• Title/Summary/Keyword: PCR

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An Ecological Analysis of Lichens Distributed in Rocks of Coast and Field in U-do by Molecular Technique (분자생물학적 기법에 의한 우도해안과 노지암석에 분포하는 지의류의 생태학적 분석)

  • Gang, Hyeong-Il;Yun, Byeong-Jun;Kim, Seong-Hyeon;Sin, Deok-Ja;Kim, Hyeon-U;Heo, Jae-Seon;Gang, Ui-Seong;O, Gye-Heon;Go, Yeong-Jin
    • Microbiology and Biotechnology Letters
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    • v.32 no.4
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    • pp.334-340
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    • 2004
  • This study was initially performed to obtain fundamental data on lichen distribution depending on environmental condiLichens distributed on coastal rocks atleast 20 m far away from the coast were also investigated. Analyof morphological characteristics and rDNA ITS clones revealed that lichens containing 9 families and 15 genera were distributed in coastal rocks of V-do, while lichens containing 10 families and 14 genera were in field rocks, demonstrating little difference of diversity between both sites. Foliose lichens such as PhaeophysPhyscia, Pyxine belonging to the family Physciaceae were representatives distributed in coastal rocks of V-do, along with Xanthoparmelia belonging to Parmeliaceae, fruticose lichens such as Ramalina to Ramaliand crustose lichen Lecanora to Lecanoraceae. In contrast, foliose lichen Cladonia belonging to the family Cladoniaceae, fruitcose lichen Stereocaulon to Streocaulaceae, and crustose lichen Porpidia to Porpidwere found only on the surface of field rocks. Crustose lichens containing genera Caloplaca, CanDirinaria, Graphis, Rhizocarpon, and Pertusaria were rarely distributed in coastal rocks, and most of them were also found in field rocks.

Effects of replacing fish oil with palm oil in diets of Nile tilapia (Oreochromis niloticus) on muscle biochemical composition, enzyme activities, and mRNA expression of growth-related genes

  • Ayisi, Christian Larbi;Zhao, Jinliang;Yame, Chen;Apraku, Andrews;Debra, Grace
    • Fisheries and Aquatic Sciences
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    • v.22 no.11
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    • pp.25.1-25.9
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    • 2019
  • Background: Due to the continuous demand for fish coupled with decline in capture fisheries, there is the need to increase aquaculture production to meet the demand. Aquaculture is faced with high cost of feeding since fish oil and fish meal are expensive. In view of this, there are calls to explore alternatives that are cheap and reliable. Objectives: This study on Oreochromis niloticus was conducted to evaluate the effects of replacing fish oil (FO) with palm oil (PO) at 0%, 25%, 50%, 75%, and 100% on muscle fatty acid and proximate composition as well as growthrelated enzyme activities and mRNA expression. Methods: Oreochromis niloticus were fed five experimental diets (33% crude protein and 10% crude lipid) for 8 weeks. Feed had variation in fish oil and palm oil contents. After the 8 weeks feeding trial, five fish were sampled from each tank (15 from each treatment) and euthanized using an excess dose of tricaine methane sulfonate (MS-222 at 200 mg/L). Fatty acid and enzyme activities were analyzed using standard protocols. Also, RT-qPCR was used to quantify the expression levels of selected growth-related genes. Results: Fish fed 25% PO recorded the least muscle protein content and was significantly lower than the group fed 100% PO. Paired box protein 7 (Pax-7) enzyme activity was significantly higher in the group fed 50% PO compared to the groups fed 25% PO and 100% PO, while caplain-3 (Capn-3) was significantly lower in the group fed 0% PO compared to all other groups. There was a significant difference among treatments with respect to mRNA expression of Pax-7 and Capn-3. Group fed 25% PO had significantly lower mRNA expression of Pax-7, while the group fed 75% PO recorded significantly higher mRNA expression of Capn-3 compared to groups fed 0% PO, 25% PO, and 100% PO. Pearson's correlation analysis revealed that Igf-I and Igf-II mRNA expression have significant correlation with n-3 polyunsaturated fatty acids content in muscle. Conclusion: The results suggest muscle protein content could be modified if FO is replaced with PO. Also, mRNA expression of Pax-7 and Capn-3 is affected by replacing FO with PO.

