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Diagnostic Performance of HPV E6/E7 mRNA and HPV DNA Assays for the Detection and Screening of Oncogenic Human Papillomavirus Infection among Woman with Cervical Lesions in China

  • Wang, Hye-young;Lee, Dongsup;Park, Sunyoung;Kim, Geehyuk;Kim, Sunghyun;Han, Lin;Yubo, Ren;Li, Yingxue;Park, Kwang Hwa;Lee, Hyeyoung
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.17
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    • pp.7633-7640
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    • 2015
  • Background: Human papillomavirus (HPV) is the most common sexually transmitted infection worldwide and it is responsible for most cases of cervical uterine cancer. Although HPV infections of the cervix do not always progress to cancer, 90% of cervical cancer cases have been found to be associated with high risk HPV (HR-HPV) infection. HPV DNA testing is widely used, along with Papanicolaou (Pap) testing, to screen for cervical abnormalities. However, there are no data on the prevalence of genotype-specific HPV infections assessed by measuring HPV E6/E7 mRNA in women representative of the Chinese population across a broad age range. Materials and Methods: In the present study, we compared the results with the CervicGen HPV RT-qDx assay, which detects 16 HR-HPV genotypes (Alpha-9: HPV 16, 31, 33, 35, 52, and 58; Alpha-7: HPV 18, 39, 45, 51, 59, and 68; and Alpha-5, 6: HPV 53, 56, 66, and 69), and the REBA HPV-ID assay, which detects 32 HPV genotypes based on the reverse blot hybridization assay (REBA) for the detection of oncogenic HPV infection according to cytological diagnosis. We also investigated the prevalence and genotype distribution of HPV infection with a total of 324 liquid-based cytology samples collected in western Shandong province, East China. Results: The overall HPV prevalences determined by HPV DNA and HPV E6/E7 mRNA assays in this study were 79.9% (259/324) and 55.6% (180/324), respectively. Although the positivity of HPV E6/E7 mRNA expression was significantly lower than HPV DNA positivity, the HPV E6/E7 mRNA assay showed greater specificity than the HPV DNA assay (88.6% vs. 48.1%) in normal cytology samples. The prevalence of Alpha-9 (HPV 16, 31, 33, 35, 52, and 58) HPV infection among these women accounted for up to 80.3% and 76.1% of the high-grade lesions detected in the HPV mRNA and DNA tests, respectively. The HR-HPV genotype distribution, based on HPV DNA and E6/E7 mRNA expression by age group in patients with cytologically confirmed lesions, was highest in women aged 40 to 49 years (35.9% for cytologically confirmed cases, Pearson correlation r value=0.993, p<0.001) for high-grade lesions. Among the oncogenic HR-HPV genotypes for all age groups, there was little difference in the distribution of HPV genotypes between the HPV DNA (HPV -16, 53, 18, 58, and 33) and HPV E6/E7 mRNA (HPV -16, 53, 33, 58, and 18) assays. HPV 16 was the most common HPV genotype among women with high-grade lesions. Conclusions: Our results suggest that the HPV E6/E7 mRNA assay can be a sensitive and specific tool for the screening and investigation of cervical cancer. Furthermore, it may provide useful information regarding the necessity for early cervical cancer screenings and the development of additional effective HPV vaccines, such as one for HPV 53 and 58. Additionally, gaining knowledge of HPV distribution may also inform us about ecological changes in HPV after the vaccination.

