Reconstruction of Rabbit Corneal Epithelium on Lyophilized Amniotic Membrane Using the Tilting Dynamic Culture Method

Ahn, Jae-Il,Lee, Doo-Hoon,Ryu, Yang-Hwan,Jang, In-Keun,Yoon, Mun-Young,Shin, Youn Ho,Seo, Young-Kwon,Yoon, Hee-Hoon,Kim, Jae-Chan,Song, Kye-Yong,Yang, Eun-Kyung,Kim, Ki-Ho,Park, Jung-Keug Blackwell Publishing Inc 2007 Artificial Organs Vol.31 No.9

<P>Abstract: </P><P>Rabbit corneal epithelium was reconstructed using tilting dynamic culture with a self-manufactured, amniotic membrane (AM) supporter and a lyophilized amniotic membrane (LAM). Rabbit corneal epithelial (RCE) cells were cultured and cryopreserved after isolation from the limbus. The second- and third-passage RCE cells were plated onto the epithelial side of the LAM of Ahn's AM supporter. Two days later, the air–liquid interface culture was maintained with third-passage RCE cells for 6 days and second-passage corneal epithelial cells for 9 days. The average viability of thawed RCE cells, assessed using trypan blue dye exclusion, was 77.42%. The reconstructed corneal epithelium was characterized by histological (hematoxylin and eosin) and immunohistochemical staining (proliferating cell nuclear antigen) for light microscopy, and by reverse transcriptase-polymerase chain reaction, glucose assay, and transmission electron microscopy. The basal layer of the reconstructed corneal epithelium was well formed, and the epithelium was tightly constructed due to the increase in cell proliferation and differentiation caused by the tilting dynamic culture, as opposed to static culture. Tilting dynamic culture was useful for the reconstruction of the epithelium using easily damaged epithelial cells and resulted in more stratum cell layers. Moreover, cytokeratin (CK3) mRNA expression in tilting dynamic cultured third-passage RCE cells seeded onto AM was greater than in static cultured third-passage RCE cells. The morphology of the reconstructed corneal epithelium on LAM by tilting dynamic culture for 9 days resembled that of the skin epidermis. This was thought to be because the tilting dynamic culture not only accelerated the proliferation and differentiation of cells by physical or mechanical stimulation, but also ensured that the supply of medium was delivered to the basal cells more efficiently. Thus, the reconstruction of the corneal epithelium using LAM and tilting dynamic culture was considered to be a good in vitro model for autologous or allogeneic transplantation of corneal epithelium and skin epidermis in patients with damaged epithelia. </P>

비만을 동반한 위암 환자에서 두 병을 같이 수술 치료한 경험(증례)

최경현,윤기영,문형환,신연명,서경원,안수미,송윤미,석정희,정경연,이은하 고신대학교의과대학 2008 고신대학교 의과대학 학술지 Vol.23 No.3

Obesity is growing problem in Korea. We had a case of bariatric surgery during gastric cancer operation. Patient was 29 year old Korean lady with early gastric cancer located in the lesser curvature side of the middle 1/3 of the stomach. Her body weight was 89 kg, height 163 cm, and thus body mass index was 33.5 kg/m2. Preoperative blood pressure was 130/90 mmHg, hemoglobin 12.9 g/dL, total lymphocyte count 3,280/mL, serum albumin 4.3 g/dL, CEA 1.1 ng/mL, CA 19-9 9.1 ng/mL, and alpha fetoprotein 2 ng/mL. Another associated disease was right thyroid follicular neoplasm. The chest X-ray was normal. After IM injection of 2,500 units of heparin 30 minutes before the induction of general anesthesia, she received curative subtotal gastrectomy and Roux en Y gastrojejunostomy when a malabsorption loop of 120 cm jejunum was incorporated between the Treitz ligament and the end to side jejuno-jejunostmy site at May 29th 2006. Her postoperative course was uneventful except a minor wound seroma and the elevations of serum amylase(up to 4 folds) and lipase(up to 2 folds). She lost her body weight 9 kg in 5th, 21 kg in 7th, and 27 kg in 10th postoperative months to became 61.5 kg. On follow up exams in July 2008, she gained 4 kg to overcome her weakness and fatigue. From the above result, the incorporation of a malabsorption loop during reconstruction phase of gastric cancer operation was a good option for obese gastric cancer patients especially in cases of early gastric cancer.

