• 한방재활의학과학회지
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• 제32권3호
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• pp.1-11
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• 2022

Objectives This study was designed to evaluate the healing effect of Cordyceps Militaris (CM) on collagen II-induced arthritis rats. Methods Sprague-Dawley rats were randomly divided into 6 groups (normal, control, positive control, CM with low/medium/high dosage each). Type II collagen mixed with complete Freund's adjuvant (with 1:1 v/v) was injected subcutaneously, and the mixture was injected in a same manner one week after the first injection to boost arthritis. Arthritis index, paw thickness and von Frey test were conducted to observe physical changes. hematoxylin and eosin (H&E) staining was performed to observe knee cartilage. The levels of messenger RNA (mRNA) expressions of interleukin (IL)-1𝛽, IL-6, tumor necrosis factor-alpha (TNF-𝛼) in spleen were assessed by real-time polymerase chain reaction. Results Rheumatoid arthritis is an autoimmune disease that occurs on multiple joints and can lead to temporary shape change of bones or organ failure in severe cases. Here, we aimed to determine the effect of CM extract on rheumatoid arthritis by measuring paw thickness, arthritis index, conducting von Frey test and H&E staining, and evaluating the level of IL-1𝛽, IL-6, TNF-𝛼. As a result, paw thickness, arthritis index significantly decreased in low concentration group, hind leg became less sensitive in all expermental groups. Also, histological analysis showed that the damage of knee cartilage was prevented in all experimental groups. The level of mRNA of IL-1𝛽, IL-6, and TNF-𝛼 in spleen was analyzed to decide the effectiveness of CM extract. IL-1𝛽 did not show significant change, but IL-6 and TNF-𝛼 showed significant decrease in at least one of the experimental groups. Conclusions CM showed protective effect on knee tissue destruction and improved the physical conditions of the leg involving arthritis. Also, it showed that CM has anti-inflammatory effect on specific cytokines inducing rheumatoid arthritis. In conclusion, this study demonstrated that the therapeutic potential of CM for the treatment rheumatoid arthritis, and set the foundation for the further studies.

• Journal of Veterinary Science
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• 제21권5호
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• pp.80.1-80.13
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• 2020

Background: In suckling piglets, transmissible gastroenteritis virus (TGEV) causes lethal diarrhea accompanied by high infection and mortality rates, leading to considerable economic losses. This study explored methods of preventing or inhibiting their production. Bovine antimicrobial peptide-13 (APB-13) has antibacterial, antiviral, and immune functions. Objectives: This study analyzed the efficacy of APB-13 against TGEV through in vivo and in vitro experiments. Methods: The effects of APB-13 toxicity and virus inhibition rate on swine testicular (ST) cells were detected using 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT). The impact of APB-13 on virus replication was examined through the 50% tissue culture infective dose (TCID₅₀). The mRNA and protein levels were investigated by real-time quantitative polymerase chain reaction and western blot (WB). Tissue sections were used to detect intestinal morphological development. Results: The safe and effective concentration range of APB-13 on ST cells ranged from 0 to 62.5 ㎍/mL, and the highest viral inhibitory rate of APB-13 was 74.1%. The log₁₀TCID₅₀ of 62.5 ㎍/mL APB-13 was 3.63 lower than that of the virus control. The mRNA and protein expression at 62.5 ㎍/mL APB-13 was significantly lower than that of the virus control at 24 hpi. Piglets in the APB-13 group showed significantly lower viral shedding than that in the virus control group, and the pathological tissue sections of the jejunum morphology revealed significant differences between the groups. Conclusions: APB-13 exhibited good antiviral effects on TGEV in vivo and in vitro.

