Human cystic echinococcosis in Heilongjiang Province, China: a retrospective study.

BACKGROUND: Cystic echinococcosis (CE) is one of emerging zoonotic parasitic diseases throughout the world, having significant medical and economic importance in developing countries. The western and northwestern China is considered as CE endemic areas. In northeastern China's Heilongjiang Province, the increasing number of sporadic human CE cases has attracted more and more attention. The aims of the present study were to understand the clinical characteristics of human CE in the investigated area and to compare the coincidence rates of CT, ultrasound and serological test against the histopathology results among CE patients. METHODS: Hospital data of 183 human CE cases in the period from January 2004 to July 2013 were collected from the two largest hospitals in Heilongjiang Province. Clinical data were analyzed, including age, gender, occupation and living residence of CE patients and localization, size and number of CE cysts as well as the diagnosis methods of CE before operation. RESULTS: The results revealed that the incidence of CE reached a peak in the age group of 41-50 years. Among the 183 CE patients, the females were observed to have a higher percentage of CE patients (60.66%, 111/183) than males (39.34%, 72/183). The majority of CE patients were farmers, followed by workers, employees, public servants, students and so on. CE cysts were most commonly found in the livers, with a 30 cm cyst in diameter being detected. CT showed the highest coincidence rate (96.64%) for hepatic CE among the three common diagnosis methods (CT, ultrasound imagine and serological test) compared against the histopathology results. CONCLUSIONS: This is the first retrospective analysis of human CE cases in Heilongjiang Province in recent ten years. Clinical characteristics of human CE were described here. CT appeared to be the most effective diagnosis method for hepatic CE.

Does hydatid disease have protective effects against lung cancer?

We hypothesized that solid tumors rarely occur in patients with hydatid disease. We obtained the serum of 14 patients diagnosed with hydatid disease, the serum of 10 patients who did not have a history of hydatid disease, and the hydatid cyst fluid from six patients. These sera and fluid samples were added at different concentrations to NCI-H209/An1 human lung small cell carcinoma cells and L929 mouse fibroblasts as a control group. Sera of patients with hydatid diseases had cytotoxic effects on NCI-H209/An1 cells, but they did not have cytotoxic effects on fibroblast cells. Sera from healthy subjects did not have a cytotoxic effect on the tumor cell line or control fibroblasts. Cyst fluid, also, did not have toxic effects on the NCI-H209/An1 cell line, but was toxic to fibroblasts up to a 1:32 dilution. Sera from patients with hydatid disease had cytotoxic effects on human small cell lung cancer cells in vitro.

Proteomics investigation of human sera for determination of postoperative indicators of pulmonary cystic echinococcosis.

BACKGROUND: Cystic echinococcosis (CE)/hydatidosis is an important zoonotic parasitic disease caused by the larval stage of Echinococcus granulosus. The disease is a major health problem all over the world. Finding specific and sensitive biomarkers for follow-up of CE in patients after surgery is essential. Using proteomics methods, the present study aimed to evaluate post-surgical treatment by finding probable biomarker/s in the serum of human lungs CE. METHODS: A total of 24 human sera were tested. These sera included eight confirmed lung/s CE patients sera before surgery (BS), eight sera 12 months post-surgery (12MPS) as well as eight control sera from healthy people. Proteomics methods including 2DE and LC-MS/MS were performed on the specimens followed by bioinformatics analysis. Differentially expressed proteins (DEP) were detected and, separately integrated with protein-protein interaction (PPI) data to construct the PPI network. RESULTS: A total of 171 protein spots were detected in three groups including BS, 12MPS, and control groups; of which a total of 106 DEP have been expressed based on fold changes > = 2 and p-value < 0.05. More analysis was performed and a total of 10 protein spots were selected for identification by mass spectrometry showing the following proteins: APOA1, BGN, SPP2, EAF1, ACOXL, MRPL55, MCTP2, SEPTIN1, B4GALNT1, and ZNF843. Based on centrality parameters of the PPI network (degree and betweenness) five Hub-bottlenecks proteins with significant centrality values were found including APOA1, BGN, SPP2, EAF1, and ACOXL. CONCLUSION: This study showed five proteins as hub-bottleneck proteins; of which APOA1 was more prominent. It can be concluded that a change in expression of this protein in patients' sera could be used as an indicator tool for the achievement of lungs CE surgical therapy.

