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      • KCI등재후보

        A Novel Simple Method to Purify Recombinant Soluble Human Complement Receptor Type 1 (sCR1) from CHO Cell Culture

        Pi-Chao Wang,Hisamune Kato,Takehiro Inoue,Noriyuki Ishii,Yoshinobu Murakami,Masatoshi Matsumura,Tsukasa Seya 한국생물공학회 2002 Biotechnology and Bioprocess Engineering Vol.7 No.2

        The human complement receptor type 1 (CR1, C3 b/C4b receptor) is a polymorphic membrane glycoprotein expressed on human erythrocytes, peripheral leukocytes, plasma and renal glomerular podocytes, which consists of transmembrane and cytoplasmic domains with 30 repeating homologous protein domains known as short consensus repeats (SCR). CR1 has been used as an inhibitor for inflammatory and immune system for the past several years. Recently, it is reported that CR1 was found to suppress the hyper-acute rejection in xeno-transplantation and can be used to cure autoimmune diseases. A soluble form of CR1, called sCR1, is a recombinant CR1 by cleaving the transmembrane domain at C-terminus and has been expressed in Chinese Hamster Ovary (CHO) cells. Several purification methods for sCR1 from CHO cells have been reported, but most of them require complicated steps at high cost. Moreover, such methods are mostly performed under the pH condition apt to denaturing sCR1 and causes sCR1 losing its activity. We here report a rapid and efficient method to purify sCR1 from CHO cell. The new method consists of a two-stage of cell culture by cultivating cells in serum medium followed by serum-free medium, and a two-stage of column purification by means of heparin and gel filtration column chromatography. By using this novel method, sCR1 can be purified in a simple and effective way with high yield and purity. Furthermore, the purified sCR1 was confirmed to retain its activity to suppress the complement activation in vivo and ex vivo.

      • KCI등재후보
      • KCI등재

        BRCA1/2 mutation status in patients with metachronous breast and ovarian malignancies: clues towards the implementation of genetic counseling

        Angel Chao,Yi-Hao Lin,Lan-Yan Yang,Ren-Chin Wu,Wei-Yang Chang,Pi-Yueh Chang,Shih-Cheng Chang,Chiao-Yun Lin,Huei-Jean Huang,Cheng-Tao Lin,Hung-Hsueh Chou,Kuan-Gen Huang,Wen-Ling Kuo,Ting-Chang Chang,Ch 대한부인종양학회 2020 Journal of Gynecologic Oncology Vol.31 No.3

        Objective: The characteristics of patients with metachronous breast and ovarian malignancies and the pathogenic role of BRCA1/2 mutations remain poorly understood. We investigated these issues through a review of hospital records and nationwide Taiwanese registry data, followed by BRCA1/2 mutation analysis in hospital-based cases. Methods: We retrospectively retrieved consecutive clinical records of Taiwanese patients who presented with these malignancies to our hospital between 2001 and 2017. We also collected information from the Data Science Center of the Taiwan Cancer Registry (TCR) between 2007 and 2015. Next-generation sequencing and multiplex ligation-dependent probe amplification were used to identify BRCA1/2 mutations and large genomic rearrangements, respectively. When BRCA1/2 mutations were identified in index cases, pedigrees were reconstructed and genetic testing was offered to family members. Results: A total of 12,769 patients with breast cancer and 1,537 with ovarian cancer were retrieved from our hospital records. Of them, 28 had metachronous breast and ovarian malignancies. We also identified 113 cases from the TCR dataset. Eighteen hospital-based cases underwent BRCA1/2 sequencing and germline pathogenic mutations were detected in 7 patients (38.9%, 5 in BRCA1 and 2 in BRCA2). All BRCA1/2 mutation carriers had ovarian high-grade serous carcinomas. Of the 12 patients who were alive at the time of analysis, 5 were BRCA1/2 mutation carriers. All of them had family members with BRCA1/2-associated malignancies. Conclusions: Our results provide pilot evidence that BRCA1/2 mutations are common in Taiwanese patients with metachronous breast and ovarian malignancies, supporting the clinical utility of genetic counseling.

      • KCI등재

        Plasma metabolites associated with physiological and biochemical indexes indicate the effect of caging stress on mallard ducks (Anas platyrhynchos)

        Zheng Chao,Wu Yan,Liang Zhen Hua,Pi Jin Song,Cheng Shi Bin,Wei Wen Zhuo,Liu Jing Bo,Lu Li Zhi,Zhang Hao 아세아·태평양축산학회 2022 Animal Bioscience Vol.35 No.2

