• 한국수정란이식학회지
• /
• 제31권1호
• /
• pp.1-7
• /
• 2016

Xenotransplantation involves multiple steps of immune rejection. The present study was designed to produce nuclear transfer embryos, prior to the production of transgenic pigs, using fibroblasts carrying transgenes human complement regulatory protein hCD59 and interleukin-18 binding protein (hIL-18BP) to reduce hyperacute rejection (HAR) and cellular rejection in pig-to-human xenotransplantation. In addition to the hCD59-mediated reduction of HAR, hIL-18BP may prevent cellular rejection by inhibiting the activation of natural killer cells, activated T-cell proliferation, and induction of $IFN-{\gamma}$. Transgene construct including hCD59 and ILI-18BP was introduced into miniature pig fetal fibroblasts. After antibiotic selection of double transgenic fibroblasts, integration of the transgene was screened by PCR, and the transgene expression was confirmed by RT-PCR. Treatment of human serum did not affect the survival of double-transgenic fibroblasts, whereas the treatment significantly reduced the survival of non-transgenic fibroblasts (p<0.01), suggesting alleviation of HAR. Among 337 reconstituted oocytes produced by nuclear transfer using the double transgenic fibroblasts, 28 (15.3%) developed to the blastocyst stage. Analysis of individual embryos indicated that 53.6% (15/28) of embryos contained the transgene. The result of the present study demonstrates the resistance of hCD59 and IL-18BP double-transgenic fibroblasts against HAR, and the usefulness of the transgenic approach may be predicted by RT-PCR and cytolytic assessment prior to actual production of transgenic pigs. Further study on the transfer of these embryos to surrogates may produce transgenic clone miniature pigs expressing hCD59 and hIL-18BP for xenotransplantation.

• 생명과학회지
• /
• 제23권8호
• /
• pp.989-997
• /
• 2013

금은화의 메탄올 추출에 의한 유기 용매별 분획에서 항산화 효과를 근거로 에틸 아세테이트 분획에서 $H_2O_2$로 유도된 RAW 264.7 세포독성에 대한 세포보호 효과와 유전자 발현 양상을 분석하였다. 용매별 분획의 항산화 활성은 시료의 농도가 증가할수록 DPPH에 대한 전자공여능도 증가하였으며, $ED_{50}$은 에틸 아세테이트 분획에서 $39.56{\mu}g/ml$로 가장 높게 나타났다. 또한 $H_2O_2$에 의해 유도된 Raw 264.7 세포사멸(($IC_{50}$)에 대하여 에틸 아세테이트 분획은 농도 의존적으로 유의적인 세포 생존율과, $100{\mu}g/ml$의 농도에서 82.49%의 세포보호 효과를 나타내었다. 한편 $IC_{50}$의 $H_2O_2$ 독성에 대한 에틸 아세테이트 분획의 Raw 264.7 세포보호 효과에 관련된 유전자 발현의 패턴은 BHA와 유사한 것으로 나타났다. 항산화 관련 유전자군의 비교 분석에서는 에틸 아세테이트 분획과 BHA는 매우 유사한 유전자 발현 양상을 보였으나, 세포 내 주요 항산화 효소를 만드는 유전자 중 하나인 heme oxygenase 1 (Hmox1) 유전자의 발현 정도가 에틸 아세테이트 분획에서 4 배 이상 증가하였다. 항염증 관련 유전자군의 비교에서는 T-box transcription factor (Tbx21)가 BHA 처리군에서는 2 배 감소하였으나 에틸 아세테이트 분획의 처리군에서는 오히려 약 2 배 증가한 것으로 나타났다.

• 사상체질의학회지
• /
• 제15권2호
• /
• pp.137-150
• /
• 2003

The purpose of this study is to examine the toxic effects caused by glucose oxidase (GO) and the effects of Hyungbangjihwangtang (HJT) water extracts on the treatment of the toxic effects. For this purpose, experiments with the cultured hippocampal cells from new born mice were done. The results of these experiments were as follows. 1. GO decreased the survival rate of the cultured cells on MTT assay and NR assay. 2. HJT has the efficacy of decreasing the lipid peroxidation. 3. HJT has the efficacy of increasing the amount of neurofilaments. 4. HJT has the efficacy of decreasing the activation of protein kinase C(PKC). From the above results, it is concluded that HJT has marked efficacy as a treatment for the damages caused by the GO-mediated oxidative stress. Further clinical study of this pharmacological effects of H]T should be completed.

