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Characterization of Ferritin Isolated from Dog Spleen (개의 비장에서 분리한 페리틴의 특성)

  • Park Jae-Hag;Jun Do Youn;Kim Young Ho
    • Journal of Life Science
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    • v.15 no.3 s.70
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    • pp.439-446
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    • 2005
  • Ferritin is known to be the principle iron-storage protein in a wide variety of rganisms. The electro­phoretic mobility and immunological cross-reactivity of dog splenic ferritin were compared with those of horse, bovine, and pig splenic ferritin after isolation using heat treatment, salting out, column chromatography, and ultrafiltration. These isolation methods allowed the recovery of $\~84{\mu}g$ of the ferritin per g of spleen. The iron content in the dog ferritin was $22.7\%$, which appeared to be higher than those in the other mammalian ferritins tested. The electrophoretic mobility of the dog ferritin under nondenaturing conditions was similar to its bovine counterpart, whereas it was more identical to pig and horse ferritins on an SDS-polyacrylamide gel. The molecular weight of the dog ferritin subunit was 19.5 kDa on an SDS-polyacrylarnide gel, and the subunit was unable to bind with iron. The polyclonal anti-dog ferritin raised in rats was able to cross-react with the pig, bovine, and horse ferritins, upon Ouchterlony double immunodiffusiion. A Western blot analysis also revealed that the anti-dog ferritin, which specifically bound with the dog ferritin subunit, could also recognize the horse, bovine, and pig ferritin subunits and the maximum cross-reactivity was exhibited with the pig ferritin subunit, indicating that the dog ferritin is immunochemically more similar to the pig ferritin than its other mammalian counterparts. Accordingly, these results elucidate the biochemical and immunochemical characteristics of dog ferritin that might have a potential to be applied as an oral iron supplement to treat iron deficiency anemia.

Novel Real Time PCR Method for Detection of Plasmodium vivax (새로운 Real Time PCR 방법을 통한 Malaria(Plasmodium vivax)의 검출)

  • Ki, Yeon-Ah;Kim, So-Youn
    • Microbiology and Biotechnology Letters
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    • v.33 no.2
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    • pp.148-153
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    • 2005
  • Malaria is a re-emerging infectious disease that is spreading to areas where it had been eradicated, such as Eastern Europe and Central Asia. To avoid the mortality from malaria, early detection of the parasite is a very important issue. The peripheral blood smear has been the gold standard method for the diagnosis of malaria infection. Recently, several other methods have been introduced for quantitative detection of malaria parasites. Real time PCR that employs fluorescent labels to enable the continuous monitoring of PCR product formation throughout the reaction has recently been used to detect several human malaria parasites. 18S rRNA sequences from malaria parasites have been amplified using Taqman real time PCR assay. Here, a SYBR Green-based real time quantitative PCR assay for the detection of malaria parasite-especially, Plasmodium vivax - was applied for the evaluation of 26 blood samples from Korean malaria patients. Even though SYBR Green-based real time PCR is easier and cheaper than Taqman-based assay, SYBR Green-based assay cannot be used because 18S rRNA cannot be specifically amplified using 1 primer set. Therefore, we used DBP gene sequences from Plasmodium vivax, which is specific for the SYBR Green based assays. We amplified the DBP gene from the 26 blood samples of malaria patients using SYBR Green based assay and obtained the copy numbers of DBP genes for each sample. Also, we selected optimal reference gene between ACTB and B2M using real time assay to get the stable genes regardless of Malaria titer. Using selected ACTB reference genes, we successfully converted the copy numbers from samples into titer, ${\sharp}$ of parasites per microliter. Using the resultant titer from DBP based SYBER Green assay with ACTB reference gene, we compared the results from our study with the titer from Taqman-based assay. We found that our results showed identical tendency with the results of 18S rRNA Taqman assay, especially in lower titer range. Thus, our DBP gene-utilized real time assay can detect Plasmodium vivax in Korean patient group semi-quantitatively and easily.

