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Assessment of neural activity in the specific brain area is critical for understanding the circuit mechanisms underlying altered brain function and behaviors. A number of immediate early genes (IEGs) that are rapidly transcribed in neuronal cells in response to synaptic activity have been used as markers for neuronal activity. However, protein detection of IEGs requires translation, and the amount of newly synthesized gene product is usually insufficient to detect using western blotting, limiting their utility in western blot analysis of brain tissues for comparison of basal activity between control and genetically modified animals. Here, we show that the phosphorylation status of eukaryotic elongation factor-2 (eEF2) rapidly changes in response to synaptic and neural activities. Intraperitoneal injections of the GABA A receptor (GABAAR) antagonist picrotoxin and the glycine receptor antagonist brucine rapidly dephosphorylated eEF2. Conversely, potentiation of GABAARs or inhibition of AMPA receptors (AMPARs) induced rapid phosphorylation of eEF2 in both the hippocampus and forebrain of mice. Chemogenetic suppression of hippocampal principal neuron activity promoted eEF2 phosphorylation. Novel context exploration and acute restraint stress rapidly modified the phosphorylation status of hippocampal eEF2. Furthermore, the hippocampal eEF2 phosphorylation levels under basal conditions were reduced in mice exhibiting epilepsy and abnormally enhanced excitability in CA3 pyramidal neurons. Collectively, the results indicated that eEF2 phosphorylation status is sensitive to neural activity and the ratio of phosphorylated eEF2 to total eEF2 could be a molecular signature for estimating neural activity in a specific brain area. Supplementary Information The online version contains supplementary material available at 10.1186/s13041-021-00852-0.

OBJECTIVE Discovery of a novel antibody would enable diagnosis and early treatment of autoimmune encephalitis. The aim was to discover a novel antibody targeting a synaptic receptor and characterize the pathogenic mechanism. METHOD We screened for unknown antibodies in serum and cerebrospinal fluid samples from autoimmune encephalitis patients. Samples with reactivity to rat brain sections and no reactivity to conventional antibody tests underwent further processing for antibody discovery, using immunoprecipitation to primary neuronal cells, mass-spectrometry analysis, an antigen-binding assay on an antigen-overexpressing cell line, and an electrophysiological assay with cultured hippocampal neurons. RESULTS Two patients had a novel antibody against CaV α2δ (voltage-gated calcium channel alpha-2/delta subunit). The patient samples stained neuropils of the hippocampus, basal ganglia, and cortex in rat brain sections and bound to a CaV α2δ-overexpressing cell line. Knockdown of CaV α2δ expression in cultured neurons turned off the immunoreactivity of the antibody from the patients to the neurons. The patients were associated with preceding meningitis or neuroendocrine carcinoma and responded to immunotherapy. In cultured neurons, the antibody reduced neurotransmitter release from presynaptic nerve terminals by interfering with tight coupling of calcium channels and exocytosis. INTERPRETATION Here, we discovered a novel autoimmune encephalitis associated with anti-CaV α2δ antibody. Further analysis of the antibody in autoimmune encephalitis might promote early diagnosis and treatment. ANN NEUROL 2021;89:740-752.

Simple Summary We here report the first detection of Enterocytozoon hepatopenaei (EHP) in cultured Pacific whiteleg shrimp (Litopenaeus vannamei) with a growth disorder in Korea. The histopathology, electron microscopy and DNA phylogeny indicated that the EHP is a unique strain first recorded in Korea. This finding highlights the need for closer monitoring and surveillance to control EHP in aquaculture to prevent disastrous economic losses. Abstract The consumption of cultured crustaceans has been steadily increasing, and Pacific whiteleg shrimp (Litopenaeus vannamei) are major cultivated invertebrates worldwide. However, shrimp productivity faces a variety of challenges, mainly related to outbreaks of lethal or growth retardation-related diseases. In particular, hepatopancreatic microsporidiosis caused by the microsporidian parasite Enterocytozoon hepatopenaei (EHP) is an important disease associated with growth retardation in shrimp. Here, we report the detection of EHP through histopathological, molecular and electron microscopy methods in the hepatopancreas of Pacific whiteleg shrimp with growth disorder in a South Korean farm. Phylogenetic analysis showed a clade distinct from the previously reported EHP strains isolated in Thailand, India, China and Vietnam. An EHP infection was not associated with inflammatory responses such as hemocyte infiltration. Although EHP infection has been reported worldwide, this is the first report in the shrimp aquaculture in Korea. Therefore, an EHP infection should be managed and monitored regularly for effective disease control and prevention.

Oxidative stress is a major contributor to muscle aging and loss of muscle tissue. Jakyakgamcho-tang has been used in traditional Eastern medicine to treat muscle pain. Here, we compared various solvent-based Jakyakgamcho-tang extracts in terms of their effects against hydrogen peroxide-induced oxidative stress in murine C2C12 skeletal muscle cells. Total phenolic content and total flavonoid content in 30% ethanol extracts of Jakyakgamcho-tang were higher than those of water extracts of Jakyakgamcho-tang. Ethanol extracts of Jakyakgamcho-tang had stronger antioxidant and 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid and 2,2´-diphenyl-1-picrylhydrazyl-scavenging activity than water extracts of Jakyakgamcho-tang. The ethanol extract of Jakyakgamcho-tang inhibited peroxide-induced cell viability and intracellular reactive oxygen species generation more effectively than the water extract of Jakyakgamcho-tang in a dose-dependent manner. These results suggest that the ethanol extract of Jakyakgamcho-tang is relatively more efficacious at protecting against oxidative stress-induced muscle cell death because it prevents reactive oxygen species generation in C2C12 cells. Moreover, the current study indicated that the effective dose of the ethanol extract of Jakyakgamcho-tang required to alleviate muscle pain might be lower than that required for Jakyakgamcho-tang.

