• 동의생리병리학회지
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• 제19권6호
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• pp.1666-1672
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• 2005

Oriental medicinal drugs have a broad spectrum of clinical use for the cure of nervous system diseases including brain ischemic damages or neuropathies. Yet, specific drugs or drug components used in the oriental medicine in relation to none fiber regeneration are not known. In the present study, possible growth promoting effects of oriental medicinal drugs were investigated in the injured sciatic nerve system in the rat. By immunofluorescence staining, we found that Jahageo (JHG, Hominis placenta) increased Induction levels of axonal growth associated protein GAP-43 in the rat sciatic none. Small growth promoting activity was found in Golsebo (GSB, Drynariae rhizoma) and Baikhasuo (BHSO, Polygoni multiflori radix) drugs. JHG also increased cell cycle protein Cdc2 levels in the injured area of the sciatic nerves. Immunofluorescence staining indicated that induced Cdc2 protein was mostly localized in the Schwann cells in the injury area, implying that JHG activity might be related to increased Schwann cell proliferation during axonal regeneration. Moreover, levels of phospho-extracellular signal-regulated (ERK) pathway in the injured neNes were elevated by JHG treatment while levels of total ERK were unaltered. In vivo measurement of axonal regeneration using retrograde tracer showed that JHG, GSB and BHSO significantly enhanced Dil-labeled regenerating motor neurons compared with saline control. The present data suggest that oriental medicinal drugs such as JHG, GSB, and BHSO may be a useful target for developing specific drugs of axonal regeneration.

• 대한수의학회지
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• 제33권2호
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• pp.249-254
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• 1993

설사로 폐사한 자돈의 장 절편을 이용하여 돼지 유행성 설사바이러스(PEDV), 돼지 전염성 위장염 바이러스(TGEV), 돼지 로타 바이러스(PRV)에 대한 병인학적 검사를 형광항체반응을 통하여 조사하였던 바 돼지 유행성 설사 바이러스의 감염에 의한 자돈의 폐사를 확인하였다. 간접형광항체검사시 돼지 유행성 설사 바이러스에 대한 양성반응을 보인 가검재료를 이용 Vero세포에 연속 계대한 후 plaque assay를 통하여 크로닝된 돼지 유행성 설사 바이러스 KPEDV-9주를 작성하였다. 돼지 유행성 설사 바이러스에 대한 면역혈청과 바이러스가 분리된 농장에서 채취된 돼지 혈청을 이용 돼지 유행성 설사 바이러스에 대한 구조단백성분을 분석하였던 바 88K(M.W.), 74K, 70K, 58~54K, 54~46K, 44~40K 및 33~32K에 상당하는 단백성분을 검출할 수 있었다.

• 혜화의학회지
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• 제29권1호
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• pp.18-25
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• 2020

Objective: The goal of this study is to investigate whether peripheral axonal regeneration is affected by diabetes in experimental animals. Method: Sprague Dawely rat was injected with 45~50 mg/kg of streptozotocin (STZ) to generate an animal model of diabetes. Three months after STZ injection, sciatic nerve (2 cm length) was removed and the same length of nerve segments from STZ-injected animal or from control animal (CTL) was transplanted into STZ-injected animals (STZ-STZ and STZ-CTL respectively). Similarly, sciatic nerve segments from STZ-injected animal or from control animal were grafted into the control animals (CTL-STZ and CTL-CTL respectively). All animals were sacrificed 2 weeks after transplantation. Sciatic nerve sections were prepared and subjected to immunofluorescence staining analysis. Results: Immunofluorescence staining for NF-200 showed that distal elongation of regenerating axons reached 40~80% of proximal neve in both CTL-STZ and CTL-CTL groups. However, distal elongation in both STZ-STZ and STZ-CTL groups were 20~60% of proximal nerve. Furthermore, measurement of axonal regeneration after immuno-staining with SCG10 showed that the scores of distal elongation relative to proximal nerve were 50~90% in CTL-CTL and CTL-STZ groups and 10-60% in STZ-CTL and STZ-STZ. Conclusions: Our data showed that the levels of axonal regeneration were not affected irrespective of whether they were from STZ- or CTL graft, but were greatly reduced when the nerves were transplanted into the STZ host.

• 생명과학회지
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• 제22권10호
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• pp.1415-1419
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• 2012

Thymosin ${\beta}4$ 는 대장암에서 암 줄기세포 마커인 CD133을 지닌 세포에서 지니지 않은 세포에 비해 강하게 발현된다고 보고되어 있다. 본 연구에서는 thymosin ${\beta}4$와 줄기세포 마커인 CD133의 상관관계를 정상 위 조직에서 관찰하였다. Thymosin ${\beta}4$와 CD133의 발현 양상은 tissue microarray 조직상에서 면역화학적 방법으로 관찰하였으며 thymosin ${\beta}4$와 CD133의 존재 위치는 면역형광 염색법 및 confocal microscope를 사용하여 조사하였다. Thymosin ${\beta}4$와 CD133은 동일한 양상으로 발현되었으며 모두 위의 선조직에서 강하게 발현되었다. 면역 형광 염색법으로 두가지 단백질을 동시에 염색한 결과 두 단백질이 동일한 위치에서 함께 존재하는 것으로 규명되었다. 이러한 결과는 thymosin ${\beta}4$와 CD133은 정상위의 선조직에서 발현되며 두 단백질의 발현 양상 및 위치가 동일하여 서로 긴밀한 상호작용을 할 가능성을 제시하고 있다.