Evaluation of pre- and post-surgical oxidative stress parameters in patients with pulmonary cystic echinococcus.

BACKGROUND: Hydatid cyst secretes numerous immunomodulatory molecules to the host, and the host reacts these molecules by activating immune response. It is also known that, immune cells generally produce more oxidative products. AIM: In this study, we aimed to evaluate the antioxidant enzyme using catalase (CAT) and oxidant enzyme using lipid hydroperoxide (LOOH) levels in the serum samples before and after the surgical interventions in patients with pulmonary cystic echinococcus (CE). PATIENTS AND METHODS: Forty patients with pulmonary CE who underwent surgery and 40 healthy individuals were enrolled to the study. Patients were divided in two groups; group 1 (n=40) consisted of patients with pulmonary CE, and group 2 (n=40) consisted of healthy subjects. Before and after surgical interventions serum CAT and LOOH levels were measured. RESULTS: Compared to group 2, group 1 had significantly lower CAT and higher LOOH levels before surgery (both p < 0.001). In group 1; CAT levels were significantly increased and LOOH levels significantly increased after the surgical intervention (both p < 0.001). CONCLUSIONS: The present study showed that oxidative stress increased in patients with pulmonary CE, may be consequence of immune response of the host, and these levels decreased after the surgical intervention.

Comparative analysis of the diagnostic performance of crude sheep hydatid cyst fluid, purified antigen B and its subunit (12 Kda), assessed by ELISA, in the diagnosis of human cystic echinococcosis.

The diagnosis of patients with cystic echinococcosis (CE) by means of serology has a limited support in clinical practice due to cross-reactivity with other helminthes leading to overestimation of the parasite's true prevalence. A wealth of reports on the diagnostic performance of antigen B (AgB) has been produced. This study was designed to comparatively assess the diagnostic efficacy of crude sheep hydatid cyst fluid (HCF), AgB and its subunit (12 KDa) to detect IgG or IgG4 antibodies in CE patients' sera using enzyme-linked immunosorbent assay (ELISA).The best diagnostic performance was obtained with anti-HCF IgG ELISA which gave 92.4% sensitivity and 92.6% specificity. Despite the low sensitivity of anti AgB IgG ELISA (84%), it gave the best specificity (94.4%) with less cross-reaction with sera of subjects infected with other parasites. In conclusion, it is recommended to use anti-HCF IgG ELISA for initial screening in large seroprevalence studies. Further analysis of positive serum samples with anti AgB IgG ELISA would allow the confirmation of true positives. Specific IgG4 ELISA may represent a complementary assay, useful as secondary confirmatory tests for patients with suspected CE and negative for total IgG ELISA.

[Contribution of Western blotting to the diagnosis of hydatidosis]. / Apport de la technique Western blot dans le diagnostic de l'hydatidose.

The aim of this study is to evaluate the contribution of the immunoWesternblot for the diagnosis and the post surgical follow-up of the hydatidosis. 71 sera from patients with hydatidosis confirmed by surgery were studied. All had a negative hydatic serology by screening tests (enzyme-linked immunosorbent assay, hemagglutination, electrosyneresis). 12 patients with sera in pre and post operative were monitored for 2 years. The Echinococcus Western blot IgG permitted to rectify the diagnosis of hydatidosis in 67.6 %. The rate of positivity was 100 % for the multivesicular liver cysts, 60 % for the young cysts and 50 % for the calcified cysts. Western blot permitted to rectify the diagnosis of lung cysts in 62.5 % of cases and in 50 % of cranial-spinal localizations. Analysis of Western Blot evolution in the 12 patients followed in pre and post-surgical revealed the disappearance of the bands 16, 18 and 26-28kDa in 8 month in the 8 patients with complete exeresis. This study proved the value added of Western blot compared to the other traditional techniques for the immunodiagnostic and the post-surgical monitoring of hydatidosis.

