ABSTRACT
This study aimed to investigate the effects of the house dust mite allergen Der p 1 on the secretion of tryptase from the human mast cell line HMC-1. Flow cytometry was used to determine the expression levels of protease-activated receptor-2 (PAR2) on the surface of HMC-1 cells. HMC-1 cells were treated with Der p 1, SLIGRL-NH2 (PAR2 agonist), LRGILS-NH2 (control peptide for PAR2), or Der p 1 + FSLLRY (PAR2 antagonist), and the tryptase levels were measured using enzyme-linked immunosorbent assay. The biological functions of PAR2 were determined using the calcium green indicator, and intracellular calcium fluorescence intensity in the different groups (Der p 1, SLIGRL-NH2, LRGILS- NH2, Der p 1 + FSLLRY, tryptase, tryptase + FSLLRY, or cell culture medium) was detected by laser scanning confocal microscopy. The mast cells expressed PAR2 receptor on their surfaces. Der p 1 alone induced a significant release of intracellular calcium and tryptase in HMC-1 cells compared with the SLIGRL- NH2 treatment group and the control group. The combination of Der p 1 and FSLLRY partly inhibited intracellular calcium and tryptase release in HMC-1 cells compared with the Der p 1 treatment group. Moreover, tryptase induced a significant release of intracellular calcium in the HMC-1 cells. Der p 1 induced HMC-1 cell degranulation and the release of tryptase by activating the PAR2 receptor on the cell surfaces. Tryptase activated the PAR2 receptor and induced intracellular calcium release from the HMC-1 cells in a positive feedback loop.
Subject(s)
Antigens, Dermatophagoides/pharmacology , Arthropod Proteins/pharmacology , Cysteine Endopeptidases/pharmacology , Mast Cells/enzymology , Tryptases/metabolism , Antigens, Dermatophagoides/immunology , Arthropod Proteins/immunology , Calcium/metabolism , Cell Line , Cysteine Endopeptidases/immunology , Flow Cytometry , Humans , Mast Cells/drug effects , Mast Cells/immunology , Oligopeptides/pharmacology , Receptor, PAR-2/antagonists & inhibitors , Receptor, PAR-2/biosynthesis , Receptor, PAR-2/immunology , Signal TransductionABSTRACT
The expression of interleukin-11 (IL-11) and its products STAT3 and phospho-STAT3 (p-STAT3) in patients with chronic superficial gastritis (CSG), chronic atrophic gastritis (CAG), and gastric cancer (GC) may provide insight into the diagnostic role of the IL-11/STAT3 signaling pathway in GC. Gastric mucosa specimens and serum samples were collected from 90 patients with CSG, CAG, and GC (30/group). The expression of IL-11, STAT3, and p-STAT3 was detected via immunohistochemistry and western blot. Additionally, serum levels of IL-11 were measured by enzyme-linked immunosorbent assay. For IL-11, 60% stained positive in CAG and 83.3% stained positive in GC, which were both higher than the value observed for CSG (33.3%). Moreover, the percent positive for IL-11 in GC was higher than that in CAG (P < 0.05). The percent positive for STAT3 in CAG (80%) and GC (83.3%) was higher than that in CSG (53.3%) (P < 0.05). Compared with CSG (36.7%), the percent positive for p-STAT3 in CAG (63.3%) and GC (86.7%) was also significantly higher. STAT3 expression was similar in GC and CAG, which was significantly higher than that in CSG. Expectedly, p-STAT3 expression gradually increased from CSG to CAG to GC. Furthermore, p-STAT3 levels were higher in GC tissues than in CAG (P < 0.01). Intriguingly, serum IL-11 levels gradually increased from CSG to CAG to GC, which coincided with disease severity. Together, these results suggest that the IL-11/STAT3 signaling pathway plays a critical role in human CAG, and may provide new targets to prevent and treat GC.