Currently, injuries to customers due to health functional foods are annually increasing. To evaluate the antigenicity of Korean red ginseng mixture (KRGM), we tested for systemic anaphylactic shock and passive cutaneous anaphylaxis in guinea pigs. Based on a comparison of measured body weights, there were no changes in body weight for the KRGM treatment group compared with the control group. In the ovalbumin treated group, however, there was a statistically significant decrease in body weight. For the active systemic anaphylaxis test, after the induction, there were no symptoms that suggested anaphylactic shock in the control and KRGM treatment group. In the ovalbumin treated group, there were symptoms that suggested severe anaphylaxis, and those symptoms included restlessness, piloerection, tremor, rubbing or licking the nose, sneezing, coughing, hyperpnea, dyspnea, staggering gait, jumping, gasping and writhing, convulsion, side position and Cheyne-stokes respiration. All animals died within thirty minutes in the ovalbumin treated group. For the passive cutaneous anaphylaxis test in guinea pigs sensitized to KRGM, each anti-serum was diluted in a stepwise manner. This was followed by an intravenous injection of a mixture of KRGM and Evans blue. The results of the test showed that all the responses were negative in the control and the low-dose and high-dose administration groups. However, in the ovalbumin treated group, all the responses were positive. Based on the above results, there were no anaphylactic responses for up to 12 times the amount of human intake of KRGM in Hartley Guinea-pigs. The results suggest that KRGM is safe as measured by the systemic and local antigenicity in guinea pigs.

Ginseng has been traditionally used for several millennia in Asian countries, including Korea, China, and Japan, not only as a nourishing and tonifying agent but also as a therapeutic agent for a variety of diseases. In recent years, the various effects of red ginseng including immunity improvement, fatigue relief, memory improvement, blood circulation improvement, antioxidation, mitigation of menopausal women's symptoms, and anticancer an effect have been reported in clinical as well as basic research. Around the world, there is a trend of the rising consumption of health functional foods on the level of disease prevention along with increased interest in maintaining health because of population aging and the awareness of lifestyle diseases and chronic diseases. Red ginseng occupies an important position as a health functional food. But till now, international ginseng monographs including those of the World Health Organization have been based on data on white ginseng and have mentioned red ginseng only partly. Therefore, the red ginseng monograph is needed for component of red ginseng, functionality certified as a health functional food in the Korea Food and Drug Administration, major efficacy, action mechanism, and safety. The present red ginseng monograph will contribute to providing accurate information on red ginseng to agencies, businesses, and consumers both in South Korea and abroad. Keywords: Health functional foods, Red ginseng, Red ginseng monograph

Elevated plasma LDL cholesterol is the dominant risk factor for the development of atherosclerosis and cardiovascular disease. Deficiency in the LDL receptor (LDLR) is a major cause of familial hypercholesterolemia in humans, and the LDLR knockout mouse is a major animal model of atherosclerosis. Here we report the generation and characterization of an ldlr mutant zebrafish as a new animal model to study hypercholesterolemia and vascular lipid accumulation, an early event in the development of human atherosclerosis. The ldlr mutant zebrafish were characterized by activated SREBP-2 pathway and developed moderate hypercholesterolemia when fed a normal diet. However, a short-term, 5-day feeding of ldlr mutant larvae with a high-cholesterol diet (HCD) resulted in exacerbated hypercholesterolemia and accumulation of vascular lipid deposits. Lomitapide, an inhibitor of apoB lipoprotein secretion, but not the antioxidant probucol, significantly reduced accumulation of vascular lipid deposits in HCD-fed ldlr mutant larvae. Furthermore, ldlr mutants were defective in hepatic clearance of lipopolysaccharides, resulting in reduced survival. Taken together, our data suggest that the ldlr knockout zebra­fish is a versatile model for studying the function of the LDL receptor, hypercholesterolemia, and related vascular pathology in the context of early atherosclerosis.—

Retinal apoptosis plays a critical role in the progression of diabetic retinopathy (DR), a common diabetic complication. Currently, the tight control of blood glucose levels is the standard approach to prevent or delay the progression of DR. However, prevalence of DR among diabetic patients remains high. Focusing on natural nutrients or herbal medicines that can prevent or delay the onset of diabetic complications, we administered an ethanol extract of the aerial portion of Osteomeles schwerinae (OSSCE), a Chinese herbal medicine, over a period of 17 weeks to spontaneously diabetic Torii (SDT) rats. OSSCE was found to ameliorate retinal apoptosis through the regulation of advanced glycation end product (AGE) accumulation, oxidative stress, and mitochondrial function via the inhibition of NF-&kappa, B activity, in turn, through the downregulation of PKC&delta, P47phox, and ERK1/2. We further demonstrated in 25 mM glucose-treated human retinal microvascular endothelial cells (HRMECs) that hyperoside (3-O-galactoside-quercetin), quercitrin (3-O-rhamnoside-quercetin), and 2&Prime, O-acetylvitexin (8-C-(2&Prime, O-acetyl-glucoside)-apigenin) were the active components of OSSCE that mediated its pharmacological action. Our results provide evidence that OSSCE is a powerful agent that may directly mediate a delay in the development or disease improvement in patients of DR.

