• Clinical and Experimental Reproductive Medicine
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• v.19 no.2
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• pp.143-152
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• 1992

The present experiments were focussed to modify a short slow-cooling protocol used for freezing of early stage embryo(Testart et al., 1986) and also to apply the modified method for the cryopreservation of hamster oocytes with Zona or without. The protocol was modified by changing the 4-step equilibration into 1-step and the 1-step thawing into 2-step. The oocytes were added in 1.5M PROH and 0.1M Sucrose, seeded at $-7^{\circ}C$, slow cooled($0.3^{\circ}C$/min) to $-30^{\circ}C$ before plunging to $-196^{\circ}C$. The oocytes were thawed at $23-25^{\circ}C$ air(20sec/150sec) and/or 33-35 water(10sec). The survival of the frozen-thawed oocytes was determined by morphologic criteria and their fertilizing ability was also estimated by Sperm Penetration Assay(SPA) system(Chang et al, 1990) using fertile men semen sample. One-step equilibration showed slightly higher survival rate(83.9% vs. 71.0%) and fertilization rate(83.9% vs. 71.0%) compared with four-step(p>0.05). And two-step thawing(air & water exposing) of oocytes frozen after 1-step equilibration showed significantly higher survival rate(96.3%) than one-step thawing at air(85.2%) or water(65.0%) only(p<0.05). Therefore, by the modified method(l-step equilibration & 2-step thawing), Zona-intact(ZI) and Zona-free(ZF) oocytes were frozen and thawed. ZI-oocytes showed significantly higher survival rate(95.4%, 308/323 vs. 67.6%, 240/355) than ZF-oocytes(P<0.01). But the survival of ZF-oocytes was as high as ZI-oocytes in fourteen of twenty-four replicates. ZI-oocytes was also significantly higher fertilization rate($92.4{\pm}8.9%$ vs. $63.7{\pm}18.5%$) and higher mean number of penetrated sperm($6.2{\pm}4.2$ vs. $3.9{\\pm}3.3$) than ZF-oocytes, but not higher than control(fresh oocytes;$99.3{\pm}2.4%$, $8.4{\pm}4.2$)(P<0.001). Conclusively, this modified method will contribute to freeze effectively the hamster oocytes for simplifing of the logical consideration of performing SPA and also to freeze the human and other animal oocytes.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.1
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• pp.23-28
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• 2003

The present study was undertaken to investigate the effects of PVP concentration and exposure temperature to vitrification solution on the post-thaw survival, in vitro maturation and development of immature bovine oocytes (germinal vesicle stage). The vitrification solution (VS) consisted of 40% ethylene glycol (EG)+0.5 M sucrose (S)+10% FBS. PVP was added to VS: 0%, 5% or 10%. The cumulus-oocyte complexes (COCs) were diluted in VS as one step, after 2 min the COCs were loaded in straw and vitrified by direct immersion into liquid nitrogen. For thawing, the straws were plunged into $30^{\circ}C$ water bath for 10s. After thawing, the oocytes were diluted in 0.5 M (in DPBS with 10% FBS) sucrose solution for 5 min. The survival rate (FDA-test and trypan blue) of immature bovine oocytes was measured. The survival rate was higher in 5% PVP (91.5%) than in 0% (64.2%) or in 10% PVP (79.7%). The proportion of metaphase II formation was 69.35% in control (no vitrified COCs), 9.3% in 40% EG+0.5 M S+0% PVP and 21.05% in 40% EG+0.5 M S+5% PVP (p<0.05). The effect of room temperature ($25^{\circ}C$ for 10 min) and cold temperature ($4^{\circ}C$ for 10 min) on COCs were determined in this study. After IVF, the cleavage and blastocysts rate of oocytes exposed to room temperature and cold temperature in VS+5% PVP was significantly different (2 cell: 63.20% vs 37.97%, blastocysts: 18.40% vs 2.53%). The cleavage rates of frozen-thawed oocytes were 20.53% with PVP and 22.13% without PVP (p>0.05). Two out of 151 oocytes (1.32%) developed to blastocyst stage after frozen-thawed with 5% PVP (p>0.05). Development of oocytes after frozen-thawing to the 2 cell were not significantly affected with or without PVP following IVF. However, the vitrification of immature bovine oocytes with PVP maintained the ability to develop to the blastocyst stage after IVM-IVF and IVC, while no blastocysts were obtained from oocytes vitrified without PVP. These results suggested that PVP has a protective role for vitrification of immature bovine oocytes as far as survival is concerned, however, the protection was not sufficient enough to support blastocyst formation.

