• Journal of agricultural medicine and community health
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• v.22 no.2
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• pp.239-252
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• 1997

A questionnaire survey including stool examination for the parasite ova, skin test, and enzyme-linked immunosorbent assay (ELISA) of the liver fluke to find the factors related with the prevalence of the parasite infestation was done toward 118 residents living in three villages of Song-jung, Yi-jung, and Chim-gok of Okok-myun, Goksung County, South Cholla Province. The results are as follows : 1. The prevalence of Clonorchis sinensis ova by the stool examination was 46.6%. The sensitivity and specificity of skin test to egg-positive cases were 63.5% and 68.9%. The sensitivity and specificity of ELISA to egg-positive clonorchiasis were 100% and 34.9%. 2. The prevalence of Clonchiasis was higher in male than in female. 45~64 year old group was the highest among age groups, and the longer the duration of living in the present residence was, the higher the prevalence was shown statistically significantly (p<0.01). 3. The present drinker had higher prevalence of Clonorchiasis significantly than the past drinker or non-drinker, and the more the frequency of drinking the higher the prevalence was shown significantly (p<0.05). 4. Those who took raw fish had significantly higher than persons who didn't take it or take cooked fish(p<0.05) and prevalence showed the tendency to be increased as the frequency to take raw fishes increased significantly(p<0.05). 5. Those who had taken a raw sweet fish(Plecoglossus altivelis) or snake-headed fish(Ophicephalus argus) showed significantly higher prevalence of infection with liver fluke than persons who didn't take the raw fish. 6. While 56.4% of persons infected with Clonorchiasis answered that they were healthy, only 13.0% of the subjects were careful for their health, and 41.8% experienced tests to diagnose liver flukes before they were examined by this survey. 7. While 56.4% of infected persons thought the prevalence was high in their community, 54.0% were afraid to be infected, 47.1% thought the infection might destroy their health, 25.5% took treatment drugs, and 18.2% took raw fish continuously after treatment. The above results confirmed that Goksung area was highly prevalent in Clonorchiasis, so it was necessary to push effective preventive program against liver fluke infection together with the program to diagnose and treat existing infected persons.

• Parasites, Hosts and Diseases
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• v.57 no.1
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• pp.49-53
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• 2019

Contaminated liver fluke egg in the environment has led to the high prevalence of human opisthorchiasis associated with cholangiocarcinoma in Southeast Asia. To find the effective lessening methods of Opisthorchis viverrini eggs in the contaminated environment, we investigated the temperature conditions for killing of these trematode eggs in vitro. Numerous O. viverrini eggs were obtained in the proximal part of uteri of adult worms from experimental hamsters. Mature eggs with miracidium were allocated by experimental groups (2 control: positive and negative and 4 treatment: 50, 60, 70, and $80^{\circ}C$) with 0.85% saline, and treated by the experimental plan. Eggs in each experimental groups were observed under the confocal microscope after stain with Propidium Iodide (PI) to evaluate the effect of temperatures. Eggs in 70 and $80^{\circ}C$ groups were all killed after over 10 min heated. Majority of eggs in $60^{\circ}C$ (10, 15, and 30 min heated), 70 and $80^{\circ}C$ (5 min heated) groups were inactivated. However in $50^{\circ}C$ group, below half of eggs were to be killed in all time lapse (10, 15 and 30 min). In order to prevent O. viverrini infection and cholangiocarcinoma, direct treatment of sewage by heating at 70 or $80^{\circ}C$ at least 10 min is essential. Therefore, treatment of O. viverrini eggs at a high temperature is a potential method for controlling egg contamination in sewage.

• Korean Journal of Veterinary Research
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• v.41 no.4
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• pp.557-563
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• 2001

A field survey of fascioliasis of Korean native cattle raising and raised in specialized commercial breeding farms and local farms in Kangwon province using both intradermal test and sedimentation technique for feces was carried out from November to December, 1996. Fecal samples were taken from fascioliasis positive cattle by the intradermal test for the fecal examination. Liver tissues were randomly collected from an abattoir for histopathological examination of liver fluke infection in cattle. The results are as follows. 1. By the intradermal test for a total of 211 cattle raising in both Wonju and Wheongsung, Kangwon province, 60 heads(28.4%) showed positive reaction. Among 60 positive cattle, eggs of Fasciola hepatica were found from 51 heads(85.0%) by sedimentation technique. 2. According to the cattle raising areas, the positive rates by the intradermal test were 26.7%(20 out of 75 heads) in Wonju and 29.4%(40 out of 136 heads) in Wheongsung. 3. According to the age of cattle examined, the positive rates by the intradermal test in 1~3, 4~6 and 7~10 years old were 11.7%(7 out of 100 heads), 68.3%(41 out of 93 heads) and 20.0%(12 out of 18 heads), respectively. 4. The overall infection rates of fluke larvae from the slaughtered cattle at an abattoir in Wonju was 24.7%(37 out of 150 heads). In histopathology, liver lesions were observed such as inflammation with infiltration of eosinophils, polymorphonuclear cells, mononuclear cells and multinucleated giant cells, proliferation of connective tissues, calcification and abscess formation.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.1
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• pp.557-563
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• 2013

