• Title/Summary/Keyword: in vivo experiment

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Effect of Poly-L-arginine on the Mucin Release from Airway Goblet cells of Hamster and on the Mucosubstances of Airway Goblet cells of Rat (폴리-엘-아르기닌이 햄스터 기도 배상세포에서의 뮤신 유리 및 흰쥐 기도 배상세포내 함유된 점액에 미치는 영향)

  • 이충재
    • Biomolecules & Therapeutics
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    • v.9 no.4
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    • pp.263-269
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    • 2001
  • In this study, we tried to investigate whether poly-L-arginine (PLA) (MW 10,800) significantly affect mucin release from cultured hamster airway goblet cells and the mucosubstances of hypersecretory air-way goblet cells of rats. Confluent primary hamster tracheal surface epithelial (HTSE) cells were metabolically radiolabeled with $^3$H-glucosamine for 24 hr and chased for 30 min in the presence of varying concentrations of PLA to assess the effects on $^3$H-mucin release. Possible cytotoxicities of PLA were assessed by measuring both Lactate Dehydrogenate (LDH) release and by checking the possible changes on the morphology of HTSE cells during treatment. For in vivo experiment, hyperplasia of rat airway goblet cells and increase in intraepithelial mucosubstances were induced by exposing rats to SO$_2$ for 3 weeks and varying concentrations of PLA were administered inhalationally to assess the effects on the mucosubstances of airway goblet cells of rats. The results were as follows : (1) PLA significantly inhibited mucin release from cultured HTSE cells in a dose-dependent manner; (2) there was no significant release of LDH and no significant change on the morphology of cultured HTSE cells during treatment; (3) PLA also affected the intraepithelial mucosubstances of hypersecretory rats and restored them to the levels of control animals. We conclude that PLA inhibit mucin release from airway goblet cells without significant cytotoxicity and possibly normalize the hypersecretion of airway mucosubstances in vivo. This finding suggests that PLA might function as an airway mucoregulative agent.

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Kimchi cabbage (Brassica rapa L.) by-products treated with calcium oxide and alkaline hydrogen peroxide as feed ingredient for Holstein steers

  • Son, A-Rang;Kim, Seon-Ho;Valencia, Raniel A.;Jeong, Chang-Dae;Islam, Mahfuzul;Yang, Chul-Ju;Lee, Sang-Suk
    • Journal of Animal Science and Technology
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    • v.63 no.4
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    • pp.841-853
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    • 2021
  • This study aimed to investigate the effects of Kimchi cabbage by-products either treated or untreated with calcium oxide (CaO) and alkaline hydrogen peroxide (AHP) as substitutional ingredient of total mixed ration (TMR) on in vitro fermentation, in situ disappearance and growth performance of Holstein steers. Cannulated Holstein (600 ± 47 kg) was used for both the in vitro and in situ experiments. The treatments used were TMR only (CON), TMR + 30% Kimchi cabbage by-products fresh matter (FM) basis (TC), TMR + 30% Kimchi cabbage by-products FM basis + 5% CaO FM basis (TCC), and TMR + 30% Kimchi cabbage by-products FM basis + 5% CaO FM basis + 3.22% AHP FM basis (TCCA). For in vivo experiment, thirty-four Holstein steers (273 ± 45 kg) were subjected to a 150-day feeding trial, divided into two groups: CON and TC. In the in vitro experiment, pH of TCCA was greatest (p < 0.05) among other treatments at all incubation times. Ammonia nitrogen and volatile fatty acid concentrations were not significantly different for each treatment. However, butyrate was greater (p < 0.05) in TCC and CON than in both TC and TCCA. During in situ experiment, the dry matter (DM) disappearance was greatest (p < 0.05) in TCCA among other treatments. Also, disappearance of neutral detergent fiber (NDF) and acid detergent fiber (ADF) were observed greatest (p > 0.05) in TCCA treatment. In the in vivo experiment, average daily gain (ADG) did not differ between CON and TC. In blood profile analysis, alanine aminotransferase, aspartate aminotransferase, glucose, total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, and total protein concentration were not significantly different between treatments. But, creatinine concentration was greater (p < 0.05) in TC than in CON. Overall results suggest that Kimchi cabbage by-products either treated or untreated with CaO and AHP can be used as substitutional ingredient in TMR for Holstein steers.

