• Title/Summary/Keyword: H-E stain

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EFFECTS OF COMPRESSION ON DISTRACTED AREA DURING MANDIBULAR DISTRACTION IN RABBITS (가토에서 하악골신장술시 신장부에 대한 가골압축의 효과)

  • Park, Hong-Ju;An, Jin-Suk;Kook, Min-Suk;Oh, Hee-Kyun;Ryu, Sun-Youl;Cho, Jin-Hyoung
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.29 no.1
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    • pp.1-9
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    • 2007
  • Purpose: To evaluate the effect of compression on the distracted area in new bone formation during mandibular distraction osteogenesis in rabbits. Materials and method: Sixteen rabbits, weighing approximately 2 Kg, and the prefabricated distraction device were used. With the rabbits under general anesthesia, we performed vertical osteotomies between the anterior and posterior mandibular teeth and then placed the distraction device. After a 5 -day latency period, the mandible was distracted to a length of 10.0 mm at a rate of 1 mm/day and then immediately compressed 4 mm in the experimental group (n=8). In the control group (n=8), the mandible was distracted to a length of 6.0 mm at a rate of 1 mm/day. Rabbits in the control group were killed at 2 and 8 weeks during the consolidation period. The specimens were evaluated with light microscope after H & E stain. Histomorphometric analysis was done at 8 week specimens. Results: All experimental animals showed mandibular elongation on the macroscopical and radiographic evaluations. At 2 week, immature bone formation was observed from the surface of the host bone margins with collagen fibers arranged parallel to the direction of distraction in the control group; in the experimental group, immature bone formation was observed adjacent to the host bone, and the collagen fibers were not arranged uniformly. At 8 week, spindle-shaped new bone formation was seen in the direction of distraction in distracted area of the control group, while in the experimental group, the newly formed bone was arranged in a multidirectional manner, like the pattern of trabeculae. In the histomorphometric analysis of 8 weeks, the area of bone deposition was $2.12{\pm}\;0.75\;inch^2$ in the experimental group and $0.87{\pm}0.51\;inch^2$ in the control group (p<0.01). The bone deposition ratio was $29.60{\pm}10.50%$ in the experimental group and $12.10{\pm}7.17%$ in the control group (p<0.01). Conclusion: These results suggest that compression after over-distraction during the mandibular distraction osteogenesis is an effective method of increasing the amount of newly formed bone in distracted area.

Characteristics of Wound Contraction according to the Shape and Antomical Regions of the Wound in Porcine Model (돼지모델에서 상처의 모양과 부위에 따른 상처수축의 특성)

  • Chu, Ho-Jun;Son, Dae-Gu;Kwon, Sun-Young;Kim, Jun-Hyung;Han, Ki-Hwan
    • Archives of Plastic Surgery
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    • v.38 no.5
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    • pp.576-584
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    • 2011
  • Purpose: The shape and location, the amount of the wound and the characteristics of the remaining tissues are known to influence wound contraction. The previous studies using small animals have not been an appropriate model because the wound healing mechanisms and skin structures are different from those of the human. The purpose of this study is to evaluate wound contraction according to the shape and location of the wound using a $Micropig^{(R)}$. Methods: Four $Micropigs^{(R)}$ (Medikinetics, Pyeongtaek, Korea) that were 10 months of age weighed 25 kg were used. Full thickness skin defects were made by clearing all the tissues above the fascial layer in the shape of square, a regular triangle and a circle of 9 $cm^2$ each on the back around the spine. Eight wounds were created on the back of each pig, 50 mm apart from each other. The randomly chosen wound shapes included 11 squares, 11 regular triangles, and 10 circles. Wound dressing was done every other day with polyurethane foam. The wound size was measured using a Visitrak $Digital^{(R)}$ (Smith & Nephew, Hull, UK) on every other day after surgery from day 2 to day 28. A biopsy was performed on day 3, and 1, 2, 3 and 4 weeks to investigate the degree of acute and chronic inflammation, the number of microvesssel and myofibroblast density using H & E stain and immunohistochemistry. The wound contraction rate was calculated to figure out the differences among each of the shapes and the locations. Results: The ultimate shape of the circle wound was oval, and that of the regular triangle and square were stellate. The maximum contraction rate was obtained on 8 to 10 days for all the shapes, which corresponds with the immunohistochemical finding that myofibroblast increases in the earlier 2 weeks whereas it decreases in the later 2 weeks. Epithelialization was seen in the wound margin on day 7 and afterwards. The final wound contraction rates were highest for the regular triangle shapes; however, there were no statistically significant differences. The wound contraction rates by locations showed statistically significant differences. The wound in the cephalic area presented more contractions than that of the wounds in the caudal area. Conclusion: The location of a wound is more important factor than the wound shape in wound contraction.

