• 대한약학회:학술대회논문집
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• 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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• pp.171.2-172
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• 2002

• Pediatric Gastroenterology, Hepatology & Nutrition
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• 제2권2호
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• pp.169-177
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• 1999

목 적: 소아 만성 B형 간염 환아에서 알파 인터페론의 치료 효과는 보고자마다 차이가 있으나 30~40%로 알려져 있다. 그러나 만성 지속성 B형 간염, 혈청 ALT치가 낮은 경우나 혈청 HBV DNA치가 높은 소아 만성 B형 간염 환자에서 알파 인터페론 단독 투여 시 그 치료 효과는 더 낮다고 보고되고 있다. 이에 저자들은 알파 인터페론 단독 투여 시 치료 효과가 낮다고 알려진 소아 만성 B형간염 환자에 prednisolone 이탈 요법 후 인터페론 병용 투여 시 그 치료 효과를 알아보고자 본 연구를 시행하였다. 대상 및 방법: 1996년 1월부터 1997년 12월까지 전남대학교병원 소아과에 내원하여 만성 B형 간염으로 진단받은 28명을 대상으로 하였다. 대상 환아 중 간 조직 검사 상 만성 활동성 간염을 보이고 혈청 ALT치가 100 IU/L 이상이고 혈청 HBV-DNA치가 100 pg/$300\;{\mu}L$ 미만인 14명(group 1)은 알파 인터페론을 체표면적($m^2$) 당 5백만 단위를 6개월 동안 주 3회 단독 투여하였다. 조직 검사 상 만성 지속성 간염인 경우와 만성 활동성 간염이면서 혈청ALT치가 100 IU/L 미만 또는 혈청 HBV DNA치가 100 pg/$300\;{\mu}L$ 초과한 14명(group 2)은 prednisolone을 60 mg/$m^2$, 40 mg/$m^2$, 20 mg/$m^2$으로 각각 2주씩 6주간 경구 투여하고 2주간 휴약 기간을 가진 후 알파 인터페론을 group 1과 같은 방법으로 투여하였다. 치료에 대한 반응은 인터페론 투여가 종료되는 시점에서 완전반응(항 HBe 항체의 양전화, 혈청 ALT 정상화 및 혈청 HBV-DNA 음성), 부분반응(혈청 ALT 정상화 또는 혈청 HBV-DNA 음성) 및 무반응으로 평가하였다. 결 과: 1) 대상 환아의 평균 연령은 130.6개월(21~192개월)이었고, 남아는 22례, 여아는 6례이었다. 만성 지속성 간염은 11례, 만성 활동성 간염은 17례이었고, 15명의 환아에서 모친이 B형 간염 바이러스 만성 보유자이었다. 2) Group 1에서 평균 연령은 144.1개월(97~169개월), 남아 9명, 여아 5명, 혈청 ALT $299.9{\pm}215.3$ IU/L, HBV-DNA $49.3{\pm}33.1\;pg/300\;{\mu}L$이었고, group 2에서 평균 연령은 112.1개월(21~192개월), 남아 13명, 여아 1명, 만성 지속성 간염 11명, 만성 활동성 간염 3명, 혈청 ALT $85.6{\pm}71.0$ IU/L, HBVDNA $524.3{\pm}1064\;pg/300\;{\mu}L$이었다. 3) Group 1에서 항 HBe 항체 양전은 10례(71.4%) 에서, 혈청 ALT는 9례(64.3%)에서 정상화되었고, HBV-DNA는 11례(78.6%)에서 음성화되었다. Group 2에서 항 HBe 항체 양전은 7례(50.0%)에서, 비정상 수치를 보인 9명의 환아 중 5명(55.6%)이 ALT 의 정상화를 보였고, HBV-DNA는 9례(64.3%)에서 음성화되었다. 두 군간의 항 HBe 항체의 양전율, ALT치의 정상화율, HBV DNA치의 음성화율에 있어서 통계학적으로 유의한 차이는 없었다. 4) Group 1에서 완전반응은 8례(57.1%), 부분반응은 3례(21.4%), 무반응은 3례(21.4%)이었고, group 2 에서는 완전반응 7례(50.0%), 부분반응 2례(14.3%), 무반응 5례(35.7%)으로 두 군간의 통계학적 유의한 차이는 없었다. 결 론: 소아 만성 B형 간염에 대한 치료로 인터페론 단독 요법으로 반응이 좋지 않을 것으로 예측되는 환자군에 스테로이드 이탈 요법 후 인터페론 병합 투여는 안전하고 효과적인 치료법으로 생각된다. 향후 치료 효과의 지속 여부에 대한 지속적인 관찰과 더 많은 환자를 대상으로 한 전향적인 비교 연구가 필요하리라 사료된다.

