• Title/Summary/Keyword: SD cell

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Isolation and morphological characterization of natural killer cell in the sprague-dawley(SD) rats (Sprague-dawley(SD) 랫드에서 natural killer cell의 분리 ·동정 및 형태적 특징)

  • Kang, Kyung-sun;Lee, Yong-soon
    • Korean Journal of Veterinary Research
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    • v.32 no.2
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    • pp.245-250
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    • 1992
  • This study was performed to demonstrate the presence of large granular lymphocyte(LGL) in Sprague-Dawley(SD) rats and morphologically observe NK cell and also establish the method of isolation of natural killer cell in SD rats. By percoll discontinuous density gradients centrifugation, highly enriched LGL population were shown to fraction 2(border line between 44.2% and 50.8%). LGL were shown to bind selectively to YAC1 mouse lymphoma cell. This fraction expressed very high NK cell cytolysis. Therefore, we thought that LGL have NK activity in SD rats. The Morphology of rat LGL is very similar to that of human LGL. These cells have an eccentric kidney-shaped nucleus. Their most distinctive feature was their cytoplasmic azurophilic granules. Another distinguishing feature of rat LGL was their high cytoplasmic : nuclear ratio. It was concluded that LGL played a role part in mediating natural killer activity in this species.

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Cytotoxicity of SD-994 from Artemisia argyi against L1210 Cells with Concomitant Induction of Antioxidant Enzymes (황해쑥 추출정제물 SD-994의 L1210암세포에 대한 세포독성과 항산화효소의 유발)

  • 정대영;하혜영;김안나;이승민;민태진;박시원
    • YAKHAK HOEJI
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    • v.44 no.3
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    • pp.213-223
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    • 2000
  • SD-994 was prepared from methanol extract of Artemisia argyi by stepwise purification of solvent partioning and silica gel chromatography. In the course of this purification, fractions obtained at each step were investigated for their cytotoxicities against L1210 cells. Fractions A~G prepared from chloroform fraction showed considerable cytotoxicities raging 40~90% against L1210 cells. Subfractions I~IX obtained from fraction A exhibited various cytotoxicities and subfraction I (SD-994) was found to be the most effective compound. $IC_{50}$ values of SD-994 were measured to be $0.5{\;}{\mu\textrm{g}}/ml and less than $0.05{\;}{\mu\textrm{g}}/ml against L1210 cells and normal lymphocytes, respectively: When SD-994 was added to L1210 cell as cytotoxic agent, significantly increased amount of superoxide ($O_2^-$) and dramatically augmented activities of superoxide dismutase (SOD), specially MnSOD and glutathione peroxidase (GPx) were observed according to the concentration and incubation time. Whereas, in case of normal lymphocytes under the same condition, cytotoxicities were not apparent and the generation of superoxide ($O_2^-$) or the activity changes of SOD and GPx were insignificant. These results together indicate that the cytotoxic action of SD-994 against L1210 cell may be achieved via necrosis and/or apoptosis induced by reaction oxygen species which could not probably be completely abolished even by drastically increased antioxidant enzymes, SOD and GPx activities.

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Hematological Reference Values in the Healthy Adults (건강성인의 혈구 참고치 산정)

