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1.
Exp Ther Med ; 16(4): 3155-3160, 2018 Oct.
Article in English | MEDLINE | ID: mdl-30214538

ABSTRACT

Alveolar echinococcosis (AE) is a prevalent epidemic in the northern hemisphere, especially in central Europe and western China. Serum diagnosis is important for patients with AE, especially during the first screening. The present study purified the recombinant Em18-GST (rEm18-GST), and detected its diagnostic performance in human alveolar echinococcosis patients of Xinjiang, China with immunoblotting (IB) and enzyme-linked immunosorbent assay (ELISA). Serum samples were collected from 50 patients with AE, 222 patients with cystic echinococcosis (CE), 158 patients with other unrelated infections and 106 healthy individuals. The IB results showed that serum samples of 47 patients with AE and 12 patients with CE were rEm18-positive. However, only one sample from patients with cancer showed a cross-reaction with rEm18 in IB. The overall sensitivity was 94%, and the total specificity was 96.58%. For the rEm18 results using ELISA, the sera of 46 patients with AE were positive, and the overall sensitivity was 92%. In conclusion, compared with imaging tools, including computed tomography, magnetic resonance imaging and positron emission tomography, rEm18 has considerable advantages for AE serodiagnosis.

2.
Infect Dis Poverty ; 7(1): 61, 2018 Jun 13.
Article in English | MEDLINE | ID: mdl-29895318

ABSTRACT

BACKGROUND: Dogs play a pivotal role in the transmission of cystic echinococcosis (CE), a zoonosis caused by the tapeworm Echinococcus granulosus. We showed previously that dogs vaccinated with two E. granulosus adult-worm specific proteins, EgM9 and EgM123, emulsified with Freund's adjuvants induced significant protective efficacy in terms of reduction in worm burden and egg production after 45 days post-infection. It was not known whether this protection can be sustained using adjuvants suitable for use in dogs. METHODS: Recombinant EgM9 and EgM123 were mixed with Quil A or ISCOMs for vaccinating dogs. After three vaccine injections, all the dogs were orally challenge-infected with 200 000 protoscoleces of E. granulosus. After 45 days of infection, all the dogs were euthanized and necropsied for collecting and counting E. granulosus worms. Immunoglobins, including the IgG subclasses IgG1 and IgG2, were detected in the sera of vaccinated dogs by ELISA. To determine whether the protection efficacy could be maintained after 45 days post-infection, we implemented a longevity trial to count eggs in dog faeces for 170 days after infection. RESULTS: The dogs vaccinated with EgM9 and EgM123 mixed with Quil A and ISCOMs showed similar protective efficacy as the proteins emulsified with Freund's adjuvants in our previous study in terms of reduction of worms and eggs at 45 days post-infection. The longevity trial showed that EgM9 protein-vaccinated group released lower number of eggs per gram compared with the egg counts in the control dogs during the dog trial study. CONCLUSION: EgM9 and EgM123 are thus suitable vaccine candidates against E. granulosus infection in dogs.


Subject(s)
Echinococcosis/veterinary , Helminth Proteins/therapeutic use , Vaccination/veterinary , Vaccines/therapeutic use , Adjuvants, Immunologic/therapeutic use , Animals , Dogs , Echinococcosis/prevention & control , Echinococcus granulosus , Feces/parasitology , Female , Male , Parasite Egg Count/veterinary , Quillaja Saponins/therapeutic use , Recombinant Proteins/therapeutic use , Vaccines, Synthetic/therapeutic use
3.
BMC Cancer ; 17(1): 678, 2017 Oct 11.
Article in English | MEDLINE | ID: mdl-29020937

