• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1995년도 춘계학술대회
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• pp.89-89
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• 1995

Surface epithelial cells isolated from hamster tracheas and grown on a thick collagen gel become a highly enriched population of mucus-secreting cells. Epithelial cells from tracheas of hamsters were collected using enzymatic procedures and cultured under various conditions. The medium used consisted of a 1:1 mixture of medium 199 and Dulbecco's modified Eagle's (DME) medium which was conditioned before use. Insulin, transferrin, hydrocortisone, epidermal growth factor, and extract from bovine hypothalamus were used as supplement. Due to relatively low basal rates of min secretion from in vitro cultures, cultures are generally radiolabeled using $^3$H-glucosamine as a metabolic precursor. The radiolabeled mucinsreleased are quantitated by precipitation with TCA/PTA. Using this cell culture system, we investigated mucin release of goblet cells by altering the media bathing the apical surface of hamster tracheal surface epithelial(HTSE) cells. Acidic media added sulfuric acid caused sigcificant increases in mucin relesse (155${\pm}$20% at pH 4 and 146${\pm}$16% at, pH 5). Ammonium hydroxide also increased mucin release at pH 9.0(156${\pm}$17%) and pH 10(295${\pm}$9%) respectively. This additional mucin release seems to be associated with cell membrane damage as indicated by release of cellular LDH. SP stimulates secretion of mucin in cultured HTSE cells(154${\pm}$16% at 1${\times}$10$\^$-6/M and 165${\pm}$25% at 1${\times}$10$\^$-5/M. PAF at 5${\times}$10$\^$-6/M and 5${\times}$10$\^$-5/M enhanced by HTSE cells in vitro 168${\pm}$34% and 259${\pm}$30% of mucin secretion, respectively. The increase in mucin release by PAF and SP was not secondary to cell damage or necrosis. SP and PAF may be in mediating mucous secretion induced by inflammation irritantion and infection.

• 대한수의학회지
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• 제35권1호
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• pp.187-195
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• 1995

The present study carried out to determine the developmental capacity of bovine oocytes matured in epidermal growth factor(EGF)-containing medium, the developmental competence of bovine embryos using synthetic oviduct fluid(SOF) and the effect of glucose on the development of bovine embryos. In experiment 1, oocytes, obtained from abattoir ovaries, were matured in EGF-containing medium for 24 hours, followed by exposure to Korean native cattle spermatozoa for 18 hours and cultured by utilizing co-culture system with bovine oviduct epithelial cells(BOEC) in TCM199. In experiment 2, early bovine embryos were cultured in SOF with or without BOEC and compared with those in TCM199 with BOEC. In experiment 3, bovine embryos were cultured in the presence or absence of glucose. Seven and ten days after in vitro fertilization, developmental competence of embryos were evaluated. The rate of cleavage was significantly(P<0.05) higher in EGF-containing maturation medium(70.0%) than in control(57.7%). The rates of development to morulae and blastocysts were 30.6% and 23.3% there was no significant difference between them. The rates of in vitro fertilized embryos to morulae and blastocysts cultured in SOF with BOEC(30.4%) and in TCM199 with BOEC(38.0%) were significantly(P<0.01) higher than cultured in SOF without BOEC(13.4%) at seven days after in vitro fertilization. The rates of embryos to blastocysts cultured in SOF with BOEC(29.4%) and in TCM199 with BOEC(35.9%) were significantly(P<0.05) higher than cultured in SOF without BOEC(13.4%) at ten days after in vitro fertilization. The rates of early embryos to morulae and blastocysts cultured in the presence or absence of glucose were 12.2% and 17.5% each other, there was no significant difference between them. The results show that bovine oocytes matured in the presence of EGF can cleave better, SOF with BOEC can replace serum containing complex media, TCM199 with BOEC in bovine embryo culture and glucose have little effect on the culture of early bovine embryos.

• 한국가축번식학회지
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• 제27권2호
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• pp.169-177
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• 2003