A Comparison of the Disease and Insect Damage, and Growth Characteristics of Sorghum and Foxtail Millet between Rotational Upland and Continuously Cropped Upland Field in Order to Identify the Optimum Varieties for an Organic Paddy-Upland Rotation System (답전윤환 적합 품종 선발을 위한 윤환밭과 연속밭에서의 수수, 조의 병충해, 생육 특성 비교)

  • Yu, Je-Bin;Yoon, Seong-Tak;Yang, Jing;Ye, Min-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.62 no.3
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    • pp.224-232
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    • 2017
  • In this study, we investigated the disease, insect damage, growth, and yield characteristics of sorghum and foxtail millet in an organic paddy-upland rotation system at Anseong-si Gyeonggi province, Korea, in 2016. Seven varieties of sorghum and foxtail millet are used in this study. A rotational upland field and continuously cropped upland field were compared in order to establish an optimum cultural method and to select the best sorghum and foxtail varieties for a paddy-upland rotation system. The incidence rate of major diseases was higher for sorghum than for foxtail millet. Moktaksusu (22.8%) had the highest incidence rate among sorghum varieties. Hwanggeumchal (7.0%) had the highest incidence rate among foxtail millet varieties. DS202 (89.3%) was the most severely damaged by Ostrinia furnacalis larva, which is the main pest of sorghum and maize. The weed numbers, and the weed fresh and dry weights were lower in the rotational upland field than in the continuously cropped upland field. Futhermore, the number of weed plants decreased by 42% and the dry weight of weeds decreased by 33% in the rotational upland field compared to the continuously cropped upland field. The culm length of sorghum and foxtail millet were 7.9 cm, and 3.4 cm longer, respectively, in the rotational upland field than in the continuously cropped upland field. The average yield of sorghum per 10 a increased by 90% in the rotational upland field compared to continuously cropped upland field. However, there was no significant difference between the rotational upland field and the continuously cropped upland field for foxtail millet. The most suitable varieties for a paddy-upland rotation system were judged to be Hwanggeumchal, Donganmae, and DS202 for sorghum, and Samdachal, Samdamae, Dahwangmae for foxtail millet.

Study on the Food Habits of Korean Native Goats (Capra hircus) Fed with Various Roughage Sources (여러 가지 조사료를 급여한 재래산양의 채식습성에 관한 연구)

  • Lee, In-Duk;Lee, Hyung-Suk
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.28 no.2
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    • pp.119-128
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    • 2008
  • The object of this experiment was to investigate the food habits of Korean native goats fed with various roughage sources. The feeding trials were conducted at Chungnam National University and Unbong Animal Genetic Resources Station in 2007. The experimental roughages sources were five including 25 species in all; grass sources: 5 species (Lolium perenne, Festuca arundinacea, Poa pratensis, Agrostis alba and mixed grass, legume sources: 5 species (Trifolium pratense, Medicago sativa, Trifolium repens, Melilotus officinalis and Vicia villosa), browse sources: 5 species(Quercus serrata Thunb., Prunus jamasakura Sieb., Quercus aliena Blume, Robinia pseudoaccacia and Pinus densiflora), weed sources: 5 species(Artemisia princeps Pampanini, Erigeron canadensis, Alopecurus aegualis Sobolewski, Echinochloa crusgalli var. frumentacea (Roxb.) Wight and Rumex crispus), and native grass sources: 5 species(Zoysia japonica Steudel, Agropyron tsukushiense var. transiens(Hack) Ohwi., Arundinella hirta(Thunb.) Tanaka, Miscanthus sinensis Anderss and Phragmites comunis Trin). Ten Korean native goats were selected which had nearly the same body weight(average 30.6kg, 남) as experimental animals. Experimental roughages were prepared by 0.5kg per 1 species fed to Korean native goats, and the experiment was carried out from 08:00 to 20:00. The chemical composition and dry matter digestibility of each roughage source and species were significantly different at the sampling area, plant species, growth stages and cutting period (p<0.05). The Korean native goats ate more roughages which had low fibrous contents, but high dry matter digestibility. Among all the 25 species of roughages, the favorite intake species order by Korean native goats was observed like this: Quercus serrata Thunb., Quercus aliena Blume, Trifolium repens, and Pinus densiflora and Artemisia princeps Pampanini which was lower intake compared to other domestic herbivores. On the other hand, compared to each roughage source, total intake amount by Korean native goats was showed as browse(29.9%), legumes(23.0%), weeds(21.6%), grasses(21.5%) and native grasses(4.0%), respectively. Based on the result, the food habit of Korean native goats seems to be closer to browser type.

