• Title/Summary/Keyword: Cdc6

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Effects of FBS(Fetal Bovine Serum) and pFF(Porcine Follicular Fluid) on In Vitro Maturation and Development of Porcine Parthenogenetic and Nuclear Transfer Embryos

  • Moon, Hyo-Jin;Shim, Joo-Hyun;Hwang, In-Sun;Park, Mi-Rung;Kim, Dong-Hoon;Ko, Yeoung-Gyu;Park, Choon-Keun;Im, Gi-Sun
    • Reproductive and Developmental Biology
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    • v.33 no.2
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    • pp.85-91
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    • 2009
  • In this study, in vitro maturation system using fetal bovine serum (FBS) or porcine follicular fluid (pFF) was investigated to produce comparable oocytes to those derived from in vivo. Control group of oocytes was cultured in TCM 199 supplemented with 0.1% polyvinyl alcohol (PVA). Other three groups of oocytes were cultured in TCM 199 supplemented with 10% FBS, 10% pFF or 5% FBS + 5% pFF, respectively. After 44 h maturation, oocytes with the first polar body were activated with two electric pulses (DC) of 1.2 kv/cm for 30 ${\mu}sec$. Also, matured oocytes of four groups were reconstructed and fused. Reconstructed embryos were cultured in PZM-3 under 5% $CO_2$ in air at $38.5^{\circ}C$ for 6 days. The oocytes matured in the medium supplemented with FBS or/and pFF showed significantly higher maturation rates (64.0 vs. 73.9 to 85.2%). In PA embryos, cleavage rates (89.7 vs. 77.1 to 86.6%) and blastocysts rates (30.0 vs. 16.2 to 26.2%) were significantly higher in pFF group (p<0.05). In NT embryos, there was no difference among treatments in cleavage rate, but the blastocyst rates (28.5 vs. 15.5 to 24.6%) were significantly higher in pFF group (p<0.05). The apoptosis rate was significantly higher (p<0.05) in the control than other groups (10.8 vs. 4.9 to 8.2% for PA, 3.1 vs. 0.5 to 1.3% for NT). In order to select the comparable oocyte to in vivo oocytes, each group of oocytes was stained with Brilliant cresyl blue (BCB) after 42h maturation. The matured oocytes were separated according to color of cytoplasm; stained group (BCB+) and unstained group (BCB-). The oocytes matured in the presence of FBS or/and pFF showed significantly higher staining rates (70.3 to 72.7 vs. 35.1%) (p<0.05). To verify the fact that the supplementation of FBS or/and pFF can increase the maturation rates, cdc2 kinase activity, the catalytic subunit of MPF, was determined. The cdc2 kinase activity of the oocytes matured in the medium supplemented with FBS or/and pFF was significantly higher than control group (6.7 to 9.3 vs. 3.8). In conclusion, the supplementation of FBS or/and pFF can support in vitro maturation rate of porcine oocytes through the increment of cdc2 kinase activity level in the cytoplasm.

Laboratory Confirmation of A Suspicious Meningococcal Meningitis Death Case

  • Zhang Tie-Gang;He Xiong;Chen Li-Juan;He Jing-Guo;Luo Ming;Yang Jie;Shao Zhu-Jun;Sun Mei-Ping
    • Journal of Microbiology
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    • v.44 no.4
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    • pp.457-460
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    • 2006
  • A suspicious meningococcal meningitis death case was reported to the Beijing CDC. The blood specimen was analyzed via multi-PCR and MLST. 6 isolates from close contacts were analyzed via PFGE and MLST. According to the results of the above analyses, the cause of this case was identified as a serogroup A Neisseria meningitidis, which, in terms of sequence typing, belonged the ST7 group.

