Oral squamous cell carcinoma (OSCC) is the most common type of head and neck cancer and is associated with high recurrence, poor treatment, and low survival rates. Hypoxia-inducible factor-1α (HIF-1α) is a transcription factor that regulates the response to hypoxia, a major factor in the tumor microenvironment that affects tumor development and progression in various cancer types. However, microRNA (miRNA) sequence analysis revealed that only a few miRNAs targeting HIF-1α had been discovered. In the present study, we investigated HIF-1α expression in OSCC and the effect of HIF-1α-targeting miRNAs on the progression and metastatic potential of OSCC. We analyzed public databases to explore which miRNAs target HIF-1α expression. In addition, the expression of proteins involved in the cell cycle, proliferation, and apoptosis in HSC-2 cells was analyzed after miRNA-126 mimic treatment. Furthermore, to investigate the effect of miRNA-126 on the proliferation and invasion ability of OSCC cells, 5-ethynyl-2′-deoxyuridine and Transwell assays were performed. The activities of MMP-2 and MMP-9 were evaluated via gelatin zymography. Our results showed that miRNA-126, which targets HIF-1α, enhances OSCC cell proliferation by regulating the cell cycle and reinforces the cell mobility of OSCC via HIF-1α expression. These findings suggest that miRNA-126 may be a novel marker for OSCC treatment and the development of new tools for patients with OSCC.

Inflammation is a protective mechanism against pathogens, but if maintained continuously, it destroys tissue structures. Aggregatibacter actinomycetemcomitans is a gram-negative, facultative anaerobic bacterium often found in severe periodontitis. A. actinomycetemcomitans invades epithelial cells and triggers inflammatory response in the immune cells. In this study, we investigated the effect of water-soluble rosehip extract on A. actinomycetemcomitansinduced inflammatory responses. A human monocytic cell line (THP-1) was differentiated to macrophages by phorbol 12-mystristate 13-acetate treatment. The cytotoxic effect of extract was determined using the 3-(4,5-dimethylthiazol-2- yl)-2,5-diphenyltetrazolium bromide assay. The effects of extract on bacterial growth were examined by measuring the optical densities using a spectrophotometer. THP-1-derived macrophages were infected A. actinomycetemcomitans after extract treatment, and culture supernatants were analyzed for cytokine production using enzyme-linked immunosorbent assay. Protein expression was measured by western blotting. Extract was not toxic to THP-1- derived macrophages. A. actinomycetemcomitans growth was inhibited by 1% extract. The extract suppressed A. actinomycetemcomitans-induced tumor necrosis factor-α, interleukin (IL)-1β, and IL-8 production. It also decreased mitogen-activated protein kinase (MAP kinase) and nuclear factor-κB (NF-κB) phosphorylation. Moreover, the extract inhibited the expression of inflammasome components, including nucleotide-binding oligomerization domain-like receptor pyrin domain-containing protein 3, Absent in Melanoma 2, and apoptosis associated speck-like protein containing a CARD. And cysteine-aspartic proteases-1 and IL-1β expression were decreased by the extract. In summary, extract suppressed A. actinomycetemcomitans growth and decreased inflammatory cytokine production by inhibiting activation of MAP kinase, NF-κB, and inflammasome signaling. Rosehip extract could be effective in the treatment of periodontal inflammation induced by A. actinomycetemcomitans infection.

We present the results of near-infrared imaging observations of the galaxy overdensity around the z = 1.44 radio-loud active galactic nucleus (AGN) 6CE1100+3505, which was carried out with the purpose of sampling the redshifted Hα emission from the actively star-forming galaxies that could constitute the overdensity. The existence of the structure around this AGN was spectroscopically confirmed by previous grism observations which are however limited to the central region. Using the CH4Off narrow/medium-band and H broad band filters in the Wide Infrared Camera (WIRCam) on the Canada-France-Hawaii Telescope (CFHT), we constructed a sample of objects that show a flux excess in the CH4Off band due to line emission. The emission line flux is ∼ 4.9 × 10−16 erg s−1 cm−2 , corresponding to a star formation rate (SFR) of ∼ 50 M⊙ yr−1 for galaxies at redshifts z ∼ 1.4. None of the galaxies with medium-band flux excess is located within 1 Mpc from the central AGN, and there is no evidence that the selected galaxies are associated with the proposed cluster. Along with the star formation quenching near the center that was found from the previous grism observations, the lack of extreme starbursts in the structure suggests that at z ∼ 1.4, overdense regions are no longer favorable locations for vigorous star formation.

