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The cestode Echinococcus multilocularis larva infection causes lethal zoonotic alveolar echinococcosis (AE), a disease posing a great threat to the public health worldwide. This persistent hepatic tumor-like disease in AE patients has been largely attributed to aberrant T cell responses, of which Th1 responses are impeded, whilst Th2 and regulatory T cell responses are elevated, creating an immune tolerogenic microenvironment in the liver. However, the immune tolerance mechanisms are not fully understood. Dendritic cells (DCs) are key cellular components in facilitating immune tolerance in chronic diseases, including AE. Here, we demonstrate that indoleamine 2,3-dioxygenase 1-deficient (IDO1-/-) mice display less severe AE as compared to wild-type (WT) mice during the infection. Mechanistically, IDO1 prevents optimal T cells responses by programming DCs into a tolerogenic state. Specifically, IDO1 prevents the maturation and migration potential of DCs, as shown by the significantly enhanced expression of the antigen-presenting molecule (MHC II), costimulatory molecules (CD80 and CD86), and chemokine receptors (CXCR4 and CCR7) in infected IDO1-/- mice as compared to infected wild-type mice. More importantly, the tolerogenic phenotype of DCs is partly reverted in IDO1-/- mice, as indicated by enhanced activation, proliferation, and differentiation of both CD4+ and CD8+ - T cells upon infection with Echinococcus multilocularis, in comparison with WT mice. Interestingly, in absence of IDO1, CD4+ T cells are prone to differentiate to effector memory cells (CD44+CD62L-); in contrast, CD8+ T cells are highly biased to the central memory phenotype (CD44+CD62L+). Overall, these data are the first to demonstrate the essential role of IDO1 signaling in inducing immunosuppression in mice infected with Echinococcus multilocularis.

BackgroundAlveolar echinococcosis (AE) is a chronic zoonosis caused by the larval form ofEchinococcus multilocularis(E. multilocularis). Current chemotherapy against AE has relied on albendazole and mebendazole, which only exhibit parasitostatic and not parasiticidal efficacy. Therefore, novel compounds for the treatment of this disease are needed.MethodsPhosphoglucose isomerase (PGI) assays were used for compound screening of seven neonicotinoids. The anti-parasitic effects of thiacloprid were then evaluated onE. multilocularismetacestode vesicles, germinal cells and protoscoleces in vitro. Human foreskin fibroblasts (HFF) and Reuber rat hepatoma (RH) cells were used to assess cytotoxicity. Glucose consumption inE. multilocularisprotoscoleces and germinal cells was assessed by measuring uptake of 2-deoxyglucose (2-DG). Molecular docking was used to evaluate the potential binding sites of thiacloprid to acetylcholine receptors. In vivo efficacy of thiacloprid was evaluated in mice by secondary infection withE. multilocularis. In addition, ELISA and flow cytometry were used to evaluate the effects of cytokines and T lymphocyte subsets after thiacloprid treatment. Furthermore, collagen deposition and degradation in the host lesion microenvironment were evaluated.ResultsWe found that thiacloprid is the most promising compound, with an IC50of 4.54 ± 1.10 μM and 2.89 ± 0.34 μM, respectively, against in vitro-culturedE. multilocularismetacestodes and germinal cells. Thiacloprid was less toxic for HFF and RH mammalian cell lines than for metacestodes. In addition, thiacloprid inhibited the acetylcholinesterase activity in protoscoleces, metacestodes and germinal cells. Thiacloprid inhibited glucose consumption by protoscoleces and germinal cells. Subsequently, transmission electron microscopy revealed that treatment with thiacloprid damaged the germinal layer. In vivo, metacestode weight was significantly reduced following oral administration of thiacloprid at 15 and 30 mg/kg. The level of CD4+T lymphocytes in metacestodes and spleen increased after thiacloprid treatment. Anti-echinococcosis-related cytokines (IL-2, IL-4, IL-10) were significantly increased. Furthermore, thiacloprid inhibited the expression of matrix metalloproteinases (MMPs 1, 3, 9, 13) and promoted collagen deposition in the host lesion microenvironment.ConclusionsThe results demonstrated that thiacloprid had parasiticidal activity againstE. multilocularisin vitro and in vivo, and could be used as a novel lead compound for the treatment of AE.Graphical abstract

Additional file 1: Fig. S1. Survival analysis of protoscoleces in metacestodes after crocin treatment. After 6 weeks of treatment, metacestodes were isolated from the abdominal cavity of mice. The metacestode of each mouse was cut into pieces by the same person, and the contents were filtered through gauze into a 50 ml centrifuge tube. The contents were centrifuged at 5000 rpm for 5 min, and the precipitation was stained with 0.1% eosin. The number of live and dead protoscoleces was counted under a microscope. Scale bar = 200 μm. A Protoscolex morphology isolated from mouse metacestodes. The blue arrows show the calcareous body, the red arrows show the live protoscoleces, and the black arrows show the dead protoscoleces. B Total number of protoscoleces. C The viability of the protoscoleces was assessed by eosin staining.

