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The Effects of Herba Chelidonii Extracts on Calcitonin Gene-Related Peptide and Substance P Immunoreactive Response in Spinal Cord and Ganglia of Adjuvant-Induced Arthritis (관절염 모델에서 백굴채전탕액이 척수와 척수신경절의 Calcitonin Gene-Related Peptide와 Substance P 면역반응에 미치는 영향)

  • Park Jong Joo;Yook Tae Han;Song Beem Yong;Lee Kwang Gyu;Yu Yun Jo;Lee Chang Hyun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.2
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    • pp.272-278
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    • 2002
  • To investigate the anti-inflammatory and analgesic effects of Herba Chelidoniie, the extracts of Herba Chelidoniie treated in arthritic rat model. Complete Freund,s Adjuvant(CFA) were injected in the subcutaneous tissue of left foot paw of rats to induce arthritis. Herba Chelidonii extracts(HC) was administered immediately into the peritoneal cavity after CFA injection for 12 days. The immunohistochemical stainings for calcitonin gene-related peptide(CGRP) and substance P in the L4, L5 and L6 spinal dorsal horn and ganglia were done, and the paw swelling was measured with a micrometer and the blood leukocytes were counted. The results were as follows : The paw swelling of HC treated group was significantly decreased in 12th day after CFA injection compare to control group. The change of differential leukocytes counts of HC treated group increased the ratio of lymphocytes, and decreased the ratio of neutrophils compare to control group. The extent of CGRP immunoreactive nerve fiber of dorsal horn of HC treated group was weakly stained compare to control group. The number of CGRP immunoreactive neurons of L6 spinal cord of HC treated group was significantly decreased compare to control group. The extent of substance P immunoreactive nerve fiber of dorsal horn of He treated group was weakly stained compare to control group. The number of substance P immunoreactive neurons of L4, L5 and L6 spinal cord of HC treated group was significantly decreased compare to control group. These experimental results suggest that Herba Chelidonii extracts reduce the number of CGRP and substance P immunoreactive neurons and nerve fibers of spinal dorsal horns and ganglia, and decrease paw swelling in arthritic rat model, which may be closely related to analgesic and antiinflammatory effects of Herba Chelidonii.

Experimental studies on antitumor effects and immune responses of Eunwhasagantang and Eunwhasagantangganokyong (은화사간탕(銀花瀉肝湯)과 은화사간탕가녹용(銀花瀉肝湯加鹿茸)의 항암효과(抗癌效果)와 면역반응(免疫反應)에 관(關)한 연구(硏究))

