The fine-tuned expression dynamics of the effector genes are pivotal for the transition from vegetative growth to host colonization of pathogenic filamentous fungi.However,mechanisms underlying the dynamic regulation ...The fine-tuned expression dynamics of the effector genes are pivotal for the transition from vegetative growth to host colonization of pathogenic filamentous fungi.However,mechanisms underlying the dynamic regulation of these genes remain largely unknown.Here,through comparative transcriptome and chromatin immunoprecipitation sequencing(ChIP-seq)analyses of the methyltransferase PoKmt6 in rice blast fungus Pyricularia oryzae(syn.Magnaporthe oryzae),we found that PoKmt6-mediated H3K27me3 deposition was enriched mainly at fast-evolving regions and contributed to the silencing of a subset of secreted proteins(SP)and transposable element(TE)families during the vegetative growth of P.oryzae.Intriguingly,we observed that a group of SP genes,which were depleted of H3K27me3 modification,could also be silenced via the H3K27me3-mediated repression of the nearby TEs.In conclusion,our results indicate that H3K27me3 modification mediated by PoKmt6 regulates the expression of some SP genes in fast-evolving regions through the suppression of nearby TEs.展开更多
Objective Obstructive sleep apnea (OSA) is closely related to obesity, insulin resistance and inflammation. Secreted frizzled-related protein 5 (SFRP5) is a recently discovered adipokine. It is involved in insulin res...Objective Obstructive sleep apnea (OSA) is closely related to obesity, insulin resistance and inflammation. Secreted frizzled-related protein 5 (SFRP5) is a recently discovered adipokine. It is involved in insulin resistance and inflammation in obesity. This study aimed at evaluating the association between SFRP5and sleeping characteristics as well as biochemical parameters of OSA patients.Methods This was a prospective case control study. Nondiabetic OSA patients and controls were consecutively recruited and divided into three groups: OSA group, apnea–hypopnea Index (AHI)≥5/h; healthy controls with normal body mass index (BMI); obese controls without OSA, and BMI > 24.0 kg/m2. All participants underwent polysomnography (PSG). Plasma SFRP5 was examined using enzyme-linked immunosorbent assay (ELISA). Blood biochemical examinations, including fasting blood glucose (FBG), lipid profile, hypersensitive Creactive protein (hsCRP), were performed early in the morning after PSG. Patients with severe OSA were treated with nasal continuous positive airway pressure (nCPAP), and plasma SFRP5 was repeatedly measured for comparison.Results Sixty-eight subjects were enrolled in the study, including 38 patients of OSA, whose medium AHI was 58.70 /h (36.63, 71.15), 20 obese controls, and 10 healthy controls. The plasma SFRP5 level of OSA patients was not significantly different from that of healthy controls or obese controls. In OSA patients, SFRP5 level correlated positively with triglyceride level (r=0.447, P=0.005) and negatively with LDL-cholesterol level and HDLcholesterol level (r=?0.472 and P=0.003; r=?0.478 and P=0.002; respectively). SFRP5 level was not found correlating with FBG, AHI, or any of nocturnal hypoxia parameters. After overnight nCPAP treatment, plasma SFRP5 levels of OSA patients did not change significantly (t=1.557, P = 0.148) compared to that of pretreatment.Conclusions In nondiabetic OSA patients, plasma SFRP5 is associated with the lipid profile. However,no correlation was observed between SFRP5 and FBG or sleep parameters. The SFRP5 level of OSA patients did not differ from that of non-OSA individuals in our study.展开更多
BACKGROUND Sepsis is a major medical challenge.Magnolol is an active constituent of Houpu that improves tissue function and exerts strong anti-endotoxin and anti-inflammatory effects,but the mechanism by which it redu...BACKGROUND Sepsis is a major medical challenge.Magnolol is an active constituent of Houpu that improves tissue function and exerts strong anti-endotoxin and anti-inflammatory effects,but the mechanism by which it reduces intestinal inflammation in sepsis is yet unclear.AIM To assess the protective effect of magnolol on intestinal mucosal epithelial cells in sepsis and elucidate the underlying mechanisms.METHODS Enzyme-linked immunosorbent assay was used to measure tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),IL-6,and regulated on activation,normal T-cell expressed and secreted(RANTES)levels in serum and ileal tissue in animal studies.The histopathological changes of the ileal mucosa in different groups were observed under a microscope.Cell Counting Kit-8 and cell permeability assays were used to determine the concentration of drug-containing serum that did not affect the activity of Caco2 cells but inhibited lipopolysaccharide(LPS)-induced decrease in permeability.Immunofluorescence and Western blot assays were used to detect the levels of RANTES,inhibitor of nuclear factor kappa-B kinaseβ(IKKβ),phosphorylated IKKβ(p-IKKβ),inhibitor of nuclear factor kappa-B kinaseα(IκBα),p65,and p-p65 proteins in different groups in vitro.RESULTS In rats treated with LPS by intravenous tail injection in the presence or absence of magnolol,magnolol inhibited the expression of proinflammatory cytokines,IL-1β,IL-6,and TNF-αin a dose-dependent manner.In addition,magnolol suppressed the production of RANTES in LPS-stimulated sepsis rats.Moreover,in vitro studies suggested that magnolol inhibited the increase of p65 nucleation,thereby markedly downregulating the production of the phosphorylated form of IKKβin LPS-treated Caco2 cells.Specifically,magnolol inhibited the translocation of the transcription factor nuclear factor-kappa B(NF-κB)from the cytosol into the nucleus and down-regulated the expression level of the chemokine RANTES in LPS-stimulated Caco2 cells.CONCLUSION Magnolol down-regulates RANTES levels by inhibiting the LPS/NF-κB signaling pathways,thereby suppressing IL-1β,IL-6,and TNF-αexpression to alleviate the mucosal barrier dysfunction in sepsis.展开更多
Computer based software such as the SignalP v3.0, TargetP v1.01, big-PI predictor and TMHMM v2.0 were combined topredict the signal peptides, and the signal peptide-dependent secreted proteins among the 6 700 ORFs in ...Computer based software such as the SignalP v3.0, TargetP v1.01, big-PI predictor and TMHMM v2.0 were combined topredict the signal peptides, and the signal peptide-dependent secreted proteins among the 6 700 ORFs in genome ofSaccharomyces cerevisiae. The results showed that 163 proteins were the secreted ones containing signal peptides, andthey were secreted via Sec pathway. Among the 163 predicted secreted proteins, the signal peptides of 47 secretedproteins included only the H-domain and C-domain, without N-domain, but the signal peptides of other 116 secretedproteins included all the three domains. There were differences in the constitution of signal peptides between the secretedproteins of S. cerevisiae and of Candida albicans, but the length and amino acids types of their signal peptides weresimilar in general. Few of the same signal peptides occurred in the secreted proteins of S. cerevisiae genome, and thehomology could be compared among the secreted proteins with the same signal peptides. The BLAST 2 SEQUENECESand CLUSTAL W were used to align the two protein sequences and multi-protein sequences, respectively. The alignmentresult indicated that homology of these sequences with the same signal peptide was very highly conservative in aminoacid of complete gene. The effect of the signal peptides in S. cerevisia on expression of foreign eukaryotic secretedproteins is discussed in this paper.展开更多
BACKGROUND Aberrant methylation in DNA regulatory regions could downregulate tumor suppressor genes without changing the sequences.However,our knowledge of secreted protein acidic and rich in cysteine(SPARC)and its ab...BACKGROUND Aberrant methylation in DNA regulatory regions could downregulate tumor suppressor genes without changing the sequences.However,our knowledge of secreted protein acidic and rich in cysteine(SPARC)and its aberrant methylation in gastric cancer(GC)is still inadequate.In the present research,we performed fundamental research to clarify the precise function of methylation on SPARC and its significance in GC.AIM To investigate promoter methylation and the effects of the SPARC gene in GC cells and tissues and to evaluate its clinical significance.METHODS Plasmids that overexpressed the SPARC gene were transfected into human GC BGC-823 cells;non-transfected cells were used as a control group(NC group).Quantitative real-time polymerase chain reaction and western blotting(WB)were then used to detect the expression of SPARC.Methylation-specific polymerase chain reaction was executed to analyze the gene promoter methylation status.Cell viability was measured by the cell counting kit-8 assay.The migration and invasion ability of cells were detected by scratch assays and transwell chamber assays,respectively.Cell cycle events and apoptosis were observed with a flow cytometer.RESULTS The expression of SPARC mRNA in GC tissues and cells was significantly lower and showed differing degrees of hypermethylation,respectively,than that in normal adjacent tissues and control cells.Treatment with 5-Aza-2’-deoxycytidine(5-Aza-Cdr)was able to restore the expression of SPARC and reverse promoter hypermethylation.Overexpression of the SPARC gene significantly inhibited proliferation,migration,and invasion of GC cells,while also causing cell cycle arrest and apoptosis;the NC group exhibited the opposite effects.CONCLUSION This study demonstrated that SPARC could function as a tumor suppressor and might be silenced by promoter hypermethylation.Furthermore,in GC cells,SPARC inhibited migration,invasion,and proliferation,caused cell cycle arrest at the G0/G1 phase,and promoted apoptosis.展开更多