  • Kim, Young-Jin;Hyun, Myung-Soo;Lee, Hyun-Woo
    • Journal of Yeungnam Medical Science
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    • v.8 no.1
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    • pp.154-165
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    • 1991
  • To establish the hematological reference values in the healthy adults visited our hospitals, following examination were done on 2823 persons by Coulter Counter Model S-plus II ; white blood cell count: (WBC), red blood cell count(RBC), hemoglobin(Hb), hematocrit(Hct), mean corpuscular volume (MCV), mean corpuscular hemoglobin(MCH), mean corpuscular hemoglobin concentration(MCHC), red cell distribution width(RDW), platelet, plateletcrit, mean platelet volume(MPV) and platelet distribution width(PDW). The following results are obtained. 1) Male, mean value of WBC ; $6,800{\pm}2,680(2SD)/{\mu}l$ Female, mean value of WBC ; $5,950{\pm}2,380(2SD)/{\mu}l$ 2) Male, mean value of RBC ; $428{\pm}60(2SD){\times}10^4/{\mu}l$ Female, mean value of WBC ; $415{\pm}56(2SD){\times}10^4/{\mu}l$ 3) Male, mean value of Hb ; $15.4{\pm}1.8(2SD)g/dL$ Female, mean value of Hb ; $13.0{\pm}1.6(2SD)g/dL$ 4) Male, mean value of Hct ; $45.3{\pm}5.0(2SD)%$ Female, mean value of Hct ; $38.2{\pm}4.6(2SD)%$ 5) Male, mean value of MCV ; $93.8{\pm}5.8(2SD)fL$ Female, mean value of MCV ; $92.2{\pm}7.4(2SD)fL$ 6) Male, mean value of MCH ; $31.8{\pm}2.2(250)pg$ Female, mean value of MCH ; $31.4{\pm}2.8(2SD)pg$ 7) Male, mean value of MCHC ; $34.0{\pm}1.2(2SD)%$ Female, mean value of MCHC ; $33.9{\pm}1.2(2SD)%$ 8) Male, mean value of RDW ; $12.7{\pm}1.0(2SD)%$ Female, mean value of RDW ; $12.6{\pm}1.4(2SD)%$ 9) Male, mean value of Platelet ; $242.9{\pm}87.8(2SD){\times}10^3/{\mu}l$ Female, mean value of Platelet ; $242.2{\pm}89.0(2SD){\times}10^3/{\mu}l$ 10) Male, mean value of Plateletcrit ; $0.201{\pm}0.076(2SD)%$ Female, mean value of Plateletcrit ; $0.204{\pm}0.076(2SD)%$ 11) Male, mean value of MPV ; $8.20{\pm}1.70(2SD)fl$ Female, mean value of MPV ; $8.36{\pm}1.82(2SD)fl$ 12) Male, mean value of PDW ; $16.1{\pm}0.8(2SD)%$ Female, mean value of PDW ; $16.0{\pm}0.8(2SD)%$.

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Impact of Co-transfection with Livin and Survivin shRNA Expression Vectors on Biological Behavior of HepG2 Cells

  • Xu, Wei;Chang, Hong;Qin, Cheng-Kun;Zhai, Yun-Peng
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.9
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    • pp.5467-5472
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    • 2013
  • Objective: To construct short hairpin RNA (shRNA) eukaryotic expression vectors targeting Livin and Survivin genes, and to explore the impact of co-transfection of Livin and Survivin shRNA expression vectors on the biological behavior of HepG2 cells. Methods: shRNA eukaryotic expression vectors pSD11-Livin and pSD11-Survivin were designed and constructed then transfected into HepG2 cells separately or in combination. mRNA and protein expression in transfected cells was assessed by quantitative fluorescence PCR and Western blotting, respectively. Cell proliferation was measured by MTT assay and cell apoptosis by TUNEL assay. Results: The Livin and Survivin shRNA eukaryotic expression vectors were successfully constructed and transfected into HepG2 cells. The relative mRNA expression levels of Livin and Survivin in HepG2 cells co-transfected with pSD11-Livin and pSD11-Survivin were $0.12{\pm}0.02$ and $0.33{\pm}0.13$, respectively, which was significantly lower than levels in cells transfected with either pSD11-Livin or pSD11-Survivin (P<0.05). The relative protein expression levels of Livin and Survivin in the co-transfected cells were also significantly decreased compared to single-transfection (P<0.05). The inhibition rate of cell growth in the co-transfection group was higher than that in the single-transfection groups at 48 h, 60 h, or 72 h after transfection (P<0.01). The apoptotic rate increased to the greatest extent in the co-transfection group relative to any other group (P<0.05). Conclusions: Co-transfection with pSD11-Livin and pSD11-Survivin was more efficient than transfection with either vector alone in reducing the mRNA and protein expression of Livin and Survivin genes in HepG2 cells. Co-transfection also inhibited the proliferation of transfected cells more than the other groups, and induced cellular apoptosis more effectively.

White Matter Damage and Hippocampal Neurodegeneration Induced by Permanent Bilateral Occlusion of Common Carotid Artery in the Rat: Comparison between Wistar and Sprague-Dawley Strain

  • Kim, Seul-Ki;Cho, Kyung-Ok;Kim, Seong-Yun
    • The Korean Journal of Physiology and Pharmacology
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    • v.12 no.3
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    • pp.89-94
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    • 2008
  • In order to reproduce chronic cerebral hypoperfusion as it occurs in human aging and Alzheimer's disease, we introduced permanent, bilateral occlusion of the common carotid arteries (BCCAO) in rats (Farkas et al, 2007). Here, we induced BCCAO in two different rat strains in order to determine whether there was a strain difference in the pathogenic response to BCCAO. Male Wistar and Sprague-Dawley (SD) rats (250-270 g) were subjected to BCCAO for three weeks. Kluver-Barrera and cresyl violet staining were used to evaluate white matter and gray matter damage, respectively. Wistar rats had a considerably higher mortality rate (four of 14 rats) as compared to SD rats (one of 15 rats) following BCCAO. Complete loss of pupillary light reflex occurred in all Wistar rats that survived, but loss of pupillary light reflex did not occur at all in SD rats. Moreover, BCCAO induced marked vacuolation in the optic tract of Wistar rats as compared to SD rats. In contrast, SD rats showed fewer CA1 hippocampal neurons than Wistar rats following BCCAO. These results suggest that the neuropathological process induced by BCCAO takes place in a region-specific pattern that varies according to the strain of rat involved.