  • Kim, Jin-Seong;Ryu, Bong-Ha;Park, Dong-Won;Ryu, Gi-Won
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.3 no.1
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    • pp.1-27
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    • 1997
  • This study was performed to investigate the effects of Eunwhasagantang and Eunwhasagantangganokyong on the viability of tumor cells in vitro(MTT assay), on antitumor effects after Sarcoma-180 cells transplantation into the peritoneal cavity or left groin, and on decreased immune responses in mice induced by methotrexate. The extracts of its herbal medicines were orally administered for 14 or 21 days. To evaluate the effects of the Eunwhasagantang and Eunwhasagantangganokyong many items such as 50% inhibitory concentration($IC_{50}$), mean survival days, tumor and body weight for antitumor effects, and delayed type hypersensitivity, hemagglutinin titer, hemolysin titer, rosette forming cells, natural killer cell activity, lymphocyte transformation, productivity of interleukin-2 and phagocytic activity for immune responses were measured in ICR mice. The results were obtained as follows; 1. $IC_{50}$ of Eunwhasagantang treated group was 0.000204mg/ml on SNU-396 and that of Eunwhasagantangganokyong treated group was 0.000103mg/ml on SNU-1, those results indicate that the medicine has high antitumor activity. 2. Mean survival times in Eunwhasagantang and Eunwhasagantangganokyong treated groups were slightly increased with no significance, as compared with the control group. 3. Tumor weight in Eunwhasagantang and Eunwhasagantangganokyong treated group was depressed, as compared with the control group(p<0.01). 4. Body weight in Eunwhasagantang and Eunwhasagantangganokyong treated group was significantly increased, as compared with the control group(p<0.05). 5. Delayed type hypersensitivity in Eunwhasagantang and Eunwhasagantang-ganokyong treated group was slightly decreased with no significance, as compared with the control group. 6. Hemagglutinin titer in Eunwhasagantang and Eunwhasagantangganokyong treated group was slightly increased with no significance, as compared with the control group. 7. Hemolysin titer only in Eunwhasagantang treated group was significantly increased, as compared with the control group(p<0.01). 8. Rosette forming cells only in Eunwhasagantangganokyong treated group was slightly increased with no significance, as compared with the control group. 9. In the NK cell activity, the ratio of effector cells and target cells of the Eunwhasagantang treated group was significantly increased(p<0.01) in case which the ratio was 100: 1, and that of the Eunwhasagantangganokyong treated group was significantly increased(P<.01, p<0.05) in case which the ratio was 100:1, 50:1, as compared with the control group. 10. Lymphocyte trasnformation in Eunwhasagantang and Eunwhasagantangganokyong treated group was significantly increased, as compared with the control group(p<0.01). 11. Interleukin-2 in Eunwhasagantang and Eunwhasagantangganokyong treated group was significantly increased, as compared with the control group(p<0.05, p<0.01). 12. Phagocytic activity in Eunwhasagantang and Eunwhasagantangganokyong treated group was significantly increased, as compared with the control group(p<0.05). According to the above results, it could be suggested that Eunwhasagantang and Eunwhasagantangganokyong have prominent antitumor effects, and enhance both cellular and humoral immunity.

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Experimental studies on antitumor effects and immune responses of Kyegyoksan (계격산(啓膈散)의 항암(抗癌) 및 면역반응(免疫反應)에 관(關)한 실험적(實驗的) 연구(硏究))

  • Lee, Ji-Hyang;Ryu, Bong-Ha;Park, Dong-Won;Ryu, Gi-Won
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.3 no.1
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    • pp.99-128
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    • 1997
  • In order to investigate the effects of Kyegyoksan on antitumor effects after Sarcoma 180 cells transplantation into the peritoneal cavity or left groin in mice, and immune depression in mice induced by methotrexate, the extracts of its herbal medicines were orally administered for 14 or 21 days. Experimental studies were performed for measureance of $IC_{50}$ in MTT assay, mean survival days, tumor and body weights for antitumor effects, delayed type hypersensitivity, hemagglutinine titer, hemolysin titer, rosette forming cells, interleukin-2 productivity, lymphocyte transformation, natural killer cell activity and phagocytic activity for immune responses in the immune depressed ICR mice, and SGOT, SGPT, BUN and creatinine for liver and kidney protective function in SO-rats. The results were obtained as follows: 1. From the results of MTT assay, the Kyegyoksan exstracts for SUN-1 and SUN-C4 were inhibited cell viability. 2. Mean survival time in Kyegyoksan-treated group was slightly increased with no effectiveness, as compared with the control group. 3. Tumor weight in Kyegyoksan-treated group was depressed with the statistical significance, as compared with the control group(p<0.01). 4. Body weight in Kyegyoksan-treated group was incresed with the statistical significance, as compared with the control group(p<0.05). 5. Delayed type hypersensitivity in Kyegyoksan-treated group was slightly incresed with no effctiveness, as compared with the control group. 6. Hemagglutinin titer in Kyegyoksan-treated group was incresed with the statistical significance(p<0.05), but hemolysin titer was slightly incresed with no effectiveness, as compared with the control group. 7. Rosette forming cells in Kyegyoksan-treated group was incresed with the statistical significance, as compared with the control group(p<0.001). 8. Interleukin-2 productivity in Kyegyoksan-treated group was incresed with the statistical significance, as compared with the control group(p<0.001). 9. Lymphocyte transfomation in Kyegyoksan-treated group was incresed with the statistical significance, as compared with the control group(p<0.01). 10. Natural killer cell activity in Kyegyoksan-treated group at E/T ratio 100 : 1 was incresed with the statistical significance(p<0.01), but at E/T ratio 50 : 1 and 10 : 1 was slightly incresed with no effectiveness, as compared with the control group. 11. Phagocytic activity in Kyegyoksan-treated group was slightly incresed with no effectiveness, as compared with the control group. 12. The levels of serum glutamic oxalacetic transaminase, serum glutamic pyruvic transaminase, blood urea nitrogen and serum creatinine in Kyegyoksan-treated group were not effective change, as compared with the control group. According to the above results, it could be suggested that Kyegyoksan have prominent antitumor effects, enhance both cellular and humoral immunity, and have no injury to liver and kidney functions.