• Journal of Korean Society of Forest Science
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• v.40 no.1
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• pp.43-50
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• 1978

The elder berry was known to the rich in natural food colour resources and used in as for making wine, confectionary, perfumes, natural food colour, making elder berry juice, jelly, jam and medicinal properties or oils. In the present study, wish was to find the effect of various factors on the success of the vegetative propagation of elder berry by means cutting in the exposed field and green house was carried out and those obtained results can be summarized as follows. 1. Cuttings with dormant cutting stocs in the polyethylen house with heating and water mist spray resulting 100 percent survival. And temperature and the relative humidity in which fraim during the cutting season were around $20{\sim}25^{\circ}C$ and 70-90% respectively and in case as more significant other of the 1% Level. 2. With five varieties tested, resulting 93.8 percent survival, The F. value is not significant. 3. With four organs of cutting stock tested resulting 57.5 percent survival on the cuttings with two knodes of dormant cutting stock served as better cutting stock than others. The F. value is more significantly 1% Level. 4. Dormand bud served as possible cutting stock was found to be 17.66 survival percentage. 5. Both earlier and later stage of germinated Leaves with soft wood cutting stock poor cuttings, and the degree of development of 15th June cutting stock was optimum stage on the principal factor governing the success of cutting in the soft wood cutting showing 54% survival. The F. value is more 1% Level significant.

• Journal of Veterinary Science
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• v.23 no.1
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• pp.8.1-8.15
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• 2022

Background: Brucella infection induces brucellosis, a zoonotic disease. The intracellular circulation process and virulence of Brucella mainly depend on its type IV secretion system (T4SS) expressing secretory effectors. Secreted protein BspJ is a nucleomodulin of Brucella that invades the host cell nucleus. BspJ mediates host energy synthesis and apoptosis through interaction with proteins. However, the mechanism of BspJ as it affects the intracellular survival of Brucella remains to be clarified. Objectives: To verify the functions of nucleomodulin BspJ in Brucella's intracellular infection cycles. Methods: Constructed Brucella abortus BspJ gene deletion strain (B. abortus ∆BspJ) and complement strain (B. abortus pBspJ) and studied their roles in the proliferation of Brucella both in vivo and in vitro. Results: BspJ gene deletion reduced the survival and intracellular proliferation of Brucella at the replicating Brucella-containing vacuoles (rBCV) stage. Compared with the parent strain, the colonization ability of the bacteria in mice was significantly reduced, causing less inflammatory infiltration and pathological damage. We also found that the knockout of BspJ altered the secretion of cytokines (interleukin [IL]-6, IL-1β, IL-10, tumor necrosis factor-α, interferon-γ) in host cells and in mice to affect the intracellular survival of Brucella. Conclusions: BspJ is extremely important for the circulatory proliferation of Brucella in the host, and it may be involved in a previously unknown mechanism of Brucella's intracellular survival.

• Asian Journal of Innovation and Policy
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• v.6 no.3
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• pp.274-294
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• 2017

While studies have viewed the effect of Chinese talent-attracting programs launched by government since reform and open door policy, little of them has assessed these programs empirically and pertinently. This article intends to assess an important program - the Thousand Youth Talents Program (TYTP). Frist, this paper proposed a transnational migration matrix of the academics to clarify the dynamic mechanism of academic brain gain at the high end. Then, the Kaplan-Meier analysis and Cox regression model are used to empirically analyze the policy effect of TYTP. The results show that, academic ability have double edged impacts on brain gain at the high end, some scholars whose last employer's academic ranking is world's Top100 have stronger willing to return, and the negative effect of academic ranking decreases with time passing; while scholars with a tenure-track position, a tenure position or a permanent position tend to stay overseas, and the hazard rate of staying increases with age. The older scholars have more intentions to go back China, while gender was not a significant factor influencing academic return at the high end. That is, the talent-attracting programs has partly succeeded in bringing back the academics at the high end.

