• Title/Summary/Keyword: In vivo

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Apoptotic Effect of co-treatment with HS-1200 and Cisplatin on SCC25 Human Tongue Squamous Cell Carcinoma Cell Line (HS-1200과 cisplatin의 병용처리가 사람구강암세포에 미치는 세포자멸사 효과에 대한 연구)

  • Kim, Duk-Han;Kim, In-Ryoung;Park, Bong-Soo;Ahn, Yong-Woo;Jeong, Sung-Hee
    • Journal of Oral Medicine and Pain
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    • v.38 no.3
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    • pp.221-233
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    • 2013
  • Bile acids are polar derivatives of cholesterol essential for the absorption of dietary lipids and regulate the transcription of genes that control cholesterol homeostasis. Recently it have been identified the synthetic chenodeoxycholic acid (CDCA) derivatives HS-1200 and cisplatin showed apoptisis-inducing activity on various cancer cells in vivo and in vitro. This study was undertaken to investigate the synergistic apoptotic effect of co-treatment with HS-1200 and cisplatin on human tongue squamous cell carcinoma cells (SCC25 cells). To investigate whether the co-treatment with HS-1200 and cisplatin compared to each single treatment efficiently reduces the viability of SCC25 cells, MTT assay was conducted. The induction and augmentation of apoptosis were confirmed by DNA electrophoresis, Hoechst staining and an analysis DNA hypoploidy. Westen blot analysis and immunofluorescent staining were also performed to evaluate the expression levels and the translocation of apoptosis-related proteins following this co-treatment. Furthermore, proteasome activity and mitochondrial membrane potential (MMP) change were also assayed. In this study, co-treatment with HS-1200 and cisplatin on SCC25 cells showed several lines of apoptotic manifestation such as nuclear condensations, DNA fragmentation, reduction of MMP and proteasome activity, the increase of Bax and the decrease of Bcl-2, decrease of DNA content, the release of cytochrome c into cytosol, translocation of AIF and DFF40 (CAD) onto nuclei, and activation of caspase-9, caspase-7, caspase-3, PARP and DFF45 (ICAD) whereas each single treated SCC25 cells did not show these patterns. Although the single treatment of $25{\mu}M$ HS-1200 and $4{\mu}g/ml$ cisplatin for 24 h did not induce apoptosis, the co-treatment of these reagents prominently induced apoptosis. Therefore our data provide the possibility that the combination therapy with HS-1200 and cisplatin could be considered as a novel therapeutic strategy for human squamous cell carcinoma.

Effects of Korean Zingiber mioga R. (Flower Buds and Rhizome) Extract on Memory (한국산 양하(꽃봉오리와 지하경)의 인지 기능 개선 효과)

  • Cho, Kyo-Hee;Oh, Myung-Sook;Kim, Hyo-Geun;Lee, Sun-Hee;Chung, Kun-Sub;Kim, Ae-Jung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.10
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    • pp.1519-1526
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    • 2014
  • This study investigated the biological activities and effects of Korean Zingiber mioga R. (flower buds and rhizome) on memory. The general composition, minerals, anti-oxidative activities, and AChE inhibitory effects were analyzed, and NORT (Novel object recognition test) and Y-Maze test in vivo were performed. The general contents (moisture, crude fat, crude protein, and crude ash; wet basis) of ZB (flower buds) were 91.96%, 0.15%, 1.99%, and 11.90%, respectively. The general contents (moisture, crude fat, crude protein, and crude ash; wet basis) of ZR (rhizome) were 75.21%, 0.53%, 2.20%, and 9.50%, respectively. The macro mineral contents (Ca, P, Na, and K) of ZB were 31.70 mg%, 15.20 mg%, 8.20 mg%, and 258.60 mg%, respectively. Inhibitory effects (IC50 value) of DPPH and ABTS radicals were higher with ZBD (flower buds water extract) than with ZBE (flower buds EtOH extract), ZRD (rhizome water extract) or ZRE (rhizome EtOH extract). AChE inhibitory effect of ZBD was higher and that of ZRD. NORT and Y-Maze test were performed with scopolamine-induced mice treated with ZBD and ZBE. In NORT, effects of ZBD and ZBE were similar to that of donepezil. In the Y-maze test, performances of ZBD and ZBE-treated mice were similar to that of the normal group. These results suggest that Korean Zingiber mioga R. has potential to be developed into a new functional food for cognition enhancement in the global food market.

