• Korean Journal of Pharmacognosy
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• v.23 no.3
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• pp.158-170
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• 1992

The studies were conducted to investigate the combined effects of several combined preparation of crude drugs and mitomycin C(MMC). The combined effects on the proliferation of Molt-4 cells and activation of human lymphocytes were estimated by MTT colorimetric assays. The drugs itself enhanced the proliferation of Molt-4, but the inhibitory action of MMC was not affected by the combined treatment of the drugs and MMC. Among 9 kinds of the drugs, Sip Jeon Dae Bo Tang(SDT), Saeng Maek San(SMS) and Kwi Bi Tang(KBT) did not inhibit the action of MMC, but activated lymphocytes. When the mice were treated by MMC, the number of leukocytes was decreased significantly at the 1st day, but recovered at the 7th day. In the groups of MMC treated with SDT or KBT, the number of leukocytes was increased significantly than the group of MMC treated only at the 3rd day. The combined treatment of SDT, SMS and MMC retained the body weight of mice at the level of normal mice. The SDT, SMS and KBT did not change the number of plaque forming cells(PFC) and the proliferation of T cells. The combined treatment of SDT and MMC increased the number of PFC significantly than the MMC treated group. The combined treatment of SDT, SMS, KBT and MMC increased the T cell proliferation significantly than the MMC treated group. In conclusion, it is suggested that SDT, SMS and KBT can recover the side effects of MMC, such as weight loss, leukopenia and immunosuppression, without any intercalating the anti-proliferative action of MMC in vivo.

• The Journal of Internal Korean Medicine
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• v.15 no.1
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• pp.60-79
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• 1994

The studies were conducted to investigate the combined effects of Tonics and Mitomycin C(MMC). The effects of Tonics and MMC on the proliferation of Molt-4 cells, human leukemic cell line, and activation of human lymphocytes were estimated by MTT colorimetric assays. Selected medicines among 9 kinds of Tonics by results of MTT assays were treated with MMC in mice. The Tonics itself enhanced the proliferation of Molt-4, but the anti-proliferative effect of MMC was not intercalated by the combined treatment of Tonic and MMC. Inhibitory action of MMC was augmented by Sa Kun Ja Tang(SKT). This result was due to the inhibition of DNA synthesis. Among 9 kinds of Tonics, Sip Jean Dae Bo Tang(SDT), Saeng Maek San(SMS) and Kwi Bi Tang(KBT) did not inhibit the action of MMC, but activated lymphocytes. When the mice were treated by MMC, the number of leukocytes was decreased significantly at the 1st day, but recovered at the 7th day. In the groups of MMC treated with SDT or KBT, the number of leukocytes was increased significantly than the group of MMC treated only at the 3rd day. Combined treatment of the Tonics(SDT, SMS) and MMC retained the body weight of mice at the level of normal mice. SDT, SMS and KBT did not change the number of plaque forming cells(PFC), but MMC treated group decreased the number of PFC. The combined treatment of MMC and SDT increased the number of PFC significantly than the MMC treated group. SDT, SMS and KBT did not influence the proliferation of T cells, but MMC treated group decreased the proliferation of T cells. The combined treatment of MMC and those tonics increased the T cell proliferation significantly than the MMC treated group. In conclusion, the results presented in this paper suggest that SDT, SMS and KBT can recover the side effects of MMC, such as weight loss, leukopenia and immunosuppresion, without any intercalating the anti-proliferative action of MMC in vivo.

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.2
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• pp.67-75
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• 2002

The human complement receptor type 1 (CR 1, C3 b/C4b receptor) is a polymorphic membrane glycoprotein expressed on human erythrocytes, peripheral leukocytes, plasma and renal glomerular podocytes, which consists of transmembrane and cytoplasmic domains with 30 repeating homologous protein domains known as short consensus repeats (SCR). CR1 has been used as an inhibitor for inflammatory and immune system for the past several years. Recently; it is reported that CRl was found to suppress the hyper-acute rejection in xeno-transplantation and can be used to cure autoimmune diseases. A soluble form of CRl, called sCRl, is a recombinant CRl by cleaving the transmembrane domain at C-terminus and has been expressed in Chinese Hamster Ovary (CHO) cells. Several purification methods for sCR1 from CHO cells have been reported, but most of them require complicated steps at high cost. Moreover, such methods are mostly performed under the pH condition apt to denaturing sCR1 and causes sCRl losing its activity. We here report a rapid and efficient method to purify sCR1 from CHO cell. The new method consists of a two-stage of cell culture by cultivating cells in serum medium followed by serum-free medium, and a two-stage of column purification by means of heparin and gel filtration column chromatography. By using this novel method, sCR1 can be purified in a simple and effective way with high yield and purity, furthermore, the purified sCR1 was confirmed to retain its activity to suppress the complement activation in vivo and ex vivo.

