• The Journal of Korean Physical Therapy
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• 제19권5호
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• pp.65-76
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• 2007

Purpose: It is generally accepted that smooth muscle contraction is triggered by intracellular $Ca^{2+}$ ($[Ca^{2+}]_i$) released from intracellular $Ca^{2+}$ stores such as sarcoplasmic teticulum (SR) and from the extracellular space. The increased $[Ca^{2+}]^i$ can phosphorylate the 20,000 dalton myosin light chain $(MLC_{20})$ by activating MLC kinase (MLCK), and this initiates smooth muscle contraction. In addition to the $[Ca^{2+}]_i$MACK-tension pathway, a number of intracellular signal molecules, including mitogen-activated protein kinase (MAPK), protein kinase C (PKC) and others, play important roles in the regulation of smooth muscle contraction. However, the mechanisms regulating contraction of depletion of SR $Ca^{2+}$ in mouse gastric smooth muscle strips is not still clear. Methods: To investigate the rotes of $Ca^{2+}$ influx and SR $Ca^{2+}$ release channel on gastric motility, isometric contraction and $[Ca^{2+}]_i$ were examined in mouse gastric smooth muscle strips. Results: High KCl, ryanodine, an activator of $Ca^{2+-}$induced $Ca^{2+}$ release channel, and cyclopiazonic acid (CPA), an inhibitor of SR $Ca^{2+-}$ATPase evoked a sustained increase in muscle contraction and $[Ca^{2+}]_i$. These increases induced by high KCl, ryanodine, and CPA were partially blocked by application of verapamil ($10{\mu}M$), a L-type $Ca^{2+}$ channel inhibitor. Additionally, in $Ca^{2+-}$free solution (1 mM EGTA), ryanodine and CPA had no effect contraction and $[Ca^{2+}]_i$ in fundic muscle strips. Conclusion: These results that extracellular $Ca^{2+}$ influx and depletion of SR trigger $Ca^{2+}$ influx through verapamil-sensitive $Ca^{2+}$ channel, and extracellular and SR $Ca^{2+}$ store may functionally involve in the subcellular $Ca^{2+}$ mobilization in mouse gastric muscle.

• Asian Pacific Journal of Cancer Prevention
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• 제14권11호
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• pp.6557-6562
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• 2013

Objective: Specific promoters could improve efficiency and ensure the safety of gene therapy. The aim of our study was to screen examples for lung cancer. Methods: The firefly luciferase gene was used as a reporter, and promoters based on serum markers of lung cancer were cloned. The activity and specificity of seven promoters, comprising CEACAM5 (carcinoembryonic antigen, CEA), GRP (Gastrin-Releasing Peptide), KRT19 (cytokeratin 19, KRT), SFTPB (surfactant protein B, SP-B), SERPINB3 (Squamous Cell Carcinoma Antigen, SCCA), SELP (Selectin P, Granule Membrane Protein 140kDa, Antigen CD62, GMP) and DKK1 (Dickkopf-1) promoters were compared in lung cancer cells to obtain cancer-specific examples with strong activity. Results: The CEACAM5, DKK1, GRP, SELP, KRT19, SERPINB3 and SFTPB promoters were cloned. Furthermore, we successfully constructed recombinant vector pGL-CEACAM5 (DKK1, GRP, SELP, KRT19, SERPINB3 and SFTPB) contained the target gene. After cells were transfectedwith recombinant plasmids, we found that the order of promoter activity from high to low was SERPINB3, DKK1, SFTPB, KRT19, CEACAM5, SELP and GRP and the order for promoters regarding specificity and high potential were SERPINB3, DKK1, SELP, SFTPB, CEACAM5, KRT19 and GRP. Conclusion: The approach adopted is feasible to screen for new tumour specific promoters with biomarkers. In addition, the screened lung-specific promoters might have potential for use in lung cancer targeted gene therapy research.