Occurrence of Viruses Infecting Foxtail Millet (Setaria italica) in South Korea (국내에 발생하는 조 바이러스의 종류 및 발생 실태)

  • Park, Chung Youl;Min, Hyun-Geun;Lee, Hong-Kyu;Yeom, Yoon Ah;Oh, Jonghee;Kim, Bong-Sub;Bae, Dae-Hyeon;Yoon, Young-Nam;Lee, Su-Heon
    • Research in Plant Disease
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    • v.23 no.1
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    • pp.69-74
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    • 2017
  • In 2015, a nationwide survey was carried out to investigate about occurrence pattern of virus infecting foxtail millet. A total 100 foxtail millet leaf samples showing virus-like and abnormal symptoms were collected in the seven main cultivated regions of Korea. Four viruses were identified using reverse transcription polymerase chain reaction and RNA sequencing. Of the collected 100 foxtail millet samples, 10 were Barley virus G (BVG), 4 were Rice stripe virus (RSV), 1 was Northern cereal mosaic virus (NCMV), and 1 was Sugarcane yellow leaf virus (ScYLV) infection. To our best knowledge, this is the first report of BVG and NCMV infecting foxtail millet in Korea and ScYLV is expected as new Polerovirus species. This research will be useful in breeding for improved disease-resistant foxtail millet cultivars.

Prevalence of Canine Coronaviral Enteritis in Korea (개 코로나바이러스성 장염의 발생 역학)

  • Jeoung, Seok-Young;Ahn, So-Jeo;Pak, Son-Il;Kim, Doo
    • Journal of Veterinary Clinics
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    • v.27 no.3
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    • pp.209-215
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    • 2010
  • Canine coronavirus (CCV) is a cause of sporadic outbreaks of enteritis in dogs. This study was performed to carry out epidemiological investigation on the recent outbreaks of CCV enteritis of dogs and determined the potential prognostic factors affecting the survival of dogs. The 131 (34.4%) out of 381 fecal samples collected from dogs with enteritis were positive for CCV by RT-PCR. The fecal samples contained genotype I (30.5%), genotype II (29.0%), and both genotypes (40.5%) of CCV. The majority of dogs with CCV infection ranged 6-18 weeks of age. Age over 18 weeks was significantly associated with higher survival rate (P < 0.05). Of the clinical signs examined, dogs without anorexia were significantly higher survival rate (P < 0.01). The 90.1% of dogs with CCV infection were co-infected by CCV and CPV-2. From the results of this study, it can be concluded that CCV infection is widespread in the Korean dog population and CCV may be attributed to be one of the important agents causing enteritis in pups.

Induction of Apoptosis by Curcuma aromatica on Lung Cancer Cells(A549), Cervical Cancer Cells(HeLa), Glioma Cancer Cells(A172) and Prostate Cancer Cells(PC3) (울금(鬱金)이 폐암(肺癌), 자궁암(子宮癌), 신경교종(神經膠腫) 및 전립선암(前立腺癌)에 대한 세포자살유도(細胞自殺誘導)에 미치는 영향(影響))

  • Park, Sang-Hyun;Kim, Jin-Sung;Yoon, Sang-Hyub;Ryu, Bong-Ha
    • The Journal of Internal Korean Medicine
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    • v.27 no.2
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    • pp.379-393
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    • 2006
  • Objectives: We are aimed to identify anti-tumor effects of Curcuma aromatics on some kinds of cancer cells through molecular biologic methods. Materials & Methods: We used 4 kinds of cancer cell lines such as lung cancer cells(AS49), cervical cancer cells(HeLa), glioma cancer cells(A172) and prostate cancer cells(PC3). We treated the boiled extract of Curcuma aromatica $5{\mu}g,\;10{\mu}g$ to cultural media(ml) for 24 hours. We measured the cytotoxicitv on 4 kinds of cancer cells through tryphan blue exclusion test and the suppressive effect on viability of 4 kinds of cancer cells via MTT assay. We measured change of mitochondria membrane potential via flow cytometry. The quantitative RT-PCR was used to examine the effect on the revelation of Bcl-2 and Bax which are genes related to apoptosis. We examined the effect on the revelation of Bcl-2 Protein and Bar protein by western blot analysis. Results : In the experiment of tryphan blue exclusion test, the extract of Curcuma aromatica showed more significant killing effect on AS49, HeLa than the control group with density dependent manner, which was statistically significant. In the experiment of MTT assay the extract of Curcuma aromatica showed more suppressive effect on viability of A549, HeLa than the control group with density dependent manner, which was statistically significant. Curcuma aromatica induced apoptosis by decreasing the membrane potential of mitochondria in A549, HeLa. In the experiment of the revelation of genes related to apoptosis, the revelation of Bcl-2 decreased and the revelation of Bax increased in A549, HeLa treated with Curcuma aromatica with dose dependent manner. In the experiment of the revelation of protein related to apoptosis, the protein levels of Bcl-2 decreased and the protein levels of Bax increased in AS49, HeLa treated with Curcuma aromatica with dose dependent manner. Conclusions: From this study, we can infer that Curcuma aromatica has anti-tumor effect on lung cancer cells and uterine carcinoma cells but not on glioma cells and prostate cancer cells.

