• Journal of Periodontal and Implant Science
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• 제52권3호
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• pp.206-219
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• 2022

Purpose: This study was performed to evaluate the influence of local application of thymoquinone (TQ) on bone healing in experimental bone defects infected with Porphyromonas gingivalis (PG). Methods: Forty-two female rats were randomly divided into 6 groups. A bone defect was created on the right tibia of all animals. The PG, PG/collagen membrane (COL) and PG/TQ/COL groups were infected with PG. In the COL and PG/COL groups, the defects were covered with a COL; in the TQ/COL and PG/TQ/COL groups, the defects were covered with a TQ-containing COL. After 28 days, all animals were sacrificed. Quantitative measurements of new bone formation and osteoblast lining, as well as semiquantitative measurements of capillary density and tissue response, were analyzed. Furthermore, the presence of bacterial infections in defect areas was evaluated. Results: The new bone formation, osteoblast number, and capillary density were significantly higher in the TQ groups than in the control groups (P<0.001, P<0.001, and P<0.01, respectively). In a comparison between the TQ/COL group, with a TQ-containing COL (TQ/COL), and the PG-infected TQ-containing COL (PG/TQ/COL) group, the newly formed bone and capillary density were higher in the TQ/COL group (P<0.01). When the control group was compared to the PG, PG/COL, and PG/TQ/COL groups in terms of tissue response, the differences were statistically significant (P<0.001, P=0.02, and P=0.041, respectively). The intensity of the inflammatory cell reaction was higher in the PG, PG/COL, and PG/TQ/COL groups (P<0.05). Conclusions: Within the limitations of this study, the local application of a TQ-containing COL positively affected bone healing even if the bone defects were infected. The results suggest that TQ increased angiogenesis and showed promise for accelerating bone defect healing. Further research is warranted to support these findings and reach more definitive conclusions.

• Molecules and Cells
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• 제45권7호
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• pp.465-478
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• 2022

MicroRNAs (miRNAs) are a class of small non-coding RNAs that regulate the expression of target messenger RNA (mRNA) complementary to the 3' untranslated region (UTR) at the post-transcriptional level. Hsa-miR-422a, which is commonly known as miRNA derived from transposable element (MDTE), was derived from short interspersed nuclear element (SINE). Through expression analysis, hsa-miR-422a was found to be highly expressed in both the small intestine and liver of crab-eating monkey. AT-Rich Interaction Domain 5 B (ARID5B) was selected as the target gene of hsa-miR-422a, which has two binding sites in both the exon and 3'UTR of ARID5B. To identify the interaction between hsa-miR-422a and ARID5B, a dual luciferase assay was conducted in HepG2 cell line. The luciferase activity of cells treated with the hsa-miR-422a mimic was upregulated and inversely downregulated when both the hsa-miR-422a mimic and inhibitor were administered. Nuclear factor erythroid-2 (NF-E2) was selected as the core transcription factor (TF) via feed forward loop analysis. The luciferase expression was downregulated when both the hsa-miR-422a mimic and siRNA of NF-E2 were treated, compared to the treatment of the hsa-miR-422a mimic alone. The present study suggests that hsa-miR-422a derived from SINE could bind to the exon region as well as the 3'UTR of ARID5B. Additionally, hsa-miR-422a was found to share binding sites in ARID5B with several TFs, including NF-E2. The hsa-miR-422a might thus interact with TF to regulate the expression of ARID5B, as demonstrated experimentally. Altogether, hsa-miR-422a acts as a super enhancer miRNA of ARID5B by collaborating with TF and NF-E2.

• Korean Chemical Engineering Research
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• 제60권4호
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• pp.574-581
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• 2022

리튬-황 전지의 기능성 중간층으로 그래핀과 Mo₂C/Mo₂N 나노입자로 구성된 나노섬유(Mo₂C/Mo₂N rGO NFs)를 사용하였다. Mo₂C/Mo₂N 나노입자는 섬유 구조 내 고르게 분산되어 리튬 폴리설파이드의 화학적 흡착을 위한 활성 사이트 역할을 함으로써 전해질로의 용출을 효과적으로 억제하였다. 또한 구조 내 매트릭스로 구성된 그래핀 나노시트는 충방전이 진행되는 동안 이온 및 전자의 빠른 이동을 보장할 뿐만 아니라 반응 시 산화/환원 반응을 원활하게 하여 높은 리튬 폴리설파이드의 재사용을 보장하였다. 그 결과 Mo₂C/Mo₂N rGO NFs로 코팅된 분리막을 기능성 중간층으로 사용, 순수 황 전극(황 함량 70 wt%, 황 로딩 2.1 mg cm^-2)으로 제작된 리튬-황 전지는 0.1 C에서 400회 충방전 후 476 mA h g^-1의 안정적인 방전 용량을 나타냈으며, 1.0 C의 높은 전류밀도에서도 574 mA h g^-1의 방전용량을 나타내었다. 본 연구에서 제안된 나노구조체 합성 전략은 고성능 리튬-황 전지 용 기능성 중간층 및 다양한 에너지 저장 소재분야로의 확장이 가능하다.