Effect of Nitrogen Fertilizer Rates on Rice Quality in Mid-mountainous Area (질소시비량이 중산간지 고품질 쌀 생산에 미치는 영향)

  • Kang J.R.;Kim J.T.;Beg I.Y.;Kim J.I.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.50 no.spc1
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    • pp.37-40
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    • 2005
  • In order to find out better cultivation practices in mid-mountainous area to produce high quality rice, an experiment involving seven nitrogen fertilizer rates and three varieties was carried out, during past two years$(2003\~2004)$. There was an interaction effect on palatability of milled rice between nitrogen fertilizer rate and variety. The palatability of Junambyeo grown by the treatment of nitrogen fertilizer yale wasn't different across the rates significantly, but Hwayongbyeo's palatability grown by 0 kg/ha nitrogen treatment was better than 50, 70,170 kg/ha, and Sangmibyeo's palatability grown by 70 kg/ha nitrogen treatment was better than 90, 110, 140, 170 kg/ha. Amylose content of milled rice was showed without a significant difference between nitrogen fertilizer rate, but in between variety, the amylose content of Sangmibyeo was lower than Hwayongbyeo's, also Hwayongbyeo's amylose content was lower than Junambyeo's. Protein content of milled rice was showed an interaction effect between nitrogen fertilizer rate and variety. The protein content of Hwayongbyeo and Junambyeo grown by the treatment of nitrogen fertilizer rate wasn't different across the fertilizer yale significantly, but Sangmibyeo's protein content grown by 0 kg/ha nitrogen treatment was lower than the rest of treatments, and it's protein content grown by 50, 70, 90 and 110 kg/ha nitrogen treatment was lower than 140 and 170 kg/ha. Bead rice yield of milled rice grown by lower nitrogen fertilizer rate, was lower than that grown by higher nitrogen fertilizer rate. In a conclusion gathering all above the results to produce high quality rice in Mid-mountainous area, the applicative nitrogen fertilizer rate was 70 kg per ha for Sangmibyeo, 110 kg/ha for Hwayongbyeo and 140 kg/ha for Junambyeo, in considering with head rice yield, palatability, amylose${\cdot}$protein content and productivity.

Development of a Method for Analyzing the Nicotine Content in Synthetic Flavoring Substances as Unauthorized E-cigarette Liquid by Using HPLC (전자담배 액상 충진제와 유사한 합성착향료 중 니코틴의 HPLC 분석법 개발)

  • Kim, Jae-Young;Lee, Sang-Mok;Chang, Moon-Ik;Cho, Yoon-Jae;Lee, Han-Jin;Chae, Young-Sik;Rhee, Gyu-Sik
    • Korean Journal of Food Science and Technology
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    • v.45 no.6
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    • pp.693-699
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    • 2013
  • A simple, sensitive, and specific method for quantifying the nicotine content of synthetic favoring substances (SFS) was developed using high performance liquid chromatography (HPLC) with a photo-diode array detector (PDA). Nicotine was extracted from SFS samples by using an acid-base liquid-liquid extraction method with dichloromethane and distilled water. The nicotine content was quantified by HPLC/PDA (261.9 nm) with a $C_{18}$ column under a gradient of 10% acetonitrile with 20 mM ammonium formate (ammonia solution adjusted to pH 8.7) to 100% acetonitrile. The calibration curve, analyzed from concentration standards between 0.1 to 2 mg/L, presented linearity with a correlation coefficient ($r^2$)>0.9999. The limit of quantitation (LOQ) of nicotine in SFS was 0.4 mg/kg, and the average recoveries ranged from 76.4% to 96.3%. The repeatability of measurements, expressed as the coefficient of variation (CV%), ranged from 1.74 to 5.12%. This newly developed method for nicotine quantification in SFS can be considered an analytical method with an acceptable level of sensitivity and repeatability.

Effect of Acupuncture on the Expressions of Neuropeptide Y and Leptin Receptor in the Hypothalamus of Food-deprived Rats (침치료가 굶긴 쥐 시상하부에서 neuropeptide Y(NPY)와 leptin receptor(LR)의 발현에 미치는 영향에 대한 실험적 연구)

