• 대한한의학방제학회지
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• 제31권1호
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• pp.1-10
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• 2023

Objectives : The main aim of this study was to examine the LC-MS/MS used to identify phenolic compounds of CRE(Coptidis Rhizoma 70% EtOH Extract). Also, we investigated antioxidative activities and Anti-inflammatory activities. Methods : LC-MS/MS Analysis HPLC and LC-MS/MS were performed on a 1260 series HPLC system (Agilent Technologies, Inc., California, USA) and 3200 QTrap tandem mass system (Sciex LLC) operated in positive ion mode (spray voltage set at -4.5 kV). The solvent used was DW and Acetonitrile containing 0.1% formic acid, a gradient system was used at a flow rate of 0.5 mL/min for analysis, and a Prontosil C18 column (length, 250 mm; inner diameter, 4.6 mm; particle size, 5 ㎛; Phenomenex Co., Ltd., California, USA, Biochoff Chromatography) was used. The solvent conditions used in the mobile phases were 0-10 min at 10-15% B, 10-20 min at 20% B, 20-30 min at 25%, 30-40 min at 40%, 40-50 min at 70%, 50-60 min at 95%, and 60-70 min at 95%. The analysis was performed at a wavelength of 284 nm and a temperature of 35℃. The cell viability was measured using a 3-(4,5-dimethyethiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity. We examined the effects of CRE on the lipopolysaccharide (LPS)-induced production of nitric oxide (NO) in a RAW 264.7 cells Results : The chemical analysis CRE by Liquid chromatography-tandem mass spectrometry (LC-MS/MS) confirmed that Rosmarinic acid, Ferrulic acid, 3-O-feruloylquinic acid, and 5-O-feruloylquinic acid as phenolic components. DPPH radical scavenging activity was the inhibitory activity of CRE showed at 200 ㎍/mL a statistically significant level. MTT assay demonstrated that the CRE did not have a cytotoxic effect in RAW 264.7 and LPS-induced RAW264.7 cells. Also, CRE reduced NO production in RAW 264.7 cells stimulated with LPS. Conclusions : Based on these findings, The chemical analysis 4 major components CRE such as Rosmarinic acid, Ferrulic acid, 3-O-feruloylquinic acid, and 5-O-feruloylquinic acid. Moreover, we confirmed that CRE has effects antioxidant and anti-inflammatory. The results demonstrate that CRE can be used as an antioxidant and a powerful chemopreventive ingredient against inflammatory diseases.

• Journal of Acupuncture Research
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• 제23권6호
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• pp.239-254
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• 2006

The 3rd Japan-Korea Workshop on Acupuncture and EBM was held at Kanazawa on June $16^{th}$. From Korea team, 4 papers were presented. Dr. Hahn introduced a new approach of data analysis on series of n-of-1 trials using the Bayesian statistics. It offered important information for the future n-of-1 trials. Dr. Park clearly demonstrated the significance of various sham devices proposed and stressed the importance of research questions when we choose the control intervention in RCT. Dr. Lee reported the results of survey in Korean Medical Doctors (KMD) for their point selection and techniques to the distal and local points. Dr. Kim presented the results of face to face survey on the KMD with 28 items for acupuncture treatment on the knee OA. Finally, a draft of protocol was introduced by Dr. Kim. The title was "multi-center, a randomized, single blinded, two arms, parallel-group study to compare the effectiveness and safety of 'individualized acupuncture' and 'standardized minimal acupuncture' in Korean and Japanese patients with knee osteoarthritis (Phase IV)". From Japan team, 7 speakers presented their comments and proposals on the protocol. Dr. Takahashi introduced several issues regarding n-of-1 trials and pointed out the importance of obtaining generalizability from n-of-1 trials. Dr. Shichidou pointed the importance of research design, selection of outcome measures and reduction of biases. Dr. Itoh presented the results of point selection for the knee OA based on the literature survey. Dr. Sumiya introduced several differences between KMD and Japanese acupuncturists based on the questionnaire used in KMD survey. Dr. Furuya demonstrated a result of press tack needle and its sham device on shoulder stiffness. Dr. Yamashita introduced the results of literature survey regarding adverse events occurred by acupuncture on knee OA. Dr.Tsukayama stressed the importance of responsibility of Institutional Review Board (IRB) for the conduction of clinical trials. After several issues were discussed, the need of continued meeting for final protocol development was agreed, then the workshop was closed.

