• The Journal of Natural Sciences
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• v.4
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• pp.103-142
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• 1991

These studies were carried out to select somatic hybrid using selectable marker genes of Nicotiana glauca transformed by NPTII gene and Solanum tuberosum transformed by T- DNA, and to study characteristics of transformant. The results are summarized as follows. 1. Crown gall tumors and hairy roots were formed on potato tuber disc infected by A. tumefaciens Ach5 and A. rhizogenes ATCC15834. These tumors and roots could be grown on the phytohormone free media. 2. Callus formation from hairy root was prompted on the medium containing 2, 4 D 2mg/I with casein hydrolysate lg/l. 3. The survival ratio of crown gall tumor callus derived from potato increased on the medium containing the activated charcoal 0. 5-2. 0mg/I because of the preventions on the other hand, hairy roots were necrosis on the same medium. 4. Callus derived from hairy root were excellently grown for a short time by suspension culture on liquid medium containing 2, 4-D 2mg/I and casein hydrolysate lg/l. 5. The binary vector pGA643 was mobilized from E. coli MC1000 into wild type Agrobacteriurn tumefaciens Ach5, A. tumefaciens $A_4T$ and disarmed A. tuniefaciens LBA4404 using a triparental mating method with E. ccli HB1O1/pRK2013. Transconjugants were obtained on the minimal media containing tetracycline and kanamycin. pGA643 vectors were confirmed by electrophoresis on 0.7% agarose gel. 6. Kanamycin resistant calli were selected on the media supplemented with 2, 4-D 0.5mg/1 and kanamycin $100\mug$/ml after co- cultivating with tobacco stem explants and A. tumefaciens LBA4404/pGA643, and selected calli propagated on the same medium. 7. The multiple shoots were regenerated from kanamycin resistant calli on the MS medium containing BA 2mg/l. 8. Leaf segments of transformed shoot were able to grow vigorusly on the medium supplemented with high concentration of kanamycin $1000\mug$/ml. 9. Kanamycin resistant shoots were rooting and elongated on medium containing kanamycin $100\mug$/ml, but normal shoot were not. 10. For the production of protoplast from potato calli transformed by T-DNA and mesophyll tissue transformed by NPTII gene, the former was isolated in the enzyme mixture of 2.0% celluase Onozuka R-10, 1.0% dricelase, 1.0% macerozyme. and 0.5M mannitol, the latter was isolated in the enzyme mixture 1.0% Celluase Onozuka R-10, 0.3% macerozyme, and 0.7M mannitol. 11. The optimal concentrationn of mannitol in the enzyme mixture for high protoplast yield was 0.8M at both transformed tobacco mesophyll and potato callus. The viabilities of protoplast were shown above 90%, respectively. 12. Both tobacco mesophyll and potato callus protoplasts were fused by using PEG solution. Cell walls were regenerated on hormone free media supplemented with kanamycin after 5 days, and colonies were observed after 4 weeks culture.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.15 no.2
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• pp.271-288
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• 2002