Outcome of Preimplantation Genetic Diagnosis for Chromosome Aneuploidy and Genetic Disease (유전질환 및 염색체 이상의 예방을 위한 착상전 유전진단의 결과)

  • Kim, Jin-Yeong;Lim, Chun-Kyu;Song, In-Ok;Yoo, Keun-Jai;Yang, Kwang-Moon;Han, Kuk-Sun;Hur, Kuol;Song, Ji-Hong;Jun, Jin-Hyun;Min, Dong-Mi;Park, So-Yeon;Jun, Jong-Young;Koong, Mi-Kyoung;Kang, Inn-Soo
    • Clinical and Experimental Reproductive Medicine
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    • v.29 no.4
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    • pp.269-278
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    • 2002
  • Objective s: Chromosome aneuploidy is associated with recurrent abortion and congenital anomaly and genetic diseases occur repeatedly in the specific families. Preimplantation genetic diagnosis (PGD) can prevent aneuploidy or genetic disease by selecting normal embryos before implantation and is an alternative to prenatal diagnosis. The aim of this study is to assess the outcome of PGD cycles by using FISH or PCR, and to determine the clinical usefulness and values in patients with risk of chromosomal aneuploidy or genetic disease. Materials and Methods: From 1995 to Apr. 2001, a total of 108 PGD cycles in 65 patients with poor reproductive outcome were analyzed. The indications of PGD were translocation (n=49), inversion (n=2), aneuploidy screening (n=7), Duchenne muscular dystrophy (n=5) and spinal muscular atrophy (n=2). PGD was applied due to the history of recurrent abortion, previous birth of affected child or risk of aneuploidy related to sex chromosome aneuploidy or old age. Blastomere biopsy was performed in 6$\sim$10 cell stage embryo after IVF with ICSI. In the single blastomere, chromosome aneuploidy was diagnosed by using FISH and PCR was performed for the diagnosis of exon deletion in DMD or SMA. Results: The FISH or PCR amplification was successful in 94.3% of biopsied blastomeres. The rate of transferable balanced emb ryos was 24.0% in the chromosome translocation and inversion, 57.1% for the DMD and SMA, and 28.8% for the aneuploidy screening. Overall hCG positive rate per transfer was 17.8% (18/101) and clinical pregnancy rate was 13.9% (14/101) (11 term pregnancy, 3 abortion, and 4 biochemical pregnancy). The clinical pregnancy rate of translocation and inversion was 12.9% (11/85) and abortion rate was 27.3% (3/11). In the DMD and SMA, the clinical pregnancy rate was 33.3% (3/9) and all delivered at term. The PGD results were confirmed by amniocentesis and were correct. When the embryos developed to compaction or morula, the pregnancy rate was higher (32%) than that of the cases without compaction (7.2%, p<0.01). Conclusions: PGD by using FISH or PCR is useful to get n ormal pregnancy by reducing spontaneous abortion associated with chromosome aneuploidy in the patients with structural chromosome aberration or risk of aneuploidy and can prevent genetic disease prior to implantation.

