• Research in Plant Disease
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• v.12 no.2
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• pp.139-143
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• 2006

Virion captured reverse transcription polymerase chain reaction (VC/RT-PCR) could detect plant virus quickly and accurately. In the VC/RT-PCR, no antibody is needed unlike immuno-captured RT-PCR (IC/RT-PCR) which had been improved method of RT-PCR for plant viruses, and virus nucleic acids can be obtained easily within 30minutes by property of polypropylene PCR tube which is hold and immobilized viral particles on its surface. For the virion capture of Tomato spotted wilt virus (TSWV), the extraction buffer was tested. The optimum macerating buffer for TSWV was 0.01M potassium phosphate buffer, pH 7.0, containing 0.5% sodium sulfite. The viral crude sap was incubated for 30 min at $4^{\circ}C$. The virions in the PCR tubes were washed two times with 0.01M PBS containing 0.05% Tween-20. The washed virions were treated at $95^{\circ}C$ immediately for 1 min containing RNase free water and chilled quickly in the ice. Disclosed virions' RNAs by heat treatment were used for RT-PCR. Dilution end point of $10^{-5}$ from plant's crude sap infected with TSWV showed relatively higher detection sensitivity for VC/RT-PCR. During multiple detection using two or more primers, interference was arisen by interactions between primer-primer and plant species. The result of multiplex RT-PCR was influenced by combinations of primers and the kind of plant, and the optimum extraction buffer for the multiplex detection by VC/RT-PCR should be developed.

• Research in Plant Disease
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• v.12 no.2
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• pp.91-98
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• 2006

A filtrate powder, designated as KNF2022, produced from culture broth of Acinetobacter sp. KTB3 was tested for their inhibitory effects on Pepper mild mottle virus (PMMoV) infection to Nicotiana glutinosa or N. tabacum cv. Xanthi nc. When 1/100 dilution with distilled water was treated to the plants and PMMoV was inoculated, the inhibition was estimated to be 94.3 and 95.6%, respectively. The same concentrations of KNF2022 inhibited infections of Pepper mottle virus (PepMoV) and Cucumber mosaic virus (CMV) on Chenopodium amaranticolor by 97.1 and 92.5%, respectively. Duration of inhibitory activity of the filtrate powder from Acinetobacter sp. culture broth against PMMoV infection on N. glutinosa was maintained for 2 days at 80% inhibition level, however, the inhibitory effect was diminished from 4 days after treatment to 50% levels. To evaluate inhibitory effects on systemic host plants of the antiviral agent, symptom developments of PMMoV, PepMoV and CMV on KNF2022-treated pepper plants were investigated. Delayed symptom developments until 10 days after inoculation (DAI) were observed for all the three viruses when the viruses were inoculated individually, and these delayed symptom development effects were maintained until 30 DAI in case of PepMoV. Moreover, PepMoV was not detected by RT-PCR and ELISA until 30 DAI. These delayed symptom development effects were diminished in all combinations of three virus co-inoculations due to synergism of three viruses on symptom developments. Inhibitory effect of KNF2022 was verified under electron microscopic examinations using purified virus preparations. Particles of PMMoV and PepMoV were observed on specimens from 5 min after KNF2022 treatment, and the particle sizes were reached in the range of 200-250 nm and 400-600 nm, respectively. Furthermore, the viral particles were destructed and particle sizes were reached in the range of 100-150 nm and 300-500 nm, respectively, on 60 min after treatments. Reduction of local lesion numbers on N. tabacum cv. Xanthi nc and C. amaranticolor were accompanied with reduction of virus particle sizes. In the case of CMV destructed particle numbers were also increased according to incubation period after KNF2022 treatment and local lesions on C. amaranticolor were reduced.