Background Osteomeles schwerinae C. K. Schneid. (Rosaceae, OSSC) is a medicinal plant traditionally used to treat various diseases in Asia. The chemical constituents of OSSC have an inhibitory effect on aldose reductase activity, which has been implicated in the pathogenesis of diabetic complications. However, the protective effects of the pharmacological activity and potential mechanisms in diabetic nephropathy are still not known. Objective In the present study, OSSC extracts and major compounds were examined for their effects on binding to the receptors of advanced glycation end products (RAGE) and on transforming growth factor-beta1 (TGF-β1) expression-related signal mechanisms in mouse glomerular mesangial cells (GMCs). Materials and methods A simple, rapid and efficient method was developed for the simultaneous determination of the marker compounds in the ethanol extract of the leaves and twigs of OSSC using HPLC-diode array detector (DAD). In this study, we determined the effects of OSSC extract and hyperoside on AGE and RAGE binding, and studied the mechanism of OSSC extract effects on AGE-bovine serum albumin (BSA)-treated GMCs. GMCs overexpressing human RAGE were cultured in AGE-BSA labeled with Alexa 488, and OSSC extract. AGE/RAGE binding were measured using fluorescence (excitation 485 nm/emission 528 nm). TGF-β1 protein expression levels were determined by western blot analyses. Results OSSC extracts of leaves and twigs inhibited on AGE/RAGE binding and TGF-β1 protein expression in a dose-dependent manner in GMCs. Furthermore, OSSC extracts reduced the effects on AGE-BSA-induced reactive oxidative species (ROS) formation and nuclear translocalization of transcription factor NF-κB. OSSC extracts inhibited phosphorylation of extracellular signal-regulated protein kinases1/2 (ERK1/2), p38 mitogen-activated protein kinases (p38MAPK), and IκB. Hyperoside also inhibited AGE/RAGE binding and ROS formation, and reduced TGF-β1 expression and IkB phosphorylation. Conclusions OSSC extracts and hyperoside may attenuate AGE/RAGE binding and expression of TGF-β1 by downregulating of pERK1/2, p38MAPK and IκB phosphorylations in GMCs under diabetic condition and retard the development of diabetic complications such as diabetic nephropathy.

Objectives: The purpose of the study is to investigate the influence of community oral hygiene program on oral hygiene practice behavior in children. Methods: Oral hygiene promotion program was performed in 23 community child centers in Seoul, Incheon and Gyeonggido. The study analyzed the effect of community oral hygiene promotion programs on the oral hygiene practice behavior in children from September to December, 2014. The trained dental hygienists in 23 public health centers and dental hygiene students participated in the oral hygiene promotion program for oral health examination and education for the children. The contents of the program was standardized and instructed to the team members. The individual improvement in children oral hygiene practice was assessed using PHP(patient hygiene performance) index score. Data were analyzed using SPSS 12.0 program. Results: After the program, children who stopped eating confectioneries and those eating once a day increased to 32.12% and 14.24%, respectively. Those eating more than four times a day were still high(32.44%), but it was a lower rate than before the program (p

Background and Purpose Although a variety of animal models have been used to test drug candidates and examine the pathogenesis of diabetic retinopathy, time-saving and inexpensive models are still needed to evaluate the increasing number of therapeutic approaches. Experimental Approach We developed a model for diabetic retinopathy using the early stage of transgenic zebrafish (flk:EGFP) by treating embryos with 130 mM glucose, from 3-6 days post fertilisation (high-glucose model). On day 6, lenses from zebrafish larvae were isolated and treated with 3% trypsin, and changes in hyaloid-retinal vessels were analysed using fluorescent stereomicroscopy. In addition, expression of tight junction proteins (such as zonula occludens-1), effects of hyperosmolar solutions and of hypoxia, and Vegf expression were assessed by RT –PCR. NO production was assessed with a fluorescent substrate. Effects of inhibitors of the VEGF receptor, NO synthesis and a VEGF antibody (ranibizumab) were also measured. Key Results In this high-glucose model, dilation of hyaloid-retinal vessels, on day 6, was accompanied by morphological lesions with disruption of tight junction proteins, overproduction of Vegf mRNA and increased NO production. Treatment of this high-glucose model with an inhibitor of VEGF receptor tyrosine kinase or an inhibitor of NO synthase or ranibizumab decreased dilation of hyaloid-retinal vessels. Conclusions and Implications These findings suggest that short-term exposure of zebrafish larvae to high-glucose conditions could be used for screening and drug discovery for diabetic retinopathy and particularly for disorders of retinal vessels related to disruption of tight junction proteins and excessive VEGF and NO production.

Over half of anti-NMDA receptor encephalitis (NMDARE) is unrelated to etiologies such as teratomas or herpes simplex encephalitis. Bacillus mannanilyticus nonribosomal peptide synthetase (NRPS) shares a protein sequence with GluN1. After confirming the anti-NRPS antibody immunoreactivity in an index patient by liquid chromatography-tandem mass spectrometry (LC-MS/MS), we identified 24/57 (42%) patients with similar immunoreactivity patterns by western blotting. These patients mostly (20/24 [83%]) did not have ovarian teratomas, had fewer medial temporal (2/24 [8%]) and any (5/24 [21%]) brain lesions and less pleocytosis (13/24 [54%]). These results identified an anti-NMDARE subgroup with a distinct immunoreactivity pattern, which needs further investigation.

Background and Purpose The main difficulty when diagnosing leptomeningeal metastases (LMSs) is the low sensitivity of cytology. Cancer cells release cell-free DNA (cfDNA) during proliferation and apoptosis, and so we analyzed the cfDNA level as a biomarker for LMSs in hematologic malignancy. Methods This study prospectively enrolled 20 patients with hematologic malignancy who underwent cerebrospinal fluid (CSF) analysis. LMS was diagnosed based on both CSF cytology and clinical findings. Results The CSF level of cfDNA was higher in patients with LMSs (108.17±84.84 ng/mL, mean±standard deviation) than in non-LMS patients (14.23±2.78 ng/mL). The sensitivity of cfDNA was higher than that of cytology (100% vs. 87%). Conclusions The cfDNA level in the CSF can be used as a supplemental marker for diagnosing LMS in hematologic malignancy patients.