  • Kim, Mi-A;Jung, Woo-Sang;Moon, Sang-Kwan;Kim, Young-Suk;Kim, Chang-Ju;Cho, Ki-Ho
    • The Journal of Internal Korean Medicine
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    • v.34 no.2
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    • pp.113-121
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    • 2013
  • Objectives : This study aimed to find out whether acupuncture at various acupoints shows any effects on appetite by regulating neurotransmitters in the hypothalamus through the expression of NPY and LR in the PVN via immunohistochemistry. Methods : Male Sprague-Dawley rats were divided into eight groups of five mice each. The rats in the acupuncture groups were treated with acupuncture at respective acupoints, twice per a day for 3 days. The animals were sacrificed 72 h after commencement of the experiment, the brains being dissected into serial coronal sections. Expressions of neuropeptide Y and leptin receptor in the hypothalamus were assessed by immunohistochemistry. Results : NPY expression in the PVN was enhanced and LR expression in the PVN was decreased by food deprivation. NPY immunoreactivity in the PVN of the food-derived rats was decreased by acupuncture at the auricular acupoint, Zusanli-acupoint, a and non-acupoint. However, acupuncture at the auricular acupoint showed most potent suppressing effect on NPY expression in the PVN of food-deprived rats. LR expression in the PVN decreased following food deprivation, and auricular acupuncture increased LR expression in the PVN of food-deprived rats. In normal conditions (fed state), LR expression in the PVN was not changed by acupuncture treatment at several sites. Conclusions : From this study, we have shown that acupuncture at the auricular acupoint exerts the most potent appetite suppressing effect on the food restriction state.

Dynamic Network Loading Model based on Moving Cell Theory (Moving Cell Theory를 이용한 동적 교통망 부하 모형의 개발)

  • 김현명
    • Journal of Korean Society of Transportation
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    • v.20 no.5
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    • pp.113-130
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    • 2002
  • In this paper, we developed DNL(Dynamic Network Loading) model based on Moving cell theory to analyze the dynamic characteristics of traffic flow in congested network. In this paper vehicles entered into link at same interval would construct one cell, and the cells moved according to Cell following rule. In the past researches relating to DNL model a continuous single link is separated into two sections such as running section and queuing section to describe physical queue so that various dynamic states generated in real link are only simplified by running and queuing state. However, the approach has some difficulties in simulating various dynamic flow characteristics. To overcome these problems, we present Moving cell theory which is developed by combining Car following theory and Lagrangian method mainly using for the analysis of air pollutants dispersion. In Moving cell theory platoons are represented by cells and each cell is processed by Cell following theory. This type of simulation model is firstly presented by Cremer et al(1999). However they did not develop merging and diverging model because their model was applied to basic freeway section. Moreover they set the number of vehicles which can be included in one cell in one interval so this formulation cant apply to signalized intersection in urban network. To solve these difficulties we develop new approach using Moving cell theory and simulate traffic flow dynamics continuously by movement and state transition of the cells. The developed model are played on simple network including merging and diverging section and it shows improved abilities to describe flow dynamics comparing past DNL models.

Determination of Short Term Prognosis Among Chronic Obstructive Lung Disease with Acute Respiratory Failure According to Simplified Acute Physiology Score (만성 폐쇄성 폐질환에서 급성 호흡부전 발생시 Simplifed Acute Physiology Score에 따른 단기예후의 평가)

