• Title/Summary/Keyword: methods of data analysis

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Processed Panax ginseng, sun ginseng, inhibits the differentiation and proliferation of 3T3-L1 preadipocytes and fat accumulation in Caenorhabditis elegans

  • Lee, Hyejin;Kim, Jinhee;Park, Jun Yeon;Kang, Ki Sung;Park, Joeng Hill;Hwang, Gwi Seo
    • Journal of Ginseng Research
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    • v.41 no.3
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    • pp.257-267
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    • 2017
  • Background: Heat-processed ginseng, sun ginseng (SG), has been reported to have improved therapeutic properties compared with raw forms, such as increased antidiabetic, anti-inflammatory, and antihyperglycemic effects. The aim of this study was to investigate the antiobesity effects of SG through the suppression of cell differentiation and proliferation of mouse 3T3-L1 preadipocyte cells and the lipid accumulation in Caenorhabditis elegans. Methods: To investigate the effect of SG on adipocyte differentiation, levels of stained intracellular lipid droplets were quantified by measuring the oil red O signal in the lipid extracts of cells on differentiation Day 7. To study the effect of SG on fat accumulation in C. elegans, L4 stage worms were cultured on an Escherichia coli OP50 diet supplemented with $10{\mu}g/mL$ of SG, followed by Nile red staining. To determine the effect of SG on gene expression of lipid and glucose metabolism-regulation molecules, messenger RNA (mRNA) levels of genes were analyzed by real-time reverse transcription-polymerase chain reaction analysis. In addition, the phosphorylation of Akt was examined by Western blotting. Results: SG suppressed the differentiation of 3T3-L1 cells stimulated by a mixture of 3-isobutyl-1-methylxanthine, dexamethasone, and insulin (MDI), and inhibited the proliferation of adipocytes during differentiation. Treatment of C. elegans with SG showed reductions in lipid accumulation by Nile red staining, thus directly demonstrating an antiobesity effect for SG. Furthermore, SG treatment down-regulated mRNA and protein expression levels of peroxisome proliferator-activated receptor subtype ${\gamma}$ ($PPAR{\gamma}$) and CCAAT/enhancer-binding protein-alpha ($C/EBP{\alpha}$) and decreased the mRNA level of sterol regulatory element-binding protein 1c in MDI-treated adipocytes in a dose-dependent manner. In differentiated 3T3-L1 cells, mRNA expression levels of lipid metabolism-regulating factors, such as amplifying mouse fatty acid-binding protein 2, leptin, lipoprotein lipase, fatty acid transporter protein 1, fatty acid synthase, and 3-hydroxy-3-methylglutaryl coenzyme A reductase, were increased, whereas that of the lipolytic enzyme carnitine palmitoyltransferase-1 was decreased. Our data demonstrate that SG inversely regulated the expression of these genes in differentiated adipocytes. SG induced increases in the mRNA expression of glycolytic enzymes such as glucokinase and pyruvate kinase, and a decrease in the mRNA level of the glycogenic enzyme phosphoenol pyruvate carboxylase. In addition, mRNA levels of the glucose transporters GLUT1, GLUT4, and insulin receptor substrate-1 were elevated by MDI stimulation, whereas SG dose-dependently inhibited the expression of these genes in differentiated adipocytes. SG also inhibited the phosphorylation of Akt (Ser473) at an early phase of MDI stimulation. Intracellular nitric oxide (NO) production and endothelial nitric oxide synthase mRNA levels were markedly decreased by MDI stimulation and recovered by SG treatment of adipocytes. Conclusion: Our results suggest that SG effectively inhibits adipocyte proliferation and differentiation through the downregulation of $PPAR{\gamma}$ and $C/EBP{\alpha}$, by suppressing Akt (Ser473) phosphorylation and enhancing NO production. These results provide strong evidence to support the development of SG for antiobesity treatment.

