• Title/Summary/Keyword: H & E staining

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Effect of Hominis Placenta on cutaneous wound healing in normal and diabetic mice

  • Park, Ji-Yeun;Lee, Jiyoung;Jeong, Minsu;Min, Seorim;Kim, Song-Yi;Lee, Hyejung;Lim, Yunsook;Park, Hi-Joon
    • Nutrition Research and Practice
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    • v.8 no.4
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    • pp.404-409
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    • 2014
  • BACKGROUND/OBJECTIVES: The number of diabetic patients has recently shown a rapid increase, and delayed wound healing is a major clinical complication in diabetes. In this study, the wound healing effect of Hominis placenta (HP) treatment was investigated in normal and streptozotocin-induced diabetic mice. MATERIALS/METHODS: Four full thickness wounds were created using a 4 mm biopsy punch on the dorsum. HP was injected subcutaneously at the middle region of the upper and lower wounds. Wounds were digitally photographed and wound size was measured every other day until the 14th day. Wound closure rate was analyzed using CANVAS 7SE software. Wound tissues were collected on days 2, 6, and 14 after wounding for H/E, immunohistochemistry for FGF2, and Masson's trichrome staining for collagen study. RESULTS: Significantly faster wound closure rates were observed in the HP treated group than in normal and diabetes control mice on days 6 and 8. Treatment with HP resulted in reduced localization of inflammatory cells in wounded skin at day 6 in normal mice and at day 14 in diabetic mice (P < 0.01). Expression of fibroblast growth factor (FGF) 2 showed a significant increase in the HP treated group on day 14 in both normal (P < 0.01) and diabetic mice (P < 0.05). In addition, HP treated groups showed a thicker collagen layer than no treatment groups, which was remarkable on the last day, day 14, in both normal and diabetic mice. CONCLUSIONS: Taken together, HP treatment has a beneficial effect on acceleration of cutaneous wound healing via regulation of the entire wound healing process, including inflammation, proliferation, and remodeling.

Histopathologic Changes of Articular Cartilage and Subchondral Bone in Cylindrical Biopsy Specimen from Talar Osteochondral Lesions (거골의 골연골 병변의 원주형 생검에서 관절 연골과 연골하 골의 조직병리학적 변화)

  • Lee, Ho-Seong;Jang, Jae-Suk;Lee, Jong-Suk;Cho, Kyung-Ja;Lee, Sang-Hoon;Jung, Hong-Keun;Kim, Yong-Min
    • Journal of Korean Foot and Ankle Society
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    • v.10 no.2
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    • pp.117-124
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    • 2006
  • Purpose: This study was aimed at elucidating the pathogenesis of talar osteochondral lesion by analyzing the histopathological findings. Materials and Methods: Twenty specimens from 20 patients who underwent surgical treatment for talus osteochondral lesions were studied. Preoperative MRI images including T1, T2, and stir images were taken and cases were classified according to modification of the Anderson's classification. There were 5 cases of MRI group 1, 6 cases of group 2, 7 cases of group 3 and 2 cases of group 4. A full thickness osteochondral plug including the osteochondral lesion of the talus was harvested from each patient and reviewed histopathologic changes of osteochondral fragment using H-E staining. Mean diameter of specimens was 8.5 mm and mean depth was 10.3 mm. Pathologic changes of articular cartilage and subchondral bone were observed. Subchondral bone was divided into superficial, middle and deep zones according to depth. Cartilage formation, trabecular thickening and marrow fibrosis were observed in each zone. Results: There were detachment of the joint cartilage at the tidemark in 16 cases of 20 cases and the separated cartilages were almost necrotic on the histopathologic findings. Cartilage formation within subchondral bone was discovered beneath the tidemark in 12 cases. Trabeculae were increased and thickened in 17 cases. These pathologic changes were similar to fracture healing process and these findings were more conspicuous near the tidemark and showed transition to normal bone marrow tissue with depth. No correlation between the pathological progression and MRI stages was found. A large cyst shown on MRI's was microscopically turned out to be multiple micro-cysts accompanied by fibrovascular structure and newly formed cartilage tissue. Conclusion: The histopathologic findings of osteochondral lesions are detachment of overlying cartilage at the tidemark and subsequent changes of subchondral bone. Subchondral bone changes are summarized as cartilage formation, marrow fibrosis and trabecular thickening that mean healing process following repeated micro fractures of trabecular. These osteochondral lesions should have differed from osteochondral fractures.

