• Journal of Microbiology and Biotechnology
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• 제19권12호
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• pp.1557-1564
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• 2009

Human CDX2 is known as a caudal-related homeodomain transcription factor that is expressed in the intestinal epithelium and is important in differentiation and maintenance of the intestinal epithelial cells. The caudal-related homeobox proteins bind DNA according to a helix-turn-helix structure, thereby increasing the structural stability of DNA. A cancer-tumor suppressor role for Cdx2 has been shown by a decrease in the level of the expression of Cdx2 in colorectal cancer, but the mechanism of transcriptional regulation has not been examined at the molecular level. We developed a large-scale system for expression of the recombinant, novel CDX2, in Escherichia coli. A highly purified and soluble CDX2 protein was obtained in E. coli strain BL21(DE3)RIL and a hexahistidine fusion system using Ni-NTA affinity column, anion exchange, and gel filtration chromatographies. The identity and secondary structure of the purified CDX2 protein were confirmed by MALDI-TOF MS, Western blot, and a circular dichroism analyses. In addition, we studied the DNA-binding activity of recombinant CDX2 by ELISA experiment and isolated human CDX2-binding proteins derived from rat cells by an immobilized GST-fusion method. Three CDX2-binding proteins were found in the gastric tissue, and those proteins were identified to the homeobox protein Hox-D8, LIM homeobox protein 6, and SMC1L1 protein.

• Asian Pacific Journal of Cancer Prevention
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• 제15권16호
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• pp.6673-6678
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• 2014

Background: To investigate the prognostic value of serum PIVKA-II (prothrombin induced by the absence of vitamin K or antagonist-II) in BCLC (Barcelona Clinic Liver Cancer) 0-A hepatocellular carcinoma (HCC) patients after curative resection. Materials and Methods: Preoperative sera were collected from 140 patients with BCLC 0-A HCCs undergoing curative resection during 2011-2012 in Zhongshan Hospital. Follow-up ended on November 2013. ELISA was used to detect the serum concentrations of preoperative PIVKA-II. The prognostic value of PIVKA-II and other clinicopathological factors was analyzed by the Kaplan-Meier method and the multivariate Cox proportional hazards model. Results: During follow-up, 39 of 140 patients suffered recurrence and the 1-year recurrence rate was 27.9%. The high-PIVKA-II expression group had lower 1-year time to progression (TTP) compared with the low-expression group (54.8% vs 20.2%, p<0.001). Patients with high preoperative PIVKA-II expression showed a relatively higher risk of developing postoperative recurrence than those with low expression in the low-recurrence-risk subgroups, including ${\alpha}$-fetoprotein ${\leq}400ng/mL$ (45.4% vs 16.7%; p=0.006), tumor size ${\leq}5cm$ (54.2% vs 18.1%; p<0.001), single tumor (56.0% vs 19.1%; p<0.001), absence of satellite lesions (53.3% vs 19.8%; p=0.001), absence of vascular invasion (52.6% vs 14.9%; p=0.002), and Edmondson stage I/II (60.9% vs 20.3%; p<0.001). PIVKA-II was the strongest independent prognostic factor for TTP (hazard ratio, 2.877; 95% CI 1.524-5.429; p=0.001). Conclusions: Elevated PIVKA-II is associated with early recurrence of BCLC 0-A HCC after curative resection and can be considered a novel prognostic predictor.

• Asian Pacific Journal of Cancer Prevention
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• 제15권12호
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• pp.4919-4923
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• 2014

Background: To investigate the effect of celecoxib on telomerase activity and apoptosis in a human laryngeal squamous carcinoma cell line (Hep-2 cells). Materials and Methods: The growth inhibition rate of Hep-2 cells in vitro was measured by MTT assay, and apoptosis by TUNEL assay and flow cytometry (FCM). The TRAP-ELISA method was used to determine telomerase activity in Hep-2 cells. The mRNA expression of human telomerase RNA component(hTR), human telomerase reverse transcriptase (hTERT) and human telomerase-associated protein(hTEP1) was determined by RT-PCR assay. Expression of Bax and Bcl-2 proteins was assessed by Western blotting. Results: Celecoxib can inhibit proliferation and induce apoptosis in a dose- and time-dependent manner, repress telomerase activity, decrease hTERT mRNA and Bcl-2 protein expression and increase Bax protein expression, PGE2 had no effect on telomerase. Conclusions: Celecoxib had the antiproliferative and pro-apoptotic effect in Hep-2 cells. Apoptosis was accompanied by a decrease in telomerase activity which was directly correlated with hTERT mRNA and up-regulation of Bax/Bcl-2. Bcl-2 may thus play an important role in telomerase activity as well as apoptosis.

