• Journal of Veterinary Science
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• 제25권1호
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• pp.1.1-1.15
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• 2024

Background: Axitinib, a potent and selective inhibitor of vascular endothelial growth factor (VEGF) receptor (VEGFR) tyrosine kinase 1,2 and 3, is used in chemotherapy because it inhibits tumor angiogenesis by blocking the VEGF/VEGFR pathway. In veterinary medicine, attempts have been made to apply tyrosine kinase inhibitors with anti-angiogenic effects to tumor patients, but there are no studies on axitinib in canine mammary gland tumors (MGTs). Objectives: This study aimed to confirm the antitumor activity of axitinib in canine mammary gland cell lines. Methods: We treated canine MGT cell lines (CIPp and CIPm) with axitinib and conducted CCK, wound healing, apoptosis, and cell cycle assays. Additionally, we evaluated the expression levels of angiogenesis-associated factors, including VEGFs, PDGF-A, FGF-2, and TGF-β1, using quantitative real-time polymerase chain reaction. Furthermore, we collected canine peripheral blood mononuclear cells (PBMCs), activated them with concanavalin A (ConA) and lipopolysaccharide (LPS), and then treated them with axitinib to investigate changes in viability. Results: When axitinib was administered to CIPp and CIPm, cell viability significantly decreased at 24, 48, and 72 h (p < 0.001), and migration was markedly reduced (6 h, p < 0.05; 12 h, p < 0.005). The apoptosis rate significantly increased (p < 0.01), and the G2/M phase ratio showed a significant increase (p < 0.001). Additionally, there was no significant change in the viability of canine PBMCs treated with LPS and ConA. Conclusion: In this study, we confirmed the antitumor activity of axitinib against canine MGT cell lines. Accordingly, we suggest that axitinib can be applied as a new treatment for patients with canine MGTs.

• Animal Bioscience
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• 제37권1호
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• pp.50-60
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• 2024

Objective: Testicular fat deposition has been reported to affect animal reproduction. However, the underlying mechanism remains poorly understood. The present study explored whether sperm meiosis and testosterone synthesis contribute to mouse testicular fat deposition-induced reproductive performance. Methods: High fat diet (HFD)-induced obesity CD1 mice (DIO) were used as a testicular fat deposition model. The serum hormone test was performed by agent kit. The quality of sperm was assessed using a Sperm Class Analyzer. Testicular tissue morphology was analyzed by histochemical methods. The expression of spermatocyte marker molecules was monitored by an immuno-fluorescence microscope during meiosis. Analysis of the synthesis of testosterone was performed by real-time polymerase chain reaction and reagent kit. Results: It was found that there was a significant increase in body weight among DIO mice, however, the food intake showed no difference compared to control mice fed a normal diet (CTR). The number of offspring in DIO mice decreased, but there was no significant difference from the CTR group. The levels of follicle-stimulating hormone were lower in DIO mice and their luteinizing hormone levels were similar. The results showed a remarkable decrease in sperm density and motility among DIO mice. We also found that fat accumulation affected the meiosis process, mainly reflected in the cross-exchange of homologous chromosomes. In addition, overweight increased fat deposition in the testis and reduced the expression of testosterone synthesis-related enzymes, thereby affecting the synthesis and secretion of testosterone by testicular Leydig cells. Conclusion: Fat accumulation in the testes causes testicular cell dysfunction, which affects testosterone hormone synthesis and ultimately affects sperm formation.

• 대한화장품학회지
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• 제50권1호
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• pp.29-36
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• 2024

본 연구는 RAW 264.7 세포를 이용하여 넓패추출물(Ishige foliacea, I. foliacea)의 피부 항염 효과를 확인하고자 하였다. RAW 264.7 세포에 대한 넓패추출물의 항염증 효과는 세포 생존력 분석, mRNA 발현 및 산화질소(NO)/프로스타글란딘 E₂ (PGE₂) 생성을 통해 확인하였다. 넓패추출물의 항염증 효과는 IL-1α/IL-1β/IL-6/TNFα mRNA 발현 및 NO/PGE₂ 생성을 분석하여 규명하였고, 정량적 실시간 중합효소 연쇄반응을 통해 넓패추출물은 iNOS/COX2/IL-1α/IL-1β 및 IL-6의 mRNA 발현을 감소시키는 것을 확인하였다. 또한 PGE₂와 NO 생성의 비교를 통해 넓패추출물이 항염증 효과가 있음을 확인하였다. 넓패추출물의 항염 효과를 바탕으로 코스메슈티컬 소재로 활용될 수 있는 가능성을 확인하였다. 넓패추출물의 항염증 메커니즘에 대한 후속 연구를 바탕으로 코스메슈티컬뿐만 아니라 염증과 관련된 약리학적인 메커니즘 발굴에도 도움이 될 것으로 보인다.

