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Immunoreaction of Human Tissue Plasminogen Activator Harboring-Gene Expression in Transgenic Pig
심미란,Mi-Yun Oh,Kyong-Woon Kim,Hwi-Cheul Lee,양병철,Hee-Kyung Chung,박춘근,Jin-Ki Park,Hak-Jae Chung 사단법인 한국동물생명공학회 2011 Reproductive & developmental biology Vol.35 No.3
The aim of this study was to examine the possibility of transgenic animals as bioreactor. Transgenic animals are required to synthesize proteins with the appropriate post-translational modifications and being used for this purpose. The bladder and mammary gland from transgenic animals are important bioreactor. In the study, we used the expression plasmid DNA containing mouse uroplakin Ⅱ promoter and bovine α-S1 casein promoter. Transgenic pigs as recipient transferred to embryos containing human tPA transgene through microinjection of fertilized one cell zygotes. Thus, Human tissue-type plasminogen activactor (tPA) protein was produced to direct bladder and mammary gland by action of the bladder-specific promoter and mammary gland-specific promoter. Secretion of human tPA in bladder and mammary gland were confirmed by western blot analyses. These data suggests that the bladder and mammary gland can be used as a bioreactor to produce useful protein. Production of human tPA transgenic pig could have a very positive impact in term of cost and timescale. Furthermore, it suggests a potentional application of bladder expression system and mammary gland expression system for high-yield production of therapeutic proteins.
정학재,심미란,조은별,조유미,최명섭,오미연,정희경,김경운,한덕우,양병철,박진기,장원경,박춘근 한국동물번식학회 2012 Reproductive & Developmental Biology(Supplement) Vol.36 No.2s
유즙 내에 들어있는 유용 단백질을 분리, 정제하였을 때 예상되는 경제적 가치 때문에 우 리는 유즙 내에 들어있는 재조합 hEPO 단백질을 추출하고 정제하려는 많은 시도를 하고 있다. 이러한 노력에도 불구하고 유즙 내에 들어있는 target 단백질의 정제가 매우 어렵고, 그 순도 역시 20%에 지나지 않는다. 우리는 hEPO 유전자를 가지고 있으며 유선에서 hEPO를 분비하는 형질전환 돼지를 생산 보유하고 있다. 그러나, 이들 형질전환 돼지들은 체내에 도입된 유전자의 발현에 의해 야기되는 적혈구 과다증 그리고 혈소판 감소증과 같 은 순환기의 문제가 유발되고 있다. 명백하게, 이러한 종류의 혈액학적 변화는 장기의 정상 적인 기능을 방해하며 형질전환 돼지의 갑작스런 죽음을 초래한다. 그러므로, 앞서 언급한 문제점들을 해결하기 위한 alternative한 방법이 필요하다. 현재 연구에서, 우리는 외인성 hEPO를 발현하는 돼지에서 비정상의 생리학적 증후에 관해서 실험하였으며 hEPO를 생산 하기 위한 alternative choice로서 형질전환 돼지 유래 세포의 배양 system을 제안하였다. 5마리의 F6 hEPO 형질전환 돼지와 4마리와 대조군으로 일반돼지(랜드레이스)를 국립축 산과학원 동물바이오공학과에서 사육 도축하고 순환 장기를 채취하였다. 형질전환 돼지 순 환기 조직(유선, 신장, 폐, 비장)을 계대 배양 후 hEPO mRNA의 발현을 RT-PCR 방법으로 분석하였다. 또한, 면역조직화학 염색법을 통해서 hEPO 형질전환돼지 순환 장기조직에서 발현되는 hEPO의 발현 양상을 분석한 결과로서 유선, 신장 및 폐 조직에서 hEPO의 발현 이 확인되었으나, hEPO 발현 양상이 모든 형질전환동물 조직에서 발현되는 경향을 보이지 는 않았다. 한편, 세포면역화학 분석법에서는 형질전환돼지 유래 세포의 배양 시 hEPO가 발현되고 있는 양상을 확인하였다. 또한, 형질전환 돼지와 일반돼지의 세포 증식률과 증식 속도는 신장 세포에서는 빠른 증식을 나타낸 반면, 폐와 비장 세포에서는 느린 증식률이 확 인되었다. 배양 후 지속적인 세포 증식과 세포의 생존성을 분석하기 위하여 Apoptosis를 TUNEL과 Annexin V를 이용한 방법으로 조사한 결과, 형질전환돼지 유래의 세포와 일반돼 지의 세포 사이에 유의적 차이는 발견하지 못했다. 이들의 결과들로서 본 연구에서는 형질 전환돼지를 생산하여 유용 단백질을 이용하고자 할 경우, 목적으로 하는 세포 뿐 만 아니라 형질전환가축 유래의 다양한 세포를 이용 할 수 있는 가능성을 시사하고 있다.
