• Title/Summary/Keyword: 조직염색법

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The Correlations of Parameters Using Contrast Enhanced Ultrasonography in the Evaluation of Prostate Cancer Angiogenesis (전립선암쥐모형의 신생혈관생성의 평가를 위해 시행된 역동적 조영 증강 초음파에서 얻은 변수간의 상관성연구)

  • Hwang, Sung Il;Lee, Hak Jong;Kim, Kil Joong;Chung, Jin-haeng;Jung, Hyun Sook;Jeon, Jong June
    • Ultrasonography
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    • v.32 no.2
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    • pp.132-142
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    • 2013
  • Purpose: The purpose of this study is to investigate the correlations of various kinetic parameters derived from the time intensity curve in a xenograft mouse model injected with a prostate cancer model (PC-3 and LNCaP) using an ultrasound contrast agent with histopathologic parameters. Materials and Methods: Twenty nude mice were injected with human prostate cancer cells (15 PC-3 and five LNCaP) on their hind limbs. A bolus of $500{\mu}L$ ($1{\times}10^8$ microbubbles) of second-generation US contrast agent (SonoVue) was injected into the retroorbital vein. The region of interest was drawn over the entire tumor. The time intensity curve was acquired and then fitted to a gamma variate function. The maximal intensity (A), time to peak (Tp), maximal wash-in rate (washin), washout rate (washout), area under the curve up to 50 sec ($AUC_{50}$), area under the ascending slope ($AUC_{in}$), and area under the descending slope ($AUC_{out}$) were derived from the parameters of the gamma variate fit. Immunohistochemical staining for VEGF and CD31 was performed. Tumor volume, the area percentage of VEGF stained in a field, and the count of CD31 (microvessel density, MVD) positive vessels showed correlation with the parameters from the time intensity curve. Results: No significant differences were observed between the kinetic and histopathological parameters from each group. MVD showed positive correlation with A (r=0.625, p=0.003), washin (r=0.462, p=0.040), $AUC_{50}$ (r=0.604, p=0.005), and $AUC_{out}$ (r=0.587, p=0.007). Positive correlations were also observed between tumor volume and $AUC_{50}$ (r=0.481, p=0.032), washin (r=0.662, p=0.001), and $AUC_{out}$ (r=0.547, p=0.012). Washout showed negative correlations with MVD (r=-0.454, p=0.044) and tumor volume (r=-0.464, p=0.039). The area percentage of VEGF did not show any correlation with calculated data from the curve. Conclusion: MVD showed correlations with several of the kinetic parameters. CEUS has the potential for prediction of tumor vascularity in a prostate cancer animal model.

Gastroduodenoscopic Findings and Effect of Therapy of Helicobacter pylori Infection in Children (소아 Helicobacter pylori 감염의 상부 위장관 내시경 소견 및 치료 효과)

  • Rhee, Kyung Shin;Park, Jae Ock
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.8 no.1
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    • pp.12-20
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    • 2005
  • Purpose: Helicobacter pylori infection is known to be associated with acute or chronic abdominal pain and upper gastrointestinal bleeding in children. This study was performed to analyze the gastroduodenoscopic findings and the efficacy of triple therapy with omeprazole, amoxicillin and clarithromycin between one and two weeks of duration in children with H. pylori infection. Methods: We have assessed retrospectively 60 patients presented with acute or chronic abdominal pain or upper gastrointestinal bleeding. H. pylori infection was confirmed by endoscopic biopsy and rapid urease test. Out of 60 patients, 30 patients were treated with a combination of omeprazole, amoxicillin, and clarithromycin for one week, and the other 30 patients were treated for two weeks with the same medication. Efficacy of treatment was assessed 4 weeks after the termination of treatment by using the $^{13}C$ urea breath test. Results: The 60 patients with the complaint of diffuse abdominal pain, epigastric pain, vomiting or hematemesis were included in this study. One-week treatment group (group I) consisted of 30 patients (14 male, 16 female) with mean age of $11.6{\pm}2.67years$. Two-week treatment group (group II) consisted of 30 patients (11 male, 19 female) with mean age of $10.7{\pm}4.17years$. In group I, H pylori were eradicated in 26 out of 30 patients (86.7%). In group II, H. pylori were eradicated in 26 out of 30 children (86.7%). Both groups did $^{13}C$ urea breath test after 4 weeks after termination of the triple therapy. The eradication rates were same in both groups as 86.7%, 26 out of 30 patients in each group. The results of endoscopy were nodular gastritis 26 (43.3%), erosive gastritis 10 (16.7%), hemorrhagic gastritis 7 (11.7%), gastric ulcer 2 (3.3%) and normal finding 15 (25.0%). Conclusion: In this study, the nodular gastritis was most common endoscopic findings with H. pylori positive patients. The eradication rate of H. pylori with omeprazole, amoxicillin and clarithromycin was 86.7% and it would be highly effective as primary treatment with no significant differences in the eradication rate between one-week and two-week treatment groups. However, we should need more long-term follow-up data.