        Objective: Cage rearing has critical implications for the laying duck industry because it is convenient for feeding and management. However, caging stress is a type of chronic stress that induces maladaptation. Environmental stress responses have been extensively studied, but no detailed information is available about the comprehensive changes in plasma metabolites at different stages of caging stress in ducks. We designed this experiment to analyze the effects of caging stress on performance parameters and oxidative stress indexes in ducks. Methods: Liquid chromatography tandem mass spectrometry (LC/MS-MS) was used to determine the changes in metabolites in duck plasma at 5 (CR5), 10 (CR10), and 15 (CR15) days after cage rearing and traditional breeding (TB). The associated pathways of differentially altered metabolites were analyzed using Kyoto encyclopedia of genes and genomes (KEGG) database. Results: The results of this study indicate that caging stress decreased performance parameters, and the plasma total superoxide dismutase levels were increased in the CR10 group compared with the other groups. In addition, 1,431 metabolites were detected. Compared with the TB group, 134, 381, and 190 differentially produced metabolites were identified in the CR5, CR10, and CR15 groups, respectively. The results of principal component analysis (PCA) show that the selected components sufficiently distinguish the TB group and CR10 group. KEGG analysis results revealed that the differentially altered metabolites in duck plasma from the CR5 and TB groups were mainly associated with ovarian steroidogenesis, biosynthesis of unsaturated fatty acids, and phenylalanine metabolism. Conclusion: In this study, the production performance, blood indexes, number of metabolites and PCA were compared to determine effect of the caging stress stage on ducks. We inferred from the experimental results that caging-stressed ducks were in the sensitive phase in the first 5 days after caging, caging for approximately 10 days was an important transition phase, and then the duck continually adapted. Objective: Cage rearing has critical implications for the laying duck industry because it is convenient for feeding and management. However, caging stress is a type of chronic stress that induces maladaptation. Environmental stress responses have been extensively studied, but no detailed information is available about the comprehensive changes in plasma metabolites at different stages of caging stress in ducks. We designed this experiment to analyze the effects of caging stress on performance parameters and oxidative stress indexes in ducks.Methods: Liquid chromatography tandem mass spectrometry (LC/MS-MS) was used to determine the changes in metabolites in duck plasma at 5 (CR5), 10 (CR10), and 15 (CR15) days after cage rearing and traditional breeding (TB). The associated pathways of differentially altered metabolites were analyzed using Kyoto encyclopedia of genes and genomes (KEGG) database.Results: The results of this study indicate that caging stress decreased performance parameters, and the plasma total superoxide dismutase levels were increased in the CR10 group compared with the other groups. In addition, 1,431 metabolites were detected. Compared with the TB group, 134, 381, and 190 differentially produced metabolites were identified in the CR5, CR10, and CR15 groups, respectively. The results of principal component analysis (PCA) show that the selected components sufficiently distinguish the TB group and CR10 group. KEGG analysis results revealed that the differentially altered metabolites in duck plasma from the CR5 and TB groups were mainly associated with ovarian steroidogenesis, biosynthesis of unsaturated fatty acids, and phenylalanine metabolism.Conclusion: In this study, the production performance, blood indexes, number of metabolites and PCA were compared to determine effect of the caging stress stage on ducks. We inferred from the experimental results that caging-stressed ducks were in the sensitive phase in the first 5 days after caging, caging for approximately 10 days was an important transition phase, and then the duck continually adapted.

      • Supplementing lactation diets with herbal extract mixture during summer improves the performance of sows and nursing piglets

        Liu, Wen-Chao,Yun, Hyeok-Min,Pi, Seung-Ho,Kim, In-Ho De Gruyter Open 2017 Annals of animal science Vol.17 No.3

        <P>A total of 45 Landrace x Yorkshire multiparous sows were used to evaluate the effect of dietary herbal extract mixture (Scutellaria baicalensis and Lonicera japonica, HEM) supplementation in lactating sows under heat stress. Sows were randomly allotted to 1 of 3 dietary treatments: 1) CON, basal diet; 2) TRT 1, basal diet with 5 g/d HEM; 3) TRT 2, basal diet with 10 g/d HEM. During lactation, dietary HEM supplementation linearly increased (P<0.05) the average daily feed intake (ADFI) and linearly decreased (P<0.05) backfat loss. The digestibility of dry matter (DM) was increased after farrowing (linear, P<0.05; quadratic, P<0.05) and weaning (linear, P<0.05) by HEM supplementation. Furthermore, HEM treatment led to a lower (linear, P<0.01) serum cortisol level. In addition, administration of HEM improved (linear, P<0.05) the piglets weaning weight and overall average daily gain (ADG) during suckling period. Meanwhile, on day 7 and 14 after birth, the fecal score of piglets was decreased (linear, P<0.01) by HEM supplementation. Taken together, under high ambient temperatures, inclusion of HEM to lactation diets could improve the feed intake, digestibility of DM, piglets weaning weight and ADG, while decreasing backfat loss, serum cortisol level, as well as the diarrhea of piglets.</P>

      • KCI등재

        Latest research progress on anticancer effect of baicalin and its aglycone baicalein

        Lin Wang,Ting Feng,Zhilian Su,Chao Pi,Yumeng Wei,Ling Zhao 대한약학회 2022 Archives of Pharmacal Research Vol.45 No.8

