Methotrexate 치료를 받고 있는 활동성 류마티스 관절염 환자에서 etanercept의 유효성과 안전성에 관한 연구

최병렬,강태영,정청일,이혜순,엄완식,김태환,전재범,유대현,배상철 대한내과학회 2004 대한내과학회지 Vol.66 No.5

목적 : 류마티스 관절염을 앓고 있는 한국인 중에 기존 DMARDs로 치료 실패하였고, 고정용량의 MTX를 복용하고 있는 환자들에 있어서 etanercept의 유효성과 안전성을 평가하고자 하였다. 방법 : 기존 DMARDs로 치료 실패한 활동성 류마티스 관절염 환자 76명을 대상으로 하여 단일군, 공개시험을 하였다. 대상 환자들은 고정용량의 MTX를 복용하면서 etanercept 25 mg을 1주일에 두 번 피하 주사하였으며 12주간 투여하였다. 유효성은 ACR 20, ACR 50,조조강직 시간으로 평가하였고, 약제의 안전성은 이상반응 등으로 평가하였다. 결과 : 대상 환자는 총 76명으로 평균 연령은 45.2세, 남자 5명, 여자 71명이었다. 84.4%인 54명이 12주째에 ACR 20을 만족하였고, 53.1%인 34명이 12주째에 ACR 50을 만족하였다. 조조 강직 시간은 치료 전 203.3분에서 치료 12주째 42.6분을 평균 74.5% 호전되었다. 가장 흔한 이상반응은 주사부위 반응이었다. 이외에도 상기도 감염, 오심, 안면부종 등이 발생하였으나 심각한 부작용은 없었다. 결론 : etanercept는 효과적이고, 안전한 류마티스 관절염 치료 방법이며 특히 MTX치료에도 불구하고 활동성인 류마티스 관절염에 기대되는 치료라고 할 수 있다. Background : This study was performed to investigate the efficacy and safety of etanercept in active rheumatoid arthritis patients with stable dose of methotrexate in Korean. Methods : In a 12 week, single arm, open trial, we assigned 76 patients with active rheumatoid arthritis who had an inadequate response to disease-modifying antirheumatic drugs. Patients received twice-weekly subcutaneous injections of etanercept 25 ㎎ while containing to receive methotrexate at a stable dose of 7.5~25 ㎎ per week. The clinical response was defined as the percent improvement in disease activity according to the criteria of the American Collage of Rheumatology (ACR) at 12 weeks. Results : Etanrecept led to significant improvements in disease activity and was safe and well tolerated. At 12 week, 84.4% of the patients receiving 25 ㎎ of etanercept achieved a 20% ACR response, and 53.1% of those receiving etanercept achieved a 50% ACR response. The most common adverse event was injection-site reaction. Other advanse events were upper respiratory infection, nausea, and facial edema, but there were no serious adverse events associated with etanercept. Conclusion : In active rheumatoid arthritis patients, etanercept was safe, well tolerated, and provided rapid clinical improvements.

제조합 대장균에서 과발현된 Citrobacter freundii KCTC2006 유래의 β-Tyrosinase를 이용한 3,4-Dihydroxyphenyl-L-alanine의 생산

이승구,노현수,홍승표,이규종,왕지원,태동년,엄기남,방상구,김영준,성문희 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.1

재조합 대장균에서 대량발현 시킨 Citrobacter freundii KCTC 2006 유래이 효소 β-tyrosinase를 이용하여 pyrocatechol, sodium pyruvate, ammonium acetate로부터 3,4-dihydroxy phenyl-L-alanine을 생산하기 위한 연구를 수행하였다. 이 효소반응에 적합한 온도 및 pH 조건은 각각 18℃와 8.5로 결정되었고, 반응액 중의 ammonium acetate와 sodium pyruvate의 농도는 각각 300 mM, 50 mM 이상으로 조절하는 것이 적합하였다. Pyrocatechol의 경우는 20 mM에서 가장 높은 반응성을 나타냈으나, 기질을 반복적으로 첨가하며 장시간 동안 효소반응을 수행하는 경우에는 pyrocatechol의 고갈을 피하기 위하여, 20 mM에서 50 mM 사이로 조절하였다. 한편, 반응액 중에 ethanol을 10% 첨가한 경우에는 반응속도가약 20% 증가하였다. 이상과 같은 효소반응특성에 기초하여 조제한 기질용액에 β-tyrosinase를 1 unit/㎖ 농도로 가하고, pyrocatechol과 pyruvate가 고갈되지 않도록 간헐적으로 첨가하면서 효소반응을 수행한 결과, 24시간 만에 85.2%의 수율로 31.6g/ℓ의 3,4-dihydroxyphenyl-L-alanine를 생산할 수 있었다. By using the β-tyrosinase of Citrobacter freundii KCTC2006, which was cloned and overexpressed in Escherichia coli, 3,4-dihydroxy phenyl-L-alanine (L-DOPA) was synthesized efficiently from pyrocatechol, sodium pyruvate, and ammonium acetate. Optimal temperature and pH for the reaction were determined to be about 18℃ and 8.5, respectively. The effects of substrate concentrations were also examined at different concentrations of ammonium acetate, sodium pyruvate, and pyrocatechol. Ammonium acetate and sodium pyruvate increased the reaction rate until the concentrations reached to 300 mM and 50 mM, respectively. Although pyrocatechol showed the optimal concentration at 20 mM, it was controlled between 20 mM and 50 mM to avoid the depletion of substrate during the enzymatic synthesis. Based on above results, a reaction medium for the production of L-DOPA was prepared and incubated with 1 unit/㎖ of β-tyrosinase. Pyrocatechol and sodium pyruvate was added to the reaction solution intermittently to avoid the substrate depletion during the enzymatic reaction. After 24 hour of reaction, 31.6 g/ℓ L-DOPA was accumulated in the reaction solution as soluble and precipitated ones and the conversion yield was about 85.2%.