BACKGROUND: It has been proved by many experimental studies from the aspects of morphology and immunocytochemistry in recent years that bone marrow stromal cells (BMSCs) can in vitro induce and differentiate into the ...BACKGROUND: It has been proved by many experimental studies from the aspects of morphology and immunocytochemistry in recent years that bone marrow stromal cells (BMSCs) can in vitro induce and differentiate into the cells possessing the properties of nerve cells. But the functions of BMSCs-derived neural stem cells(NSCs) and the differentiated neuron-like cells are still unclear. OBJECTIVE: To observe whether bone marrow-derived NSCs can secrete norepinephrine (NE) under the condition of in vitro culture, induce and differentiation, and analyze the biochemical properties of BMSCs-derived NSCs. DESIGN: A non-randomized and controlled experimental observation. SETTING: Institute of Neuromedicine of Chinese PLA, Zhujiang Hospital, Southern Medical University. MATERIALS: This experiment was carried out in the Institute of Neuromedicine of Chinese PLA, Zhujiang Hospital, Southern Medical University. The bone marrow used in the experiment was collected from 1.5- month-old healthy New Zealand white rabbits. METHODS: This experiment was carried out in the Institute of Neuromedicine of Chinese PLA, Zhujiang Hospital, Southern Medical University. The bone marrow used in the experiment was collected from 1.5 month-old healthy New Zealand white rabbits. BMSCs of rabbits were isolated and performed in vitro culture, induce and differentiation with culture medium of NSCs and differentiation-inducing factor, then identified with immunocytochemical method. Experimental grouping: ①Negative control group: L-02 hepatic cell and RPMI1640 culture medium were used. ② Background culture group: Only culture medium of NSCs as culture solution was added into BMSCs to perform culture, and 0.1 volume fraction of imported fetal bovine serum was supplemented 72 hours later. ③Differentiation inducing factor group: After culture for 72 hours, retinoic acid and glial cell line-derived neurotrophic factors were added in the culture medium of BMSCs and NSCs as corresponding inducing factors. The level of NE in each group was detected on the day of culture and 5, 7, 14 and 20 days after culture with high performance liquid chromatography(HPLC). The procedure was conducted 3 times in each group.Standard working curve was made according to the corresponding relationship of NE concentration and peak area. The concentration of NE every 1×107 cells was calculated according to standard curve and cell counting. MAIN OUTCOME MEASURES: The level of NE of cultured cells was detected with HPLC; immunocytochemistrical identification of Nestin and neuron specific nuclear protein was performed. RESULTS: ① On the 14th day after cell culture, BMSCs turned into magnus and round cells which presented Nestin-positive antigen, then changed into neuron-like cells with long processus and presented neuron specific nuclear protein -positive antigen at the 20th day following culture. ② The ratio of NE concentration and peak area has good linear relationship, and regression equation was Y=1.168 36+0.000 272 8X,r=0.998 4.Coefficient variation (CV) was < 5% and the recovery rate was 92.39%(Y referred to concentration and X was peak area).③NE was well detached within 10 minutes under the condition of this experiment. ④ NE was detected in NSCs and their culture mediums, which were cultured for 7, 14 and 20 days respectively, but no NE in BMSCs, NSCs-free culture medium and L-02 hepatic cell which were as negative control under the HPLC examination. Analysis of variance showed that the level of NE gradually increased following the elongation of culture time (P < 0.01). No significant difference in the level of NE existed at the same time between differentiation inducing factor group and basic culture group(P > 0.05). CONCLUSION: BMSCs of rabbits can proliferate in vitro and express Nestin antigen; They can differentiate into neuron-like cells, express specific neucleoprotein of mature neurons, synthesize and secrete NE as a kind of neurotransmitter.展开更多
Protein secretion plays an important role in bacterial lifestyles. In Gram-negative bacteria, a wide range of proteins are secreted to modulate the interactions of bacteria with their environments and other bacteria v...Protein secretion plays an important role in bacterial lifestyles. In Gram-negative bacteria, a wide range of proteins are secreted to modulate the interactions of bacteria with their environments and other bacteria via various secretion systems. These proteins are essential for the virulence of bacteria, so it is crucial to study them for the pathogenesis of diseases and the development of drugs. Using amino acid composition (AAC), position-specific scoring matrix (PSSM) and N-terminal signal peptides, two different substitution models are firstly constructed to transform protein sequences into numerical vectors. Then, based on support vector machine (SVM) and the “one to one”?algorithm, a hybrid multi-classifier named SecretP v.2.2 is proposed to rapidly and accurately?distinguish different types of Gram-negative?bacterial secreted proteins. When performed on the same test set for a comparison with other methods, SecretP v.2.2 gets the highest total sensitivity of 93.60%. A public independent dataset is used to further test the power of SecretP v.2.2 for predicting NCSPs, it also yields satisfactory results.展开更多
In order to analyze the in vivo expression of Candida albicans secreted aspartyl pro- teinases (SAP) in human vaginal infection, the vaginal secretion from 29 human subjects was col- lected by vaginal swab, and the ex...In order to analyze the in vivo expression of Candida albicans secreted aspartyl pro- teinases (SAP) in human vaginal infection, the vaginal secretion from 29 human subjects was col- lected by vaginal swab, and the expression of SAP1–SAP6 was detected by reverse-transcriptase po- lymerase chain reaction using specific primer sets. It was found that Sap2 and Sap5 were the most common genes expressed during infection; Sap3 and Sap4 were detected in all subjects and all 6 SAP genes were simultaneously expressed in some patients with vaginal candidiasis. It was suggested that the SAP family is expressed by Candida albicans during infection in human and that Candida albi- cans infection is associated with the differential expression of individual SAP genes which may be involved in the pathogenesis of vaginal candidiasis.展开更多
BACKGROUND Previously,we have successfully constructed replication-competent hepatitis B virus(HBV)vectors by uncoupling the P open reading frame(ORF)from the preC/C ORF to carefully design the transgene insertion sit...BACKGROUND Previously,we have successfully constructed replication-competent hepatitis B virus(HBV)vectors by uncoupling the P open reading frame(ORF)from the preC/C ORF to carefully design the transgene insertion site to overcome the compact organization of the HBV genome and maintain HBV replication competence.Consequently,the replication-competent HBV vectors carrying foreign genes,including pCH-BsdR,carrying blasticidin resistance gene(399 bp),and pCH-hrGFP,carrying humanized renilla green fluorescent protein gene(720 bp),were successfully obtained.However,the replication efficiency of the former is higher but it is tedious to use,while that of the latter is poor and cannot be quantified.Hence,we need to search for a new reporter gene that is convenient and quantifiable for further research.AIM To establish a helpful tool for intracellular HBV replication and anti-viral drugs screening studies.METHODS We utilized the replication-competent HBV viral vectors constructed by our laboratory,combined with the secreted luciferase reporter gene,to construct replication-competent HBV vectors expressing the reporter gene secretory Nanoluc Luciferase(SecNluc).HepG2.TA2-7 cells were transfected with this vector to obtain cell lines with stably secreted HBV particles carrying secNluc reporter gene.RESULTS The replication-competent HBV vector carrying the SecNluc reporter gene pCHsNLuc could produce all major viral RNAs and a full set of envelope proteins and achieve high-level secreted luciferase expression.HBV replication intermediates could be produced from this vector.Via transfection with pTRE-sNLuc and selection by hygromycin,we obtained isolated cell clones,named HBV-NLuc-35 cells,which could secrete secNLuc recombinant viruses,and were sensitive to existing anti-HBV drugs.Using differentiated HepaRG cells,it was verified that recombinant HBV possessed infectivity.CONCLUSION Our research demonstrated that a replication-competent HBV vector carrying a secreted luciferase transgene possesses replication and expression ability,and the established HBV replication and expression cell lines could stably secrete viral particles carrying secNluc reporter gene.More importantly,the cell line and the secreted recombinant viral particles could be used to trace HBV replication or infection.展开更多