정신분열은 대표적인 신경정신성 질환으로 정신분열 환자들은 정신적, 감정적 및 행동적인 장애로 인해 고통받고 있다. 이 연구는 흰쥐를 대상으로 정신분열을 유도함으로써 골격근 조직에서 반응하는 혈관생성 관련 단백질의 변화를 살펴보고 규칙적인 운동의 효과를 알아보고자 수행되었다. 연구의 목적을 위해 흰쥐(SD rats) 총 18마리를 통제(n=6), 정신분열(n=6), 정신분열+운동(n=6) 집단으로 구분하였다. 정신분열 및 정신분열+운동 집단은 총 2주 동안 MK-801처치를 통해 정신분열을 유도하였으며, 정신분열+운동 집단은 4주간 규칙적인 유산소성 운동을 수행하였다. 유산소성 운동은 실험동물용 트레드밀을 이용하여 수행되었으며 HIF-1α 및 VEGF 단백질은 western blot을 통해 관찰하였다. 연구결과, HIF-1α 단백질은 집단간 차이를 보이지 않았다(p>.05). VEGF 단백질의 경우, 통제집단보다 정신 분열 집단에서 유의하게 낮게 나타났으며(p<.01), 정신분열+운동 집단의 VEGF가 정신분열 집단보다 유의하게 높게 나타났다(p<.01). 따라서 골격근 내 혈관생성 관련 단백질들은 정신분열 유도에 의해 낮아질 수 있으며 반대로 운동은 이를 개선하는 효과를 가지는 것으로 사료된다.

Purpose: This study examined the effects of α-lipoic acid in diluted solvents on cell growth in 3T3-L1 cells according to the treated concentration and times. Methods: Adipocyte 3T3-L1 cell were cultured. Confluent cells underwent starvation with SFM for 1 day and then were cultured in a medium containing various concentrations 0, 100, 200, and 400 μmol/L of α-lipoic acid. The cell viability was measured using the EZ Cytox assay kit. In addition, the effect of α-lipoic acid of diluted solvents on the cell growth in 3T3-L1cells was examined according to the treated concentration and times. Results: The α- lipoic acid diluted ethanol inhibited cell proliferation in a dose and time dependent manner. The α-lipoic acid diluted ethanol induced adipocyte 3T3-L1 cells proliferation with an adipocyte inducer. In addition, α-lipoic acid inhibited adipocyte 3T3- L1 growth in a dose and time dependent manner (p<0.05). Conclusion: This study showed that a treatment with α-lipoic acid diluted ethanol inhibits cell growth of, adipocyte 3T3-L1 cells induced with an adipocyte inducer, (200 μmol/L of α- lipoic acid) treated for 48 hr.

This study was performed to investigate the body fat-lowering effect of garlic powder in peroxisome proliferator-activated receptor γ coactivator-1 α (PGC-1α)-luciferase transgenic mice (TG). In this study, we generated transgenic mice with a PGC-1α promoter (—970/+412 bp) containing luciferase as a reporter gene. Mice were fed a 45% high-fat diet for 8 weeks to induce obesity. Subsequently, mice were maintained on either a high-fat control diet (CON), or high-fat diets supplemented with 2% (GP2) or 5% (GP5) garlic powder for an additional 8 weeks. Dietary garlic powder reduced the body weight in the GP2 and GP5 groups, compared to the CON group. Furthermore, garlic supplementation significantly decreased the plasma levels of triglycerides, total cholesterol, and leptin in the GP5 group, compared to the CON group. Specifically, luciferase activity in liver, white adipose tissue (WAT), and brown adipose tissue (BAT) was increased by garlic supplementation in a dose-dependent manner. These results suggest that the body fat-lowering effect of garlic powder might be related to PGC-1α by the increase in luciferase activity in liver, WAT, and BAT. Furthermore, transgenic mice might be useful for evaluating the body fat-lowering effect of various health functional foods.