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Prognostic Value of the Expression of p53 and bcl-2 in Non-Small Cell Lung Cancer (비소세포폐암에서 p53과 bcl-2의 발현이 예후에 미치는 영향)

  • Yang, Seok-Chul;Yoon, Ho-Joo;Shin, Dong-Ho;Park, Sung-Soo;Lee, Jung-Hee;Keum, Joo-Seob;Kong, Gu;Lee, Jung-Dal
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.5
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    • pp.962-974
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    • 1998
  • Background: Alteration of p53 tumor suppressor genes is most frequently identified in human neoplasms, including lung carcinoma. It is well known that bcl-2 oncoprotein protects cells from apoptosis. Recent studies have demonstrated that bcl-2 expression is associated with favorable prognosis for patients with non-small cell lung carcinoma. However, the precise biologic role of bcl-2 in the development of these tumors is still obscure. p53 and bcl-2 have important regulatory influence in the apoptotic pathway and thus their relationship is of interest in tumorigenesis, especially lung cancer. Purpose: The author investigated to know the prognostic significance of the expression of p53 and bcl-2 in radically resected non-small cell lung cancer. Method: 84 cases of formalin-fixed paraffin-embedded blocks from resected primary non-small cell lung cancer from 1980 to 1994 at Hanyang University Hospital were available for both clinical follow-up and immunohistochemical staining using monoclonal antibodies for p53 and bcl-2. Results : The histologic classification of the tumor was based on WHO criteria., and the specimens included 45 squamous cell carcinomas(53.6%), 28 adeonocarcinomas(33.3%) and 11 large cell carcinomas(13.1 %). p53 immunoreactivity was noted in 47 cases of 84 cases(56.0%). bcl-2 immunoreactivity was noted in 15 cases of 84 cases(17.9%). The mean survival duration was $64.23{\pm}10.73$ months in bcl-2 positive group and $35.28{\pm}4$. 39 months in bcl-2 negative group. The bcl-2 expression was significantly correlated with survival in radically resected non-small cell lung cancer patients(p=0.03). The mean survival duration was $34.71{\pm}6.12$ months in p53 positive group and $45.35{\pm}6.30$ months in p53 negative group(p=0.21). The p53 expression was not predictive for survival. There was no correlation between combination of the different status of p53 and bcl-2 expression in our study. Conclusions : The interaction and the regulation of new biologic markers, such as those involved in the apoptotic pathway, are complex. bcl-2 overexpression is a good prognostic factor in non-small cell lung cancer and p53 expression is not significantly associated with the prognostic factor in non-small cell lung cancer.

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Expression of Nesfatin-1/NUCB2 and Its Binding Site in Mouse Ovary (생쥐 난소 내 Nesfatin-1/NUCB2 발현과 결합 부위 확인)

  • Kim, Jin-Hee;Youn, Mi-Ra;Bang, So-Young;Sim, Ji-Yeon;Kang, Hee-Rae;Yang, Hyun-Won
    • Development and Reproduction
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    • v.14 no.4
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    • pp.287-295
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    • 2010
  • It was recently reported that nesfatin-1/NUCB2, which is secreted from the brain, controls appetite and energy metabolism. The purpose of this research was to confirm whether or not the protein and its binding site should have been expressed in the mouse reproductive organs and to know the possible effects of nesfatin-1 on the reproductive function. Using the ICR female mouse ovary and uterus, the expression of NUCB2 mRNA was confirmed with the conventional PCR and the relative amount of NUCB2 mRNA in the tissues was analyzed with real-time PCR. Immunohistochemical staining was performed using the nesfatin-1 antibody to investigate the nesfatin-1 protein expression and the biotin conjugated nesfatin-1 to confirm the binding site for nesfatin-1 in the ovary. Furthermore, in order to examine if the expression of NUCB2 mRNA in the ovary and uterus is affected by gonadotropin, its mRNA expression was analyzed after PMSG administration into mice. As a result, the expression level of NUCB2 mRNA in the ovary and the uterus was as much as the expression level in hypothalamus. As a result of the immunohistochemical staining, nesfatin-1 proteins were localized at the theca cells, the interstitial cells, and some of the luteal cells. However, the granulosa cells in the follicles did not stain. Interestingly, the oocytes in the some follicles were stained with nesfatin-1. On the other hand, nesfatin-1 protein binding sites were displayed at the theca cells and the interstitial cells near the tunica albuginea. After PMSG administration the expression level of NUCB2 mRNA was increased in the ovary and the uterus. These results demonstrate that for the first time the nesfatin-1 and its binding site were expressed in the ovary and NUCB2 mRNA expression was controlled by gonadotropin, suggesting an important role in the reproductive organs as a local regulator. Therefore, further study is needed to elucidate the functions of nesfatin-1 on the reproductive organs.

