Eukaryotic gene expression is an important process, which is initiated by several transcription factors and RNA polymerases that occupy the promoter region of genomic DNA. Although there are many experiments to identify the promoter region in a gene, it is time and labor consuming to finalize it. In this study, we utilized bioinformatic programs, including Ensembl, NCBI, and CpG plots, to identify the cloning promoter region in SETDB1 genomic DNA. We performed PCR amplification to obtain the SETDB1 promoter on an approximately 2 kb region upstream from the TSS named SETDB1-P1. The PCR product was ligated with TA cloning vectors, and we confirmed the insert size using restriction enzyme digestion. Sequentially, the insert was subcloned into a pGL3-luc vector to produce pGL3-SETDB1- P1-luc and then confirmed by DNA sequencing. We also obtained a fragmented PCR product called P2 and P3 and performed a luciferase assay using pGL3-SETDB1-P1-luc transfection. We found that several anticancer drugs, including taxol, 4-FU, and doxorubicin, decreased the promoter activity of SETDB1. We obtained consistent data on the regulation of SETDB1 gene expression after anticancer drug treatment using Western blot analysis and RT-PCR. Our results suggest that promoter cloning of the human SETDB1 gene utilizing bioinformatics is a very useful and timesaving approach to study gene expression.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
/
v.39
no.6
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pp.274-282
/
2013
Objectives: The posterior maxillary region often provides a limited bone volume for dental implants. Maxillary sinus elevation via inserting a bone graft through a window opened in the lateral sinus wall has become the most common surgical procedure for increasing the alveolar bone height in place of dental implants in the posterior maxillary region. The purpose of this article is to assess the change of bone volume and the clinical effects of dental implant placement in sites with maxillary sinus floor elevation and autogenous bone graft through the lateral window approach. Materials and Methods: In this article, the analysis data were collected from 64 dental implants that were placed in 24 patients with 29 lacks of the bone volume posterior maxillary region from June 2004 to April 2011, at the Department of Oral and Maxillofacial Surgery, Inha University Hospital. Panoramic views were taken before the surgery, after the surgery, 6 months after the surgery, and at the time of the final follow-up. The influence of the factors on the grafted bone material resorption rate was evaluated according to the patient characteristics (age and gender), graft material, implant installation stage, implant size, implant placement region, local infection, surgical complication, and residual alveolar bone height. Results: The bone graft resorption rate of male patients at the final follow-up was significantly higher than the rate of female patients. The single autogenous bone-grafted site was significantly more resorbed than the autogenous bone combined with the Bio-Oss grafted site. The implant installation stage and residual alveolar height showed a significant correlation with the resorption rate of maxillary sinus bone graft material. The success rate and survival rate of the implant were 92.2% and 100%, respectively. Conclusion: Maxillary sinus elevation procedure with autogenous bone graft or autogenous bone in combination with Bio-Oss is a predictable treatment method for implant rehabilitation.
Journal of the korean academy of Pediatric Dentistry
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v.42
no.3
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pp.218-225
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2015
This study used sodium fluorescein to improve imaging diagnostic ability by increasing the fluorescence difference between sound enamel and caries lesions. It also made it easier to discriminate between stain and caries lesions using quantitative light-induced fluorescence (QLF). Half of the specimen surface was covered with nail varnish as a control. Specimens were divided randomly in six decalcification groups and decalcified for different lengths of time. Then, ${\Delta}F$ was measured using QLF-D. After applying 0.075% sodium fluorescein, we measured ${\Delta}F$ again and compared it with the initial value. After cutting the central portion of the specimen, we measured the lesion depth using scanning electron microscopy. The lesion surfaces observed with QLF were darker than normal enamel, whereas they were lighter than normal enamel after applying fluorescein. Longer decalcification time was associated with greater fluorescent dye penetration. The ${\Delta}F$ measured after applying fluorescein was higher than the initial value (p < 0.05). Due to QLF measurement using fluorescein being more sensitive for diagnosing early decalcification, this approach will enable early diagnosis of dental caries before the cavity formation stage, allowing the treatment of early caries lesions. With QLF and sodium fluorescein, we can easily discriminate between stain and caries lesions.
