• Asian-Australasian Journal of Animal Sciences
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• 제26권12호
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• pp.1680-1688
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• 2013

Many different approaches have been developed to improve the efficiency of animal cloning by somatic cell nuclear transfer (SCNT), one of which is to modify histone acetylation levels using histone deacetylase inhibitors (HDACi) such as trichostatin A (TSA). In the present study, we examined the effect of TSA on in vitro development of porcine embryos derived from SCNT. We found that TSA treatment (50 nM) for 24 h following oocyte activation improved blastocyst formation rates (to 22.0%) compared with 8.9% in the non-treatment group and total cell number of the blastocysts for determining embryo quality also increased significantly ($88.9{\rightarrow}114.4$). Changes in histone acetylation levels as a result of TSA treatment were examined using indirect immunofluorescence and confocal microscopy scanning. Results showed that the histone acetylation level in TSA-treated embryos was higher than that in controls at both acetylated histone H3 lysine 9 (AcH3K9) and acetylated histone H4 lysine 12 (AcH4K12). Next, we compared the expression patterns of seven genes (OCT4, ID1; the pluripotent genes, H19, NNAT, PEG1; the imprinting genes, cytokeratin 8 and 18; the trophoblast marker genes). The SCNT blastocysts both with and without TSA treatment showed lower levels of OCT4, ID1, cytokeratin 8 and 18 than those of the in vivo blastocysts. In the case of the imprinting genes H19 and NNAT, except PEG1, the SCNT blastocysts both with and without TSA treatment showed higher levels than those of the in vivo blastocysts. Although the gene expression patterns between cloned blastocysts and their in vivo counterparts were different regardless of TSA treatment, it appears that several genes in NT blastocysts after TSA treatment showed a slight tendency toward expression patterns of in vivo blastocysts. Our results suggest that TSA treatment may improve preimplantation porcine embryo development following SCNT.

• Asian Pacific Journal of Cancer Prevention
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• 제17권12호
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• pp.5057-5061
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• 2016

The present study was conducted to assess error rates with diagnosis using intra-operative frozen sections, and to indicate ways to increase overall performance. Over a period of two years, 227 cases were biopsied intra-operatively. Errors were observed in 14 cases. Four of these were sampling errors, one by a pathologist and three by surgeons. In seven cases incorrect interpretations were made. Epithelial dysplasia was observed on definitive histology in two cases which was not reported intra-operatively. One case was of ectopic thyroid. In cases of oral cancer where sentinel lymph nodes were sampled, immunohistochemistry for cytokeratin was performed to facilitate identification of micrometastasis. Only single case displayed tumor deposits which was not evident morphologically. Resection margins were reported in seventy eight cases. Some 18% (14/50) benefited from revision of margins; overall sensitivity of intra-operative frozen sections for marginal status was 71.4%, with a specificity of 90.3%. Overall sensitivity was 75% and specificity was 97.5%. Careful observation, pathologist experience and knowledge of limitations help in improving the overall diagnostic outcome.

• Animal Bioscience
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• 제36권1호
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• pp.43-52
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• 2023

Objective: This study aimed to examine the influence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection on pregnancy in cytokeratin-18 (K18)-hACE2 transgenic mice. Methods: To determine the expression of hACE2 mRNA in the female reproductive tract of K18-hACE2 mice, real-time polymerase chain reaction (RT-PCR) was performed using the ovary, oviduct, uterus, umbilical cord, and placenta. SARS-CoV-2 was inoculated intranasally (30 μL/mouse, 1×10⁴ TCID₅₀/mL) to plug-checked K18-hACE2 homozygous female mice at the pre-and post-implantation stages at 2.5 days post-coitum (dpc) and 15.5 dpc, respectively. The number of implantation sites was checked at 7.5 dpc, and the number of normally born pups was investigated at 20.5 dpc. Pregnancy outcomes, including implantation and childbirth, were confirmed by comparison with the non-infected group. Tissues of infected mice were collected at 7.5 dpc and 19.5 dpc to confirm the SARS-CoV-2 infection. The infection was identified by performing RT-PCR on the infected tissues and comparing them to the non-infected tissues. Results: hACE2 mRNA expression was confirmed in the female reproductive tract of the K18-hACE2 mice. Compared to the non-infected group, no significant difference in the number of implantation sites or normally born pups was found in the infected group. SARS-CoV-2 infection was detected in the lungs but not in the female reproductive system of infected K18-hACE2 mice. Conclusion: In K18-hACE2 mice, intranasal infection with SARS-CoV-2 did not induce implantation failure, preterm labor, or miscarriage. Although the viral infection was not detected in the uterus, placenta, or fetus, the infection of the lungs could induce problems in the reproductive system. However, lung infections were not related to pregnancy outcomes.

