• Asian Pacific Journal of Cancer Prevention
• /
• 제16권7호
• /
• pp.2883-2887
• /
• 2015

Background: The calcium-binding S100A4 protein is involved in epithelial to mesenchymal transition, oncogenic transformation, angiogenesis, cytoskeletal integrity, mobility and metastasis of cancer cells. This study aimed to clarify the roles of S100A4 in genesis and progression of glioma. Materials and Methods: S100A4 expression was examined by real-time RT-CPR and Western blot in glioma and paired normal brain tissue (n=69), and compared with clinicopathological parameters of tumors. In addition, glioma U251 cells transfected with an S100A4-expressing plasmid were examined for proliferation by MTT, apoptosis by Annexin V-FITC, and migration and invasion with Transwell chambers. Results: Increased S100A4 mRNA expression was found in gliomas, compared with paired non-tumor tissue (p<0.001). Gradual elevation of overexpression of S100A4 was observed with increasing glioma grade (p<0.001). Astrocytoma showed lower S100A4 mRNA expression than oligodendrogliomas, with glioblastomas having highest values (p<0.001). Similar results were obtained for S100A4 protein, a positive link being found between mRNA and protein expression in gliomas (p<0.001). There was higher growth, lower apoptosis, stronger migration and invasion of S100A4 transfectants than control and mock transfected cells (p<0.001). Conclusions: These findings indicate that up-regulated S100A4 expression is positively linked to pathogenesis, progression and histogenesis of glioma by modulating proliferation, apoptosis, migration and invasion.

• Asian Pacific Journal of Cancer Prevention
• /
• 제13권5호
• /
• pp.2015-2020
• /
• 2012

Oleanolic acid (OA) is a naturally occurring triterpenoid in food materials and is a component of the leaves and roots of Olea europaea, Viscum album L., Aralia chinensis L. and more than 120 other plant species. There are several reports validating its antitumor activity against different cancer cells apart from its hepatoprotective activity. However, antitumor activity against skin cancer has not beed studied well thus far. Hence the present study of effects of OA against HaCaT (immortalized keratinocyte) cells - a cell-based epithelial model system for toxicity/ethnopharmacology-based studies - was conducted. Radical scavenging activity ($DPPH{\cdot}$) and FRAP were determined spectrophotometrically. Proliferation was assessed by XTT assay at 24, 48 and 72 hrs with exposure to various concentrations (12.5-200 ${\mu}M$) of OA. Apoptotic induction potential of OA was demonstrated using a cellular DNA fragmentation ELISA method. Morphological studies were also carried out to elucidate its antitumor potential. The results revealed that OA induces apoptosis by altering cellular morphology as well as DNA integrity in HaCaT cells in a dose-dependent manner, with comparatively low cytotoxicity. The moderate toxicity observed in HaCaT cells, with induction of apoptosis, possibly suggests greater involvement of programmed-cell death-mediated mechanisms. We conclude that OA has relatively low toxicity and has the potential to induce apoptosis in HaCaT cells and hence provides a substantial and sound scientific basis for further validation studies.

• Journal of Microbiology and Biotechnology
• /
• 제29권6호
• /
• pp.863-876
• /
• 2019

Farm animals such as piglets are often affected by environmental stress, which can disturb the gut ecosystem. Antibiotics were commonly used to prevent diarrhea in weaned piglets, but this was banned by the European Union due to the development of antibiotic resistance. However, the use of probiotics instead of antibiotics may reduce the risk posed by pathogenic microorganisms and reduce the incidence of gastrointestinal diseases. Therefore, this study was conducted to investigate the effects of Lactobacillus casei Zhang on the mechanical barrier and immune function of early-weaned piglets infected using Escherichia coli K88 based on histomorphology and immunology. Fourteen-day-old weaned piglets were divided into a control group and experimental groups that were fed L. casei Zhang and infected with E. coli K88 with or without prefeeding and/or postfeeding of L. casei Zhang. The L. casei Zhang dose used was $10^7CFU/g$ diet. Jejunum segments were obtained before histological, immunohistochemical, and western blot analyses were performed. In addition, the relative mRNA expression of toll receptors and cytokines was measured. Piglets fed L. casei Zhang showed significantly increased jejunum villus height, villus height-crypt depth ratio, muscle thickness, and expression of proliferating cell nuclear antigen and tight junction proteins ZO-1 and occludin. The use of L. casei Zhang effectively reduced intestinal inflammation after infection. We found that L. casei Zhang feeding prevented the jejunum damage induced by E. coli K88, suggesting that it may be a potential alternative to antibiotics for preventing diarrhea in early-weaned piglets.

