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HISTOLOGICAL CHANGES IN THE ELONGATED BONE AFFECTED BY OSTEODISTRACTION OF THE MANDIBLE IN THE DOG (성견 하악골 절단 후 기계적 골 견인에 의해 형성된 골 신장부에 대한 시기별 조직학적 변화)

  • Baek, Sun-Ho;Ahn, Byoung-Keun;Park, Young-Ju;Park, Hee-Geon;Park, Jun-Woo;Rhee, Gun-Joo;Lee, Yong-Chan;Cho, Byoung-Ouck
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.27 no.5
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    • pp.404-416
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    • 2001
  • Purpose : Traditionally, the treatement of choice has been a bone grafting procedure to increase the length of bone in case of actual length discrepancy. But, bone grafting procedure has many disadvantages, for example, graft resorption, donor site morbidity, and so on. So, many trials have been performed to avert the use of autogenous bone graft via introducing new materials or methods. And, one of those trials has been realized by the development of a technique inducing bone lengthening by osteotomy (or corticotomy) and slow gradual distraction of the osteotomized segments. This new technique of bone lengthening dates back to the early 20th century. But, the majority of information concerning the biology of new bone formation during bone lengthening and technical details of the procedure were produced by extensive clinical and experimental studies performed by Ilizarov, a Russian surgeon. According to Ilizarov, with adequate blood supply, preservation of periosteum, rigid fixation of the osteotomized segments, and proper rate and rhythm of distraction, intramembranous bone rapidly develops within the distraction gap in the limb lengthening procedure. In the limb lengthening, many orthopedic surgeons try to observe the biologic and clinical principles recommended by Ilizarov. In the oral and maxillofacial region, however, not a few studies must be performed to apply this surgical technique in the clinical cases. Besides, the mechanism of bone formation in the distraction gap is not clear, yet. The purpose of this experiment was to scrutinize serially the histological changes in the elongated bone affected by osteodistraction of the mandibular body in an adult canine model. In addition, it was performed to confirm the presence of specific region(s) which was important in the bone formation in the gap through the observation of the expression pattern of osteocalcin and osteonectin with the immunohistochemical examination. Materials and Methods : The experimental and control specimens were obtained from seven adult male mongrel dogs weighing over 20kg. The distractors were custom-made linear extraoral devices and bicortical fixation screws were 2.3mm in diameter, 50mm in total length, 15mm in screw length. The distractors were devised to produce a linear gap of 0.75mm between two bony segments every $360^{\circ}$ turn of the rotation rod of the device. The mandibular body of the right side of each animal was corticotomized perpendicular to the occlusal plane and then two bony segments were separated completely by careful manipulation of the segments with bone forceps. The left side of each animal was left intact. This side was served as control. At sixth day after osteotomy and fixation of the segments were performed, distraction of the segments was commenced with a rate of 1.1mm/day and a rhythm of two/day for ensuing 7 days. The animals were euthanized at the 16th. 29th, and 44th day after the osteotomy. The bony specimens were decalcified, embedded in paraffin, sectioned $5{\mu}m$ thick and stained with H&E. The prepared specimens were examined under the light microscope. And, immunohistochemical examinations using anti-osteocalcin antibody (OC1, Biodesign, USA) and anti-osteonectin antibody (Haematologic Technologies Inc., Essex, VT) to locate the expressions of osteocalcin and osteonectin, respectively, were performed. Results : 1. New bone was observed already at the 16th. day after osteotomy. This suggests that new bone formation in osteodistraction was commenced at an early stage of the regenerative process. But, radiologically and microscopically, bony union was not completed in the distraction gap at the 44th. day after osteotomy. Therefore, rigid fixation must be maintained between the bony fragments till the complete bony union is confirmed clinically rather than one month or so after the completion of distraction.

