• Archives of Plastic Surgery
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• 제39권6호
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• pp.593-599
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• 2012

Background This study aimed to investigate the possibility of isolating mesenchymal stem cells (MSCs) from human thigh adipose tissue and the ability of human thigh adipose stem cells (HTASCs) to differentiate into hepatocytes. Methods The adipose-derived stem cells (ADSCs) were isolated from thigh adipose tissue. Growth factors, cytokines, and hormones were added to the collagen coated dishes to induce the undifferentiated HTASCs to differentiate into hepatocyte-like cells. To confirm the experimental results, the expression of hepatocyte-specific markers on undifferentiated and differentiated HTASCs was analyzed using reverse transcription polymerase chain reaction and immunocytochemical staining. Differentiation efficiency was evaluated using functional tests such as periodic acid schiff (PAS) staining and detection of the albumin secretion level using enzyme-linked immunosorbent assay (ELISA). Results The majority of the undifferentiated HTASCs were changed into a more polygonal shape showing tight interactions between the cells. The differentiated HTASCs up-regulated mRNA of hepatocyte markers. Immunocytochemical analysis showed that they were intensely stained with anti-albumin antibody compared with undifferentiated HTASCs. PAS staining showed that HTASCs submitted to the hepatocyte differentiation protocol were able to more specifically store glycogen than undifferentiated HTASCs, displaying a purple color in the cytoplasm of the differentiated HTASCs. ELISA analyses showed that differentiated HTASCs could secrete albumin, which is one of the hepatocyte markers. Conclusions MSCs were islolated from human thigh adipose tissue differentiate to heapatocytes. The source of ADSCs is not only abundant abdominal adipose tissue, but also thigh adipose tissue for cell therapy in liver regeneration and tissue regeneration.

• 대한임상검사과학회지
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• 제38권2호
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• pp.111-116
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• 2006

One of the cardinal findings of the renal diseases is proteinuria, which appears in the early phase of kidney diseases and is very important in diagnosis, prognosis and decision making in the treatment process and results of the treatment. The study subjects were 126 patients who visited the nephrology department of Kangbuk Samsung Hospital. Serum was requested for urine protein electrophoresis. Total protein was measured with Bayer Advia 1650 (Biuret). Quantitation of each fraction was done by multiplying the percentage of each fraction by the total protein. Serum creatinine and BUN were also measured with Bayer Advia 1650 (Jaffe and Urease). Serum protein EP was done with REP(rapid electrophoresis) using Helena Kit reagents (REP Ultra SPE Kit, Ponceau S stain, Acetic acid, Methanol, EP Control). Concentrated urine was used for urine protein EP. The SPSS package was used for statistics analysis. Percentage and quantitation of the level of albumin in renal diseases were significantly lower than those in healthy controls. Total protein was correlated with albumin. In terms of proportion, ${\alpha}1$-globulin, ${\alpha}2$-globulin, ${\beta}$-globulin, and ${\gamma}$-globulin fractions were increased in the disease group. But, in the quantified level, ${\alpha}2$-globulin was increased and ${\beta}$-globulin and ${\gamma}$-globulin were decreased. ESRD patients showed an increased secretion of high molecular proteins in urine protein EP. A decreased level in serum total protein correlated with the decreased level of serum albumin and the total amount of urine total protein. This study revealed the variety in the level of serum and urine proteins and their subgroups by EP.

