Direct contact method of autoradiography was utilized in studying the distribution of exogenous C(14)-proline in Clonorchis sinensis, Eurytrema pancreaticum, Hymenolepis diminuta and Dipylidium caninum. The most distinct autoradiogram were obtained after 60 min of incubation, corresponding to the maximal absorption of C(14)-proline in these parasite. The radioactivitity of this labeled amino acid was chiefly concentrated in reproductive organs, especially egg-containing uterine tubules.
parasitology-helminth-trematoda-cestoda ; Clonorchis sinensis ; Eurytrema pancreaticum ; Hymenolepis diminuta ; Dipylidium caninum ; autoradiography ; C(14)-proline ; biochemistry ; amino acid

Clonorchis sinensis is a liver fluke and it is the most prevalent human parasite in Korea at present. The parasite infection induces immune responses, characteristically an increased production of parasite-specific IgE in the host. Major IgE-reacting C. sinensis antigens in infected humans have been protein bands with MWs of 15, 28, 37, 45, 51, 56, 62, 66, 74, 97 and 160 KD identified by immunoblot analysis. Individual variations of the IgE binding pattern to C. sinensis antigens have also been documented. Using immune BALB/c mouse sera, IgE-reacting protein bands have been visualized with MWs of 28, 74, 86, 160 and several > 200 KD. One of the most strongly reacted C. sinensis antigenic proteins with a molecular weight of 28 KD was purified by gel filtration and preparative electrophoresis. Using a monoclonal antibody produced against the antigenic protein, the protein was localized in the parasite's intestine, and also found to be contained in excretory-secretory antigens.
Animal ; Antibodies, Monoclonal ; Antigens, Helminth/immunology* ; Antigens, Helminth/analysis* ; Clonorchis sinensis/immunology* ; Female ; Fluorescent Antibody Technique ; Human ; IgE/immunology* ; Immunoblotting ; Mice ; Mice, Inbred BALB C

The nitric oxide (NO) formation and intrinsic nitrosation may be involved in the possible mechanisms of liver fluke-associated carcinogenesis. We still do not know much about the responses of inducible NO synthase (iNOS) induced by Clonorchis sinensis infection. This study was conducted to explore the pathological lesions and iNOS expressions in the liver of mice with different infection intensity levels of C. sinensis. Extensive periductal inflammatory cell infiltration, bile duct hyperplasia, and fibrosis were commonly observed during the infection. The different pathological responses in liver tissues strongly correlated with the infection intensity of C. sinensis. Massive acute spotty necrosis occurred in the liver parenchyma after a severe infection. The iNOS activity in liver tissues increased, and iNOS-expressing cells with morphological differences were observed after a moderate or severe infection. The iNOS-expressing cells in liver tissues had multiple origins.
Animals ; Clonorchiasis/*enzymology/genetics/parasitology/*pathology ; Clonorchis sinensis/*physiology ; Female ; Humans ; Liver/*enzymology/parasitology/pathology ; Mice ; Mice, Inbred BALB C ; Nitric Oxide Synthase Type II/*genetics/metabolism

This study was carried out to find out specific proteins from different organs of Clonorchis sinensis. Crude extract, organ-specific and excretory-secretory (ES) proteins were analyzed by immunoblot with infected human sera. The bands of 7- and 17-kDa were main component of intestinal fluid and ES protein and commonly found in all organspecific proteins. The 17-kDa protein was observed from ES antigen, intestinal fluid, eggs and sperms, 26- and 28- kDa proteins were from the uterus, vitellaria, and ovary, and 34-, 37-, 43- and 50-kDa proteins were mainly from the testis and sperms. Serum of mice immunized with sperms reacted to the 50-kDa protein by immunoblotting and immunohistochemical staining showed a positive reaction at the seminal receptacle and seminiferous tubule. The present results show that the 7-kDa protein is a common antigen of every part or organ of C. sinensis, but different organs express their specific antigenic protein bands.
Animals ; Antigens, Helminth/*analysis ; Clonorchis sinensis/anatomy & histology/*immunology ; Female ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Rabbits ; Research Support, Non-U.S. Gov't

