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Preparation of minor ginsenosides C-Mc, C-Y, F2, and C-K from American ginseng PPD-ginsenoside using special ginsenosidase type-I from Aspergillus niger g.848

  • Liu, Chun-Ying;Zhou, Rui-Xin;Sun, Chang-Kai;Jin, Ying-Hua;Yu, Hong-Shan;Zhang, Tian-Yang;Xu, Long-Quan;Jin, Feng-Xie
    • Journal of Ginseng Research
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    • v.39 no.3
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    • pp.221-229
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    • 2015
  • Background: Minor ginsenosides, those having low content in ginseng, have higher pharmacological activities. To obtain minor ginsenosides, the biotransformation of American ginseng protopanaxadiol (PPD)-ginsenoside was studied using special ginsenosidase type-I from Aspergillus niger g.848. Methods: DEAE (diethylaminoethyl)-cellulose and polyacrylamide gel electrophoresis were used in enzyme purification, thin-layer chromatography and high performance liquid chromatography (HPLC) were used in enzyme hydrolysis and kinetics; crude enzyme was used in minor ginsenoside preparation from PPD-ginsenoside; the products were separated with silica-gel-column, and recognized by HPLC and NMR (Nuclear Magnetic Resonance). Results: The enzyme molecular weight was 75 kDa; the enzyme firstly hydrolyzed the C-20 position 20-O-${\beta}$-D-Glc of ginsenoside Rb1, then the C-3 position 3-O-${\beta}$-D-Glc with the pathway $Rb1{\rightarrow}Rd{\rightarrow}F2{\rightarrow}C-K$. However, the enzyme firstly hydrolyzed C-3 position 3-O-${\beta}$-D-Glc of ginsenoside Rb2 and Rc, finally hydrolyzed 20-O-L-Ara with the pathway $Rb2{\rightarrow}C-O{\rightarrow}C-Y{\rightarrow}C-K$, and $Rc{\rightarrow}C-Mc1{\rightarrow}C-Mc{\rightarrow}C-K$. According to enzyme kinetics, $K_m$ and $V_{max}$ of Michaelis-Menten equation, the enzyme reaction velocities on ginsenosides were Rb1 > Rb2 > Rc > Rd. However, the pure enzyme yield was only 3.1%, so crude enzyme was used for minor ginsenoside preparation. When the crude enzyme was reacted in 3% American ginseng PPD-ginsenoside (containing Rb1, Rb2, Rc, and Rd) at $45^{\circ}C$ and pH 5.0 for 18 h, the main products were minor ginsenosides C-Mc, C-Y, F2, and C-K; average molar yields were 43.7% for C-Mc from Rc, 42.4% for C-Y from Rb2, and 69.5% for F2 and C-K from Rb1 and Rd. Conclusion: Four monomer minor ginsenosides were successfully produced (at low-cost) from the PPD-ginsenosides using crude enzyme.

A Comparison Research of Eastern and Western Medication on the Insomnia (불면증에 대한 동서의학의 약물치료 비교 분석)

  • Jung, Song-Hwa;Kim, Ji-Hwon;Lee, Hyo-Gyung;Jang, Hyun-Ho;Kim, Tae-Heon;Lyu, Yeoung-Su;Kang, Hyung-Won
    • Journal of Oriental Neuropsychiatry
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    • v.20 no.3
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    • pp.65-88
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    • 2009
  • Objectives : There has not been the comparison research of medication on the Insomnia as the complementary significance of the Oriental-Western Cooperative Therapeutic Viewpoint so far. The aim of this preliminary research is to study Eastern and Western Medication Model for the Insomnia through the comparison of Oriental Medication based on "Sang Han Lon" and "Dong yui bo gam". Methods : The author researched oriental-western viewpoints and the present treatment condition of Sleep and Sleeping Pills with reference to Korean articles issued since 1975, dissertations, journals registered or candidated on KCI, alternative medicine journals on Pubmed, and books concerning Neuropsychiatry. In the field of Oriental medical study, this study was demonstrated on "The Oriental Medical textbook of neuropsychiatry", "Sang han lon", "Dong yui bo gam" and the present research trend. Results : In Oriental Medicine, Insomnia is treated according to si-jie-bu-shui-zung (思結不睡證) type, rong-xie-bu-zu-zung (榮血不足證) type, yin-her-nae-re-zung (陰虛內熱證) type, xin-dan-her-qie-zung (心膽虛怯證) type, dam-xian-yu-jie-zung (痰涎鬱結證) type, and wei-zhong-bu-he-zung (胃中不和證) type etc. In Western Medicine, Insomnia has been treated by Barbiturate over 60's and BZ over 70's. According to recent treatment on Insomnia, Zolpidem or Zaleplon, which is short-lived and has less side effects, is used much, and tri-cyclic antidepressants are prescribed on a chronic psycho-physiological insomnia. Conclusions : Generally, Insomnia is being treated by controlling whole ways. The oriental medication is less effective for sleep induction. On the contrary, that of western medication is very effective for sleep induction, but there are many possibilities of side effects. If this two mutual therapy is conducted cooperatively, more effective medical care might be expected. Therefore, it is considered that not only the oriental medical study for sleep induction, but the study of Oriental-Western Cooperative Therapy should be preceded to treat Insomnia from now on.