강박증 환자에서 신경 심리 검사를 통한 전두엽 기능에 관한 연구

안경흡,김명선,김연수,박은희,박선희,신민섭,류인균,권준수 大韓神經精神醫學會 2000 신경정신의학 Vol.39 No.3

연구목적 : 강박증 환자에서 비언어적 기억의 장애, 실행능력(executive function)의 장애, 시공간능력의 장애등의 신경 인지 기능의 장애 및 뇌 영상 연구에서 전두엽-기저핵 신경로의 기능장애가 있다는 결과들이 보고되어 왔다. 저자들은 전두엽의 기능을 반영하는 것으로 알려진 신경 심리 검사를 통하여 강박증 환자의 전두엽의 기능적 변화의 유무 및 기타 인지기능에 미치는 영향에 대해서 평가하고자 하였다. 방 법 : 서울대학교병원 강박증 클리닉에 내원한 32명의 강박증 환자군과 28명의 정상 대조군을 대상으로 강박증상의 중증도에 대한 평가와 함께 한국판 단축형 지능검사(Short-form of K-WAIS)와 주로 전두엽 기능을 평가하는데 민감한 것으로 알려진 신경 인지 기능 검사군(위스콘신 카드 분류 검사, Rey-Os-terrieth Complex Figure test, Controlled Oral Word Association test, 선로 잇기 검사, Luria-Nebraska 신경심리 바테리 중 언어학습능력검사)을 시행하였다. 이 검사들의 결과를 연령과 교육연수를 통제한 후에 ANCOVA로 분석하였다. 결 과 ; 강방증 환자군은 범주적 언어 유창성 점수(ANCOVA, F=15.07, df=58, p<0.001), Rey-Os-terrieth Complex Figure test의 즉각 회상 점수(ANCOVA, F=6.33, df=57, p<0.015), Rey-Os-terrieth Complex Figure test의 지연 화상 점수(ANCOVA, F=5.33, df=57, p<0.022)가 정상 대조군에 비하여 유의하게 저하되어 있었다. 결 론 : 이 결과는 강박증 환자들이 전두엽의 특징적인 기능으로 알려진 실행기능보다는 비언어적 기억능력의 장애 및 언어능력의 장애를 가지고 있음을 시사한다. Objective : Non-verbal memory deficits, impairments in executive function and deficits in visuospartial functions have been repeatedly reported in subjects with obsessive-compulsive disorder(OCD). The present study aimed to evaluate the frontal lobe function using neuropsychological test in subjects with OCD and normal control. Methods : A battery of neuropsychological test(Wisconsin Card Sorting test, Rey-Osterrieth Complex Figure test, Controlled Oral Word Association test, Trail Making test, Verbal Learning test) reflecting frontal lobe function was administrated to 32 OCD patients and 28 healthy comparison subjects. Result : There were no significant differences in age, years of education, or estimated IQ between the groups. Scores in category fluency, immediate recall and delayed recall of Rey-Osterrieth Complex Figure test in OCD subjects were significantly lower than those of healthy comparison subjects(ANCOVA, F=15.07, df=58, p<0.001 ;ANCOVA, F=6.33, df=57, p<0.015 ; ANCOVA, F=5.33, df=57, p<0.022, respectively). Conclusion : OCD patients had selective deficits in task involving non-verbal memory and categorical word fluency relative to healthy comparisons.

입원 환자 영양검색 전산 프로그램 개발

안수미,윤기영,신연명,최경현,임근승,장문경,송윤미,공은희 고신대학교의과대학 2008 고신대학교 의과대학 학술지 Vol.23 No.2

Background : About 30% of patients in the hospital are under nourishment. A large portion of people are undernourished when they are admitted to the hospital and in the majority of these, undernutrition develops further while in hospital. Patient-Generated subjective global assessment(PG-SGA), a method of nutritional assessment based on clinical judgment, has been widely used to assess the nutritional status of adults for both clinical and research purposes. Modified PG-SGA can be used as a nutrition screening tool, and a comprehensive nutrition assessment is desirable for those who are malnourished. The purpose of this study is to develop a simple, reliable and valid nutrition screening program(NSP). Methods : We applied the nutrition screening program(NSP) to adult inpatients who are at risk of malnutrition using the Modified Patient-Generated subjective global assessment(PG-SGA) and objective assessment(electrical medical record data). Results : Therefore simple and beneficial NSP is developed for patients with severe malnutrition. Using NSP. nutritional information of the severe malnutrition patient should be shared with the physicians and they should be taken care of by clinical dietitians to improve their nutritional status. Conclusion : The Modified PG-SGA and nutrition screening tool are simple, reliable and valid nutrition screening program(NSP).