Korean Children and Adolescents with Crohn's Disease Are More Likely to Present with Perianal Fistulizing Disease at Diagnosis Compared to Their European Counterparts

  • Kang, Ben;Kim, Jung Eun;Jung, Jae Hun;Choe, Jae Young;Kim, Mi Jin;Choe, Yon Ho;Kim, Seung;Koh, Hong;Lee, Yoo Min;Lee, Jee Hyun;Lee, Yoon;Lee, Ji-Hyuk;Lee, Hae Jeong;Jang, Hyo-Jeong;Choi, Youjin;Choi, So Yoon;Kim, Ju Young;Choe, Byung-Ho
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.23 no.1
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    • pp.49-62
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    • 2020
  • Purpose: We aimed to investigate the disease phenotype of Korean pediatric Crohn's disease (CD) patients at diagnosis according to the Paris classification by comparison with patients from the European multicenter 5-years recruitment of children with newly developed IBD (EUROKIDS registry). Methods: Korean children and adolescents who had been newly diagnosed with CD at the age of <18 years during 2013-2016 were included in this multicenter retrospective study. Disease phenotype at diagnosis was classified according to the Paris classification, and compared with the published data from the EUROKIDS study. Results: A total of 255 patients were included. The median diagnosis age was 14.7 years (range, 0.8-17.9 years). No significant difference was observed in male-to-female ratio with EUROKIDS (1.9:1 vs. 1.45:1, p=0.062). The proportion of children aged <10 years was significantly lower in Koreans (7.1% vs. 19.6%, p<0.001). Colonic disease was less prominent (10.0% vs. 27.3%, p<0.001), while upper GI involvement was more prominent in Korean children (59.3% vs. 46.2%, p<0.001). The proportion with perianal fistulizing disease at diagnosis was significantly higher in Korean patients (44.8% vs. 8.2%, p<0.001). A separate analysis of Korean patients revealed that perianal fistulizing disease at diagnosis was positively associated with male sex and body mass index z-score (odds ratio [OR]=2.12, 95% confidence interval [CI]=1.20-3.76, p=0.010; and OR=1.29, 95% CI=1.05-1.58, p=0.015, respectively). Conclusion: Approximately half of pediatric CD patients in Korea present with perianal fistulas and/or abscesses at diagnosis, which is a distinct feature of CD in Korean children and adolescents compared to their European counterparts. An underlying genetic difference between ethnicities may play a role in this expression of different phenotypes in pediatric CD.

An analysis of one-year experience of pediatric observation unit: The first report in Korea (소아 관찰병실 운영에 대한 분석)

  • Lee, Jee Young;Choi, Ui-Yoon;Lee, Soo Young;Lee, Ji-Young;Lee, Byong Chan;Hwang, Hui Sung;Mok, Hye Rin;Jeong, Dae Chul;Chung, Seung Yun;Kang, Jin Han
    • Clinical and Experimental Pediatrics
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    • v.50 no.7
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    • pp.622-628
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    • 2007
  • Purpose : While pediatric observation units (POU) have become a common practice in hospitals throughout developed countries, there has been no report about POUs in Korea so far. The aims of this study were to analyze our one-year's experience of the POU and to decide which disease entities are suitable for the POU. Methods : All children admitted from March 2006 to February 2007 to the POU at the Department of Pediatrics in Our Lady of Mercy Hospital were included in this study. Data were collected from retrospective reviews of their medical records. Results : There were a total of 1,076 POU admissions. Median age of patients was 2.4 years and median length of hospital stay 14.0 hours. The most common diagnoses were gastroenteritis (42.7%), pharyngotonsillitis (19.1%), bronchiolitis (7.8%), pneumonia (5.5%) and febrile seizure (5.2%). Overall, 7.5% of the POU patients required subsequent inpatient admissions due to hospital stays of longer than 48 hours. The disease entities that were most likely to require inpatient admission were pneumonia (17.0%), febrile seizure (12.5%) and asthma (11.5%). Diseases that allowed successful discharge from the POU were gastroenteritis (4.6%), upper respiratory tract infection (5.8%), such as otitis media and pharygnotonsillitis and seizure disorder (6.4%). Compared with the previous year when the POU was not in operation, there was a statistically significant reduction in the average length of hospital stays (from 4.69 to 3.75 days), as well as a rise in the bed turnover rate (from 78.8 to 98.2 patients/bed). Conclusion : Our study shows that the POU is efficient for the management of children with certain acute illnesses. Based on this study, we suggest that the POU be used as a new modality which links between the outpatient, inpatient, and emergency departments in the field of pediatrics in Korea.