Gene Expression of Candidate Genes Involved in Fat Metabolism During In vitro Adipogenic Differentiation of Bovine Mesenchymal Stem Cell (Bovine Mesenchymal Stem Cell의 지방분화를 이용한 지방대사관련 후보 유전자의 발현분석)

  • Kim, Sung-Kon;Kim, Nam-Kuk;Yoon, Du-Hak;Kim, Tae-Hun;Yang, Boo-Keun;Lee, Hyun-Jeong
    • Journal of Animal Science and Technology
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    • v.52 no.4
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    • pp.265-270
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    • 2010
  • Adipogenesis has been one of the most intensely studied models of cellular differentiation. During adipogenesis, differential expression of many adipogenesis related genes lead to profound changes in cellular, morphological, and physiological characteristics of the differentiating cells. The aim of the present study was to examine the expression levels of adipogenic candidate genes, cAMP early repressor (ICER), nephroblastoma over-expressed protein (NOV), heat shock protein beta 1 (HSPB1) and succinate dehydrogenase (SDH), during adipogenesis of bovine mesenchymal stem cells (BMSC). The BMSC were cultured in DMEM / low glucose medium with adipogenic inducers for 6 days and the expression of various candidate genes which seemed related to adipogenesis were measured by real-time PCR. This study showed that the expression of peroxisome proliferator activated receptor ${\gamma}$(PPAR${\gamma}$) and fatty acid binding protein 4 (FABP4) genes as adipogenic indicators were increased to 3.11 and 3.11 folds on day 6 than on day 0, respectively (p<0.05). To determine whether candidate genes were related to adipogenesis, the expression levels of ICER, NOV, HSPB1, and SDH genes were measured during adipogenesis in BMSC. Our results showed that the expression level of ICER gene was significantly increased to 4.12 folds (0.01729 vs. 0.07138; p<0.05), whereas NOV, HSPB1, and SDH genes were decreased to 2.89, 3.18 and 2.36 folds, respectively, on day 6 when compared to day 0. These results suggest that these candidate genes have stimulatory or inhibitory effects on adipogenesis in BMSC, indicating that these genes may be directly or indirectly related to the adipogenic event of adipose precursor cells.

Relationship Between Vascularity and Other Remodeling Parameters in Asthmatic Airway (기관지천식환자에서 기도내 혈관분포정도(vascularity) 및 다른 기도 재구성 소견과의 연관성)

  • Kim, Seung Joon;Lee, Sook Young;Kim, Myoung Sook;Lo, Dae Keun;Kwon, Soon Seog;Kim, Young Kyoon;Kim, Kwan Hyoung;Moon, Hwa Sik;Song, Jeong Sup;Park, Sung Hak
    • Tuberculosis and Respiratory Diseases
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    • v.54 no.2
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    • pp.191-198
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    • 2003
  • Background : The pathological features in asthmatic airway remodeling are diverse. The aim of this study was to examine the degree of airway vascularity in relation to the other remodeling parameters in asthmatics. Methods : Bronchial biopsies were done in 34 asthmatic patients, and 6 control subjects. The basement membrane thickness and the subepithelial thickness were measured in the hematoxylin-eosin stained tissue, and the degree of vascularity was measured using type IV collagen immunostaining. Results : 1) Compared to the control subjects, the asthmatics showed a significant increase in the basement membrane thickness ($6.92{\pm}2.01{\mu}m$ vs $9.67{\pm}2.84{\mu}m$, p<0.05) and the subepithelial thickness ($44.49{\pm}31.92{\mu}m$ vs $121.22{\pm}72.79{\mu}m$, p<0.05). 2) Compared to the control subjects, the asthmatics showed a significant increase in the vascular area per unit submucosal area ($4.51{\pm}2.13%$ vs $10.32{\pm}6.08%$, p<0.05). In addition, the number of vessels per unit submucosal area showed an increased tendency without statistical significance. 3) In the asthmatics, the number of vessels and the vascular area per unit submucosal area showed no correlation with the basement membrane thickness, the subepithelial thickness, the severity, the forced expiratory volume in 1 second($FEV_1$), and the methacholine provocative concentration 20($PC_{20}$). Conclusion : This study showed that vascularity was an important parameter in asthmatic airway remodeling but it was not related to the other remodeling parameters such as the basement membrane thickness and the subepithelial thickness. Each of these asthmatic remodeling parameters may have a different clinical significance. Therefore, further studies will be needed.