Quantitative analysis of mutans streptococci adhesion to various orthodontic bracket materials in vivo (다양한 교정용 브라켓 원재료에 부착하는 mutans streptococci 양의 비교분석)

  • Yu, Jin-Kyoung;Ahn, Sug-Joon;Lee, Shin-Jae;Chang, Young-Il
    • The korean journal of orthodontics
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    • v.39 no.2
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    • pp.105-111
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    • 2009
  • Objective: To estimate the effects of bracket material type on enamel decalcification during orthodontic treatment, this study analyzed the adhesion level of mutans streptococci (MS) to orthodontic bracket materials in vivo. Methods: Three different types of orthodontic bracket materials were used: stainless steel, monocrystalline sapphire, and polycrystalline alumina. A balanced complete block design was used to exclude the effect of positional variation of bracket materials in the oral cavity. Three types of plastic individual trays were made and one subject placed the tray in the mouth for 12 hours. Then, the attached bacteria were isolated and incubated on a mitis salivarius media containing bacitracin for 48 hours. Finally, the number of colony forming units of MS was counted. The experiments were independently performed 5 times with each of the 3 trays, resulting in a total of 15 times. Mixed model ANOVA was used to compare the adhesion amount of MS. Results: There was no difference in colony forming units among the bracket materials irrespective of jaw and tooth position. Conclusions: This study suggested that the result of quantitative analysis of MS adhesion to various orthodontic bracket materials in vivo may differ from that of the condition in vitro.

SYNERGISTIC APOPTOTIC EFFECT OF TAXOL ON ORAL SQUAMOUS CELL CARCINOMA BY CYCLOSPORIN A (구강 편평세포암종에서 Taxol과 Cyclosporin A의 세포사멸 상승 작용 효과)

  • Suh, Min-Jung;Han, Se-Jin;Lee, Jae-Hoon
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.29 no.5
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    • pp.394-404
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    • 2007
  • Oral squamous cell carcinoma is the most prevalent oral cancer, which is characterized by its high metastasis and recurrent rates and poor prognosis. Taxol is an anticancer agent which is microbial products extracted from jew tree. It combines with the tubulin and induces apoptosis by inhibiting mitosis of cell with microtubule stabilization. Recently, it was reported to be effective in various solid tumors, but only very slight effect has been seen in oral squamous cell carcinomas due to its cell-specific potencies. Cyclosporin A is used as immune suppressant and is being applied in anticancer therapy as its mechanism of induction of change of apoptotic process in various cells have been known. In this study, oral squamous cell carcinoma HN22 cell line was used for in vitro experiment and as for the experimental group taxol and cyclosporin A were applied alone and to observe the synergistic effect of apoptosis, Taxol and cyclosporin A were coadministered with different concentration of taxol for comparison. The results were obtained as follow: 1. There was no difference in Bcl-2, Bax, caspase 3, 8, 9 mRNA expression when cyclosprin A or taxol was applied alone to HN 22 cell line. 2. Caspase 3, 9 mRNA expression was prominently increased when cyclosprin A and taxol were applied together to cancer cell. 3. No significant difference was observed when cyclosporin A and taxol($1{\mu}g/ml$ and $3{\mu}g/ml$) were applied together to cancer cell line. 4. No significant difference was seen in Bcl-2, Bax, and caspase 8 mRNA expression in all the groups of in vitro experiments. 5. When cyclosporin A was applied alone in vivo study on the nude mice, histopathologi cal findings was similar to those of the control group. Oral squamous cell carcinoma induced by inoculation of HN 22 cell line was not reduced after treatment of cyclosporin A. 6. When taxol was applied alone, the islands of squamous cell carcinoma still remained, which meant insignificant healing effect. There was a lesser volume increase compared with the cyclosporin A alone. 7. When taxol and cyclosporin A were applied together, the connective tissue and calcification were seen in the histopathologic findings. Oral squamous cell carcinoma was decreased and cancer cell was disappeared. In observing the tumor mass change with time, there was a gradual decreased size and healing features. As the results of the in vitro experiment, it could conclud that only when the two agents are applied together, mitochondria-mediated apoptosis occurred by considerable increase of caspase 3, 9 mRNA expression, irrespectable of the concentration of taxol. In vivo experiment, there was a discrete synergistic effect when the two agents were applied together. But single use of cyclosporin A was not effective in this study. Based on the results of this experiment, if further clinical studies are done, taxol and cyclosporin A could be effectively used in treatment of oral squamous cell carcinomas.