• 한국식품영양학회지
• /
• 제35권6호
• /
• pp.513-520
• /
• 2022

The purpose of this study was to investigate the effect of Cheonggukjang pills with aronia and blueberries on dextran sulfate sodium (DSS)-induced colitis in mice. There have been several reports that Cheonggukjang is effective for intestinal health, but the efficacy of Cheonggukjang containing fruits has not yet been reported. In this study, we showed the effect of cheonggukjang pills with blueberries and aronia (CPBA) on DSS-induced colitis in BALB/c mice. CPBA was obtained from Soonchang Moonokae foods and orally administered once a day for 2 weeks before DSS treatment. Colitis was induced in mice by feeding 5% (w/v) DSS drinking water for 7 days. The results showed that CPBA treatment significantly alleviated DSS-induced disease activity index associated with a decrease in colon length. CPBA improved DSS-induced histological changes and intestinal epithelial barrier integrity in mice colon. In addition, CPBA administration significantly reduced the levels of DSS-mediated interferon-γ and interleukin-6 in serum and tumor necrosis factor-α in colon tissue. Moreover, the gene expression of COX-2 and iNOS, which are factors involved in inflammatory signaling, was significantly reduced by CPBA treatment. These results suggest that CPBA have a protective effect against DSS-induced mice colitis and may be a candidate for colitis treatment.

• Animal Bioscience
• /
• 제36권9호
• /
• pp.1403-1413
• /
• 2023

Objective: Intestinal alkaline phosphatase (IAP) maintains intestinal homeostasis by detoxifying bacterial endotoxins and regulating gut microbiota, and lipid absorption. Antibiotics administered to animals can cause gut dysbiosis and barrier disruption affecting animal health. Therefore, the present study sought to investigate the role of IAP in the intestinal environment in dysbiosis. Methods: Young male mice aged 9 weeks were administered a high dose of antibiotics to induce dysbiosis. They were then sacrificed after 4 weeks to collect the serum and intestinal organs. The IAP activity in the ileum and the level of cytokines in the serum samples were measured. Quantitative real-time polymerase chain reaction analysis of RNA from the intestinal samples was performed using primers for tight junction proteins (TJPs) and proinflammatory cytokines. The relative intensity of IAP and toll-like receptor 4 (TLR4) in intestinal samples was evaluated by western blotting. Results: The IAP activity was significantly lower in the ileum samples of the dysbiosis-induced group compared to the control. The interleukin-1 beta, interleukin-6, and tumor necrosis factor-alpha concentrations were significantly higher in the ileum samples of the dysbiosis-induced group. The RNA expression levels of TJP2, claudin-3, and claudin-11 showed significantly lower values in the intestinal samples from the dysbiosis-induced mice. Results from western blotting revealed that the intensity of IAP expression was significantly lower in the ileum samples of the dysbiosis-induced group, while the intensity of TLR4 expression was significantly higher compared to that of the control group without dysbiosis. Conclusion: The IAP activity and relative mRNA expression of the TJPs decreased, while the levels of proinflammatory cytokines increased, which can affect intestinal integrity and the function of the intestinal epithelial cells. This suggests that IAP is involved in mediating the intestinal environment in dysbiosis induced by antibiotics and is an enzyme that can potentially be used to maintain the intestinal environment in animal health care.