Changes of Fatty Acids and Amino Acids Contents of Korean Native Commercial Ducks Meats with Different Raising Periods (사육 기간에 따른 토종 실용오리의 도체 내 지방산과 아미노산 함량 변화)

  • Heo, Kang-Nyeong;Choo, Hyo-Jun;Kim, Chong-Dae;Kim, Sang-Ho;Kim, Hak-Kyu;Lee, Myeong-Ji;Son, Bo-Ram;Choi, Hee-Cheol;Hong, Eui-Chul
    • Korean Journal of Poultry Science
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    • v.40 no.3
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    • pp.235-241
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    • 2013
  • This work was carried to evaluate effect of different raising periods on fatty acids and amino acids properties of Korean native commercial ducks. Korean native ducks (n = 90) from National Institute of Animal Science (RDA, Korea) were divided into 6 groups (15 birds/group) and were fed with meat-type duck diets for 8 wk old. When ducks grew at specific wk (6, 7 and 8 weeks), 2 ducks per group were slaughtered at 6, 7 and 8 wk old. Myristic acid (C14:0) and palmitic acid (C16:0) of saturated fatty acid (SFA) were the highest and arachidonic acid of unsaturated fatty acid (USFA) was the lowest at the age of 6 weeks old (P<0.05) at the age of 6 weeks old among treatments. Stearic acid of SFA was the highest at the age of 7 weeks old among treatments. There was no significant difference on other fatty acids among treatments. SFA and USFA have significant difference at the age of 8 weeks (P<0.05). Histidine, leucine, lysine, methionine, phenylalanine and threonine of essential amino acid increased with passing weeks (P<0.05). There was no significant difference on isoleucine and valine (P>0.05). Every non-essential amino acids were the highest at the age of 8 weeks among treatments (P<0.05). In conclusion, fatty acid contents and amino acid contents increased until the age of 8 weeks old, but further research was needed because of deficiency of data to suggest the correct shipping time.

Present status and prospect for development of mushrooms in Korea

  • Jang, Kab-Yeul;Oh, Youn-Lee;Oh, Minji;Im, Ji-Hoon;Lee, Seul-Ki;Kong, Won-Sik
    • 한국균학회소식:학술대회논문집
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    • 2018.05a
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    • pp.27-27
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    • 2018
  • The production scale of mushroom cultivation in Korea is approximately 600 billion won, which is 1.6% of the Korean gross agricultural output. Annually, ca. 190,000 tons of mushrooms are harvested in Korea. Although the numbers of mushroom farms and cultivators are constantly decreasing, the total mushroom yields are increasing due to the large-scale cultivation facilities and automation. The recent expansion of the well-being trend causes increase in mushroom consumption in Korea: annual per capita consumption of mushroom was 3.9kg ('13) that is a little higher than European's average. Thus the exports of mushrooms, mainly Flammulina velutipes and Pleurotus ostreatus, have been increased since the middle of 2000s. Recently, however, it is slightly reduced. However, Vietnam, Hong Kong, the United States, the Netherlands and continued to export, and the country has increased recently been exported to Australia, Canada, Southeast Asia and so on. Canned foods of Agaricus bisporus was the first exports of the Korean mushroom industry. This business has reached the peak of the sale in 1977-1978. As Korea initiated trade with China in 1980, the international prices of mushrooms were sharply fall that led to shrink the domestic markets. According to the high demand to develop new items to substitute for A. bisporus, oyster mushroom (Pleurotus ostreatus) was received the attention since it seems to suit the taste of Korean consumers. Although log cultivation technique was developed in the early 1970s for oyster mushroom, this method requires a great deal of labor. Thus we developed shelf cultivation technique which is easier to manage and allows the mass production. In this technique, the growing shelf is manly made from fermented rice straw, that is the unique P. ostreatus medium in the world, was used only in South Korea. After then, the use of cotton wastes as an additional material of medium, the productivity. Currently it is developing a standard cultivation techniques and environmental control system that can stably produce mushrooms throughout the year. The increase of oyster mushroom production may activate the domestic market and contribute to the industrial development. In addition, oyster mushroom production technology has a role in forming the basis of the development of bottle cultivation. Developed mushroom cultivation technology using bottles made possible the mass production. In particular, bottle cultivation method using a liquid spawn can be an opportunity to export the F.velutipes and P.eryngii. In addition, the white varieties of F.velutipes were second developed in the world after Japan. We also developed the new A.bisporus cultivar "Sae-ah" that is easy to grown in Korea. To lead the mushroom industry, we will continue to develop the cultivars with an international competitive power and to improve the cultivation techniques. Mushroom research in Korea nowadays focuses on analysis of mushroom genetics in combination with development of new mushroom varieties, mushroom physiology and cultivation. Further studied are environmental factors for cultivation, disease control, development and utilization of mushroom substrate resources, post-harvest management and improvement of marketable traits. Finally, the RDA manages the collection, classification, identification and preservation of mushroom resources. To keep up with the increasing application of biotechnology in agricultural research the genome project of various mushrooms and the draft of the genetic map has just been completed. A broad range of future studies based on this project is anticipated. The mushroom industry in Korea continually grows and its productivity rapidly increases through the development of new mushrooms cultivars and automated plastic bottle cultivation. Consumption of medicinal mushrooms like Ganoderma lucidum and Phellinus linteus is also increasing strongly. Recently, business of edible and medicinal mushrooms was suffering under over-production and problems in distribution. Fortunately, expansion of the mushroom export helped ease the negative effects for the mushroom industry.

