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Mouse model system based on apoptosis induction to crypt cells after exposure to ionizing radiation (방사선에 전신 조사된 마우스 음와 세포의 아포토시스 유도를 이용한 생물학적 선량 측정 모델 개발 연구)

  • Kim, Tae-Hwan
    • Korean Journal of Veterinary Research
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    • v.41 no.4
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    • pp.571-578
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    • 2001
  • To evaluate if the apoptotic fragment assay could be used to estimate the dose prediction after radiation exposure, we examined apoptotic mouse crypt cells per 1,000 cells after whole body $^{60}Co$ $\gamma$-rays and 50MeV ($p{\rightarrow}Be^+$) cyclotron fast neutron irradiation in the range of 0.25 to 1 Gy, respectively. The incidence of apoptotic cell death rose steeply at very low doses up to 1 Gy, and radiation at all doses tigger rapid changes in crypt cells in stem cell region. These data suggest that apoptosis may play an important role in homeostasis of damaged radiosensitive target organ by removing damaged cells. The curve of dose-effect relationship for the data of apoptotic fragments was obtained by the linear-quadratic model $y=0.18+(9.728{\pm}0.887)D+(-4.727{\pm}1.033)D^2$ ($r^2=0.984$) after $\gamma$-rays irradiation, while $y=0.18+(5.125{\pm}0.601)D+(-2.652{\pm}0.7000)D^2$ ($r^2=0.970$) after neutrons in mice. The dose-response curves were linear-quadratic, and a significant dose-response relationship was found between the frequency of apoptotic cell and dose. These data show a trend towards increase of the numbers of apoptotic crypt cells with increasing dose. Both the time course and the radiation dose-response curve for high and low linear energy transfer (LET) radiation modalities were similar. The relative biological effectiveness (RBE) value for crypt cells was 2.072. In addition, there were significant peaks on apoptosis induction at 4 and 6h after irradiation, and the morpholoigcal findings of the irradiated groups were typical apoptotic fragments in crypt cells that were hardly observed in the control group. Thus, apoptosis in crypt cells could be a useful in vivo model for studying radio-protective drug sensitivity or screening test, microdosimetric indicator and radiation-induced target organ injury. Since the apoptotic fragment assay is simple, rapid and reproducible in the range of 0.25 to 1 Gy, it will also be a good tool for evaluating the dose response of radiation-induced organ damage in vivo and provide a potentially valuable biodosimetry for the early dose prediction after accidental exposure.

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Screening of Chemicals on Bacterial Brown Blotch Caused by Pseudomonas tolaasii on Flammulina velutipes (팽이버섯 세균성갈색무늬병(Pseudomonas tolaasii) 방제약제 선발)

  • Lee, Hyun-Uk;Kim, Tae-Sung;Park, Hyean-Cheal;Song, Keun-Woo;Shin, Won-Kyo;Moon, Byung-Ju
    • The Korean Journal of Mycology
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    • v.27 no.2 s.89
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    • pp.164-169
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    • 1999
  • This study was carried out in an attempt to select chemicals such as zeolite granules, chitosan, wood-vinegar, $Ca(OCl)_2$, NaOCl, against bacterial brown blotch caused by Pseudomonas tolaasii on Flammulina velutipes in laboratory and mushroom unit. Results obtained using these chemicals were summarized. Inhibitory effect on $Ca(OCl)_2$ and wood-vinegar with 0.5% concentration was shown to caual pathogen, P. tolaasii, by slight damage on the mycelial growth of F. velutipes. Those materials were recognized as promising one for control of bacterial brown bloth on F. velutipes. Disease incidence in control was the highest value as 43.8%; whereas chemical treatment was estimated as 14.6% with 0.5% of $Ca(OCl)_2$. Disease incidence was inhibited about 20% in chemical treatment with 0.5% of wood-vinegar, 1.0% of $Ca(OCl)_2$, and 26.1% with 1.0% of wood-vinegar. For the inhibition effects of chemical concentrations, it was effective at the low concentration which was 0.5% rather than that of 1.0%. Quality of mushrooms was significantly improved, and yields was also increased by 30% in the chemical treatment. In case of $Ca(OCl)_2$ treatment at 1.0% concentration, the yields was increased by 35.6% showed the highest value among tested chemicals. However, the overall effects including disease incidence, quality and yields suggests that $Ca(OCl)_2$ is relatively more effective than wood-vinegar, and the optimal concentration controlling the disease was $0.5{\sim}1.0%$ with $Ca(OCl)_2$ and 0.5% with wood-vinegar.