Concurrent pulmonary sequestration and pulmonary hydatid cyst in a child: A case report.

Pulmonary sequestration (PS) happens when an area of the lung receives its arterial blood supply from systemic circulation, resulting in a non-functional lesion (Intralobar or extralobar). Hydatid cyst results from infection of the tapeworm Echinococcus. We report a case of hydatid cyst existed concurrently with intralobar PS. A 12-year-old girl presented with recurrent hemoptysis of 2 years duration. Serology for Echinococcus granulosus was positive. CT chest suggested intralobar PS in the right middle lobe, which was surgically removed. In conclusion, recurrent localized pulmonary infections should raise the suspicion of intralobar PS which may rarely coexist with hydatid cyst.

Comparison of serologic tests for the diagnosis and follow-up of alveolar hydatid disease.

Alveolar hydatid disease is a serious and often fatal condition caused by infection with the metacestode form of Echinococcus multilocularis. Sera of 21 patients with histologically confirmed disease were tested by an enzyme-linked immunosorbent assay (ELISA) using a semi-purified E. multilocularis antigen fraction (Em2) and by indirect hemagglutination (IHA) and double diffusion (DD5) tests using antigens prepared from E. granulosus cyst fluid. At diagnosis, sera from all 21 patients were positive by Em2 ELISA, 18 (86%) by IHA, and 5 (24%) by DD5. Em2 ELISA detected an antibody response earlier than IHA in 4 of 9 patients from whom sera were available before diagnosis. Following complete surgical resection, Em2 ELISA converted from positive to negative in serum of 2 of 3 patients, while IHA results did not change. Following incomplete resection, 14 of 15 patients tested remained positive by Em2 ELISA, while 12 remained positive by IHA. Of sera from 361 healthy persons from regions free of E. multilocularis, none were positive by Em2 ELISA, while 8% were positive by IHA. Of sera from 59 patients with non-echinococcal parasitic infections, none were positive by Em2 ELISA, while 31% were positive by IHA. Thus, in comparison with tests using E. granulosus antigens, Em2 ELISA appears to be more sensitive and specific for diagnosing AHD, useful on follow-up of resected patients, and positive earlier in the course of disease.

Western blot applied to the diagnosis and post-treatment monitoring of human hydatidosis.

The serologic diagnosis of hydatidosis (caused by Echinococcus granulosus) can be made by different techniques, although the lack of standardization of the antigens affects the sensitivity, specificity and concordance among the different tests. We have applied the Western-Blot (WB) technique, associated with a purified antigen from sheep hydatid fluid, at 60 samples of serum from 14 patients suffering echinococcosis in different bodily locations, monitored for 3 years. The WB test enabled the detection of antibodies in the pre-surgical samples for proteins of 12-14, 16, 20, 24-26, 34, 39 and 42 kDa in molecular weight in 15-96% of the patients. The combination involving 2 of the 3 proteins of 20, 39 and 42 kDa has made it possible to diagnose 100% of the cases. The antibodies specific to proteins 39 and 42 kDa disappeared in less than one year in the patients cured after surgery, while in patients with persistent or recurrent parasitism the bands present before surgery persisted or other new ones appeared. The WB with purified antigens proved to be highly useful in the diagnosis and post-surgical monitoring of hydatidosis patients. The antigen used is proposed as a standard antigen for the diagnosis and follow-up of pre- and postsurgical hydatidosis.

Interleukin (IL)-4, IL-10 and IL-12 profile in serum of patients with alveolar echinococcosis.

