Oral bacterial infections substantially affect the development of various periodontal diseases and oral cancers. However, the molecular mechanisms underlying the association between Fusobacterium nucleatum (F. nucleatum ), a major periodontitis (PT)-associated pathogen, and these diseases require extensive research. Previously, our RNAsequencing analysis identified a few hundred differentially expressed genes in patients with PT and peri-implantitis (PI) than in healthy controls. Thus, in the present study using oral squamous cell carcinoma (OSCC) cells, we aimed to evaluate the effect of F. nucleatum infection on genes that are differentially regulated in patients with PT and PI. Human oral squamous cell carcinoma cell lines OSC-2O, HSC-4, and HN22 were used. These cells were infected with F. nucleatum at a multiplicity of infection of 100 for 3 hours at 37℃ in 5% CO2. Gene expression was then measured using reverse-transcription polymerase chain reaction. Among 18 genes tested, the expression of CSF3, an inflammation-related cytokine, was increased by F. nucleatum infection. Additionally, F. nucleatum infection increased the phosphorylation of AKT, p38 MAPK, and JNK in OSC-20 cells. Treatment with p38 MAPK (SB202190) and JNK (SP600125) inhibitors reduced the enhanced CSF3 expression induced by F. nucleatum infection. Overall, this study demonstrated that F. nucleatum promotes CSF3 expression in OSCC cells through p38 MAPK and JNK signaling pathways, suggesting that p38 MAPK and JNK inhibitors may help treat F. nucleatum-related periodontal diseases by suppressing CSF3 expression.
본 연구의 목적은 60대의 대뇌에서 T1, T2, PD 이완 시간 값을 측정하여 연령별 특정 해부학적 구조물들의 이완 시간 의 평균값과 연령에 따른 이완 시간 변화의 관련성이 있는지 분석하고자 하였다. 60에서 69세까지 총 50명의 정상 뇌자기 공명영상검사자의 데이터를 Synthetic MR의 MAGiC을 이용하여 후향적으로 분석하였다. 대상 부위는 해마, 대뇌 부챗살, 측두엽 회백질, 시상, 뇌척수액이었다. 실험결과 해마, 대뇌 부챗살, 측두엽 회백질은 연령 변화에 따른 T1, T2, PD 이완 시간의 차이가 없었다(p>0.05). 하지만 시상에서는 PD 이완시간이 연령과의 상관성이 있었고(R2=0.112, p<0.05), 뇌척 수액에서는 T1 이완시간(R2=0.063, p<0.05)과 T2 이완시간(R2=0.061, p<0.05)에서 연령과의 상관성을 확인하였다. 추후 다양한 연령대의 대뇌 이완 시간을 측정하여 평균값의 비교 연구가 필요하고, 시상과 뇌척수액에서는 대규모 모집단 연구가 필요할 것이라 판단되며 이에 본 연구가 기초자료를 제공할 것이라 사료된다.
The purpose of this study was to analyze trends in sustainable fashion research and to guide its the development by examining previous research projects conducted by the UK Centre for Sustainable Fashion. A literature review and case studies were conducted from which the following results were identified. It was shown that the CSF has cooperated with fashion, arts and technology businesses and other external organizations. The CSF has contributed to improving the economic durability of the fashion industry, the convergence of both local and the international fashion communities, and the development of cultural sustainability. Moreover, it incorporates various academic fields into its research and has developed advanced researching methods for sustainable fashion. The CSF has embraced the new perspectives by utilizing student ideas for the development of sustainable fashion. The conclusions of this study are as follows. First, the CSF studied the sustainability of fashion from a more organic perspective through multidisciplinary research. Second, it has expanded the possibility of sustainable fashion research through research formats, research fields and research operations. Finally, it has evoked values and responsibilities that the fashion industry should pursue in modern society and has become an exemplary case of sustainable fashion research, providing creative studies for sustainable fashion research in the future.