결정형 규산분진에 폭로된 섬유모세포의 자가증식 : Evaluation by H₂O₂and PDGF-AA 와 TGFβ

안병용,김경아,문제혁,정진숙,김은경,임영 대한산업의학회 2000 대한직업환경의학회지 Vol.12 No.2

Objectives : The aim of this study is to find out the activity of autoproliferation of rat fibroblast exposed to crystalline silica and the role of mediators secreted from rat fibroblast. Methods : The effect of α-quartz on production of growth factor (platelet-derived growth factor-AA and transforming growth factorβ) from rat fibroblasts were evaluated by ELISA and immunocytochemical analysis. Gene expression of these growth factors in rat fibrobast exposed to crystalline silica was evaluated by RT-PCR. Furthermore, fibroblast proliferation by culture supernatant of rat fibroblast was assayed by the neutral red test. Results : The amounts of H2O2 and growth factors synthesized in rat fibroblasts were significantly increased by the stimulation of crystalline silica (α-quartz) , which showed the dose-dependent manner to the concentration of α-quartz with the maximum response at the dosage of 100 ㎍/㎠. The result of RT-PCR demonstrated that α-quartz induced gene expression of PDGF-AA and TGFβ in rat fibroblast. We also found that supernatant of α-quartz-cocultured rat fibroblast induced a significant proliferation of fibroblast. Conclusion : Crystalline silica directly induce functional change In fibroblast such as increased release of reactive oxygen species and growth factors. The products of these functional change promote fibroblast proliferation via autocrine loop.

항암제 AG60이 흰생쥐 위점막 상피세포의 DNA합성에 미치는 영향 : 자기방사법적연구

박경호,안의태,고정식,하상선,한영복,정영신,홍은경,유보림,김상건,이경영,강종구 대한해부학회 1999 Anatomy & Cell Biology Vol.32 No.1

A Comparison of Lyophilized Amniotic Membrane with Cryopreserved Amniotic Membrane for the Reconstruction of Rabbit Corneal Epithelium

Ahn Jae-Il,Jang In-Keun,Lee Doo-Hoon,Seo Young-Kwon,Yoon Hee-Hoon,Shin Youn-Ho,Kim Jae-Chan,Song Kye-Yong,Lee Hee-Gu,Yang Eun-Kyung,Kim Ki-Ho,Park Jung-Keung The Korean Society for Biotechnology and Bioengine 2005 Biotechnology and Bioprocess Engineering Vol.10 No.3

Many researchers have employed cryopreserved amniotic membrane (CAM) in the treatment of a severely damaged cornea, using corneal epithelial cells cultured on an amniotic membrane (AM). In this study, two Teflon rings were made for culturing the cells on the LAM and CAM, and were then used to support the AM, which is referred to in this paper as an Ahn's AM supporter. The primary corneal epithelial cells were obtained from the limbus, using an ex-plantation method. The corneal epithelium could be reconstructed by culturing the third­passage corneal epithelial cells on the AM. A lyophilized amniotic membrane (LAM) has a higher rate of graft take, a longer shelf life, is easier to store, and safer, due to gamma irradiation, than a (AM. The corneal epithelium reconstructed on the LAM and (AM, supported by the two­Teflon rings, was similar to normal corneal epithelium. However, the advantages of the LAM over that of the (AM make the former more useful. The reconstruction model of the corneal epithelium, using AM, is considered as a good in vitro model for transplantation of cornel epithelium into patients with a severely damaged cornea.

