• Natural Product Sciences
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• v.4 no.2
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• pp.62-67
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• 1998

As a part of search for new biologically active constituents from aloe, we have isolated a glycopeptide, called G1G1M1DI2, from the gel(G1) of Aloe vera. Chemical and spectroscopic evidence indicated that G1G1M1DI2 is a glycopeptide. The molecular weight of G1G1M1DI2 was about 5,500 daltons, and the carbohydrate and protein contents were 20.9% and 32.6%, respectively. Periodate oxidation and enzymic degradation gave peptide moiety and carbohydrate moiety, respectively. Carbohydrate moiety is composed of fucose, galactose, glucose and mannose in a molar ratio of 0.5:2.4;48.8:48.3. Peptide moiety is composed of fifteen amino acids, and glutamic acid and glycine were the major componants. The glycopeptide, G1G1M1DI2, stimulated thymidine uptake of SCC 13 cells about 6.5 times the control. This result suggests that this glycopeptide has a skin cell proliferating activity.

• Journal of Chest Surgery
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• v.28 no.8
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• pp.731-741
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• 1995

Cardiac valve allografts have been used as replacements for diseased valves and right ventricular outflow tract reconstruction, the long term follow-up of which has been reported satisfactory. For a good long-term result, it is essential that the allograft be viable at implantation. In this study, we aimed at preparing the cardiac valve allografts aseptically, preserving them at cold- and cryo-conditions, and testing the viability of the allografts after preservation by four methods. We tested the viability of the cardiac valve allografts preserved in cold refrigerated state[4$^{\circ}$C in nutrient media & in liquid nitrogen tank[cryopreservation under -149$^{\circ}$C for pre-planned time periods. The testing methods were 1 glucose utility test 2 tissue culture 3 thymidine uptake test and 4 histologic evidence by light microscopy. We observed no differences in the viability between cold- & cryo-groups and similar results among the methods for testing the viability. In conclusion, there was no difference in the viability between cold- and cryopreserved-allografts at least for 14 days of preservation. And glucose utility test and thymidine uptake test were satisfactory in the evaluation of the allograft viability, since they were easy and rapid with relatively quantitative results.

• YAKHAK HOEJI
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• v.43 no.3
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• pp.369-377
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• 1999

A new series of highly water soluble platinum(II) complexes {Pt(II)[1,3-bis(phenylthio) propane](trans- -1,2-diaminocyclohexane) (PC-1) and Pt(II)[1,3-bis-(phenythio)propane] cis-1,2-diaminocyclohexane(PC-2)} were synthesized, and characterized by their elemental analysis and by various spectroscopic techniques[infrared(IR), 13C-nuclear magnetic resonance (NMR)]. In vitro antitumor activity of new Pt(II) complexes was tested against P-388 and L-1210 mouse lymphocytic leukemia cell lines, PC-14 / P, PC-14/ADM and PC-14 / CDDP human pulmonary adenocarcinima, DU-145 human prostate carcinoma, HT-1376 human bladder carcinoma, ZR-75-1 human breast carcinoma, MKN-45/P and MKN-45/CDDP human gastric adenocarcinoma cell lines using colorimetric MTT[3-(4,5-dimethyl thiazol-2-yl)-2.5-diphenyltetrazoliumbromide] assay for cell survival and proliferation. PC-1 showed active against L-1210, P-388 leukemia, human lung, stomach, prostate, bladder and breast cancer cell lines, and the antitumor activity of these compounds were comparable or superior to those of PC-2 and displatin. The nephrotoxicities of PC-1 and PC-2 were found quite less than that of cisplatin using MTT and [3H] thymidine uptake in rabbit proximal tubule cells and human kidney cortical cells. Based on these results, this novel platinum (II) complex compound (PC-1) represents a valuable lead in the development of a new anticancer chemotherapeutic agent capable of improving antitumor activity and low nephrotoxicity.