• Tuberculosis and Respiratory Diseases
• /
• 제40권3호
• /
• pp.236-249
• /
• 1993

연구배경 : 정상 폐상피세포에서는 항상 생성되고 있는 활성산소(oxygen radical)에 의한 유해작용에 노출되어 있고, 이들 유해 산소들은 폐기종과 같은 폐질환의 원인 기전으로 생각되고 있다. 본 연구에서는 이 방법을 이용하여 만든 폐상피세포 단일막에서 전기생리학적인 관점에서 물질의 이동지표인 short-circuit current(Isc)와 조직저항(R)에 대한 활성산소의 하나인 $H_2O_2$(hydrogen peroxide)가 어떤 영향을 미치는지를 연구함으로서 세포생리학적 기전을 구명하고자 한다. 방법 : Tissue culture-treated polycarbonant membrane filter 에서 배양시킨 쥐 제 2 형 폐상피세포 배양 단일막에서 $H_2O_2$의 능동적 이온 이동 (Isc) 과 수동적 용질이동에 대한 조직저항(R)에 미치는 효과를 관찰하였다. 배양 제 3 일과 제 4 일째 단일막을 수정된 Ussing chamber에 설치하고 막 양측에 HEPES-buffered Ringer 용액으로 incubation 하였다. 외부에서 0~100 mM 농도의 $H_2O_2$를 apical 또는 basolateral side에 작용시켜 Isc와 R의 변화를 관찰하였다. 폐상피세포 장벽이 외부의 $H_2O_2$에 대하여 방어작용을 가지는 세포내 catalase 활성도를 측정하고, catalase 억제제인 aminotriazol(ATAZ) 20 mM의 효과도 함께 관찰하였다. 결과 : 이 단일막은 형태학적으로 보아서 in vivo 에서의 포유류 제 1 형 폐상피세포 장벽의 특성을 나타내고 세포들 사이는 tight junction을 이루며(조직저항 R: 2,000 ohm-$cm^2$ 이상) sodium ion의 능동적 이동 (Isc: 5 ${\mu}A/cm^2$)을 보였다. $H_2O_2$는 dose-dependent 양식으로 Isc와 R 모두 감소시켰다. Apical side에 작용하는 $H_2O_2$에 있어서는 60분에 50% 억제하는 농도인 $ED_{50}$는 Isc와 R은 약 4mM 이었으나 basolateral side의 경우는 약 0.04mM 로서 그 작용 강도는 apical에 비하여 약 100배 정도 더 컸다. ATAZ 존재시 apical side의 $ED_{50}$는 0.4mM로 감소하였으나 basolateral side의 경우 변화가 없었다. $H_2O_2$의 제거율은 apical 또는 basolateral side 어느 쪽에 존재하든 같았으며, 세포내 catalase 활성도는세포배양기간이 길어짐에 따라 증가함을 보였다. 결론 : 이상의 실험결과는 basolateral side에 작용하는 $H_2O_2$는 세포내 막구성성분 중 basolateral 측에 존재하는 곳에(예, $Na^+,\;K^+$-APTase) 직접 장애를 미칠 것으로 생각된다. 한편 apical side에 작용하는 $H_2O_2$는 막성분에 도달하기 전에 세포내에 존재하는 catalase에 의하여 대부분 그 작용을 잃게 된다. 결론적으로 Isc와 R로 측정된 폐상피세포 장벽의 특성은 $H_2O_2$에 의하여 손상을 받고 apical side 보다는 basolateral side 측정이 더 손상을 잘 받게 된다.

• 생명과학회지
• /
• 제27권4호
• /
• pp.482-499
• /
• 2017

Allergy 질환은 지난 십여년 동안 개발도상국을 포함해서 전 세계적으로 증가하고 있다. Allergy 염증 반응은 수지상 세포와 같은 항원제시 세포에 의한 allergy 항원섭취를 시작으로 하여 Th2 면역 반응에 의해서 일어난다. 장내 미생물은 신체의 대사나 생리적 기능을 조절하고, 생애 초기의 면역 체계 성숙과 일생 동안 면역 체계 항상성 및 상피세포 총체성에 기여한다. Bifidobacteria는 Th1/Th2 balance에 strain-specific 한 면역 자극 특성을 가지며, TSLP와 IgE 발현을 억제 시키고 Flg과 FoxP3 발현을 촉진 시켜 allergy를 완화시킨다. 또한 Unmethylated CpG motif ODN은 B 세포와 수지상 세포의 TLR9에 의해 인식 되어 선천성과 적응성 면역 반응을 유도하고, Clostridium butyricum에 의해서 생산된 butyrate는 수지상 세포의 anti-inflammatory 유전자의 발현을 유도하기 위해 GPR109a signaling pathway를 활성화시키고, GPR43 활성화를 통하여 tTreg 세포 proliferation을 직접 자극하거나 HADC 활성을 억제시켜 Foxp3 gene intronic enhancer의 histone H3 acetylation을 통해 naive $CD4^+$ T 세포를 pTreg 세포로 분화시킨다.

