염증반응이란 우리 신체에 외부 물질이 침입했을 경우 대처하는 정상적인 현상이다. 외부에서 물리, 화학, 생물학적 물질에 노출되었을 경우 혈관이 팽창하고, 비특이적 면역반응이 시작된다. Selectin, integrin, immunoglobulin superfamily와 같은 adhesion molecule에 의해 macrophage 및 neutrophil과 같은 leukocyte의 반응을 촉진하기 위해 exudation이 일어나고 침입 부위에 모여 hyperemia가 발생된다.(Govan 등, 1995)

This study was performed to examine the anti-cancer effect of Red Ginseng in the DENGalN-PH-induced hepatic tumor model system in rats. One hundred of male SPF Sprague-Dawley rats(6weeks old) were randomly divided into five groups. Rats in groups 1, 2, 3, and 4 were administered to diethylnitrosamine intraperitoneally 200 mg/kg body weight for the caner initiation. Rats in group 5 were given to saline as a control. On two weeks after cancer initiation, rats in groups 1 and 3 were fed on diet containing 0.01% of acethylaminofiuorene(AAF) which is strong cancer-promotor for 6 weeks, while rats in groups 2 and 4 were fed on water containing 0.05% of phenobarbital which is weak cancer.promotor for 6 weeks. Rats in groups 1 and 2 were treated with diet containing 3% of Red Ginseng for six weeks(from 9th week till 15th week after cancer initiation). Rats in all groups were necropsied time-sequencially at 8, 15, and 36 weeks. The hepatic lesions of rat treated with carcinogens expressed glutathione S-transferase placental form(GST-P) at 8 week. The GST-P positive foci of rats treated with AAF were larger than that of any other rats, while the GST-P positive foci of rats treated with AAF and red ginseng were significantly decreased. This anti-cancer effect of Red ginseng might be involved in the enhacement of natural killer cell activity. To know whether there is direct relationship between Red Ginseng and natural killer cell activity, the activity of natural killer cell was examined after treatment AAF, AAF+Red ginseng and Red ginseng only, respectively. Comparing with natural killer cell activity in AAF-treated group, natural killer cell activity was significantly activated in AAF+ Red ginseng-treated group. This indicated that Red ginseng might enhance natural killer activity after treatment carcinogen in rats. These results suggested that Red ginseng might have a cancer prevention ability by promoting natural killer cell activity during hepatocarclnogenesis.

Cyclophosphamide(25, 50 or 100 mg/kg), orally administered to male Sprague-Dawley rats, caused a time- and dose-dependent thymic atrophy. In the light microscopic examination of the atrophic thymus, thymocytes with condensed or fragmented nucleus were multifocally observed in the cortical region, started to increase 8 hr after CPA treatment and reached to the maximal level at 16 hr, although such cells were not seen after 48 hr when the severe depletion of thymocytes were marked. In agarose gel electrophoresis to analyze the DNA changes, DNA extracted from atrophic thymus showed a oligonucleosomal laddering at the corresponding time to morphological changes. In an additional supportive experiment, thymocytes showing morphological changes, nuclear condensation or apoptotic body, exhibited a positive reaction to immunoperoxidase staining using in situ apoptosis detection kit. Separately, agarose gel electrophoresis of DNA from bone .marrow cells was performed to investigate the involvement of bone marrow cells in the process of thymocyte apoptosis. Although DNA laddering was slightly increased 2 and 4 hr after treatment, no clear correlation was inferred. Taken togather, it is concluded that thymocytes showing morphological changes in thymic atrophy induced by cyclophosphamide administration represent an apoptosis having biochemical nature of programmed cell death.

PCBs are classified as B2 (Probable human carcinogen) based on the induction of hepatocellular carcinomas in rats and mice from IRIS (Integrated Risk Information System). About 20 years ago, PCBs were phased out for electrical use in Korea, but PCBs were continuously used in the other field. Lately, there has been increasing concern on possible effects of contaminated soil to the other environment and human health. The purpose of this study is to determine PCBs level in soil at some site and to assess the human exposure doses according to exposure routes for people living within sites which expected to be exposed to PCBs. Pollution level of PCBs on the site was monitored using gas liquid chromatography. To assess the transport of PCBs in soil to plant and to air, various transfer factors(diffusion coefficient, bioconcentration factor etc.) were considered in simple calculations. To calculate the residential exposure doses by routes, some equations were considered using assumption value, which define inhalation, ingestion (soil, plant) and derreal uptake pathway. Computated results will be used as risk assessment information for human health evaluation on contaminated soil.