• Journal of Microbiology and Biotechnology
• /
• 제28권1호
• /
• pp.157-164
• /
• 2018

Francisella tularensis (FT), a highly infectious pathogen, is considered to be a potential biological weapon owing to the current lack of a human vaccine against it. Tul4 and FopA, both outer membrane proteins of FT, play an important role in the bacterium's immunogenicity. In the present study, we evaluated the immune response of mice - humanized with human CD34+ cells (hu-mice) - to a cocktail of recombinant Tul4 and FopA (rTul4 and rFopA), which were codon-optimized and expressed in Escherichia coli. Not only did the cocktail-immunized hu-mice produce a significant human immunoglobulin response, they also exhibited prolonged survival against an attenuated live vaccine strain as well as human T cells in the spleen. These results suggest that the cocktail of rTul4 and rFopA had successfully induced an immune response in the hu-mice, demonstrating the potential of this mouse model for use in the evaluation of FT vaccine candidates.

• Asian Pacific Journal of Cancer Prevention
• /
• 제15권21호
• /
• pp.9459-9465
• /
• 2014

Background: Structural maintenance of chromosomes 4 (SMC-4) is a chromosomal ATPase which plays an important role in regulate chromosome assembly and segregation. However, the role of SMC-4 in the incidence of malignancies, especially colorectal cancer is still poorly understood. Materials and Methods: We here used quantitative PCR and Western blot analysis to examine SMC-4 mRNA and protein levels in primary colorectal cancer and paired normal colonic mucosa. SMC-4 clinicopathological significance was assessed by immunohistochemical staining in a tissue microarray (TMA) in which 118 cases of primary colorectal cancer were paired with noncancerous tissue. The biological function of SMC-4 knockdown was measured by CCK8 and plate colony formation assays. Fluorescence detection has been used to detect cell cycling and apoptosis. Results: SMC-4 expression was significantly higher in colorectal cancer and associated with T stage, N stage, AJCC stage and differentiation. Knockdown of SMC-4 expression significantly suppressed the proliferation of cancer cells and degraded its malignant degree. Conclusions: Our clinical and experimental data suggest that SMC-4 may contribute to the progression of colorectal carcinogenesis. Our study provides a new therapeutic target for colorectal cancer treatment.

• Asian Pacific Journal of Cancer Prevention
• /
• 제17권8호
• /
• pp.4043-4048
• /
• 2016

Background: Head and neck cancer (HNC) is the ninth most common cancer worldwide, and has a poor 5-year survival rate averaging 50%, which has not changed for decades. A high prevalence of HNC has been reported in the southwestern region of Saudi Arabia, as compared to other areas of the country. However, data in regards to HNC are scattered and not well documented. Thus, the aim of this systematic review was to gather all available and updated important information regarding HNC in Saudi Arabia, and highlight the gaps of knowledge in our country with regard to this disease. In addition, suggestions of solutions to overcome the current status and improve our future standard of care to fight HNC are also highlighted. Materials and Methods: The electronic databases PubMed and Google Scholar using English-language literature were used for this systematic review, using specific inclusion and exclusion criteria and keywords. The search was performed in April 2016 and updated in June 2016. Results: Our search revealed twenty-one studies that fulfilled our inclusion and exclusion criteria and that were conducted in Saudi Arabia. These studies investigated different aspects of HNC, including prevalence, risk factors, biomarkers, and assessed knowledge and awareness of both public and practitioners with regard to HNC. Conclusions: This review uncovered a big gap in our epidemiological data in cancer information in general, and head and neck cancer in particular. In addition, a lack of knowledge and awareness of both the public and health care practitioners hinders the early diagnosis of disease and negatively impact the prognosis, treatment and outcome. The Ministry of Health in Saudi Arabia should develop a more systematic way and adapt policies to gather cancer information in general, and head and neck cancer in particular, from all governmental and private sectors from all over the kingdom, and develop educational programs to raise the knowledge and awareness of HNC in the country.

• 대한방사선방어학회:학술대회논문집
• /
• 대한방사선방어학회 2011년도 추계 학술발표회 및 심포지엄
• /
• pp.212-213
• /
• 2011

The biological effects of low dose ionizing radiation (LDIR) remain insufficiently understood. We examined for the scientific evidence to show the biological effects of LDIR using radiation-sensitive immune cells. We found that Ikaros protein was responsed to low dose-dependent effects of gamma radiation in IM-9 B lymphocytes. Ikaros encodes zinc finger transcription factors that is important regulators of a hematopoietic stem cells (HSCs) progression to the B lymphoid lineage development, differentiation and proliferation. In this study, we observed that cell proliferation was enhanced from 10% to 20% by LDIR (0.05 Gy) in IM-9 B lymphocytes. The Ikaros protein was phosphorylated in its serine/threonine (S/T) region and decreased its DNA binding activity in the cells exposed to LDIR. We found that Ikaros phosphorylation was up-regulated by CK2/AKT pathway and the residues of ser-304 and ser-306 in Ikaros was phosphorylated by LDIR. We also observed that Ikaros protein was localized from the nucleus to the cytoplasm after LDIR and bound with Autotaxin (ENPP2, ATX) protein, stimulating proliferation, migration and survival of immune cells. In addition, we found that the lysoPLD activity of ATX was dependent on Ikaros-ATX binding activity. These results indicate that the Ikaros is an important regulator of immune activation. Therefore, we suggest that low dose ionizing radiation can be considered as a beneficial effects, stimulating the activation of immune cells.