• Journal of Plant Biotechnology
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• 제49권3호
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• pp.187-192
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• 2022

식물 캘러스의 월간 또는 주간의 반복적인 계대배양은 노동 집약적이며 모 세포주로부터 somaclonal variation 발생 위험을 증가시킨다. 식물 캘러스를 보존하기위한 가장 효과적인 방법은 액체질소에 저장하는 초저온동결보존 방법이다. 하지만 초저온동결보존 방법은 동일한 방법으로 여러 식물의 캘러스에 적용할 수 없어 보존 방법 개발에 많은 어려움이 따른다. 또한 해동 후 냉동되어진 캘러스의 생존과 생존 후 재생 속도가 불확실 하다는 단점이 있다. 그러므로 활성 상태의 식물 캘러스의 계대배양 간격을 증가시키는 방법의 개발이 필요하다. 본 연구에서는 냉동과정 없는 활성상태의 다양한 종의 식물 캘러스를 계대배양 없이 4주에서 12주 동안 15℃에서 배양하였다. 25℃에서 12주간 배양한 8종류의 식물 캘러스들은 모두 2배 이하의 성장을 보였으나 15℃에서 12주간 배양 조건에서는 8종류의 식물 캘러스들은 최소 2배 이상의 성장을 하였다. 또한 15℃에서 배양 후 25℃에서 회복시킨 캘러스들 사이의 항산화 활성 역시 크게 변화하지 변하지 않았다. 이러한 결과는 배지조성이나 특별한 새로운 과정없이 15℃ 저온으로 계대배양 간격을 증가시킬 수 있음을 보여준다. 또한 여러 식물 종의 캘러스들 모두에서 긍정적인 결과는 여러 캘러스들에 동일한 방법으로 계대배양 간격을 증가시킴으로 노동력 감소는 물론 somaclonal variation을 상대적으로 줄여 줄 것으로 예상한다.

• Journal of Microbiology and Biotechnology
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• 제31권8호
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• pp.1163-1174
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• 2021

Casein-derived antioxidant peptides by using microbial proteases have gained increasing attention. Combination of two microbial proteases, Protin SD-NY10 and Protease A "Amano" 2SD, was employed to hydrolyze casein to obtain potential antioxidant peptides that were identified by LC-MS/MS, chemically synthesized and characterized in a oxidatively damaged HepG2 cell model. Four peptides, YQLD, FSDIPNPIGSEN, FSDIPNPIGSE, YFYP were found to possess high 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging ability. Evaluation with HepG2 cells showed that the 4 peptides at low concentrations (< 1.0 mg/ml) protected the cells against oxidative damage. The 4 peptides exhibited different levels of antioxidant activity by stimulating mRNA and protein expression of the antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px), as well as nuclear factor erythroid-2-related factor 2 (Nrf2), but decreasing the mRNA expression of Kelch-like ECH-associated protein 1 (Keap1). Furthermore, these peptides decreased production of reactive oxygen species (ROS) and malondialdehyde (MDA), but increased glutathione (GSH) production in HepG2 cells. Therefore, the 4 casein-derived peptides obtained by using microbial proteases exhibited different antioxidant activity by activating the Keap1-Nrf2 signaling pathway, and they could serve as potential antioxidant agents in functional foods or pharmaceutic preparation.