Journal of the korean academy of Pediatric Dentistry
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v.40
no.1
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pp.60-65
/
2013
The term 'triple tooth' is used to describe a rare dental abnormality in which three teeth appear to be joined. The literature contains many reports of joined primary teeth; most cases have involved, however, the joining of two teeth, and only rarely three teeth. Triple tooth has clinical problems such as dental caries, esthetic problems, malocclusions, and periodontal problems. Therefore, it may require multi-disciplinary approach. The present study describes rare case of triple tooth between maxillary primary central and lateral incisors and a supernumerary tooth. An one-year, eleven-month old girl was seen for evaluation of swelling in the labial gingiva above a right maxillary triple tooth. She experienced traumatic dental injury in that area three weeks ago. Intraoral examination revealed an abscess and fistula in the region of the triple tooth. A radiographic examination showed that right lateral incisor was missing. Endodontic treatment and composite resin restoration was performed on the triple tooth. After follow-ups of 7 months period, there were no marked complications.
Pulmonary sequestration is a very rare congenital malformation in which a mass of pulmonary tissue is detached from the normal lung and receives its blood supply from a systemic artery. It may be clinically asymptomatic or it has a wide spectrum of various clinical manifestations. The clinical therapeutic approach is to resect the sequestered lobe to prevent frequent complication such as infection. The arterial embolization of feeding artery is a new technique and a less invasive treatment than conventional surgical removal. We have experienced a 17-year-old male with pulmonary sequestration whose complaints were pain in left lower chest. He was diagnosed by computed tomography and aortography and successfully treated with embolization of feeding artery. We report a case of pulmonary sequestration treated with arterial embolization instead of surgery.
Zizyphus jujube leaf fractions (ZLFs) showed no cytotoxic effects of up to $100{\mu}g/mL$, while the anti-inflammatory effects of ZLFs were analyzed by checking the productions of nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$), cyclooxygenase-2 (COX-2), and inflammatory cytokines TNF-${\alpha}$, IL-$1{\beta}$, and IL-6 in the lipopolysaccharide (LPS)-stimulated Raw264.7 macrophage up to the concentration of $100{\mu}g/mL$. ZLFs ($100{\mu}g/mL$) demonstrated a strong anti-inflammatory activity that reduced 61~85% of NO and 71~100% of $PGE_2$ production in the LPS-stimulated Raw264.7 macrophage. Even the low ZLFs concentration of $1{\mu}g/mL$ have reduced NO and $PGE_2$ production by 34~64%. Expressions of COX-2 protein were also effectively inhibited by the ZLFs. Furthermore, the TNF-${\alpha}$, IL-$1{\beta}$, and IL-6 production were significantly suppressed through the treatment of ZLFs at concentrations of 1, 10, and $100{\mu}g/mL$. In the order of the Zizyphus jujube leaf water fraction (ZLWF) < buthanol fraction (ZLBF) < ethyl acetate fraction (ZLEF) showed anti-inflammatory activity. In particular, the ethyl acetate fraction ZLEF at $100{\mu}g/mL$ showed an excellent anti-inflammatory activity by reducing the production of NO, $PGE_2$, COX-2, and inflammatory cytokines (TNF-${\alpha}$, IL-$1{\beta}$, IL-6) in the level of Raw264.7 macrophage without LPS-stimulation or even better. The results of our study suggest the potential of ZLEF for use as an excellent ant-inflammatory inhibiting mediator and may be used as a therapeutic approach to various inflammatory diseases.
This study investigated relative explanation of social support, ego-resiliency and stress coping strategies to help adaptive coping style of school-dropout adolescents under stress situation. To this end, 101 school-dropout adolescents were surveyed by visiting and requesting the outofschool youth supporting project, youth detention center, and adolescent protective and treatment facilities in Daejeon, Cheongnam, and Chungbuk. As analysis methods, descriptive statistical analysis, pearson's correlation, and hierarchical analysis were conducted and the research results are as follows. First, stress coping strategies showed positive relationship with social support and ego-resiliency. Second, a variable that showed greater explanation power for stress coping strategies was the environmental variable, the social support. Third, it was identified that there was greater explanation power when the environmental variable, the social support, and the personal variable, the ego-resiliency, were put in at the same time for stress coping strategies. According to the result, this study implies that schools, community, national policy effort and systemetic approach are required as well as improvement of personal coping capabilities in order to overcome difficulties school-dropout adolescents face.