Background:Panax ginseng C.A. Meyer is one of the most frequently used herbs in the world. The roots of Panax ginseng have been used as a traditional tonic and medicine for thousands of years in Korea and China. Today, ginseng root is used as a dietary supplement and complementary medicine and for adjuvant therapeutics worldwide. The efficacy of ginseng has been studied in a wide range of basic research and clinical studies. However, it has been reported that the results from clinical studies are conflicting, and they depend on the parameters of the protocol design including the conditions of the participants and the types of ginseng used such as red ginseng, white ginseng, fermented ginseng and cultured ginseng. [...].

The increasing cross-border mobility of dental school or dental hygiene students, educators, practitioners, programs and providers takes challenges for existing national quality assurance and accreditation frameworks and bodies, as well as for the systems for recognizing foreign qualifications. The new dental hygiene accreditation system was introduced to encourage the improvement of dental hygiene programs, to ensure the quality of education and, most of all, to establish an internationally compatible system of evaluation and accreditation. The accreditation procedure takes 1 year to complete. The result of the accreditation is released after evaluation via self-study report, site visit, preliminary draft report, responses from the institution and the results from the conciliation and review committees. The result from the accreditation procedur e is either ‘accreditation’ or ‘no accreditation’. Accredited schools receive one of several statuses following the evaluation. These are next general review, interim report and interim visit or su spension. Dental healthcare quality is not improved instantaneously, but instead gradually through continuous communication within the dental field. For this accreditation system to be successful, the following are essential: the accreditation agency should adop t hygiene education accreditation; it needs to become financially independent and managed efficiently; the autonomy and regulations surrounding the system need to be balanced; the professionalism of the system is ensured; and the dental field which includes not only dental program, but also hygiene program, needs to play an active role in the operation of the system.

Accumulating evidence suggests that a distinct subpopulation of cancer stem cells (CSCs) is responsible for tumour initiation and progression in head and neck squamous cell carcinoma (HNSCC). Wnt/β-catenin signalling is essential for stem cell regulation and tumourigenesis, but its molecular mechanism in HNSCC CSCs remains unknown. We investigated whether Wnt/β-catenin signalling regulates self-renewal and tumourigenicity of HNSCC stem-like cells in vitro and in vivo. Cytoplasmic/nuclear β-catenin, a major effector of Wnt/β-catenin signalling, was expressed in a subpopulation of tumour cells in primary HNSCC tissue but in none of normal head and neck tissues. Overexpression of β-catenin increased proliferation of HNSCC cells and induced dedifferentiation of these cells to cells with stem-like features. Knockdown of β-catenin in HNSCC stem-like cells blocked their self-renewal capacity, stemness-associated gene expression, chemoresistance, and in vivo tumourigenicity. Furthermore, β-catenin directly regulates Oct4 transcription in HNSCC stem-like cells. In addition, the effect of shRNA-mediated repression of β-catenin on CSC traits in HNSCC stem-like cells was reversed by overexpression of Oct4. In patients with HNSCC, higher levels of both cytoplasmic/nuclear β-catenin and Oct4 correlated with the worst prognosis. These results suggest inhibition of Wnt/β-catenin signalling as a novel therapeutic strategy for targeting HNSCC stem-like cells.

Objectives : The purpose of this study was to examine the oral health programs for children of the low income classes in order to prevent oral health problems and to provide the effective oral health programs in community children`s centers. Methods : The subjects were 464 teachers in community children`s centers in Seoul, Incheon, Gyeonggi-do, Chungcheongbuk-do, and Jeollabuk-do. Results : There were differences between the regions(p

SUMMARY In difference in differences, a treatment is applied only to a qualified group at some time point. The qualification may be time-constant as in gender, or time-varying as in residential location. When the qualification is time-varying, there appear four groups: the newly qualified (in-movers), the already qualified (in-stayers), the newly disqualified (out-movers), and the already disqualified (out-stayers). A change in qualification may affect the response variable of interest even when the treatment effect is zero, which is an ‘untreated moving effect’. Also, when the treatment effect is not zero, it may be different across the four groups. The conventional difference in differences fails to remove untreated moving effects and ignores the possible treatment effect heterogeneity across the groups. This paper shows how to account for untreated moving effects and proposes ‘the effect on in-stayers’ as the main effect of interest. Our proposal can be implemented with least squares estimator for panel models or with nonparametric methods. An empirical analysis is provided using Korean data for the effects of the basic elder pension on health-care expenditure. Copyright © 2014 John Wiley & Sons, Ltd.

The purpose of this study was to identify the effects of Qigong training of elderly single women on their pain consciousness and depression. To achieve the goal of this study, 102 female elder`s were randomly placed into three groups of Health Qigong such as YiJinJing training group, LiuZiJue training group and control group with no training. They were trained for 12 weeks and the changes in their pain consciousness and depression were observed. After identifying the difference among three groups, the following conclusions were made. First, pain consciousness and depression were significantly improved in YiJinJing training group and LiuZiJue training group in pre and post assessments, while there was no difference in the control group. Second, in one-way ANOVA analysis of three groups on pain consciousness, there was significant difference between control group and YiJinJing training group, between control group and LiuZiJue training group, and between YiJinJing training group and LiuZiJue training group. Third, in ANOVA analysis of three groups on depression, there was significant difference between control group and YiJinJing training group, and between control group and LiuZiJue training group. The results show that YiJinJing training and LiuZiJue training of Health Qigong made effects on pain consciousness and depression of elderly single women, in particular YiJinJing training was more effective than LiuZiJue on pain consciousness.

