• Asian Pacific Journal of Cancer Prevention
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• v.17 no.7
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• pp.3643-3646
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• 2016

Management of eyelid cancers is based on surgery and/or radiotherapy (RT). The treatment objective is to control tumors with acceptable functional and esthetic outcomes. The aim of this study was to evaluate the results of radiation therapy in management of epithelial eyelid cancers, reviewing retrospectively the clinical records of patients treated in our institution from January 1989 to December 2013. We focused on clinical and histological features, treatment characteristics, tolerance and disease control. One hundred and eight patients (62 men and 46 women) were enrolled, with a mean age of 61 years [ranges 15-87]. The most frequent tumor location was the inner canthus (42.6%). Median tumor size was 21 mm [ranges 4-70]. Histological type was basal cell carcinoma in 88 cases (81.5%), squamous cell carcinoma in 16 (14.8%) and sebaceous carcinoma in 4 (3.7%). Radiation therapy was exclusive in 67 cases (62%) and post-operative for positive or close margins in the remaining cases. Kilovoltage external beam radiotherapy (KVRT) was used in 63 patients (58.3%) and low-dose-rate interstitial brachytherapy in 37 (34.3%). Eight (7.4%) were treated with cobalt or with a combination of KVRT-cobalt, KVRT-electron beams, KVRT-brachytherapy or cobalt-electron beams. The total delivered radiation doses were 70 Gy (2 Gy/fraction) in 62 patients (57.4%), 66 Gy (2 Gy/fraction) in 37 (34.3%) and 61.2 Gy (3.4Gy/fraction) in 9 (8.3%). After a median follow-up of 64 months, we noted 10 cases of local recurrences(9.2%): 7 after exclusive and 3 after post-operative RT. No local recurrence occurred in patients treated with brachytherapy. Actuarial 5-year local recurrence-free rate, disease-free survival and overall survival were respectively 90%, 90% and 97%. T-stage was found to be a significant factor for recurrence (p=0.047). All acute radiation-related reactions were scored grade I or II. Delayed effects were eye watering in 24 cases (22.2%), eye dryness in 19 (17.6%), unilateral cataract in 7 (6.4%) and ectropion in 4 (3.7%). Radiation therapy and especially brachytherapy is an efficient treatment of eyelid cancers, allowing eye conservation and functional preservation with good local control rates and acceptable toxicity.

• Clinical and Experimental Pediatrics
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• v.56 no.11
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• pp.482-489
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• 2013

Purpose: The aim of the present study was to investigate the differences in lower airway inflammatory immune responses, including cellular responses and responses in terms of inflammatory mediators in bronchoalveolar lavage fluid (BALF) and the airway, to rhinovirus (RV) infection on asthma exacerbation by comparing a control and a murine asthma model, with or without RV infection. Methods: BALB/c mice were intraperitoneally injected with a crude extract of Dermatophagoides farinae (Df ) or phosphate buffered saline (PBS) and were subsequently intranasally treated with a crude extract of Df or PBS. Airway responsiveness and cell infiltration, differential cell counts in BALF, and cytokine and chemokine concentrations in BALF were measured 24 hours after intranasal RV1B infection. Results: RV infection increased the enhanced pause (Penh) in both the Df sensitized and challenged mice (Df mice) and PBS-treated mice (PBS mice) (P<0.05). Airway eosinophil infiltration increased in Df mice after RV infection (P<0.05). The levels of interleukin (IL) 13, tumor necrosis factor alpha, and regulated on activation, normal T cells expressed and secreted (RANTES) increased in response to RV infection in Df mice, but not in PBS mice (P<0.05). The level of IL-10 significantly decreased following RV infection in Df mice (P<0.05). Conclusion: Our findings suggest that the augmented induction of proinflammatory cytokines, Th2 cytokines, and chemokines that mediate an eosinophil response and the decreased induction of regulatory cytokines after RV infection may be important manifestations leading to airway inflammation with eosinophil infiltration and changes in airway responsiveness in the asthma model.

