• Title/Summary/Keyword: Histological analysis

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Anti-inflammatory effects of Ishige sinicola ethanol extract in LPS-induced RAW 264.7 cell and mouse model (LPS로 유도된 RAW 264.7 Cell과 마우스 모델에 대한 넓패(Ishige sinicola) 에탄올 추출물의 항염증 효과)

  • Kim, Ji-Hye;Kim, Min-Ji;Kim, Koth-Bong-Woo-Ri;Park, Sun-Hee;Cho, Kwang-Su;Kim, Go-Eun;XU, Xiaotong;Lee, Da-Hye;Park, Ga-Ryeong;Ahn, Dong-Hyun
    • Food Science and Preservation
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    • v.24 no.8
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    • pp.1149-1157
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    • 2017
  • Inflammation is the first response of the immune system to infection or irritation in our body. The use of medicinal plants has been widely applied as an alternative source for drug development. One of marine natural resources, the anti-inflammatory effect of Ishige sinicola ethanol extract (ISEE), was evaluated by using LPS-induced RAW 264.7 cell and mice model. As a result, the production of nitric oxide (NO) and pro-inflammatory cytokines (IL-6, IL-$1{\beta}$, TNF-${\alpha}$) were inhibited with increasing concentration of ISEE without any cytotoxicity. Furthermore, ISEE suppressed the expression of not only inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), nuclear factor-kappa B (NF-${\kappa}B$) p65, and mitogen-activated protein kinases (MAPKs), including extracellular signal-regulated kinase (ERK) 1/2, p38, and c-Jun N-terminal kinase (JNK) in a dose-dependent manner. In mice ear edema test, the formation of edema was reduced at the highest dosage of ISEE and the reduction of the number of infiltrated mast cells was observed in histological analysis. These results indicate that ISEE has a potent anti-inflammatory activity and can be used as a pharmaceutical material for many kinds of inflammatory disease.

Ser326Cys Polymorphism of hOGG1 Gene and Risk of Primary Lung Cancer in Koreans (한국인에서 hOGG1 유전자의 Ser326Cys 다형성과 원발성 폐암의 위험도)

  • Chae, Sang-Chul;Kim, Kyung-Rock;Joo, So-Young;Lee, Su-Yean;Kang, Kyung-Hee;Jeon, Kyung-Neoyh;Cha, Seung-Ick;Kim, Chang-Ho;Jung, Tae-Hoon;Park, Jae-Yang
    • Tuberculosis and Respiratory Diseases
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    • v.52 no.1
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    • pp.5-13
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    • 2002
  • Background: DNA repair plays a crucial role in protecting the genome from cancer-causing agents. Therefore, a reduced DNA repair capacity can increase the susceptibility to cancer. The human OGG1 (hOGG1) gene encodes DNA glycosylase/apurinic lyase and excise 8-hydroxyguanine, one of the major premutagenic DNA lesions, which is produced by oxygen radical forming agents including smoking. Recently several polymorphisms in the hOGG1 gene were identified, and it is possible that these polymorphism') may affect the DNA repair capacity and thus modulate cancer susceptibility. The relationship between the codon 326 polymorphism (Ser to Cys) in the hOGG1 gene and lung cancer risk was investigated. Materials and Method: The Ser326Cys genotypes were determined using PCR-RFLP analysis in 299 primary lung cancer patients and 186 healthy controls who were frequency (case:control=3:2) matched according to age and sex. Result: The frequencies of the Ser326Cys genotypes (Ser/Ser, Ser/Cys and Cys/Cys) among cases (23.4%, 51.8%, and 24.7%, respectively) were not significantly different from those among the controls (22.6%,52.1% and 25.3%, respectively). When the analyses were stratified according to age, sex, smoking status and packyears of smoking, no significant association between this polymorphism and lung cancer risk was found. Moreover, the Ser326Cys genotype showed no apparent relationship with any of the histological types of lung cancer. Conclusion: These result suggest that the hOGG1 Ser326Cys polymorphism is not a major contributor to individual lung cancer susceptibility in Koreans.

