• Journal of Physiology & Pathology in Korean Medicine
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• 제23권1호
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• pp.84-92
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• 2009

This experiment was investigated the effects of fresh and ginger processed Pinelliae Rhizoma extracts on hair growth activity, and its fractions(chloroform, ethyl acetate and water fractions) obtained from fresh Pinelliae Rhizoma on hair growth activity of the normal and spontaneous alopecia areata model of C57BL/6N mice for 16 days. The results were as follows: In fresh Pinelliae Rhizoma extracts treated group, hair growth effect was observed in whole skin area(100%) all the normal mice in whose hair had been clipped on 16th days. In ginger processed Pinelliae Rhizoma extracts treated group, hair growth effect was observed in whole skin area in 25% of normal mice in whose hair had been clipped on 16th days. But in control group, hair growth effect was observed in a part of whole skin area in 25% of normal mice. In fresh Pinelliae Rhizoma extracts treated group, hair follicles of middle stage of anagen phase was observed and it were grown down to subcutaneous tissue of skin in all the mice on 10th day. But in ginger processed Pinelliae Rhizoma extracts treated group and control group, Most of hair follicles of telogen phase was observed in skin. The treatment of extracts of fresh Pinelliae Rhizoma increased the expression of TGF-$\beta$(146%), IGF(107%), and prolactin(115%) in the skin of normal C57BL/6N mice compared to control group(100%). But expression of placenta lactogen(93%) was decreased in the skin of normal C57BL/6N mice compared to control group(100%). In spontaneous alopecia model, The hair growth activity of fresh Pinelliae Rhizoma extracts treated group(100%) was observed to be strong compared with the control group(20%) on 15th day. Hair growth activity on chloroform fractions of fresh Pinelliae Rhizoma extracts was observed in whole skin area in 75% of normal mice on the 9th day. In water and ethyl acetate fractions, hair growth activity was observed in a part of whole skin in 75% and 25% of normal mice, respectively. but hair growth activity of control group was not observed. After application of fractions of fresh Pinelliae Rhizoma extracts for 10 days, hair follicles of chloroform fraction treated group was observed middle stage of anagen phase and hair follicle were grown down to subcutaneous tissue of skin in all the mice. But hair follicles of initial stage of anagen phase were observed in water and ethyl acetate fractions. Most of hair follicles of telogen phase was observed in skin of control group. These experiments suggest that extracts of fresh Pinelliae Rhizoma may stimulate the topical hair growth activity and its chloroform fractions can be useful for treatment of alopecia areata.

• Horticultural Science & Technology
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• 제34권4호
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• pp.655-661
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• 2016

To develop scab-resistant pear (Pyrus spp.) varieties with fruits that are as crisp and juicy as Asian pears, a cross was made between 'Whangkeumbae' and 'Bartlett' varieties (P. pyrifolia ${\times}$ P. communis) at the Pear Research Institute of the National Institute of Horticultural & Herbal Science, Rural Development Administration, in 1994. Among the 285 seedlings, 'Greensis' was first selected in 2006 for its good eating quality and named in 2012 after regional adaptation tests in nine regions and ten experimental plots from 2007 to 2012. The tree showed a vigorous growth habit and semi-spreading characteristics, like 'Whangkeumbae'. The optimum fruit harvest date was also around Sept. 26 and fruit was round in shape and green in skin color at maturity. Average fruit weight was 470g, and the soluble solids content was $12.4^{\circ}Brix$. The flesh was very crisp and juicy, and had good eating quality. Its' leaf size was similar with 'Bartlett' and smaller than 'Whangkeumbae'. The average of full bloom date of 'Greensis' was determined as Apr. 26, which was six days later than 'Whangkeumbae' and similar with 'Bartlett'. S genotypes of 'Greensis' were identified as $S_4S_e$ by S-allele PCR product sequencing analysis. It seems that the $S_4$ allele was inherited from 'Whangkeumbae' and the Se allele from 'Bartlett'. 'Greensis' displayed strong resistance to scab disease caused by Venturia nashicola, similar to European pear cultivars like 'Beurre Hardy' and, 'Conference'. 'Greensis' was also highly resistant to black leaf spot (Alternaria kikuchiana) in the field