• Applied Biological Chemistry
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• 제42권3호
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• pp.229-234
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• 1999

사람 간세포 유래의 배양 세포인 HepG2 세포의 지질 합성과 분비에 미치는 linoleic acid(LA, 18 : 2 n-6) 및 우혈청알부민 (bovine serum albumin: BSA) 첨가 농도의 영향에 대하여 검토하였다. 간세포는 DMEM배지(기본 배지)에 0.2 mM LA을 첨가한 배지 및 LA와 BSA(0.2-1.0%)를 첨가한 배지(LA+BSA 배지)에서 배양하였다. 각 지질의 동위원소 표적에는 $[^{14}C]acetate$를 이용하여 6시간 배양후의 지질합성과 분비를 측정하였다. 그 결과, 기본 배지중에 LA의 첨가는 콜레스테롤의 $[^{14}C]acetate$ 표적량을 저하 시켰다. 한편, LA배지에 BSA의 첨가에 의해, 총 콜레스테롤의 $[^{14}C]acetate$ 표적량은 증가하는 경향을 나타내었다. LA+BSA 배지에서 간세포의 총 콜레스테롤 변동은 유리형 콜레스테롤에의 표적량 증가에 기인하는 것으로 나타났다. LA배지에 BSA를 첨가 하였을때 콜레스테롤 분비가 증가 하였는데, 이는 지질의 분비과정에 BSA가 관여하는 것을 시사하는 것이다. 간세포내 총 지질에의 $[^{14}C]acetate$ 표적량은 각 군간의 유의치는 인정되지 않았다. 그러나, LA 배지에의 BSA 첨가는 $[^{14}C]acetate$표식 지방산의 중성지질 획분에의 표적량은 증가하고, 인지질 획분에 표적량은 감소하여, 양지질의 합성에는 상반되는 결과를 나타내었다. $[^{14}C]acetate$ 표적 중성지질, 인지질 및 유리지방산의 분비는 LA배지에 비교해 LA+BSA 배지에서 현저하게 증가하였다. 이상의 결과에서, 사람 간배양 세포에서 LA와 BSA는 각각 지질대사에 다른 영향을 미치고, BSA 농도는 리포단백질 분비에 영향을 미치는 것으로 시사된다.

• 동국한의학연구소논문집
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• 제8권1호
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• pp.145-158
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• 1999

본 실험은 사군자탕합창출지유탕(四君子湯合蒼朮地楡湯)의 만성설사(慢性泄瀉)에 미치는 영향을 조사하기 위해 시행되었다. 실험을 위해 흰쥐에 3주일간 castor oil과 대황(大黃)분말을 병행 투여하여 1주일후부터 지속적인 설사를 보인쥐를 선정, 만성설사의 모델로 삼았다. 이를 대조군과 실험군으로 구분하여 위장관 기능과 영양상태를 비교하였고, 개선여부를 알아보기 위해 아무런 처치를 받지 않은 정상군을 검체 체취에 사용하였다. 실험군과 대조군에서 검액 투여전 모두 위장관 기능과 영양상태가 감소한 상태였으며, 검액을 투여한 후 실험군에서 대조군에 비해 위장관 기능과 영양상태가 개선된 경향을 보였는데, 특히 gastrin과 total lipid, 그리고 hematocrit 수치가 유의성 있게 나타나 사군자탕합창출지유탕(四君子湯合蒼朮地楡湯)이 만성설사(慢性泄瀉)에 효과가 있는 것으로 사료된다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제3권2호
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• pp.96-102
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• 1998

Primary hepatocytes of small animals such as rat and rabbit were often used for the study of extracorporeal liver support systems. Freshly isolated rat hepatocytes form spheroids within tow days when cultivated as suspension in spinner vessels. These spheroids showed enhanced liver specific functions and more differentiated morphology compared to hepatocytes cultured as monolayers However, shear stress caused by continuous agitation deteriorated spheroids gradually. In this work we immobilized spheroids to prolong liver specific activities. First, hepatocyte spheroids were suspended in collagen solution containing calcium chloride and then dropped into alginate solution. A thin layer of calcium alginate was formed around the droplet and then was removed after the inner collagen was gelled by treatment of sodium citrate buffer. Spheroids embedded in collagen-gel bead maintained liver specific functions such as albumin secretion rate longer than hepatocyte spheroids exposed to shear stress. Therefore, we suggest that this immobilization technique may offer an effective long-term hepatocyte cultivation and facilitase the development of a bioartificial liver support device.