• Journal of Korean Neurosurgical Society
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• 제48권1호
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• pp.1-7
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• 2010

Objective : Notochordal cells in the intervertebral disc interact with nucleus pulposus (NP) cells and support the maintenance of disc homeostasis by regulation of matrix production. However, the influence of notochordal cells has not been evaluated in the annulus fibrosus (AF), which is the primary pain generator in the disc. We hypothesized that the notochordal cell has the capacity to modulate inflammatory mediators secreted by AF cells secondary to stimulation. Methods : Notochordal and AF cells were isolated from adult New Zealand white rabbits. AF pellets were cultured with notochordal cell clusters or in notochordal cell-conditioned media (NCCM) for 24 or 48 hours with proinflammatory cytokines at varying concentrations. Gene expression in AF pellets were assayed for nitric oxide synthase (iNOS), cyclo-oxygenase (COX)-2, and interleukin (IL)-6 by real time reverse transcriptase polymerase chain reaction (RT-PCR). Results : AF pellet in NCCM significantly decreased the iNOS and COX-2 messenger ribonucleic acid (mRNA) levels compared to AF pellets alone and AF pellets with notochordal cells (p < 0.05). AF pellet resulted in dose-dependent iNOS and COX-2 expression in response to IL-$1{\beta}$, stimulation, demonstrating that 1 ng/ml for 24 hours yielded a maximal response. AF pellet in NCCM significantly decreased the expression of iNOS and COX-2 in response to 1ng/ml IL-$1{\beta}$, stimulation at 24 hours (p < 0.05). There was no difference in IL-6 expression compared to AF pellets alone or AF pellets with notochordal cell clusters. Conclusion : We conclude that soluble factors from notochordal cells mitigate the gene expression of inflammatory mediators in stimulated AF, as expected after annular injury, suggesting that notochordal cells could serve as a novel therapeutic approach in symptomatic disc development.

• Genomics & Informatics
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• 제11권3호
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• pp.142-148
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• 2013

SINE-VNTR-Alu (SVA) elements are present in hominoid primates and are divided into 6 subfamilies (SVA-A to SVA-F) and active in the human population. Using a bioinformatic tool, 22 SVA element-associated genes are identified in the human genome. In an analysis of genomic structure, SVA elements are detected in the 5′ untranslated region (UTR) of HGSNAT (SVA-B), MRGPRX3 (SVA-D), HYAL1 (SVA-F), TCHH (SVA-F), and ATXN2L (SVA-F) genes, while some elements are observed in the 3′UTR of SPICE1 (SVA-B), TDRKH (SVA-C), GOSR1 (SVA-D), BBS5 (SVA-D), NEK5 (SVA-D), ABHD2 (SVA-F), C1QTNF7 (SVA-F), ORC6L (SVA-F), TMEM69 (SVA-F), and CCDC137 (SVA-F) genes. They could contribute to exon extension or supplying poly A signals. LEPR (SVA-C), ALOX5 (SVA-D), PDS5B (SVA-D), and ABCA10 (SVA-F) genes also showed alternative transcripts by SVA exonization events. Dominant expression of HYAL1_SVA appeared in lung tissues, while HYAL1_noSVA showed ubiquitous expression in various human tissues. Expression of both transcripts (TDRKH_SVA and TDRKH_noSVA) of the TDRKH gene appeared to be ubiquitous. Taken together, these data suggest that SVA elements cause transcript isoforms that contribute to modulation of gene regulation in various human tissues.

• International Journal of Oral Biology
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• 제41권1호
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• pp.25-32
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• 2016

The tongue has 4 kinds of papillae, which are filiform, fungiform (FU), foliate (FO) and circumvallate papilla (CV). Tongue papillae except filiform papilla include taste buds. The papillae differ in taste sensitivities, likely due to differential expression of taste receptors. In this study, we evaluated differences in the expression levels of taste receptors in FU, FO and CV. Male DBA2 mice, 42-60 days old, were used in the study. Messenger RNAs were extracted from the murine epithelial tissues including FU, FO and CV. Cloned DNAs were synthesized by reverse transcription. Quantitative PCRs (qPCRs) were performed to determine mRNA expression levels of taste receptors. Results of qPCR revealed that the relative expression levels and patterns were different among FU, FO and CV. All three type 1 taste receptors were expressed FU, FO and CV at varying relative expression levels. All 35 kinds of type 2 taste receptors showed higher expression in FO and CV than in FU. Tas2r108 and Tas2r137 showed the two highest expression levels in all tested papillae. The differential expression levels and patterns of taste receptors among the three papillae could contribute to the different physiological sensitivities by tongue areas. Additional studies such as in situ hybridization or taste receptor cell activity recording is necessary to elucidate the functional relationship between expression levels of taste receptors and taste sensitivity.

