• 한국잠사곤충학회지
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• 제35권2호
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• pp.129-133
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• 1993

Bacillus thruingiensis var. kurstaki HD-1의 내독소 단백질 유전자의 발현기작을 규명하기 위하여 이 균으로부터 내독소 단백질 유전자가 존재하는 것으로 확인된 29Md와 44Md plasmid를 분리한 후, Sau3AI 제어효소로 부분절단하고, pBR322 BamHI site에 ligation하여, E. coli HB101 strain에 transformation시켜, 3,000여개의 Ampr/Ters한 colony를 얻어 면역학적 방법과 살충적 검정으로 통해 재조합 균주 KC1을 얻었다. PKC1 plasmid DNA는 vector DNA를 포함하여 약 12kb 정도의 크기를 가지며, B. t k HD-1 내독소 단백질과 이동도가 같거나 일치하는 132kd, 117kd의 KC1 specific band 2개를 얻었다. KC1 cell extract를 첨식한 결과 약 80% 치사율을 보였다.

• BMB Reports
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• 제31권1호
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• pp.83-88
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• 1998

Recently a B cell surface molecule, CD40, has emerged as a receptor mediating a co-stimulatory signal for B cell proliferation and differentiation. To investigate the mechanism of synergy between interleukin-4 (IL-4) and CD40 ligation in B cell activation, we have examined the effect of CE40 cross-linking on the IL-4 receptor expression in human B cells using anti-CE40 antibody. We observed that IL-4 and anti-CD40 both induce IL-4 receptor gene expression with a rapid kinetics resulting in a noticeable accumulation of IL-4 receptor mRNA within 4 h. While IL-4 caused a dose-dependent induction of surface IL-4 receptor expression, the inclusion of anti-CD40 in the IL-4-treated culture, further up-regulated the IL-4-induced IL-4 receptor expression as analyzed by flow cytometry. Pretreatment of B cells with inhibitors of protein tyrosine kinase (PTK) resulted in a significant inhibition of both the IL-4- and anti-CD40-induced IL-4 receptor mRNA levels, while protein kinase C (PKC) inhibitors had no effects. These results suggest that IL-4 and CD40 ligation generate B cell signals, which via PTK-dependent pathways, lead to the synergistic induction of IL-4 receptor gene expression. The rapid induction of IL-4 receptor gene expression through the tyrosine kinase-mediated signal transduction by B cell activating stimuli, would provide cells capacity for an efficient response to IL-4 in the early phase of IL-4 action, and may in part constitute the molecular basis of the reported anti-CD40 co-stimulatory effect on the IL-4-induced response.

• 대한한의학회지
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• 제26권3호
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• pp.13-26
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• 2005

Objectives : Applying herbal-acupuncture using Ursi Fet into Zusanli (UZ) on to the atopic dermatitis (AD) in mice to study changes in external dermal formation, change of leukocytes in vasculature, change of lipid formation in stratum corneum and distribution of ceramide. This study was done through forcing an injury to the mice's back skin which damages the lipid protection formation in the stratum corneum. Methods : The AD which was caused intentionally using the external application on the mice's back skin was treated with VB; the change of leukocytes in the vasculature was identified through optima 5.2 and Student's t-test and the results were made into a dermal formation graph. Results : After dispensing UZ into the AD, the dermal injury decreased. The recovery of the lipid protection formation which includes lipid and ceramide in the stratum comeum (for suppressing acute inflammation due to factors such as PKC, $TNF-\alpha,\;IL-1\beta$, which controlled the secretion of the relating inflammatory cytokine) also went on to show a decrease of both angiogenesis and degranulated mast cells. In addition, the decrease of epithelial injury also caused the growth of cells to decrease in the stratum basale and cytoclasis. In the vasculature, the leukocytes were also decreased na this could relate to a decrease in AD. Conclusions : UZ has an effect on AD by suppressing dermal injury through the recovery of the lipid protection formation in the stratum corneum.

• 대한한의학회지
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• 제25권3호
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• pp.137-148
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• 2004

Objectives : To study change of external dermal formation, change of leukocytes in vasculature, change of lipid formation in stratum corneum and distribution of ceramide through administering Sihocheonggan-san (SC) extract on to the control (CON). This study was done through forcing injury to mice's back skin which have lipid protection formation in stratum corneum. Materials and Methods : The CON to which damage was caused intentionally using the external application on the mice's back skin used the SC. The change of leukocytes in vasculature were identified through optima 5.2 and Student's t-test and the results were made into a dermal formation graph. Results : After dispensing SC extract into the CON, dermal injury was decreased. Especially, recover of lipid protection formation, which includes lipid and ceramide in stratum corneum suppressing acute inflammation that some factors are PKC, TNF-α, IL-12B which controlled the secretion of relating inflammatory cytokine, also went onto decrease of angiogenesis, and a decrease of degranulated mast cells was noted. In addition, the decrease of epithelial injury also caused the growth of cells to decrease in stratum basale and cytoclasis. In the vasculature, the leukocytes were also decreased and it could relate to decrease in CON. Conclusions : SC has effect on CON suppressing the dermal injury through recovering of lipid protection formation in stratum corneum.