• 폴리머
• /
• 제35권4호
• /
• pp.289-295
• /
• 2011

망막조직공학을 위한 생체 재료는 기계적 안정성, 생체적합성, 낮은 분해속도 등을 포함하여 in vivo에서 잠재적인 유용성을 위한 몇 가지 중요한 특징이 입증되어야 한다. 실크 필름 생체재료는 이러한 기능적인 요구에 맞게 디자인되었다. 0, 10, 20, 40, 및 80 wt%의 실크가 함유된 천연합성물질과 하이브리드화된 silk/PLGA 필름을 용매 증발법으로 제조하였다. 1, 2, 및 3일 후에 부착된 세포 수를 확인하기 위해 MTT 분석을 하였고 SEM을 통해 필름에 부착된 세포 모폴로지를 확인하였다. 또한, mRNA 발현정도를 알아보기 위해 retinal pigment epithelitun(RPE) 세포의 프라이머인 RPE65를 사용하여 RT-PCR을 실시하였고 RPE 세포의 특정 단백질인 cytokeratin의 발현을 확인하고 세포의 증식을 비교하기 위해 면역화학염색을 실시하였다. 본 실험을 통해 실크/PLGA 필름에서 20~40 wt% 실크를 함유한 경우에 RPE 세포의 부착과 증식에 가장 좋은 영향을 미치는 것을 확인하였다.

• Parasites, Hosts and Diseases
• /
• 제28권1호
• /
• pp.45-52
• /
• 1990

인체 기생 장흡충류의 하나인 호르텐스극구흠충(Echinostoma hortense)의 병원성을 알아 보기 위하여 흰쥐에 퍼낭유충을 실험 감염시킨 후 경시적으로 숙주의 장병변을 관찰하였다. 피낭유충은 실험적으로 감염시킨 올챙이에서 분리한 것을 사용하였고 Sprague-Dawley계 흰쥐 18마리에 각각 200개씩 감염시킨 후 1일, 3일, 7일, 11일, 22일 및 44일에 3마리씩 희생시키고 위 유문으로부터 1cm, 3cm, 5cm, 8cm 및 30cm 부위에서 각각 장 절편을 채취하여 병리 조직학적 관찰을 시 행하였다. 관찰 결과 충체는 감염 1∼3일에는 융모 사이 (infervillous space)에서, 감염 7일부터 44일까지는 주로 내강(lumen)에서 발견되었고 구흡반 및 복흡반으로 융모를 흡입(sucking, 파괴하고 있는 것이 관찰되었다. 장 조직의 병변은 융모 위축(villous atrophy)과 선와 증식 (crypt hyperplasia)이 특징적 소견이었다. 융모의 변화는 끝(tip)의 둔화, 유착, 소실 등이었고 융모1선와 높이의 비율 이 3 : 1에서 1 : 1까지 감소되었다. 기질의 변화로는 염증 세포의 침윤, 충혈, 부종, 섬유화 등이 관 찰되었다. 배세포는 감염 11일 경부터 점차 증가하는 소견을 보였다. 전체적으로 장 병변은 소장 상 부에서 카장 심하였고 감염 1∼3일부터 관찰되기 시작하였으며 감염기간의 경과에 따라 점차 심화 되었다. 이상의 결과를 종합할 때 호르텐스극구흡충에 감염된 횐쥐의 장 병변은 점막층에 국한되어 있으나 융모의 위축 및 파괴가 매우 심하며 감염 44일까지도 병변이 지속되고 있음을 알 수 있었다. Key worths: Echinostoma hortense, rat intestinal pathology, villous atrophy, crypt hyperplasia로 출아묘는 이앙기간이 단축되므로 중묘보다는 다소 늦게 파종하고 이앙기는 중묘 적기에 하는 냉해의 우려도 없어 좋을 것으로 생각된다. 6. 간장은 중묘보다 어린모 이앙재배에서 3∼4cm 기었는데 이는 천식이 되어 초기생육이 왕성하므로 과번무 되었기 때문으로 생각되며 지나친 천식은 태풍 등으로 도복의 위험성이 있다. 7. 수량성은 조식재배의 경우 어린모는 중묘와 비슷한 경향이며 직파재배 보다 다소 증수되었다.질소함량은 경장, 건물중, 근류수 및 근류중들과, 그리고 줄기중의 allantoin 질소함량은 근류수 및 근류중들과 각각 유의적인 정의 상관관계를 나타내었다. 희미해졌으며 담배는 500 $\mu$ E m$^{-2}$ sec$^{-1}$에서 인삼은 1000 $\mu$ E m$^{-2}$ sec$^{-1}$에서 47KD이었다.50일영이상)에서 35.3％ 및 유추(30 일영미만)에서 17.3％의 순이었다. 이중 세균성 및 기생충성 질병은 육성계에서의 검색률이 가장 높았으나, 바이러스성 질병은 육성계 및 성계에서 거의 같은 비율로 높았으며 곰팡이성 질병은 유추에서 가장 높은 검색률을 나타내었다.17) 사료비절감은 생산비의 60∼70％를 차지하고 있으므로 공동구인 자가배합을 하여 자기실정에 맞는 경영체질로 운영하고 있었다. (18) 인건비가 생산비의 15∼20％의 비율이며 또한 사람 구득난이라 가족노동을 최대한 활용, 관리를 하고 있으며 생력화를 위해 기계화, 자동화되어 가고 있었다. 애정과 정성심있는 관리로 닭을 스트레스를 주지 않게 하기 위해 가족관이, 기계자동화가 중요한 역할을 하고 있었다. (19) 시설기구와 육성상각비를 줄이기 위해 관리자의 변동이 없고, 종신직장으로서 긍지와 자부심을 가지고 늑무하고 있었다.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• 제32권3호
• /
• pp.189-199
• /
• 2006