• Journal of Periodontal and Implant Science
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• v.35 no.3
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• pp.549-561
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• 2005

이 연구의 목적은 새로이 제작된 chitosan-poly(L-lactic acid)(PLLA) 다층 다공성 차폐막의 생체적합성 및 골세포활성도 및 골재생능을 평가하는 것이다. 제작된 차폐막을 24 well에 넣고 clonal osteoblast-like cell line(MC3T3-E1)을 접종한 군을 실험군으로, 차폐막을 사용하지 않은 대조군으로 하였다. 배양 1일, 7일 및 14일째에 각 well에서 세포수를 측정하였다. 주사전자현미경을 이용하여 차폐막에 부착된 세포의 형태관찰을 시행하였다. RNA 추출 및 RT-PCR을 실시한 후, agarose gel상에서 전기영동하여 조골세포 표식자인 collagen type I(COL), osteopontin(OP) 및 osteocalcin(OC) mRNA의 발현을 관찰하였다. 제작된 매트릭스의 생체적합성 및 골재생능을 관찰하기 위하여 백서의 두개골에 직경 8mm의 원형 결손부를 형성한 후 차폐막을 이식한 군을 실험군으로, 아무 것도 넣지 않은 군을 대조군으로 하여 4주 경과 후 실험동물을 희생시킨 후 조직학적관찰을 시행하였다. 시간경과에 따른 부착세포수 관찰결과, 배양 14일까지 조골세포의 수가 지속적으로 증가하였고, 주사전자현미경으로 세포의 형태 관찰결과, 배양된 세포들은 중층의 형태로 성장하면서 시간경과에 따라 세포가 응집되는 양상을 나타내었다. 관찰 기간동안 COL, OP, 및 OC mRNA의 발현이 관찰되어 배양 전 기간동안 조골세포의 형질이 잘 유지되고 있음을 알 수 있었다. 백서 두개골 결손부에 이식된 차폐막은 염증반응 없이 주위 조직과 우수한 생체적합성을 나타내었으며, 차폐막을 이식하지 하지 않은 대조군에 비해 높은 신생골 형성을 나타내었다. 이상의 관찰결과로 새로이 제작된 chitosan-PLLA 차폐막은 우수한 생체적합성 및 골재생능을 나타냄을 알 수 있었으며, 향후 이를 골조직 재생 및 치주조직유도재생 분야에 응용될 수 있을 것으로 생각된다.

• Clinical and Experimental Reproductive Medicine
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• v.24 no.3
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• pp.399-405
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• 1997

Progesterone is necessary for successful pregnancy and had immunosuppressive properties. Peripheral blood mononuclear cells (PBMC) from many women with unexplained recurrent spontaneous abortion responded to trophoblast extract in vitro by prolifertion and releasing soluble, heat-labile factors that are toxic to mouse embryos (embryotoxic factors). Accumulating evidence suggests that T Helper (Th)-1 type immunity to trophoblast is correlated with embryotoxic factor production and is associated with pregnancy loss, while Th2-type immunity is associated with successful gestation. The objective of this study was to determine whether progesterone can inhibit Th1-type cytokine secretion (IFN-${\gamma}$, TNF-${\alpha}$) by trophoblast-activated peripheral blood mononuclear cells from 23 nonpregnant women (age 25-35) with unexplained recurrent abortion (median 5, range 3 to 15)who otherwise produce embryotoxic factors in response to trophoblast. We also determined whether progesterone affected Th2-type cytokines (IL-4, IL-10) in this system in vitro and if IL-10 (1,500 pg/mL) could inhibit Th1-type immunity to trophoblast. IFN-${\gamma}$ was detected in 17 of 23 (74%) trophoblast stimulated PBMC culture supernatants ($77.94{\pm}23.79$ pg/mL) containing embryotoxic activity. TNF-${\alpha}$ was detected in 19 (83%) of these same supernatants ($703.15{\pm}131.36$ pg/mL). In contrast, none of the supernatants contained detectable levels of IL-4 or IL-10. Progesterone ($10^{-5}$, $10^{-7}$, $10^{-9}$M) inhibited Th1-type immunity in a dose dependent manner, but had no effect on Th2-type cytokine secretion. The inhibitory effects of progesterone were abrogated with RU486, but did not affect Th2-type cytokine secretion in trophoblast-activated cell cultures. IL-10, like progesterone also inhibited Th1-type cytokine secretion but had no effect on Th2-type cytokines. These data suggest that therapies designed to suppress Th1-type cytokine secretion in women with recurrent abortion who have evidence of Th1-type immunity to trophoblast may be efficacious in preventing pregnancy loss and should be tested in appropriately designed clinical trials.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.51-51
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• 2001