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A Study on Superoxide Dismutase from various Tissue of the Tricarboxylic acid cycle blocked Rat (Tricarboxylic acid회로를 차단한 흰쥐의 조직에서 Superoxide Dismutase에 관한 연구)

  • Kim, Yil
    • Korean Journal of Microbiology
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    • v.23 no.1
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    • pp.69-76
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    • 1985
  • This study was carried out to observe the formation of superoxide radicals and the changes in the activities of superoxide dismutase (EC.1.15.1.1.) from the various organs of a rat which was blocked tricarboxylic acid cycle. In order to block the tricarboxylic acid cycle, the beta-fluoroethylacetate was injected into peritoneal cavity of rat and removed the various tissues from the rat at internals of an hour. By tissue extracts being prepared by the method of Weigiger and Fridovich the activities of superoxide dismutase, aconitase, and contents of bliid glucose, citrates, and wuperoxide radicals were determined. The experimental results are summarized as follows: Accumulation of citrates if increased within three hours after treatment in the all tested tissues, especially, in the geart and spleen they are higher than one of other tissues as 12 and 20 times of control. The activities of aconitase are ingibited to 30-35% on an hour after beta-fluoroethylacetate treatment comparing with that of control rat. The content of blood glucose is increased to 1.6 fold of normal value after 5 hours of treatment. In all tested tissues, superoxide radicals are formed in the heart as 0.26 micromoles per gram tissue between one and three hours after treatment. The activities of total superoxide dismutase are increased between one and three hours after treatment in the all tested tissues and one of these enzymes in heart is highest. The activities of superoxide dismutase containing Mn are also increased with an increase of total superoxide dismutase activities.

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The Current Research Methodology of Pharmacopucture for the Treatment of Animal Cancer Models in Korea (암에 대한 약침치료의 국내 동물모델 연구 현황)

  • Ryu, Hee Kyoung;Goo, Bon Hyuk;Suk, Kyung Hwan;Lee, Ju Hyeon;Ryu, Soo Hyeong;Lee, Su Yeon;Kim, Min Jeong;Park, Yeon Cheol;Baek, Yong Hyeon;Park, Dong Suk;Seo, Byung Kwan
    • Journal of Acupuncture Research
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    • v.31 no.4
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    • pp.81-97
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    • 2014
  • Objectives : The purpose of this study is analyzing the current research methodology of pharmacopucture for the treatment of animal cancer models. Methods : Four electronic databases were searched for animal studies published from January 2000 to September 2014 onward using these search terms "cancer, anticancer, pharmacopuncture, beevenom". Selected articles were described about animal cancer models. The methods used to induce cancer and the outcome measures used to assess the effects of pharmacopuncture on animal cancer models were analyzed. Results : 37 articles were included. For producing animal cancer models BALB/C mice(n=22) and C57BL/6 mice(n=17) were selected. And intravenous injection of B16-F10 melanoma cells into tail vein(n=14) or intraperitoneal injection of sarcoma-180 cells(n=14) were frequently used to induce cancer. Various pharmacopunctures were injected into acupoints $CV_{12}(n=19)$, $ST_{36}(n=8)$, $BL_{18}(n=8)$ or peritoneal cavity(n=6), tumor site(n=2), tail vein(n=2). Outcome measures were categorized into anti-cancer, anti-metastasis, general condition, cytotoxicity, immune response, toxicity. Median Survival Time(MST) and increase of life span(ILS)(n=26) was frequently used for evaluating anti-cancer effects. And pulmonary colonization assay(n=13) was frequently used for evaluating anti-metastasis effects Conclusions : Based on these data, further research would be needed to ascertain the effectiveness of pharmacopuncture for treating cancer and broaden the range of clinical applications.