Authors investigated the pathogenesis and treatment of dennatosurgical diseases in the ImJeungJiNamUiAn(臨證指南醫案). 1. The symptoms and diseases of dermatosurgery were as follows; 1) BanSaJinRa(반사진라) : eczema, atopic dermatitis, seborrheic dermatitis, psoriasis, lichen planus, pityriasis rosea, hives, dermographism, angioedema, cholinergic urticaria, urticaria pigmentosa, acne, milium, syringoma, keratosis pilaris, discoid lupus erythematosus, hypersensitivity vasculitis, drug eruption, polymorphic light eruption, rheumatic fever, juvenile rheumatoid arthritis(Still's disease), acute febrile neutrophilic dermatosis(Sweet's syndrome), Paget's disease, folliculitis, viral exanthems, molluscum contagiosum, tinea, tinea versicolor, lymphoma, lymphadenitis, lymphangitis, granuloma annulare, cherry angioma 2) ChangYang(瘡瘍) : acute stage eczema, seborrheic dermatitis, stasis ulcer, intertrigo, xerosis, psoriasis, lichen planus, ichthyosis, pityriasis rosea, rosacea, acne, keratosis pilaris, dyshidrosis, dermatitis herpetiformis, herpes gestationis, bullae in diabetics, pemphigus, lupus erythematosus, fixed drug eruption, erythema multiforme, toxic epidermal necrolysis, toxic shock syndrome, staphylococcal scaled skin syndrome, scarlet fever, folliculitis, impetigo, pyoderma gangrenosum, tinea, candidiasis, scabies, herpes simplex, herpes zoster, chicken pox, Kawasaki syndrome, lipoma, goiter, thyroid nodule, thyroiditis, hyperthyroidism, thyroid cancer, benign breast disorder, breast carcinoma, hepatic abscess, appendicitis, hemorrhoid 3) Yeok(疫) : scarlet fever, chicken pox, measles, rubella, exanthem subitum, erythema infectiosum, Epstein-Barr virus infection, cytomegalovirus infection, hand-foot-mouth disease, Kawasaki disease 4) Han(汗) : hyperhidrosis 2. The pathogenesis and treatment of dermatosurgery were as follows; 1) When the pathogenesis of BalSa(발사), BalJin(發疹), BalLa(발라) and HangJong(項腫) are wind-warm(風溫), exogenous cold with endogenous heat(外寒內熱), wind-damp(風濕), the treatment of evaporation(解表) with Menthae Herba(薄荷), Arctii Fructus(牛蒡子), Forsythiae Fructus(連翹) Mori Cortex(桑白皮), Fritillariae Cirrhosae Bulbus(貝母), Armeniaoae Amarum Semen(杏仁), Ephedrae Herba(麻黃), Cinnamomi Ramulus(桂枝), Curcumae Longae Rhizoma(薑黃), etc can be applied. 2) When the pathogenesis of BuYang(부양), ChangI(瘡痍) and ChangJilGaeSeon(瘡疾疥癬) are wind-heat(風熱), blood fever with wind transformation(血熱風動), wind-damp(風濕), the treatment of wind-dispelling(疏風) with Arctii Fructus(牛蒡子), Schizonepetae Herba(荊芥), Ledebouriellae Radix(防風), Dictamni Radicis Cortex(白鮮皮), Bombyx Batrytioatus(白？?), etc can be applied. 3) When the pathogenesis of SaHuHaeSu(사후해수), SaJin(사진), BalJin(發疹), EunJin(은진) and BuYang(부양) are wind-heat(風熱), exogenous cold with endogenous heat(外寒內熱), exogenous warm pathogen with endogenous damp-heat(溫邪外感 濕熱內蘊), warm pathogen's penetration(溫邪內陷), insidious heat's penetration of pericardium(伏熱入包絡), the treatment of Ki-cooling(淸氣) with TongSeongHwan(通聖丸), Praeparatum(豆？), Phyllostachys Folium(竹葉), Mori Cortex(桑白皮), Tetrapanacis Medulla(通草), etc can be applied. 4) When the pathogenesis of JeokBan(적반), BalLa(발라), GuChang(久瘡), GyeolHaek(結核), DamHaek(痰核), Yeong(？), YuJu(流注), Breast Diseases(乳房疾患) and DoHan(盜汗) are stagnancy's injury of Ki and blood(鬱傷氣血), gallbladder fire with stomach damp(膽火胃濕), deficiency of Yin in stomach with Kwolum's check (胃陰虛 厥陰乘), heat's penetration of blood collaterals with disharmony of liver and stomach(熱入血絡 肝胃不和), insidious pathogen in Kwolum(邪伏厥陰), the treatment of mediation(和解) with Prunellae Spica(夏枯草), Chrysanthemi Flos(菊花), Mori Folium (桑葉), Bupleuri Radix(柴胡), Coptidis Rhizoma(黃連), Scutellariae Radix(黃芩), Gardeniae Fructus(梔子), Cyperi Rhizoma(香附子), Toosendan Fructus(川？