Helicobacter pylori reinfection rate by a 13C-urea breath test and endoscopic biopsy tests in Korean children (한국 소아에서 Helicobacter pylori 박멸 후 13C-요소 호기 검사와 내시경적 생검을 이용한 재감염률 연구)

  • Shim, Jeong Ok;Seo, Jeong Kee
    • Clinical and Experimental Pediatrics
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    • v.49 no.3
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    • pp.268-272
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    • 2006
  • Purpose : The reinfection rate of H. pylori reported before $^{13}C$-urea breath test($^{13}C$-UBT) era was higher than that of the post $^{13}C$-UBT era. Children are usually reluctant to receive invasive endoscopic evaluation for the reinfection of H. pylori, particularly when they are asymptomatic. The aim of the study is to discover the reinfection rate by different diagnostic tests, and to find out what causes the difference. Methods : Children confirmed to be eradicated from H. pylori were included in the study. Reinfection was evaluated by endoscopic biopsy based tests(n=34, mean age $11.5{\pm}3.7$ years) and/or a $^{13}C$-UBT(n=38, mean age $10.0{\pm}3.6$ years) at the time of 18 months after eradication. At first visit, H. pylori infection had been diagnosed by positive results from a rapid urease test, Giemsa stain and Warthin-Starry stain and/or a positive culture. Eradication was defined as negative results from all above tests 1-3 months after eradication therapy. Results : Reinfection rate by endoscopic biopsy based tests was 35.3 percent(12/34). All patients had abdominal symptoms(P=0.000). Reinfection rate was 13.2 percent(5/38) by a $^{13}C$-UBT. Reinfection rate was higher in children with abdominal symptoms(P=0.008). There was no evidence that reinfection rate depended on the sex(P=0.694), age(P=0.827), diseases(peptic ulcers vs gastritis, P=0.730) and eradication regimen(P=0.087). Conclusion : Helocibacter pylori reinfection rate in Korean children was 13.2 percent per 18 months by a non-invasive test or $^{13}C$-UBT. Accurate determinations of the reinfection rate in children is affected by the compliance of the diagnostic tests. Non-invasive tests should be considered to investigate the reinfection rate in children.

Preparation of Active Fraction from Radish Water Extracts for Improving the Intestinal Functions and Constipation Activities (무(Raphanuse sativa var. nigra L.) 물 추출물로 부터 장기능 및 변비질환 개선을 위한 활성 분획의 제조)

  • Baik, Soon-Ok;Lee, Yoo-Hui;Kim, Young-Sook;Ryu, Myeong-Hyeon;Kim, Hyun-Kyung
    • Applied Biological Chemistry
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    • v.47 no.3
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    • pp.315-320
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    • 2004
  • The aim of this study was to develop an activator, 3-10 kDa fraction from radish water extracts, that will improve the intestinal function and bowel movement in the colons. Radish water extracts were investigated for their intestinal function effects according to the charcoal meal transit method, employing Balb/c mice: also, their anti-constipation activities were compared utilizing the loperamide-induced constipation method, employing SD rats. The result suggested that the effects of the charcoal meal transit increased remarkably in radish water extract administrated rats in comparison to loperamide administrated rats. Futhermore, the effects of various solvent extracts of radish on charcoal meal transit in Balb/c mice increased remarkably in radish water fraction administrated rats than in different solvent fraction administrated rats. Radish extraction was tested and isolated into 4 groups: below 3 kDa, 3-10 kDa, 10-300 kDa, and over 300 kDa. 3-10 kDa was the most effective on the intestinal function and bowel movement in the colons; also, 3-10 kDa fraction of radish water extraction was found to be the most effective charcoal meal transit. The dry weight and moisture content of feces remarkedly increased in the 3-10 kDa administrated rats group than in the loperamide only group. Experimental results revealed that 3-10 kDa fraction of radish water extract was the most effective on the intestinal function and bowel movement was the crypt epithelial cells that contained more MUC2 in the 3-10 kDa administrated group than the loperamide only group: in addition, the thickness of mucus layer stained with alcian blue was significantly thicker in 3-10 kDa administrated rats than in loperamide administrated rats. Crypt epithelial cells secreted more MUC2 in the 3-10 kDa administrated group than the loperamide only group and the stained cells clearly showed the MUC2 with antibody Biogenex AM358.