Intrahepatic cholangiocarcinoma (ICC) is ranked as one of the top five causes of cancer-related deaths. ICC in Thai patients is associated with infection with the liver fluke, Opisthorchis viverrini, but the molecular basis for development remains unclear. The present study employed a microarray approach to compare gene expression profiles of ICCs and normal liver tissues from the same patients residing in Northeast Thailand, a region with a high prevalence of liver fluke infection. In ICC samples, 2,821 and 1,361 genes were found to be significantly up- and down-regulated respectively (unpaired t-test, p<0.05; fold-change ${\gep}2.0$). For validation of the microarray results, 7 up-regulated genes (FXYD3, GPRC5A, CEACAM5, MUC13, EPCAM, TMC5, and EHF) and 3 down-regulated genes (CPS1, TAT, and ITIH1) were selected for confirmation using quantitative RT-PCR, resulting in 100% agreement. The metallothionine heavy metal pathway contains the highest percentage of genes with statistically significant changes in expression. This study provides exon-level expression profiles in ICC that should be fruitful in identifying novel genetic markers for classifying and possibly early diagnosis of this highly fatal type of cholangiocarcinoma.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.23
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• pp.10463-10468
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• 2015

Opisthorchis viverrini (Ov) infection is the major etiological factor for cholangiocarcinoma (CCA), especially in northeast Thailand. We have previously reported significant involvement of PI3K/AKT/PTEN and $Wnt/{\beta}$-catenin in human CCA tissues. The present study, therefore, examined the expression and activation of PI3K/AKT/PTEN and $Wnt/{\beta}$-catenin signaling components during Ov-induced cholangiocarcinogenesis in a hamster animal model. Hamsters were divided into two groups; non-treated and Ov plus NDMA treated. The results of immunohistochemical staining showed an upregulation of PI3K/AKT signaling as determined by elevated expression of the $p85{\alpha}$-regulatory and $p110{\alpha}$-catalytic subunits of PI3K as well as increased expression and activation of AKT during cholangiocarcinogenesis. Interestingly, the staining intensity of activated AKT (p-AKT) increased in the apical regions of the bile ducts and strong staining was detected where the liver fluke resides. Moreover, PTEN, a negative regulator of PI3K/AKT, was suppressed by decreased expression and increased phosphorylation during cholangiocarcinogenesis. We also detected upregulation of $Wnt/{\beta}$-catenin signaling as determined by increased positive staining of Wnt3, Wnt3a, Wnt5a, Wnt7b and ${\beta}$-catenin, corresponded with the period of cholangiocarcinogenesis. Furthermore, nuclear staining of ${\beta}$-catenin was observed in CCA tissues. Our results suggest the liver fluke infection causes chronic inflammatory conditions which lead to upregulation of the PI3K/AKT and $Wnt/{\beta}$-catenin signaling pathways which may drive CCA carcinogenesis. These results provide useful information for drug development, prevention and treatment of CCA.

• Parasites, Hosts and Diseases
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• v.50 no.4
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• pp.391-394
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• 2012

The prevalence of liver and intestinal fluke infections was determined by surveying inhabitants of Hengxuan, Fusui, and Shanglin villages which were known to be endemic for liver flukes in Guangxi, China in May 2010. A total of 718 people were examined for helminth eggs by the Kato-Katz thick smear technique, ultrasonography, immunoaffinity chromatography, and DNA sequencing. The overall egg positive rate was found to be 59.6% (28.0-70.6%) that included mixed infections with liver and intestinal flukes. Cases showing higher than 20,000 eggs per gram of feces (EPG) were detected between 1.3% and 16.2%. Ultrasonographic findings exhibited overall 28.2% (72 of 255 cases) dilatation rate of the intrahepatic bile duct. Clonorchis sinensis infection was detected serologically in 88.3% (38 of 43 cases) among C. sinensis egg positive subjects by the immunoaffinity chromatography using a specific antigen for C. sinensis. For differential diagnosis of the liver and intestinal flukes, more precise PCR and nucleotide sequencing for copro-DNA were performed for 46 egg positive cases. Mixed infections with C. sinensis and Metagonimus yokogawai were detected in 8 of 46 egg positive cases, whereas 29 specimens were positive for Haplorchis taichui. Ultrasonographic findings and immunoaffinity chromatography results showed usefulness, even in a limited way, in figuring out of the liver fluke endemicity.