Anti-diabetic effects of the extract from Atractylodes lancea, Anemarrhena asphodeloides and Cinnamomum Cassia mixture in high fat diet-induced diabetic mice and regulation of the function in C2C12 mouse skeletal muscle cells (창출·지모·육계 복합추출물의 고지방식이 유도 당뇨병 마우스에서의 항당뇨 효능 및 C2C12 골격근세포에서의 조절기전 연구)

  • Park, Ki Ho;Kang, Seok Yong;Kang, Anna;Jung, Hyo Won;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.34 no.6
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    • pp.79-89
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    • 2019
  • Objective : This study investigated the anti-diabetic effects of DM1, a herbal mixture with Atractylodis Rhizoma, Anemarrhenae Rhizoma, and Cinnamomi Cortex in high fat diet (HFD)-induced diabetic mice and the mechanism in C2C12 mouse skeletal muscle cells. Methods : The C57B/6 mice were fed high fat for 12 weeks, and then administrated DM1 extract (500 mg/kg, p.o.) for 4 weeks. The changes of body weight, calorie and water intakes, fasting blood glucose levels and the serum levels of glucose, insulin, triglyceride, HDL-cholesterol, AST and ALT were measured in mice. The histological changes of liver and pancreas tissues were also observed by H&E stain. C2C12 myoblasts were differentiated into myotubes and then treated with DM1 extract (0.5, 1, and 2 mg/㎖) for 24 hr. The expression of myosin heavy chain (MHC), PGC1α, Sirt1 and NRF1, and the AMPK phosphorylation were determined in the myotubes by western blot, respectively. Results : The DM1 extract administration significantly decreased the calorie and water intakes, glucose, triglyceride, AST and ALT levels and increased insulin and HDL-cholesterol in HFD-induced diabetic mice. DM1 extract inhibited lipid accumulation in liver tissue and improved glucose tolerance. In C2C12 myotubes, DM1 treatment increased the expression of MHC, PGC1α, Sirt-1, NRF-1 and the AMPK phosphorylation. Conclusion : In our results indicate that DM1 can improve diabetic symptoms by decreasing the obesity, glucose tolerance and fatty liver in HFD-induced diabetic mice, and responsible mechanism is might be related with energy enhancement.

Studies on the MC1R Gene Frequencies in Landrace, Large White, Duroc and Jeju Native Black Pigs (랜드레이스, 대요크셔, 듀록 및 제주 흑돈의 Melanocortin 1 Receptor(MC1R) 유전자의 유전자형 분석)

  • Cho, I.C.;Lee, J.G.;Jung, J.G.;Yang, B.S.;Kang, S.Y.;Kim, B.W.
    • Journal of Animal Science and Technology
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    • v.44 no.2
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    • pp.207-212
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    • 2002
  • This study was conducted to investigate the genotypes and frequencies of Melanocortin 1 Receptor(MC1R) genes in pigs which plays a central role in regulation of eumelanin (black/brown) and phaeomelanin(red/yellow) pigment synthesis within the mammalian melanocytes. Four different breeds of pigs(20 Landrace, 20 Yorkshire, 20 Duroc, and 93 Jeju native black pigs) were used and PCR-RFLP analysis of MC1R gene was also carried out. Two regions of MC1R genes (428bp and 405bp) were amplified using two specific primers (MERL1-EPIG2, EPIG1-EPIG3), respectively and MC1R allele were determined using 2 restriction enzymes (BspHⅠ, AccⅡ). The results of this experiment indicated that MC1R allelic type in Landrace, Large Yorkshire and Duroc were MC1R *2 (Ep), MC1R *2 (Ep), MC1R *4 (e), respectively. However, various allelic types of MC1R genes were detected in Jeju native black pigs. MC1R allelic type of Jeju black pigs was MC1R*2 type as in Meishan and Large black breeds or MC1R*3 type as in Hampshire and Berkshire breeds and the gene frequencies of ED1 and ED2 were 0.554 and 0.446 in average.

