Improved experimental model of hepatic cystic hydatid disease resembling natural infection route with stable growing dynamics and immune reaction

AIM To investigate a safer way to set up the disease model of cystic echinococcosis without contamination risk and develop a novel experimental murine model of hepatic cystic echinococcosis. METHODS C57 B/6 mice were injected with human protoscolices of three different concentrations via the portal vein. The mice were followed for 10 mo by ultrasound,gross anatomy,and pathological and immunological examinations. The protoscolex migration in the portal vein,hydatid cyst growth,host immune reaction,and hepatic histopathology were examined periodically.RESULTS The infection rates in the mice in the high,medium,and low concentration groups were 90%,100%,and 63.6%,respectively. The protoscolices migrated in the portal vein with blood flow,settled in the liver,and developed into orthotopic hepatic hydatid cysts,resembling the natural infection route and course.CONCLUSION We have established an improved experimental model of hepatic cystic echinococcosis with low biohazard risk but stable growing dynamics and immune reaction. It is especially useful for new anti-parasite medication trials against hydatid disease.

Efficacy of radiotherapy for the treatment of cystic echinococcosis in naturally infected sheep

Background:Radiotherapy is commonly used to treat cancers.To date,there has been no study focusing on the effects of radiotherapy on hydatid disease in large animals.In this study,we aim to investigate the efficiency and safety of radiotherapy for treating hydatid disease caused by Echinococcus granulosus in naturally infected sheep.Methods:Ultrasound was used to screen naturally infected sheep in an echinococcosis endemic area in Xinjiang,China.A computer tomography(CT)scan confirmed the presence of hydatid cysts.Twenty sheep naturally infected with E.granulosus in the liver and/or lungs were randomly assigned into four groups receiving no irradiation,or X-ray irradiation of low(30 Gy),medium(45 Gy),and high dose(60 Gy),respectively.After three months of radiotherapy,a CT scan was performed to measure the changes in the cysts.The hepatic parasite cysts and host tissues were collected for histology and gene expression analysis.Results:In the animals subject to irradiation,no significant differences were observed in their appetite,daily activities,and weight before and after radiotherapy.Severe calcification was noticed in the cysts subject to a high dose of radiation compared with the groups subject to low and medium doses.Hematoxylin and eosin staining showed that irradiation contributed to the damage of the cyst structure and nucleus in the germinal layers.Quantitative PCR demonstrated that expression of TPX and HSP70 significantly decreased in a dose-dependent manner(P<0.05).The expression of the EPC1 decreased in the medium-and high-dose groups compared with the low-dose group(P<0.05).Meanwhile,the expression of radiation-related apoptosis genes caspase-3 and Gadd45 decreased with an increase in the irradiation dose.Conclusions:Radiotherapy is an option with satisfactory efficiency and safety for treating cystic echinococcosis in sheep with partial response or stable disease at month 3.In future,inhibition of cystic activity using radiotherapy may serve as a new regimen for treating hydatid disease.

Suppression of acute rejective response following orthotopic liver transplantation in experimental rats infected with Echinococcus multilocularis

Background Hepatic alveolar echinococcosis （AE） is a parasitic disease in humans and caused by the Echinococcus multilocularis （Em）. Orthotopic liver transplantation （OLT） may be the only effective treatment for end-stage hepatic AE. However, in some AE patients, extrahepatic Em can not be completely eliminated after OLT. We aimed to study whether the immunological changes caused by Em evasion may influence the rejective response. Methods Rat modles of AE were established by injecting the Em suspension into abdomen of Brown Norway （BN） rats. Three months later, in the experimental group, the liver was transplanted from Lewis （LEW） rats to Em-infected BN rats. In the control group, transplantation was from LEW rats to healthy BN rats. Liver tissue and peripheral blood （PB） samples were collected on days 1, 3, 5, and 7 after OLT. Liver tissue was analyzed after hematoxylin and eosin （H＆E） staining; numbers of CD4, CD8, and CD28 on peripheral blood cells were detected by flow cytometry; and expression of the chemokine fractalkine （Fkn） was detected by reverse transcription PCR （RT-PCR）. Interleukin-10 （IL-10） was measured in the serum by enzyme-linked immunosorbent assay （ELISA）. In every group, eight BN rats were retained for observing survival time. Results The survival times of recipients in the experimental group were prolonged compared with those in the control group. The rejective response occurred later and was milder in the experimental group, percentage of CD4, CD8, CD28 T-cells and Fkn mRNA expression were lower in the experimental group. While the serum IL-10 levels were higher in the experimental group than those in the control group. Conclusions Acute rejective response after OLT was attenuated in the rats with Em infection, and the recipients＂ survival time was prolonged. Em may play a role in this process by elevating IL-10 secretion, decreasing the effector T cells, inhibiting the expression of Fkn, which lead to reduce the inflammatory cells infiltration into the liver.