• Journal of Veterinary Science
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• 제21권2호
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• pp.26.1-26.14
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• 2020

Pancreatic ductal adenocarcinoma is a lethal cancer type that is associated with multiple gene mutations in somatic cells. Genetically engineered mouse is hardly applicable for developing a pancreatic cancer model, and the xenograft model poses a limitation in the reflection of early stage pancreatic cancer. Thus, in vivo somatic cell gene engineering with clustered regularly interspaced short palindromic repeats is drawing increasing attention for generating an animal model of pancreatic cancer. In this study, we selected Kras, Trp53, Ink4a, Smad4, and Brca2 as target genes, and applied Campylobacter jejuni Cas9 (CjCas9) and Streptococcus pyogens Cas9 (SpCas9) for developing pancreatic cancer using adeno associated virus (AAV) transduction. After confirming multifocal and diffuse transduction of AAV2, we generated SpCas9 overexpression mice, which exhibited high double-strand DNA breakage (DSB) in target genes and pancreatic intraepithelial neoplasia (PanIN) lesions with two AAV transductions; however, wild-type (WT) mice with three AAV transductions did not develop PanIN. Furthermore, small-sized Cjcas9 was applied to WT mice with two AAV system, which, in addition, developed high extensive DSB and PanIN lesions. Histological changes and expression of cancer markers such as Ki67, cytokeratin, Mucin5a, alpha smooth muscle actin in duct and islet cells were observed. In addition, the study revealed several findings such as 1) multiple DSB potential of AAV-CjCas9, 2) peri-ductal lymphocyte infiltration, 3) multi-focal cancer marker expression, and 4) requirement of > 12 months for initiation of PanIN in AAV mediated targeting. In this study, we present a useful tool for in vivo cancer modeling that would be applicable for other disease models as well.

• Journal of Veterinary Science
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• 제23권6호
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• pp.88.01-88.14
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• 2022

Background: The olfactory mucosa (OM) is crucial for odorant perception in the main olfactory system. The terminal carbohydrates of glycoconjugates influence chemoreception in the olfactory epithelium (OE). Objectives: The histological characteristics and glycoconjugate composition of the OM of Korean native cattle (Hanwoo, Bos taurus coreae) were examined to characterize their morphology and possible functions during postnatal development. Methods: The OM of neonate and adult Korean native cattle was evaluated using histological, immunohistochemical, and lectin histochemical methods. Results: Histologically, the OM in both neonates and adults consists of the olfactory epithelium and the lamina propria. Additionally, using periodic acid Schiff and Alcian blue (pH 2.5), the mucus specificity of the Bowman's gland duct and acini in the lamina propria was determined. Immunohistochemistry demonstrated that mature and immature olfactory sensory neurons of OEs express the olfactory marker protein and growth associated protein-43, respectively. Lectin histochemistry indicated that numerous glycoconjugates, including as N-acetylglucosamine, mannose, galactose, N-acetylgalactosamine, complex type N-glycan, and fucose groups, were expressed at varied levels in the different cell types in the OMs of neonates and adults at varying levels. According to our observations, the cattle possessed a well-developed olfactory system, and the expression patterns of glycoconjugates in neonatal and adult OMs varied considerably. Conclusions: This is the first study to describe the morphological assessment of the OM of Korean native cattle with a focus on lectin histochemistry. The findings suggest that glycoconjugates may play a role in olfactory chemoreception, and that their labeling properties may be closely related to OM development and maturity.

• 대한본초학회지
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• 제28권1호
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• pp.43-50
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• 2013

Objectives : Hwanggeumjakyak-tang (huangqin shaoyao tang, HJT) has been used to treat acute enteritis in traditional oriental medicine. However, there has been a lack of studies regarding the effects of HJT on the inflammatory activities and effector inflammatory disease mechanism about macrophage before is not known. So we examined the effect of HJT water extract on pro-inflammatory mediators in lipopolysaccharide (LPS) - stimulated mouse macrophage, RAW 264.7 cells. Methods : Cells were treated with 2 ug/mL of LPS 1 h prior to the addition of HJT. Cell viability was measured by MTS assay. The production of nitric oxide (NO) was determined by reacting cultured medium with Griess reagent. The expression of cyclooxygenase-2 (COX-2), inducible NO synthase (iNOS) and mitogen-activated protein kinases (MAPKs) was investigated by Western blot, RT-PCR. The content of level of cytokines (prostaglandin (PG) $E_2$, interleukin (IL)-6, IL-12, Tumor necrosis factor-alpha (TNF-${\alpha}$) and monocyte chemoattractant protein-1 (MCP-1)) in media from LPS-stimulated Raw 264.7 cells was analyed by ELISA kit. Results : HJT inhibited the production of NO, $PGE_2$, IL-6 as well as the expressions of iNOS, COX-2 but did not inhibit the production of IL-12, TNF-${\alpha}$, MCP-1 in the murine macrophage, RAW 264.7 cells. HJT also had suppression effects of LPS-induced MAPKs activation Conclusion : These results suggest that HJT has an anti-inflammatory therapeutic potential, which may result from inhibition of MAPK phosphorylation, thereby decreasing the expression of pro-inflammatory genes.