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Effect of various abutment systems on the removal torque and the abutment settling in the conical connection implant systems (원추형 연결 임플란트에서 지대주 종류에 따른 나사풀림과 침하현상에 관한 연구)

  • Lee, Jin-Seon;Lee, Joon-Seok
    • The Journal of Korean Academy of Prosthodontics
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    • v.50 no.2
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    • pp.92-98
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    • 2012
  • Purpose: The aim of this study was to evaluate the effects of different abutment materials on abutment screw loosening and settling-down effect in conical connection type implant system. Materials and methods: Three types of abutment, cementation, gold UCLA, and metal UCLA abutment were used. Two UCLA groups were fabricated in a similar pattern to cementation abutment. Type III gold alloy and Nickel-Chromium alloy was used for casting gold UCLA abutment and metal UCLA abutment, respectively. Fixture and abutment were tightened to 30 Ncm by using digital torque controller and re-tightening was conducted with same force after 10 minutes. Digital torque gauge was used to measure loosening torque and fixture/abutment length was measured by digital micrometer. Dynamic loads between 25 N and 250 N were applied with $0^{\circ}$ angle to the abutment axis. After loading, fixture/abutment length was re-measured and amount of settlement was calculated. Loosening torque value was also measured for comparison Results: All three groups showed significant differences of length when comparing before and after loading, but there was no significant difference of settling amount in all groups. Loosening torque values were significantly decreased when comparing before and after loading in all groups($P$<.05). However, there was no significant difference in loss of loosening torque values when compared to groups. Conclusion: In internal conical connection type implants, dynamic load affected on settlement and loosening torque of implant, but there was no differences between abutments materials. Likewise gold UCLA abutment, metal UCLA abutment might be able to withstand functional load.

Two Crystal Structures of Fully Dehydrated $Ag_{12-2x}Co_x-A (x = 3 and 4.5)$ (완전히 탈수한 $Ag_{12-2x}Co_x-A$ (x = 3 및 4.5)의 결정구조)

  • Seung Hwan Song;Duk Soo Kim;Jong Yul Park;Un Sik Kim;Yang Kim
    • Journal of the Korean Chemical Society
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    • v.32 no.6
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    • pp.520-527
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    • 1988
  • The crystal structures of $Co^{2+}\;and\;Ag^+\;exchanged\;zeolite\; A,\; Ag_6Co_3$-A(a = 12.131(5)$\AA$) and $Ag_3Co_{4.5}$-A(a = 12.145(1)$\AA$), have been determined by single crystal X-ray diffraction techniques. Both structures were solved and refined in the cubic space group Pm3m at 21(1)$^{\circ}C$. Full-matrix leastsquares refinement converged to the final error indices of R1 = 0.045 and R2 = 0.041 for $Ag_3Co_{4.5}-A,\; and\; R1 = 0.066\; and\; R2 = 0.076\; for\; Ag_6Co_3$-A using the 258 and 189 reflections, respectively, for which I > 3$\sigma$(I). Both structures indicate that CO(Ⅱ)ions are coordinated by three framework oxygens; the Co(II) to O(3) distances are 2.118(4)$\AA$ for $Ag_3Co_{4.5}$-A and 2.106(1)$\AA$ for $Ag_6Co_3-A$, respectively. In each structure, the angle substended at Co(II), O(3)-Co(II)-O(3) is ca 120°, close to the idealized trigonalplanar value. $Co^{2+}$ ions prefer to 6-ring sites and $Ag^+$ ions prefer to 8-ring site when total number of cations is more than 8. The crystals of hydrated and dehydrated $Ag_{12-2x}Co_x-A (x > 4.5)$ had no crystalline diffraction pattern, indicating the apparent exchange limit of $Co^{2+}\; into\; Ag_{12}-A\; is\; 4.5 Co^{2+}$ ions per unit cell. $Co^{2+}$ ions hydrolyze $H_2O$ molecules and $H_3O^+$ concentraction is accumulating. These $H_3O^+$ ions destroy the zeolite structures.

