• Journal of Oral Medicine and Pain
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• 제24권4호
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• pp.397-410
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• 1999

In order to obtain the basic data concerning the optimal parameters in using Nd:YAG laser as a therapeutic modality to dentinal hypersensitivity, the author prepared 3 sections of sound dentin and 10 sections of sclerotic dentin with thickness of $0.5mm{\pm}0.1mm$ from human extracted teeth of anteriors and premolars, and applied the laser energy from a fiberoptic delivered, free running, pulsed Nd:YAG laser (wavelength 1064nm, pulse duration $120{\mu}sec$, fiber diameter $320{\mu}m$) to surfaces of sound and sclerotic dentin sections for 1 second with contact/unidirectional moving mode of the fiber under speed of 3mm~4mm/sec and parameters of 0.5W/10Hz, 1.0W/10Hz, 1.5W/10Hz, 2.0W/10Hz: $62J/cm^2$, $124J/cm^2$, $187J/cm^2$, $249J/cm^2$. The author comparatively evaluated the characteristics of ultrastructural changes on surfaces of sound and sclerotic dentin sections irradiated by the pulsed Nd:YAG laser using the scanning electron microscopy. A fairly ill-defined bordered surface of partially closed and melted dentinal tubules can be seen on the scanning electron microscopic feature of the sound dentin surface irradiated by the pulsed Nd:YAG laser with energy density of $62J/cm^2$. The physical modification of sound dentin surface extensively occurred depended on the increase of energy density from $62J/cm^2$ to $124J/cm^2$, $187J/cm^2$, $249J/cm^2$. While, a fairly well-defined bordered surface of partially closed and melted dentinal tubules with thickened peritubular dentin can be seen on the scanning electron microscopic feature of the sclerotic dentin surface irradiated by the pulsed Nd:YAG laser with energy density of $62J/cm^2$. The physical modification of sclerotic dentin surface of a fairly rough, shallow depression with many cracks, thickened peritubular dentin and structureless dentinal tubules extensively occurred depended on the increase of energy density from $62J/cm^2$ to $124J/cm^2$, $187J/cm^2$, $249J/cm^2$ compared to those of sound dentin surface irradiated by the pulsed Nd:YAG laser under the same parameters. Therefore, it is recommended that the pulsed Nd:YAG laser as a therapeutic modality to dentinal hypersensitivity should be applied with the less energy density than $62J/cm^2$ on the sound dentin surface, and its energy density on the partially sclerotic dentin surface should be lower than that on the sound dentin surface to preserve tooth from unnecessary excessive structural destruction.

• Restorative Dentistry and Endodontics
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• 제21권1호
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• pp.227-241
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• 1996

The effect of collagen dissolution in acid conditioned dentin was morphologically examined by both scanning and transmission electron microscopy. 18 freshly extracted human molars and dentin bonding systems of All Bond 2, Scotchbond Multipurpose, Superbond D-Liner were used in this study. For SEM preparation, each 3 of ~ exposed dentin surfaces were acid conditioned by using various acids within the above three bonding systems respectively. After acid conditioning of the other 3 exposed dentin surfaces as above, they were treated with 1.7% NaOCl for 2 minutes. The remaining 3 dentin surfaces were acid conditioned and treated with 3.3 % NaOCl for 2 minutes. All of the specimens were then fixed in 4 % glutaraldehyde for 12 h at $4^{\circ}C$ and dehydrated in ethanols grades from 50 % to 100 %, then surface changes of the specimens were observed by using SEM. For TEM preparation, exposed dentin surfaces were acid conditioned with the same acid as SEM specimens and treated with 1.7%, 3.3 % NaOCl respectively, then applied with corresponding bonding agents. After the procedures were finished, composite resin were applied on the dentin surfaces and light cured. Small, rectangular sticks with end dimensions of approximately 1 by 1 mm were sectioned and further sample preparative techniques for transmission electron microscopy were performed in accordance with the procedures used for ultrastructural TEM observations of calcified tissues. The results were as follows : 1. In the 1.7 % NaOCl retreated specimens after acid conditioning, the porous dentin surface of intertubular dentin and wide opening of dentinal tubules were appeared. And there were fine irregularities on the intertubular dentin, indicating a clear difference as compared with the acid conditioned specimens. 2. In the 3.3% NaOCl retreated specimens after acid conditioning, the intertubular dentin was further eroded causing a more porous and wider opening of dentinal tubules. Moreover, sharp irregularities on the intertubular dentin were more evident than those of acid conditioned and 1.7% NaOCl retreated specimens. 3. In all of the acid conditioned specimens, the resin-dentin hybrid layer of approximately 3.5mm thickness was formed and the collapsed collagen layer was observed on the uppermost part of hybrid layer in the specimens applied with All Bond 2. The collgen fibrils of intertubular dentin in specimens applied with Scotchbond Multipurpose were running perpendicular to the interface, and electron dense black layer demarcated from the deep unaltered dentin was more evident in the specimen applied with Superbond D-Liner than any other specimens. 4. In the 1.7 % NaOCl retreated specimens after acid conditioning, the resin-dentin hybrid layer of approximately 2.5-3.0mm thickness was formed and the collapsed collagen layer and longitudinally running collagen fibrils as shown in the acid conditioned specimens were observed in the specimens applied with All Bond 2 and Superbond D-Liner. 5. In all of the 3.3% NaOCl retreated specimens after acid conditioning, the evidence of resin-dentin hybrid layer was not identified ; nevertheless, the longitudinally running collagen fibrils remained slightly in the specimens applied with All Bond 2.