Combination Gene Therapy of Herpes Simplex Virus Thymidine Kinase and Cytokines in Lung Cancer (폐암에서의 Herpes Simplex Virus Thymidine Kinase 유전자 치료와 Cytokine 유전자 치료의 복합요법)

  • Kim, Gye-Su;Park, Kyung-Ho;Seal, Ja-Young;Yoo, Chul-Gyu;Lee, Choon-Taek;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Sao
    • Tuberculosis and Respiratory Diseases
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    • v.51 no.2
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    • pp.135-146
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    • 2001
  • Background : One of the important mechanisms responsible for a tumor escaping the immune response is an absence of the tumor associated antigen (TAA) on the cancer cell surface. To overcome this, combination gene therapy using a herpes simplex thymidine kinase (HSTK) gene, prototype of drug sensitizing gene, was conducted to enhance T AA release by cell destruction, as well as the cytokine genes for immune cell attraction. Methods : We investigated whether or not transduction with the adenovirus-HSTK (Ad-HSTK) enhanced the sensitivity of Lewis lung carcinoma (LLC) to ganciclovir (GCV) and induced a bystander effect. A Tumor vaccine trial was performed using LLC with ad-HSTK$\pm$ad-GM-CSF$\pm$ad-IL-2 to determine if they exhibit some antitumor effect on established lung cancer xenografts. Results : LLC with ad-HSTK revealed a much higher sensitivity to ganciclovir (GCV). LLC transduced with ad-HSTK and/or ad-IL-2, ad-GM-CSF showed a lower in vivo tumorigenicity. In the treatment experiment, vaccination with LLC transduced with ad-HSTK, ad-IL-2, or ad-GM-CSF alone modestly suppressed the growth of an established tumor. Combined transduction with HSTK and GM-CSF induced stronger growth suppression of a established lung cancer, while HSTK and IL-2 combination transduction did not have any antitumor effect on individual transduction. Vaccination with LLC-HSTK-GM-CSF increased the infiltration of dendritic cells in the spleen. Conclusion : It was concluded that a tumor vaccine transduced with HSTK and GM-CSF induces strong antitumor immunity by activating the dendritic cells.

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Properties of the Silkworm (Bombyx mori) Dongchunghacho, a Newly Developed Korean Medicinal Insect-borne Mushroom: Mass-production and Pharmacological Actions (한국에서 개발된 곤충유래 약용버섯인 누에동충하초의 생산기술개발 및 약리학적 특성)

  • Lee, Sang Mong;Kim, Yong Gyun;Park, Hyean Cheal;Kim, Keun Ki;Son, Hong Joo;Hong, Chang Oh;Park, Nam Sook
    • Journal of Life Science
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    • v.27 no.2
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    • pp.247-266
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    • 2017
  • Cordyceps is a traditional Chinese medicinal herb well-known in China, Korea and Japan since B.C. 2,000. The original entomopathogenic fungus, Cordyceps sinensis belonging to the genus Cordyceps could not be found inside Korean peninsula due to the absence of the host insect for the corresponding entomogenous fungus. The development of artificial production methods of Korean type Cordyceps using the silkworm Bombyx mori as in vivo culture medium for the the entomopathogenic fungus Paecilomyces tenuipes is the first, and wonderful occasion in the research history of insect industry of this global world. The aim of this article is to review the historical research background, mass-production methods, and pharmacological effects of the silkworm-dongchunghacho (Paecilomyces tenuipes) which is a newly developed Korean medicinal insect-borne mushroom, and another non-insect-borne medicinal mushroom (Cordyceps militaris and Cordyceps pruinosa). Their biological actions include anti-tumor, immunostimulating, anti-fatigue, anti-stress, anti-oxidant, anti-aging, anti-diabetic, anti-inflammatory, anti-thrombosis, hypolipidaemic and insecticidal effects. The bioactive principles are protein-bound polysaccharides (hexose, hexosamin), cordycepin, D-manitol, acidic polysaccharide etc. Protein-bound polysaccharides and n-butanol fractions were demonstrated to show a significant anti-tumor activities but did not show a cytotoxicities. D-mannitol exhibited a significant prolongation of the life span in tumor bearing mice. Ergosterol did not show an efficient anti-tumor activity, but showed a significant phagocytosis enhancing activity. Anti-tumor activity of silkworm-dongchunghacho might be attributed to immuno-stimulating activities rather than cytotoxic effects [164]. Also this review comprises the breeding of Dongchunghacho varieties, optimization of culture conditions, improvement of learning and memory by Dongchunghacho, application of them as foods and chemical constituents.

