• Reproductive and Developmental Biology
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• 제30권2호
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• pp.81-85
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• 2006

This study was conducted to examine the effect of IRES controlled reporter gene on screening and production of recombinant human erythropoietin (EPO) proteins from cultured CHO cells. The cDNA was cloned for EPO from human liver cDNA Using site-directed mutagenesis, we generated recombinant human EPO (rhEPO) with two additional N-glycosylations (Novel erythropoiesis-stimulating protein: NESP). Wild type hEPO and NESP were cloned into expression vectors with GFP reporter gene under regulatory control of CMV promoter and IRES so that the vectors could express both rhEPO and GFP. The expression vectors were transfected to cultured CHO-K1 cells. Under microscopy, expression of GFP was visible. Using supernatant of the culture, ELISA assay, immunocytochemistry and in vitro assay using EPO dependant cell line were performed to estimate biological activity to compare the production characteristics (secretion levels, etc.) between rhEPO and NESP. The activity of NESP protein, obtained by mutagenesis, was described and compared with its rhEPO counterpart produced under same conditions. Although NESP had less secretion level in CHO cell line, the biological activity of NESP was greater than that of rhEPO. These results are consistent with previous researches. We also demonstrated that rhEPO and GFP proteins expressed simultaneously from transfected CHO cell line. Therefore we conclude that use of GFP reporter gene under IRES control could be used to screen and produce rhEPO in cultured CHO cells.

• 혜화의학회지
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• 제16권1호
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• pp.41-59
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• 2007

From the study of prescription on low back pain, the following conclusions are obtained. 1. Among classified cause of low back pain, ShinHur(腎虛) lowback pain and its prescription was most mentioned. 2. Prescriptions such as ChungAWhan(靑娥丸), KookBangAnShinWhan(局方安腎丸) BoSooDan(補髓丹) BaekBaeWhan(百倍丸) DooChungWhan(杜沖丸) JangBonDan(壯本丹) NokKakWhan(鹿角丸) were used in ShinHur(腎虛) type low back pain. 3. Prescription such as TaekRanTang(澤蘭湯) JiRyongSan(地龍散) YoeShinSan(如神散) ShinKookJoo(神麴酒) SoeGuenSan(舒筋散) were used in JwaSumJilBak(閃挫跌撲) type low back pain. 4. Prescription such as ChangChulTang(蒼朮湯) JumTongTang(拈痛湯) ChulBuTang(朮附湯) YiChoChangBaekSan(二炒蒼栢散) were used in SeupYoel(濕熱) type low back pain. 5. Prescription such as ChunGoongYookGaeTang(川芎肉桂湯) GaMiSaMulTang(加味四物湯) PaHoelSanDongTang(破血散疼湯) JiRyongSan(地龍散) were used in UhHoel(瘀血) type low back pain. 6. Prescription such as (蒼術復煎散) (五積散) (摩腰丹) (滲濕湯) were used in HanSeup(寒濕) type low back pain. 7. Prescription such as GaMiYiJinTang(加味二陳湯) GongYeonDan(控涎丹) SaMoolTangHapYiJinTangGaMi(四物湯合二陳湯加味) were used in DamUem(痰飮) type low back pain. 8. Prescription such as OhJukSanGaMi(五積散加味) OhYakSoonGiSanGaMi(烏藥順氣散加味) GaMiYongHoSan(加味龍虎散) SoSokMyoungTang(小續命湯) were used in Poong(風) type low back pain. 9. Prescription such as (四物湯合二陳湯) (仰腰湯) were used in SikJuk(食積) type low back pain and (五積散) (煨腎散) (三花神祐丸) in Seup(濕) type low back pain. 10. Prescription such as ChilKiTang(七氣湯) ChimHyangGangKiTang(沈香降氣湯) ChoKiSan(調氣散) InSamSoonKiSan(人參順氣散) were used in Ki(氣) type low back pain.

