Sophora japonica are used as a medicinal herb in China, Japan and Korea to treat bleeding hemorrhoids and hematemesis. This article presents an overview of the effects of Sophora japonica on cerebral infarction based on literature searched from Medline, PubMed, Cochrane Library and the China National Knowledge Infrastructure (CNKI). Sophora japonica contains both anti-hemorrhagic and anti-hemostatic substances. Sophora japonica reduces cerebral infarction partly as a result of its anti-oxidative and anti-inflammatory activities. Previous studies found that Sophora japonica reduced the size of cerebral infarction and neurological deficits and reduced microglial activation, interleukin-1β release and number of apoptotic cells in ischemia-reperfusion injured Sprague-Dawley rats. Further study is required to determine the relationship between Sophora japonica-mediated reduction in cerebral infarction size and the effects of Sophora japonica on platelet aggregation and cardiovascular function.The dried flowers and buds of  Sophora japonica (Huaihua) are a medicinal herb used in China, Japan and Korea to treat hemorrhoids and hematemesis [Sophora japonica include flavones, tetraglycosides, isoflavones, isoflavone tetraglycosides, triterpene glycosides, phospholipids, alkaloids, amino acids and polysaccharides [Sophora japonica contains five main flavonoids of rutin, quercetin, isorhamnetin, isorhamnetin, genistein and kaempferol [Dried flowers and buds of atemesis . Main cocharides . Moreoveempferol .Sophora japonica, in particular its effects and mechanisms in reducing cerebral infarction (Table Sophora japonica (OR quercetin OR rutin) AND cerebral infarction as the English keywords and Huaihua as the Chinese one. The initial search generated 173 articles from the English language databases and 141 articles from the Chinese one.This article aims to provide an overview of the pharmacology of on Table . To thisGuizhifuling Wan  reduced superoxide anion levels during 2 hours of reperfusion after ischemia, cerebral infarction size and neurological deficit score in rats with MCAo [Ginkgo biloba L. leaves which contain quercetin and rutin which have free radical scavenging activity [Ginkgo biloba L. leaves (Egb 761) reduced the size of cerebral infarction and improved neurological behavior in rats with permanent and transient MCAo [et al. found that the flavonoid components of Sophora japonica scavenged superoxide anion and 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radicals [Sophora japonica, Wang et al. found that the polysaccharides of Sophora japonica scavenged hydroxyl and superoxide anion free radicals [t-BuOOH-induced reactive oxygen species [et al. found that rutin in iron-rutin complex inhibited the conversion of superoxide anion to hydroxyl radicals in normal and iron-overloaded rats [et al. showed that the anti-oxidant irisolidone prevented lipid peroxidation, DNA damage and reduced apoptosis induced by hydrogen peroxide in Chinese hamster lung fibroblasts [Our previous studies found that (1) levels of superoxide anion in arterial blood increased at the start of 2 hours of reperfusion after ischemia in rats with middle cerebral artery occlusion (MCAo) ; (2) levith MCAo -5. Free activity . Extractaves Egb 61 reduceradicals . Simple radicals . Both qu species . Afanas'ded rats . Kang etroblasts .Gingko biloba extracts played a neuroprotective role [Generation of reactive oxygen species (ROS) including nitrogen and oxygen free radicals played a critical role in brain damage during the reperfusion period after ischemia ,14. Braiive role ,17. Bothive role . Rutin rive role .Guizhifuling Wan and paeonol inhibited microglial activation and IL-1β release and reduced cerebral infarction size in rats with MCAo [Sophora japonica inhibited microglial activation, interleukin-1β (IL-1β) release and apoptosis in rats with transient MCAo, suggesting that Sophora japonica reduced inflammation and prevented neuronal death by inhibiting microglial release of IL-1β, a pro-inflammatory cytokine [et al. reported that dietary rutin supplementation 10 or 20 days after trimethyltin (TMT) administration in rats reduced IL-1β mRNA expression in the microglia of the hippocampus, suggesting that rutin was neuro-protective against TMT-induced neuronal damage [Microglia was activated by various types of brain damage such as ischemic and inflammatory damage . This acith MCAo ,5. Sophocytokine . Koda etl damage .Sophora japonica were stronger inhibitors of arachidonic acid- and thromboxane A2-induced platelet aggregation in platelet-rich and platelet-poor plasma in rats [Sophora japonica inhibited free calcium accumulation within platelets thereby preventing platelet aggregation [Kim and Yun-Chol reported that, compared with acetylsalicylic acid, biochanin A, irisolidone, genistein and tectoridin of regation . Antiplaregation . Plateleregation .Sophora japonica decoction into the jugular veins of rabbits decreased cardiac muscle contractility and reduced heart rate, suggesting that Sophora japonica reduced the consumption of oxygen to protect cardiac function [Sophora japonica) was due to stabilization of capillary integrity [Sophora japonica extracts  for five days reduced capillary perameability, bleeding time and coagulation time in mice and reduced prothrombin time in rats, thereby demonstrating the hemostatic effect of Sophora japonica [Sophora japonica was also anti-hemostatic, as a result from increased capillary permeability and reduced platelet aggregation [Administration of function ,31. The ntegrity . Oral adjaponica ,33. IsorSophora japonica contains both anti-hemorrhagic and anti-hemostatic substances. Sophora japonica reduces cerebral infarction partly as a result of its anti-oxidative and anti-inflammatory activities. Further study is required to determine the relationship between Sophora japonica-mediated reduction in cerebral infarction size and its effects on platelet aggregation and cardiovascular function.MCAO: middle cerebral artery occlusion; EGB 761: extract of Ginkgo biloba L. leaves; DPPH: 1,1-diphenyl-2-picrylhydrazyl; T-BUOOH: tert-butyl hydroperoxide; IL-1β: interleukin-1β; TMT: trimethyltin; MRNA: messenger ribonucleic acid; ROS: reactive oxygen species; DNA: deoxyribonucleic acidThe authors declare that they have no competing interests.CLH designed the study and revised the manuscript. HNC conducted the literature search and drafted the manuscript. Both authors read and approved the final version of the manuscript.


