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Relationship between porcine miR-20a and its putative target low-density lipoprotein receptor based on dual luciferase reporter gene assays

  • Ding, Yueyun;Zhu, Shujiao;Wu, Chaodong;Qian, Li;Li, DengTao;Wang, Li;Wan, Yuanlang;Zhang, Wei;Yang, Min;Ding, Jian;Wu, Xudong;Zhang, Xiaodong;Gao, Yafei;Yin, Zongjun
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.7
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    • pp.922-929
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    • 2019
  • Objective: Mutations in low-density lipoprotein receptor (LDLR), which encodes a critical protein for cholesterol homeostasis and lipid metabolism in mammals, are involved in cardiometabolic diseases, such as familial hypercholesterolemia in pigs. Whereas microRNAs (miRNAs) can control LDLR regulation, their involvement in circulating cholesterol and lipid levels with respect to cardiometabolic diseases in pigs is unclear. We aimed to identify and analyze LDLR as a potential target gene of SSC-miR-20a. Methods: Bioinformatic analysis predicted that porcine LDLR is a target of SSC-miR-20a. Wild-type and mutant LDLR 3'-untranslated region (UTR) fragments were generated by polymerase chain reaction (PCR) and cloned into the pGL3-Control vector to construct pGL3 Control LDLR wild-3'-UTR and pGL3 Control LDLR mutant-3'-UTR recombinant plasmids, respectively. An miR-20a expression plasmid was constructed by inserting the porcine premiR-20a-coding sequence between the HindIII and BamHI sites in pMR-mCherry, and constructs were confirmed by sequencing. HEK293T cells were co-transfected with the miR-20a expression or pMR-mCherry control plasmids and constructs harboring the corresponding 3'-UTR, and relative luciferase activity was determined. The relative expression levels of miR-20a and LDLR mRNA and their correlation in terms of expression levels in porcine liver tissue were analyzed using reverse-transcription quantitative PCR. Results: Gel electrophoresis and sequencing showed that target gene fragments were successfully cloned, and the three recombinant vectors were successfully constructed. Compared to pMR-mCherry, the miR-20a expression vector significantly inhibited wild-type LDLR3'-UTR-driven (p<0.01), but not mutant LDLR-3'-UTR-driven (p>0.05), luciferase reporter activity. Further, miR-20a and LDLR were expressed at relatively high levels in porcine liver tissues. Pearson correlation analysis revealed that porcine liver miR-20a and LDLR levels were significantly negatively correlated (r = -0.656, p<0.05). Conclusion: LDLR is a potential target of miR-20a, which might directly bind the LDLR 3'-UTR to post-transcriptionally inhibit expression. These results have implications in understanding the pathogenesis and progression of porcine cardiovascular diseases.

Effect of Non-starch Polysaccharides and Resistant Starch on Mucin Secretion and Endogenous Amino Acid Losses in Pigs

  • Morel, Patrick C.H.;Melai, J.;Eady, S.L.;Coles, G.D.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.11
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    • pp.1634-1641
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    • 2005
  • Generally, dietary fibre (DF) includes lignin, non-starch polysaccharides (NSP) and resistant starch (RS). In monogastric species, low levels of dietary fibre in the diet are associated with various diseases and high levels reduce nutrient digestibilities. In this study, the effects of different types and levels of NSP (soluble: $\beta$-glucan, insoluble cellulose) and resistant starch on mucin secretion and endogenous nitrogen and amino acid losses in pigs were investigated. A total of 25 five-week-old weaner pigs (9.5 kg${\pm}$1.5 kg), were randomly allocated to each of five experimental diets. Different levels of purified barley $\beta$-glucan (BG) extract (5 or 10% of $Glucagel^{(R)}$ $\beta$-glucan, providing 4 or 8% of BG in the diet), and resistant starch (RS) (8.3 or 16.6% of Hi-$Maize^{TM}$, providing 5 or 10% RS in the diet) were substituted for wheat starch in a purified diet in which enzymatically-hydrolysed casein was the sole source of protein. The diets were fed for 21 days. No statistically significant difference between treatments (p>0.05) was observed for growth performance and organs weights. No difference in ileal starch digestibility was observed between pigs on the cellulose or $\beta$-glucan diets. However, as the level of resistant starch in the diet increased the ileal starch digestibility decreased (p<0.05). The inclusion of resistant starch in the diet (5 or 10%) did not increase mucin production when compared with the cellulose-only diet. However, as the level of beta-glucan in the diet increased, both crude mucin in the digesta dry matter and per kg dry matter intake increased (p<0.05). Pigs fed the diet containing 8% of beta-glucan had higher endogenous loss flow than those fed the diets including 5 or 10% of resistant starch or 4% of $\beta$-glucan. In conclusion, dietary inclusion of resistant starch increased the level of starch reaching the large intestine without any effect on mucin secretion, or endogenous nitrogen or amino acid losses content in the small intestine. The addition of $\beta$-glucan to a diet containing cellulose increases both mucin secretion and endogenous amino acid and nitrogen losses in the small intestine.