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Effects of Curcuma longa L. on Some Kinds of Cancer Cells (강황이 수종의 암세포에 미치는 영향)

  • Yoon, Joo-Ho;Kim, Jin-Sung;Yoon, Sang-Hyub;Ryu, Bong-Ha
    • The Journal of Internal Korean Medicine
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    • v.27 no.2
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    • pp.429-443
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    • 2006
  • Objectives : The Purpose of this study was to identify anti-tumor effects of Curcuma longa L. on some kinds of cancer cells through molecular biologic methods. Materials & Methods : We used 4 kinds of cancer cell lines such as glioma cells(A172), cervical cancer cells(HeLa), Prostate cancer cells(PC3), lung cancer cells(A549). We injected the boiled extract of Curcuma longa L. $5{\mu}g,\;10{\mu}g$ to culture media(ml) for 24 hours. We measured the cytotoxic effect on 4 kinds of cancer cells through trypan blue exclusion test and the suppressive effect on viability of 4 kinds of cancer cells via MTT assay. We measured the change of mitochondria membrane potential via flow cytometry. The quantitative RT-PCR was used to examine the effect on the revelation of Bcl-2 and Bax which genes are related to apoptosis. We examined the effect on the revelation of Bcl-2 protein and Bax protein by western blot analysis. Results: 1. Extract of Curcuma longa L. showed significant cytotoxic effect on A172, HeLa, PC3 compared to the control group with density dependent manner. 2. Extract of Curcuma longs L. showed significant suppressive effect on viability of A172, HeLa, PC3 compared to the control group with density dependent manner. 3. Curcuma longs L. induced apoptosis by decreasing the membrane potential of mitochondria in A172, HeLa, PC3. 4. In the test about the revelation of genes related to apoptosis, the revelation of Bcl-2 decreased and the revelation of Bax increased in A172. HeLa, PC3 treated with Curcuma longa L. with density dependent manner. 5. In the test about the revelation of protein related to apoptosis, the protein levels of Bcl-2 decreased and the protein levels of Bax increased in A172, HeLa, PC3 treated with Curcuma longa L. Conclusions: This experiment shewed that Curcuma longs L. has anti-tumor effect on glioma, cervical, Prostate cancer cells except on lung cancer. We hope that anti-tumor effects of Curcuma longa L. will be more Practically identified.

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The Inhibitory Effects of Sabaek-san and Sabaeksan plus Sasam on the IL-6, IL-8 and GM-CSF mRNA Levels in Human Epithelial Cells (사백산(瀉白散)과 사백산가사삼(瀉白散加沙蔘)이 인간 기관지상피세포의 IL-6, IL-8 및 GM-CSF 발현에 미치는 영향)

  • Jung, Hee-Jae;Jung, Sung-Ki;Rhee, Hyung-Koo;Chung, Kwang-Jin
    • The Journal of Internal Korean Medicine
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    • v.23 no.1
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    • pp.15-23
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    • 2002
  • Objective : We aimed to identify the dose-dependent inhibitory effects of Sabaek-san(瀉白散) and Sabaeksan plus Sasam(Adenophorae Radix) 瀉白散加沙蔘) on the mRNA expressions of Interleukin(IL)-6, IL-8 and granulocyte macrophage colony stimulating factor(GM-CSF) involved in the asthma model. Materials and Methods : Through this study, BEAS-2B cell lines, human epithelial cells were used. These cells were stimulated by tumor necrosis factor(TNF)-${\alpha}$, IL-1${\beta}$ and histamine for artificial inflammatory expression. ${\beta}$-action messenger RNA(mRNA) was used for standards. After each 24hours of Sabaeksan and Sabaeksan plus Sasam treatment, total cellular RNAs were collected by treating RNA zol directly on living cells, Then the transcriptional activities of IL-6, 8 and GM-CSF were measured by RT-PCR with electrophoresis, Results : The mRNA expressions of IL-6 are significantly inhibited compared to those of controlled group at 40 and 100ug/ml of Sabaeksan extract and $100{\mu}g/ml$ of Sabaeksan plus Sasam extract (p<0.05). The mRNA expressions of IL-8 are significantly inhibited compared to that of controlled group at 2.40 and 100 ug/ml of Sabaeksan extract and $40.100{\mu}g/ml$ of Sabaeksan plus Sasam extract(p<0.05) THe mRNA expressions of GM-CSF are significantly inhibited compared to those of the controlled group at $100{\mu}g/ml$ of Sabaeksan extract adn $40.100{\mu}g/ml$ of Sabaeksan plus Sasam extract.(p<0.05) Conclusions : This study shows that Sabaeksan and Sabaeksan plus Sasam have dose-dependent inhibitory effects on the mRNA expressions of IL-6, IL-8 and GM-CSF in human epithelial cells. Therefore, these types of herb medicine may inhibit the inflammatory process of asthma. Advanced studies are required to investigate the mechanisms of inhibition by herb medicine in the asthma model.

