• 대한바이러스학회지
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• 제28권4호
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• pp.337-345
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• 1998

Hantaviruses are distributed in rodent population world-widely even in geographical areas where hemorrhagic fever with renal syndrome (HFRS) has not been reported. Various species of Family Muridae and Arvicolidae serve as the natural reservoirs of hantaviruses. Hantaan virus, Seoul virus, Puumala virus, Prospect HII virus, Sin Nombre virus and New York virus are members of genus Hantavirus and isolated from lungs of A. agrarius, R. norvegicus, C. glareolus, M. pennsylvanicus, P. maniculatus and P. leucopus respectively. This experiment was intended to find the distribution of hantavirus infection among wild rodents and isolate the hantavirus from lung tissue of seropositve Apodemus peninsulae, and compared the nucleotide and amino acid sequences with prototype of hantaan virus 76-118 strain. Hantaviral sequences were amplified from lung tissues of A. peninsulae by reverse-transcriptase polymerase chain reaction. Alignment and comparison of the 324 nucleotide of G2 region of M-genomic segment diverged 4.6% and 0% at the nucleotide and amino acid levels, and complete N protein-coding region of S-genomic segment diverged 3.7% and 1.4% nucleotide and amino acid levels, respectively. This is the report to spill-over on the hantaan virus from A. agrarius to A. peninsulae in Korea.

• The Plant Pathology Journal
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• 제25권4호
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• pp.369-375
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• 2009

The potential antiviral effects of the culture filtrates (CF) from Serratia marcescens strain Gsm01 against yellow strain of Cucumber mosaic virus (CMV-Y) were investigated. The culture filtrate of S. marcescens strain Gsm01 applied on Chenopodium amaranticolor showed high inhibitory activity, likewise no necrosis appeared when applied on the tobacco plants 2 days before CMV-Y inoculation. When plants were challenge inoculated with CMV-Y for eighteen days, the disease incidence in plants with culture filtrate of S. marcescens Gsm01 did not exceed 59%, whereas 100% of control plants were severely infected. The results of double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA), reverse transcriptase polymerase chain reaction (RT-PCR), dot blotting, and western blotting showed that culture filtrate treatment highly affected the accumulation of CMV-Y or its CP protein gene in the treated plant leaves. It was also observed that the culture filtrate had no RNase activity on genomic RNAs of CMV-Y, suggesting that culture filtrate may not contain ribosome inactivating proteins (RIPs) or proteins with RNase activity. These data shows that culture filtrate of S. marcescens strain Gsm01 seems to be a promising source of antiviral substance for the practical use.

• 대한한의학회지
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• 제21권4호
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• pp.16-25
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• 2000

Objective : Acupuncture and Radix Astragali aqua-acupuncture stimuli have long been used to cure human diseases. However, the exact physiological and biochemical mechanisms involved remain undiscovered. Thus, many attempts have been made to show the scientific mechanisms involved. The effects of acupuncture and Radix Astragali aqua-acupuncture, which was known to date, as follow; effective circulation of body blood system and proliferation of leucocytes. Methods : In this study, we have applied acupuncture and Radix Astragali aqua-acupuncture stimuli to mouse on Sinsuhyul, a stimulative point of oriental medicine, to see effects on the expression of cytokine $IL-1{\beta}$. Mice were treated with lipopolysaccharide(LPS) for inflammation induction and then reverse transcriptase-polymerase chain reaction (RT-PCR) using each primer set were performed to trace the amounts of mRNA. Results : 1. $IL-1{\beta}$ was not expressed in LPS-nontreated mice at 15 to 60 min after acupuncture-stimuli. However, expression occurred after 3hrs. 2. $IL-1{\beta}$ was specifically expressed in LPS-treated mice at 30 min after acupuncture-stimuli. 3. $IL-1{\beta}$ was expressed in LPS-nontreated mice at 30 min after Radix Astragali aqua-acupuncture stimuli, however, not expressed at 60, 180 min. 4. $IL-1{\beta}$ was gradually expressed in LPS-treated mice at 15 to 180 min after Radix Astragali aqua-acupuncture stimuli. Conclusions : $IL-1{\beta}$ in LPS-treated mice was more effective than that of LPS-nontreated mice. We are now in the process of elucidating the immunological action mechanism of acupuncture and Radix Astragali aqua-acupuncture stimuli. And cytokine $IL-1{\beta}$ can be used not only as a basis of the effects of acupuncture and Radix Astragali aqua-acupuncture but also as a diagnosis guide through the immunological actions of those.

