The regulatory roles of Fasciola hepatica GSTO1 protein in inflammatory cytokine expression and apoptosis in murine macrophages.

Fascioliasis, a global zoonotic parasitic disease, is mainly caused by Fasciola hepatica (F. hepatica) parasitizing in the livers of hosts, mainly humans and herbivores. Glutathione S-transferase (GST) is one of the important excretory- secretory products (ESPs) from F. hepatica, however, the regulatory roles of its Omega subtype in the immunomodulatory effects remain unknown. Here, we expressed F. hepatica recombinant GSTO1 protein (rGSTO1) in Pichia pastoris and analyzed its antioxidant properties. Then, the interaction between F. hepatica rGSTO1 and RAW264.7 macrophages and its effects on inflammatory responses and cell apoptosis were further explored. The results revealed that GSTO1 of F. hepatica owned the potent ability to resist oxidative stress. F. hepatica rGSTO1 could interact with RAW264.7 macrophages and inhibit its cell viability, furthermore, it may suppress the production of pro-inflammatory cytokines IL-1ß, IL-6 and TNF-α, but promote the expression of anti-inflammatory cytokine IL-10. In addition, F. hepatica rGSTO1 may down-regulate the ratio of Bcl-2/Bax, and increase the expression of pro-apoptotic protein caspase-3, thereby eliciting the apoptosis of macrophages. Notably, F. hepatica rGSTO1 inhibited the activation of nuclear factor-κB (NF-κB) and mitogen­activated protein kinases (MAPKs p38, ERK and JNK) pathways in LPS-activated RAW264.7 cells, exerting potent modulatory effects on macrophages. These findings suggested that F. hepatica GSTO1 can modulate the host immune response, which provided new insights into the immune evasion mechanism of F. hepatica infection in host.

Molecular Characteristics and Genetic Diversity of Fasciola Hepatica from Sheep in Xinjiang, China.

Introduction: Fasciola hepatica is a trematode infecting ruminants worldwide and occasionally affecting other animal species, including humans. It causes significant economic losses. Geographic distribution and patterns of infection must be considered before control and management measures are developed for this parasite. DNA molecular markers are useful for the identification of flukes and elucidation of their genetic evolution. Therefore, the population structure of F. hepatica was studied using this method in sheep in Xinjiang, China. Material and Methods: The molecular characteristics, genetic relationships within the population and dispersal patterns of F. hepatica isolates were analysed based on the cox1 and nad1 genes. The population structure of F. hepatica from three regions of Xinjiang was explored and a neutrality test was conducted. Results: The cox1 and nad1 genes have 21 and 42 variable sites, respectively, which can be classified into 34 and 33 haplotypes. Median-joining network and phylogenetic tree analyses showed that there was no significant variation in F. hepatica isolates between the three geographical regions. Analysis of variance revealed that the genetic variation of F. hepatica was mainly present within the populations. The neutrality test indicated that the populations were relatively stable but the Hami population may have undergone short-term expansion. Conclusion: This study revealed for the first time the molecular characteristics, genetic diversity and dispersal patterns of F. hepatica isolates from sheep in Xinjiang, thus providing new insights into the genetic variation and haplotype diversity of F. hepatica from indigenous sheep.

Molecular Characteristics and Potent Immunomodulatory Activity of Fasciola hepatica Cystatin

Cystatin, a cysteine protease inhibitor found in many parasites, plays important roles in immune evasion. This study analyzed the molecular characteristics of a cystatin from Fasciola hepatica (FhCystatin) and expressed recombinant FhCystatin (rFhcystatin) to investigate the immune modulatory effects on lipopolysaccharide-induced proliferation, migration, cytokine secretion, nitric oxide (NO) production, and apoptosis in mouse macrophages. The FhCystatin gene encoded 116 amino acids and contained a conserved cystatin-like domain. rFhCystatin significantly inhibited the activity of cathepsin B. rFhCystatin bound to the surface of mouse RAW264.7 cells, significantly inhibited cell proliferation and promoted apoptosis. Moreover, rFhCystatin inhibited the expression of cellular nitric oxide, interleukin-6, and tumor necrosis factor-α, and promoted the expression of transforming growth factor-β and interleukin-10. These results showed that FhCystatin played an important role in regulating the activity of mouse macrophages. Our findings provide new insights into mechanisms underlying the immune evasion and contribute to the exploration of potential targets for the development of new drug to control F. hepatica infection.

