• Journal of Acupuncture Research
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• 제20권3호
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• pp.15-23
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• 2003

Objective : The role of high frequency 120 Hz electroacupuncture(EA) in carrageenan-induced pain was studied by examining the alnalgesic effects, and prostaglandin $E_2(PGE_2)$ levels measurement and spinal N-methyl-D-aspartate(NMDA) receptor expression. Inflammation was induced by an intraplantar injection of 1% carrageenan into the right hind paw. Method : Bilateral EA stimulation with 120 Hz were delivered at those acupoints corresponding to Zusanli and Sanyinjiao in man via the needles for a total of 30 min duration in carrageenan-injected rats. Results : EA stimulation showed significant analgesic effects as measured by analgesy-meter at all time points tested compared with controls. Three hours after carrageenan injection, PGE2 levels were measured by commercial kit. EA significantly inhibited PGE2 production in the right paw. The number of NR1 and NR2A, NMDA receptor, immunoreactive neurons was significantly increased in the superficial dorsal horn(laminae I-II) and nucleus proprius(laminae III-IV) of ipsilateral spinal cord at L4-5. But the number of carrageenan-induced NR1 and NR2A immunoreactive neuron, especially NR1 immunoreaction in the superficial dorsal horn, was reduced by 120 Hz EA stimulation. Conclusions : These results indicate that NMDA receptors may mediate transmission of nociceptive information originating in tissue inflammation of hind paw and high frequency 120 Hz EA stimulation have an alleviating action against local inflammatory pain.

• Journal of Acupuncture Research
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• 제20권3호
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• pp.177-193
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• 2003

Objective : The present studies investigated the effects of 120Hz high frequency electroacupunctue(EA) on the stress-induced stomach dysfunction in relation to its effect on the level of stress hormone and gastric mucosal damages. The gastric mucosal injury was induced by cold-restraint stress and two acupoints corresponding to Zusanli and Sanyinjiao in man were used. Methods: Cold-restraint stress produced typical gastric lesions in all rats of the stressed groups, but he number of ulcers as well as the mean ulcer diameter were reduced by 120 Hz EA pre-treatment. The cold-restraint stress also induced an increase in catecholamine response involving epinephrine, norepinephrine and dopamine, but an slight decline were observed in EA pre-treated rats compared with cold-restrained rats. Results: The degranulation value of gastric mast cell was significantly higher in cold-restrained rats than in control ones. However, with the significant reduction of degranulation values of gastric mast cells in EA pre-treated rats compared with cold-restrained ones, $PGE_2$ content in the gastric mucosa of EA pre-treated rats was also different from that observed in cold-restrained rats. Cold-restraint stress induced an elevated mRNA expression of pro-inflammatory gene such as cyclooxygenases-2 and tumor necrosis factor(TNF)-${\alpha}$, but these expression were down-regulated in EA pre-treated rats. Immunohistochemecal analysis showed that while the inhibitory-${\kappa}B{\alpha}$ an TNF-${\alpha}$ immunoreaction in the surface epithelium of the stomach tended to increase, both reactions in the EA pre-treated rats showed similar pattern as observed in controls. Conclusions : These results suggest that 120 Hz EA may act as a therapeutical means for gastric mucosal damages through a activation of pituitary adrenal system. it could be concluded that 120 hz high frequency electroacupuncture affords a good protective potential against stress-induce gastrointestinal dysfunction.

• 동의생리병리학회지
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• 제18권1호
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• pp.114-121
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• 2004

We investigated the effects of healthful decoction utilizing Phellinus linteus(HDPL) for suppression in the process of carbon tetrachloride (CCl₄4)-induced inflammation(50% CCl₄ : olive oil=1:1, 1 ㎖/KgㆍB.W.) of rat using biochemical, Western, histological and immunohistochemical analysis. Biochemical analysis of serum showed that the level of aspartate aminotransferase, alanine aminotransferase, lactate Dehydrogenase, alkaline phosphatase and triglyceride were significantly decreased by pretreatment of HDPL, but albumin and nitric oxide were increased. Immunoblot analysis of the liver showed that CCl₄-induced expression of interleukin-1β(IL-1β) and inducible nitric oxide synthase(iNOS) was inhibited by pretreatment of HDPL. More severe histopathological changes of the liver such as Kupffer cell reaction, inflammatory cell infiltration and focal necrosis were demonstrated in the rats challenged with CCl₄ compared with normal. Fewer scores of these changes were observed in the HDPL pretreated rats. Immunohistochemical analysis of the liver showed that while the expression of IL-1β, iNOS, tumor necrosis factor-α, COX(cyclooxygenase)-1 and COX-2 tended to increase, a decline of these immunoreaction of HDPL pre-treated groups were observed in the hepatocytes, especially in the focal necrotic sites. These results suggest that HDPL may act as a therapeutic agent for liver disease through a regulation of inflammation-related proteins.

