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Conditioned Media of ASC-17D Sertoli Cells Induce G1-Growth Arrest of DU145 Human Prostate Cancer Cells

  • Kang, Hyeog;Kang, Sang-Wook;Choi, Sang-Hyun;Lee, Kwang-Ho;Chun, Boe-Gwun;Min, Bon-Hong
    • BMB Reports
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    • v.31 no.5
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    • pp.508-514
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    • 1998
  • We studied the effects of ASC-17D rat Sertoli cell-conditioned media (rSCCM) on the proliferation of the DU145 prostate cancer cells. rSCCM was prepared from ASC-17D cells cultured in DMEM/F-12 serum-free media at a nonpermissive temperature of $40^{\circ}C$, which is the condition for the high expression of c1usterin. We found that rSCCM could inhibit the proliferation of DU145 cells by arresting the cell cycle in the G1 phase in a dose-dependent manner. This growth arresting activity was abolished by boiling rSCCM for 5 min. The G1 growth-inhibiting activity of rSCCM was also detected in other prostate-originated cancer cells examined (i.e., LNCaP and PC-3) but not in other cells (ASC-17D, HepG2, SK-N-SH, and NIH3T3). Western blot analysis of partially purified growth inhibiting fractions with the clusterin antibody showed that the cytostatic factor in rSCCM was not c1usterin. This cytostatic factor was semi purified by DEAE-Sepharose, ammonium sulfate precipitation, and Phenyl-Sepharose column chromatography, and was estimated to have a molecular weight of 88 kDa by Sephacryl S-300 gel filtration.

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Quantitative and Rapid Analysis of Transglutaminase Activity Using Protein Arrays in Mammalian Cells

  • Kwon, Mi-Hye;Jung, Jae-Wan;Jung, Se-Hui;Park, Jin-Young;Kim, Young-Myeong;Ha, Kwon-Soo
    • Molecules and Cells
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    • v.27 no.3
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    • pp.337-343
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    • 2009
  • We developed a novel on-chip activity assay using protein arrays for quantitative and rapid analysis of transglutaminase activity in mammalian cells. Transglutaminases are a family of $Ca^{2+}$-dependent enzymes involved in cell regulation as well as human diseases such as neurodegenerative disorders, inflammatory diseases and tumor progression. We fabricated the protein arrays by immobilizing N,N'-dimethylcasein (a substrate) on the amine surface of the arrays. We initiated transamidating reaction on the protein arrays and determined the transglutaminase activity by analyzing the fluorescence intensity of biotinylated casein. The on-chip transglutaminase activity assay was proved to be much more sensitive than the $[^3H]putrescine$-incorporation assay. We successfully applied the on-chip assay to a rapid and quantitative analysis of the transglutaminase activity in all-trans retinoic acid-treated NIH 3T3 and SH-SY5Y cells. In addition, the on-chip transglutaminase activity assay was sufficiently sensitive to determine the transglutaminase activity in eleven mammalian cell lines. Thus, this novel on-chip transglutaminase activity assay was confirmed to be a sensitive and high-throughput approach to investigating the roles of transglutaminase in cellular signaling, and, moreover, it is likely to have a strong potential for monitoring human diseases.

A Novel Mutant of Human Papillomavirus Type 18 E6E7 Fusion Gene and its Transforming Activity

  • Zhou, Zhi-Xiang;Zhao, Chen;Li, Qian-Qian;Zeng, Yi
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.17
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    • pp.7395-7399
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    • 2014
  • Background: Persistent human papillomavirus (HPV) infection, especially with high-risk types such as HPV16 and HPV18, has been identified as the primary cause of cervical cancer. E6 and E7 are the major onco-proteins of high-risk HPVs, which are consistently expressed in HPV infected tissues but absent in normal tissues and represent ideal therapeutic targets for immunotherapy of cervical cancer. Materials and Methods: In this study, the optimized fusion gene HPV18 E6E7 (HPV18 ofE6E7) was constructed according to genetic codon usage for human genes. At the same time, for safety future clinical application, a mutant of HPV18 ofE6E7 fusion gene was generated by site-directed mutagenesis at L52G for the E6 protein and C98G for the E7 protein. Results: HPV18-E6E7 mutant (HPV18 ofmE6E7) constructed in this work not only lost the transformation capability for NIH 3T3 cells and tumorigenicity in BALB/c nude mice, but also maintained very good stability and antigenicity. Conclusion: These results suggest that the mutant should undergo further study for application as a safe antigenspecific therapeutic vaccine for HPV18-associated tumors.