• 대한한의학원전학회지
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• 제29권4호
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• pp.37-52
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• 2016

Objectives : The paper seeks to delve into a study of smallpox, specifically as one of the most representative contagious diseases during the Chosun era and how it was treated during that time. The lack of traditional medical texts that deal with smallpox has led this paper to dive into Duchanggyeongheombang. Methods : Everything related to edition and copy, and the author was organized based on pre-existing studies. The copy that is assumed as the original copy was considered and studied. The book was compared to and reviewed with Donguibogam, and the differences between the book and Korean translation were given attention as well. Results : Park Jin-hee is assumed to have authored Duchanggyeongheombang, and it is like that was had access to texts published by the state, one of which includes Donguibogam. He utilized the periods with smallpox outbreaks as his frame of study and put together a series of treatment for different symptoms. To this, he attached various medical charts, and added Korean translation for higher usability. In his book, he placed emphasis on spleen and stomach, but when needed, he did not shy away from using cold and coolness herbs. Conclusions : Duchanggyeongheombang was influenced partially by Donguibogam, but the book was given new life through the author Park Jin-hee's edition. Medical charts were added to increase its usability, and Korean translation was provided for wider audience. Its scope of prescription is very wide, and it provided the reasonings behind its medical judgements based on actual clinical studies instead of being tied down to traditions or taboos.

• Journal of Microbiology and Biotechnology
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• 제14권4호
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• pp.789-795
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• 2004

A promoter-probe shuttle vector pSK1Cat was constructed for the isolation of transcriptional signal sequences from Corynebacterium glutamicum. Besides conferring resistance to kanamycin in Escherichia coli and C. glutamicum, the vector carried a promoterless cat gene to confer resistance to chloramphenicol upon insertion of the appropriate transcriptional signals in the multiple cloning site. By utilizing the vector, a series of transcriptionally active fragments were isolated from the genome of C. glutamicum. The clones, ranging from 200 bp to 1 kb in size, were grouped into 3 classes of strong, medium, and weak, based on the chloramphenicol acetyltransferase (CAT) activity and sensitivity to the chloramphenicol of the clone-carrying C. glutamicum cells. C. glutamicum cells carrying the $P_{19}$ clone, a representative in the strong class, were able to grow on minimal agar plates containing over $40 mg/mell$ chloramphenicol, and showed CAT activity of 10 m㏖/mgㆍmin, performing slightly better than the cells carrying $P_{tac}$ , a strong E. coli promoter. Subcloning analysis of the $P_{19}$ clone identified a 180 bp intergenic fragment ($P_{180}$), which was located upstream of a gene encoding a hypothetical membrane protein. The expression conferred by $P_{180}$ was not affected by either the kinds of carbon sources or changes in temperature. These properties make the $P_{180}$ clone useful for the deregulated expression of biosynthetic genes in C. glutamicum during amino acid fermentation.

• Korean Journal of Acupuncture
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• 제23권1호
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• pp.111-118
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• 2006

Objectives : To test the therapeutic efficacy of acupuncture applied to PC6 and ST36 for the relief of fullness of epigastrium or epigastric pain. Subjects and Methods : We enrolled fifty patients in a case series study The patients were mainly female (94 %) with an average age of 55 years $({\pm}15.0)$; the average age of the male patients was 51 years $({\pm}20.0)$. Twenty four percent of patients were in acute phase which epigastric pain had lasted below 1 month. Twenty eight percent of patients were in chronic phase which the pain had lasted over 6 month. The rest were in subacute phase. Acupuncture needles were inserted into PC6 and ST36 bilaterally. The acupuncture treatments were performed 2 times a week for 4 weeks. One treat session was lasted for 30 min. At each visit, overall symptomatic improvement, frequency and intensity of symptoms were checked on a 4 scaled questionnaire. Results : After acupuncture treatment, a progressive improvement of pain intensity was reported in 46 of the 50 patients, including 23 of excellent and 23 of good. The progress of symptom was more superior in subacute phase to acute or chronic phase. The symptom of 22 of 24 patients in subacute phase was improved. No patient experienced minor side-effects during acupuncture treatment. Conclusion : These preliminary data suggest the acupuncture treatment provides good pain relief for most patients presenting with epigastric pain. Randomized trials with appropriate control groups are needed to validate the effectiveness of this therapy in the treatment of epigastric pain.