Receptor activator of nuclear factor-κB ligand (RANKL) is an osteoblast/stromal cell-derived essential factor for osteoclastogenesis. During endochondral bone formation, hypertrophic chondrocytes calcify cartilage matrix that is subsequently resorbed by osteoclasts in order to be replaced by new bone. Hypoxia-induced upregulation of RANKL expression has been previously demonstrated in an in vitro system using osteoblasts; however, the involved mechanism remains unclear in chondrocytes. In the present study, we investigated whether hypoxia regulates RANKL expression in ATDC5 cells, a murine chondrogenic cell line, and hypoxiainducible factor-1α (HIF-1α) mediates hypoxia-induced RANKL expression by transactivating the RANKL promoter. The expression levels of RANKL mRNA and protein, as well as HIF-1α protein, were significantly increased in ATDC5 cells under hypoxic condition. Constitutively active HIF-1α alone significantly increased the levels of RANKL expression under normoxic conditions, whereas dominant negative HIF-1α reduced hypoxia-induced RANKL expression. HIF-1α increased RANKL promoter reporter activity in a HIF-1α binding element-dependent manner in ATDC5 cells. Hypoxia-induced RANKL levels were much higher in differentiated ATDC5 cells, as compared to proliferating ATDC5 cells. These results suggested that under hypoxic conditions, HIF-1α mediates induction of RANKL expression in chondrocytes; in addition, hypoxia plays a role in osteoclastogenesis during endochondral bone formation, at least in part, through the induction of RANKL expression in hypertrophic chondrocytes.

To overcome the hyperacute immune rejection during pig-to-non-human primates xenotranasplantation, we have produced and bred α-1,3-galactosyltransferase knock-out (GalT —/—) pigs. In this study, the somatic cells and tissues from the GalT —/— pigs were characterized by an analysis of the expression of Galα-1,3-Gal (α-Gal) epitope. Briefly, ear fibroblast cell lines of 19 homozygous GalT —/— pigs were established and cryopreserved. The expression of α-Gal epitope in the cells was measured by fluorescence activated cell sorter (FACS) analysis using BS-I-B4 lectin. Also, the homozygous (GalT —/—) cells and tissues samples were immunostained with BS-I-B4 lectin for analysis of α-Gal epitope expression. The results showed that the expression of α-Gal epitope in GalT —/— cells (0.2 %) were significantly (p< 0.05) down-regulated to the range of cynomolgus monkey fibroblast (0.2 %) cells compared to heterozygous (GalT —/+) (9.3 %) and wild type (GalT +/+) (93.7 %) fibroblast cells. In the immunostaining results, while the expression of α-Gal epitope was detected a partly in GalT —/+ cells and mostly in GalT +/+ cells, it was almost not detected in the GalT —/— cells. Also, immunostaining results from various tissues of the GalT —/— pig showed that the expression of α-Gal epitope was not detectable, whereas various tissues from GalT +/+ pig showed a strong expression of α-Gal epitope. Our results demonstrated that α-Gal epitope expressions from GalT —/— pigs were successfully knocked out to prevent hyperacute immune rejection for further study of xenotransplantation.

본 연구에서는 silicalite-1 제올라이트 분리막 합성 시에 종결정 코팅용액 pH 변화가 제올라이트 분리층 미세구조에 미치는 영항을 고찰하였다. 75 nm 크기로 합성된 종결정은 에탄올에 분산된 후 침지코팅법으로 지지체 표면에 코팅되었으며 분산용액의 pH는 2.2, 7.0, 9.3으로 조절되었다. pH가 7인 경우, 균일하고 두께가 3~4 μm인 silicalite-1 제올라이트 분리층이 형성되었고 분리층 결정입 크기는 100 nm로 미세하였다. 반면, pH가 2.2와 9.3인 경우, 분리층 두께가 얇고 불완전하였으며 분리층 결정입 크기도 약 1 μm로 조대하였다. pH 7에서 완전한 제올라이트 분리층이 형성된 것은 침지코팅 중에 지지체와 종 결정이 서로 다른 부호의 전하를 가져 정전기적 인력이 작용하여 균일하고 조밀하며 두껍고 다층의 종결정 코팅층이 형성되었 기 때문이었다. 반면에 pH가 2.2와 9.3인 경우, 침지코팅 중에 지지체와 종결정이 서로 같은 부호의 전하를 가져 정전기적 반 발력이 작용하기 때문에 불완전한 덮힘에 의하여 불완전한 분리층이 형성된다고 판단되었다. 결론적으로, 종결정 코팅용액의 pH가 silicalite-1 제올라이트 분리층의 두께, 결정립 크기 등 미세구조를 결정하는 중요한 인자임을 확인할 수 있었다.