The 1/f Noise Analysis of 3D SONOS Multi Layer Flash Memory Devices Fabricated on Nitride or Oxide Layer (산화막과 질화막 위에 제작된 3D SONOS 다층 구조 플래시 메모리소자의 1/f 잡음 특성 분석)

  • Lee, Sang-Youl;Oh, Jae-Sub;Yang, Seung-Dong;Jeong, Kwang-Seok;Yun, Ho-Jin;Kim, Yu-Mi;Lee, Hi-Deok;Lee, Ga-Won
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.25 no.2
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    • pp.85-90
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    • 2012
  • In this paper, we compared and analyzed 3D silicon-oxide-nitride-oxide-silicon (SONOS) multi layer flash memory devices fabricated on nitride or oxide layer, respectively. The device fabricated on nitride layer has inferior electrical properties than that fabricated on oxide layer. However, the device on nitride layer has faster program / erase speed (P/E speed) than that on the oxide layer, although having inferior electrical performance. Afterwards, to find out the reason why the device on nitride has faster P/E speed, 1/f noise analysis of both devices is investigated. From gate bias dependance, both devices follow the mobility fluctuation model which results from the lattice scattering and defects in the channel layer. In addition, the device on nitride with better memory characteristics has higher normalized drain current noise power spectral density ($S_{ID}/I^2_D$>), which means that it has more traps and defects in the channel layer. The apparent hooge's noise parameter (${\alpha}_{app}$) to represent the grain boundary trap density and the height of grain boundary potential barrier is considered. The device on nitride has higher ${\alpha}_{app}$ values, which can be explained due to more grain boundary traps. Therefore, the reason why the devices on nitride and oxide have a different P/E speed can be explained due to the trapping/de-trapping of free carriers into more grain boundary trap sites in channel layer.

Comparisons of Effects of Biman-tang according to Administration Period in Childhood Obesity (비만환아에서 복약기간에 따른 비만탕의 치료효과 비교연구)

  • Lim, Youngkwern;Min, Seorim;Kim, Heeman;Hur, Kwang-Wook;Lee, Hoon;Lee, Jinsoo;Kim, Hocheol;Park, Jae-Woo;Park, Hi-Joon
    • The Journal of Pediatrics of Korean Medicine
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    • v.27 no.4
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    • pp.57-67
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    • 2013
  • Objectives The purpose of this study is to investigate the safety and effect of Biman-tang (BMT) on two different administration periods in the treatment of childhood overweight and obesity. Methods In retrospective study, 39 overweight and obese ($85^{th}$ percentile ${\leq}$ Body Mass Index (BMI)) children were treated with BMT from January 2006 to April 2013 at Korean Medical Clinic in Suwon, Korea. The primary outcomes were the changes in BMI and Obesity Index (OI) from baseline to the treatment groups. Secondary outcomes included the changes in height, weight and safety of the medicine. Comparisons of BMI, OI, height and weight between the short-period (SP) treatment group and the long-period (LP) treatment group were done by using ANCOVA. Results The change of the mean of BMI ($-0.5{\pm}0.6kg/m^2$ vs $-1.4{\pm}0.8kg/m^2$, respectively; p=0.003) and OI ($-3.6{\pm}3.9%$ vs $-9.7{\pm}4.7%$, respectively; p<0.001) showed significant reduction both in SP (n=16) and LP treatment group (n=23). The mean height showed no significant difference in both groups. The mean weight of LP treatment group showed significant reduction compared to SP treatment group ($-0.2{\pm}1.3kg$ vs $-1.6{\pm}1.6kg$, respectively; p=0.006). Most of the children were compliant to the medication and no serious adverse events were found in two groups. Conclusions These findings emphasize that BMT is effective in the treatment of childhood obesity and it requires at least 45 days of treatment for the best result.