• 한국발생생물학회지:발생과생식
• /
• 제16권2호
• /
• pp.129-135
• /
• 2012

선행연구에서 본 연구자는 IEX-1 단백질이 난소 암세포에서 세포사멸(apoptosis)을 유도하는 기능을 수행함을 확인하였으나, 세포사멸과 세포생존의 여러 단계에서 어떠한 신호전달 체계로 IEX-1이 작용하는지는 정확히 알지 못하고 있다. 따라서 IEX-1 단백질과 결합하는 새로운 단백질을 찾기 위해 yeast two-hybrid system을 이용하였다. 그 결과 IEX-1이 여러 다양한 인간 암 세포에서 세포사멸을 유도하는 CATHEPSIN B 단백질과 결합함을 밝혔다. 본 연구에서는 리소좀 프로테아제의 일원인 CATHEPSIN B 단백질과 IEX-1 단백질의 결합을 면역침강법과 western blot 분석으로 확인하였다. 그러므로 이 결과들을 통해서 IEX-1과 CATHEPSIN B는 세포 내에서 서로의 기능에 영향을 미칠 것으로 예상되며, 세포사멸을 유도하는 IEX-1 단백질이 lysosome-mediated apoptotic pathway에 관여할 가능성을 시사한다.

• Asian Pacific Journal of Cancer Prevention
• /
• 제14권2호
• /
• pp.987-991
• /
• 2013

Aim: To study anti-tumor effects of exosomes from class II transactivator (CIITA) gene transfected CT26 cells. Methods: In this study, we established an MHC class II molecule-expressing murine colon cancer cell line (CT26-CIITA) by transduction of the CIITA gene. Immune effects in vitro and tumor protective results in vivo were tested and monitored. Results: Exosomes from CT26-CIITA cells were found to contain a high level of MHC class II protein. When loaded on dendritic cells (DCs), exosomes from CT26-CIITA cells significantly increased expression of MHC class II molecules, CD86 and CD80, as compared to exosomes from CT26 cells. In vitro assays using co-culture of immunized splenocytes and exosome-loaded DCs demonstrated that CIITA-Exo enhanced splenocyte proliferation and IFN-${\gamma}$ production of CD4+T cells, while inhibiting IL-10 secretion. In addition, compared to exosomes from CT26 cells, CT26-CIITA-derived exosomes induced higher TNF-${\alpha}$ and IL-12 mRNA levels. A mouse tumour preventive model showed that CT26-CIITA derived exosomes significantly inhibited tumour growth in a dose-dependent manner and significantly prolonged the survival time of tumour-bearing mice. Conclusion: Our findings indicate that CT26-CIITA-released exosomes are more efficient to induce anti-tumour immune responses, suggesting a potential role of MHC class II-containing tumour exosomes as cancer vaccine candidates.

• 동의생리병리학회지
• /
• 제25권2호
• /
• pp.217-226
• /
• 2011

The aim of this study is to identify central neural pathway of neurons following the projection to the large intestine and Hapgok(LI4) which is Won acupoint of the large intestine meridian of hand-yangmyeong. In this experiment, Bartha's strain of pseudorabies virus was used to trace central localization of neurons related with large intestine and acupoint(LI4) which has been known to be able to regulate intestinal function. The animals were divided into 3 groups: group 1, injected into the large intestine; group 2, injected into the acupoint(LI4); group 3, injected into the acupoint(LI4) after severing the radial, ulnar, median nerve. After four days survival of rats, PRV labeled neurons were identified in the spinal cord and brain by immunohistochemical method. First-order PRV labeled neurons following the projection to large intestine, acupoint(LI4) and acupoint(LI4) after cutting nerve were found in the cervical, thoracic, lumbar and sacral spinal cord. Commonly labeled neurons were labeled in the lumbosacral spinal cord and thoracic spinal cord. They were found in lamina V- X, intermediomedial nucleus and dorsal column area. The area of sensory neurons projecting was L5-S2 spinal ganglia and T12-L1 spinal ganglia, respectively. In the brainstem, the neurons were labeled most evidently and consistently in the nucleus tractus solitarius, area postrema, dorsal motor nucleus of vagus nerve, reticular nucleus, raphe nuclei(obscurus, magnus and pallidus), C3 adrenalin cells, parapyramidal area(lateral paragigantocellular nucleus), locus coeruleus, subcoeruleus nucleus, A5 cell group, periaqueductal gray matter. In the diencephalon, PRV labeled neurons were marked mostly in the arcuate nucleus and median eminence. These results suggest that overlapped CNS locations are related with autonomic nuclei which regulate the functions of large intestine-related organs and it was revealed by tracing PRV labeled neurons projecting large intestine and related acupoint(LI4).