ABSTRACT

BACKGROUND: Diffusion-weighted MR imaging (DWI) has increasingly contributed to the management of nasopharyngeal carcinoma (NPC) patients. The objective of this paper was to explore the prognostic significance of apparent diffusion coefficient (ADC) values in 93 NPC patients. METHODS: This retrospective study included 93 newly diagnosed NPC patients. Pretreatment ADC values were determined and compared with patients' age, gender, alcohol intake, smoking, tumor volume, pathological type, tumor stage, and nodal stage. Using the Kaplan-Meier method, overall survival (OS), local relapse-free survival (LRFS), and distant metastasis-free survival (DMFS) were calculated and the values compared between the low and high ADC groups. Multivariate analysis of ADC values and other 9 clinical parameters was performed using a Cox proportional hazards model to test the independent significance for OS, LRFS and DMFS. RESULTS: The mean ADC value for the initial nasopharyngeal tumors was 0.72 × 10-3 mm2/s (range: 0.48-0.97 × 10-3 mm2/s). There was no significant difference between pretreatment ADCs and patient' gender, age, smoking, alcohol intake, or tumor stage. A significant difference in the ADCs for different N stages (P = 0.022) and correlation with initial tumor volume (r = -0.26, P = 0.012) were observed. In comparison, the ADC value for undifferentiated carcinoma was lower than that for other 3 pathological types. With a median follow-up period of 50 months, the 3-year and 5-year OS rates were 88.2% and 83.3%, respectively, 3-year and 5-year LRFS rates were 93.5% and 93.3%, respectively, and 3-year and 5-year DMFS rates were 83.9% and 83.3%, respectively. Patients with tumor ADC values ≥0.72 × 10-3 mm2/s exhibited longer OS and LRFS periods compared with tumor ADC values <0.72 × 10-3 mm2/s, with P values 0.036 and 0.018, respectively. In addition, patients with deaths or recurrences or distant metastasis had significant lower ADC values than those without disease failures. According to a multivariate analysis using the Cox proportional hazard test, ADC values showed a significant correlation with OS (P = 0.0004), LRFS (P = 0.0009), and DMFS (P < 0.0001), respectively. CONCLUSIONS: Pretreatment tumor ADC values supposed to be a noninvasive important prognostic parameter for NPC.


Subject(s)
Carcinoma/diagnostic imaging , Carcinoma/radiotherapy , Diffusion Magnetic Resonance Imaging/methods , Nasopharyngeal Neoplasms/diagnostic imaging , Nasopharyngeal Neoplasms/radiotherapy , Prognosis , Adult , Aged , Carcinoma/pathology , Disease-Free Survival , Female , Humans , Male , Middle Aged , Nasopharyngeal Carcinoma , Nasopharyngeal Neoplasms/pathology , Neoplasm Metastasis , Neoplasm Staging , Proportional Hazards Models , Radiotherapy, Intensity-Modulated , Tumor Burden
4.
Infect Dis Poverty ; 6(1): 88, 2017 May 03.
Article in English | MEDLINE | ID: mdl-28464914

ABSTRACT

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.


Subject(s)
Echinococcosis , Liver , Sheep Diseases , Animals , China , Echinococcosis/diagnostic imaging , Echinococcosis/radiotherapy , Echinococcosis/veterinary , Echinococcus granulosus , Female , Gene Expression Profiling , Liver/diagnostic imaging , Liver/metabolism , Liver/parasitology , Sheep , Sheep Diseases/diagnostic imaging , Sheep Diseases/radiotherapy , Tomography, X-Ray Computed , Treatment Outcome , Ultrasonography
5.
Article in Chinese | MEDLINE | ID: mdl-27089778

ABSTRACT

Echinococcosis is an important communicable disease that has remarkable impacts on the global health. The disease is highly endemic in western China. In the last decades, achievements were obtained for the surgery and drug therapies for echinococcosis, as well as for studies on genomics, signaling pathways, and liver proliferation and injury of the intermediate hosts. Although steps have entered vaccine development, challenges remainin immunodiagnosis and drug treatment for intermediate hosts, and in vaccine development for definitive hosts. This paper gives an overview on the current achievements and challenges for echinococcosis control.