Cocaine and amphetamine-regulated transcript (CART) is a satiety factor that is regulated by leptin. It was reported that the mice intracerebroventricularly injected with CART showed behavioral changes resembled with the typical behavioral alterations found in the mice carrying disorders in the brain serotonergic (5-HT) system. Hence, this study was conducted to find out the relationships between CART and 5-HT. We first examined the mRNA levels of CART after the injections of para-chlorophenylalanine (pCPA, 300 mg/kg i.p., single injection or daily for three consecutive days) in the rat brains by in situ hybridization using the mouse CART cDNA probe cloned in our laboratory. Systemic administrations of pCPA, a potent inhibitor of tryptophan hydroxylase, the rate limiting enzyme of 5-HT biosynthesis, acutely depletes the brain 5-HT transporter (5-HTT) in the dorsal raphe nucleus (DRN), which reuptakes terminal 5-HT. Results indicated that the mRNA level of CART significantly decreased in the arcuate nucleus, paraventricular nucleus, and lateral hypothalamic nucleus by three days of daily injection with pCPA with no noticeable change detected 24 hrs after the single injection. The message levels of 5-HTT in DRN decreased in both single and three days of injections. Secondly, to investigate whether CART affect to 5-HT, mouse genomic CART gene, which is consist of 3 exons and 2 introns and mouse neurofilament light (NF-L) chain promoter were cloned. Then, we constructed neuron specific expression vector, which was transfected into HeLa cell using lipid-mediated transfection system. Expression of GFP and CART linked to NF-L-chain promoter in the transfected HeLa cell were detected by using fluorescent microscope and RT-PCR. These results confirmed normal expression of DNA constructs in vitro. Then, to increase brain specific expression of CART in vivo transgenic mice carrying CART gene controlled the deleted NF-L-chain promoter were generated by the DNA microinjection into pronuclei of fertilized embryos. Transgenic mice were detected by Southern blot. Further study is necessary to examine CART expression and 5-HTT in these transgenic mice. Therefore, these results suggest that there maybe a positive molecular correlation between CART and 5-HT in responding to the stimuli.

• Journal of Korean Society of Forest Science
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• v.97 no.3
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• pp.215-220
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• 2008

Recently, there has been a great deal of interest in the applications of plant-based extracts to both cosmetic and medicinal industries. The objective of this study was to investigate the anti-inflammatory and antimicrobial effect of P. rigida extracts by water and ethyl acetate. Anti-inflammatory and anti-microbial effect of P. rigida extracts by water and EtOAc were investigated by using nitrite scavenging ability, nitric oxide production and anti-microbial ability. In the test of nitrite scavenging ability, P. rigida extracts by water and EtOAc showed 88.7% and 99% at 100 ppm concentration, respectively. The cell viability was measured using the MTT assay at 24 hours after P. rigida extracts as shown in over 80%. Anti-inflammatory effect was examined in LPS stimulated RAW 264.7 cells. NO productions in LPS and P. rigida extracts stimulated group were decreased in a concentration and were dependent on time as compared with LPS stimulated. The water extracts showed the highest inhibition at the 100 ppm concentration. In anti-microbial activity test, the water extract with 3.0 mg/disc resulted in the clear zone of 14 mm, and ethyl acetate with that of 15 mm for Staphylococcus aureus. However, P. rigida extracts didn't show any growth inhibitory effect on Esherichia coli. These results indicate that the extracts of P. rigida have anti-inflammatory activities as a cosmeceuticals.