Danggui, Agelicae Sinensis Radix, is a widely used Chinese herb to enrich blood, but its quality cannot be effectively assessed by the known chemical markers such as ferulic acid, ligustilide, polysaccharides, etc. A new bioassay was therefore developed to quantify the Enrich-Blood Bioactivity (EBB) for the quality assessment of Danggui raw materials. Danggui sample was first extracted with ethanol and water, respectively. Then the ethanolic extract and water extract were mixed as a test sample to quantify the amount of EBB by mice experiment. The blood deficiency mode in mice was developed by intraperitoneal injecting cyclophospharmide and phenylhdrazine hydrochloride. The quantity of red blood cell was chosen as EBB marker. Cyclosporine A was chosen as a control substance. EBB in analytes was quantified by the amount reaction of parallel line analysis (3, 3') method. The results indicated that the reliability test for quantifying EBB was passed through and the measured value was valid. The analytes showed the significant EBB (P < 0.05). The correlation coefficient was 0.9984 (n=5) between the amount of cyclosporine A (0.035-0.56 g x kg(-1)) and the increased number of red blood cell. The relative standard deviation (RSY) on the amount of EBB was estimated to be 6.15% (n = 6) by six replicated tests, and the confidence limit rate was 26.68% (n = 6). Five Danggui samples, which were collected from different cultivation areas with various morphological characters, showed the variety of EBB in the range of 21.95-44.16 U x g(-1). It is concluded that the developed method is accurate to quantify the EBB of Danggui raw materials, and is therefore suitable to assess its quality.
Angelica sinensis ; chemistry ; Animals ; Biological Assay ; methods ; Drugs, Chinese Herbal ; pharmacology ; Erythrocyte Count ; Erythrocytes ; drug effects ; Male ; Mice ; Mice, Inbred BALB C ; Plant Roots ; chemistry

We analyzed Clonorchis sinensis ancient DNA (aDNA) acquired from the specimens of the Joseon mummies. The target regions were cytochrome C oxidase subunit 1 (CO1), internal transcribed spacer 1 (ITS1), nicotinamide adenine dinucleotide hydrogen (NADH) dehydrogenase subunits 2 (NAD2) and 5 (NAD5). The sequences of C. sinensis aDNA was completely or almost identical to modern C. sinensis sequences in GenBank. We also found that ITS1, NAD2 and NAD5 could be good markers for molecular diagnosis between C. sinensis and the other trematode parasite species. The current result could improve our knowledge about genetic history of C. sinensis.
Clonorchis sinensis ; Cytochromes c ; Cytochromes ; Databases, Nucleic Acid ; Diagnosis ; DNA ; Electron Transport Complex IV ; Hydrogen ; Mummies ; NAD ; Niacinamide ; Oxidoreductases ; Parasites ; Republic of Korea

The mineral contents of the adult Clonorchis sinensis from rabbits and human were measured, and its qualitative and semi-quantitative analyses were studied with 3.4 meter Ebert-Grating spectrograph, and its quantitative analyses were studied with Beckman DU spectrophotometer. The adult Clonorchis sinensis used in this study were divided into two groups, the first group was collected from the bile passage of the man (C. sinensis from man), the second group was collected from the bile passage of the rabbit(C. sinensis from rabbit). Simultaneously, spectrographic and photometric analyses were also performed on the adult worms of Paragonimus westermani. Furthermore, comparative spectrographic analyses of the trace elements were carried out on the C. sinensis from man, C. sinensis from rabbit, Fasciola hepatica, Eurytrema pancreaticum, and Paragonimus westermani, and the approximate contents of the trace elements of the above trematodes were compared with that of their host tissues and biles. The results obtained were as follows: In the spectrographic analyses of C. sinensis from man, sodium, phosphorus, potassium, copper, calcium, aluminum, iron, and magnesium were detected predominantly, and zinc, titanium, silicone, lead, manganese, barium, cromium, molybdenum, and silver were detected as trace elements. In the differences of level of the minerals in these two C. sinensis, copper level of C. sinensis from man was higher than that of C. sinensis from rabbit, while calcium level of the former worm was higher than that of the latter worm. The trace level of lead, molybdenum, and silver were acertained in the former, but latter were not detected . The contents of the minerals showed the characteristic features in each trematodes: the mineral of flukes in each host were much more than that of the others; such as copper in C. sinensis from man, calcium in the C. sinensis from rabbit, and silicone in the P. westermani. The vanadium was detected in the F. hepatica, E. pancreaticum, and P. westermani, while the other flukes were not detected. In the comparative spectrographic analyses of the trace elements among the trematodes and their host tissues and biles, the minerals which detected from flukes were also found in their tissues and biles of their host. But the mineral levels of C. sinensis from man, F. hepatica, E. pancreaticum, and P. westermani were higher than that of their host tissues and biles, except the C. sinensis from rabbit.
parasitology ; helminth ; trematoda ; C. sinensis ; rabbit ; Fasciola hepatica ; Eurytrema pancreaticum ; Paragonimus westermani ; biochemistry ; sodium ; phosphorus ; potassium ; copper ; calcium ; aluminum ; iron ; magnesium ; zinc ; titanium ; silicone ; lead ; manganese ; barium ; cromium ; molybdenum ; silver ; vanadium