• Radiation Oncology Journal
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• v.19 no.3
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• pp.265-274
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• 2001

Purpose : Proctitis is one of acute complications encountered when radiotherapy was appled to the pelvis. Radiation-induced proctitis represents similar microscopic findings that are observed in inflammatory bowel disease (IBD). Nitric oxide (NO) plays an important role in the inflammatory process and many data suggest a close relationship between NO production and gastrointestinal inflammation. This study was aimed to establish the optimal radiation dose for radiation-induced proctitis in rat and to find a relationship between radiation proctitis and NO production. Materials and methods : Female Wistar rats, weighing from 150 to 220 g, received various doses(10-30 Gy) of radiation to the rectum. On the 5th and 10th day after irradiation, rectal specimens were evaluated grossly and microscopically. In addition, the degree of NO production by irradiation dose was evaluated by study with NOS expression and nitrite production in the irradiated rectal tissue. To evaluate relationship between radiation proctitis and NO, we administered aminoguanidine, iNOS inhibitor and L-arginine, substrate of NOS to rats from 2 days before to 7 days after the irradiation. Results : There were obvious gross and hostological changes after 17.5 Gy or higher radiation dose but not with 15 Gy or less radiation dose. Twenty Gy or higher dose of radiation caused Grade 4 damage in most of rectal specimens which were more likely to be related to the late complications such as fibrosis, rectal bleeding and rectal obstruction. A single fraction of 17.5 Gy to the rat rectum is considered to be an optimal dose to produce commonly experienced proctitis in the clinic. The result demonstrated that severity of microscopic damage of rectal mucosa from irradiation significantly correlated with iNOS over-expression. However, administration of iNOS inhibitor or substrate of iNOS did not influence the degree of rectal damage. Conclusion : A single fraction of 17.5 Gy irradiation to the rat rectum considered to be an optimal dose for radiation induced proctitis model. These results indicated that an excess production of NO contributes to pathogenesis of radiation-induced proctitis in part but was not the direct cause of rectal damage.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.1
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• pp.163-170
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• 2006

Nardostachyos Rhizoma (N. Rhizoma) belonging to the family Valerianaceae has been anti-arrhythmic effect, and sedation to the central nerve and a smooth muscle. We reported that the water extract of N. Rhizoma induced apoptotic cell death and differentiation in human promyelocytic leukemia (HL-60) cells. Cytotoxicity of N. Rhizoma was detected only in HL-60 cells (IC50 is about 200 ${\mu}g/ml$). The cytotoxic activity of N. Rhizoma in HL-60 cells was increased in a dose-dependent manner. We used several measures of apoptosis to determine whether these processes were involved in N. Rhizoma-induced apoptotic cell death. The high-dose (200 ${\mu}g/ml$) treatment of N. Rhizoma to HL-60 cells showed cell shrinkage, cell membrane blobbing, apoptotic bodies, and the fragmentation of DNA, suggesting that these cells underwent apoptosis. Treatment of HL-60 cells with N. Rhizoma time-dependently induced activation of caspase-3, caspase-8, and caspase-9 and proteolytic cleavage of poly(ADP-ribose) polymerase. Also, we investigated the effect of N. Rhizoma on cellular differentiation and proliferation in HL-60 cells. Differentiation and proliferation of HL-60 cells was determined through expression of CD11b and CD14 surface antigens using flow cytometry and nitroblue tetrazolium (NBT) assay, and through analysis of cell cycle using propidium iodide assay, respectively. N. Rhizoma induced the differentiation of HL-60 at the low-dose (100 ${\mu}g/ml$) treatment, as shown by increased expression of differentiation surface antigen CD11b, but not CDl4 and increased reducing activity of NBT. When HL-60 cells were treated with N. Rhizoma at concentration of $50{\mu}g/ml\;and\;100{\mu}g/ml$, NBT-reducing activities induced approximately 1.5-fold and 20.0-fold as compared with the control. In contrast, HL-60 cells treated with the N. Rhizoma-ATRA combination showed markedly elevated levels of 26.3-fold at $50{\mu}g/ml$ N. Rhizoma-0.1 ${\mu}M$ ATRA combination and 27.5-fold at 50 ${\mu}g/ml$ N. Rhizoma-0.2 ${\mu}M$ ATRA combination than when treated with N. Rhizoma alone or ATRA alone. It may be that N. Rhizoma plays important roles in synergy with ATRA during differentiation of HL-60 cells. DNA flow-cytometry indicated that N. Rhizoma markedly induced a G1 phase arrest of HL-60 cells. N. Rhizoma-treated HL-60 cells increased the cell population in G1 phase from 32.71% to 42.26%, whereas cell population in G2/M and S phases decreased from 23.61% to 10.33% and from 37.78% to 33.98%, respectively. We examined the change in the $p21^{WAF1/Cip1}\;and\;p27^{Kip1}$ proteins, which are the CKIs related with the G1 phase arrest. The expression of the CDK inhibitor $p27^{Kip1},\;but\;not\;p21^{WAF1/Cip1}$ were markedly increased by N. Rhizoma. Taken together, these results demonstrated that N. Rhizoma induces apoptotic cell death through activation of caspase-3, and potently inhibits the proliferation of HL-60 cells via the G1 phase cell cycle arrest in association with $p27^{Kip1}$ and granulocytic differentiation induction .

