• Title/Summary/Keyword: Tissue growth

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Improvement of Liver Function and Suppressed Lipid Peroxidation of Extract from Ginseng Folium and Stem in Acute $CCl_4$ Intoxicated Rats ($CCl_4$로 급성 간손상을 유도한 백서에서 인삼엽과 경추출물의 간기능 개선과 항산화 작용)

  • Lee Min Kyung;Park Sung Hye;Seo Eui Suok;Kim Ki Young
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.5
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    • pp.1485-1489
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    • 2004
  • Panax ginseng is the one of best famous phytochemical plant in the world and it's various positive effects such as antioxidant, regulation of immunity are very well known. In this study, we investigated primary the cell viability and morphological change and secondary an antioxidative effect and liver function improvement of extract from Ginseng folium and stem in CCl4 intoxicated rats. The NCTC cell line were used for cell viability and sirius red staining before the animal experiment. The female Sprague-Dawley rats (90-100g) were divided into 3 groups (Normal, AC: CCl₄ treated group, GFS: CCl₄+ extract of Ginseng folium and stem treated group) and acute liver damage was developed by one time administration of CCl₄ mixture (0.5㎖/rat). The liver tissue and sera were collected and used for quantitative measurement of enzyme activity (AST, ALT, ALP, BUN), MDA and Hyp. As a result, cell viability in GFS treated group (in concentration of 3.33-33.33㎎ GFS/200㎕ medium) was 180.9-241.0% significantly and dose dependently higher than in control group. And potential state of cell growth and differentiation and no criteria of cytoplasm lysis and nucleus breaking were observed in control and GFS group. The parameters of liver function (AST and ALP) in sera of GFS group showed significantly 93% and 67.6% lower than AC group (p<0.005-0.05). And the level of ALT and BUN showed fast similar in AC group and GFS group. The concentration of MDA in liver was decreased 576.5% significantly in GFS group when compared with AC group (p<0.005). The content of Hyp in GFS group is merely lower than in AC group. In conclusion, the water extract of Ginseng folium and stem such as Ginseng radix may be possessed the antioxidative effect and improvement of liver function in CCl₄ intoxicated rats.

Bio-Conjugated Polycaprolactone Membranes: A Novel Wound Dressing

  • Cai, Elijah Zhengyang;Teo, Erin Yiling;Jing, Lim;Koh, Yun Pei;Qian, Tan Si;Wen, Feng;Lee, James Wai Kit;Hing, Eileen Chor Hoong;Yap, Yan Lin;Lee, Hanjing;Lee, Chuen Neng;Teoh, Swee-Hin;Lim, Jane;Lim, Thiam Chye
    • Archives of Plastic Surgery
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    • v.41 no.6
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    • pp.638-646
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    • 2014
  • Background The combination of polycaprolactone and hyaluronic acid creates an ideal environment for wound healing. Hyaluronic acid maintains a moist wound environment and accelerates the in-growth of granulation tissue. Polycaprolactone has excellent mechanical strength, limits inflammation and is biocompatible. This study evaluates the safety and efficacy of bio-conjugated polycaprolactone membranes (BPM) as a wound dressing. Methods 16 New Zealand white rabbits were sedated and local anaesthesia was administered. Two $3.0{\times}3.0cm$ full-thickness wounds were created on the dorsum of each rabbit, between the lowest rib and the pelvic bone. The wounds were dressed with either BPM (n=12) or Mepitel (n=12) (control), a polyamide-silicon wound dressing. These were evaluated macroscopically on the 7th, 14th, 21st, and 28th postoperative days for granulation, re-epithelialization, infection, and wound size, and histologically for epidermal and dermal regeneration. Results Both groups showed a comparable extent of granulation and re-epithelialization. No signs of infection were observed. There was no significant difference (P>0.05) in wound size between the two groups. BPM (n=6): $8.33cm^2$, $4.90cm^2$, $3.12cm^2$, $1.84cm^2$; Mepitel (n=6): $10.29cm^2$, $5.53cm^2$, $3.63cm^2$, $2.02cm^2$; at the 7th, 14th, 21st, and 28th postoperative days. The extents of epidermal and dermal regeneration were comparable between the two groups. Conclusions BPM is comparable to Mepitel as a safe and efficacious wound dressing.

