• 대한한의학회지
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• 제30권3호
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• pp.51-60
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• 2009

Objectives : It has been known that immune reaction plays an important role in the pathogenesis of otitis media (OM). We investigated the change of middle ear mucosal inflammation induced by lipopolysaccharide (LPS) following administration of Taglisodog-eum (TSE) in experimental animals. Materials and Methods : Otitis media was induced by injecting 1 mg/kg of Pseudomonas aeruginosa LPS transtympanically in 60 rats. These rats were divided into two groups, the LPS group (n=30, remained naive after OM elicitation) and the TSE group (n=30, treated with TSE after OM elicitation) and 6 additional rats were kept as a normal control group (n=6, remained naive until tissue collection). The rats were killed at the 1st, 3rd, and 7th days after challenge. The temporal bones in each group were harvested and examined histopathologically by hematoxyline-eosin stain. Middle ear mucosa were taken at the 1st, 3rd, and 7th days after challenge. The levels of spicing variants of TNF-${\alpha}$ transcription were evaluated by Realtime-PCR. Results : TSE suppressed LPS-induced TNF-${\alpha}$ mRNA expression and thickness of the submucosal layer and infiltration of inflammatory cells in rat middle ear epithelium. Conclusion : The results suggest that TSE may be effective in decreasing inflammation with particular application to mucosal metaplasia in OM.

• 한방재활의학과학회지
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• 제28권2호
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• pp.37-45
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• 2018

Objectives This study was designed to investigate whether the Gagamtongsoon-San (GT) has an inhibitory effect and its mechanisms are associated with the iNOS and COX-2. Methods Cytotoxic activity of GT extract on RAW264.7 cells was evaluated by using 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) solution. Inflammatory condition was induced by LPS. NO production was measured using Griess reagent system. The expressions of iNOS and COX-2 mRNA and protein were determined by realtime PCR. The concentrations of PGE2 were measured by an enzyme immunoassay (EIA). Results The GT does not impair the cell viability in tested concentration $500{\mu}g/ml$ or below. GT significantly reduced the NO production in a dose-dependent manner. GT $500{\mu}g/ml$ also suppressed LPS-induced mRNA expressions of iNOS and COX-2. GT $500{\mu}g/ml$ reduced the PGE2 secretion in LPS induced RAW264.7 cells. Conclusions These outcomes show that GT extract has an anti-inflammatory activities. And also this conclusion can be the data that supports the GT's anti-inflammatory effect objectively.

• 핵의학기술
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• 제23권1호
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• pp.35-39
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• 2019