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Id3 mRNA Expression on Folliculogenesis in Rat Ovary (쥐 난소에서 난포 발달에 따른 Id3 mRNA의 발현)

  • Hwang, Seong-Soo;Ko, Yeoung-Gyu;Lim, Hyun-Joo;Seong, Hwan-Hoo;Yoon, Jong-Taek;Min, Kwan-Sik
    • Reproductive and Developmental Biology
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    • v.31 no.3
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    • pp.181-186
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    • 2007
  • Inhibitor of DNA binding protein or inhibitor of differentiation(Id) is largely considered as positive and/or negative regulators of proliferation, differentiation, angiogeneisis, and apoptosis. The four Id genes(Id1, Id2, Id3, and Id4) were known in mammals. However, little is known about the expression and function of these genes in reproductive physiology. Among them, this study was conducted to analyze the expression pattern of Id3 mRNA on folliculogenesis in rat ovary. After PMSG administration, the ovaries were obtained at 3, 6, 12, 24, 36, and 48hrs, fixed, dehydrated, and paraffin embedded. For in situ hybridization, anti-sense and sense Id3 cRNA probes were prepared and applied to the ovarian section. The ovarian sections were coated with NTB-2 emulsion. After that, the slides were developed and counterstained with hematoxylin and eosin staining. The hybridization signal was estimated on a scale of 1+ to 4+. In oocyte, the intensity of Id3 mRNA in primordial and primary follicles was scored at ${\geq}2+$, but the intensity was less than 1+ in secondary, dominant, and preovulatory follicles. In granulosa cells, the Id3 mRNA was strongly expressed(3+ or 4+) in dominant and preovulatory follicles. Taken together, Id3 mRNA was expressed specifically at follicle stages and follicular tissue and might be closely related with follicle development.

A Study on the Ultrastructure of Reproductive Organ of Korean Planaria (Dugesia japonica) (한국산(韓國産) 플라나리아(Dugesia japonica Ichikawa et Kawakatsu)생식기관(生殖器官)의 미세구조(微細構造)에 관(關)한 연구(硏究))

  • Chang, N.S.;Kim, W.K.
    • Applied Microscopy
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    • v.15 no.1
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    • pp.31-58
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    • 1985
  • The morphological study on different types of cells of reproductive organ including spermatogenesis in the adult planaria was performed to observe their cytochemical and ultrastructural characteristics. 1. Spermatogenesis The circular luminated material appears immediately inside the nuclear envelope of early spermatid and is found also in the nucleus of sperm, but typical acrosomal structures cannot be observed. Approximately ten of small-sized mitochondria occur around the nucleus in the transitional phase from primary spermatocyte to secondary spermatocyte, but in sperm a long mitochondrion is closely associated with nucleus, parellel to long axis of it. The sperm has a relatively long head connected with two tails via hollow neck. 2. Reproductive organ The penis bulb and the bursa stalk were observed. (1) Penis bulb The cells constituted penis bulb are classified into six types on the basis of ultrastructure of the cells and cytochemistry of the cytoplasmic granules. 1) A-type cells: These cells exhibiting low electron density are mainly occupied by large nucleus. These cells possess two different types of granules: highly electron-dense round granules with an average size of $0.9{\mu}m$, and electron-dense granules exhibit PAS-positive reaction. 2) B-type cells contain PAS-positive granules with the size of about $0.4{\mu}m$. They are rich in free ribosomes and mitochondria. 3) C-type cells are found to be dark cells due to high electron-density. These cells are largely occupied by large nucleus. 4) D-type cells: These cells are seen as light cells which have poorly developed cell organelles. 5) E-type tells: These cells contain a large number of glycogen granules which occupy most of cell. 6) F-type cells: These arc parietal epidermal cells surrounding the genital antrum. These cells are characterized by their finger-like shapes and the presence of a number of electron-dense, irregularly-shaped structures inside cells. The relatively large electron-lucent granules can be also found. The F-type cells possess numerous microvilli on their free surfaces. (2) Bursa stalk The cells constituted bursa stalk are classified into 3 types on the basis of cell shapes and presences of electron-dense or electron-lucent granules. 7) G-type cells with a long cytoplasmic process. They have large nuclei and poorly developed cell organelles. 8) H-type cells: These cells are characterized by the presence of a long cytoplasmic process and relatively highly electron-dense cytoplasmic profile. They have poorly developed cell organelles. 9) I-type cells contain large electron-lucent granules which exhibit negative reactions with three kinds of cytochemical staining methods used in this experiment. The fine electron-dense structures can be found inside these granules.