Developmental Ability of Porcine Fragmented Parthenote and Cloned Embryos Produced by Somatic Cell Nuclear Transfer

Uhm, Sang-Jun,Gupta, Mukesh Kumar,Kim, Min-Seok,Park, Chan-Kyu,Chung, Kil-Saeng,Lee, Hoon-Taek 한국수정란이식학회 2005 한국수정란이식학회 학술대회 Vol.2005 No.1

Effect of Epidermal Growth Factor on the Fertilization and Development of In Vitro - Matured Porcine Oocytes

Sang Jun Uhm,Eun Young Kim,Myo Kyung Kim,San Hyun Yoon,Se Pill Park,Kil Saeng Chung,Jin Ho Lim 한국동물자원과학회(구 한국축산학회) 1996 한국축산학회 심포지움자료집 Vol.- No.-

The Effects of NaCl and SIT on the Development of IVM - IVF Porcine Embryos

Sang Jun Uhm,Se Pill Park,Seun Eui Kim,San Hyun Yoon,Jin Ho Lim 한국동물자원과학회(구 한국축산학회) 1995 한국축산학회 심포지움자료집 Vol.- No.-

Relationship between Developmental Ability and Cell Number of Day 2 Porcine Embryos Produced by Parthenogenesis or Somatic Cell Nuclear Transfer

Uhm, Sang Jun,Gupta, Mukesh Kumar,Chung, Hak-Jae,Kim, Jin Hoi,Park, Chankyu,Lee, Hoon Taek Asian Australasian Association of Animal Productio 2009 Animal Bioscience Vol.22 No.4

In vitro produced porcine embryos have potential application in reproductive biotechnology. However, their development potential has been very low. This study evaluated the in vitro developmental ability and quality of cloned and parthenogenetic porcine embryos having 2-4 cells or 5-8 cells on Day 2 of in vitro culture. Analysis of results showed that 2 to 4 cell embryos had higher ability to form blastocysts than 5 to 8 cell embryos (p<0.05). Blastocysts produced from culture of 2 to 4 cell embryos also contained higher cell numbers and had lower BAX:BCLxL transcript ratio than those produced from 5 to 8 cell embryos (p<0.05), thereby suggesting 2 to 4 cell embryos have higher development potential. Further investigation revealed that 5 to 8 cell embryos had higher incidence (100${\pm}$0.0%) of blastomeric fragmentation than 2 to 4 cell embryos (15.2${\pm}$5.5% for parthenogenetic and 27.7${\pm}$7.1% for cloned embryos). This suggests that low development potential of 5 to 8 cell embryos was associated with blastomeric fragmentation. In conclusion, we have shown that morphological selection of embryos based on cell number on Day 2 of in vitro culture could offer a practical and valuable non-invasive means to select good quality porcine embryos.

3-Hydroxyflavone Improves the <i>In Vitro</i> Development of Cloned Porcine Embryos by Inhibiting ROS Production

Uhm, Sang Jun,Gupta, Mukesh Kumar,Das, Ziban Chandra,Kim, Nam-Hyung,Lee, Hoon Taek Mary Ann Liebert 2011 Cellular reprogramming Vol.13 No.5

<P>UV-irradiation of oocytes during enucleation and serum starvation of donor cells during cell cycle synchronization may compromise the development competence of cloned embryos through excessive generation of reactive oxygen species (ROS). Here, we show that 3-hyroxyflavone (a flavonoid having hydroxyl group at 3 carbon position) inhibits UV- and serum starvation-induced ROS production in oocytes and donor cells, respectively, and thereby improves the in vitro development of cloned porcine embryos (p<0.05). In a parthenogenetic model, UV-irradiation for 5?sec or more was found to reduce the in vitro development and quality of the embryo, which could be rescued by their culture in the presence of 3-hydroxyflavone. The rescuing effect of 3-hydroxyflavone was associated with significant reduction in ROS level (14.41.0 vs. 47.16.7), increase in ERK signaling molecules by 2.1-fold, and decrease in Caspase3 expression by 3.2-fold. Culture of donor cells (18.51.4 vs. 13.01.7%) or cloned embryos (20.61.1 vs. 12.21.1%) in the presence of 3-hydroxflavone also increased (p<0.05) the rates of blastocyst formation in cloned embryos produced by the nuclear transfer of serum-starved donor cells into recipient cytoplasts exposed to UV-irradiation during the enucleation step. Importantly, both parthenotes and cloned embryos cultured in the presence of 3-hydroxyflavone had significantly increased ability to expand, and contained a higher number of cells than those of the control group (p<0.05). These results suggest that 3-hydroxyflavone may be useful for improving the in vitro developmental potential of cloned embryos through inhibition of ROS production induced by the UV-irradiation of oocyte and/or the serum starvation of donor cells.</P>