• 동의생리병리학회지
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• 제19권6호
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• pp.1568-1572
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• 2005

Polycyclic aromatic hydrocarbon (PAH), such as benzo(a)pyrene (B(a)P), are toxic environmental contaminants known to enhance oxidative stress, production of pro-inflammatory and inflammatory cytokines. The present study was designed in order to determine whether wild ginseng (Panax ginseng C. A. Meyer) protect PAH-induced oxidative stress and inflammation. B(a)P (0.5 mg/kg, i.p.) treatment increased the distribution of immunoreactive cells for tumor necrosis factor $(TNF)-\alpha$ and cyclooxygenase (COX)-2 in peri-portal triad region and immunoreaction was shown in the cytoplasm of macrophage. Pre-treatment with wild ginseng significantly decreased immune responses in the rats treated with B(a)p. The rats given 50 mg/kg/day for 4 weeks before B(a)P treatment had 1.39-fold and 1.5-fold inhibition of $TNF-\alpha$ and COX-2 positive reaction, respectively. Wild ginseng extract alone had no effect on the distributional changes. The SOD activity as scavenger enzymes after wild ginseng administration dose-dependantly increased compared with butylated hydroxytoluene, a general radical scavenger. These data likely indicate that wild ginseng extract may act as inflammatory regulator in conjunction with inhibition of oxidant dependent metabolic activation in environmental contaminants-induced hepatic inflammation.

• 전자공학회논문지SC
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• 제48권4호
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• pp.102-107
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• 2011

본 연구에서는 복수의 항원-항체 결합 반응을 동시에 검출할 수 있는 미세유체역학 기반의 바이오칩을 설계하고 구현하였다. 본 연구의 바이오칩은 항원-항체 결합 반응이 이루어지는 반응기가 단일 채널에 직렬로 연결된 구조를 가지며, 각각의 반응기에는 항체가 고정화된 마이크로비드가 채워진다. 마이크로비드의 누출을 방지하기 위해서 마이크로채널에 위어 구조를 형성하였으며, 이를 위해서 gray-scale photolithography를 이용하였다. 항원-항체 결합 반응 검출 실험을 위해 3종의 항체를 선정하였으며, 각각의 항체를 avidn-biotin 반응을 통해 마이크로비드에 고정화하였다. 그리고, 형광물질이 표지된 항원을 마이크로채널에 연속적으로 주입하여 항원-항체 결합 반응을 유발하였으며, 10분 이내에 반응이 완료되는 것을 확인하였다. 또한, 항원에 따른 해당 반응기에서의 형광강도 증가를 검출함으로써, 미세유체 어레이의 구현 가능성을 확인하였다. 본 연구에서 제안한 미세유체 바이오칩은 면역 반응의 동시 검출을 위해 소요되는 시료의 양을 줄이고 반응 속도를 향상시킬 수 있을 것으로 사료된다.

• Journal of Microbiology and Biotechnology
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• 제29권3호
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• pp.482-488
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• 2019

Porcine circovirus type 2 (PCV2) is the causative agent of postweaning multisystemic wasting syndrome (PMWS) in pigs. Replicase (Rep) proteins are considered essential for viral replication. Capsid (Cap) protein is the primary immunogenic protein that induces protective immunity. Little is known about comparison on the immunogenicity of PCV2 Rep and Cap fusion protein and Cap protein. In the present study, recombinant baculoviruses expressing the Rep-Cap fusion protein (Bac-Rep-Cap) and the Cap protein (Bac-Cap) of PCV2 were constructed and confirmed with western blot and indirect fluorescence assay. Immunogenicities of the two recombinant proteins were tested in mice. The titers of antibodies were determined with a PCV2-specific enzyme-linked immunosorbent assay (ELISA) and a serum neutralization assay. The $IFN-{\gamma}$ response of immunized mice was measured by ELISA. The mice immunized with the Bac-Rep-Cap and Bac-Cap successfully produced Cap-specific immunoreaction. The mice immunized with the Bac-Cap developed higher PCV2-specific neutralizing antibody titers than mice injected with the Bac-Rep-Cap. $IFN-{\gamma}$ in the Bac-Rep-Cap group was increased compared to those in the Bac-Cap group. Vaccination of mice with the Bac-Rep-Cap showed significantly decreased protective efficacy compared to the Bac-Cap. Our findings will indubitably not only lead to a better understanding of the immunogenicity of PCV2, but also improved vaccines.

• 한국가시화정보학회지
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• 제20권3호
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• pp.136-145
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• 2022

Later flow immunoassay (LFIA) is a protein analytical method based on immunoreaction. On the LFIA based protein analytical method, bioreceptor molecule plays a key role, and so a system that evaluates and manages the binding affinity of bioreceptor is needed to secure detection reliability. In this study, Lateral Flow Immunoassay based rapid Bioreceptor Screening Method (rBSM) is presented that provide a simple and quick evaluating method for the binding affinity to the target protein of the antibody as model bioreceptor. To verify this evaluation method, Virus-like particles (VLP) and anti-VLP antibodies are selected as a model norovirus, which is target protein, and the candidate bioreceptors respectively. Among the 5 different candidate antibodies, appropriate antibody could be sorted out within 30 minutes through rBSM. In addition, selected antibodies were applied to two representative LFIA based techniques, sandwich assay and competitive assay. Among these methods, sandwich assay showed more effective VLP detection method. Through applying selected antibodies and techniques to the commercialized mass production lines, an VLP detecting LFIA kit was developed with a detection limit of 10¹² copies/g of VLPs in real samples. Since this proposed method in this study could be easily transformable into other combinations with bioreceptors, it is expected that this technique would be applied to LFIA kit development system and bioreceptor quality management.