• 셀메드
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• 제2권3호
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• pp.27.1-27.6
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• 2012

Red ginseng, which has a variety of biological and pharmacological activities including antioxidant, anti-inflammatory, antimutagenic and anticarcinogenic effects, has been used for thousands of years as a general tonic in traditional oriental medicine. Here, we tested the immune regulatory activities of hydrolyzed red ginseng by malted barley (HRG) on the expressions of receptor interacting proteins (Rip) 2 and $I{\kappa}B$ kinase-beta (IKK-${\beta}$) in mouse peritoneal macrophages. We show that HRG increased the activations of Rip 2 and IKK-${\beta}$ for the first time. When HRG was used in combination with recombinant interferon-${\gamma}$ (rIFN-${\gamma}$), there was a marked cooperative induction of nitric oxide (NO) production. The increased expression of inducible NO synthase from rIFN-${\gamma}$ plus HRG-stimulated cells was almost completely inhibited by pre-treatment with pyrrolidine dithiocarbamate (PDTC), an inhibitor of nuclear factor-${\kappa}B$ (NF-${\kappa}B$). In addition, the treatment of peritoneal macrophages with rIFN-${\gamma}$ plus HRG caused significant increases in tumor necrosis factor (TNF)-${\alpha}$ mRNA expression and production. Because NO and TNF-${\alpha}$ play an important role in the immune function and host defense, HRG treatment can modulate several aspects of the host defense mechanisms as a result of the stimulations of the inducible nitric oxide synthase and NF-${\kappa}B$. In conclusion, our findings demonstrate that HRG increases the productions of NO and TNF-${\alpha}$ from rIFN-${\gamma}$-primed macrophages and suggest that Rip2/IKK-${\beta}$ plays a critical role in mediating these immune regulatory effects of HRG.

• 대한한의학회지
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• 제21권3호
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• pp.20-30
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• 2000

Objective : This experiment was performed in order to study the effect of an aqueous extract of Salviae radix root(SRRAE) on NO production and NOS gene induction from macrophages Methods : To investigate dose-dependent effects of SRRAE for NO release on the $rIFN-{\gamma}-treated$ macrophages, the cells were incubated for 6 hrs in a medium containing $rIFN-{\gamma}$ (5 U/ml), stimulated with SRRAE and incubated in a CO2 incubator. The cells were treated with 5 U/ml $rIFN-{\gamma}$ plus 100 g/ml of SRRAE, Then, the cells were incubated with various concentrations of NGMMA at $37^{\circ}C$ for 48 hrs, Results : SRRAE had no effect on NO production by itself, whereas recombinant $interferon-{\gamma}(rIFN-{\gamma})$ alone showed modest activity, When SRRAE was used in combination with $rIFN-{\gamma}$, there was a marked cooperative induction of NO production in a dose-dependent manner. The optimal effect of SRRAE on NO production was shown at 6hrs after treatment with $rIFN-{\gamma}$. The SRRAE-induced production of NO was inhibited by NG-monomethyl- L-arginine(NGMMA) and arginase. $rIFN-{\gamma}$ in combination with SRRAE showed a marked increase of the expression of the inducible NOS(iNOS) gene. In addition, the effect of SRRAE was mainly dependent on the SRRAE-induced tumor necrosis $factor-{\alpha}(TNF-{\alpha})$ secretion. Conclusions : SRRAE induces NO production from macrophages as a result of SRRAE-induced $TNF-{\alpha}$ secretion. SRRAE may provide a second signal for synergistic induction of NO production in macrophages already induced to express iNOS gene by $rIFN-{\gamma}$.

• 한방안이비인후피부과학회지
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• 제11권1호
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• pp.1-22
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• 1998