BACKGROUND Secreted protein acidic and rich in cysteine(SPARC)is an extracellular matrixassociated protein.Studies have revealed that SPARC is involved in the cell interaction and function including proliferation,diff...BACKGROUND Secreted protein acidic and rich in cysteine(SPARC)is an extracellular matrixassociated protein.Studies have revealed that SPARC is involved in the cell interaction and function including proliferation,differentiation,and apoptosis.However,the role of SPARC in cancer is controversial,as it was reported as the promoter or suppressor in different cancers.Further,the role of SPARC in lymphoma is unclear.AIM To identify the expression and significance of SPARC in lymphoma,especially in diffuse large B-cell lymphoma(DLBCL).METHODS The expression analysis of SPARC in different cancers was evaluated with Oncomine.The Brune,Eckerle,Piccaluga,Basso,Compagno,Alizadeh,and Rosenwald datasets were included to evaluate the mRNA expression of SPARC in lymphoma.The Cancer Genome Atlas(TCGA)-DLBCL was used to analyze the diagnostic value of SPARC in DLBCL.The Compagno and Brune DLBCL datasets were used for validation.Then,the diagnostic value was evaluated with the receiver operating characteristic(ROC)curve.The Kaplan-Meier plot was conducted with TCGA-DLBCL,and the ROC analysis was performed based on the survival time.Further,the overall survival analysis based on the level of SPARC expression was performed with the GSE4475 and E-TABM-346.The Gene Set Enrichment Analyses(GSEA)was performed to make the underlying mechanism-regulatory networks.RESULTS The pan-cancer analysis of SPARC showed that SPARC was highly expressed in the brain and central nervous system,breast,colon,esophagus,stomach,head and neck,pancreas,and sarcoma,especially in lymphoma.The overexpression of SPARC in lymphoma,especially DLBCL,was confirmed in several datasets.The ROC analysis revealed that SPARC was a valuable diagnostic biomarker.More importantly,compared with DLBCL patients with low SPARC expression,those with higher SPARC expression represented a higher overall survival rate.The ROC analysis showed that SPARC was a favorable prognostic biomarker for DLBCL.Results of the GSEA confirmed that the high expression of SPARC was closely associated with focal adhesion,extracellular matrix receptor interaction,and leukocyte transendothelial migration,which suggested that SPARC may be involved in the regulation of epithelial-mesenchymal transition,KRAS,and myogenesis in DLBCL.CONCLUSION SPARC was highly expressed in DLBCL,and the overexpression of SPARC showed sound diagnostic value.More interestingly,the overexpression of SPARC might be a favorable prognostic biomarker for DLBCL,suggesting that SPARC might be an inducible factor in the development of DLBCL,and inducible SPARC was negative in some oncogenic pathways.All the evidence suggested that inducible SPARC might be a good diagnostic and prognostic biomarker for DLBCL.展开更多
The structure similarity of secreted proteins in rice blast fungus Magnaporthe oryzae and its host Oryza sativa was analyzed. One thousand two hundred and forty one proteins were predicted as secreted proteins using f...The structure similarity of secreted proteins in rice blast fungus Magnaporthe oryzae and its host Oryza sativa was analyzed. One thousand two hundred and forty one proteins were predicted as secreted proteins using four algorithms based on 11 074 proteins in genome of M. oryzae. One hundred and forty six secreted proteins( 11. 8% of M. oryzae secretome) were aligned with 116 rice proteins( 0. 21% of 56 278 rice proteins) using BLAST search on rice genome. One hundred sixteen rice similar proteins participated in rice cell wall modification( cell wall associated enzymes) and signal transduction( proteases). These results imply that both cell wall involved proteins and signal transduction are probably hijacks pathway between host pants and pathogenic fungi. Because these proteins are highly conserved among fungi and plants,the express patterns of these protein coding genes during the interaction process are valuable to study in detail.展开更多
Corynebacterium pseudotuberculosis is an infectious agent that occurs in small ruminants causing caseous lymphadenitis, and more rarely in humans causing lymphadenitis and pneumonia. The breeding small ruminants have ...Corynebacterium pseudotuberculosis is an infectious agent that occurs in small ruminants causing caseous lymphadenitis, and more rarely in humans causing lymphadenitis and pneumonia. The breeding small ruminants have great economic importance in Brazil. Rural farm workers and veterinary students who acquired this disease suffered from weakening symptoms for weeks, and the identification of the etiological agent was time-consuming and complex. Due to the low prevalence of case records, there is probably no available commercial diagnostic kit for C. pseudotuberculosis infection in humans. This study aimed to describe human seroreactivity to secreted antigens from C. pseudotuberculosis. Reactivity of serum from farm workers (n = 14), individuals who work with the bacillus at laboratory (n = 8) or individuals without contact (n = 25) was tested with secreted proteins from PAT10 strain of C. pseudotuberculosis by Western blotting. Samples of all (100%) farm workers showed reactivity to 31 kDa, 71 kDa and 164 kDa proteins, while laboratory workers showed 87.5%, 62.5 % and 37.5%, and no-contact 20%, 0% and 16%, respectively. All sera recognized the 275 kDa protein. Our data suggest that C. pseudotuberculosis secreted proteins are antigenic in humans and the recognition profiles allowed the identification of individuals with and without prior contact with this bacillus. This is the first paper which describes human reactivity to C. pseudotuberculosis in serum samples of workers in Brazil.展开更多
Phosphorus is one of the most important nutrients required for plant growth and development. While substantial amounts of total phosphorus are present in many soil types, plants are unable to utilize some organic phos...Phosphorus is one of the most important nutrients required for plant growth and development. While substantial amounts of total phosphorus are present in many soil types, plants are unable to utilize some organic phosphorus sources. The main goal of this study was to characterize the spectrum of secreted plant proteins, organic acids and other metabolites that can potentially contribute to utilization of various phosphorus compounds. Our data indicate that the composition of extracellular proteins secreted by plant roots varies depending on the specific source of P in the growth medium. Furthermore, some root-secreted metabolites, such as citrate, appear to be specific to a subset of ecotypes, while tartrate, succinate and oxalate are secreted by a number of A. thaliana ecotypes. We observed secretion of phenolic compounds, such as tannins, and deoxycytidine derivatives. Taken together, while no single secreted polypeptide, organic acid or secondary metabolite can be pinpointed as specific to plant growth in particular phosphorus conditions, our data indicate that A. thaliana ecotypes differ in their physiological responses to the source of phosphorus in the growth medium. Overall, these results suggest that physiological changes in plant responses to nutrient limitation are modulated by interactions between soil phosphorus source and the specific genotype of Arabidopsis plants.展开更多
Background:Cancer-associated fibroblasts(CAFs)play an important role in the induction of chemo-resistance.This study aimed to clarify the mechanism underlying CAF-mediated resistance to two tyrosine kinase inhibitors(...Background:Cancer-associated fibroblasts(CAFs)play an important role in the induction of chemo-resistance.This study aimed to clarify the mechanism underlying CAF-mediated resistance to two tyrosine kinase inhibitors(TKIs),sorafenib and lenvatinib,and to identify a novel therapeutic target for overcoming TKI resistance in hepatocellular carcinoma(HCC).Methods:We performed a systematic integrative analysis of publicly available gene expression datasets and whole-transcriptome sequencing data from 9 pairs of CAFs and para-cancer fibroblasts isolated from human HCC and para-tumor tissues,respectively,to identify key molecules that might induce resistance to TKIs.We then performed in vitro and in vivo experiments to validate selected targets and related mechanisms.The associations of plasma secreted phosphoprotein 1(SPP1)expression levels before sorafenib/lenvatinib treatment with progression-free survival(PFS)and overall survival(OS)of 54 patients with advanced HCC were evaluated using Kaplan-Meier and Cox regression analysis.Results:Bioinformatic analysis identified CAF-derived SPP1 as a candidate molecule driving TKI resistance.SPP1 inhibitors reversed CAF-induced TKI resistance in vitro and in vivo.CAF-derived SPP1 activated rapidly accelerated fibrosarcoma(RAF)/mitogen-activated protein kinase(MAPK)and phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)/mammalian target of rapamycin(mTOR)through the integrin-protein kinase C-alpha(PKCα)signaling pathway and promoted epithelial-to-mesenchymal transition(EMT).A high plasma SPP1 level before TKI treatment was identified as an independent predictor of poor PFS(P=0.026)and OS(P=0.047)in patients with advanced HCC after TKI treatment.Conclusions:CAF-derived SPP1 enhances TKI resistance in HCC via bypass activation of oncogenic signals and EMT promotion.Its inhibition represents a promising therapeutic strategy against TKI resistance inHCC.Moreover,plasma SPP1 level before TKI treatment represents a potential biomarker for treatment response prediction.展开更多