• IMMUNE NETWORK
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• 제7권3호
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• pp.117-123
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• 2007

Background: This study was designed to investigate the role of the hepatocyte growth factor (HGF) with regards to differentiation of somatic stem cells originating from the human umbilical cord blood (UCB) into hepatic lineage cells in vitro culture system. Methods: Mononuclear cells from UCB were cultured with and without HGF based on the fibroblast growth factor (FGF)-1, FGF-2, and stem cell factor. The cultured cells were confirmed by immunofluorescent staining analysis with albumin (ALB), cytokeratin-19 (CK-19), and proliferating cell nuclear antigen (PCNA) MoAb. ALB and CK-18 mRNA were also evaluated by reverse transcription-polymerase chain reaction. In order to observe changes in proliferating capacity with respect to the cultured period, CFSE with affinity to proliferating cells were tagged and later underwent flow cytometry. Results: In the HGF-treated group, cultured cells had a large oval shaped appearance with adherent, but easily detachable characteristics. In the HGF-non treated group, these cells were spindle-shaped with strong adherent characteristics. Expressions of ALB and CK-19 were evident in HGF-treated group compared to non-expression of those in to HGF-non treated group. Dual immunostaining analysis of the ALB producing cells showed presence of PCNA in their nuclei, and ALB and CK-18 mRNA were detected on the 21st day of cultured cells in the HGF-treated group. Conclusion: Our findings suggest that HGF has a pivotal role in differentiating somatic stem cells of human UCB into hepatic lineage cells in vitro.

• Tuberculosis and Respiratory Diseases
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• 제42권6호
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• pp.846-854
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• 1995