  • Lee, Sang-Pyo;Sung, Yun-Up;Kim, Sang-Hoon;Kim, Bong-Sik;Kim, Young-Jun;Park, In-Won;Choi, Byung-Whui;Hue, Sung-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.40 no.5
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    • pp.532-539
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    • 1993
  • Background: Physician's estimates of patient survival often influence clinical decisions, especially those near the end of life. In addition. clinical decisions frequently reflect trade-offs between morbidity and length of survival. As a result, accurate estimates of survival can be extremely useful in clinical decision. When the episode of acute respiratory failure in chronic obstructive lung disease, evaluation of the severity of the condition and short term prognosis is difficulit based on the available clinical or paraclinical data at the time of admission. Method: In this study, we performed a retrospective study in Chung Ang University Hospital, 74 patients (51 males, 23 females), who were hospitalized with chronic obstructive lung disease with acute respiratory failure from 1980 to 1992. We evaluated these patients to determine lung prognostic factors at time of admission in the Intensive Care Unit (ICU) that predict short term survival, and to determine the possible application of the Simplified Acute Phsiology Score (SAPS) to this population, All patients were treated with similar regimen during the hospitalization. Results: The results were as follows: 1) Hospital mortality was 34%(25/74 patients) and surival rate was 66%(49/74 patients) in COPD with acute respiratory failure. The prognosis of the older age was much poorer than those of the young age. 2) There was no difference in mortality according to the results of basal pulmonary test and arterial blood gas analysis. 3) The SAPS at admission was higher in those patients who expired(10.8) than the survived(6.5), and there was positive correlation between SAPS and mortality (r=0.91, p<0.05). 4) Prognostic factors in acute respiratory failure complicating COPD which were identifiable at time of admission to the ICU were as follows: cachexia, encephalopathy, serum creatinine and phosphate. Conclusion: In conclusion, the SAPS might have a good prognostic value for determination of short term survival among chronic obstructive lung disease with acute respiratory failure.

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Study on Exfoliation and Recovery of Anesthetized Yound Abalones, Haliotis discus hannai Treated with Ethy-p-aminobenzoate and Freshwater in Different Temperatures of Sea water (온도구간별 파라아미노안식향산에틸과 담수에 의한 양식산 참전복 (Haliotis discus hannai) 치패의 박리 및 마취회복에 관한 연구)

  • 최상덕;정성채;김호진;공용근;백재민;최규정
    • Journal of Aquaculture
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    • v.10 no.3
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    • pp.281-288
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    • 1997
  • We investigated the optimal concentratin of ethyl-p-aminobenzoate for the exfoliation and recovery of young abalone, Haliotis discus hannai in according to different water temperatures, for the purpose of preventing the damage of shell and muscle to ecfoliated from shelter. In the 14$^{\circ}C$ water temperature, young abalones were exfoliated after 16, 35, 35 and 35 minutes in 150, 100, 75 and 50ppm concentration of ethyl-p-aminobenzoate, and were recovered after 100, 60, 30 and 30 minutes, respectively. Exfoliation rate of abalone were 100% except for 50 ppm (80%) and recovery rate were 100% of all concentration. In the $18^{\circ}C$ water temperature, young abalones were exfoliated after 4, 4, 6, 8, 8 and 12 munutes in 300, 200, 150, 100, 75 and 50ppm concentration of ethyl-p-aminobenzoate, and were recovered after 210, 180, 90, 60, 30, 20 and 20 minutes, respectively. Exfoliation rate of abalone were 100%, and recovery rate were 100% except for 200 and 300ppm (90%). In the $24^{\circ}C$ water temperature, young abalones were exfoliated after 8, 10, 10 and 12 minutes in 150, 100, 75 and 50ppm concentration of ethyl-p-aminobenzoate, and were recovered after 70, 50, 30 and 20 minutes, respectively. Exfoliation and recovery rate of abalone were 100%. In the 18$^{\circ}C$water temperature, exfoliation rate that treated with freshwater during 20 minute were 80, 50, 30 and 5% in 100, 75, 50 and 25% of fresh water, and recovery after 60, 15, 10 and 2 minutes, respectively and recovery arate were 100% except of r 100% freshwater. In this study, we suggest the reslults that the exfoliation and recovery by ethly-p-aminobenzoate were more effected in $18^{\circ}C\;and\;24^{\circ}C$ of sea water temperature than those of $14^{\circ}C$. The optimal concentration of ethyl-p-aminobenzoate was 50ppm at those water temperature. We raised 20 individual of young abalones at water temperature of $16^{\circ}C$ in the 1$\ell$ o ftnk and checked the variatin of dissolved oxygen (DO) by respiration of abalones that treated with 75ppm of ethyl-p-aminobenzoate. Before anesthetizion, DO were 6.17~6.20mg/$\ell$ and slowly decreased. But after 60 minutes, DO decreasing were stopped in 5.42~5.46mg/$\ell$. On the other hand, the control was continuously decreased and 5.27mg/$\ell$ after 60 minutes. The heartbeats of abalones were 33~45/minute in the water temperature of $18^{\circ}C$, but that treated with 100 ppm concentration of ethyl-p-aminobenzoate during 60 minutes, was 0/minute. And heartbeats of recovered abalones from anesthetizion were 29~43/minute.

