Granulosus Cysts Research Articles

HLA class II profile in patients with different stages of cystic echinococcosis according to the WHO ultrasound imaging classification.

The factors involving in the natural history and determinants of different features of human cystic echinococcosis (CE) are not adequately understood. Several host-related factors including the genetic structure of the host and human leukocyte antigens (HLAs) are believed to be involved in the natural history of CE in humans. The present study was conducted to investigate the association between HLA class II genes and active and inactive stages of hepatic cystic echinococcosis. Echinococcus granulosus cyst samples and patient information were collected from the biobank of the Iranian Hydatid Disease Registry from 2019 to 2022. HLA-DRB and HLA-DQB were characterized by PCR method. CE patients were categorized into three active (CE1 and CE2), inactive (CE4 and CE5), and transitional (CE3) stages according to the WHO ultrasound classification of CE. In total, 77 participants including 38 patients (36.8% men and 63.2% women) with different stages of CE as well as 39 healthy individuals (38.5% men and 61.5% women) were included in the study. Findings of the study showed that the frequency of HLA-DRB1*03 was significantly lower in the patients compared to the healthy individuals. The frequencies of HLA-DQB and HLA-DRB alleles were not differed significantly between active, inactive, and transitional stages of E. granulosus cysts. Findings of this study indicate the potential role of this allele in the susceptibility of human to cystic echinococcosis. Further large-scale studies in different endemic countries are required to document the significance of HLA-DQB and HLA-DRB as a host-related factor in the natural history of CE in human.

Study on the genotypes of Echinococcus granulosus in yaks and sheep from Langkazi County in Tibet Autonomous Region of China based on mitochondrial cox1 and nad1.

To determine the genotypes of the epidemic strains of Echinococcus granulosus in livestock in Tibet, samples of E. granulosus cysts were collected from 11 yaks and 62 sheep. Genomic DNA was extracted from these samples, and gene fragments of mitochondrial cytochrome c oxidase subunit I (cox1) and NADH dehydrogenase subunit I (nad1) were amplified by PCR and sequenced. DNASTAR and MAGA7.0 were employed for homology analysis and phylogenetic tree construction. Echinococcus granulosus cysts were detected in 56.2% (41/73) of the samples screened. Of these, 63.4% (26/41) were identified as E. granulosus G1 genotype (common sheep strain), 24.4% (10 /41) as G3 genotype (buffalo strain), and 12.2% (5/41) were G6 genotype (camel strain). The study concludes that yaks and sheep in Langkazi county, Tibet, carry three E. granulosus genotypes (G1, G3, and G6), with the G1 genotype the predominant genotype in the region. This study clarifies the distribution of E. granulosus genotypes, providing genetic data and insight for the surveillance and prevention of echinococcosis.

Toxicity of geranium oil, geraniol and their nanoemulsions on protoscoleces of hydatid cyst under in vitro conditions

Surgery is the main efficacious treatment for many cases of cystic echinococcosis by removing Echinococcus granulosus cysts. However, to reduce risk of cyst spillage and insemination of the content, using a scolicidal agent is crucial. Considering side effects of available scolicidals, and growing nano-technological approaches in novel pharmaceuticals, the present study aimed to find out the scolicidal activity of geraniol (GL), geranium oil (GM), and their developed nanoemulsions (Nano-GL and Nano-GM) on the protoscoleces of E. granulosus. Nanoemulsions were developed by ultrasonication emulsification and characterised by dynamic light scattering method (DLS). The scolicidal effect of GL, GM, Nano-GL, and Nano-GM at different concentrations of 1, 2.5, 5, 10, 25, and 50 µg/mL were determined after 0.5, 1, and 2 hours of incubation. Mortality rates were measured by eosin exclusion test. The average droplet size for Nano-GM and Nano-GL were 124.8 and 88.59 nm, respectively. After one hour of exposure, all tested concentrations of GL and Nano-GL resulted in higher than 90% mortality rates, while GM and Nano-GM killed 70.66% and 90.33% of protoscoleces, respectively. Based on 50% lethal concentration, Nano-GL was significantly more potent than Nano-GM (LC50: 4.52 vs 102.95 μg/mL). Results of scanning electron microscopy revealed tegumental disruption in the treated protoscoleces. This study described an easily applicable and eco-friendly procedure of nano-formulating functional phytochemicals, showing promising scolicidal activity in vitro. The developed formulations, especially Nano-GL, showed the characteristics of an ideal scolicidal agent. Further studies are needed to assess in vivo efficacy and safety of this formulation

Macrophage-mediated mechanisms of lung injury in the sensitization reaction to Echinococcus granulosus.