• Development and Reproduction
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• 제11권3호
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• pp.263-272
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• 2007

Contrast to mouse where its in vitro maturation rates are high without specific supplements or presence of the cumulus cells, there are some species, such as porcine, where its in vitro oocyte maturation rates are still very low. This comparative study was conducted to investigate the role of malate dehydrogenase(Mor2) during oocyte maturation by RNAi in the mouse and porcine. The Mor2 double-stranded RNA(dsRNA) was prepared speciesspecifically and microinjected into the cytoplasm of denuded germinal vesicle(GV) oocytes. Oocytes were cultured for 48 h(porcine) and 16 h(mouse) in M199 with 10% porcine follicular fluid, pyruvate, p-FSH, EGF, cystein, and estradiol-$17{\beta}$. We measured changes in oocyte morphology, maturation rates and mRNA levels after Mor2 RNAi. We confirmed gene sequence-specific knock down of Mor2 mRNA in both species after Mor2 RNAi. In contrast to our previous finding that mMor2 RNAi resulted in GV arrest in the mouse, we found that pMor2 RNAi resulted in MI arrest in denuded porcine oocytes(58%), but developed to MII(84.4%) in COCs. To determine whether this difference between mouse and porcine RNAi is due to differences in culture media, we cultured mouse oocytes in the M199 media for 16 h after mMor2 RNAi. Mouse oocytes were developed to MII stage(62%) and there was no statistical difference compared to that of non-injected(76.8%) and buffer-injected(73.3%) control groups. Therefore, we concluded that the mouse and porcine oocytes are having different metabolic systems in relation to malate dehydrogenase for oocyte maturation. This could be a basis for differences in maturation rates in vitro in two species. Further scrutinized studies on the metabolic pathways would led us in finding better culture system to improve oocyte maturation rates in vitro, especially in more challenging species like the porcine.

• Journal of Life Science
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• 제26권9호
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• pp.1056-1062
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• 2016

Lung cancer is currently the most common malignant disease and the leading cause of mortality in the world and non-small cell lung cancer (NSCLC) accounts for 75-80% of lung cancer cases. miR-155 gene was found to be over expressed in several solid tumors, such as thyroid carcinoma, breast cancer, colon cancer, cervical cancer, pancreatic ductal adenocarcinoma (PDAC) and lung cancer. The aims of this study were to define the expression of miR-155 in lung cancer and its associated clinic-pathologic characteristics. Total RNA was purified from formalin-fixed, paraffin-embedded NSCLC tissues and benign lung tissues. Expression of miR-155 in human lung cancer tissues were evaluated as mean fold changes of miR-155 in cancer tissues compared to benign lung tissues by quantitative real-time reverse transcriptase polymerase chain reaction (real-time qRT-PCR) and associations of miR-155 expression with clinic-pathologic findings of cancer. Compared with the benign control group, miR-155 expression was significantly overexpressed in NSCLCs (p=<0.001). miR-155 was more overexpressed in squamous cell carcinoma than in adenocarcinoma. Poorly differentiated tumors showed significantly overexpression of miR-155 than well-differentiated tumors (p=<0.001). Overexpression of miR-155 was significantly associated with lymph node metastasis (p=<0.05). In survival analysis for all NSCLC patients, high miR-155 expression was significantly correlated with worse overall survival (p=<0.05). These results suggested that miR-155 might play an important role in lung cancer progression and metastasis.