A STUDY ON THE DISTRIBUTION OF SEVERAL GROWTH FACTORS IN THE ARTIFICIALLY CHEATED CLEFT LIP WOUND HEALING OF RABBIT FETUSES (토끼 태자에 형성시킨 구순열상의 치유과정에서 수종 성장인자 분포에 관한 연구)

  • Baek, Seung-Hak;Yang, Won-Sik
    • The korean journal of orthodontics
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    • v.27 no.5 s.64
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    • pp.683-696
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    • 1997
  • Recently several growth factors such as $TGF-{\beta}1$, $TGF-{\beta}1$, PDGF, bFGF are known to play an important role in scar formation following adult tissue injury. But there is little known about the role of growth factors in fetal tissue healing without scar formation. Therefore the purpose of this study is to investigate the distribution of growth factors which we involved with scar formation in the artificially created cleft lip wound healing of fetuses. The author had undergone hysterotomy and created cleft lip-like defects on fetuses of New Zealand White Rabbit in mid-third trimester (24 days). Fetuses were divided into 3 groups (the repaired group, the unrepaired group and the sham-operated control group). At 1, 2, 3, 5, 7 days after procedure, the repaired, the unrepaired and the control groups were obtained by Caeserean section. After documenting the viability of fetuses, fetuses were photographed to compare size and facial morphology and sectioned for histological examination by H & E stain and spatial and temporal deposition of $TGF-{\beta}1$, $TGF-{\beta}2$, PDGF, bFGF by immunohistochemical method. The findings are summarized as follows 1. There were lack of inflammation and scar formation and neovascularity in the repaired and the unrepaired group during experimental periods. 2. The reepithelialization of the unrepaired group was slower than that of repaired group. 3. There were no differences of distribution of bFGF in the control, the repaired and the unrepaired group. 4. PDGF was increased at post-op. first and second day and decreased after post-op. third day. Eventually, there were no differences in the control, the repaired and the unrepaired group. 5. $TGF-{\beta}1$ and $TGF-{\beta}2$ were slightly increased at post-op. first and second day and decreased after post-op. third day. Eventually there were no differences in the control, the repaired and the unrepaired group. And $TGF-{\beta}2$ is more densely stained than $TGF-{\beta}1$.

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A STUDY ON THE EXTRACELLULAR MATRIX IN THE ARTIFICIALLY CHEATED CLEFT LIP WOUND HEALING OF RABBIT FETUSES (토끼 태자에 형성시킨 구순열상의 치유과정에서 세포외기질 분포에 관한 연구)

  • Yang, Won-Sik;Baek, Seung-Hak
    • The korean journal of orthodontics
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    • v.28 no.1 s.66
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    • pp.1-15
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    • 1998
  • Adult wound healing is accompanied with inflammation and eventual scar formation, whereas fetal wounds heal rapidly by mesenchymal proliferation without significant inflammatory cell participation and with minimal or no scar formation. The cellular mechanisms underlying these differing forms of wound healing are unknown but the extracellualr matrix through its effects on cell function, may play a key role. Therefore the purpose of this study is to investigate the spatial and temporal deposition of several component of extracellular matrix, which are known to be involved with scar formation, in the artificially created cleft lip wound healing of fetuses. The author had undergone hysterotomy and created cleft lip-like defects on fetuses of New Zealand White Rabbit in mid-third trimester(24 days). Fetuses were divided into the repaired group, the unrepaired group and the sham-operated control group. At 1, 2, 3, 5, 7 days after procedure, fetuses were obtained by Caeserem section. After documenting the viability of fetuses, they were photographed to compare size and facial morphology and sectioned for histological examination by H & E stain and spatial and temporal deposition of collagen typeI, III, IV, V and fibronectit laminin by immunohistochemical method. The findings are summarized as follows 1. There were lack of inflammation in the repaired and the unrepaired group during experimental periods. 2. The reepithelialization of the unrepaired group was slower than that of the repaired group. 3. Collagen I, III, V were found from post-op. third day. There were no difference of distribution in the control, the repaired and the unrepaired group. Collagen types I, III, V were present in all groups with restoration of the normal collagen pattern in the fetus. This implies that lack of scarring in fetal wounds is due to the difference of collagen organization pattern within wound and not simply lack of collagen formation. 4. Collagen IV was slightly increased at post-op. third day and decreased after post-op. fifth day. Eventually there were no differences in the control, the repaired and the unrepaired group. Lminin was found at post-op. fifth day and maintained staining density until post-op. seventh day. There were no differences in the control, the repaired and the unrepaired group. According to staining of laminin and collagen type IV in epithelial basement membrane, formation of epithelial basement membrane was not completed until reepithelialization was finished. 5. According to staining of laminin and collagen type IV, there were no increase of neovascularity in the repaired and the unrepaired group. 6. Fibronectin was increased until post-op. third day at fibrin clot, wound base and margin and decreased after post-op. fifth day. Eventually, there were no differences in the control, the repaired and the unrepaired group. So it implies fibronectin plays a role as provisional matrix for fetal wound healing.