Establishment of an Allo-Transplantable Hamster Cholangiocarcinoma Cell Line and Its Application for In Vivo Screening of Anti-cancer Drugs

  • Puthdee, Nattapong;Vaeteewoottacharn, Kulthida;Seubwai, Wunchana;Wonkchalee, Orasa;Keawkong, Worasak;Juasook, Amornrat;Pinloar, Somchai;Pairojkul, Chawalit;Wongkham, Chaisiri;Okada, Seiji;Boonmars, Thidarut;Wongkham, Sopit
    • Parasites, Hosts and Diseases
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    • v.51 no.6
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    • pp.711-717
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    • 2013
  • Opisthorchis viverrini (O. viverrini) is a well-known causative agent of cholangiocarcinoma (CCA) in humans. CCA is very resistant to chemotherapy and is frequently fatal. To understand the pathogenesis of CCA in humans, a rodent model was developed. However, the development of CCA in rodents is time-consuming and the xenograft-transplantation model of human CCA in immunodeficient mice is costly. Therefore, the establishment of an in vivo screening model for O. viverrini-associated CCA treatment was of interest. We developed a hamster CCA cell line, Ham-1, derived from the CCA tissue of O. viverrini-infected and N-nitrosodimethylamine-treated Syrian golden hamsters. Ham-1 has been maintained in Dulbecco's Modified Essential Medium supplemented with 10% fetal bovine serum for more than 30 subcultures. These cells are mostly diploid (2n=44) with some being polyploid. Tumorigenic properties of Ham-1 were demonstrated by allograft transplantation in hamsters. The transplanted tissues were highly proliferative and exhibited a glandular-like structure retaining a bile duct marker, cytokeratin 19. The usefulness of this for in vivo model was demonstrated by berberine treatment, a traditional medicine that is active against various cancers. Growth inhibitory effects of berberine, mainly by an induction of G1 cell cycle arrest, were observed in vitro and in vivo. In summary, we developed the allo-transplantable hamster CCA cell line, which can be used for chemotherapeutic drug testing in vitro and in vivo.

Anomalous Structure of Somatic Embryos Developed from Leaf Explant Cultures of Angelica gigas Nakai

  • Cho, Duck-Yee;Lee, Eun-Kyong;Soh, Woong-Young
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.1
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    • pp.1-5
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    • 1998
  • This study describes the effect of the growth regulators such as 2,L-D and BA, on the structural abnormalities of somatic embryos derived from leaf explants of Angelica gigas Nakai, Also, the relationship between the cotyledon number of a somatic embryo and its germinability is explored. Embryogenic calli were selected from calli formed on explants cultured on MS solid basal medium supplemented with 0.5mg/L 2,4-D, 1mg/L 2,4-D, 1mg/L plus 0.1mg/L BA, and 1 mg/L 2,4-D plus 0.5mg/L BA. Cotyledonary abnormalities were observed in somatic embryos which were developed from embryogenic calli cultured on MS medium containing 1mg/L 2,4-D for 8 weeks and then subcultured on 2,4-D free MS medium for 3 weeks. The frequency of abnormalities was as follows: 22.8% one cotyledon, 42.5% two cotyledons, 16.8% three cotyledons, 7.8% four cotyledons, 1.8% five cotyledons, and 8.2% jar shaped cotyledon. In addition, ABA treatment indicated an improvement of the somatic embryo with normal cotyledon (65.3%). ABA was important role to the high production of normal somatic embryos. Two cotyledon embryos showed germinability 77.8%. However the germinability of somatic embryos with anomalous cotyledons was prominently low: One cotyledon, 62.5%; three cotyledons, 43.3%; four cotyledons, 60%; five cotyledons, 50% and jar shaped cotyledon, zero%. Thus, germinability was essentially, inversely proportional to cotyledon number.

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Superoxide Dismutase Activity in Suspension Cultured Cells of Tomato (Lycopersicon esculentum Mill) (토마토(Lycopersicon esculentum Mill) 현탁배양세포에서 Superoxide Dismutase 활성)

  • 유순희;허경혜;권석윤;이행순;방재욱;곽상수
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.1
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    • pp.57-61
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    • 1997
  • We investigated changes in the superoxide dismutase (SOD) activity and SOD isoenzyme pattern in suspension cultures of tomato (Lycopersicon esculentum), which were compared with those of intact tomato plants. two grams (fr wt) of cells subcultured at 15-day intervals were inoculated into 50 mL MS medium containing l mg/L 2,4-D and 30 g/L sucrose in a 300 mL flask and maintained at $25^{\circ}C$ in the dark (100 rpm). The cell growth reached a maximum at 20 days after subculture (DAS), followed by a rapid decrease with further cultures. The cell colour changed from white to black from 23 DAS. The intracellular SOD activity (units/g cell dry wt) was significantly increased from 23 DAS and reached a maximum at 28 DAS (52,400 units), followed by a decrease with further cultures, whereas the extracellular SOD activity showed a maximum at 25 DAS (27,800 units/50 mL medium). The total SOD activity per flask showed a maximum at 25 DAS (35,700 units), in which the extracellular SOD activity occupied about 75%. The tomato cultured cells had four SOD isoenzymes and their patterns were well correlated with SOD activity without a qualitative change during the cell cultures. The intact tomato plants had an additional CuZnSOD isoenzyme, showing the different isoenzyme patterns from cultured cells.