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The Pulation Structure of the Pacific Cod (Gadus macrocephalus Tilesius) Based on Mitochondrial DNA Sequences (미토콘드리아 유전자 염기서열 분석에 의한 대구 계군 분석)

  • Seo, Young-Il;Kim, Joo-Il;Oh, Taeg-Yun;Lee, Sun-Kil;Park, Jong-Hwa;Kim, Hee-Yong;Cho, Eun-Seob
    • Journal of Life Science
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    • v.20 no.3
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    • pp.336-344
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    • 2010
  • To assess population structure and genetic diversity among the Pacific cod (Gadus macrocephalus), we investigated mtDNA COI gene sequences of 7 populations. Samples were obtained from Sokcho, Wolsung, Geojedo, Yeosu, Geomundo and Westsouth in 2008 and 2009 (n=28). The sequence analysis of 28 individual samples showed 8 haplotypes, ranging in sequence divergence by pairwise comparisons from 0.2 to 2.2% (1 bp-11 bp). The Gal haplotype was found in Wolsung, Geojedo, Yeosu, Geomundo and Westsouth, and was regarded as the main haplotype of Korean Pacific cod. Ga2, Ga3, Ga6 and Ga7 haplotypes were found only in Sokcho. In the PHYLIP analysis, 8 haplotypes formed two independent groups: cladeA consisted of Ga2, Ga3, Ga6 and Ga7 haplotypes, whereas cladeB contained Gal, Ga4, Ga5 and Ga8 haplotypes. The genetic relationship between the two groups was weakly supported by bootstrap analysis(<50%). In pairwise comparisons between 6 populations other than that from Sokcho, a very high per generation migration ratio ($N_m$=infinite) and a very low level of geographic distance ($F_{sr}=-0.0123-(-0.0423)$) were observed. The estimates of genetic distance between Sokcho and the other localities were all statistically significant (p<0.05, p<0.01, p<0.001), indicating a limited mtDNA-based gene flow between Sokcho and other regions. The finding of the lowest genetic diversity in the Sokcho population (nucleotide diversity=0.00589) may be a result of relatively small population size and interrupted gene flow to other localities. Consequently, the overall considerable migration of Pacific cod population in Korea caused a genetically homogeneous structure to form, although a distinct population was found in this study.