• 한국동물학회지
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• 제31권4호
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• pp.273-282
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• 1988

생쥐에 PMSG와 hOG를 주사한 후 난자-난구복합체의 미세구조의 변화를 환찰함으로세 난구세포의 분산현상을 규명하고자 본 실험을 행하였다. 난자는 PMSG 주사후 48시간까지 별 다른 변화가 없었고 다만 표면막에 miGrOVilli와 Coaled pit의 수가 감소하는 경향을 보였다. 그러나 PMSG-hCG주사 12시간 후에 배란된 난자의 표면은 microvilli와 coated pit가 사라져서 평평하게 되었다. 방사관세포는 PMSG주사 48시간 후메 밀착해 있던 투명대와 간격이 생기기 시작하였고, 투명대를 통관하여 난자의 표면막과 desmosome으로 연결되어 있던 세포질돌기도 퇴화의 징후를 보였다. PMSG-hCG주사 후에는 급속히 격리, 분산되고 세포질돌기는 퇴화하였으며 dermo-some도 사라겼다. 난구세포들은 대조군에서 밀집되어 있었고 거의 gap junction으로 연결되어 있었는데, PMSG주사 24시간 후에는 모양이 등글게 되고 더욱 밀집되었으며, 48시간 후에는 거의 loose junction으로 연결되었고 분산되기 시작하였다. 결국 PMSG-hCG주사 If시간 후에는 완전히 분산되었고 거의 모두 핵응축과 괴사현상을 보였다. 난자- 난구 복합체의 분산은 배란전에 PMSG에 의하여 시작되고 hCG에 의하여 촉진 완결된다는 것이 확실하다. The ultrastructural changes of the oocyte-cumulus complexes of mouse alter injection of PMSG and hOG have been investigated in order to elucidate expansion phenomenon of the cumulus cells. The oocytes until 48 hours after PMSC injection showed no change except a tendency of decrease in numbers of microvilli and the coated pelts on surface membrane. However, surface membrane of the ovulated oocytes 12 hours after PMSC-hCC injection changed to be smooth due to disapperance of microvilli and coated pits. Corona radiate cells tightly attaching to zona pe]lucida 48 hours after PMSC injection began to be detached and their cytoplasmic processes connected by desmosome to oocyte surface membrane showed a degeneration symptom. Thereafter the detachment and degeneration were accelerated by hCG injection and followed by disappearence of desmosome. The cumulus cells in control group were compacted and connected by almost 9aP junction each another. Ite cumulus cells 24 hours after PMSG injection were changed to be round form and more tightly compacted. However, the cumulus cells 48 hours after PMSG injection were connected by almost loose junction and showed the beginning of expansion. Eventuallv, the cumulus cells 12 hours a%or PMSG-hCG injection were completely expanded, and became pvknotic and necrotic in most It is clear that the expansion of oocyte-cumulus complex were initiated by PMSC, then accelerated and completed by hCG before ovulation.