• Animal cells and systems
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• v.10 no.1
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• pp.15-20
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• 2006

Allergic inflammation is thought to be a Th2 cell-dominant immune response during which tissue-resident fibroblasts produce chemokines which contribute to the recruitment of migratory leukocytes to sites of tissue injury. Thymus and activation-regulated chemokine (TARC; CCL17) is a potent member of the CC chemokine family and a selective chemoattractant for Th2 cells. In order to study the regulatory profiles of TARC production by $TNF-{\alpha}$, $IFN-{\gamma}$, and Il-4 in human normal skin fibroblast, CCD-986sk cell line was used. The expression of TARC protein was measured using ELISA, and mRNA level was detected by RT-PCR. The combination of $TNF-{\alpha}$ and IL-4 induced a time-and dose-dependent synergistic increase in the expression of TARC at both protein and mRNA levels in the cultured human skin fibroblasts. Exposure of the cells to single cytokine had no effect on TARC expression. The high concentration (100 ng/ml) and long incubation time (72 h) of $IFN-{\gamma}$ further enhanced the TARC production induced by $TNF-{\alpha}$/lL-4 in the skin fibroblast. This synergistic effect of Th1 and Th2 type cytokines on TARC production by skin fibroblasts may contribute to the inflammatory cell infiltration and tissue damage with allergic inflammation.

• Restorative Dentistry and Endodontics
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• v.27 no.5
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• pp.463-472
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• 2002

Bacterial lipopolysaccharide (LPS) plays a major role in stimulating the synthesis and release of the principal osteoclast-activating cytokines, namely, interleukin 1 and tumor necrosis factor-$\alpha$ from immune cells. Although rnonocytes/macrophages are the main producers of these cytokines, recent evidence has indicated that polymorphonuclear leukocytes (PMN) have the ability to release IL-1 and TNF-$\alpha$. Calcium hydroxide has been shown to be an effective medicament in root canal infections, reducing the microbial titre within the canal. It has been proposed that the therapeutic effect of Ca(OH)$_2$ may also be the result of direct inactivation of LPS. The purpose of this study was to investigate whether treatment of Porphyromonas endodontalis LPS with calcium hydroxide alters its biological action as measured by human PMN secretion of IL-1 and TNF-$\alpha$, and it was compared with Escherichia coli LPS. P. endodontalis ATCC 35406 was cultured in anaerobic condition, and LPS was extracted using the hot-phenol water extraction method and purified. Purchased E. coli LPS was also purified. 100 $\mu\textrm{g}$/ml of each LPS in pyrogen free water were incubated with 25mg/ml Ca(OH)$_2$ at 37$^{\circ}C$ for 7 days. The supernatants were subjected to ultrafiltration, and the isolates were lyophilized and weighed. PMNs were obtained from peripheral blood by centrifugation layered over Lymphoprep. The cells were resuspended (4$\times$10$^6$ cells/ml) in RPMI 1640 followed by treatment with various concentrations of LPS (0, 0.1, 1, 10$\mu\textrm{g}$/ml) for 24 hours at 37$^{\circ}C$ in 5% $CO_2$ incubator. The supernatants of cells were collected and the levels of IL-1$\alpha$, IL=1$\beta$ and TNF-$\alpha$ were measured by enzyme-linked immunosorbent assay. The results were as follows ; 1. The levels of IL-1$\alpha$, IL-1$\beta$, TNF-$\alpha$ from PMN treated with each LPS were significantly higher than those released from unstimulated PMN of the control group (p<0.05). 2. The levels of all three cytokines released from PMN stimulated with each calcium hydroxide treated LPS were significantly lower than those released from PMN stimulated with each untreated LPS (p<0.05), while they were not significantly different from those released from unstimulated PMN of the control group (p>0.05) 3. The levels of secretion for all three cytokines were affected in a dose-dependent manner in PMN stimulated with each LPS (p<0.05), but not in PMN stimulated with each calcium hydroxide treated LPS (p>0.05). 4. The levels of all three cytokines released from PMN stimulated with p. endodontalis LPS were significantly lower than those released from PMN stimulated with E coli LPS (p<0.05).

• Korean Journal of Veterinary Research
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• v.29 no.2
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• pp.33-39
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• 1989

Pure separation of various leukocytes is required for the assessment of their roles in immunological and phisiological function. In this study, pure separation of monocytes from canine peripheral blood was attempted. At first, mononuclear cells (PBMC) were separated by ficoll-hypaque gradient method and then monocytes were recovered from PBMC suspensions in sucrose gradient Sol. (PBMC-Sucrose), autologous plasma (PBMC-Plasma) and autologous serum (PBMC-Serum) incubated at $37^{\circ}C$ for 2 hours. 1. In the separation of PBMC by ficoll-hypaque gradient method in canine blood, higher relative centrifugal force (RCF) was required, as high as more than 1,300xg RCF for 40 minutes, for clear formation of PBMC layer than that in human blood as usually used 400xg RCF for 40 minutes. 2. In monocytes-separation from three PBMC suspensions following PBMC separation, recovery-, purity- and viability-rate of monocytes showed better results in PBMC-Plasma and PBMC-Serum than in PBMC-Sucrose suspension, particulary showing better results from PBMC suspensions performed by centrifugation at 1,500xg RCF for 40 minutes.