Gamma irradiation was applied to Angelicae gigantis radix and Aloe to improve their hygienic quality. The effective dose of irradiation was 7 kGy in Angelicae gigantis radlx and 5 kGy in Aloe for the sterilization of all contaminated microorganisms tested. After 8 months of storage at room temperature, no growth of microorganisms was observed in the irradiated products. The safety of these products were evaluated by Salmonella typhimurium reversion assay and in vivo micronucleus assay using mouse bone marrow cells. They were negative in the bacterial reversion assay with S. typhimurium TA 98, TA100, TA1535 and TA1537. In the in vivo mouse micronucleus assay, they did not show any clastogenic effect at all doses tested. These results indicate that the gamma irradiation of Angelicae gigantis radix at 12 kGy and of Aloe at 10 kGy have no genotoxic effects under these experimental conditions.

Exposure of splenocytes to 2-chloroethylethyl sulfide (CEES) resulted in the cell death, and the cytotoxicity of CEES was prevented by inhibitors of lysosomal hydrolases. Therefore, it has been postulated that the cytotoxicity of CEES may be partially due to the lysosomal labilization. This study, based on this mechanism, was undertaken to determine whether aminoglycoside antibiotics as inhibitors of lysosomal phospholipases and their combinations with other lysosome stabilizers can be useful as a treatment to reduce the CEES toxicity in mice. 2-Chloroethylethyl sulfide (20 mg/kg body weight) was injected ip into female ICR mice, and candidate compounds were administered ip before or after the CEES challenge. Kanamycin (40 mg/kg body weight) as effective as deferoxamine (100 mg/kg body weight) enhanced the survival rate after 5 days of intoxication from 10% of control to 50 - 60%. The most effective was found to be the combination of kanamycin, cycloheximide, deferoxamine and dextrose showing an almost full protection against 2LD50 of CEES. Consistent with the protection of the CEES toxicity, the decrease of body weight in mice intoxicated with CEES was effectively prevented by kanamycin or its combinations. It is suggested that kanamycin or its combination (kanamycin, cycloheximide, deferoxamine and dextrose) would be one of effective antidotes against the CEES poisoning in mice.

The mutational effects of pentachlorophenol (PCP) on the hypoxanthine phosphoribosyl transf erase (hprt) locus in human T-cell were analysed by T-cell clonal assay in vitro. Cells were exposed for 24 hours at primary culture to 0~100 ppm (W/V) PCP in dimethyl sulfoxide. Treated cells were allowed at the same time to stimulate by phytohemagglutinin (PHA) and T-cell growth factor (TCGF) and then seeded in medium containing 6-thioguanine to select for hprt-negative routants. We have also defined the optimal condition for the determination of mutant frequency. The parameters investigated include survival counting, first and second subculture for clonal efficiency plating and mutant plating. Under the optimal conditions, mutant frequencies of high dose-treated cells were significantly higher than those of non-treated or low dose cells. The results indicated a clear dose-effect relationship and showed that mutant frequency in 50 ppm PCP treated cell was 4.31$\times$$10^{-5}$ (background, 8.32$\times$$10^{-6}$). Above data strongly suggest that hprt mutation assay can be used as a biomarker for the environmental risk assessment.

Inhibitory effects of tannic acid on skin toxicity and heat shock protein induced by UVB were investigated. Tannic acid was administered either topically or orally for 3 days to hairless mice, which were previously irradiated with UVB. UVB was found to cause skin erythema . However, the skin erythema was decreased when tannic acid was administered either topically or orally. The heat shock proteins, Hsp-78 kDa and 70 kDa, were induced by UVB irradiation, but the induction was decreased by treatment of tannic acid in both topically and orally administered groups. The hsp induction was more prominent in orally administered groups than in topically administerd groups. However, the difference between two groups was not statistically significant. The route of administrations, topical and oral, does not affect the activity of tannic acid. In the skin tissue observation, tannic acid regenerated the epithelial cells with 7-9 cell layers which were injured by UVB. In conclusion, tannic acid has an ability to protect against UVB irradiation and regenerate the skin.

