• Archives of Pharmacal Research
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• 제16권4호
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• pp.271-275
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• 1993

In order to prepare a novel human insulin analogue suhbstituted with homoserine at B$^{30}$ / position, (B$^{30}$ /-homoserine) human insulin, a synthetic gene was designed by linking directly a gene for B chain with that for A chain. This gene was constructed by enzymatic joining of 10 different synthetic oligonucleotides, and then inserted at the polylinker region of pUC19 plasmid. To achieve a high level of gene expression, the gene fusion technique region of pUC19 plasmid. To achieve a high level of gene expression, the gene fusion technique was employed using amino terminal regions of lacZ gene up to Clal or hpal, and either of them has been located under tac promoter. The chemical induction of these fused genes by isopropyl-.betha.-D-thiogalactopyranoside (IPTG) gave a satisfactory level of expression in Escherichia coli harboring the ocnstructed plasmids. It was observed that the fused gene product as a single chain insulin precusor was produced more than 30% of total cell protein of E. coli as a form of inclusion body.

• Biotechnology and Bioprocess Engineering:BBE
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• 제1권1호
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• pp.9-12
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• 1996

In order to produce the single chain precursor of a novel human insulin analogue, (B30-Homoserine) insulin, the fermentative behaviors of Escherichia coli JM103 were studied, which harbors pKBA plasmid carrying a hybrid gene in which the gene for a single chain precursor was fused with lacZ gene under tac promoter. The maximal induction of gene expression was achieved when more than 0.05 mM of isopropyl-$\beta$-D-thiogalactopyranoside(IPTG) was supplemented to fermentation medium after 4 h cultivation of E. coli, and followed by longer than 2-h fermentation. The hybrid protein of the single chain insulin precursor was isolated from cytoplasmic inclusion bodies by dissolving in 8M urea solution, and purified through DEAE-Sephacel and Sephadex G-200 column chromatographies with a recovery of 35%. The finally purified hybrid protein showed a single band on sodium dodecyl sulfate-polyacrylamide gel.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XII)
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• pp.34-38
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• 2003

For the production of $B^{30}-homoserine$ insulin analog as a novel anti-diabetic drug, the fermentative study was attempted for the maximal gene expression of HTS-fused $B^{30}-homoserine$ insulin precursor in the recombinant Escherichia coli cells. In a batch fermentation, the maximal production of insulin precursor as much as 38.95 mg/L-h, which occupied more than 12.8% of total cell protein. was achieved when the gene expression was induced by 0.5 mM IPTG at the middle logarithmic growth phase. The HTS-fused $B^{30}-homoserine$ insulin precursor was recovered from a batch culture through the processes of cell harvest, collection of insoluble fraction after sonication and purification by nickel affinity column chromatography. The isolated insulin precursor was 14 mg/L with a recovery yield of 35.9% of expressed gene product. The insulin A and B chain mixture was recovered after the insulin precursor was subjected to CNBr cleavage and purified by nickel affinity column chromatography. The isolated insulin chains were then sulfitolyzed with sodium thiosulfat and sodium tetrathionate, and reconstituted to insulin analog with ${\beta}-mercaptoethanol$, followed by purification with CM-Sepharose C-25 column chromatography.

• BMB Reports
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• 제33권2호
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• pp.120-125
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• 2000

M2PI is an active single chain mini-proinsulin with a 9-residue linker containing the turn-forming sequence 'YPGDV' between the B- and A-chains, but which retains about 50% of native insulin receptor binding activity. The refolding efficiency of M2PI is higher than proinsulin by 20-40% at alkaline pH, and native insulin is generated by the enzymatic conversion of M2PI. The solution structure of M2PI was determined by NMR spectroscopy. The global structure of M2PI is similar to that of native insulin, but the flexible linker between the B- and A-chains perturbed the N-terminal A-chain and C-terminal B-chain. The helix in the N-terminal A-chain is partly perturbed and the ${\beta}$-turn in the B-chain is disrupted in M2PI. However, the linker between the two chains was completely disordered indicating that the designed turn was not formed under the experimental conditions (20% acetic acid). Considering the fact that an insulin analogue, directly cross-linked between the C-terminus of the B-chain and the N-terminus of the A-chain, has negligible binding activity, a flexible linker between the two chains is sufficient to keep binding activity of M2PI, but the perturbed secondary structures are detrimental to receptor binding.