Effect of supplement of SCM in culture medium for in vitro development of bovine in vitro fertilized oocytes

Sang Jun Uhm The Korean Society of Animal Reproduction and Biot 2023 한국동물생명공학회지 Vol.38 No.3

Background: The successful production of superior or transgenic offspring from in vitro produced embryos in cattle relies heavily on the quality of blastocyst stage embryos. In order to enhance the developmental competency of these embryos, a novel culture method was devised. Methods: This study utilized stem cell culture medium (SCM) from hESCs as a supplement within the culture medium for bovine in vitro produced embryos. To gauge the efficacy of this approach, in vitro fertilized embryos were subjected to culture in CR1aa medium enriched with one of three supplements: 0.3% BSA, 10% FBS, or 10% SCM. Results: The blastocyst development and hatching rates of one-cell zygotes cultured in CR1aa medium supplemented with SCM (23.9% and 10.2%) surpassed those cultured in CR1aa medium supplemented with BSA (9.3% and 0.0%) or FBS (3.1% and 0.0%) (p < 0.05). Furthermore, post-zygotic gene activation, cleaved embryos cultured in CR1aa medium supplemented with SCM (57.8% and 34.5%) exhibited notably higher rates (p < 0.05) compared to those cultured with BSA (12.9% and 0.0%) or FBS (45.7% and 22.5%) supplementation. Furthermore, the microinjection of SCM into the cytoplasm or pronucleus of fertilized zygotes resulted in elevated blastocyst development and hatching rates, particularly when the microinjected embryos were subsequently cultured in CR1aa medium supplemented with SCM from the 8-cell embryo stage onwards (p < 0.05), in contrast to those cultured with FBS supplementation. Conclusions: In conclusion, this study conclusively demonstrated that the incorporation of SCM into the culture medium significantly enhances the developmental progress of preimplantation embryos.

Expression of enhanced green fluorescent protein in porcine- and bovine-cloned embryos following interspecies somatic cell nuclear transfer of fibroblasts transfected by retrovirus vector

Uhm, Sang Jun,Gupta, Mukesh Kumar,Kim, Teoan,Lee, Hoon Taek JOHN WILEY & SONS LTD 2007 MOLECULAR REPRODUCTION AND DEVELOPMENT Vol.74 No.12

<P>Interspecies somatic cell nuclear transfer (iSCNT) has emerged as an important tool for studying nucleo-cytoplasmic interactions and cloning of animals whose oocytes are difficult to obtain. This study was designed to explore the feasibility of employing transgenic fibroblasts as donor cells for iSCNT. The study examined the chromatin morphology, in vitro development, and expression of an enhanced green fluorescent protein (EGFP) gene in porcine- and bovine-cloned embryos produced by iSCNT of fetal fibroblast transfected with a pLNβ-EGFP retroviral vector. Parthenogenetic and transfected or nontransfected intraspecies SCNT embryos were used as controls for comparison. Analysis of data revealed that xenogenic oocyte was able to reprogram somatic cells of different genus and supports their in vitro development to the blastocyst stage. However, the developmental rates of transgenic iSCNT embryos to the blastocyst stage were significantly lower than those of intraspecies SCNT embryos. The reduction in development rates was however, not due to integration of the transgene as the lower (P < 0.05) development rates of the intraspecies SCNT porcine or bovine embryos did not differ between transgenic and nontransgenic groups. Expression of EGFP was observed in 100% of blastocysts and mosaicism was not observed. Furthermore, after iSCNT of porcine or bovine donor nuclei into xenogenic ooplasm, patterns of nuclear remodeling in reconstructed embryos were similar. In conclusion, our data demonstrated the feasibility of producing transgenic iSCNT embryos. To our knowledge, this is the first report of transgenic cloned embryo production by iSCNT approach. In the future, this may provide a powerful research tool for studying developmental events in domestic animals and provide marked cell lines for other genetic manipulations. Mol. Reprod. Dev. 74: 1538–1547, 2007. © 2007 Wiley-Liss, Inc.</P>

Developmental Ability of Porcine Fragmented Parthenote and Cloned Embryos Produced by Somatic Cell Nuclear Transfer

Uhm Sang-Jun,Gupta Mukesh Kumar,Kim Min-Seok,Park Chan-Kyu,Chung Kil-Saeng,Lee Hoon-Taek 한국발생생물학회 2005 한국발생생물학회 학술발표대회 Vol.2005 No.1