Purpose: Unresectable rectal cancer has a grave prognosis. regardless of the therapy used and median survival is less than 1 rear. Also, it is reported by many authors that $50-80\%$ of unresectable lesions were rendered resectable by radiation therapy and the median survival time for the completely resected patients were better than that of the unresected patients. So we analyzed retrospectively our data for the better treatment outcome in these patients. Materials and MEthods:From 1980 to 1992, 45 patients with initially unresectable tumors in the rectum were treated with radiation therapy with/without surgery in Department of Radiation Oncology, Yonsei Cancer Center 10 MV radiation and multiple field technique (box or AP/PA) were used. The total dose was 28-70 Gy and median dose was 48 Gy. We evaluated the lesion status at 45-50 Gy for operability. If the lesions appeared to be resectable, the Patients were operated on 4-6 weeks after radiation therapy. But if the lesions were still fixed, the radiation dose was increased to 60-65 Gy. Results: For all patients, the 2-year actuarial survival was $13.3\%$ and median survival was 9.5 months. Of 6 patients who had received less than 45 Gy, only $17\%$ of patients responded, but in the patients who had received more than 45 Gy, $60\%$ of response rate was achieved Six of the 24 patients$(25\%)$ underwent surgical resections following RT. For patients undergoing curative resection. the two-rear survival was $50\%,$ but that of the patients without resection was $9.5\%$ (p<0.01). Survival of patients with complete response following RT was $50\%$ at 2 years. Survival of patients with partial response, stable disease and progressive disease after RT was $13.4\%,\;15.4\%,\;0\%$ respectively (P<0.05). Conclision: Our data suffests that the efforts which can increase the response rate and aggressive surgical approach are needed to achieve the better local control and survival in unresectable rectal cancers.
Kim Kye-Seong;Lim Jung-Jin;Yang Yun-Hee;Kim Soo-Kyoung;Yoon Tae-Ki;Cha Kwang-Yul;Lee Dong-Ryul
Journal of Microbiology and Biotechnology
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v.16
no.9
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pp.1347-1354
/
2006
The aims of this study were to establish a simple and effective method for isolating male germline stem cells (GSCs), and to test the possibility of using these cells as a new approach for male infertility treatment. Testes obtained from neonatal and adult mice were manually decapsulated. GSCs were collected from seminiferous tubules by a two-step enzyme digestion method and plated on gelatin-coated dishes. Over 5-7 days of culture, GSCs obtained from neonates and adults gave rise to large multicellular colonies that were subsequently grown for 10 passages. During in vitro proliferation, oct-4 and two immunological markers (Integrin ${\beta}1,\;{\alpha}6$) for GSCs were highly expressed in the cell colonies. During another culture period of 6 weeks to differentiate to later stage germ cells, the expression of oct-4 mRNA decreased in GSCs and Sertoli cells encapsulated with calcium alginate, but the expression of c-kit and testis-specific histone protein 2B(TH2B) mRNA as well as the localization of c-kit protein was increased. Expression of transition protein (TP-l) and localization of peanut agglutinin were not seen until 3 weeks after culturing, and appeared by 6 weeks of culture. The putative spermatids derived from GSCs supported embryonic development up to the blastocyst stage with normal chromosomal ploidy after chemical activation. Thus, GSCs isolated from neonatal and adult mouse testes were able to be maintained and proliferated in our simple culture conditions. These GSCs have the potential to differentiate into haploid germ cells during another long-term culture.
Background: Establishment of laparoscopic cholecystectomy as an outpatient procedure has accentuated the clinical importance of reducing early postoperative pain, as well as postoperative nausea and vomiting (PONV). We therefore planned to evaluate the role of a multimodal approach in attenuating these problems. Methods: One hundred and twenty adult patients of ASA physical status I and II and undergoing elective laparoscopic cholecystectomy were included in this prospective, randomized, placebo-controlled study. Patients were divided into four groups of 30 each to receive methylprednisolone 125 mg intravenously or etoricoxib 120 mg orally or a combination of methylprednisolone 125 mg intravenously and etoricoxib 120 mg orally or a placebo 1 hr prior to surgery. Patients were observed for postoperative pain, fentanyl consumption, PONV, fatigue and sedation, and respiratory depression. Results were analyzed by the ANOVA, a Chi square test, the Mann Whitney U test and by Fisher's exact test. P values of less than 0.05 were considered to be significant. Results: Postoperative pain and fentanyl consumption were significantly reduced by methylprednisolone, etoricoxib and their combination when compared with placebo (P<0.05). The methylprednisolone + etoricoxib combination caused a significant reduction in postoperative pain and fentanyl consumption as compared to methylprednisolone or etoricoxib alone (P<0.05); however, there was no significant difference between the methylprednisolone and etoricoxib groups (P>0.05). The methylprednisolone and methylprednisolone + etoricoxib combination significantly reduced the incidence and severity of PONV and fatigue as well as the total number of patients requiring an antiemetic treatment compared to the placebo and etoricoxib (P<0.05). Conclusions: A preoperative single-dose administration of a combination of methylprednisolone and etoricoxib reduces postoperative pain along with fentanyl consumption, PONV, antiemetic requirements and fatigue more effectively than methylprednisolone or etoricoxib alone or a placebo.
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