• Title/Summary/Keyword: Surface labelling

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Parthenogenetic Mouse Embryonic Stem Cells have Similar Characteristics to In Vitro Fertilization mES Cells (체외수정 유래 생쥐 배아줄기세포와 유사한 특성을 보유한 단위발생 유래 생쥐 배아줄기세포)

  • Park, Se-Pill;Kim, Eun-Young;Lee, Keum-Si;Lee, Young-Jae;Shin, Hyun-Ah;Min, Hyun-Jung;Lee, Hoon-Taek;Chung, Kil-Saeng;Lim, Jin-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.29 no.2
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    • pp.129-138
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    • 2002
  • Objective: This study was to compare the characteristics between parthenogenetic mES (P-mES) cells and in vitro fertilization mES cells. Materials and Methods: Mouse oocytes were recovered from superovulated 4 wks hybrid F1 (C57BL/6xCBA/N) female mice. For parthenogenetic activation, oocytes were treated with 7% ethanol for 5 min and $5{\mu}g$/ml cytochalasin-B for 4 h. For IVF, oocytes were inseminated with epididymal sperm of hybrid F1 male mice ($1{times}10^6/ml$). IVF and parthenogenetic embryos were cultured in M16 medium for 4 days. Cell number count of blastocysts in those two groups was taken by differential labelling using propidium iodide (red) and bisbenzimide (blue). To establish ES cells, b1astocysts in IVF and parthenogenetic groups were treated by immunosurgery and recovered inner cell mass (ICM) cells were cultured in LIF added ES culture medium. To identify ES cells, the surface markers alkaline phosphatase, SSEA-1, 3,4 and Oct4 staining were examined in rep1ated ICM colonies. Chromosome numbers in P-mES and mES were checked. Also, in vitro differentiation potential of P-mES and mES was examined. Results: Although the cleavage rate (${\geq}$2-cell) was not different between IVF (76.3%) and parthenogenetic group (67.0%), in vitro development rate was significantly low in parthenogenetic group (24.0%) than IVF group (68.4%) (p<0.05). Cell number count of ICM and total cell in parthenogenetic b1astocysts ($9.6{\pm}3.1,\;35.1{\pm}5.2$) were signficantly lower than those of IVF blastocysts ($19.5{\pm}4.7,\;63.2{\pm}13.0$) (p<0.05). Through the serial treatment procedure such as immunosurgery, plating of ICM and colony formation, two ICM colonies in IVF group (mES, 10.0%) and three ICM colonies (P-mES, 42.9%) in parthenogenetic group were able to culture for extended duration (25 and 20 passages, respectively). Using surface markers, alkaline phosphatase, SSEA-l and Oct4 in P-mES and mES colony were positively stained. The number of chromosome was normal in ES colony from two groups. Also, in vitro neural and cardiac cell differentiation derived from mES or P-mES cells was confirmed. Conclusion: This study suggested that P-mES cells can be successfully established and that those cell lines have similar characteristics to mES cells.

"I'm healthy, I don't have pain"- health screening participation and its association with chronic pain in a low socioeconomic status Singaporean population

  • Wee, Liang En;Sin, David;Cher, Wen Qi;Li, Zong Chen;Tsang, Tammy;Shibli, Sabina;Koh, Gerald
    • The Korean Journal of Pain
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    • v.30 no.1
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    • pp.34-43
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    • 2017
  • Background: We sought to determine the association between chronic pain and participating in routine health screening in a low socioeconomic-status (SES) rental-flat community in Singapore. In Singapore, ${\geq}85%$ own homes; public rental flats are reserved for those with low-income. Methods: Chronic pain was defined as pain ${\geq}3$ months. From 2009-2014, residents aged 40-60 years in five public rental-flat enclaves were surveyed for chronic pain; participation in health screening was also measured. We compared them to residents staying in adjacent owner-occupied public housing. We also conducted a qualitative study to better understand the relationship between chronic pain and health screening participation amongst residents in these low-SES enclaves. Results: In the rental-flat population, chronic pain was associated with higher participation in screening for diabetes (aOR = 2.11, CI = 1.36-3.27, P < 0.001), dyslipidemia (aOR = 2.06, CI = 1.25-3.39, P = 0.005), colorectal cancer (aOR = 2.28, CI = 1.18-4.40, P = 0.014), cervical cancer (aOR = 2.65, CI = 1.34-5.23, P = 0.005) and breast cancer (aOR = 3.52, CI = 1.94-6.41, P < 0.001); this association was not present in the owner-occupied population. Three main themes emerged from our qualitative analysis of the link between chronic pain and screening participation: pain as an association of "major illness"; screening as a search for answers to pain; and labelling pain as an end in itself. Conclusions: Chronic pain was associated with higher cardiovascular and cancer screening participation in the low-SES population. In low-SES populations with limited access to pain management services, chronic pain issues may surface during routine health screening.