Anti-osteoporotic Activity of Gojineumja Aqueous Extracts on the Ovariectomized Mice (난소적출 마우스에서 고진음자(固眞飮子) 물 추출물의 골다공증 개선 효과)

  • Cho, Su-Yun;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.31 no.4
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    • pp.16-38
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    • 2018
  • Objectives: The objective of this in vivo study is to observe the anti-osteoporotic activities of Gojineumja aqueous extracts (GJEJ) on the ovariectomized (OVX) mice as compared to those of risedronate sodium (RES). Methods: Thirty five days after bilateral OVX, GJEJ was orally administered, for 35 days once a day and then the changes on the body weight and gain during experimental periods, femur weights, bone mineral density (BMD), bone strength (failure load), mineral contents - calcium (Ca) and inorganic phosphorus (IP), histological profiles and histomorphometrical analyses at sacrifice were conducted with serum biochemistry - osteocalcin contents and bone specific alkaline phosphatase (BALP) activities. And the results of GJEJ were compared with RES orally administered OVX mice. Results: As a result of OVX, noticeable increase of body weight and gains and serum osteocalcin levels, decrease of serum BALP activities, femur weights, femur Ca and IP contents, BMD and strength were observed as compared to those of sham control mice, respectively. Also, the decrease of all histomorphometrical indices indicating the bone mass and structure, and the increase of indices about resorption were also detected in the femur of OVX control. However, these estrogen-deficient osteoporotic signs were significantly and dose-dependently inhibited by 35 days of continuous oral treatment of GJEJ, at dose levels of 500, 250 and 125 mg/kg, respectively. Especially, GJEJ 500 mg/kg showed favorable inhibitory activities against estrogen-deficient osteoporosis symptoms induced by OVX as comparable to those of RES 2.5 mg/kg. Conclusions: The results in this study suggest that oral administrations of GJEJ have clear dose-dependent favorable anti-osteoporotic activities in OVX mice.

Objective in Vivo Quantification of Emphysema by Thin-Section CT: Correlation with Physiologic Findings (고해상 전산화단층촬영을 이용한 폐기종의 정량적 분석: 폐기능 검사와의 비교)

  • Lee, Jee-Young;Lee, Kye-Young;Choi, Eun-Kyoung;Kim, Sang-Joon;Choi, Young-Hi
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.5
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    • pp.992-999
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    • 1998
  • Background: To correlate the emphysema score for quantification of the overall extent of emphysema in both lungs by CT with physiologic fingings and to get more objective and simple method to assess the extent of emphysema. Method: Thin-section CT and pulmonary function test(PFT) were performed in 17 patients with emphysema (all males, mean age, 62 years). Emphysema score was obtained as percentage of emphysematous lung area, dividing the total area of the emphysema(voxels with attenuation value less than -880, -900, -920HU, respectively) by the overall area of both lungs(voxels with attenuation value less than -400HU) with highlighting voxels using "Density mask" program. Emphysema score was calculated from whole lung(ESV) and 5 representative scans(ESR) using "Density mask", Visual emphysema score(ESV) was obtained by visual assessment from 5 representative scans. Correlation of these emphysema scores(ESW, ESR, ESV) and physiologic findings were performed, comparing the ESW with ESR and ESV. Results: ESW had correlation with DLCO(r=0.53-0.64) and $FEV_1/FVC$(r=0.42-0.57) among PFT parameters. ESR had good correlation with ESW and with PFT parameters as well. ESV did not correlate with PFT parameters except DLCO. Conclusion: CT quantification of emphysema using "density mask" correlated well with physiologic findings. To assess the severity of emphysema, both ESW and ESR are more reliable than ESV, and ESR is recommended in routine practice as it is objective, simple and reliable.