Subject(s)
Echinococcosis , China , Humans , Infection Control
6.
Guang Pu Xue Yu Guang Pu Fen Xi ; 32(9): 2473-6, 2012 Sep.
Article in Chinese | MEDLINE | ID: mdl-23240420

ABSTRACT

Europium chloride, 2-thienylformyltrifluoroacetone and sodium silicate were used to synthesize new-style rare earth complex (Eu-TNS). By adding into dichloromethane solution containing Eu-TNS, the fluorescent intensities were enhanced gradually and regularly. High-resolution mass spectrometry was used to detect the formula of Eu-TNS, which belongs to multi-core rare-earth complex. Polarity of solution increasing by adding absolute ethanol will cause Eu-TNS to dissociate, which enhances the fluoresceot intensities of Eu-TNS solution. This rare earth complex Eu-TNS can be employed as fluorescence sensor to detect the content of ethanol in organic solvent.

7.
Zhonghua Yan Ke Za Zhi ; 48(3): 199-204, 2012 Mar.
Article in Chinese | MEDLINE | ID: mdl-22800416

ABSTRACT

UNLABELLED: OBJECTIVE To assess the prevalence of blindness and moderate and severe visual impairment among adults aged 50 years or above in the Shunyi district of Beijing,China. METHODS: It was a population-based cross-section study. Geographically defined cluster sampling was used in 5840 randomly selected individuals aged > or = 50 years in 28 clusters in the Shunyi district from April to August 2006. The survey was preceded by a pilot study where operational methods were refined and quality assurance evaluation was carried out. All participants were enumerated through village registers followed door-to-door visits. Eligible individuals were invited to receive visual acuity measurement and eye examination. Statistical analyses were performed using Stata/SE Statistical Software, release 9. 0. Chi-square test was used to investigate the association of age, gender and education with presenting and best corrected visual acuity. RESULTS: Five thousands eight hundreds and forty individuals were recruited and 5118 persons were examined,the response rate was 87.64%. Based on the criteria of World Health Organization visual impairment classification in 1973,57 persons were diagnosed as blindness, 160 persons were diagnosed as moderate and severe visual impairment defined as best corrected visual acuity, the prevalence of blindness and moderate and severe visual impairment were 1.11% and 3.12% respectively. Sixty-five persons were diagnosed as blindness, 428 persons were diagnosed as moderate and severe visual impairment defined as presenting visual acuity, the prevalence of blindness and moderate and severe visual impairment were 1.27% and 8. 36% respectively. The prevalence of blindness and moderate and severe visual impairment was higher in aged (trend Chi2 = 784.23, P = 0.000), female (Chi2 = 28.35, P = 0.000), and illiterate persons (trend Chi2 = 397.13, P = 0.000). Cataract was still the first leading cause of blindness and visual impairment, the retinal diseases was the second. Un-corrected refractive error also was the main cause of visual impairment. CONCLUSIONS: The prevalence of blindness and moderate and severe visual impairment has been decreasing among older adults aged 50 years or above in the Shunyi district since 1996. The first leading cause of blindness and visual impairment is still cataract.


Subject(s)
Blindness/epidemiology , Vision, Low/epidemiology , Age Distribution , Aged , Cataract/epidemiology , China/epidemiology , Cross-Sectional Studies , Female , Humans , Male , Middle Aged , Prevalence
8.
Zhonghua Yan Ke Za Zhi ; 48(3): 211-8, 2012 Mar.
Article in Chinese | MEDLINE | ID: mdl-22800418