Analysis of gonadotropin-releasing hormone (GnRH) test results in girls with precocious puberty (성조숙증 여아에서 생식샘자극호르몬분비호르몬 검사 결과의 분석)

  • Choi, Jung Yun;Kang, Hyun-Ju;Cho, Won Kyoung;Cho, Kyoung Soon;Park, So Hyun;Hahn, Seung Hoon;Jung, Min Ho;Seo, Byung Kyu;Lee, Byung Churl
    • Clinical and Experimental Pediatrics
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    • v.52 no.12
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    • pp.1377-1382
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    • 2009
  • Purpose:The gonadotropin-releasing hormone (GnRH) test results of girls with precocious puberty were analyzed to determine whether this test can efficiently and clearly differentiate between central precocious puberty (CPP) and other disorders. Methods:Clinical and laboratory data of 54 girls with precocious pubertal signs were reviewed. Intravenous GnRH test was performed with blood samples obtained at 0, 30, 60, and 90 minutes. A peak luteinizing hormone (LH) level of ${\geq}5.0IU/L$ was indicative of CPP. Results:Of the 40 girls with CPP, 36 (90.0%), 3 (7.5%), and 1 (2.5%) showed peak LH levels at 30, 60, and 90 minutes, respectively. A percentage of girls whose peak LH ${\geq}5.0IU/L$ up to 30, 60, and 90 minutes was 92.5%, 100%, and 100%, respectively. The peak LH/follicle stimulating hormone (FSH) ratio of girls with CPP was 0.89${\pm}$0.49 and was <1 in 16 of the 40 girls (40.0%). Girls with peak LH/FSH ratio of >1.0 showed higher chronological age (CA) ($8.3{\pm}0.6$ vs. $7.7{\pm}1.0$ years, P=0.033), bone age (BA) ($10.9{\pm}0.8$ vs. $9.7{\pm}1.1$ years, P=0.001), and BA-CA difference ($2.6{\pm}0.7$ vs. $2.0{\pm}0.7$ years, P=0.009) than those of girls with peak LH/FSH ratio of ${\leq}1.0$. Higher percentage of girls with peak LH/FSH ratio of >1.0 showed advanced breast development (${\geq}TannerIII$) (93.7% vs. 41.7%, P=0.001). Conclusion:LH levels after 30 and 60 minutes of intravenous GnRH administration are the most useful for diagnosing CPP in girls.

Effects of Attenuation and Scatter Corrections in Cat Brain PET Images Using microPET R4 Scanner (MicroPET R4 스캐너에서 획득한 고양이 뇌 PET 영상의 감쇠 및 산란보정 효과)

  • Kim, Jin-Su;Lee, Jae-Sung;Lee, Jong-Jin;Lee, Byeong-Il;Park, Min-Hyun;Lee, Hyo-Jeong;Oh, Seung-Ha;Kim, Kyeong-Min;Cheon, Gi-Jeong;Lim, Sang-Moo;Chung, June-Key;Lee, Myung-Chul;Lee, Dong-Soo
    • Nuclear Medicine and Molecular Imaging
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    • v.40 no.1
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    • pp.40-47
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    • 2006
  • Purpose: The aim of this study was to examine the effects of attenuation correction (AC) and scatter correction (SC) on the quantification of PET count rates. Materials and Methods: To assess the effects of AC and SC $^{18}F$-FDG PET images of phantom and cat brain were acquired using microPET R4 scanner. Thirty-minute transmission images using $^{68}Ge$ source and emission images after injection of FDG were acquired. PET images were reconstructed using 2D OSEM. AC and SC were applied. Regional count rates were measured using ROIs drawn on cerebral cortex including frontal, parietal, and latral temporal lobes and deep gray matter including head of caudate nucleus, putamen and thalamus for pre- and post-AC and SC images. The count rates were then normalized with the injected dose per body weight. To assess the effects of AC, count ratio of "deep gray matter/cerebral cortex" was calculated. To assess the effects of SC, ROIs were also drawn on the gray matter (GM) and white matter (WM), and contrast between them ((GM-WM)/GM was measured. Results: After the AC, count ratio of "deep gray matter/cerebral cortex" was increased by $17{\pm}7%$. After the SC, contrast was also increased by $12{\pm}3%$. Conclusion: Relative count of deep gray matter and contrast between gray and white matters were increased after AC and SC, suggesting that the AC would be critical for the quantitative analysis of cat brain PET data.