Comparison of TheraCal LC, Mineral trioxide aggregate, and Formocresolas pulpotomy agents in rat molar (백서에서 치수절단술에 사용하는 TheraCal LC, MTA 그리고 Formocresol의 비교)

  • Lee, Bin-Na;Song, Young-Sang;Lee, Go-Woon;Kim, Young-Hoon;Chang, Hoon-Sang;Hwang, Yun-Chan;Oh, Won-Mann;Hwang, In-Nam
    • Korean Journal of Dental Materials
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    • v.44 no.2
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    • pp.187-195
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    • 2017
  • TheraCal LC, a new light-cured, resin-modified calcium silicate-filled base/liner material, has been introduced as a pulpotomy agent. The aim of this study was to evaluate the capacity of hard tissue formation and pulpal response after pulpotomy with TheraCal LC. Twenty-two 9-week-old male rats were anesthetized, cavities were prepared in maxillary first molars and pulps were capped with formocresol (FC), mineral trioxide aggregate (MTA), and TheraCal LC. Specimens obtained from rats were scanned using a high-resolution micro CT system. The specimens were prepared and evaluated histologically, and immunofluorescence assay was performed to assess the dentin matrix protein-1 (DMP-1) expression. On micro CT analysis, the MTA and TheraCal LC groups showed thicker hard tissue formation than the FC group. On hematoxylin and eosin (H&E) staining, MTA and TheraCal LC groups showed dentine bridge formation with vital pulp beneath the materials. On immunofluorescence analysis, DMP-1 was highly expressed in the TheraCal LC group compared to the FC group. TheraCal LC showed similar capacity to form hard tissue as MTA when it was used as a pulpotomy agent. Because of its good manipulation and faster setting time compared to MTA, TheraCal LC could be considered as a good alternative to MTA.

In Vitro Differentiation of Human Amniotic Membrane-derived Stem Cells into Hepatocyte-like Cells (양막 유래 줄기세포의 간세포로의 분화 유도)

  • Kook, Min-Ji;Park, Soo-Yeon;Kang, Hyun-Mi;Kim, Hae-Kwon
    • Development and Reproduction
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    • v.10 no.1
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    • pp.63-73
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    • 2006
  • This study aimed to find out suitable culture conditions for the differentiation of human amniotic membrane-derived stem cells(HAM) into hepatocyte-like cells. Almost homogenous population of fibroblast-like cells was successfully isolated from the amniotic membrane. In comparison to the non-coated plates and in the absence of insulin/transferrin/selenium(ITS), HAM cultured on the fibronectin-coated plates and in the presence of ITS showed the more intense immunocytochemical staining against the albumin. Addition of both fibroblast growth factor(FGF)-1 and -2 to the differentiation medium gave stronger staining compared to the treatment with FGF-1 or -2 alone. Periodic acid Schiff's base staining of glycogen and morphological turnover of fibroblast-like appearance of HAM into round shape matched the results of immunocytochemical studies. When the efficiency of two-step culture method was examined on the differentiation of HAM into hepatocyte-like cells, all of the results of immunocytochemical staining, periodic acid Schiff's staining and morphological change exhibited effective hepatic differentiation of HAM compared to the continuous culture method. Immunoblot analyses of HAM- conditioned media against the albumin showed that the culture of HAM in the presence of both ITS and fibronectin always gave a stronger staining intensity than those in the absence of them, and that the addition of ether mixture of FGF-4 and either FGF-2 or transforming growth $factor(TGF)-{\alpha}$ to the culture medium significantly enhanced the albumin secretion by HAM. Based on these observations, it is suggested that HAM could differentiate into hepatocyte-like cells under a culture condition consisting of fibronectin and ITS, and addition of FGF-4 with either one of FGF-2 or $TGF-{\alpha}$ could enhance the hepatic differentiation of HAM.

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