Effect of benzo(a)pyrene and mitomycine C on HeLa cell division cycle

  • Yu, Il-Je;Lim, Cheol-Hong;Kim, Hyo-Jung;Chung, Kyu-Hyuk;Song, Kyung-Seuk;Han, Jeong-Hee;Chung, Yong-Hyun
    • Environmental Mutagens and Carcinogens
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    • v.21 no.2
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    • pp.82-88
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    • 2001
  • Recently, there has been significant progress in understanding the control process of the cell division cycle. To investigate the influence of toxic substances on the cell cycle, the effect of benzo(a)pyrene (BAP) and mitomycine C (MMC) on synchronized HeLa cells was analyzed during the cell cycle. To synchronize the HeLa cells, 10$^{6}$ cells were grown for 1 day and then treated with 1 mM hydroxyurea for 14 h. The arrested cells were then allowed to proceed through their cell cycle by removing the hydroxyurea and resupplying a fresh medium. The arrested cells in the G1/S transition then proceeded to the S phase after 4 h, the G2/M phase after 8h, and the G1 phase after 12 h, subsequent to the resupply of a fresh medium. In the untreated HeLa cells, the p34$^{cdc2}$ kinase activity, measured using a p34$^{cdc2}$ specific peptide, peaked after 8h (G2/M) and then declined after 12 h (G1). However, treatment with 30 $\mu$M BAP delayed the peak of the p34$^{cdc2}$ kinase activity. The amount of p34$^{cdc2}$ remained unchanged in the untreated, BAP-, and MMC-treated cells throughout the cell cycle. The cyclin B level peaked after 8 h in the untreated cells, yet peaked after 10-12 h in the BAP-treated cells. There was no significant change in the cyclin B level in the MMC-treated cells.

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Molecular Mechanism of the Antiproliferative Effect by Ginseng Panaxynol on a Human Malignant Melanoma Cell Line, SK-MEL-1 (인체 흑색종 세포주 SK-MEL-1에 대한 인삼 panaxynol의 항증식 효과 기전)

  • Cho Hongkeun;Yu Su-Jin;Roh Joo Young;;Hwang Woo-Ik;Sohn Jeongwon
    • Journal of Ginseng Research
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    • v.23 no.3 s.55
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    • pp.190-197
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    • 1999
  • In this study, the molecular mechanism of the growth inhibitory effect of panaxynol was investigated in a human malignant melanoma cell line, SK-MEL-l. In the cell cycle analysis, panaxynol arrested cell cycle progression of SK-MEL-I at the G1 phase. Immunoblot analysis demonstrated that panaxynol increased $p21^{WAF1}$ and decreased cdc2 expression. Protein levels of pl6, p27, E2F-1, Rb, and p53 were not changed. Thus, the changes in expression levels of $p21^{WAF1}$ and cdc2 apparently mediate the cell cycle arrest caused by panaxynol. In addition, cycloheximide (CHX) partially reversed the growth inhibition by panaxynol, which suggested that new protein synthesis was required. On the other hand, LLnL, a proteasome inhibitor, increased antiproliferative effect of panaxynol. This may be due to stabilization of the protein(s) responsible for the growth inhibition such as $p21^{WAF1}$. In summary, these results demonstrate that panaxynol inhibits proliferation of SK-MEL-I by inducing cell cycle arrest at the G1 phase and the inhibitory effect is mediated by the increased level of $p21^{WAF1}$ as well as decreased cdc2 expression.

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Helicobacter pylori inhibited cell proliferation in human periodontal ligament fibroblasts through the Cdc25C/CDK1/cyclinB1 signaling cascade

