• Title/Summary/Keyword: In vitro evaluation

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Radiation-Induced Apoptosis of Lymphocytes in Peripheral Blood (말초혈액 내 림프구의 방사선에 의한 아포프토시스)

  • Oh, Yoon-Kyeong;Lee, Tae-Bum;Nam, Taek-Keun;Kee, Keun-Hong;Choi, Cheol-Hee
    • Radiation Oncology Journal
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    • v.21 no.1
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    • pp.75-81
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    • 2003
  • Purpose : This study quantitatively evaluated the apoptosis In human peripheral blood lymphocytes using flow cytometry, and investigated the possibility of using this method, with a small amount of blood, and the time and dose dependence of radiation-induced apoptosls. Materials and Methods : Peripheral blood lymphocyes were isolated from the heparinized venous blood of 11 healthy volunteers, 8 men and 3 women, with each 10 ml of blood being divided Into IS samples. The blood lymphocytes were Irradiated using a linear accelerator at a dose rate of 2.4 Gy/min, to deliver doses of 0.5, 1, 2 and S Gy. The control samples, and Irradiated cells, were maintained in culture medium for 24, 48 and 72 hours fellowing the Irradiation. The number of apoptotic cells after the in vitro X-irradiation was measured by flow cytometry after Incubation periods of 24, 48 and 72 hours. We also observed the apoptotic cells using a DNA fragmentation assay and electron microscopy. Results : The rate oi spontaneous apoptosis increased in relation to the time interval following irradiation (1.761 ${\pm}$0.161, 3.563${\pm}$0.554, 11.098${\pm}$2.849, at 24, 48, and 72 hours). The apoptotli cells also increased In the samples irradiated with 0.5, 1, 2 and 5 Gy, In a radiation dose and time interval after Irradiation manner, with the apoptosls being too great at 72 hours after Irradiation. The dose-response curves were characterized by an Initial steep Increase In the number of apoptotic cells for Irradiation doses below 2 Gy, with a flattening of the curves as the dose approached towards 5 Gy. Conclusion :The flow cytometric assay technique yielded adequate data, and required less than 1 mL of blood. The time and dose dependence of the radiation-induced apoptosis, was also shown. It is suggested that the adequate time Interval required for the evaluation of apoptosis would be 24 to 48 hours after blood sampling.

Evaluation on the Usefulness of Filter in Sentinel Lymphoscintigraphy Using $^{99m}Tc$-Phytate (Phytate를 이용한 감시림프절 검사 시 Filter의 유용성 평가)

  • Jeong, Yeong-Hwan;Seo, Han-Kyung;Shim, Cheol-Min;Lim, Seong-Dong;Han, Dong-Hyeon;Park, Yung-Sun;Kim, Dong-Yun
    • The Korean Journal of Nuclear Medicine Technology
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    • v.14 no.1
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    • pp.35-39
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    • 2010
  • Purpose: The aim of this study was to investigate distribution of particle size in phytate kit and compare filtered method with non-filtered method using 200 nm filter for sentinel lymphoscintigraphy (SLS). Materials and Methods: Five phytate kit of having the same available period was measured by particle size analyzer. For in-vivo experiment, $^{99m}Tc$-phytate was injected intradermally at both foot to perform lymphoscintigraphy. Imaging was acquired at 1hour after injection. Region of interest (ROI) was drawn in inguinal and background area for analysis. RAW 264.7 cells (Murine macrophage cell) were prepared for measurement of celluar uptake as a representative of macrophages. Paired t-test was performed using SPSS (SPSS Inc, USA) for statistical analysis. Results: The size of most particle in Techne phytate kit was distributed in 130~650 nm(90.5 %). In-vivo study, the ROI analysis showed similar result between filtered and non-filtered sample, and the numerical value of count/pixel were $58.3{\pm}5.97$ and $60.2{\pm}4.88$. In-vitro study, cellular uptake study also showed no difference between filtered and non-filtered sample by gamma counting. Conclusion: The present study demonstrates that there was no meaning of 200 nm filtered method for SLS using $^{99m}Tc$-phytate.