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Expression of Recombinant Erythropoietin Gene in Transgenic Tobacco Plant (형질전환 담배 식물체에서 재조합 erythropoietin 유전자의 발현)

  • CHOI, Jang Won;PARK, Hee Sung
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.1
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    • pp.63-69
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    • 1997
  • Erythropoietin (EPO) is a glycoprotein that mediates the growth and differentiation of erythroid progenitors. In order to produce recombinant human erythropoietin in tobacco plant, the EPO genomic DNA (5.4 kb) was cloned into plant expression vectors, pBI$\Delta$GUS121, pBD$\Delta$GUS121 and pPEV-1, and introduced in Nicotiana tabacum (var. Xanthi) via Agrobacterium tumefaciens-mediated transformation. After selection on MS media containing kanamycin (Km), 10 Km-resistant plants were obtained per each construct. The correct integration of EPO genomic DNA in the genome of transgenic plant was confirmed by polymerase chain reaction (PCR). Northern blot showed that transcripts of 1.8 kb length were produced in leaves of the plants, but there was no difference of mRNA amount according to promoter number and 5'-untranslated sequence (UTS). The proteins obtained from leaves of transgenic plants were immunologically detected by Western blot using rabbit anti-human EPO polyclonal antibody. The expressed protein appeared as smaller band of apparent mass of 30 kDa as compared to the EPO protein from human urine (37 kDa), suggesting that the modification (glycosylation) system in tobacco plant might be different from that of mammalian cells.

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Development of Pollen-Derived Embryos and Ploidy Level of Their Regenerated Plants in Paeonia ladctiflora Pall. (작약(Paeonia lactiflora Pall.) 화분에서 유래된 배의 발달과 분화식물체의 배수성)

  • 손재근;김광수;김경민
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.4
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    • pp.215-220
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    • 1994
  • Pollen-derived embryos cultured on the hormone-free medium showed a low germination frequency (12.5%) and poor growth response after germination. The greatest frequency of germination (81.3%) was obtained from the embryos cultured on medium with 0.3mg/L GA$_3$.The greatest frequency of generation (81.3) was obtained from embryos cultured on medium with 0.3mg/L GA$_3$. The embryos precultured for 20 days on medium with 0.3mg/L GA$_3$were transferred to the medium with various combination of hormones such as IAA, kinetin, zeatin, 6-benzylaminopurin (BA) and Gh$_3$. The germination frequency of cotyledonary stage embryos showed above 72% on media with all of the hormonal combinations, but the embryos germinated on medium with 2mg/L BA or 0.1mg/L kinetin and 0.3mg/L GA$_3$ developed more vigorously into plantlets than those of other hormonal combinations. Torpedo-stage embryos cultured on medium with 0.3 mg/L Gh$_3$ were pretreated for 8 weeks at 2-week intervals at 4$^{\circ}C$, The germination frequency of the cold-preheated embryos increased with the increment of pretreatment period from 2 to 8 weeks. The greatest frequency of germination (73.3%) was obtained from the embryos pretreated for 8 weeks at 4$^{\circ}C$. The chromosomes of the root-tip cells of W plane grown for 40 days after germination were observed. Most of the regenerated plants were haploid (55.8%) or diploid (315%), but triploid (1.3%), tetraploid (5.2%), or aneuploid (6.5%) were also detected among them.

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Characteristics of Bed-log of Shiitake Damaged by Bjerkandera adusta and Antagonism between These Two Fungi (줄버섯 피해 표고골목의 특성 및 표고균과의 대치배양)

  • Bak, Won-Chull;Lee, Bong-Hun;Park, Young-Ae;Kim, Hyun-Seok
    • The Korean Journal of Mycology
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    • v.39 no.1
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    • pp.44-47
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    • 2011
  • A harmful fungus occurred seriously in bed-log of shiitake(Lentinula edodes) in Jangheung-Gun, Korea. The fungus was identified as Bjerkandera adusta by its morphology and ITS(Internal Transcribed Spacer) analysis. The fungus was reported as causal agent of stem-rot of Populus euramericana in Korea, but not reported in bed-log of shiitake until this notification. Thus, studies were made to investigate inside condition of bed-log of shiitake damaged by B. adusta, physiological characteristics of B. adusta and antagonism between these two fungi. First of all, B. adusta is white-rotting fungus like shiitake and wood-rotting condition is similar to that of shiitake. But, there are a lot of small spots in damaged wood tissue under bark which are not seen in case of shiitake. Optimal temperature for mycelial growth of B. adusta is ca. $30^{\circ}C$ while that of shiitake is ca. $25^{\circ}C$. When confrontation cultures were made between these two fungi under $15^{\circ}C$, $20^{\circ}C$, $25^{\circ}C$ and $30^{\circ}C$, B. adusta has antagonistic ability against shiitake in all the temperatures. From the results of experiments, if the bed-logs of shiitake are exposed to high temperature, there should be mass propagation of B. adusta, and shiitake mycelia will be seriously injured by the fungus. Therefore, to prevent the damage by B. adusta, it is needed to grow the mycelia of shiitake fast in the bed-log, and to avoid exposure of the bed-log to high temperature in summer.