• 한방안이비인후피부과학회지
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• 제29권1호
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• pp.47-64
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• 2016

Objective : This study was carried out to investigate the effect of Hwangryeonhaedoktang pharmacopuncture solution (HRHDT) and microneedle therapy system (MTS) on hair growth in an alopecia model of C57BL/6N mice.Methods : Six-week old mice were depilated and separated in 4 groups ; CON (saline), MXD (3% Minoxidil), MTS and HRHDT+MTS. The treatments were applied twice a week for 17 days. The hair growth was determined photographically. The hair density, thickness and length were identified by Folliscope and the weights of body and organs were measured. In dorsal skin tissue, the expression of hair growth-related gene and protein was analyzed by RT-PCR. In addition, the hair follicles in the dermis were observed by H&E staining.Results : The promotion of hair growth was observed in HRHDT+MTS and MTS compared to CON. The hair density, thickness and length were also improved in HRHDT+MTS and MTS compared to CON. The mRNA expression of IGF-1, PRL and PL and the protein expression of VEGF and IGF-1 were increased in HRHDT+MTS and MTS compared to CON. The hair follicles and hair root growth were improved in HRHDT+MTS and MTS compared to CON. In the above results, HRHDT+MTS were more effective than MTS.Conclusions : These results suggest that HRHDT and MTS have a hair growth activity and can be useful for the treatment of alopecia.

• 대한한방소아과학회지
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• 제28권2호
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• pp.72-87
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• 2014

Objectives This study was carried out to investigate the effect of Ganoderma lucidum extract (GLE) and microneedle therapy system (MTS) on hair growth in an alopecia model of C57BL/6N mice. Methods Five-week old mice were depilated and separated in 4 groups ; CON (50% EtOH), MXD (5% Minoxidil), MTS and GLE + MTS. The treatments were applied twice a week for 3 weeks. The hair growth was determined photographically, the hair density, thickness and length were identified by Folliscope and the weights of body and organs were measured. In dorsal skin tissue, the expression of hair growth-related gene and protein was analyzed by RT - PCR or Western blot. In addition, the hair follicles in the dermis were observed by H&E staining. Results The promotion of hair growth was observed in GLE + MTS and MTS compared to CON. The hair density, thickness and length were also improved in GLE + MTS and MTS compared to CON. The mRNA expression of TGF-${\alpha}$, TGF-${\beta}1$, IGF-1, PRL and PL and the protein expression of VEGF and IGF-1 were increased in GLE + MTS and MTS compared to CON. The hair follicles and hair root growth were improved in GLE + MTS and MTS compared to CON. In the above results, GLE + MTS were more effective than MTS. Conclusions These results suggest that GLE and MTS has a hair growth activity and can be useful for the treatment of alopecia.

• BMB Reports
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• 제43권8호
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• pp.561-566
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• 2010

Though protein transduction domains (PTDs) are well known for the delivery of exogenous therapeutic proteins into living cells, the overall low efficiency of transduction is a serious obstacle. We investigated the effect of bog blueberry anthocyanins (BBA) on protein transduction efficiency and found that BBA enhanced the transduction efficiencies of Tat-SOD fusion protein into HeLa cells and mice skin. The enzymatic activities in the cells and skin tissue in the presence of BBA were markedly increased compared to controls. Further, BBA did not demonstrate any cell toxicity at various concentrations. Although the mechanism is not fully understood, we suggest that BBA might alter the conformation of the membrane, which would indicate that BBA can be used as a protein transduction enhancer for the efficient delivery of therapeutic proteins for a variety of disorders.