본 연구는 착상전 이배체 단위발생 돼지난자를 체외 배양시 우태아혈청 (FBS), 우혈청 알부민 (BSA) 및 상피세포성장인자 (EGF)를 배양액에 첨가하였을 때 배반포, 총 세포수, 세포사멸 및 세포사멸에 관여하는 유전자의 발현을 조사하고자 수행하였다. 0.4% BSA를 배양액에 첨가하였을 때 2세포기 단위발생 난자의 배반포까지의 발달율이 증가되었다(P<0.01). FBS는 배반포의 총세포수를 감소시 켰고 세포사멸을 증가하였다(P<0.01). 그리고 EGF는 BSA가 존재하는 조건하에서 배반포의 총세포수를 증가하였는데 EGF와 BSA가 각각 단독으로 존재할 때는 이런 작용이 없었다. 세포사멸도 이와 이슷한 경향을 보였는데 EGF와 BSA가 각각 존재할 때에는 비처리군과 차이가 없었지만 함께 존재할 때에는 세포사멸을 감소시켰다. RT-PCR의 결과에 의하면 EGF는 BSA가 존재하는 배양액에서 Bcl-xL 유전자의 상대적 발현량을 증가시키고 Bak 유전자의 상대적 발현량에는 영향을 주지 않는 과정을 통하여 세포사멸을 감소시키는것 같다. 반면에 FBS는 Bcl-xL의 발현량을 감소시키고 Bak 유전자의 상대적 발현량을 증가시킨다. 이러한 결과는 세포사멸에 관여하는 유전자의 발현은 배양액의 첨가물에 따라 유의적으로 영향을 받으며, 체외배양시 배아의 초기발달에 관여함을 시사한다.

• 한국응용과학기술학회지
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• 제31권1호
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• pp.120-129
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• 2014

나이를 먹으면 피부의 구조와 생리적 기능이 계속적인 감퇴를 일으켜서 노화된다. 외적요인에 의한 노화는 장기간에 걸친 자외선의 노출로 인한 광노화와 바람, 열, 담배 등이 원인으로 내인성 노화를 촉진시키거나 그 자체도 피부노화를 유발한다. 팔미토일 올리고 펩타이드 또는 세라마이드 올리고 펩타이드는 콜라겐 생산을 자극함으로써 피부의 상층부를 재생하며 아세틸 헥사펩타이드는 피하근육과 피부를 유연하게 하여 주름을 완화하는 성분으로 보톡스 대체 항주름 성분으로 대표적인 고기능성 뷰티성분이다. 대조군과 아세틸 헥사펩타이드 함유 7%, 14%, 20% 성분을 실험 군 A, B, C로 나눠서 주름의 변화, 모공의 변화, 수분량 변화, 과각질의 변화를 분석하였다. 아세틸 헥사펩타이드 함유 성분 분석 결과 통계적으로 주름과 모공, 수분에는 대조군에 비해 영향을 준 것으로 보이며, 과각질의 제거에서는 대조군과 실험군의 값에서 비슷한 결과를 얻을 수 있어 피시험자의 자가 평가의 만족도에서 주름과 모공, 수분에는 대조군에 비해 영향을 준 것으로 보이며, 과각질의 제거에서는 대조군과 실험군의 값에서 비슷한 결과를 얻었다.

• Tuberculosis and Respiratory Diseases
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• 제78권4호
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• pp.315-320
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• 2015

Background: The efficacy of epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) therapy can be measured based on the rate of treatment response, based on the Response Evaluation Criteria in Solid Tumors (RECIST) criteria or progression-free survival (PFS). However, there are some patients harboring sensitive EGFR mutations who responded poorly to EGFR-TKI therapy. In addition, there is variability in the PFS after EGFR-TKI treatment. Methods: We performed a retrospective analysis of the medical records of 85 patients with non-small cell lung cancer, who had achieved a stable disease or better response at the first evaluation of treatment response, after receiving a 2-month course of gefitinib. We calculated the tumor shrinkage rate (TSR) by measuring the longest and perpendicular diameter of the main mass on computed tomography before, and 2 months after, gefitinib therapy. Results: There was a significant positive correlation between the TSR and PFS (R=0.373, p=0.010). In addition, a simple linear regression analysis showed that the TSR might be an indicator for the PFS ($B{\pm}standard$ error, $244.54{\pm}66.79$; p=0.001). On univariate analysis, the sex, histologic type, smoking history and the number of prior chemotherapy regimens, were significant prognostic factors. On multivariate regression analysis, both the TSR (${\beta}$=0.257, p=0.029) and adenocarcinoma (${\beta}$=0.323, p=0.005) were independent prognostic factors for PFS. Conclusion: Our results showed that the TSR might be an early prognostic indicator for PFS in patients receiving EGFRTKI therapy.

