AIM: To investigate p53 mutation and p21 expression in hepatocarcinogenesis induced by hepatitis B virus (HBV) and aflatoxin B1 (AFB1) in tree shrews, and to reveal the role of these genes in hepatocarcinogenesis.METH...AIM: To investigate p53 mutation and p21 expression in hepatocarcinogenesis induced by hepatitis B virus (HBV) and aflatoxin B1 (AFB1) in tree shrews, and to reveal the role of these genes in hepatocarcinogenesis.METHODS: Tree shrews were divided into four groups:group A, those infected with HBV and fed with AFB1 (n = 39);group B, those infected with HBV alone (n = 28); group C,those fed with AFB1 alone (n = 29); and group D, normal controls (n = 20). The tree shrews underwent liver biopsies once every 15 wk. Expression of p53 and p21 proteins and genes in the biopsies and tumor tissues of the experimental tree shrews was detected, respectively, by immunohistochemistry,and by Southem blotting and reverse transcription-polymerase chain reaction and sequencing.RESULTS: The incidence of hepatocellular carcinomas (HCC) was higher in group A (66.7%) than that in group B (3.57%) and C (30%). The time of HCC occurrence was also earlier in group A than that in group C (120.0±16.6 wk vs 153.3±5.8 wk, respectively, P<0.01). p53 protein was not detected by immunohistochemistry in all groups before the 75^th wk of the experiment. At the 105^th wk, the positive rates fo p53 were 78.6%, 60% and 71.4% in groups A, B and C, respectively, which were significantly higher than that in group D (10%) (all P<0.05). An abnormal band of p53 gene was observed in groups A and C. The mutation points of p53gene in tree shrews with HCC were at codons 275, 78 and 13. The nucleotide sequence and amino acid sequence of tree shrew's wild-type p53 showed 91.7% and 93.4% homologies with those of human p53,respectively. The immunopositivity for p21 was found before HCC development. The incidence of HCC was significantly higher in tree shrews that were positive for p21 than those negative for p21 (80.0% vs 11.0%, P<0.001).The incidence of HCC in p21 positive animals in group A was significantly higher than those positive for p21 in group C (P<O.05).CONCLUSION: A remarkable synergistic effect on HCC development exists between HBV and AFB1. p53 mutation promotes the development of HCC. HBV and AFB1 may synergistically induce p53 gene mutation, and stimulate ras gene expression, ras gene is activated at the earlier stage during hepatocarcinogenesis, p21 protein may be an early marker, and the alterations of p53 may be a late event in the development of HCC.展开更多
Previous studies have demonstrated the possible role of several aquaporins in seed germination. But systematic investigation of the role ofaquaporin family members in this process is lacking. Here, the developmental r...Previous studies have demonstrated the possible role of several aquaporins in seed germination. But systematic investigation of the role ofaquaporin family members in this process is lacking. Here, the developmental regulation of plasma membrane intrinsic protein （PIP） expression throughout germination and post-germination processes in rice embryos was analyzed. The expression patterns of the PIPs suggest these aquaporins play different roles in seed germination and seedling growth. Partial silencing of the water channel genes, OsPIP1;1 and OsPIP1：3, reduced seed germination while over-expression of OsPIP1：3 promoted seed germination under water-stress conditions. Moreover, spatial expression analysis indicates that OsPIP1：3 is expressed predominantly in embryo during seed germination. Our data also revealed that the nitric oxide （NO） donors, sodium nitroprusside （SNP） and S-nitrosoglutathione （GSNO）, promoted seed germination; furthermore, the NO scavenger, 2-（4-carboxyphenyl）-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide, inhibited germination and reduced the stimulative effects of SNP and GSNO on rice germination. Exogenous NO stimulated the transcription of OsPIP1：1, OsPIP1：2, OsPIP1：3 and OsPIP2：8 in germinating seeds. These results suggest that water channels play an important role in seed germination, acting, at least partly, in response to the NO signaling pathway.展开更多
Gastric cancer is a complex disease that is affected by multiple genetic and environmental factors. For the precise diagnosis and effective treatment of gastric cancer, the heterogeneity of the disease must be simplif...Gastric cancer is a complex disease that is affected by multiple genetic and environmental factors. For the precise diagnosis and effective treatment of gastric cancer, the heterogeneity of the disease must be simplified; one way to achieve this is by dividing the disease into subgroups. Toward this effort, recent advances in high-throughput sequencing technology have revealed four molecular subtypes of gastric cancer, which are classified as Epstein-Barr viruspositive, microsatellite instability, genomically stable, and chromosomal instability subtypes. We anticipate that this molecular subtyping will help to extend our knowledge for basic research purposes and will be valuable for clinical use. Here, we review the genomic and epigenomic heterogeneity of the four molecular subtypes of gastric cancer. We also describe a mutational meta-analysis and a reanalysis of DNA methylation that were performed using previously reported gastric cancer datasets.展开更多
BACKGROUND Genomic profiling of tumors has contributed to the understanding of colorectal cancer(CRC), facilitating diagnosis, prognosis and selection of treatments,including targeted regimens. A report suggested that...BACKGROUND Genomic profiling of tumors has contributed to the understanding of colorectal cancer(CRC), facilitating diagnosis, prognosis and selection of treatments,including targeted regimens. A report suggested that a 19-gene-based risk classifier(TCA19) was a prognostic tool for patients with stage III CRC. The survival outcomes in patients with stage IV CRC are still poor and appropriate selection of targeted therapies and immunotherapies is challenging.AIM To assess clinical implication of TCA19 in patients with stage IV CRC, and to identify TCA19 with involvement in immune-oncology.METHODS A retrospective review of the medical records of 60 patients with stage IV CRC was conducted, assessing clinicopathological variables and progression-free survival(PFS). TCA19 gene expression was determined by quantitative polymerase chain reaction(qPCR) in matched normal and tumor tissues taken from the study cohort. Expression of potential immune-oncology regulatory proteins and targets was examined by immunohistochemistry(IHC), western blot, immunofluorescence staining in tissues from a validation cohort of 10 patients, and in CRC cell lines co-cultured with monocyte in vitro.RESULTS In the patients with TCA19 score higher than the median, the PFS rates of eight patients who received the targeted regimens were significantly higher than the PFS rates of four patients who received 5-fluorouracil-based regimen(P = 0.041).In multivariate analysis, expression of signaling lymphocytic activation molecule family, member 7(SLAMF7) and triggering receptor expressed on myeloid cells 1(TREM1) was associated with PFS in the 60-patient cohort. After checking another 10 validate set, the expression of the IHC, the level of real-time qPCR,and the level of western blot were lower for SLAMF7 and higher for TREM7 in primary and metastatic tumors than in normal tissues. In CRC cells expressing SLAMF7 that were co-cultured with a monocytic cell line, levels of CD 68 and CD73 were significantly lower at day 5 of co-culture than at day 0.CONCLUSION The TCA19 score might be prognostic for target-regimen-specific PFS in stage IV CRC. Down-regulation of SLAMF7 and up-regulation of TREM1 occur in primary and metastatic tumor tissues.展开更多
MicroRNAs（miRNAs） are a class of highly conserved small non-coding RNA molecules that play a pivotal role in several cellular functions.In this study,miRNA and messenger RNA（mRNA） profiles were examined by Illumin...MicroRNAs（miRNAs） are a class of highly conserved small non-coding RNA molecules that play a pivotal role in several cellular functions.In this study,miRNA and messenger RNA（mRNA） profiles were examined by Illumina microarray in mouse embryonic stem cells（ESCs） derived from parthenogenetic,androgenetic,and fertilized blastocysts.The global analysis of miRNA-mRNA target pairs provided insight into the role of miRNAs in gene expression.Results showed that a total of 125 miRNAs and 2394 mRNAs were differentially expressed between androgenetic ESCs（aESCs） and fertilized ESCs（fESCs）,a total of 42 miRNAs and 87 mRNAs were differentially expressed between parthenogenetic ESCs（pESCs） and fESCs,and a total of 99 miRNAs and 1788 mRNAs were differentially expressed between aESCs and pESCs.In addition,a total of 575,5 and 376 miRNA-mRNA target pairs were observed in aESCs vs.fESCs,pESCs vs.fESCs,and aESCs vs.pESCs,respectively.Furthermore,15 known imprinted genes and 16 putative uniparentally expressed miRNAs with high expression levels were confirmed by both microarray and real-time RT-PCR.Finally,transfection of miRNA inhibitors was performed to validate the regulatory relationship between putative maternally expressed miRNAs and target mRNAs. Inhibition of miR-880 increased the expression of Peg3,Dyrklb,and Prrg2 mRNA,inhibition of miR-363 increased the expression of Nfat5 and Soatl mRNA,and inhibition of miR-883b-5p increased Nfat5,Tacstd2,and Ppapdc1 mRNA.These results warrant a functional study to fully understand the underlying regulation of genomic imprinting in early embryo development.展开更多