• 한국콘텐츠학회:학술대회논문집
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• 한국콘텐츠학회 2009년도 춘계 종합학술대회 논문집
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• pp.415-419
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• 2009

본 연구는 운동선수의 자기관리가 정신력에 미치는 영향을 규명하기 위하여 2009년 D시에 소재하고 있는 체육계열 중고등학생과 대학교 운동선수를 모집단으로 선정, 최종 353명을 자료분석에 활용하였다. 본 연구에서 사용된 조사도구는 설문지였으며, 설문문항에 대한 신뢰도 검증결과 각 변인 하위척도의 신뢰도의 경우 .720에서 .898로 나타나 비교적 신뢰할만한 수준인 것으로 나타났으며. 타당도 분석결과 KMO 지수가 자기관리 .925, 정신력 .892로 나타나 요인분석을 위한 변수선정이 적절하였음이 밝혀졌으며, Bartlett 구형검정치의 경우 자기관리 및 정신력 등 모든 변인에서 통계적으로 유의미하게 요인분석이 적합한 것으로 나타났다. 본 연구에서 사용된 통계기법은 상관분석(correlation), 회귀분석(regression)이었다. 이와 같은 연구방법을 통해 얻어진 결과를 종합하여 도출시킨 결론은 다음과 같다. 첫째, 자기관리는 자기효능감에 영향을 미친다. 둘째, 자기관리는 걱정에 부분적으로 영향을 미친다. 셋째, 자기관리는 도전의식에 영향을 미친다.

• 폴리머
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• 제25권6호
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• pp.893-901
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• 2001

조직공학적 신경재생 및 파킨슨씨병 등의 시경퇴행성 질환에서의 치료에 이용 목적으로 신경성장인자(nerve growth factor, NGF)를 생분해성 고분자 담체에 NGF를 서방화시키고자 PLA 담체에 함유시켜 유화동결건조법으로 제조하였다. 제조된 NGF의 방출량은 생체외 pH 7.4, 37$^{\circ}C$의 PBS 조건하에서 4주 동안 방출실험 하였으며, 함유된 NGF의 활성을 확인하기 위하여 PC-l2 세포에 직접 배양하여 확인하였다. 제조되어진 PLA 담체는 열린 셀 구조를 가졌으며, 초기 NGF의 함량이 많을수록 방출량도 증가를 보였으며, 제조과정에서의 NGF의 환성을 확인하기 위하여 PC-12 세포를 배양한 결과 신경돌기가 성장하였다. 본 연구는 생분해성 고분자 특정인 확산과 분해에 의해서 생물학적 활성물질인 NGF의 방출을 조절할 수 있으며, 조직공학적으로 서방화되어 3차원적인 신경재생을 가능케 할 것으로 기대된다.

• 한방안이비인후피부과학회지
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• 제22권2호
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• pp.60-73
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• 2009

Objective : Houttuyniae Herba is widely used in oriental medicine as a remedy for inflammation. However, as yet there is no clear explanation of how Houttuyniae Herba affects the production of inflammatory cytokine. This study was to determine the effects of Essence extracted from Houttuynia cordata Thunb(HCT) on the mast cell-mediated inflammatory responses. Method : We measured the amount of inflammatory cytokine production induced by the phorbol myristate acetate (PMA) plus calcium ionophore(A23187) in the human mast cell line (HMC-1) incubated with various concentrations of HCT. The TNF-$\alpha$, IL-6 and IL-8 secreted protein levels were measured by the ELISA assay. The TNF-$\alpha$, IL-6 and IL-8 mRNA levels were measured by the RT-PCR analysis. Nuclear and cytoplasmic proteins were exmined by Western blot analysis. The NF-$\kappa$B promoter activity was examined by a luciferase assay. Result : HCT inhibited the PMA + A23187-induced TNF-$\alpha$, IL-6 expression and reduced mRNA of TNF-$\alpha$, IL-6 and IL-8. we observed that HCT suppressed the induction of NF-B activity. In addition, HCT suppressed PMA plus A23187-induced NF-$\kappa$B promoting activity. Conclusion : In this study, we have found that HCT is an inhibitor of NF-$\kappa$B and cytokines on the mast cell-mediated inflammatory responses.