• 대한물리치료과학회지
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• 제11권1호
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• pp.20-27
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• 2004

It is widely accepted that smooth muscle contraction is triggered by intracellular $Ca^{2+}$ ($[Ca^{2+}]_i$) released from intracellular $Ca^{2+}$ stores such as sarcoplasmic reticulum (SR) and from the extracellular space, The increased $[Ca^{2+}]_i$ can phosphorylate the 20-kDa myosin light chain ($MLC_{20}$) by activating MLC kinase (MLCK), and this initiates smooth muscle contraction. In addition to the $[Ca^{2+}]_i$-MLCK-tension pathway, a number of intracellular signal molecules, including mitogen-activated protein kinase (MAPK), protein kinase C (PKC), phosphatidylinositol 3-kinase (PI3K), and Rho-associated coiled coil-forming protein kinase (ROCK), play important roles in the regulation of smooth muscle contraction. However, the mechanisms regulating contraction of caldesmon (CaD), actin-binding protein, are not entirely elucidated in the presence of $Ca^{2+}$. It is known that CaD tightly interacts with actin and inhibits actomyosin ATPase activity. Therefore, the purpose of the present study was to investigate the roles of $Ca^{2+}$-dependent CaD in smooth muscle contraction. Endothelin-1 (ET-1), G-protein coupled receptor agonist and vasoconstrictor, increased both vascular smooth contraction and phosphorylation of CaD in the presence of $Ca^{2+}$. These results suggest that ET-1 induces contraction and phosphorylation of CaD in rat aortic smooth muscle, which may he mediated by the increase of $[Ca^{2+}]_i$.

• 한국환경독성학회:학술대회논문집
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• 한국환경독성학회 2003년도 추계국제학술대회
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• pp.13-13
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• 2003

Methylmercury (MeHg) is a ubiquitous environmental toxicant that can be exposed to humans by ingestion of contaminated food including fish and bread. MeHg has been suggested to exert its toxicity through its high reactivity to thiols, generation of arachidonic acid and reactive oxygen species (ROS), and elevation of intracellular $Ca^{2+}$ levels (［$Ca^{2+}$］$_{i}$). However, the precise mechanism has not been fully defined. Here we show that phosphatidylcholine-specific phospholipase C (PC-PLC) is a critical pathway for MeHg-induced toxicity. MeHg activated the acidic form of sphingomyelinase (A-SMase) and group IV cytosolic phospholipase $A_2$ ($cPLA_2$) downstream of PC-PLC, but these enzymes as well as protein kinase C were not linked to MeHg's toxicity. Furthermore, MeHg produced ROS, which did not cause the toxicity. However, D6O9, an inhibitor of PC-PLC, significantly reversed the toxicity in a time- and dose-dependent manner in MDCK and SH-5YSY cells. Addition of EGTA to culture media resulted in partial decrease of [$Ca^{2+}$］$_{i}$ and partially blocked cell death. In contrast, D609 completely prevented cell death with parallel decreases in diacylglycerol and ［$Ca^{2+}$］$_{i}$. Together, our findings indicated that MeHg-induced toxicity was caused by elevation of ［$Ca^{2+}$]$_{i}$ through activation of PC-PLC. The toxicity was not attributable to the signaling pathways such as $cPLA_2$, A-SMase, and PKC, or to the generation of ROS.

• The Korean Journal of Physiology and Pharmacology
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• 제4권2호
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• pp.113-120
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• 2000

To investigate the mechanism of smooth muscle contraction induced by emptying of intracellular $Ca^{2+}$ stores, we measured isometric contraction and $^{45}Ca^{2+}$ influx. $CaCl_2$ increased $Ca^{2+}$ store emptying- induced contraction in dose-dependent manner, but phospholipase C activity was not affected by the $Ca^{2+}$ store emptying-induced contraction. The contraction was inhibited by voltage-dependent $Ca^{2+}$ channel antagonists dose dependently, but not by TMB-8 (intracellular $Ca^{2+}$ release blocker). Both PKC inhibitors (H-7 and staurosporine) and tyrosine kinase inhibitors (genistein and methyl 2,5-dihydroxycinnamic acid) significantly inhibited the contraction, but calmodulin antagonists (W-7 and trifluoperazine) had no inhibitory effect on the contraction. The combined inhibitory effects of protein kinase inhibitors, H-7 and genistein, together with verapamil were greater than that of each one alone. In $Ca^{2+}$ store-emptied condition, $^{45}Ca^{2+}$ influx was significantly inhibited by verapamil, H-7 or genistein but not by trifluoperazine. However combined inhibitory effects of protein kinase inhibitors, H-7 and genistein, together with verapamil were not observed. Therefore, this kinase pathway may modulate the sensitivity of contractile protein. These results suggest that contraction induced by emptying of intracellular $Ca^{2+}$ stores was mediated by influx of extracellular $Ca^{2+}$ through voltage-dependent $Ca^{2+}$ channel, also protein kinase C and/or tyrosine kinase pathway modulates the $Ca^{2+}$ sensitivity of contractile protein.