• 한국동물학회지
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• 제38권3호
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• pp.426-432
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• 1995

유선 조직에서 베타-락토글로불린 유전자의 발현은 프롤락틴, 글루코코르티코이드 및 인슐린등의 유촉진 호르몬들에 의해서 강력하게 유도된다. 이와 같은 호르몬 유도의 조절 기작을 규명하기 위하여, 배양 유선 세포인 HC11 세포에서 염소 베타-락토글로불린 유전자 프로모터의 유촉진 호르몬에 대한 반응을 분석하였다. 베타-락토글로불린 프로모터의 5'- 조절 부위를 연쇄적으로 제거한 발현 실험에서 호르몬 유도를 크게 변화 시키는 두 지역이 관찰되었다. 조절 부위의 -1692의 상류지역은 하류 프로모터를 강력하게 활성화 시키는 부위로, 주로 글루코코르티코이드 유도체인 덱사메타손의 작용을 매개하였다. 그러나 두번째 지역의 유도 작용은 인슐린 처리를 병행하지 않을 경우 상류 조절부위에 의해 억제되었다. 이러한 결과는, 유선세포에서 유촉진 호르몬들에 의한 베타-락토글로불린 프로모터 활성 유도가 인슐린에 의한 탈 억제화와 글루코코르티코이드 및 프롤락틴에 의한 활성화의 복합 조절에 의해서 이루어질 것이라는 점을 시사한다. 두번째 지역에 의한 덱사메타손 유도는 -700 부근의 글루코코르티코이드 수용체 결합 부위에 의해서 매개되는 것으로 추정된다.

• The Korean Journal of Physiology and Pharmacology
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• 제18권3호
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• pp.255-261
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• 2014

Essential fatty acid (EFA) is known to be required for the body to function normally and healthily. However, the effect of EFA on glucose uptake in skeletal muscle has not yet been fully investigated. In this study, we examined the effect of two EFAs, linoleic acid (LA) and ${\alpha}$-linolenic acid (ALA), on glucose uptake of C2C12 skeletal muscle cells and investigated the mechanism underlying the stimulatory effect of polyunsaturated EFAs in comparison with monounsaturated oleic acid (OA). In palmitic acid (PA)-induced insulin resistant cells, the co-treatment of EFAs and OA with PA almost restored the PA-induced decrease in the basal and insulin-stimulated 2-NBDG (fluorescent D-glucose analogue) uptake, respectively. Two EFAs and OA significantly protected PA-induced suppression of insulin signaling, respectively, which was confirmed by the increased levels of Akt phosphorylation and serine/threonine kinases ($PKC{\theta}$ and JNK) dephosphorylation in the western blot analysis. In PA-untreated, control cells, the treatment of $500{\mu}M$ EFA significantly stimulated 2-NBDG uptake, whereas OA did not. Phosphorylation of AMP-activated protein kinase (AMPK) and one of its downstream molecules, acetyl-CoA carboxylase (ACC) was markedly induced by EFA, but not OA. In addition, EFA-stimulated 2-NBDG uptake was significantly inhibited by the pre-treatment of a specific AMPK inhibitor, adenine 9-${\beta}$-D-arabinofuranoside (araA). These data suggest that the restoration of suppressed insulin signaling at PA-induced insulin resistant condition and AMPK activation are involved at least in the stimulatory effect of EFA on glucose uptake in C2C12 skeletal muscle cells.

• Clinical and Experimental Pediatrics
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• 제50권6호
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• pp.565-569
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• 2007