子), Curcumae Radix(鬱金), Moutan Cortex(牧丹皮), Paeoniae Radix Rubra(赤芍藥), Unoariae Ramulus Et Uncus(釣鉤藤), Cinnamorni Ramulus(桂枝), Paeoniae Radix Alba(白芍藥), Polygoni Multiflori Radix (何首烏), Cannabis Fructus (胡麻子), Ostreae Concha(牡蠣), Zizyphi Spinosae Semen(酸棗仁), Pinelliae Rhizoma(半夏), Poria(백복령). etc can be applied. 5) When the pathogenesis of BanJin(반진), BalLa(발라), ChangI(瘡痍), NamgChang(膿瘡). ChangJilGaeSeon(瘡疾疥癬), ChangYang(瘡瘍), SeoYang(署瘍), NongYang(膿瘍) and GweYang(潰瘍) are wind-damp(風濕), summer heat-damp(暑濕), damp-warm(濕溫), downward flow of damp-heat(濕熱下垂), damp-heat with phlegm transformation(濕熱化痰), gallbladder fire with stomach damp(膽火胃濕), overdose of cold herbs(寒凉之樂 過服), the treatment of damp-resolving(化濕) with Pinelliae Rhizoma(半夏), armeniacae Amarum Semen(杏仁), Arecae Pericarpium(大腹皮), Poria(백복령), Coicis Semen(薏苡仁), Talcum(滑石), Glauberitum(寒水石), Dioscoreae Tokoro Rhizoma(？？), Alismatis Rhizoma(澤瀉), Phellodendri Cortex(黃柏), Phaseoli Radiati Semen(？豆皮), Bombycis Excrementum(?沙), Bombyx Batryticatus(白？?), Stephaniae Tetrandrae Radix(防己), etc can be applied. 6) When the pathogenesis of ChangPo(瘡泡), hepatic abscess(肝癰) and appendicitis(腸癰) are food poisoning(食物中毒), Ki obstruction & blood stasis in the interior(기비혈어재과), damp-heat stagnation with six Bu organs suspension(濕熱結聚 六腑不通), the treatment of purgation(通下) with DaeHwangMokDanPiTang(大黃牧丹皮湯), Manitis Squama(穿山甲), Curcumae Radix(鬱金), Curcumae Longae Rhizoma(薑黃), Tetrapanacis Medulla(通草), etc can be applied. 7) When the pathogenesis of JeokBan(적반), BanJin(반진), EunJin(은진). BuYang(부양), ChangI(瘡痍), ChangPo(瘡泡), GuChang(久瘡), NongYang(膿瘍), GweYang(潰瘍), Jeong(정), Jeol(癤), YeokRyeo(疫？) and YeokRyeolpDan(疫？入？) are wind-heat stagnation(風熱久未解), blood fever in Yangmyong(陽明血熱), blood fever with transformation(血熱風動), heat's penetration of blood collaterals(熱入血絡). fever in blood(血分有熱), insidious heat in triple energizer(三焦伏熱), pathogen's penetration of pericardium(心包受邪), deficiency of Yong(營虛), epidemic pathogen(感受穢濁), the treatment of Yong & blood-cooling(淸營凉血) with SeoGakJiHwangTang(犀角地黃湯), Scrophulariae Radix(玄參), Salviae Miltiorrhizae Radix(丹參), Angelicae Gigantis Radix(當歸), Polygoni Multiflori Radix(何首烏), Cannabis Fructus(胡麻子), Biotae Semen(柏子仁), Liriopis Tuber(麥門冬), Phaseoli Semen(赤豆皮), Forsythiae Fructus(連翹), SaJin(사진), YangDok(瘍毒) and YeokRyeoIpDan(역려입단) are insidious heat's penetration of pericardium(伏熱入包絡), damp-warm's penetration of blood collaterals(濕溫入血絡), epidemic pathogen's penetration of pericardium(심포감수역려), the treatment of resuscitation(開竅) with JiBoDan(至寶丹), UHwangHwan(牛黃丸), Forsythiae Fructus(連翹), Curcumae Radix(鬱金), Tetrapanacis Medulla(通草), Acori Graminei Rhizoma(石菖蒲), etc can be applied. 9) When the pathogenesis of SaHuSinTong(사후신통), SaHuYeolBuJi(사후열부지), ChangI(瘡痍), YangSon(瘍損) and DoHan(盜汗) are deficiency of Yin in Yangmyong stomach(陽明胃陰虛), deficiency of Yin(陰虛), the treatment of Yin-replenishing(滋陰) with MaekMunDongTang(麥門冬湯), GyeongOkGo(瓊玉膏), Schizandrae Fructus(五味子), Adenophorae Radix(沙參), Lycii Radicis Cortex (地骨皮), Polygonati Odorati Rhizoma(玉竹), Dindrobii Herba(石斛), Paeoniae Radix Alba(白芍藥), Ligustri Lucidi Fructus (女貞子), etc can be applied. 10) When the pathogenesis of RuYang(漏瘍) is endogenous wind in Yang collaterals(陽絡內風), the treatment of endogenous wind-calming(息風) with Mume Fructus(烏梅), Paeoniae Radix Alba (白芍藥), etc be applied. 11) When the pathogenesis of GuChang(久瘡), GweYang(潰瘍), RuYang(漏瘍), ChiChang(痔瘡), JaHan(自汗) and OSimHan(五心汗) are consumption of stomach(胃損), consumption of Ki & blood(氣血耗盡), overexertion of heart vitality(勞傷心神), deficiency of Yong(營虛), deficiency of Wi(衛虛), deficiency of Yang(陽虛), the treatment of Yang-restoring & exhaustion-arresting(回陽固脫) with RijungTang(理中湯), jinMuTang(眞武湯), SaengMaekSaGunjaTang(生脈四君子湯), Astragali Radix (황기), Ledebouriellae Radix(防風), Cinnamomi Ramulus(桂枝), Angelicae Gigantis Radix(當歸), Ostreae Concha(牡蠣), Zanthoxyli Fructus(川椒), Cuscutae Semen(兎絲子), etc can be applied.

