• Title/Summary/Keyword: in-vitro

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Corn-Based Forage Cropping Systems in Rice Black-Streaked Dwarf Virus Prevalent Area (흑조위축병이 심한 남부지방에서 옥수수를 중심으로 한 사료작물 작부체계)

  • 이석순;이진모
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.34 no.1
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    • pp.30-39
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    • 1989
  • Forage productivity of cropping systems of rye - silage corn, silage corn - oats, silage corn - rape was studied in the south-eastern part of Korea where rice black-streaked dwarf virus(RBSDV) infection of corn are severe. Rye(cv. Paldanghomil) was planted on Oct. 20 of 1986 and harvested 10 times from April 5 to May 5 at the 5-day intervals in 1987, corn (cv. Suweon 19 and Jinjuok) was planted 5 times from April 5 to May 15 at the 10-day intervals in 1987, and oats(cv. Megwiri) and rape (cv. Velox) were planted 4 times from Sept: 4 to 25 at the 7-day intervals and harvested 4 times from Nov. 10 to Dec. 10 at the 10-day intervals in 1987. Considering yield, nutrition value, and in vitro dry matter digestibility (IVDMD), forage productivity of the cropping systems was compared. As harvesting time of rye delayed, plant height, dry matter(DM) yield, percent DM, crude fiber, and digestible DM yield increased, but crude protein, crude fat, and IVDMD decreased. However, nitrogen free extract was not different among the harvesting dates. As planting date of corn delayed, RBSDV infection rate increased. but DM yield of silage decreased. However, silage yield of Jinjuok was higher, but RBSDV infection rate was lower compared with Suweon 19 at all planting dates. DM yield of oats and rape decreased as planting date delayed. However, at Sept. 4 and 11 plantings yield of oats on Nov. 10 was much lower than that of rape, but the differences in yield between two crops decreased with delayed harvesting, and yield was similar on Dec. 10. A cropping system harvesting rye around April 20 and followed by planting corn in late April was best among the rye-corn systems considering yield and nutrition value of both crops. However, among the corn-oats or corn-rape cropping systems early April planting of corn and followed by early Sept. planting of oats or rape showed best results with similar yield potential of the best rye-corn cropping system.

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A Study on Anti-Bone Resorption & Osteoporosis by Taeyoungion-Jahage Extracts

  • Bae, Hyo-Sang;Cho, Hyung-Lae;Kim, Dong-Il;Lee, Tae-Kyun;Kim, Jun-Ki;Shin, Jung-Sik
    • The Journal of Korean Obstetrics and Gynecology
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    • v.15 no.4
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    • pp.61-75
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    • 2002
  • Recombinant human $interleukin-1{\beta}$ $(rhIL-1{\beta})$ regulates several activities of the osteoblast cells derived from mouse calvarial bone explants in vitro. $rhIL-1{\beta}$ stimulated cellular proliferation and the synthesis of prostaglandin $E_2(PGE_2)$ and plasminogen activator activity in the cultured cells in a dose-dependent manner. However, the induction of osteocalcin synthesis and alkaine phosphatase activity in response to vitamine D, two characteristics of the osteoblast phenotype, were antagonized by $rhIL-1{\beta}$ over a similar dose range. This study supports the role of $IL-1{\beta}$ in the pathological modulation of bone cell metabolism, with regard to implication in the pathogenesis of osteoporosis by $IL-1{\beta}$. When the mouse calvarial bone cells were used, the bone resorption induced by $IL-1{\beta}$ was strongly inhibited by calcitonin treatment, indicating osteoclast-mediated bone resorption. On the other hand, the medicinal extracts of Taeyoungjon-Jahage (T.Y.J-J.H.G extracts) was tested for whether they could inhibit $IL-1{\beta}-induced$ $PGE_2$ production. Cell viability was not significantly affected by treatment with the indicated concentration of the extracts. The T.Y.J.-J.H.G. extracts were shown to have the inhibitory effects against the synthesis of $PGE_2$. We also examined the effect of the pretreatment with a various concentrations of the T.Y.J.-J.H.G. extracts then treated the $PGE_2-induction$ agents. Pretreatment of the T.Y.J.-J.H.G. extracts for 1 h, which by itself had little effect on cell survival, did not enhance the synthesis of $PGE_2$. Furthermore, the T.Y.J-J.H.G. extracts were shown to have the protective effects against plasminogen dependent fibrinolysis induced by the bone resorption agents of $IL-1{\beta}$. Pretreatment of the T.Y.J.-J.H.G. extracts for 1 h did not enhance the plasminogen dependent fibrinolysis. Finally, calcitonin showed the inhibitory activity the $IL-1{\beta}-stimulated$ bone resorption in the mouse calvarial bone cells having both of the osteoblast and osteoclast cells. Seemingly, pretreatment of the T.Y.J.-J.H.G. extracts for 1 h reduced the bone resorption. These results clearly indicated that calcitonin and T.Y.J.-J.H.G. extracts play key roles in inhibition of the osteoclast-mediated bone resorption.