ABSTRACT

OBJECTIVE: To assess the prevalence of cataract, the outcomes of cataract surgery, the rate of cataract surgical coverage, and the rate of cataract blindness social burden. METHODS: Cluster sampling was used in 5118 randomly selected individuals aged > or = 50 years in the Shunyi district of Beijing from April to August 2006. All individuals were received visual acuity and eye examination, including the evaluation of lens status and cataract surgical status by slit lamp biomicroscopy. Data bank was established by Epi-info Software. Statistical analyses were performed using Stata/SE Statistical Software, release 9.0 (Stata Corp, College Station,TX). Multiple logistic regression analysis was employed to analyzed the prevalence of cataract,cataract surgical coverage rate,cataract blindness social burden rate and outcomes of surgery among different ages, sex and educational background. RESULTS: In 5118 individuals, 797 cases of cataract were found, the prevalence of cataract was 15.57% among adults aged 50 or above. The prevalence of cataract was higher in the aged, female, illiterate persons (P < 0.01). In 189 eyes with cataract surgery, 65.08% and 23.28% of eyes were performed by the modern extra-capsular surgery and Phacoemulsification respectively. The rate of intraocular lens implantation was 89.42%. Post-operative presenting and best corrected visual acuity more than 0.7 was 30.16% and 61.38% of operated eyes respectively. The main causes of the post-operated eyes with worse visual acuity were post-capsular opacity and refractive error. The cataract surgical coverage rate was 62.22%, and the cataract blindness social burden rate was 2.64%. The cataract surgical rate was lower and cataract blindness social burden rate was higher in the aged persons (P < 0.01). CONCLUSIONS: Cataract is the most common eye disease that may lead into blindness and severe visual impairment among older adults aged equal or more than 50 years. The cataract surgical coverage rate is high in the Shunyi district, but still is lower in the aged persons. The visual outcomes of the surgery should be further improved in the future.


Subject(s)
Cataract Extraction/statistics & numerical data , Cataract/epidemiology , Cataract/therapy , Age Distribution , Aged , Aged, 80 and over , China/epidemiology , Female , Humans , Male , Middle Aged , Prevalence
9.
Guang Pu Xue Yu Guang Pu Fen Xi ; 31(3): 727-31, 2011 Mar.
Article in Chinese | MEDLINE | ID: mdl-21595228

ABSTRACT

Eu(PPA)3 dioxane and Tb(PPA)3 dioxane were synthesized by firstly adopting dioxane as second ligand and using pipemidic acid (PPA) as first ligand. In contrast with two kinds of binary lanthanide complexes Eu(PPA)3 and Tb(PPA)3, the photoluminescent intensities of Eu (PPA)3 dioxane and Tb (PPA)3 dioxane are much stronger, especially for that of Tb(PPA)3 dioxane. The enhancement of photoluminescent intensities of Eu(PPA)3 dioxane and Tb(PPA)3 dioxane is because of the replacement of dioxane for water bonded to Eu3+ and Tb3+, by which the energy loss through heat oscillation will be inhibited. The as-synthesized lanthanide complexes were characterized by FT-IR, photoluminescence spectra and photoluminescence lifetime. The lifetime of Eu(PPA)3 dioxane and Tb(PPA) dioxane remarkably changed compared with those of Eu(PPA)3 and Tb(PPA)3. However, there is much difference for the lifetime change of as-synthesized lanthanide complexes formed by Eu3+ and Tb3+, which shows that there is different energy transfer process for the lanthanide complexes formed by Eu3+ and Tb3+. The synthesis of lanthanide complexes using dioxane as second ligand will provide a new method to detect the existence of dioxane.