• 한국수정란이식학회지
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• 제25권3호
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• pp.189-193
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• 2010

We investigated the cleavage rate and blastocyst yield for each culture condition to enhance tolerance of cryo-preservation of bovine IVF embryo with relatively lower cryo-tolerance compared to in vivo embryo. The cleavage rate and blastocysts yield for CR1aa, IVMD, IVD, CR1aa+10% FBS were 73.2, 69.3, 72.8, 68.5% and 44.1, 30.8, 33.3, 48.0%, respectively. The values did not differ among each treatments without serum. For embryo vitrification, In vivo and In vitro blastocysts were exposed to VS1(10% glycerin, 0.1 M glucose, 0.1 M sucrose, PEG 1%) for 5 min, and VS2 (10% glycerin, 10% EG, 0.2 M glucose, 0.2 M sucrose, PEG 2%) for 5 min and then VS3 (10% glycerin, 30% EG, 0.3 M glucose, 0.3 M sucrose, PEG 3%) for 1 min. The exposed embryos were then loaded into the 0.25 ml plastic straws and then plunged into liquid nitrogen. The straws were held for period of 1 to 2 weeks before thawing. In embryo viability, no differences in blastocyst re-expansion rates were found between in vivo and in vitro embryos. whereas expansion-BL rates was significantly higher for in vivo-derived embryos (72.7%) when compared to in vitro-derived embryos (51.4%), respectively (P<0.05). In conclusion, our results indicate that combined use of CRIaa culture medium with vitrification might enhance tolerance of cryopreservation for bovine IVF embryo production.

• 한국수정란이식학회지
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• 제25권3호
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• pp.141-144
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• 2010

In vivo embryo produced from Hanwoo donor cows were collected and transferred to Hanwoo recipients. Cows, at random stages of the estrous cycle, received Progesterone Releasing Intravaginal Device (CIDR-plus, InterAg, New Zealand) together with injection of 1 mg estradiol benzoate and 50 mg progesterone, and gonadotropin treatment began 4 day later. For superovulation, a total of 28 mg FSH was intramuscularly injected twice a day in the way of decreasing doses 4 day (5, 5, 4, 4, 3, 3, 2 and 2 mg). Twenty one Hanwoo donor cows were flushed on day 7 of estrus cycle with same FSH and artificial insemination by the same technicians. Embryos were recovered 7 days after the second insemination by flushing the uterus with Embryo Collection Medium. The results obtained were as follows: The rates of transferable embryos were 50.3%, and 78 fresh embryos at morulae and blastocysts stage were transferred into Hanwoo recipients on day 7 of estrus cycle. The pregnancy rates were first embryo transfer 55.6%, 2nd 62.9% and 3rd 57.9%, respectively. In conclusion, These results suggest that CIDR-based superovulation protocol may be effectively used for production of superior Hanwoo embryos. Also, since it seems the condition of recipient cows greatly affect pregnancy rate, it is very important to evaluate recipient for effective cattle production.

• 한국수정란이식학회지
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• 제26권1호
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• pp.79-84
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• 2011

This study was performed to identify the differentially methylated region (DMR) and to examine the mRNA expression of the imprinted H19 gene in day 35 of SCNT pig fetuses. The fetus and placenta at day 35 of gestation fetuses after natural mating (Control) or of cloned pig by somatic cell nuclear transfer (SCNT) were isolated from a uterus. To investigate the mRNA expression and methylation patterns of H19 gene, tissues from fetal liver and placenta including endometrial and extraembryonic tissues were collected. The mRNA expression was evaluated by real-time PCR and methylation pattern was analyzed by bisulfite sequencing method. Bisulfite analyses demonstrated that the differentially methylated region (DMR) was located between -1694 bp to -1338 bp upstream from translation start site of the H19 gene. H19 DMR (-1694 bp to -1338 bp) exhibits a normal mono allelic methylation pattern, and heavily methylated in sperm, but not in oocyte. In contrast to these finding, the analysis of the endometrium and/or extraembryonic tissues from SCNT embryos revealed a complex methylation pattern. The DNA methylation status of DMR Region In porcine H19 gene upstream was hypo methylated in SCNT tissues but hypermethylated in control tissues. Furthermore, the mRNA expression of H19 gene in liver, endometrium, and extraembryonic tissues was significantly higher in SCNT than those of control (p<0.05). These results suggest that the aberrant mRNA expression and the abnormal methylation pattern of imprinted H19 gene might be closely related to the inadequate fetal development of a cloned fetus, contributing to the low efficiency of genomic reprogramming.