        Cancer, which is a leading cause of deathsaround the world, is characterized by genetic mutationsand epigenetic changes. Baicalin and its aglycone baicalein,the major bioactive fl avones derived from the driedroot of Scutellaria baicalensis Georigi, belong to fl avonoidcompounds. Many studies demonstrated that both of themexhibited remarkable promising anticancer activities. Thisstudy summarized potential anticancer mechanisms of baicalinand baicalein including induction of apoptosis, initiationof cell cycle arrest, suppression of metastasis, induction ofautophagy, and regulation of immunity. Combination strategiesinvolving baicalin or baicalein as chemotherapeuticadjuvants, clinical trial and safety were also discussed. In addition, we compared the diff erence in their anticancereff ects. Interestingly, baicalein showed quicker and strongerinhibitory eff ects on multiple cancers than those of baicalin,probably due to its smaller size and high lipophilicity whichcontribute to fast absorption and improve ability to penetratecells. Taken together, both baicalin and baicalein are eff ectivein treating cancer with good tolerance. However, deglycosylationof baicalin to baicalein was found to have strongeranticancer potential.

      • SCIESCOPUSKCI등재

        Construction of Glomerular Epithelial Cells Expressing Both Immune Tolerance and GFP Genes and Application to Cell Therapy by Cell Transplantation

        Ohga, Masahiro,Ogura, Mariko,Matsumura, Mastoshi,Wang, Pi-Chao The Korean Society for Biotechnology and Bioengine 2002 Biotechnology and Bioprocess Engineering Vol.7 No.5

        Cell therapy applied to wound healing or tissue regeneration presents a revolutionary realm to which principles of gene engineering and delivery may be applied. One promising application is the transplantation of cells into the wounded tissue to help the tissue repair. However, when cells are transplanted from in vitro to in vivo, immune rejection occurs due to the immune response triggered by the activation of T-cell, and the transplanted cells are destroyed by the attack of activated T-cell and lose their function. Immune suppressant such as FK506 is commonly used to suppress immune rejection during transplantation. However, such kind of immune suppressants not only suppresses immune rejection in the periphery of transplanted cells but also suppresses whole immune response system against pathogenic infection. In order to solve this problem, we developed a method to protect the desired cells from immune rejection without impairing whole immune system during cell transplantation. Previously, we reported the success of constructing glomerular epithelial cells for removal of immune complex, in which complement receptor of type 1 (CR1) was over-expressed on the membrane of renal glomerular epithelial cells and could bind immune complex of DNA/anti-DNA-antibody to remove immune complex through phagocy-tosis [1]. Attempting to apply the CR1-expressing cells to cell therapy and evade immune rejection during cell transplantation, we constructed three plasmids containing genes encoding a soluble fusion protein of cytolytic T lymphocyte associated antigen-4 (CTLA4Ig) and an enhanced green fluorescent protein (EGFP). The plasmids were transfected to the above-mentioned glomerular epithelial cells to express both genes simultaneously. Using the clone cells for cell transplantation showed that mice with autoimmune disease prolonged their life significantly as compared with the control mice, and two injections of the cells at the beginning of two weeks resulted in remarkable survivability, whereas it requires half a year and 50 administrations of proteins purified from the same amount of cells to achieve the same effect.

      • KCI등재

        Initial Factors Influencing Duration of Hospital Stay in Adult Patients With Peritonsillar Abscess

        Yu-Hsi Liu,Hsing-Hao Su,Yi-Wen Tsai,Yu-Yi Hou,Kuo-Ping Chang,Chao-Chuan Chi,Ming-Yee Lin,Pi-Hsiung Wu 대한이비인후과학회 2017 Clinical and Experimental Otorhinolaryngology Vol.10 No.1

        Objectives. To review cases of peritonsillar abscess and investigate the initial clinical factors that may influence the duration of hospitalization. To determine the predictive factors of prolonged hospital stay in adult patients with peritonsillar abscess. Methods. Subjects were adults hospitalized with peritonsillar abscess. We retrospectively reviewed 377 medical records from 1990 to 2013 in a tertiary medical center in southern Taiwan. The association between clinical characteristics and the length of hospital stay was analyzed with independent t-test, univariate linear regression and multiple linear regression analysis. Results. The mean duration of hospitalization was 6.2±6.0 days. With univariate linear regression, a prolonged hospital stay was associated with several variables, including female gender, older ages, nonsmoking status, diabetes mellitus, hypertension, band forms in white blood cell (WBC) counts, and lower hemoglobin levels. With multiple linear regression analysis, four independent predictors of hospital stay were noted: years of age (P<0.001), history of diabetes mellitus (P<0.001), ratio of band form WBC (P<0.001), and hemoglobin levels (P<0.001). Conclusion. In adult patients with peritonsillar abscess, older ages, history of diabetes mellitus, band forms in WBC counts and lower hemoglobin levels were independent predictors of longer hospitalization.

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