• Korean Journal of Radiology
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• 제21권10호
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• pp.1150-1160
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• 2020

Objective: To describe the experience of implementing a deep learning-based computer-aided detection (CAD) system for the interpretation of chest X-ray radiographs (CXR) of suspected coronavirus disease (COVID-19) patients and investigate the diagnostic performance of CXR interpretation with CAD assistance. Materials and Methods: In this single-center retrospective study, initial CXR of patients with suspected or confirmed COVID-19 were investigated. A commercialized deep learning-based CAD system that can identify various abnormalities on CXR was implemented for the interpretation of CXR in daily practice. The diagnostic performance of radiologists with CAD assistance were evaluated based on two different reference standards: 1) real-time reverse transcriptase-polymerase chain reaction (rRT-PCR) results for COVID-19 and 2) pulmonary abnormality suggesting pneumonia on chest CT. The turnaround times (TATs) of radiology reports for CXR and rRT-PCR results were also evaluated. Results: Among 332 patients (male:female, 173:159; mean age, 57 years) with available rRT-PCR results, 16 patients (4.8%) were diagnosed with COVID-19. Using CXR, radiologists with CAD assistance identified rRT-PCR positive COVID-19 patients with sensitivity and specificity of 68.8% and 66.7%, respectively. Among 119 patients (male:female, 75:44; mean age, 69 years) with available chest CTs, radiologists assisted by CAD reported pneumonia on CXR with a sensitivity of 81.5% and a specificity of 72.3%. The TATs of CXR reports were significantly shorter than those of rRT-PCR results (median 51 vs. 507 minutes; p < 0.001). Conclusion: Radiologists with CAD assistance could identify patients with rRT-PCR-positive COVID-19 or pneumonia on CXR with a reasonably acceptable performance. In patients suspected with COVID-19, CXR had much faster TATs than rRT-PCRs.

• Journal of Animal Science and Technology
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• 제66권1호
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• pp.204-218
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• 2024

Elsholtzia fruticosa (EF) is present in tropical regions throughout South Asian countries as well as the Himalayas. Although it has been used as a traditional medicine to treat digestive, respiratory, and inflammatory issues, its effect on preadipocyte differentiation is unknown. In this study, we examined the effects of a methanol extract prepared from EF on the differentiation of 3T3-L1 preadipocytes. Cell differentiation was assessed by microscopic observation and oil-red O staining. The expression of adipogenic and lipogenic genes, including PPARγ and C/EBPα, was measured by western blot analysis and quantitative real-time polymerase chain reaction (qRT-PCR), to provide insight into adipogenesis and lipogenesis mechanisms. The results indicated that EF promotes the differentiation of 3T3-L1 preadipocytes, with elevated lipid accumulation occurring in a concentration-dependent manner without apparent cytotoxicity. EF enhances the expression of adipogenic and lipogenic genes, including PPARγ, FABP4, adiponectin, and FAS, at the mRNA and protein levels. The effect of EF was more pronounced during the early and middle stages of 3T3-L1 cell differentiation. Treatment with EF decreased C/EBP homologous protein (CHOP) mRNA and protein levels, while increasing C/EBPα and PPARγ expression. Treatment with EF resulted in the upregulation of cyclin E and CDK2 gene expression within 24 h, followed by a decrease at 48 h, demonstrating the early-stage impact of EF. A concomitant increase in cyclin-D1 levels was observed compared with untreated cells, indicating that EF modulates lipogenic and adipogenic genes through intricate mechanisms involving CHOP and cell cycle pathways. In summary, EF induces the differentiation of 3T3-L1 preadipocytes by increasing the expression of adipogenic and lipogenic genes, possibly through CHOP and cell cycle-dependent mechanisms.