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Use of Real-Time PCR and Internal Standard Addition Method for Identifying Mixed Ratio of Chicken Meat in Sausages (Real-Time PCR과 Internal Standard Addition법을 이용한 돼지고기 소시지에 혼합된 닭고기의 정량)

  • Lee, Namrye;Joo, Jae-Young;Yeo, Yong-Heon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.9
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    • pp.1097-1105
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    • 2017
  • This study examined how much chicken meat was in sausage made with pork. Both real-time polymerase chain reaction (PCR) and internal standard addition were used. Fifty ng of chicken DNA was added to the sausages as an internal standard. The addition of standard DNA increased the amplification efficiency of PCR and confirmed the possibility of quantitative analysis. A QIAamp DNA Micro Kit was used to improve the DNA recovery and amplification efficiency. The density of template DNA and primer were suitable for $3.0{\sim}5.0{\mu}L$ and $0.5{\mu}L$, respectively. Each DNA of pig and chicken was diluted in 10-fold from steps 50 ng to 0.05 ng. The detection limit of both pig and chicken meat was more than 0.05 ng and the correlation coefficient of the standard curve was at least 0.98. The result of the quantitative analysis after heat treatment of 3 samples of pigs and chickens mixed at 70:30 showed a 5.7% difference (64.3:35.7) between the expected value and measured value. The quantitative value was changed by affecting the DNA according to the heat treatment ($70^{\circ}C$, 10 min). An analysis of the pork and chicken content in sausages showed that it was difficult to detect chicken meat and the quantitative value of DNA according to the Ct value was very low. On the other hand, when adding standard material (50 ng of chicken DNA) to the sausages, the Ct value decreased gradually with increasing chicken mixing ratio. Thus, the mixing ratio of chicken in sausages could be estimated.

Growth Performance, Humoral Immune Response and Carcass Characteristics of Broiler Chickens Fed Alkali Processed Karanj Cake Incorporated Diet Supplemented with Methionine