• Tuberculosis and Respiratory Diseases
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• 제58권4호
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• pp.359-366
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• 2005

Background : IGFBP-3 inhibits the mitogenic and anti-apoptotic activity of IGF by blocking the binding of IGF to its receptor. However, under certain circumstances, IGFBP-3 can enhance the activity of IGF by protecting IGF from its degradation. More than half of the interindividual variations in IGFBP-3 levels are known to be genetically determined by the polymorphism at -202 locus of IGFBP-3 gene. Method : We attempted to ascertain whether A-202C polymorphic variation of IGFBP-3 gene constitutes a risk factor for non-small cell lung cancer (NSCLC), using PCR-restriction fragment length polymorphism (RFLP). Our study included 104 NSCLC patients and 104 age-, gender-, and smoking status-matched control subjects. Result : In the 104 NSCLC subjects, the genotypic frequencies at the -202 site were as follows: AA = 67 (64.4%), AC = 35 (33.7%), and CC = 2 (1.9%). We did detect significant differences in the genotypic distribution between the NSCLC and the control subjects (p<0.05), and the NSCLC risk correlated significantly with AA genotype at the -202 locus (AA>AC>CC). Using CC genotype as a reference, the odds ratio (OR) for the subjects with AC genotype was 2.60 (95% CI: 0.89 - 8.60), and the OR associated with AA genotype was 5.89 (95% CI: 1.92 - 21.16). Conclusion : These results indicate that the dysregulation of IGF axis should now be considered as another important risk factor for NSCLC, and a potential target for novel antineoplastic therapies and/or preventative strategies in high-risk groups.

• Journal of Life Science
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• 제26권6호
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• pp.689-697
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• 2016

Kefir is an acidic-alcoholic fermented milk product originating from the Caucasian mountains. Kefir has long been known for its probiotic health benefits, including its immunomodulatory effects. The objectives of this study were to investigate the properties of a fermented whey product and to examine the effects of kefir grains on the in vitro immune-modulation of human mast cell-1 (HMC-1). The results showed that the whey fermented by kefir grains contained the maximum lactic acid bacteria and yeast for 16 hr by 1.83×10⁸ and 6.5×10⁵ CFU/ml, respectively, and lactose and whey proteins were partially hydrolyzed. The experimental whey fermented by kefir grains exhibited an in vitro anti-inflammatory effect on the HMC-1 line for 8, 16, and 24 hr, and this effect induced the expression of interleukin (IL)-4 as a pro-inflammatory cytokine, but not for 48 hr by RT-PCR in HMC-1 cells. In addition, the same phenomenon was observed for the expression of IL-8 as a pro-inflammatory cytokine by the kefir-fermented whey during the same periods of 8-48 hr under the same conditions. These cytokines resulted in the production of IL-4 at 20-25 ng in HMC-1 cells for 8, 16, and 24 hr, whereas 5 ng was produced for 48 hr by the fermented whey. In contrast, IL-8 was produced at 15-20 ng in HMC-1 cells during 4, 8, 16, and 24 hr, while 7 ng was produced at 48 hr. It was concluded that the whey fermented by kefir grains possesses a potential anti-inflammatory function, which could be used for an industrial application as an ingredient of functional foods and pharmaceutical products.

• Korean Journal of Clinical Laboratory Science
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• 제50권4호
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• pp.449-456
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• 2018

Cervical cancer is the fourth most frequently diagnosed cancer in women worldwide. In lower Human Development Index countries, it has the second highest incidence and mortality among cancer in women. Therefore, better diagnosis and treatment systems are needed. Among them, estrogen receptor alpha ($ER-{\alpha}$) mRNA expression has been analyzed with RT-qPCR since several studies reported that $ER-{\alpha}$ is necessary in the maturation of the uterus and is related to cervical cancer. In this study, $ER-{\alpha}$ quantitative analysis was performed on various lesions and normal tissue samples. Based on the receiver operating characteristic (ROC) curve, its sensitivity and specificity were 85% and 75%, respectively, showing higher or similar results to those of conventional HPV tests. In addition, its expression level was analyzed with clinical information. With regression analysis, the R square value between the $ER-{\alpha}$ mRNA expression level and menopause status was 0.5041, indicating a strong correlation. This study was performed as part of a pilot study and suggests that $ER-{\alpha}$ is related to carcinogenesis. Future studies will examine other hormones and menopausal factors with a larger sample size.