• 대한소아치과학회지
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• 제24권3호
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• pp.660-677
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• 1997

The purpose of this study was to examine the morphological characteristics and combined effects of fluoride application and laser irradiation on artificial caries-like lesion formation in bovine enamel. Enamel specimens were divided into five experiment group and placed in no-treatment(group C), APF application alone(group F), laser irradiation alone(group L), APF application before laser irradiation (group FL), and APF application after laser irradiation(group LF) on artificial caries-like lesion. Sound enamel was used as a control group. The ultrastructural changes and physical effects of the fluorided and lased enamel has been investigated by using SEM, enamel solubility and microhardness test as well as distributions of calcium, phosphorus and fluoride in internal enamel by using EPMA. The following results were obtained. 1. In the all experiment groups, the amounts of dissolved calcium of enamel surfaces significantly decreased according to increasing exposure time of acid solution than control group(P<0.001). Group L showed higher than that group FL and LF in 30 and 60min(P<0.05). 2. The microhardness values of enamel surface in the control group was highest than that in the other experiment groups. Group F, L, FL and LF were significantly increased than group C(P<0.001). The enamel surface treated with APF produced deposites of numerous small globules and lased enamel showed a cracker-like appearance with microcrack and small pore. Numerous deposits were infiltrated in the fissured portion of enamel treated with APF after laser irradiation. 4. In the case of APF application alone, the elevation of the fluoride profile can be seen within $5{\mu}m$ of the outermost layer and a similar profile observed in the specimen treated with APF before laser irradiation. However, the specimen treated with APF after laser Irradiation showed a large elevation within $10{\mu}m$ of the outermost layer of the enamel. 5. The higher Ca/P ratios were observed in $10{\mu}m$ depth of lased and fluorided enamel when compared to the sound and carious enamel. The fluoride content decreased rapidly with distance from enamel surface, in the group F, fluoride concentration was significantly higher than that in the group C, L, FL, LF and control group according to increasing enamel depth (P<0.05).

• Asian Pacific Journal of Cancer Prevention
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• 제13권12호
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• pp.6191-6196
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• 2012

Aims and Background: Traditional chemotherapy strategies for human leukemia commonly use drugs based on cytotoxicity to eradicate cancer cells. One predicament is that substantial damage to normal tissues is likely to occur in the course of standard treatments. Obviously, it is urgent to explore therapies that can effectively eliminate malignant cells without affecting normal cells. Our previous studies indicated that ginsenoside $Rg_1$ ($Rg_1$), a major active pharmacological ingredient of ginseng, could delay normal hematopoietic stem cell senescence. However, whether $Rg_1$ can induce cancer cell senescence is still unclear. Methods: In the current study, human leukemia K562 cells were subjected to $Rg_1$ exposure. The optimal drug concentration and duration with K562 cells was obtained by MTT colorimetric test. Effects of $Rg_1$ on cell cycle were analyzed using flow cytometry and by SA-${\beta}$-Gal staining. Colony-forming ability was measured by colony-assay. Telomere lengths were assessed by Southern blotting and expression of senescence-associated proteins P21, P16 and RB by Western blotting. Ultrastructural morphology changes were observed by transmission electron microscopy. Results: K562 cells demonstrated a maximum proliferation inhibition rate with an $Rg_1$ concentration of $20{\mu}\;mol{\cdot}L^{-1}$ for 48h, the cells exhibiting dramatic morphological alterations including an enlarged and flat cellular morphology, larger mitochondria and increased number of lysosomes. Senescence associated-${\beta}$-galactosidase (SA-${\beta}$-Gal) activity was increased. K562 cells also had decreased ability for colony formation, and shortened telomere length as well as reduction of proliferating potential and arrestin $G_2$/M phase after $Rg_1$ interaction. The senescence associated proteins P21, P16 and RB were significantly up-regulated. Conclusion: Ginsenoside $Rg_1$ can induce a state of senescence in human leukemia K562 cells, which is associated with p21-Rb and p16-Rb pathways.

• Applied Microscopy
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• 제17권1호
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• pp.47-64
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• 1987

