each sample. Each sample was analyzed in triplicate. The 2<sup>-ΔΔCt</sup> method was used to quantify the relative levels of gal3. β-actin was used as reference for mRNA. n = 10. *<i>p</i>< 0.05 vs sham, # <i>p</i>< 0.05 vs SNL group. (B and C) Western blot analysis of gal3 protein expression in the fifth lumbar dorsal horn sections in each sample.*<i>p</i>< 0.05 vs sham, # <i>p</i>< 0.05 vs SNL group.</p
MCP results in a decreased mechanicaland cold hypersensitivity.
<p>Mechanical (B) and cold (C) pain-related hypersensitivity developed after treatment with MCP (100 mg/kg/day) and Rapa (1 mg/kg/day) at the indicated time after surgery. n = 8. Data are expressed as mean ± SD. *<i>p</i>< 0.05 vs control, # <i>p</i>< 0.05 vs MCP group.</p
MCP inhibits SNL-induced activation of autophagy in spinal microglia.
<p>(A and B) Primary spinal microglial cells were isolatedfrom sham-, SNL-, and MCP-treated rats at day 10, and the autophagosome formation was visualized by assaying LC3 green puncta. Punctate staining is indicative for the redistribution of LC3 to autophagosomes. The average number of LC3 green puncta per cell with standard deviation for each group is presented. *<i>p</i>< 0.05 vs sham, # <i>p</i>< 0.05 vs SNL group. (C and D) Primary spinal microglial cells were isolated from sham-, SNL-, and MCP-treated rats at the indicated time, and LC3B protein levels were assayed using western blot analysis. <i>p</i>< 0.05 vs sham, # <i>p</i>< 0.05 vs SNL group. (E and F) Primary spinal microglial cells were isolated from sham-, SNL-, and MCP-treated rats at day 10, and the protein levels of p62 were assayed using western blot analysis. <i>p</i>< 0.05 vs sham, # <i>p</i>< 0.05 vs SNL group. (G-I) Microglial cells were isolated from sham-, SNL-, and MCP-treated rats at day 10, and the expression level of gal3 and autophagy activation was assayed using double-label immunofluorescence analysis. Scale bar = 50 μm.</p
Effect of ozone oxidative preconditioning on inflammation and oxidative stress injury in rat model of renal transplantation
<div><p>Abstract Purpose: To investigate the effect of ozone oxidative preconditioning (OzoneOP) on inflammation and oxidative stress injury in rat model of renal transplantation. Methods: Thirty six male Sprague Dawley (SD) rats were randomly divided into three groups. Sham group: rats were treated with opening and closing abdomen. Kidney transplantation group (KT group): SD rat received the donor’s left kidney derived from another SD rat. Ozone oxidative preconditioning and kidney transplantation (OOP+KT group): donor SD rats received OzoneOP treatments by transrectal insufflations before kidney transplantation. After transplantation, parameters of renal function of recipients were determined. Morphology and pathological changes of renal allograft were examined. Expression of NF-κBp65, HMGB-1 were also determined by Western-blot. Results: Compared to KT group, the morphology and pathological damages of renal allograft were less serious in OOP+KT group. Meanwhile, levels of SOD and GSH-Px of renal allograft in OOP+KT group were higher than those in KT group respectively. Western-blot showed that the expressions of NF-κBp65 and HMGB-1 in OOP+KT group were obviously less than those in KT group. Conclusion: Ozone oxidative preconditioning could attenuate the inflammatory reaction and oxidative stress injury in renal allograft, which might be related with the enhancement of anti-oxidative system and suppression of inflammatory reaction.</p></div
MCP inhibits LPS-induced activation of autophagy in microglia.
<p>(A and B) Mixed microglial cultures isolated from sham rats were treated with LPS (0.5ng/μl) and MCP (1 μg/μl), and the autophagosome formation was visualized by assaying LC3 green puncta. The average number of LC3 green puncta per cell with standard deviation for each group is presented. *<i>p</i>< 0.05 vs control, # <i>p</i>< 0.05 vs LPS group. Mixed microglial cultures isolated from sham rats were treated with LPS (0.5ng/μl) MCP (1 μg/μl) or Rapa (500 nM), and LC3B protein levels (C and D) or p62 protein levels (E and F) were assayed using western blot analysis. *<i>p</i>< 0.05 vs control, # <i>p</i>< 0.05 vs LPS group.</p
The auxotroph of the mutants can be satisfied by addition of the intermediate 2-KIC.