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Development of High Intensity Focused Ultrasound (HIFU) Mediated AuNP-liposomal Nanomedicine and Evaluation with PET Imaging

  • Ji Yoon Kim;Un Chul Shin;Ji Yong Park;Ran Ji Yoo;Soeku Bae;Tae Hyeon Choi;Kyuwan Kim;Young Chan Ann;Jin Sil Kim;Yu Jin Shin;Hokyu Lee;Yong Jin Lee;Kyo Chul Lee;Suhng Wook Kim;Yun-Sang Lee
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.9 no.1
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    • pp.9-16
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    • 2023
  • Liposomes as drug delivery system have proved useful carrier for various disease, including cancer. In addition, perfluorocarbon cored microbubbles are utilized in conjunction with high-intensity focused-ultrasound (HIFU) to enable simultaneous diagnosis and treatment. However, microbubbles generally exhibit lower drug loading efficiency, so the need for the development of a novel liposome-based drug delivery material that can efficiently load and deliver drugs to targeted areas via HIFU. This study aims to develop a liposome-based drug delivery material by introducing a substance that can burst liposomes using ultrasound energy and confirm the ability to target tumors using PET imaging. Liposomes (Lipo-DOX, Lipo-DOX-Au, Lipo-DOX-Au-RGD) were synthesized with gold nanoparticles using an avidin-biotin bond, and doxorubicin was mounted inside by pH gradient method. The size distribution was measured by DLS, and encapsulation efficiency of doxorubicin was analyzed by UV-vis spectrometer. The target specificity and cytotoxicity of liposomes were assessed in vitro by glioblastoma U87mg cells to HIFU treatment and analyzed using CCK-8 assay, and fluorescence microscopy at 6-hour intervals for up to 24 hours. For the in vivo study, U87mg model mouse were injected intravenously with 1.48 MBq of 64Cu-labeled Lipo-DOX-Au and Lipo-DOX-Au-RGD, and PET images were taken at 0, 2, 4, 8, and 24 hours. As a result, the size of liposomes was 108.3 ± 5.0 nm at Lipo-DOX-Au and 94.1 ± 12.2 nm at Lipo-DOX-Au-RGD, and it was observed that doxorubicin was mounted inside the liposome up to 52%. After 6 hours of HIFU treatment, the viability of U87mg cells treated with Lipo-DOX-Au decreased by around 20% compared to Lipo-DOX, and Lipo-DOX-Au-RGD had a higher uptake rate than Lipo-DOX. In vivo study using PET images, it was confirmed that 64Cu-Lipo-DOX-Au-RGD was taken up into the tumor immediately after injection and maintained for up to 4 hours. In this study, drugs released from liposomes-gold nanoparticles via ultrasound and RGD targeting were confirmed by non-invasive imaging. In cell-level experiments, HIFU treatment of gold nanoparticle-coupled liposomes significantly decreased tumor survival, while RGD-liposomes exhibited high tumor targeting and rapid release in vivo imaging. It is expected that the combination of these models with ultrasound is served as an effective drug delivery material with therapeutic outcomes.

Fermentation Properties and Increased Health Functionality of Kimchi by Kimchi Lactic Acid Bacteria Starters (김치 유산균 Starter를 이용한 김치의 발효 특성 및 기능성 증진 효과)

  • Bong, Yeon-Ju;Jeong, Ji-Kang;Park, Kun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.11
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    • pp.1717-1726
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    • 2013
  • Fermentation characteristics and health functionalities of kimchi by inoculating kimchi lactic acid bacteria (LAB) starters were studied. We manufactured single LAB starter kimchi (Lactobacillus plantarum pnuK, Lactobacillus plantarum 3099K, Leuconostoc mesenteroides pnuK), mixed LAB starter kimchi (Lb. plantarum pnu/Leu. mesenteroides pnuK, Lb. plantarum 3099/Leu. mesenteroides pnuK) with inoculum size of $10^6$ CFU/g, as well as naturally fermented kimchi (NK), and fermented them for 6 days at $15^{\circ}C$. The pH and acidity of the early phase of fermentation were not different, but kimchi with the starters showed rapid changes in the pH and acidity from 2 days of fermentation. As the fermentation progressed, the level of total aerobic bacteria and Lactobacillus sp. increased similarly with or without Lb. plantarum (LP) inoculation. However, the level of Leuconostoc sp. was high in kimchi inoculated with Leuconostoc sp. starter. In the sensory evaluation test, kimchi with starters received higher overall acceptability scores than those of NK; mixed starter added kimchi earned the highest score. In DPPH and hydroxyl radical scavenging activity, kimchi with the starters exhibited higher activity than that of NK. In the MTT assay of HCT-116 and HT-29 human colon cancer cells, NK showed inhibition rates of 63.4 and 51.9%, but LPpnuK achieved 77.1 and 68.8%, respectively. This study showed that inoculating starters in kimchi increased in vitro antioxidant and anticancer activities, and single starter (LP) added kimchi revealed higher functionality than the kimchi with mixed starter. Kimchis with the starters effectively up-regulated the gene expressions of the pro-apoptotic gene of Bax, but down-regulated Bcl-2. They promoted expressions of p53 and p21, and suppressed expressions of inflammation-related genes, iNOS and COX-2, compared with NK. Taken together, it is expected that using starters may help manufacture kimchi with improved sensory quality and health functionality.