In this study, the impact of inhibiting the PI3K/AKT/NF-κB pathway on lung oxidative damage induced by Echinococcus granulosus cyst fluid was investigated. Twenty-four mice were randomly assigned to four groups. Three months after inoculation with hydatid cyst segments, mice in group A were treated with intraperitoneal and intratracheal saline injections; mice in group B were administered a caudal vein injection of a PI3K inhibitor, followed by cyst fluid sensitization; mice in group C received an AKT inhibitor via caudal vein, followed by cyst fluid sensitization; and mice in group D were subjected to cyst fluid sensitization without any inhibitor treatment. Cellular changes in lung tissues across all groups were evaluated, including pathological section analysis. Analysis of pulmonary tissue and serum from these mice included the assessment of PI3K/AKT/NF-κB pathway proteins, inflammatory factors, and related mRNA levels. Mice in groups B and C exhibited a higher proportion of M2-type macrophages and significantly lower levels of PI3K/AKT/NF-κB pathway proteins, inflammatory factors (interleukin-6 [IL-6]/tumor necrosis factor-α [TNF-α]), and oxidative markers in lung tissues compared to mice in group D (P < 0.05). Our results in this study indicate that activation of the PI3K/AKT/NF-κB pathway contributed to an increase in the M1 macrophage phenotype, leading to enhanced secretion of peroxidases and inflammatory factors. This mechanism plays a crucial role in the oxidative and inflammatory lung damage associated with allergic reactions to E. granulosus cyst fluid.

Epidemiology, Molecular, and Phylogenetic Characterization of Echinococcus granulosus Cysts in Slaughtered Farm Animals in Al-Jouf Province, Saudi Arabia

Echinococcosis, or hydatidosis, is a serious veterinary disease and public health issue worldwide, particularly in rural areas in which dogs have frequent contact with local herbivores. This study assessed the frequency of hydatidosis found among farm animals slaughtered in Al-Jouf Province in northern Saudi Arabia in 2021. A total of 156754 sheep, 36337 goats, 8590 camels, and 986 cattle were inspected for hydatidosis infection by comprehensive evaluation involving meticulous visual inspection and manual exploration of the internal organs through palpation. The cysts were subjected to molecular and phylogenetic analysis. The overall prevalence rates of hydatid cysts were 0.43%, 0.19%, 0.54%, and 0.51% in the inspected sheep, goats, camels, and cattle, respectively. The highest disease prevalence rates among sheep (27.8%) and goats (30.9%) occurred in the spring, and the highest prevalence rates among camels (41.3%) and cattle (80%) were in the summer. Regarding the prevalence of the disease in four slaughterhouses in the Al-Jouf Province, the highest prevalence in sheep, goats, and camels was in the Tabarjal slaughterhouse (1.43%, 0.81%, and 1.08%, respectively), although the Al-Qurayat slaughterhouse had the highest prevalence rate in cattle (1.98%). Complete molecular analysis indicated that cytochrome c oxidase subunit 1 (cox1) sequences from cyst isolates belonged to Echinococcus granulosus (E. granulosus). Moreover, there was high homology (98-100%) with associated Genbank sequences of E. granulosus isolated from sheep in the Kingdom of Saudi Arabia (KSA). Sheep and camels were a significant source of hydatidosis transmission to dogs and helped to maintain disease incidence in the Al-Jouf Province. Thus, significant efforts should focus on preventing cyst transmission from abattoirs and infected stray dogs.

Echinococcus granulosus cyst fluid inhibits the type I interferon response by promoting ROS in macrophages

In cystic echinococcosis (CE), Echinococcus granulosus cystic fluid (EgCF) could impede macrophage-mediated immunity. However, whether EgCF is implicated in the type I interferon response remains to be established. Here, we revealed that EgCF reduced 2′3′-cGAMP-induced IFN-β production in macrophages by inhibiting the cGAS-STING-IRF3 signaling. EgCF also increased the intracellular reactive oxygen species (ROS) levels. Administration of the ROS inhibitor N-acetylcysteine (NAC) restored the cGAS-STING-IRF3 signaling, which, in turn, upregulated IFN-β expression. The findings disclose that EgCF could increase macrophage ROS levels, thereby blocking cGAS-STING-IRF3 signaling and repressing the IFN-I response.