Collection, Identification and Hepatic Effect of Native Cordyceps militaris (새로운 번데기 동충하초의 수집, 동정 및 간기능에 미치는 효과)

  • Lee, Ki-Won;Nam, Byung-Hyouk;Jo, Wool-Soon;Oh, Su-Jung;Kang, Eun-Young;Cui, Yong;Lee, Jae-Yun;Cheon, Sang-Cheol;Jeong, Min-Ho;Lee, Jae-Dong
    • The Korean Journal of Mycology
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    • v.34 no.1
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    • pp.7-14
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    • 2006
  • Entomopathogenic fungus Cordyceps militaris is famous for its medicinal efficacies. It has been reported to have various pharmacological activities such as anti-tumour, insecticidal, antibacterial, immunomodulatory and antioxidant. In this study, we investigated the effect of the extract of C. militaris (MPUN8501), which was identified by the analysis of the nucleotide sequences of 5.8S ribosomal RNA, on the function of liver. C. militaris powder was extracted using hot water extracts method as time, volume and temperature and using method as differential polarity of organic solvent. Each fraction was tested for the improvement of hepatic enzyme alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activity. The BuOH extracts (CME) had highest activity which was used for the test of toxicity and efficacy of C. militaris. The enhancing effect of CME on the activity of ADH and ALDH was much more than medicine, drink, natural tea etc. Thus CME promoted the resolution of alcohol and acetaldehyde in rats, inducing recovery to normal condition rapidly. Furthermore, oral administration of CME effectively protected the carbon tetrachloride-induced acute hepatic injury as revealed by the hematological parameters (levels of sGOT and sGPT) and histological observation. CME was ascertained to be safe by regulatory toxicity studies of single dose toxicity and genotoxicity. These results suggest that CME would be useful for the maintaining normal hepatic activity as a functional health food.

Therapeutic Effects of Curdrania tricuspidata Leaf Extract on Osteoarthritis (골관절염 실험모델에서 꾸지뽕나무 추출물의 골관절염 억제효과 연구)

  • Nam, Da-Eun;Kim, Ok Kyung;Lee, Jeongmin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.5
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    • pp.697-704
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    • 2013
  • The inhibitory effect of ethanol extracts from Curdrania tricuspidata leaves (CTL) on osteoarthritis was investigated in primary cultured rat cartilage cells and a monosodium-iodoacetate (MIA)-induced arthritis rat model. To identify the effects of CTL 80% ethanol extracts (CTL80) and CTL 10% ethanol extracts (CTL10) against $H_2O_2$ treatment in vitro, cell survival was measured by the MTT assay. Cell survival after $H_2O_2$ treatment increased with CTL80 and CTL10 close to normal up to $300{\mu}g/mL\;H_2O_2$. The mRNA expression of matrix metalloproteinases (MMPs) was determined MMP-7 and MMP-13 (known catabolic factors), were significantly inhibited by CTL 80 and CTL10; a $200{\mu}g/mL$ dose of CTL80 especially decreased MMP-13 expression. In vivo, osteoarthritis was induced by an intra-articular injection of MIA into the knee joints of rats, then CTL80 and CTL10 orally administered daily for 35 days. After the animals were sacrificed, histological evaluations of their knee joints revealed a reduction in polymorphonuclear cell infiltration and smooth synovial lining in the CTL80-500 group. Micro-CT analysis of hind paws from CTL80-500 and CTL10 showed a protection against osteophyte formation, soft tissue swelling, and bone resorption. In conclusion, CTL ethanol extracts are effective in ameliorating joint destruction and cartilage erosion in MIA-induced rats. CTL decreases and normalizes articular cartilage through preventing extracellular matrix degradation and chondrocyte injury, and could potentially serve as a therapeutic treatment for humans.

Histological Analysis of Autologous Pericardial Tissue Used as a Small-Diameter Arterial Graft (소구경 동맥이식편으로 사용한 자가심란의 조직학적 분식)