INFLUENCE OF TOPICAL IRRIGATION USING THE HA & PURE Ti IMPLANTS ON BONE FORMATION;A STUDY ON THE IRRADIATED RABBIT TIBIA (방사선 조사후 매식한 임프란트와 관주에 따른 골치유에 대한 비교연구)

  • Hong, Sung-Pal;Rhee, Gun-Joo;Cha, Yong-Doo;Oh, Se-Jong;Hyun, Jung-Min;Choi, Dong-Joo;Park, Young-Joo;Park, Jon-Woo
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.26 no.1
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    • pp.59-72
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    • 2000
  • In this study, the rate of bone formation and the pattern of bone to implant contact surface around HA coated implant and pure Ti implant inserted into the irradiated tibia of rabbit were compared. Sixteen mongrel mature male rabbits were used as experimental animal. Each rabbit received 15 Gy of irradiation. Four weeks after irradiation, two holes were prepared on the tibia of each rabbit for placement of HA coated type and pure Ti type implants. Prior to implant placement, one group received steroid irrigation and the control group was similarly irrigated with normal saline. This was immediately followed by placement of the two different types of implants. Postoperatively, tetracycline was injected intramuscularly for 3 days. For fluorescent labelling, 3 days of intramuscular alizarine red injection was given. 2 weeks before sacrifice, followed by intramuscular calcein green on the last 3 days before specimen collection. Each rabbit was sacrificed on the second, fourth, sixth and eighth week after the implantation. The specimens were observed by the light microscope and the fluorescent microscope. The results were as follows; 1. All implants inserted into the irradiated tibia of rabbit were free from clinical mobility and no signs of bony resorption were noted around the site of implant placement. 2. Under the light microscope, new bone formation proceeded faster around implants that received steroid irrigation compared to the control group irrigated with saline. Bone to implant contact surface was greater in the steroid irrigated group than the saline irrigated group. Therefore, better initial stabilization was observed in the group pretreated with steroid irrigation. 3. Under the light microscope. HA coated implants showed broader bone to implant contact surface than pure Ti implants, and HA coated implants had better bone healing pattern than pure Ti implants. 4. In the steroid pretreated group, acceleration of bone formation was demonstrated by fluorescent microscopy around the 2, 4 weeks group and the 6 weeks HA coated implant group. The difference in the rate of bone formation proved to be statistically significant(P<0.05). Faster bone formation was noted in the saline irrigated group in the 6 weeks pure Ti implants and 8 weeks group. The difference was not statistically significant(P<0.05). 5. For the rabbits that were sacrificed on the second and fourth week after the implant placements, the rates of bone formation around HA coated implants proceeded faster than those around pure Ti implants under the fluorescent microscopy. For the rabbits that were sacrificed on the sixth week after the implant placements, the rates of bone formation around pure Ti implants proceeded faster than those around HA coated implants under the fluorescent microscopy. But this result did not show statistical significance (P<0.05) For the rabbits that were sacrificed on the eighth week after the implant placements, the rates of bone formation around HA coated implants proceeded faster than those around pure Ti implants under the fluorescent microscopy. This result was statistically significant (P<0.05).

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