Infectious agents causing aborted fetus problems in domestic pigs were investigated in this study. More than 10 different infectious agents were known to cause abortion in swine and the major eight viruses among them ...Infectious agents causing aborted fetus problems in domestic pigs were investigated in this study. More than 10 different infectious agents were known to cause abortion in swine and the major eight viruses among them were inspected. One hundred twelve samples of aborted fetuses from nine provinces in South Korea were collected during April to November, 2013 in this study for the diagnosis of infectious agents causing abortions in pigs. Eight major infection viruses were examined in this study mainly using various diagnostic kits and reverse transcriptase polymerase chain reaction （RT-PCR）. Positive rate of the detection differed from each viruses. In this study, the main focus was the porcine reproductive and respiratory syndrome virus （PRRSV）, which took the second large portion in the positive rate of detection, and then its ORF5 gene was compared with modified live virus （MLV） vaccine strain to figure out the influence of vaccine on disease. Between four positive samples＇ sequence, two of them were 99.9%-100% similar to MLV vaccine strain and two other samples were 88.6%-92.7% similar. Similarity rate of the sequences between the vaccine and virus from aborted fetuses are very crucial, because it implies that abortion in swine can be made due to the usage of vaccine not only by the infection of field virus, and if MLV vaccine actually do have an impact on the infection, usage of the vaccine should be reconsidered.展开更多
CD （Cryptocarya densiflora） Blume has traditionally been used as an herbal medicine. In this study, the effects of CDEE （CD ethanol extract） on inflammation were investigated in LPS （lipopolysaccharide）-stimulat...CD （Cryptocarya densiflora） Blume has traditionally been used as an herbal medicine. In this study, the effects of CDEE （CD ethanol extract） on inflammation were investigated in LPS （lipopolysaccharide）-stimulated mouse RAW264.7 macrophages. We investigated the effects of CDEE on the production of NO, PGE2 interleukin （IL）-6, IL-1β, and tumor necrosis factor （TNF）-α in LPS-stimulated RAW264.7 cells. We measured the mRNA or protein expression of the pro-inflammatory mediators induced by CDEE in LPS-stimulated RAW264.7 cells. We explored the expression of Nrf-2, heme oxygenase （HO）-I and NADPH-quinone oxidoreductase （NQO）-I to elucidate the antioxidative mechanisms. CDEE significantly inhibited the production of NO, PGE2, IL-6, IL-1β and TNF-α in LPS-stimulated RAW264.7 cells. CDEE suppressed the mRNA or protein expression of iNOS, COX-2, and the MAPKs with a reduction in the translocation of NF-κB in LPS-stimulated RAW264.7 cells. In addition, CDEE significantly increased the expression of HO-I and NQO-1 with an increase in the translocation of Nrf-2 into the nucleus. These results indicate that CDEE inhibits the LPS-induced inflammatory and oxidative responses via suppression of NF-κB activation and the enhancement of Nrf2 activation. We suggest that CDEE may be therapeutic for treating inflammatory diseases.展开更多
Influenza,a highly contagious respiratory infectious disease caused by an influenza virus,is a threat to public health worldwide.Avian influenza viruses(AIVs)have the potential to cause the next pandemic by crossing t...Influenza,a highly contagious respiratory infectious disease caused by an influenza virus,is a threat to public health worldwide.Avian influenza viruses(AIVs)have the potential to cause the next pandemic by crossing the species barrier through mutation of viral genome.Here,we investigated the pathogenicity of AIVs obtained from South Korea and Mongolia during 2018–2019 by measuring viral titers in the lungs and extrapulmonary organs of mouse models.In addition,we assessed the pathogenicity of AIVs in ferret models.Moreover,we compared the ability of viruses to replicate in mammalian cells,as well as the receptor-binding preferences of AIV isolates.Genetic analyses were finally performed to identify the genetic relationships and amino acid substitutions between viral proteins during mammalian adaptation.Of the 24 AIV isolates tested,A/Mallard/South Korea/KNU2019-34/2019(KNU19-34;H1N1)caused severe bodyweight loss and high mortality in mice.The virus replicated in the lungs,kidneys,and heart.Importantly,KNU19-34-infected ferrets showed high viral loads in both nasal washes and lungs.KNU19-34 replicated rapidly in A549 and bound preferentially to human likeα2,6-linked sialic acids rather than to avian-likeα2,3-linked sialic acids,similar to the pandemic A/California/04/2009(H1N1)strain.Gene segments of KNU19-34 were distributed in Egypt and Asia lineages from 2015 to 2018,and the virus had several amino acid substitutions compared to H1N1 AIV isolates that were non-pathogenic in mice.Collectively,the data suggest that KNU19-34 has zoonotic potential and the possibility of new mutations responsible for mammalian adaptation.展开更多
The bioconversion of 4-hydroxy-2-keto acid derivatives via aldol condensation of formaldehyde and pyruvate has received substantial attention as potential source of chemicals for production of amino acids,hydroxy carb...The bioconversion of 4-hydroxy-2-keto acid derivatives via aldol condensation of formaldehyde and pyruvate has received substantial attention as potential source of chemicals for production of amino acids,hydroxy carboxylic acids,and chiral aldehydes.We developed an environmentally friendly biocatalyst consisting of a novel thermostable class II pyruvate aldolase from Deinococcus radiodurans with maltose-binding protein(MBP-DrADL),which has specific activity of 46.3μmol min-1 mg-1.Surprisingly,MBP-DrADL maintained over 60%of enzyme activity for 4 days at 50 to 65°C,we used MBP-DrADL as the best candidate enzyme to produce 2-keto-4-hydroxybutyrate(2-KHB)from formaldehyde and pyruvate via aldol condensation.The optimum reaction conditions for 2-KHB production were 50°C,pH 8.0,5 mM Mg2+,100 mM formaldehyde,and 200 mM pyruvate.Under these optimized conditions,MBP-DrADL produced 76.5 mM(8.94 g L-1)2-KHB over 60 min with a volumetric productivity of 8.94 g L-1 h-1 and a specific productivity of 357.6 mg mg-enzyme-1 h-1.Furthermore,2-KHB production was improved by continuous addition of substrates,which produced approximately 124.8 mM(14.6 g L-1)of 2-KHB over 60 min with a volumetric productivity and specific productivity of 14.6 g L-1 h-1 and 583.4 mg mg-enzyme-1 h-1,respectively.MBP-DrADL showed the highest specific productivity for 2-KHB production yet reported.Our study provides a highly efficient biocatalyst for the synthesis of 2-KHB and lays the foundation for large-scale production and application of high-value compounds from formaldehyde.展开更多
Background:Picropodophllotoxin(PPT),a principal component of Podophyllum hexandrum root,demonstrates various beneficial biological activities in multiple cancer types,including antitumor and antiproliferative properti...Background:Picropodophllotoxin(PPT),a principal component of Podophyllum hexandrum root,demonstrates various beneficial biological activities in multiple cancer types,including antitumor and antiproliferative properties.Despite its known effects,the specific mechanisms by which PPT induces apoptosis in oral squamous cell carcinoma(OSCC)cells lack full clarification.Aims:This study aimed to evaluate the role of PPT in inducing apoptosis in OSCC cells by targeting signal transducer and activator of transcription 3(STAT3)and to investigate the underlying molecular pathways.Methods:Human OSCC cell lines(HN22 and HSC4)were treated with PPT.Cell viability,colony formation,and apoptotic morphological changes were evaluated.Reactive oxygen species(ROS)generation and mitochondrial function were assessed using tetramethyl rhodamine methyl ester,MitoSOX,and 2′,7′-dichlorodihydrofluorescein diacetate(DCFH-DA)assays following PPT treatment.The expression of apoptosis markers,including cleaved Poly(ADP-Ribose)Polymerase(c-PARP)and other target proteins,was measured using western blotting.ROS involvement was further confirmed using the ROS scavenger N-acetylcysteine(NAC).Results:Treatment with PPT resulted in a substantial reduction in cell viability,a decrease in colony formation capacity,and evident morphological changes in OSCC cells.These effects were dose-and time-dependent,as evidenced by increased expression of c-PARP.PPT-induced apoptosis was mediated by excessive ROS generation,which was almost completely blocked by NAC pretreatment.Conclusions:These findings suggest that PPT may serve as a promising therapeutic agent for treating human oral cancer by inhibiting the STAT3 pathway and inducing ROS-mediated apoptosis.展开更多