• Archives of Pharmacal Research
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• 제23권5호
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• pp.525-530
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• 2000

mST3GaIV synthesizes ganglioside GM3, the precursor for simple and complex a- and b- series gangliosides, and the expression and regulation of mST3GaIV (CMP-NeuAc: lactosylceramide $\alpha$2,3-sialyltransferase) activity is central to the production of almost all gangliosides, a class of glycosphingolipids implicated in variety of cellular processes such as transmembrane signaling, synaptic transmission, specialized membrane domain formation and cell-cell interactions. To understand the developmental expression of mST3GaIV in mice, we investigated the spatial and temporal expression of mST3GaIV mRNA during the mouse embryogenesis [embryonic (E) days; 19, E11, E13, E15] by in situ hybridization with digoxigenin-labeled RNA probes. All tissues from 19 and E11 were positive for mST3GaIV mRNA. On E13, mST3GaIV mRNA was expressed in various neural and non-neural tissues. In contrast to these, on E15, the telencephalon and liver produced a strong expression of mST3GaIV which was a quite similar to that of E13. In this stage, mST3GaIV mRNA was also expressed in some non-neural tissues. These data indicate that mST3GaIV is differently expressed at developmental stages of embryo, and this may be importantly related with regulation of organogenesis in mice.

• 생명과학회지
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• 제19권2호
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• pp.163-168
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• 2009

천초근은 전통적으로 동양의학에서 항암제로 사용되어왔는데 인간 급성 백혈병 세포주인 Jurkat T 세포를 사용하여 천초근의 세포독성기작을 알아보았다. 천초근 뿌리(3 kg)를 메탄올로 추출, 증류한 후 물에 녹여 다시 dichloromethane으로 추출분획 하였다. 세포독성 활성을 보이는 dichloromethane 추출물을 연속적으로 HPLC를 통해 분리하였고 그 활성물질(65 mg)을 CCH1이라 명명하였다. CCH1을 0.5 ${\mu}g$/ml에서 2.0 ${\mu}g$/ml의 농도로 처리하고 세포사멸 과정을 보았다. 즉, mitochondria cytochrome c 방출, casapase-8, -9 및 caspase-3의 활성화, PARP 분해, DNA 단편화 현상들이 일어나는 것을 관찰하였다. 하지만, mitochondria cytochrome c 방출 억제자인 Bcl-xL이 과발현되는 Jurkat T 세포에서는 세포사멸현상이 일어나지 않았다. 이러한 결과는 CCH1이 mitochondria 의존적인 신호전달 과정을 통해서 세포사멸을 유도 한다고 할 수 있다. 그리고 CCH1에 의한 세포독성은 혈액에서 분리한 단핵구 세포보다 Jurkat T 세포에서 보다 강한 활성을 보였다.

• 한국동물학회지
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• 제29권3호
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• pp.201-207
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• 1986

소금쟁이 (Gerris paludum insuralis)성체의 호흡률을 $25^\\circ C, 30^\\circ C, 35^\\circ C$ 및 $40^\\circ C$에서 암, 수별로 각각 1시간동안 매 5분간격으로 산소소비율을 측정하여 다음과 같은 결론을 얻었다. (1) $30^\\circ C$에서는 시간경과에 따른 산소소비량의 변화가 다른 온도군에 비해 매우 적었으며, 첫 5분간과 마지막 5분간의 양이 거의 비슷하여 생활적온으로 판단되며, 그 외의 온도에서는 시간경과에 따라 단위시간당 산소소비량은 주기적인 감소상을 나타내었다. (2) 산소소비율은 대체로 수컷이 암컷보다 많았다. (3) 산소소비량은 $25^\\circ C, 30^\\circ C, 35^\\circ C$ 및 $40^\\circ C$에서 각각 0.42, 0.49, 0.79 및 1.11 $\\mu$l/mg/hr로서 온도가 높아짐에 따라 증가하였다. (4) $Q_10$은 수컷에서는 $25^\\circ C \\sim 35^\\circ C$의 온도간격에서 높았고, 암컷은 $30^\\circ C \\sim 40^\\circ C$의 온도간격에서 높았다. 평균 $Q_10$은 2.12였다. (5) 생체량 10 mg, 건조중량 5 mg이상에서는 체중이 높을수록 산소소비량은 감소하였다. (6) 호흡정지고온과 호흡정지저온은 각각 $48.31^\\circ C$와 $17.68^\\circ C$였다. (7) $30^\\circ C$를 제외한 세 온도군에서 호흡이 정지되는 시간은 약 110분 후로 추정되었다.