Purpose of study: Partial thickness skin graft is the golden standard regimen for full-thickness skin defect caused by burn or trauma. However, in case of extensive burns of more than 50% of total body surface area, the donor site is not sufficient to cover all defects. As a second choice, allograft, xenograft and synthetic materials have been used to treat skin defect. Among them the amniotic membrane(AM) was used as a biological dressing for centuries because of its potential for wound healing. In this study, quantification of EGF in AM and effect of AM-collagen complex on full thickness skin defects was examined. Materials & Methods: The concentration of EGF in fresh, deep frozen and freeze-dried AM was evaluated by ELISA. EGF-R immunostaining was performed in freeze-dried AM. SD rats weighing 250${\sim}$300g was used for wound healing experiment. Three full thickness skin defects(28mm diameter) were made on dorsal surface of SD rat. The control group was covered by Vaselin gauze and AM-collagen complex and $Terudermis^{(R)}$. was grafted in two other defects. Healing area, Cinamon's score were evaluated before biopsy. Grafted sites were retrieved at 3 days, 1 week, 2 weeks and 4 weeks after operation. H & E and Factor VIII immunohistochemical stain was performed to evaluate the microscopic adhesion and structural integrity and microvessel formation. Results: 1. EGF concentration of fresh, deep frozen and freeze-dried AM showed similar level and EGF-R was stained in epithelial layer of freeze-dried AM. 2. At 4 weeks after grafting, the healing area of AM-collagen and Terudermis group was 99.29${\pm}$0.71% and 99.19${\pm}$0.77 of original size. However, that of control group was 24.88${\pm}$2.90. 3. The Cinamon's score of AM-Collagen and $Terudermis^{(R)}$. group at 4 weeks was 15.6${\pm}$1.26 and 14.6${\pm}$3.13 and that of control group was 3.7${\pm}$0.95. Significant difference was observed among control and experimental groups(p<0.05). 4. Histologic examination revealed that AM protected leukocyte infiltration and epithelial migration was nearly completed at 4 weeks. $Terudermis^{(R)}$. group showed mild neutrophil infiltration until 2 weeks and completion of epithelization at 4 weeks. Control group showed massive leukocyte infiltration until 4 weeks. 5. Microvessels were increased sharply at 1 week and control group at 1 and 4 week showed significant differences with $Terudermis^{(R)}$. group of same interval(p<0.05) but no differences were found with AM group(p<0.05). Conclusion: EGF and EGF-R were well preserved in freeze-dried AM. AM attached to collagen acted as excellent biologic dressing which had similar effect with $Terudermis^{(R)}$. AM showed anti-inflammatory action and healing was completed at 4 weeks after full-thickness skin defect.