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The effect of various sandblasting conditions on surface changes of dental zirconia and shear bond strength between zirconia core and indirect composite resin

  • Su, Naichuan;Yue, Li;Liao, Yunmao;Liu, Wenjia;Zhang, Hai;Li, Xin;Wang, Hang;Shen, Jiefei
    • The Journal of Advanced Prosthodontics
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    • v.7 no.3
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    • pp.214-223
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    • 2015
  • PURPOSE. To measure the surface loss of dental restorative zirconia and the short-term bond strength between an indirect composite resin (ICR) and zirconia ceramic after various sandblasting processes. MATERIALS AND METHODS. Three hundred zirconia bars were randomly divided into 25 groups according to the type of sandblasting performed with pressures of 0.1, 0.2, 0.4 and 0.6 MPa, sandblasting times of 7, 14 and 21 seconds, and alumina powder sizes of 50 and $110{\mu}m$. The control group did not receive sandblasting. The volume loss and height loss on zirconia surface after sandblasting and the shear bond strength (SBS) between the sandblasted zirconia and ICR after 24-h immersion were measured for each group using multivariate analysis of variance (ANOVA) and Least Significance Difference (LSD) test (${\alpha}$=.05). After sandblasting, the failure modes of the ICR/zirconia surfaces were observed using scanning electron microscopy. RESULTS. The volume loss and height loss were increased with higher sandblasting pressure and longer sandblasting treatment, but they decreased with larger powder size. SBS was significantly increased by increasing the sandblasting time from 7 seconds to 14 seconds and from 14 seconds to 21 seconds, as well as increasing the size of alumina powder from $50{\mu}m$ to $110{\mu}m$. SBS was significantly increased from 0.1 MPa to 0.2 MPa according to the size of alumina powder. However, the SBSs were not significantly different with the sandblasting pressure of 0.2, 0.4 and 0.6 MPa. The possibilities of the combination of both adhesive failure and cohesive failure within the ICR were higher with the increases in bonding strength. CONCLUSION. Based on the findings of this study, sandblasting with alumina particles at 0.2 MPa, 21 seconds and the powder size of $110{\mu}m$ is recommended for dental applications to improve the bonding between zirconia core and ICR.

Effects of Polyurethane Coated Urea Supplement on In vitro Ruminal Fermentation, Ammonia Release Dynamics and Lactating Performance of Holstein Dairy Cows Fed a Steam-flaked Corn-based Diet