The anti-proliferative activity of β-ionone was investigated on human non-small lung cancer A-549 cells (designated A-549 cells). A-549 cells were treated with various concentrations of β-ionone (1, 5, 10, and 15 μM) for two, four, and six days. Biochemical markers related to the growth inhibition of A-549 cells by β-ionone were measured at the second day of incubation. β-Ionone inhibited the growth of A-549 cells by dose-and time-dependent manners, resulting in an IC 50 of 5.0 μg/ml at the second day of incubation. β-Ionone induced apoptosis by a dose-dependent manner. β-Ionone increased levels of p53, p21, and Bax proteins, but suppressed expression of the Bcl-2 protein. Similarly, β-ionone enhanced cytochrome c release from the mitochondria to the cytosol, and induced activation of caspase-9 and -3. Additionally, β-ion-one reduced cPLA₂ and COX-2 protein levels. These results suggest that the β-ionone inhibits the proliferation of A-549 cells through reciprocal regulation of Bax and Bcl-2 gene expression and suppression of cPLA₂ and COX-2 protein expressions.

The use of a microorganism, or its secretions, to prevent plant disease offers an attractive alternative or supplement to synthetic fungicides for the management of plant disease without the negative effects of chemical control mechanisms. During a screening for microorganisms with the potential to be used as microbial fungicides, Bacillus sp. BS061 was isolated from a plant leaf. The strain BS061 potently inhibited the mycelial growth of Botrytis cinerea, and significantly reduced disease incidence of powdery mildew in cucumber and strawberry. We also found that the culture filtrate of BS061 inhibited the mycelial growth of various plant pathogens.

CD44 is generally accepted as a surrogate marker for head and neck squamous carcinoma cancer stem cells (HNSC CSCs) and only CD44+ HNSC cells have tumour initiating capacity. However, a recent report suggested that CSCs themselves might be heterogenous due to various genetic alterations. Here, we compared in vitro stem-like cell characteristics, chemoresistance and in vivo tumour formation capacity of CD44+ and CD44- HNSC cells obtained from primary HNSC patient specimens. CD44- HNSC spheroid cells generated spheroid cells again after seeding of single-dissociated spheroid CD44- HNSC cells. Immunocytochemistry assays revealed that various stem cell markers, including octamer-binding transcription factor 4 (OCT4), sex determining region Y-box 2 (SOX2) and nestin were up-regulated in CD44- spheroid cells, similar to CD44+ spheroid cells. Furthermore, CD44- spheroid cells appeared to be chemoresistant to cisplatin and showed increased levels of ABCG2, similar to CD44+ spheroid cells. Of most interest, as few as 1000 CD44- spheroid cells were able to give rise to tumours in nude mice. The collective data indicate that the cell surface marker CD44 cannot be used as a selective marker of spheroid-forming, tumour-initiating or chemoresistant cell populations, and further indicate the limitation of current HNSC CSC identification methods using the CD44 cell surface marker.

c-Met proto-oncogene, which is a receptor of ligand hepatocyte growth factor (HGF), has been associated with cancer cell invasion. There have been no reports on the relationship between the expression of c-Met and tumor invasion and metastasis in small (T(1-2)) squamous cell carcinoma of the oral tongue (SCCOT). We analyzed the relationship between c-Met expression and tumor invasion depth and lymph node metastasis in 71 surgically treated patients with small SCCOT using immunohistochemistry. Furthermore, we investigated the associations between the c-Met expression status and patient survival. In addition, we explored whether overexpression of c-Met enhances tumor growth and invasion of tongue cancer cells in vitro and in vivo. Positive immunohistochemical staining of c-Met was observed in 39 (55%) samples. Presence of neck metastasis, and >4mm depth of tumor invasion, strongly correlated with c-Met expression in small SCCOT both by the univariate and multivariate analysis (p

The aim of this study was to compare serum and cerebrospinal fluid (CSF) cytokine/chemokine levels between anti-NMDAR and anti-LGI1 encephalitis patients. Samples from fourteen anti-NMDAR encephalitis patients, ten anti-LGI1 encephalitis patients, and ten controls were analyzed for the following cytokines/chemokines: IL-1b, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12, IL-13, IL-17A, IL-23, GM-CSF, IFN-gamma, TNF-alpha, and CXCL13. Compared with controls, CSF IL-17A, IL-6 and CXCL13 were elevated in anti-NMDAR encephalitis patients (post-hoc p-values 0.002, 0.011, and 0.011, respectively) but not in anti-LGI1 encephalitis patients. In the serum, only IL-2 was increased in anti-NMDAR encephalitis. Intrathecal IL-17/IL-6 activation is a characteristic of anti-NMDAR encephalitis.

SUMMARY If informal family health care is a substitute for formal health care, then there is a scope to reduce formal health care cost by promoting informal family health care. With the use of Korean data for the elderly, this paper estimates the effects of informal family health care on formal health care, where the former is measured by the number of caregivers and the latter is measured by the formal health care expenditure. This task, however, poses a number of difficulties. The first is that the number of the family caregivers is an endogenous count regressor. The second is that there are too many zeros in the count (85%). The third is that the response variable also has a nontrivial proportion of zeros (14%). This paper overcomes these problems by combining ‘control function approach’, ‘zero-inflated’ counts, and a semiparametric estimator for censored models. The resulting procedure avoids strong parametric assumptions and behaves well computationally. Our main empirical finding is that informal family health care has a large substitute effect for diabetics and that there are also weak evidences that informal family health care has substitute effects for high blood pressure and mental diseases. Copyright © 2012 John Wiley & Sons, Ltd.

