• The Journal of Internal Korean Medicine
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• v.24 no.2
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• pp.315-328
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• 2003

This study was performed to investigate the effect of Bunsimgieum on antitumor effect after sarcoma-180 cells transplantation into peritoneal cavity or left groin and immune responses on the depressed immunity induced by methotrexate in mice. The Bunsimgieum extract of 10mg/kg was orally administered 14 days for antitumor effects and 21 days for immune responses. 50% inhibitory concentration($IC_{50}$) of SUN-1, SUN-C4, and SUN-396 cancer cell, mean sunvival days and body weight of tumor bearing mice, and growth of tumor mass for antitumor effect; delayed type hypersentivity, hemagglutinin titer, hemolysis titer, rosette forming cells, natured killer cell activity, lymphocyte transformation, productivity of interleukin-2, and phagocytic activity for their immune responses were measured in ICR mice. Significance in antitumor effect is noted in the enlongation of mean life days and inhibition of tumor growth(p<0.01, respectively). Significance of immune responses is also noted in hemolysis titer, lymphocyte transfumotion, IL-2 productivity, phagocytic activity, and natural killer cell activity at E/T ratio 100:1(p<0.01, respectively). Significant in rosette cell formation was seen at dosage of 20mg/kg(p<0.01). However, Difference of body weight as antitumor effect, delayed type hypersensitivity, and hemagglutinin titer were not shown significantly. According to the above results, it could be suggested that Bunsimgieum has prominent antitumor and immunity enhancing effect.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.2 no.1
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• pp.57-73
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• 1996

These studies were consist of two sub-experiment. In order to study the effect of Banhahubaktang on the Cell-cytotoxicity In vitro. We had put through MTT Assay. In order to investigate the effects of Banhahubaktang on the ICR mice which had Abdominal tumor induced by Sarcoma-180 cell line, C57BL/6 mice which had pulmonary melanoma induced by B16 cell line. After Sarcoma-180 cell line and B16 cell line were transplanted, the extract of Banhahubaktang was orally administered to the mice to observe the extension of survival time of the mice, inhibition of solid tumor, inhibition of pulmonary melanoma metastasis. productivity of Interleukin-2, NK-Activity. The results were summarized as follows: 1. On the MTT assay, in case of $100{\mu}g/ml$ and $10{\mu}g/ml$ of Banhahubaktang concentration were inhibited cell viability significantly. But $1{\mu}g/ml$ of Banhahubaktang was tended to inhibit cell viability with no significance. 2. In the effect of life extension, Banhahubaktang treated group appeared to survive longer than the control group, but which were not significant. 3. In the effect of inhibit solid tumor, Banhahubaktang treated group appeared to decrease than the control group, but which were not significant. 4. In the effect of inhibit melanoma pulmonary metastasis. Banhahubaktang treated group appeared to inhibit than the control group significantly. 5. In the productivity of Interleukin-2, on 7 and 14 day, Banhahubaktang treated group increased than control group, which were significant. But on 21 day, test group and control group were much in common. 6. In the NK-Activity, Banhahubaktang treated group and control group were much in common.

• Journal of Food Hygiene and Safety
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• v.38 no.6
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• pp.430-441
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• 2023

Velvet antler is widely used as a traditional medicine, and numerous studies have demonstrated its tremendous nutritional and medicinal values including immunity-enhancing effects. This study aimed to investigate different deer velvet extracts (Sample 1: raw extract, Sample 2: dried extract, and Sample 3: freeze-dried extract) for proximate composition, uronic acid, sulfated glycosaminoglycan, sialic acid, collagen levels, and chemical components using ultra-performance liquid chromatography-quadrupole-time-of-light mass spectrometry. In addition, we evaluated the cytotoxic effect of the deer velvet extracts on BV2 microglia, HT22 hippocampal cells, HaCaT keratinocytes, and RAW264.7 macrophages using the cell viability MTT assay. Furthermore, we evaluated acute toxicity of the deer velvet extracts at different doses (0, 500, 1000, and 2000 mg/kg) administered orally to both male and female ICR mice for 14 d (five mice per group). After treatment, we evaluated general toxicity, survival rate, body weight changes, mortality, clinical signs, and necropsy findings in the experimental mice based on OECD guidelines. The results suggested that in vitro treatment with the evaluated extracts had no cytotoxic effect in HaCaT keratinocytes cells, whereas Sample-2 had a cytotoxic effect at 500 and 1000 ㎍/mL on HT22 hippocampal cells and RAW264.7 macrophages. Sample 3 was also cytotoxic at concentrations of 500 and 1000 ㎍/mL to RAW264.7 and BV2 microglial cells. However, the mice treated in vivo with the velvet extracts at doses of 500-2000 mg/kg BW showed no clinical signs, mortality, or necropsy findings, indicating that the LD50 is higher than this dosage. These findings indicate that there were no toxicological abnormalities connected with the deer velvet extract treatment in mice. However, further human and animal studies are needed before sufficient safety information is available to justify its use in humans.