Evaluation of time-dependent antimicrobial effect of sodium dichloroisocyanurate (NaDCC) on Enterococcus faecalis in the root canal (이염화이소시아뉼산나트륨 제재의 근관 내 사용 시 시간에 따른 E. faecalis에 대한 항균성 평가)

  • Kim, Hye-Jeong;Park, Se-Hee;Cho, Kyung-Mo;Kim, Jin-Woo
    • Restorative Dentistry and Endodontics
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    • v.32 no.2
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    • pp.121-129
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    • 2007
  • The purpose of this study was to assess the antibacterial effect of sodium dichloroisocyanurate (NaDCC), sodium hypochlorite (NaOCl), and chlorhexidine (CHX) on Enterococcus faecalis and to evaluate and to compare the time-dependant antimicrobial effect of NaDCC with NaOCl and CHX in the root canal in vitro before and after instrumentation. Extracted Human single teeth were prepared by serial instrumentation technique. The samples were autoclaved and contaminated for 3 days with E. faecalis monocultures. The teeth were then divided into 4 groups Each group was irrigated and inserted with 2% NaOCl, 2% NaDCC, 2% CHX and steri)ized saline. After 6, 12, 24, 72h, and 1 week incubation, sterilized paper point was inserted into the root canal. Paper points containing root canal contents were then placed on the agar plate. And then each root cana) was prepared with #4 and #5 GG (Gates-Glidden) drill. The debris were collected in the sterilized microtube and the plates were incubated at $37^{\circ}C$ in an increased $CO_2$ atmosphere. After 24h incubation the growth of bacteria around the paper points were measured. NaOCl and NaDCC solution shows similar antimicrobial effect for E. faecalis at 6, 12, 24, 72h and 1 week. In centrol group, irrigated with sterilized saline, no antimicrobial effect was observed. The results are in agreement with other investigators, who have shown the bactericidal property and possibility of NaDCC as a root canal irrigation solution. Thus it seems that NaDCC solutions can be clinically applied into the root canal within 1 week after dilution.

Evaluation of in-vitro Antithrombosis Activity of Lees of Korean Traditional Wine (전통주 주박의 항혈전 활성 평가)

  • Kim, Mi-Sun;Lee, Ye-Seul;Kim, Jong Sik;Shin, Woo-Chang;Sohn, Ho-Yong
    • Journal of Life Science
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    • v.24 no.8
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    • pp.865-872
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    • 2014
  • In this study, ethanol and hot water extracts of lees from Korean traditional wine (J-B, J-S, J-Y, J-H, and J-W) were prepared, and their effects on blood coagulation, platelet aggregation, and hemolysis of human red blood cells (hRBCs) were investigated to develop functional food ingredients from lees. The pH and brix of the lees ranged from 3.90 to 4.29 and 5.0 to $27.0^{\circ}$, respectively, and there was a huge difference in the water and ethanol content among the lees. The nuruk and additives used affected the color and physicochemical properties of lees. The J-W takju made from only rice and traditional nuruk, which has $13^{\circ}$ brix and 1.8% of alcohol, has potential as functional food ingredient. With regard to the extraction yields of lees, higher yields were obtained from J-H, which contains different medicinal plants, in ethanol, followed by J-W, J-B, J-S, and J-Y. Higher extraction yields of lees were obtained from J-S in hot water, followed by J-B, J-W, J-H, and J-Y, respectively. The ethanol extract of J-H and the hot water extract of J-Y had the highest contents of total polyphenol and total flavonoids among the lees extracts. The 10 lees extracts did not show hemolysis activity against hRBCs up to 5 mg/ml. In an anticoagulation activity assay, the ethanol extracts of three yakju lees (J-B, J-S, and J-Y) and the hot water extract of J-W inhibited thrombin activity, whereas the hot water extract of J-B, J-S, and J-H inhibited blood coagulation factors. In an antiplatelet aggregation activity assay, only the J-W takju lees showed significant inhibition activity. Our results suggest that lees from traditional wine had high potential as a novel antithrombosis agent.