Anopheles gambiae is a major vector of human malaria. Increasing evidence indicates that blood cells (hemocytes) comprise an essential arm of the mosquito innate immune response against both bacteria and malaria parasites. To further characterize the role of hemocytes in mosquito immunity, we undertook the first genome-wide transcriptomic analyses of adult female An. gambiae hemocytes following infection by two species of bacteria and a malaria parasite.The mosquito An. gambiae genome-wide microarrays. While 279 transcripts were significantly enriched in hemocytes relative to whole adult female mosquitoes, 959 transcripts exhibited immune challenge-related regulation. The global transcriptomic responses of hemocytes to challenge with different species of bacteria and/or different stages of malaria parasite infection revealed discrete, minimally overlapping, pathogen-specific signatures of infection-responsive gene expression; 105 of these represented putative immunity-related genes including anti-Plasmodium factors. Of particular interest was the specific co-regulation of various members of the Imd and JNK immune signaling pathways during malaria parasite invasion of the mosquito midgut epithelium.We identified 4047 genes expressed in hemocytes, using Our genome-wide transcriptomic analysis of adult mosquito hemocytes reveals pathogen-specific signatures of gene regulation and identifies several novel candidate genes for future functional studies. Insect blood cells (hemocytes) play a central role in mediating innate immune responses -4. HemocDrosophila melanogaster and Lepidoptera ) ) -45. We a [APL2])  that areic cells . As manyic cells ,48, and ic cells ,50. All asmodium ,51. ThirIn addition to canonical immunity-related genes, we detected several gene transcripts in hemocytes belonging to the cytoskeletal/structural functional class that have conserved roles in phagocytosis ,53. ThesAedes aegypti and Armigeres subalbatus . Onl. Onl3 heP. berghei-infected and control mosquitoes were prepared and collected as described above. A minimum of 30 individuals were bled and their hemocytes pooled to obtain at least 500 ng of total RNA per replicate. However, for the day 19 post-blood feeding samples, more than 100 mosquitoes were bled per sample due to the reduced number of hemocytes present per individual (see Results). Total RNAs were then isolated from hemocytes using the RNAEasy kit (Qiagen) as outlined by the manufacturer. For each treatment and time point, three independent samples were prepared for use in subsequent microarray and QRT-PCR analyses.Hemocyte samples from bacteria-infected, 300–400 ng of total RNA was used to synthesize Cy-3 or Cy-5 fluorochrome-labeled cRNA probes for each hemocyte or whole adult female sample using Agilent's Low RNA Input Linear Amplification Kit  according to the manufacturer's instructions. Two-color microarray hybridizations were performed using Agilent's In situ Hybridization Kit Plus (Cat. No. 5184-3568) and custom-made 60-mer oligonucleotide microarrays purchased from Agilent as previously reported . Arrays P. berghei challenged and 24 hr non-infected blood fed samples. Total RNA samples were reversed transcribed using dT20 primers and Superscript III . Real-time quantitative PCR assays were performed using QuantiTect SYBR Green PCR Kit  and ABI Detection System ABI Prism 7000 . The ribosomal protein S7 gene was used for normalization of cDNA templates, the specificity of the PCR reactions was confirmed by melting curves analysis. Primer sequences used for microarray validation have been previously published [This microarray gene expression platform has been previously validated  and we cublished . The expAMP: antimicrobial peptide; AP-1: adaptor protein complex 1; CEC: cecropin; cfu: colony forming unit; DEF: defensin; FBN: fibrinogen-domain-containing immunolectin; GALE: galectin; GAM: gambicin; GNBP: Gram-negative binding protein; HAT: histone acetyltransferase; HDAC: histone deacetylase; LPS: lipopolysaccharide; LYS: lysozyme; PGRP: peptidoglycan-recognition protein; PO: phenoloxidase; PPO: prophenoloxidase; PPR: pattern recognition receptor; RTQ-PCR: real-time quantitative polymerase chain reaction; TEP: thio-ester containing protein.LB carried out microarray assays and analyses, and participated in writing the manuscript. AR carried out hemocyte assays. EW carried out microarray assays and analyses. MS conceived of the study, and participated in its design and coordination and participated in writing the manuscript. GD conceived of the study, and participated in its design and coordination, the analysis of data and writing the manuscript. All authors read and approved the final manuscript.Table S1. Log2 transformed expression ratio microarray data for the various experimental comparisons. Significantly up- and down-regulated genes are indicated with either "Up" or "Down" in separate columns.Click here for fileAdditional text files S1 – S9. Additional texts that are complementary to their respective main text sections.Click here for file