• Journal of Acupuncture Research
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• v.19 no.4
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• pp.56-73
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• 2002

Objective : We need to develop a new treatment method which can curve cancer growth and enhance immunity of patients with various kinds of cancer more safely and effectively, for conventional anticancer treatment has lots of problems to be overcomed, in other words, Its efficacy can be recognizible but it doesn't actually give aid to patients due to its side effects. This study was taken up to evaluate the anticancer and immune-enhancing effect of Scutellariae Barbatae Herba, Alli bulbus, Oldenlandiae Herba(SAO) Herbal acupuncture. Methods : SAO Herbal acupuncture solution was made from Scutellariae Barbatae Herba, Alli bulbus, Oldenlandiae Herba by decoction. Experimental group was divided into normal(N), control(TC, cancer group induced by S 180), high and low concentration SAO complex Herbal acupuncture group. In the high and low concentration SAO complex Herbal acupuncture group, SAO Herbal acupuncture solution was injected, on the left and right Chok-samni(足三里, ST36) of ICR-male S 180 rats alternatively, by 200mg/kg and 100mg/kg respectively. In vitro, S 180 was cultured with $200{\mu}g$ and $500{\mu}g$ of SAO Herbal acupuncture solution. In each experimental group, we examined the effect of SAO complex Herbal acupuncture on body weight, antitumor, organ weight, activity of macrophage, activity of B cell, spleen cell division, IL-2 production and population of lymphocytes. Results : 1. In Body weight, no significant change was shown, but In solid cancer weight, the high concentration SAO complex Herbal acupuncture group showed signigicant(P<0.05) decrease and significant(P<0.05) increase in the weight of kidney, compared with control group. 2. In activity of macrophage, low concentration SAO complex Herbal acupuncture group showed significant(P<0.01) increase, but in vitro, there was no significant increase, compared with control group. 3. In activity of B cell, high and low concentration SAO complex Herbal acupuncture group showed no significant decrease, but in vitro, low concentration SAO complex Herbal acupuncture group showed significant(P<0.01) increase, compared with control group. 4. In spleen cell division, high and low concentration SAO complex Herbal acupuncture group had no significant influence on spleen cell division induced by Co A, meanwhile, it was found that macrophge promote spleen cell division in low concentration SAO complex Herbal acupuncture group(P<0.05), compared with control group. 5. In IL-2 production, high concentration SAO complex Herbal acupuncture group showed significant((P<0.05) increase, compared with control group. 6. In population of lymphocytes, high concentration SAO complex Herbal acupuncture group showed significant increase of CD3+(P<0.05), CD4+(P<0.05), CD3+ and CD4+ T cell(P<0.01) and B cell(P<0.05), while low concentration SAO complex Herbal acupuncture group showed significant increase of CD4+(P<0.05), CD8+ T cell(P<0.05) and B cell(P<0.01), compared with control group. Conclusion : SAO Herbal acupuncture inhibited cancer growth and enhanced immunity.

• Clinical and Experimental Reproductive Medicine
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• v.25 no.1
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• pp.77-86
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• 1998

The uptake of glucose for metabolism and growth is essential to most animal cells and is mediated by glucose-transporter (GLUT) proteins. The aim of this study was to determine which class of glucose transporter molecules was responsible for uptake of glucose in the mouse early embryo and at which stage the corresponding genes were expressed. In addition, co-culture system with vero cell was used to investigate the effect of the system on GLUT expression. Two-cell stage embryos were collected from the superovulated ICR female and divided into 3 groups. As a control, embryos were cultured in 0.4% BSA-T6 medium which includes glucose. For the experimental groups, embryos were cultured in either co-culture system with vero cells or glucose-free T6 medium supplemented with 0.4% BSA and pyruvate as an energy substrate. 2-cell to blastocyst stage embryos in those groups were respectively collected into microtubes (50 embryos/tube). Total RNA was extracted and RT-PCR was performed. The products were analysed after staining ethidium bromide by 2% agarose gel electrophoresis. Blastocysts were collected from each group at l20hr after hCG injection. They were fixed in 2.5% glutaraldehyde, stained with hoechst, and mounted for observation. In control, GLUT1 was expressed from 4-cell to blastocyst. GLUT2 and GLUT3 were expressed in morula and blastocyst. GLUT4 was expressed in all stages. When embryos were cultured in glucose-free medium, no significant difference was shown in the expression of GLUT1, 2 and 3, compared to control. However GLUT4 was not expressed until morular stage. When embryos were co-cultured with vero cell, there was no significant difference in the expression of GLUT1, 2, 3 and 4 compared to control. To determine cell growth of embryos, the average cell number of blastocyst was counted. The cell number of co-culture ($93.8{\pm}3.1$, n=35) is significantly higher than that of control and glucose-free group ($76.6{\pm}3.8$, n=35 and $68.2{\pm}4.3$, n=30). This study shows that the GLUT genes are expressed differently according to embryo stage. GLUTs were detectable throughout mouse preimplantation development in control and co-culture groups. However, GLUT4 was not detected from 2- to 8-cell stage but detected from morula stage in glucose-free medium, suggested that GLUT genes are expressed autocrinally in the embryo regardless of the presence of glucose as an energy substrate. In addition, co-culture system can increase the cell count of blastocyst but not improve the expression of GLUT. In conclusion, expression of GLUT is dependent on embryo stage in preimplantation embryo development.