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Studies on Antimicrobial Susceptibility and Characteristics of R-plasmids and Antigens of High-level Gentamicin Resistant Enterococcus faecalis (Gentamicin 고도내성 Enterococcus faecalis균주의 항균제감수성, R-플라스미드 및 항원의 특성연구)

  • Kang, Hyun
    • Biomedical Science Letters
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    • v.1 no.1
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    • pp.55-72
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    • 1995
  • Forty gentamicin-resistant isolates of Enterococcus faecalis were selected from various clinical materials, determined their antimicrobial susceptibility, and studied there R-plasmid characteristics and polypeptide patterns. All of the isolates were susceptible to vancomycin. The MICs($\mu$/ml) of antimicrobial agents to the isolates were as follows; the MIC of gentamicin was 128 and $\geq$2040, ampicillin 1 and 1, chlorarmphenicol 2 and 8, erythromycin 32 and 256, and vancomycin 1 and 2. E. faecalis HL-1 strain had 8 plasmid DNA elements, HL-2 and HL-3 strains had 6, HL-4 had 7, HL-5 had 4, and HL-6 had 5. The 51.7 Kb of gentamicin resistance plasmid DNA was conjugally transferred from two strains of E. faecalis HL-1 and HL-6 to S. aureus SK 982. The plasmid transfer frequency between S. aureus SK 982 and E. faecalis HL-1 or E. faecalis HL-6 was 6.3$\times10^{-4} and 3.7$\times10^{-5}$, respectively. Plasmid curing ratio after the treatment of ethidium bromide(10$\mu$/ml) to E. faecalis tarnsconjugants R-1 and R-6 were about 51% and 67%, respectively. The tetracycline gene was located in 2.15 Kb plasmid of E. faecalis HL-1, but it was not found in the E. faecalis HL-6 by Southern blot analyses. The antigenic components of E. faecalis HL-1, HL-6, R-1 and R-6 strains were analyzed by SDS-PAGE and immunoblotting. The E. faecalis strains had 7 to 16 polypeptide bands, however their major proteins were 97.8 and 26.8 Kd. At the Immunoblotting, 97.8, 95.8, 74.8, 63.5, 33.7 and 26.8 Kd polypeptides of the strains showed major antigenic activities with patient's sera infected intra-abdominally with an E. faecalis strain.

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Comparative Study on Immunological Markers Between Human Immunodeficiency Virus(HIV)-Infected and Normal Persons in Korea (국내 Human Immunodeficiency Virus(HIV) 감염자와 정상인의 면역학적 표지인자 비교연구)

  • 최병선;박용근;류재천;신영오
    • Biomedical Science Letters
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    • v.1 no.1
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    • pp.27-35
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    • 1995
  • Several studies showed that the immunological factors such as CD4+ cell number, CD4%, CD8+ cell number and CD4/CD8 ratio and the serological factors such as, ${\beta}^2$-microglobulin(${\beta}^2$-MG), neopterin, soluble CD4, and soluble CD8 are related to the risk of development of AIDS. Especially, the CD4+ cell counts have been used to monitor progresson of HIV disease, to stratify, and to follow patients in clinical trials. Recently, the Centers for Disease Control and Prevention(CDCP) in USA has made the CD4+ cell count as a part of the classification of HIV disease. It is composed of 3 categories such as 1, 2, and 3 which asr $\geq$ 500/$mm^3$, 200/$mm^{3} $\geq$ and < 500/$mm^3$, and < 200/$mm^3$, respectively. In this study, to estimate the differences of immunological factors between HIV-infected and normal human groups in Korea, CD4+ T and CD8+ T cells, and the CD4/CD8 ratio were measured in 185 HIV-infected subjects and 140 healthy adult subjects. The lymphocyte subsets such as CD4+ T and CD8+ T were analysed by flow cytometer(FACStar) with two-color immunofluorescent stain using monoclonal antibodies such as anti-CD4 and anti-CD8 antibodies. The absolute numbers and percentages of CD4+ T and CD8+ T and the CD4/CD8 ratio of HIV infected persons were $462\pm{277}/mm^3$, $18.2\pm7.7%$, $1,170\pm{534}/mm^3$, $47.0\pm10.6%$ and $0,43\pm0.26 whereas those of uninfected persons were $886\pm{299}mm^3$, $32.9\pm{7.0%}, 730{\pm}259/mm^3$, $26.8\pm6.4%$ and $1.31\pm0.46$(P<0.01). In addition, estimating the reference values of peripheral blood lymphocyte subsets of Korean, the absolute numbers and percentages of CD4+ T and CD8+ T and the CD4/CD8 ratio of 140 healthy adults persons were measured and compared with those of foreigners. The reference ranges of CD4+ T cells, CD8+ T cells, CD4%, CD8%, and the CD4/CD8 ratio and 1.31$\pm$0.46, respectively. The significant differences were not observed when compared with those of foreigners. However a little difference was observed in the percentages of CD4+ T and the absolute numbers of CD8+ T between the normal values of Korean and those of foreigners were $43.6\pm8.9%$, $560\pm{230}/mm^3$. This result can also be useful as a basic data for the treatment and surveillance of HIV-infected patients in Korea.