Alveolar echinococcosis (AE), caused by Echinococcus multilocularis (E.m.), provokes a characteristic immune response. Based mainly on in vitro studies, Th2 dominated immunity is associated with increased susceptibility to disease, while Th1 cell activation is assumed to induce protective immunity. We investigated serum levels of interleukin (IL)4, IL-10, and IL-12 in 40 AE patients and 20 controls to assess Th1/Th2 cell activation in vivo. Significantly higher levels of IL-10 were found in AE patients (P = 0.003) than in controls, with a tendency to higher concentrations in progressive cases. In contrast, IL-4 was only measurable in a minority of patients and controls. IL-12 levels (measured with an ELISA that detects both the p35/p40 heterodimer and free p40) were comparable between AE patients and controls and showed a similar distribution pattern to IL-10 with regard to disease progression. By using an IL-12-ELISA specific for the heterodimer, only minute amounts of IL-12 were detectable in merely a minority of samples. In conclusion, our data are suggestive of Th2 dominated immune response in AE in vivo.

Increased basal production of interleukin-10 by peripheral blood mononuclear cells in human alveolar echinococcosis.

The secretion of IL-10 by peripheral blood mononuclear cells (PBMC) and the expression of IL-10 mRNA in fractionated CD4+ and CD8+ lymphocyte subsets and non-B-non-T cells, with and without stimulation by the mitogen phytohemagglutinin-C (PHA-C) and specific Echinococcus multilocularis (E. multilocularis) antigens, were assessed in 7 patients with alveolar echinococcosis (AE) and 6 healthy subjects. Results of studies on IL-10 were compared to those on IFN-gamma, IL-4 and IL-5 in the same patients and control subjects. IL-10 production was significantly higher in patient PBMC-culture supernatants than in the control group supernatants, both at the basal level and after mitogen or specific E. multilocularis antigen stimulation. Both CD4+ and CD8+ lymphocyte populations and non-B-non-T cells of AE patients and controls expressed IL-10 mRNA. Semi-quantification of IL-10 mRNA revealed a significantly higher transcript level in unstimulated-CD8+ T cells from AE patients in comparison with CD8+ T cells of healthy donors. PBMC from patients produced very low levels of IL-4 but the production of IFN-gamma was not significantly depressed compared to the controls. PBMC, isolated from 4 AE patients and 4 control subjects stimulated with specific E. multilocularis antigens, secreted IL-5; IL-5 mRNA was only detected in the CD4+ lymphocyte subset. The secretion of IL-5 and the expression of IL-5 mRNA in healthy subjects could be due to the presence of non-specific mitogenic parasitic factors. This non-specific mitogenic activity of the parasite, besides inducing a high secretion of IL-10 in patients with evolutive AE, may contribute to the lack of host control of parasite growth and to the persistence of granulomatous lesions, due to the inhibition of an efficient Th1 immune response.

[Laboratory evaluation of commercial immunoblot assay kit for serodiagnosis of Echinococcus infections using sera from patients with alveolar hydatidosis in Hokkaido].

Using serum specimens from patients with alveolar hydatidosis (AH) in Hokkaido, we assessed the usefulness of "Echinococcus Western Blot IgG" (the French immunoblot assay, FIA), which has recently been launched from Ldbio Diagnostics (Lyon, France) as new commercial immunoblot assay kit of immunodiagnosis of Echinococcus infections. Eighty serum specimens were used for the present study: 64 preoperative sera and nine postoperative sera, which were taken from AH patients in Hokkaido, and seven sera from persons who were ELISA (enzyme-linked immunosorbent assay)--positive in mass screening which was conducted for checking on Echinococcus infections in Hokkaido since 1982. When the 64 preoperative sera were examined by the Western blotting method (the Hokkaido method of Western blotting, HWB) which had been carried out at Hokkaido Institute of Public Health between 1987 and 1993, it was found that 53 cases were positive and six cases were quasi-positive, i.e. the rate of the positive cases including quasi-positive cases was 92.2%. From immunostaining patterns, HWB-positive sera could be grouped in two types: the complete type, which showed a pattern of multiple bands containing the 55 and 66 kDa bands, and the incomplete type, which showed patterns of only few bands containing the AH-specific polysaccharide antigen named C antigen. Forty-three of the 53 HWB-positive sera were of the complete type and the residue was of the incomplete type. On the other hand, when the 64 preoperative sera were examined by FIA, 60 sera (93.8%) were judged to be positive and the others as negative sera. On the basis of the interpretation of immunostaining patterns described in the instruction manual, 47 (78.3%) of the 60 positive sera were regarded as pattern P3, five (8.3%) as pattern P4, and eight (13.3%) as pattern P5. All of the complete-type sera were regarded as P3, indicating high antibody titers. Contrarily, most of the incomplete-type or quasi-positive sera resulted in other patterns such as P4 and P5, indicating low antibody titers. Of 5 HWB-negative sera, two were FIA-positive (which showed P3 and P5 patterns respectively), however their immunoreactions were significantly low. Therefore, apart from interpretation of pathological conditions of cases with exceedingly low antibody titers, FIA may be able to give a serologically clear interpretation to HWB-quasi-positive cases, indicating that it is a highly sensitive and useful method for immunodiagnosis of Echinococcus infections.

