Clonorchiasis is endemic in East Asia, and this zoonotic infection ranges from China, Hong Kong, Macao, and Korea, to Laos and Vietnam (Rim, 1986). There is a previous report that isolates of C. sinensis of Korea (Kimhae) and of China (Liaoning) showed low level intraspecific variation of DNA. The two populations are very closely related within the range of a genetic identity value of 0.998-1.0 and they have a high homology in the nucleotide sequences of the 18S rDNA, ITS2 (internal transcribed spacer 2) and cox1 (cytochrome c oxidase subunit 1) (Park and Yong, 2001). However, there had not been sequence data of ITS1 of C. sinensis. To understand whether the difference of the geographical region is related with the differences of the intraspecific variation, we compared the DNA sequences of C. sinensis from one Korean and two different Chinese isolates besides of Liaoning. DNA sequences of C. sinensis were analyzed for the nuclear rDNA (18S, ITS1 and ITS2) and mitochondrial DNA (cox1) to reveal how much the sequence variation is among the geographic isolates.Metacercariae of C. sinensis were collected after the artificial digestion of the muscle of the freshwater fish, Psudorasbora parva in Kimhae, Korea (A), Guangxi (B) and Shenyang (C), China. The adult worms were obtained from the rat (Sprague-Dawley, 4 to 6-weeksold male) liver one month post-infection with metacercariae (A and B) or from the rabbit liver five months post-infection (C). The worms collected were then stored at -70°C until assayed. The frozen worms were lyophilized and lysed with a lysis buffer containing 1% SDS, proteinase K (500 µg/ml), and RNase at 37°C for 2-3 hr. The DNA was extracted in phenol/ chloroform and precipitated in ethanol as reported by Abstract: We compared the DNA sequence difference of isolates of Clonorchis sinensis from one Korean (Kimhae) and two Chinese areas (Guangxi and Shenyang). The sequences of nuclear rDNA (18S, internal transcribed spacer 1 and 2: ITS1 and ITS2) and mitochondrial DNA (cytochrome c oxidase subunit 1: cox1) were compared. A very few intraspecific nucleotide substitution of the 18S, ITS1, ITS2 and cox1 was found among three isolates of C. sinensis and a few nucleotide insertion and deletion of ITS1 were detected. The 18S, ITS1, ITS2 and cox1 sequences were highly conserved among three isolates. These findings indicated that the Korean and two Chinese isolates are similar at the DNA sequence level.

We report 5 cases of ocular toxocariasis in Korean adults complaining of visual impairment along with floating or bubbling sensation. Fundoscopic examination revealed a retinal detachment along with exudate in 4 cases. They all showed typical reaction by ELISA and immunoblot against Toxocra excretory-secretory antigen. One case showed high level of anti-Toxocara IgE antibodies (34,000 Toxocara units/L) as well as increased level of serum total IgE antibodies and the specific IgE antibodies for 3 inhalant antigens, suggesting that high level of anti-Toxocara IgE antibodies was associated with an atopic status. Clinical manifestations were improved after the sequential use of steroids then mebendazole. We also suggest that ocular toxocariasis should be thoroughly investigated even when an evocative uniocular inflammatory lesion is encountered in peripheral retina without a systematic disease.

The validity of the CP2 gene of Cryptosporidium parvum as a viability marker was evaluated using absolute quantitative real-time polymerase chain reaction (qPCR) assays. Total ribonucleic acid (RNA) was isolated from live and heat-killed C. parvum oocysts, and complementary deoxyribonucleic acid was synthesized and used as a template. The most accurate number of viable C. parvum oocysts was predicted when the CP2 gene was used as a target gene. The lower detection limit of the CP2 gene was ten oocysts, which was the most sensitive among examined target genes. With heat shock induction, only hsp70 messenger RNA (mRNA) was induced, and the predicted viable oocyst number was increased by heat shock for this marker. The CP2, hsp70, Cryptosporidium oocyst wall protein, and beta-tubulin mRNAs were not detected in heat-killed oocysts, but the 18S ribosomal ribonucleic acid (rRNA) showed heat stability until 48 h after heat killing. Although the 18S rRNA demonstrated the fastest response in crossing point (CP) value among the examined primer sets in qPCR, overestimation of viable oocysts was noted in the analysis with this gene. In conclusion, the CP2 gene was identified as the most sensitive, reliable, and accurate candidate of a viability marker of C. parvum by qPCR evaluation.

Diabetes and obesity represent the major health problems and the most age-related metabolic diseases. Protein-tyrosine phosphatase 1B (PTP1B) has emerged as an important regulator of insulin signal transduction and is regarded as a pharmaceutical target for metabolic disorders. To find novel natural materials presenting therapeutic activities against diabetes and obesity, we screened various herb extracts using a chip screening allowing the determination of PTP1B inhibitory effects of the tested compounds using insulin receptor (IR) as the substrate. Cudrania tricuspidata leaves (CTe) had a strong inhibitory effect on PTP1B activity and substantially inhibited fat accumulation in 3T3-L1 cells. CTe was orally administrated to diet-induced obesity (DIO) mice once daily for 3 weeks after which changes in glucose, insulin metabolism, and fat accumulation were examined. Hepatic enzyme markers (aspartate aminotransferase, AST, and alanine aminotransferase, ALT) and total fat mass and triglyceride levels decreased in CTe-treated mice, whereas body weight and total cholesterol concentration slightly decreased. CTe increased the phosphorylation of IRS-1 and Akt in liver tissue. Furthermore, CTe treatment significantly lowered blood glucose levels and improved insulin secretion in DIO mice. Our results strongly suggest that CTe may represent a promising therapeutic substance against diabetes and obesity.

Pre-school children were examined for the presence of Enterobius vermicularis egg by perianal swab method. We visited 67 nurseries and 25 kindergartens between January, 1999 and April, 1999 in Chunchon, Korea. Of the 4,711 children examined, 434 (9.2%) were found to be positive for E. vermicularis egg. The egg positive rates of boys and girls were 10.1% and 8.1%, respectively. The rates were 7.8% in kindergartens and 9.7% in nurseries. Positive rate of 50 institutions was less than 10%. Rate in 35 institutions was from 10% to less than 20%. Rate in 7 institutions was equal to or over 20%. Out of 1,113 children examined twice, 28 (2.5%) children were positive consecutively, 53 (4.8%) were positive with negative conversion, 47 (4.2%) were negative with positive conversion, and 985 (88.5%) were consecutively negative. It means that in the low endemic area of enterobiasis with around 10% positive rate, the two consecutive examinations may increase the egg detection rate of 4.2-4.8%. The small number of consecutive egg positive children also suggests that the worm burden of the positive children might be low. Since the egg positive rate of that age group in this City increased from 1.85% (1997), and 3.0% (1998), the more intensive regular control should be executed.