• 한국잠사곤충학회지
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• 제51권1호
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• pp.63-67
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• 2013

8개 연구기관의 지역적응 시험을 통해 황보잠은 기존 보급 누에품종인 금황잠보다 화용비율이 높은 강건성 품종으로 대조 품종대비 5령 3일 유충 무게가 3%정도 무겁고, 전견중이 무거워 수량성이 높은 다수성 양친한성황견 품종으로 육성하게 되었다. 이 품종의 일본종계 원종 잠317는 1994년에 Y1, 8013의 교잡으로 2011년에 육성기호 Y9411으로 명명하였다. 암수 모두 무늬가 있는 형잠 품종으로 고치모양은 땅콩형이며 암수고치색깔이 다른 한성황견 품종이다. 중국종계 잠318은 1994년에 Y1, 9134 의 교잡으로 2004년에 육성기호 Y9416으로 명명하였다. 암수 모두 무늬가 없는 희잠 품종으로 고치모양은 타원형이며 암수고치색깔이 다른 한성황견 품종이다. 황보잠은 춘잠기에 적합한 품종으로 사육성적에서 화용 비율이 93.2%, 1만두 수견량이 21.3 kg, 견층중이 54.8 cg으로 대조 품종 금황잠보다 우수하였으며, 또한 실켜기 성적에서 해서율, 생사량이 80%, 4.65kg로 대조 78%, 4.05 kg보다 우수하였다. DNJ(1-Deoxynojirimycin)함량은 황보잠이 대조 금황잠보다 낮았고, 누에 동충하초 생산성에서 감염율은 대조보다 3,9%로 높았고 생체중은 개당 1.32 g으로 대조 1.19 g 보다 무거웠다.

• 한국염색가공학회:학술대회논문집
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• 한국염색가공학회 2011년도 제44차 학술발표회
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• pp.93-93
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• 2011

스포츠웨어 바람막이용도에 사용되는 대부분의 소재는 폴리아미드계 소재이며, 국내에서는 대부분 나일론6 소재를 사용하고 있다. 최근 소비자들의 요구는 점점 경량화되고 있고 야외활동 증가와 더불어 착용감이 우수한 소재를 요구하고 있다. 이러한 시즘에 국내에서 많이 사용되는 나일론6 소재를 이용하여 직물단계에서 경량감과 스트레치성이 발현될수 있는 나일론6 경량 Mechanical 스트레치 직물 개발에 대한 연구를 진행하고자 함이다. 본 연구에서는 직물단계에서 경량감과 스트레치성이 발현될수 있는 나일론6 경량 Mechanical 직물개발을 위해 공중합 나일론6 폴리머와 일반 나일론6 폴리머를 복합방사설비를 이용하여 SIDE BY SIDE POY 26d/6f 원사를 제조한 후, Nip Belt 가연설비로 연신비, 가연 1st 히터온도, 벨트각도 조건에 따라서 제조된 4가지의 나일론6 가연사 DTY 20d/6f와 T사에서 생산되고 있는 나일론6 가연사 DTY 20d/7f를 동일한 직물설계 조건에서 위사방향으로 제직을 실시하였다. 직물설계 조건은 경사를 나일론 DTY 20d/7f, 경사밀도는 2가지, 위사밀도 2가지으로 설계하여 제직하였으며, 이렇게 제조된 직물(생지)를 일반 나일론6 염색가공 공정조건으로 전처리, 염색, 가공, 코팅 공정을 거친후, 신축성 평가를 실시하였다. 동일한 제직조건 및 염색가공 조건에서 제조된 직물의 신축성은 가연조건인 가연 연신비가 높고, 가연 1st 히터온도가 높은 조건에서 높게 나타남을 알 수 있었다. 경사밀도가 낮은 조건이 수축이 많이 진행됨을 알 수 있었으며, 가연조건인 가연 연신비가 높고, 가연 1st 히터온도가 높은 조건에서 수축이 더 많이 이루어짐을 알 수 있었다. 권축률이 높은 가연사 조건이 그만큼 수축이 많이 들어옴을 나타내는 결과로 사료되며, 위사밀도가 낮은 조건이 수축이 좀 더 많이 진행됨을 알 수 있었으며, 위사밀도가 너무 많으면 가연사끼리 상호 수축할 수 있는 정도를 감소시키는 결과로 판단된다. 전처리, 염색가공, 코팅이 완료된 최종 원단의 신축성 평가를 실시한 결과, 비교사로 제직된 최종 원단은 8~10%수준, 가연사 제조조건에 따른 제조된 4가지의 가연사의 신축성은 12~18%의 신축성을 보임을 알수 있었다. 향후, 제직설계 및 염색가공 조건의 다양화로 연구를 진행할 계획이다.