• The Korean Journal of Pharmacology
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• v.16 no.2 s.27
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• pp.55-70
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• 1980

The pharmacological and microbiological studies of Cefoperazone (T-1551, Toyama Chemical Co., Japan) were conducted in vitro and in vivo. The studies included stability and physicochemical characteristics, antimicrobial activity, animal and human pharmacokinetics, animal pharmacodynamics and safety evaluation of Cefoperazone sodium for injection. 1) Stability and physicochemical characteristics. Sodium salt of cefoperazone for injection had a general appearance of white crystalline powder which contained 0.5% water, and of which melting point was $187.2^{\circ}C$. The pH's of 10% and 25% aqueous solutions were 5.03 ana 5.16 at $25^{\circ}C$. The preparations of cefoperazone did not contain any pyrogenic substances and did not liberate histamine in cats. The drug was highly compatible with common infusion solutions including 5% Dextrose solution and no significant potency decrease was observed in 5 hours after mixing. Powdered cefoperazone sodium contained in hermetically sealed and ligt-shielded container was highly stable at $4^circ}C{\sim}37^{\circ}C$ for 12 weeks. When stored at $4^{\circ}C$ the potency was retained almost completely for up to one year. 2) Antimicrobial activity against clinical isolates. Among the 230 clinical isolates included, Salmonella typhi was the most susceptible to cefoperazone, with 100% inhibition at MIC of ${\leq}0.5{\mu}g/ml$. Cefoperazone was also highly active against Streptococcus pyogenes(group A), Kletsiella pneumoniae, Staphylococcus aureus and Shigella flexneri, with 100% inhibition at $16{\mu}g/ml$ or less. More than 80% of Escherichia coli, Enterobacter aerogenes and Salmonella paratyphi was inhibited at ${\leq}16{\mu}/ml$, while Enterobacter cloaceae, Serratia marcescens and Pseudomonas aerogenosa were somewhat less sensitive to cefoperagone, with inhibitions of 60%, 55% and 35% respectively at the same MIC. 3) Animal pharmacokinetics Serum concentration, organ distritution and excretion of cefoperazone in rats were observed after single intramuscular injections at doses of 20 mg/kg and 50 mg/kg. The extent of protein binding to human plasma protein was also measured in vitro br equilibrium dialysis method. The mean Peak serum concentrations of $7.4{\mu}g/ml$ and $16.4{\mu}/ml$ were obtained at 30 min. after administration of cefoperazone at doses of 20 mg/kg and 50 mg/kg respectively. The tissue concentrations of cefoperazone measured at 30 and 60 min. were highest in kidney. And the concentrations of the drug in kidney, liver and small intestine were much higher than in blood. Urinary and fecal excretion over 24 hours after injetcion ranged form 12.5% to 15.0% in urine and from 19.6% to 25.0% in feces, indicating that the gastrointestinal system is more important than renal system for the excretion of cefoperazone. The extent of binding to human plasma protein measured by equilibrium dialysis was $76.3%{\sim}76.9%$, which was somewhat lower than the others utilizing centrifugal ultrafiltration method. 