Dough Properties and Bread Qualities of Wheat Flour Supplemented with Rice Bran (미강 첨가에 따른 밀가루 반죽물성 및 제빵적성)

  • Chang, Kyung-Hi;Byun, Gwang-In;Park, Sang-Ha;Kang, Woo-Won
    • Food Science and Preservation
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    • v.15 no.2
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    • pp.209-213
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    • 2008
  • We examined the dough properties, baking qualities, and sensory aspects of commercial hard wheat flour (Q1) that was supplemented with up to 30% rice bran flour. The loaf volumes of bread with 20% and 30% rice bran were 83% and 65% respectively, of the control (wheat flour only). However, a 5% and 10% supplement of rice bran increased loaf volume by 5.2% and 0.7% respectively, over the control. After storage of loaves for three days, bread crumbs from loaves with 5% rice bran were significantly softer than those with 10% 20% and 30% rice bran. The hardness of dough increased with the proportion of rice bran, but springiness and cohesiveness were inversely related to hardness. Addition of rice bran increased loaf pH. Rice bran decreased lightness, but significantly increased redness and yellowness in a concentration dependent manner. Sensory investigation indicated that bread with 5% rice bran was not significantly different from the control. However, more panel members preferred the appearance and flavor of bread with 5% rice bran.

The Correlation Analysis between Heart Rate Variability and Effect of Acupuncture on Obese Women (자율신경 활성도와 비만 여성 침치료 효과의 상관성 연구)

  • Kim, Je-In;Yang, Yo-Chan;Kim, Koh-Woon;Cho, Jae-Heung;Kim, Song-Yi;Park, Hi-Joon;Song, Mi-Yeon
    • Journal of Korean Medicine Rehabilitation
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    • v.26 no.4
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    • pp.85-96
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    • 2016
  • Objectives The purpose of this study was to investigate the relationship between the effects of acupuncture treatment and heart rate variability (HRV) in pre-menopausal obese women. Methods Thirty-seven obese women who met the inclusion criteria were recruited. To estimate the effects of acupuncture, obesity indices, such as body weight (BW), waist circumference (WC), hip circumference (HC), and the waist-hip ratio (WHR), were measured before and after the treatment. The HRV test was conducted before treatment and analyzed using the frequency domain method. Results The lnLF/HF ratio (natural logarithm of low frequency power/high frequency power ratio of the HRV value) before treatments was negatively correlated with differences in WC, HC, and WHR during treatment. The correlation coefficients between the lnLF/HF ratio and the differences in WC, HC, and WHR were r=-0.459 (p<0.01), r=-0.327 (p<0.05), and r=-0.339 (p<0.05) respectively. Conclusions As the baseline ratio of sympathetic activity to parasympathetic activity decreases, WC, HC, and WHR reduction significantly increased during treatment. Further study is needed to uncover the relationship between obesity-related variables and the autonomic nervous system to predict the effect of acupuncture.