  • Yang Ji-Hyuk;Sung Sang-Hyun;Kim Won-Gon
    • Journal of Chest Surgery
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    • v.39 no.4 s.261
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    • pp.261-268
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    • 2006
  • Background: Current vascular prostheses are still inadequate for reconstruction of small-diameter vessels. Autologous pericardium can be a good alternative for this purpose as it already possesses good blood compatibility and shows a mechanical behavior similar to that of natural arteries. However, the clinical use of autologous pericardial tissue as a small-diameter vascular graft has limitations due to mixed outcomes from uncertain biological behavior and difficulty to gain reliable patency results in animal experiments. To study this issue, we implanted fresh and glutaraldehyde-treated autologous pericardium as small-diameter arterial grafts in dogs, and compared their time-related changes histologically. Material and Method: As a form of 5mm-diameter arterial graft, one pair of autologous pericardial tissue was used for comparison between the glutaraldehyde-treated and the glutaraldehyde-untreated grafts in the bilateral carotid arteries in the same dog. The patency of the grafts were evaluated at regular intervals with Doppler ultrasonography. After the predetermined periods of 3 days, 2 weeks, 1 month, 3 months and 6 months, the grafts in each animal were explanted. The retrieved grafts were processed for light and electron microscopic analyses following gross observation. Result: Of 7 animals, 2 were excluded from the study because one died postoperatively due to bleeding and the other was documented as one side of the grafts being obstructed. All 10 grafts in the remaining 5 dogs were patent. Grossly, a variable degree of thromboses were observed in the luminal surfaces of the grafts at 3 days and 2 weeks, despite good patency. Pseudointimal smooth blood-contacting surfaces were developed in the grafts at f month and later. By light microscopy, mesothelial cell layers of the pericardial tissue were absent in all explanted grafts. Newly formed endothelial cell layers on the blood-contacting surface were observed in both the glutaraldehyde-treated and fresh grafts at 3 months and later. The collagen fibers became degraded by fragmentation in the fresh graft at 1 month and In the glutaraldehyde-treated graft at 3 months. At 6 months, the collagen layers were no longer visible in either the glutaraldehyde-treated or fresh grafts. By electron microscopy, a greater amount of coarse fibrin fibers were observed in the fresh grafts than in the glutaraldehyde-treated grafts and, more compact and well-arrayed layers were observed in the glutaraldehyde-treated grafts than in the fresh grafts. Conclusion: The glutaraldehyde-treated small-diameter pericardial arterial grafts showed a better endothelialization of the blood-contacting surface and a slower fragmentation of the collagen layers than the fresh grafts, although it has yet to be proven whether these differences are so significant as to affect the patency results between the groups.

In Vitro Effect of 808-nm Diode Laser on Proliferation and Glycosaminoglycan Synthesis of Rabbit Articular Chondrocytes (토끼 관절 연골세포의 증식과 글리코스아미노글리칸 합성에 대한 808-nm 다이오드 레이저의 효능 평가)

  • Minar, Maruf;Hwang, Ya-won;Choi, Seok-hwa;Kim, Gonhyung
    • Journal of Veterinary Clinics
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    • v.32 no.4
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    • pp.295-300
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    • 2015
  • The aim of the study was to assess the in vitro effect of 808-nm InGaAs diode laser on rabbit articular chondrocyte proliferation and sulphated glycosaminoglycan (sGAG) synthesis in alginate bead. Previous studies revealed either positive or negative stimulatory effects of laser on different types of cells. A 808-nm InGaAs diode laser at 1.0W power output was used to irradiate the rabbit chondrocytes in alginate beads with energy densities of $31J/cm^2$ (G 1) and $62J/cm^2$ (G 2) corresponding to the experimental groups for 10 seconds and 20 seconds, respectively at 24, 48, 72 and 96 hours after seeding. Control group was left untreated. MTT assay was performed at 1 week and 2 weeks after the $1^{st}$ laser irradiation in alginate beads. sGAG synthesis in alginate beads at 1 week and 2 weeks were determined by DMMB assay. Histological evaluation for cellular distribution and sGAG deposition around the cells were performed by alcian blue stain. MTT assay revealed no positive stimulatory effect in cell proliferation in alginate bead. DMMB assay results showed significantly increased sGAG production in G 2 chondrocytes at 2 weeks. Image analysis of alcian blue stained slides also showed significantly higher percentage of positive alcian blue stain in G 2 chondrocytes. This result suggests that 808-nm InGaAs diode laser with 1.0 W power output although cannot stimulate cell proliferation it can increase the cell secretion activity and sGAG deposition in alginate beads.