Objectives:Limosilactobacillus reuteri is a beneficial Lactobacillus widely used in foods and supplements to promote overall health.Some studies also suggest it supports skin health and prevents allergies and cardiova...Objectives:Limosilactobacillus reuteri is a beneficial Lactobacillus widely used in foods and supplements to promote overall health.Some studies also suggest it supports skin health and prevents allergies and cardiovascular disease.However,research on its skin-protective effects against photoaging has not been conducted.This study evaluated the potential of culture broths fromthree L.reuteri strains(DS0333,DS0384,and DS0385)to inhibit skin photoaging.Methods:To assess their anti-photoaging potential,the culture broths were examined for antioxidant capacity,melanin inhibition,and collagen synthesis promotion.Radical scavenging activity was tested using 2,2-diphenyl-1-picrylhydrazyl(DPPH)and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS)assays.The biosynthetic activity of melanin and associated protein markers involved in melanogenesis was examined in a B16F10 mouse melanoma model.Type I procollagen synthesis and matrix metalloproteinase-1(MMP-1)inhibition were evaluated in ultraviolet B(UVB)-damaged human dermal fibroblasts(HDFs).Results:The culture broths exhibited concentrationdependent antioxidant activity and significantly suppressed melanin synthesis triggered byα-melanocyte-stimulating hormone(α-MSH).Transcription factors involved inmelanogenesis,namelymicrophthalmia-associated transcription factor(MITF),tyrosinase-related protein 1(TRP-1),and 2(TRP-2),were significantly downregulated following treatment.Treatment with culture broths also enhanced type I procollagen production and inhibited MMP-1 activity and protein expression in UVB-exposed HDFs.Among the strains,DS0333 demonstrated the strongest efficacy and was further investigated.It enhanced the proliferation of skin cells and attenuated the levels of age-associated markers such as MMP-1,mitogen-activated protein kinase(MAPK),and activator protein 1(AP-1).High-performance liquid chromatography(HPLC)analysis identified phenyllactic acid(PLA)as the predominant active compound.Conclusions:These results indicate that DS0333 culture broth exhibits strong anti-aging effects and can be applied in functional cosmetics aimed at promoting skin health.展开更多
Infiltration and activation of peripheral immune cells are critical in the progression of multiple sclerosis and its experimental animal model,experimental autoimmune encephalomyelitis(EAE).This study investigates the...Infiltration and activation of peripheral immune cells are critical in the progression of multiple sclerosis and its experimental animal model,experimental autoimmune encephalomyelitis(EAE).This study investigates the role of high mobility group box 1(HMGB1)in oligodendrocyte precursor cells(OPCs)in modulating pathogenic T cells infiltrating the central nervous system through the blood-brain barrier(BBB)by using OPC-specific HMGB1 knockout(KO)mice.We found that HMGB1 released from OPCs promotes BBB disruption,subsequently allowing increased immune cell infiltration.The migration of CD4+T cells isolated from EAE-induced mice was enhanced when co-cultured with OPCs compared to oligodendrocytes(OLs).OPC-specific HMGB1 KO mice exhibited lower BBB permeability and reduced immune cell infiltration into the CNS,leading to less damage to the myelin sheath and mitigated EAE progression.CD4+T cell migration was also reduced when co-cultured with HMGB1 knock-out OPCs.Our findings reveal that HMGB1 secretion from OPCs is crucial for regulating immune cell infiltration and provides insights into the immunomodulatory function of OPCs in autoimmune diseases.展开更多
Long term hepatitis B virus (HBV) infection is a major risk factor in pathogenesis of chronic liver diseases,including hepatocellular carcinoma (HCC). The HBV encod-ed proteins,hepatitis B virus X protein and preS,app...Long term hepatitis B virus (HBV) infection is a major risk factor in pathogenesis of chronic liver diseases,including hepatocellular carcinoma (HCC). The HBV encod-ed proteins,hepatitis B virus X protein and preS,appear to contribute importantly to the pathogenesis of HCC. Both are associated with oxidative stress,which can damage cellular molecules like lipids,proteins,and DNA during chronic infection. Chronic alcohol use is another important factor that contributes to oxidative stress in the liver. Previous studies reported that treatment with antioxidants,such as curcumin,silymarin,green tea,and vitamins C and E,can protect DNA from damage and regulate liver pathogenesis-related cascades by reducing reactive oxygen species. This review summarizes some of the relationships between oxidative stress and liver pathogenesis,focusing upon HBV and alcohol,and suggests antioxidant therapeutic approaches.展开更多
AIM:To investigate the role of hepatitis B virus X-protein(HBx)-induced reactive oxygen species(ROS)on liver carcinogenesis in HBx transgenic mice and HepG2-HBx cells.METHODS:Cell growth rate was analyzed,and through ...AIM:To investigate the role of hepatitis B virus X-protein(HBx)-induced reactive oxygen species(ROS)on liver carcinogenesis in HBx transgenic mice and HepG2-HBx cells.METHODS:Cell growth rate was analyzed,and through western blotting,mitogenic signaling was observed.Endogenous ROS from wild and HBx transgenic mice and HepG2-Mock and HBx cells were assayed by FACS-calibur.Identification of oxidized and reduced phosphatase and tensin homolog(PTEN)was analyzed through N-ethylmaleimide alkylation,nonreducing electrophoresis.RESULTS:We observed that the cell-proliferation-related phosphoinositide 3-kinase/Akt pathway is activated by HBx in vivo and in vitro.Increased ROS were detected by HBx.Tumor suppressor PTEN,via dephosphorylation of Akt,was oxidized and inactivated by increased ROS.Increased oxidized PTEN activated the mitogenic pathway through over-activated Akt.However,treatment with ROS scavenger N-acetyl cysteine can reverse PTEN to a reduced form.Endogenously produced ROS also stimulated HBx expression.CONCLUSION:HBx induced ROS promoted Akt pathways via oxidized inactive PTEN.HBx and ROS maintained a positive regulatory loop,which aggravated carcinogenesis.展开更多
Soil salinity causes the negative effects on the growth and yield of crops. In this study, two sweet potato （Ipomoea batatas L.） cultivars, Xushu 28 （X-28） and Okinawa 100 （O-100）, were examined under 50 and 100...Soil salinity causes the negative effects on the growth and yield of crops. In this study, two sweet potato （Ipomoea batatas L.） cultivars, Xushu 28 （X-28） and Okinawa 100 （O-100）, were examined under 50 and 100 mmol L-1 NaCI stress. X-28 cultivar is relatively high salt tolerant than O-100 cultivar. Interestingly, real-time quantitative polymerase chain reaction （RT-qPCR） results indicated that sweet potato high-affinity K^＋ transporter 1 （IbHKT1） gene expression was highly induced by 50 and 100 mmol L-1 NaCI stress in the stems of X-28 cultivar than in those of O-100 cultivar, but only slightly induced by these stresses in the leaves and fibrous roots in both cultivars. To characterize the function of IbHKT1 transporter, we performed ion-flux analysis in tobacco transient system and yeast complementation. Tobacco transient assay showed that IbHKT1 could uptake sodium （Na^＋）. Yeast complementation assay showed that IbHKT1 could take up K^＋ in 50 mmol L^-1 K^＋ medium without the presence of NaCI. Moreover, Na^＋ uptake significantly increased in yeast overexpressing IbHKTI. These results showed that IbHKT1 transporter could have K^＋-Na^＋ symport function in yeast. Therefore, the modes of action of IbHKT1 in transgenic yeast could differ from the mode of action of the other HKT1 transporters in class I. Potentially, IbHKT1 could be used to improve the salt tolerance nature in sweet potato.展开更多
Chikungunya fever is a vector-borne viral disease transmitted to humans by chikungunya virus(CHIKV)-infected mosquitoes.There have been many outbreaks of CHIKV infection worldwide,and the virus poses ongoing risks to ...Chikungunya fever is a vector-borne viral disease transmitted to humans by chikungunya virus(CHIKV)-infected mosquitoes.There have been many outbreaks of CHIKV infection worldwide,and the virus poses ongoing risks to global health.To prevent and control CHIKV infection,it is important to improve the current CHIKV diagnostic approaches to allow for the detection of low CHIKV concentrations and to correctly distinguish CHIKV infections from those due to other mosquito-transmitted viruses,including dengue virus(DENV),Japanese encephalitis virus(JEV),and Zika virus(ZIKV).Here,we produced monoclonal antibodies(mAbs)against the CHIKV envelope 2 protein(CHIKV-E2)and compared their sensitivity and specificity with commercially available mAbs using enzyme-linked immunosorbent assays(ELISA).Two anti-CHIKV-E2 mAbs,19-1 and 21-1,showed higher binding affinities to CHIKV-E2 protein than the commercial mAbs did.In particular,the 19-1 mAb had the strongest binding affinity to inactivated CHIKV.Moreover,the 19-1 mAb had very little cross-reactivity with other mosquito-borne viruses,such as ZIKV,JEV,and DENV.These results suggest that the newly produced anti-CHIKV-E2 mAb,19-1,could he used for CHIKV diagnostic approaches.展开更多