[목적] B형 간염바이러스(hepatitis B virus, HBV)감염은 전세계적으로 중요한 공중 보건 문제이며 만성간염, 간 경변, 간암의 주요 원인으로 알려져 있으며, 이러한 질환의 진단 및 치료에 B형 간염바이러스의 혈청학적 검사는 필수적이다. 항 바이러스 치료중인 B형 간염 환자를 대상으로 면역방사계수 측정법(IRMA; Immunoradiometric assay)과 화학발광 미세입자 면역분석법(CMIA; Chemiluminescent Micropartical Immunoassay)을 이용하여 HBe-Ag 검사를 시행하였고, 실시간 중합효소 연쇄반응(RT-PCR; Realtime-Polymerase Chain Reaction)법을 이용하여 혈청 내 HBV DNA 검출율 을 비교 분석 하였다. [대상 및 방법] 항 바이러스 치료가 시행중인 B형 간염 환자 270명을 대상으로 HBeAg 혈청 검사와 HBV DNA 정량 검사를 실시하였다. HBeAg 혈청 검사는 검출 원리가 다른 두 가지 혈청학적 검사법(IRMA, CMIA)을 적용 하였고, 혈청 내 HBV DNA는 Abbott m2000 System을 사용하여 실시간 중합효소 연쇄반응(RT-PCR; Realtime-Polymerase Chain Reaction)법으로 정량 측정 하였다. [결과] HBeAg 검출율은 면역방사계수법(IRMA)의 경우 24.1% (65/205), 화학발광 미세입자 면역 분석법(CMIA)에서는 82.2% (222/48)의 결과를 보였다. 혈청학적 검사방법(IRMA, CMIA)에 따른 HBeAg 검사결과의 일치율은 33% (89/270)이다. 실시간 중합효소 연쇄반응(RT-PCR)을 이용한 혈청 내 HBV DNA의 검출율은 29.3% (79/191)를 보였고, 혈청 내 HBV-DNA 농도는 $16IU/mL{\times}1.0{\times}10^9IU/mL$ 이며 검출한계는 <15IU/mL 이다. 면역방사계수법(IRMA)으로 HBeAg 검출결과가 양성일때 55.4%, 그리고 음성일때 20.9%의 HBV DNA 검출율과 $1.1{\times}10^8IU/mL$, $5.7{\times}10^5IU/mL$의 혈청내 HBV DNA농도를 나타냈다. 이에 반해 화학발광 미세입자 면역 분석법(CMIA)의 경우 HBeAg 검출결과가 양성일때 HBV DNA 검출율은 28.4%, 음성일때 33.3%의 결과를 나타냈으며 혈청 내 HBV DNA농도는 $6.0{\times}10^7IU/mL$, $2.4{\times}10^5IU/mL$ 이었다. 면역방사계수법과 화학발광 미세입자 면역 분석법에서 동일하게 HBeAg 검출 결과가 양성인 경우 HBV DNA 검출율은 62.3%의 결과를 보였으며, 혈청 내 HBV DNA 농도는 $1.1{\times}10^8IU/mL$ 이다. [결론] 혈청학적 검사법에 따른 HeAg 검출율은 많은 차이를 보였다. 이러한 차이는 검사kit에 사용된 Ab의 특성과 epitope, HBV의 genotype등 여러 가지 원인으로 생각된다. 혈청학적 검사 결과로 분류 된 그룹별 HBV DNA의 검출율과 농도를 비교한 결과, Group II(IRMA 양성, CMIA 양성, N=53)에서 높은 검출율과 농도를 확인할 수 있었다.

• 동의생리병리학회지
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• 제17권3호
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• pp.633-642
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• 2003

A pain is the symptom which defends against noxious stimulus about a human body, it is known that if the periphery of perceptive nerve were stimulated by a physical or chemical factors, the stimulation is induced by transmission to pain center in the cerebral cortex according to pain conduction tract. The treatment of pain is to decrease a stimulus that causes a pain or block off a nerve transmitting a stimulus or puts on a way to calm down pain center, but It is for adjustment of a pain to be the most representative in acupuncture among various ways to cure a pain in Oriental medicine. However, the analgesic effect of an individual response to acupuncture stimulation shows marked individual variations, so these days genetic a few approach is attempted. On this the author determined that the responding group was appointed those whose tail flick latency (TFL) responding time delayed the minimum of 30 % comparing with basal reaction time. For those whose TFL time had shorter than 30 % was grouped as a non-responding group. And then the hypothalamus of each group was dissected and RNA was further purified. After synthesizing cDNA using oligo dT primer, products were finally applied to the PCR. The results were as follows; The ratio of responding group to non-responding group was 6:4. Ach T (acetylcholinesterase T subunit), BF-I (Brain factor-I), DBH (Dopamine β-hydroxylase) and PNM (Phosphotidylethanolamine N-Methyltransferase) were revealed significantly in the responding group. Cathepsin B and Tau were revealed significantly in the non-responding group. The PCR results show that Ach T, BF-I, DBH and PNM are expressed abundantly in the responding group, where as cathepsin B and tau are abundant in the non-responding group. These results suggest that the analgesic effect on acupuncture stimulation is related to regulation of neurotransmitter as well as neurodegeration of cerebrum.

• 한국임상수의학회지
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• 제26권5호
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• pp.433-440
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• 2009

