Project description:Cholesterol gallstone is a common disease, but the pathogenesis of gallstone remains a mystery. The pathogenetic factors of gallstone disease include a genetic background, cholesterol condensation and bile supersaturation, while the risk factors may include fatty liver, diabetes, obesity and microbiome dysbiosis. Dysbiosis refers to altered bacterial composition and abundance, which is found to be associated with the pathogenesis of variety of diseases. Mass spectrometry-based proteomics emerges as a powerful tool to identify protein expressions of human samples. A number of proteomic studies have been performed to analyze human protein contents of bile samples. All these studies focus on the analysis of human proteins, and no study to date has been performed to analyze the protein expression of microbiome in bile samples associated with gallstone disease. In current study, we performed a label-free metaproteomic analysis of gallbladder bile samples from cholesterol gallstone disease patients and normal individuals. Maxquant was used in the comprehensive database searching to infer protein groups with quantitation. We attempt to investigate the dysbiosis changes, potential virulence factor of microbiome and host immune responses which contribute to gallstone formation and cholelithiasis.
Project description:Cholesterol gallstone is a common disease, but the pathogenesis of gallstone remains a mystery. The pathogenetic factors of gallstone disease include a genetic background, cholesterol condensation and bile supersaturation, while the risk factors may include fatty liver, diabetes, obesity and microbiome dysbiosis. Dysbiosis refers to altered bacterial composition and abundance, which is found to be associated with the pathogenesis of variety of diseases. Mass spectrometry-based proteomics emerges as a powerful tool to identify protein expressions of human samples. A number of proteomic studies have been performed to analyze human protein contents of bile samples. All these studies focus on the analysis of human proteins, and no study to date has been performed to analyze the protein expression of microbiome in bile samples associated with gallstone disease. In current study, we performed a label-free metaproteomic analysis of gallbladder bile samples from cholesterol gallstone disease patients and normal individuals. ProteoStorm was used for the first-step comprehensive database searching to identify microbial and host peptides, and Maxquant was used in the second-step refined database searching to infer protein groups with quantitation. We attempt to investigate the dysbiosis changes, potential virulence factor of microbiome and host immune responses which contribute to gallstone formation and cholelithiasis.
Project description:Epigenome-wide methylation levels were measured in patients from Chiel with gallstone disease, gallbladder dyplasia or gallbladder cancer using Illumina Infinium methylation arrays.
Project description:Objectives: Obstructive Sleep Apnea (OSA) is related to repeated upper airway collapse, intermittent hypoxia, and intestinal barrier dysfunction. The resulting damage to the intestinal barrier may affect or be affected by the intestinal microbiota. Methods: A prospective case-control was used, including 48 subjects from Sleep Medicine Center of Nanfang Hospital. Sleep apnea was diagnosed by overnight polysomnography. Fecal samples and blood samples were collected from subjects to detect intestinal microbiome composition (by 16S rDNA gene amplification and sequencing) and intestinal barrier biomarkers – intestinal fatty acid-binding protein (I-FABP) and D-lactic acid (D-LA) (by ELISA and colorimetry, respectively). Results: The severity of OSA was related to differences in the structure and composition of the intestinal microbiome. Enriched Fusobacterium, Megamonasa, Lachnospiraceae_UCG_006, and reduced Anaerostipes was found in patients with severe OSA. Enriched Ruminococcus_2, Lachnoclostridium, Lachnospiraceae_UCG_006, and Alloprevotella was found in patients with high intestinal barrier biomarkers. Lachnoclostridium and Lachnospiraceae_UCG_006 were the common dominant bacteria of OSA and intestinal barrier damage. Fusobacterium and Peptoclostridium was independently associated with apnea-hypopnea index (AHI). The dominant genera of severe OSA were also related to glucose, lipid, neutrophils, monocytes and BMI. Network analysis identified links between the intestinal microbiome, intestinal barrier biomarkers, and AHI. Conclusions: The study confirms that changes in the intestinal microbiota are related to intestinal barrier biomarkers among patients in OSA. These changes may play a pathophysiological role in the systemic inflammation and metabolic comorbidities associated with OSA, leading to multi-organ morbidity of OSA.
Project description:Following a large-scale genome-wide association study of gallstone disease, we performed RNA sequencing from tissues of four human gallbladders (3 healthy controls and 1 case with chronic gallstones) and one liver sample from the gallstone case. We aimed to determine the expression patterns of gallstone disease-associated genes in gallbladder and liver, two organs of interest in disease etiology.
Project description:A human gut-on-a-chip microdevice was used to coculture multiple commensal microbes in contact with living human intestinal epithelial cells for more than a week in vitro and to analyze how gut microbiome, inflammatory cells, and peristalsis-associated mechanical deformations independently contribute to intestinal bacterial overgrowth and inflammation. This in vitro model replicated results from past animal and human studies, including demonstration that probiotic and antibiotic therapies can suppress villus injury induced by pathogenic bacteria. By ceasing peristalsis-like motions while maintaining luminal flow, lack of epithelial deformation was shown to trigger bacterial overgrowth similar to that observed in patients with ileus and inflammatory bowel disease. Analysis of intestinal inflammation on-chip revealed that immune cells and lipopolysaccharide endotoxin together stimulate epithelial cells to produce four proinflammatory cytokines (IL-8, IL-6, IL-1β, and TNF-α) that are necessary and sufficient to induce villus injury and compromise intestinal barrier function. Thus, this human gut-on-a-chip can be used to analyze contributions of microbiome to intestinal pathophysiology and dissect disease mechanisms in a controlled manner that is not possible using existing in vitro systems or animal models. 6 samples, 2 biological replicates for each 3 conditions.
Project description:Graft-versus-host disease (GvHD) is critical complication after allogeneic hematopoietic stem cell transplantation (HSCT). The immunosuppressants given to patients undergoing allogeneic HSCT disturb the microbiome and the host immune system, potentially leading to dysbiosis and inflammation. The intestinal microbiome is a target for the development of novel therapies for GvHD. We determined the effect of the combination of tacrolimus (FK506) and Lactobacillus acidophilus on GvHD.