Two new triterpenoid saponins derived from the leaves of Panax ginseng and their antiinflammatory activity

  • Li, Fu;Cao, Yufeng;Luo, Yanyan;Liu, Tingwu;Yan, Guilong;Chen, Liang;Ji, Lilian;Wang, Lun;Chen, Bin;Yaseen, Aftab;Khan, Ashfaq A.;Zhang, Guolin;Jiang, Yunyao;Liu, Jianxun;Wang, Gongcheng;Wang, Ming-Kui;Hu, Weicheng
    • Journal of Ginseng Research
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    • v.43 no.4
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    • pp.600-605
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    • 2019
  • Background: The leaves and roots of Panax ginseng are rich in ginsenosides. However, the chemical compositions of the leaves and roots of P. ginseng differ, resulting in different medicinal functions. In recent years, the aerial parts of members of the Panax genus have received great attention from natural product chemists as producers of bioactive ginsenosides. The aim of this study was the isolation and structural elucidation of novel, minor ginsenosides in the leaves of P. ginseng and evaluation of their antiinflammatory activity in vitro. Methods: Various chromatographic techniques were applied to obtain pure individual compounds, and their structures were determined by nuclear magnetic resonance and high-resolution mass spectrometry, as well as chemical methods. The antiinflammatory effect of the new compounds was evaluated on lipopolysaccharide-stimulated RAW 264.7 cells. Results and conclusions: Two novel, minor triterpenoid saponins, ginsenoside $LS_1$ (1) and 5,6-didehydroginsenoside $Rg_3$ (2), were isolated from the leaves of P. ginseng. The isolated compounds 1 and 2 were assayed for their inhibitory effect on nitric oxide production in LPS-stimulated RAW 264.7 cells, and Compound 2 showed a significant inhibitory effect with $IC_{50}$ of $37.38{\mu}M$ compared with that of NG-monomethyl-L-arginine ($IC_{50}=90.76{\mu}M$). Moreover, Compound 2 significantly decreased secretion of cytokines such as prostaglandin $E_2$ and tumor necrosis factor-${\alpha}$. In addition, Compound 2 significantly suppressed protein expression of inducible nitric oxide synthase and cyclooxygenase-2. These results suggested that Compound 2 could be used as a valuable candidate for medicinal use or functional food, and the mechanism is warranted for further exploration.

Comparison of the accuracy of intraoral scanner by three-dimensional analysis in single and 3-unit bridge abutment model: In vitro study (단일 수복물과 3본 고정성 수복물 지대치 모델에서 삼차원 분석을 통한 구강 스캐너의 정확도 비교)

  • Huang, Mei-Yang;Son, Keunbada;Lee, Wan-Sun;Lee, Kyu-Bok
    • The Journal of Korean Academy of Prosthodontics
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    • v.57 no.2
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    • pp.102-109
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    • 2019
  • Purpose: The purpose of this study was to evaluate the accuracy of three types of intraoral scanners and the accuracy of the single abutment and bridge abutment model. Materials and methods: In this study, a single abutment, and a bridge abutment with missing first molar was fabricated and set as the reference model. The reference model was scanned with an industrial three-dimensional scanner and set as reference scan data. The reference model was scanned five times using the three intraoral scanners (CS3600, CS3500, and EZIS PO). This was set as the evaluation scan data. In the three-dimensional analysis (Geomagic control X), the divided abutment region was selected and analyzed to verify the scan accuracy of the abutment. Statistical analysis was performed using SPSS software (${\alpha}=.05$). The accuracy of intraoral scanners was compared using the Kruskal-Wallis test and post-test was performed using the Pairwise test. The accuracy difference between the single abutment model and the bridge abutment model was analyzed by the Mann-Whitney U test. Results: The accuracy according to the intraoral scanner was significantly different (P < .05). The trueness of the single abutment model and the bridge abutment model showed a statistically significant difference and showed better trueness in the single abutment (P < .05). There was no significant difference in the precision (P = .616). Conclusion: As a result of comparing the accuracy of single and bridge abutments, the error of abutment scan increased with increasing scan area, and the accuracy of bridge abutment model was clinically acceptable in three types of intraoral scanners.