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Different Metabolic Patterns of Parkinsonism: Analysed by Statistical Parametric Mapping (통계적 파라미터를 이용한 Parkinsonism의 Metabolic pattern 분석)

  • 주라형;김재승;최보영;문대혁;서태석
    • Progress in Medical Physics
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    • v.14 no.2
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    • pp.108-123
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    • 2003
  • The purpose of this study is to evaluate the contribution of $^{18}$ F-FDG brain PET in the differentiating Idiopathic parkinson's diesease (IPD), progressive supranuclear palsy (PSP), and multiple system atrophy (MSA). We studied 24 patients with parkinsonism : 8 patients (mean age 67.9$\pm$10.7 y: M/F : 3/5) with IPD, 9 patients (57.9$\pm$9.2 y : M/F : 4/5) with MSA and 7 patients (67.6$\pm$4.8 y : M/F 3/4) with PSP. All patients with parkinsonism and 22 age-matched normal controls underwent $^{18}$ F FDG PET in 3D mode after the injection of 370 MBq $^{118}$ F FDG. The patients with IPD, MSh and PSP were compared with a normal control group by a two-sided t-test of SPM99 (uncorrected P<0.001, extent threshold>100 voxel). All three parkinsonism groups, showed significant hypometabolism in the cerebral neocortex compared to the normal control group. However, the three groups displayed different metabolism in the subcortical structure, brain stem, and cerebellum. In IPD, there was no significant hypometabolism in the putamen, brain stem and cerebellum. However, MSA patients showed significant hypometabolism in the striatum, pons, and cerebellum compared to the normal controls and IPD patients. In addition, PSP showed significant hypometabolism in the caudate nuclei, the thalamus, midbrain, and the cingulate gyrus compared to the normal controls, the IPD, and MSA groups (IPD vs Normal sensitivity/specificity : 75%/l00%, MSA vs Normal sensitivity/specificity :100%/87%, PSP vs Normal sensitivity/specificity : 86%/94%). Our results show that the regional metabolism of IPD, MSA, and PSP is different mainly in the striatum, thalamus, brain stem and cerebellum. An assessment of the $^{18}$ F-FDG PET scan images using SPM may be a useful adjunct to a clinical examination in making a differential diagnosis of Parkinsonism.

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노쇠중인 밀 잎에서 scorbate-Glutathione회로 관계 효소의 발달에 대한 Benzyladenine의 효과

  • 장창덕
    • Journal of Plant Biology
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    • v.38 no.1
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    • pp.47-54
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    • 1995
  • The present study performed the isolation of cytosolic ascorbate peroxidase (APX) isozymes and analyzed the pattern of their activity development and also investigated the change in some other enzyme activities related to the ascorbate-glutathione pathway from the senescing wheat leaves. The aim of this work is to examine the possibility that in the cytoplasm of wheat leaves the ascorbate-glutathione pathway p!ays a significant role in relation to leaf senescence involving an $H_2O_2$ accumulation and then to show the effect of benzyladenine (BA) on that pathway. During the leaf senescence characterized by increases in ChI breakdown and H202 accumulation under the 4-day dark incubation of matured leaf segments; i) no significant increase of total cytosolic APX was observed, ii) a dehydroascorbate reductase (DHAR) activity was decreased rapidly, iii) a slight increase of glutathione reductase (GR) activity occurred. In the BA-treated leaves; however, i) the total activity of APX increased conspicuously, ii) the decrease of DHAR activity was relatively inhibited, iii) the GR activity increase was more enhanced, and iv) the decrease of ascorbate content and the increase of H202 content were retarded as compared with those of control leaves. Three isozymes of cytosolic APX were found by using a native-electrophoretic gel in senescing wheat leaves and two of them occurred with major activity. In the developmental patterns of cytosolic APX isozymes, only two isozyme bands ("a" and "b") appeared with almost constant activity through 4 days of incubation in the control leaves, while one additional weak isozyme band ("c") and a little increase of "b" isozyme activity were detected in the BA-treated leaves. EspeciaUy, the development of "a" isozyme activity increased remarkably compared with that of control leaves. The increased capacity for peroxide scavenging due to the enhanced activity of all 3 enzymes (APX, DHAR, GR) participating in the ascorbate-glutathione pathway in BA-treated leaves suggested that this pathway might playa significant role in the processes related to the wheat leaf senescence.scence.