• Animal Bioscience
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• 제37권7호
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• pp.1289-1302
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• 2024

Objective: There is a strong relationship between the content of beneficial fatty acids in milk and milk fat metabolic activity in the mammary gland. To improve milk quality, it is therefore necessary to study fatty acid metabolism in bovine mammary gland tissue. In adipose tissue, peroxisome proliferator-activated receptor gamma (PPARG), the core transcription factor, regulates the fatty acid metabolism gene network and determines fatty acid deposition. However, its regulatory effects on mammary gland fatty acid metabolism during lactation have rarely been reported. Methods: Transcriptome sequencing was performed during the prelactation period and the peak lactation period to examine mRNA expression. The significant upregulation of PPARG drew our attention and led us to conduct further research. Results: According to bioinformatics prediction, dual-luciferase reporter system detection, real-time quantitative reverse transcription polymerase chain reaction and Western blotting, miR-130a and miR-130b could directly target PPARG and inhibit its expression. Furthermore, triglyceride and oil red O staining proved that miR-130a and miR-130b inhibited milk fat metabolism in bovine mammary epithelial cells (BMECs), while PPARG promoted this metabolism. In addition, we also found that the coexpression of miR-130a and miR-130b significantly enhanced their ability to regulate milk fat metabolism. Conclusion: In conclusion, our findings indicated that miR-130a and miR-130b could target and repress PPARG and that they also have a functional superposition effect. miR-130a and miR-130b seem to synergistically regulate lipid catabolism via the control of PPARG in BMECs. In the long-term, these findings might be helpful in developing practical means to improve high-quality milk.

• Animal Bioscience
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• 제37권8호
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• pp.1367-1376
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• 2024

Objective: Parathyroid hormone like hormone (PTHLH), as an essential factor for bone growth, is involved in a variety of physiological processes. The aim of this study was to explore the role of PTHLH gene in the growth of antlers. Methods: The coding sequence (CDS) of PTHLH gene cDNA was obtained by cloning in sika deer (Cervus nippon), and the bioinformatics was analyzed. The quantitative real-time polymerase chain reaction (qRT-PCR) was used to analyze the differences expression of PTHLH mRNA in different tissues of the antler tip at different growth periods (early period, EP; middle period, MP; late period, LP). Results: The CDS of PTHLH gene was 534 bp in length and encoded 177 amino acids. Predictive analysis results revealed that the PTHLH protein was a hydrophilic protein without transmembrane structure, with its secondary structure consisting mainly of random coil. The PTHLH protein of sika deer had the identity of 98.31%, 96.82%, 96.05%, and 94.92% with Cervus canadensis, Bos mutus, Oryx dammah and Budorcas taxicolor, which were highly conserved among the artiodactyls. The qRT-PCR results showed that PTHLH mRNA had a unique spatio-temporal expression pattern in antlers. In the dermis, precartilage, and cartilage tissues, the expression of PTHLH mRNA was extremely significantly higher in MP than in EP, LP (p<0.01). In the mesenchyme tissue, the expression of PTHLH mRNA in MP was significantly higher than that of EP (p<0.05), but extremely significantly lower than that of LP (p<0.01). The expression of PTHLH mRNA in antler tip tissues at all growth periods had approximately the same trend, that is, from distal to basal, it was first downregulated from the dermis to the mesenchyme and then continuously up-regulated to the cartilage tissue. Conclusion: PTHLH gene may promote the rapid growth of antler mainly through its extensive regulatory effect on the antler tip tissue.

• Animal Bioscience
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• 제37권8호
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• pp.1377-1386
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• 2024

Objective: Embryonic interferon-tau (IFNT) and progesterone affect expression of interferon-stimulated genes (ISGs), progesterone receptor (PGR) and progesterone-induced blocking factor (PIBF) in the ovine thyroid. Methods: Thyroids of ewes were sampled at day 16 of nonpregnancy, days 13, 16, and 25 of pregnancy, and real-time quantitative polymerase chain reaction assay, western blot and immunohistochemistry were used to detect expression of ISGs, PGR, and PIBF. Results: Free ISG15 protein was undetected, but ISG15 conjugated proteins upregulated at day 16 of pregnancy, and expression levels of ISG15 conjugated proteins, PGR isoform (70 kDa), PIBF, interferon-gamma-inducible protein 10 and myxovirusresistance protein 1 peaked, but expression level of signal transducer and activator of transcription 1 was the lowest at day 16 of pregnancy. In addition, the expression levels of PGR isoform (70 kDa) and signal transducer and activator of transcription 1 (STAT1) decreased, but levels of PGR isoform (43 kDa), 2',5'-oligoadenylate synthetase, IP-10 and MX1 increased at day 25 of pregnancy comparing with day 16 of the estrous cycle. Conclusion: Early pregnancy affects expression of ISGs, PGR, and PIBF in maternal thyroid through IFNT and progesterone, which may regulate thyroid autoimmunity and thyroid hormone secretion in ewes.