• BMB Reports
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• 제51권7호
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• pp.362-367
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• 2018

A major feature of type 1 diabetes mellitus (T1DM) is hyperglycemia and dysfunction of pancreatic ${\beta}$-cells. In a previous study, we have shown that Tat-DJ-1 protein inhibits pancreatic RINm5F ${\beta}$-cell death caused by oxidative stress. In this study, we examined effects of Tat-DJ-1 protein on streptozotocin (STZ)-induced diabetic mice. Wild type (WT) Tat-DJ-1 protein transduced into pancreas where it markedly inhibited pancreatic ${\beta}$-cell destruction and regulated levels of serum parameters including insulin, alkaline phosphatase (ALP), and free fatty acid (FFA) secretion. In addition, transduced WT Tat-DJ-1 protein significantly inhibited the activation of $NF-{\kappa}B$ and MAPK (ERK and p38) expression as well as expression of COX-2 and iNOS in STZ exposed pancreas. In contrast, treatment with C106A mutant Tat-DJ-1 protein showed no protective effects. Collectively, our results indicate that WT Tat-DJ-1 protein can significantly ameliorate pancreatic tissues in STZ-induced diabetes in mice.

• Journal of Nutrition and Health
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• 제48권3호
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• pp.211-220
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• 2015

본 연구는 지속적 신대체요법을 받은 급성 신손상 환자 239명을 대상으로 영양공급 충족률이 환자의 임상적인 결과에 미치는 영향을 알아보고자 수행하였으며 대상자의 영양요구량 및 실제공급량을 조사하여 영양공급 충족률에 따른 임상결과를 비교하였다. 대상자의 평균 에너지 충족률이 68.06%, 평균 단백질 충족률이 43.13%로 나타났고 체중 당 에너지 공급량은 16.62 kcal/kg/일, 단백질 공급량은 0.63 g/kg/일로 지침에서 권장하는 요구량에 미치지 못하는 수준이었다. 에너지 충족률 70%, 단백질 충족률 50% 기준으로 두 군간 임상결과의 차이를 관찰한 결과 에너지 충족률이 높은 군에서 총 재원일수, 중환자실 재원일수와 지속적 신대체요법 적용일수, APACHE II score가 유의하게 감소하는 것으로 분석되었고 단백질 충족률이 높은 군에서는 총 재원일수, 중환자실 재원일수, 지속적 신대체요법 적용일수, APACHE II score 이외에 사망률도 유의하게 감소한 것으로 분석되었다. 생화학적 검사 결과의 경우 에너지 충족률 70% 이상 충족군에서는 헤마토크릿 수치가, 단백질 충족률 50% 이상 충족군에서는 총 임파구수, 헤마토크릿 수치가 유의하게 개선된 것으로 나타났다. 사망환자를 제외하고 영양공급량과 임상결과를 나타내는 항목의 상관관계를 분석한 결과 에너지 공급량과 중환자실 재원일수가 의미 있는 음의 상관관계를 가지는 것으로 나타났으며 영양공급량과 생화학적 검사 결과의 상관관계를 파악한 결과 에너지 공급량에 따라서는 유의한 상관계수가 도출되지 않았고 단백질 공급량과는 따라서는 헤마토크릿과 칼슘이 유의한 양의 상관관계를 가지는 것으로 나타났다. 본 연구 결과 영양공급 충족률이 높은 군에서 임상적인 결과 및 영양상태와 관련한 생화학적 검사결과가 개선되는 것으로 나타나 요구량에 적정한 영양공급이 환자의 치료에 긍정적인 영향을 준다는 것을 확인할 수 있었다. 이를 위해서 의료진의 타당한 영양처방이 중요하겠으며 영양지원의 체계적인 관리를 위해 다학제적인 접근과 함께 영양공급의 적정성을 지속적으로 모니터링하는 것이 중요하겠다.

• IMMUNE NETWORK
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• 제4권1호
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• pp.7-15
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• 2004