10.
Article in Chinese | MEDLINE | ID: mdl-20066992

ABSTRACT

OBJECTIVE: To investigate the immune protection of the recombinant membrane protein(45M) against larvae of Taenia multiceps in sheep. METHODS: Twelve sheep were randomly divided into immune group and control group, and each sheep was immunized four times (three weeks interval) respectively with 50 microg recombinant 45M or GST emulsified with adjuvant. Serum samples were collected at pre-immunization and certain time after immunization, and were analyzed for IgG and IgM by ELISA. At the 105th day after the final inoculation, each sheep was orally challenged by 5000 eggs of T. multiceps. Two weeks after infection, the sheep were sacrificed, serum samples were collected, and number of cysts in brain tissue was counted. Activated oncospheres were cultured with 10% sera of immune group or control group, and observed by light microscopy. RESULTS: The mean number of cysts was 1.5, and the average diameter of the cysts was 2.2 mm. Compared with control group, the reduction rate of cysts in immunized sheep was 68.9%. At the 72th hour after culture with serum of immunized sheep, about 90% oncospheres were killed by antiserum, and the oncospheres began to shrink or expand, and internal structure became fuzzy. ELISA showed that after final immunization (the 9th week), the levels of IgG in sera of immune group (2.32+/-0.76) were significantly higher than those of the control (0.70+/-0.42) (t=4.47, P<0.01). At the 24th week the levels of IgG and IgM in sera of immune group (1.53+/- 0.81, 0.90+/-0.26) were significantly higher than those of the control (0.64+/-0.43, 0.43+/-0.15) (P<0.01). CONCLUSION: The recombinant 45M can induce certain humoral immune response.


Subject(s)
Cysticercosis/veterinary , Membrane Proteins/immunology , Sheep Diseases/immunology , Taenia/immunology , Animals , Antibodies, Helminth/blood , Antibody Formation , Cysticercosis/immunology , Female , Male , Recombinant Proteins/immunology , Sheep , Sheep Diseases/prevention & control
11.
Article in Chinese | MEDLINE | ID: mdl-24818364

ABSTRACT

OBJECTIVE: To verify the application and effectiveness of monthly deworming for all dogs in the control of Echinococcus granulosus infection. METHODS: Baited praziquantel was used to treat all registered dogs monthly by hydatid disease control officers at village level and all stray (unowned) dogs were eliminated in the counties of Hutubi and Wensu in 1987-1990 and 1990-1994 respectively. Prevalence of echinococcosis in dogs and sheep was recorded yearly. RESULTS: The infection rate in dogs decreased from 18.5% and 14.7% before implementation of the control measure to zero in 3-4 years in Hutubi and Wensu Counties respectively. Prevalence of hydatid disease in new born sheep was reduced by more than 85% in comparison to the same age sheep before the control program in both counties. CONCLUSION: "Monthly drug administration to all dogs" is an effective way to the control of echinococcosis in dogs and of hydatid disease in sheep.


Subject(s)
Dog Diseases/prevention & control , Dog Diseases/parasitology , Echinococcosis/veterinary , Animals , China , Dogs , Echinococcosis/prevention & control , Sheep , Sheep Diseases/parasitology , Sheep Diseases/prevention & control
12.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 23(11): 998-1000, 2007 Nov.
Article in Chinese | MEDLINE | ID: mdl-17988575

ABSTRACT

AIM: To study the localization of the antioxidant protein thioredoxin peroxidase (TPx) of Echinococcus granulosus (EgTPx) in the protoscoleces (PSC) of parasite. METHODS: The protoscoleces of E.granulosus were aspirated and pooled from sheep liver hydatid cysts. After digested by pepsin, the sedimented protoscoleces were used for paraffin sections. The localization of the antioxidant protein TPx of EgTPx was determined by using(polyclonal antibody against rEgTPx) and indirect immunofluorescence staining. RESULTS: Indirect immunofluorescence staining analysis indicated that the rEgTPx was mainly distributed in tegument, subtegument and calcareous corpuscle cells of the protoscolex. CONCLUSION: The wide distribution and large sites of EgTPx in the parasite hare been clearly determined, which will help further investigation into the biological functions and application of TPx protein.


Subject(s)
Antioxidants/analysis , Echinococcus granulosus/enzymology , Peroxiredoxins/metabolism , Animals , Echinococcus granulosus/cytology , Echinococcus granulosus/metabolism , Fluorescent Antibody Technique, Indirect , Immune Sera/immunology , Mice , Peroxiredoxins/analysis , Peroxiredoxins/immunology
13.
Zhongguo Zhong Xi Yi Jie He Za Zhi ; 27(7): 642-4, 2007 Jul.
Article in Chinese | MEDLINE | ID: mdl-17717929