• 한국발생생물학회지:발생과생식
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• 제11권3호
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• pp.253-261
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• 2007

Lactadherin은 모유의 유지방구막의 당단백질의 하나로 mucin, butyrophilin과 결합된 복합체이다. 특히 모유중의 mucin과 lactadherin은 출생 직후 면역력이 약한 유아를 병균의 침입으로부터 효율적으로 방어하여 초기 유아의 생존과 성장 및 발달에 매우 중요한 역할을 수행한다. Lactadherin은 유아 설사의 원인이 되는 rotavirus의 번식과 성장을 억제한다. 아울러 이 단백질은 새로운 혈관의 형성을 촉진하는 주요한 단백질로 알려져 있으며, 이 단백질의 결핍이 치매의 발생과 관련되는 것으로 보고되고 있다. 본 연구는 이처럼 중요성이 강조되는 lactadherin에 대한 생화학적 및 생리학적인 연구를 하기 위한 기초연구를 진행하였다. 한국 여성의 유선조직에서 mRNA를 분리하였고, 1.2 kb lactadherin cDNA 유전자를 cloning하여 염기서열과 아미노산 배열을 결정하였다. 이 cDNA를 pET vector에 삽입하여 E. coli에서 43 kD 단백질을 발현시켰으며 Western blot으로 확인하였으며, 이 단백질을 정제하여 토끼에서 항체를 생산하여, 한국 여성의 모유에서 발현되는 70, 55, 46, 30 kD의 band를 확인하였다. 아울러 백인 여성의 lactadherin 유전자와 한국 여성의 정상 및 유방암 조직의 유전자 비교에서 다양한 SNP가 관찰되었고 변이의 다형성이 관찰되었다.

• The Korean Journal of Physiology and Pharmacology
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• 제23권2호
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• pp.141-150
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• 2019

Despite increased evidence of bio-activity following far-infrared (FIR) radiation, susceptibility of cell signaling to FIR radiation-induced homeostasis is poorly understood. To observe the effects of FIR radiation, FIR-radiated materials-coated fabric was put on experimental rats or applied to L6 cells, and microarray analysis, quantitative real-time polymerase chain reaction, and wound healing assays were performed. Microarray analysis revealed that messenger RNA expressions of rat muscle were stimulated by FIR radiation in a dose-dependent manner in amount of 10% and 30% materials-coated. In 30% group, 1,473 differentially expressed genes were identified (fold change [FC] > 1.5), and 218 genes were significantly regulated (FC > 1.5 and p < 0.05). Microarray analysis showed that extracellular matrix (ECM)-receptor interaction, focal adhesion, and cell migration-related pathways were significantly stimulated in rat muscle. ECM and platelet-derived growth factor (PDGF)-mediated cell migration-related genes were increased. And, results showed that the relative gene expression of actin beta was increased. FIR radiation also stimulated actin subunit and actin-related genes. We observed that wound healing was certainly promoted by FIR radiation over 48 h in L6 cells. Therefore, we suggest that FIR radiation can penetrate the body and stimulate PDGF-mediated cell migration through ECM-integrin signaling in rats.