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The effect of different crystallization temperature of the hydroxyapatite coating produced by ion beam-assisted deposition on anodizing-treated titanium disks on human osteosarcoma cells (양극산화처리된 티타늄 표면에 이온빔보조증착방식을 이용한 수산화인회석 코팅시 소결온도의 차이가 조골세포에 미치는 영향)

  • Pae, Ah-Ran;Won, Hyun-Du;Lee, Richard Sung-Bok;Kim, Hyeong-Seob;Woo, Yi-Hyung
    • The Journal of Korean Academy of Prosthodontics
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    • v.49 no.4
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    • pp.333-340
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    • 2011
  • Purpose: The aim of this study was to study the effect of hydroxyapatite (HA) coating crystallinity on the proliferation and differentiation of human osteosarcoma cells. Materials and methods: Surface roughness of the titanium disks increased by anodizing treatment and then HA was coated using ion beam-assisted deposition (IBAD). HA coating was crystallized by heat-treated at different temperature ($100^{\circ}C$, $300^{\circ}C$, $500^{\circ}C$, $800^{\circ}C$). According to the temperature, disks were divided into four groups (HA100, HA300, HA500, HA800). With the temperature, crystallinity of the HA coating was different. Anodized disks were used as control group. The physical properties of the disk surface were evaluated by surface roughness tests, XRD tests and SEM. The effect of the crystallinity of HA coating on HOS cells was studied in proliferation and differentiation. HOS cells were cultured on the disks and evaluated after 1, 3, 5, and 7 days. Growth and differentiation kinetics were subsequently investigated by evaluating cell proliferation and alkaline phosphatase activity. Results: Regardless of the heat-treated temperature, there is no difference on the surface roughness. Crystallinity of the HA was appeared in the groups of HA500, HA800. HOS cells proliferation, ALP activity were higher in HA500 and HA800 group than HA100 and HA300. Conclusion: Within the results of this limited study, heat treatment at $500^{\circ}C$ of HA coating produced by IBAD has shown greater effect on proliferation and differentiation of HOS cells. It is considered that further in vivo study will be necessary.

ROS Scavenging Effect and Cell Viability of Opuntia humifusa Extract on Osteoblastic MC3T3-E1 Cells (천년초 추출물이 조골세포의 증식과 ROS소거능에 미치는 영향)

  • Hwang, Hyun-Jung;Jung, Bok-Mi;Kim, Mi-Hyang
    • Journal of Life Science
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    • v.21 no.12
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    • pp.1752-1760
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    • 2011
  • In this study, the effect of the Opuntiahumifusa extracts on proliferation, alkaline phosphatase (ALP) activity, collagen synthesis and ROS level of a cell was investigated using an osteoblast. Opuntiahumifusawas separated intoOpuntiahumifusapeel (OH-P), seed (OH-Se) and stem (OH-St).These were subjected to extraction by using hot water and ethanol. The proliferation of the MC3T3-E1 osteoblastic cells that were treated with OH-Se water extract were increased by approximately 120%. Regarding the effects of OH-Se on ALP activity, the $50{\mu}g/ml$ ethanol extract group showed the highest activity. The synthesis of collagen increased significantly in response to treatment with OH-Se water extract. The ROS scavenging effects of Opuntiahumifusawere investigated for involvement of oxidativedamage, cell culture and staining. Also, when OH-Se water extract $100{\mu}g/ml$ was added, the ROS level decreased by 54%. These results indicate that Opuntiahumifusa extracts have an anabolic effect on bone through the promotion of osteoblastic differentiation, suggesting that it could be used for the treatment of common metabolic bone diseases.

