• Applied Biological Chemistry
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• v.34 no.3
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• pp.231-234
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• 1991

In the present investigation we have studied the effect of bear's gall on mammalian cells and demonstrated that COS-7 cells, which were derived Monkey kidney cells, had shown almost same extent of growth with 78 hrs in 10% FCS Dulbecco's Modified Eagle's medium with bear's gall and without bear's gall. But the hybridoma cells which were fused murine myeloma cells and the rat spleen cells for monoclonal antibody production died almost within 48 hrs. To investigate the effect of biosynthetic mechanism, cDNA were transfected to COS-7 cells, and it was shown that cDNA-transfected COS-7 cell had produced 30-40% less the amount of recombinant protein than the medium without bear's gall.

• Korean Journal of Animal Reproduction
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• v.20 no.1
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• pp.53-61
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• 1996

This study was carried out to determine effectively the sex of rabbit embryos using H-Y antiserum. H-Y antiserum was obtained from inbred SD strain female rat which was immunized by injection of testis cell of inbred SD strain male rate into its spleen. The titer of antiserum was identified by sperm cytotoxicity test and culture of rabbit embryos with antiserum. The developed or undeveloped embryos were separated by exposure the embryos to the antiserum with H-Y antibody. Developed embryo were transferred to the recipients and sex of offspring were examined. 1. In the sperm cytotoxicity test, the rate of dead sperm showed no difference between two antisera from spleen and testis cell as antigens. It is confirmed that H-Y antibody in antiserum was absorbed by H-Y antigen in male rat spleen cells. 2. When rabbit morulae were exposed to antiserum and complement, the rate of embryos developed or arrested was 51 and 49% respectively and the rate was closely same as natural sex ratio of 50:50%. 3. When rabbit morulae were cultured for 12h in the medium containing antiserum produced by antigen of testis cell, the rate of embryos developed or arrested was 48 and 52% respectively and the rate was closely same as natural sex ratio of 50:50%. 4. Eighty rabbit embryos which were not affected by H-Y antiserum were transferred to four recipients. Two recipients were pregnant and born 13 pups among which 2 (14%) were male and 11 (86%) were female. In conclusion, existence of H-Y antibody in the serum from female rat immunized by injecting testis cell from newborn male rat to the spleen of the female rat was confirmed. When rabbitmorulae were exposed to H-Y antiserum and complement, about a half of embryos were developed to blastocysts. When the rabbit embryos not affected by H-Y antiserum were transferred, the rate of female offspring was 86%. Therefore, it was identified that most of embryos which were not affected by H-Y antiserum were female.

• Korean Journal of Animal Reproduction
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• v.17 no.4
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• pp.305-310
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• 1994

These expriments were carried out to investigate existence of H-Y antibody in the rat serum immunized against H-Y antigen from rat spleen cells and effect of H-Y antiserum on development of mouse male embryos. The results obtained were summerized as follows : 1. When mouse embryos were cultured for 48∼72 hrs in the Ham's F10 containing 16% of FBS(fetal bovine serum) or RNS(rat normal serum), percentages of embryos developed from 2, 4, 8 and 16-cell embryo to morulae were 20, 27, 94 and 100%, respectively, in FBS and 8, 7, 94 and 100%, respectively, in RNS. Eight to 16-cell embryos showed no difference in development rate between FBS adn RNS. 2. When 8∼16-cell mouse embryos were cultured for 24∼48 hrs in the Ham's F10 containing FBS, RNS＋GPC(guinea pig complement) and RAS(rat antiserum)＋GPC, proportions of embryos developed to the expanded blastocyst stage were 100, 82.4 and 52.1∼53.6%(ave.52.9), respectively, so that it was suggested that rat antiserum suppressed development of male embryos. 3. When 8∼16-cell mouse embryos were cultured for 24∼48 hrs in the Ham's F10 containing FBS, RNS, RNS＋GPC and RAS＋GPC, proportions of embryos developed to the expanded blastocyst stage were 94.5, 90.9, 82.3 and 47%, respectively, and the embryos developed in the medium containing RAS＋GPC seemed to be female. These results indicated that the antisera prepared through immunized against H-Y antigen from rat spleen cell, possessed H-Y antibody which supressed development of male embryos.

• Korean Journal of Pharmacognosy
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• v.43 no.2
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• pp.157-162
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• 2012

In order to investigate the immunostimulating effect of mycelia extract of Phellinus linteus (PLM) on human monocyte THP-1 and rat peritoneal macrophage cell, we examined measuring cytokine secretion (IL-6 and TNF-${\alpha}$). The production of IL-6 and TNF-a in human monocyte THP-1 was slight increased dose-dependently when the cells were challenged with PLM for 72 hrs. It was also observed that the treatment of PLM with LPS augmented the production of IL-6 and TNF-a in human monocyte THP-1. It was also observed that the treatment of PLM with LPS augmented the production of IL-6 and TNF-${\alpha}$ in human monocyte THP-1. The production of IL-6 and TNF-${\alpha}$ in rat peritoneal macrophage was significantly enhanced when the cells were treated PLM with LPS for 72 hrs. Moreover, the proliferation rate of rat spleen cells was increased in a dose dependent manner as the cells were treated with PLM and Concanavalin A.