Background and aims:Secreted phosphoprotein 1(SPP1)functions in several physiological processes.The role of SPP1 expression in the prognosis and tumor immunity of hepatocellular carcinoma(HCC)is unknown.The aim of thi...Background and aims:Secreted phosphoprotein 1(SPP1)functions in several physiological processes.The role of SPP1 expression in the prognosis and tumor immunity of hepatocellular carcinoma(HCC)is unknown.The aim of this study was to investigate the expression pattern of SPP1 in HCC and its correlation with prognosis and tumor immunity.Methods:Clinical and gene expression data of The Cancer Genome Atlas-liver hepatocellular carcinoma(LIHC)cohort and 11 other HCC datasets were collected.The Kaplan–Meier method and Cox regression analysis were used to analyze the prognostic value of SPP1.The DESeq2 package in R was used to analyze SPP1-related genes.Gene Ontology analysis and gene set enrichment analysis were used to determine the biological function of SPP1 in HCC.The single sample Gene Set Enrichment Analysis(ssGSEA)method was used to analyze the immune infiltrates of HCC.Illumina human methylation 450 data and level 3 HTSeq-FPKM data from The Cancer Genome Atlas-LIHC were used to analyze the effects of DNA methylation level on SPP1 expression.Results:SPP1 was overexpressed in HCC and correlated with T stage,histological grade,adjacent hepatic tissue inflammation,and vascular invasion in HCC.The analysis of survival rates indicated that high SPP1 levels were associated with poor overall survival in HCC.Functional analysis showed that SPP1 is related to tumor immunity,especially macrophage infiltration.Aberrant demethylation of the promoter region is one of the mechanisms underlying the increase of SPP1 in HCC.Conclusion:Our results indicate that SPP1 is an independent prognostic factor for HCC and is correlated with the clinical features and macrophage infiltration in HCC.展开更多
In the face of increasingly serious environmental pollution,the health of human lung tissues is also facing serious threats.Mogroside IIE(M2E)is the main metabolite of sweetening agents mogrosides from the anti-tussiv...In the face of increasingly serious environmental pollution,the health of human lung tissues is also facing serious threats.Mogroside IIE(M2E)is the main metabolite of sweetening agents mogrosides from the anti-tussive Chinese herbal Siraitia grosvenori.The study elucidated the anti-inflammatory action and molecular mechanism of M2E against acute lung injury(ALI).A lipopolysaccharide(LPS)-induced ALI model was established in mice and MH-S cells were employed to explore the protective mechanism of M2E through the western blotting,co-immunoprecipitation,and quantitative real time-PCR analysis.The results indicated that M2E alleviated LPS-induced lung injury through restraining the activation of secreted phospholipase A2 type IIA(Pla2g2a)-epidermal growth factor receptor(EGFR).The interaction of Pla2g2a and EGFR was identified by co-immunoprecipitation.In addition,M2E protected ALI induced with LPS against inflammatory and damage which were significantly dependent upon the downregulation of AKT and m TOR via the inhibition of Pla2g2a-EGFR.Pla2g2a may represent a potential target for M2E in the improvement of LPS-induced lung injury,which may represent a promising strategy to treat ALI.展开更多
Secreted protein acidic and rich in cysteine(SPARC)is a matricellular protein highly expressed in bone tissue that acts as achemoattractant factor promoting the arrival of prostate cancer(PCa)cells to the bone marrow....Secreted protein acidic and rich in cysteine(SPARC)is a matricellular protein highly expressed in bone tissue that acts as achemoattractant factor promoting the arrival of prostate cancer(PCa)cells to the bone marrow.However,the contribution of SPARCduring the early stages of tumor progression remains unclear.In this study,we show that SPARC is highly expressed in PCa tissueswith a higher Gleason score.Through stable knockdown and overexpression of SPARC in PC3 and LNCaP cells,respectively,here wedem on strate that en doge nous SPARC induces the epithelial-mesenchymal tran sition(EMT),decreasing E-cadheri n and cytokeratin18 and increasing N-cadheri n and vime ntin.Moreover,SPARC in duces the expression of EMT regulatory tran scription factors Snailfamily transcriptional repressor 1(Snail),Snail family transcriptional repressor 2(Slug),and zinc finger E-box binding homeobox 1(Zeb1).In addition,SPARC knockdown in PC3 cells decreases migration and invasion in vitro,without modifying cell proliferation.Our results indicate that SPARC might facilitate tumor progression by modifying the cellular phenotype in cancer cells.展开更多
Genetic association studies have implicated that cartilage intermediate layer protein 2 (CILP-2) confers the risk susceptibility for type 2 diabetes (T2DM). However, it is still unknown whether CILP-2 is involved in t...Genetic association studies have implicated that cartilage intermediate layer protein 2 (CILP-2) confers the risk susceptibility for type 2 diabetes (T2DM). However, it is still unknown whether CILP-2 is involved in the regulation of glucose homeostasis and insulin resistance (IR). In the current study, we initially observed that CILP-2 as a secreted protein was detected in both conditioned medium and lysates of cells transfected with an overexpressed vector. We then found that circulating CILP-2 levels had a progressive increase from normal to impaired glucose tolerance (a pre-diabetic status) and then to diabetes, which was correlated positively with waist-to-hip ratio, triglyceride, fasting blood glucose, 2-h blood glucose after glucose overload, HbA1c, fasting insulin, 2-h plasma insulin after glucose overload, and homeostasis model assessment of insulin resistance but negatively with HDL-C. CILP-2 expression was increased in the liver and muscle but decreased in adipose tissues of obese mice or T2DM patients. Furthermore, we demonstrated that CILP-2 circulating levels were affected by OGTT and Exenatide. CILP-2 overexpression resulted in impaired glucose tolerance and hepatic IR in vivo and increased PEPCK expression whereas suppressed phosphorylation of insulin receptor and Akt kinase in vitro. Based on these findings, we have identified a direct interaction between CILP-2 and PEPCK and suggested that CILP-2 plays an important role in the regulation of hepatic glucose production.展开更多
In paper[Chin.Phys.B 32070308(2023)],Xing et al.proposed a semi-quantum secret sharing protocol by using single particles.We study the security of the proposed protocol and find that it is not secure,that is,the three...In paper[Chin.Phys.B 32070308(2023)],Xing et al.proposed a semi-quantum secret sharing protocol by using single particles.We study the security of the proposed protocol and find that it is not secure,that is,the three dishonest agents,Bob,Charlie and Emily can collude to obtain Alice's secret without the help of David.展开更多
Cryptography is deemed to be the optimum strategy to secure the data privacy in which the data is encoded ahead of time before sharing it.Visual Secret Sharing(VSS)is an encryption method in which the secret message i...Cryptography is deemed to be the optimum strategy to secure the data privacy in which the data is encoded ahead of time before sharing it.Visual Secret Sharing(VSS)is an encryption method in which the secret message is split into at least two trivial images called’shares’to cover it.However,such message are always targeted by hackers or dishonest members who attempt to decrypt the message.This can be avoided by not uncovering the secret message without the universal share when it is presented and is typically taken care of,by the trusted party.Hence,in this paper,an optimal and secure double-layered secret image sharing scheme is proposed.The proposed share creation process contains two layers such as threshold-based secret sharing in the first layer and universal share based secret sharing in the second layer.In first layer,Genetic Algorithm(GA)is applied to find the optimal threshold value based on the randomness of the created shares.Then,in the second layer,a novel design of universal share-based secret share creation method is proposed.Finally,Opposition Whale Optimization Algorithm(OWOA)-based optimal key was generated for rectange block cipher to secure each share.This helped in producing high quality reconstruction images.The researcher achieved average experimental outcomes in terms of PSNR and MSE values equal to 55.154225 and 0.79365625 respectively.The average PSNRwas less(49.134475)and average MSE was high(1)in case of existing methods.展开更多
基金the grants from the National Natural Science Foundation of China(U1805232,31770156,and 32172365)the China Postdoctoral Science Foundation(2021M690637)。
文摘The fine-tuned expression dynamics of the effector genes are pivotal for the transition from vegetative growth to host colonization of pathogenic filamentous fungi.However,mechanisms underlying the dynamic regulation of these genes remain largely unknown.Here,through comparative transcriptome and chromatin immunoprecipitation sequencing(ChIP-seq)analyses of the methyltransferase PoKmt6 in rice blast fungus Pyricularia oryzae(syn.Magnaporthe oryzae),we found that PoKmt6-mediated H3K27me3 deposition was enriched mainly at fast-evolving regions and contributed to the silencing of a subset of secreted proteins(SP)and transposable element(TE)families during the vegetative growth of P.oryzae.Intriguingly,we observed that a group of SP genes,which were depleted of H3K27me3 modification,could also be silenced via the H3K27me3-mediated repression of the nearby TEs.In conclusion,our results indicate that H3K27me3 modification mediated by PoKmt6 regulates the expression of some SP genes in fast-evolving regions through the suppression of nearby TEs.