  • Li, Huanying;Liang, Dongsheng;Hu, Naiming;Dai, Xingzhu;He, Jianing;Zhuang, Hongmin;Zhao, Wanghong
    • Journal of Periodontal and Implant Science
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    • v.49 no.3
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    • pp.138-147
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    • 2019
  • Purpose: Several studies have shown that the oral cavity is a secondary location for Helicobacter pylori colonization and that H. pylori is associated with the severity of periodontitis. This study investigated whether H. pylori had an effect on the periodontium. We established an invasion model of a standard strain of H. pylori in human periodontal ligament fibroblasts (hPDLFs), and evaluated the effects of H. pylori on cell proliferation and cell cycle progression. Methods: Different concentrations of H. pylori were used to infect hPDLFs, with 6 hours of co-culture. The multiplicity of infection in the low- and high-concentration groups was 10:1 and 100:1, respectively. The Cell Counting Kit-8 method and Ki-67 immunofluorescence were used to detect cell proliferation. Flow cytometry, quantitative real-time polymerase chain reaction, and western blots were used to detect cell cycle progression. In the high-concentration group, the invasion of H. pylori was observed by transmission electron microscopy. Results: It was found that H. pylori invaded the fibroblasts, with cytoplasmic localization. Analyses of cell proliferation and flow cytometry showed that H. pylori inhibited the proliferation of periodontal fibroblasts by causing G2 phase arrest. The inhibition of proliferation and G2 phase arrest were more obvious in the high-concentration group. In the low-concentration group, the G2 phase regulatory factors cyclin dependent kinase 1 (CDK1) and cell division cycle 25C (Cdc25C) were upregulated, while cyclin B1 was inhibited. However, in the high-concentration group, cyclin B1 was upregulated and CDK1 was inhibited. Furthermore, the deactivated states of tyrosine phosphorylation of CDK1 (CDK1-Y15) and serine phosphorylation of Cdc25C (Cdc25C-S216) were upregulated after H. pylori infection. Conclusions: In our model, H. pylori inhibited the proliferation of hPDLFs and exerted an invasive effect, causing G2 phase arrest via the Cdc25C/CDK1/cyclin B1 signaling cascade. Its inhibitory effect on proliferation was stronger in the high-concentration group.

Advanced Cooling System for CDC (일본 저온물류센터의 첨단 냉각 시스템)

  • Park, Su-Seok;Chae, Gang-Sik;Kim, Dong-Mi
    • The Magazine of the Society of Air-Conditioning and Refrigerating Engineers of Korea
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    • v.40 no.6
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    • pp.12-17
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    • 2011
  • 국내의 저온물류센터에서의 에너지 절감 방안으로서 일본의 저온 물류센터에 적용되고 있는 여러 가지 에너지 절약 시스템과 자연냉매를 이용한 친환경 시스템을 소개한다.

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Seasonal Prevalence and Integrated Control Method of Insect Pests in the Dredged Soil Dumping Area in Masan, Korea (마산시 준설토 투기장에서 발생하는 유해곤충의 계절적 발생 소장과 통합적 저감방법)

  • Kim, Yong-Ki;Lee, Cheol-Min;Bae, Yang-Seop
    • Journal of Wetlands Research
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    • v.14 no.4
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    • pp.687-698
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    • 2012
  • This study was conducted to monitor and prevent outbreak of insect pestsin dredged soil dumping area after completion of dredging construction in Masan City. Monitoring was carried out using tent trap, colored sticky trap, and CDC light trap. A total of 217,073 individuals belonging to 23 species from 10 families in 3 orders were collected. In overwintering survey using tent trap, 3 species were collected. 2 species (Leptocera fuscipennis (Haliday) and Ephydra japonica Miyagi) of them were outbreak species. In color sticky trap, more than 96% of total individuals were comprised of five species: Urolepis maritima Walker (43%), E. japonica (19%), Fucellia sp. 1 (13%), Philotelma sp. 1 (10%), and Homalometopus sp. 1 (9%). In CDC light trap, three dominant species were Homalometopus sp. 1 (91%), Glyptotendipes tokunagai Sasa (6%), and L. fuscipennis (1%), representing about 98% of the total. To prevent damage caused by outbreak of insect pests, we carried out ecological control methods such as covering the fresh soil in outbreak area, using light trap, pumping up water and so on, minimizing use of thermal fogging and insect growth regulatorwhen the insect pest population was rapidly increasing.