This study was carried out to investigate toxicity of insecticide carbofuran and compensatory effects of phenobarbital sodium (PB) in vivo and in vitro. Sprague Dawley male rats were used as experimental animals and divided into carbofuran only administered group and simultaneous application group of carbofuran and PB. At 30 rain and 1, 3, 6, 12, 24, 48 and 96 hrs after each treatment, the animals were sacrificed by decapitation. Kidney were immediately removed, immersed in fixatives, and processed with routine method for light microscopic study. Paraffin sections were stained with H-E, PAM and PAS. $5.0\times 10^4$ cell/ml of NIH 3T3 fibroblast in each well of 24 multidish were cultured: After 24 hours, the cells were treated with solution of six groups; control group cultured in media only, carbofuran $MTT_50$ or $NR_50$ group cultured in the media containing carbofuran $MTT_50$ or $NR_50$ and four experimental groups cultured in the media containing carbofuran $NR_50$ plus various concentratins of PB. After the NIH 3T3 fibroblast of all groups were cultured in same condition for 48 hours, Tetrazolium MTT (MTT) and NR (neutral red) assay were performed to evaluate the cytotoxicity of cell organelles. Under the light microscope, atrophic change of renal corpuscles were frequently observed in 1 and 2 days after carbofuran treatment. The increase of the mesangium was apparent in 1 and 2 days after carbofuran treatment. Necrotic changes of the epithelium and loss of brush border of proximal tubules were most severe at 2 and 3 days after carbofuran treatment, respectively. In contrast, there were no evidences of the toxic effects on renal tissues at 48hrs in carbofuran-PB treated groups. Carbofuran $MTT_50$ and $NR_50$ were 78$\mu M$, 82.5$\mu M$ respectively. MTT and NR quantities were significantly increased in carbofuran-PB 100$\mu M$ treatment group and carbofuran-PB 100$\mu M$ treatment group. On the basis of these results, it is obvious that PB has compensatory effects against carbofuran toxicity.

Ethyl carbamate (EC) is a potent teratogen in rodents and is present at low concentration in fermented foods and alcohol beverages. It has been well hypothesized that some metabolic products are responsible for the teratogenic effects of the compound. In the present study, the effects of phenobarbital (PB) on EC-induced embryotoxicity were investigated in SD rats. Six groups were constructed: EC 300 (EC 300 mg/kg/day), EC 600 (EC 600 mg/kg/day), EC 600+PB (EC 600 mg/kg/day and PB 80 mg/kg/day), PB (PB 80 mg/kg/day), DR (dietary restriction, 8 g/day/rat) and a control group. Rats of the EC 600+PB group were pretreated with phenobarbital intraperitoneally for three days to induce cytochrome P450 enzymes, followed by oral administration of EC for two consecutive days. The incidence of fetal deaths in the EC 600+PB group was higher than that of the EC 600 group(42.7 vs. 14.3%). The incidence of fetal realformations in the EC 600+PB group was higher than that of the EC 600 group (external; 7.0 vs. 4.1%, visceral; 31.4 vs. 11.3%, skeletal; 11.1 vs. 6.5%). There was no embryotoxicity in the control, EC 300, PB and DR groups. These results show that the pretreatment with phenobarbital augments EC-induced embryotoxicity in rats, indicating an evidence that metabolic activation by cytochrome P450 may be the major pathway of EC to its embryotoxic forms.