Serum Amyloid A as an Independent Prognostic Factor for Renal Cell Carcinoma - A Hospital Based Study from the Western Region of Nepal

  • Mittal, Ankush;Poudel, Bibek;Pandeya, Dipendra Raj;Gupta, Satrudhan Pd;Sathian, Brijesh;Yadav, Shambhu Kumar
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.5
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    • pp.2253-2255
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    • 2012
  • Objective: The objective of our present study was to assess the role of serum amyloid A (SAA) in stages and prognosis of renal cell carcinoma. Material and Methods: It was a hospital based retrospective study carried out in the Department of Medicine and Biochemistry of Manipal Teaching Hospital, Pokhara, Nepal between $1^{st}$ January 2008 and $31^{st}$ December 2011. The variables collected were SAA, CRP. Approval for the study was obtained from the institutional research ethical committee. Quantitative analysis of human SAA and C-reactive protein (CRP) was performed by radial immune diffusion (RID) assay for all cases. Results: Of the 422 total cases of renal cell carcinoma, 218 patients had normal and 204 abnormal SAA. SAA levels were grossly elevated in T3 stage ($122.3{\pm}SD35.7$) when compared to the mean for the T2 stage ($84.2{\pm}SD24.4$) (p value: 0.0001). Similarly, SAA levels were grossly elevated in M1 stage ($190.0{\pm}SD12.7$) when compared to the M0 stage ($160.9{\pm}SD24.8$) (p: 0.0001). There was no significant association with elevated CRP levels ($209.1{\pm}SD22.7$, normal $199.0{\pm}SD19.5$). Conclusion: The validity of SAA in serum as being of independent prognostic significance in RCC was demonstrated with higher levels in advanced stage disease.

The Effect of Suhwagije-tang Distillate on the Immune Activity of Spleen Cells of Aged Rats (수화기제탕(水火旣濟湯) 증류액이 노화 흰쥐의 비장세포 면역활성에 미치는 영향)

  • Choi, Jae-Song;Bae, Na-Young;Ahn, Teak-Won
    • Journal of Sasang Constitutional Medicine
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    • v.23 no.3
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    • pp.374-390
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    • 2011
  • 1. Objectives: The purpose of this study is to investigate the effect of Suhwagije-tang(SGT) distillate on the immune activity of spleen cells of aged SD rats. 2. Methods: We used 10, 50, 72 weeks old SD rats in this study. Spleen cells from SD rats were stimulated with ConA and treated with 1% Vitamin C(Vit.C) or Suhwagijetang distillate(SGT). After 24 hours, the concentrations of IL-2, IL-4, IL-10, IFN-${\gamma}$ in the cell culture supernatant were measured by ELISA. 3. Results and Conclusions 1) At all concentration of SGT distillate, survival rates of liver cells were higher than the control group. In addition, 50% SGT distillate group's cell survival rates were significantly higher than other groups. 2) In 10 weeks SD rats(SGT group), the concentration of IL-2 significantly decreased in comparison with ConA group, Vit.C group. In 52 weeks SD rats(SGT group), the concentration of IL-2 significantly decreased in comparison with ConA group. 3) In 10, 52 weeks SD rats(SGT group), the concentration of IL-4 significantly decreased in comparison with ConA group. 4) In 10 weeks SD rats(SGT group), the concentration of IL-10 significantly decreased in comparison with ConA group. And in 72 weeks SD rats(SGT group), the concentration of IL-10 significantly increased in comparison with Vit.C group. 5) In 52, 72 weeks SD rats(SGT group), the concentration of IFN-${\gamma}$ significantly decreased in comparison with 10 weeks SD rats(SGT group). These results suggest that Suhwagije-tang(SGT) distillate has the effect of increasing the immune activity of spleen cells of aged SD rats.