4) Animal pharmacodynamics Central nervous system : Effects of cefoperazone on the spontaneous movement and general behavioral patterns of rats, the pentobarbital sleeping time in mice and the body temperature in rabbits were observed. Single intraperitoneal injections at doses of $500{\sim}2,000mg/kg$ in rats did not affect the spontaneous movement ana the general behavioral patterns of the animal. Doses of $125{\sim}500mg/kg$ of cefoperazone injected intraperitonealy in mice neither increased nor decreased the pentobarbital-induced sleeping time. In rabbits the normal body temperature was maintained following the single intravenous injections of $125{\sim}2,000mg/kg$ dose. Respiratory and circulatory system: Respiration rate, blood pressure, heart rate and ECG of anesthetized rabbits were monitored for 3 hours following single intravenous injections of cefoperazone at doses of $125{\sim}2,000mg/kg$. The respiration rate decreased by $3{\sim}l7%$ at all the doses of cefoperazone administered. Blood pressure did not show any changes but slight decrease from 130/113 to 125/107 by the highest dose(2,000 mg/kg) injected in this experiment. The dosages of 1,000 and 2,000 mg/kg seemed to slightly decrease the heart rate, but it was not significantly different from the normal control. All the doses of cefoperazone injected were not associated with any abnormal changes in ECG findings throughout the monitering period. Autonomic nervous system and smooth muscle: Effects of cefoperazone on the automatic movement of rabbit isolated small intestine, large intestine, stomach and uterus were observed in vitro. The autonomic movement and tonus of intestinal smooth muscle increased at dose of $40{\mu}g/ml$ in small intestine and at 0.4 mg/ml in large intestine. However, in stomach and uterine smooth muscle the autonomic movement was slightly increased by the much higher doses of 5-10 mg/ml. Blood: In vitro osmotic fragility of rabbit RBC suspension was not affected by cefoperazone of $1{\sim}10mg/ml$. Doses of 7.5 and 10 mg/ml were associated with 11.8% and 15.3% prolongation of whole blood coagulation time. Liver and kidney function: When measured at 3 hours after single intravenous injections of cefoperaonze in rabbits, the values of serum GOT, GPT, Bilirubin, TTT, BUN and creatine were not significantly different from the normal control. 5) Safety evaluation Acute toxicity: The acute toxicity of cefoperazone was studied following intraperitoneal and intravenous injections to mice(A strain, 4 week old) and rats(Sprague-Dawler, 6 week old). The LD_(50)'s of intraperitonealy injected cefoperazone were 9.7g/kg in male mice, 9.6g/kg in female mice and over 15g/kg in both male and female rats. And when administered intravenously in rats, LD_(50)'s were 5.1g/kg in male and 5.0g/kg in female. Administrations of the high doses of the drug were associated with slight inhibition of spontaneous movement and convulsion. Atdominal transudate and intestinal hyperemia were observed in animals administered intraperitonealy. In rats receiving high doses of the drug intravenously rhinorrhea and pulmonary congestion and edema were also observed. Renal proximal tubular epithelial degeneration was found in animals dosing in high concentrations of cefoperazone. Subacute toxicity: Rats(Sprague-Dawley, 6 week old) dosing 0.5, 1.0 and 2.0 g/kg/day of cefoperazone intraperitonealy