ABSTRACT

OBJECTIVE: To comparatively observe the curative effect, adverse reaction and compliance of Modified Xiaoyao Pill combining amitriptyline (MXP-At) in treating patients with depression. METHODS: Sixty-four patients with diagnosis of depression matched to the Chinese Classification of Mental Disorders (CCMD-3) were randomly assigned to 2 groups, the treatment group treated with MXP-At and the control group with fluoxeline, 32 cases in each group. The curative effect was evaluated by Hamilton depression (HAMD) scale and the adverse reaction was recorded before treatment and at the 2nd, 4th and 12th week of the treatment. Patients were regularly followed up from the 12th week to the 24 th week. The curative effect and compliance in the two groups were compared. RESULTS: The HAMD score dropped in both groups from the 2nd week of the treatment, and at that time, it was lower in the control group than that in the treatment group (P < 0.05); but at the 4th week, no significant difference was found in the therapeutic effect and the HAMD score between the two groups. However, 3 and 14 cases in the treatment and the control group were relapsed during the 12 weeks of follow-up respectively. CONCLUSION: MXP-At shows a curative effect similar to fluoxetine on depression but with less adverse reaction, and is not expensive.


Subject(s)
Amitriptyline/therapeutic use , Depression/drug therapy , Drugs, Chinese Herbal/therapeutic use , Adolescent , Adult , Antidepressive Agents, Tricyclic/therapeutic use , Drug Therapy, Combination , Female , Follow-Up Studies , Humans , Male , Middle Aged , Patient Compliance/statistics & numerical data , Phytotherapy , Treatment Outcome , Young Adult
14.
Zhonghua Yan Ke Za Zhi ; 42(11): 980-4, 2006 Nov.
Article in Chinese | MEDLINE | ID: mdl-17386134

ABSTRACT

OBJECTIVE: To investigate a rat animal model of filtering bleb scarring formation. METHODS: Thirty-five of Sprague-Dawley (SD) rats were used in this experiment. The filtration surgery was performed on one eye of each rat and the other eye served as control. The presence of postoperative bleb and clinical manifestation was evaluated. Each group of three rats were sacrificed and the eyes enucleated on day 1, 3, 7, 14, and 28. After the eyes were fixed by 10% formalin, serial paraffin sections of three eyes were labeled with HE staining and the pathological morphology was observed under a microscope. RESULTS: A conjunctival bleb was successfully formed in 35 rats lasting from 8 to 21 days after surgery. An obvious dilatation of blood vessels, infiltration of a great number of neutrophil, and a severe edema of the subconjunctival tissue were observed on day 1 and the above morphological changes plus increased infiltration of macrophages were found on day 3. The dilated blood vessels were lessened while the proliferation of fibroblast was presented with abundant endochylema in oval-shaped nucleus of fibroblasts, which were closely associated with the deposition of small amount of new loose collagen fibers on day 7. The collagen fibers became tight on day 14 and subsequently, the early scar was formed on day 28. CONCLUSION: The rat model of filtering bleb scarring formation may be useful for the investigation of preventing and treating conjunctival scarring resulted from glaucoma filtration surgery and other related eye diseases in the future.


Subject(s)
Cicatrix , Conjunctival Diseases , Filtering Surgery/methods , Animals , Cicatrix/pathology , Conjunctival Diseases/pathology , Disease Models, Animal , Rats , Rats, Sprague-Dawley
15.
Exp Parasitol ; 110(1): 88-90, 2005 May.
Article in English | MEDLINE | ID: mdl-15804383

ABSTRACT

We describe a method for obtaining improved secondary infections of Echinococcus granulosus that involves culturing protoscolex larvae in vitro prior to inoculation into mice. This approach provides a far superior method for obtaining secondary echinococcosis infections in mice compared with the traditional method of direct inoculation of protoscoleces (PSC) where the majority of parasites are killed by the host. We obtained a high rate of recovery both in terms of secondary cyst numbers and their viability. After 50 weeks post-infection (p.i.), brood capsules were formed and the first PSC developed in each of the capsules. After 56 weeks p.i., the fastest developing brood capsule contained four PSC. The approach will prove valuable for investigating parasite development and the host-parasite interaction in secondary echinococcosis.