• 대한예방한의학회지
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• 제8권2호
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• pp.13-30
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• 2004

This study was performed to evaluate the effect of Kangwhal-Sokdan tang(KS) on osteoblast function and gene expression. The osteoblast separated from the murine calvariae and MG-63 cell were cultivated to evaluate the cell function and gene expression. The results were summarized as followes. 1) KS increased cell proliferation of murine calvarial cell. 2) KS increased protein synthesis, collagen synthesis and ALP activity of murine calvarial cell. 3) KS increased the survival rate of murine calvarial cell. 4) KS increased the expression of calcitonin receptor and PTH receptor. 5) KS increased the expression of PKA and PKC. 6) KS decreased the expression of $PLA_2$, COX, $PGE_2$ synthase, but increased prostacyclin synthase. 7) KS increased the expression of collagen(type IV) gene. It is concluded that KS might improve the osteoporosis resulted from augumentation of osteoblast proliferation and gene expression.

• The Journal of Korean Physical Therapy
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• 제21권2호
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• pp.109-116
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• 2009

Purpose: $17\beta$-estradiol is the most active endogenous estrogen, which is related to favorable changes in the plasma lipid profile, to relaxation of the coronary vessels, and to a decrease in platelet aggregation and vascular smooth muscle cell migration. However, although the beneficial effect of estrogens on plasma lipoproteins (ie, lowering low-density lipoprotein and increasing high-density lipoprotein cholesterol) contributes to cardiovascular protection, it does not fully account for the protective effect, particularly in the application of physical therapy, including low frequency electrical stimulation. Methods: The aim of this study was to demonstrate the inhibition of stressors, such as endothelin-1 (ET-1), serotonin (5-hydroxytryptamine, 5-HT), prostaglandin $F2\alpha$ ($PGF2\alpha$), and a protein kinase C (PKC) activator 12-deoxyphorbol 13-isobutyrate (DPB), induced isometric tension by $17\beta$-estradiol in vascular smooth muscle strips, respectively. In addition, the effects of low frequency electrical stimulation at the meridian points (CV-3, -4, Ki-12, SP-6, LR-3, BL-25, -28, -32, -52) on the indirect antihypertensive effect were examined by monitoring the changes in the serum $17\beta$-estradiol concentration in healthy volunteers. Results: Isometric tension analysis showed that the responses of inhibited tension by $17\beta$-estradiol were similar to the same stressors in rat aortic smooth muscle strips. Furthermore, although the continued amplitude modulation (AM) type of electrical stimulation was not increased significantly by electrical stimulation, the current of the frequency modulation (FM) type of low frequency electrical stimulation increased the serum $17\beta$-estradiol concentration in normal volunteers. Conclusion: These results, in part, suggest that $17\beta$-estradiol has the capacity to supress stressor-induced muscle tension, and electrical stimulation, particularly current of the FM type, has a modulatory effect on the sex steroid hormones, particularly $17\beta$-estradiol, in healthy volunteers.

• 한국수정란이식학회지
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• 제24권1호
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• pp.1-14
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• 2009

Ovarian cancer is a significant cause of cancer-related death in women, but the main biological causes remain open questions. Hormonal factors have been considered to be an important determinant causing ovarian cancer. Recent studies have shown that gonadotropin-releasing hormone (GnRH)-I and its analogs have clinically therapeutic value in the treatment of ovarian cancer. In addition, numerous studies have shown that the potential of GnRH-II in normal reproductive system or reproductive disorder. GnRH-I receptors have been detected in approximately 80% of ovarian cancer biopsy specimens as well as normal ovarian epithelial cells and immortalized ovarian surface epithelium cells. GnRH-II receptors have also been found to be more widely expressed than GnRH-I receptors in mammals, suggesting that GnRH receptors may have additional functions in reproductive system including ovarian cancer. The signal transduction pathway following the binding of GnRH to GnRH receptor has been extensively studied. The activation of protein kinase A/C (PKA/PKC) pathway is involved in the GnRH-I induced anti-proliferative effect in ovarian cancer cells. In addition, GnRH-I induced mitogen-activated protein kinase (MAPK) activation plays a role in anti-proliferative effect and apoptosis in ovarian cancer cells and the activation of transcriptional factors related to cellular responses. However, the role of GnRH-I and II receptors, there are discrepancies between previous reports. In this review, the role of GnRH in ovarian cancer and the mechanisms to induce anti-proliferation were evaluated.