[The interferon status in helminthiases. 1. Human unilocular hydatid disease]. / Interferonovyi status pri gel'mintozakh. Soobshchenie 1. Ekhinokokkoz odnokamernyi cheloveka.

In 51 patients with Echinococcus granulosus infection, blood IF alpha and IF gamma levels were measured using vesicular stomatitis virus culture. In patients with progressing disease, the IF alpha and IF gamma levels were significantly lower than in the benign form of the disease--144 +/- 28.8 and 51.6 +/- 10.1 U/ml, respectively (in healthy controls the levels were 1024 +/- 28.8 and 187.7 +/- 16.9 U/ml). The lowest levels of IF alpha and IF gamma were found in patients with primary localization of cysts in the lungs and multiorgan damages affecting the lungs--160 +/- 38.5 and 44.4 +/- 13.5 U/ml; 146.7 +/- 80.1 and 89.5 +/- 19.2 U/ml, respectively. After 5-6 thirty-day courses of mebendazole therapy (50 mg/kg body weight) at 30-day intervals, the average levels of IF alpha and IF gamma were 4 and 5 times higher. Measurement of IF levels in echinococcosis may be of prognostic value for the estimation of the efficacy of surgical and/or specific treatment.

[The interferon status in helminthiases. 2. Alveolar hydatid disease]. / Interferonovyi status pri gel'mintozakh. Soobshchenie 2. Mnogokamernyi ékhinokokkoz.

In 11 patients with Echinococcus multilocularis infection the blood levels of alpha-interferon (alpha-IF) and gamma-interferon (gamma-IF) were measured using vesical stomatitis virus culture. In patients with the disease the contents of alpha- and gamma-IF were significantly (p less than 0.01) lower than in healthy controls: 392 +/- 101.6 and 101.6 +/- 20.9 U/ml and 1024 +/- 83.2 and 187 +/- 16.9 U/ml, respectively. The lower levels of alpha- and gamma-IF were found in patients with multiorgan damages including the lungs, as compared with the liver and abdomen infestation: alpha-IF 160 +/- 56.6, gamma-IF 88 +/- 24.0 and 525.7 +/- 124.3 and 109 +/- 18.3 (p less than 0.01), respectively. The similar data have been found in patients with hydatid disease (S. N. Suntsov et al., 1990). After 5 to 6 courses of mebendazole therapy in a dose of 50 mg/kg body weight dai y for 30 days with 30-day intervals there was a tendency to a slight elevation of cytokines levels (S. N. Suntsov et al., 1990), as seen in hydatid disease. The estimation of IF levels in multilocular echinococcosis infection may be of prognostic value and perhaps for the evaluation of the efficacy of surgical and chemotherapeutic treatment.

[The distribution of ABO system blood groups in patients with unilocular echinococcosis]. / Raspredelenie grupp krovi sistemy ABO u bol'nykh odnokamernym ékhinokokkozom.