Generation of a Mammalian Gene Expression Vector Using Bovine Viral Diarrhea Virus (Bovine Vira1 Diarrhea Virus를 이용한 포유동물세포 발현벡터의 개발)

  • 이영민
    • Korean Journal of Microbiology
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    • v.38 no.2
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    • pp.86-95
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    • 2002
  • As a result of genome projects, the research to elucidate the function of a protein of interest has recently been well-recognized. In order to facilitate functional genomics, a useful mammalian gene expression vector is required. Using an infectious CDNA clone of BVDV pNADLclns-, we have developed a mammalian gene expression vector. In this study, a replication-competent full-length infectious CDNA clone containing puremycin acetyltransferase (pac) gene (pNADLclns-/pac) was successfully generated. The viral RNA replication and viral protein NS3 synthesis were examined by detecting metabollically $^{32}P$-labelled genomic viral RNA and immunoblotting with a mouse anti-NS3 antibody. To generate viral replicon as an expression vector, we examine if the viral structural genes (C, E0, El, E2) are required for viral replication by deletion analysis. As a result, all of the structural proteins are dispensable for viral replication per se, but essential for infectious viral particle formation. Based on our deletion analysis, we have generated a replication-competent BVDV viral replicon (pNADLclns-/pac/${\Delta}S$), whose structural genes are all deleted. In addition to NADLclns- /pac/${\Delta}S$, NADLclns-/ luc/${\Delta}S$ viral replicon containing luciferase gene as a reporter was constructed and fecund to be replication-compotent in HeLa and BHK cells as well as MDBK cells. Therefore, BVDV viral replicon developed in our study will be a useful tool to express a protein of interest in various mammalian cells.

Comparison of Pakistani Ephedra Herba and Chinese Ephedra Herba Containing Gangjihwan in the Improvement Effects of Weight Loss in a High Fat Diet-Fed Obese Mice (고지방식이 비만마우스 모델에서 파키스탄산 및 중국산 마황으로 조성된 강지환(降脂丸)의 체중감량 효과 비교)

  • Kim, Byeong Chul;Seok, Hoa Jun;Yoo, Jae Sang;Ku, Ja Ryong;Yoon, Ki Hyeon;Jo, Ju Heum;Jang, Du Hyon;Jung, Yang Sam;Kim, Jong Hoon;Ahn, Ye Ji;Woo, Sangee;Yoon, Miso;Shin, Soon Shik
    • Herbal Formula Science
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    • v.22 no.2
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    • pp.63-76
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    • 2014
  • Objectives : This study investigated the improvement effects of Pakistani (DF-a) and Chinese Ephedra herba-containing Gangjihwan (DF-b) on obesity in a high fat diet-fed obese mouse model. Methods : Eight-week-old C57BL/6N mice were divided into four groups: a normal lean group given a standard diet, an obese control group given a high fat diet, and DF-a and DF-b groups given a high fat diet with DF-a (80 mg/kg), and DF-b (80 mg/kg), respectively. After 8 weeks of treatment, body weight gain, feeding efficiency ratio, blood lipid markers, fat weight and histology were examined. Results : 1. Body weight gain and fat mass were significantly decreased in DF-a and DF-b groups compared with control. The extent of decreases was eminent in DF-a group. 2. Feeding efficiency ratio and circulating leptin concentration were significantly decreased in DF-a and DF-b groups compared with control, whereas circulating adiponectin concentration was increased in DF-a and DF-b groups compared with control. 3. Consistent with their effects on body weight gain and fat mass, circulating concentrations of triglyceride, glucose and insulin were decreased in DF-a and DF-b groups compared with control. 4. The size of adipocytes were decreased by DF-a and DF-b compared with control, whereas the adipocyte number per unit area was increased by them, suggesting that DF-a and DF-b decreased the number of large adipocytes. 5. Consistent with their effects on body weight gain, liver fibrosis was reduced in DF-a and DF-b groups compared with control. Conclusions : In conclusion, these results suggest that DF-a and DF-b not only decrease feeding efficiency ratio, plasma leptin concentration, and blood anti-obesity biomarkers, but also reduce fat mass, contributing to the improvement of obesity. DF-a and DF-b also inhibit liver fibrosis. In addition, these effects were similar between Pakistani Ephedra herba and Chinese Ephedra herba-containing Gangjihwan.