• Nutrition Research and Practice
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• 제8권5호
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• pp.509-515
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• 2014

BACKGROUND/OBJECTIVES: The root of Vitis amurensis Ruprecht, a sort of wild-growing grape, has been used in oriental medicine for treatment of skin ailments; however, its dermatological activity is not sufficiently understood. The aim of this study was to investigate tyrosinase inhibitory and anti-melanogenic activities of V. amurensis Ruprecht root methanol extract (VARM) in B16F10 mouse melanoma cells and to attempt to isolate and identify the active compound issued from VARM. MATERIALS/METHODS: Anti-melanogenic activity of VARM was analyzed in ${\alpha}$-melanocyte stimulating hormone (MSH)-stimulated B16F10 cells through evaluation of antioxidative activity as well as inhibited tyrosinase activity and melanin contents compared with those of kojic acid and arbutin. After anti-melanogenic analysis of VARM, serial fractionation, nuclear magnetic resonance (NMR), and thin layer chromatorgraphy (TLC) were applied for identification of active compounds contained in VARM. RESULTS: VARM significantly inhibited oxidative stress and tyrosinase activity and attenuated ${\alpha}$-MSH-induced melanin production in B16F10 cells. For isolation of active compounds, VARM was fractionated using a series of organic solvents, including dichloromethane ($CH_2Cl_2$), ethyl acetate (EtOAc), and n-butanol (n-BuOH). Among fractions showing anti-melanogenic activity, the CH2Cl2 fraction induced the most potent attenuation of melanogenesis without cytotoxicity and the major compound in the $CH_2Cl_2$ fraction was identified as betulinic acid. Betulinic acid isolated from the $CH_2Cl_2$ fraction of VARM significantly attenuated ${\alpha}$-MSH-induced melanogenesis in a dose dependent manner, which was stronger than that of arbutin used as a positive control. CONCLUSIONS: These results indicate that VARM inhibits oxidative stress, tyrosinase activity, and ${\alpha}$-MSH-induced melanogenesis in B16F10 cells, due primarily to the active compound, betulinic acid, in the $CH_2Cl_2$ fraction.

• The Korean Journal of Physiology and Pharmacology
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• 제11권5호
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• pp.175-182
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• 2007

Members of prostaglandin(PG) E-series elicit cellular effects mainly through adenylyl cyclase-cAMP signaling. The role of $PGE_2$-induced increase in cAMP has been shown to be compartmentalized in the cardiac myocytes: $PGE_2$-induced increase of cAMP is not involved in the control of cardiomyocytic contraction. The purpose of the present study was to define the effect of $PGE_1$ on the cGMP levels and the role of $PGE_1$ in the atrial secretory function. Experiments were performed in perfused beating rabbit atria and atrial contractile responses, cGMP and cAMP efflux, and atrial natriuretic peptide(ANP) secretion were measured. $PGE_1$ increased cGMP as well as cAMP efflux concentration in a concentration-dependent manner, however, no significant changes in atrial secretory responses were observed(with $1.0{\mu}M\;PGE_1$; for cGMP, $144.76{\pm}37.5%$, n=11 versus $-16.81{\pm}4.76%$, n=6, control, p<0.01; for cAMP, $187.60{\pm}41.52%$, n=11 versus $7.38{\pm}19.44%$, n=6, control, p<0.01). $PGE_1$ decreased atrial dynamics slightly but transiently, whereas $PGE_2$ showed similar effects but with lower potency. Isoproterenol increased atrial cAMP efflux(with 2.0 nM; $145.71{\pm}41.89$, n=5 versus $7.38{\pm}19.44%$, n=6, control, p<0.05) and mechanical dynamics and decreased ANP secretion. The $PGE_1$-induced increase in cGMP efflux showed a bell-shaped concentration-response curve. $PGE_1$-induced increase of cGMP efflux was not observed in the presence of L-NAME, an inhibitor of nitric oxide(NO) synthase, or ODQ, an inhibitor of NO-sensitive guanylyl cyclase. L-NAME and ODQ showed no significant effect on the $PGE_1$-induced transient decrease of atrial dynamics. These data indicate that $PGE_1$ increases cGMP levels via NO-soluble GC signaling in the cardiac atrium and also show that $PGE_1$-induced increases in cGMP and cAMP levels are not involved in the regulation of atrial secretory and contractile functions.