• Tuberculosis and Respiratory Diseases
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• 제41권2호
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• pp.87-96
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• 1994

Background : Since tumor necrosis factor was discovered in 1975, TNF has been well known about its cytotoxic effect on tumor cells in vivo and in vitro. According to the recent improvement of molecular biological techinques, it is possible that exogenous TNF gene is transferred to tumor cells and is expressed in theirs. By virtue of TNF gene transfer, we have expected that TNF expressed in TNF-gene-transferred tumor cells would kill tumor cells in vivo without systemic side effect. The expected mechanisms in which antitumor effects of TNF expressed in TNF-gene-transferred tumor cells are working would be as followings. In the first mechanism, TNF expressed in TNF-gene-transferred tumor cells would kill tumor cells around(like homicide). In the second mechanism, TNF expressed in TNF-gene-transferred tumor cells would kill themselves(like suicide). In the third mechanism, TNF expressed in TNF-gene-transferred tumor cells would recruit immune effector cells and kill tumor cells indirectly. In the last mechanism, TNF expressed in TNF-gene-transferred tumor cells would augment cytokine such as interferon-$\gamma$ to kill tumor cells. Among these four mechanisms of antitumor effect, only the second mechanism has not been established yet. Therefore, to elucidate the second mechanism, We performed this study. Method : We transferred TNF-$\alpha$ gene to NCI-H2058, a human mesothelioma cell line and WEHI164, a murine fibrosarcoma cell line by using retroviral vector(pLT12SNTNF). And, We determined by using MTT assay whether TNF expressed in TNF-gene-transferred tumor cell lines would kill themselves like suicide or not. Then, if TNF-gene-transferred tumor cell lines would not suicide themselves, I would know more about the TNF sensitivity of TNF-gene-transferred tumor cell lines to exogenous TNF also by MTT assay. Result : NCI-H2058 and WEHI164 which were sensitive to TNF, became far less sensitive to endogenous and exogenous TNF after being transferred TNF-$\alpha$ gene to. Conclusion : TNF-gene-transfer to NCI-H2058 and WEHI164 gave them resistance to TNF.

• Radiation Oncology Journal
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• 제19권2호
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• pp.171-180
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• 2001

Purpose : Expression of TIMP, intrinsic inhibitor of MMP, is regulated by signal transduction in response to genotoxins and is likely to be an important step in metastasis, angiogenesis and wound healing after ionizing radiation. Therefore, we studied radiation mediated TIMP expression and its mechanism in head and neck cancer cell lines. Materials and Methods : Human head and neck cancer cell lines established at Asan Medical Center were used and radiosensitivity $(D_0)$, radiation cytotoxicity and metastatic potential were measured by clonogenic assay, n assay and invasion assay, respectively. The conditioned medium was prepared at 24 hours and 48 hours after 2 Gy and 10 Gy irradiation and expression of TIMP protein was measured by Elisa assay with specific antibodies against human TIMP. hTIMP1 promoter region was cloned and TIMP1 luciferase reporter vector was constructed. The reporter vector was transfected to AMC-HN-1 and -HN-9 cells with or without expression vector Ras, then the cells were exposed to radiation or PMA, PKC activator. EMSA was peformed with oligonucleotide (-59/-53 element and SP1) of TIMP1 promoter. Results : $D_0$ of HN-1, -2, -3, -5 and -9 cell lines were 1.55 Gy, 1.8 Gy, 1.5 Gt, 1.55 Gy and 2.45 Gy respectively. n assay confirmed cell viability, over $94\%$ at 24hrs, 48hrs after 2 Gy irradiation and over 73% after 10 Gy irradiation. Elisa assay confirmed that cells secreted TIMP1, 2 proteins continuously. After 2 Gy irradiation, TIMP2 secretion was decreased at 24hrs in HN-1 and HN-9 cell lines but after 10 Gy irradiation, it was increased in all cell lines. At 48hrs after irradiation, it was increased in HN-1 but decreased in HN-9 cells. But the change in TIMP secretion by RT was mild. The transcription of TIMP1 gene in HN-1 was induced by PMA but in HN-9 cell lines, it was suppressed. Wild type Ras induced the TIMP-1 transcription by 20 fold and 4 fold in HN-1 and HN-9 respectively. The binding activity to -59/-53, AP1 motif was increased by RT, but not to SP1 motif in both cell lines. Conclusions : We observed the difference of expression and activity of TIMPs between radiosensitive and radioresistant cell line and the different signal transduction pathway between in these cell lines may contribute the different radiosensitivity. Further research to investigate the radiation response and its signal pathway of TIMPs is needed.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2015년도 춘계학술대회 및 임시총회
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• pp.53-53
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• 2015