Therapeutic effect of Echinococcus granulosus cyst fluid on bacterial sepsis in mice

BackgroundThe primary pathophysiological process of sepsis is to stimulate a massive release of inflammatory mediators to trigger systemic inflammatory response syndrome (SIRS), the major cause of multi-organ dysfunction and death. Like other helminths, Echinococcus granulosus induces host immunomodulation. We sought to determine whether E. granulosus cyst fluid (EgCF) displays a therapeutic effect on sepsis-induced inflammation and tissue damage in a mouse model.MethodsThe anti-inflammatory effects of EgCF were determined by in vitro culture with bone marrow-derived macrophages (BMDMs) and in vivo treatment of BALB/C mice with cecal ligation and puncture (CLP)-induced sepsis. The macrophage phenotypes were determined by flow cytometry, and the levels of cytokines in cell supernatants or in sera of mice were measured (ELISA). The therapeutic effect of EgCF on sepsis was evaluated by observing the survival rates of mice for 72 h after CLP, and the pathological injury to the liver, kidney, and lung was measured under a microscope. The expression of TLR-2/MyD88 in tissues was measured by western blot to determine whether TLR-2/MyD88 is involved in the sepsis-induced inflammatory signaling pathway.ResultsIn vitro culture with BMDMs showed that EgCF promoted macrophage polarization to M2 type and inhibited lipopolysaccharide (LPS)-induced M1 macrophages. EgCF treatment provided significant therapeutic effects on CLP-induced sepsis in mice, with increased survival rates and alleviation of tissue injury. The EgCF conferred therapeutic efficacy was associated with upregulated anti-inflammatory cytokines (IL-10 and TGF-β) and reduced pro-inflammatory cytokines (TNF-α and INF-γ). Treatment with EgCF induced Arg-1-expressed M2, and inhibited iNOS-expressed M1 macrophages. The expression of TLR-2 and MyD88 in EgCF-treated mice was reduced.ConclusionsThe results demonstrated that EgCF confers a therapeutic effect on sepsis by inhibiting the production of pro-inflammatory cytokines and inducing regulatory cytokines. The anti-inflammatory effect of EgCF is carried out possibly through inducing macrophage polarization from pro-inflammatory M1 to regulatory M2 phenotype to reduce excessive inflammation of sepsis and subsequent multi-organ damage. The role of EgCF in regulating macrophage polarization may be achieved by inhibiting the TLR2/MyD88 signaling pathway.Graphical

Echinococcus granulosus cyst fluid inhibits KDM6B-mediated demethylation of trimethylated histone H3 lysine 27 and interleukin-1β production in macrophages

BackgroundEchinococcus granulosus can manipulate its host's immune response to ensure its own survival. However, the effect of histone modifications on the regulation of the NOD-like receptor protein 3 (NLRP3) inflammasome and downstream interleukin-1β (IL-1β) production in response to the parasite is not fully understood.MethodsWe evaluated IL-1β secretion through enzyme-linked immunosorbent assay and assessed reactive oxygen species levels using the dichlorodihydrofluorescein diacetate probe. Western blotting and quantitative real-time polymerase chain reaction were performed to examine the expression of NLRP3 and IL-1β in mouse peritoneal macrophages and Tohoku Hospital Pediatrics-1 cells, a human macrophage cell line. The presence of trimethylated histone H3 lysine 27 (H3K27me3) modification on NLRP3 and IL-1β promoters was studied by chromatin immunoprecipitation.ResultsTreatment with E. granulosus cyst fluid (EgCF) considerably reduced IL-1β secretion in mouse and human macrophages, although reactive oxygen species production increased. EgCF also suppressed the expression of NLRP3 and IL-1β. Mechanistically, EgCF prompted the enrichment of repressive H3K27me3 modification on the promoters of both NLRP3 and IL-1β in macrophages. Notably, the presence of EgCF led to a significant reduction in the expression of the H3K27me3 demethylase KDM6B.ConclusionsOur study revealed that EgCF inhibits KDM6B expression and H3K27me3 demethylation, resulting in the transcriptional inhibition of NLRP3 and IL-1β. These results provide new insights into the immune evasion mechanisms of E. granulosus.Graphical

Echinococcus granulosus cyst fluid inhibits inflammatory responses through inducing histone demethylase KDM5B in macrophages