This study was performed to investigate the effects of cadmium chloride on the acute and chronic toxicity on rats. Several toxic effects of cadmium has been shown following short-term and longterm pretreatment with cadmium and zinc. Four groups of rats (A, B, C, D), each consisting of 16 rats, were studied and each group was divided into four subgroups (1, 2, 3, 4), 4 rats for each subgroup. Rats were subcutaneously pretreated with $CdCl_2$ (0.5 mg/kg, A & C), and $ZnCl_2$ (13.0 mg/kg, B & D) during time periods of 1 weeks (group A & B) and 6 weeks (group C & D). At the end of the period, rats were challenged with $CdCl_2$ (3.0, 6.0 and 9.0 mg/kg, i.p.). After giving the challenge dose, cadmium and metallothionein(MT) concentrations were determined. The concentrations of cadmium were higher in the liver than the kidney irrelevantly to cadmium and zinc pretreatment and increased dose-dependently to the challenge dosage. The metallothioneins showed higher concentrations in the liver than the kidney following cadmium pretreatment and were higher in the long-term pretreatment groups than the short-term pretreatment groups in the liver and the kidney of rats. These data suggest that metallothioneins are induced preferentially in the liver by pretreatment of cadmium and then, formed in the form of Cd-MT, may play an important role in the nephrotoxicity.

In the present study, we investigated the neutralization activity of various antimicrobials against lipopolysaccharide (LPS) as well as interaction between two antimicrobials (enrofioxacin and coilstin) using checkerboard method. The neutralization activity of antimicrobials used in the test was assayed by means of LAL chromogenic test after reaction of LPS with colistin, enorfioxacin, ampicillin, polymyxin B, oxytetracycline, streptomycin, and erythromycin. As the results, the neutralization activity of coltstin and polymixin B had a more stronger than that of tested other antimicrobials. In bacterial culture broth, the best neutralization activity of the antibiotics was also shown to coltstin and polymixin B. Meanwhile, It was shown to have synergism between enorfloxacin and coltstin on the basts of FIC (fractional inhibition concentration). The FIC of enorfioxacin-colistin combinations was 0.50-1.03 to Staphylococcus aureus R-209, 1.03-1.06 to Salmonella typhimurium, 0.75-1.25 to Bordetella Bronchtseptica and 1.02-1.25 to E. coli K88ab. On the basts of the above results, the present study may be of clinical usefulness in the choice of an antibiotic therapy for severe sepsts in animals.

In order to assess the possibility whether CYP2D is involved in caffeine metabolism, we have purified and characterized the rat liver microsomal cytochrome P4502D1 (CYP2D1), equivalent to CYP2D6 in human liver, and have utilized the reconstituted CYP2D1 in the metabolism of 4 primary caffeine (1, 3, 7-trimethylxanthine) metabolites such as paraxanthine (1, 7-dimethylxanthine), 1, 3, 7-trimethylurate, theophylline (1, 3-dimethylxanthine) and theobromine (3, 7-dimethylxanthine). Rat liver CYP 2D1 has been purified to a specific content of 8.98 nmole/mg protein (13.4fold purification, 1.5% yield) using $\omega$-aminooctylagarose, hydroxlapatite, and DE52 columns in a sequential manner. As judged from sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), the purified CYP2D1 was apparently homogeneous. Molecular weight of the purified CYP2D1 was found to be 51, 000 Da. Catalytic activity of the purified and then reconstituted CYP2D1 was confirmed by using bufuralol, a known subsFate of CYP2D1. The reconstituted CYP2D1 was found to produce to 1-hydroxylbufuralol at a rate of 1.43$\pm$0.13 nmol/min/nmol P450. The kinetic analysis of bufuralol hydroxylation indicated that Km and Vmax values were 7.32$\mu M$ and 1.64 nmol/min/nmol P450, respectively. The reconstituted CYP2D1 could catalyze the 7-demethylation of PX to 1-methylxanthine at a rate of 12.5 pmol/min/pmol, and also the 7- and 3- demethylations of 1, 3, 7-trimethylurate to 1, 3-dimethylurate and 1, 7-dimethylurate at 6.5 and 12.8 pmol/min/pmol CYP2D1, respectively. The reconstituted CYP2D1 could also 3-demethylate theophylline to 1-methylxanthine at 5 pmol/min/pmol and hydroxylate the theophylline to 1, 3-dimethylurate at 21.8 pmol/min/pmol CYP2D1. The reconstituted CYP2D1, however, did not metabolize TB at all (detection limits were 0.03 pmol/min/pmol). This study indicated that CYP2D1 is involved in 3-and 7-demethylations of paraxanthine and theophylline and suggested that CYP2D6 (equivalent to CYP2D1 in rat liver) present in human liver may be involved in the secondary metabolism of the primary metabolites of caffeine.