BACKGROUND: Ginseng saponin and ginsenosides exert anti-obesity effects via the modulation of physiological lipid metabolism in vivo or intracellular signalling in cell culture systems. However, the complicated relationship between the anti-obesity effects of ginseng and gene expression has yet to be defined under in vivo conditions. Therefore, we evaluated the relationship between the anti-obesity effects of Korean red ginseng extract (KRGE) and hepatic gene expression profiles in mice fed long-term on a high-fat diet (HFD) in this study. RESULTS: KRGE reduces the levels of cholesterol, low-density lipoprotein-cholesterol (LDL-C), serum triglycerides, and atherogenic indices. Levels of leptin, adiponectin and insulin, which regulate glucose and lipid metabolism, were impaired profoundly by HFD. However, KRGE treatment brought these levels back to normal. KRGE was found to down-regulate genes associated with lipid metabolism or cholesterol metabolism (Lipa, Cyp7a1, Il1rn, Acot2, Mogat1, Osbpl3, Asah3l, Insig1, Anxa2, Vldlr, Hmgcs1, Sytl4, Plscr4, Pla2g4e, Slc27a3, Enpp6), all of which were up-regulated by HFD. CONCLUSION: KRGE regulated the expression of genes associated with abnormal physiology via HFD. Leptin, insulin, and adiponectin, which carry out critical functions in energy and lipid metabolism, were shown to be modulated by KRGE. These results show that KRGE is effective in preventing obesity. Copyright © 2011 Society of Chemical Industry

Purpose: This study was to compare the influencing factors of clinical competence between associate and bachelor nursing students completing nursing courses. Method: The subjects were 429 students completing the nursing courses (172 bachelor course students, and 257 associate course students). The study design was cross sectional. Results: Bachelor students showed a significant higher score in clinical competence than associate students. There were significant influencing factors on clinical competence including critical thinking disposition, professional conduct, communication skill, theoretical education satisfaction, and practice education satisfaction. The strongest factor influencing clinical competence was critical thinking disposition. All the factors mentioned above explained 49% of clinical competence in associate courses, and 40.0% in bachelor courses excluding the factor of practice education satisfaction. The level of theoretical and practice education satisfaction of bachelor courses was significantly lower than associate. Conclusion: It is recommended that associate course nursing students should be encouraged in critical thinking disposition, professional conduct, and communication skill. Bachelor course nursing students should be encouraged in practical education including practical environments, instruction methods, and content for improving clinical competence.

This study aimed to identify success factors of smoking cessation for 6 months in Adolescent smokers. Data were collected from January to December, 2008. The 925 adolescent smokers were registered smoking cessation clinic at public health center in Incheon. Data(demographic, smoking, smoking cessation) were analyzed by logistic regression. Major factors are attempting to quit smoking(no trial, OR

In this study, the inhibitory effects of quercitrin gallate (QG), a polyphenolic compound, were investigated on aldose reductase (AR) activity as well as on antioxidant levels and enzyme activity in the lens. The effects of QG on xylose-induced lens opacity (cataracts), as seen in diabetes, were also examined. QG showed potential inhibitory activity against rat lens AR activity with an IC50 value of 0.064 μM. The xylose-induced opacity of the lens was significantly improved after treatment with QG. Exposure of rat lenses to xylose significantly increased the oxidized glutathione/reduced glutathione ratio, and treatment with QG significantly decreased this ratio. Antioxidant enzyme (i.e., superoxide dismutase and catalase) activities were significantly decreased under high xylose conditions, and treatment with QG significantly increased the activities and levels of these factors. These results suggest that QG may provide a potential therapeutic approach for the prevention of diabetic complications, such as cataracts.

In this study, the anticoagulant compounds in Korean red ginseng (KRG) were investigated. KRG powder was extracted using hot methanol, and the methanol extract was fractionated into n-hexane, ethylacetate, n-butanol, and aqueous fractions by solvent partitioning. The remains from the methanol extraction were further extracted with water and then dialyzed to obtain low and high molecular weight fractions. The anticoagulant activities of the seven fractions were evaluated in terms of thrombin time, prothrombin time, and activated partial thromboplastin time. Among these fractions, the ethylacetate fraction showed the most potent anticoagulant activity. The active components in the ethylacetate fraction were identified as the phenolic compounds vanillic, caffeic, ferulic, and p-coumaric acid via TLC and HPLC. These findings suggest that the anticoagulant activities of phenolic compounds contribute to the cardiovascular effects of KRG.

Non-enzymatic glycation reactions between reducing sugar and free reactive amino groups of protein lead to the formation of advanced glycation end products, which increase under conditions of aging or diabetes. A previous study showed that extracts of Cassiae Semen (CS), the seed of Cassia tora, had inhibitory activity on advanced glycation end products formation in vitro. To examine the pharmacological effects of a butanol-soluble extract of CS under conditions of diabetic nephropathy, we evaluated the expression of transforming growth factor-beta1 (TGF-beta1) and fibronectin, key mediators of diabetic nephropathy, in mouse glomerular mesangial cells cultured in the presence of S100b (a specific ligand for receptor of advanced glycation end products). CS inhibited S100b-induced TGF-beta1 and fibronectin expression in mouse mesangial cells by suppressing activation of Smad2/3, extracellular signal-regulated kinase (ERK)/mitogen-activated protein kinase (MAPK), and oxidative stress. Moreover, CS suppressed nuclear factor-kappa B (NF-kappaB) activation in S100b-stimulated mouse mesangial cells. To identify the active compounds of CS, three major compounds, rubrofusarin-6-O-beta-d-gentiobioside (CS-A), toralactone-9-O-beta-d-gentiobioside (CS-B), and cassiaside (CS-C), were tested in cells. Of these compounds, CS-A significantly decreased the expression of TGF-beta1 and fibronectin and NF-kappaB DNA binding activity. These findings suggest that CS, especially CS-A, has potential as a preventive agent for advanced glycation end products-related diabetic complications.