文摘Objective Obstructive sleep apnea (OSA) is closely related to obesity, insulin resistance and inflammation. Secreted frizzled-related protein 5 (SFRP5) is a recently discovered adipokine. It is involved in insulin resistance and inflammation in obesity. This study aimed at evaluating the association between SFRP5and sleeping characteristics as well as biochemical parameters of OSA patients.Methods This was a prospective case control study. Nondiabetic OSA patients and controls were consecutively recruited and divided into three groups: OSA group, apnea–hypopnea Index (AHI)≥5/h; healthy controls with normal body mass index (BMI); obese controls without OSA, and BMI > 24.0 kg/m2. All participants underwent polysomnography (PSG). Plasma SFRP5 was examined using enzyme-linked immunosorbent assay (ELISA). Blood biochemical examinations, including fasting blood glucose (FBG), lipid profile, hypersensitive Creactive protein (hsCRP), were performed early in the morning after PSG. Patients with severe OSA were treated with nasal continuous positive airway pressure (nCPAP), and plasma SFRP5 was repeatedly measured for comparison.Results Sixty-eight subjects were enrolled in the study, including 38 patients of OSA, whose medium AHI was 58.70 /h (36.63, 71.15), 20 obese controls, and 10 healthy controls. The plasma SFRP5 level of OSA patients was not significantly different from that of healthy controls or obese controls. In OSA patients, SFRP5 level correlated positively with triglyceride level (r=0.447, P=0.005) and negatively with LDL-cholesterol level and HDLcholesterol level (r=?0.472 and P=0.003; r=?0.478 and P=0.002; respectively). SFRP5 level was not found correlating with FBG, AHI, or any of nocturnal hypoxia parameters. After overnight nCPAP treatment, plasma SFRP5 levels of OSA patients did not change significantly (t=1.557, P = 0.148) compared to that of pretreatment.Conclusions In nondiabetic OSA patients, plasma SFRP5 is associated with the lipid profile. However,no correlation was observed between SFRP5 and FBG or sleep parameters. The SFRP5 level of OSA patients did not differ from that of non-OSA individuals in our study.
基金Basic Public Welfare Research Foundation of Zhejiang Province,China,No.GD21H290001and Traditional Chinese Medicine Science and Technology Project Foundation of Zhejiang Province,China,No.2020ZB072.
文摘BACKGROUND Sepsis is a major medical challenge.Magnolol is an active constituent of Houpu that improves tissue function and exerts strong anti-endotoxin and anti-inflammatory effects,but the mechanism by which it reduces intestinal inflammation in sepsis is yet unclear.AIM To assess the protective effect of magnolol on intestinal mucosal epithelial cells in sepsis and elucidate the underlying mechanisms.METHODS Enzyme-linked immunosorbent assay was used to measure tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),IL-6,and regulated on activation,normal T-cell expressed and secreted(RANTES)levels in serum and ileal tissue in animal studies.The histopathological changes of the ileal mucosa in different groups were observed under a microscope.Cell Counting Kit-8 and cell permeability assays were used to determine the concentration of drug-containing serum that did not affect the activity of Caco2 cells but inhibited lipopolysaccharide(LPS)-induced decrease in permeability.Immunofluorescence and Western blot assays were used to detect the levels of RANTES,inhibitor of nuclear factor kappa-B kinaseβ(IKKβ),phosphorylated IKKβ(p-IKKβ),inhibitor of nuclear factor kappa-B kinaseα(IκBα),p65,and p-p65 proteins in different groups in vitro.RESULTS In rats treated with LPS by intravenous tail injection in the presence or absence of magnolol,magnolol inhibited the expression of proinflammatory cytokines,IL-1β,IL-6,and TNF-αin a dose-dependent manner.In addition,magnolol suppressed the production of RANTES in LPS-stimulated sepsis rats.Moreover,in vitro studies suggested that magnolol inhibited the increase of p65 nucleation,thereby markedly downregulating the production of the phosphorylated form of IKKβin LPS-treated Caco2 cells.Specifically,magnolol inhibited the translocation of the transcription factor nuclear factor-kappa B(NF-κB)from the cytosol into the nucleus and down-regulated the expression level of the chemokine RANTES in LPS-stimulated Caco2 cells.CONCLUSION Magnolol down-regulates RANTES levels by inhibiting the LPS/NF-κB signaling pathways,thereby suppressing IL-1β,IL-6,and TNF-αexpression to alleviate the mucosal barrier dysfunction in sepsis.
基金This work is supported by the National Natural Science Foundation of China （30360061）Natural Science Foundation of Yunnan Province of China （1999C0008Z） National 863 Program of China （2003AA211020）.
文摘Computer based software such as the SignalP v3.0, TargetP v1.01, big-PI predictor and TMHMM v2.0 were combined topredict the signal peptides, and the signal peptide-dependent secreted proteins among the 6 700 ORFs in genome ofSaccharomyces cerevisiae. The results showed that 163 proteins were the secreted ones containing signal peptides, andthey were secreted via Sec pathway. Among the 163 predicted secreted proteins, the signal peptides of 47 secretedproteins included only the H-domain and C-domain, without N-domain, but the signal peptides of other 116 secretedproteins included all the three domains. There were differences in the constitution of signal peptides between the secretedproteins of S. cerevisiae and of Candida albicans, but the length and amino acids types of their signal peptides weresimilar in general. Few of the same signal peptides occurred in the secreted proteins of S. cerevisiae genome, and thehomology could be compared among the secreted proteins with the same signal peptides. The BLAST 2 SEQUENECESand CLUSTAL W were used to align the two protein sequences and multi-protein sequences, respectively. The alignmentresult indicated that homology of these sequences with the same signal peptide was very highly conservative in aminoacid of complete gene. The effect of the signal peptides in S. cerevisia on expression of foreign eukaryotic secretedproteins is discussed in this paper.