Clusterin, a protein associated with multiple functions, is expressed in a wide variety of mammalian tissues. Although clusterin is known to be involved in neurodegenerative diseases, ageing, and tumorigenesis, a deta...Clusterin, a protein associated with multiple functions, is expressed in a wide variety of mammalian tissues. Although clusterin is known to be involved in neurodegenerative diseases, ageing, and tumorigenesis, a detailed analysis of the consequences of gain- or loss-of- function approaches has yet to be performed to understand the underlying mechanisms of clusterin functions. Since clusterin levels change in neurological diseases, it is likely that clusterin contributes to cell death and degeneration in general. Zebrafish was investigated as a model system to study human diseases. During development, zebrafish clusterin was expressed in the notochord and nervous system. Embryonic overexpression of clusterin by mRNA microinjection did not affect axis formation, whereas its knock-down by anti-sense morpholino treatment resulted in neuronal cell death. To analyze the function of clusterin in neurodegeneration, a transgenic zebrafish was investigated, in which nitroreductase expression is regulated under the control of a neuron-specifc huC promoter which is active between the stages of early neuronal precursors and mature neurons. Nitroreductase turns metronidazole into a cytotoxic agent that induces cell death within 12 h. After metronidazole treatment, transgenic zebrafish showed neuron-specific cell death. Interestingly, we also observed a dramatic induction of clusterin expression in the brain and spinal cord in these fish, suggesting a direct or indirect role of clusterin in neuronal cell death and thus, more generally, in neurodegeneration.展开更多
Background Anethole(AN)is an organic antioxidant compound with a benzene ring and is expected to have a positive impact on early embryogenesis in mammals.However,no study has examined the effect of AN on porcine embry...Background Anethole(AN)is an organic antioxidant compound with a benzene ring and is expected to have a positive impact on early embryogenesis in mammals.However,no study has examined the effect of AN on porcine embryonic development.Therefore,we investigated the effect of AN on the development of porcine embryos and the underlying mechanism.Results We cultured porcine in vitro-fertilized embryos in medium with AN(0,0.3,0.5,and 1 mg/mL)for 6 d.AN at 0.5 mg/mL significantly increased the blastocyst formation rate,trophectoderm cell number,and cellular survival rate compared to the control.AN-supplemented embryos exhibited significantly lower reactive oxygen species levels and higher glutathione levels than the control.Moreover,AN significantly improved the quantity of mitochondria and mitochondrial membrane potential,and increased the lipid droplet,fatty acid,and ATP levels.Interestingly,the levels of proteins and genes related to the sonic hedgehog(SHH)signaling pathway were significantly increased by AN.Conclusions These results revealed that AN improved the developmental competence of porcine preimplantation embryos by activating SHH signaling against oxidative stress and could be used for large-scale production of high-quality porcine embryos.展开更多
Experimental stroke research commonly employs focal cerebral ischemic rat models （Bederson et al., 1986a; Longa et al., 1989）. In human patients, ischemic stroke typically results from thrombotic or embolic occlusio...Experimental stroke research commonly employs focal cerebral ischemic rat models （Bederson et al., 1986a; Longa et al., 1989）. In human patients, ischemic stroke typically results from thrombotic or embolic occlusion of a major cerebral artery, usually the mid- dle cerebral artery （MCA）. Experimental focal cerebral ischemia models have been employed to mimic human stroke （Durukan and Tatlisumak, 2007）. Rodent models of focal cerebral ischemia that do not require craniotomy have been developed using intraluminal suture occlusion of the MCA （MCA occlusion, MCAO） （Rosamond et al., 2008）. Furthermore, mouse MCAO models have been wide- ly used and extended to genetic studies of cell death or recovery mechanisms （Liu and McCullough, 2011）. Genetically engineered mouse stroke models are particularly useful for evaluation of isch- emic pathophysiology and the design of new prophylactic, neuro- protective, and therapeutic agents and interventions （Armstead et al., 2010）. During the past two decades, MCAO surgical techniques have been developed that do not reveal surgical techniques for mouse MCAO model engineering. Therefore, we compared MCAO surgical methods in rats and mice.展开更多
AIM:To investigate the effect of methylsulfonylmethane(MSM), recently reported to have anti-cancer effects, in liver cancer cells and transgenic mice. METHODS: Three liver cancer cell lines, HepG2, Huh7-Mock and Huh7-...AIM:To investigate the effect of methylsulfonylmethane(MSM), recently reported to have anti-cancer effects, in liver cancer cells and transgenic mice. METHODS: Three liver cancer cell lines, HepG2, Huh7-Mock and Huh7-H-rasG12V, were used. Cell growth was measured by Cell Counting Kit-8 and soft agar assay. Western blot analysis was used to detect caspases, poly(ADP-ribose) polymerase(PARP), and B-cell lymphoma2(Bcl-2) expressions. For in vivo study, we administered MSM to H-ras12V transgenic mice for 3 mo. RESULTS: MSM decreased the growth of HepG2, Huh7-Mock and Huh7-H-rasG12V cells in a dose-dependent manner. That was correlated with significantly increased apoptosis and reduced cell numbers in MSM treated cells. Cleaved caspase-8, cleaved caspase-3 and cleaved PARP were remarkably increased in the liver cancer cells treated with 500 mmol/L of MSM; however, Bcl-2 was slightly decreased in 500 mmol/L. Liver tumor development was greatly inhibited in the H-ras12V transgenic mice treated with MSM, compared to control, by showing reduced tumor size and number. Cleaved PARP was significantly increased in non-tumor treated with MSM compared to control. CONCLUSION: Liver injury was also significantly attenuated in the mice treated with MSM. Taken together, all the results suggest that MSM has anti-cancer effects through inducing apoptosis in liver cancer.展开更多
基金Supported by the National Natural Science Foundation of China,No.39260033Natural Science Foundation of Guangxi,No.0143058
文摘AIM: To investigate p53 mutation and p21 expression in hepatocarcinogenesis induced by hepatitis B virus (HBV) and aflatoxin B1 (AFB1) in tree shrews, and to reveal the role of these genes in hepatocarcinogenesis.METHODS: Tree shrews were divided into four groups:group A, those infected with HBV and fed with AFB1 (n = 39);group B, those infected with HBV alone (n = 28); group C,those fed with AFB1 alone (n = 29); and group D, normal controls (n = 20). The tree shrews underwent liver biopsies once every 15 wk. Expression of p53 and p21 proteins and genes in the biopsies and tumor tissues of the experimental tree shrews was detected, respectively, by immunohistochemistry,and by Southem blotting and reverse transcription-polymerase chain reaction and sequencing.RESULTS: The incidence of hepatocellular carcinomas (HCC) was higher in group A (66.7%) than that in group B (3.57%) and C (30%). The time of HCC occurrence was also earlier in group A than that in group C (120.0±16.6 wk vs 153.3±5.8 wk, respectively, P<0.01). p53 protein was not detected by immunohistochemistry in all groups before the 75^th wk of the experiment. At the 105^th wk, the positive rates fo p53 were 78.6%, 60% and 71.4% in groups A, B and C, respectively, which were significantly higher than that in group D (10%) (all P<0.05). An abnormal band of p53 gene was observed in groups A and C. The mutation points of p53gene in tree shrews with HCC were at codons 275, 78 and 13. The nucleotide sequence and amino acid sequence of tree shrew's wild-type p53 showed 91.7% and 93.4% homologies with those of human p53,respectively. The immunopositivity for p21 was found before HCC development. The incidence of HCC was significantly higher in tree shrews that were positive for p21 than those negative for p21 (80.0% vs 11.0%, P<0.001).The incidence of HCC in p21 positive animals in group A was significantly higher than those positive for p21 in group C (P<O.05).CONCLUSION: A remarkable synergistic effect on HCC development exists between HBV and AFB1. p53 mutation promotes the development of HCC. HBV and AFB1 may synergistically induce p53 gene mutation, and stimulate ras gene expression, ras gene is activated at the earlier stage during hepatocarcinogenesis, p21 protein may be an early marker, and the alterations of p53 may be a late event in the development of HCC.