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.2
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• pp.481-489
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• 2005

Lonicerae Flos has antibacterial effects against Staphylococcus aureus, streptococci, pneumococci, Bacillus dysenterii, Salmonella typhi, and paratyphoid. It is an antiviral agent. The herb has a cytoprotective effect against $CCl_{4}-induced$ hepatic injury. It has antilipemic action, interfering with lipid absorption from the gut. Nowadays this herb is used mainly in the treatment of upper respiratory infections, such as tonsillitis and acute laryngitis. It is also used in the treatment of skin suppurations, such as carbuncles, and to treat viral conjunctivitis, influenza, pneumonia, and mastitis. Lonicerae Flos is dried flower buds of Lonicera japonica, L. hypoglauca, L. confusa, or L. dasystyla. But, for the most part, we use whole plant of Lonicera japonica, as a flower bud of it. And, little is known of the original copy of effects of whole plant, except for the 'Bon-Cho-Gang-Mok', which is written the effects of flower of Lonicera japonica are equal to effects of leaves and branch of it. The present study was conducted to evaluate the effect of flower and whole plant of Lonicera japonica on the regulatory mechanism of cytokines, inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX-2) for the immunological activities in Raw 264.7 cells. In Raw 264.7 cells stimulated with lipopolysaccharide (LPS) to mimic inflammation, flower and whole plant of Lonicera japonica water extracts inhibited nitric oxide production in a dose-dependent manner and abrogated iNOS and COX-2. Flower and whole plant of Lonicera japonica water extract did not affect on cell viability. To investigate the mechanism by which flower and whole plant of Lonicera japonica water extract inhibits iNOS and COX-2 gene expression, we examined the on phosphorylation of inhibitor ${\kappa}B{\alpha}$ and assessed production of $TNF-{\alpha}$, $interleukin-1{\beta}$ $(IL-1{\beta})$ and interleukin-6 (IL-6). Results provided evidence that flower and whole plant of Lonicera japonica inhibited the production of $IL-1{\beta}$, IL-6 and activated the phosphorylation of inhibitor ${\kappa}B{\alpha}$ in Raw 264.7 cells activated with LPS. These findings suggest that flower and whole plant of Lonicera japonica can produce anti-inflammatory effect, which may play a role in adjunctive therapy in Gram-negative bacterial infections, respectively.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.4
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• pp.766-783
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• 2006