Subject(s)
Gastritis, Atrophic/genetics , Interleukin-11/biosynthesis , STAT3 Transcription Factor/biosynthesis , Stomach Neoplasms/genetics , Aged , Female , Gastric Mucosa/pathology , Gastritis, Atrophic/pathology , Humans , Interleukin-11/genetics , Male , Middle Aged , Precancerous Conditions/genetics , Precancerous Conditions/pathology , STAT3 Transcription Factor/genetics , Signal Transduction , Stomach Neoplasms/pathologyABSTRACT
Studies of genetic diversity and genetic population structure are critical for the conservation and management of endangered species. The Chinese sucker Myxocyprinus asiaticus is a vulnerable monotypic species in China, which is at a risk of decline owing to fluctuations in effective population size and other demographic and environmental factors. We screened 11 microsatellite loci in 214 individuals to assess genetic differentiation in both wild and cultured populations. The single extant wild population had a higher number of alleles (13) than the cultured populations (average 7.3). High levels of genetic diversity, expressed as observed and expected heterozygosity (HO = 0.771, HE = 0.748, respectively), were found in both wild and cultured populations. We also report significant differentiation among wild and cultured populations (global FST = 0.023, P < 0.001). Both STRUCTURE analysis and neighbor-joining tree revealed three moderately divergent primary genetic clusters: the wild Yangtze population and the Sichuan population were each identified as an individual cluster, with the remaining populations clustered together. Twenty-two samples collected from the Yangtze River were assigned to the cultured population, demonstrating the efficacy of artificial propagation to avoid drastic reduction in the population size of M. asiaticus. These genetic data support the endangered status of the M. asiaticus and have implications for conservation management planning.
Subject(s)
Cypriniformes/genetics , Genetic Speciation , Microsatellite Repeats , Polymorphism, Genetic , Animals , Cypriniformes/classification , Fisheries , Phylogeny , RiversABSTRACT
This study aimed to explore the law of superposition and masking between autoantibodies and alloantibodies, and to ensure the detection of alloantibodies and to improve the safety of warm autoimmune hemolytic anemia patients. Eight kinds of commercial IgG red blood cell antibody reagents were serially diluted, and 3 kinds of antibodies at dilutions showing a continuous gradual decline in agglutination strength with the corresponding antigen red blood cells were treated as the target antibodies. Anti-D and anti-M were treated as simulated autoantibodies, and anti-Fya was treated as a simulated alloantibody. Four concentrations, 4+, 3+, 2+ and 1+, of autoantibodies and three concentrations, 3+, 2+ and 1+, of alloantibodies were combined, and 12 kinds of hybrid antibodies were detected and evaluated by the anti-human globulin micro-column gel assay. When the simulated strong autoantibody (4+) was used, the alloantibodies (3+, 2+, 1+) had no effect on the final agglutination strength; when the strength of agglutination produced by the simulated autoantibody was less than 4+, and at the same time there were alloantibodies (3+, 2+, 1+), the differences in agglutination strength with a panel of RBCs could be clearly observed. Strong autoantibodies (4+) can exert a masking effect, leading to alloantibodies being undetected; autoantibodies less than 4+, will produce the superimposed effect with alloantibodies, resulting in differences in agglutination strength.
Subject(s)
Anemia, Hemolytic, Autoimmune/immunology , Autoantibodies/analysis , Isoantibodies/metabolism , Erythrocytes/immunology , Humans , Immunoglobulin G/immunologyABSTRACT
Several studies have documented the process of early embryonic development in poultry; however, the molecular mechanisms underlying its developmental regulation are poorly understood, particularly in ducks. In this study, we analyzed differential gene expression of embryos 6 and 25 h following oviposition to determine which genes regulate the early developmental stage in ducks. Among 216 randomly selected clones, 39 protein-encoding cDNAs that function in metabolism, transcription, transportation, proliferation/apoptosis, cell cycle, cell adhesion, and methylation were identified. Additionally, the full-length cDNA of the Nanog gene, encoding a 302-amino acid protein, was obtained. Quantitative real-time polymerase chain reaction analyses were performed to detect expression levels of the selected genes during early and late embryonic stages, which revealed that these genes are expressed in a particular spatial and temporal pattern. These results indicate that these genes may play pivotal roles in the process of area pellucida formation through a complex and precise regulatory network during development in duck embryos.