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Effect of Radiation Dose for Radiotherapy on Ovarian Follicle Atresia in Rat (치료 방사선량이 쥐의 난포 퇴축에 미치는 영향)

  • Lee, Won-Jeong;Seon, Jong-Ryul;Yoo, Se-Jong;Ahn, Bong-Seon
    • Journal of Radiation Protection and Research
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    • v.37 no.4
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    • pp.208-212
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    • 2012
  • In previous studies, ovarian follicle in rat has been used a higher radiation dose than that for cancer radiotherapy in clinical practice. The aim of this study was to evaluate the effect of radiation dose used for cancer radiotherapy on ovarian follicle atresia in rat. Mice of 4-week-old female were whole body irradiated with 2 cGy or 2 Gy (Mevatron 67, Siemens, Germany) and sacrificed by cervical dislocation. Ovaries were collected at 24 hours after irradiation to observe the degree of follicular atresia. Ovaries were fixed in neutral formaldehyde solution for 24 hours and embedded with paraffin. Cutted in $5{\mu}m$ thickness with microtome and stained with hematoxylin and eosin (H&E) and TUNEL immunohistochemical stain, and examined histologically under a light microscope. All data were presented as mean ${\pm}SD$, calculating the ratio of normal or atretic follicles to total ovarian follicles. Statistical analysis was performed by the Mann Whitney test using the SPSS ver 19.0. Ratio of atretic to total follicles of 2 Gy group was significantly higher than control or 2 cGy groups (p<0.05). Ratio of normal to total follicles of 2 Gy group was significantly lower than control group in preantral follicle (64.0 vs. 87.7, p=0.027). Ratio of normal to total follicles of 2 cGy group was significantly increased more than control or 2 Gy groups in antral follicle, and there were no significant difference between control and 2 Gy groups (p=0.522). Radiation dose of 2 Gy for cancer radiotherapy have a significant effect on ovarian follicle atresia in rat.

The neuroprotective effect of mycophenolic acid via anti-apoptosis in perinatal hypoxic-ischemic brain injury (주산기 저산소성 허혈성 뇌손상에서 항세포자멸사를 통한 mycophenolic acid의 신경보호 효과)

  • Kim, Ji Young;Yang, Seung Ho;Cha, Sun Hwa;Kim, Ji Yeun;Jang, Young Chae;Park, Kwan Kyu;Kim, Jin Kyung;Chung, Hai Lee;Seo, Eok Su;Kim, Woo Taek
    • Clinical and Experimental Pediatrics
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    • v.50 no.7
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    • pp.686-693
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    • 2007
  • Purpose : Mycophenolic acid (MPA), the active metabolite of mycophenolate mofetil (MMF), is a potent inhibitor of inosine-monophosphate dehydrogenase (IMPDH), a new immunosuppressive drug used. It was reported that MPA protected neurons after excitotoxic injury, induced apoptosis in microglial cells. However, the effects of MPA on hypoxic-ischemic (HI) brain injury has not been yet evaluated. Therefore, we examined whether MPA could be neuroprotective in perinatal HI brain injury using Rice-Vannucci model (in vivo) and in rat brain cortical cell culture induced by hypoxia (in vitro). Methods : Cortical cells were cultured using a 18-day-pregnant Sprague-Dawley (SD) rats and incubated in 1% $O_2$ incubator for hypoxia. MPA ($10{\mu}g/mL$) before or after a HI insult was treated. Seven-day-old SD rat pups were subjected to left carotid occlusion followed by 2 hours of hypoxic exposure (8% $O_2$). MPA (10 mg/kg) before or after a HI insult were administrated intraperitoneally. Apoptosis was measured using western blot and real-time PCR for Bcl-2, Bax, caspase-3. Results : H&E stain revealed increased brain volume in the MPA-treated group in vivo animal model of neonatal HI brain injury. Western blot and real-time PCR showed the expression of caspase-3 and Bax/Bcl-2 were decreased in the MPA-treated group In in vitro and in vivo model of perinatal HI brain injury, Conclusion : These results may suggest that the administration of MPA before HI insult could significantly protect against perinatal HI brain injury via anti-apoptotic mechanisms, which offers the possibility of MPA application for the treatment of neonatal HI encephalopathy.

