• 제목/요약/키워드: CDC6

검색결과 125건 처리시간 0.026초

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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    • 제33권2호
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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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    • 제44권4호
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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
    • 한국환경성돌연변이발암원학회지
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    • 제21권2호
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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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인체 흑색종 세포주 SK-MEL-1에 대한 인삼 panaxynol의 항증식 효과 기전 (Molecular Mechanism of the Antiproliferative Effect by Ginseng Panaxynol on a Human Malignant Melanoma Cell Line, SK-MEL-1)

  • 조홍근;유수진;노주영;하영미;황우익;손정원
    • Journal of Ginseng Research
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    • 제23권3호
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    • pp.190-197
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    • 1999
  • 본 연구는 인삼 panaxynol이 인체 흑색종 세포주 SK-MEL-1 미치는 항증식효과의 분자적 기전을 알아보고자, 세포주기와 세포주기 조절인자들의 발현변화, 단백질 합성 억제제와 proteasome억제제가 panaxynol의 항암효과에 미치는 영향을 조사하였다. panaxynol은 세포주기의 G1 단계 진행을 억제시켰으며, 동시에 $p21^{WAF1}$ 발현 증가와 cdc2의 발현 감소를 유발하였다. 이에 비해 p16, p27, E2F-1, Rb, p53의 발현에는 변화가 없었다. 이 결과는 panaxynol의 SK-MEL-1 세포에서 $p21^{WAF1}$의 발현을 증가시키고 cdc2의 발현을 감소시켜, 세포주기의 G1-S 이행 단계를 억제한다는 것을 보여준다 또한 CHX는 panaxynol의 항암효과를 감소시키고, LLnL은 panaxynol의 항암 효과를 증가시켰는데, 이는 panaxynol에 의한 SK-MEL-1 세포의 증식억제에 새로운 단백질 합성이 필요하며, LLnL이 panaxyno떼 의한 세포증식억제를 매개하는 $p21^{WAF1}$등의 단백질 분해를 저해시키기 때문인 것으로 생각된다.

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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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    • 제49권3호
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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.

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

  • 김용기;이철민;배양섭
    • 한국습지학회지
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    • 제14권4호
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    • pp.687-698
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    • 2012
  • 본 연구는 마산시에서 준설공사 완료 후 준설토 투기장에서 유해곤충의 대량발생을 조사하고, 피해를 예방하기위해서 실시되었다. 모니터링은 텐트트랩, 색상착상트랩, CDC라이트 트랩을 이용하여 실시하였다. 조사결과, 3목 10과 23종 217,073개체의 곤충류가 채집되었다. 텐트트랩을 이용한 월동조사에서 채집된 3종 중 2종(애기똥파리(Leptocera fuscipennis (Haliday))와 극동물가파리(Ephydra japonica Miyagi)는 대량발생 종이었다. 색상점착트랩에서는 16종 중 5종(물가파리금좀벌(Urolepis maritima Walker) (43%), 극동물가파리(19%), Fucellia sp. 1(13%), Philotelma sp. 1(10%), Homalometopus sp. 1(9%))이 전체 개체수의 약 96%를 차지하였다. CDC 라이트트랩에서 우점종은 Homalometopus sp. 1(91%), 도꾸나가조각깔따구(Glyptotendipes tokunagai Sasa) (6%), 애기똥파리(1%)로 전체의 약 98%를 차지했다. 조사시 개체수의 급격한 증가가 관찰되었을 경우에는 유해곤충의 대량발생으로 인한 피해를 막기 위해서, 가열연막과 유충성장억제제 사용을 최소화하면서, 대량발생지역 복토, 유문등 사용, 물빼기작업 등의 환경적인 저감방법을 수행하여 그 결과를 분석하였다.

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
    • 한국간담췌외과학회지
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    • 제27권4호
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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.

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

  • 김명주;손혜진;백소영;이강은;이영준;이현아
    • IMMUNE NETWORK
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    • 제6권3호
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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.