• Reproductive and Developmental Biology
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• 제30권2호
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• pp.87-92
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• 2006

The possibility of producing transgenic embryos expressing the green fluorescence protein (GFP) gene have been evaluated after transfer of exogenous gene into the porcine zygote cytoplasm using the intracytoplasm sperm injection (ICSI) as gene delivery method. For DNA binding to sperm heads, 0.05% Triton X-100 or Lipofectin was used. After injection of the sperm bound to DNA by means of Lipofectin or Triton X-100 triturate, the blastocyst formation rates on day 6 were not significantly different from that of ICSI only group (18.8, 19.2 and 25.3%). In terms of GFP expression, more embryos were in GFP form in Triton X-100 group than in Lipofectin group (40.6 vs 36.4%), while percentage of non-mosaic embryos expressing the GFP gene in all blastomere was higher (P<0.05) in Lipofectin group than in Triton X-100 group (4.2 vs 0.9%). ICSI embryos derived from sperm treated with Lipofectin/DNA complex was transferred into 3 recipients and were collected by uterine flushing on days 5, 7 and 15 after embryo transfer, and then GFP expression was observed by a fluorescence microscopy. Over 26% of the collected embryos were normally expressed GFP gene. These results suggest that foreign gene transfer method with DNA bound sperm caused minimal damage to structure of oocytes that can result to full development of porcine embryos. This was confirmed in this study when the embryos that were transferred after ISCI of DNA bound sperm had a normal development and gene expression until preimplantation.

• Reproductive and Developmental Biology
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• 제32권2호
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• pp.117-121
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• 2008

Our previous study showed that transgenic (TG) pigs harboring human EPO (hEPO) gene have been shown to have reproductive disorders, including low pregnancy rates, irregular estrus cycle and low little size. To investigate these reasons, we assessed estrus behavior (standing response) and plasma $17{\beta}$-estradiol ($E_2$) level, which partly reflect reproductive function, during the estrus cycles after synchronization and superovulation by hormone treatments. Then, we analysed blood composition and expression of hEPO gene in TG pigs. Pigs were injected with PG600. After 10 days, pigs were fed with Regumate porcine for 6 days. Blood samples were collected from jugular vein. Analysis of blood composition and $E_2$ level were measured by Hemavet 950 and $E_2$ ELISA kit, respectively. And, the expression of hEPO gene in reproductive organs was quantitated by real-time RT-PCR. The percentage of estrus behavior in TG was significantly decreased. Hematocrit (HCT), hemoglobin (Hb) concentration and red blood cell (RBC) number were significantly higher in TG than wild type (WT). On the other hand, high expression of hEPO gene in TG was observed in the mammary gland as well as in the uterus. Moreover, plasma $E_2$ level was significantly higher in TG than WT. These results suggest that nonspecific expression of hEPO gene in the other organs of TG may affect blood composition and plasma $E_2$ level, thereby causing reproductive disorders.

• 한방안이비인후피부과학회지
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• 제29권3호
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• pp.106-123
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• 2016

Objectives: This study aimed to verify the classification of atopic dermatitis into the digestive and respiratory disorder on the basis of a literature study.Methods: We searched for the term "atopic dermatitis" in KISS, RISS, NDSL, DBPIA, and OASIS. On further filtering the searched paper further by including the terms "Korean medicine", "literature study" and excluding "acupuncture", we found eight papers relevant to the literature study of atopic dermatitis. The reviewed papers included keywords of Oriental medicine such as Sub-yeol (濕熱), Bi-heo (脾虛), Poong-sub (風濕), Hyeol-heo (血虛). We classified keywords as "digestive system" or "respiratory system" and compared the clinical symptoms of classified Each category keywords.Results: Atopic dermatitis was classified into four categories : acute digestive disorder, chronic digestive disorder, acute respiratory disorder, and chronic respiratory disorder. The four categories of atopic dermatitis showed differences based on region, dermal symptoms, associated symptoms, and affected age group.Conclusions and Discussions: This study suggests a new theory of Atopic dermatitis classification. The theory is similar to that stated in former classification; however, it focuses on the digestive and respiratory disroder. All of atopic dermatitis into digestive disorder show both digestive symptoms and dermal symptoms. However, atopic dermatitis into respiratory disorder shows only respiratory symptoms. Thus, this study can establish a relationship between Western and Oriental medicine's study of atopic dermatitis using keywords such as "digestive disorder" and "respiratory disorder".