Gene Expression of Surfactant Protein B and C in Endotoxin and Thiourea Treated Rats (내독소 및 Thiourea 투여 후 Surfactant protein B와 C 유전자 발현의 비교 관찰)

  • Sohn, Dong Hyun;Sohn, Jang Won;Yoon, Ho Joo;Shin, Dong Ho;Park, Sung Soo
    • Tuberculosis and Respiratory Diseases
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    • v.54 no.5
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    • pp.510-521
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    • 2003
  • Background : The surfactant specific proteins, SP-B and SP-C are believed to be important regulators of the surfactant function and homeostasis. Since acute respiratory distress syndrome(ARDS) is usually viewed as the functional and morphological expression of a similar underlying lung injury caused by a variety of insults, and since abnormalities in the surfactant function have been described in ARDS, the authors investigated the different effects of endotoxin and thiourea on the accumulation of mRNA encoding SP-B and SP-C. Methods : Sprague-Dawley rats were given 5 mg/kg of an intraperitoneal endotoxin from Salmonella enteritidis and 3.5 mg/kg intraperitoneal thiourea and were sacrificed at different time periods. Results : 1. The SP-B mRNA levels 6 and 24 hours after the 5 mg/kg endotoxin treatment was significantly reduced by 26.1% and 50%, respectively(P<0.01, P<0.001). 2. The SP-B mRNA levels 24 hours after the 3.5 mg/kg thiourea treatment was reduced by 9.8% and 12.5%, respectively. 3. The SP-C mRNA levels 6 and 24 hours after the 5 mg/kg endotoxin treatment was significantly reduced by 38.7% and 53.6%, respectively(P<0.01, P<0.001). 4. The SP-C mRNA level 6 hours after the 3.5 mg/kg thiourea treatment was reduced by 22.8%(P<0.05). Conclusion : These results indicate that the differential regulation of the hydrophobic surfactant proteins in vivo is evident, and suggest that the hydrophobic surfactant proteins might be differentially regulated during lung injury at different time periods without altering the lung wet to dry ratios. The mechanism of these alternations at the different time periods and the different kinds of etiology remain to be determined.

Enhancement of Sensitivity of Human Lung Cancer Cell Line to TRAIL and Gefitinib by IGF-1R Blockade (폐암세포주에서 IGF-1R 억제를 이용한 TRAIL 및 gefitinib에 대한 감수성 증가를 위한 연구)

  • Lee, Yoon-Jin;Park, Mi-Young;Kang, Young-Ae;Kwon, Sung-Youn;Yoon, Ho-Il;Lee, Jae-Ho;Lee, Choon-Taek
    • Tuberculosis and Respiratory Diseases
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    • v.63 no.1
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    • pp.42-51
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    • 2007
  • Background: TRAIL is a cytokine that selectively induces apoptosis in various cancer cell lines. Gefitinib is new targeted drug applied in lung cancer that selectively inhibits EGFR tyrosine kinase. However, lung cancers have shown an initial or acquired resistance to these drugs. This study examined the effect of IGF-1R and its blockade on enhancing the sensitivity of lung cancer cell lines to TRAIL and gefitinib. Methods: Two lung cancer cell lines were used in this study. NCI H460 is very sensitive to TRAIL and gefitinib. On the other hand, A549 shows moderate resistance to TRAIL and gefitinib. The IGF-1R blockade was performed using adenoviruses expressing the dominant negative IGF-1R and shRNA to IGF-1R and AG1024 (IGF-1R tyrosine kinase inhibitor). Results: The adenovirus expressing dominant negative IGF-1R(950st) induced the increased expression of defective IGF-1R on the lung cancer cell surface, and the adenovirus-shIGF-1R effectively decreased the level of IGF-1R expression on cell surface. The genetic blockade of IGF-1R by the adenovirus-dnIGF-1R and AG1024 increased the sensitivity of A549 cells to TRAIL. The reduction of IGF-1R by transduction with ad-shIGF-1R also increased the sensitivity of the A549 cells to gefitinib. Conclusion: The blockade of IGF-1R through various mechanisms increased the sensitivity of the lung cancer cell line that was resistant to TRAIL and gefitinib. However, further studies using other cell lines showing acquired resistance as well as in vivo animal experiments will be needed.