목 적 : 란투스는 최대효과 없이, 24시간 동안 효과가 지속되기 때문에 NPH에 비해 생리적인 기저 인슐린 대체 제제이다. 1형 당뇨병 소아 및 청소년에서 란투스와 휴마로그의 병합 치료와 혼합형 인슐린의 혈당 조절 효과를 비교하기 위해 본 연구를 시행 하였다. 방 법 : 혼합형 인슐린에서 란투스와 휴마로그의 병합 치료로 인슐린 요법을 바꾼 1형 당뇨병을 가진 20세 미만의 환아 25명을 대상으로 란투스 시작 전과 시작 6개월 후의 하루 인슐린 투여량, 월간 저혈당 횟수, 공복시 혈당, C-peptide 농도 및 당화혈색소를 비교하였다. 이들 중 11명의 환아와 혼합형 인슐린을 사용하는 10명의 환아를 대상으로 24시간 자가 혈당 검사를 시행하여 매 식사 30분 전과 식후 30분 간격으로 2시간 동안의 혈당과 취침 전의 혈당 비교하였다. 결 과 : 란투스 치료 6개월 후 저혈당 빈도가 월간 15.1회에서 7.6회로 50% 감소하였으며, 특히 야간 저혈당 빈도는 월간 6.7회에서 2.5회로 67% 감소하였다. 당화혈색소는 란투스 치료 6개월 후 9.3%에서 8.7%로 감소하였다. 24시간 혈당 검사에서는 란투스를 사용하는 군에서 아침 식후 30분, 60분, 90분, 120분에서의 혈당이 혼합형 인슐린을 사용하는 군보다 유의하게 낮았고, 24시간 평균 혈당은 란투스 사용군이 $164.1{\pm}78.2mg/dL$로 혼합형 인슐린 사용군의 $211.5{\pm}108.7mg/dL$보다 유의하게 낮았다. 결 론 : 1형 당뇨병 소아 및 청소년에서 란투스와 휴마로그의 병합 치료는 혼합형 인슐린과 비교하여 혈당 조절에 보다 효과적이고, 특히 야간 저혈당 감소에 유효한 것으로 생각된다.

• Clinics in Orthopedic Surgery
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• 제10권4호
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• pp.455-461
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• 2018

Background: Surgical-site, multimodal drug injection has recently evolved to be a safe and useful method for multimodal pain management even in patients with musculoskeletal trauma. Methods: Fifty consecutive patients who underwent plating for mid-shaft and distal clavicular fractures were included in the study. To evaluate whether surgical-site injections (SIs) have pain management benefits, the patients were divided into two groups (SI and no-SI groups). The injection was administered between the deep and superficial tissues prior to wound closure. The mixture of anesthetics consisted of epinephrine hydrochloride (HCL), morphine sulfate, ropivacaine HCL, and normal saline. The visual analogue scale (VAS) pain scores were measured at 6-hour intervals until postoperative hour (POH) 72; stress biomarkers (dehydroepiandrosterone sulfate [DHEA-S], insulin, and fibrinogen) were measured preoperatively and at POH 24, 48, and 72. In patients who wanted further pain control or had a VAS pain score of 7 points until POH 72, 75 mg of intravenous tramadol was administered, and the intravenous tramadol requirements were also recorded. Other medications were not used for pain management. Results: The SI group showed significantly lower VAS pain scores until POH 24, except for POH 18. Tramadol requirement was significantly lower in the SI group until POH 24, except for POH 12 and 18. The mean DHEA-S level significantly decreased in the no-SI group ($74.2{\pm}47.0{\mu}g/dL$) at POH 72 compared to that in the SI group ($110.1{\pm}87.1{\mu}g/dL$; p = 0.046). There was no significant difference in the insulin and fibrinogen levels between the groups. The correlation values between all the biomarkers and VAS pain scores were not significantly different between the two groups (p > 0.05). Conclusions: After internal fixation of the clavicular fracture, the surgical-site, multimodal drug injection effectively relieved pain on the day of the surgery without any complications. Therefore, we believe that SI is a safe and effective method for pain management after internal fixation of a clavicular fracture.

• 재활간호학회지
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• 제6권2호
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• pp.152-163
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• 2003

Purpose; This study was done to investigate the effect of foot reflexology on vital signs, general fatigue, foot fatigue, mood, and blood glucose levels in noninsulin dependent patients. Method: The Research design of this study was nonequivalent control group quasi-experimental design. 18 patients were assigned to the experimental group, 24 patients to the control group. The data were obtained diaberic patients with ambulatory endocrine outpatients clinic patients from 40 years old to 70 years old. Experimental groups received foot reflex massage for 30minutes three times/week every other days, and Control groups did not received foot reflex massage. The dependent variables were blood pressure, pulse rate, visual analogue scale for general fatigue, foot fatigue, mood, and blood sugar levels. Data were analyzed with $X^2$ test, t-test and repeated measure ANOVA at .0.05 level of significance. Results: There were significant difference in the pulse rate, general fatigue, foot fatigue and mood according to group and time between pre and post foot reflexology. But this research did not prove to decrease blood sugar levels. Conclusions : Foot reflexology can imorove pulse rate, general and foot fatigue, and mood status in diabetus patients. So further research need to explore the effect of decreasing of blood sugar levels.