Molecular characterization of a signal-regulated kinase homolog from Echinococcus granulosus

Background Cystic echinococcosis due to Echinococcus granulosus （E. granulosus） is one of the most important chronic helminthic diseases, especially in sheep/cattle-raising regions. The larval stage of the parasite forms a cyst that grows in the liver, lung, or other organs ofthe host. To ensure a long life in the host tissues, the parasite establishes complex inter-cellular communication systems between its host to allow its differentiation toward each larval stage. Recent studies have reported that this communication is associated with the extracellular signal-regulated kinase （ERK） mitogen-activated protein kinase cascade in helminth parasites, and in particular that these protein kinases might serve as effective targets for a novel chemotherapy for cystic echinococcosis. The aim of the present study investigated the biological function of a novel ERK ortholog from E. granulosus, EgERK. Methods DNA encoding EgERK was isolated from protoscolices of E. granulosus and analyzed using the LA Taq polymerase chain reaction （PCR） approach and bioinformatics. Reverse transcription PCR （RT-PCR） was used to determine the transcription level of the gene at two different larval tissues. Western blotting was used to detect levels of EgERK protein. The expression profile of EgERK in protoscolices was examined by immunofluorescence. Results We cloned the entire Egerk genomic locus from E. granulosus. In addition, two alternatively spliced transcripts of Egerk, Egerk-A, and Egerk-B were identified. Egerk-A was found to constitutively expressed at the transcriptional and protein levels in two different larval tissues （cyst membranes and protoscol（ces）. Egerk-A was expressed in the tegumental structures, hooklets, and suckers and in the tissue surrounding the rostellum of E. granulosus protoscolices. Conclusions We have cloned the genomic DNA of a novel ERK ortholog from E. granulosus, EgERK （GenBank ID HQ585923）, and found that it is constitutively expressed in cyst membrane and protoscolex. These findings will be useful in further study of the biological functions of the gene in the growth and development of Echinococcus and will contribute to research on novel anti-echinococcosis drug targets.

Activation of Anti-tumor Immune Response by Ablation of HCC with Nanosecond Pulsed Electric Field

Locoregional therapy is playing an increasingly important role in the non-surgical management of hepatocellular carcinoma(HCC).The novel technique of non-thermal electric ablation by nanosecond pulsed electric field has been recognized as a potential locoregional methodology for indicated HCC.This manuscript explores the most recent studies to indicate its unique anti-tumor immune response.The possible immune mechanism,termed as nano-pulse stimulation,was also analyzed.

Improved recognition of Echinococcus granulosus protoscoleces using visual saliency and scale-invariant features

Echinococcosis—a parasitic disease caused by Echinococcus granulosus or Echinococcus multilocularis larvae—occurs in many regions in the world. This disease can pose a serious threat to public health and thus requires a convenient and cost-effective method for early detection. So, we developed a novel method based on visual saliency and scale-invariant features that detects the tapeworm parasites. This method improves upon existing bottom-up computational saliency models by introducing a visual attention mechanism. The results indicated that the proposed method offers a higher level of both accuracy and computational efficiency when detecting Echinococcus granulosus protoscoleces, which in turn could improve early detection of echinococcosis.

Study on the echinococcosis blood serum detection based on Raman spectroscopy combined with neural network

A Raman spectroscopy method combined with neural network is used for the invasive and rapid detection of echinococcosis. The Raman spectroscopy measurements are performed on two groups of blood serum samples,which are from 28 echinococcosis patients and 38 healthy persons,respectively. The normalized Raman reflection spectra show that the reflectivity of the echinococcosis blood serum is higher than that of the normal human blood serum in the wavelength ranges of 101—175 nm and 1 801—2 701 nm. Then the principal component analysis(PCA) and back propagation neural network(BPNN) model are used to obtain the diagnosis results. The diagnosis rates for healthy persons and echinococcosis persons are 93.333 3% and 90.909 1%,respectively,so the average final diagnosis rate is 92.121 2%. The results demonstrate that the Raman spectroscopy analysis of blood serum combined with PCA-BPNN has considerable potential for the non-invasive and rapid detection of echinococcosis.

A fluorescent porous silicon-based biosensor for small molecule detection

Fluorescent porous silicon was prepared as a stable biosensor chip substrate. The aminopropyltriethoxysilane(APTES) molecules are attached in the pores of the porous silicon with a crosslink method, and when the molecules are added into the chip, the fluorescence intensity is reduced according to the concentration of the APTES. Controlled experiments are also presented with the small molecule that cannot be covalently coupled, and the results show that this kind of sensor chip has better specificity. Compared with other conventional methods, this method is simple, quick and label-free.