Prevalent Characteristics of Fowl Typhoid and Antibiotic Susceptibility of Salmonella gallinarum (닭 티푸스의 발생상황과 Salmonella gallinarum의 항균제 감수성)

  • 박노찬;도재철;조광현;장성준;권헌일;박덕상
    • Korean Journal of Veterinary Service
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    • v.18 no.2
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    • pp.113-123
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    • 1995
  • The present study was conducted to investigate the prevalent characteristics of Fowl Typhoid and antibiotic susceptibility of Salmonella gallinarum isolated from 56 infective or dead chickens of 20 egg laying farms in Kyung Buk province during the period from August to December 1994. 1. Among 416, 000 chickens of 92 flocks in 20 egg laying farms, 17, 360 chickens of 31 flocks were died of Fowl Typhoid. 2. Salmonella gallinarum was isolated from 56 chickens in liver and spleen, and then blood of infective chickens was positive to Pullorum antigen. 3, In the survey of gross lesion of 56 chickens, 43 chickens(76.8%) were swelled at liver, 39(69.6%) were swelled at spleen, 12(21.4%) were changed with bronze, 3(5.4% ) were hemorrhagic in peritoneal cavity. 4. In transmission pattern, 4 farms were outbreaked the entrance of chicken house at first, but the others were outbreaked at various place. They were transmitted at right and left directions in flock. 5. 2 farms confirmed at the early stage of infection were eradicated by removing infective chickens and administrating antibiotics, but 18 farms at chronic stage were not. 6. The biochemical properties of 112 Salmonella gallinarum from chickens were generally identical to those of the referance, but H$_2$S was not productive, cellobiose was fermentive. 7 Minimum inhibitory concentration(MIC) of 20 isolates was performed by using 21 antibiotics, MICs of Amikacin(Ak), Gentamicin(Gm), Kanamycin(Km), and Tetracycline (Tc) were below 1.6 ug/ml, Ampicillin(Am), Furazolidone(Fu) and Neomycin(Nm) were below 3.1 ug/ml, Cephalothin(Ce), Cefazoline(Cf) and Chloramphenicol(Cm) were below 6.3 ug/ml, Nalidixic acid(Na), Polymyxin(Po) and Rifampicin(Rf) were below 12.5 ug/ml, Penicillin (Pm) was below 25 ug/ml, Colistin(Co) and Streptomycin(Sm) were below 50 ug/ml, Sulfamerazine(Sr) and Sulfamethazine (St) were below 200 ug/ml, Lincomycin(Lm) and Spiramycin(Sp) were below 400 ug/ml, Bacitracin(Ba) was below 800 ug/ml. 8. Among the 20 isolates, all(100%) of those were sensitive to Ak, Am, Ce, Cf, Cm, Fu, Gm, Km, Na, Nm, Po, Rf, Sr, St and Tc, but 6 isolates(30%) were resistent to Co, 20(100% ) to Ba, Lm, Pm, Sm, and Sp. The drug resistance patterns were simple which 6 strains were BaCoLmPmSmSp type, and 14 were BaLmPmSmSp type.

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Use of enzyme-linked immunosorbent assay (ELISA) for detection of toxoplasmosis in dogs (ELISA 법을 이용한 개 톡소플라즈마병의 조기진단에 관한 연구)