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Quality characteristics of Yanggaeng with Momordica charantia powder (여주 분말을 첨가한 양갱의 품질 특성)

  • Lee, Seon-Ho;Hong, Eun-Jin;Cho, Young-Je
    • Food Science and Preservation
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    • v.22 no.3
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    • pp.335-344
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    • 2015
  • The objective of this study was to determine the rheological characteristics and sensory evaluation of yanggaeng by using the functional properties of Momordica charantia. Dried Momordica charantia was roasted at $180^{\circ}C$ to remove its bitter taste. The results of the study were as follows : The phenolic compound of Momordica charantia was $6.30{\pm}0.17mg/g$ in water extracts, which was the highest extractive rate. The anti-oxidant activity of non-roasted and roasted samples was determined in various phenolic concentrations at $50{\sim}200{\mu}g/mL$. The DPPH activities of non-roasted and roasted Momordica charantia water and ethanol extracts were 74.06~92.71% and 86.06~94.07%, respectively. The ABTS were 36.26~98.03% and 67.02~99.60% in water and ethanol extracts, respectively. The anti-oxidant protection factor of water and ethanol extracts were 2.19~2.25 PF and 2.20~2.36 PF respectively, and TBARS were 13.81~40.97% and 23.32~82.47%, respectively. The anti-oxidant activity of ethanol extracts was higher than that of water extracts at low phenolics concentration of $50{\mu}g/mL$, while the roasted sample was higher than the sample that was not roasted. The ${\alpha}$-glucosidase inhibition activity of non-roasted Momordica charantia ethanol extracts was higher than that of water extracts, showing that there were increasing pattern depending on the increases in the phenolics concentration of Momordica charantia. The texture, such as hardness, springiness, cohesiveness, chewiness and color changed in proportion to the concentration of Momordica charantia powder. Moreover sensory characteristics, such as color, flavor, taste, texture and overall acceptability of the non-roasted sample changed in proportion to the concentration of Momordica charantia powder. However, in the roasted sample, the sensory characteristics was improved at a lower concentration (below 1%). Thus, when the yanggang was prepared by Momordica charantia powder as a minor ingredient, it would be desirable to add it after roasting, in consideration of its functional and sensory properties, and at appropriate concentration on below 1%.

Quality characteristics and biological activity of Yanggaeng with Glechoma hederacea var. longituba Nakai powder (긴병꽃풀의 생리활성과 이를 첨가한 양갱의 품질특성)

  • Lim, Su-Bin;Park, Ki-Tae;Lee, Eun-Ho;Kim, Byung-Oh;Lee, Seon-Ho;Kang, In-Kyu;Cho, Young-Je
    • Food Science and Preservation
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    • v.24 no.2
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    • pp.206-214
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    • 2017
  • The objective of this study is to investigate the quality characteristics of Yanggaeng by using the functional properties of Glechoma hederacea (GH). Sample was dried at $50^{\circ}C$ dry oven. The results of the study were as follows : The Phenolic compounds of GH was $12.99{\pm}0.3mg/g$ in water extract (GHWE), $3.14{\pm}0.07mg/g$ in 70% ethanol extract (GHEE). The antioxidant activity of GH was determined in various phenolic concentrations at $50-200{\mu}g/mL$. DPPH activities of GHWE and GHEE were 77.16-78.24% and 73.04-77.00%, respectively. The ABTS were 84.35-99.75% and 83.74-99.55%. The anti-oxidant protection factor (PF) were 1.54-1.62 PF and 1.62-2.09 PF and TBARS were 42.93-94.09% and 91.05-95.19%, respectively. Tyrosinase inhibitory activity of GHEE increased concentration dependently. Hyaluronidase inhibition activity of GHEE and GHWE, showing that there were increasing pattern depending on the increases in the phenolics concentration of GH. In texture, Hardness and springiness were significantly different in the control and 2% groups, but cohesiveness and chewiness did not show any significant difference. In color, L value decreased in proportion to concentration, and a and b values did not change. Sensory characteristics showed that the 1% group had the highest score and the 2% group had the lowest score. Thus, when the GHP Yanggaeng was prepared, in consideration of its sensory characteristics, and at appropriate concentration on below 1%.