The clustered regularly interspaced short plalindromic repeats(CRISPR)/CRISPR associated protein (Cas9) system can be applied to produce transgenic pigs. We applied CRISPR/Cas9 system to generate hG-CSF targeted pig parthenogenetic embryos. Using sigle guided RNA targeted to pig hG-CSF genes was injected into cytoplasm of in vitro matured oocyte before electrical activation. The CRISPR/Cas9 vector were diluted in Tris-EDTA buffer (TE buffer) and injected with different concentration of 0 (sham injection), 2.5 and 25 ng/ul. In results, regardless of the concentrations of vector, the cleavage and blastocyst rate were not significantly different among three groups. Since plasmid DNA was used for microinjection, we investigated whether DNA vectors were integrated into the genome. Genomic PCR of the coding sequence of Cas9 variants and hG-CSF was performed to detect genomic integrants. Each blastocysts were collected into a microtube, and then PCR was performed. Overall 32 embryos are not expressed targeted gene.
The majority of early embryonic mortality in pregnancy occurs during the peri-implantation stage, suggesting that this period is important for conceptus viability and the establishment of pregnancy. Successful establishment of preg-nancy in all mammalian species depends on the orchestrated molecular events that transpire at the conceptus- uterine interface during the peri-implantation period. This maternal-conceptus interaction is especially crucial in pigs because in them non-invasive epitheliochorial placentation occurs, in which the pre-implantation phase is prolonged. During the pre-implantation period, conceptus survival and the establishment of pregnancy are known to depend on the developing conceptus receiving an adequate supply of histotroph, which contains a wide range of nutrients and grow-th factors. Evidence links growth factors including epidermal growth factor (EGF), insulin-like growth factor-I (IGF-I), vascular endothelial growth factor (VEGF), and colony-stimulating factor 2 (CSF2) to embryogenesis or implantation in various mammalian species; however, in the case of pig, little is known about such functions of these growth factors, especially their regulatory mechanisms at the maternal-conceptus interface. Our research group has presented evidence for promising growth factors affecting cellular activities of primary porcine trophectoderm (pTr) cells, and we have identified potential intracellular signaling pathways responsible for the activities induced by these factors. Therefore, this review focuses on promising growth factors at the maternal-conceptus interface regulating the develop-ment of the porcine conceptus and playing pivotal roles in implantation events during early pregnancy in pigs.
본 연구에서는 hG-CSF의 발현을 유도적으로 조절하기 위한 FIV-Tet-On lentivirus vector system을 구축하고자 하였다. hG-CSF는 호중성구 계열 세포의 증식과 분화, 생존에 영향을 미치는 물질로서, 이 유전자의 발현을 증가시키기 위하여 FIV-Tet-On vector 상의 hG-CSF나 rtTA2SM2 서열의 3' 위치에 WPRE 서열을 도입하였다. 구축된 각각의 vector는 293FT 세포에 일시적으로 transfection하여 virus를 생산하였으며, 이 virus를 일차 배양 세포인 CEF와 PFF에 감염시켰다. 각 세포에 전이된 hG-CSF의 발현 양상을 관찰하기 위하여 doxycycline을 첨가하거나 첨가하지 않은 배지에서 배양한 후 quantitative real-time PCR, Western blot과 ELISA를 이용하여 hG-CSF 유전자의 발현 정도를 비교 측정한 결과, CEF에서는 WPRE가 hG-CSF의 3' 위치에 도입된 경우에 발현량과 유도율이 가장 높은 것으로 나타났고, PFF에서는 rtTA 서열의 3'위치에 도입된 경우에 발현량과 유도율이 가장 큰 것으로 확인되었다. 이 FIV-Tet-On vector system은 형질 전환 동물의 생산이나 유전자 치료에서 문제시되는 외래 유전자의 지속적인 과다 발현에 의한 개체의 생리적인 부작용을 최소화하기 위한 해결 방법으로 제시될 수 있을 것이다.