• 대한본초학회지
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• 제28권4호
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• pp.7-16
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• 2013

Objectives : Brassica campestris var narinosa (BCN), Canavalia gladiata DC semen (CGD) and their combinational prescription (BCN+CGD) have been use to demonstrate to regulate hematopoiesis. In the current study, we investigated whether Brassica campestris var narinosa, Canavalia gladiata DC semen and their combinational prescription is related to hemato-potentiating function using Sca-$1^+$ hematopoietic stem cells (Sca-$1^+HSCs$) as a testing system. Methods : Sca-$1^+HSCs$ isolated from femur in C57bl/6 mice with leukopenia and thrombocytopenia induced by cyclophosphamide (CTX). Then, Real-time PCR was performed to measure the mRNA expression, ELISA and haematopoiesis-related gene (EPO, TPO, IL-3, SCF, c-kit, GM-CSF), the phosphorylation of JAK2, GATA-1 and STAT-5a/b were observed by western blot, and the numbers of $CD117^+/Sca-1^+$ cell and the number of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E), semisolid clonogenic assay was performed. Result : When Sca-$1^+HSCs$ were treated with Brassica campestris var narinosa, Canavalia gladiata DC semen and their combinational prescription with rIL-3/rSCF, the expression of haematopoiesis-related (EPO, TPO, IL-3, SCF, c-kit, and GM-CSF) were significantly increased at the levels of mRNA as well as production in Sca-$1^+HSCs$. Additionally, CGS enhanced phosphorylation of JAK2, GATA-1, and signal transducer and activator of transcription-5a/b (STAT-5a/b) in Sca-$1^+HSCs$. Furthermore, their combinational prescription (BCN+CGD) significantly enhanced the growth rate of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E) in vitro. Conclusion : These result suggest that Brassica campestris var narinosa (BCN) and Canavalia gladiata DC have hematopoietic enhancement via hematopoietic cytokine-mediated JAK2/GATA-1/STAT-5a/b pathway, and their combinational prescription (BCN+CGD) has superior hematopoietic enhancement to those of individual extracts.

• 대한본초학회지
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• 제32권2호
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• pp.77-85
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• 2017

Objectives : This study aimed to investigate the unknown mechanisms behind the anti- inflammatory activity of Hyeonto-dan(HT) 70% ethanol extract on LPS-stimulated RAW 264.7 cells. Methods : Cells were treated with Hyeonto-dan 1 h prior to addition of 200 ng/mL of LPS. Cell viability was measured by the MTS assay. Nitric oxide levels were determined by the Griess assay. $PGE_2$ were measured using EIA kit. Pro-inflammatory cytokine production was measured by the enzyme-linked immunosorbent assay (ELISA). The expression of COX-2, iNOS, and MAPKs was investigated by Western blot, qRT-PCR. $NF-{\kappa}B$/p65 localization and interaction of the TLR-4 receptor with LPS was examined by immunofluorescence assays. Results : Hyeonto-dan had no cytotoxicity at the measured concentration. Hyeonto-dan inhibited NO production and pro-inflammatory cytokines such as IL-6, $TNF-{\alpha}$, and PGE2 as well as the protein and mRNA expression of iNOS and COX-2. Moreover, Hyeonto-dan inhibited the interaction between LPS and TLR-4 in murine macrophages. It suppressed phosphorylation of extracellular signal-regulated kinase (ERK 1/2), c-jun N-terminal kinase (JNK 1/2) and p38. Finally, it inhibited translocation of $NF-{\kappa}B$ in response to competitive LPS. Conclusions : Based on the results of this study, Hyeonto-dan inhibited the binding of TLR-4 receptor to LPS and inhibited the phosphorylation of extracellular signaling pathway MAPKs. These inhibitory effects are thought that the amount of $NF-{\kappa}B$ delivered to the nucleus was decreased and the inflammatory reaction was prevented by decreasing the production of LPS-induced $PGE_2$, NO, IL-6 and $TNF-{\alpha}$.