基金This work was supported by the National Natural Science Foundation of China(No.30670172)by the Korea Foundation for International Cooperation of Science and Technology(K/C0S)through a grant provided by the Korean Ministry of Science and Technology.
文摘Previous studies have demonstrated the possible role of several aquaporins in seed germination. But systematic investigation of the role ofaquaporin family members in this process is lacking. Here, the developmental regulation of plasma membrane intrinsic protein （PIP） expression throughout germination and post-germination processes in rice embryos was analyzed. The expression patterns of the PIPs suggest these aquaporins play different roles in seed germination and seedling growth. Partial silencing of the water channel genes, OsPIP1;1 and OsPIP1：3, reduced seed germination while over-expression of OsPIP1：3 promoted seed germination under water-stress conditions. Moreover, spatial expression analysis indicates that OsPIP1：3 is expressed predominantly in embryo during seed germination. Our data also revealed that the nitric oxide （NO） donors, sodium nitroprusside （SNP） and S-nitrosoglutathione （GSNO）, promoted seed germination; furthermore, the NO scavenger, 2-（4-carboxyphenyl）-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide, inhibited germination and reduced the stimulative effects of SNP and GSNO on rice germination. Exogenous NO stimulated the transcription of OsPIP1：1, OsPIP1：2, OsPIP1：3 and OsPIP2：8 in germinating seeds. These results suggest that water channels play an important role in seed germination, acting, at least partly, in response to the NO signaling pathway.
基金Supported by Grants from the genomics program of the National Research Foundation of Korea funded by the Ministry of ScienceICT+4 种基金and Future PlanningNRF-2012M3A9D1054670 and NRF-2014M3C9A3068554(to Kim SY)Basic Science Research Program through the National Research Foundation of Korea funded by the Ministry of EducationNRF-2013R1A1A2006621(to Kim M)the Korea Research Institute of Bioscience and Biotechnology research initiative grant
文摘Gastric cancer is a complex disease that is affected by multiple genetic and environmental factors. For the precise diagnosis and effective treatment of gastric cancer, the heterogeneity of the disease must be simplified; one way to achieve this is by dividing the disease into subgroups. Toward this effort, recent advances in high-throughput sequencing technology have revealed four molecular subtypes of gastric cancer, which are classified as Epstein-Barr viruspositive, microsatellite instability, genomically stable, and chromosomal instability subtypes. We anticipate that this molecular subtyping will help to extend our knowledge for basic research purposes and will be valuable for clinical use. Here, we review the genomic and epigenomic heterogeneity of the four molecular subtypes of gastric cancer. We also describe a mutational meta-analysis and a reanalysis of DNA methylation that were performed using previously reported gastric cancer datasets.
基金Korea Research Foundation,No.2016R1E1A1A02919844 to Kim JC and No.2017R1A2B1009062 to Roh SAMinistry of Science,ICT,and Future Planning,Republic of Koreathe Asan Institute for Life Sciences,No.2016-710 to Lee JL
文摘BACKGROUND Genomic profiling of tumors has contributed to the understanding of colorectal cancer(CRC), facilitating diagnosis, prognosis and selection of treatments,including targeted regimens. A report suggested that a 19-gene-based risk classifier(TCA19) was a prognostic tool for patients with stage III CRC. The survival outcomes in patients with stage IV CRC are still poor and appropriate selection of targeted therapies and immunotherapies is challenging.AIM To assess clinical implication of TCA19 in patients with stage IV CRC, and to identify TCA19 with involvement in immune-oncology.METHODS A retrospective review of the medical records of 60 patients with stage IV CRC was conducted, assessing clinicopathological variables and progression-free survival(PFS). TCA19 gene expression was determined by quantitative polymerase chain reaction(qPCR) in matched normal and tumor tissues taken from the study cohort. Expression of potential immune-oncology regulatory proteins and targets was examined by immunohistochemistry(IHC), western blot, immunofluorescence staining in tissues from a validation cohort of 10 patients, and in CRC cell lines co-cultured with monocyte in vitro.RESULTS In the patients with TCA19 score higher than the median, the PFS rates of eight patients who received the targeted regimens were significantly higher than the PFS rates of four patients who received 5-fluorouracil-based regimen(P = 0.041).In multivariate analysis, expression of signaling lymphocytic activation molecule family, member 7(SLAMF7) and triggering receptor expressed on myeloid cells 1(TREM1) was associated with PFS in the 60-patient cohort. After checking another 10 validate set, the expression of the IHC, the level of real-time qPCR,and the level of western blot were lower for SLAMF7 and higher for TREM7 in primary and metastatic tumors than in normal tissues. In CRC cells expressing SLAMF7 that were co-cultured with a monocytic cell line, levels of CD 68 and CD73 were significantly lower at day 5 of co-culture than at day 0.CONCLUSION The TCA19 score might be prognostic for target-regimen-specific PFS in stage IV CRC. Down-regulation of SLAMF7 and up-regulation of TREM1 occur in primary and metastatic tumor tissues.
基金supported by the grants from the NextGeneration BioGreen 21 Program(Nos.PJ009080 and PJ00909801)Rural Development Administration,Republic of Korea
文摘MicroRNAs（miRNAs） are a class of highly conserved small non-coding RNA molecules that play a pivotal role in several cellular functions.In this study,miRNA and messenger RNA（mRNA） profiles were examined by Illumina microarray in mouse embryonic stem cells（ESCs） derived from parthenogenetic,androgenetic,and fertilized blastocysts.The global analysis of miRNA-mRNA target pairs provided insight into the role of miRNAs in gene expression.Results showed that a total of 125 miRNAs and 2394 mRNAs were differentially expressed between androgenetic ESCs（aESCs） and fertilized ESCs（fESCs）,a total of 42 miRNAs and 87 mRNAs were differentially expressed between parthenogenetic ESCs（pESCs） and fESCs,and a total of 99 miRNAs and 1788 mRNAs were differentially expressed between aESCs and pESCs.In addition,a total of 575,5 and 376 miRNA-mRNA target pairs were observed in aESCs vs.fESCs,pESCs vs.fESCs,and aESCs vs.pESCs,respectively.Furthermore,15 known imprinted genes and 16 putative uniparentally expressed miRNAs with high expression levels were confirmed by both microarray and real-time RT-PCR.Finally,transfection of miRNA inhibitors was performed to validate the regulatory relationship between putative maternally expressed miRNAs and target mRNAs. Inhibition of miR-880 increased the expression of Peg3,Dyrklb,and Prrg2 mRNA,inhibition of miR-363 increased the expression of Nfat5 and Soatl mRNA,and inhibition of miR-883b-5p increased Nfat5,Tacstd2,and Ppapdc1 mRNA.These results warrant a functional study to fully understand the underlying regulation of genomic imprinting in early embryo development.