연구배경: Cyfra 21-1은 상피종양세포의 세포질에 존재하는 cytokeratin 19의 분절로서 상피종양세포의 파괴시 혈중내로 유리되므로 그 혈중 농도를 측정하여 종양표지자로 이용할 수 있는 것으로 알려져 있다. 이에 저자들은 폐암, 폐결핵, 기타폐질환 및 정상대조군 환자들의 혈청내 Cyfra 21-1, SCC 항원 및 CEA의 농도를 측정하여 폐암의 종양표지자로서 Cyfra 21-1과 SCC 항원 및 CEA의 진단적 효용성을 비교 관찰하고자 하였다. 또한 편평상피세포암에서 Cyfra 21-1과 평상펴셰포암의 특이 종양표지자로 알려진 SCC 항원과의 진단적 민감도와 특이도의 차이를 비교하고 그 병기 진행에 따른 Cyfra 21-1의 혈중농도의 증가 여부를 관찰하고자 하였다. 방법: 1992년 12월부터 1993년 6월까지 서울중앙병원에 입원하여 조직생검으로 초진단된 원발성 폐암 79예(편평상피세포암 41예, 선암 18예, 기타의 미분화 비소세포양 14예, 소세포암 6예)와 폐결핵 32예, 기타폐질환 23예, 정상대조군 23예를 대상으로 하였다. Cyfra 21-1과 ELSA2-CEA를 사용하였고, SCC 항원은 방사계수측정 kit인 ABBOTT SCC RIABEAD를 사용하였다. 결과: 1) Cyfra 21-1의 혈중농도는폐암군이 평균({\pm}표준편차) $18.38{\pm}3.65\;ng/mL$로서 비교군 $1.16{\pm}0.53\;ng/mL$보다 유의하게 높았다(p<0.0001). SCC 항원은 폐암군에서 $3.53{\pm}6.06\;ng/mL$로서 비교군 $1.19{\pm}0.5\;ng/mL$보다 유의하게 높았다(p<0.01). CEA는 폐암군에서 $35.03{\pm}13.9\;ng/mL$로서 비교군 $2.89{\pm}1.01\;ng/mL$ 보다 유의하게 높았다(p<0.0001). 2) 폐암군내에서는 Cyfra 21-1의 혈중농도가 편평상 피세포암군에서 $31.52{\pm}40.13\;ng/mL$로서 선암군 $2.41{\pm}1.34\;ng/mL$(p<0.0001) 및 소세포암군 $2.15{\pm}2.05\;ng/mL$(p=0.007) 보다 유의하게 높았다. SCC 항원의 혈중농도는 편평상피세포암군에서 $5.1{\pm}7.68\;ng/mL$로서 선암군 $1.36{\pm}0.69\;ng/mL$(p=0.009) 및 소세포암군 $1.1{\pm}0.24\;ng/mL$(p=0.024)보다 유의하게 높았다. 3) 편평상피세포암군에서 폐암의 병기 진행에 따른 Cyfra 21-1의 혈중농도의 증가는 없었다. 4) Cyfra 21-1의 진단양성 기준치를 3.3 ng/mL로 하였을때 편평상피세포암의 민감도가 83%로 선암의 22%, 소세포암의 17%보다 높게 산출되었다. SCC 항원의 민감도가 편평상피세포암에서 39%, 선암에서 11%, 소세포암에서 0% 이었다. CEA의 민감도가 편평상피세포암에서 20%, 선암에서 39%, 소세포암에서 33%이었다. 5) ROC 곡선 분석상 폐암의 진단에서 Cyfra 21-1의 민감도와 특이도가 SCC 항원 및 CEA 보다 우수한 것으로 나타났다. 결론: Cyfra 21-1은 폐임에서 SCC 항원 및 CEA에 비하여 민감도 및 특이도가 높은 종양표지자이며, 특히 편평상피세포암에서 그 민감도와 특이도가 높아 편평상피세포암의 특이 종양표지자로 알려진 SCC 항원보다 우수한 종양표지자로 사료되었다.

• Imaging Science in Dentistry
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• 제32권1호
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• pp.11-18
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• 2002

Purpose: To evaluate cultured human artificial skin as an experimental model for studying radiation effects in vitro. Materials and Methods: The skin was constructed by culturing keratinocytes over collagen lattice which made by culturing fibroblasts. Two groups were irradiated to gamma rays at single dose of 25 Gy with or without 3.5% of DMSO. Ultrastructures were investigated by electron microscopy after irradiation. The number of epidermal layers and expression of cytokeratin (CK) 14 & 10 were also seem by light microscopy. Results: At 2 days after irradiation in experimental group without DMSO, necrotic cells were rarely found in the spinosal layer and undercornified cells were visible in the homey layer. Similar findings were also found in experimental group with DMSO but in mild form. The number of epidermal layers in experimental group without DMSO were significantly fewer than other group. CK 14 expressed in all the layer excluding homey layer but CK 10 expressed over 3∼4 basal layers. Such patterns of CK expression were similar to all groups. It is suggested that structures of the keratinocytes and epidermal formation could be disturbed by irradiation in artificial skin and that DMSO can protect these damages. Conclusion : Therefore this work could be used as an organotypic experimental model in vitro using human cells for studying radiation effect in skin. Furthermore structural findings provided in this study could be used as useful basic data in further study using this model.

• 대한세포병리학회지
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• 제18권1호
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• pp.29-35
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• 2007