基金Supported by the Natural Science Foundation of Liaoning Province,No.201602817
文摘BACKGROUND Aberrant methylation in DNA regulatory regions could downregulate tumor suppressor genes without changing the sequences.However,our knowledge of secreted protein acidic and rich in cysteine(SPARC)and its aberrant methylation in gastric cancer(GC)is still inadequate.In the present research,we performed fundamental research to clarify the precise function of methylation on SPARC and its significance in GC.AIM To investigate promoter methylation and the effects of the SPARC gene in GC cells and tissues and to evaluate its clinical significance.METHODS Plasmids that overexpressed the SPARC gene were transfected into human GC BGC-823 cells;non-transfected cells were used as a control group(NC group).Quantitative real-time polymerase chain reaction and western blotting(WB)were then used to detect the expression of SPARC.Methylation-specific polymerase chain reaction was executed to analyze the gene promoter methylation status.Cell viability was measured by the cell counting kit-8 assay.The migration and invasion ability of cells were detected by scratch assays and transwell chamber assays,respectively.Cell cycle events and apoptosis were observed with a flow cytometer.RESULTS The expression of SPARC mRNA in GC tissues and cells was significantly lower and showed differing degrees of hypermethylation,respectively,than that in normal adjacent tissues and control cells.Treatment with 5-Aza-2’-deoxycytidine(5-Aza-Cdr)was able to restore the expression of SPARC and reverse promoter hypermethylation.Overexpression of the SPARC gene significantly inhibited proliferation,migration,and invasion of GC cells,while also causing cell cycle arrest and apoptosis;the NC group exhibited the opposite effects.CONCLUSION This study demonstrated that SPARC could function as a tumor suppressor and might be silenced by promoter hypermethylation.Furthermore,in GC cells,SPARC inhibited migration,invasion,and proliferation,caused cell cycle arrest at the G0/G1 phase,and promoted apoptosis.
基金the National Natural Science Foundation of China, No. 30270491 the Natural Science Foundation of Guangdong Province, No. 04020422 Science and Technology Plan Program of Guangdong Province, No. 2003A3020304
文摘BACKGROUND: It has been proved by many experimental studies from the aspects of morphology and immunocytochemistry in recent years that bone marrow stromal cells (BMSCs) can in vitro induce and differentiate into the cells possessing the properties of nerve cells. But the functions of BMSCs-derived neural stem cells(NSCs) and the differentiated neuron-like cells are still unclear. OBJECTIVE: To observe whether bone marrow-derived NSCs can secrete norepinephrine (NE) under the condition of in vitro culture, induce and differentiation, and analyze the biochemical properties of BMSCs-derived NSCs. DESIGN: A non-randomized and controlled experimental observation. SETTING: Institute of Neuromedicine of Chinese PLA, Zhujiang Hospital, Southern Medical University. MATERIALS: This experiment was carried out in the Institute of Neuromedicine of Chinese PLA, Zhujiang Hospital, Southern Medical University. The bone marrow used in the experiment was collected from 1.5- month-old healthy New Zealand white rabbits. METHODS: This experiment was carried out in the Institute of Neuromedicine of Chinese PLA, Zhujiang Hospital, Southern Medical University. The bone marrow used in the experiment was collected from 1.5 month-old healthy New Zealand white rabbits. BMSCs of rabbits were isolated and performed in vitro culture, induce and differentiation with culture medium of NSCs and differentiation-inducing factor, then identified with immunocytochemical method. Experimental grouping: ①Negative control group: L-02 hepatic cell and RPMI1640 culture medium were used. ② Background culture group: Only culture medium of NSCs as culture solution was added into BMSCs to perform culture, and 0.1 volume fraction of imported fetal bovine serum was supplemented 72 hours later. ③Differentiation inducing factor group: After culture for 72 hours, retinoic acid and glial cell line-derived neurotrophic factors were added in the culture medium of BMSCs and NSCs as corresponding inducing factors. The level of NE in each group was detected on the day of culture and 5, 7, 14 and 20 days after culture with high performance liquid chromatography(HPLC). The procedure was conducted 3 times in each group.Standard working curve was made according to the corresponding relationship of NE concentration and peak area. The concentration of NE every 1×107 cells was calculated according to standard curve and cell counting. MAIN OUTCOME MEASURES: The level of NE of cultured cells was detected with HPLC; immunocytochemistrical identification of Nestin and neuron specific nuclear protein was performed. RESULTS: ① On the 14th day after cell culture, BMSCs turned into magnus and round cells which presented Nestin-positive antigen, then changed into neuron-like cells with long processus and presented neuron specific nuclear protein -positive antigen at the 20th day following culture. ② The ratio of NE concentration and peak area has good linear relationship, and regression equation was Y=1.168 36+0.000 272 8X,r=0.998 4.Coefficient variation (CV) was < 5% and the recovery rate was 92.39%(Y referred to concentration and X was peak area).③NE was well detached within 10 minutes under the condition of this experiment. ④ NE was detected in NSCs and their culture mediums, which were cultured for 7, 14 and 20 days respectively, but no NE in BMSCs, NSCs-free culture medium and L-02 hepatic cell which were as negative control under the HPLC examination. Analysis of variance showed that the level of NE gradually increased following the elongation of culture time (P < 0.01). No significant difference in the level of NE existed at the same time between differentiation inducing factor group and basic culture group(P > 0.05). CONCLUSION: BMSCs of rabbits can proliferate in vitro and express Nestin antigen; They can differentiate into neuron-like cells, express specific neucleoprotein of mature neurons, synthesize and secrete NE as a kind of neurotransmitter.
文摘Protein secretion plays an important role in bacterial lifestyles. In Gram-negative bacteria, a wide range of proteins are secreted to modulate the interactions of bacteria with their environments and other bacteria via various secretion systems. These proteins are essential for the virulence of bacteria, so it is crucial to study them for the pathogenesis of diseases and the development of drugs. Using amino acid composition (AAC), position-specific scoring matrix (PSSM) and N-terminal signal peptides, two different substitution models are firstly constructed to transform protein sequences into numerical vectors. Then, based on support vector machine (SVM) and the “one to one”?algorithm, a hybrid multi-classifier named SecretP v.2.2 is proposed to rapidly and accurately?distinguish different types of Gram-negative?bacterial secreted proteins. When performed on the same test set for a comparison with other methods, SecretP v.2.2 gets the highest total sensitivity of 93.60%. A public independent dataset is used to further test the power of SecretP v.2.2 for predicting NCSPs, it also yields satisfactory results.
文摘In order to analyze the in vivo expression of Candida albicans secreted aspartyl pro- teinases (SAP) in human vaginal infection, the vaginal secretion from 29 human subjects was col- lected by vaginal swab, and the expression of SAP1–SAP6 was detected by reverse-transcriptase po- lymerase chain reaction using specific primer sets. It was found that Sap2 and Sap5 were the most common genes expressed during infection; Sap3 and Sap4 were detected in all subjects and all 6 SAP genes were simultaneously expressed in some patients with vaginal candidiasis. It was suggested that the SAP family is expressed by Candida albicans during infection in human and that Candida albi- cans infection is associated with the differential expression of individual SAP genes which may be involved in the pathogenesis of vaginal candidiasis.
基金Supported by the National Natural Science Foundation of China,No.81672041the National Major Science and Technology Special Project for Infectious Diseases of China,No.2012ZX10004503-012
文摘BACKGROUND Previously,we have successfully constructed replication-competent hepatitis B virus(HBV)vectors by uncoupling the P open reading frame(ORF)from the preC/C ORF to carefully design the transgene insertion site to overcome the compact organization of the HBV genome and maintain HBV replication competence.Consequently,the replication-competent HBV vectors carrying foreign genes,including pCH-BsdR,carrying blasticidin resistance gene(399 bp),and pCH-hrGFP,carrying humanized renilla green fluorescent protein gene(720 bp),were successfully obtained.However,the replication efficiency of the former is higher but it is tedious to use,while that of the latter is poor and cannot be quantified.Hence,we need to search for a new reporter gene that is convenient and quantifiable for further research.AIM To establish a helpful tool for intracellular HBV replication and anti-viral drugs screening studies.METHODS We utilized the replication-competent HBV viral vectors constructed by our laboratory,combined with the secreted luciferase reporter gene,to construct replication-competent HBV vectors expressing the reporter gene secretory Nanoluc Luciferase(SecNluc).HepG2.TA2-7 cells were transfected with this vector to obtain cell lines with stably secreted HBV particles carrying secNluc reporter gene.RESULTS The replication-competent HBV vector carrying the SecNluc reporter gene pCHsNLuc could produce all major viral RNAs and a full set of envelope proteins and achieve high-level secreted luciferase expression.HBV replication intermediates could be produced from this vector.Via transfection with pTRE-sNLuc and selection by hygromycin,we obtained isolated cell clones,named HBV-NLuc-35 cells,which could secrete secNLuc recombinant viruses,and were sensitive to existing anti-HBV drugs.Using differentiated HepaRG cells,it was verified that recombinant HBV possessed infectivity.CONCLUSION Our research demonstrated that a replication-competent HBV vector carrying a secreted luciferase transgene possesses replication and expression ability,and the established HBV replication and expression cell lines could stably secrete viral particles carrying secNluc reporter gene.More importantly,the cell line and the secreted recombinant viral particles could be used to trace HBV replication or infection.