• 대한치과교정학회지
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• 제49권5호
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• pp.299-309
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• 2019

Objective: This study aimed to investigate the effect of pre-applied orthodontic force on the regeneration of periodontal ligament (PDL) tissues and the underlying mechanisms in tooth replantation. Methods: Orthodontic force (50 cN) was applied to the left maxillary first molars of 7-week-old male Sprague-Dawley rats (n = 32); the right maxillary first molars were left untreated to serve as the control group. After 7 days, the first molars on both sides were fully luxated and were immediately replanted in their original sockets. To verify the effects of the pre-applied orthodontic force, we assessed gene expression by using microarray analysis and real-time reverse transcription polymerase chain reaction (RT-PCR), cell proliferation by using proliferating cell nuclear antigen (PCNA) immunofluorescence staining, and morphological changes by using histological analysis. Results: Application of orthodontic force for 7 days led to the proliferation of PDL tissues, as verified on microarray analysis and PCNA staining. Histological analysis after replantation revealed less root resorption, a better arrangement of PDL fibers, and earlier regeneration of periodontal tissues in the experimental group than in the control group. For the key genes involved in periodontal tissue remodeling, including CXCL2, CCL4, CCL7, MMP3, PCNA, OPG, and RUNX2, quantitative RT-PCR confirmed that messenger RNA levels were higher at 1 or 2 weeks in the experimental group. Conclusions: These results suggest that the application of orthodontic force prior to tooth replantation enhanced the proliferation and activities of PDL cells and may lead to higher success rates with fewer complications.

• 한방재활의학과학회지
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• 제31권4호
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• pp.1-11
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• 2021

Objectives Hwanggeumjakyak-tang (HJT) has traditionally been used to treat gastrointestinal inflammatory diseases; however, its protective effects against neuronal inflammation are still undiscovered. Methods We investigated the anti-neuroinflammatory effects of HJT water extract on lipopolysaccharide (LPS)-stimulated BV2 mouse microglia cells. BV2 cells were treated with LPS (1 ㎍/mL) 1 hour prior to the addition of HJT. We measured cell viability using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and nitrite production using the Griess assay. We performed a reverse transcription-polymerase chain reaction assay to measure messenger RNA expression of inflammatory cytokines including interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α. Western blot analysis was performed to determine protein expression of mitogen-activated protein kinases (MAPKs) and inhibitor of nuclear factor kappa B (NF-κB)α. Results HJT inhibited excessive nitrite release in LPS-stimulated BV2 cells and also significantly inhibited inflammatory cytokines such as IL-1β, IL-6, and TNF-α in LPS-stimulated BV2 cells. Moreover, HJT significantly suppressed LPS-induced MAPK and NF-κB activation and inhibited the elevation of IL-1β, IL-6, and TNF-α in the brain of LPS-injected mice. Conclusions Our study highlights the anti-neuroinflammatory effects of HJT via MAPK and NF-κB deactivation.

• Journal of Nutrition and Health
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• 제54권4호
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• pp.335-341
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• 2021

Purpose: Muscle mitochondria play a key role in regulating fatty acid and glucose metabolism. Dysfunction of muscle mitochondria is associated with metabolic diseases such as obesity and type 2 diabetes. Isorhamnetin (ISOR), also known as 3-O-methylquercetin, a quercetin metabolite, is a naturally occurring flavonoid in many plants. This study evaluated the effects of ISOR on the regulation of the mitochondrial function of C2C12 muscle cells. Methods: C2C12 muscle cells were differentiated for 5 days, and then treated in various concentrations of ISOR. Cytotoxicity was determined by assessing cell viability using the water-soluble tetrazolium salt-8 assay principle at different concentrations of ISOR and time points. Levels of the mitochondrial DNA (mtDNA) content and gene expression were measured by quantitative real-time polymerase chain reaction. The citrate synthase (CS) activity was quantified by the enzymatic method. Results: ISOR at a concentration of 10 µM did not show any cytotoxic effects. ISOR increased the mtDNA copy number in a time- or dose-dependent manner. The messenger RNA levels of genes involved in mitochondrial function, such as peroxisome proliferator-activated receptor-γ coactivator-1α, and uncoupling protein 3 were significantly stimulated by the ISOR treatment. The CS activity was also significantly increased in a time- or dose-dependent manner. Conclusion: These results suggest that ISOR enhances the regulation of mitochondrial function, which was at least partially mediated via the stimulation of the mtDNA replication, mitochondrial gene expression, and CS activity in C2C12 muscle cells. Therefore, ISOR may be useful as a potential food ingredient to prevent metabolic diseases-associated muscle mitochondrial dysfunction.