Agrobacterium tumefaciens-mediated transformation(ATMT) of Flammulina velutipes was used to produce a diverse number of transformants to discover the functions of gene that is vital for its variation color, spore pattern and cellulolytic activity. Futhermore, the transformant pool will be used as a good genetic resource for studying gene functions. Agrobacterium-mediated transformation was conducted in order to generate intentional mutants of F. velutipes strain KACC42777. Then Agrobacterium tumefaciens AGL-1 harboring pBGgHg was transformed into F. velutipes. This method is use to determine the functional gene of F. velutipes. Inverse PCR was used to insert T-DNA into the tagged chromosomal DNA segments and conducting sequence analysis of the F. velutipes. But this experiment had trouble in diverse morphological mutants because of dikaryotic nature of mushroom. It needed to make monokaryotic fruiting varients which introduced genes of compatible mating types. In this study, next generation sequencing data was generated from 28 strains of Flammulina velutipes with different phenotypes using Illumina Hiseq platform. Filtered short reads were initially aligned to the reference genome (KACC42780) to construct a SNP matrix. And then we built a phylogenetic tree based on the validated SNPs. The inferred tree represented that white- and brown- fruitbody forming strains were generally separated although three brown strains, 4103, 4028, and 4195, were grouped with white ones. This topological relationship was consistently reappeared even when we used randomly selected SNPs. Group I containing 4062, 4148, and 4195 strains and group II containing 4188, 4190, and 4194 strains formed early-divergent lineages with robust nodal supports, suggesting that they are independent groups from the members in main clades. To elucidate the distinction between white-fruitbody forming strains isolated from Korea and Japan, phylogenetic analysis was performed using their SNP data with group I members as outgroup. However, no significant genetic variation was noticed in this study. A total of 28 strains of Flammulina velutipes were analyzed to identify the genomic regions responsible for producing white-fruiting body. NGS data was yielded by using Illumina Hiseq platform. Short reads were filtered by quality score and read length were mapped on the reference genome (KACC42780). Between the white- and brown fruitbody forming strains. There is a high possibility that SNPs can be detected among the white strains as homozygous because white phenotype is recessive in F. velutipes. Thus, we constructed SNP matrix within 8 white strains. SNPs discovered between mono3 and mono19, the parental monokaryotic strains of 4210 strain (white), were excluded from the candidate. If the genotypes of SNPs detected between white and brown strains were identical with those in mono3 and mono19 strains, they were included in candidate as a priority. As a result, if more than 5 candidates SNPs were localized in single gene, we regarded as they are possibly related to the white color. In F. velutipes genome, chr01, chr04, chr07,chr11 regions were identified to be associated with white fruitbody forming. White and Brown Fruitbody strains can be used as an identification marker for F. veluipes. We can develop some molecular markers to identify colored strains and discriminate national white varieties against Japanese ones.