文摘BACKGROUND Secreted protein acidic and rich in cysteine(SPARC)is an extracellular matrixassociated protein.Studies have revealed that SPARC is involved in the cell interaction and function including proliferation,differentiation,and apoptosis.However,the role of SPARC in cancer is controversial,as it was reported as the promoter or suppressor in different cancers.Further,the role of SPARC in lymphoma is unclear.AIM To identify the expression and significance of SPARC in lymphoma,especially in diffuse large B-cell lymphoma(DLBCL).METHODS The expression analysis of SPARC in different cancers was evaluated with Oncomine.The Brune,Eckerle,Piccaluga,Basso,Compagno,Alizadeh,and Rosenwald datasets were included to evaluate the mRNA expression of SPARC in lymphoma.The Cancer Genome Atlas(TCGA)-DLBCL was used to analyze the diagnostic value of SPARC in DLBCL.The Compagno and Brune DLBCL datasets were used for validation.Then,the diagnostic value was evaluated with the receiver operating characteristic(ROC)curve.The Kaplan-Meier plot was conducted with TCGA-DLBCL,and the ROC analysis was performed based on the survival time.Further,the overall survival analysis based on the level of SPARC expression was performed with the GSE4475 and E-TABM-346.The Gene Set Enrichment Analyses(GSEA)was performed to make the underlying mechanism-regulatory networks.RESULTS The pan-cancer analysis of SPARC showed that SPARC was highly expressed in the brain and central nervous system,breast,colon,esophagus,stomach,head and neck,pancreas,and sarcoma,especially in lymphoma.The overexpression of SPARC in lymphoma,especially DLBCL,was confirmed in several datasets.The ROC analysis revealed that SPARC was a valuable diagnostic biomarker.More importantly,compared with DLBCL patients with low SPARC expression,those with higher SPARC expression represented a higher overall survival rate.The ROC analysis showed that SPARC was a favorable prognostic biomarker for DLBCL.Results of the GSEA confirmed that the high expression of SPARC was closely associated with focal adhesion,extracellular matrix receptor interaction,and leukocyte transendothelial migration,which suggested that SPARC may be involved in the regulation of epithelial-mesenchymal transition,KRAS,and myogenesis in DLBCL.CONCLUSION SPARC was highly expressed in DLBCL,and the overexpression of SPARC showed sound diagnostic value.More interestingly,the overexpression of SPARC might be a favorable prognostic biomarker for DLBCL,suggesting that SPARC might be an inducible factor in the development of DLBCL,and inducible SPARC was negative in some oncogenic pathways.All the evidence suggested that inducible SPARC might be a good diagnostic and prognostic biomarker for DLBCL.
基金Supported by National Basic Research Program(2012CB722901)Academic Award for Up-and-coming Doctoral Candidates of Yunnan ProvinceYunnan Agricultural University Innovation Foundation for Postgraduate
文摘The structure similarity of secreted proteins in rice blast fungus Magnaporthe oryzae and its host Oryza sativa was analyzed. One thousand two hundred and forty one proteins were predicted as secreted proteins using four algorithms based on 11 074 proteins in genome of M. oryzae. One hundred and forty six secreted proteins( 11. 8% of M. oryzae secretome) were aligned with 116 rice proteins( 0. 21% of 56 278 rice proteins) using BLAST search on rice genome. One hundred sixteen rice similar proteins participated in rice cell wall modification( cell wall associated enzymes) and signal transduction( proteases). These results imply that both cell wall involved proteins and signal transduction are probably hijacks pathway between host pants and pathogenic fungi. Because these proteins are highly conserved among fungi and plants,the express patterns of these protein coding genes during the interaction process are valuable to study in detail.
文摘Corynebacterium pseudotuberculosis is an infectious agent that occurs in small ruminants causing caseous lymphadenitis, and more rarely in humans causing lymphadenitis and pneumonia. The breeding small ruminants have great economic importance in Brazil. Rural farm workers and veterinary students who acquired this disease suffered from weakening symptoms for weeks, and the identification of the etiological agent was time-consuming and complex. Due to the low prevalence of case records, there is probably no available commercial diagnostic kit for C. pseudotuberculosis infection in humans. This study aimed to describe human seroreactivity to secreted antigens from C. pseudotuberculosis. Reactivity of serum from farm workers (n = 14), individuals who work with the bacillus at laboratory (n = 8) or individuals without contact (n = 25) was tested with secreted proteins from PAT10 strain of C. pseudotuberculosis by Western blotting. Samples of all (100%) farm workers showed reactivity to 31 kDa, 71 kDa and 164 kDa proteins, while laboratory workers showed 87.5%, 62.5 % and 37.5%, and no-contact 20%, 0% and 16%, respectively. All sera recognized the 275 kDa protein. Our data suggest that C. pseudotuberculosis secreted proteins are antigenic in humans and the recognition profiles allowed the identification of individuals with and without prior contact with this bacillus. This is the first paper which describes human reactivity to C. pseudotuberculosis in serum samples of workers in Brazil.
文摘Phosphorus is one of the most important nutrients required for plant growth and development. While substantial amounts of total phosphorus are present in many soil types, plants are unable to utilize some organic phosphorus sources. The main goal of this study was to characterize the spectrum of secreted plant proteins, organic acids and other metabolites that can potentially contribute to utilization of various phosphorus compounds. Our data indicate that the composition of extracellular proteins secreted by plant roots varies depending on the specific source of P in the growth medium. Furthermore, some root-secreted metabolites, such as citrate, appear to be specific to a subset of ecotypes, while tartrate, succinate and oxalate are secreted by a number of A. thaliana ecotypes. We observed secretion of phenolic compounds, such as tannins, and deoxycytidine derivatives. Taken together, while no single secreted polypeptide, organic acid or secondary metabolite can be pinpointed as specific to plant growth in particular phosphorus conditions, our data indicate that A. thaliana ecotypes differ in their physiological responses to the source of phosphorus in the growth medium. Overall, these results suggest that physiological changes in plant responses to nutrient limitation are modulated by interactions between soil phosphorus source and the specific genotype of Arabidopsis plants.
文摘Background:Cancer-associated fibroblasts(CAFs)play an important role in the induction of chemo-resistance.This study aimed to clarify the mechanism underlying CAF-mediated resistance to two tyrosine kinase inhibitors(TKIs),sorafenib and lenvatinib,and to identify a novel therapeutic target for overcoming TKI resistance in hepatocellular carcinoma(HCC).Methods:We performed a systematic integrative analysis of publicly available gene expression datasets and whole-transcriptome sequencing data from 9 pairs of CAFs and para-cancer fibroblasts isolated from human HCC and para-tumor tissues,respectively,to identify key molecules that might induce resistance to TKIs.We then performed in vitro and in vivo experiments to validate selected targets and related mechanisms.The associations of plasma secreted phosphoprotein 1(SPP1)expression levels before sorafenib/lenvatinib treatment with progression-free survival(PFS)and overall survival(OS)of 54 patients with advanced HCC were evaluated using Kaplan-Meier and Cox regression analysis.Results:Bioinformatic analysis identified CAF-derived SPP1 as a candidate molecule driving TKI resistance.SPP1 inhibitors reversed CAF-induced TKI resistance in vitro and in vivo.CAF-derived SPP1 activated rapidly accelerated fibrosarcoma(RAF)/mitogen-activated protein kinase(MAPK)and phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)/mammalian target of rapamycin(mTOR)through the integrin-protein kinase C-alpha(PKCα)signaling pathway and promoted epithelial-to-mesenchymal transition(EMT).A high plasma SPP1 level before TKI treatment was identified as an independent predictor of poor PFS(P=0.026)and OS(P=0.047)in patients with advanced HCC after TKI treatment.Conclusions:CAF-derived SPP1 enhances TKI resistance in HCC via bypass activation of oncogenic signals and EMT promotion.Its inhibition represents a promising therapeutic strategy against TKI resistance inHCC.Moreover,plasma SPP1 level before TKI treatment represents a potential biomarker for treatment response prediction.