BackgroundEchinococcus granulosus cyst fluid (EgCF) weakens macrophage inflammatory responses, thereby enabling the parasite to evade the immune system. However, the role of histone modification in this process remains to be explored.MethodsThe levels of IL-6, TNF-α, IL-10, H3K4me3, and KDM5B were detected using quantitative real-time PCR, ELISA, and Western blotting. The enrichment of H3K4me3 and KDM5B at the promoter of inflammatory factors was detected by chromatin immunoprecipitation.ResultsBased on EgCF-stimulated macrophage models, we found that EgCF significantly inhibited mRNA expression and protein secretion of IL-6 and TNF-α and upregulated mRNA expression of IL-10 under the influence of TLR4. EgCF lowered the level of H3K4me3 and promoted the transcription and protein stability of histone demethylase KDM5B. Chromatin immunoprecipitation analysis revealed that EgCF suppressed the enrichment of H3K4me3 modification at the promoters of TNF-α and IL-6 and downregulated their expression in macrophages. Additionally, the inhibition of KDM5B activity by CPI-455 weakened the anti-inflammatory effect of EgCF.ConclusionsOur findings demonstrate a novel mechanism through which EgCF promotes KDM5B expression and inhibits the enrichment of H3K4me3 at the promoters of inflammatory cytokines to suppress the inflammatory response.Graphical

Phytochemical profile and in vitro protoscolicidal effects of Juniperus phoenicea L., Calotropis procera (Aiton) Dryand, and Artemisia judaica L. against Echinococcus granulosus cysts

Context: Echinococcus granulosus (E. granulosus) is a significant zoonotic agent in veterinary and medicinal fields. Surgery and benzimidazole drugs are used as treatments but have significant drawbacks. Therefore, there is growing interest in ethnomedical approaches to investigating the scolicidal properties of plants traditionally as anthelmintics. Calotropis procera, Juniperus phoenicea, and Artemisia judaica are three plants traditionally used as anthelmintics. Aims: To examine the phytochemical composition and scolicidal capability against E. granulosus cysts of C. procera, J. phoenicea, and A. judaica methanolic extracts. Methods: Fresh E. granulosus cysts were isolated from livestock in southern Jordan and tested to determine the scolicidal potential of C. procera, J. phoenicea, and A. judaica aerial parts methanolic extracts, and their chemical compositions were examined using LC-MS. Results: Although all treatments were protoscolicidal, that of J. phoenicea exhibited higher protoscolices mortality rates at lower concentrations and treatment times than the other extracts. The LC-MS analysis revealed several components with biologically active properties were present in the plant extracts, including terpenes and polyphenolic compounds. Only C. procera contained the steroid uzarin and the flavonoid isoquercitrin. Conclusions: This study demonstrated the potent effects of A. judaica, J. phoenicea, and C. procera methanolic extracts against E. granulosus protoscoleces in vitro, indicating that these plants and/or their phytochemical components may be attractive sources of novel scolicidal medications.

Dogs as a source for the spreading of enteric parasites including zoonotic ones in Giza Province, Egypt

To investigate the impact of domestic and stray dogs on the transmission of zoonotic and other parasites to humans in contact with them, fecal samples were collected from 80 domestic dogs that presented at a clinic with health disturbances and 220 randomly selected stray dogs housed in shelters. The parasitological examination of these samples revealed infection by six zoonotic and four non-zoonotic parasites in varying percentages. The zoonotic parasites included Ancylostoma caninum, Toxocara canis, Dipylidium caninum, Echinococcus granulosus, Cryptosporidium species, and Giardia cysts and trophozoites. The other parasites included Toxascaris leonina, Trichuris vulpis, Taenia species eggs, and Isospora canis oocysts. The infection rate was higher in stray dogs (60%) than in domestic dogs (40%). Infected dogs in both groups were generally unhealthy, with poor body condition recorded in 13.8% of domestic dogs and 63.6% of stray dogs. The infection rate was higher (92%) among shelter workers than among domestic dog owners (66.7%). Giardia assemblages A and D from dogs and assemblage A from humans, as well as two isolates of Cryptosporidium canis (C. canis), one from dogs and the other from humans, were submitted in the GenBank with the accession numbers OQ870443, OQ870444, and OQ919265 for Giardia and OQ917532 & OQ915519 for C. canis of dogs & human, respectively. In conclusion, domestic and stray dogs play an essential role in transmitting zoonotic parasites to humans in contact with them, and regular deworming and strict hygienic measures are recommended to minimize their impact on human health.