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EFFECT OF LIGHT IRRADIATION MODES ON THE MARGINAL LEAKAGE OF COMPOSITE RESIN RESTORATION (광조사 방식이 복합레진 수복물의 변연누출에 미치는 영향)

  • 박은숙;김기옥;김성교
    • Restorative Dentistry and Endodontics
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    • v.26 no.4
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    • pp.263-272
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    • 2001
  • The aim of this study was to investigate the influence of four different light curing modes on the marginal leakage of Class V composite resin restoration. Eighty extracted human premolars were used. Wedge-shaped class Y cavities were prepared on the buccal surface of the tooth with high-speed diamond bur without bevel. The cavities were positioned half of the cavity above and half beyond the cemento-enamel junction. The depth, height, and width of the cavity were 2 mm, 3 mm and 2 mm respectively. The specimens were divided into 4 groups of 20 teeth each. All the specimen cavities were treated with Prime & Bond$^{R}$ NT dental adhesive system (Dentsply DeTrey GmbH, Germany) according to the manufacturer's instructions and cured for 10 seconds except group VI which were cured for 3 seconds. All the cavities were restored with resin composite Spectrum$^{TM}$ TPH A2 (Dentsply DeTrey GmbH, Germany) in a bulk. Resin composites were light-cured under 4 different modes. A regular intensity group (600 mW/${cm}^2$, group I) was irradiated for 30 s, a low intensity group (300 mW/${cm}^2$, group II) for 60 s and a ultra-high intensity group (1930 mW/${cm}^2$, group IV) for 3 s. A pulse-delay group (group III) was irradiated with 400 mW/${cm}^2$ for 2 s followed by 800 mW/${cm}^2$ for 10 s after 5 minutes delay. The Spectrum$^{TM}$ 800 (Dentsply DeTrey GmbH, Germany) light-curing units were used for groups I, II and III and Apollo 95E (DMD, U.S.A.) was used for group IV. The composite resin specimens were finished and polished immediately after light curing except group III which were finished and polished during delaying time. Specimens were stored in a physiologic saline solution at 37$^{\circ}C$ for 24 hours. After thermocycling (500$\times$, 5-55$^{\circ}C$), all teeth were covered with nail varnish up to 0.5 mm from the margins of the restorations, immersed in 37$^{\circ}C$, 2% methylene blue solution for 24 hours, and rinsed with tap water for 24 hours. After embedding in clear resin, the specimens were sectioned with a water-cooled diamond saw (Isomet$^{TM}$, Buehler Co., Lake Bluff, IL, U.S.A.) along the longitudinal axis of the tooth so as to pass the center of the restorations. The cut surfaces were examined under a stereomicroscope (SZ-PT Olympus, Japan) at ${\times}$25 magnification, and the images were captured with a CCD camera (GP-KR222, Panasonic, Japan) and stored in a computer with Studio Grabber program. Dye penetration depth at the restoration/dentin and the restoration/enamel interfaces was measured as a rate of the entire depth of the restoration using a software (Scion image, Scion Corp., U.S.A.) The data were analysed statistically using One-way ANOVA and Tukey's method. The results were as follows : 1. Pulse-Delay group did not show any significant difference in dye penetration rate from other groups at enamel and dentin margins (p>0.05) 2. At dentin margin, ultra-high intensity group showed significantly higher dye penetration rate than both regular intensity group and low intensity group (p<0.05). 3. At enamel margin, there were no statistically significant difference among four groups (p>0.05). 4. Dentin margin showed significantly higher dye penetration rate than enamel margin in all groups (p<0.05).