A Survey of Cancer Perception in the Students of Korea University (고려대학교 학생에서의 '암에 관한 인식도' 설문 조사 연구)

  • Choi, In-Ken;Seo, Hee-Yun;Sul, Hye-Ryoung;Choi, Jong-Gwon;Sung, Hwa-Jung;Park, Kyong-Hwa;Yoon, So-Young;Oh, Sang-Cheul;Seo, Jae-Hong;Choi, Chul-Won;Shin, Sang-Won;Kim, Yeul-Hong;Kim, Byung-Soo
    • Journal of Hospice and Palliative Care
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    • v.6 no.2
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    • pp.172-176
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    • 2003
  • Purpose : This study was designed to evaluate the perception of cancer in the students of Korea University. Methods : 1,000 students of Korea University were surveyed by questionnaire from March 2001 to September 2002. Nine hundred ninety two complete questionnaires were returned. The data were analyzed by descriptive statistics, and Chi-square analysis using the SPSS program. Results : 701 students (76.1%) replied that cancer is a curable disease. However, the degree of satisfaction for cancer treatment with modem clinical medicine was low (37.5%). Moreover, they thought that there was no difference between the efficacy of treatment by chemotherapy, alternative medicine, or dietary modification. But, the positive evaluation for the effectiveness of chemotherapy in the students who had cancer patients in their families was higher than that in the students without the experience of cancer patients in their families. Six hundred ninety four students (75.3%) replied positively for euthanasia in terminal cancer patients. And they thought that the dignity and the right of the patients were the most important point that should be considered in making the decision of euthanasia. Conclusions : The credibility to modem clinical medicine for cancer treatment was relatively low in the students of Korea University who had no cancer patients in their families. So, further studies and trials will be warranted to evaluate the causes of these results and improve the credibility of modem clinical medicine for cancer treatment in the general population.

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Interleukin-4 and -8 Gene Polymorphisms and Risk of Gastric Cancer in a Population in Southwestern China

  • Pan, Xiong-Fei;Wen, Ying;Loh, Marie;Wen, Yuan-Yuan;Yang, Shu-Juan;Zhao, Zhi-Mei;Tian, Zhi;Huang, He;Lan, Hui;Chen, Feng;Soong, Richie;Yang, Chun-Xia
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.7
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    • pp.2951-2957
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    • 2014
  • Background: Gastric carcinogenesis is a complicated process that involves environmental and genetic factors like interleukin-4 (IL-4) and IL-8. Single nucleotide polymorphisms in their genes are associated with changed levels of gene expression. Here, we investigated the association between IL4-590 C>T and IL8-251T>A and gastric cancer (GC) risk in Sichuan of Southwestern China. Materials and Methods: We surveyed the research subjects using a self-designed questionnaire with questions on demographic factors and putative risk factors. Approximately 2-5ml of whole blood was collected after field survey to analyze IL4-590 C>T and IL8-251T>A genotypes using MALDI-TOF MS. Results: Our study recruited 308 pairs of GC patients and controls, including 224 (72.7%) men and 84 (27.3%) women in each group. There were 99 cardia and 176 noncardia GC patients in the case group. The case and control groups had an average age of $57.7{\pm}10.6$ ($mean{\pm}SD$) and $57.6{\pm}11.1$ years. GC patients reported a significantly greater proportion of family history of cancer (29.9% vs 10.7%, p<0.01) and drinking (54.6% vs 43.2%, p<0.01) than did controls. Variant genotypes of IL-4-590 C>T and IL-8-251 T>A were not associated with overall GC risk (adjusted OR, 0.89; 95%CI, 0.61-1.28 for CT or CC vs TT; adjusted OR, 1.14; 95%CI, 0.86-1.79 for TA or AA vs TT). Stratification analysis of two SNPs for risk by subsites only found that variant IL-8-251 TA or AA genotype was associated with increased noncardia GC risk (adjusted OR, 2.58; 95%CI, 1.19-5.57). We did not observe interactions between the IL-8-251 T>A genotype and smoking (adjusted OR, 0.38; 95%CI, 0.08-1.79) or drinking (adjusted OR, 0.36; 95%CI, 0.08-1.65) for risk of noncardia GC. Conclusions: Our data indicate no association between the two SNPs of IL-4-590 and IL-8-251 with overall GC risk, while the IL-8-251 TA or AA genotype conferred risk of cardia GC. Our findings contribute to the evidence body for risk of SNPs associated with the development of gastric cancer in this region.