• 한국발생생물학회지:발생과생식
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• 제24권3호
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• pp.177-185
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• 2020

Although many aquaporin (AQP) transcripts have been demonstrated to express in the female reproductive tract, the defined localizations and functions of AQP subtype proteins remain unclear. In this study, we investigated the expression of AQP1, AQP3, AQP5, AQP6, and AQP9 proteins in female reproductive tract of mouse and characterized their precise localizations at the cellular and subcellular levels. Immunofluorescence analyses for AQP1, AQP3, AQP6, and AQP9 showed that these proteins were abundantly expressed in female reproductive tract and that intense immunoreactivities were observed in mucosa epithelial cells with a subtype-specific pattern. The most abundant aquaporin in both vagina and uterine cervix was AQP3. Each of AQP1, AQP3, AQP6, and AQP9 exhibited its distinct distribution in stratified squamous or columnar epithelial cells. AQP9 expression was predominant in oviduct and ovary. AQP1, AQP3, AQP6, and AQP9 proteins were mostly seen in apical membrane of ciliated epithelial cells of the oviduct as well as in both granulosa and theca cells of ovarian follicles. Most of AQP subtypes were also expressed in surface epithelial cells and glandular cells of endometrium in the uterus, but their expression levels were relatively lower than those observed in the vagina, uterine cervix, oviduct and ovary. This is the first study to investigate the expression and localization of 5 AQP subtype proteins simultaneously in female reproductive tract of mouse. Our results suggest that AQP subtypes work together to transport water and glycerol efficiently across the mucosa epithelia for lubrication, proliferation, energy metabolism and pH regulation in female reproductive tract.

• Journal of Korean Neurosurgical Society
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• 제55권3호
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• pp.131-135
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• 2014

Objective : With the growing interests of bacteria as a targeting vector for cancer treatment, diverse genetically engineered Salmonella has been reported to be capable of targeting primary or metastatic tumor regions after intravenous injection into mouse tumor models. The purpose of this study was to investigate the capability of the genetically engineered Salmonella typhimurium (S. typhimurium) to access the glioma xenograft, which was monitored in mouse brain tumor models using optical bioluminescence imaging technique. Methods : U87 malignant glioma cells (U87-MG) stably transfected with firefly luciferase (Fluc) were implanted into BALB/cAnN nude mice by stereotactic injection into the striatum. After tumor formation, attenuated S. typhimurium expressing bacterial luciferase (Lux) was injected into the tail vein. Bioluminescence signals from transfected cells or bacteria were monitored using a cooled charge-coupled device camera to identify the tumor location or to trace the bacterial migration. Immunofluorescence staining was also performed in frozen sections of mouse glioma xenograft. Results : The injected S. typhimurium exclusively localized in the glioma xenograft region of U87-MG-bearing mouse. Immunofluorescence staining also demonstrated the accumulation of S. typhimurium in the brain tumors. Conclusion : The present study demonstrated that S. typhimurium can target glioma xenograft, and may provide a potentially therapeutic probe for glioma.

• Clinical and Experimental Pediatrics
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• 제64권8호
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• pp.422-429
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• 2021

Background: Carnosine has antioxidative and neuroprotective properties against hypoxic-ischemic (HI) brain injury. Hypothermia is used as a therapeutic tool for HI encephalopathy in newborn infants with perinatal asphyxia. However, the combined effects of these therapies are unknown. Purpose: Here we investigated the effects of combined carnosine and hypothermia therapy on HI brain injury in neonatal rats. Methods: Postnatal day 7 (P7) rats were subjected to HI brain injury and randomly assigned to 4 groups: vehicle; carnosine alone; vehicle and hypothermia; and carnosine and hypothermia. Carnosine (250 mg/kg) was intraperitoneally administered at 3 points: immediately following HI injury, 24 hours later, and 48 hours later. Hypothermia was performed by placing the rats in a chamber maintained at 27℃ for 3 hours to induce whole-body cooling. Sham-treated rats were also included as a normal control. The rats were euthanized for experiments at P10, P14, and P35. Histological and morphological analyses, in situ zymography, terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assays, and immunofluorescence studies were conducted to investigate the neuroprotective effects of the various interventional treatments. Results: Vehicle-treated P10 rats with HI injury showed an increased infarct volume compared to sham-treated rats during the triphenyltetrazolium chloride staining study. Hematoxylin and eosin staining revealed that vehicle-treated P35 rats with HI injury had decreased brain volume in the affected hemisphere. Compared to the vehicle group, carnosine and hypothermia alone did not result in any protective effects against HI brain injury. However, a combination of carnosine and hypothermia effectively reduced the extent of brain damage. The results of in situ zymography, TUNEL assays, and immunofluorescence studies showed that neuroprotective effects were achieved with combination therapy only. Conclusion: Carnosine and hypothermia may have synergistic neuroprotective effects against brain damage following HI injury.