  • Xin, H.S.;Schaefer, D.M.;Liu, Q.P.;Axe, D.E.;Meng, Q.X.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.4
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    • pp.491-500
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    • 2010
  • Three experiments were conducted to investigate the effects of polyurethane coated urea on in vitro ruminal fermentation, ammonia release dynamics and lactating performance of Holstein dairy cows fed a steam-flaked corn-based diet. In Exp. 1, a dual-flow continuous culture was run to investigate the effect of polyurethane coated urea on nutrient digestibility, rumen fermentation parameters and microbial efficiency. Three treatment diets with isonitrogenous contents (13.0% CP) were prepared: i) feedgrade urea (FGU) diet; ii) polyurethane coated urea (PCU) diet; and iii) isolated soy protein (ISP) diet. Each of the diets consisted of 40% steam-flaked corn meal, 58.5% forages and 1.5% different sources of nitrogen. PCU and FGU diets had significantly lower digestibility of NDF and ADF (p<0.01) than the ISP diet. Nitrogen source had no significant effect (p = 0.62) on CP digestibility. The microbial efficiency (expressed as grams of microbial N/kg organic matter truly digested (OMTD)) in vitro of the PCU diet (13.0 g N/kg OMTD) was significantly higher than the FGU diet (11.3 g N/kg OMTD), but comparable with the ISP diet (14.7 g N/kg OMTD). Exp. 2, an in vitro ruminal fermentation experiment, was conducted to determine the ammonia release dynamics during an 8 h ruminal fermentation. Three treatment diets were based on steam-flaked corn diets commonly fed to lactating cows in China, in which FGU, PCU or soybean meal (SBM) was added to provide 10% of total dietary N. In vitro $NH_3-N$ concentrations were lower (p<0.05) for the PCU diet than the FGU diet, but similar to that for the SBM diet at all time points. In Exp. 3, a lactation trial was performed using 24 lactating Holstein cows to compare the lactating performance and blood urea nitrogen (BUN) concentrations when cows were fed PCU, FGU and SBM diets. Cows consuming the PCU diet had approximately 12.8% more (p = 0.02) dietary dry matter intake than those consuming the FGU diet. Cows fed the PCU diet had higher milk protein content (3.16% vs. 2.94%) and lower milk urea nitrogen (MUN) concentration (13.0 mg/dl vs. 14.4 mg/dl) than those fed the FGU diet. Blood urea nitrogen (BUN) concentration was significantly lower for cows fed the PCU (16.7 mg/dl) and SBM (16.4 mg/dl) diets than the FGU (18.7 mg/dl) diet. Cows fed the PCU diet had less surplus ruminal N than those fed the FGU diet and produced a comparable lactation performance to the SBM diet, suggesting that polyurethane coated urea can partially substitute soybean meal in the dairy cow diet without impairing lactation performance.

Effects of 20-day litter weight on weaned piglets' fighting behavior after group mixing and on heart rate variability in an isolation test

  • Sun, YaNan;Lian, XinMing;Bo, YuKun;Guo, YuGuang;Yan, PeiShi
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.2
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    • pp.267-274
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    • 2017
  • Objective: The objective of this study was to investigate the effect of 20-day litter weight on behavior and heart rate variability (HRV) of piglets under stress. Methods: Forty four original litters were categorized as high litter weight (HW) litters (n = 22) and low litter weight (LW) litters (n = 22) by 20-day litter weight. From each original HW litter, three males and three females were randomly selected after weaning and the 12 piglets from two original litters with similar age of days were regrouped into one new high litter weight (NHW) litter (11 NHW litters in total). The original LW litters were treated with a same program, so that there were 11 new low litter weight (NLW) litters as well. The latencies to first fighting, fighting frequencies and duration within three hours were recorded after regrouping and the lesions on body surface within 48 hours were scored. Besides, HR (heart rate, bpm, beats per minute) and activity count (ACT), time domain indexes and frequency domain indexes of the piglets were measured in an isolation trial to analyze the discrepancy in coping with stress between the original HW and LW litters. Results: The results exhibited that piglets from the HW litters launched fighting sooner and got statistically higher skin lesion score than those from the LW litters (p = 0.03 and 0.02, respectively). Regarding the HRV detection, compared with the HW litters, the LW litters exhibited a lower mean HR (p<0.05). In the isolation test, a highly significant higher ACT value was observed between the HW litters, compared to the LW litters (p<0.01). Significant differences were observed in standard deviation of R-R intervals, standard deviation of all normal to normal intervals, and most frequency-domain indicators: very low-frequency, low-frequency, and high frequency between the HW and LW litters as well. The difference in LF:HF was not significant (p = 0.779). Conclusion: This study suggests that compared with litters of low 20-day litter weights, litters with higher 20-day litter weight take more positive strategies to cope with stress and have stronger HRV regulation capacity; HW litters demonstrate better anti-stress and adaptation capacity in the case of regrouping and isolation.