<p>Mycelial morphology of the wild-type PH-1, ΔFgLeu2A-10, ΔFgLeu2B-2, ΔFgLeu2AB-8 and ΔFgLeu2A-8C cultured on FGA medium amended with 2-KIC at different concentrations indicated in the figure at 25°C for 2 days.</p
<i>FgLEU2A</i> is required for DON biosynthesis of <i>F</i>. <i>graminearum</i>.
<p>The amounts of DON (mg/mg ergosterol) produced by the wild type strain PH-1, ΔFgLeu2A-10, ΔFgLeu2B-2, ΔFgLeu2AB-8 and ΔFgLeu2A-8C in infected wheat kernels and bars denote standard errors from three repeated experiments.</p
Relative expression of <i>FgLEU2A</i> and <i>FgLEU2B</i> in ΔFgLeu2A-10 and ΔFgLeu2B-2.
<p>The relative expression of <i>FgLEU2</i> genes is the relative amount of mRNA of each gene in the wild-type parent PH-1. Line bars in each column denote standard errors of three experiments.</p
Two <i>FgLEU2</i> Genes with Different Roles in Leucine Biosynthesis and Infection-Related Morphogenesis in <i>Fusarium graminearum</i>
<div><p>3-isopropylmalate dehydrogenase (IPMD) encoded by <i>LEU2</i> is a key enzyme in leucine (Leu) biosynthetic pathway. Analysis of the genome sequence of <i>Fusarium graminearum</i> revealed two paralogous <i>LEU2</i> genes (designated as <i>FgLEU2A</i> and <i>FgLEU2B</i>) in this fungus and the deduced amino acid sequences of FgLeu2A and FgLeu2B share 45% identity. Targeted disruption of individual <i>FgLEU2A/B</i> gene in <i>F</i>. <i>graminearum</i> assigned a more crucial role of FgLeu2A in Leu biosynthesis as disruption of <i>FgLEU2A</i> resulted in mutant (ΔFgLeu2A-10) that was Leu-auxotrophic and could not grow in minimal medium limited for amino acids, whereas <i>FgLEU2B</i> deletion mutant ΔFgLeu2B-2 was morphologically indistinguishable from the wild type strain PH-1. The growth defects of ΔFgLeu2A-10 could be overcome by exogenous addition of Leu at 0.25 mM. Double deletion of <i>FgLEU2A</i> and <i>FgLEU2B</i> (ΔFgLeu2AB-8) caused a more severe Leu-auxotrophic phenotype as the concentration of Leu exogenously added to medium to rescue the growth defect of ΔFgLeu2AB-8 should be raised to 1.25 mM, indicating a less important but nonnegligible role of FgLeu2B in Leu biosynthesis. Disturb of Leu biosynthesis caused by <i>FgLEU2A</i> deletion leads to slower growth rate, reduced aerial hyphal formation and red pigmentation on PDA plates and completely blocked conidial production and germination. All of the defects above could be overcome by Leu addition or complementation of the full-length <i>FgLEU2A</i> gene. ΔFgLeu2A-10 also showed significantly increased sensitivity to osmotic and oxidative stresses. Pathogenicity assay results showed that virulence of mutants lacking <i>FgLEU2A</i> were dramatically impaired on wheat heads and non-host cherry tomatoes. Additionally, a low level of deoxynivalenol (DON) production of ΔFgLeu2A-10 and ΔFgLeu2AB-8 in wheat kernels was also detected. Taken together, results of this study indicated a crucial role of FgLeu2A and a less important role of FgLeu2B in Leu biosynthesis and fungal infection-related morphogenesis in <i>F</i>. <i>graminearum</i> and FgLeu2A may serve as a potential target for novel antifungal development.</p></div
<i>FgLEU2A</i> is involved in adaptation to various cellular stresses in <i>F</i>. <i>graminearum</i>.
<p>Comparisons of mycelial inhibition percentages of each strain grown on YEPD medium amended with various cellular stresses at concentrations described in the figure and line bars in each column denote standard errors of three repeated experiments.</p
- …