• YAKHAK HOEJI
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• v.35 no.3
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• pp.154-164
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• 1991

These experiments were conducted to investigate the effects of Glycyrrhizae Radix extract(GR) on histamine synthesis, lymphocyte blastogenesis in C57BL/6J mice splenocytes, IL-1 production, $Ca^{2+}$ uptake by macrophage-like P388D$_{1}$ cells and plaque forming cell assay against SRBC. Histamine contents, lymphocyte blastogenesis, IL-1 activity, $Ca^{2+}$ uptake and plaque forming cell were determined by enzyme isotope method, [$^{3}$H]-thymidine incorporation, C3H/HeJ mouse thymocytes proliferation, the addition of 5 $\mu$Ci/ml $^{45}Ca^{2+}$ to P388D$_{1}$ cell suspension and assay to sheep red blood cell, respectively. Cytotoxicity, which was expressed as 50% mortality, was occurred by the addition of GR(10$^{-3}$g/ml). Histamine production in mouse spleen cell culture was significantly increased by 48 hour incubation added 0.25$\mu\textrm{g}$/ml of Con A. Con A-dependent T-lymphocyte proliferation was also enhanced by the addition of 0.25 $\mu\textrm{g}$/ml of Con A. GR depressed histamine contents at 10$^{-9}$~10$^{-4}$g/ml. and Con A (0.25 $\mu\textrm{g}$/ml) dependent T-lymphocyte proliferation at 10$^{-5}$~10$^{-4}$g/ml. IL-1 activity was significantly decreased by 10$^{-8}$~10$^{-4}$g/ml of GR. $Ca^{2+}$ uptake was not changed by GR, but antibody production markedly increased at 10.0~50.0 mg/kg of GR. From the above results, it is suggested that GR have immuno-regulatory action; GR decreased cell-mediated immune response and increased antibody production by B lymphocyte at high doses.

• Toxicological Research
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• v.18 no.2
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• pp.175-181
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• 2002

Allergic contact dermatitis (skin sensitization) may be caused by a wide variety of chemicals. A murine local lymph node assay (LLNA) has been developed as an alternative to guinea pig models for assessing the contact sensitization potential of chemical. This study was carried out to evaluate the skin sensitization potential for chemicals in Balb/c mice by LLNA. Contact allergen, dinitrochlorobenzene (DNCB), respiratory allergen, toluene diisocyanate (TDI) and a weak allergen, $\alpha$-hexlycinnamaldehyde (HCA) were wed as positive chemicals and irritant, sodium lauryl sulfate(SLS) also wed as a reference chemical in this study. The weights of lymph node in the mice treated with DNCB, TDI, and HCA were increased compared to vehicle control. There was a significant increase in lymph node weight of mice treated with high concentration of SLS compared to vehicle control. The stimulation index (SI) of Lymph node cell in the mice treated with DNCB, TDI, and HCA revealed over three-fold increase compared to vehicle control by $3^H$-thymidine uptake. All allergens correctly identified in this LLNA study wing Balb/c mice. These results suggest that LLNA wing Balb/c mice could be a useful method for screening the allergenic potential of chemicals. The expression of IL-2 mRNA was slightly increased in draining auricular lymph node cell of the mice treated with TDI and HCA by RT-PCR. However the IL-2 levels in DNCB and SLS of treated animals were not significantly changed.

• The Journal of Internal Korean Medicine
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• v.12 no.2
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• pp.96-112
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• 1991

Attempts were made to see the antitumor effects of Sigbunhwan widely used in Oh-jug(五積) employing tumor cells Lines such as K562 derived from erythroleukemia, Raji from lympoma and MO-4 from blastogenic tumor. Different concentrations of Sigbunhwan and combined therapy of Sigbunhwan and Bigihwan were treated to those tumor cells lines and then live cells were counted by Trypan blue assay and $^{3}H-Thymidine$ uptake assay. The results obtained were as follows. 1. $^{3}H-Thymidine$ uptake of various tumor cells lines when treated with high concentrations of Sigbunhwan for 48hours showed that the rate of DNA synthesis decreased 76% to 90% by the treatment of 1% Sigbunhwan but this inhibition was rather decreased when Sigbunhwan concentration was increased to 10, 15 and 20%.(Fig 3) 2. When Sigbunhwan was combined with Bigihwan which was also an antitumor drug, the effectiveness of tumor cells dealth was somewhat inceased showing a generally similar pattern to that of Bigihwan alone used.(Fig 4) This combination therapy also showed that higher concentrations of antitumor agent were no more·effective or rather harmful according to the tumor cells lines having different growth rate.(Fig 5,6) 3. The antitumor effects of combined Sigbunhwan and Bigihwan was decreased if the concentrations of this combination therapy was increased to 10 times showing relatively sluggish decrease in K562 and MO-4 but a sharp inhibitory effect in Raji which grows slowly.(Fig 7). 4. When Sigbunhwan was treated at low concentrations, K562 was more inhibited by 0.75% to 1.0% of Sigbunhwan while Raji was more inhibited by 0.25% to 0.5% of that.(Fig 8) 5. When Sigbunhwan was treated together with Bigihwan at low concentrations, the tumor cells death rate was 75% to 89% in Baji, 31% to 95% in MO-4 and 41 to 89% in K562, showing this combination therapy was more effect to Raji derived from lymphoma.(Fig 9) 6. The number of live tumor cells was correlated with optical density of MTT assay when measured with 2% Sigbunhwan treatment to tumor cells lines for 24 hours.(Fig 10) 7. 7 days treatment of 0.25% Sigbunhwan was compared with one day treatment of 1% suggesting long term treatment more effective.(Fig 11)