Long-term complications after extrahepatic cyst excision for type IV-A choledochal cysts

  • Utpal Anand;Aaron George John;Rajeev Nayan Priyadarshi;Ramesh Kumar;Basant Narayan Singh;Kunal Parasar;Bindey Kumar
    • Annals of Hepato-Biliary-Pancreatic Surgery
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    • v.27 no.4
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    • pp.433-436
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    • 2023
  • Forty-five adults with type IV-A choledochal cysts (CDC) who underwent extrahepatic cyst excision from January 2013 to December 2021 were followed up for a median interval of 25 months (range, 2 to 10 years) to observe the long-term complications in the remaining intrahepatic cyst. Late complications in varying combinations were seen in 10 patients, which included cholangitis and/or intrahepatic stones in 9 patients, intrahepatic bile duct stenosis with stones in 2 patients, anastomotic stricture in 6 patients, and left lobar atrophy with intrahepatic stones in 3 patients. Out of 6 patients who required re-do hepaticojejunostomy (HJ), three patients had left lobe atrophy with patent HJ anastomosis and a recurrent attack of cholangitis on follow-up at 3, 8, and 10 years. Complications occur frequently after extrahepatic cyst excision for type IV-A CDC and require a long-term follow-up.

Anti-cancer Effect of Hematopoietic Stem Cell-derived Allogeneic-DC Vaccine in Melanoma Metastasis Model (마우스 동종 줄기세포 유래 수지상 세포를 이용한 백신의 흑색종 폐암 전이 모델에서의 항암 효과 및 기전 연구)

  • Kim, Myoung-Joo;Shon, Hye-Jin;Baek, So-Young;Lee, Kang-Eun;Lee, Young-Joon;Lee, Hyun-Ah
    • IMMUNE NETWORK
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    • v.6 no.3
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    • pp.154-162
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    • 2006
  • Background: Dendritic cell (DC)-based cancer immunotherapy is studied for several years. However, it is mainly derived from autologous PBMC or leukapheresis from patient, which has limitations about yield and ability of DC production according to individual status. In order to solve these problems, inquiries about allogeneic DCs are performed but there are no preclinical trial answers for effect or toxicity of allogeneic DC to use for clinical trial. In this study, we compared the anti-tumor effect of allogeneic and autologous DCs from mouse bone marrow stem cells in mouse metastatic melanoma model. Methods: B16F10 melanoma cells ($5{\times}10^4$/mouse) were injected intravenously into the C57BL/6 mouse. Therapeutic DCs were differentiated from autologous (C57BL/6: CDC) or allogeneic (B6C3F1: BDC) bone marrow stem cells with GM-CSF, SCF and IL-4 for 13days and pulsed with B16F10 tumor cell lysate (Blys) for 18hrs. DC intra-peritoneal injections began on the 8th day after the tumor cell injection by twice with one week interval. Results: Anti-tumor response was observed by DC treatment without any toxicity especially in allogeneic DC treated mice (tumor burden score: $2.667{\pm}0.184,\;2.500{\pm}0.463,\;2.000{\pm}0.286,\;1.500{\pm}0.286,\;1.667 {\pm}0.297$ for saline, CDC/unpulsed-DC: U-DC, CDC/Blys-DC, BDC/U-DC and BDC/Blys-DC, respectively). IFN-${\gamma}$ secretion was significantly increased in allogeneic DC group stimulated with B16F10 cell lysate ($2,643.3{\pm}5,89.7,\;8,561.5{\pm}2,204.9.\;6,901.2{\pm}141.1pg/1{\times}10^6$ cells for saline, BDC/U-DC and BDC/Blys-DC, respectively) with increased NK cell activity. Conclusion: Conclusively, promising data was obtained that allogeneic DC can be used for DC-based cancer immunotherapy.