AGL gene mutation and clinical features in Korean patients with glycogen storage disease type III

  • Ko, Jung-Min;Kim, Gu-Hwan;Yoo, Han-Wook
    • Journal of Genetic Medicine
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    • v.4 no.1
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    • pp.72-79
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    • 2007
  • Purpose : Glycogen storage disease type III (GSD-III) is a rare autosomal recessive disorder of glycogen metabolism. The affected enzyme, amylo-1,6-glucosidase, 4-alpha-glucanotransferase (AGL, glycogen debranching enzyme), is responsible for the debranching of the glycogen molecule during catabolism. The disease shows clinical and biochemical heterogeneity, reflecting genotype-phenotype heterogeneity among different patients. In this study, we aim at analyzing mutations of the AGL gene in three unrelated Korean GSD-III patients, and characterizing their clinical and laboratory findings. Methods : We characterized the clinical features of three unrelated Korean GSD-III patients by biochemical, histological and imaging studies. The 35 exons and part of exon-intron boundaries of AGL were analyzed by direct sequencing using genomic DNA extracted from the peripheral leukocytes of patients. Results : Diverse clinical features were observed in these patients including hepatomegaly (all patients), seizures (patient 2), grow th failure (patients 1 and 2), hyperlipidemia (patients 1 and 3), raised transaminase and creatine kinase concentrations (all patients), and mild cardiomyopathy (patient 2). Liver transplantation w as performed in patient 2 due to progressive hepatic fibrosis. A dministration of uncooked corn starch maintained normoglycemia and improved biochemical and growth profiles. DNA sequence analysis revealed mutations in 5 out of 6 alleles. Patient 1 was a compound heterozygote of c.1282 G>A (p.R428K) and c.1306delA (p.S603PfsX6), patient 2 had c.1510_1511insT (p.Y 504L fsX 10), and patient 3 had c.3416 T >C (p.L 1139P) and c.1735+1 G>T (p.Y 538_R578delfsX 4) mutations. A part from the p.R428K mutation, the 4 other substitutions identified w ere nov el. Conclusion : GSD-III patients display variable phenotypic characteristics resembling those of GSD-Ia. Molecular defects in the AGL gene of Korean GSD-III patients are genetically heterogeneous.

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EVALUATION OF PERIODONTAL LIGAMENT CELL VIABILITY IN RAT TEETH AFTER FROZEN PRESERVATION USING IN-VIVO MTT ASSAY (급속냉동된 쥐 치아의 in vivo MTT 검색법을 이용한 치주인대세포 활성도 평가)

  • Kim, Jae-Wook;Kim, Eui-Sung;Kim, Jin;Lee, Seung-Jong
    • Restorative Dentistry and Endodontics
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    • v.31 no.3
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    • pp.192-202
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    • 2006
  • The purpose of this study was to examine the viability of PDL cells in rat molars by using in vivo MTT assay, which was used to compare fast cryopreservation group by liquid nitrogen $(-196^{\circ}C)\;with\;4^{\circ}C$ cold preservation group. A total of 74 Sprague-Dawley white female rats of 4 week-old with a body weight of 100 grams were used. The maxillary left and right, first and second molars were extracted as atraumatically as possible under ketamine anesthesia. Ten teeth of each group were divided as six experimental groups depending upon the preservation. Cryopreservation groups were Group 1 (5% DMSO 6% HES in F medium) Group 2 (10% DMSO in F medium), Group 3 (5% DMSO 6% HES in $Viaspan^(R)$). Group 4 (10% DMSO in $Viaspan^(R)$) which were cryopreserved for 1 week and cold preservation groups were Group 5 (F medium) , Group 6 ($Viaspan^(R)$) at $4^{\circ}C$ for 1 week. Immediate extraction group was used as a control. After preservation and thawing, the in vivo MTT assay was processed. Two way ANOVA and Duncan's Multiple Range Test was performed at the 95 % level of confidence, Another 2 teeth of each group were treated as the same manner and frozen sections $10{\mu}m$ thick for microscopic observation. The value of optical density obtained after in vivo MTT analysis was divided by the value of eosin staining for tissue volume standardization. Group 1, 2 had significantly higher optical density than Group 3 and 4 which had the lowest OD value. Group 6 had higher OD value than in Group 5 (P<0.05). Histological findings of periodontal ligament cell, after being stained with MTT solution were consistent with the in vivo MTT assay results. In this study, the groups which were frozen with DMSO as a cryoprotectant and the groups with F medium showed the best results.