• Korean Journal of Environmental Biology
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• v.23 no.4
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• pp.379-382
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• 2005

The new cationic meso-substituted N-quarternized 4-pyridylporphyrins and their metal derivatives were synthesized as novel chemotherapeutics. The level of DNA damage induced by porphyrins TOBut4PyP, TOBut4PyP, TOEt4PyPMn and TOBut4PyPMn and its dependence on the chemical structure of compounds were analyzed by the Comet-assay. On the base of data obtained, the investigated porphyrins may be arranged by their genotoxic activity in the following order: TOEt4PyP>TOEt4PyPMn>TOBut4PyP>TOBut4PyPMn. Thus, i) the genotoxicity of the Mn-derivatives of TOEt4PyP and TOBut4PyP is higher than the original porphyrins and ii) the genotoxicity of TOEt4PyP and TOEt4PyPMn is increased after substitution of a butyl radical for ethyl one. The applied Comet-assay permits to reveal the dependence of DNA damage induction on the chemical structure of porphyrins.

• The Journal of the Society of Korean Medicine Diagnostics
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• v.15 no.1
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• pp.47-54
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• 2011

In traditional Korean medicine, inspection of the tongue is an important method of making medical diagnoses and determining prognosis. We surveyed the fluorescence characteristics of the tongue coat in the ultraviolet light. The tongue coat comprises micro-organisms, blood metabolites, leukocytes from periodontal pockets, large amounts of desquamated epithelial cells released from the oral mucosa and different nutrients. In the ultraviolet light tissues of the oral cavity generally emit weak red or green fluorescence, which is not easily seen by the human eye, but is readily detected. This fluorescence has been proved to be due to the production of porphyrins by oral micro-organisms. While the composition of motile micro-organisms on the dorsum of the tongue is not constant, variations also occur persistingly in the fluorescence characteristics of the tongue coat. But because live bacteria contain a variety of intracellular biomolecules that have specific excitation and emission wavelength spectra characterizing their intrinsic fluorescence, the tongue coat emits fluorescence. the tongue itself, on the other hand, emits very weak or not fluorescence. In conclusion, we suggests that the uncoated tongue area be eliminated from the coated tongue area with the difference between the fluorescence characteristics of the tongue and that of the tongue coat.

• Archives of Pharmacal Research
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• v.28 no.11
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• pp.1257-1262
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• 2005

MMP-9 is a metalloproteinase capable of basement membrane degradation in vivo. Expression of MMP-9 can be found in normal conditions such as trophoblasts, osteoclasts, and leukocytes and their precursors. They also occur as well as in pathological conditions, such as the invasive growth of primary tumors, metastasis, angiogenesis, rheumatoid arthritis, and periodontal diseases. MMP-9 upregulation can be highly induced by a wide range of agents. These agents include growth factors, cytokines, cell-cell, and cell-ECM adhesion molecules, and agents altering cell shape. Here, we observed that TNF-$\alpha$ stimulated human monocytic cell line, HL-60 produced MMP-9 in a dose and time dependent manner. Real time PCR results indicated transcriptional upregulation of MMP-9 as early as 3 h post TNF-$\alpha$ stimulation. To investigate the signaling pathway underlined in TNF-$\alpha$ induced MMP-9 expression, three MAP kinase inhibitors were added to cells 1 h prior to TNF-$\alpha$ treatment. The ERK inhibitor completely abolished MMP-9 expression by TNF-$\alpha$. But neither p38 MAP kinase nor JNK inhibitor had an effect on TNF-$\alpha$ induced MMP-9 expression, suggesting that ERK activation is required for the MMP-9 induction by TNF-$\alpha$. Taken together, we found that TNF-$\alpha$ stimulation facilitates ERK activation, which results in the transcriptional upregulation of MMP-9 gene and subsequent MMP-9 production and secretion.

• Bulletin of the Korean Chemical Society
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• v.28 no.6
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• pp.1005-1009
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• 2007

Leukotriene B4 (LTB4) is a potent chemoattractant for leukocytes and considered to be an inflammatory mediator. Human BLT2 (hBLT2) is a low-affinity G-protein coupled receptor for LTB4 and mediates pertussis toxin-sensitive chemotactic cell movement. Here, we dissected the interaction between hBLT2 and G-protein alpha subunits using GST fusion proteins containing intracellular regions of hBLT2 and various Gα protein including Gα i1, Gα i2, Gα i3, Gα s1, Gα o1, and Gα z. Among the tested Gα subunits, Gα i3 showed the highest binding to the third intracellular loop region of hBLT2 with a dissociation constant (KD) of 5.0 × 10?6 M. These results suggest that Gα i3 has the highest affinity to hBLT2, and the third intracellular loop region of hBLT2 is the major component for the interaction with Gα i3.