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Expression of p53 and Ki-67 in Cervical Dysplasia with Human Papilloma Virus Infection or Non-infection (인유두종 바이러스의 감염 또는 감염되지 않은 자궁 경부 이형성증에서 p53 및 Ki-67의 발현)

  • Choi, Sook-Kyung;Kim, Tai Jeon;Hong, Seung Bok;Lee, Hun Taeck
    • Korean Journal of Clinical Laboratory Science
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    • v.36 no.2
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    • pp.178-184
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    • 2004
  • This research focuses on the overall evaluation of tumor protein (p53) and cell growth marker (Ki-67) in their functions as carcinogenic factors in both a HPV-infected group and in a HPV-noninfected group with the precancerous dysplasia of uterine cervix. Histological grades were determined by the H&E staining and the expression level of p53 and Ki-67 were tested by the immunohistochemistry method. The results were as follows. Among the total of 66 cases, p53 (+) was observed in 19 cases (29.0%) from the mild grade group, 22 cases (33.0%) from the moderate grade group, and 19 cases (29.0%) from the severe grade group. The values correlate with the increase of dysplasia intensity in HPV-noninfected group and showed significant correlation (p<0.05), but there were no significant difference from the HPV-infected group. Among a total of 66 cases, the mitotic index of Ki-67 (+) were observed in 19 cases (29.0%) from the mild grade group, 22 cases (33.0%) from the moderate grade group, and 19 cases (29.0%) from the severe grade group. The values were significantly different against dysplasia intensity (p<0.05), but showed no significant difference from HPV infection. After cross comparing the statistical parameters of p53 and ki-67 in their significance, p53 was shown to be statistically significant with Ki-67 while there was no statistically significant difference with Ki-67 (p<0.05). Taken together, tumor protein (p53) and an index of Ki-67 observed in cervical dysplasia and in HPV related dysplasia of cervix uterine did not have any notable significance with HPV infection. The incidence rate of p53, however, had some significant correlation with dysplasia while Ki-67 had no particular statistical significance. As a result, p53 and Ki-67 can be considered as effective diagnostic markers in predicting the disease progression of dysplasia to cervical cancer.

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Effect of Lactic Acid Bacteria Powder on Loperamide-induced Constipation in Rat (Loperamide로 유도된 변비 증상에 유산균 제제가 미치는 영향)

  • Kim, Eun Young;Jo, Kyungae;Ahn, So Hyun;Park, Sung Sun;Son, Heung Soo;Han, Sung Hee
    • The Korean Journal of Food And Nutrition
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    • v.28 no.6
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    • pp.956-964
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    • 2015
  • Probiotics is known improve the microenvironment of colon; however, the metagenomic DNA study of its lactic acid bacteria in constipation induced by loperamide is not clearly understood. In the present study, we investigated the reduction of the lactic acid bacteria in case of constipation, in normal and loperamide-induced rat. Lactic acid powder (lactic acid bacteria 19) was prepared from Chong Kun Dang Pharmaceutical Corporation. After 2 weeks of oral administration, the group treated with the higher concentration of lactic acid bacteria ($10^9CFU/mL$ per kg of body weight) following loperamide treatment was the most effective in increasing number, weight, and water content of feces. A similar but significant increase was found in the group treated with lower concentration of lactic acid bacteria ($10^7CFU/mL$ per kg of body weight) after loperamide treatment. The concentrations of acetic acid and propionic acid in feces in the loperamide-induced rat with high concentration lactic acid, were significantly higher than that of others. Furthermore, gastrointestinal transit ratio as well as the length and area of intestinal mucosa were significantly increased after treatment with lactic acid bacteria in loperamide-induced rat. Metagenomics DNA analysis indicated that the microorganism homology in cecum was similar between the groups of normal (NOR) and HIG. Our results show that lactic acid bacteria were effective in improving the constipation.