• 대한소아치과학회지
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• 제45권3호
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• pp.271-279
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• 2018

소아에서 사이클로스포린에 의해 유발되는 치은과증식의 병인을 규명하기 위한 이전의 연구에서는, 개체 간의 차이를 유발할 수 있는 요인을 완전히 배제하지 않았다. 본 연구는 개체 별로 생물학적 변이의 영향을 통제한 실험 모델에 의해 사이클로스포린이 1형 교원질의 대사에 미치는 영향을 알아보았다. 5주령의 수컷 Sprague-Dawley rats 5마리에 6주 동안 사이클로스포린을 위장으로 투여하였다. 사이클로스포린 투여 전과 투여 후 2, 4, 6주째에 쥐의 하악구치부에서 치은을 채취하였다. 20개의 치은표본으로부터 치은섬유모 세포를 추출하였다. 개체 별로 사이클로스포린을 투여하기 전 채취한 치은섬유모세포의 반은 대조군, 나머지 반은 실험실 내 약물 처리한 실험군(Tt)으로 하였다. 사이클로스포린 투여 후 2, 4, 6주째 채취한 치은섬유모세포는 각각의 실험군으로서 T2, T4, T6으로 명명하였다. 면역형광법을 시행하여 모든 치은모세포 내의 1형 교원질의 존재를 확인하였다. 각 개체 별로 대조군과 실험군에서 1형 교원질의 유전자와 단백질 발현의 변화를 분석하기 위해서 각각 Real-time polymerase chain reaction과 Western blotting을 시행하였다. 사이클로스포린을 처리하기 전과 후를 한 개체내에서 비교하여 1형 교원질의 변화를 평가하였다. 대조군과 실험군에서 1형 교원질의 유전자 발현에서는 유의한 차이가 없었다. 모든 개체에서 실험실 내 사이클로스포린 처리한 치은섬유모세포의 1형 교원질의 단백질 발현이 증가한 경우, 사이클로스포린을 투여한 후 채취한 치은섬유모세포에서도 1형 교원질의 단백질 발현이 증가하였다. 본 연구에서 시행한 개체별로 발생할 수 있는 생물학적 변이를 모두 통제한 실험 방법은 유용하였다. 1형 교원질 이외에 다른 요인들에 대한 후속 연구 및 인간에서의 심도 있는 연구가 필요하다.

• 대한임상검사과학회지
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• 제49권1호
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• pp.28-39
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• 2017

잔류성 유기 오염 물질은 후성학적 메커니즘과 비만의 발달에 영향을 줄 수가 있다. 폴리브롬화 디페닐 에테르는 주요한 잔류성 유기 오염 물질 중 하나이며, 난연제로 널리 쓰인다. 출생전 잔류성 유기 오염 물질과 같은 내분비교란물질에 노출시 LINE-1 (long interspersed nuclear elements)의 global DNA 메틸화와 비만 위험도의 증가에 영향을 미칠 수 있다. 따라서, 이번 연구는 임신한 스프라그-돌리 백서를 이용하여 태반과 모유를 통하여 전달된 BDE-47, BDE-209가 LINE-1에서의 후성학적인 변화와 obesogen으로서 발달과정에 따른 유전적 비만 감수성의 증가에 영향을 줄 수 있는지에 대해서 보고자 하였다. 어미와 새끼에서 LINE-1의 광범위 DNA 메틸화와 비만과 관련된 유전자 발현은 methylation-sensitive high resolution melting analysis (MS-HRM), direct bisulfite sequencing와 quantitative real time polymerase chain reaction (qPCR)을 사용하여 각각 분석하였다. MS-HRM 결과는 출생 후 4일의 노출군 새끼에서 (4마리 중 2마리) LINE-1의 광범위 DNA 저메틸화 양상을 보여주었지만, bisulfite sequencing은 노출군과 비노출군에서 차이가 없었다. ${\beta}$-산화 경로와 adipokines과 관련된 어미의 유전자 발현은 두 그룹간 차이를 보였다. 반면에, 새끼의 유전자 발현은 비슷한 양상을 나타내었다. ${\beta}$-산화 경로와 비만과 관련된 유전자 발현 중 $PPAR-{\alpha}$를 제외하고는 출생 시에 유의하게 증가하였다. 결론적으로, 이번 연구는 BDE-47, BDE-209의 동시 노출이 태반과 모유를 통해서 새끼에서의 후성학적인 변화와 비만과 관련된 유전자 발현 변화에 영향을 미칠 수 있는 것을 보여주었다.