  • Suh, Myung-deuk;Joo, Hoo-don;Lee, Byung-hoon
    • Korean Journal of Veterinary Research
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    • v.31 no.4
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    • pp.491-500
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    • 1991
  • This study was conducted to detect the serum antibodies in the experimentally toxoplasma infected dogs and street dogs by use the of an enzyme-linked immunosorbent assay (ELISA). And this test was performed on the polystylene microplate by coating with the tachyzoites soluble antigen of T gondii (RH strain), incubated with sera diluted then, added with HPO-conjugated rabbit anti-dog IgG and o-phenylenediamine used as a substrate. Tachyzoites of T gondii harvested from mouse peritoneal cavity were purified by 30, 40 and 50% Percoll density gradient centrifugation and used as the source of antigen. The results obtained were summarized as follows; 1. The highest ratio of positive to negative (P/N ratio) was obtained at the level of $l{\mu}g/ml$ protein concentration of antigen with the 1/4000 dilution of the conjugate measured by checker-board titration. It was regarded as the optimum concentration of the antigen and conjugate. 2. Cut-off value in this IgG ELISA was 0.375 that was determined by mean absorbance (at 492nm) of IFA negative serum added with the dauble value of the standard deviation $(mean{\pm}2S.D.)$. 3. Serum ELISA IgG antibodies to T gondii in the exyerimentally infected dogs were detected firstly at the Week 3 after inoculation and the highest titer was recognized at the Week 4, 5 and 6 after inoculation. 4. Stability of the antigen absorbed in the microplates that were preserved at $4^{\circ}C$ and $-25^{\circ}C$ separately were prolonged up to 3 weeks and 10 weeks at $4^{\circ}C$ and $-25^{\circ}C$, respectively. However the reproducibility was not reliable after the preservation of 4 weeks and longer. 5. Positive rate of the specific antibodies in 312 test sera was 28.5% and there was no significant differences between the male (27.8%) and female (29.5%), respectively. 6. The IgG ELISA was proved to be a specific procedure for the detection of antibodies to canine toxoplasma infection and also evaluated as a screening test for the large scale of test samples in laboratory.

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Scanning electron microscopy on proliferative forms of Toxoplasma gondii and Sarcocystis species (Toxoplasma gondii와 Sarcocystis 원충(原蟲)의 증식형(增殖型)에 대(對)한 주사전자현미경적(走査電子顯微鏡的) 관찰(觀察))

  • Kang, Yung-bai
    • Korean Journal of Veterinary Research
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    • v.28 no.2
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    • pp.379-385
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    • 1988
  • For the comparison of surface fine structures in the proliferative forms of two major protoroan parasites, Toxoplasma gondii and Sarcocystis species in mammalian hosts, isolated from the artificially infected mice and from the naturally infected cattle, respectively, an SEM(Hitachi S-570) was applied to the fixed, dried and coated with gold ion on the microslide glasses. The tachyzoites of T gondii from the peritoneal cavity of the mouse showed the crescent-like feature and measured as $5.57{\mu}m$ in length and $2.33{\mu}m$ in width, while the bradyzoites of Sarcocystis species from the heart muscle of slaughtered cattle was banana-shaped and measured as $14.18{\mu}m$ in length and $2.85{\mu}m$ in width. On the surface of Sarcocystis species bradyzoite, a distinct elliptical micropore was identified in the high magnification observation of 60,000X, and it measured as $0.35{\mu}m$ in length and $0.18{\mu}m$ in width.

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Experimental studies on antitumor effects and immune responses of Banhabaekchulcheonmatang and Banhabaekchulcheonmatanggamibang (반하백출천마탕(半夏白朮天麻湯)과 반하백출천마탕가미방(半夏白朮天麻湯加味方)의 항암효과(抗癌效果)와 면역반응(免疫反應)에 관(關)한 실험적(實驗的) 연구(硏究))