The Screening Condition for the Immune Regulatory Responsor Using Mouse Fetal Thymic Organ Culture (쥐의 태아 흉선 조직 배양을 이용한 면역조절제 검색방법 확립)

  • Lee, Seung-Gak;Song, Min-Dong;Lee, Kwang-Ho
    • Korean Journal of Pharmacognosy
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    • v.28 no.4
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    • pp.286-292
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    • 1997
  • We studied the screening condition for immune regulatory responsor. We focused on the T-lymphocytes leer this purpose. Mouse fetal thymic organ culture (FTOC) system and flow cytometric analysis were mainly used in this experiment. Even if FTOC is carried out in vivo condition, the pattern of thymic development in the condition of FTOC is similar to that of in vivo condition. In this regard, FTOC system might be very powerful tool to screen the immune regulator, especially concerning on T cells. To establish the optimum condition of FTOC to screen the Immune regulator, we focused on the optimum amount of dose and culture period. The cell number and surface antigens on T cells were also analysed by using hemacytometer and flow cytometer. To monitor the differentiation event, anti-CD3, anti-CD4 and anti-CD8 antibodies were used. Alkoxyglycerol and Phellodendri Cortex were used fur positive and negative control, respectively. Astragalus membranceus was used as test sample. From our analysis, we reached to conclusions that the best dose of extract is $50\;{\mu}g/ml$ of culture medium, the best culture period is for 9 days, and ethanol used as solvent has no toxicity to FTOC.

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Immune Enhancing Activity of Neoagarooligosaccharides from Marine Red Algae (홍조류 유래 네오아가로올리고당의 면역 활성 증강 효과)

  • Kim, Kyoung-Woon;Won, Ji-yeon;Kim, Eun Joo;Lee, Je-Hyeon;Lee, Miyeon;Na, Deukchae
    • The Korean Journal of Food And Nutrition
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    • v.35 no.1
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    • pp.43-50
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    • 2022
  • Agar, a heterogeneous polymer of galactose, is the main component of the cell wall of marine red algae. It is well established as a safe, non-digestible carbohydrate in oriental countries. Neoagarooligosaccharides (NAOs) prepared by hydrolyzing agar by microbial β-agarase have been reported to show safety. However, their immunological effects have not been reported yet. Thus, the objective of this study was to investigate immune enhancing effects of neoagarooligosaccharides (NAOs) from marine red algae Gelidium elegans in mice by performing ex vivo experiments. Six-week-old mice were fed ad libitum. NAOs were orally administrated at three different concentrations (100, 500, and 2,500 mg/kg B.W./day) twice a week for four weeks. The group fed with NAOs at 2,500 mg/kg showed the highest proliferation of splenocytes and production levels of cytokines (IL-1β, IL-6, TNF-α) in the ex vivo experiment. In conclusion, NAOs can enhance immune function, increase proliferation of splenocytes, and increase cytokine production by activating macrophages in mice.

Effect of Demineralized Bone Particle Gel Penetrated into Poly(lactic-co-glycolic acid) Scaffold on the Regeneration of Chondrocyte: In Vivo Experiment (PLGA 다공성 지지체에 함침시킨 DBP젤의 연골재생 효과: In Vivo 실험)

  • Lee, Yun Mi;Shim, Cho Rok;Lee, Yujung;Kim, Ha Neul;Jo, Sun A;Song, Jeong Eun;Lee, Dongwon;Khang, Gilson
    • Polymer(Korea)
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    • v.36 no.6
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    • pp.789-794
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    • 2012
  • Poly(lactic-co-glycolic acid) (PLGA) has been most widely used due to its advantages such as good biodegradability, controllable rate of degradation and metabolizable degradation products. We manufactured composite scaffolds of PLGA scaffold penetrated DBP gel (PLGA/DBP gel) by a simple method, solvent casting/salt leaching prep of PLGA scaffolds and subsequent soaking in DBP gel. Chondrocytes were seeded on the PLGA/DBP gel. The mechanical strength of scaffold, histology (H&E, Safranin-O, Alcian-blue) and immunohistochemistry (collagen type I, collagen type II) were performed to elucidate in vitro and in vivo cartilage-specific extracellular matrices. It was better to keep the characteristic of chondrocytes in the PLGA/DBP gel scaffolds than that PLGA scaffolds. This study suggests that PLGA/DBP gel scaffold may serve as a potential cell delivery vehicle and a structural basis for in vivo tissue engineered cartilage.

Design of a Magnetic Field Source for In Vivo Experiments at Extremely Low Frequency (생체 실험용 극저주파 자기장 발생 장치의 설계)

  • 김정호;김윤명
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
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    • v.14 no.8
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    • pp.871-877
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    • 2003
  • In this paper, the design parameters for the magnetic field source at extremely low frequency are proposed. This facility can be used fur in vivo experiments with small animals to investigate biological response to the driving magnetic fields. In case that the exposed animals are motionless, the animals may be affected by the directivity of driving field. To avoid this effect, a 2-axis ELF magnetic field driving apparatus was designed. The optimum location and number of turns of each coil were obtained by numerical analysis. Applying these data to the MATLAB code(for computation), the magnetic field distribution was obtained. The calculation result fur a well-designed facility showed that the space in which the amplitude of the magnetic field lies within the 95 % of the magnetic field distribution was more than 60 % of each axis length.