Anti-obesity and Anti-inflammation Effects of Cheonggukjang in C57Bl/6 mice with High Fat Diet Induced Obesity (고지방식이로 유도된 비만 마우스에서 청국장의 항비만 및 항염증 효과)

  • Kim, Jiyoung
    • Journal of Life Science
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    • v.27 no.11
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    • pp.1357-1368
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    • 2017
  • The purpose of this study was to investigate the anti-obesity and anti-inflammation effect of the cheonggukjang (a soybean paste fermented for only a few days) in diet induced obesity mice. Weight gain was significantly decreased in the mice fed cheonggukjang compared High Fat Diets (HFD). The HFD plus cheonggukjang (CGJ) were also effective in improving the lipid metabolism. The levels of plasma triglyceride, cholesterol, ALT, AST, leptin, glucose, and insulin were significantly lower in CGJ than HFD group (p<0.05). The adiponectin level of CGJ group was significantly increased compared to the HFD group (p<0.05). In the CGJ group, the mRNA expression of adipogenic genes in the liver and adipose tissues, which are transcription factors crucial for adipogenesis, were significantly suppressed (p<0.05). The number of $CD11b^+F4/80^+$ T cells, $Gr-1^{int}CD11b^{high}$ cells, and $Gr-1^{int}CD11b^{high}$ cells were significantly higher in HFD group than CGJ group (p<0.05). The size of adipocyte was significantly reduced in CGJ group compared to HFD group. In addition, the contents of liver lipid droplets were significantly downregulated in the CGJ mice than HFD mice (p<0.05). Collectively, these data suggest the novel function of cheonggukjang in modulating adipogenesis through an immune function-alteration involving downregulation of adipogenic transcription factors and macrophage activation.

Evaluation of Various Scaffolds for Tissue Engineered Biodisc Using Annulus Fibrosus Cells (조직공학적 바이오디스크의 섬유륜 재생을 위한 지지체 특성평가)

  • Ha, Hyun-Jung;Kim, Soon-Hee;Yoon, Sun-Jung;Park, Sang-Wook;So, Jung-Won;Kim, Moon-Suk;Rhee, John-M.;Khang, Gil-Son;Lee, Hai-Bang
    • Polymer(Korea)
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    • v.32 no.1
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    • pp.26-30
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    • 2008
  • This study was designed to investigate the effect of hybridization of synthetic/natural materials for annulus fibrosus (AF) tissue regeneration in vitro and in vivo. The synthetic/natural hybrid scaffolds were prepared using PLGA (poly (lactic-co-glycolic) acid), SIS (small intestinal submucosa) and DBP (demineralized bone particles). PLGA, PLGA/SIS(20%), PLGA/DBP(20%) and PLGA/SIS (10%)/DBP (10%) scaffold were manufactured by solvent casting/salt leaching method. Compressive strength was measured. Rabbit AF cells were isolated, cultured and seeded into experimental groups. Hydroxyproline production and DNA quantity of AP cells on each scaffold was measured at 2, 4 and 6 weeks after in vitro culture. Cell-scaffold composites were implanted subcutaneously into athymic mice. After 1,4 and 6 weeks postoperatively, specimens were taken and H&E, Safranin-O and type I collagen staining were carried out concerning formation of cartilagenous tissue. In vitro PLGA/SIS scaffold was evaluated for total collagen content (bydroryproline/DNA content) and PLGA scaffold was evaluated for compressive strength.