During the last few years, nitric oxide(NO) as a potent macrophage-derived effector molecule against a variety of bacteria, parasites, and tumors has received increasing attention. More recent studies suggest that NO also has antiviral effects in both murine and human cells. The objective of the current study was to determine the effect of Yongdamsagantang(YST) on the production of NO. Stimulation of mouse peritoneal macrophages with YST after the treatment of recombinant $interferon-{\gammer}(rlFN-{\gammer})$ resulted in the increased NO synthesis. YST had no effect on NO synthesis by itself. When YST was used in combination with $rIFN-{\gammer}$, there was a marked cooperative induction of NO synthesis in a dose-dependent manner. The optimal effect of YST on NO synthesis was shown 6 hour after treatment with $rIFN-{\gammer}$. This increase in NO synthesis was reflected as increased amount of inducible NO synthase(iNOS) protein. NO production was inhibited by $N^G-monomethyl-L-arginine$. The increased production of NO from $rIFN-{\gammer}$ plus YST-stimulated cells was decreased by the treatment with staurosporin. In addition, synergy between $rIFN-{\gammer}$ and YST was mainly dependent on YST-induced tumor necrosis $factor-{\alpha}(TNF-{\alpha})$ secretion. These results suggest that the capacity of YST to increase NO production from $rIFN-{\alpha}-primed$ mouse peritoneal macrophages is the result of YST-induced $TNF-{\alpha}$ secretion.

• Asian-Australasian Journal of Animal Sciences
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• 제26권6호
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• pp.795-803
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• 2013

Various endometrial genes in ruminant ungulates are regulated by conceptus interferon tau (IFNT). However, the effect of each IFNT isoform has not been carefully evaluated. In this study, the effects of 2 IFNT isoforms, paralogs found in utero, and interferon alpha (IFNA) on uterine epithelial and Mardin-Darby bovine kidney (MDBK) cells were evaluated. Expression vectors of the bovine interferon (bIFNT) genes bIFNT1, bIFNTc1, and bIFNA were constructed, and recombinant bIFNs (rbIFNs) were produced by 293 cells. Bovine uterine epithelial or MDBK cells were cultured in the presence or absence of increasing concentrations of each rbIFN for 24, 48, or 72 h. Transcript levels of the IFN-stimulated genes (ISGs) ISG12, ISG15, MX1, and MX2 were analyzed using quantitative reverse transcription-polymerase chain reaction. These messenger RNAs were up-regulated by rbIFN in a time- and concentration-dependent manner. In the epithelial cells, the ISG12 transcript level increased at 48 h after rbIFN treatment but slightly decreased at 72 h, whereas the transcript level of ISG15 increased at 24 h and was maintained through 72 h. Expressions of MX1 and MX2 increased at 72 h after rbIFN treatment. MX1 expression increased in all treatment groups, but MX2 increased only by bIFNTc1. In MDBK cells, the expression of ISG12 was increased by bIFNT1 and bIFNTc1 after 24 and 72 h; however, it was unchanged by rbIFNA. ISG15 increased following the same pattern as that seen in uterine epithelial cells, and MX1 showed a similar expression pattern. MX2 expression was increased by bIFNTc1 treatment in uterine epithelial cells, and its expression was increased by both bIFNT1 and bIFNTc1 in MDBK cells. These results show that epithelial and MDBK cell responses to IFNs differ, suggesting that IFNs possess common functions, but may have acquired different functions following gene duplication.

• 대한한방내과학회지
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• 제20권1호
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• pp.210-221
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• 1999

A clinial observation was done on 12 cases of paralytic ileus patients, treated by Bo-Ryu Enema(保留灌腸), who were hospitalized from May 1, 1995 to October 31, 1996 at the Department of Oriented Internal Medicine II, Oriental Medicine Hospital. Taejon University. The results were as follows; 1. The ratio between male and female was 1 : 1.4. The distribution of age. 70' years or over, 60', 50' years generation were revealed in turn. 2. In classiffication of human coporeal constitution, Soeumin(少陰人) were 9 cases(75.0%), Taeumin(太陰人) 2 cases (16.7%), Soyangin(少陽人) 1 case (8.3%). 3. In distribution of disease on admission, Stroke sequela was the most number with 7 cases(58.3%), Stroke 3 cases(25.3%). Hypertensive encephalopathy and Brain tumor were 1 case, each other. 4. The effect of treated by Bo-Ryu Enema was as follows: Each of Excellent(良好) and Good(好戰) were 6 cases(50%) but. Fair(別無好戰) and Poor(惡化) were no case.