• 한국동물위생학회지
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• 제32권4호
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• pp.335-341
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• 2009

Canine brucellosis produce abortions and infertility in dogs and is currently diagnosed by serological methods such as rapid slide agglutination test with 2-mercaptoethanol (2-ME RSAT) and immunochromatographic assay (ICA). Bacterial isolation is considered gold standard for Brucella diagnosis and the polymerase chain reaction (PCR) is an alternative method to bacterial isolation. A total of 36 whole blood samples were collected from dogs reared in area of Chuncheon and were subjected to serology (2-ME RSAT and ICA for B. canis, Rose Bengal test and C-ELISA for B. abortus), blood culture and 3 types of PCRs (BSCP31, 16s rRNA, and OMP-2). All blood samples were negative by serology and blood cultures. The BCSP31 and the OMP-2 PCR detected 5 samples were positive whereas the 16S rRNA PCR detected all samples were negative as serological methods and blood culture did. From the results observed in the present study, we conclude that 16S rRNA PCR could be used for direct PCR for canine blood samples.

• 대한수의학회지
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• 제48권3호
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• pp.267-274
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• 2008

This study was to evaluate the efficacy of single dose Mycoplasma hyopneumoniae (M. hyo)-vaccination in the swine farms which had different serological patterns of porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2). A minimum of 240 pigs from each farm was applied, allocating M. hyo vaccinated and control groups. The PRRSV and PCV2 infections were analyzed by serological method (commercial ELISA kit). After administrating pigs a single dose of M. hyo vaccine or control saline at 3 weeks of age, serum antibodies to M. hyo, PRRSV and PCV2 were monitored at 4, 10, 16 and 22 weeks of age. Mortality, weight changes, feed conversion ratio (FCR) and lung score were also evaluated. A single-dose vaccination of M. hyo bacterin was efficacious to reduce mycoplasmal lung lesions and induce good humoral immune response. However, FCR was improved only in one of the three farms where showed seronegative status to both PRRSV and PCV2 in the period from 4 to 16 weeks of age. These results might imply that M. hyo vaccine alone could not overcome the PRRSV and PCV2 infection-associated wasting in the field condition. Therefore, the control of PRRSV and PCV2 should be considered to obtain the better effects of M. hyo vaccination.

• 대한본초학회지
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• 제27권4호
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• pp.9-16
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• 2012

Objective : Canavalia gladiata DC semen (CGS) have been used to improve hematopoietic activity. In the current study, we investigated whether CGS regulate hemato-potentiating function using hematopoietic stem cells (HSCs) as a testing system. Methods : HSCs isolated from femur in mice with leukopenia and thrombocytopenia induced induced by CTX. Then, Real-time PCR was performed to measure the mRNA expression and hematopoietic related gene (EPO, IL-3, SCF, c-kit, GM-CSF), the phoaphorylation of GATA-1 and STAT-5a/b were observed by ELISA method, and the number of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E), semisolid clonogenic assay was performed. Result : When HSCs were treated with CGS, the expression of hematopoietic related genes (EPO, IL-3, SCF, c-kit, and GM-CSF) were significantly increased at the levels of mRNA as well as production in HSCs. Additionally, CGS enhanced phosphorylation of STAT-1 and signal transducer and activator of transcription-5a/b (STAT-5a/b) in HSCs. Furthermore, CGS significantly enhanced the growth rate of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E) in vitro. Conclusion : These result suggest that CGS has hematopoietic enhancement via hematopoietic cytokine-mediated GATA-1/STAT-5a/b pathway.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제35권1호
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• pp.7-12
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• 2009

Purpose: Gelatin-hydroxyapatite nanocomposite is similar to inorganic nanostructure of bone. To make a scaffold with osteoinductivity, bone marrow derived stem cells from rabbit femur were impinged into the nanocomposite. This vitro study was to test osteogenic differentiation of the stem cells in the nanocomposite, which was made by authors. Material & Methods: Gel-HA nanocomposite with 10g of HA, 3 g of Gel has been made by co-precipitation process. Bone marrow was obtained from femur of New Zealand White rabbits and osteogenic differentiation was induced by culturing of the BMSCs in an osteogenic medium. The BMSCs were seeded into the Gel-HA nanocomposite scaffold using a stirring seeding method. The scaffolds with the cells were examined by scanning electron microscopy (SEM), colorimetry assay, biochemical assay with alkaline phosphatase (ALP) diagnostic kit, osteocalcin ELISA kit. Results: Gel-HA nanocomposite scaffolds were fabricated with relatively homogenous microscale pores ($20-40{\mu}m$). The BMSCs were obtained from bone marrow of rabbit femurs and confirmed with flow cytometry, Alizarin red staining. Attachment and proliferation of BMSCs in Gel-HA nanocomposite scaffold could be identified by SEM, ALP activity and osteocalcin content of BMSCs. Conclusion: The Gel-HA nanocomposite scaffold with micropores could be fabricated and could support BMSCs seeding, osteogenic differentiation.