基金supported by Shandong Natural Science Foundation[grant number ZR2021MH339].
文摘Background and aims:Secreted phosphoprotein 1(SPP1)functions in several physiological processes.The role of SPP1 expression in the prognosis and tumor immunity of hepatocellular carcinoma(HCC)is unknown.The aim of this study was to investigate the expression pattern of SPP1 in HCC and its correlation with prognosis and tumor immunity.Methods:Clinical and gene expression data of The Cancer Genome Atlas-liver hepatocellular carcinoma(LIHC)cohort and 11 other HCC datasets were collected.The Kaplan–Meier method and Cox regression analysis were used to analyze the prognostic value of SPP1.The DESeq2 package in R was used to analyze SPP1-related genes.Gene Ontology analysis and gene set enrichment analysis were used to determine the biological function of SPP1 in HCC.The single sample Gene Set Enrichment Analysis(ssGSEA)method was used to analyze the immune infiltrates of HCC.Illumina human methylation 450 data and level 3 HTSeq-FPKM data from The Cancer Genome Atlas-LIHC were used to analyze the effects of DNA methylation level on SPP1 expression.Results:SPP1 was overexpressed in HCC and correlated with T stage,histological grade,adjacent hepatic tissue inflammation,and vascular invasion in HCC.The analysis of survival rates indicated that high SPP1 levels were associated with poor overall survival in HCC.Functional analysis showed that SPP1 is related to tumor immunity,especially macrophage infiltration.Aberrant demethylation of the promoter region is one of the mechanisms underlying the increase of SPP1 in HCC.Conclusion:Our results indicate that SPP1 is an independent prognostic factor for HCC and is correlated with the clinical features and macrophage infiltration in HCC.
基金the National Natural Science Foundation(81773982,82003937)Youth Academic leaders of the Qinglan Project in Jiangsu province for financial support。
文摘In the face of increasingly serious environmental pollution,the health of human lung tissues is also facing serious threats.Mogroside IIE(M2E)is the main metabolite of sweetening agents mogrosides from the anti-tussive Chinese herbal Siraitia grosvenori.The study elucidated the anti-inflammatory action and molecular mechanism of M2E against acute lung injury(ALI).A lipopolysaccharide(LPS)-induced ALI model was established in mice and MH-S cells were employed to explore the protective mechanism of M2E through the western blotting,co-immunoprecipitation,and quantitative real time-PCR analysis.The results indicated that M2E alleviated LPS-induced lung injury through restraining the activation of secreted phospholipase A2 type IIA(Pla2g2a)-epidermal growth factor receptor(EGFR).The interaction of Pla2g2a and EGFR was identified by co-immunoprecipitation.In addition,M2E protected ALI induced with LPS against inflammatory and damage which were significantly dependent upon the downregulation of AKT and m TOR via the inhibition of Pla2g2a-EGFR.Pla2g2a may represent a potential target for M2E in the improvement of LPS-induced lung injury,which may represent a promising strategy to treat ALI.
文摘Secreted protein acidic and rich in cysteine(SPARC)is a matricellular protein highly expressed in bone tissue that acts as achemoattractant factor promoting the arrival of prostate cancer(PCa)cells to the bone marrow.However,the contribution of SPARCduring the early stages of tumor progression remains unclear.In this study,we show that SPARC is highly expressed in PCa tissueswith a higher Gleason score.Through stable knockdown and overexpression of SPARC in PC3 and LNCaP cells,respectively,here wedem on strate that en doge nous SPARC induces the epithelial-mesenchymal tran sition(EMT),decreasing E-cadheri n and cytokeratin18 and increasing N-cadheri n and vime ntin.Moreover,SPARC in duces the expression of EMT regulatory tran scription factors Snailfamily transcriptional repressor 1(Snail),Snail family transcriptional repressor 2(Slug),and zinc finger E-box binding homeobox 1(Zeb1).In addition,SPARC knockdown in PC3 cells decreases migration and invasion in vitro,without modifying cell proliferation.Our results indicate that SPARC might facilitate tumor progression by modifying the cellular phenotype in cancer cells.
基金the National Natural Science Foundation of China(81670755,81601214,81873658,81570752,81100567,and 81800755)Natural Science Foundation Key Project of Chongqing(cstc 2015jcyjA10084)the Science and Tech no logy Key Program of Health Bureau of Chongqing(2015ZDXM038).
文摘Genetic association studies have implicated that cartilage intermediate layer protein 2 (CILP-2) confers the risk susceptibility for type 2 diabetes (T2DM). However, it is still unknown whether CILP-2 is involved in the regulation of glucose homeostasis and insulin resistance (IR). In the current study, we initially observed that CILP-2 as a secreted protein was detected in both conditioned medium and lysates of cells transfected with an overexpressed vector. We then found that circulating CILP-2 levels had a progressive increase from normal to impaired glucose tolerance (a pre-diabetic status) and then to diabetes, which was correlated positively with waist-to-hip ratio, triglyceride, fasting blood glucose, 2-h blood glucose after glucose overload, HbA1c, fasting insulin, 2-h plasma insulin after glucose overload, and homeostasis model assessment of insulin resistance but negatively with HDL-C. CILP-2 expression was increased in the liver and muscle but decreased in adipose tissues of obese mice or T2DM patients. Furthermore, we demonstrated that CILP-2 circulating levels were affected by OGTT and Exenatide. CILP-2 overexpression resulted in impaired glucose tolerance and hepatic IR in vivo and increased PEPCK expression whereas suppressed phosphorylation of insulin receptor and Akt kinase in vitro. Based on these findings, we have identified a direct interaction between CILP-2 and PEPCK and suggested that CILP-2 plays an important role in the regulation of hepatic glucose production.
基金Project supported by the Offline Course Program of“Experiment of College Physics”in the 2022-year Anhui Provincial Quality Engineering Program (Grant No.2022xxkc134)the Program for Academic Leader Reserve Candidates in Tongling University (Grant Nos.2020tlxyxs43 and 2014tlxyxs30)+1 种基金the Talent Scientific Research Foundation of Tongling University (Grant No.2015tlxyrc01)the 2014 year Program for Excellent Youth Talents in University of Anhui Province。
文摘In paper[Chin.Phys.B 32070308(2023)],Xing et al.proposed a semi-quantum secret sharing protocol by using single particles.We study the security of the proposed protocol and find that it is not secure,that is,the three dishonest agents,Bob,Charlie and Emily can collude to obtain Alice's secret without the help of David.
基金supported by RUSA PHASE 2.0,Alagappa University,Karaikudi,India。
文摘Cryptography is deemed to be the optimum strategy to secure the data privacy in which the data is encoded ahead of time before sharing it.Visual Secret Sharing(VSS)is an encryption method in which the secret message is split into at least two trivial images called’shares’to cover it.However,such message are always targeted by hackers or dishonest members who attempt to decrypt the message.This can be avoided by not uncovering the secret message without the universal share when it is presented and is typically taken care of,by the trusted party.Hence,in this paper,an optimal and secure double-layered secret image sharing scheme is proposed.The proposed share creation process contains two layers such as threshold-based secret sharing in the first layer and universal share based secret sharing in the second layer.In first layer,Genetic Algorithm(GA)is applied to find the optimal threshold value based on the randomness of the created shares.Then,in the second layer,a novel design of universal share-based secret share creation method is proposed.Finally,Opposition Whale Optimization Algorithm(OWOA)-based optimal key was generated for rectange block cipher to secure each share.This helped in producing high quality reconstruction images.The researcher achieved average experimental outcomes in terms of PSNR and MSE values equal to 55.154225 and 0.79365625 respectively.The average PSNRwas less(49.134475)and average MSE was high(1)in case of existing methods.