were observed for one month and sacrificed at 24 hours after the last dose. In animals with a high dose, slight inhibition of spontaneous movement was observed during the experimental period. Soft stool or diarrhea appeared at first or second week of the administration in rats receiving 2.0g/kg. Daily food consumption and weekly weight gain were similar to control during the administration. Urinalysis, blood chemistry and hematology after one month administration were not different from control either. Cecal enlargement, which is an expected effect of broad spectrum antibiotic altering the normal intestinal microbial flora, was observed. Intestinal or peritoneal congestion and peritonitis were found. These findings seemed to be attributed to the local irritation following prolonged intraperitoneal injections of hypertonic and acidic cefoperazone solution. Among the histopathologic findings renal proximal tubular epithelial degeneration was characteristic in rats receiving 1 and 2g/kg/day, which were 10 and 20 times higher than the maximal clinical dose (100 mg/kg) of the drug. 6) Human pharmacokinetics Serum concentrations and urinary excretion were determined following a single intravenous injection of 1g cefoperazone in eight healthy, male volunteers. Mean serum concentrations of 89.3, 61.3, 26.6, 12.3, 2.3, and $1.8{\mu}g/ml$ occured at 1,2,4,6,8 and 12 hours after injection respectively, and the biological half-life was 108 minutes. Urinary excretion over 24 hours after injection was up to 43.5% of administered dose.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.4 no.1
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• pp.37-53
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• 1998

The sprig of Jinryungtang Gagambang has been used for curing as a traditional medicine without any experimental evidence to support the rational basis for their clinical use. This experiment was carried out to evaluate the possible therapeutic or antitumoral effects of Jinryungtang Gagambang extract against cancer, and to study some mechanisms responsible for its effect. The cytotoxic and antitumor effects were evaluated on human cell liens (A549, hep3B, Caki-1, Sarcoma 180) after exposure to Jinryungtang Gagambang extract using in ILS, colony forming efficency and SRB assay which were regarded as a valuable method for cytotoxic and antitumor effects of unknown compound on tumor cell lines. The results obtained in this studies were as follows. 1. As a result of exposure to Jinryungtang Gagambang extract, the proliferation of A549, hep3B, Caki-1, good correlations were shown from the results of SRB assay and those of clogenetic assay. 2. The oral administration of Jinryungtang Gagambang extract showed significant effects of increase of MST(mean survival time) and ILS(increased life span) depending on the increasing concentration. 3. Against squamous cell carcinoma induced by MCA, Jinryungtang Gagambang decreased not only the frequency of tumor production but also the number and weight of tumors per tumor bearing mice(TBM). Jinryungtang Gagambang also significantly suppressed the development of 3LL cell-implanted tumors by frequency and their size, and some developed tumors were regressed by the continuous treatment of Jinryungtang Gagambang extract into TBM. 4. Jinryungtang Gagambang extract also increased NK cell activities. According to the above results, it could be suggested that Jinryungtang Gagambang extract has prominent antiutmor effect.