High levels of inflammatory cytokines were proposed contributors to the pathogenesis of a various inflammatory skin disorders. Therefore, investigating the immune response of the inflammatory skin disorder allows a better understanding of pathogenesis of a various inflammatory skin disorders and therapeutic approaches. The aim of this study was to analyze of the immune response in dogs with chronic inflammatory skin disease. To this aim, the present study evaluated relative mRNA expression of canine $IFN-{\gamma}$, IL-4, $TGF-{\beta}$ and IL-10 using TaqMan realtime PCR assays and semi-quantitative RT-PCR in freshly isolated peripheral blood mononuclear cells from twenty dogs with chronic inflammatory skin disease and ten normal dogs. The relative mRNA expression levels of IL-4 mRNA were significantly higher in dogs with chronic inflammatory skin disease than those in normal dogs (P < 0.01). The results of present study also showed a tendency towards increased expression of IL-10 transcripts in dogs with chronic inflammatory skin disease. However, there were no significant differences in the levels $IFN-{\gamma},\;TGF-{\beta}$ between normal and chronically inflammed dogs. In addition, the concentration of serum IgE was significantly increased in dogs with chronic inflammatory skin disease compared with those in normal dogs (P < 0.01). In histopathological examination, we found that there were markedly increased mast cell counts in chronically inflammed dogs (P < 0.05). These results suggest that the pathogenesis of chronic inflammatory skin disease might be associated with a T-cell mediated inflammatory responses characterized by a Th2-skewed immune response. Based on these results, the modulation of Th1/Th2 balance may be an effective therapeutic strategy for the treatment of chronic inflammatory skin disease.

• 생명과학회지
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• 제29권6호
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• pp.631-636
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• 2019

본 연구에서는 산화질소 유도성(nitric oxide-induced, NOI) 유전자를 건조 처리된 고구마 실뿌리의 EST 라이브러리에서 분리하였다. 분리된 IbNOI 유전자의 cDNA는 712 bp의 길이이며, 77개의 아미노산으로 구성되어 있었다. Blast 데이터베이스를 분석한 결과, 식물에서 보고된 NOI 단백질에 속하는 것으로 확인되었다. RT-PCR과 realtime-PCR을 통해 IbNOI 유전자의 발현수준을 고구마 식물체의 조직별로 조사한 결과, 저장뿌리와 현탁배양세포에서 높은 발현 수준을 보였다. 잎에서 산화질소를 유도하는 SNP와 화합물 스트레스들을 처리시, IbNOI의 발현이 증가함을 확인할 수 있었다. 또한 고염, 건조와 같은 비생물학적 스트레스 및 병원균 감염에 의해서도 IbNOI의 발현이 유도됨을 확인할 수 있었다. 본 연구의 결과들을 통해 IbNOI 유전자는 다양한 비생물학적 스트레스 및 병원균 감염 동안 산화질소와 연관된 조절기작을 통해 식물의 방어기능에 관여할 것으로 생각되는 바이다.

• 생명과학회지
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• 제18권8호
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• pp.1090-1095
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• 2008

근내지방 축적이 왕성하게 진행되는 12개월 및 27개월령 사이에 유전자발현이 증가하는 inducible cAMP early repressor (ICER) 유전자의 클로닝 및 유전자발현 특성을 규명하기 위하여 한우조직별, 근육 내 조지방 함량에 따라 마블링 high- 및 low 한우 각 4두와 3두씩을 공시하였다. ddRT-PCR 분석을 통하여 증폭된 300 bp PCR 산물 및 EST 서열을 중심으로 1.5 kb 한우 ICER cDNA를 염기서열 분석하였으며, 조직별 유전자발현분석결과, 식이지방이 흡수되는 소장 조직에서 ICER 유전자의 발현량이 가장 높았다. 아울러, 같은 근육타입인 등심 및 우둔조직 사이에서 ICER 유전자발현은 등심조직에서 2.5배 많이 발현하였다. 한우 마블링 high, low 두 그룹 간 유전자발현 분석 결과, high 그룹에서 ICER 유전자발현이 4배 이상 증가하는 것으로 분석되었다. 따라서, ICER 유전자는 가축의 마블링과 밀접한 관련이 있는 유전자임이 사료된다.

• Clinical and Experimental Pediatrics
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• 제53권5호
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• pp.616-622
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• 2010