Effect of The activating blood flow and removing blood stasis Method to Prostatic disease (전립선(前立腺) 질환(疾患)에 대한 활혈거어법(活血祛瘀法)의 효과(效果))

  • Song, Bong-Keun;Kim, Joong-Kil;Park, Seung-Won;Song, Un-Yong;Jeong, In-Seok
    • The Journal of Internal Korean Medicine
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    • v.21 no.4
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    • pp.615-619
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    • 2000
  • Objective : The prostatic disease is characterized as relatively incurable, chronic and recurrent illness. Because of this property, we have the difficulty in treating this illness. Besides, the drug selectivity is restricted to the specific prostatic membrane, the barrier of prostate. This study was to evaluate the efficency of the activating blood flow and removing blood stasis Method to Prostatic disease. Methods : We investigated 51 prostatic patiens with NIH chronic prostatitis symptom index, digital rectal examination(DRE), expressed prostatic secretion(EPS). After investigation we treated the patients with herb which was aimed to activate the blood flow and remove the blood stasis method to prostatic disease. Results : After the treatment, symptom index score of the prostatic patients was reduced from 11.7 to 7.4. The cure rate of 51 prostatic patients was 76.5%. Also the cure rate of the patients who was treated over 4 weeks was higher than the patients treated below 4 weeks, 87.5% to 66.7% respectively. conclusions : These results indicate that oriental medical theraphy is useful enough to treat the prostatic patients. Therefore, the more research about the herb which has activating blood flow and removing blood stasis effect is necessary.

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Meta-analysis of Six Randomized Control Trials of Chemotherapy Plus Anti-HER Monoclonal Antibody for Advanced Gastric and Gastroesophageal Cancer

  • Luo, Huai-Qing;Han, Li;Jiang, Yan
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.13
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    • pp.5343-5348
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    • 2014
  • Background: A meta-analysis was performed to examine the benefit/risk ratio for the addition of anti- HER MoAbs to chemotherapy in patients with advanced gastric and gastroesophageal cancer from six randomized phase II/III trials. Materials and Methods: We searched relative trials from Pubmed, EMBASE, Cochrane library databases, China National Knowledge Infrastructure databases, Google Scholar and the NIH ClinicalTrials. Primary outcomes were overall response rate (ORR), progression-free survival (PFS), overall survival (OS). Secondary outcomes were toxicities. All analyses were performed using STATA 12.0. Results: This meta-analysis included six randomized controlled trials (RCTs) with 2, 297 patients and we demonstrated that the anti-HER MoAbs arm did have a positive effect on ORR in the anti-HER MoAbs arm (OR 1.28, 95% CI 1.00-1.64, p=0.01). There was an increasing benefit regarding OS (HR 0.74, 95% CI 0.60-0.88, p<0.05) and PFS (HR 0.72, 95% CI 0.60-0.84, p<0.05) in the anti-HER2 subgroup, but a reduction of OS (HR 1.11, 95% CI 0.87-1.36, p<0.05) and PFS (HR 1.13, 95% CI 0.98 -1.28, P<0.05) in anti-EGFR subgroup. Some grade 3-4 toxicity had a significantly higher incidence in the anti-HER MoAbs arm. There was no significant publication bias for all endpoints. Conclusions: The addition of trstuzumab MoAb to chemotherapy for gastric and gastroesophageal cancer significantly improved outcome of OS and PFS endpoints, while other MoAbs led to no improvement in results. Some adverse events were increased in anti-HER MoAbs arm compared with the control.

A Homeotic Gene, Hoxc8, Regulates the Expression of Proliferating Cell Nuclear Antigen in NIH3T3 Cell