• The Journal of Internal Korean Medicine
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• v.24 no.1
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• pp.134-143
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• 2003

Objectives : We performed this study to understand the molecular basis of the antitumor effect of Saussurea lappa, Pharbitis nil, Plantago asiatica and Taraxacum mongolicum, which have been used for cancer treatment in Korean traditional medicine. Design: We analyzed, the effect of these medicinal herbs on proliferation and apoptosis of tumor cells and its association with gene expression, We performed semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR) analysis of cell cycle- and apoptosis-related genes using a gastric cancer cell line AGS. Results : Cell counting assay and $[^3H]thymidine$ uptake analysis showed that Saussurea lappa and Pharbitis nil strongly inhibit cell proliferation of AGS in a dose-dependent manner. Interestingly, gene espression assay revealed that mRNA espression levels of c-Jun, c-Fos, c-Myc, and Cyclin D1 were markedly decreased by Saussurea lappa and Pharbitis nil. Furthermore, Saussurea lappa was identified to activate expression of the p53 tumor suppressor and its downstream effector $p21^{Wafl}$, which leads to $G_1$ cell cycle arrest and apoptosis. These observations suggest that the anticancer effect of Saussurea lappa and Pharbitis nil might be associated with their regulatory capability of tumor-related gene expression.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.27 no.4
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• pp.314-320
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• 2001

Matrix metalloproteinases have long been viewed as ideal candidates for proteinases that enables tumor cells to permeated basement membrane defenses and invade surrounding tissue. There is growing evidence that the MMPs have an expanded role, as they are important for the creation and maintenance of a microenvironment that facilitates growth and angiogenesis of tumors at primary and metastatic sites. MT-MMPs are not secreted but instead remaining attached to cell surfaces. Although not all of the MT-MMPs are fully characterized, MT-MMPs have important role in localizing and activating secreted MMPs. The MMP genes are transcriptionally responsive to a wide variety of oncogene, growth factors, cytokine, and hormones. Currently, a number of MMP inhibitors are being developed and some have reached clinical trials as anti-metastatic or anti-cancer therapies. MT1-MMP is involved in the activation of proMMP-2. MT1-MMP is significant not only as a tumor marker but as a new target for chemotherapy against cancer. The purpose of this study was to evaluate the effects of protein kinase C inhibitor(genistein) on the proliferation of HT1080 and expression of MT1-MMP mRNA. Human fibrosarcoma cell line HT1080 was cultured and divided 2 groups. The experimental group was treated with $100{\mu}M$ genistein and incubated 12h, 24h for $[3^H]-thymidine$ uptake assay and northern hybridization individually. And the control group was treated with same amount of PBS for the above procedures. $[3^H]-thymidine$ incorporation was measured with ${\beta}$ ray detector. And RT-PCR and northern blotting for MT1-MMP mRNA was performed. The results were as follows 1. $[3^H]-thymidine$ uptake was reduced in experimental group with statistical significance. 2. MT1-MMP mRNA expression was significantly reduced in experimental group. These results showed that protein kinase C inhibitor (genistein) inhibited proliferation of HT1080 and almost completely blocked transcription of MT1-MMP mRNA. So, it is possible to use the protein kinase inhibitor (genistein) as anti-metastatic and anti-proliferative agent.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.12 no.2
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• pp.20-52
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• 1999