In this paper, we study the existence of multiple positive solutions for a class of fractional Schr��dinger-Poisson systems involving sign-changing potential and critical nonlinearities on an unbounded domain. With the help of Nehari manifold and Ljusternik-Schnirelmann category, we investigate how the coefficient $g(x)$ of the critical nonlinearity affects the number of positive solutions. Moreover, we present a novel relationship between the number of positive solutions and the category of the global maximum set of $g(x)$., 44pages

Supplemental material, sj-pdf-2-imr-10.1177_03000605211014395 for Pancreatic pseudocyst, pancreatitis, and incomplete pancreas divisum in a child treated with endotherapy: a case report by Jingxin Yan, Zheheng Zhang, Zhixin Wang, Wenhao Yu, Xiaolei Xu, Yaxuan Wang and Haining Fan in Journal of International Medical Research

Supplemental material, sj-pdf-1-imr-10.1177_03000605211014395 for Pancreatic pseudocyst, pancreatitis, and incomplete pancreas divisum in a child treated with endotherapy: a case report by Jingxin Yan, Zheheng Zhang, Zhixin Wang, Wenhao Yu, Xiaolei Xu, Yaxuan Wang and Haining Fan in Journal of International Medical Research

Additional file 1: Figure S1. Chemical structure and functional groups of neonicotinoids. a Chemical structure of neonicotinoids. Chemical structure obtained from PubChem database. b Functional group information of neonicotinoids.

Additional file 8: Figure S5. The cell viability of protoscoleces and germinal cells treated with thiacloprid for 1 h. a Survival of protoscoleces after treatment with thiacloprid. To evaluate the survival of protoscoleces, 0.1% eosin staining exclusion method was used. b Mitochondrial probe labeling of living cells.

Additional file 2: Fig. S2. SEM observation of isolated protoscoleces from metacestode in mouse. In the control group and ABZ group, intact protoscoleces were observed, and the protoscoleces were invagination type. After treatment with crocin, the body wall of the protoscoleces appeared wrinkled. Representative images are displayed separately. Scale bar = 50 μm.

Additional file 3: Figure S3. Isolation of metacestodes from Mongolian gerbil. The black arrows show the protoscoleces.

Additional file 12: Figure S8. Immune cell infiltration in host tissue surrounding metacestode. After thiacloprid treatment, infiltration of neutrophils decreased (a) and infiltration of plasma cells, lymphocytes and macrophages increased in host tissue surrounding metacestode (b). The red arrow shows the neutrophils, the black arrow shows the lymphocytes, the green arrow shows the eosinophils, the blue arrow shows the plasma cells, and the yellow arrow shows the macrophages.

Additional file 5: Table S2. Antibody information.

Additional file 6: Figure S4. Metacestode vesicle (a) and germinal cells (b) cultured in vitro.

Supplemental material, sj-pdf-1-imr-10.1177_03000605211014395 for Pancreatic pseudocyst, pancreatitis, and incomplete pancreas divisum in a child treated with endotherapy: a case report by Jingxin Yan, Zheheng Zhang, Zhixin Wang, Wenhao Yu, Xiaolei Xu, Yaxuan Wang and Haining Fan in Journal of International Medical Research

Additional file 7: Table S3. Echinococcus multilocularis cholinesterase activity.

Supplemental material, sj-pdf-2-imr-10.1177_03000605211014395 for Pancreatic pseudocyst, pancreatitis, and incomplete pancreas divisum in a child treated with endotherapy: a case report by Jingxin Yan, Zheheng Zhang, Zhixin Wang, Wenhao Yu, Xiaolei Xu, Yaxuan Wang and Haining Fan in Journal of International Medical Research

Additional file 11: Table S4. Blood cell analysis of mice infected with E. multilocularis treated with thiacloprid (n = 6).

Additional file 1. Definitions and references of some abbreviations in the article.

Additional file 10: Figure S7. The center of the metacestodes showed caseous necrosis. HE staining showed caseous necrosis in the centre of the metacestode (scale bar = 400 μm). There is no structural granular and no residual shadow of the original tissue structure.

Additional file 4: Table S1. Specific primer sequences.

Hydatidosis is a zoonotic parasitic disease caused by Echinococcus, which is highly prevalent in pastoral areas. In China, this disease is mainly caused by Echinococcus granulosus and Echinococcus multilocularis. Cystic echinococcosis, which is one of the most common types of echinococcosis, is described as swelling and growth of cystic lesions. Alveolar echinococcosis, which is less common, is invasive. Cases of mixed echinococcosis infection accompanied by extrahepatic organ metastasis are extremely rare. Treatment of these cases is complicated and the prognosis is extremely poor. We report a case of hepatic echinococcosis in a 40-year-old Tibetan man who was treated with the hepatic right tricuspidectomy + left hepatic duct jejunostomy (Roux-en-Y) surgical procedure. This procedure provides a reference for treatment of similar cases of echinococcosis. For patients with multiorgan metastasis, chemotherapy is the first choice. This should be followed by possible surgical treatment for life-threatening lesions of alveolar echinococcosis and subsequent chemotherapy. Individualized treatment accompanied by multidisciplinary treatment and damage control surgery could optimally benefit patients with advanced hepatic echinococcosis.

A new method is presented for blind parameter estimation of frequency hopping signals. According to the inner structure of frequency hopping signals, the three-parameter frequency-time atom is obtained from the unit windowed sine function by successive applications of scaling, time-shift and frequency-shift operators. On the base of these, a new matching pursuit based parameter estimation method of frequency hopping signal is developed. Using this method, the parameter of frequency hopping signals i.e. as hop duration, hop timing and hopping frequencies can be estimated without any priori knowledge. Simulation results demonstrate that the method is effective and efficient.

It is shown that if 2 is a primitive root mod prime n+1 and GF(2/sup n/) is generated by an irreducible all one polynomial (AOP) then a simple multiplication algorithm exists over the field GF(2/sup n/).