• Journal of Acupuncture Research
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• 제22권4호
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• pp.189-195
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• 2005

Objectives : This study was to evaluate the effectiveness of Bee Venom acupuncture therapy on sprain of C-spine by randomized controlled double blinding method. Methods : A randomized double blinding study of bee venom acupuncture therapy was conducted on sprain of C-spine. We divided sprain of C-spine patient into 2 groups: one group with both acupuncture and saline acupuncture therapy, and another group with both acupuncture and bee venom acupuncture therapy. To estimate the efficacy of treatment that applied for two groups, we used visual analog scale(VAS) and neck disability index(NDI). We compared the VAS and NDI score of two groups statistically. Results : STZ induced increase of serum creatinine, BUN and albumin secretion were lowered by AD-1 treatment.

• 대한약리학회지
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• 제10권2호
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• pp.39-42
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• 1974

Despite a considerable amount of investigation there continues to be disagreement concerning the proteins present in human seminal plasma. Recently their identification has assumed a greater importance following evidence that infertility in men and women may have an immunological cause (Katsh, 1959; Quinlivan, 1969). Seminal plasma is composed of fluids secreted by the prostate, seminal vesicles, ampullae, ducti deferentes, bulbourethral (Cowper's) glands, urethral(Littre's) glands and the epididymes. Prostatic juice, one of the major components of seminal plasma, has an important role in secretion of acid phosphatase and prostaglandin. A few studies have been reported of human prostatic juice, since, in human subjects, there were some problems in studying prostatic juice due to quite small amount of secretion and possibility of contamination with fluids from the seminal vesicles and ejaculatory ducts. The purpose of the present study was to determine the basic components of proteins in human prostatic juice. Prostatic juice was obtained from normal healthy man of $20{\sim}30\;year-old$ by massage of the prostate, and protein components were separated by means of disc electrophoresis. The results are summarized as follows; 1) Total numbers of protein fractions of normal human serum and prostatic juice are $14{\sim}18$ bands and $9{\sim}12$ bands, respectively. Prostatic juice produces two deeply staining bands which appear similar to those formed by $beta-_1$ globulin and albumin. 2) $Alpha-_1$ globulin area in the fractions of prostatic juice shows 4 bands and one more band is found than that of serum. On the other hand, the fractions of immunoglobulin and $alpha-_2$ globulin areas are eight in serum and it has three bands more than that of prostatic juice. 3) $Alpha-_1$ globulin area in the prostatic juice is more deeply stained than that of serum. In contrast with $alpha-_1$ globulin area, immunoglobulin and $alpha-_2$ globulin areas in the prostatic juice show weaker staining than serum.

• 한국가금학회지
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• 제33권1호
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• pp.49-55
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• 2006