This study describes philosophical background regarding '虛' in oriental medicine in an effort to understand the relationships among some of the Eastern philosophy in accordance with differentiated meanings in several resources by analyzing examples of '虛' in Huang Di Nei Jing. The various usages of '虛' used in Huang Di Nei Jing are as follows: naming; condition of pulse; emptiness; '太虛' which was referred universal space by Chinese ; insufficiency, lack or scarcity, deficiency ; and the description of vitality, mental faculties. 外丹修煉(training by external substances) had the attitude do that withdraw the death by taking external materials. The meaning of '虛'in 外丹修煉 is similar to that in oriental medicine in terms of deficiency. That is, both 外丹修煉 and the oriental medicine consider that the death and disease are caused by the deficiency of something. However, there also exists difference between 外丹修煉 and the oriental medicine. 外丹修煉 supplements through withdrawal prohibition due to the characteristic of unchangeability and stern or immortal while oriental medicine provides concrete object of deficiency. 精(essence of life), 氣(ki, functional activity), and fe(vitality) not only have been considered as basic component of human body, but they also have been an important subject of health preservation for longevity with health in Taoism and oriental medicine. In oriental medicine, 精 and 氣 have been perceived as physical basis of human body and 神 as controller. 內丹修煉(Training of internal active substances) 掠nds to return to '虛', the early state of life through individual training, and attempts to withdraw death through continuous recurrence. The oriental medicine and 內丹修煉 held great value of 神 among health preservation of 精, 氣, and 神. They seek theoretical basis from philosophical Taoism. However, '虛' in Taoism is different from that in training by internal substances and oriental medicine: '虛' in philosophical Taoism has metaphysical concept which refers overcome of life and death, but '虛' in 內丹修煉 and oriental medicine have empirical concept. '太虛' is considered as formless space where it is emp Dut filled with 氣. It is conceptualized with the premise of the relevant adaptation of human body to natural environment theory referring that the interaction between the heaven and the earth makes changes; all creation is originated , and human is affected by the interaction of the heaven and the earth. Furthermore, in $\ulcorner$運氣七篇$\lrcorner$ (Seven chapters described about the five circuit phases and the six atmospheric influences), the expression that the earth is in the center of '太虛' and huge amount of 氣 supports it proves that $\ulcorner$運氣七篇$\lrcorner$ adapts '渾天設'(Chaotic universe thee). In Taoism, '虛' is the grounds where all creation is generated in the optimal condition of Tao. As regards the aspect of mentality, it is the condition in which one can free from the dualistic concepts such as right and wrong, beauty and ugliness, life and death, and so on. Although the ultimate goal of oriental medicine, the achievement of longevity without sickness, might contrast with the Taoist belief that perceives life and death as the natural phenomena or the flowing of the 氣, and eliminates all international, the idea of Taoism that one should live substantial life with naivety, and make Harmony with the nature might be influential to the oriental medicine.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.4
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• pp.844-852
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• 2006

This study was to evaluate the effect of Yeongyupaedog-san (YGPDS) on mouse Thl and Th2 cells' differentiation and ovalbumin (OVA)-induced allergic inflammation. The proliferation of mouse CD4 T cells and the secretion of Th1/Th2 cytokines under the influence of YGPDS extract were measured as well as the amount of ${\beta}-hexosaminidase$ in RBL-2H3 cells and the levels of $TNF-{\alpha}$ and 1L-6 secretion in Raw264.7 cells. BALB/c mice were orally administered with YGPDS extract and simultaneously inoculated with OVA to induce allergic reaction and measure the level of total IgE, OVA-specific IgE and the production of IFN- g, IL-4, IL-5 by the spleen cells. When mouse CD4 T cell were stimulated with anti-CD3 and anti-CD28 for 48 hours in various concentrations of YGPDS extract, it increased proliferation of CD4 cells by 11% in $100\;{\mu}g/^{ml}$ concentration but it showed an inhibition by 37% at $200\;{\mu}g/^{ml}$ CD4 T cells under Th1/Th2 polarizing conditions for 3 days with YGPDS resulted in mild decrease of IFN- g in Thl cells and significant decrease of IL-4 in Th2 cells at $500\;{\mu}g/^{ml}\;and\;100\;{\mu}g/^{ml}$ by 18% and 21%, respectively. YGPDS extract had a dose-dependent inhibitory effect on antigen-induced release of ${\beta}-hexosaminidase$ in RBL-2H3 cells. Treatment of YGPDS extract on LPS stimulated Raw 264.7 cells showed dose-dependent decrease in TNF-n production. Oral administration of YGPDS extract on OVA-induced allergic mice showed an inhibitory effect on the levels of total serum IgE and OVA-specific IgE by 25% and 34% , respectively. Culture of spleen cells with OVA resulted in significant increase of IFN- g by 44% and significant decrease of IL-4 and IL-5 by 56%, and 24%, respectively. The results show that YGPDS does not strongly induce mouse T cells to transform into Thl or Th2 but it has an anti-allergic effect in vitro, and that it also corrects the unbalance between the reactions of Th cells in allergic diseases.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.6
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• pp.1467-1476
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• 2006