Enterovirus 71 (EV71) has been recognized as a frequent cause of epidemics of hand-foot-and-mouth disease (HFMD) associated with severe neurological symptoms. In the spring of 2009, HFMD was epidemic in Korea. Severe cases with complication, including death, have been reported and it has become a public health issue. Most symptomatic EV71 infections commonly result in HFMD or herpangina. These clinical manifestations can be associated with neurologic syndromes frequently. Neurologic syndromes observed in EV71 include meningitis, meningoencephalomyelitis, poliomyelitis-like paralytic disease, Guillain-Barr$\acute{e}$ syndrome, transverse myelitis, cerebellar ataxia, opsoclonus-myoclonus syndrome, benign intracranial hypertension, and brainstem encephalitis. Examinations for EV 71 were performed from the stools, respiratory secretion or CSF of the children by realtime PCR. Gene analysis showed that most of them were caused by EV71 subgenotype C4a which was prevalent in China, 2008. Public health measures including personal and environmental hygiene, must to target daycare centers, kindergartens, and schools where highly susceptible children congregate. To prevent the spread of infection, preschools where transmission persists for more than 2 incubation periods, have been recommended for closure, and trigger criteria for voluntary closure was instituted. During closure, operators are to thoroughly clean the centers before they are allowed to reopen. In addition, parents are advised to ensure that their children adopt a high standard of personal hygiene and to keep the infected child at home until full recovery. Because the outbreaks occur in a cyclical pattern, surveillance system to predict next outbreaks and adequate public health measures to control need to be planned for future. Control of EV71 epidemics through surveillance and public health intervention needs to be maintained in Korea. Future research should focus on understanding of EV71 virulence, identification of the receptor(s) for EV71, development of antiviral agents and development of vaccine.

• International Journal of Industrial Entomology and Biomaterials
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• 제26권1호
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• pp.31-40
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• 2013

Paralytic peptide binding proteins (PP-BP) are 30KP proteins that show similarity to ENF binding proteins. The ENF-BP act as active regulators of ENF peptides. ENF peptides are multifunctional insect cytokines. The comparison of gene expression in diapause induced and non-diapause eggs at different time intervals after oviposition showed an upregulation of PP at 18h as well as PP-BP at 12 and 18h after oviposition along with few other genes. The current study has been taken up to investigate the role of PP as well as PP-BP in diapause induction in polyvoltine silkworms and to study the multigene organization of PP-BP in the Bombyx mori genome. The tissue specific expression analysis revealed that, PP-BP is highly expressed in fat body followed by egg and brain while no expression was observed in midgut. The expression levels of PP and PP-BP in diapause and non-diapause eggs from 0h to 48h after oviposition, validated through realtime PCR revealed that PP is highly expressed at 18 and 24h while PP-BP expression is higher at 12 and 18h time intervals suggesting their possible role in diapause induction. The whole genome survey of the PP-BP paralogous sequences revealed a total of 46 B. mori PP-BP homologs that are classified into 3 categories viz., ENF-BP, Typical 30KPs and serine/threonine rich 30KPs. These paralogous sequences are distributed on chromosomes 7, 20, 22 and 24, all 30KP and S/T rich 30KP proteins are present in the same locus of chromosome 20.

• International Journal of Oral Biology
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• 제33권3호
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• pp.77-82
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• 2008

Innate immune response is initiated by the recognition of unique microbial molecular patterns through pattern recognition receptors (PRRs). The purpose of this study is to dissect the expression of various PRRs in gingival epithelial cells of differentiated versus undifferentiated states. Differentiation of immortalized human gingival epithelial HOK-16B cells was induced by culture in the presence of high $Ca^{2+}$ at increased cell density. The expression levels of various PRRs in HOK-16B cells were examined by realtime reverse transcription polymerase chain reaction (RTPCR) and flow cytometry. In addition, the expression of human beta defensins (HBDs) was examined by real time RT-PCR and the amounts of secreted cytokines were measured by enzyme linked immunosorbent assay. In undifferentiated HOK-16B cells, NACHT-LRR-PYDcontaining protein (NALP) 2 was expressed most abundantly, and toll like receptor (TLR) 2, TLR4, nucleotide-binding oligomerization domain (NOD) 1, and NOD2 were expressed in substantial levels. However, TLR3, TLR7, TLR8, TLR9, ICE protease-activating factor (IPAF), and NALP6 were hardly expressed. In differentiated cells, the levels of NOD2, NALP2, and TLR4 were different from those in undifferentiated cells at RNA but not at protein levels. Interestingly, differentiated cells expressed the increased levels of HBD-1 and -3 but secreted reduced amount of IL-8. In conclusion, the repertoire of PRRs expressed by gingival epithelial cells is limited, and undifferentiated and differentiated cells express similar levels of PRRs.