Acute toxicity of EPO(Erythropoietin) was investigated using rats and beagle dogs according to Established Regulation of Korean National Institute of Safety Research (1994. 4. 14). Rats and beagle dogs were injected intravenously with dosages of 20000 IU/kg, 2000 IU/kg, 200 IU/kg, 20 IU/kg and 2 IU/kg. In animals injected with EPO, there were neither dead animals nor significant changes of body weights. In addition, no differences were found between control and treated groups in clinical signs and autopsy findings. Therefore $LD_50$ of EPO was considered to be higher than 20000 IU/ kg B. W. in rats and beagle dogs.

Group of 40 male and 40 female Sprague-Dawley rats were given daily intravenous injections of different dosage of Erythropoietin (EPO), 80 IU/ kg/day (low dosage group), 400 IU/ kg/day (middle dosage group), or 2000 IU /kg/day (high dosage group)for 4 weeks by tail vein according to Established Regulation of Korean National Institute of Safety Research (1994. 4. 14). Appearance, behavior, mortality, and food consumption of rats of treated groups were not affected during the experimental periods. No significant EPO (erythropoietin)-related changes were found in urinalysts, eye examination, hematology, serum chemistry, and organ weight. No histopathological lesions were observed in both control and treatment groups. Our results strongly suggest that no toxic changes were found in rat treated intravenously with EPO (erythropoietin)for 4 weeks.

Four-week toxicity of EPO(erythropoietin) was investigated using New Zealand White rabbits according to the established regulations of Korean National Institute of Safety Research. Rabbits were administered intravenously seven days per week for 28 days with dosage of 0, 80, 400 and 2000IU/kg B. W./day. Animals administered with EPO showed no significant changes of body weight, water consumption and feed consumption, and no clinical signs and death. They were not significantly different from the control group in hematological and serum biochemical analysis, urinalysis, prothrombin time, and partial thromboplastin time. In this study, we concluded that EPO had no toxic effect in the New Zealand White rabbits when they were administered intravenously below 2000IU/ kg B.W./ day for 28 days.

This test was performed to evaluate the ocular and skin irritation of EPO (Erythropoietin). The results as follows: 1. Ocular irritation test There were no observed clinical signs, body weght changes by EPO during experimental period. The acute ocular irritation index(A.O.I.), mean ocular irritation index(M.O.I.) and Day-7 individual ocular irritation index(I.O.I.) of EPO at dose of 1000U and 10, 000U were 0, respectively. Therefore we evaluated that EPO was non-toxic to eyes. 2. Skin irritation test There were no observed clinical signs, body weght changes and gross pathologic findings by EPO during experimental period. There were no observed erythema, eschar formation and edema formation on intact and abraded skin treated by EPO. The primary irritation index(P.I.I.) of EPO at dose of 1000U and 10, 000U were 0, respectively and were evaluated none irritating product about skin irritation.

The antigenicity of EPO (erythropoietin) was investigated in guinea pig, mice and rats. Antigenicity tests-active systemic anaphylaxis (ASA), passive cutaneous anaphylaxis (PCA) of this materials were performed according to the established Regulation of Korean National Institute of Safety Research (1996, 4, 16). The results were followed: 1. After sensitizaion with EPO emulsified with complete Freund's adjuvant (CFA), guinea pigs didn't show any anaphylatic shock symptom in the ASA test 2. After sensitization with antisera of EPO sensitized mice, blue spots were observed on the hypodermis of back of rats in the PCA test, but diameter of each spot was smaller than 5 mm. From the results of this investigation, the antigenicity of EPO was negative under the present experimental condition.