  • Baek, Tae-Hyoun;Ryu, Bong-Ha;Park, Dong-Won;Ryu, Ki-Won
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.1 no.1
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    • pp.141-165
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    • 1995
  • In order to investigate the effects of Banhabaekchulcheonmatang and Banhabaekchulcheonmatanggamibang on antitumor effects after Sarcoma-180 cells transplantation into the peritoneal cavity or left groin in mice, and immune responses in mice induced by methotrexate, the extracts of its herbal medicines were orally administered for 14 or 21 days. Experimental studies were performed for measurance of mean survival days, tumor and body weight for antitumor effects, and delayed type hypersensitivity, hemagglutinin titer, hemolysin titer, rosette forming cells, natural killer cell activity and phagocytic activity for immune responses in ICR mice. Following results were obtained : 1. Mean survival time in Banhabaekchulcheonmatang and Banhabaekchulcheonmatanggamibang-treated group was significantly prolonged, as compared with the control group(P<0.010, P<0.005). 2. Tumor weight in Banhabaekchulcheonmatang and Banhabaekchulcheonmatanggamibang-treated group was significantly depressed, as compared with the control group(P<0.050, P<0.050). 3. Body weight in Banhabaekchulcheonmatang-treated group was significantly increased (P<0.050), but in Banhabackchulcheonmatanggamibang-treated group was slightly increased with no effectivenes, as compared with the control group. 4. Delayed type hypersensitivity in Banhabaekchulcheonmatang and Banhabaekchulcheonmatanggamibang-treated group was significantly increased, as compared with the control group(P<0.010, P<0.050). 5. Hemagglutinin titer in Banhabaekchulcheonmatang and Banhabaekchulcheonmatanggamibang-treated group was significantly increased, as compared with the control group(P<0.050, P<0.050). 6. Hemolysin titer in Banhabaekchulcheonmatanggamibang-treated group was significantly increased (P<0.050), but in Banhabaekchulcheonmatanggamibang-treated group was slightly increased with no effectiveness, as compared with the control group. 7. Rosette forming cells in Banhabaekchulcheonmatang and Banhabaekchulcheonmatanggamibang-treated group was slightly increased with no effectiveness, as compared with the control group. 8. Natural Killer cell activity in Banhabaekchulcheonmatang and Banhabaekchulcheonmatanggamibang-treated group was slightly increased with no effectiveness, as compared with the control group. 9. Phagocytic activity in Banhabaekchulcheonmatanggamibang-treated group group was significantly increased (P<0.050), but in Banhabaekchulcheonmatanggamibang-treated group was increased with no effectiveness, as compared with the control group. According to the above results, it could be suggested that Banhabaekchulcheonmatang and Banhabaekchulcheonmatanggamibang have prominent antitumor effects, and enhance both cellular and humoral immunity.

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Early Gonadogenesis and Sex Differentiation in the Korean Rose Bitterling, Rhodeus uyekii

  • Goo, In Bon;Kim, Jung Eun;Kim, Myung Hun;Choi, Hye-Sung;Kong, Hee Jeong;Lee, Jeong Ho;Park, In-Seok
    • Development and Reproduction
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    • v.20 no.4
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    • pp.289-296
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    • 2016
  • This report describes the sex differentiation of the Korean rose bitterling, Rhodeus uyekii, from hatching to 170 days post-hatch (DPH) in relation to total length (TL), body weight (BW), and integral water temperature (IWT). The growth curve of TL from just hatching to 83 DPH was $5.144e^{0.045t}$ ($R^2=0.961$; t, time), and that of BW was $2.398e^{0.086t}$ ($R^2=0.725$). Primordial germ cells (PGCs) were observed at 17 DPH (7.9 mm TL, 3.74 mg BW, $374^{\circ}C$ IWT), and thereafter began to protrude into the peritoneal cavity. At 21 DPH ($9.2{\pm}0.14mm$ TL, $4.8{\pm}0.07mg$ BW, $462^{\circ}C$ IWT), some PGCs contained condensed chromatin and oocyte were observed in meiotic prophase. In contrast to the ovaries, which grew gradually after sexual differentiation, testes began multiplying at 25 DPH (10.1 mm TL, 5.42 mg BW, $550^{\circ}C$ IWT), when testicular differentiation was first identified, and multiplied continuously thereafter. At 33 DPH (11.2 mm TL, 10.5 mg BW, $726^{\circ}C$ IWT), the developing testes contained spermatogonia that exhibited mitotic activity. No spermatocyte or sperm cell was observed until 83 DPH (18.9 TL, 48.2 mg BW, $1,826^{\circ}C$ IWT). At 170 DPH (32.5 mm TL, 270.1 mg BW, $3,740^{\circ}C$ IWT), which was the end point of this study, the mature ovaries showed germinal vesicle breakdown, while the mature testes contained observable spermatocytes and sperm cells. These results allow us to identify the sex differentiation type of the Korean rose bitterling as differentiated gonochoristic.