文摘Infectious agents causing aborted fetus problems in domestic pigs were investigated in this study. More than 10 different infectious agents were known to cause abortion in swine and the major eight viruses among them were inspected. One hundred twelve samples of aborted fetuses from nine provinces in South Korea were collected during April to November, 2013 in this study for the diagnosis of infectious agents causing abortions in pigs. Eight major infection viruses were examined in this study mainly using various diagnostic kits and reverse transcriptase polymerase chain reaction （RT-PCR）. Positive rate of the detection differed from each viruses. In this study, the main focus was the porcine reproductive and respiratory syndrome virus （PRRSV）, which took the second large portion in the positive rate of detection, and then its ORF5 gene was compared with modified live virus （MLV） vaccine strain to figure out the influence of vaccine on disease. Between four positive samples＇ sequence, two of them were 99.9%-100% similar to MLV vaccine strain and two other samples were 88.6%-92.7% similar. Similarity rate of the sequences between the vaccine and virus from aborted fetuses are very crucial, because it implies that abortion in swine can be made due to the usage of vaccine not only by the infection of field virus, and if MLV vaccine actually do have an impact on the infection, usage of the vaccine should be reconsidered.
文摘CD （Cryptocarya densiflora） Blume has traditionally been used as an herbal medicine. In this study, the effects of CDEE （CD ethanol extract） on inflammation were investigated in LPS （lipopolysaccharide）-stimulated mouse RAW264.7 macrophages. We investigated the effects of CDEE on the production of NO, PGE2 interleukin （IL）-6, IL-1β, and tumor necrosis factor （TNF）-α in LPS-stimulated RAW264.7 cells. We measured the mRNA or protein expression of the pro-inflammatory mediators induced by CDEE in LPS-stimulated RAW264.7 cells. We explored the expression of Nrf-2, heme oxygenase （HO）-I and NADPH-quinone oxidoreductase （NQO）-I to elucidate the antioxidative mechanisms. CDEE significantly inhibited the production of NO, PGE2, IL-6, IL-1β and TNF-α in LPS-stimulated RAW264.7 cells. CDEE suppressed the mRNA or protein expression of iNOS, COX-2, and the MAPKs with a reduction in the translocation of NF-κB in LPS-stimulated RAW264.7 cells. In addition, CDEE significantly increased the expression of HO-I and NQO-1 with an increase in the translocation of Nrf-2 into the nucleus. These results indicate that CDEE inhibits the LPS-induced inflammatory and oxidative responses via suppression of NF-κB activation and the enhancement of Nrf2 activation. We suggest that CDEE may be therapeutic for treating inflammatory diseases.
基金funded by grants from the National Research Foundation of Korea(NRF)grant funded by the Korea government(2018M3A9H4055203 and 2023R1A2C2003679)from the Korea Health Technology R&D Project through the Korea Health Industry Development Institute(KHIDI),funded by the Ministry of Health&Welfare,Republic of Korea(HV23C1857)from KRIBB Research Initiative Program(KGM9942421).
文摘Influenza,a highly contagious respiratory infectious disease caused by an influenza virus,is a threat to public health worldwide.Avian influenza viruses(AIVs)have the potential to cause the next pandemic by crossing the species barrier through mutation of viral genome.Here,we investigated the pathogenicity of AIVs obtained from South Korea and Mongolia during 2018–2019 by measuring viral titers in the lungs and extrapulmonary organs of mouse models.In addition,we assessed the pathogenicity of AIVs in ferret models.Moreover,we compared the ability of viruses to replicate in mammalian cells,as well as the receptor-binding preferences of AIV isolates.Genetic analyses were finally performed to identify the genetic relationships and amino acid substitutions between viral proteins during mammalian adaptation.Of the 24 AIV isolates tested,A/Mallard/South Korea/KNU2019-34/2019(KNU19-34;H1N1)caused severe bodyweight loss and high mortality in mice.The virus replicated in the lungs,kidneys,and heart.Importantly,KNU19-34-infected ferrets showed high viral loads in both nasal washes and lungs.KNU19-34 replicated rapidly in A549 and bound preferentially to human likeα2,6-linked sialic acids rather than to avian-likeα2,3-linked sialic acids,similar to the pandemic A/California/04/2009(H1N1)strain.Gene segments of KNU19-34 were distributed in Egypt and Asia lineages from 2015 to 2018,and the virus had several amino acid substitutions compared to H1N1 AIV isolates that were non-pathogenic in mice.Collectively,the data suggest that KNU19-34 has zoonotic potential and the possibility of new mutations responsible for mammalian adaptation.
基金supported by Grants from the C1 Gas Refinery Program,funded by the Ministry of Science and ICT(NRF-2018M3D3A1A01056181)the Enzyme engineering for next generation biorefinery(NRF-2022M3J5A1056169 and NRF-2022M3J5A1085239)from National Research Foundation(NRF)the Korea Research Institute of Bioscience and the Biotechnology(KRIBB)Research Initiative Program.
文摘The bioconversion of 4-hydroxy-2-keto acid derivatives via aldol condensation of formaldehyde and pyruvate has received substantial attention as potential source of chemicals for production of amino acids,hydroxy carboxylic acids,and chiral aldehydes.We developed an environmentally friendly biocatalyst consisting of a novel thermostable class II pyruvate aldolase from Deinococcus radiodurans with maltose-binding protein(MBP-DrADL),which has specific activity of 46.3μmol min-1 mg-1.Surprisingly,MBP-DrADL maintained over 60%of enzyme activity for 4 days at 50 to 65°C,we used MBP-DrADL as the best candidate enzyme to produce 2-keto-4-hydroxybutyrate(2-KHB)from formaldehyde and pyruvate via aldol condensation.The optimum reaction conditions for 2-KHB production were 50°C,pH 8.0,5 mM Mg2+,100 mM formaldehyde,and 200 mM pyruvate.Under these optimized conditions,MBP-DrADL produced 76.5 mM(8.94 g L-1)2-KHB over 60 min with a volumetric productivity of 8.94 g L-1 h-1 and a specific productivity of 357.6 mg mg-enzyme-1 h-1.Furthermore,2-KHB production was improved by continuous addition of substrates,which produced approximately 124.8 mM(14.6 g L-1)of 2-KHB over 60 min with a volumetric productivity and specific productivity of 14.6 g L-1 h-1 and 583.4 mg mg-enzyme-1 h-1,respectively.MBP-DrADL showed the highest specific productivity for 2-KHB production yet reported.Our study provides a highly efficient biocatalyst for the synthesis of 2-KHB and lays the foundation for large-scale production and application of high-value compounds from formaldehyde.