Aim of the study To investigate the effects of KIOM-79 in preventing the development of diabetic complications, such as cataracts. Materials and methods The inhibitory effects of KIOM-79 were assessed in a model of xylose-induced lens opacity and on changes mediated by high levels of glucose in human lens epithelial (HLE-B3) cells. Results In lenses treated with KIOM-79, opacity was significantly improved and glutathione (GSH) was increased compared to controls. In HLE-B3 cells treated with KIOM-79, high glucose-mediated increases in TGF-β2, αB-crystallin, and fibronectin were significantly inhibited in a dose-dependent manner. KIOM-79 decreased the phosphorylation of p-Smad2/3, pp38MAPK, pp44/42, and NF-κB signaling in cells grown under high glucose conditions. Conclusion KIOM-79 is protective against lens opacity and protects HLE-B3 cells from the toxic effects of high glucose. Therefore, KIOM-79 may provide a potential therapeutic approach for preventing diabetic complications, such as cataracts.

High sugar levels found in diabetic cataract cause the opacification of lenses by osmotic changes induced via the aldose reductase (AR)-mediated polyol pathway. In this study, puerariafuran, a 2-arylbenzofuran from Pueraria lobata, investigated the inhibitory effects upon AR, antioxidant contents and enzyme activities in the lens. The effect of puerariafuran on xylose-induced lens opacity was also examined. Puerariafuran showed potential inhibitory activity with an IC50 value of 22.34 microM against rat lens AR. The xylose-induced opacity of lenses was significantly improved when treated with puerariafuran. Xylose exposure of rat lenses significantly decreased the reduced glutathione (GSH)/oxidized glutathione (GSSG) ratio, superoxide dismutase (SOD), and catalase (CAT) activity and treatment with puerariafuran significantly increased these factors. These results suggest that puerariafuran may provide a potential therapeutic approach for prevention of diabetic complications, such as cataracts.

Plant-based vaccines have been produced in transgenic plants including tobacco, potatoes, corn, and rice. However, these plants are not suitable for administration without cooking. To overcome this obstacle, a fusion gene encoding the synthetic enterotoxigenic Escherichia coli heat-labile enterotoxin B subunit genetically fused with a synthetic neutralizing epitope of porcine epidemic diarrhea virus (sLTB-sCOE) was introduced into lettuce cells (Lactuca sativa) by Agrobacterium-mediated transformation methods. The integration and expression of the sLTB-sCOE fusion gene was confirmed in transgenic lettuce by genomic DNA PCR amplification and Northern blot analysis, respectively. Synthesis and assembly of the LTB-COE fusion protein into oligomeric structures with pentamer size were observed in transgenic plant extracts by Western blot analysis with anti-LTB or anti-COE antibodies. The binding of plantproduced LTB-COE to intestinal epithelial cell membrane glycolipid receptors was confirmed by GM1-ganglioside enzyme-linked immunosorbent assay (GM1-ELISA). Based on the ELISA results, LTB-COE fusion protein made up about 0.026∼0.048% of the total soluble protein in the transgenic lettuce leaf tissues. The synthesis and assembly of LTB-COE monomers into biologically active oligomers in transgenic lettuce leaf tissues demonstrates the feasibility of using uncooked edible plant-based vaccines for mucosal immunization.

Aim of the study In this study, we investigated whether KIOM-79 inhibits transforming growth factor-beta 1 (TGF-β1) and fibronectin expression in mouse mesangial cells cultured under S100b, a specific ligand of the receptor for advanced glycation end products (RAGE). Materials and methods Cell counting kit (CCK-8) assay was employed to evaluate the viability of KIOM-79-treated mesangial cells. The effect of KIOM-79 on S100b-induced TGF-β1 and fibronectin expression was investigated using RT-PCR, ELISA, and Western blot on mesangial cells. Results KIOM-79 (up to 50 μg/ml) appeared to have no effect on cell viability. S100b induced an increase in the expression TGF-β1 and fibronectin. Expression of TGF-β1 and fibronectin was inhibited significantly by KIOM-79 treatment in mesangial cells. KIOM-79 also inhibited the expression of NF-kB and inactivated p38 mitogen-activated protein kinase (MAPK) and extracellular signal-regulated kinase (ERK) 1/2 in mesangial cells. KIOM-79 pretreatment inhibited increased malondialdehyde (a product of lipid peroxidation and a marker for oxidative stress) levels in S100b-induced mesangial cells. Conclusions These data demonstrate that KIOM-79 inhibits expression of TGF-β1 and fibronectin through inactivation of MAPK/ERK1/2 signaling, reduction in malondiadehyde levels, and inhibition of NF-kB in mesangial cells cultured under diabetic conditions. KIOM-79 could be beneficial for preventing of the development of diabetic complications such as nephropathy.

Advanced glycation end products (AGEs) have been implicated in diabetic complications. To measure AGEs, especially Nɛ-(carboxymethyl)lysine (CML), in sera from Zucker diabetic fatty rats (ZDF) and Zucker lean rats (ZL), we used a novel method of protein chip and surface plasmon resonance imaging (SPRI). Serum samples were obtained from male ZDF and ZL rats at 20 weeks of age. Antibodies to AGEs or CML were immobilized on a gold surface, which was modified by cysteine-tagged, protein-G constructs. The gold chip upon which the serum was spotted was optically coupled with a prism coupler. The reflected images from the gold chip were obtained using a charge-coupled device (CCD) camera. The direct analysis of the glycated proteins and products using SPRI showed that AGEs and CML levels were elevated in ZDF serum, compared with ZL serum. The lowest detection limit of AGEs was 10 ng/ml, with a working range covering the physiological range. These results indicate that the protein chip and SPRI system is very suitable for the measurement of glycated proteins and end products in serum samples. This system offers high sensitivity without any fluorescent or other labeling of the components and saves a substantial amount of time, resources, and labor. Our results suggest that SPRI systems can be used as a tool to diagnose diabetic complications.