This study was to evaluate the effect of Bulhwangeumjeonggi-san (BS) on mouse Th1 and Th2 cells' differentiation and ovalbumin (OVA)-induced allergic inflammation. The proliferation of mouse CD4 T cells and the secretion of Th1/Th2 cytokines under the influence of BS extract were measured as well as the amount ${\beta}$-hexosaminidase in RBL-wH3 cells and the levels of TNF-${\alpha}$ and IL-6 secretion in Raw264.7 cells. BALB/c mice were orally administered with BS extract and simultaneously inoculated with OVA to induce allergic reaction and measure the level of total lgE, OVA-specific lgE and the production of IFN- g, IL-4, IL-5 by the spleen cells. When mouse CD4- T cell were stimulated with anti-CD3 and anti-CD28 for 48 hours in various concentrations of BS extract, it increased proliferation of CD4 cells by 14% in 50 ${\mu}g/m{\ell}$ concentration but it showed an inhibition in higher concentrations. CD4 T cells under Th1/Th2 polarizing conditions for 3 days with BS resulted in mild decrease of IFN- g in Th1 cells and mild increase of IL-4 in Th2 cell at 50 ${\mu}g/m{\ell}$ but the level of IL-4 decreased by 18% at 100 ${\mu}g/m{\ell}$. BS extract had a dose-dependent inhibitory effect on antigen-induced release of ${\beta}$-hexosaminidase in RBL-2H3 cells. Treatment of BS extract on LPS stimulated Raw 264.7 cells showed dose-dependent decrease in TNF-${\alpha}$ production. Oral administration of BS extract on OVA-induced allergic mice showed an inhibitory effect on the levels of total serum lgE and OVA-specific lgE by 50% and 55%, respectively. Culture of spleen cells with OVA resulted in significant increase of IFN- g by 25% and significant decrease of IL-4 and IL-5 by 53%, and 38%, respectively. The results show that BS does not strongly induce mouse T cells to transform into Th1 or Th2 but it has an anti-allergic effect in vitro, and that it also corrects the unbalance between the reactions of Th cells in allergic diseases.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.3
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• pp.591-595
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• 2007