Cytosolic prion protein induces apoptosis in human neuronal cell SH-SY5Y via mitochondrial disruption pathway

  • Wang, Xin;Dong, Chen-Fang;Shi, Qi;Shi, Song;Wang, Gui-Rong;Lei, Yan-Jun;Xu, Kun;An, Run;Chen, Jian-Ming;Jiang, Hui-Ying;Tian, Chan;Gao, Chen;Zhao, Yu-Jun;Han, Jun;Dong, Xiao-Ping
    • BMB Reports
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    • v.42 no.7
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    • pp.444-449
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    • 2009
  • Different neurodegenerative disorders like prion disease, is caused by protein misfolding conformers. Reverse-transfected cytosolic prion protein (PrP) and PrP expressed in the cytosol have been shown to be neurotoxic. To investigate the possible mechanism of neurotoxicity due to accumulation of PrP in cytosol, a PrP mutant lacking the signal and GPI (CytoPrP) was introduced into the SH-SY5Y cell. MTT and trypan blue assays indicated that the viability of cells expressing CytoPrP was remarkably reduced after treatment of MG-132. Obvious apoptosis phenomena were detected in the cells accumulated with CytoPrP, including loss of mitochondrial transmembrane potential, increase of caspase-3 activity, more annexin V/PI-double positive-stained cells and reduced Bcl-2 level. Moreover, DNA fragmentation and TUNEL assays also revealed clear evidences of late apoptosis in the cells accumulated CytoPrP. These data suggest that the accumulation of CytoPrP in cytoplasm may trigger cell apoptosis, in which mitochondrial relative apoptosis pathway seems to play critical role.

K-Ras-Activated Cells Can Develop into Lung Tumors When Runx3-Mediated Tumor Suppressor Pathways Are Abrogated

  • Lee, You-Soub;Lee, Ja-Yeol;Song, Soo-Hyun;Kim, Da-Mi;Lee, Jung-Won;Chi, Xin-Zi;Ito, Yoshiaki;Bae, Suk-Chul
    • Molecules and Cells
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    • v.43 no.10
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    • pp.889-897
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    • 2020
  • K-RAS is frequently mutated in human lung adenocarcinomas (ADCs), and the p53 pathway plays a central role in cellular defense against oncogenic K-RAS mutation. However, in mouse lung cancer models, oncogenic K-Ras mutation alone can induce ADCs without p53 mutation, and loss of p53 does not have a significant impact on early K-Ras-induced lung tumorigenesis. These results raise the question of how K-Ras-activated cells evade oncogene surveillance mechanisms and develop into lung ADCs. RUNX3 plays a key role at the restriction (R)-point, which governs multiple tumor suppressor pathways including the p14ARF-p53 pathway. In this study, we found that K-Ras activation in a very limited number of cells, alone or in combination with p53 inactivation, failed to induce any pathologic lesions for up to 1 year. By contrast, when Runx3 was inactivated and K-Ras was activated by the same targeting method, lung ADCs and other tumors were rapidly induced. In a urethane-induced mouse lung tumor model that recapitulates the features of K-RAS-driven human lung tumors, Runx3 was inactivated in both adenomas (ADs) and ADCs, whereas K-Ras was activated only in ADCs. Together, these results demonstrate that the R-point-associated oncogene surveillance mechanism is abrogated by Runx3 inactivation in AD cells and these cells cannot defend against K-Ras activation, resulting in the transition from AD to ADC. Therefore, K-Ras-activated lung epithelial cells do not evade oncogene surveillance mechanisms; instead, they are selected if they occur in AD cells in which Runx3 has been inactivated.

Comparison of the Efficacy of Different Organic Acids on Growth Performance and Nutrient Digestibility in Weaned Pigs (다양한 유기산제들의 급여가 이유자돈의 성장과 영양소 소화율에 미치는 효과)

  • Joo, Ji-Whan;Yang, YuXin;Choi, Jae-Yong;Choi, Soon-Chon;Cho, Won-Tak;Chae, Byung-Jo
    • Journal of Animal Science and Technology
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    • v.51 no.1
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    • pp.15-24
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    • 2009
  • Two experiments were conducted to compare the effects of different organic acids on growth performance and apparent nutrients digestibility in weaned pigs. In both the experiments, 180 pigs were assigned to four treatments with three replicates comprising of 15 pigs in each. Formic acid, ammonium-formate, lactic acid, and acid mixture were added to diets at 0.50% (Exp. 1) and 0.30% (Exp. 2) as dietary treatments for 5 and 6 wk feeding trial, respectively. The acid mixture was prepared by mixing formic acid and lactic acid at 50:50 ratios. To investigate the apparent ileal amino acids digestibility, twelve pigs (3 per treatment) were used and fitted with simple ileo-caecal T-cannula for both experiments. In Exp.1, growth performance was comparable (P>0.05) among pigs fed different organic acids, while acid mixture had higher (P<0.05) weight gain than that of lactic acid in Exp.2. The apparent ileal digestibility of amino acids was highest (P<0.05) in pigs fed acid mixture and lowest (P<0.05) in pigs fed formic acid diets in both experiments. These results indicated that supplementation with acid mixture (formic acid and lactic acid) improved performance and ileal amino acid digestibility in weaned pigs.