Background: The heat shock proteins (HSPs) play an important role in cellular protection mechanisms against physical or chemical stresses. In this study scFv antibodies specific for human HSP70.1 were isolated from a semi-synthetic human scFv library with the ultimate goal of developing anti-HSP70.1 intracellular antibody (intrabody) that may offer an attractive alternative to gene targeting to study the function of the protein in cells. Methods: A semi-synthetic human scFv display library ($5{\times}10^{8}$ size) was constructed using pCANTAB-5E vector and the selection of the library against bacterially expressed recombinant human HSP70.1 was attempted by panning. Results: Three positive clones specific for recombinant HSP70.1 were identified. All three clones used $V_{H}$ subgroup III. On the other hand, $V_{L}$ of two clones belonged to the kappa light chain subgroup I, but the other utilized $V_{k}$ subgroup IV Interestingly, these scFv molecules specifically reacted to the recombinant HSP70.1, yet failed to recognize native HSP70 induced in U937 human monocytic cells by heat treatment. Conclusion: Our results indicated that affinity selection of an scFv phage display library using recombinant antigens produced in E. coli might not guarantee the isolation of scFv antibody molecules specific for a native form of the antigen. Therefore, the source of target antigens needs to be chosen carefully in order to isolate biofunctional antibody molecules.

• Asian Pacific Journal of Cancer Prevention
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• 제16권6호
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• pp.2425-2430
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• 2015

Clinical resistance to chemotherapeutic agents is one of the major hindrances in the treatment of human cancers. EHZ2 is involved in drug resistance and is overexpressed in drug-resistant cancer cell lines. In this study, we investigated the effects of EHZ2 on cisplatin -resistance in A549/DDP and AGS/DDP cells. EHZ2 mRNA and protein were found to be significantly overexpressed in A549/DDP and AGS/DDP cells, compared to parental cells. EHZ2 siRNA successfully silenced EHZ2 mRNA and protein expression. Proliferation was inhibited and drug resistance to cisplatin was improved. Flow cytometry showed that silencing of EHZ2 arrested A549/DDP and AGS/DDP cells in the G0/G1 phase, increasing apoptosis, rh-123 fluorescence intensity and caspase-3/8 activities. Silencing of EHZ2 also significantly reduced the mRNA and protein expression levels of cyclin D1 and MDR1,while up-regulating p15, p21, p27 and miR-218 in A549/DPP cells. Furthermore, silencing of EHZ2 also significantly increased the expression level of tumor suppressor factor miR-218. We also found down-regulating EHZ2 expression increased methylation in A549/DDP and AGS/DDP cells. This study demonstrates that drug resistance can be effectively reversed in human cisplatin-resistant lung and gastric cancer cells through delivery of siRNAs targeting EHZ2.

• 국제물리치료학회지
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• 제6권2호
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• pp.846-851
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• 2015

This neurological damage accelerates the infection reaction of cells and apoptosis at the time of reperfusion after ischemia occurs. BCL-2/BCL-2 allogeneic begeminum has a function of suppressing the apoptosis of cells, and thus it is inferred that the susceptibility of cells to apoptosis is determined by the amount of allogeneic begeminum present which is determined based on the amount of BAX. Ischemia was induced in SD mice by occluding the common carotid artery for 5 minutes, after which blood was re-perfused. NEES was applied to acupuncture points, at 12, 24, and 48 hours post-ischemia on the joksamri, Hapgok. Protein expression was investigated through BAX antibody immuno-reactive cells in the cerebral nerve cells and Western blotting. The results were as follows: In the present study as well, as a result of observation of the change in the number of the BAX reaction cells after the inducement of GI, there was the aspect of most of the BAX reaction cells being observed in the corpus striatum area of the GI group 24 hours after the inducement of ischemia. This revealed the same results as those of previous studies in which the change in the number of BAX reaction cells occurred in all areas while ischemia was in progress. The change in the expression of BAX protein after 24 hours showed that there was a very significant reduction in the NEES group compared to the GI group (p<.01). As a result, a greatest amount of change in the number of BAX immunoreactive cells related to apoptosis 24 hours after ischemia appeared in the NEES group. This study that ischemia increases the expression of BAX that induces apoptosis. Thus, it is determined that ischemia is the main cause of the apoptosis of neurons, and this study reveals that low frequency needle electrode electrical stimulation has the effect of blocking the apoptosis of neurons by reducing protein related to the apoptosis of cells that has increased after ischemia has occurred.