• Journal of Periodontal and Implant Science
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• v.30 no.2
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• pp.213-231
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• 2000

이 연구의 목적은 다공성의 CPP 내부에 쥐의 장골의 골수에서 유래된 세포를 접종하고 3차 원적으로 배양하여 CPP가 골 형성을 위한 조직공학의 지지체로 적용가능한가를 연구하는 것과 Calcium PolyPhosphate(CPP)의 돌연변이 유발성을 검사하는 것이다. 무수 ($Ca(H_2PO_4)$)를 condensation하여 무결정의 ($Ca(PO_3)$)를 얻고 이를 용융하고 냉각시킨 후 분쇄하여 Calcium polyphosphate(CPP) powder를 얻었다. 다공성의 CPP는 5% $SiO_2$를 첨가하여 sponge 형태로 $450-550{\mu}m$ 소공의 크기를 가지는 것과(CPP-45ppi) $200-300{\mu}m$의 소공의 크기를 가지는 것(CCP-60ppi) 2가지로 제작하였다. 각각의 CPP matrices는 $5mm{\times}5mm{\times}1mm$의 블록 형태로 만들었다. 체중 100g 내외의 백서에서 장골(femur, tibia)을 채취하여 백서의 장골 골수 세포를 분리하여 배양한 후 24well에 CPP block을 넣고 CPP block 당 $10^5$개의 배양한 세포를 접종하였다. 배양 1, 7, 14, 및 21 일째에 각 well에서 trypsin EDTA를 이용하여 2회 반복하여 cell을 분리하였고, 원심분리한 후 hemacytometer로 측정하였다. 또, 45ppi와 60ppi, 그 리 고 Tissue Culture Polystyrene(control group)에 접종, 배양된 세포들의 염기성 인산분해효소활성도를 배양 7, 14, 및 21 일째에 각각 측정하였다. 각 기간별로 배양된 세포-CPP 혼합체내에서 세포의 부착 및 증식과 형성된 조직의 3차원적 형태를 관찰하기 위하여 주사전자현미경하에서의 관찰하였다. CPP의 돌연변이 유발성 검사 (mutagenicity test)를 위해 hypoxanthine-guanine phosphoribosyl transferase(HPRT) assay를 하였다. NIH3T3 cell line과 CHO-K1 cell line으로 각각 $1000{\mu}g/m{\ell}$, $100{\mu}g/m{\ell}$, $10{\mu}g/m{\ell}$ 그리고 $1{\mu}g/m{\ell}$의 CPP 농도에서 측정하였다. 통계적 분석을 위해서 모든 측정은 각군당 4개체 이상 시험하였고, 각 측정값은 평균값${\pm}$표준편차로 나타내었다. 각 군간의 통계적 유의성 검정을 위해서 Analysis of variance(ANOVA)를 이용하였고 Tukey의 방법으로 사후분석을 실시하였다. 제작된 CPP matrices 소공들이 서로간에 연결이 잘 되어있는 형태였다. 두 가지로 제조된 CPP(45ppi와 60ppi) 모두에서 세포의 부착이 잘 일어났고, 부착된 세포의 분열도 잘 일어났다. 2 가지의 CPP 모두에서 7, 14, 21일째의 세포 수는 1일째에 비해 유의성 있게 증가하였다(P<0.01). 3차원적 구조인 Calcium PolyPhosphate에서 배양한 세포는 24well dish(tissue culture polystyrene)에서 평면적으로 배양한 대조군의 세포에서 보다 염기성 인산분해효소 (Alkaline Phosphatase)를 유의성 있게 높게 나타냈다. 주사전자현미경에서 세포-CPP 혼합체를 관찰한 결과, CPP block에 세포들이 잘부착되어 있었고, 시간이 지남에 따라 세포가 여러 층을 형성하면서 뭉치는 현상을 보였다. 또, HPRT assay 결과 , Calcium PolyPhosphate는 돌연변이 유발성을 보이지 않았다. 이상의 결과로 볼 때 CPP에는 세포부착이 잘 일어나고, 지지체 상에서 세포의 분열도 활발하게 일어나므로 골조직을 위한 조직공학의 우수한 지지체가 될 수 있을 것으로 사료된다.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.28 no.5
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• pp.375-395
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• 2006