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The Association of the -675 4G/5G and A-844G Polymorphisms of the Plasminogen Activator Inhibitor-1 Gene with a Risk of Ischemic Stroke in Korean Population (한국인의 허혈성 뇌경색 발생과 플라스미노겐 활성 억제인자-1(PAI-1) 유전자 다형성과의 관계)

  • Lee, Byung-Cheol;Byun, Sang-Hyuk;Kim, Soon-Il;Kang, Ki-Hoon;Ahn, Se-Young;Doo, Ho-Kyung;Seo, Jung-Chul;Leem, Kang-Hyun;Cho, Seong-Ho;Oh, Chad K.;Ahn, Young-Min
    • The Journal of Internal Korean Medicine
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    • v.25 no.4
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    • pp.18-24
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    • 2004
  • 연구배경 : 플라스미노겐 활성 억제인자-1 (plasminogen activator inhibitor-1; PAI-1)은 허혈성 뇌경색의 발생의 원인이 되는 섬유소 용해작용의 저하를 매개하는 인자로서, PAI-1의 작용이 촉진되면 섬유소 용해기능이 저하되어 관상동맥 및 뇌혈관질환의 발생을 증가시키게 된다. PAI-1 유전자의 촉진자(promoter) 영역에는 -675 4G/5G (4G/5G)와A -844G (A/G)의 두 개의 유전자 다형성이 존재하며, 이는 PAI-1의 유전자 전사과정에 영향을 미쳐 혈청 PAI-1의 농도를 증가시키고 결과적으로 허혈성 뇌경색의 발생확률을 높이는 작용을 하게 된다. 연구방법 : 허혈성 뇌경색으로 진단 받은 167명의 환자와 173명의 건강인의 말초혈액에서 DNA를 분리한 후 PAI-1의 4G/5G와 A/G 유전자 다형성에 대한 연쇄중합반응 및 제한효소 절편길이 다형성 (polymerase chain reaction-restriction fragment length polymorphism; PCR-RFLP) 방법을 이용하여 허혈성 뇌경색 발생과 유전자 다형성과의 관계를 비교 분석하였다. 결과 : 허혈성 뇌경색 환자에서의 4G/4G의 유전자형의 빈도는 15.0%으로 정상 대조군의 33.5%에 비해 현저하게 낮게 나타났다 (P < 0.0001). 각각의 유전자형과 허혈성 뇌경색의 발생 위험도 (odd ratio ; OR)와의 관계를 분석했을 때 4G/4G 유전자형을 가질 경우 위험도는 0.35배로 현저하게 낮아졌으며, (P < 0.0001), 5G/5G 유전자형을 가질 경우 위험도는 4.49배 로 현저하게 높아졌다 (P < 0.0001). 그러나, A/G 유전자 다형성과 허혈성 뇌경색의 발생과는 유의한 연관성을 발견하지 못하였다. 결론 : 이상의 결과로 볼 때 PAI-1 유전자의 4G/4G 유전자형은 허혈성 뇌경색의 발생 비율을 감소시키는 작용을 하는 것으로 여겨진다.

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A Study the Effects of Three Preparations of Hirudo on the Expression of Pro-inflammatory Cytokines in Human Bronchial Epithelial Cells Line BEAS-2B (수질(水蛭)의 제법(制法)에 따른 BEAS-2B 인간(人間) 기관지상피세포(氣管支上皮細胞)의 염증유발성(炎症誘發性) Cytokines 발현(發顯)에 미치는 영향(影響))