• Journal of Microbiology and Biotechnology
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• 제13권1호
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• pp.77-84
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• 2003

Chitinase was purified from an antagonistic bacterium Bacillus sp. 7079 by ammonium sulfate precipitation, QAE-Sephadex anion exchange chromatography, Sephadex G-100 gel filtration, and SP-Sephadex cation exchange chromatography. The molecula. weight of purified chitinase (PC-1) was approximately 66.5 kDa on SDS-PACE. PC-1 exhibited optimum pH and temperature of pH 7.5 and $45^{\circ}C$, respectively. More than $80\%$ of PC-1 was stable at pH 5.0 to 9.0, and more than $90\%$ at $40^{\circ}C$. $Fe^2+\;and\;Ca^2+$ inhibited the chitinase activity about $20\%$, and EDTA and p-CMB by about $30\%$, whereas $Ag^+$ inhibited the activity up to $65\%$. The $K_m$ value of PC-1 was 1.215 mg/ml with colloidal chitin as a substrate. We also investigated the effect of PC-1 treated chitin (PCTC) on the pro-inflammatory cytokine gene expression in macrophage RAW 264.7 cells. The expression of IL-$1{\alpha}$ and IL-$1{\beta}$ mRNA gene was investigated using reverse transcriptase polymerase chain reaction (RT-PCR). IL-$1{\alpha}$ and IL-$1{\beta}$ mRNA were induced by the treatment of PCTC and chitin only in RAW 264.7 cells. These expressions were induced as early as 2 h and sustained up to 24 h in RAW 264.7 cells. IL-$1{\alpha}$ and IL-$1{\beta}$ mRNA were more strongly expressed by the treatment of PCTC than chitin treatment alone in RAW 264.7 cells.

• The Korean Journal of Physiology and Pharmacology
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• 제1권2호
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• pp.151-159
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• 1997

In an attempt to investigate whether hemorrhage affects the gene expression of the renin-angioteusin system (RAS) components in the brain and peripheral angiotensin-generating tissues, changes in mRNA levels of the RAS components in response to hemorrhage were measured in conscious unrestrained rats. Wistar rats were bled at a rate of 3 ml/kg/min for 5 min, and then decapitated 7 h after hemorrhage. Levels of mRNA for renin, angiotensinogen and angiotensin $II-AT_1$ receptor subtypes ($AT_{1A}$ and $AT_{1B}$) were determined with the methods of northern blot and reverse transcriptase-polymerase chain reaction (RT-PCR). Hemorrhage produced a profound hypotension with tachycardia, but blood pressure and heart rate recovered close to the basal level at 7 h. Plasma and renal renin levels were significantly increased at 7 h. Hemorrhage induced rapid upregulation of gene expression of both $AT_{1A}$ and $AT_{1B}$ receptor subtypes in the brainstem and hypothalamus, downregulation of them in the adrenal gland and liver. However, renin mRNA level increased in the brainstem, decreased in the liver, but was not changed in the hypothalamus, kidney and adrenals after hemorrhage. Angiotensinogen mRNA level was not significantly changed in any of the tissue except a slight increase in the liver. The kidney and liver did not show any significant change in gene expression of the RAS components. These results suggest that gene expression of the RAS in central and peripheral tissues are, at least in part, under independent control and the local RAS in each organ plays specific physiologic role.