Hepatocyte growth factor (HGF) is a multifunctional growth factor which has pleiotrophic biological effects on epithelial cells such as proliferation, motogenesis, invasiveness and morphogenesis. Peritoneal dissemination is critical for the progression of ovarian cancer, and our study revealed that HGF induces migration and invasion of ovarian cancer cells. We also demonstrated that HGF stimulates autophosphorylation of its receptor, followed by activation of the Ras-MAP (mitogen-activated peptide) kinase cascade. Moreover, infection of ovarian cancer cells with Ras dominant-negative adenovirus reduced the HGF-induced motogenic and invasive activities. Additionally, both MEK and PI3-kinase pathways downstream of Ras were involved in HGF-stimulated ovarian cancer cell invasiveness. © 2000 Cancer Research Campaign


Hormonally insensitive prostate cancer is a relatively slow-growing, but usually fatal, disease with no long-term treatment options. Transformation of normal prostate cells to a malignant phenotype often involves corruption of the apoptotic machineries. Bcl-2 protein is one of the key inhibitors of apoptosis and is often unregulated in advanced prostate cancer. The prostate cancer cell line DU-145 was used as a model of a hormonally insensitive, advanced prostate cancer. Cell growth in liquid culture was significantly inhibited by antisense Bcl-2 oligonucleotides compared with control sense oligonucleotides; inhibition by these oligonucleotides was significantly enhanced on combination with the synthetic retinoid N-(2-hydroxyphenyl)all-trans-retinamide (2-HPR). Interestingly, growth inhibition occurred in the absence of apoptosis as measured using two assay techniques. We hypothesize that in these recalcitrant cells the apoptotic pathway is compromised at several levels, and Bcl-2 may play another role in promoting cell growth. The use of Bcl-2 antisense oligonucleotides plus 2-HPR may provide a novel approach to therapy of hormone-resistant prostate cancer.


Reactive oxygen species (ROS) are known mediators of cellular damage in multiple diseases including diabetic complications. Despite its importance, no comprehensive database is currently available for the genes associated with ROS.We present ROS- and diabetes-related targets (genes/proteins) collected from the biomedical literature through a text mining technology. A web-based literature mining tool, SciMiner, was applied to 1,154 biomedical papers indexed with diabetes and ROS by PubMed to identify relevant targets. Over-represented targets in the ROS-diabetes literature were obtained through comparisons against randomly selected literature. The expression levels of nine genes, selected from the top ranked ROS-diabetes set, were measured in the dorsal root ganglia (DRG) of diabetic and non-diabetic DBA/2J mice in order to evaluate the biological relevance of literature-derived targets in the pathogenesis of diabetic neuropathy.http://jdrf.neurology.med.umich.edu/ROSDiabetes/). Fifty-three targets were significantly over-represented in the ROS-diabetes literature compared to randomly selected literature. These over-represented targets included well-known members of the oxidative stress response including catalase, the NADPH oxidase family, and the superoxide dismutase family of proteins. Eight of the nine selected genes exhibited significant differential expression between diabetic and non-diabetic mice. For six genes, the direction of expression change in diabetes paralleled enhanced oxidative stress in the DRG.SciMiner identified 1,026 ROS- and diabetes-related targets from the 1,154 biomedical papers (Literature mining compiled ROS-diabetes related targets from the biomedical literature and led us to evaluate the biological relevance of selected targets in the pathogenesis of diabetic neuropathy. Diabetes is a metabolic disease in which the body does not produce or properly respond to insulin, a hormone required to convert carbohydrates into energy for daily life. According to the American Diabetes Association, 23.6 million children and adults, approximately 7.8% of the population in the United States, have diabetes . The cosIn vivo and in vitro experiments implicate a number of enzymatic and non-enzymatic metabolic pathways in the initiation and progression of diabetic complications  AND "Diabetes Mellitus"[MeSH]). This query yielded 1,154 articles as of April 27, 2009. SciMiner, a web-based literature mining tool , was useThe NCBI Gene database provides links between