• International Journal of Industrial Entomology and Biomaterials
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• v.13 no.2
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• pp.63-72
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• 2006

Parthenogenesis, the development of unfertilized ovum opens new perspectives in silkworm breeding in the development of homozygous breeds. In order to improve induction of artificial parthenogenesis in the excised unfertilized eggs of different breeds of the silkworm, Bombyx mori L., a new method was devised and the results were compared with the routine method. General and specific combining abilities and hybrid vigour of newly developed bivoltine breeds were analyzed utilizing bivoltine breeds viz., $CSR_2,\;CSR_4,\;CSR_{17}\;and\;NB_4D_2$ Estimation of GCA revealed superiority of the breeds, $DNB_1$ for eight characters followed by $DNB_4$ for five characters. Among the testers, $CSR_2$ was found good general combiner for seven characters followed by $CSR_{17}$ for four characters. A great deal of variations was observed among the hybrids studied. Five hybrids namely, $DNB_1{\times}CSR_2,\;DNB_4{\times}CSR_4,\;DNB_4{\times}NB_4D_2,\;DNB_6{\times}CSR_2\;and\;DNB_7{\times}CSR_2$ showed significant SCA effects for 5-6 characters. The hybrid, $DNB_4{\times}CSR_4$ showed its superiority by expressing significant hybrid vigour over BPV for 7 characters. Majority of the hybrids exhibited significant hybrid vigour for survival rate, yield/10,000 larvae by weight, cocoon weight, cocoon shell weight, filament length and denier.

• IMMUNE NETWORK
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• v.13 no.1
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• pp.16-24
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• 2013

CTLA-4Ig is regarded as an inhibitory agent of the T cell proliferation via blocking the costimulatory signal which is essential for full T cell activation. To improve applicability, we developed the CTLA-4Ig-CTKC in which the c-terminal lysine had been replaced by cysteine through single amino acid change. The single amino acid mutation of c-terminus of CTLA-4Ig was performed by PCR and was checked by in vitro transcription and translation. DNA construct of mutant form was transfected to Chinese hamster ovary (CHO) cells by electroporation. The purified proteins were confirmed by Western blot and B7-1 binding assay for their binding ability. The suppressive capacity of CTLA-4Ig-CTKC was evaluated by the mixed lymphocyte reaction (MLR) and in the allogeneic pancreatic islet transplantation model. CTLA-4Ig-CTKC maintained binding ability to B7-1 molecule and effectively inhibits T cell proliferation in MLR. In the murine allogeneic pancreatic islet transplantation, short-term treatment of CTLA-4Ig-CTKC prolonged the graft survival over 100 days. CTLA-4Ig-CTKC effectively inhibits immune response both in MLR and in allogeneic islet transplantation model, indicating that single amino acid mutation does not affect the inhibitory function of CTLA-4Ig. CTLA-4Ig-CTKC can be used in vehicle-mediated drug delivery system such as liposome conjugation.

• Journal of Embryo Transfer
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• v.21 no.3
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• pp.255-262
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• 2006