• Journal of Korean Neurosurgical Society
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• 제49권1호
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• pp.13-19
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• 2011

Objective: The purpose of this study is to investigate the combined effects of ginkgo biloba extract, ginkgolide A and B and aspirin on SK-N-MC, human neuroblastoma cell viability and mRNA expression of growth associated protein43 (GAP43), Microtubule-associated protein 2 (MAP2), B-cell lymphoma2 (Bcl2) and protein53 (p53) gene in hypoxia and reperfusion condition. Methods: SK-N-MC cells were cultured with Dulbecco's Modified Eagle's Medium (DMEM) media in $37^{\circ}C$, 5% $CO_2$ incubator. The cells were cultured for 8 hours in non-glucose media and hypoxic condition and for 12 hours in normal media and $O_2$ concentration. Cell survival rate was measured with Cell Counting Kit-8 (CCK-8) reagent assay. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to estimate mRNA levels of GAP43, MAP2, Bcl2, and p53 genes. Results: The ginkgolide A and B increased viable cell number decreased in hypoxic and reperfused condition. The co-treatment of ginkgolide B with aspirin also increased the number of viable cells, however, there was no additive effect. Although there was no increase of mRNA expression of GAP43, MAP2, and Bcl2 in SK-N-MC cells with individual treatment of ginkgolide A, B or aspirin in hypoxic and reperfused condition, the co-treatment of ginkgolide A or B with aspirin significantly increased GAP43 and Bcl2 mRNA levels. In MAP2, only the co-treatment of ginkgolide A and aspirin showed increasing effect. The mRNA expression of p53 had no change in all treating conditions. Conclusion: This study suggests that the combined treatments of Ginkgo biloba extracts and aspirin increase the regeneration of neuroblastoma cells injured by hypoxia and reperfusion.

• Journal of Korean Neurosurgical Society
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• 제55권5호
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• pp.237-243
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• 2014

Objective : Symptomatic disc degeneration develops from inflammatory reactions in the annulus fibrosus (AF). Although inflammatory mediators during annular inflammation have been studied, the roles of matrix metalloproteinases (MMPs) and their inhibitors have not been fully elucidated. In this study, we evaluated the production of MMPs and tissue inhibitors of metalloproteinase (TIMPs) during annular inflammation using an in vitro co-culture system. We also examined the effect of notochordal cells on annular inflammation. Methods : Human AF (hAF) pellet was co-cultured for 48 hours with phorbol myristate acetate-stimulated macrophage-like THP-1 cells. hAF pellet and conditioned media (CM) from co-cultured cells were assayed for MMPs, TIMPs, and insulin-like growth factor (IGF)-1 levels using real-time reverse-transcriptase polymerase chain reaction and enzyem-linked immunosorbent assay. To evaluate whether notochordal cells affected MMPs or TIMPs production on annular inflammation, hAF co-cultured with notochordal cells from adult New Zealand White rabbits, were assayed. Results : MMP-1, -3, -9; and TIMP-1 levels were significantly increased in CM of hAF co-cultured with macrophage-like cells compared with hAF alone, whereas TIMP-2 and IGF-1 levels were significantly decreased (p<0.05). After macrophage exposure, hAF produced significantly more MMP-1 and -3 and less TIMP-1 and -2. Interleukin-$1{\beta}$ stimulation enhanced MMP-1 and -3 levels, and significantly diminished TIMP-2 levels. Co-culturing with rabbit notochordal cells did not significantly influence MMPs and TIMPs production or COL1A2 gene expression. Conclusion : Our results indicate that macrophage-like cells evoke annular degeneration through the regulation of major degradative enzymes and their inhibitors, produced by hAF, suggesting that the selective regulation of these enzymes provides future targets for symptomatic disc degeneration therapy.

• Asian Pacific Journal of Cancer Prevention
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• 제15권16호
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• pp.6685-6689
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• 2014