PREVALENCE OF Echinococcucus granulosus IN STRAY AND HOME OWNED DOGS IN ADAMAWA STATE, NIGERIA

Echinococcosis is a zoonotic larval infection affecting both domestic animals and human with serious health and economic impacts. A study to determine the prevalence of E. granulosus was carried out on dogs from six Local Government Areas (LGA) of Adamawa State. Faecal samples from 384 dogs were collected and screened for the presence of E. granulosus cyst microspically. A structured questionnaire was used and obtained native and exotic data from dog owners. Out of the 384 dog faecal samples, 171 had E. granulosus cyst with overall prevalence of 44.5% where Guyuk LGA had highest prevalence of 53.1% and Michika LGA had a lowest prevalence of 39.1%). With regards to gender, male and female dogs had a prevalence of 30.5% and 32.4% respectively. In terms of season, highest prevalence of 42.7% and lowest prevalence of 33.3% was recorded during rainy and dried season respectively with a significant association (P&lt; 0.05). Un-restricted dogs recorded a prevalence of 66.1% and 38.1% was recorded in restricted dogs with significant relationship (P&lt; 0.05). Dogs living in suburban areas and kept for security purposes, hunting or guarding have higher chances of infection. Therefore, right and regular treatment of dogs with praziquantel is highly recommended to reduce risk of E. granulosus transmission to humans.

Echinococcus granulosus. Клинический случай

Echinococcosis is a serious problem for modern mankind. This disease has no specific clinical manifestations, but its complications can be fatal. Echinococcus granulosus is endemic for the Astrakhan Region. Every year, the number of cases of human infection with Echinococcus granulosus is increasing on the Astrakhan Region territory. Most often Echinococcus granulosus cysts are found in the liver (85%) and lung (up to 15%). The heart (only 0.4–2%), spleen, kidneys, and brain are damaged much less frequently (1–5% for all locations). The article presents a clinical case of a patient with echinococcosis of the heart and aorta, brain caused by Echinococcus granulosus.

EgCF mediates macrophage polarisation by influencing the glycolytic pathway.

Human cystic echinococcosis (CE) is a zoonotic disorder triggered by the larval stage of Echinococcus granulosus (E. granulosus) and predominantly occurred in the liver and lungs. The M2 macrophage level is considerably elevated among the liver of patients with hepatic CE and performs an integral function in liver fibrosis. However, the mechanism of CE inducing polarisation of macrophage to an M2 phenotype is unknown. In this study, macrophage was treated with E. granulosus cyst fluid (EgCF) to explore the mechanism of macrophage polarisation. Consequently, the expression of the M2 macrophage and production of anti-inflammatory cytokines increased after 48 h treatment by EgCF. In addition, EgCF promoted polarisation of macrophage to an M2 phenotype by inhibiting the expression of transcriptional factor hypoxia-inducible factor 1-alpha (HIF-1α), which increased the expression of glycolysis-associated genes, including hexokinase 2 (HK2) and pyruvate kinase 2 (PKM2). The HIF-1α agonist ML228 also inhibited the induction of macrophage to an M2 phenotype by EgCF in vitro. Our findings indicate that E. granulosus inhibits glycolysis by suppressing the expression of HIF-1α.

Antiparasitic Effects of Asteraceae Species Extracts on Echinococcus granulosus s.s.

Cystic echinococcosis is a zoonotic disease caused by the parasite Echinococcus granulosus sensu lato (s.l.), which is worldwide distributed and causes long-lasting infections in animals and humans. The existing treatment is limited to the use of benzimidazoles, mainly albendazole (ABZ). However, it has unwanted side effects and its efficacy is about 50%. The Asteraceae family includes plants that have therapeutic applications (medicinal species) and has an important role in new drug development. The species belonging to a different genus of this family show a wide range of anti-inflammatory, antimicrobial, antioxidant, hepatoprotective, and antiparasitic activities, among others. The aim of the present study was to evaluate the in vitro efficacy of extracts of four Asteraceae species against protoscoleces of E. granulosus sensu stricto (s.s.). On the other hand, the Stevia aristata extract was assessed on the murine cyst of E. granulosus (s.s.) and the efficacy of S. aristata extract was investigated in a murine model of CE. Stevia satureiifolia, S. aristata, Grindelia pulchella, and G. chiloensis extracts at 100 μg/mL caused a decrease in protoscoleces viability; however, S. aristata extract produced the greatest in vitro protoscolicidal effect. After 20 days of treatment with the highest concentration (100 μg/mL) of S. aristata extract, protoscoleces viability decreased to 0%. The tegumental changes observed by scanning electron microscopy were consistent with the reduction in vitality. The collapse of the germinal layer was registered in 60 ± 5.8% and 83.3 ± 12.0% of cysts treated during 4 days with 50 and 100 μg/ml, respectively. The half maximal effective concentration (EC50) value of the S. aristata extract against E. granulosus (s.s.) cysts was 47.86 μg/mL (96 h). The dosage of infected animals with the 50 mg kg−1 dose of S. aristata extract resulted in a significant reduction in cyst weight in comparison with the control group. In conclusion, S. aristata extract was demonstrated to exert a marked effect, both in vitro and in the murine model.