In Vitro Development of Somatic Cell Nuclear Transfer Embryo Treated with Flavonoid and Production of Cloned Jeju Black Cattle (플라보노이드 처리된 체세포 핵이식 배아의 체외 발달 및 제주흑우 복제 소 생산)

  • Kim, Eun-Young;Kim, Yeon-Ok;Kim, Jae-Youn;Park, Min-Jee;Park, Hyo-Young;Han, Young-Joon;Mun, Seong-Ho;Oh, Chang-Eon;Kim, Young-Hoon;Lee, Sung-Soo;Ko, Moon-Suck;Park, Se-Pill
    • Reproductive and Developmental Biology
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    • v.34 no.3
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    • pp.127-134
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    • 2010
  • This study was to investigate the effect of flavonoid treatment on in vitro development of bovine somatic cell nuclear transfer (SCNT) embryos, and their pregnancy and delivery rate after embryo transfer into recipient. In experiment 1, to optimize the flavonoid concentration, parthenogenetic day 2 ($\geq$ 2-cell) embryos were cultured in 0 (control), 1, 10 and $20\;{\mu}M$ flavonoid for 6 days. In the results, in vitro development rate was the highest in $10\;{\mu}M$ flavonoid group (57.1%) among treatment groups (control, 49.5%; $1\;{\mu}M$, 54.2%; $20\;{\mu}M$, 37.5%), and numbers of total and ICM cells were significantly (p<0.05) higher in $10\;{\mu}M$ flavonoid group than other groups. We found that $10\;{\mu}M$ flavonoid treatment can significantly (p<0.05) decrease the apoptotic index and derive high expression of anti-oxidant, anti-apoptotic, cell growth and development marker genes such as Mn-SOD, Survivin, Bax inhibitor, Glut-5, In-tau, compared to control group. In experiment 2, to produce the cloned Jeju Black Cattle, beef quality index grade 1 bull somatic cells were transferred into enucleated bovine MII oocytes and reconstructed embryos were cultured in $10\;{\mu}M$ flavonoid added medium. When the in vitro produced day 7 or 8 SCNT blastocysts were transferred into a number of recipients, $10\;{\mu}M$ flavonoid treatment group presented higher pregnancy rate (10.2%, 6/59) than control group (5.9%, 2/34). Total three cloned Jeju Black calves were born. Also, two cloned calves in $10\;{\mu}M$ flavonoid group were born and both were all healthy at present, while the one cloned calf born in control group was dead one month after birth. In addition, when the result of short tandem repeat marker analysis of each cloned calf was investigated, microsatellite loci of 11 numbers matched genotype between donor cell and cloned calf tissue. These results demonstrated that the flavonoid addition in culture medium may have beneficial effects on in vitro and in vivo developmental capacity of SCNT embryos and pregnancy rate.

Effect of Eupatorium japonicum Extract on the Metastasis, Invasion and Adhesion of MDA-MB-231 Human Breast Cancer Cells (등골나물 추출물이 인간의 유방암세포인 MDA-MB-231 세포의 이동, 침윤 및 부착에 미치는 영향)

  • Woo, Eun-Young;Park, So-Young;Kwon, Soo-Jin;Kwon, Gyoo-Taik;Kim, Jong-Dae;Lim, Soon-Sung;Yoon, Jung-Han
    • Korean Journal of Food Science and Technology
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    • v.43 no.2
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    • pp.213-219
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    • 2011
  • The metastatic effect of Eupatorium japonicum extract (EJE) on MDA-MB-231 human breast cancer cells was investigated. MDA-MB-231 cells were treated with various concentrations of EJE (0, 5, 10 and $20{\mu}g/mL$). EJE inhibited cell migration, invasion and adhesion of MDA-MB-231 cells in dose-dependent manners. Gelatin zymography exhibited that EJE significantly down regulated secretion of matrix metalloproteinase (MMP)-9 and MMP-2. EJE decreased the protein levels of tissue inhibitor of metalloproteinase (TIMP)-1 but increased TIMP-2 levels. Additionally, EJE reduced the protein and mRNA levels of urokinase-type plasminogen activator (uPA), vascular endothelial growth factor (VEGF) and intercellular adhesion molecule (ICAM). In several solvent fractions of EJE, the hexane fraction markedly decreased MDAMB-231 cell migration. Thus, these finding suggest that EJE may be a potential antimetastatic agent, which can considerably inhibit the metastatic and invasive capacity of breast cancer cells.