This study purposed to evaluate a $CellPrep^{(R)}$ (CP) of liquid-based cytology (LBC) to search for a less expensive and automated alternative cytologic preparation technique applicable to usually encountered cytologic specimens. Cervicovaginal direct-to-vial split samples from 457 gynecologic patients, 40 body fluid samples, and 34 urine samples were processed with the CP technique and the results were compared with those of currently used $ThinPrep^{(R)}$ (TP) method. Both CP and TP methods provide evenly distributed thin layers of cells with little cellular overlaps or significant obscuring elements in most of cases. Staining quality of both preparations showed a little difference due to the difference of fixative solutions without significant distractions in cytologic interpretation. On the supposition that TP was a gold standard, sensitivity, specificity, positive predictive value, and negative predictive value of CP cytology were 89%, 98%, 86%, and 99% in the cervicovaginal smear, 89%, 82%, 80%, and 90% in body fluid, and all of these values were 100% in urine samples. To testify the availability of immunohistochemistry on CP preparations, cytokeratin, vimentin, and Ki-67 were applied on body fluid specimens, and all of these antibodies were specifically stained on targeted cells. Conclusively, the CP method gave comparable results to those of TP in terms of smear quality and cytologic diagnostic evaluation, and was available on immunohistochemistry. The CP method could offer a cost-effective and automated alternative to the current expensive techniques of liquid-based cytology on popular cytologic materials including cervicovaginal, body fluid, and urine specimens.

• 핵의학기술
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• 제17권2호
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• pp.101-106
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• 2013

Purpose: As UBC IRMA is being tested, patients out of the reference value are reacting within the value again a few days later the urine collection tested, which causes the reliability of the test to decrease as a result. In this study, we can assume that the physiological changes in the factors would affect the results. The purpose of the study is to find out whether hematuria and proteinuria in the sample as well as the interval time (3 hours or more recommended) have affected the results. As a result, we could discover the changes in factors and increase the reliability of the test. Materials and Methods: 468 people (female: 249, male: 219) who came for the check-up were presented herein for medical examination from 2013.3.15 to 2013.1.2. Some people out of 468 who have reacted onto the reference value were divided into group low titer zone, ow-middle titer zone, and middle-high titer zone and tested for hematuria and proteinuria. During that period, 48 outpatients were asked to fill in a questionnaire regarding the urination interval time. The reagents used were (IDL Biotech AB, Sweden) and UBC IRMA. Results: Of the patients that are formed in the reference value of ($0.1-34.0{\mu}g/L$) turn out to be 52.7 years average age in their low concentration, ($mean{\pm}SD$) of the value of $0.10{\pm}0.02{\mu}g/L$. Among 80 people (50.8%, female: 49.2%), 16 patients (20%) have shown reaction to microscopic hematuria and 10 patients (12.5%) responded to proteinuria. In the average low concentration under 52.5 years of average age, 43 people (53%) have shown reaction to microscopic hematuria and 21 people (26.3%) are proteinuric patients out of 80 patients (male: 50.8%, female: 51.3%). In the middle high concentration of $11.8{\pm}4.82{\mu}g/L$ under the average age 51.7 years, 35 patients (53%) have responded to the microscopic hematuria and proteinuric patients are 26 people (39.3%) out of 66 people (men: 44%, women: 56%). In addition, in the concentration of $51.7{\pm}43.5{\mu}g/L$, some patients who get out of the reference value are observed as the average age of 52.0. 11 patients (78.6%) out of 14 (male: 35.7%, female: 64.3%) react to the microscopic hematuria. There show 6 people (42.8%) who turn out to be as proteinuric patients. As for the interval time, $1.67{\pm}3.71{\mu}g/L$ was the average value among 48 patients (female: 45.8%, male: 54.2%). Conclusion: We cannot see if proteinuria and hematuria directly affect abnormal results of inspection of 8,18 cytokeratin; however, we can find out that they statistically have an influence on highly generating UBC among several mechanisms. Also, although urination interval time was various every 15 minutes, we it does not affect these results.

• Tuberculosis and Respiratory Diseases
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• 제44권4호
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• pp.785-795
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• 1997