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Development of the Bitterling, Acanthorhodus asmussi (Cyprinidae) with Note on Minute Tubercles on the Skin Surface (큰납지리의 난발생(卵發生)과 자어(仔魚)의 발육(發育) 및 자어(仔魚)의 표피상돌기(表皮上突起))

  • Suzuki, Nobuhiro;Jeon, Sang-Rin
    • Korean Journal of Ichthyology
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    • v.1 no.1_2
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    • pp.73-82
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    • 1989
  • The development of eggs and larvae, and minute scale-like tubercles on the skin surface of the larval Acanthorthodeus asmussi from Korea were observed in the laboratory. The egg was nearly ovoid-shaped. The number of egg averaged 195 per an oviposition. The morphological character of larval development was relatively similar to those of Acheilognathus tabira tabira, A. tabira subsp. (a), A. yamatsutae, A. moriokae, A. cyanostigma and A. rhombeus. The larvae of this species had many scale-like tubercles ellipsoided in a diagonal cross section on the skin surface of the body. And also this species moved like incessant wiggly movement pattern as that of fly maggot duing the larval development. As regards the tubercles and larval movement pattern, the larvae of A. asmussi shared similar characters with those of Acheilognathus rhombeus, A. longipinnis and Pseudoperilampus typos. Hence A. asmussi may be closely related to these species. The latter three species are autumn-spawning bitterlings and the larval development always retard in a certain stage, while A. asmussi is a spring-summer spawning species and the larval development never retards. Judging from these facts, it should be considered that there are close relations in evolutionary trend between the two spawning species.

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Kit Preparation and Biodistribution of $Bz-MAG_3$ (benzoylmercaptoacetylglycylglycylglycine) for Renal Imaging (신장기능영상용 방사성의 약품 $Bz-MAG_3$(Benzoylmercaptoacetylglycylglycylglycine) 의 키트화 및 체내분포)

  • Kim, Young-Ju;Jeong, Jae-Min;Cho, Jung-Hyuk;Chang, Young-Soo;Lee, Dong-Soo;Chung, June-Key;Lee, Myung-Chul;Koh, Chang-Soon
    • The Korean Journal of Nuclear Medicine
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    • v.30 no.3
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    • pp.367-371
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    • 1996
  • The $MAG_3$ is a tubular excreting radiopharmaceutical for renal image. We synthesized benzoyl $MAG_3\;(Bz-MAG_3)$ and made a kit for labeling with $^{99m}Tc$. We checked the labeling efficieny of $^{99m}Tc$ labeled $MAG_3$ and biodistribution. Labeling efficieny was checked by TLC-SG (acetonitrile/$H_2O$=2/1). After injecting of 1 mCi of $^{99m}Tc-MAG_3$ to ICR-mice, $T_{max}(min),\;T_{1/2}(min)$ were obtained in the renogram. Sequential images (30sec, 2min, 5min, 10min, 15min, 20min) of $^{99m}Tc-MAG_3$ were compared with those of commercial $^{99m}Tc$-DTPA (Du Pont Merck Pharmaceutical Co.) kit. 1) The $R_f$ value of synthesized $^{99m}Tc-MAG_3$ was 0.78 and labeling efficiency was $97.5{\pm}1.9%$ (n=10). 2) The dynamic images of the $^{99m}Tc-MAG_3$ were better than those of the $^{99m}Tc$-DTPA. 3) The $T_{max}(min.)$ and $T_{1/2}(min.)$ of $^{99m}Tc-MAG_3$ (n=10) were $1.5{\pm}0.5$ (left), $1.4{\pm}0.4$ (right), and $4.3{\pm}1.4$ (left), $4.8{\pm}2.0$ (right), respectively. The $T_{max}(min.)$ and $T_{1/2}(min.)$ of $^{99m}Tc$-DTPA (n=7) were $2.7{\pm}1.6$ (left), $2.7{\pm}1.6$ (right), and $3.8{\pm}1.7$ (left), $4.5{\pm}2.7$ (right), respectively. The quaility of image and labeling efficiency of the synthesized $Bz-MAG_3$ kit were excellent, that it was supposed to be used in routine clinical work.