Background: Alterations in gene expression levels or mutations of tyrosine kinases are detected in some human cancers. In this study, we examined whether serine threonine tyrosine kinase 1 (STYK1)/novel oncogene with kinase domain (NOK) is overexpressed in patients with colorectal cancer. We also examined the clinical relevance of STYK1/NOK expression in cancer tissues. Materials and Methods: In tumor samples of patients with colorectal cancer and their matched non-cancerous samples, STYK1/NOK messenger RNA (mRNA) expression was analyzed by quantitative reverse transcriptase polymerase chain reaction. Associations between the expression levels of STYK1/NOK and clinicopathological characteristics of colorectal cancer were also assessed using Mann-Whitney U and Kruskal-Wallis tests. Results: Upregulation of STYK1/NOK was found in cancer tissues even at early stage of colorectal cancer compared to normal adjacent tissues. The optimal cutoff point of 0.198 the STYK1/NOK expression showed 0.78 sensitivity and 0.75 specificity for diagnosis. Overexpressed STYK1/NOK was correlated with tumor size but had no association with other clinicopathological characteristics of colorectal cancer. Conclusions: These results indicate that STYK1/NOK mRNA is widely expressed in the patients with colorectal cancer and suggest that inhibition of this molecule could potentially serve as a novel therapeutic target.

• Asian Pacific Journal of Cancer Prevention
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• 제15권6호
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• pp.2559-2563
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• 2014

Purpose: We investigated the expression of epithelial $Ca^{2+}$ channel transient receptor potential vanilloid (TRPV) 5 and 6 in non-small-cell lung cancer (NSCLC) and assessed their prognostic role in patients after surgical resection. Materials and Methods: From January 2008 to January 2009, 145 patients who had undergone surgical resection of NSCLCs were enrolled in the study. Patient clinical characteristics were retrospectively reviewed. Fresh tumor samples as well as peritumor tissues were analyzed for TRPV5/6 expression using immune-histochemistry (IHC) and quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). Patients were grouped based on their TRPV5 and TRPV6 levels in the tumor tissues, followed up after surgery, and statistically analyzed to examine the prognostic roles of TRPV5 and TRPV6 on patients' survival after surgical resection of NSCLCs. Results: Using IHC, among the 145 patients who had undergone surgical resection of NSCLCs, strong protein expression (grade${\geq}2$) of TRPV5 and TRPV6 was observed in a lower percentage of primary tumor tissues than in non-tumor tissues of same patients. Similar findigns were obtained with the RT-PCR test for mRNA levels. Decreased overall mRNA levels of TRPV5 and TRPV6 were associated with a worse overall survival rate (p=0.004 and p=0.003 respectively) and shorter recurrence-free survival (p<0.001 and p<0.001 respectively). The combining effect of TRPV5 and TRPV6 on survival was further investigated using multivariate analysis. The results showed that a combination of low expression of TRPV5 and TRPV6 could be an independent predictor of poor recurrence-free survival (p=0.002). Conclusions: Decreased expression of TRPV5/6 in tumor tissues was observed in NSCLC patients and was associated with shorter median survival time after surgical resection. Combined expression of TRPV5 and TRPV6 in tumor tissues demonstrated promising prognostic value in NSCLC patients.

• 한국산학기술학회논문지
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• 제14권8호
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• pp.3843-3849
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• 2013

이배체 단위발생 돼지 난자를 체외 배양시 배반포 형성 단계에서 FBS (우태아 혈청), BSA (우혈청 알부민), EGF(상피세포 성장인자)를 배양액에 첨가하였을 때 이배체 단위발생 에서 총세포수, 세포사멸 및 세포사멸에 관여하는 유전자의 발현 효과를 조사하고자 본 연구를 수행하였다. 0.4% BSA를 배양액에 첨가 하였을 때 2 세포기 단계 단위발생의 발달은 배반포 까지는 강화 되었다 (p<0.01). FBS 처리 시는 배반포의 세포 수는 감소시켰으나 세포 사멸률은 증가하였다(p<0.01). 하지만 EGF가 존재할 때 BSA 처리는 총 세포수를 증가 시켰다. RT-PCR의 결과에 의하면 EGF는 0.4% BSA가 존재하는 배양액에서는 Bcl-xL mRNA 발현을 증가시키고 BSA와 EGF 가 단독으로 존재 할 때는 효과가 없었다. 하지만 FBS 처리시 Bcl-xL 유전자 발현은 감소하고 Bak 유전자의 발현은 증가시킨다. 이러한 결과 세포사멸에 관여하는 유전자의 발현은 배양액의 첨가물에 따라 유의적으로 영향을 받으며, 돼지 배아의 체외 배양시 세포사멸과 초기발달에 관여함을 시사한다.