Project description:Clonorchiasis is associated with bile duct malignancy and the subsequent development of cholangiocarcinoma. Although this is likely caused by adult Clonorchis sinensis and its excretory-secretory products (ESP), the precise molecular mechanisms remain obscure. To evaluate the effect of C. sinensis infection on differential gene expression in host hepatocytes, we used cDNA microarrays in human cholangiocarcinoma cells through the mimicking of C. sinensis infestation, and analyzed differential mRNA expression patterns of host cells. Keywords: Time course
Project description:A total of 1,301 differentially expressed genes were identified: 521 upregulated and 780 downregulated. Furthermore, p21 and PAI levels decreased, which coincided with decreases in E-cadherin and increases in fibronectin expression, in cells stimulated with Clonorchis sinensis excretory/secretory products (ESPs) and NDMA. Cancer-related proteins, such as EGFR, K-ras, and PCNA, were significantly upregulated in cells exposed to C. sinensis ESPs and NDMA. Our results showed that C. sinensis ESPs and a small amount of NDMA were sufficient to induce cancerous transformation and epithelial-mesenchymal transition (EMT) in normal H69 cholangiocytes.
Project description:The excretory secretory products and positive sera of Clonorchis sinensis were collected for identifying the proteins presented in different infection periods, which specific to the sera of C. sinensis, but not combined with Fasciola hepatica, and Schistosoma japonicum. The White Japanese rabbits were artificially infected with C. sinensis, and the sera was collected at 7 days post-infection (dpi), 14 dpi, 35 dpi and 77 dpi. To identify the specific proteins to C. sinensis, Co-Immunoprecipitation (Co-IP) assay was used to pull down the sera root in C. sinensis, F. hepatica, and S. japonicum respectively by shotgun liquid chromatography tandem-mass spectrometry (LC-MS/MS).
Project description:In this study, Angiostrongylus cantonensis (AC), which parasitizes in the brain of the non-permissive host, such as mouse and human, is an etiologic agent of eosinophilic meningitis. Excretory-secretory (ES) products play an important role in the interaction between parasites and hosts’ immune responses. Inflammatory macrophages are responsible for eosinophilic meningitis induced by AC, and the soluble antigens of Angiostrongylus cantonensis fourth stage larva (AC L4), a mimic of dead AC L4, aggravate eosinophilic meningitis in AC-infected mice model via promoting alternative activation of macrophages. While whether AC L4 ES products, as well as its exosome-depleted excretory-secretory products (exofree) component play a role on macrophage activation remains unknown. In order to identify the AC L4 exofree signature regulating genes, BMDMs were treated with PBS (ctr), free (AC L4 exofree), IL-4, com (IL-4+AC L4 exofree) for 6h, 12h, and 24 h respectively and RNA-seq analysis was performed.
Project description:Clonorchis sinensis is a zoonotic parasite causing clonorchiasis associated with human diseases such as biliary calculi, cholecystitis, liver cirrhosis, and is classified as carcinogenic to humans for cholangiocarcinoma. MicroRNAs (miRNAs) are non-coding, regulating small RNA molecules essential for the complex life cycle of parasites and involved in parasitic infections. To identify and characterize miRNAs expressed in adult C. sinensis residing chronically in the biliary tract, we developed an integrative approach combining deep sequencing, bioinformatic predictions with stem-loop real-time PCR analysis. Here we report the use of this approach to identify and clone 6 new and 62,512 conserved C. sinensis miRNAs which belong to 284 families. There is strong bias on families, family members and sequence nucleotides in C. sinensis. Uracil is the dominant nucleotide, particularly at positions 1, 14 and 22, which were located approximately at the beginning, middle and the end of conserved miRNAs. There is no significant M-bM-^@M-^\seed regionM-bM-^@M-^] at the first and ninth positions commonly found in human, animals and plants. Categorization of conserved miRNAs indicated that miRNAs of C. sinensis are still innovated and concentrated along three branches of the phylogenetic tree leading to bilaterians, insects and coelomates. There are two miRNA strategies in C. sinensis for its parasitic life: keeping a large category of miRNA families of different animals and keeping a stringent conserved seed region with high active innovation in other place of miRNA mainly in the middle and the end, which are perfect for the parasite to perform its complex life style and for host changes. The present study represents the first large scale characterization of C. sinensis miRNAs, which have implications for understanding the complex biology of this zoonotic parasite, as well as the miRNA studies of other related species such as Opisthorchis felineus and O. viverrini of human and animal health significance. Analysis of miRNA profile in parasite of C. sinensis
Project description:Paragonimus kellicotti extravesicle data collected from in vitro excretory/secretory products and from cyst fluid from the lungs of infected Meriones unguiculatus.
Project description:Clonorchis sinensis is a zoonotic parasite causing clonorchiasis associated with human diseases such as biliary calculi, cholecystitis, liver cirrhosis, and is classified as carcinogenic to humans for cholangiocarcinoma. MicroRNAs (miRNAs) are non-coding, regulating small RNA molecules essential for the complex life cycle of parasites and involved in parasitic infections. To identify and characterize miRNAs expressed in adult C. sinensis residing chronically in the biliary tract, we developed an integrative approach combining deep sequencing, bioinformatic predictions with stem-loop real-time PCR analysis. Here we report the use of this approach to identify and clone 6 new and 62,512 conserved C. sinensis miRNAs which belong to 284 families. There is strong bias on families, family members and sequence nucleotides in C. sinensis. Uracil is the dominant nucleotide, particularly at positions 1, 14 and 22, which were located approximately at the beginning, middle and the end of conserved miRNAs. There is no significant “seed region” at the first and ninth positions commonly found in human, animals and plants. Categorization of conserved miRNAs indicated that miRNAs of C. sinensis are still innovated and concentrated along three branches of the phylogenetic tree leading to bilaterians, insects and coelomates. There are two miRNA strategies in C. sinensis for its parasitic life: keeping a large category of miRNA families of different animals and keeping a stringent conserved seed region with high active innovation in other place of miRNA mainly in the middle and the end, which are perfect for the parasite to perform its complex life style and for host changes. The present study represents the first large scale characterization of C. sinensis miRNAs, which have implications for understanding the complex biology of this zoonotic parasite, as well as the miRNA studies of other related species such as Opisthorchis felineus and O. viverrini of human and animal health significance.
Project description:We examined gene expression profiles (27028 genes) in the livers of Sprague-Dawley rats with no infection and at 2 and 4 weeks after Clonorchis sinensis infection using Whole Rat Genome Microarray 4x44K v3 (GPL14745, Agilent-028282)
Project description:Analysis of host response to the infected Clonorchis sinensis metacercariae and adult worm. The infected tissues evidenced altered expression of genes involved in systems such as immune response and cell cycle regulation, as compared with normal tissues.