The 8 kDa antigenic protein of Clonorchis sinensis was partially purified by ammonium sulfate precipitation and subsequently by a column chromatographic steps. The purified protein was separated into 7 and 8 kDa protein bands through SDS-tricine gel electrophoresis, while the protein was found to migrate to a 8 kDa band in 7.5-15% SDS-PAGE. The molecular weight of the antigen was estimated to be 110 kDa by Superose 6 HR 10/30 gel filtration. The purified antigen strongly reacted with the human sera of clonorchiasis. The hyperimmune sera of BALB/c mice immunized against the 8 kDa protein were reacted with both the crude extract and the excretory-secretory product of adult worms, but not with the metacercarial extract. Immunohistochemical staining demonstrated that the protein was distributed to the tegument and subtegumental cells and also to the seminal receptacle. The present findings suggest that the 8 kDa protein is a partition of the multicomplex protein originating from various organs of adult C. sinensis, and that it is composed of several 7 and 8 kDa proteins.
Animals ; Antigens, Helminth/immunology/*isolation & purification/metabolism ; Clonorchiasis/immunology ; Clonorchis sinensis/anatomy & histology/*immunology/metabolism ; Helminth Proteins/immunology/*isolation & purification/metabolism ; Human ; Mice ; Mice, Inbred BALB C ; Molecular Weight ; Support, Non-U.S. Gov't

This study was undertaken to characterize the properties of a 100 kDa somatic antigen from Metagonimus yokogawai. Monoclonal antibodies (mAbs) were produced against this 100 kDa antigen, and their immunoreactivity was assessed by western blot analysis with patients' sera. The mAbs against the 100 kDa antigen commonly reacted with various kinds of trematode antigens, including intestinal (Gymnophalloides seoi), lung (Paragonimus westermani), and liver flukes (Clonorchis sinensis and Fasciola hepatica). However, this mAb showed no cross-reactions with other helminth parasites, including nematodes and cestodes. To determine the topographic distribution of the 100 kDa antigen in worm sections, indirect immunoperoxidase staining was performed. A strong positive reaction was observed in the tegumental and subtegumental layers of adult M. yokogawai and C. sinensis. The results showed that the 100 kDa somatic protein of M. yokogawai is a common antigen which recognizes a target epitope present over the tegumental layer of different trematode species.
Animals ; Antibodies, Helminth/immunology ; Antibodies, Monoclonal/*immunology ; Antigens, Helminth/*immunology ; Clonorchis sinensis/immunology ; Cross Reactions/immunology ; Fasciola hepatica/immunology ; Female ; Helminth Proteins/*immunology ; Heterophyidae/*immunology ; Immunologic Tests ; Mice ; Mice, Inbred BALB C ; Paragonimus westermani/immunology ; Trematode Infections/*diagnosis/immunology

BACKGROUND/AIMS: There are increasing numbers of North Korean defectors, and their health status differs from that of the South Korean population. This study was performed to investigate the clinical characteristics of North Korean defectors visiting a single tertiary hospital in South Korea. METHODS: The medical records of North Korean defectors who visited Dankook University Hospital, Cheonan, South Korea from 1 February 2006 to 30 April 2014 were retrospectively reviewed. Their comorbidities, main reasons for the visit, and most common diseases were investigated. RESULTS: A total of 169 North Korean defectors (163 females, 6 males) visited our hospital. Sixty-eight patients (40.24%) had comorbidities, the most common of which was tuberculosis (13.60%), followed by chronic hepatitis B (10.06%) and chronic hepatitis C (9.47%). North Korean refugees visited the department of ophthalmology (15.71%) most frequently, followed by gastroenterology (15.18%). Cataracts, chronic hepatitis C, chronic hepatitis B, and pulmonary tuberculosis were the most common reasons for the hospital visit. Chronic hepatitis B and chronic hepatitis C were diagnosed in 19.32% and 17.58% of the patients, respectively. Sixteen patients (9.47%) were diagnosed with tuberculosis, and eight of these patients showed multidrug resistance. Of all 169 patients, 17 underwent colonofibroscopy or stool examination, and parasites (Trichuris trichiura, n = 6; Clonorchis sinensis, n = 1) were found in 7 patients (41.18%). CONCLUSIONS: Most North Korean defectors who visited this tertiary hospital in South Korea were female, and they mainly visited the departments of ophthalmology, gastroenterology, and pulmonology. Compared with South Koreans, they showed high rates of chronic hepatitis B, chronic hepatitis C, pulmonary tuberculosis with multidrug resistance, and parasite infection.
Cataract ; Chungcheongnam-do ; Clonorchis sinensis ; Comorbidity ; Democratic People's Republic of Korea* ; Drug Resistance, Multiple ; Female ; Gastroenterology ; Hepatitis B, Chronic ; Hepatitis C, Chronic ; Humans ; Korea ; Medical Records ; Ophthalmology ; Parasites ; Pulmonary Medicine ; Refugees ; Retrospective Studies ; Tertiary Care Centers* ; Tuberculosis ; Tuberculosis, Pulmonary