Associations of Non Alcoholic Fatty Liver with the Metabolic Syndrome and Serum Carotenoids (비알코올성 지방간과 대사증후군 및 혈청 카로테노이드와의 관련성)

  • Park, Sun-Kyun;Lee, Hyun-Jung;Lee, Duk-Hee;Lee, Sung-Kook;Chun, Byung-Yeol;Kim, Sung-Ae;Lee, Hye-Sung;Son, Hyo-Kyung;Kim, Sung-Hi
    • Journal of Preventive Medicine and Public Health
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    • v.41 no.1
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    • pp.39-44
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    • 2008
  • Objectives : This study was conducted to investigate the associations of non alcoholic fatty liver with metabolic syndrome and the serum carotenoids. Methods : This study was conducted in a general hospital in South Korea from November, 2004 to August, 2005. The study subjects were 350 sampled persons who were aged from 40 years and older (males : 180, females : 170). They were grouped into the normal, mild and severe groups according to fat accumulation in their livers, as determined by ultrasonography. We analyzed the association between non alcoholic fatty liver and metabolic syndrome by multiple logistic regression analysis and we analyzed the association between non alcoholic fatty liver and the serum carotenoids by a general linear model(ANCOVA). Results : After adjustment for the effect of potential covariates, the prevalence of metabolic syndrome was associated with fat accumulation in the liver (p trend <0.001). If the odds ratio of normal group is 1.00, then that of the mild group is 2.80 (95% C.I=1.17-6.71) and that of the severe group is 7.29 (95% C.I=2.76-19.30). The prevalence of metabolic alterations fitting the criteria of metabolic syndrome, according to the class of fat accumulation in the liver, was significantly increased, except for criteria of high blood pressure, a large waist circumference and low HDL (high density lipoprotein) cholesterol level (p trend <0.001). The level of serum ${\beta}$-carotene was decreased according to the class of fat accumulation in the liver (p trend=0.036), but the levels of serum ${\alpha}$-carotene, lycopene, ${\beta}$-cryptoxanthin and lutein were not decreased. Conclusions : This study shows that non alcoholic fatty liver was associated with metabolic syndrome and with the serum ${\beta}$-carotene level.

Effect of Hominis Placenta on cutaneous wound healing in normal and diabetic mice

  • Park, Ji-Yeun;Lee, Jiyoung;Jeong, Minsu;Min, Seorim;Kim, Song-Yi;Lee, Hyejung;Lim, Yunsook;Park, Hi-Joon
    • Nutrition Research and Practice
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    • v.8 no.4
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    • pp.404-409
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    • 2014
  • BACKGROUND/OBJECTIVES: The number of diabetic patients has recently shown a rapid increase, and delayed wound healing is a major clinical complication in diabetes. In this study, the wound healing effect of Hominis placenta (HP) treatment was investigated in normal and streptozotocin-induced diabetic mice. MATERIALS/METHODS: Four full thickness wounds were created using a 4 mm biopsy punch on the dorsum. HP was injected subcutaneously at the middle region of the upper and lower wounds. Wounds were digitally photographed and wound size was measured every other day until the 14th day. Wound closure rate was analyzed using CANVAS 7SE software. Wound tissues were collected on days 2, 6, and 14 after wounding for H/E, immunohistochemistry for FGF2, and Masson's trichrome staining for collagen study. RESULTS: Significantly faster wound closure rates were observed in the HP treated group than in normal and diabetes control mice on days 6 and 8. Treatment with HP resulted in reduced localization of inflammatory cells in wounded skin at day 6 in normal mice and at day 14 in diabetic mice (P < 0.01). Expression of fibroblast growth factor (FGF) 2 showed a significant increase in the HP treated group on day 14 in both normal (P < 0.01) and diabetic mice (P < 0.05). In addition, HP treated groups showed a thicker collagen layer than no treatment groups, which was remarkable on the last day, day 14, in both normal and diabetic mice. CONCLUSIONS: Taken together, HP treatment has a beneficial effect on acceleration of cutaneous wound healing via regulation of the entire wound healing process, including inflammation, proliferation, and remodeling.