基金supported by the Korea Institute of Toxicology(KIT)Research Program[No.2710008763(KK-2401-01)]the Korea Environmental Industry&Technology Institute(KEITI)through the Core Technology Development Project for Environmental Diseases Prevention and Management[No.2480000072(RS-2021-KE001705)]。
文摘Background:Picropodophllotoxin(PPT),a principal component of Podophyllum hexandrum root,demonstrates various beneficial biological activities in multiple cancer types,including antitumor and antiproliferative properties.Despite its known effects,the specific mechanisms by which PPT induces apoptosis in oral squamous cell carcinoma(OSCC)cells lack full clarification.Aims:This study aimed to evaluate the role of PPT in inducing apoptosis in OSCC cells by targeting signal transducer and activator of transcription 3(STAT3)and to investigate the underlying molecular pathways.Methods:Human OSCC cell lines(HN22 and HSC4)were treated with PPT.Cell viability,colony formation,and apoptotic morphological changes were evaluated.Reactive oxygen species(ROS)generation and mitochondrial function were assessed using tetramethyl rhodamine methyl ester,MitoSOX,and 2′,7′-dichlorodihydrofluorescein diacetate(DCFH-DA)assays following PPT treatment.The expression of apoptosis markers,including cleaved Poly(ADP-Ribose)Polymerase(c-PARP)and other target proteins,was measured using western blotting.ROS involvement was further confirmed using the ROS scavenger N-acetylcysteine(NAC).Results:Treatment with PPT resulted in a substantial reduction in cell viability,a decrease in colony formation capacity,and evident morphological changes in OSCC cells.These effects were dose-and time-dependent,as evidenced by increased expression of c-PARP.PPT-induced apoptosis was mediated by excessive ROS generation,which was almost completely blocked by NAC pretreatment.Conclusions:These findings suggest that PPT may serve as a promising therapeutic agent for treating human oral cancer by inhibiting the STAT3 pathway and inducing ROS-mediated apoptosis.
基金supported by a Korea Innovation Foundation(INNOPOLIS)grant funded by the Korean government(Ministry of Science and ICT)through a science and technology project that opens the future of the region(grant number:2021-DD-UP-0380).
文摘Objectives:Limosilactobacillus reuteri is a beneficial Lactobacillus widely used in foods and supplements to promote overall health.Some studies also suggest it supports skin health and prevents allergies and cardiovascular disease.However,research on its skin-protective effects against photoaging has not been conducted.This study evaluated the potential of culture broths fromthree L.reuteri strains(DS0333,DS0384,and DS0385)to inhibit skin photoaging.Methods:To assess their anti-photoaging potential,the culture broths were examined for antioxidant capacity,melanin inhibition,and collagen synthesis promotion.Radical scavenging activity was tested using 2,2-diphenyl-1-picrylhydrazyl(DPPH)and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS)assays.The biosynthetic activity of melanin and associated protein markers involved in melanogenesis was examined in a B16F10 mouse melanoma model.Type I procollagen synthesis and matrix metalloproteinase-1(MMP-1)inhibition were evaluated in ultraviolet B(UVB)-damaged human dermal fibroblasts(HDFs).Results:The culture broths exhibited concentrationdependent antioxidant activity and significantly suppressed melanin synthesis triggered byα-melanocyte-stimulating hormone(α-MSH).Transcription factors involved inmelanogenesis,namelymicrophthalmia-associated transcription factor(MITF),tyrosinase-related protein 1(TRP-1),and 2(TRP-2),were significantly downregulated following treatment.Treatment with culture broths also enhanced type I procollagen production and inhibited MMP-1 activity and protein expression in UVB-exposed HDFs.Among the strains,DS0333 demonstrated the strongest efficacy and was further investigated.It enhanced the proliferation of skin cells and attenuated the levels of age-associated markers such as MMP-1,mitogen-activated protein kinase(MAPK),and activator protein 1(AP-1).High-performance liquid chromatography(HPLC)analysis identified phenyllactic acid(PLA)as the predominant active compound.Conclusions:These results indicate that DS0333 culture broth exhibits strong anti-aging effects and can be applied in functional cosmetics aimed at promoting skin health.
基金supported by the National Research Foundation of Korea(NRF)funded by the government of the Republic of Korea[2023R1A2C1004955]the Technology Innovation Program funded by the Ministry of Trade,Industry&Energy(Korea)(20009707)+1 种基金the Basic Science Research Program through the National Research Foundation of Korea(NRF)funded by the Ministry of Education(2020R1A6A3A01099056)the Korea Institute for Advancement of Technology funded by the Ministry of Trade,Industry and Energy(P0025489).
文摘Infiltration and activation of peripheral immune cells are critical in the progression of multiple sclerosis and its experimental animal model,experimental autoimmune encephalomyelitis(EAE).This study investigates the role of high mobility group box 1(HMGB1)in oligodendrocyte precursor cells(OPCs)in modulating pathogenic T cells infiltrating the central nervous system through the blood-brain barrier(BBB)by using OPC-specific HMGB1 knockout(KO)mice.We found that HMGB1 released from OPCs promotes BBB disruption,subsequently allowing increased immune cell infiltration.The migration of CD4+T cells isolated from EAE-induced mice was enhanced when co-cultured with OPCs compared to oligodendrocytes(OLs).OPC-specific HMGB1 KO mice exhibited lower BBB permeability and reduced immune cell infiltration into the CNS,leading to less damage to the myelin sheath and mitigated EAE progression.CD4+T cell migration was also reduced when co-cultured with HMGB1 knock-out OPCs.Our findings reveal that HMGB1 secretion from OPCs is crucial for regulating immune cell infiltration and provides insights into the immunomodulatory function of OPCs in autoimmune diseases.
基金Supported by The 21st Century Frontier Program in the Functional Human Genome Project, No. HGM0200934the International Collaboration Program of Science and Technology, No. FGM0600914the Ministry of Education, Science and Technology, and the KRIBB Research Initiative Program Grant, No. KGM3320911, South Korea
文摘Long term hepatitis B virus (HBV) infection is a major risk factor in pathogenesis of chronic liver diseases,including hepatocellular carcinoma (HCC). The HBV encod-ed proteins,hepatitis B virus X protein and preS,appear to contribute importantly to the pathogenesis of HCC. Both are associated with oxidative stress,which can damage cellular molecules like lipids,proteins,and DNA during chronic infection. Chronic alcohol use is another important factor that contributes to oxidative stress in the liver. Previous studies reported that treatment with antioxidants,such as curcumin,silymarin,green tea,and vitamins C and E,can protect DNA from damage and regulate liver pathogenesis-related cascades by reducing reactive oxygen species. This review summarizes some of the relationships between oxidative stress and liver pathogenesis,focusing upon HBV and alcohol,and suggests antioxidant therapeutic approaches.
基金Supported by The 21st century Frontier Program in the Functional Human Genome Project,No.HGM0200934the International Collaboration Program of Science and Technology,No. FGM0600914+1 种基金the Research Program for New Drug Target Discovery Grant from the Ministry of Education,Science & Technology,No.NBM3300711the KRIBB Research Initiative Program Grant,No.KGM3320911
文摘AIM:To investigate the role of hepatitis B virus X-protein(HBx)-induced reactive oxygen species(ROS)on liver carcinogenesis in HBx transgenic mice and HepG2-HBx cells.METHODS:Cell growth rate was analyzed,and through western blotting,mitogenic signaling was observed.Endogenous ROS from wild and HBx transgenic mice and HepG2-Mock and HBx cells were assayed by FACS-calibur.Identification of oxidized and reduced phosphatase and tensin homolog(PTEN)was analyzed through N-ethylmaleimide alkylation,nonreducing electrophoresis.RESULTS:We observed that the cell-proliferation-related phosphoinositide 3-kinase/Akt pathway is activated by HBx in vivo and in vitro.Increased ROS were detected by HBx.Tumor suppressor PTEN,via dephosphorylation of Akt,was oxidized and inactivated by increased ROS.Increased oxidized PTEN activated the mitogenic pathway through over-activated Akt.However,treatment with ROS scavenger N-acetyl cysteine can reverse PTEN to a reduced form.Endogenously produced ROS also stimulated HBx expression.CONCLUSION:HBx induced ROS promoted Akt pathways via oxidized inactive PTEN.HBx and ROS maintained a positive regulatory loop,which aggravated carcinogenesis.