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In vitro cytotoxicity of self-etching primers (자가 산부식 프라이머의 세포독성에 관한 실험적 연구)

  • Rhee, Chang-Hoon;Kim, In-Ryeon;Kim, Gyoo-Cheon;Kim, Seong-Sik;Son, Woo-Sung
    • The korean journal of orthodontics
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    • v.36 no.6
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    • pp.422-433
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    • 2006
  • Objective: Several ions and components are released from self-etching primers in the oral cavity. This may cause injury to the periodontal tissues throughout orthodontic treatment. The purpose of this study was to assess the cytotoxicity of self-etching primers to HGF-1, HaCaT, and RHEK cells. Method: Transbond XT Primer (3M Unitek, Monrovia, CA, USA), and self-etching primers, Clearfil SE Bond (Kuraray, Osaka, Japan), Transbond Plus SEP (3M Unitek, Monrovia, CA, USA), and Adper Prompt L-Pop (3M Unitek, Monrovia, CA, USA), were evaluated by MTT assay, and cellular changes were also observed. Results: In all cells after 72 hours with all primers, severe morphological changes such as atrophy and necrosis were observed. In the MTT assay using HGF-1, Clearfil SE Bond, Transbond XT Primer, Transbond Plus SEP, and Adper Prompt L-Pop were lined up in order of ascending cytotoxicity When using HaCaT, Clearfil SE Bond, Adper Prompt L-Pop, Transbond Plus SEP, and Transbond XT Primer were lined up in order of ascending cytotoxicity. When using RHEK, Clearfil SE Bond, Transbond XT Primer, Adper Prompt L-Pop, and Transbond Plus SEP were lined up in order of ascending cytotoxicity. Conclusion: The result of this study shows that care is needed because self-etching primers show cytotoxic properties similar to conventional primers.

Adhesion-induced generation of oxygen free radical from human alveolar macrophages and its mechanisms (폐포대식세포의 부착에 의한 산소유리기 분비능 활성화 및 그 기전)

  • Chung, Man-Pyo;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.2
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    • pp.210-220
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    • 1996
  • Background : Neutrophils or monocytes separated in vitro by the adherence to plastic surface are known to be activated by surface adherence itself and subsequent experimental data might be altered by surface adherence. In the process of surface adherence, adhesion molecules have a clear role in intracellular signal pathway of cellular activation. Human alveolar macrophages(HAM) are frequently purified by the adherence procedure after bronchoalveolar lavage. But the experimental data of many reports about alveolar macrophages have ignored the possibility of adhesion-induced cellular activation. Method : Bronchoalveolar lavage was performed in the person whose lung of either side was confirmed to be normal by chest CT. With the measurement of hydrogen peroxide release from adherent HAM to plastic surface and non-adherent HAM with or without additional stimulation of phorbol myristate acetate(PMA) or N-formyl-methionyl-leucyl-phenylalanine (fMLP), we observed the effect of the adherence to plastic surface. We also evaluated the effect of various biological surfaces on adhesion-induced activation of HAM. Then, to define the intracellular pathway of signal transduction, pretreatment with cycloheximide, pertussis toxin and anti-CD11/CD18 monoclonal antibody was done and we measured hydrogen peroxide in the culture supernatant of HAM. Results : 1) The adherence itself to plastic surface directly stimulated hydrogen peroxide release from human alveolar macrophages and chemical stimuli such as phorbol myristate acetate(PMA) or N-formyl-methionyl-leucyl-phenylalanine(fMLP) colud not increase hydrogen peroxide release in these adherent macrophages which is already activated. 2) PMA activated human alveolar macrophages irrespective of the state of adhesion. However, fMLP stimulated the release of hydrogen peroxide from the adherent macrophages, but not from the non-adherent macrophages. 3) HAM adherent to A549 cell(type II alveolar epithelium-like human cell line) monolayer released more hydrogen peroxide in response to both PMA and fMLP. This adherence-dependent effect of fMLP was blocked by pretreatment of macrophages with cycloheximide, pertussis toxin and anti-CD18 monoclonal antibody, Conclusion : These results suggest that the stimulatory effect of PMA and fMLP can not be found in adherent macrophage because of the activation of human alveolar macrophage by the adherence to plastic surface and the cells adhered to biologic surface such as alveolar epithelial cells are appropriately responsive to these stimuli. It is also likely that the effect of fMLP on the adherent macrophage requires new protein synthesis via G protein pathway and is dependent on the adhesion between alveolar macrophages and alveolar epithelial cells by virtue of CD11/CD18 adhesion molecules.