Effect of the Flavonoid Luteolin for Dextran Sodium Sulfate-induced Colitis in NF-${\kappa}B^{EGFP}$ Transgenic Mice (Dextran Sodium Sulfate 유발 장염 모델에서 루테올린의 치료효과)

  • Jang, Byung-Ik
    • Journal of Yeungnam Medical Science
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    • v.23 no.1
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    • pp.26-35
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    • 2006
  • Background: Luteolin, a flavone found in various Chinese herbal medicines is known to possess anti-inflammatory properties through its ability to inhibit various proinflammatory signaling pathways including NF-${\kappa}B$ and p38 MAPK. In this study, we investigated the potential therapeutic effect of luteolin on dextran sodium sulfate (DSS)-induced colitis. Materials and Methods: We used a transgenic mouse model expressing the enhanced green fluorescent protein (EGFP) under the transcriptional control of NF-${\kappa}B$ $cis$-elements. C57BL/6 NF-${\kappa}B^{EGFP}$ mice received 2.5% DSS in their drinking water for six days in combination with daily luteolin administration (1mg/kg body weight, 0.1ml vol, intragastric) or vehicle. NF-${\kappa}B$ activity was assessed macroscopically with a Charge-Coupled Device (CCD) camera and microscopically by confocal analysis. Results: A significant increase in the Disease Activity Index (DAI), histological score (p<0.05), IL-12 p40 secretion in colonic stripe culture (p<0.05) and EGFP expression was observed in luteolin and/or DSS-treated mice compared to water-treated mice. Interestingly, a trend toward a worse colitis (DAI, IL-12p40) was observed in luteolin-treated mice compared to non-treated DSS-exposed mice. In addition, EGFP expression (NF-${\kappa}B$ activity) strongly increased in the luteolin-treated mice compared to control mice. Confocal microscopy showed that EGFP positive cells were primarily lamina propria immune cells. Conclusions: These results suggest that luteolin is not a therapeutic alternative for intestinal inflammatory disorders derived for primary defects in barrier function. Thus, therapeutic intervention targeting these signaling pathways should be viewed with caution.

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Changes in Biochemical Components of Several Tissues in Solen grandis, in Relation to Gonad Developmental Phases (대맛조개, Solen grandis의 생식소 발달 단계에 따른 일부 조직의 생화학적 성분변화)

  • Chung, Ee-Yung;Kim, Hyun-Jin;Kim, Jong-Bae;Lee, Chang-Hoon
    • The Korean Journal of Malacology
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    • v.22 no.1 s.35
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    • pp.27-38
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    • 2006
  • We investigated the reproductive cycle with gonad developmental phases of Solen grandis by histological observations. Seasonal changes in biochemical components of the adductor muscle, visceral mass, foot muscle and mantle were studied by biochemical analysis, from January to December, 2005. The reproductive cycle of this species can be classified into five successive stages: early active stage (December to January), late active stage (January to March), ripe stage (March to July), partially spawned stage (June to July) and spent/inactive stage (July to December). Total protein content was the highest in the foot muscle, the content was high in January (early active stage), the lowest in April (ripe stage), and was the highest in August (partially spawned stage). In the visceral mass, total protein content began to increase in February (late active stage) and reached a maximum in March (ripe stage). Thereafter, it gradually decreased between June and July (partially spawned stage). There was a strong negative correlation in total protein contents between visceral mass and mantle (r = -0.594, p = 0.042). Meanwhile there was a positive correlation between the adductor muscle and foot muscle, the correlation was not statistically significant (r = 0.507, p = 0.093). Total lipid content was the highest in the visceral mass; it was more than 2 to 5-fold higher than that in the adductor muscle, foot muscle, and mantle. Monthly changes in total lipid content were also most dynamic in the visceral mass. It was relatively higher between January and February, showed a maximum in March (the ripe stage), decreased rapidly from April to July (ripe and partially spawned stage), and gradually decreased from September to December (spent/inactive stage). There was a strong positive correlation in total lipid content between foot muscle and adductor muscle (r = 0.639, p = 0.025). Tthough a negative correlation was found between visceral mass and mantle (r = -0.392), the correlation was not statistically significant (p = 0.208). Glycogen contents changed within relatively narrow range and were similar among different tissues. There was no statistically significant correlation in glycogen contents among tissues.

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