연구배경 : 종양 표지자는 각종 악성 종양의 진단, 진행정도, 절제 가능성, 예후추정, 치료후 추적 검사 등에도 사용될 수 있으나 민감도와 특이도가 만족스럽지 못한 경우가 많아서 임상에서 보조적인 수단으로 이용되고 있는 경우가 대부분이다. 최근 Cyfra21-1이 비소세포 폐암의 진단에 CEA, SCC보다 우수하다는 보고가 있었고 단일클론 항체를 이용한 TPA-M(Tissue polypeptide antigen)이 비소세포 폐암은 진단하는데 상당히 우수한 결과를 보인다고 보고하고 있다. 이에 폐암환자에서 혈청 CEA, SCC, Cyfra21-1, TPA-M의 임상적 유용성을 서로 비교해 보고자 이 연구를 시행하였다. 방 법 : 1996년 4월 1일 부터 19961건 8월 31일까지 계명대학교 의과대학 병원에 내원한 환자중 조직 생검으로 확진된 비소세포 폐암 37예와 소세포 폐암 12예를 총 49예를 폐암 환자군으로 같은 기간의 입원 환자 중 양성 양성 폐질환으로 확진된 29예를 대조군으로 선정하였고 폐암 환자의 경우 수술, 항암요법 및 방사선치료를 전혀 시행하지 않은 상태에서 혈청을 채취하여 검사를 시행하였다. CEA는 일본 Eiken사의 Ab bead CEA, SCC는 미국 DAINABOT사의 SCC RIA BEAD kit, Cyfra21-1은 일본 TFB사의 CA 21-1 kit, TPA-M은 일본 DAIICHI사의 TPA-M kit를 사용하였다. 결 과 : 전체 폐암환자와 대조군을 비교해보면 CEA가 $10.05{\pm}38.39{\mu}/L$, $1.59{\pm}0.94{\mu}/L$, SCC가 $3.04{\pm}5.79{\mu}/L$, $1.58{\pm}2.85{\mu}/L$, Cyfra21-1이 $8.27{\pm}11.96{\mu}/L$, $1.77{\pm}2.72{\mu}/L$, TPA-M이 $132.02{\pm}209.35U/L$, $45.86{\pm}75.86U/L$으로 나타났으며 Cyfra21-1과 TPA-M은 폐암환자에서 대조군보다 종양 표지자의 측정치가 유의하게 높았다.(p<0.05) CEA, Cyfra21-1, TPA-M, SCC의 cutoff value를 회사에서 권장하는 $2.5{\mu}/L$, $3.0{\mu}/L$, 70.0U/L, $2.0{\mu}/L$로 하였을 때 전체폐암에 대한 민감도와 특이도는 CEA 33.3%와 78.6%, Cyfra21-1 50.0%와 89.7%, TPA-M 52.3%와 89.7%, SCC 23.8%와 89.3%이었으며 비소세포 폐암에 대하여서는 CEA 36.1%와 78.1%, Cyfra21-1 50.1%와 89.7%, TPA-M 53.1%와 89.7%, SCC 33.8%와 89.3% 소세포 폐암의 경우에도 CEA 25.0%와 78.5%, Cyfra21-1 50.0%와 89.6%, TPA-M 50.0%와 89.6%, SCC 0%와 89.2%로 폐암환자에서 Cyfra21-1과 TPA-M이 민감도와 특이도에 있어 더 우수한 결과를 보였고 비소세포 폐암환자에서 SCC도 높은 특이도를 나타내었다. ROC(Receiver operating characteristic) curve에 따라 민감도와 특이도를 얻어 가장 정확도가 높은 cutoff value를 CEA $1.25{\mu}/L$, Cyfra21-1 $1.5{\mu}/L$, TPA-M 35U/L, SCC $0.6{\mu}/L$로 하여 민감도와 특이도, 정확도 및 kappa index를 비교해 보아도 Cyfra21-1과 TPA-M이 더 우수한 결과를 나타내었다. 병기에 따른 비소세포 폐암에서의 종양 표지자를 3A 병기까지와 3B 병기이상으로 나누어 비교해보면 3B 병기이상에서 SCC에서만 통계적 유의성을 보였다. (p<0.05) 그러나 종양크기에 따른 종양표지자의 측정치는 통제적 유의성이 없었다.(p>0.05) 결 론 : 혈청 TPA-M과 Cyfra21-1은 폐암환자에 있어 CEA나 SCC보다 민감도와 특이도가 높으며 SCC의 경우 비소세포 폐암에 있어 높은 특이도를 보였으며 SCC의 측정치가 병기와 유관하게 나타났으나 종양의 크기와 병기의 진행과의 관계에 있어서는 더 많은 환자들을 대상으로 한 연구가 행해져야 할 것으로 생각된다.