Effect of Polysaccharide from Schizophyllum commune on Burn and Wound Healing (치마버섯 유래 다당체의 화상 및 상처 치유효과)

  • Kim, Min Kyoung;Hong, Eock Kee
    • Korean Chemical Engineering Research
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    • v.44 no.1
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    • pp.87-91
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    • 2006
  • Polysaccharides extracted from mushroom have been most commonly used to enhance the immune system. Also polysaccharides maintaining the moisture extent on epidermal tissue have an effect on the removal of necrotic tissue and the restoration of epidermal tissue through enhancing the immune system at skin layers. In this work, polysaccharides were from Schizophyllum commune studied about the burn and wound healing activity in the epidermal tissue on rats through in vivo experiment and hematological values. And antibacterial activities were examined using pathogenic microorganisms causing the secondary inflammation.

The Effects of Gyejibokryeong-hwan (桂枝茯苓丸) for Wound Healing after Skin Suture (계지복령환(桂枝茯苓丸)이 피부봉합 후 상처 치유에 미치는 영향)

  • Jung, Hoon;Lee, Eun-Jung;Oh, Min-Seok
    • Journal of Korean Medicine Rehabilitation
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    • v.26 no.2
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    • pp.29-50
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    • 2016
  • Objectives There is few Korean medicinal studies about post-operation wound healing despite much effort for minimizing wound or post-op scar. The aim of this study is to evaluate the wound healing effect of Gyejibokryeong-hwan (Guizhifuling-wan, GBH) after skin suture. Methods < In vitro > We observed anti-oxidative and anti-inflammatory effect by using lipopolysaccharide (LPS)-treated RAW 264.7 cells. For anti-oxidation, we mesured the total amount of polyphenol, flavonoid, DPPH scavenging ability, ABTS scavenging ability and the value of ROS production, and for anti-inflammation, we mesured the amount of NO and pro-inflammatory cytokines ($TNF-{\alpha}$, $IL-1{\beta}$, IL-6). < In vivo > Thirty SD rats were divided into five equal groups (n=6, one normal, two controls and two experimentals). All groups except normal group were made a scar (around $1{\times}4cm^2$) in the back by the depth of the fascia and then sutured by a thread and needle. Normal group rats received no treatment at all. Control group rats were fed distilled water, and positive control group rats were percutaneously applied terramycin once in 2 days. GBH 200 group rats were orally medicated GBH 200 mg/kg, and GBH 400 group rats were orally medicated GBH 400 mg/kg per day for two weeks. We analyzed the blood samples (WBC, neutrophil, lymphocyte, monocyte, eosinophil), and the serums (TIMP-1, MMP-2, MMP-2. $PGE_2$, $TGF-{\beta}$, VEGF), and examined the wounded skin tissue histopathologically. Results < in vitro > 1. DPPH and ABTS scavenging activity was increased concentration-dependantly, and ROS production was significantly increased in GBH treated cells ($100{\mu}g/ml$). Therefore in this study, Gyejibokryeong-hwan appears to have the anti-oxidative. 2. NO production was significantly reduced in GBH treated cells ($100{\mu}g/ml$), and $IL-1{\beta}$ production was significantly reduced in GBH treated cells ($1{\mu}g/ml$). But, $TNF-{\alpha}$ and IL-6 did not show uneffective action. Therefore in this study, Gyejibokryeong-hwan did not show any significant effect on anti-inflammatory process. < in vivo > 1. Monocyte and neutrophil was significantly increased in GBH (200, 400) groups. WBC, lymphocyte and eosinophil did not show significant change. 2. TIMP-1, MMP-2, VEGF were significantly increased in GBH 400 group, $PGE_2$ was significantly reduced in GBH 400 group. $TGF-{\beta}$ was significantly increased in GBH (200, 400) groups, and MMP-9 was increased concentration-dependantly in GBH groups, but there was no significance. 3. In histopathological examinations, collagen was significantly increased and keratin was significantly decreased in GBH (200, 400) groups. Conclusions According to in vitro experiment, GBH appears to have the anti-oxidative effect and in vivo experiment, GBH stimulate the wound healing process hematologically and histopathologically. In conclusion, the results suggest that GBH promotes wound healing after skin suture.