Purpose: Lingual nerve (LN) damage may be caused by either tumor resection or injury such as wisdom tooth extraction, Although autologous nerve graft is sometimes used to repair the damaged nerve, it has the disadvantage of necessity of another operation for nerve harvesting. Moreover, the results of nerve grafting is not satisfactory. The nerve growth factor (NGF) is well-known to play a critical role in peripheral nerve regeneration and its local delivery to the injured nerve has been continuously tried to enhance nerve regeneration. However, its application has limitations like repeated administration due to short half life of 30 minutes and an in vivo delivery model must allow for direct and local delivery. The aim of this study was to construct a well-functioning $rhNGF-{\beta}$ adenovirus for the ultimate development of improved method to promote peripheral nerve regeneration with enhanced and extended secretion of hNGF from the injured nerve by injecting $rhNGF-{\beta}$ gene directly into crush-injured LN in rat model. Materials and Methods: $hNGF-{\beta}$ gene was prepared from fetal brain cDNA library and cloned into E1/E3 deleted adenoviral vector which contains green fluorescence protein (GFP) gene as a reporter. After large scale production and purification of $rhNGF-{\beta}$ adenovirus, transfection efficiency and its expression at various cells (primary cultured Schwann cells, HEK293 cells, Schwann cell lines, NIH3T3 and CRH cells) were evaluated by fluorescent microscopy, RT-PCR, ELISA, immunocytochemistry. Furthermore, the function of rhNGF-beta, which was secreted from various cells infected with $rhNGF-{\beta}$ adenovirus, was evaluated using neuritogenesis of PC-12 cells. For in vivo evaluation of efficacy of $rhNGF-{\beta}$ adenovirus, the LNs of 8-week old rats were exposed and crush-injured with a small hemostat for 10 seconds. After the injury, $rhNGF-{\beta}$ adenovirus($2{\mu}l,\;1.5{\times}10^{11}pfu$) or saline was administered into the crushed site in the experimental (n=24) and the control group (n=24), respectively. Sham operation of another group of rats (n=9) was performed without administration of either saline or adenovirus. The taste recovery and the change of fungiform papilla were studied at 1, 2, 3 and 4 weeks. Each of the 6 animals was tested with different solutions (0.1M NaCl, 0.1M sucrose, 0.01M QHCl, or 0.01M HCl) by two-bottle test paradigm and the number of papilla was counted using SEM picture of tongue dorsum. LN was explored at the same interval as taste study and evaluated electro-physiologically (peak voltage and nerve conduction velocity) and histomorphometrically (axon count, myelin thickness). Results: The recombinant adenovirus vector carrying $rhNGF-{\beta}$ was constructed and confirmed by restriction endonuclease analysis and DNA sequence analysis. GFP expression was observed in 90% of $rhNGF-{\beta}$ adenovirus infected cells compared with uninfected cells. Total mRNA isolated from $rhNGF-{\beta}$ adenovirus infected cells showed strong RT-PCR band, however uninfected or LacZ recombinant adenovirus infected cells did not. NGF quantification by ELISA showed a maximal release of $18865.4{\pm}310.9pg/ml$ NGF at the 4th day and stably continued till 14 days by $rhNGF-{\beta}$ adenovirus infected Schwann cells. PC-12 cells exposed to media with $rhNGF-{\beta}$ adenovirus infected Schwann cell revealed at the same level of neurite-extension as the commercial NGF did. $rhNGF-{\beta}$ adenovirus injected experimental groups in comparison to the control group exhibited different taste preference ratio. Salty, sweet and sour taste preference ratio were significantly different after 2 weeks from the beginning of the experiment, which were similar to the sham group, but not to the control group.