  • Min, Hye-Hyun;Kang, Myeng-Mo;Kim, Myoung-Hee
    • Biomedical Science Letters
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    • v.13 no.3
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    • pp.239-244
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    • 2007
  • Hoxc8 is one of the homeotic developmental control genes regulating the expression of many downstream target genes, through which animal body pattern is established during embryonic development. In previous proteomics analysis, proliferating cell nuclear antigen (PCNA) which is also known as cyclin, has been implied to be regulated by Hoxc8 in F9 murine embryonic teratocarcinoma cell. When the 5' upstream region of PCNA was analyzed, it turned out to contain 20 Hox core binding sites (ATTA) in about 1.17 kbp (kilo base pairs) region ($-520{\sim}-1690$). In order to test whether this region is responsible for Hoxc8 regulation, the upstream 2.3 kbp fragment of PCNA was amplified through PCR and then cloned into the pGL3 basic vector containing a luciferase gene as a reporter. When the luciferase activity was measured in the presence of effector plasmid (pcDNA : c8) expressing murine Hoxc8, the PCNA promoter driven reporter activity was reduced. To confirm whether this reduction is due to the Hoxc8 protein, the siRNA against Hoxc8 (5'-GUA UCA GAC CUU GGA ACU A-3' and 5'-UAG UUC CAA GGU CUG AUA C-3') was prepared. Interestingly enough, siRNA treatment up regulated the luciferase activity which was down regulated by Hoxc8, indicating that Hoxc8 indeed regulates the expression of PCNA, in particular, down regulation in NIN3T3 cells. These results altogether indicate that Hoxc8 might orchestrate the pattern formation by regulating PCNA which is one of the important proteins involved in several processes such as DNA replication and methylation, chromatin remodeling, cell cycle regulation, differentiation, as well as programmed cell death.

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Blockage of the Immune Complex-triggered Transmembrane Proximity Between Complement Receptor Type 3 and Microfilaments by Staurosporine and Methyl-2,5-dihydroxycinnamate

  • Poo, Ha-Ryoung;Lee, Young-Ik;Todd, Robert F. III;Petty, Howard R.
    • BMB Reports
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    • v.31 no.1
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    • pp.64-69
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    • 1998
  • Recent studies have suggested that integrin (CR3) participates in the signal transduction pathways of certain GPI-anchored phagocytic receptors including $Fc{\gamma}RIIIB$. One consequence of this functional linkage is an inducible association between CR3 and cortical microfilaments that is triggered by $Fc{\gamma}RIIIB$ binding to immobilized immune complexes (IC). That this signaling event requires the co-expression of $Fc{\gamma}RIIIB$ with CR3 was documented by the use of NIH 3T3 transfectants expressing both CR3 and $Fc{\gamma}RIIIB$ (clone 3-23), CR3 alone (clone 3-19), and $Fc{\gamma}RIIIB$ alone (clone 3-15). Pretreatment of 3-23 cells with protein kinase inhibitors such as staurosporine and methyl 2,5-dihydroxycinnamate (MDHC) blocked IC-stimulated CR3 microfilament proximity without affecting the extent to which $Fc{\gamma}RIIIB$ constrains the lateral membrane mobility of a subset of CR3 on the cell surface (as measured in fluorescence recovery after photobleaching experiments). These data support that CR3 and $Fc{\gamma}RIIIB$ molecules are physically and functionally associated and that ligation of FcgRIIIB triggers CR3-dependent signal transduction.

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Clinical Review about Dysphagia associated with Acute Ischemic Stroke (급성 허혈성 뇌졸중에 수반된 연하장애에 관한 임상적 고찰)

  • 한명아;김동웅
    • The Journal of Korean Medicine
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    • v.22 no.3
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    • pp.42-50
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    • 2001
  • Objectives : Dysphagia is common and severe problems of acute stroke determining the prognosis of stroke only second to mental change, and results in secondary fatal complications such as aspiration pneumonia, malnutrition, dehydration, etc. Therefore, we were to investigate the clinical characteristics of dysphagia accompanied by acute ischemic stroke. Methods : We selected subjects through clinical notes retrospectively, whose main problems included dysphagia resulted from acute stroke within 72 hours from onset who were admitted to the Internal Medicine Department of Wonkwang Oriental Medicine Hospital from Jan. 2000 to Apr. 2001. We assessed the severity of dysphagia from admission to discharge using a staging method : stage 0 is normal without dysphagia, stage 1 is nearly normal except for intermittent dysphagia, stage 2 is compensated abnormal swallowing requiring adjusted diets or delayed feeding time, stage 3 is uncompensated abnormal swallowing resulted in weight loss down to 10% of initial and daily aspiration, coughing, and vomiting, stage 4 is uncompensated abnormal swallowing resulting in weight loss beyond 10% and recommended for non-oral feeding, and stage 5 is 100% non-oral feeding by L-tube, or gastrostomy or NPO state. Results : Dysphagia was improved statistically significantly from the mean stage of $3.6{\pm}0.29$ on admission to $1.88{\pm}0.32$ on discharge (P<0.05). On average $7.1{\pm}1.48$ days were required for improving more than one stage level. As patients were older and the stage of dysphagia was worse on admission, severity of dysphagia was more difficult to improve (correlation coefficiency was 0.55 and 0.77 respectively, P<0.05). Aspiration pneumonia was complicated in 13 patients of the total 25 at mean dysphagia stage of $3.36{\pm}0.37$. However, any specific values such as lesion size, lesion site, sex, age, past history and NIH Stroke Scale on admission did not affect it (P>0.05). Conclusion : Clinical course of dysphagia was determined about I week from the onset. Aspiration pneumonia was mainly complicated during oral feeding periods. If there were no improvement of dysphagia over 2-3 weeks, then non-oral feeding such as Levin tube or gastrostomy must be considered.