Hyperglycemia-induced oxidative stress has been suggested as a mechanism underlying diabetic complications. Oxidative stress triggers cell death in various cell types, including glomerular mesangial cells which play important roles in diabetic nephropathy. In the present study, we investigated the potential cytoprotective effect of erigeroflavanone, a novel flavanone derivative from the flowers of Erigeron annuus, in cultured mouse mesangial cells using hydrogen peroxide (H2O2) as an oxidative stress inducer. Our data show that hydrogen peroxide induced a decrease in cell viability that was attenuated by erigeroflavanone. Hydrogen peroxide treatment increased formation of dichlorofluorescein (DCF)-sensitive intracellular reactive oxygen species (ROS). This enhanced ROS formation was significantly reduced by pretreatment with erigeroflavanone in a dose-dependent manner. Hydrogen peroxide treatment also induced phosphorylation of the mitogen-activated protein kinases (MAPKs), c-Jun terminal kinase (JNK), extracellular-regulated kinase (ERK) and p38, and activated caspase-3. Pretreatment with erigeroflavanone inhibited hydrogen peroxide-induced activation of MAPKs and caspase-3. From these data we conclude that erigeroflavanone provides a protective effect against oxidative stress-induced cell death in mesangial cells that is associated with its antioxidant action and inhibition of MAPKs and caspase-3. These results suggest that erigeroflavanone has potential as a therapeutic agent in the treatment of renal diabetic complications.

The goal of this study is to analyze male college students' awareness and behaviors toward appearance management, and expenditure for appearance management. Male subjects' appearance management behaviors were focused on hair, skin, cosmetic surgery, fashion and body shape. A chi-square test, t-test, one-way ANOVA, Pearson's correlation analysis, multiple regression analysis were used. The major findings were as follows: (1) The mean of male college students' 'awareness toward appearance management' is 30.01(out of a possible 50). (2) The variables that significantly influenced their awareness toward appearance management are 'an allowance amount per month' and 'household income'. (3) Subjects' most preferred behavior is a variable of 'fashion care' in the appearance management(29.9%). (4) Appearance management behavior is different, according to demographic variables and awareness toward appearance management. (5) The mean of 'monthly expenditure for appearance management' is 247,194won. Consequently, the study shows that there is a most significant relation of 'awareness toward appearance management' to 'expenditure for appearance management'

Aim of the study KIOM-79 retards the development of diabetic nephropathy in animal models of type 1 and type 2 diabetes. In this study, we evaluated whether KIOM-79 could prevent apoptotic cell death and advanced glycation end products (AGEs) accumulation in the retinas of diabetic db/db mice. Material and methods Mice were treated orally with KIOM-79 (150 mg/kg body weight) once daily for 12 weeks. Levels of retinal ganglion cell death were measured by terminal dUTP nick-end labeling (TUNEL) assay. In the retina, the activity of caspase-3 (a marker of apoptosis) and the formation of AGEs were measured by immunohistochemical staining. Results KIOM-79 reduced the number of TUNEL-immunoreactive retinal cells. KIOM-79 attenuated caspase-3 expression and AGEs accumulation in the retina. Conclusions These data show that KIOM-79 can prevent apoptosis in neuronal cells, AGEs accumulation in the retina, and retard the development of diabetic retinopathy.

We expressed the cholera toxin B subunit (CTB) fused to an endoplasmic reticulum retention signal (SEKDEL) in carrot roots using an Agrobacterium-mediated transformation method. Fourteen independent transgenic lines were regenerated via somatic embryogenesis after 6 months of culture. The sCTB gene was detected in the genomic DNA of transgenic carrot by PCR amplification. Expressions and assembly of sCTB protein into oligomeric structures of pentameric size were observed in transgenic plant extracts by Western blot analysis. The sCTB produced by transgenic carrot roots demonstrated strong affinity for GM1-ganglioside, suggesting that the sCTB conserved the antigenic sites for binding and proper folding of the pentameric sCTB structure. The expression level of sCTB comprised approximately 0.48% of total soluble protein (TSP) in root of transgenic carrot.

Compound K (CK), a protopanaxadiol ginsenoside metabolite, was previously shown to have immunomodulatory effects. In this study, we isolated the CK rich fractions (CKRF) from Korean Red Ginseng and investigated the regulation of CKRF-mediated inflammatory signaling during Toll-like receptor (TLR)-mediated cellular activation. Among various TLR ligands, CKRF considerably abrogated TLR4- or TLR9-induced inflammatory signaling. Both LPS and CpG-containing oligodeoxynucleotides (CpG-ODN) stimulation rapidly activates mitogen-activated protein kinases [MAPKs; extracellular signal-regulated kinases 1/2 and p38], NF-, and expression of pro-inflammatory cytokines tumor necrosis factor-, and interleukin-6 in murine bone marrow-derived macrophages (BMDMs) in a time- and dose-dependent manner. Of interest, pre-treatment of CKRF in either LPS/TLR4- or CpG-ODN/TLR9-stimulated macrophages substantially attenuated the LPS-induced inflammatory cytokine production and mRNA expressions, as well as MAPK and NF- activation. To our knowledge, this is the first description of the inhibitory roles for CKRF in TLR4- or TLR9-associated signaling in BMDMs. Collectively, these results demonstrate that CKRF specifically modulates distinct TLR4 and TLR9-mediated inflammatory responses, and further studies are urgently needed for their in vivo roles for potential therapeutic uses, such as in systemic inflammatory syndromes.