• Clinical and Experimental Reproductive Medicine
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• v.24 no.2
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• pp.233-239
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• 1997

Embryos of most mammalian species grown in vitro would undergo developmental arrest at the approximate time of genomic activation. Stage-specific cell block and the resulting rapid loss of embryo viability in conventional culture media have limited the duration for which embryos may be cultured prior to transfer. As a result, embryos are usually transferred to the uterus at the 4-to 8-cell stage to avoid the loss of viability associated with long-term in vitro culture. Early transfer has led to asynchrony of the endometrium-trophectoderm interaction at the time of implantation and a resultant reduction in the rate of implantation. To overcome these problems, a variety of co-culture systems has been devised in which embryos can develop for a longer period prior to embryo transfer. Vero cells, derived from African green monkey kidney, share a common embryologic origin with cells from the genital tract. In addition, they are potentially safe to use, since they are highly controlled for viruses and other contaminants. Therefore, co-culture using Vero cells has been widely utilized to enhance embryo viability and development, although not without controversies. We thus designed a series of experiments to demonstrate whether Vero cells do indeed enhance mouse embryo development as well as to compare the efficacy of co-culturing mouse 1-cell embryos on Vero cell monolayer in both Ham's F-10 and human tubal fluid (HTF) culture media. 1-cell stage ICR mouse embryos were cultured either in the presence of Vero cells (Group A) or in conventional culture medium alone (Group B). In Ham's F-10 significantly more 3-to-8cell embryos developed in group A than group B (59.8 versus 10.0%; p<0.01). In contrast, there was no significant difference in embryonic development both group A and group B in HTF. However, significant differences were noted only in later embryonic stage (13 and 0%; p<0.05 of group A and B respectively, hatching or hatched). In Ham's F-10, we also could observe the beneficial effect of Vero cell on hatching process (70.7 and 42.1%; p<0.05 of group A and group B respectively).

• Tuberculosis and Respiratory Diseases
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• v.63 no.3
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• pp.251-260
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• 2007

Purpose: An imbalance of cell proliferation and cell apoptosis is an important mechanism in carcinogenesis. Capase 3, survivin and p53 have been identified as important members of the apoptotic related proteins. This study evaluated the proliferating cell nuclear antigen(PCNA), apoptosis, apoptotic related protein such as capase 3, survivin and p53 using urethane-induced mouse lung carcinogenesis, which provides reproducible steps from hyperplasia to adenocarcinoma. Methods: Urethane was administered to the ICR mice through an intra-peritoneal injection, The mice were sacrificed at 5, 15, and 25 weeks after urethane intervention. The sequential morphological changes and immunohistochemical expression of PCNA, apoptosis, capase 3, survivin, and p53 were examined during mouse lung carcinogenesis. Results: During carcinogenesis, the sequential histological changes were observed from hyperplasia of type II pneumocytes, to anadenoma, and ultimately to an overt adenocarcinoma. The PCNA Labeling index (LI) was 9.6% in hyperplasia, 23.2% in adenoma, and 55.7% in adenocarcinoma, respectively. The apoptotic LI was 0.24% in hyperplasia, 1.25% in adenoma, and 5.27% in adenocarcinoma. A good correlation was observed between the PCNA LI and apoptotic LI. The expression of caspase 3 was remarkable- i.e., 46.7% in adenocarcinoma, in contrast to 15% in hyperplasia and 16% in adenoma. Survivin was detected weakly in the alveolar hyperplasia and showed an increasing expressional pattern in adenoma and adenocarcinoma. p53 expression was detected only in the adenocarcinoma lesions with an expression rate of 13.3%. The level of caspase 3 expression correlated with the increase in the apoptotic index. The positive expression of caspase 3 was associated with an increased apoptotic index. Conclusions: These results suggest that the PCNA LI and apoptotic LI might be useful markers for evaluating the risk of a malignant transformation. In addition, caspase, survivin and p53 might play a role in the early and late steges of urethane-induced mouse lung carcinogenesis.