Effect of Toothbrushing Facilities on PHP index and Oral Health-related Behaviors in Elementary schools (양치시설 여부에 따른 초등학생의 구강환경관리능력 및 구강건강관리 행태 변화)

  • Hwang, Yoon-Suk;Kim, Kwang-Soo;Jung, Jae-Yeon;Yoo, Young-Jae;Kim, Soo-Hwa;Lim, Mi-Hee
    • The Journal of Korean Society for School & Community Health Education
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    • v.14 no.3
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    • pp.27-40
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    • 2013
  • Objectives: This study was conducted at a request for cooperation through an analysis of the effect of toothbrushing facilities at a public health center in Seongdong-gu. Also, with the aim of furnishing basic data to the proposal of a program for improving the ability of school aged children in managing oral cavities and developing a correct toothbrushing habit, the study conducted an investigation of how toothbrushing facilities affect change in the oral environment management ability and behavior of oral health care. Methods: From among elementary schools located in Seongdong-gu, Seoul the study selected A Elementary School where toothbrushing facilities were established and have been operated since 2008, B Elementary School in an adjacent region within the jurisdiction of Seongdong-gu where the demographic environment was similar among schools where toothbrushing facilities were newly established in 2012, and C Elementary School without toothbrushing facilities. Then the study was aimed at first grade students of the schools and an investigation was carried out from April to December 2012. Through a dental checkup, the study evaluated the teeth and periodontal health conditions, and a test of the oral environment management ability was undertaken three times. Regarding change of oral health care behavior, the study carried out a self-recording survey. Results: 1. Concerning decayed and filled tooth(dft) and decayed and filled surface (dfs), A Elementary School where toothbrushing facilities have been established and operated from the past showed a relatively lower decayed, missing, and filled teeth index than B Elementary School where toothbrushing facilities were established in 2012 or C Elementary School without toothbrushing facilities; however, there was no significant difference (p>0.05). For CPI, there was no significant difference by school; however, in looking into the difference between boy students and girl students, Code (0) was discovered higher in boy students whereas Code (1) was shown higher in girl students. 2. In the PHP index test in accordance with the existence of toothbrushing facilities before the installation of toothbrushing facilities, for A Elementary School where toothbrushing facilities have been established and operated from the past, the school recorded 4.28 points whereas B Elementary School where the facilities were established in 2012 recorded 3.51 points. Meanwhile C Elementary School without the facilities posted 4.30 points. Therefore there was a statistically significant difference according to the existence of toothbrushing facilities (p<0.05). 3. In a comparison of teeth health care behavior according to the existence of toothbrushing facilities, the number of answers that the respondent did not brush their teeth after lunch over the past one week was higher in B Elementary School and C Elementary School where there were no toothbrushing facilities. Regarding the average number of brushing after lunch for one week, it was discovered higher in A Elementary School (p<0.01). 4. In change of teeth health care behavior before and after the establishment of toothbrushing facilities, the case of answering that the respondents did not brush their teeth after lunch for one week increased more after establishment than before establishment. Also the average number of teeth brushings after lunch for one week decreased further after the establishment of toothbrushing facilities; however, it did not show a significant difference (p>0.05). One of the reasons that they do not brush their teeth, "the lack of a place", decreased significantly after establishment than before establishment (p<0.05), whereas the answer, "because their friends do not brush their tooth" increased greatly after establishment than before establishment; however, there was no significant difference (p>0.05). 5. In the comparison of the degree of knowledge about dental health according to the existence of toothbrushing facilities, the degree of knowledge about dental health was shown significantly higher in A Elementary School with toothbrushing facilities than in B Elementary School and C Elementary School where there were no toothbrushing facilities (p<0.01). Conclusions: Given the above results, it is difficult to attract change in behavior only with an environmental improvement; therefore, it is deemed necessary to develop an educational program that will help children to make a habit of oral health care not only through a school but also through a related policy and financial support of government organizations as well as the construction of the basis of a systematic and consistent cooperative system with relevant organizations.