Subject(s)
Echinococcosis/parasitology , Echinococcus granulosus/growth & development , Animals , Echinococcosis/immunology , Echinococcus granulosus/immunology , Echinococcus granulosus/pathogenicity , Host-Parasite Interactions , Mice , Mice, Inbred BALB C
16.
Biol Proced Online ; 6: 67-77, 2004.
Article in English | MEDLINE | ID: mdl-15188015

ABSTRACT

Three cDNAs, termed EpC1, TPxEg and EgG5, were isolated by immunoscreening from an Echinococcus granulosus cDNA library. The recombinant phages exhibited strong reactivity with sera from humans with confirmed cystic echinococcosis (CE) and with sera from mice infected with E. granulosus oncospheres. The cDNAs were subcloned into a pET vector, expressed as fusion proteins tagged with GST and affinity purified against the GST tag. Of the three recombinant proteins, EpC1 achieved the highest performance for serodiagnosis of CE in Western blot analysis using a panel of clinically defined human sera to initially address the sensitivity and specificity of the molecules. The protein yielded an overall sensitivity of 92.2% and specificity of 95.6%, levels unprecedented taking into account the large panel of 896 human sera that were tested. The strategy used may also prove suitable for improved immunodiagnosis of other parasitic infections.

17.
Gene ; 326: 157-65, 2004 Feb 04.
Article in English | MEDLINE | ID: mdl-14729274

ABSTRACT

A full-length cDNA sequence coding for Echinococcus granulosus thioredoxin peroxidase (EgTPx) was isolated from a sheep strain protoscolex cDNA library by immunoscreening using a pool of sera from mice infected with oncospheres. EgTPx expressed as a fusion protein with glutathione S-transferase (GST) exhibited significant thiol-dependent peroxidase activity that protected plasmid DNA from damage by metal-catalyzed oxidation (MCO) in vitro. Furthermore, the suggested antioxidant role for EgTPx was reinforced in an in vivo assay, whereby its expression in BL21 bacterial cells markedly increased the tolerance and survival of the cells to high concentrations of H2O2 compared with controls. Immunolocalization studies revealed that EgTPx was specifically expressed in all tissues of the protoscolex and brood capsules. Higher intensity of labelling was detected in many, but not all, calcareous corpuscle cells in protoscoleces. The purified recombinant EgTPx protein was used to screen sera from heavily infected mice and patients with confirmed hydatid infection. Only a portion of the sera reacted positively with the EgTPx-GST fusion protein in Western blots, suggesting that EgTPx may form antibody-antigen complexes or that responses to the EgTPx antigen may be immunologically regulated. Recombinant EgTPx may prove useful for the screening of specific inhibitors that could serve as new drugs for treatment of hydatid disease. Moreover, given that TPx from different parasitic phyla were phylogenetically distant from host TPx molecules, the development of antiparasite TPx inhibitors that do not react with host TPx might be feasible.


Subject(s)
Antioxidants/metabolism , Echinococcus/genetics , Gene Expression Regulation, Enzymologic , Peroxidases/genetics , Animals , Blotting, Western , Cloning, Molecular , DNA, Complementary/chemistry , DNA, Complementary/genetics , Echinococcus/enzymology , Echinococcus/growth & development , Escherichia coli/drug effects , Escherichia coli/genetics , Escherichia coli/growth & development , Gene Expression Regulation, Developmental , Glutathione Transferase/genetics , Glutathione Transferase/metabolism , Humans , Hydrogen Peroxide/pharmacology , Immune Sera/immunology , Molecular Sequence Data , Peroxidases/immunology , Peroxidases/metabolism , Peroxiredoxins , Phylogeny , RNA, Helminth/genetics , RNA, Helminth/metabolism , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis, DNA
18.
J Infect Dis ; 188(12): 1951-60, 2003 Dec 15.
Article in English | MEDLINE | ID: mdl-14673776