• 한국미생물·생명공학회지
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• 제49권1호
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• pp.75-87
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• 2021

Type I Interferons (IFNα) are known for their role as biological anticancer agents owing to their cell-apoptosis inducing properties. Development of an appropriate, cost-effective host expression system is crucial for meeting the increasing demand for proteins. Therefore, this study aims to develop codon-optimized IFNα-2b in L. lactis NZ3900. These cells express extracellular protein using the NICE system and Usp₄₅ signal peptide. To validate the mature form of the expressed protein, the recombinant IFNα-2b was screened in a human colorectal cancer cell line using the cytotoxicity assay. The IFNα-2b was successfully cloned into the pNZ8148 vector, thereby generating recombinant L. lactis pNZ8148-SP_Usp45-IFNα-2b. The computational analysis of codon-optimized IFNα-2b revealed no mutation and amino acid changes; additionally, the codon-optimized IFNα-2b showed 100% similarity with native human IFNα-2b, in the BLAST analysis. The partial size exclusion chromatography (SEC) of extracellular protein yielded a 19 kDa protein, which was further confirmed by its positive binding to anti-IFNα-2b in the western blot analysis. The crude protein and SEC-purified partial fraction showed IC₅₀ values of 33.22 ㎍/ml and 127.2 ㎍/ml, respectively, which indicated better activity than the metabolites of L. lactis NZ3900 (231.8 ㎍/ml). These values were also comparable with those of the regular anticancer drug tamoxifen (105.5 ㎍/ml). These results demonstrated L. lactis as a promising host system that functions by utilizing the pNZ8148 NICE system. Meanwhile, codon-optimized usage of the inserted gene increased the optimal protein expression levels, which could be beneficial for its large-scale production. Taken together, the recombinant L. lactis IFNα-2b is a potential alternative treatment for colorectal cancer. Furthermore, its activity was analyzed in the WiDr cell line, to assess its colorectal anticancer activities in vivo.

• KSBB Journal
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• 제21권2호
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• pp.133-139
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• 2006

혈액 내 순환시 안정성 향상과 면역원성의 감소를 위해, rhIFN-${\alpha}$-2a은 N-terminus의 ${\alpha}$-아민기에 mPEG aldehyde를 solid-phase PEGylation 시킨다. CM-Sepharose와 같은 양이온 교환수지가 고체 지지체로 사용되었다. Mono-PEGylate는 양이온 교환 수지에서 unmodified 단백질과 분리되어 용출된다. Site-srecific PEGylation과 mono-PEGylate의 분리가 한 단계의 공정으로 얻어진다는 점은 solid-phase PEGylation의 이점을 뒷받침해준다. 위치 특이성은 peptide digest의 질량 분석과 Edman degradation을 이용한 N-terminal sequencing에 의해 확인하였다. Mono-PEGylate는 항바이러스 활성과 면역원성의 감소를 나타내고, 감소 정도는 결합되는 mPEG의 분자량에 비례한다. Trypsin 저항성과 온도 안정성은 mono-PEGylation에 의해 두드러지게 개선되었다. Solid-phase PEGylation을 통해 종래의 액상 반응에서 나타날 수 있는 재현성 낮은 반응, 부 반응물 생성, 부 반응물 제거 공정 등의 단점을 극복할 수 있었다. 그러나 solid-phase PEGylation의 문제점인 액상 반응에 비교하여 많은 양의 PEG를 사용하여야 한다는 점은 개선되어야 한다.

• Toxicological Research
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• 제31권1호
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• pp.11-16
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• 2015

Our body's immune system has defense mechanisms against pathogens such as viruses and bacteria. Immune responses are primarily initiated by the activation of toll-like receptors (TLRs). In particular, TLR4 is well-characterized and is known to be activated by gram-negative bacteria and tissue damage signals. TLR4 requires myeloid differentiation factor 2 (MD2) as a co-receptor to recognize its ligand, lipopolysaccharides (LPS), which is an extracellular membrane component of gram-negative bacteria. Gambogic acid is a xanthonoid isolated from brownish or orange resin extracted from Garcinia hanburyi. Its primary effect is tumor suppression. Since inflammatory responses are related to the development of cancer, we hypothesized that gambogic acid may regulate TLR4 activation. Our results demonstrated that gambogic acid decreased the expression of pro-inflammatory cytokines ($TNF-{\alpha}$, IL-6, IL-12, and $IL-1{\beta}$) in both mRNA and protein levels in bone marrow-derived primary macrophages after stimulation with LPS. Gambogic acid did not inhibit the activation of Interferon regulatory factor 3 (IRF3) induced by TBK1 overexpression in a luciferase reporter gene assay using IFN-${\beta}$-PRD III-I-luc. An in vitro kinase assay using recombinant TBK1 revealed that gambogic acid did not directly inhibit TBK1 kinase activity, and instead suppressed the binding of LPS to MD2, as determined by an in vitro binding assay and confocal microscopy analysis. Together, our results demonstrate that gambogic acid disrupts LPS interaction with the TLR4/MD2 complex, the novel mechanism by which it suppresses TLR4 activation.