Effect of the low level light irradiation to NTacSam:SD tissue cell culture (NTacSam:SD의 조직세포 배양에 저출력 광원의 효과)

  • Kim, Tae-Gon;Kim, Toung-Pyo;Park, No-Bong;Lee, Ho-Sic;Park, Yong-Pil;Cheon, Min-Woo
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2009.06a
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    • pp.423-423
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    • 2009
  • Currently, lasers are one of the most popular light sources in use for medical treatment. Many studies on low power lasers are being done in cell culture or through animal tests and most report different findings, making it difficult to verify their true effects. There are shifts in trends of studies from laser and LED that are expensive and generate heat problem to LED that are economically effective and safe. Its near infrared rays can penetrate deep into skin or muscle, up to 23 cm, without causing thermal damage or impairing neighboring tissues. This study verified the performance and effectiveness of an LED irradiator that was designed to emit similar wavelengths to that of a laser and thus could be used instead of a low level laser therapy in experiments on animals. And then, each experiment was performed to irradiation group and non-irradiation group for NTacSam:SD tissue cells. MIT assay method was chosen to verify the cell increase of two groups and the effect of irradiation on cell proliferation was examined by measuring 590nm transmittance of ELISA reader. As a result, the cell increase of NTacSam:SD tissue cells was verified in irradiation group as compared to non-irradiation group. The fact that specific wavelength irradiation has an effect on cell vitality and proliferation is known through this study.

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Effect of Silver sulfadiazine on the Skin Cell Proliferation and Wound Healing Process in Hairless Mouse 2nd degree Burn Model (설파디아진은의 피부세포 증식 및 화상모델에 있어서의 상처치유과정에 미치는 영향)

  • Cho, Ae-Ri
    • Journal of Pharmaceutical Investigation
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    • v.32 no.2
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    • pp.113-117
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    • 2002
  • Cyto-toxic effect of silver sulfadiazine (Ag-SD) on keratinocytes and its implication on wound healing process were investigated in $2^{nd}$ degree bum hairless mouse model. As a dermal model, HaCat (immortalized keratinocytes) monolayer culture in DMEM with 10% FBS was used. Cyto-toxicity of Ag-SD was estimated by measuring the cell viability using neutral red assay after adding the drug. The $2^{nd}$ degree bum was prepared on hairless mouse back skin (1 cm diameter) and dressings with Ag-SD were applied for 96 hr. The process of re-epithelialization and the presence of inflammatory cells were investigated and histology with Hematoxylin-Eosin staining was performed. Ag-SD displayed highly cyto-toxic effect on cultured HaCat cells in a concentration dependent manner $(1-100\;{\mu}g/mL)$. Topical application of Ag-SD (2%) could control the infection: no inflammatory cells were observed in histology. However the cyto-toxic effect of Ag-SD on skin cells induced the impairment in epidermal regeneration.

Effect of the Physiologically Active Compounds in Phlomidis Radix on Cell Cycle Regulation in Human Gingival Fibroblasts (속단의 생리활성성분이 치은섬유아세포의 세포주기조절에 미치는 영향)

  • You, Suk-Joo;Jang, Kil-Young;Yoon, Ho-Sang;Choi, Ho-Chul;Sung, Ki-Jong;Kim, Hyun-A;Pi, Sung-Hee;Shin, Hyung-Shik;You, Hyung-Keun
    • Journal of Periodontal and Implant Science
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    • v.35 no.1
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    • pp.87-98
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    • 2005
  • The purpose of present study was to investigate the effects of physiologically active compound (SD62-122) from Phlomidis Radix on the cell cycle progression and its molecular mechanism in human gingival fibroblasts(HGFs). For this purpose, fibroblasts were isolated and cultured from excisioned gingiva during crown lengthening procedure in healthy adult. The following parameter were evaluated that there are cell number counting, MIT assay, cell cycle progression, western blot analysis. The cell number and MIT assay of primary cultured fibroblast was not increased at 2 days but significant increased compare to negative control at 3days(p<0.05). S phase was increased and G1 phase decreased in both $10^{-8}M$ and $10^{-9}M$ of SD62-122 in cell cycle analysis. The cell cycle regulation protein levels of Cyclin $D_1$, Cyclin E, cdk 2, cdk 4 and cdk 6 were increased compare to control in both $10^{-8}M$ and $10^{-9}M$ of SD62-122. The protein levels of p21 and p53 were decreased compare to control, but the level of pRb was not changed compare to control in $10^{-9}M$ of SD2-122. These results suggested that physiologically active compound (SD62-122) isolated from Phlomidis Radix increases the cell proliferation and cell cycle progression in HGFs, which is linked to increased cell cycle regulation protein levels of Cyclin $D_1$, Cyclin E, cdk 2, cdk 4 and cdk 6, and decreased the levels of p21, p53.