A Study on Dementia Alzheimer's type published to chinese magazine (중의잡지(中醫雜誌)에 보고(報告)된 Alzheimer형(型) 치매에 대(對)한 고찰(考察))

  • Chae, Jong-gul;Lee, Sang-ryong
    • Journal of Haehwa Medicine
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    • v.10 no.1
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    • pp.453-469
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    • 2001
  • This study attempted to analyze the contents of the research papers concerning the diagnosis and treatment of Alzheimer-type dementia presented in the magazine of Chinese Medicine published in China over the period between 1998 and 2000. As a result, the following conclusion was drawn: 1. The Chinese medical category of Alzheimer-type dementia includes amnesia, dementia, stupidity, depression symptom complex, insanity and the like and uses the as the criterion for diagnosis and treatment effect evaluation. 2. The clinical symptoms of Alzheimer-type dementia include lowered intelligence, deterioration of memory, understanding and judgemental power, retardation of the reaction, emotional changes, character changes, behavioral changes and the like and are divided into mild, medium and serious according to the degree of symptom. 3. From the perspective of Bon-Heo-Pyo-Shil(state of deficient vital essence and excessive pestilential vapor), the pattern of Byun-Sung(identification) is divided into deficiency symptom complex, excessive symptom complex and indiscernible fullness and emptiness. The deficiency symptom complex includes deficiency of the liver and kidney, deficiency of essence of the kidney, deficiency of the reservoir of marrow and the like. The excessive symptom complex includes internally blocked stagnant blood, blocking of the passageway due to turbid phlegm, blood stasis due to stagnation of chi, and the like. The indiscernible fullness and emptiness symptom complex includes the deficiency of essence of kidney, blocking of the passageway due to stagnant phlegm, blood stasis due to the deficiency of kidney, blood stasis due to the deficiency of heart and the like. 4. The therapeutics and' prescription of Alzheimer-type dementia include the following: Bo-Shin-Ik-Su-Tang for tonifying the kidney, replenishing the marrow and plugging the essence; Ki-Guk-Ji-Hwang-Hwan-Ga-mi for reinforcing the vital essence of the liver and kidney; Kwi-Bi-Tang-Hap-Yang-Shim-Tang for invigorating the functioning of the spleen and nourishing the heart; Hyel-Bu-Chuk-Eo-Tang-Ga-Mi for activating the blood and resolving the stagnancy of the blood; Bo-Yang-Hwan-Oh-Tang for replenishing chi, activating the blood and resolving the stagnancy of the blood; Beoh-Kwang-Mong-Sung-Tang for invigorating the functioning of the spleen, replenishing the kidney, resolving the phlegm and enlivening the brain; n-Dam-Tang-Ga-Mi for invigorating the functioning of the spleen, replenishing chi, and removing the phlegm and unclogging the passageway); Se-Shim-Tang-Ga-Mi for removing the stagnancy of the liver and resolving the phlegm; and the like. 5. The research papers on, the medication cases of Alzheimer-type dementia understand the pathology of Alzheimer-type dementia from a consistent perspective. They view the pathology of Alzheimer-type dementia as the disease of Bon-Heo-Pyo-Shil(state of deficient vital essence and excessive pestilential vapor) with the combination of the deficiency of essence of the kidney, the deficiency of the brain marrow, blood stasis and blocked phlegm and the like and recommend the prescription of using Bo and Sa simultaneously for treating Alzheimer-type dementia. 6. The research papers on the medication cases of Alzheimer-type dementia reported that the use of creative prescriptions such as Si-Sam-Hang-Ji-Tang, Ji-Yung-Tang, Ka-Mi-Yunh-Ji-Hwan, Ja-Sin-Hwal-Hyel-Tang, Kal-Chang-Ik-Ji-Tang, Ho-Su-Bok-Ji-Tang, Kun-Noe-Ok-Ji-Hap-Je and the like led to the average high efficacy of 85.5%.