The primary skin and eye irritancy of the combined vaccine (KGCC-95VI) for the prophylaxis against Japanese encephalitis and Hantaan virus infection recently developed by Korea Green Cross Corporation was investigated. The KGCC-95VI was applied to the back skins of the New Zealand White rabbits. The rabbits were observed for 72 hours and did not exhibit erythema, eschar and edema. The eyes of the rabbits were exposed to the KGCC-95VI. The rabbits were observed for 7 days and did not exhibit any ocular findings on cornea, iris and conjuntivae. The KGCC-95VI is considered not to have the primary skin and eye toxicity in rabbits.

This study was conducted to investigate the skin irritation by transdermal administration of the three dyes. These dyes were originated from onion by using extraction method. By the order of extraction from onion, A-dye was obtained from onion by using water at 90-100$\circ$C. B-dye was extracted from A-dye with ethylacetate. After ethylacetate extraction from A-dye, the lower layer named as C-dye. Twenty-four New Zealand white rabbits were divided into three groups. The each groups was consisted of two subgroups according to high dose (extracted dyes) and low dose (the 100-fold dilutions of A-, Band C-dye). In primary skin irritation test of male New Zealand White rabbits, body temperature and weights were not significantly changed and blood cells were positioned in normal blood cell ranges of health rabbits. Primary irritation index was "0" in the test and control sites of all animals used in this study. By the results obtained in the present test, all dyes were evaluated as a non-irritant on the basis of the criteria of Draize.of Draize.

LBD-001, a recombinant human interferon $\gamma$ produced by genetically engineered yeast as a host system, was intravenously administered to pregnant female rabbits (New Zealand White strain) from day 6 to 18 of gestation at dose levels of $0.35 \times 10^6$, $0. 69 \times 10^6$, and $1.38 \times 10^6$ I.U./kg/day. Hydrocortisone sodium succinate (0.3 mg/kg/day) was also given in the same way. Teratological effects of the test agents on the organogenesis of fetuses and the development of offsprings (F1 rabbits) were investigated. The results were as followings: (1) No significant changes by the treatment of LBD-001 or hydrocortisone sodium succinate were observed in the body weights, the food and water consumption, the lactating or nurshing behaviors, and the autopsy of the pregnant rabbits. (2) No significant changes in the resorption rate, the fetal organogenesis, and the normal develpoment of offsprings (F1) by the treatment of LBD-001 or hydrocortisone sodium succinate were detected. The results show that LBD-001 at the dose of $1.38 \times 10^6$ I.U./kg/day or less and hydrocortisone sodium succinate at the dose of 0.3 mg/kg/day are neither teratogenic in the organogensis of the fetuses and the development of the offsprings (F1) nor toxic to the mother rabbits.

LBD-001, a recombinant human interferon $\gamma$ produced by genetically engineered yeast as a host system, was intravenously administered to pregnant female rats (Sprague-Dawley) from day 17 of gestation to day 21 of lactation at dose levels.of $0.35 \times 10^6$, $0.69 \times 10^6$, and $1.38 \times 10^6$ I.U./kg/day. In vasopressin-treated group, vasopressin (5 I.U./kg/day) was intravenously injected only for 5 days of perinatal period. (1) No signicant changes by the treatment of LBD-001 were observed in the body weights, food and water consumption, feeding and nurshing behaviors, and the weights of main organs of mother rats. In vasopressin-treated group, no significant changes were observed except the decrease in the food consumption on day 18 of gestation and one case of abnormal offspring with bleeding spots on the skin. (2) No significant changes in the body weights, survival rates, locomotor activity, emotional development. and the motor coordination of offsprings (F1) by the treatment of LBD-001 were observed except the fact that increase of ambulation in the female offsprings of LBD-001 ($0.69 \times 10^6$ or $1.38 \times 10^6$ I.U./kg/day)-treated groups and the increase of rearing in the males of LBD-($1.38 \times 10^6$ I.U./kg/day)-treated group, and the increase of the weight of liver and ovaries in the female offsprings in the LBD-001 ($1.38 \times 10^6$ I.U./kg/day)-treated group were observed. Altogether, the results show that LBD-001 at the dose of $1.38 \times 10^6$ I.U./kg/day or less does not significantly affect the mother rats and their offsprings (F1) except the minor influences when treated during the perinatal and postnatal period.