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IN VITRO COMPARISON OF VARIOUS DIAGNOSTIC METHODS OF OCCLUSAL CAR10US LESIONS (교합면 우식병소의 다양한 진단법에 관한 비교연구)

  • Kim, Jae-Gon;Kim, Young-Jin;Kim, Young-Sin;Baik, Byeong-Ju
    • Journal of the korean academy of Pediatric Dentistry
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    • v.28 no.4
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    • pp.613-619
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    • 2001
  • The aims of this study were to compare the accuracy, sensitivity and specificity of cnventional visual examination, radiography and a new laser fluorescence method, KaVo Diagnodent, for the detection of occlusal caries lesions. One hundred sound human premolars and molars which had no restorations or interproximal cavities were tested by three methods. Tooth lesions depth was assessed at histologic examination using Caries detector dye The following results were obtained. 1. Diagnodent show 7.8 in sound tooth, 25.4 in initial caries, 30.5 in enamel caries, and 53.8 in dentin caries with average score 2. Spearman and Pearson relation coefficient was high between tooth-specimen test with dye and Diagnodent(0.736, 0.619), visual examination(0.664, 0.666), and was low between tooth-specimen test with dye and radiographic examination(P<0.01, total) 3. Accuracy of occlusal caries was highest on Diagnodent(65%) and lowest on radiographic examination(35%) 4. In initial caries, the sensitivity and specificity of Diagnodent method was the highest. In enamel caries, the sensitivity of visual examination was the highest and specificity of Diagnodent method was the highest. In dentinal caries, the sensitivity and specificity of Diagnodent method was the highest and sensitivity of visual examination was the lowest.

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Role of HOXA Gene in Human Endometrial Decidualization (인간 자궁내막의 탈락막화에서 HOXA10 유전자의 역할)

  • Lee, Chang-Se;Park, Dong-Wook;Park, Chan-Woo;Kim, Tae-Jin
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.3
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    • pp.207-216
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    • 2010
  • Objective: This study was performed to clarify the role of HomeoboxA (HOXA) and its related signaling molecules in the decidualization of primary cultured endometrial cells. Methods: Human endometrial tissues were obtained by curettage of hysterectomy specimens from patients with conditions other than endometrial diseases. Tissues were minced and digested with Trypsin-EDTA for 20 min, $37^{\circ}C$. Cells were cultured with DMEM/F12 medium in $37^{\circ}C$, 5% $CO_2$ incubator for 24 hrs. Cells were treated with HOXA10 siRNA and added transforming growth factor (TGF)-${\beta}1$ (10 ng/mL) for 48 hrs to induces decidualization in vitro. Reverse transcription polymerase chain reaction analysis was accomplished to observe the expression of HOXA10, prolactin, cyclooxygenase (COX)-2, peroxisome proliferatoractivated receptor (PPAR)-$\gamma$, and wingless-type MMTV integration site family (Wnt). Results: HOXA10 expression was increased (1.8 fold vs. non-treated control) in TGF-${\beta}1$ treated cells. Decidualization marker, prolactin, was significantly increased in TGF-${\beta}1$ treated cells compared with HOXA10 siRNA treated cells. Endometrial cell differentiation marker, COX-2 was down-regulated by HOXA10 siRNA even if cells were treated with TGF-${\beta}1$. Wnt4 was down-regulated by treated with HOXA10 siRNA, this expression patters was not changed by TGF-${\beta}1$. Expression of PPAR-$\gamma$ was down regulated by TGF-${\beta}1$ in regardless of HOXA10 siRNA treatment. Conclusion: TGF-${\beta}1$ which is induced by progesterone in endometrial epithelial cells may induces stromal cell decidualization via HOXA10 and Wnt signaling cascade.

Phospholipase C-γ Activation by Direct Interaction with β-Tubulin Isotypes (베타 튜불린에 의한 포스포리파제 C-감마1의 활성화)

  • Lee, In-Bum;Kim, Sung-Kuk;Choi, Jang-Hyun;Suh, Pann-Ghill;Chang, Jong-Soo
    • Journal of Life Science
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    • v.16 no.4
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    • pp.612-617
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    • 2006
  • Phosphoinositide-specific phospholipase $C-{\gamma}\;1\; (PLC-{\gamma}\;1)$ has pivotal roles in cellular signaling by producing second messengers, inositol 1,4,5-trisphosphate $(IP_3)$ and diacylglycerol (DG). Tubulin is a main component of microtubules and mitotic spindle fibers, which are composed of ${\alpha}-$ and ${\beta}-tubulin$ heterodimers in all eukaryotic cells. In humans, six ${\beta}-tubulin$ isotypes have been identified which display a distinct pattern of tissue expression. Previously we found that $PLC-{\gamma}\;1$ and one of four ${\beta}-tubulin$ isotypes including ${\beta}1$, ${\beta}2$, ${\beta}3$ and ${\beta}6$, colocalized in COS-7 cells and cotranslocated to the plasma membrane to activate $PLC-{\gamma}\;1$ upon agonist stimulation. In the present study, we demonstrate that the remaining two, tubulin ${\beta}4$ and ${\beta}5$, also showed a potential to activate $PLC-{\gamma}\;1$. The phosphatidylinositol 4,5-bisphosphate $(PIP_2)$ hydrolyzing activity of $PLC-{\gamma}\;1$ was substantially increased in the presence of purified ${\beta}4$ and ${\beta}5$ tubulin in vitro, whereas the activity was not promoted by bovine serum albumin, suggesting that tubulin ${\beta}4$ and ${\beta}5$ also activate $PLC-{\gamma}\;1$. Taken together, our results suggest that all the ${\beta}-tubulin$ isotype activates $PLC-{\gamma}\;1$ activity to regulate cellular signaling.