• Tuberculosis and Respiratory Diseases
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• 제72권1호
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• pp.15-23
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• 2012

Background: The survival of non-small cell lung cancer (NSCLC) patients with brain metastases is reported to be 3~6 months even with aggressive treatment. Some patients have very short survival after aggressive treatment and reliable prognostic scoring systems for patients with cancer have a strong correlation with outcome, often supporting decision making and treatment recommendations. Methods: A total of one hundred twenty two NSCLC patients with brain metastases who received gamma knife radiosurgery (GKRS) were analyzed. Survival analysis was calculated in all patients for thirteen available prognostic factors and four prognostic scoring systems: score index for radiosurgery (SIR), recursive partitioning analysis (RPA), graded prognostic assessment (GPA), and basic score for brain metastases (BSBM). Results: Age, Karnofsky performance status, largest brain lesion volume, systemic chemotherapy, primary tumor control, and medication of epidermal growth factor receptor tyrosine kinase inhibitor were statistically independent prognostic factors for survival. A multivariate model of SIR and RPA identified significant differences between each group of scores. We found that three-tiered indices such as SIR and RPA are more useful than four-tiered scoring systems (GPA and BSBM). Conclusion: There is little value of RPA class III (most unfavorable group) for the same results of 6-month and 1-year survival rate. Thus, SIR is the most useful index to sort out patients with poorer prognosis. Further prospective trials should be performed to develop a new molecular- and gene-based prognostic index model.

• 약학회지
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• 제41권6호
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• pp.737-748
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• 1997

Distribution of rhEGF in the skin, plasma and several organ tissues following topical application of $^{125}I-rhEGF$ (0.4${\mu}$Ci) solution in 25% Pluronic F-127 on 154$mm^2$ normal and damaged (burned and stripped) skins of hairless mice was examined. The radioactivity in the stripped skin tissues increased as a function of time, and was 10-20 times higher than that in the normal and burned skins. The fractions of intact drug in the skin tissues were 40-60% for the normal and burned skins, and 60-80% for the stripped skin. It indicates that the stratum corneum layer behaves as a barrier for the dermal penetration of the drug. The radioactivity in the plasma was much higher for the stripped skin than for the normal and burned skins. However, the concentration of intact drug in the stripped skin was comparable to those in the normal and burned skins indicating most severe degradation (or metabolism) of the drug in the stripped skin. As a result, the fraction of intact drug in the plasma was lowest for the stripped skin (<10%). Body organ distribution of the drug was much higher for the stripped skin. The concentration in the stomach. Both in total radioactivity and intact drug, showed more than 10-times higher value than in the other organs (liver, kidney and spleen). The fraction of intact drug in each organ tissue was below 10-20%. And generally lowest for the stripped skin. The lowest fraction of the drug for the stripped skin could not be explained by the activity of the aminopeptidases in the skin since it was lower for the stripped skin than for the normal skin. Thereover, the fraction of intact drug appears to be determined by the balance between dermal uptake and systemic elimination of the drug, for example. The mechanism of dermal uptake of rhEGF was examined by topical applying 200${\mu}$l of 25% Pluronic F-127 solution containing 0.4 ${\mu}$Ci of $^{125}I-rhEGF$ and 0.14${\mu}$Ci of $^{14}C$-inulin (a marker of passive diffusion). The radioactivity of $^{125}I-rhEGF$ at each sampling time point (0.5, 1, 2, 4 and 8hr) was correlated (p<0.05) with the corresponding radioactivity of $^{14}C$-inulin. It appears to indicate the rhEGF may be uptaken into the skins mainly by the passive diffusion. This hypothesis was supported by the constant specific binding of EGF to the skin homogenates regardless of the skin models. Receptor mediated endocytosis (RME) appears to contribute negligibly, if any, to the overall uptake process.

• Applied Microscopy
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• 제29권2호
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• pp.163-175
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• 1999

본 실험은 화상을 입은 생쥐에서 SQ의 치유 효과를 알아보기 위함에 있다. 실험군은 7개군으로 구분하였다; 대조군, 화상군, 그리고 화상을 입헌 후 SQ를 처치한 군. 대조군을 제외한 모든 군은 등 부위에 2도 화상을 입혔다. SQ를 처치한 모든 군은 하루에 한 번씩 10초 간격으로 순수한 SQ 3방울을 도포하였다. 화상 후 10일 동안 조직학적 및 미세구조적 변화를 광학 현미경과 전자현미경으로 관찰하였다. 광학현미경으로 관찰한 결과, 화상군의 표피층의 모든 부분은 SQ를 처치한 군보다 더 심한 상처를 입었다. SQ를 처치한 6째와 10일째 군에서, 기저층은 분화가 활발하였고, 가시층의 각화세포는 수적으로 매우 증가하였다. 전자현미경으로 관찰한 결과, SQ를 처치한 모든 군의 기저층에서 세포분열이 화상군에서보다 더 활발하게 나타났다. 특히, SQ를 처치한 10일군에서는 이상적인 결과를 얻을 수 있었다; 대조군보다 더 두꺼운 가시층과 많은 각화세포, 뚜렷한 세포간교, 그리고 멜라닌세포를 포함하는 튼튼한 기저층. 이와 같은 결과로 보아 SQ은 표피성장인자(EGF)를 활성화시키고, 유해산소의 제거 역할을 하며, 막계에 에너지원을 제공할 가능성이 있다. 이 실험의 결과는 SQ가 화상 치료에 특수한 효과가 있다고 사료된다.