• Tuberculosis and Respiratory Diseases
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• v.39 no.4
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• pp.334-342
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• 1992

Background: The activated T lymphocyte by inhalaed mycobacterial antigen may evoke cell-mediated immunity in patients with active pulmonary tuberculosis. These activated lymphocyte may influence the response of tuberculin-purified protein derivative (PPD) in skin test. But occasionally, anergy to PPD appear in patients with pulmonary tuberculosis in spite of active stage. Thus we evaluated the effect of change of subtypes of lymphocyte in bronchoalveolar lavage fluid (BAL) and peripheral blood on anergy to PPD in patients with active pulmonary tuberculosis. Method: We performed tuberculin skin test and flow-cytometry analysis of lymphocytes obtained from BAL fluid and peripheral blood in 11 healthy normal volunteers and 20 patients with active pulmonary tuberculosis. Results: 1) The composition of lymphocyte significantly increased in patients with active pulmonary tuberculosis when compared with that in healthy control ($25.2{\pm}4.8$ vs $6.5{\pm}1.3%$, p<0.01), but composition of monocyte significantly decreased ($69.6{\pm}5.7$ vs $89.2{\pm}1.4%$, p<0.05) in analysis of BAL fluid. 2) There were no differences in compositions of cells in BAL fluid between responders and no-responders to PPD. 3) The compositions of CD3 (+), CD4 (+), CD3 (+) IL-2R (+), CD3 (+) HLA-DR (+) significantly increased in BAL fluid when compared with those in peripheral blood in patients with active pulmonary tuberculosis. But the composition of CDS (+), CD4/CDS were not different between BAL fluid and peripheral blood. 4) There were no correlations between response to PPD and compositions of cells and lymphocyte subtypes in BAL fluid and peripheral blood in all patients with tuberculosis, responders, and no-responders, respectively. Conclusion: From these results, we suggest no direct relationship between compositions of inflammatory cells in bronchoalveolar lavage fluid and we could not rule out the possibility of compartmentalization of activated lymphocyte involving in anergy to PPD in skin test in patients with active pulmonary tuberculosis.

• Tuberculosis and Respiratory Diseases
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• v.46 no.1
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• pp.44-52
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• 1999

Background : Airway infiltration by inflammatory cells, particularly of eosinophils, is one of the characteristic features of asthma. Several mechanisms for the recruitment of eosinophil is focused on the CD4+ T lymphocyte for the preferential production of Th2-c1erived cytokines. Interleukin-10(IL-10) is identified cytokine with potent antiinflammatory activity. This molecule has been shown to inhibit the release of cytokine from inflammatory cells including Th2 cell, and also to inhibit eosinophil survival. We therefore attempted to determine whether decreased synthesis of IL-10 in the lung of bronchial asthma may contribute to inflammation that is characteristics of this dease. Method: Subjects were patients with bronchial asthma(n=23) and normal controls(n=11). IL-10 produced from peripheral mononuclear cell(PBMC) and in bronchoalveolar lavage(BAL) fluid was measured by ELISA method. Degree of bronchial inflammation was assessed by total cell counts and eosinophil percents in BAL fluid, eosinophil infiltration on bronchial biopsy tissue and $PC_{20}$ for methacholine. Results: The IL-10 level produced by PBMC and in BAL fluid from patient with bronchial asthma were not different with normal controls(respectively, $901.6\pm220.4$ pg/ml, $810.9\pm290.8$ pg/ml for PBMC, $24.5\pm9.5$ pg/mL $30.5\pm13.5$ pg/ml for BAL fluid p>0.05). There were significant negative correlation between IL-10 in BAL fluid and eosinophil percents in BAL fluid or degree of eosinophil infiltration in bronchial biopsy (respectively r=-0.522, r=-0.4486 p<0.05). However there was no difference of IL-10 level according to $PC_{20}$ for methacholine. There were no correlation between IL-10 production by PBMC and peripheral blood eosinophil counts or serum eosinophilic cationic protein levels(respectively r=0.1146, r=0.0769 p>0.05). Conclusion: These observation suggest that IL-10 may participate but not acts the crucial role in regulation of the airway inflammation in bronchial asthma.