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The Difference of Lung Cancer Prevalence According to Smoking Habits (흡연 습관에 따른 폐암 발생의 차이)

  • Park, Moo Suk;Chung, Jae Ho;Jung, Jae Hee;Kim, Young Sam;Kim, Se Kyu;Jee, Sun Ha;Chang, Joon;Kim, Sung Kyu
    • Tuberculosis and Respiratory Diseases
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    • v.55 no.3
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    • pp.267-279
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    • 2003
  • Background : The purpose of this study was to analyze the smoking habits in patients with lung cancer and identify any difference of prevalence according to histologic types of lung cancer. Methods : The data were calculated by total amounts of tar and nicotine inhaled during the whole lifetime according to variation of smoking habit. This study was to investigated any difference of prevalence in lung cancer according to smoking habits. The subjects comprised 150 lung cancer cases and 300 hospital control cases that were matched by age and sex. Smoking habits during the whole lifetime were surveyed by standardized questionnaire. Odds ratios were estimated by unconditional logistic regression analysis. Results : There were 104 male and 34 female lung cancer cases. By histologic type, there were 53 cases of squamous cell carcinoma, 67 of adenocarcinoma and 30 of small cell lung carcinoma. The differences between lung cancer cases and controls according to smoking habits were total duration of smoking, total pack years of smoking and number of cigarettes smoked per day during the previous two years. The odds ratio were higher in Kreyberg I, but not in Kreyberg II, for the longer duration of smoking, the greater total pack years of cigarettes consumed, the more cigarettes smoked per day during the previous two years, the longer duration on non-filter smoking, the earlier life cases who began to smoke, and the higher amounts of calculated total tar and nicotine inhaled over the whole lifetime. When we added grade of inhalation to calculation of amounts of tar and nicotine inhaled over the lifetime, the odds ratios of total inhalation amounts of tar and nicotine were as high as those the without them. Conclusions : This study reconfirmed that smoking habits were strongly associated with lung cancer and that there were different associations between smoking habits and histologic types of lung cancer. In particular, calculations of total tar and nicotine amounts inhaled over the whole lifetime were calculated for the first time in trials from lung cancer epidemiologic studies.

Analysis of The Adverse Events Following Immunization of the National Compensation Program and the Surveillance System in Korea, 1995~2000 (1995~2000년 예방접종 후 이상반응 사례 분석 - 국가보상 신청 사례와 예방접종 후 이상반응 감시체계 보고 사례를 중심으로 -)

  • Lee, Hong Jue;Sohn, Young Mo;Kim, Joung Soon;Kim, Young Taek;Lee, Jong Gu;Choi, Bo Youl
    • Pediatric Infection and Vaccine
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    • v.8 no.2
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    • pp.135-149
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    • 2001
  • Purpose : To describe the characteristics of the claims for compensation and the methodology of investigations concerning adverse events following immunization(AEFI) and epidemiologic characteristics of the AEFI. From these results, to give a suggestion for improving AEFI surveillance system, consequently to stabilize National Immunization Program. Methods : Totally 61 cases were reported to the National Compensation Program and surveillance system of AEFI from 1995 to 2000. Documents from National Compensation Program, medical records and epidemiologic investigation reports of the cases were collected and analyzed. Results : The number of reported cases was 12 in 1995, 3 in 1996, 12 in 1998, 5 in 1999, and 29 in 2000, respectively. That of deaths was 24(39.3%) and 16 cases among them were autopsied(66.7%). That of claims for compensation was 36 and 17 cases among them were compensated(47.2%). The proportion of claim was lower in 1999~2000 than in 1995~1998, but proportion of compensation increased. Although proportion of investigation was lower in 1999~2000 than in 1995~1998, cases which were not epidemiologically investigated, decreased. The epidemiologic investigation launched within 24 hours after report increased from 25% in 1995~1998 to 48.3% in 1999~2000. Among reported cases, the number of boys was 31(50.8%), and 75.4% of subjects were from 2 months to 24 months old. 78.4% of adverse events occurred within one week after immunization. Conclusion : We can find that surveillance system of AEFI and quality of epidemiologic investigation has been somewhat improved. But, it seems that severe cases were more frequently reported than moderate or mild cases, and reporting rate was lower than that of other developed countries. We could not identify the detailed epidemiologic characteristics of AEFI due to these limitation of data. To achieve the stabilization of National Immunization Program, reporting rate and the quality of investigation should be improved.

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