• 한국수정란이식학회지
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• 제11권2호
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• pp.125-135
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• 1996

The present study was carried out to investigate the effects of cryoprotectants, equilibration step, freezing rate, culture condition following in vitro fertilization, and age and development stage of embryo by freezing with conventional slow freezing and vitrification on survival of frozen-thawed Korean native cattle(KNC) blastocysts produced in vitro. The KNC blastocysts produced in vitro were equilibrated in 1.8M ethylene glycol or 1.4M glycerol and cooled from -6$^{\circ}C$ to -35$^{\circ}C$ at -0.3$^{\circ}C$ or -O.6$^{\circ}C$ /minute. When equilibrated in 1.8M ethylene glycol, survival rate of fiozen4hawed blastocysts was sarne in both -0. 3$^{\circ}C$ /min and -0.6$^{\circ}C$ /min cooling rate(71.4%). With the equilibration in 1.4M glycerol, survival rate was higher in -0.3$^{\circ}C$ /min(63.6%) than in -0.6$^{\circ}C$ /min cooling rate(53.8%). For vitrification of the KNC blastocysts produced in vitro, they were equilibrated in 2-step or 3-step exposure to vitrification solution(25% ethylene glycol + 25% glycerol). Survival rate was sirilar in both 2-step(45.0%) and 3-step exposure(47.4%). According to culture condition following in vitro fertilization, higher survival rate was obtained for blastocysts co-cultured with bovine oviductal epithelial cell(BOEC, 77.3%) than for those cultured with epidermal growth factor(EGF, 65.7%) or for those co-cultured with BOEG + EGF (54.8%). According to embryo age and development stage, higher survival rate was obtained for 7-day ernbryos(70.0%) than 8-day(56.8%) or 9-day(20.0%) for blastocyst stage and obtained for 8-day embryos(74.3%) than 7-day(62.5%) or 9-day(42.9%) for exponded blastocyst. In surnmary, higher survival rate of frozen4hawed KNC blastocysts produced in vitro were obtained by using ethylene glycol for cryoprotectant and -0.3$^{\circ}C$ /min for cooling rate. And higher survival rate were obtained with co-culture with BOEC for culture condition following in vitro fertilization and with 7-day blastocyst or 8-day expanded blasto cyst for embryo age and development stage.

• 한국수정란이식학회지
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• 제22권4호
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• pp.245-249
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• 2007

The aim of present experiment was to examine hatching rate as in vitro indicator of viability of porcine embryos before early stage embryo transfer such as zygotes or 2-cell stage embryos. Cumulus-oocyte complexes (COCs) collected from ovaries were matured in North Carolina State University 23 (NCSU-23) containing 10% porcine follicular fluid (pFF), 10 ng/ml epidermal growth factor (EGF), $10{\mu}g/ml$ follicle stimulating hormone (FSH), $35{\mu}g/ml$ luteinizing hormone (LH), and 1mg/ml cysteine. After 24 hours, the COCs were transferred to the same medium without hormones. After 65h of maturation, oocytes were exposed to phosphate buffered saline (PBS) with 7% ethanol (v/v) for 7 minutes, and then the oocytes were washed and cultured in tissue culture medium (TCM) 199 containing 5 ug/ml cytochalasin B for 5h at $38.5^{\circ}C$ in an atmosphere of 5% $CO_2$ and 95% air with high humidity. After cytochalasin B treatment, the presumptive parthenotes were cultured in porcine zygote medium (PZM)-5 and cleavage of the parthenotes was assessed at 72h of activation, Normally cleaved parthenotes were cultured for an additional 8 days to evaluate their ability to develop to blastocyst and hatching stages. The fetal bovine serum (FBS) were added at Day 4 or 5 with concentrations of 2.5, 5 or 10%. The blastocyst rates were ranged within $39.1{\sim}70%$ in each treatment. However hatching rate was dramatically decreased in non-addition group. In this experiment, embryo viability in female reproductive tract may be estimated before embryo transfer with in vitro culture adding FBS by hatching ability.