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| 1 | Introduction of DT40 cells into chick embryos显示文摘To examine the transfection of exogenous genes into chick embryos, applying the characteristics of avian leukosis virus (ALV)-induced chicken B cell line DT40 to the production of chimeric birds. Methods: The DT40cells incorporated with exogenous gene (lacZ constructs encoding Escherichia coli β-galactosidase: β-gal) were introduced into chick embryos by the injection of cells into stage X blastoderm. Manipulated eggs were incubated for 3 (trial 1 ) or 6 (trial 2) days, and the expression of lacZ DNA was detected by a histochemical staining method of β-galactosidase and polymerase chain reaction (PCR) analysis. Results: The survival rates of the manipulated embryos incubated for 3 days (stage 18-20: trial 1) and 6 days (stage 28, 30: trial 2) were about 42% and 38%, respectively.The expression rates of the lacZ gene in the embryos in the trials 1 and 2 were about 60% and 23%, respectively, for the survived embryos. Conclusio: The rate of embryonic viability and expression rate of introduced genes were not so high, but it suggested the possibility of utilizing the DT40 cells as a vector for carrying exogenous genes into chick embryos. | Mariko Toba Fumio Ebara Hiroki Furuta Yuichi Matsushimal Yasuo Kitagawa Noboru Fujihara | 2001 | Asian Journal of Andrology2001，3，1： | 3 |
| 2 | Successful closing of duodenal ulcer after endoscopic submucosal dissection with over‐the‐scope clip to prevent delayed perforation显示文摘 | Hirohito Mori Fujihara Shintaro Hideki Kobara Noriko Nishiyama Kazi Rafiq Mitsuyoshi Kobayashi Toshiaki Nakatsu Noboru Miichi Yasuyuki Suzuki Tsutomu Masaki | 2013 | Digestive Endoscopy2013，，4： | 3 |
| 3 | 家兔细胞核移植技术研究显示文摘以家兔 1 6~ 3 2细胞期胚胎卵裂球为核供体 ,显微注射到去核卵母细胞卵周隙中 ,共制作完成 2 2 9枚重组胚 ,2次电融合后融合率达到 76.8%。重组胚与兔胚胎成纤维细胞共同培养 ,卵裂率为 65 .7% (2 3 /3 5 ) ,桑囊率达到 2 2 .9% (8/ 3 5 ) ,均显著高于 (P<0 .0 1 )微滴直接培养结果 (4 3 .3 % ,1 3 / 3 0 ;1 6.7% ,5 / 3 0 )。将1 1 1枚重组胚手术移植到 9只同期发情的受体母兔输卵管中 ,2只妊娠 ,其中 1只完成足月胚胎发育产出克隆仔兔 2只。 | 常万存 韩建永 马世援 张红卫 陈忠科 Noboru Fujihara 王庆国 刘文浩 | 2002 | 西北农业学报2002，11，1： | 2 |
| 4 | Evaluation of deoxyribonuclease activity in seminal plasmaof ejaculated chicken semen显示文摘Aim: To confirm the stability of exogenous genes in the generation of transgenic chickens using ejaculated chicken sperm, the deoxyribonuclease (DNase) activity was evaluated in the seminal plasma of ejaculated semen and the stability of DNA was examined by adding lipofection reagents. Methods: A PCR fragment (249 bp) of pEGFPN-1 vector was used as the DNA substrate and was incubated with the seminal plasma at 40℃for 30 min. Then, the whole reaction solution was subjected to agarose gel electrophoresis and the DNA size was evaluated under UV light.Results: The DNA substrate was completely diminished after incubation with seminal plasma. However, the sub-strate was intact after incubation with heat-treated seminal plasma or incubation with seminal plasma in the presence of 0.5 mmol/L-5 mmol/L EDTA. The substrate was stabilized in the seminal plasma by the addition of commercially available lipofection reagents. Conclusion: The DNase activity is present in the seminal plasma of ejaculated chicken semen. However, DNA is stable in the liposomal-DNA complex. ( Asian JAndro12003 Sep; 5: 213-216) | Fuminori Sato Tomoki Soh Masa-aki Hattori Noboru Fujihara | 2003 | Asian Journal of Andrology2003，5，3： | 1 |
| 5 | Reproductive characteristics of transgenic (TG) chickens caping an exogenous gene显示文摘An exogenous gene (lacZ/MiwZ) introduced into the germinal crescent region (GCR) of avian embryos was con-finned to be successfully transferred to the gonads via the primordial germ ceils (PGCs). Following hatching, the chinkswere raised until the stage of sexual maturation. The incorporation of MiwZ DNA was detected in male and female trans-genic chickens, respectively. The normal male and female transgenic birds were subjected to artificial insemfution according to routine methods. Fertilized eggs obtained from female transgenic chickens were incubated for 7'2 h and the em-bryos removed from the yolk were examined by X-gal staining to detect the introduction of MiwZ in the offspring. As a result, | Furmio Ebara Noboru Fujihara | 1999 | Asian Journal of Andrology1999，1，3： | 0 |
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