Angiostrongylus cantonensis: Scanning Electron Microscopic Observations on The Cuticle of Moulting Larvae

  • Zeng, Xin;Wei, Jie;Wang, Juan;Wu, Feng;Fung, Feng;Wu, Xiaoying;Sun, Xi;Zheng, Huanqing;Lv, Zhiyue;Wu, Zhongdao
    • Parasites, Hosts and Diseases
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    • v.51 no.6
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    • pp.633-636
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    • 2013
  • Angiostrongylus cantonensis is a parasitic nematode that needs to develop in different hosts in different larval stages. Freshwater snails, such as Pomacea canaliculata, are the intermediate host, and rats are the definitive host. Periodic shedding of the cuticle (moulting) is an important biological process for the survival and development of the parasite in the intermediate and definitive hosts. However, there are few studies on the cuticle alterations between different stages of this parasite. In this study, we observed the ultrastructural appearance and changes of the cuticle of the 2nd/3rd stage larvae (L2/L3) and the 3rd/4th stage larvae (L3/L4) using a scanning electron microscope. We also first divided L2/L3 into late L2 and early L3. The late L2 lacked alae, but possessed a pull-chain-like fissure. Irregular alignment of spherical particles on the cuticle were noted compared to the L3. Alae appeared in the early L3. The old cuticle turned into a thin filmlike structure which adhered to the new cuticle, and spherical particles were seen regularly arranged on the surface of this structure. Regular rectangular cavities were found on the surface of L3/L4. The caudal structure of L3/L4 was much larger than that of L3, but caudal inflation, such as seen in L4, was not observed. These results are the first to reveal the ultrastructural changes of the cuticle of A. cantonensis before and after moulting of L2/L3 and L3/L4.

Evaluation of Dry Matter Intake and Average Daily Gain Predicted by the Cornell Net Carbohydrate and Protein System in Crossbred Growing Bulls Kept in a Traditionally Confined Feeding System in China

  • Du, Jinping;Liang, Yi;Xin, Hangshu;Xue, Feng;Zhao, Jinshi;Ren, Liping;Meng, Qingxiang
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.11
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    • pp.1445-1454
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    • 2010
  • Two separate animal trials were conducted to evaluate the coincidence of dry matter intake (DMI) and average daily gain (ADG) predicted by the Cornell Net Carbohydrate and Protein System (CNCPS) and observed actually in crossbred growing bulls kept in a traditionally confined feeding system in China. In Trial 1, 45 growing Simmental${\times}$Mongolia crossbred F1 bulls were assigned to three treatments (T1-3) with 15 animals in each treatment. Trial 2 was conducted with 60 Limousin${\times}$Fuzhou crossbred F2 bulls allocated to 4 treatments (t1-4). All of the animals were confined in individual stalls. DMI and ADG for each bull were measured as a mean of each treatment. All of the data about animals, environment, management and feeds required by the CNCPS model were collected, and model predictions were generated for animals on each treatment. Subsequently, model-predicted DMI and ADG were compared with the actually recorded results. In the three treatments in Trial 1, 93.3, 80.0 and 73.3% of points fell within the range from -0.4 to 0.4 kg/d for DMI mean bias; similarly, in the four treatments in Trial 2, about 86.7, 73.3, 73.3 and 80.0% of points fell within the same range. These results indicate that the CNCPS model can accurately predict DMI of crossbred bulls in the traditionally confined feeding system in China. There were no significant differences between predicted and observed ADG for T1 (p = 0.06) and T2 (p = 0.09) in Trial 1, and for t1 (p = 0.07), t2 (p = 0.14) and t4 (p = 0.83) in Trial 2. However, significant differences between predicted and observed ADG values were observed for T3 in Trial 1 (p<0.01) and for t3 in Trial 2 (p = 0.04). By regression analysis, a statistically different value of intercept from zero for the regression equation of DMI (p<0.01) or an identical value of ADG (p = 0.06) were obtained, whereas the slopes were significantly different (p<0.01) from unity for both DMI and ADG. Additionally, small root mean square error (RMSE) values were obtained for the unbiased estimator of the two variances (DMI and ADG). Thus, the present results indicated that the CNCPS model can give acceptable estimates of DMI and ADG of crossbred growing bulls kept in a traditionally confined feeding system in China.