이 연구는 시호의 첨가가 육용종계의 생산성, 혈액의 성상, 호르몬 분비에 미치는 영향을 구명하기 위해서 8주간 실시하였다. 총 800수의 20주령 육용종계를 4개 처리구에 각각 200수씩 배치하였고, 시호는 사료에 0, 0.5, 0.1, 0.2% 수준으로 첨가하여 급여하였다. 기초사료의 영양소 함량은 육성기와 산란기로 구분하여 각각 CP 15.0%, ME 2,900 kcal/kg 수준과 CP 17.5%, ME 2,800 kcal/kg 수준으로 하였다. 시호를 육성후기에($20{\sim}24$주) 급여하여 육용종계의 증체량을 조사한 결과, 처리구간에 차이는 없었으나 산란초기($24{\sim}28$주)의 산란율과 산란량은 시호 급여구에서 대조구에 비하여 유의적으로 증가하였다(p<0.05). 시호의 첨가에 따른 육용종계의 혈중 변화를 관찰한 결과, 혈중 콜레스테롤 농도의 경우 대조구에 비하여 시호 2.0% 급여구에서 가장 낮은 수치를 나타내었다(p<0.05). 이와는 다르게 혈중 중성지방 농도에서는 시호의 급여 효과는 관찰되지 않았다. 반면, 혈중 헤마토크릿, 포도당, 칼슘, 인 및 마그네슘 농도에서 시호의 첨가 효과는 관찰할 수 없었다. 혈중 총 단백질과 알부민 함량 또한 시호 급여에 따른 변동은 관찰할 수 없었으나 알부민/글로불린 비율은 시호 급여구가 유의하게 감소하였다(p< 0.05). 혈중 GGT, AST, ALT 및 BUN 농도에서도 시호 급여구와 대조구간의 차이는 관찰되지 않았다. 28주령 육용종계의 혈중 에스트로겐 농도는 시호 급여구가 전체적으로 증가하는 양상을 보였으며, 그 중 시호 0.5% 급여구의 에스트로겐 농도가 현저하게 증가하였다(p<0.05). 또한 IGF-I 농도는 시호 급여구가 대조구와 비교하여 유의하게 증가하였다(p<0.05). 이 연구의 결과 육용종계 사료에 시호의 첨가는 생산능력을 개선하고, 혈중 콜레스테롤 농도와 A/G 비율을 감소시켰으며, 에스트로겐과 IGF-I 분비를 증가시켰다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권2호
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• pp.99-105
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• 2000

Cell-Cell interaction and the extracellular matrix (ECM) are belisved to play essential roles during in vitro culturing of primary hepatocytes in the control of differentiation and in the maintenance of tissue spcific functions. The objective of this study was to examine the effects of degree of cell-cell contact (DCC) on liver sperific function of rat promary hepatocytes. Hepatocyte aggregates with various with various degrees of cell-cell contantact, I. e., dispersed cell, longish aggregate, rugged aggregate, and smooth spheroid were obtained at 1, 5-6, 15-20, and 36-48 hrs, respectively in suspension cultures grown in spinner flasks embedded in Caalginate bead and collagen gel in order. The may result from mass transfer limitation and shear damage caused by agitation during aggregation. The rugged aggregate showed a higer viability and albumin secretion rate than the dispersed cells or the other aggregates. This result indicates the possible enhancement of a bioartificial liver's (BAL) performance using primary hepatocytes and the reduction in time to prepare a BAL through optimization of the immobilization time.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 춘계학술발표대회
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• pp.141-144
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• 2000

Highly open porous polymer matrices are required for high density cell seeding, efficient nutrient, and oxygen supply to the cells cultured in the three dimensional matrices. However, there are severe problems of mass transfer limitations within the cell/scaffolds culture system. Thus we hypothesize that continuos-flow culture conditioning of cells with the scaffolds may improve the cell viability and the differentiated function. In this study, we fabricated porous PLGA scaffolds by using gas-foaming/salt-leaching method as previous described. Viscous PLGA gel paste contains ammonium bicarbonate particulates, acting as a gas-foaming agent as well as a salt-leaching porogen, were cast into Teflon mold and dried. Ammonium bicarbonate salt upon contact to an acidic aqueous solution evloves gaseous ammonia and carbon dioxide by itself. And we conjugated galactose moiety [AGA; $N-(aminobuty1)-O-{\beta}-D-galactopyranosyl-(1{\rightarrow}4)-D-glucoamide]$ to the terminal end group of a PLGA to increase the cell adhesion and matain the differentiated function of hepatocytes. Cell-seeded scaffolds were secured in a flow bioreactor chamber and exposed to continuous flow at 5 ml/min. As a result of our study, the high yield of hepatocytes attachment was accomplished by increasing the concentration of PLGA-AGA conjugate in polymer scaffolds and cells in the scaffolds under continuos flow condition maintained a high level of viability and albumin secretion rate of cultured hepatocytes showed a higher level that of control groups.