Transcriptome Analyses for the Anti-Adipogenic Mechanism of an Herbal Composition (생약복합물의 지방세포형성억제 기전규명을 위한 전사체 분석)

  • Lee, Hae-Yong;Kang, Ryun-Hwa;Bae, Sung-Min;Chae, Soo-Ahn;Lee, Jung-Ju;Oh, Dong-Jin;Park, Suk-Won;Cho, Soo-Hyun;Shim, Yae-Jie;Yoon, Yoo-Sik
    • Journal of Life Science
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    • v.20 no.7
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    • pp.1054-1065
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    • 2010
  • SH21B is a natural composition composed of seven herbs: Scutellaria baicalensis Georgi, Prunus armeniaca Maxim, Ephedra sinica Stapf, Acorus gramineus Soland, Typha orientalis Presl, Polygala tenuifolia Willd and Nelumbo nucifera Gaertner (Ratio 3:3:3:3:3:2:2). In our previous study, we reported that SH21B inhibited adipogenesis and fat accumulation in 3T3-L1 cells through modulation of various regulators in the adipogenesis pathway. The aim of this study was to analyze the transcriptome profiles for the anti-adipogenic effects of SH21B in 3T3-L1 cells. Total RNAs from SH21B-treated 3T3-L1 cells were reverse-transcribed into cDNAs and hybridized to Affymetrix Mouse Gene 1.0 ST array. From microarray analyses, we identified 2,568 genes of which expressions were changed more than two-fold by SH21B, and the clustering analyses of these genes resulted in 9 clusters. Three clusters among the 9 showed down-regulation by SH21B (cluster 4, cluster 6 and cluster 9), and two clusters showed up-regulation by SH21B (cluster 7 and cluster 8) during the adipogenesis of 3T3-L1 cells. It was found that many genes related to cell proliferation and adipogenesis were included in these clusters. Clusters 4, 6 and 9 included genes which were related with adipogenesis induction and cell cycle arrest. Clusters 7 and 8 included genes related to cell proliferation as well as adipogenesis inhibition. These results suggest that the mechanisms of the anti-adipogenic effects of SH21B may be the modulation of genes involved in cell proliferation and adipogenesis.

Onion Beverages Improve Amyloid β Peptide-Induced Cognitive Defects via Up-Regulation of Cholinergic Activity and Neuroprotection (양파(Allium cepa L.) 음료의 콜린성 활성 증가 및 뇌신경세포 보호로 인한 Amyloid β Peptide 유도에 대한 인지장애 개선 효과)

  • Park, Seon Kyeong;Kim, Jong Min;Kang, Jin Yong;Ha, Jeong Su;Lee, Du Sang;Kim, Ah-Na;Choi, Sung-Gil;Lee, Uk;Heo, Ho Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.11
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    • pp.1552-1563
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    • 2016
  • To examine the cognitive function of onion (Allium cepa L.) beverages (odourless and fortified), we analyzed in vitro neuronal cell protection against $H_2O_2$-induced cytotoxicity and performed in vivo tests on amyloid beta ($A{\beta}$)-induced cognitive dysfunction. Cellular oxidative stress and cell viability were evaluated by DCF-DA assay and MTT assay. These results show that fortified beverage resulted in better neuronal cell protection than odourless beverage at lower concentration ($0{\sim}100{\mu}g/mL$). Fortified beverage also showed more excellent acetylcholinesterase (AChE) inhibitory activity ($IC_{50}$: 4.20 mg/mL) than odourless beverage. The cognitive functions of odourless beverage and fortified beverage in $A{\beta}$-induced neurotoxicity were assessed by Y-maze, passive avoidance, and Morris water maze tests. The results show improved cognitive function in both groups treated with beverages. After in vivo tests, cholinergic activities were determined based on AChE inhibition and acetylcholine levels, and antioxidant activities were measured as SOD, oxidized glutathione (GSH)/total GSH ratio, and MDA levels in mouse brain tissue. In a Q-TOF UPLC/MS system, main compounds were analyzed as follows: odourless beverage (five types of sugars and three types of phenolics) and fortified beverages (six types of phenolics and two types of steroidal saponins).