Morphological difference of the renal glomerulus at different age groups have been studied in young (three month-old), adult (twelve month-old) and old (thirty month-old) Fisher strain 344 rats. Pieces of the tissues were taken from renal corticies prefixed with 2.5% glutaraldehyde-1.5% paraformaldehyde (0.1 M Millonig's phosphate buffer, pH 7.3), following by post-fixation with 1% osmium tetroxide (0.1 M Millonig's phosphate buffer, pH 7.3) and embedded within Araldite. The ultrathin sections contrasted with uranyl acetate and lead citrate were observed under a JEM 100CX electron microscope. The mean thickness of glomerular basal lamina and Bowman's capsule were determined by measuring the thinnest portion of basal lamina, and by taking the average of 50 readings from electron micrographs at different ages. The numerical changes of the slit pores were compared based upon the numbers over the length of 10um of glomerular basal lamina. The results were as follow: 1. The thickness of glomerular basal lamina is increased during aging; 140.4 nm in young rats, 270.0 nm in adult ones, and 437.8 nm in old ones. 2. The thickness of basal lamina of parietal cells of Bowman's capsule is 187.5 nm in young rats, 914.0 nm in adult ones, and 2850.0 nm in old ones. 3. The numbers of the slit pores of basal lamina are reduced during aging, 30.3 slit pores/$10{\mu}m$ in adult ones, and 24.2 slit pores/$10{\mu}m$ in old ones. 4. Accumulation of dense intracytoplasmic filamentous material in the parietal cells of Bowman's capsule is increased in the vicinity of the basal lamina during aging. The proximal tubule-like epithelial cell in Bowman's capsule is observed at one glomerulus in a young rat. 5. The endothelial cells are edematous and form balloon-like structure protruding into capillary lumen in young and old rats. 6. Cytoplasm of the podocyte shows a variety of alteration during aging, such as swelling of mitochondria and of endoplasmic reticulum, and increase of microtubules, microfilaments, lysosomes and lamellated myelin structures, etc. Accumulation of dense intracytoplasmic material in the foot processes is increased in the vicinity of the basal lamina during aging. The podocytic membrane-like structures are seen in young and o]d rats. 7. The mesangial matrices and mesangial cells are increased during aging, and slight swelling of endoplasmic reticulum and Golgi cisternae in young and old rats.

• Applied Microscopy
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• 제31권3호
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• pp.275-285
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• 2001

폐포강 대식세포는 사이토카인, 유해산소 대사물을 포함한 그들이 분비하는 물질들로 인해 급성 폐손상에 있어서 직접, 간접적으로 폐손상의 초기반응에 중요한 역할을 담당하는 것으로 알려져 있다. 본 연구에서는 $interleukin-1\alpha$(IL-1)로 유도된 급성 폐손상에서 폐포강 대식세포의 역할을 알아보고자 하였다. 실험군은 대조군과 IL-1투여 후 1시간, 2시간, 3시간, 4시간 그리고 5시간군으로 나누었으며, 폐포강 대식세포와 XO와의 관계를 분석하기 위해 폐세척액 내 XO의 활성도와 폐포강 대식세포, 단핵구, 그리고 호중구의 수적 변화를 측정하였다. 그리고 각 군의 미세구조 변화를 관찰하였다. 실험 결과, 폐포강 내 단핵구의 수는 IL-1투여 후 1시간군에서 대조군과 비교하여 현저히 증가되었으며 (p<0.001), 폐포강 대식세포의 수는 IL-1 투여 2시간 후에 가장 높았고, 폐세척액 내 XO의 활성도는 IL-1 투여 후 점차적으로 증가되다가 3시간 후에 현저히 증가되었다(p<0.05). 폐포강 내 호중구의 수는 IL-1투여 3시간 후부터 뚜렷이 증가되기 시작하였다. 이러한 결과로 보아 IL-1을 기관지 내로 투여한 후 유도된 급성 폐손상에서 폐포강 대식세포에서 유리된 XO는 호중구의 축적에 의한 손상보다 더 초기단계에서 폐손상을 유도하는 인자인 것으로 추정된다.

• Applied Microscopy
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• 제34권3호
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• pp.185-197
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• 2004

본 연구의 목적은 납중독에 의한 신장 기능 손상에 대한 스쿠알렌의 효과를 관찰하고자 하였다. ICR계 건강한 생쥐를 사용하여 납과 스쿠알렌을 복강 투여한 후, NO와 조직학적 변화를 관찰하였다. 실험군 설정은 다음과 같다. 실험군 1은 정상군, 실험군 2는 납 (30 mg/kg)만 처치한 군, 실험군 3은 납을 처치하고 스쿠알렌 (180 mg/kg)도 함께 처치한 군으로 각 실험군 당 생쥐 10마리를 사용하였다. 결과는 다음과 같다. 실험군 2의 경우, 미토콘드리아 신장, 사립체내막의 파괴, 소포체에서 리보소옴의 탈락이 24, 48시간동안 관찰되었고, 72시간부터 서서히 회복 되는 것을 관찰할 수 있었다. 실험군 3의 경우 실험군 2에 비해 손상 정도가 덜 하였으며, 48시간 이후부터 정상군과 유사한 소견을 보였다. NO의 경우 실험군 2에서는 NO 수치가 감소하였다. 하지만, 실험군 3의 경우 정상군보다 NO가 증가하였을 뿐만 아니라 납에 의한 NO 수치 감소에 대해 회복 효과를 보였다. 위와 같은 결과로 보아, 스쿠알렌이 신장 근위세뇨관에 미치는 납 독성을 감소시키면서 빠른 회복에 효과가 있을 것으로 사료된다.