• Journal of Pharmacopuncture
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• v.3 no.2
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• pp.79-97
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• 2000

Objectives : To investigate the effects of herb-acupunctures of Qi tonification herbs or formula on the rat immune depression induced by an anticancer drug, methotrexate (MTX) Methods: Animals were divided into 5 groups; Normal control group was not given any treatment. Immune depression was induced by oral administration of 1mg/kg MTX b.i.d for 4 days in Control, Sample I, II, and III groups. We treated on CV 4 (Guanyuan) with saline, Bojoongiggi-tang (Buzhongyiqi-tang), Ginseng Radix and Astragali Radix herb-acupunctures in Control, Sample I, II, and Ill groups, respectively. In both blood and spleen, the assessment of CD4+ T-cell count, CD8+ T-cell count, CD4/CD8 T-cell ratio was performed. Results: Here we show that 3 herb- acupuncture groups have an influence, to some extent, on CD4+ and CD8+ T-cell counts and CD4/CD8 T-cell ratio in both blood and spleen. Astragali Radix herb-acupuncture, in particular, was found to have significantly increased CD4+ T-cell count in blood and CD4/CD8 T-cell ratio in blood and spleen. Conclusions: The results of this study suggest that herb-acupunctures of Bojoongiggi-tang (Buzhongyiqi-tang), Ginseng Radix and Astragali Radix may have an influence over rat immune depression induced by MTX.

• Korean Journal of Animal Reproduction
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• v.10 no.2
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• pp.168-174
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• 1986

These experiments were carried out to clarify the optimalconditions and interspecific cross reaction of H-Y antibody activity. H-Y antiserum was prepared in inbred SD female rats and Balb/c female mice by repeated immunization of rat newborn testis homogemate, rat and mouse spleen cells obtained from males of same strain. The activity of H-Y antibody in antiserum was tested by ELISA and biological tests. The cross reactivity of H-Y antibody was confirmed by culturing mouse and rabbit embryos in medium containing H-Y antibody and complement obtained from rat and guinea pig, respectively. The optimal condition for the activity of H-Y antibody was also investigated by culturing embryos in medium with different pH and complement concentration. The results obtained in these experiments were summarized as follows: 1. The formation rates of H-Y antibody in rats immunized with newborn testis and spleen cell were 40.0 and 50.0% respectively, and that in mouse immunized with spleen cell was 48.4%. 2. The activity of H-Y antibody was not affected by pH in range of 6.5 to 8.0, and the same was true for the relative concentration of complement to the H-Y antibody. 3. Minimum time needed for the activity of H-Y antibody was confirmed to be 0.5 to 1 hour and 24 to 48 hours respectively for the zona free embryos and intact embryos. 4. When mouse and rabbit embryos were treated with H-Y antibody obtained from rat, 46.4 and 54.8% of embryos were retarded or destroyed. From these results it could be said that H-Y antibody had strong interspecific cross reactivity.

• Korean Journal of Pharmacognosy
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• v.18 no.2
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• pp.118-126
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• 1987

Natural Killer cells are considerd to play an important role in antitumor immune surveilance mechanism. In this study, 21 putative anticancer drugs selected from reference were assessed by evaluating the effect on rat Natural Killer cell activity (NKCA). All 21 herb drugs were extracted in boiling water, lyophilized, autoclaved, and then used for experiment. Culture supernatant of concanavalin-A (Con-A)-stimulated rat spleen cells as a source of lymphokine was also used as a control of comparison. Rat spleen cells were used as effector and NKCA was measured in 4hr $^{51}Cr-release$ assay against Yac-1 mouse lymphoma cell line. In order to determine the optimal conditions for NKCA augmentation, effector cells were treated with 3 different concentrations of each drug for 24, or 48 hrs before testing of NKCA, In optimal conditions determined from previous results, the effect of herb drugs on NKCA were assessed in 3 to 5experiments. NKCA was significantly enhanced by treatment with 4 herb drugs(Ponciri Fructus, Houttuyniae Herba, Aurantii Pericarpium, Nepetae Herba). Culture supernatant of Con-A-stimulated spleen cells also augmented the rat NKCA more significantly. The results show that 4 of the herb medicines supposed to display anticancer effect may have activity as a biological response modifier through augmentation of NKCA.