활창과 허밍을 이용한 음성치료가 성문틈 환자의 음성 개선에 미치는 효과
정대용,심미란,황연신,김근전,선동일,Jung, Dae-Yong,Shim, Mi-Ran,Hwang, Yeon-Shin,Kim, Geun-Jeon,Sun, Dong-Il 대한후두음성언어의학회 2021 대한후두음성언어의학회지 Vol.32 No.2
Background and Objectives Therapies have been reported to treat the glottal gap previously. However, these voice therapies showed the limits because many techniques focused only on one among breathing, resonance and phonation. In addition patients often have difficulties visiting hospital frequently. 'Gliding and humming' is vocal training technique that readjusts total vocal patterns such as breathing, resonance and phonation. This technique can be easily applied during short term sessions. The purpose of this study is to evaluate the efficiency of voice therapy with 'gliding and humming' for patients with glottic gap during short-term treatment sessions. Materials and Method Twenty-three patients with glottal gap were selected. Of all patients, 14 patients had sulcus vocalis and 12 patients had muscle tension dysphonia (MTD). Voice therapies were performed 1.9 sessions in average. GRBAS, jitter, shimmer, noise to harmonic ratio, semitone range, closed quotient_vowel and maximum phonation time were compared before and after the therapies. In addition, changes of glottal gap and MTD severity were evaluated. Results Statistically significant improvement was observed. MTD improvement was observed only among the patients with glottal gap improvement. Also sulcus vocalis group showed the statistically significant improvement. Conclusion 'Gliding and humming' was effective to the patients with glottic gap and sulcus vocalis. Also, among patients who have both glottic gap and MTD, the data suggests that voice therapy for glottic gap also makes improvement in MTD.
성대폴립, 성대낭종 환자들의 Laryngeal Micro Surgery 수술 전, 후 공기역학적 비교: Running Speech 중심으로
문태훈,심미란,황연신,김근전,이동현,선동일 대한후두음성언어의학회 2019 대한후두음성언어의학회지 Vol.30 No.2
Background and Objectives For patients with polyps and cysts, glottal gaps resulting from their lesions have negative respiratory effects when they vocalize. Phonatory Aerodynamic System is clinically used, but is often limited in the measurement of vowels. So the researchers attempted to verify the usefulness of Phonatory Aerodynamic System by comparing differences in respiratory characteristics and patterns which can be measured by the level of connected speech. Materials and Method Among the subjects who were diagnosed through a stroboscopy, there were 33 patients with polyps and 23 patients with cysts. Then, 36 subjects who were found to have no specific findings through a stroboscopy and perceptual test were selected to the normal group. We compared respiratory characteristics and patterns. And compared vocal polyps and cysts before and after laryngeal micro surgery (LMS). Results First, difference in respiratory patterns between the normal group and the patients with polyps and cysts were examined to show that breath groups, breath group syllables, and expiratory · inspiratory volume were significantly higher in the polyp/cyst group than those in the normal group, indicating that precision was lowered during the conversation, due to reduction in speech intelligibility and interruption of communication. Second, there were significant differences in maximum phonation time, mean flow rate, and subglottal pressure among respiratory characteristics, breath groups, breath group syllables, and inspiratory volume before and after LMS, which appeared to be similar to the normal group. Conclusion The understanding of respiratory characteristics and patterns produced by patients in connected speech which is most similar to natural speech was found to be the objective and useful method for examining characteristics of the subjects.
실용음악 전공 학생들의 발성치료 전후 성대 및 음성 특성 비교
황연신,심미란,김철준,최용석,김상연,최찬호,선동일,Hwang, Yeon Shin,Shim, Mi Ran,Kim, Chul Jun,Choi, Yong Suk,Kim, Sang Yeon,Choi, Chan Ho,Sun, Dong Il 대한후두음성언어의학회 2016 대한후두음성언어의학회지 Vol.27 No.2
Background and Objectives : Many students major in applied music singing have various difficulties in phonation when they sing or perform. But the studies about this cause are lack. The purpose of this study is whether singing voice therapy based on vocal music is effective or not to applied music singing students that have difficulties in phonation. Materials and Methods : Singing voice therapy program had been done to 8 students major in applied music singing during 7 weeks. We did laryngeal stroboscope, perceptual, acoustic and aerodynamic examination before and after singing voice therapy to this students. And we studied post examination results have differences or improvements compared to pre examination results. Results : Four male students don't have effective improvement after singing voice therapy but they shows slight improvement in aerodynamic examination values compared to pre examination values. Four female students shows improvement in aerodynamic examination values and pitch range increasing. Above all, one student's vocal nodules disappeared after singing voice therapy. Conclusion : Singing voice therapy based on music singing is very effective to the students major in applied music singing. Above all, their curriculum in applied music singing should include phonation training based on music singing before training perform and technic practices.