基金supported by the China Agriculture Research System (CARS-10,Sweetpotato)the Agricultural Science and Technology Innovation Program of Jiangsu Province,China (CX(13)2032)the China-Korea Young Scientist Exchange Program
文摘Soil salinity causes the negative effects on the growth and yield of crops. In this study, two sweet potato （Ipomoea batatas L.） cultivars, Xushu 28 （X-28） and Okinawa 100 （O-100）, were examined under 50 and 100 mmol L-1 NaCI stress. X-28 cultivar is relatively high salt tolerant than O-100 cultivar. Interestingly, real-time quantitative polymerase chain reaction （RT-qPCR） results indicated that sweet potato high-affinity K^＋ transporter 1 （IbHKT1） gene expression was highly induced by 50 and 100 mmol L-1 NaCI stress in the stems of X-28 cultivar than in those of O-100 cultivar, but only slightly induced by these stresses in the leaves and fibrous roots in both cultivars. To characterize the function of IbHKT1 transporter, we performed ion-flux analysis in tobacco transient system and yeast complementation. Tobacco transient assay showed that IbHKT1 could uptake sodium （Na^＋）. Yeast complementation assay showed that IbHKT1 could take up K^＋ in 50 mmol L^-1 K^＋ medium without the presence of NaCI. Moreover, Na^＋ uptake significantly increased in yeast overexpressing IbHKTI. These results showed that IbHKT1 transporter could have K^＋-Na^＋ symport function in yeast. Therefore, the modes of action of IbHKT1 in transgenic yeast could differ from the mode of action of the other HKT1 transporters in class I. Potentially, IbHKT1 could be used to improve the salt tolerance nature in sweet potato.
基金supported by Grants from the R&D Convergence Program of National Research Council of Science & Technology (No. CAP-16-02-KIST)the National Research Foundation of Korea (No. NRF-2016M3A9B6918584)
文摘Chikungunya fever is a vector-borne viral disease transmitted to humans by chikungunya virus(CHIKV)-infected mosquitoes.There have been many outbreaks of CHIKV infection worldwide,and the virus poses ongoing risks to global health.To prevent and control CHIKV infection,it is important to improve the current CHIKV diagnostic approaches to allow for the detection of low CHIKV concentrations and to correctly distinguish CHIKV infections from those due to other mosquito-transmitted viruses,including dengue virus(DENV),Japanese encephalitis virus(JEV),and Zika virus(ZIKV).Here,we produced monoclonal antibodies(mAbs)against the CHIKV envelope 2 protein(CHIKV-E2)and compared their sensitivity and specificity with commercially available mAbs using enzyme-linked immunosorbent assays(ELISA).Two anti-CHIKV-E2 mAbs,19-1 and 21-1,showed higher binding affinities to CHIKV-E2 protein than the commercial mAbs did.In particular,the 19-1 mAb had the strongest binding affinity to inactivated CHIKV.Moreover,the 19-1 mAb had very little cross-reactivity with other mosquito-borne viruses,such as ZIKV,JEV,and DENV.These results suggest that the newly produced anti-CHIKV-E2 mAb,19-1,could he used for CHIKV diagnostic approaches.
基金supported by the research fund of Chungnam National University,Republic of Koreasupported by the Li Kashing Foundation at Shantou University Medical College,China
文摘Clusterin, a protein associated with multiple functions, is expressed in a wide variety of mammalian tissues. Although clusterin is known to be involved in neurodegenerative diseases, ageing, and tumorigenesis, a detailed analysis of the consequences of gain- or loss-of- function approaches has yet to be performed to understand the underlying mechanisms of clusterin functions. Since clusterin levels change in neurological diseases, it is likely that clusterin contributes to cell death and degeneration in general. Zebrafish was investigated as a model system to study human diseases. During development, zebrafish clusterin was expressed in the notochord and nervous system. Embryonic overexpression of clusterin by mRNA microinjection did not affect axis formation, whereas its knock-down by anti-sense morpholino treatment resulted in neuronal cell death. To analyze the function of clusterin in neurodegeneration, a transgenic zebrafish was investigated, in which nitroreductase expression is regulated under the control of a neuron-specifc huC promoter which is active between the stages of early neuronal precursors and mature neurons. Nitroreductase turns metronidazole into a cytotoxic agent that induces cell death within 12 h. After metronidazole treatment, transgenic zebrafish showed neuron-specific cell death. Interestingly, we also observed a dramatic induction of clusterin expression in the brain and spinal cord in these fish, suggesting a direct or indirect role of clusterin in neuronal cell death and thus, more generally, in neurodegeneration.
基金supported by the Ministry of EducationScience and Technology(No.2021M3A9A1096894)+1 种基金Republic of Korea and the KRIBB Research Initiative Program(KGM4252223)Korea Research Institute of Bioscience and Biotechnology,Republic of Korea。
文摘Background Anethole(AN)is an organic antioxidant compound with a benzene ring and is expected to have a positive impact on early embryogenesis in mammals.However,no study has examined the effect of AN on porcine embryonic development.Therefore,we investigated the effect of AN on the development of porcine embryos and the underlying mechanism.Results We cultured porcine in vitro-fertilized embryos in medium with AN(0,0.3,0.5,and 1 mg/mL)for 6 d.AN at 0.5 mg/mL significantly increased the blastocyst formation rate,trophectoderm cell number,and cellular survival rate compared to the control.AN-supplemented embryos exhibited significantly lower reactive oxygen species levels and higher glutathione levels than the control.Moreover,AN significantly improved the quantity of mitochondria and mitochondrial membrane potential,and increased the lipid droplet,fatty acid,and ATP levels.Interestingly,the levels of proteins and genes related to the sonic hedgehog(SHH)signaling pathway were significantly increased by AN.Conclusions These results revealed that AN improved the developmental competence of porcine preimplantation embryos by activating SHH signaling against oxidative stress and could be used for large-scale production of high-quality porcine embryos.
基金supported by the 2013 Inje University Research Grant
文摘Experimental stroke research commonly employs focal cerebral ischemic rat models （Bederson et al., 1986a; Longa et al., 1989）. In human patients, ischemic stroke typically results from thrombotic or embolic occlusion of a major cerebral artery, usually the mid- dle cerebral artery （MCA）. Experimental focal cerebral ischemia models have been employed to mimic human stroke （Durukan and Tatlisumak, 2007）. Rodent models of focal cerebral ischemia that do not require craniotomy have been developed using intraluminal suture occlusion of the MCA （MCA occlusion, MCAO） （Rosamond et al., 2008）. Furthermore, mouse MCAO models have been wide- ly used and extended to genetic studies of cell death or recovery mechanisms （Liu and McCullough, 2011）. Genetically engineered mouse stroke models are particularly useful for evaluation of isch- emic pathophysiology and the design of new prophylactic, neuro- protective, and therapeutic agents and interventions （Armstead et al., 2010）. During the past two decades, MCAO surgical techniques have been developed that do not reveal surgical techniques for mouse MCAO model engineering. Therefore, we compared MCAO surgical methods in rats and mice.
基金Supported by The World Class Institute(WCI) Program of the Na-tional Research Foundation of Korea(NRF),No.WCM0101222Ministry of Education,Science and Technology of Korea(MEST),the National R and D Program for Cancer Control,Ministry of Health and Welfare,South Korea,No.BCM0061213KRIBB Research Initiative Program Grant,No.KGM3141312
文摘AIM:To investigate the effect of methylsulfonylmethane(MSM), recently reported to have anti-cancer effects, in liver cancer cells and transgenic mice. METHODS: Three liver cancer cell lines, HepG2, Huh7-Mock and Huh7-H-rasG12V, were used. Cell growth was measured by Cell Counting Kit-8 and soft agar assay. Western blot analysis was used to detect caspases, poly(ADP-ribose) polymerase(PARP), and B-cell lymphoma2(Bcl-2) expressions. For in vivo study, we administered MSM to H-ras12V transgenic mice for 3 mo. RESULTS: MSM decreased the growth of HepG2, Huh7-Mock and Huh7-H-rasG12V cells in a dose-dependent manner. That was correlated with significantly increased apoptosis and reduced cell numbers in MSM treated cells. Cleaved caspase-8, cleaved caspase-3 and cleaved PARP were remarkably increased in the liver cancer cells treated with 500 mmol/L of MSM; however, Bcl-2 was slightly decreased in 500 mmol/L. Liver tumor development was greatly inhibited in the H-ras12V transgenic mice treated with MSM, compared to control, by showing reduced tumor size and number. Cleaved PARP was significantly increased in non-tumor treated with MSM compared to control. CONCLUSION: Liver injury was also significantly attenuated in the mice treated with MSM. Taken together, all the results suggest that MSM has anti-cancer effects through inducing apoptosis in liver cancer.