• Applied Microscopy
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• 제37권4호
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• pp.249-258
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• 2007

이자섬은 이자를 구성하는 외분비조직에 둘러싸여 존재하는 내분비세포의 집단으로, 이자섬에서 분비되는 인슐린은 $\beta$세포에서 분비되는 호르몬이며, 세포질의 리보좀에서 합성되고 골지체를 경유하여 세포질로 방출되는 기작을 가지고 있다. 충분한 양의 이자섬 이식은 인슐린 의존형 당뇨병인 제1형 당뇨병에서 정상혈당을 회복시키고, 당뇨 합병증을 방지할 수 있는 치료방법으로 사용되고 있다. 하지만 당뇨병 환자에게 이식을 위한 이자섬의 양에 비해 공여자로부터 증여된 이자섬의 양은 제한적이다. 이러한 문제점은 이자섬의 증식으로 연구되고 있으나, 배양된 이자섬이 정상 조직내의 이자섬과 형태적 기능적으로 동일한 것인지에 관한 연구는 미비하였다. 따라서 본 연구에서는 분리된 이자섬과 배양된 이자섬을 구성하는 세포들의 내부구조의 변화를 주사전자현미경, 투과전자현미경을 이용하여 세포의 미세구조를 확인하고, 인슐린 항체를 이용한 $\beta$세포 내의 인슐린 분포양상을 확인하여 다음과 같은 결과를 얻었다. 분리된 이자섬의 $\beta$세포는 일반적인 핵 미토콘드리아, 세포질세망 그리고 인슐린 과립이 분포하고, 배양된 이자섬 $\beta$세포의 경우 분리된 이자섬에 비하여 일반적인 핵의 모습과 부피가 증가한 세포질과 미토콘드리아, 세포질세망 그리고 골지체의 발달이 이루어지는 것으로 관찰되었다. 인슐린 과립의 경우 분리된 이자섬에 비해 감소하며, 세포막 주위에 분포하는 것으로 관찰되었다. 배양된 이자섬에서 관찰되는 인슐린 과립 분포의 변화, 세포질세망의 증가, 골지체의 발달은 배양된 이자섬 $\beta$세포의 인슐린 생성 분비 기능의 향상과 부피의 증가가 이루어지기 위한 세포 내부의 형태적 변화가 이루어지는 것으로 추측된다.

• Applied Microscopy
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• 제38권1호
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• pp.1-10
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• 2008

본 연구는 급성 수은 독성에 대한 민들레의 효과를 알아보기 위하여 시도되었다. 30 g 내외의 생쥐를 대상으로 대조군, 수은(5 mg/kg) 투여군, 수은 투여 후 민들레(3 g/kg)를 구강투여 한 후 다시 24, 48, 72, 96시간, 1주일군으로 세분하여 간장 손상 억제효과를 알아보기 위하여 생화학적 및 조직학적 실험을 실시하였다. 혈액중 aspartate amiotransferase (AST)와 alanine aminotransferase (ALT)의 수치는 민들레 투여군이 수은 투여군보다 감소되었다. 또한 간조직의 SOD와 catalase활성도 역시 민들레 투여군이 수은 투여군에 비하여 감소하였으나 통계학적 유의성은 없었다. 간 조직의 광학현미경적 관찰에서 심한 조직괴사가 관찰되지만 민들레 투여군에서는 문맥주위의 약간의 괴사와 심한 호중구 침윤현상이 관찰되었다. 전자현미경적 관찰 결과, 간장에서 수은 투여군은 간세포의 핵이 함입되어 불규칙했으며 미토콘드리아와 조면소포체의 수조가 팽대되고, 리보솜의 탈락이 관찰되었다. 민들레 투여군은 핵이 정상적인 상태로 관찰되었고, 전자밀도가 높은 미토콘드리아가 분포되어 있었으며, 리보솜이 부착된 상태로 층판구조를 형성하는 조면소포체가 관찰되었다. 이상의 연구 결과로 보아 민들레가 수은으로 유발된 간손상을 보호하는 효과가 있는 것으로 사료되며 민들레에 대한 다양한 연구가 필요할 것으로 사료된다.