The distribution of the AB0 blood groups was investigated in 624 patients with echinococcosis operated on and in 579 persons whose sera were subjected to IHAT with echinococcosis diagnosticum. Higher disease incidence was recorded in patients with the A (II) blood, especially in males, as compared to the controls. Simultaneous involvement of the liver, lungs and other organs was more common in them. Rh-positive patients predominated both in the A (II) and control groups. Blood group distribution with regard to IHAT findings failed to reveal any regular features.

Similarities of human hydatid cyst fluid components and the host serum.

Nine lung hydatid cyst fluid of Echinococcus granulosus species from man were analysed by electrophoresis, immunoelectrophoresis and biochemical tests. In addition respective sera of the host were analysed for comparison. Analysis revealed striking similarities in cellulose acetate and agarose gel electrophoretic and immunoelectrophoretic Patterns of proteins from hydatid cyst fluid and the serum of the respective hosts. It is presumed that serum proteins (albumins and globulins) occur in hydatid cyst fluid, but in small amount than in the host serum, and we believe that the host proteins can penetrate the membranes of the hydatid cyst.

[The level of the eosinophils in the blood, serum immunoglobulins and circulating immune complexes and interferon production in patients with unilocular echinococcosis depending on the location of the cysts]. / Soderzhanie éozinofilov v krovi, syvorotochnykh immunoglobulinov, tsirkuliruiushchikh immunnykh kompleksov i produktsiia interferonov u bol'nykh odnokamernym ékhinokokkozom v zavisimosti ot lokalizatsii kist.

The correlations between blood eosinophils, serum immunoglobulins (Ig), circulating immune complexes (CIC) content, alpha- and gamma-interferon (IFN) production and HLA class I antigens were analyzed in 33 patients with hydatid disease aged 37.5 +/- 1.5 years. Direct correlation between the activity of gamma-IFN production, blood eosinophils, IgG and specific IgE content was found in patients with multiple pulmonary damage. In this group the high level of CIC correlated with the high content of IgG and specific IgE. In patients with solitary and multiple cysts in the liver in spite of high alpha- and gamma-IFN production the levels of total and specific IgE were low, IgG and CIC contents were moderate and eosinophils percent was low. In patients with combined damage of the liver and multiple cysts in the abdominal cavity high CIC production correlated directly with high total IgE content. In the total group of patients percent of HLA B5 carriers was significantly higher than in controls: 36.6 and 14.7 respectively (R = 0.01 after Fischer) that correlated with comparatively active gamma-IFN production in patients with the pulmonary and the liver cysts. The possible explanation of the different immune response in patients with the different localisation of the parasite cysts are discussed.

[Agranulocytosis--a complication of the chemotherapy of echinococcosis with mebendazole]. / Agranulotsitoz--oslozhnenie khimioterapii ékhinokokkozov mebendazolom.

Only two of the 75 patients with echinococcosis, treated with mebendazole in a dose of 50 mg/kg b. m. for 30 days (single course) and administered from 1 to 13 courses with intervals of 1 to 3 months, developed agranulocytosis eventuating in death in one female patient. Analysis of blood levels of mebendazole and its metabolites permits a hypothesis that high concentrations thereof may have a toxic effect on the bone marrow, inducing its hypoplasia, in subjects sensitive to these substances.

Evaluation of ceruloplasmin levels in patients with pulmonary cystic echinococcus.

AIM: In this study, we aimed to evaluate serum CP levels in the serum samples before and after the surgical interventions in patients with pulmonary cystic echinococcus (CE). MATERIALS AND METHODS: Forty-eight patients with pulmonary CE who underwent surgery and 48 healthy individuals were enrolled to the study. Patients were divided in two groups; group 1 (n=48) consisted of patients with pulmonary CE, and group 2 (n=48) consisted of healthy subjects. Before and after surgical interventions serum CP levels were measured. RESULTS: Compared to group 2, group 1 had significantly higher CP levels at baseline (p <0.001). In group 1, CP levels were significantly decreased after the surgical intervention (p <0.001). CONCLUSIONS: The present study showed that CP levels increased in patients with pulmonary CE; chronic inflammation may cause these rises, and may be an immune response of the host, and these levels decreased after the surgical intervention.