Diabetes mellitus, the most common metabolic disorder, is divided into two types: type 1 and type 2. The essential treatment of type 1 diabetes, caused by immune-mediated destruction of β-cells, is transplantation of the pancreas; however, this treatment is limited by issues such as the lack of donors for islet transplantation and immune rejection. As an alternative approach, stem cell therapy has been used as a new tool. The present study revealed that bone marrowderived mesenchymal stromal cells (BM-MSCs) could be transdifferentiated into pancreatic cells by the insertion of a key gene for embryonic development of the pancreas, the pancreatic and duodenal homeobox factor 1 (PDX1). To avoid immune rejection associated with xenotransplantation and to develop a new cell-based treatment, BM-MSCs from α-1,3-galactosyltransferase knockout (GalT KO) pigs were used as the source of the cells. Transfection of the EGFP-hPDX1 gene into GalT KO pig-derived BM-MSCs was performed by electroporation. Cells were evaluated for hPDX1 expression by immunofluorescence and RT-PCR. Transdifferentiation into pancreatic cells was confirmed by morphological transformation, immunofluorescence, and endogenous pPDX1 gene expression. At 3∼4 weeks after transduction, cell morphology changed from spindle-like shape to round shape, similar to that observed in cuboidal epithelium expressing EGFP. Results of RT-PCR confirmed the expression of both exogenous hPDX1 and endogenous pPDX1. Therefore, GalT KO pig-derived BM-MSCs transdifferentiated into pancreatic cells by transfection of hPDX1. The present results are indicative of the therapeutic potential of PDX1-expressing GalT KO pig-derived BM-MSCs in β-cell replacement. This potential needs to be explored further by using in vivo studies to confirm these findings.

동물의 장기를 인간에게 이식하게 되면 초급성거부반응(Hyperacute rejection, HAR)이 일어난다. 초급성거부반응은 면역계의 구성요소 중 보체(complement)에 의해 일어나는 거부반응으로 돼지의 혈관세포 표면에 있는 Galα(1,3)Gal 당분자에 인간의 항체가 즉각 반응하기 때문에 일어나며, α1,3-galactosyltransferase(α1,3-GT) 유전자는 돼지 혈관세포 표면의 Galα(1,3)Gal 당분자 생성에 관여한다. 따라서 인간에게 돼지의 장기를 이식하기 위해서는 α1,3-galactosyltransferase 유전자를 제거하는 것이 필요한 것으로 알려져 있다. 본 연구실의 이전 연구에서, 시카고 미니돼지 귀체세포에서 상동 재조합(Homologous recombination)을 통해 α1,3-galactosyltransferase 유전자가 제거된 체세포를 개발한 바 있으며, 이 체세포를 통하여 α1,3-GT 유전자가 제거된 돼지도 생산된 바 있다. 본 연구에서는, human serum 처리 시 돼지 세포를 보호해 준다고 보고되고 있는 human complement regulator인 human Decay-accelerating factor(hDAF)와 human α1,2-fucosyltransferase(hHT)유전자를 α1,3-GT 유전자 위치에 gene targeting하여 동시에 hDAF와 hHT가 발현하는 체세포를 개발하였다. Knock-in vector는 hDAF와 hHT 두 유전자가 발현할 수 있도록 IRES로 연결하였으며, α1,3-GT 유전자의 start codon을 이용하여 발현할 수 있도록 구축하였다. 구축한 vector는 electroporation을 통해 미니돼지 체세포에 도입하였으며, PCR 결과, α1,3-GT 유전자 위치에서 상동 재조합이 일어났음을 확인하였다. Positivenegative 선별 방법을 통해 얻은 gene targeting 된 체세포는 RT-PCR에 의해 hDAF와 hHT 유전자의 발현이 확인되었으며, 대조군(NIH minipig)에 비해 α1,3-GT 유전자의 발현이 감소하였다. 또한 이들 세포에 100% human complement serum을 처리하였을 때 knock-in 세포가 대조군에 비해 30% 정도 더 높은 생존율을 보였다. 따라서 개발된 체세포는 이종간 장기이식을 위한 돼지 생산과 함께 이를 이용한 이종간의 장기 이식 시 초급성 거부반응을 억제하는 데 사용될 수 있을 것으로 생각된다.