ABSTRACT

A pool of serum samples from mice infected with oncospheres (eggs) of Echinococcus granulosus was used to screen a cDNA library constructed with RNA extracted from protoscolex larvae from sheep hydatid cysts. One immunoreactive clone, designated EpC1, was shown to encode a protein of 76 residues. The complementary DNA (cDNA) fragment was subcloned into an expression vector, pET-41b(+), and the resulting recombinant EpC1 glutathione S-transferase (GST) fusion protein (rEpC1-GST) was expressed in Escherichia coli and was affinity purified against the GST tag. Immunoglobulin G was the dominant antibody isotypes generated against rEpC1-GST. A total of 896 human serum samples were used to evaluate the diagnostic sensitivity and specificity of the fusion protein by immunoglobulin G immunoblotting; 324 serum samples from patients with cystic echinococcosis (CE), 172 from patients with neurocysticercosis, 89 from patients with alveolar echinococcosis, and 241 from patients with other infections or clinical presentations, as well as 70 from confirmed-negative control subjects, yielded an overall sensitivity of 92.2% and an overall specificity of 95.6%. The combined levels of sensitivity and specificity achieved with the rEpC1-GST fusion protein for diagnosis of CE are unprecedented, taking into account the large panel of serum samples that were tested.


Subject(s)
Antibodies, Helminth/blood , Antigens, Helminth/biosynthesis , Echinococcosis/diagnosis , Echinococcus/isolation & purification , Helminth Proteins/biosynthesis , Immunoblotting , Recombinant Fusion Proteins/biosynthesis , Amino Acid Sequence , Animals , Antigens, Helminth/genetics , Antigens, Helminth/immunology , Cloning, Molecular , Cross Reactions , Disease Models, Animal , Echinococcosis/blood , Echinococcosis/surgery , Echinococcus/genetics , Echinococcus/immunology , Enzyme-Linked Immunosorbent Assay , Escherichia coli/metabolism , Female , Genetic Vectors , Glutathione Transferase/genetics , Helminth Proteins/genetics , Helminth Proteins/immunology , Humans , Immunoglobulin G/blood , Male , Molecular Sequence Data , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/immunology , Sensitivity and Specificity , Sequence Alignment
19.
Article in Chinese | MEDLINE | ID: mdl-14628351

ABSTRACT

OBJECTIVE: To study expression and sequence differences of Echinococcus granulosus 95(Eg95) antigen cDNA from different stages of protoscolex, oncosphere and adult worm of E. granulosus from Xinjiang Uighur Aut. Reg. METHODS: In accordance with the sequence of Eg95 antigen cDNA, the primers of Eg95 were designed. Eg95 antigen cDNAs were amplified by PCR from protoscolex, oncosphere and adult worm cDNA libraries of E. granulosus, respectively and were cloned into pUCm-T plasmid, and sequenced. The sequences were analyzed by DNAman and GenBank/BLAST biosoftware. RESULTS: PCR results showed that Eg95 antigen cDNA was amplified from three stages of E. granulosus cDNA libraries. Sequencing analysis indicated that the Eg95 cDNA length was 402 bp, same as the reported data in GenBank. CONCLUSION: The Eg95 antigen cDNA was expressed in the different life-cycle stages of E. granulosus in Xinjiang and there was no nucleic acid sequence difference of Eg95 antigen among the protoscolex, oncosphere and adult worm of E. granulosus.


Subject(s)
Antigens, Helminth/genetics , DNA, Complementary/genetics , Echinococcus/genetics , Helminth Proteins/genetics , Animals , Antigens, Helminth/chemistry , Base Sequence , Cloning, Molecular , DNA, Complementary/chemistry , Echinococcus/immunology , Helminth Proteins/chemistry , Polymerase Chain Reaction , Sequence Homology, Nucleic Acid
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