In Vitro Evaluation of Shear Bond Strengths of Zirconia Cerami with Various Types of Cement after Thermocycling on Bovine Dentin Surface (지르코니아 표면 처리와 시멘트 종류에 따른 치면과의 전단 결합 강도 비교 연구)

  • Cho, Soo-Hyun;Cho, In-Ho;Lee, Jong-Hyuk;Nam, Ki-Young;Kim, Jong-Bae;Hwang, Sang-Hee
    • Journal of Dental Rehabilitation and Applied Science
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    • v.23 no.3
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    • pp.249-257
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    • 2007
  • State of problem : The use of zirconium oxide all-ceramic material provides several advantages, including a high flexural strength(>1000MPa) and desirable optical properties, such as shading adaptation to the basic shades and a reduction in the layer thickness. Along with the strength of the materials, the cementation technique is also important to the clinical success of a restoration. Nevertheless, little information is available on the effect of different surface treatments on the bonding of zirconium high-crystalline ceramics and resin luting agents. Purpose : The aim of this study was to test the effects of surface treatments of zirconium on shear bond strengths between bovine teeth and a zirconia ceramic and evaluate differences among cements Material and methods : 54 sound bovine teeth extracted within a 1 months, were used. They were frozen in distilled water. These were rinsed by tap water to confirm that no granulation tissues have left. These were kept refrigerated at $4^{\circ}C$ until tested. Each tooth was placed horizontally at a plastic cylinder (diameter 20mm), and embedded in epoxy resin. Teeth were sectioned with diamond burs to expose dentin and grinded with #600 silicon carbide paper. To make sure there was no enamel left, each was observed under an optical microscope. 54 prefabricated zirconium oxide ceramic copings(Lava, 3M ESPE, USA) were assigned into 3 groups ; control, airborne-abraded with $110{\mu}m$ $Al_2O_3$ and scratched with diamond burs at 4 directions. They were cemented with a seating force of 10 ㎏ per tooth, using resin luting cement(Panavia $F^{(R)}$), resin cement(Superbond $C&B^{(R)}$), and resin modified GI cement(Rely X $Luting^{(R)}$). Those were thermocycled at $5^{\circ}C$ and $55^{\circ}C$ for 5000 cycles with a 30 second dwell time, and then shear bond strength was determined in a universal test machine(Model 4200, Instron Co., Canton, USA). The crosshead speed was 1 mm/min. The result was analyzed with one-way analysis of variance(ANOVA) and the Tukey test at a significance level of P<0.05. Results : Superbond $C&B^{(R)}$ at scratching with diamond burs showed the highest shear bond strength than others (p<.05). For Panavia $F^{(R)}$, groups of scratching and sandblasting showed significantly higher shear bond strength than control group(p<.05). For Rely X $Luting^{(R)}$, only between scratching & control group, significantly different shear bond strength was observed(p<.05). Conclusion : Within the limitation of this study, Superbond $C&B^{(R)}$ showed clinically acceptable shear bond between bovine teeth & zirconia ceramics regardless of surface treatments. For the surface treatment, scratching increased shear bond strength. Increase of shear bond strength by sandblasting with $110{\mu}m$ $Al_2O_3$ was not statistically different.

Anti-allergic activities of Castanea crenata inner shell extracts fermented by Lactobacillus bifermentans (유산균 발효에 의한 율피(Castanea crenata inner shell) 열수추출물의 아토피 피부 질환에 관한 효과 연구)

  • Choi, Mi-Ok;Kim, Bae-Jin;Jo, Seung-Kyeung;Jung, Hee-Kyoung;Lee, Jin-Tae;Kim, Hak-Yoon;Kweon, Dae-Jun
    • Food Science and Preservation
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    • v.20 no.4
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    • pp.583-591
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    • 2013
  • Atopic dermatitis (AD) is a common chronic inflammatory disease associated with a cutaneous hypersensitivity reaction to an allergen. Although the incidence of AD is increasing these days, therapeutics has yet to be developed for its treatment. The aim of this study was conducted in order to compare and investigate the characteristic between the Castanea crenata inner shell extract (CS) and the Castanea crenata inner shell extract fermented by Lactobacillus bifermentans (FCS) for an anti-atopic medication. The total polyphenol and flavonoid contents were similar to CS and FCS. In the DPPH and superoxide anion radical scavenging, the CS and FCS had the potential for antioxidant activities. Both of them did not exhibit cytotoxicity to HS68 cells. The evaluation of the anti-inflammatory activity in Raw264.7 cells demonstrated that the FCS has inhibited the LPS-induced production of nitric oxide as compared to the CS. The anti-atopic dermatitis test was done through the induction of DNCB in AD hairless mice. The FCS has inhibited the development of the atopic dermatitis-like skin lesion by transdermal water loss, melanin and erythema of the skin as compared to the CS. Moreover, the pro-inflammatory cytokine IL-$1{\beta}$ and TNF-${\alpha}$ production in hairless mice were inhibited by the FCS treatment. It indicates that the fermentation of the Castanea crenata inner shell has the potential for the treatment of atopic dermatitis.