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A Study on the Characteristics of Descriptions of the Perspiration in "Hwangjenaegyeong(黃帝內經)" (황제내경(黃帝內經)에 보이는 한(汗)관련 서술(敍述)의 특징(特徵)에 대한 고찰(考察))

  • Lyu, Jeong-Ah;Jang, Woo-Chang;Baik, You-Sang;Jeong, Chang-Hyun
    • Journal of Korean Medical classics
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    • v.23 no.2
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    • pp.205-223
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    • 2010
  • In Korean Traditional Medicine(abbreviated to K.T.M.), hyperhidrosis and anhidrosis are the targets of the medical treatment. Furthermore sweating appearance is also one of the important symptoms which explain a particular situation of the patient in K.T.M. And at "Sanghanron(傷寒論)" which is a traditional chief clinical bible written by Jang Gi(張機) later Han dynasty(漢代) in China made full use of the various kinds of diaphoresis[汗法] as a main medical treatment with purgation therapy[下法] and emetic therapy[吐法]. So the sweat in itself not only is the disease, but also is one of the symptoms explain a disease pattern. This thesis inquires into "Hwangjenaegyeong(黃帝內經)" referring to sweat which is the origin of recognition to the sweat in K.T.M. Some theses similar to this research had been made progresses and already reported, but most of them have classified the contents into biology, pathology, diagnosis, treatment after the model of western medical theory. In the aspect of comparative studying with other literature and clinic practical using, we found characteristics of referring to sweat in "Hwangjenaegyeong(黃帝內經)". And we classify the characteristics into some categories as follows. 1. There are some terms which make a title including sweat and symbolize the characteristics, for example sweat of soul[魄汗], sweat of death[絶汗], sweat of streaming[灌汗], sweat of weakness[白汗], sweat of sleep[寢汗], sweat of bright and heat[炅汗], sweat of kidney[腎汗], sweat of escaping[漉汗], cold sweat[寒汗], sweat on the head[頭汗], hyperhidrosis[多汗], heavy sweat[大汗]. But there aren't spontaneous sweat[自汗] or sweat like a thief[盜汗] which are the normal terms referring to sweat in history of K.T.M. And there are several descriptions about sweat appearance such as sweating in half of body[汗出偏沮], sweating in the rear end and thigh and knee[汗出尻陰股膝], hyperhidrosis in the neck and aversion to wind[頸多汗惡風], hyperhidrosis in the head and face and aversion to wind[頭面多汗惡風], cannot stopping the sweating under head[頭以下汗出不可止], make a person sweat to one's feet[令汗出至足], sweating like escaping[漯漯然汗出], sweating like soaking[汗出如浴], sweating become moist[汗出溱溱], hardly escaping sweat[汗大泄], escaping sweating[漉漉之汗], sweat moisten the pores [汗濡玄府], ceaseless sweating like pouring[汗注不休] sweating like pouring and vexation[汗注煩心], damp with sweat[汗汗然], sweating spontaneously[汗且自出], removal of fever with sweat drying[熱去汗稀]. That can be divided into sweat region and sweat form. 2. There are detailed explanations of the principle of perspirations caused by hot weather, hot food, hard working and meeting damp pathogen. 3. There are some explanations of the principle of removing fever due to the excessive heat from internal and external body through sweating by replenishing the body fluid. And many descriptions about overcoming the febrile disease by dropping temperature through sweating and many diaphoresis for curing. 4. There are some descriptions about five Jang organs perspirations and attachment of five mucous body fluid to five Jang organs. 5. There are pathogenic progresses after sweating affected by the Six Atmospheric Influences and water. And detailed explanations of disease mechanism a sweat leading to another disease. 6. There are descriptions about various sweat absent situations.