• BMB Reports
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• v.34 no.2
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• pp.156-165
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• 2001

Leptin is a circulating non-glycosylated protein that is mainly produced in adipocytes. Leptin acts in the brain to regulate food intake and energy expenditure. Previously we reported our success in the isolation of a partial cDNA of the long form of the leptin receptor, OB-Rb, from rat spleen, and showed that leptin might also play a role in peripheral immune organs. In the present study, for the first time, the complete coding region of OB-Rb cDNA was cloned from rat splenocytes, and its nucleotide sequence was determined. The cDNA was then further expressed in E. coli and mammalian cells, thereby confirming the functional integrity of this receptor. Prokaryotically overexpressed OB-R protein was then used as an immunizing antigen in BALE/c mice to produce leptin receptor-specific antibodies. By using them, we confirmed the cell surface expression of OB-Rb in transfected CHO cells. It is our belief that the reagents, as produced in this study, will be of great use in further studies of the biological role of rat leptin.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.6
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• pp.1576-1583
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• 2006

In order to investigate the effect of Goamshimshinhwan(GASSW) on SD rats with deteriorated immunity caused by methotrexate. Methotrexate was fed to the SD rats once a day for 4 days. After the immune responses of the at a dosage 1,000, 500 and 250mg/kg/10ml. and the changes on body weight and gains, spleen weight, total blood leukocyte numbers, total lymphocyte numbers, the percentage of B-cell, T-cell, CD3+CD+4 T-cell, CD3+CD8+ T-cell and CD4+/CD8+ T-cell ratios in the bolld and spleen were observed. In addition, the serum IL-2 levels and productivity of IL-2 of splenic cells were also demonstrated in this study. The changes on body weight were increased significantly in 100 and 500mg/kg of GASSW groups and the changes on body gain were increased significantly in 1000mg/kg of GASSW groups as compared with control group. The changes on the spleen weight (absolutely or relatively) were increased significantly in all GASSW groups as compared with control group. The total blood leukocyte numbers were increased significantly in 1000 and 500mg/kg of GASSW groups as compared with control group. The total lymphocyte numbers were increased significantly in all GASSW groups in the blood and increased significantly in 1000 and 500mg/kg of GASSW goups in spleen as compared with control group. The percentage of B-cell and T-cell were increased significantly in 1000mg/kg of GASSW groups in the blood and increased significantly in 1000 and 500mg/kg of GASSW groups in spleen as compared with control group. The percentage of CD3+CD4+ T-cell and the serum IL-2 levels and productivity of IL-2 of splenic cells were increased significantly in 100 and 500 mg/kg of GASSW groups in the blood and spleen as compared with control group. The percentage of CD3+CD8+ T-cell were increased significantly in 1000mg/kg of GASSW groups only in spleen as compared with control groups. The CD4+/CD8+ T-cell ratios were increased significantly in 1000 and 500mg/kg of GASSW groups only in the blood as compared with control group. Goamshimshinhwan(GASSW) has immuno-stimulating effect on SD rats with deteriorated immunity caused by methotrexate.

• The Journal of Internal Korean Medicine
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• v.25 no.4
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• pp.117-128
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• 2004

Objectives : To examine the effects of Gojineumja on white rats which deteriorated immunity caused by Methotrexate(MTX), first of all, MTX was fed to the rats once a day for 4 day. Methods : After the immune response of the rats are deteriorated, dried extracts of Gojineumja(GJE) mixed in water was fed to the white rats once a day for l4days. The next conclusion was made by examining the rates of B-cells and T-cells of the peripheral blood and the changes in rates of CD4+ T-cells and CD8+ T -cells of the blood sampled from the spleen and peripheral region. Especially the count of CD3+ CD4+ T-cells of the peripheral blood and the count of CD3+ CD4+ T-cells of the spleen the count of CD4+/ CD8+ T-cell of the peripheral blood and the spleen proved the significant effect of increasing immune responses statistically. Results :(1) The following are the summary of the results. (2) The percentage of B lymphocyte of peripheral blood was increased significantly in GJE group as compared with control group. (3) The percentage of CD3+ CD4+ T-cell of peripheral blood was increased significantly in GJE group as compared with control group. (4) The percentage of CD3+ CD8+ T-cell of peripheral blood was not different statistically. (5) The percentage of CD4+/ CD8+ T-cell of peripheral blood was increased significantly in GJE group as compared with control group. (6) The percentage of CD3+ CD4+ T-cell of spleen was increased significantly in GJE group as compared with control group. (7) The percentage of CD3+ CD8+ T-cell was not different statistically. (8) The percentage of CD4+ /CD8+ T-cell was not different statistically. Conclusions : Gojineumja has an effect of increasing immune responses on white rats with deteriorated immunity caused by MTX.