The pathogenicity of Streptococcus parauberis isolated from cultured olive flounder Paralichthys olivaceus (양식 넙치, Paralichthys olivaceus에서 분리한 연쇄구균증 원인균, Streptococcus parauberis의 병원성)

  • Choi, Hee-Jung;Cho, Mi-Young;Lee, Jae-Il;Kwon, Mun-Gyeong;Choi, Dong-Lim;Kim, Jin-Woo;Han, Myoung-Chul;Lee, Deok-Chan
    • Journal of fish pathology
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    • v.22 no.3
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    • pp.263-273
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    • 2009
  • There is an increasing number of reports describing Streptococcus parauberis as an important pathogen of cultured olive flounder Paralichthys olivaceus and starry flounder Platichthys stellatus in Korea. We tried to determine the effects of water temperature (14${^{\circ}C}$ and 21${^{\circ}C}$) on the pathogenicity in Streptococcal disease caused by S. parauberis. We have challenged 180 olive flounder by i.p injection to $2.0{\times}10^{7}$ live cells/fish. Mortality was monitored for 21 days post challenge. And histopathological characterizations as infection degree, tissue degeneration and/or bacterial distribution were investigated with H&E stain and in situ hybridization technique. Fifty percent and 16.7% of mortality occurred within 21 days at 21${^{\circ}C}$ and 14${^{\circ}C}$ water temperature, respectively. In most cases, the typical symptoms of olive flounder infected with S. parauberis were darkness of the skin, lethargy, mild abdominal distension cause by ascites, splenomegaly, congested liver and internal organs paleness. The pericardial sac contained large amounts of cloudy fluid. Numerous whitish nodules, which were variable in size and often confluent, were randomly scattered throughout the myocardium. Especially, pericarditis and/or myocarditis was observed in all tested fishes after death. Positive in reaction with S. parauberis were found in all tissues in situ hybridization analysis. The relative numbers of S. parauberis in heart were much more than in liver, spleen, kidney and stomach. We evaluated that S. parauberis strain causes serious damage in the pericardium, shortness of breath and the blood disorder. Therefore, pericarditis and myocarditis caused by S. parauberis were closely related to mortality of olive flounder.

A STUDY ABOUT THE APOPTOSIS OF EPITHELIAL CELLS IN THE FUSING FETAL RAT PALATE (흰쥐 태아 구개융합부위 상피세포의 아포프토시스에 관한 연구)

  • Lee, Hong-Joo;Cha, Kyung-Suk;Lee, Jin-Woo
    • The korean journal of orthodontics
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    • v.31 no.1 s.84
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    • pp.71-83
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    • 2001
  • The purpose of this study was to Prove that the medial edge epithelial cells covering the secondary palatal shelves were removed by apoptosis during palatal fusion. 12 mature female rats (Suprague-Dawley) were mated overnight with male rats and sacrificed on days 15.0, 16.5, 16.75, 17.0 of pregnancy. The embryos were removed from the uterus and the heads were embedded in paraffin. The paraffin blocks were sectioned and the sections were undergone H-E staining for general histologic feature and TdT staining for detection of apoptotic cells. The obtained results were as follows. k. In the section of 16.0 and 16.5 day embryos, the palatal shelves were prior to contact and no apoptotic cells wereobserved in the medial edge epithelium. At the initial contact of Palatal shelves, there was a few apoptotic cells in the fusing epithelium. 2. In the 16.75 day embryos, the samples that epithelial seams did not lost there continuity, apoptotic cells were rarely seen at the midline epithelial seam. In contrast, a lot of apoptotic cells were observed at epithelial triangles and the junction between palatal shelves and nasal septum. 3. In the 16,75 day embryos, the samples that epithelial seams lost their continuity and disrupted to epithelial islands, large number, of apoptotic cells were observed at epithelial islands and epithelial triangles. Some apoptotic cells were also observed at the oral, nasal epithelium near the midline. 4. In the 17.0 day embryos, most of epithelial islands were disappeared and mesenchymal confluence was achieved. Apoptotic cells were rarely observed in the mesenchymal tissue which replaced epithelial islands, but there were some apoptotic cells at the epithelial triangles, oral and nasal epithelium. From the results of the study, it was revealed that medial edge epithelial cells of fusing palate were removed by apoptosis. Apoptotic cells were found mainly in the disappearing midline epithelial seam and the oral and nasal epithelial triangles at some late stages of palatal fusion.

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