Occurrence of Gastrointestinal Parasites in Camels in the Tianshan Mountains Pastoral Area in China.

INTRODUCTION: Gastrointestinal parasites are some of the most common pathogens which are seriously harmful to the camel's health. The infection status of gastrointestinal parasites in camels (Camelus bactrianus) in the Tianshan Mountains pastoral area in China is still unclear. The aim of this study was to investigate the species and infection intensity of gastrointestinal tract parasites in local camels. MATERIAL AND METHODS: A total of 362 fresh faecal samples were collected and examined for parasite eggs using the saturated saline floating and natural sedimentation method. The parasite eggs were subjected to morphological and molecular examination and identification, and the infection rate and mean intensity of the parasites were analysed. RESULTS: A total of 15 gastrointestinal tract parasite species' eggs were identified, with a detection rate of 100%. Ostertagia spp. (100%) and Trichostrongylus spp. (98.1%) were dominant. Camels were often coinfected by 5-14 species. The average number of eggs per gram of faeces was higher for Ostertagia spp. (298), Haemonchus contortus (176) and Nematodirus spp. (138). The number of species of parasites infecting young camels was significantly lower than that of adult camels, but the infection intensity in young camels was significantly higher. CONCLUSION: Gastrointestinal parasites were highly prevalent in camels from the Tianshan Mountains pastoral area in China. This finding provides important epidemiological data for the prevention and control of associated infections in camels.

Molecular Detection and Genetic Diversity of porcine Circovirus Type 3 in Commercial Pig Farms in Xinjiang Province, China.

INTRODUCTION: Porcine circovirus type 3 (PCV3) is a newly discovered porcine circovirus. The molecular characteristics and genetic evolution of PCV3 in Xinjiang province, China still being unclear, the aim of the study was their elucidation. MATERIAL AND METHODS: A total of 393 clinical samples were collected from pigs on commercial farms in nine different regions of Xinjiang and phylogenetic analysis based on full-length Cap genes was performed. RESULTS: The prevalence at farm level was 100%, while in all the tested samples it was 22.39%. Nine PCV3 strains were detected in Xinjiang province and they shared 98.9-99.3% nucleotide and 97.5-100.0% Cap gene amino acid sequence identities with other epidemic strains from China and abroad. Compared with other epidemic strains of PCV3, there were 26 base mutation sites in the Cap gene in the nine Xinjiang strains, resulting in the mutation of amino acids at positions 20, 24, 75, 77, 108, 111 and 206. Phylogenetic analysis showed that these strains can be divided into two different genetic groups, to the first of which five strains affiliated and divided between subgroups 1.1 and 1.2, and to the second of which the other four strains affiliated and similarly divided between subgroups 2.1 and 2.2. CONCLUSION: PCV3 circulates widely among commercial pig farms in Xinjiang province, China, and displays obvious genetic diversity. The results provide epidemiological information useful for the prevention and control of PCV3 infection in the pig industry.

Sequential therapy to hepatic VX_2 tumor of rabbit with radiofrequency ablation combined with transcatheter arterial chemoembolization and external high frequency thermotherapy / 重庆医科大学学报

Objectives:To evaluate the effect of hepatic VX2 tumor mode treated by radiofrequency ablation(RFA),external high frequency thermotherapy(HFT) and transcatheter arterial chemoembolization(TACE).Methods:we replicated the model of hepatic tumor via transplantation and treated it with RFA,TACE and HFT.All the experimental rabbits was divided into four groups based on the different combination:group A:RFA+TACE+HFT,group B:RFA+HFT,group C:FA+TACE,Group D:TACE+HFT.Results:(1)After treatment,ALT and AST rised obviously on the first day in all groups,especially group A,which recovered the most slowly.In group B,however,ALT and AST was the lowest on the first day but decreased to normal most rapidly(P