Seasonal Variations in Primary Productivity and Pigments of Downstream Water of the Han River (한강하류수역의 기초생산과 식물플랭크톤 색소량의 년변화)

  • Choe, Sang;Chung, Tai Wha;Kwak, Hi-Sang
    • 한국해양학회지
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    • v.3 no.1
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    • pp.16-25
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    • 1968
  • This study was undertaken to assess the annual cycle of primary production and plant pigments in a downstream of the Han River. Measurements were carried out at three week intervals during April 1966 and March 1967, and ancillary data include water temperature, transparency, pH, dissolved oxygen and phytoplankton cell number. The seasonal cycle in water temperature profile shows the hihgest in the end of August with 27$^{\circ}C$, lowest in the middle of February with 0.2$^{\circ}C$. The transparency with Secchi disk reading varied from a maximum 4.0m in fall and a minimum 0.5m or less in early spring and flood season of summer. The pH of the river water varied from 6.5 to 7.3, averaged 6.91 in the surface water and 6.98 in the bottom water, showed little seasonalvariability. The dissolved oxygen in the surface water ranged from 5.93-9.64ml/L, while in the bottom water it ranged from 5.54-9.72 ml/L, and the oxygen saturation never fall below 94%. None thermal, the distribution of pH and content of oxygen, stratifications occurred. An apparent seasonal cycle of primary productivity was observed with remarkably high levels in the spring and fall, the lowest level in the winter. The range of net carbon assimilations showed 3.1-112.6 mgC/㎥/day or 15-427 mgC/㎡/day in spring, 37.0-271.2 mgC/㎥/day or 115-329 mgC/㎡/day in summer, 27.2-168.0 mgC/㎥ /day or 139-415 mgC/㎡/day in fall and 0.5-10.9 mgC/㎥/day or 5-19 mg/㎡/day in winter, respectively. Amount of chlorophyll ${\alpha}$ ranged from a minimum concentration of 0.2-0.3 mg/㎥ in the middle of February and a maximum 4.1-6.7 mg/㎥ in the middle of June. A general increase trend in chlorophyll ${\alpha}$ concentration was noted with increase of the river water temperature.

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Determination of Appropriate Sampling Time for Job Stress Assessment: the Salivary Chromogranin A and Cortisol in Adult Females

  • Hong, Ran-Hi;Yang, Yun-Jung;Kim, Sang-Yon;Lee, Won-Young;Hong, Yeon-Pyo
    • Journal of Preventive Medicine and Public Health
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    • v.42 no.4
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    • pp.231-236
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    • 2009
  • Objectives : This study was conducted to determine the appropriate sampling time of the salivary stress markers, chromogranin A (CgA) and cortisol as objective indices of job stress assessment in adult females. Methods : The subjects were 20${\sim}$39-year-old women (13 office workers, 11 sales-service workers, and 11 college students) who were eligible for the study and free of acute and chronic medical conditions. Salivary CgA and cortisol levels were determined by enzyme-linked immunosorbent assay (ELISA). Saliva samples were collected (2 $m{\ell}$ each) at 7:00, 8:00, 10:30, 12:00, 17:30, and 22:30 on a typical day. Salivary CgA and cortisol levels, according to sampling time, were compared among the three groups using general linear model. The full version of the Korean Occupational Stress Scale (KOSS), which includes socioeconomic characteristics, health behavior, workrelated characteristics, and BMI, was used to access the subjects' job stress. Multiple regression analysis of the job stressors identified by the KOSS was performed on salivary CgA and cortisol levels. Results : The salivary CgA level peaked at 7:00 (time of awakening), then decreased and were maintained at a low level throughout the day, and increased slightly at 17:30. The salivary cortisol level increased steeply within the 1st hour after awakening, followed by a gradual decrease by 12:00, and was then maintained at a low level throughout the day. The salivary cortisol levels of subjects who worked ${\leq}$5 days per week and graduated from the university were significantly lower at 8:00 (p=0.006). The salivary cortisol levels of non-smokers were significantly lower at 7:00 p=0.040) and 8:00 (p=0.003) compared to smokers. There were no significant differences in salivary CgA and cortisol levels at 10:30 and 12:00 in general characteristics. The regression coefficients on salivary CgA level were significant with interpersonal conflict at 17:30 and job insecurity at 22:30. Regression coefficients on salivary cortisol level were significant with organizational system and total job stressors at 17:30. Conclusions : We suggest that the appropriate sampling times for the salivary stress markers, CgA and cortisol, are at 7:00 (time of awakening), 8:00 (1 hour after awakening), 17:30 (early evening), and 22:30 (before sleep).