• Clinical and Experimental Pediatrics
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• 제50권7호
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• pp.678-685
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• 2007

목 적 : 조기사춘기 여아에게 성선자극호르몬 방출호르몬 효능약제(gonadotropin releasing hormone analogue, GnRHa)와 성장호르몬(growth hormone, GH)의 병합치료를 시행하여 성인신장획득에 대한 효과를 알아보고자 하였다. 방 법 : 2001년 1월부터 2004년 1월까지 조기사춘기로 소아과 외래를 내원한 여아 중 예측성인신장이 -2 표준편차점수 이하로 표적키보다 훨씬 작은 23명($9.73{\pm}1.59$세)을 대상으로 초경을 시작하지 않은 군(Group 1, n=19, $9.59{\pm}1.59$세)과 초경을 시작한 군(Group 2, n=4, $10.38{\pm}1.44$세)으로 나누어 GnRHa와 GH의 병합치료를 시행하고, 치료 시작 전후 및 두 군 간의 역연령, 골연령, 성장속도, 신장, 예측성인신장, 혈청 IGF-1, IGFBP-3 치, BMI를 비교 분석하였다. 결 과 : 치료 시작 시의 두 군의 역연령, 성장속도, BMI는 유의한 차이는 없었으나 골연령 및 골연령과 역연령의 차이는 Group 2에서 Group 1보다 유의하게 증가되어 있었다. 신장은 Group 1의 평균이 $130.7{\pm}7.4cm$, Group 2의 평균이 $140.7{\pm}6.0cm$으로 유의하게 차이가 있었으나 표적키, 예측성인신장, 혈청 IGF-1과 IGFBP-3은 유의한 차이가 없었다. 치료 종료 시의 Group 1의 골연령은 평균 $12.18{\pm}1.02$세, Group 2의 골연령은 평균 $13.56{\pm}0.83$세로 치료 시작 전과 마찬가지로 Group 2에서 유의하게 높았으나, 골연령과 역연령의 차이, 신장, 그리고 IGF-1은 차이가 없었다. 전체에서 골연령과 역연령의 차이가 치료 시작 전에 평균 $2.50{\pm}2.46$세, 치료 종료 후에 평균 $1.10{\pm}0.97$세로 감소되었고, 역연령에 대한 골연령의 비도 $1.22{\pm}0.16$에서 $1.10{\pm}0.10$으로 감소되어서 결국 골연령의 증가는 현저히 감소하였다. 성장 속도 역시 평균 $7.46{\pm}2.44cm$/년에서 $5.65{\pm}2.08cm$/년으로 감소하였다. 예측성인신장은 치료 전에 평균 $146.6{\pm}3.50cm$, 치료 후에 평균 $153.8{\pm}4.7cm$으로 현저히 증가하였으며 신장의 표준편차점수도 $-0.66{\pm}1.02$ SDS에서 $-0.34{\pm}1.04$ SDS로 유의하게 증가하였다. BMI는 치료 전 평균 $18.80{\pm}2.35kg/m^2$, 치료 후에 평균 $20.4{\pm}2.53kg/m^2$로 유의하게 증가하였고, 혈청 IGF-1은 치료 후에 유의하게 증가하였으나 IGFBP-3는 유의한 변화가 없었다. 치료 전후에 두 군에서 각 성장 변수의 변화를 보면 골연령과 역연령의 차이는 유의하게 감소하였고 신장, 예측성인신장, BMI는 유의하게 증가하였다. 그러나 혈청 IGF-1의 경우, Group 1에서는 치료 후에 현저히 증가하였으나 Group 2에서는 별 차이가 없었다. 혈청 IGFBP-3는 모두에서 유의한 차이가 없었다. 결 론 : 조기사춘기 여아에게 GnRHa와 GH의 병합치료를 시행했을 때, 골연령의 증가 속도와 성장속도가 감소하고, 혈청 IGF-1이 증가함으로써 예측성인신장이 의미 있게 증가하며 표적키에 가깝게 증가한다. 또한 IGF-1의 변화를 보면 초경을 시작하지 않은 조기사춘기 여아가 초경을 시작한 경우보다 성장획득에 대한 가능성이 더 클 것으로 생각된다.