In order to investigate the effect of Naetakchungumsankamibang(NTCGS) water extract on the skin tumor induced by 3-MCA and immunological responses in mice, the cytotoxicity against SK-MEL-2 cells and total number of tumors induced by 3-MCA were measured. The numbers of WBC, platelets and RBC, plaque forming cells, hemagglutinin titer, hemolysis titer, carbon clearance, proliferation of splenocyte by thymidine uptake assay, splenic leukocyte by FACS analysis and $TNF-{\alpha}$ were also measured for the evaluation of the immunological responses. The results were obtained as follows: 1. In cytotoxicity against SK-MEL-2 cells, concentration inhibiting cell growth up to below $20\%$ of control was recognized at 1mg/ml of NTCGS. 2. In Inhibitory effect on the skin tumor induced by 3-MCA, the results showed a strong inhibitory effect of NTCGS. 3. In hematological changes in the tumor bearing mice, the numbers of WBC decreased significantly in NTCGS treated group as compared with control. 4. In hematological changes in the tumor bearing mice, the numbers of platelets increased significantly in NTCGS treated group as compared with control. 5. In hematological changes in the tumor bearing mice, the numbers of RBC increased with no significance in NTCGS treated group as compared with control. 6. Effects of the plaque forming cells in the tumor bearing mice, NTCGS treated group exhibited a significant effect compared with control. 7. In terms of the effects on hemagglutinin titer, NTCGS treated group showed higher level than control, without significance. 8. In terms of the effects on hemolysis titer, NTCGS treated group showed higher level than control, without significance. 9. In terms of the effects on phagocytic index K in Balb/C mice, NTCGS treated group showed significant difference from control. 10. In terms of the effects on proliferation of splenocyte by thymidine uptake assay, NTCGS showed significant effect at the concentration of 0.5mg/ml. 11. In terms of the effects on splenic leukocyte of Balb/C mice by FACS analysis, NTCGS treated group showed significantly higher level of helper T cell, B cell and macrophage than in control. 12. In terms of the effects on the secretion of $TNF-{\alpha}$, the treated group showed significant effect at the concentration of 1mg/ml of NTCGS. Based on the results summarized above, NTCGS is considered to have antitumor activity and immunological responses against skin tumor, and to be usable fur the treatment.

• YAKHAK HOEJI
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• v.35 no.3
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• pp.174-181
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• 1991

These experiments were conducted to investigate the effects of glycyrrhizin(GL) and glycyrrhetinic acid(GA) on histamine synthesis, lymphocyte blastogenesis in C57BL/6J mice splenocytes, IL-1 production, $Ca^{2+}$ uptake by macrophage-like P388D$_{1}$ cells and plaque forming cell assay against SRBC. Histamine contents, lymphocyte blastogenesis, IL-1 activity, $Ca^{2+}$ uptake and plaque forming cell were determined by enzyme isotope method, [sup 3/H]-thymidine incorporation, C3H/HeJ mouse thymocytes proliferation, the addition of 5 $\mu$Ci/ml $^{45}$Ca$^{2+}$ to P388D$_{1}$, cell suspension and assay to sheep red blood cell, respectively. Cytotoxicity, which was expressed as 50% mortality, was occurred by the addition of GL(10$^{-3}$M) and GA(10$^{-4}$M). Histamine production in mouse spleen cell culture was significantly increased by the addition of 0.25 $\mu\textrm{g}$/ml of Con A, after 48 hour incubation. Con A dependent T-lymphocyte proliferation was also enhanced by the addition of 0.25 .mu.g/ml of Con A. The effects of GL on histamine contents and T-lymphocyte proliferation were significantly decreased at high dose (10$^{-5}$M), while IL-1 activity was remarkably suppressed by 10$^{-8}$~10$^{-4}$M of GL. $Ca^{2+}$ uptake was not changed, but antibody production was increased by GL(10 mg/kg). GA inhibited histamine contents at 10$^{-9}$~10$^{-7}$ and depressed Con A (0.25 $\mu\textrm{g}$/ml) dependent T-lymphocyte proliferation at 10$^{-7}$~10$^{-5}$M of GA, but increased suboptimal dose (Con A 0.1 $\mu\textrm{g}$/ml) at 10$^{-9}$~10$^{-7}$M of GA. IL-1 activity was suppressed by 10$^{-8}$~10$^{-4}$M of GA and $Ca^{2+}$ uptake was enhanced by 10$^{-9}$~10$^{-6}$ of GA, but antibody production was not changed by GA. From the above results, it is suggested that GL and GA have immuno-regulatory action. GL decreased cell-mediated immune response, and increased humoral immune response at high dose. On the other hand, low dose of GA enhanced cell-mediated immune response, while high doses of GA decreased humoral immune reaction.