  • Jung, Hee-Jae;Jung, Sung-Ki;Rhee, Hyung-Koo;Han, Dong-Ha
    • The Journal of Internal Korean Medicine
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    • v.25 no.4
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    • pp.260-273
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    • 2004
  • Backgrounds : In recent years, asthma has become recognized as a chronic inflammatory disease associated pathologically with airway epithelial inflammation and airway remodeling. Objectives : To evaluate the different effects of Hirudo depending upon pharmaceutical manufactures on the expression and the activities of IL-6 and GM-CSF in airway epithelial cells, samples of Hirudo(水蛭), Hirudo toasted with Talcum(水蛭滑石炒) and Hirudo toasted with Ephedrae Herba(水蛭麻黃炒) were tested. Methods : After inducing enhanced messenger RNA(mRNA) expression and secretion of each cytokine by tumor necrosis factor-alpha(10 ng/ml) treatment, cultured human bronchial epithelial cell line BEAS-2B was added to each sample$(l,\;10,\;100\;&\;1000\;{\mu}g/ml)$. Subsequently, DNA activities were analyzed. Specifically mRNA expression and culture supernatants(protein levels) of IL-6 and GM-CSF from BEAS-2B cells, were analyzed using luciferase reporter gene assay, reverse transcription-polymerase chain reaction(RT-PCR) analysis and enzyme-linked immunosorbent assay. Results : Hirudo toasted with Ephedrae Herba(水蛭麻黃炒) and Hirudo(水蛭) inhibited IL-6 activities in BEAS-2B cells remarkably, and inhibited mRNA expression levels and protein levels in supernatant of IL-6 and GM-CSF at various concentrations, significantly(p<0.05). However, Hirudo toasted with Talcum(水蛭滑石炒) had no effect on mRNA expression levels and showed a slight inhibitory effect on GM-CSF protein levels in supernatant of culture medium. Conclusions : These results strongly suggest that Hirudo toasted with Ephedrae Herba(水蛭麻黃炒) and Hirudo(水蛭) would be serve as effective medicaments in the treatment of airway inflammation and remodeling of asthmatic patients.

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Inhibitory Effect of YBR on Hepatic Fibrogenesis (YBR의 간섬유화(肝纖維化)억제 효과(效果)에 관한 연구(硏究))

  • Seung, Hyun-Seok;Woo, Hong-Jung
    • The Journal of Internal Korean Medicine
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    • v.31 no.2
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    • pp.314-330
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    • 2010
  • Objective : This study was performed to investigate the anti-fibrogenic effect and changes of inflammation-related genes by YBR I and YBR II (YBR I: Arteisiae Capillaris Herba, Atractylodis Rhizoma Alba, Hoelen/ YBR II: YBR I +Sanguisorbae Radix, Biotae Cacumen, Cirsii Japonici Herba) on HSC(hepatic stellate cells)-T6 and TAA-induced rat liver tissue. Materials and Methods : HSC-T6 were treated with various concentrations of distilled-water extract YBR I and YBR II extract for 24, 48 and 72 hours. After the treatment, cell viability, proliferation, procollagen levels and IL-6 levels were measured by using MTT Assay, BrdU Assay, Procollagen Type 1 C-peptide EIA kit, and Murine IL-6 ELISA Development kit. Rat liver fibrosis was induced by intraperitoneal TAA injection of 150mg/kg 3 times a week for 6 weeks. After the treatment, body weight, liver & spleen weights, liver function test, complete blood cell count and change of portal pressure were studied. In addition, gene expressions of ASMA, IL-6, MMP-2, TIMP-1 and TIMP-2, all of which are known to be associated with liver fibrosis, were analyzed by using Real-Time PCR. After YBR I and YBR IItreatment, percentages of collagen in TAA-induced rat liver tissue were measured. Results : The viability and proliferation of the HSC-T6 decreased as the concentration increased. The production of procollagen decreased as the concentration increased. The production of IL-6 was little influenced by YBR I and YBR II. There was no difference in rat body weight between the TAA-only group and the YBR groups. Compared with rat liver weight of TAA-only group, that of the YBR groups increased. In the YBR I group, the serum level of AST elevated by TAA injection significantly decreased and in the YBR I and II group, the serum level of ALP and ALT elevated by TAA injection decreased. In the YBR I group, white blood cell count elevated by TAA injection decreased but platelets increased. In the YBR I group, the portal pressure elevated by TAA injection significantly decreased. Decreases in the gene expression of ASMA and MMP-2 were observed in the YBR I group. The gene expression of IL-6 was little influenced by YBR I and YBR II -treated groups. In the histological finding, TAA injections caused severe fibrosis, but YBR I and YBR II treatment significantly reduced the amounts of hepatic collagens. Conclusions : These results suggest that YBR I and II have inhibitory effects on the hepatic fibrogenesis.