• Parasites, Hosts and Diseases
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• v.55 no.3
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• pp.295-304
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• 2017

Opisthorchis viverrini infection induces chronic inflammation, and a minor proportion of infected individuals develop advanced periductal fibrosis (APF) and cholangiocarcinoma (CCA). Inflammatory cytokines and/or their gene polymorphisms may link to these biliary pathologies. We therefore investigated associations among cytokine gene polymorphisms and cytokine production in 510 Thai cases infected with O. viverrini who presented with APF+ or APF-, as established by abdominal ultrasonography as well as in patients diagnosed with CCA. Levels of pro-inflammatory and anti-inflammatory cytokines were determined in culture supernatants after stimulation of peripheral blood mononuclear cells (PBMCs) with O. viverrini excretory-secretory (ES) products. Pro-inflammatory cytokines, IL-$1{\beta}$, IL-6, IFN-${\gamma}$, LT-${\alpha}$, and TNF-${\alpha}$ were significantly increased in CCA patients compared with non-CCA (APF- and APF+) cases. Polymorphisms in genes encoding IL-$1{\beta}$-511C/T, IL-6-174G/C, IFN-${\gamma}$+874T/A, LT-${\alpha}$+252A/G, and TNF-${\alpha}$-308G/A were then investigated by using PCR-RFLP or allele specific-PCR (AS-PCR) analyses. In the CCA cases, LT-${\alpha}$+252A/G and TNF-${\alpha}$-308G/A heterozygous and homozygous variants showed significantly higher levels of these cytokines than the wild type. By contrast, levels of cytokines in wild type of IFN-${\gamma}$+874T/A were significantly higher than the variants in CCA cases. IFN-${\gamma}$+874T/A polymorphisms were associated with advanced periductal fibrosis, whereas IL-6-174G/C polymorphisms were associated with CCA. To our knowledge, these findings provide the first demonstration that O. viverrini infected individuals carrying several specific cytokine gene polymorphisms are susceptible to develop fibrosis and CCA.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.14
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• pp.5875-5881
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• 2015

Cholangiocarcinoma (CCA) is a rare but highly fatal cancer for which the molecular mechanisms and diagnostic markers are obscure. We therefore investigated the kinetic expression of isocitrate dehydrogenase-1 (IDH1), isocitrate dehydrogenase-2 (IDH2) and homogentisate 1,2-dioxygenase (HGD) during the tumorigenesis of O. viverrini infection-associated CCA in an animal model, and confirmed down-regulation of expression in human cases of opisthorchiasis-associated CCA through real time PCR. Kinetic expression of HGD, IDH1 and IDH2 in the animal model of O. viverrini infection-induced CCA was correlated with human CCA cases. In the animal model, expression of HGD was decreased at all time points (p<0.01) and expression of both IDH1 and IDH2 was decreased in the CCA group. In human cases, expression of HGD, IDH1 and IDH2 was decreased more than 2 fold in 55 cases (70.5%), 25 cases (32.1%) and 24 cases (30.8%) respectively. The present study suggests that reduction of HGD, IDH1 and IDH2 may be involve in cholangiocarcinoma genesis and may be useful for molecular diagnosis.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.5
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• pp.2963-2966
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• 2013

Infection with Opisthorchis viverrini (OV) due to eating certain traditional freshwater fish dishes is the principal risk factor for cholangiocarcinoma in Northeast Thailand where the infection is endemic and the incidence of this form of primary liver cancer has been the highest in the world. This paper is the second report of a prospective research project to monitor the impacts of a national liver fluke control programme in a rural community of Northeast Thailand. A sample of 684 villagers aged 20-65 years completed an interview questionnaire and were tested for infection using the Kato thick smear technique. The questionnaire was designed for the exploration of associations between OV infection, previous treatment with praziquantel, and knowledge and beliefs about the drug. The data were analysed using descriptive statistics and multiple logistic regression. The overall prevalence of OV infection was 37.2% and was highest in the 20-35 year age group, in those with a university degree and in those employed in the government sector. As many as 91.8% reported eating fish dishes known to place them at risk of infection. In the multiple regression analysis, previous use of praziquantel and lack of knowledge about whether or not the drug has a protective effect against re-infection were the only factors related to OV infection ($OR_{adj}$= 2.31, 95%CI =1.40-3.79 and $OR_{adj}$= 1.95, 95%CI= 1.24-3.05). The findings were discussed in terms of the possibly unwise dependency on praziquantel as a primary element in a control programme.

• Parasites, Hosts and Diseases
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• v.40 no.4
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• pp.191-193
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• 2002

Oriental liver fluke (Clonorchis sinensis) infection was surveyed among residents of Hamyang-gun, Gyeongsangnam-do, Korea during the period of January 2001 to March 2002. Total 1,041 stool samples were collected from residents who visited Public Health Center and its branches in Hamyang-gun and examined using formalin-ether sedimentation method. The overall egg positive rate was 16%, male showing higher positive rate (21%) than female (10%). The age group of 30 to 50 years had the highest egg positive rate of C. sinensis from 20% to 22%. The positive examinees were treated with praziquantel and educated individually to prevent reinfection . Egg positive rate in this area was decreased when compared with results recorded in the past, however, still remained more than 10%. This study suggests that periodic examination, treatment as well as education of residents should be continued and systematized.