The purpose of this study is to investigate the effects of vitrification in open pulled straws (OPS) methods on in vitro survival ability of porcine embryos. For in vitro maturation of immature oocytes, the porcine ovaries were collected from local slaughter-house. The cumulus-oocytes complexes were aspirated from 2 to 6 mm follicles. The collected oocytes were cultured for in vitro maturation in NCSU-23 medium with 5 mM hypotaurine, 0.57 mM cysteine, 10% porcine follicle fluid, 10 IU/ml PMSG and 10 IU/ml hCG for $21{\sim}22$ hrs. Then, the oocytes were more cultured $21{\sim}22$ hrs in vitro maturation in medium removed hormones. The frozen-thawed spermatozoa were washed by centrifugation 2 times for 10 min at 1,500 rpm in D-PBS with 5.56 mM glucose, 0.33 mM Na-pyruvate, 100 IU/ml penicillin, $100 {\mu}g/ml$ streptomycin and 4 mg/ml BSA. The fertilization medium used mTBM with 2 mM caffeine and 2 mg/ml BSA and adjusted to a pH of 7.2 to 7.4. The final concentration of spermatozoa was adjusted to $2.5{\times}10^6$cells/ml motile sperm during fertilization in vitro. At 8 hrs after insemination, the oocytes were transferred into NCSU-23 medium with 5.0 mM hypotaurine, 4 mg/ml BSA and 10 ng/ml EGF and cultured for 7 days. When the blastocysts of different stages were frozen-thawed by OPS methods, the proportions of embryos with normal morphology were significantly (p<0.05) higher in embryos frozen-thawed at expanded blastocyst stage (38.9%) than in early blastocyst stage (28.3%). On the other hand, the proportions of embryos damaged after frozen-thawing were significantly (p<0.05) higher in embryos frozen at early blastocyst stages than in expanded blastocyst stage. In another experiment, the normal embryos morphology after frozen- thawing were further cultured for 48 hrs. After culture, the proportions of embryos hatched were 6.7, 20.0 and 33.3% for embryos frozen-thawed at early blastocyst, mid-blastocyst and expanded blastocyst stages. These finding indicate the possible broader application for OPS methods, as frozen-thawed embryos may be accompanied by developmental stage according to requirements of the survival ability after freezing of blastocyst stage in the pig.

• Korean Journal of Poultry Science
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• v.48 no.2
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• pp.69-80
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• 2021

We conducted a 4 × 4 diallel cross-mating test using 934 chickens from four grandparent stock lines to develop a new synthetic breed of Korean native chicken. The mean values, combining ability, and reciprocal effects on survival rate, body weight, and hen-day egg production were analyzed. In phenotypes, most chickens have yellowish-brown, reddish-brown and mixed color feathers. The average survival rate was 86.8±12.3%, with the highest in YH combination. Specific combining ability (SCA) had a greater effect on survival rate than general combining ability (GCA), and the SCA of HY combination was the highest. The 16 cross-combinations were distinctly divided into three weight groups according to their genetic characteristics. At 12 weeks of age, GCA showed a greater effect on weight than SCA, and the SCA of FH combination was the highest. The age at first egg laying was 157 days, and the crosses reached sexual maturity faster than the pure lines. The egg production rate was highest in SY at 79.5±2.1%. The GCA and SCA for hen-day egg production were similar, and the SCA was highest in the HS and FY combinations. The reciprocal effect showed that the offspring's egg production rate was high when S and Y were maternal parents in almost all combinations. In conclusion, FH and HF combinations, which have excellent growth performance, are the most desirable paternal parent stock strains, and FY, FS, HY, and SY combinations, which have excellent laying performance with moderate weight, are the preferred maternal strains.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.53 no.2
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• pp.191-202
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• 2020

We evaluated the effects of dietary supplementation of phytosterol on the growth, feed utilization, immunity, digestibility, wound healing ability and disease resistance of olive flounder Paralichthys olivaceus. We conducted two consecutive feeding trials at different growth stages of the fish: EXP-1 (68.9 g) and EXP-2 (16.5 g). The experimental diets were supplemented with graded levels of phytosterol (1% ecdysteroid extracted from Achyranthis radix) at 0, 0.025, 0.05, 0.1, 0.2 and 0.4% (designated as Con, PHY0.025, PHY0.05, PHY0.1, PHY0.2 and PHY0.4, respectively). Dietary phytosterol did not significantly affect growth and cholesterol concentration. Feed utilization was higher in fish fed phytosterol-supplemented diets than in fish fed the control diet. Dietary phytosterol increased innate immunity and digestibility of protein and dry matter. Wound healing ability was also increased by the phytosterol supplementation. The survival against Edwardsiella tarda challenge was higher in fish fed low-phytosterol diets than in fish fed the control diet. The optimum dietary level of phytosterol seems to be approximately 0.05%. The results in this study indicate that the phytosterol could be used as a functional supplement in diets to improve feed utilization, immunity, digestibility and wound healing ability of olive flounder.