  • Panda, K.;Sastry, V.R.B.;Mandal, A.B.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.5
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    • pp.677-681
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    • 2005
  • A study was conducted to see the effect of dietary incorporation of alkali (1.5% NaOH, w/w) processed solvent extracted karanj cake (SKC) supplemented with methionine on growth performance, humoral immune response and carcass characteristics of broiler chickens from 0 to 8 weeks of age. One hundred and twenty, day- old broiler chicks were wing banded, vaccinated against Marek' disease and distributed in a completely randomized design (CRD) into 3 groups of 40 chicks each, which was further replicated to 4 and fed on diet containing soybean meal and those of test groups were fed diets containing alkali (1.5% NaOH) treated SKC partially replacing soybean meal nitrogen of reference diet (12.5%) without or with supplementation of methionine (0.2%). Individual body weight of chicks and replicate-wise feed intakes were recorded at weekly intervals throughout the experimental period. Feed consumption from 1 to 14, 28, 42 and 56 d of age was recorded for each replicate and feed conversion efficiency (weight gain/feed intake) for the respective period was calculated. Mortality was monitored on daily basis. On 28$^{th}$ day of experimental feeding, two birds of each replicate in each dietary group (8 birds/diet) were inoculated with 0.1 ml of a 1.0% suspension of sheep red blood cells (SRBC) and the antibody titre (log 2) was measured after 5 days by the microtitre haemmagglutination procedure. After 42 days of experimental feeding, a retention study of 4 days (43-47 d) duration was conducted on all birds to determine the retention of various nutrients such as DM, N, Ca, P and GE. On 43$^{rd}$ day of experimental feeding, one representative bird from each replicate of a dietary treatment (4/dietary group) was sacrificed, after fasting for two hours with free access to water, through cervical dislocation to observe the weight of dressed carcass, primal cuts (breast, thigh, drumstick, back, neck and wing), giblet (liver, heart and gizzard), abdominal fat and digestive organs. The body weight gain of chicks fed reference diet and those fed diet incorporated with NaOH treated SKC (12.5% replacement) with or without methionine supplementation was comparable during 0 to 4 weeks of age. However, dietary incorporation of alkali processed SKC replacing 12.5% nitrogen moiety of soybean meal resulted in growth retardation, subsequently as evidenced by significantly (p<0.05) lowered body weight gain during 0 to 6 weeks of age in birds fed diet incorporated with alkali processed SKC at 6.43% without methionine as compared to those supplemented with methionine or reference diet. Dietary incorporation of alkali (1.5% NaOH) processed SKC replacing 12.5% of soybean meal nitrogen in the diet of broiler chickens had no adverse effect on feed conversion ratio during all the weeks of experimental feeding. The humoral immune response (HIR) as measured by the antibody titre in response to SRBC inoculation was comparable among all the dietary groups. No significant difference in the intake and retention of DM, N, Ca, P or GE was noted among the chicks fed reference and alkali processed SKC incorporated diets with or without methionine supplementation. None of the carcass traits varied significantly due to dietary variations, except the percent weight of liver and giblet. The percent liver weight was significantly (p<0.05) higher in the birds fed diet incorporated with alkali processed SKC as compared to that in other two groups. Thus solvent extracted karanj cake could be incorporated after alkali (1.5% NaOH, w/w) processing at an enhanced level of 6.43%, replacing 12.5% of soybean meal nitrogen, in the broiler diets up to 4 weeks of age, beyond which the observed growth depression on this diet could be alleviated by 0.2% methionine supplementation.

Association between Tumor Necrosis Factor-$\alpha$ Gene Polymorphism and Bronchopulmonary Dysplasia in Preterm Infants (조산아 기관지폐이형성증과 Tumor Necrosis Factor-$\alpha$ 유전자 다형성과의 연관성)

  • Jo, Heui-Seung;Chang, Yoon-Hwan;Kim, Han-Suk;Kim, Byeong-Il;Choi, Jung-Hwan
    • Neonatal Medicine
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    • v.18 no.1
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    • pp.42-48
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    • 2011
  • Purpose: Several factors including prolonged inflammatory response are thought to contribute to the pathogenesis of bronchopulmonary dysplasia (BPD). The clinical findings can be explained by an increased production of proinflammatory cytokines such as tumor necrosis factor alpha (TNF-$\alpha$ ). We investigated the relationship between susceptibility to BPD and TNF-$\alpha$ promoter polymorphisms to identify genetic factors of the disease. Methods: Thirty-eight preterm infants who had developed BPD and 55 controlled infants with a birth weight <1,500 g were analyzed for TNF-$\alpha$ genotypes. The alleles of five promoter sites (-1031/-863/-857/-308/-238) of TNF-$\alpha$ gene were determined using $Taqman^{(R)}$-based allelic discrimination assays. Results: Gestational age ($27^{+5}{\pm}2^{+0}$ wk vs. $29^{+2}{\pm}1^{+4}$ wk, P<0.0001) and birth weight (990${\pm}$270 g vs. 1,220${\pm}$230 g, P<0.0001) were lower in the BPD group compared to the control group. The incidence of respiratory distress syndrome (71.1% vs. 49.1%, P=0.035) and patent ductus arteriosus (71.1% vs. 50.9%, P=0.052) was higher in the BPD group compared to the control group. The frequencies of the alleles and genotypes of five promoter sites (-1031/-863/-857/-308/-238) of TNF-$\alpha$ gene did not show differences between the BPD group and the control group. Conclusion: TNF-$\alpha$ promoter polymorphisms are not associated with susceptibility to BPD in Korean preterm infants.