• Journal of Physiology & Pathology in Korean Medicine
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• v.27 no.1
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• pp.83-91
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• 2013

Pruritus is a unpleasant symptom in the skin that provokes the act of or desire to scratch. Mast cells are important effector cells in allergic reactions such as pruritus and inflammation. The purpose of this study was undertaken to investigate the synergic anti-pruritic and anti-inflammatory effects of Scutellariae Radix (SB) plus Flos Loncerae (FL) extracts in rat peritoneal mast cells (RPMCs), pruritogen-induced scratching mice and 2,4-dinitrofluorobenzene (DNFB)-induced allergic mice. We investigated the effect of SB, FL and SB plus FL extracts on the production of tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-$1{\beta}$, and histamine in RPMCs, on the scratching behavior in ICR mice, and skin clinical serverity and inflammatory mediators in DNFB-induced allergic hairless mice. RPMCs stimulated with PMA plus A23187 or compound 48/80 significantly increased TNF-${\alpha}$, IL-$1{\beta}$ or histamine production compared with media control. However, TNF-${\alpha}$ IL-$1{\beta}$ or histamine levels increased by PMA plus A23187 or compound 48/80 treatment were significantly inhibited by SB, FL in a dose-dependent manner. Especially, SB plus FL pretreatment had a synergic inhibitory effects on stimulator-induced cytokines (TNF-${\alpha}$ and IL-$1{\beta}$) and histamine production. Moreover, SB plus FL administration had a synergic inhibitory effects on the scratching behavior induced by pruritogen (compound 48/80, histamine, serotonin, substance P) in ICR mice. Furthermore, SB plus FL administration had a synergic inhibitory effects on skin damage, inflammatory mediators, leukocyte and mast cell infiltration induced by DNFB in hairless mice. These results suggest that SB plus FL administration has a potential use as a medicinal plant for treatment against itching and inflammation-related skin disease.

• Clinical and Experimental Reproductive Medicine
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• v.29 no.3
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• pp.201-207
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• 2002

Objective: To observe the capability of fertilization and embryo development including blastocyst formation of the oocytes in simple media after thawing of the cryopreserved cumulus-free mouse oocytes by vitrification method. Methods: Oocytes were collected from 5 to 6 weeks old ICR female mice, and were denuded from the cumulus cells by 0.1% hyaluronidase. Recovered mature oocytes in study group were cryopreserved by vitrification method using EM grid for $5{\sim}7$ days. In brief, oocytes were exposed in dPBS containing 1.5 M EG and 5.5 M EG+1 M sucrose for 2.5 minutes and 20 seconds each, and then executed vitrification by plunging in LN2 after loading on EM grid. Thawing treated by exposure of 1, 0.5, 0.25 and 0.125 M sucrose solution for 2.5 minutes each in order and used for experiments. Spermatozoa aspirated form the epididymis of 12 weeks old ICR male mice were used for insemination after capacitation. T6 media containing 0.4% BSA were used for fertilization and development. Results: Survival and fertilization rates after thawing were 76.9% and 79.6% respectively. Fertilization rate was lower (p<0.005) than that of control group (92.9%). There was no difference in embryo developmental rates from 2-cell to morula, however, the blastocyst formation rate and mean cell numbers of blastocysts in study group (63.3%, $58.9{\pm}9.2$) were lower compared with those of control group (76.1%, $63.5{\pm}8.9$). Conclusion: Vitrification is an effective method for mouse mature oocyte cryopreservation with high survival and fertilization rate after thawing. And in simple media, fertilization rates and embryo development of frozen-thawed mouse oocytes are satisfactory.

• Journal of Korean Society of Forest Science
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• v.99 no.1
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• pp.16-23
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• 2010

Experiments were performed for investigate the immune activities on human B and T cell growth and secretion of their cytokines. Also, antibodies in serum were investigated in female ICR mouse by feeding the extracts of Acer mono at dose of 30 and 100 mg/mL of one day orally for 15 days. The cytotoxicity of the samples on normal human cell (HEL299) was below 24.15% in adding the all extracts. Both human immune B and T cells were increased up to about 40% by the high pressure extraction process. The secretion of cytokines (IL-6, TNF-${\alpha}$) on human B and T cells were increased up to 20-50% by adding the high pressure extraction process compare to the hot water extraction process. Also, total serum IgG levels increased by feeding Acer mono extacts. It can be conclude that optimum condition for efficient extraction of Acer mono as functional materials is solvent extraction process using water with high pressure at below $100^{\circ}CA$ than typical process.