Improvement of Treatment and Prevention of Liver Echinococcosis

The purpose of the study was to determine optimal pathogenetically justified methods and means to improve the results of diagnosis and treatment of liver echinococcosis. Materials and methods. During treatment, on the 1st, 3rd, 5th, and 10th day of the postoperative period, patients were examined for the functional state of the liver by blood analysis for the amount of proinflammatory cytokines TNF-α, IL-6, IL-4 and IL-10, IL-1. The activity of humoral immunity was assessed based on the level of immunoglobulins IgA, IgG and IgM. Results and discussion. On the 1st day after the operation, the level of IgA increased and amounted to 5.56 ± 0.1 q/L. The improvement in the dynamics of indicators was observed on the 5th day of observations, and this trend persisted up to 10 days after surgical treatment. On day 5, a significant decrease in the concentration of the main proinflammatory cytokines TNF-α, IL-6 and IL-4 was revealed, compared with the indicators before treatment and on the first day after treatment – 12.5 ± 0.6 pg/ml; 11.7 ± 0.3 pg/ml and 4.0 ± 0.2 pg/ml, respectively. This treatment regimen resulted in a marked decrease in TNF-α, IL-6 and IL-10 levels on day 10. The initial level of cytokines and immunoglobulins indicates a pronounced immunosuppressive effect on the body of a parasitic disease. A significant decrease in cytokines IL-1, IL-6 and IL-10 in patients after surgery and a decrease in indicators after maintenance therapy can serve as a potential immunological marker for evaluating its effectiveness. When studying the cytokine profile in the patients we examined, there was a significant increase in the level of TNF-α and IL-6 in blood serum relative to the control values, which is directly dependent on the severity of the pathological process and the functional state of the liver and suppresses the secretion of anti-inflammatory cytokines, which was confirmed by data from foreign authors. As well as the works published earlier, the data of our studies show that Echinococcus granulosus cysts induce a strong antibody response in most patients, causing, at the same time, a reaction of antigen-specific antibodies of various classes with different intensity and specificity against the background of their synthesis and increased secretion. It is necessary to note an increase in quantitative and qualitative indicators of IgG, IgM and IgA levels in the blood serum of patients with liver echinococcosis, with IgG predominance. Conclusion. A comprehensive study of inflammatory mediators and the dynamics of cytokine levels makes it possible to diagnose complications in time and prevent possible relapses of the disease. The data obtained made it possible to justify the further use of imunofan and to develop recommendations for the management of patients with liver echinococcosis in the postoperative period

Breaking-then-curing strategy for efficient cystic echinococcosis therapy

Cystic echinococcosis (CE) is one of the most harmful and life-threatening helminths. As the essential therapeutics, chemotherapy is always difficult to achieve desired anti-echinococcal effect due to the problems that the echinococcus granulosus cyst laminated layer makes the drug difficult to infiltrate and the poor solubility of drugs. In this study, we established a “breaking-then-curing” anti-echinococcal treatment strategy for efficient CE therapy. The photodynamic therapy (PDT) was used as a breaker to produce toxic reactive oxygen species (ROS) and damage the laminated layer of protoscolices (PSCs), leading to enhanced infiltration of albendazole sulfoxide nanoparticles (ABZSO NPs). Then, ABZSO NP was worked as curer for efficient anti-echinococcal treatment. As a result, the breaking-then-curing treatment strategy could generate more intracellular ROS in PSCs induced by plenty of ABZSO NPs, greatly increasing the mortality rate of PSCs in a shorter time than using ABZSO NPs alone, leading to the attenuation of laminated layer and finally disintegrating PSCs. We believe the “breaking-then-curing” strategy will suggest great potential in the treatment of CE and provide a new sight for anti-echinococcal treatment.