There are eight kinds of formulas for invigorating the liver to cure its asthenic syndrome based on the types of preparation formulas Began-tang, Boganhwan, Bogansan, Bogandan, Boganeum, Boganjoo, Boganjeon and Bogango. Another formula called Bocheonghwan is to invigorate the green colour of liver. There are 22 kinds of Bogan-tang, 12 kinds of Boganhwan, 25 kinds of Bogansan, one kind of Bogandan, one kind of Boganeum, one kind of Boganjoo, one kind of Boganjeon, one kind of Bogango, and two kinds of Bocheonghwan. Combination of herbal medicines, carried out in formulas for invigorating the liver, consists of various kinds depending on medical scientists' personal experience in medical treatment without any general principles, which makes it difficult to apply it to clinical use. The objectives of this study lie in theoretical establishment of Bogan-tang for curing the asthenic syndrome of liver through analyzing the component medicines and combination principles of Hyeongok's Bogan-tang, and furthermore, maximizing the clinical use of Began-tang. This study analyzed the component medicines and combination principles of Hyeongok's Bogan-tang based on the theory for properties and tastes of herbal medicines from the ${\ulcorner}$yellow Emperor's Canon of Internal Medicine${\lrcorner}$ , the theory for principal herbal medicine, assistant herbal medicine, adjuvant herbal medicine, dispatcher herbal medicine, and the five elements doctrine. Hyeongok's Began-tang is an odd prescription, composed of 7 kinds of ingredients No.1 Radix Angelicas Sinensis (2don;7.5g), No.2 Rhizoma Chuanxiong (1don;3.75g), No.3 Radii Polygoni Multiflori (1don), No.4 Fructus Lycii (1don), No.5 Cortex Cinnamomi (1don), No.6 Rhizoma Gastrodiae (1don), and No.7 Radix Glycyrrhizae (1don). There are three methods for curing the asthenic syndrome of liver according to the five elements doctrine invigorating the liver, invigorating the kidney and purging the lung. First, if you suffer from the asthenic syndrome of the liver, you need to invigorate your liver. There are two available methods, including taste and property invigoration according to the theory for properties and tastes of herbal medicines. They each imply the pungent taste and the warm property invigorate the liver. In the case of taste invigoration, two herbal medicines with pungent taste, Radix Angelicas Sinensis and Rhizoma Chuanxiong, are combined into the principal and assistant herbal medicine, respectively. For property invigoration, two herDal medicines with the warm property, Cortex Cinnamomi and Rhizoma Gastrodiae, are combined into adjuvant herbal medicines. Secondly, if you suffer from the asthenic syndrome of the liver, you need to invigorate your kidney which is mother in the mother-child relationship in inter-promotion among the five elements. There are two methods to invigorate the kidney, including taste and property invigoration according to the theory for properties and tastes of herbal medicines. They each mean the bitter taste and the cold property invigorate the kidney. Therefore, it is important to use bitter herbal medicines for taste invigoration and cold ones for property invigoration. Both Differ and cold herbal medicines, Radix Polygoni Multiflori and Fructus Lycii, are combined into adjuvant herbal medicines. Lastly, if you suffer from the asthenic syndrome of the liver, you need to purge your lung which is an element being surpassed in the relationship between the elements surpassed and ones not surpassed in inter-restraint among the five elements. There are two methods to purge the lung, which include taste and property invigoration according to the theory for properties and tastes of herbal medicines. Taste invigoration means to purge the lung with pungent taste and property invigoration to purge the lung with warm property. Therefore, it is important to use pungent herbal medicines for taste invigoration and warm ones for property invigoration. Both pungent and warm herbal medicines, Radix Angelicas Sinensis and Rhizoma Chuanxiong, were combined to invigorate and purge the lung. In addition, Radix Glycyrrhizae are combined as dispatcher herbal medicine, harmonizing all the herbal medicines composing the formula. First, to cure the asthenic syndrome of the liver, the methods of invigorating the liver and the kidney, and purging the lung should be used according to the five elements doctrine. Secondly, herbal medicines appropriate for those treatment methods should be chosen according to the theory for properties and tastes of herDal medicine and thirdly, the combination of those herbal medicines should be carried out according to the theory for principal herbal medicine, assistant herbal medicine, adjuvant herbal medicine, dispatcher herbal medicine. As a good example, Hyeongok's Bogan-tangon is combined according to the above theories. In conclusion, this formula was created by applying to the theory for properties and tastes of herbal medicines.

• Journal of Preventive Medicine and Public Health
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• v.22 no.4 s.28
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• pp.578-590
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• 1989

This study was conducted to assess the equity in the regional insurance scheme through analysis of the computerized data from one regional insurance society and National Federation of Medical Insurance. We analysed the insurance contribution and benefit by the classes based on total and income-related contribution per household. The major findings of this study are as follows : 1. The average proportion of income-related contribution among the total was 39.2% and the upper classes show higher proportion of the income-related contribution. 2. The upper classes show higher health care utilization rate than the lower classes. It suggests that the lower classes have relatively large unmet medical needs. 3. The analysis through the Lorenz curve reveals that there exists transference of contributions from the upper to lower classes. But the cumulative percentage of insurance benefit is smaller than that of the number of the insured. It implies that regional medical insurance scheme in Korea has still some inequity in the context of social security principles.