사상체질과 신체형태지수와의 연관성

박경아,김희진,허경석,안수경,이성호,이종은,이원택 대한췌담도연구회 2000 대한췌담도연구회지 Vol.5 No.4

심폐소생술의 순환회복 예측인자로서 호기말 이산화탄소 분압의 유용성 및 의의 : 각 심정지 원인에 따른 분석

어은경,안기옥,김정연,전영진,정구영 대한응급의학회 2001 대한응급의학회지 Vol.12 No.3

Background: In recent years, there has been increasing interest in the use of capnometry, the noninvasive, continuous measurement of end-tidal carbon dioxide(ETCO2) in expired air during CPR. The purpose of this study is to determine the significance of ETCO2 monitoring according to immediate cause of arrest during CPR as a prognostic indicator of successful resuscitation and survival. Methods: A prospective, clinical study was performed from May 1997 to December 2000 at the Department of Emergency Medicine, Ewha Womans University Mokdong Hospital. The study included 220 patients(231cases). All patients were immediately connected to a mainstream capnometer sensor between the tube and the bag after endotracheal intubation using an infrared capnometer. Results: The 107 patients(46.3%) with return of spontaneous circulation(ROSC) had higher maximal ETCO2 during CPR than the 113 patients without ROSC(31.0±19.4 vs 11.7±9.4 mmHg, P=0.000). The ETCO2 was not significantly different in relation to age, initial rhythm, and survival time after ROSC, but there was a significant difference in the immediate cause of arrest in the ROSC group(respiratory arrest: 40.2±23.5 mmHg, P=0.000). In case of cardiac arrest due to trauma, maximal ETCO2 was not significant in the ROSC group compared with the non-ROSC group(18.2±16.6 vs 10.8±7.5 mmHg, P=0.208). When maximal ETCO2 was less than 10 mmHg, we observed a sensitivity of 94.4% and a specificity of 39.5% in predicting ROSC. There were 6 patients with ROSC even though the maximal ETCO2 was less than 10 mmHg. Conclusion: Continuous ETCO2 monitoring during CPR may be noninvasive and valuable predictor of successful resuscitation and survival from cardiac arrest. However, ETCO2 should not be used as a single indicator for either cardiac arrest due to trauma or withdrawal of CPR.

정상인에서 전기 및 자기 자극을 이용한 설 근육의 전기생리학적 평가

조은미,류종현,안성환,조경원,임건한,김진호,김후원 朝鮮大學校 附設 醫學硏究所 2006 The Medical Journal of Chosun University Vol.31 No.1

Background: Abnormality of lingual muscles is clinically common. But its exact clinical assessment is quite difficult. Evaluation of motor power of the lingual muscles and their neural pathway is limited because of anatomical inaccessibility and individual variation of subjective symptoms. Transcranial magnetic stimulation (TMS) of the human motor cortex is a non-invasive tool to assess the functional integrity of the fast conducting central motor pathways and electrical stimulation of lingual nerve is a conventional method of peripheral nerve conduction. Therefore both study will be helpful to understanding of pathophysiology of lingual abnormality. Until now there is no such report in Korea. This study was conducted to acquire the easier method and normal control values of Korean subjects. Method: We recorded the motor responses from the lingual muscles of 32 healthy subjects (22 men, 10 women) by focal cortical TMS with a circular coil. Motor potentials were recorded by means of surface electrodes applied on the both side of the tongue, following TMS several centimeters lateral from the vertex and parietooccipital cortex and peripheral electrical stimulation of the 12th cranial nerve at the lateral 1/3 point of the line between the midline and angle of mandible. Results: During moderate motor activation, majority of maximal motor response were obtained 4-6cm lateral to the vertex with similar latencies. And the responses from magnetic stimulation of the intracranial segment of hypoglossal nerve were discarded because of difficulty to obtain and unsatisfactory reproducibility. The latency of motor evoked potentials in right cortical stimulation was 10.08 ± 1.17 msec and in left stimulation 10.04 ± 1.32 msec. The amplitudes in right 2.15 ± 1.02, left 2.09 ± 1.11 mV. The latencies electric hypoglossal nerve stimulation were 2.42 ± 0.34 in right, 2.49 ± 0.35 msec in left side. The amplitudes were 2.90 ± 2.37 in right, 2.43 ± 1.86 mV in left side. Central conduction times as calculated by subtracting the response latencies obtained by magnetic and electric stimulation are right 7.65 ± 1.19, left 7.55 ± 1.35 msec. There was no significant difference between man and woman, Conclusion: The method may be efficient, noninvasive, painless, and easy reproduced. With more extensive application to various neurological disorders, it comes close to being an ideal clinical conduction study technique for this cranial nerve.