Amino Acid Biosynthesis and Gene Regulation in Seed (종자내 아미노산 합성 조절 유전자에 관한 연구)

  • ;;;;;Fumio Takaiwa
    • Proceedings of the Botanical Society of Korea Conference
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    • 1996.07a
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    • pp.61-74
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    • 1996
  • Human and monogastric animals can not synthesize 10 out of the 20 amino asids and therefor need to obtain these from their diet. The plant seed is a major source of dietary protein. It is particular important in their study to increase nutritional quality of the seed storage proteins. The low contents of lysine, asparagine and threonenein various cereal seeds and of cystein and methionine. In legume seeds is due to the low proportions of these amino acids in the major storage proteins, we have tried to apply the three strategies; (1) mutagenesis and selection of specific amino acid analogue resistance, (2) cloning and expression study of lysine biosynthesis related gene, (3) transfomation of lysine rich soybean glycinin gene. The 5-methyltryptophan (5MT) resistant cell lines, SAR1, SAR2 and SAR3 were selected from anther derived callus of rice (Oryza sativa L. "Sasanishiki"). Among these selected cell lines, two (SAR1 and SAR3) were able to grow stably at 200 mg/L of 5MT. Analysis of the freed amino acids in callus shows that 5MT resistant cells (SAR3) accumulated free tryptophan at least up to 50 times higher than those that of the higher than of SAS. These results indicated that the 5MT resistant cell lines are useful in studies of amino acid biosynthesis. Tr75, a rice (Oryza sativa L., var. Sasanishiki) mutant resistant to 5MT was segregated from the progenies of its initial mutant line, TR1. The 5MT resistant of TR75 was inherited in the M8 generations as a single dominant nuclear gene. The content of free amino acids in the TR75 homozygous seeds increased approximately 1.5 to 2.0 fold compared to wild-type seeds. Especially, the contents of tryptophan, phenylalanine and aspartic acid were 5.0, 5.3 and 2.7 times higher than those of wild-type seeds, respectively. The content of lysine is significantly low in rice. The lysine is synthesized by a complex pathway that is predominantly regulated by feedback inhibition of several enzymes including asparginase, aspatate kinase, dihydrodipicolinat synthase, etc. For understanding the regulation mechanism of lysine synthesis in rice, we try to clone the lysine biosynthetic metabolism related gene, DHPS and asparaginase, from rice. We have isolated a rice DHPS genomic clone which contains an ORF of 1044 nucleotides (347 amino acids, Mr. 38, 381 daltons), an intron of 587 nucleotides and 5'and 3'-flanking regions by screening of rice genomic DNA library. Deduced amino acid sequence of mature peptide domain of GDHPS clone is highly conserved in monocot and dicot plants whereas that of transit peptide domain is extremely different depending on plant specie. Southern blot analysis indicated that GDHPS is located two copy gene in rice genome. The transcripts of a rice GDHPS were expressed in leaves and roots but not detected in callus tissues. The transcription level of GDHPS is much higher in leaves indicating enormous chloroplast development than roots. Genomic DNA clones for asparaginase genes were screened from the rice genomic library by using plaque hybridization technique. Twelve different genomic clones were isolated from first and second screening, and 8 of 12 clones were analyzed by restriction patterns and identified by Southern Blotting, Restriction enzyme digestion patterns and Southern blot analysis of 8 clones show the different pattern for asparaginase gene. Genomic Southern blot analysis from rice were done. It is estimated that rice has at least 2-3 copy of asparaginase gene. One of 8 positive clones was subcloned into the pBluescript SK(+) vector, and was constructed the physical map. For transformation of lysine rich storage protein into tobacco, soybean glycinin genes are transformed into tobacco. To examine whether glycinin could be stably accumulated in endosperm tissue, the glycinin cDNA was transcriptionally fused to an endosperm-specific promotor of the rice storage protein glutelin gene and then introduced into tobacco genomic via Agrobacterium-mediated transformation. Consequently the glycinin gene was expressed in a seed-and developmentally-specific manner in transgenic tobacco seeds. Glycinin were targeted to vacuole-derived protein bodies in the endosperm tissue and highly accumulated in the matrix region of many transgenic plant (1-4% of total seed proteins). Synthesized glycinin was processed into mature form, and assembled into a hexamer in a similar manner as the glycinin in soybean seed. Modified glycinin, in which 4 contiguous methionine residues were inserted at the variable regions corresponding to the C - teminal regions of the acidic and basic polypeptides, were also found to be accumulated similarly as in the normal glycinin. There was no apparent difference in the expression level, processing and targeting to protein bodies, or accumulation level between normal and modified glycinin. glycinin.

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