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Antioxidant value and Antiproliferative Efficacy of Mitragynine and a Silane Reduced Analogue

  • Goh, Teik Beng;Yian, Koh Rhun;Mordi, Mohd Nizam;Mansor, Sharif Mahsufi
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.14
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    • pp.5659-5665
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    • 2014
  • Background: To investigate the antioxidant value and anticancer functions of mitragynine (MTG) and its silane-reduced analogues (SRM) in vitro. Materials and Methods: MTG and SRM was analyzed for their reducing power ability, ABTS radical inhibition and 1,1-diphenyl-2-picryl hydrazylfree radicals scavenging activities. Furthermore, the antiproliferation efficacy was evaluated using MTT assay on K 562 and HCT116 cancer cell lines versus NIH/3T3 and CCD18-Co normal cell lines respectively. Results: SRM and MTG demonstrate moderate antioxidant value with ABTS assay (Trolox equivalent antioxidant capacity (TEAC): $2.25{\pm}0.02$ mmol trolox / mmol and $1.96{\pm}0.04$ mmol trolox / mmol respectively) and DPPH ($IC_{50}=3.75{\pm}0.04mg/mL$ and $IC_{50}=2.28{\pm}0.02mg/mL$ respectively). Both MTG and SRM demonstrate equal potency ($IC_{50}=25.20{\pm}1.53$ and $IC_{50}=22.19{\pm}1.06$ respectively) towards K 562 cell lines, comparable to control, betulinic acid (BA) ($IC_{50}24.40{\pm}1.26$). Both compounds showed concentration-dependent cytototoxicity effects and exert profound antiproliferative efficacy at concentration > $100{\mu}M$ towards HCT 116 and K 562 cancer cell lines, comparable to those of BA and 5-FU (5-Fluorouracil). Furthermore, both MTG and SRM exhibit high selectivity towards HCT 116 cell lines with selective indexes of 3.14 and 2.93 respectively compared to 5-FU (SI=0.60). Conclusions: These findings revealed that the medicinal and nutitional values of mitragynine obtained from ketum leaves that growth in tropical forest of Southeast Asia and its analogues does not limited to analgesic properties but could be promising antioxidant and anticancer or chemopreventive compounds.

COMPARISON OF IMAGE REFORMATION USING PERSONAL COMPUTER WITH CT SCAN RECONSTRUCTION (CT 스캔 영상재구성과 개인용 컴퓨터를 이용한 영상 재형성과의 비교에 관한 연구)

  • Jung Gi-Hun;Kim Eun-Kyung;Kim Sang-Joon
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.24 no.2
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    • pp.361-368
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    • 1994
  • Radiographic planning is needed for implant placement in order to determine implant length, jaw bone volume, anatomical stucture and so on. Radiographic examination includes conventional radiography, conventional tomography and CT scan. The most accurate mesurement can be obtained from CT scan. For the cross-sectional view of mandible, CT scan reconstruction is generally needed. But the cross-sectional view of mandible can be reformed by personal computer. This study was performed to examine the clinical usefulness of reformed image using personal computer in comparison with CT scan reconstructed image. CT axial slices of 4 mandibles of 4 volunteers were used. Digital imaging system was composed of Macintosh Ⅱ ci computer, high resolution Sony XC-77 CCD camera, Quick Capture frame grabber board and 'NIH Image' program. Seven reconstructed cross-sectional images within CT machine(CT group) were obtained. And seven reformed cross-sectional images(PC group) after digitization of CT axial slices into the personal computer were obtained. PC group was compared with CT group in the objective and subjective aspects. The results were as follow: 1. Measurement of mandibular height & width in both group showed insignificant difference(P>0.05). 2. Subjective assessment of the mandibular canal in both group showed insignificant difference(P>0.05). 3. Image reformation using personal computer could provide panoramic view, which could not be obtained in CT scan reconstruction.

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