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Effect of Pyroligneous Liquor on Oxygen Radicals and Their Scavenger Enzymes in Liver of CD Rats (흰쥐 간조직의 활성산소 및 제거효소에 미치는 목초액의 영향)

  • Choi, Jin-Ho;Cho, Weon-Ki
    • Journal of Nutrition and Health
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    • v.40 no.2
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    • pp.111-117
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    • 2007
  • This study was designed to investigate the effects of pyroligneous liquor on oxygen radicals and their scavenger enzymes in the liver of Cri/Bgi CD rats (7 rats per group). Male rats were fed a basic diet prepared in our Lab., PL-0 (Control), PL-1, PL-25, PL-50 and PL-75 groups were Prepared to be 0%, 1%, 25%, 50% and 75%with distilled water using pyroligneous liquor (35% of Choa Co. Ltd.), and were administrated orally for 8 weeks. Superoxide radical contents in liver mitochondria and microsomes were significantly decreased to 12-14%, 11-15%, respectively, in these PL-25 and PL-50groups compared with the control group. Hydroxyl radical content in mitochondria and microsomes were markedly decreased to 12-20% and 17%, respectively, in these PL-25 and PL-50% groups compared with the control group. Hydrogen peroxide content in mitochondria and microsomes were significantly decreased about 15-12% and 22-20% in liver of PL-25 and PL-50 groups compared with the control group. Mn-SOD and Cu/Zn-SOD activities in liver of PL-25 and PL-50 groups were remarkably increased to 15-25%, 11-16%, respectively, compared with the control group. GPx activities in mitochondria and microsomes were significantly increased in the liver of PL-25 and PL-50 groups compared with the control group. CAT activities in mitochondria and cytosol were significantly increased to 12-14%, 15-27%, respectively, in the liver of PL-25 and PL-50 groups compared with the control group. These results suggest that long term administration orally of 25 and 50% pyroligneous liquor may effectively inhibit the formation of oxygen free radicals, and also scavenger enzyme activities significantly increase through the administration orally.

Immunohistochemical Localization and the Characteristics of Antigenic Compnent Inducing IgE and IgG Antibodies in Spirometra erinacei (Spirometra erinacei에서 IgE와 IgG 항체를 유도하는 항원성분의 면역조직화학적 위치와 특성)

  • Chang-Hwan Kim;Sook-Jae Seo;Hong-Ja Kim;Kee-Hoon Kwak
    • Biomedical Science Letters
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    • v.2 no.1
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    • pp.1-12
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    • 1996
  • Antigenic components reacting with IgE and IgG antibodies were localized in muscular layer of adult and of larva, sparganum. But the antigenic components inducing IgG were localized at tegument and parenchyma in addition to muscular layer in adult and sparganum. Also in sparganum, the surface of calcareous corpuscles of parenchyma showed immunoreactivity to IgG antibody. However antigenic components inducing IgE antibody were not localized in tegument and parenchyma, but in adult worm, we observed the immunopositive reaction at the lining of vitelline follicles in mature proglottis and on surface of egg shell within uterus of graved proglottis. By the method of immunogold-labelling, we observed the location of antigenic particles in tegument of sparganum. The density of antigenic particles inducing IgG was higher than that of antigen particles inducing IgE in syncytial tegument, tegument cells. A total of 43 and 36 protein bands were resolved from crude extracts of adult and sparganum, respectively, by SDS-PAGE. 34 bands from crude extracts of adult and larva were migrated to same positions. By EITB, 21 bands of 44 bands in adult were recognized with IgG antibody, and also 21 bands of 36 bands in sparganum. 13 bands of them were common antigenic components both in the adult worm and sparganum. Because 19 bands of 44 bands in adult worm were reacted with IgE antibody, they were IgE antigenic component. In sparganum, 13 bands were IgE antigenic components. 9 bands of them were common antigenic component inducing IgE antibody in both a-dult and sparganum. 3 bands of antigenic component recognized by IgE and IgG antibody were nonspecific antigen in both adult and sparganum of Spirometra erinacei.