Comparison of Anti-asthmatic Activity by Native Codonopsis lanceolata Extract (자생 돌더덕 추출물에 의한 천식억제 활성의 분석)

  • Lee, Seung-Ha;Choi, Hee-Jeong;Heo, Jin-Chul;Lee, Jong-Ha;Kwon, Taeg Kyu;Ha, Sang-Chul;Lee, Sang-Han
    • Journal of Life Science
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    • v.27 no.4
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    • pp.450-455
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    • 2017
  • Codonopsis lanceolata (Campanulaceae) has been widely used in traditional medicine and is considered to have medicinal properties to treat diseases and symptoms such as bronchitis, coughs, spasm, edema, hepatitis, colitis, and lung injury. In order to investigate whether native Codonopsis lanceolata extract alleviates ashmatic symptoms in vivo, we first carried out various antioxidant activities by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, ferric reducing antioxidant power (FRAP), and cupric reducing antioxidant capacity (CUPRAC) assays. The antioxidant activities were increased by adding Codonopsis lanceolata extract in a concentration-dependent manner which compared to ascorbic acid as a positive control. Histological studies using an ovalbumin-induced animal model exhibited potent anti-inflammatory potential by decreasing immuno-responsive cells in the lung by the extract by confirming H&E and PAS staining. It is revelaed that further immunihistochemical analysis showed anti-ashmatic capabilities by assessing histamine, IL-31, and MMP-9 expressions. The level of IL-13 expression in Codonopsis lanceolata extract-treated group was decreased upto 73.7% compared to control, whereas that of total cells and eosinophil counting in Codonopsis lanceolata extract-treated group was diminished to 73.5% and 80.9%, respectively. These results collectively indicate that the C. lanceolata extract ameliorates asthmatic symptoms effectively in an ovalbumin-challenged mice model, in that the extract can be used for the development of an anti-asthmatic food ingredient.

Bone formation effect of $HA/{\beta}-TCP$ composite powders in rabbit calvarial bone defects;Histologic study (토끼의 두개골내에 형성된 골결손부에서 $HA/{\beta}-TCP$ composite powders의 골형성에 관한 조직학적 연구)

  • Lee, Kwang-Ho;Jang, Hyun-seon;Park, Joo-Cheol;Kim, Heung-Joong;Kim, Chong-Kwan;Kim, Byung-Ock
    • Journal of Periodontal and Implant Science
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    • v.36 no.1
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    • pp.1-14
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    • 2006
  • The purpose of the present study was to evaluate the histologic results of bone cavities that were surgically created in the calvaria of rabbit and filled with $HA/{\beta}-TCP$ composite powders, which had been developed in Korea (Dentium, Korea). Ten young adult rabbits were used. Four defects were surgically produced in calvaria of each rabbit. Each rabbit was anesthetized with Ketamine-HCI (5 mg/kg, Yuhan Cor. Korea) and Xylazine-HCI (1.5 ml/kg, Yuhan Cor. Korea)). An incision was made to the bony cranium and the periosteum was reflected. Using a trephine bur (external diameter: 8 mm, 3i, USA), 4 'through-and-through' bone defects were created with copious irrigation, and classified into 4 groups: control group: no graft materials, experimental group I: normal saline + graft materials: experimental group II: venous blood + graft materials: experimental group III: graft materials only. The defects were randomly filled with graft materials. The defects were closed with resorbable suture material. At the end of the surgical procedure, all animals received a single intramuscular injection of antibiotics Gentamicin (0.1 mg/kg, Dae Sung Microb. Korea). Rabbits were sacrificed with phentobarbital (100 mg/kg) intravenously at 1-, 2-, 4-, 6- and 8-week after. Specimens were treated with hydrochloric acid decalcifying solution (Fisher Scientific, Tustin, CA) and sectioned by bisecting the 8 mm diameter defects. The histologic specimens were prepared in the general method with H & E staining at 6 ${\mu}m$ in thickness. The results were as follows; 1. New bone formation showed from after 2-week of surgery in defect area. As time lapsed, lots of new bone formation and mature bones showed. 2. Histologically, degree of new bone formation could not be discerned among the experimental groups. But, for experimental group II, lots of cells gathered around graft materials after 1-week of surgery, new bone formed slightly faster and than the others at 1-week after. For experimental group I, a few inflammatory finding showed around graft material at after 1-week and after 2-week of surgery. 3. No bone formation did show for control group. Based on histologic results, the new $HA/{\beta}-TCP$ composite powders appeared to act as a scaffolding material for regeneration of osseous defects.