• 생명과학회지
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• 제24권2호
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• pp.186-190
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• 2014

본 논문에서는 삼세기(Shaggy sea raven, Hemitripterus villosus)를 실험 재료로 선택하여 Carbonic Anhydrase Isozymes (CAs) 중의 하나인 CA III에 대한 연구를 SDS-PAGE, Isoelectric Focusing (IEF), Western blot analysis의 방법을 통하여 진행하였다. SDS-PAGE와 Western blot 결과 삼세기 아가미, 혈액, 장, 간, 신당, 근육, 심장조직에서 CA III의 분자량인 30 kDa의 band가 확인되었다. 삼세기의 근육과 아가미에 대한 등전점 전기영동(IEF)과 Western blot analysis 결과 pI 7.0 부근에서 band가 형성되는 것을 확인할 수 있었다. 특히 SDS-PAGE와 IEF 실험결과 삼세기의 아가미 조직에서 CA III의 발현량이 다른 조직들에 비하여 우세하게 나타났다. 이는 아가미가 다른 조직들과 달리 어류의 생체기관 중 유일하게 외부와 직접 접촉이 가능한 조직으로서 활성산소에 대한 손상을 최소화하기 위한 것으로 사료된다.

• 한국어병학회지
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• 제24권1호
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• pp.11-18
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• 2011

본 연구는 여윔증상을 보이는 넙치의 병태생리학적 변화를 관찰하는 것으로 목적으로 하였다. 여윔넙치가 발생한 수조 내의 정상넙치와 여윔넙치의 혈장 삼투압을 측정한 결과, 정상적인 해산어의 삼투압(280~330 mOsm/L)보다 훨씬 높은 약 361과 405 mOsm/L로 각각 나타났다. 이러한 사실은 여윔넙치와 동일한 수조 내의 정상넙치의 삼투압 조절기능에 이상이 있음을 시사하고 있다. 또한, 외관상 증상을 나타내고 있지 않으나, 여윔넙치가 발생한 수조의 정상적인 넙치는 여윔증상이 나타나기 이전의 초기단계로 정의할 수 있다. H&E염색을 통하여 여윔넙치의 장을 관찰한 결과, 여윔넙치의 전장에서는 상피세포가 남아 있기는 하나, 점막하 조직에서는 염증반응이 확인되었으며, 전장의 영양물질 흡수 기능에 손상이 야기되어 있을 가능성이 시사되었다. 그리고 후장의 경우 점막상피는 거의 손실되었으며, 직장에서는 점막상피의 소실과 아울러 소화관 고유의 folding 구조가 거의 확인되지 않았다. 면역염색의 결과에서도 장의 상피조직이 손실됨으로써 점막상피에 존재하는 $Na^+/K^+/2Cl^-$ (NKCC) cotransporter 에 대한 면역반응이 확인되지 않았다. 따라서, 해산어의 수분흡수의 중요한 장소인 후장과 직장의 상피조직이 심하게 손실됨으로써 넙치의 삼투조절기능의 균형이 깨지고 탈수현상이 보이고 따라서 여윔증상이 나타난 것으로 여겨진다. 뿐만 아니라 전장의 조직학적 변화를 통한 여윔증상에 대한 영양학적 관점에서의 고찰이 이루어져야 한다. 양식넙치의 여윔증 원인분석을 위해서 여윔증상 개체뿐만 아니라 여윔증상을 보이다가 회복된 개체에 대한 검토도 필요하다.

• 비파괴검사학회지
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• 제31권5호
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• pp.487-493
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• 2011

비표지 방식의 압전 마이크로캔틸레버 질량센서를 이용하여 간세포암의 혈청표지물질인 알파태아단백(alpha fetoprotein, AFP)을 정상 농도인 10 ng/ml까지 정량적으로 검출하였다. 압전 마이크로캔틸레버 질량센서는 캔틸레버의 질량변화에 의하여 센서의 공진주파수가 변화되는 원리를 이용하여 센서에 붙은 물질의 질량을 측정하며, 센서의 공진주파수는 컨덕턴스 스펙트럼을 이용하여 전기적으로 측정한다. 제작된 센서는 공진 주파수가 약 1.34 MHz, 질량 민감도가 약 175 Hz/pg이며 단백질이 붙을 때 캔틸레버의 표면 스트레스 변화에 대한 센서의 공진 주파수 변화를 줄일 수 있도록 설계되어 질량센서로써 신뢰도를 높였다. 'Dip and dry' 방법으로 캔틸레버의 프로브 영역을 시약과 반응시켜서 AFP 항체를 고정화하고 AFP 항원을 검출하는 실험을 수행하였다. 10 ng/ml과 50 ng/ml농도의 항원에서 10분간 항원-항체 면역반응을 시켰을 때 센서에 검출된 항원의 질량이 각각 6.02 pg과 10.67 pg이다.