The RNA modification in Echinococcus granulosus cysts revealed by mass spectrometry.

RNA modifications, as one of epigenetic mechanisms, are important and conserved mechanisms for maintaining the homeostasis of organisms. Little is known about RNA modifications in Echinococcus granulosus, an obligate parasite that inhabits mammals and gives rise to a huge public health and economic impact. Here, we focused on the RNA modification characteristics of E. granulosus for the first time by using mass spectrometry (UPLC-MS/MS) to qualitatively and quantitatively analyze 47 types of RNA modifications in E. granulosus total RNA. Furthermore, the E. granulosus homologs of writer enzymes preforming RNA modifications were identified, and their gene expression pattern at different developmental stages were analyzed by bioinformatics analysis. Finally, 23 types of RNA modifications were found in E. granulosus cysts total RNA, of which m1A, Ψ and m5C are the most abundant. The homologs of writer enzymes involved in these modifications were identified in the E. granulosus genome, with the dynamic gene expression during the different parasitic developmental stages. This work confirms that E. granulosus retains the conserved RNA modification mechanism during evolution, suggesting the important role of RNA modification in regulating its development and parasitic process. Moreover, the differences of amino acid sequences of RNA modification writer enzymes between parasite and host make it possible to use these enzymes as the candidate drug targets in the follow-up in-depth researches.

Analysis of components of proteins from Echinococcus granulosus cyst fluid

To analyze the components of proteins from Echinococcus granulosus cyst fluid using the shotgun method, and to identify the active components with potential regulatory effects for immune dysregulation diseases. The E. granulosus cyst fluid was collected aseptically from the hepatic cysts of patients with cystic echinococcosis, and characterized by liquid chromatography (LC) tandem mass spectrometry (MS/MS) following digestion with trypsin. The protein data were searched using the software MaxQuant version 1.6.1.0 and the cellular components, molecular functions, and biological processes of the identified proteins were analyzed using the Gene Ontology (GO) method. The E. granulosus cyst fluid separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) had a relative molecular mass of 25 to 70 kDa. LS-MS/MS analysis identified 37 proteins, including 32 known proteins and 5 unknown proteins. At least 4 proteins were preliminarily found to exhibit potential regulatory effects for immune dysregulation diseases, including antigen B, glutathione-S-transferase (GST), thioredoxin peroxidase (TPX) and malate dehydrogenase (MDH). GO enrichment analysis showed that the identified proteins had 149 molecular functions and were involved in 341 biological processes. E. granulosus cyst fluid has a variety of protein components, and four known proteins are preliminarily identified to be associated with immune dysregulation diseases.

Echinococcus granulosus cyst fluid suppresses inflammatory responses by inhibiting TRAF6 signalling in macrophages.

Echinococcus granulosus sensu lato has complex defence mechanisms that protect it from the anti-parasitic immune response for long periods. Echinococcus granulosus cyst fluid (EgCF) is involved in the immune escape. Nevertheless, whether and how EgCF modulates the inflammatory response in macrophages remains poorly understood. Here, real-time polymerase chain reaction and enzyme-linked immunosorbent assay revealed that EgCF could markedly attenuate the lipopolysaccharide (LPS)-induced production of pro-inflammatory factors including tumour necrosis factor-α, interleukin (IL)-12 and IL-6 but increase the expression of IL-10 at mRNA and protein levels in mouse peritoneal macrophages and RAW 264.7 cells. Mechanically, western blotting and immunofluorescence assay showed that EgCF abolished the activation of nuclear factor (NF)-κB p65, p38 mitogen-activated protein kinase (MAPK) and ERK1/2 signalling pathways by LPS stimulation in mouse macrophages. EgCF's anti-inflammatory role was at least partly contributed by promoting proteasomal degradation of the critical adaptor TRAF6. Moreover, the EgCF-promoted anti-inflammatory response and TRAF6 proteasomal degradation were conserved in human THP-1 macrophages. These findings collectively reveal a novel mechanism by which EgCF suppresses inflammatory responses by inhibiting TRAF6 and the downstream activation of NF-κB and MAPK signalling in both human and mouse macrophages, providing new insights into the molecular mechanisms underlying the E. granulosus-induced immune evasion.