• Journal of Periodontal and Implant Science
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• 제49권3호
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• pp.193-204
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• 2019

Purpose: The reaction of cells to a titanium implant depends on the surface characteristics of the implant which are affected by decontamination. The aim of this study was to evaluate the cytocompatibility of titanium disks treated with various decontamination methods, using salivary bacterial contamination with dental pellicle formation as an in vitro model. Methods: Sand-blasted and acid-etched (SA) titanium disks were used. Three control groups (pristine SA disks [SA group]; salivary pellicle-coated SA disks [pellicle group]; and biofilm-coated, untreated SA disks [NT group]) were not subjected to any decontamination treatments. Decontamination of the biofilm-coated disks was performed by 14 methods, including ultrasonic instruments, rotating instruments, an air-powder abrasive system, a laser, and chemical agents. MG63 cells were cultured in the presence of the treated disks. Cell proliferation assays were performed on days 2 and 5 of cell culture, and cell morphology was analyzed by immunofluorescence and scanning electron microscopy (SEM). A vascular endothelial growth factor (VEGF) assay was performed on day 5 of culture. Results: The cell proliferation assay revealed that all decontaminated disks, except for the 2 groups treated using a plastic tip, showed significantly less cell proliferation than the SA group. The immunofluorescence and SEM analyses revealed that most groups showed comparable cell density, with the exception of the NT group, in which the cell density was lower and bacterial residue was observed. Furthermore, the cells grown with tetracycline-treated titanium disks showed significantly lower VEGF production than those in the SA group. Conclusions: None of the decontamination methods resulted in cytocompatibility similar to that of pristine SA titanium. However, many methods caused improvement in the biocompatibility of the titanium disks in comparison with the biofilm-coated, untreated titanium disks. This suggests that decontamination is indispensable for the treatment of peri-implantitis, even if the original biocompatibility cannot be restored.

• Journal of Microbiology and Biotechnology
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• 제29권8호
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• pp.1281-1287
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• 2019

Nattokinase (NK, E.C. 3.4.21.62) is a serine protease produced by Bacillus subtilis natto that shows promise for the treatment of thrombotic disease. In this study, we assessed the effects of NK on the development of hepatocellular carcinoma (HCC), a principal malignancy of the liver that causes morbidity and mortality worldwide. Crude extracts of NK (NCE) were isolated from fermentation medium by centrifugation and separated into three fractions (<10 K, 100~30 K and >30K). Orthotopic HCC mouse models were established and NCE was administered by oral gavage. H&E staining was performed to examine the pathology of HCC livers. Immunohistochemistry and immunofluorescence were used to evaluate FOXM1, CD31, CD44 and vimentin expression in the liver. Compared to PBS groups, NCE increased the survival rates of HCC-bearing mice to 31% and decreased ascites. Low-intensity ultrasound imaging showed that the hypoechoic mass area was lower in NCE-treated mice and that tumor growth significantly decreased. IHC staining showed that the expression of FOXM1 was inhibited by NCE treatment. Immunofluorescence results revealed lower levels of CD31, CD44 and vimentin in the NCE groups. Taken together, these data demonstrate that NCE from Bacillus subtilis natto improves survival and inhibits tumor growth in HCC mice.

• 대한본초학회지
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• 제36권5호
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• pp.59-67
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• 2021

Objectives : Hibiscus (Hibiscus sabdariffa L.) is rich in antioxidants such as flavonoids and anthocyanins and is known to have anti-inflammatory activity and anti-aging function of the skin, but there is no study on its effect on the skin barrier. This study aim to investigate the positive effect on the skin barrier by confirming the effect of water extracts of Hibiscus sabdariffa L. (WEHS) on the tight junction-related gene expression. Methods : The antioxidant efficacy of WEHS was investigated through ABTS and DPPH assays, and 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium was performed to examine the effect on cell viability. quantitative Reverse transcription polymerase chain reaction and immunoblot analysis were performed to confirm the effect of WEHS on the expression of tight junction-related genes, and immunofluorescence microscopy was used to confirm the movement of Claudin 1 protein into tight junctions. Results : WEHS showed strong antioxidant activity and induced an increase in both mRNA and protein expression levels of Claudin 1 among tight junction-related genes. The strong localization of Claudine 1 protein increased by WEHS to the tight junction was confirmed by immunofluorescence microscopy. Conclusions : Hibiscus was confirmed through this study to show antioxidant activity and the function of promoting the expression of the tight junction Claudin 1 gene, suggesting that Hibiscus can be used as a material for the prevention and treatment of skin moisturizing and atopy, which have an important influence on tight junction.