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Effects of the Exercise Training on Aging Heart in Rat I. Long Term Endurance Exercise (운동훈련이 흰쥐 노화심근에 미치는 영향 I. 장기간 지구력 운동 훈련)

  • 박원학;이상선;이용덕
    • Biomedical Science Letters
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    • v.2 no.1
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    • pp.71-90
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    • 1996
  • There is considerable current interest in the effect of regular vigorous exercise and in particular endurance-running as a possible measure in improving myocardial function. Some data indicate that the aging heart may actually suffer from vigorous endurance exercise. On the contrary appropriate exercise in aged animals improves myocardial function and aerobic energy metabolism. So far there is relatively little data to indicate that endurance exercise is in fact beneficial in improving myocardial function or damaging to heart of aged animals. The present investigation aimed to study the possible effect of a long range treadmill training program on the heart in aging rats. Male rats aged 3, 10, and 20 months were divided at random into a control (sedentary) and an exercise group. The training group was exercised for 5 days a week on an automated treadmill for 20minutes at 18m/min over a period of 5 months. The exercise regimen of our experiments did not cause any significant changes in the tissues and ultrastructural as com-pared with sedentary age-matched control. Tissues and ultrastructures of myocardial cells in trained group aged 8 months are intact and well organized as well as sedentary control group. Age associated tissue and ultrastructural changes of trained group aged 15 months included : an increase in transformed mitochondria, vacuoles, lysosomes, lipid droplets and early lipofuscin. But the trained heart did not show significant difference in tissue and ultrastructural properties from those of sedentary controls. Endurance-trained group aged 25 months showed significant qualitative tissue and ultrastructural difference as compared with age-matched controls. In addition to those found in 25 months control group, focal necrosis, myofibril fraying, hypercontraction band, seperation of intercalated discs, degenerating nucleus and infiltration of collagenous fiber into myocyte were noted in trained 25 months group. The stereological examination of the mi-crographs disclosed no significant difference in the myoflbril, mitochondrion, sarcotubule and in-terstitium volume density and surface density of mitochondrial cristae and numerical density of mitochondria between trained and control group aged 8 and 15 months. In the trained 25 months group, significant increase in volume density of interstitium, lipofucsin granule were shown as compared to untrained age-matched control. On the other hand, significant decrease in mitochondrion volume density was shown. The myofibril volume density did not differ between trained and control group although trained group showed slight increase. From the data obtained a reduced mitochondria/myofibrils ratio was found in trained rat heart aged 25 months and there was no difference between trained and control rat aged 15 months. But a slight but not significant increase was found in the trained group aged 8 months as compared with same age control group. Such increase in the ratio in young animals is considered to be of great importance to cardiac pumping and adaptability. Whereas such adaptations don't seem to occur in aged heart muscle. This study proposed that repeated endurance exercise do not cause any significant qualitative and quantitative ultrastructural change of heart muscle in young(3months) and adult (10months) suggesting that the heart is able to adapt to the exercise. On the contrary, the repeated endurance exercise stress may actually induce degenerative changes in the aged heart muscle(20months).

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Effect of Exercise Training on Aging Atrophyin Rat Skeletal Muscle III. Effect of Short Term Exercise Training for Senile Rat (흰쥐 골격근의 노화성 위축에 대한 운동훈련의 영향 III.노화 흰쥐에 적용한 단기간의 운동훈련의 영향)

  • 박승한;박원학;정형재
    • Biomedical Science Letters
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    • v.2 no.1
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    • pp.91-108
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    • 1996
  • The present study was designed to examine effect of short term treadmill and weight-training on aging arophy in the rat skeletal muscle. Male rats of 24 months old were used. Each groups included control, treadmill and weight-training for 4 weeks by using treadmill apparatus and body press apparatus. The histo and cytochemical, ultrastructural and stereological changes in senile skeletal muscles of the rat were observed in the present study. During the training period the body weight and muscular weight in all groups remained constant. The volume density of muscle fiber type IIC and IIB were increased, that of type IIA was decreased, but type I remained constant in treadmill-training group. In weight-training rat, the density of type IIA and IIB were increased, both those of type IIC was decreased. But, all changes of muscle fiber type is not significant. Senile control group some usual formation of mild contraction band, liposuscin pigment and muscular splitting were observed. After treadmill-training, histological and ultrastructural changes occurred in the muscle fiber, such as irregularity of the sarcolemma, interfibrillar vacuolization, longitudinal splitting, and widened I-bond. After weight-training, the changes occurred in the trained muscle fiber, such as appearances of many lysosomes and autophagic vacuoles, severe contraction band, and breakup of myofibrils. Histo and cytochemical studies showed that the activities of succinic dehydrogenase and acid phosphatase remained constant, activities of $Mg^{++}$-ATPase decrease with training. Stereological changes were not observed in the volume and numerical density of all subject component, but the surface density of mitochondrial inner membrane was increased with treadmill-training. These experimental results suggested that endurance training during short-term may result in the adaptible response in senile skeletal muscles. On the other side, weight-training is bad for senile skeletal muscle.

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