Gene and PubMed. The links are the result of (1) manual curation within the NCBI via literature analysis as part of generating a Gene record, (2) integration of information from other public databases, and (3) GeneRIF (Gene Reference Into Function) in which human experts provide a brief summary of gene functions and make the connections between citation (PubMed) and Gene databases. For the 1,154 ROS-diabetes articles, gene-paper associations were retrieved from the NCBI Gene database. Non-human genes were mapped to homologous human genes through the NCBI HomoloGene database. The retrieved genes were compared against the SciMiner derived targets. Any genes missed by SciMiner were added to the ROS-diabetes target set.http://mimi.ncibi.org/) database  nor "Diabetes Mellitus"[MeSH]. For example, one of the ROS-diabetes articles (PMID: 18227068), was published in the Journal of Biological Chemistry, Volume 283, Issue 16. This issue contained 85 papers, 78 of which were not indexed with either "Reactive Oxygen Species"[MeSH] or "Diabetes Mellitus"[MeSH] indexed. One of these 78 papers was randomly selected as a background document. Three sets of 1,154 documents were selected using this approach and processed using SciMiner. Identified targets were confirmed by manual review for accuracy.ROS-diabetes targets were tested for over-representation against targets identified from the three background sets. Fisher's exact test was used to determine if the frequency of each target in the ROS-diabetes target set was significantly different from that of the background sets. Any targets with a BH adjusted p-value < 0.05 in at least two of the three comparisons were deemed to be an over-represented ROS-diabetes target. Functional enrichment analyses were performed on these over-represented ROS-diabetes targets as described above.th position and was additionally selected for inclusion in the test set.A subset of targets were selected for RT-PCR from the top 10 over-represented ROS-diabetes targets excluding insulin and NADPH oxidase 5 (NOX5), which does not have a mouse ortholog. Nitric oxide synthase 1 (NOS1), the main generator of nitric oxide, ranked at the 15DBA/2J mice were purchased from the Jackson Laboratory . Mice were housed in a pathogen-free environment and cared for following the University of Michigan Committee on the Care and Use of Animals guidelines. Mice were fed AIN76A chow . Male mice were used for this study.Two treatment groups were defined: control (n = 4) and diabetic (n = 4). Diabetes was induced at 13 weeks of age by low-dose streptozotocin (STZ) injections, 50 mg/kg/day for five consecutive days. All diabetic mice received LinBit sustained release insulin implants  at 8 weeks post-STZ treatment. Insulin implants were replaced every 4 weeks, at 12 and 16 weeks post-STZ treatment. At 20 weeks post-STZ treatment, mice were euthanized by sodium pentobarbital overdose and DRG were harvested as previously described .T)].The gene expression of the selected nine literature-derived ROS-diabetes targets in DRG was measured using real-time RT-PCR in duplicate. The amount of mRNA isolated from each DRG was normalized to an endogenous reference [Tbp: TATA box binding protein; Δ cycle threshold  and confirmed by manual review. Table The NCBI Gene2PubMed database, containing expert-curated associations between the NCBI Gene and PubMed databases, revealed 90 unique genes associated with the 1,154 ROS-diabetes papers  randomly selected from the complete HUGO gene set . The PPI network of the ROS-diabetes targets was significantly different from the randomly generated networks indicating their strong association with the topic "ROS and Diabetes". Table 6 randomlFunctional enrichment analyses of the 1,026 ROS-diabetes targets were performed to identify over-represented biological functions of the ROS-diabetes targets. After Benjamini-Hochberg correction, a total of 189 molecular functions, 450 biological processes, 73 cellular components and 341 pathways were significantly enriched in the ROS-diabetes targets when compared against all the HUGO genes  were generated using the same journal, volume and issue approach. The overlap among the three background sets in terms of documents and identified targets are illustrated in Figure Comparisons of the ROS-diabetes targets against these background sets revealed 53 highly over-represented ROS-diabetes targets as listed in Table Functional enrichment analyses of the 53 ROS-diabetes targets were performed to identify over-represented biological functions. Following Benjamini-Hochberg correction, a total of 65 molecular functions, 209 biological processes, 26 cellular components and 108 pathways were significantly over-represented when compared against all the HUGO genes . The mean ± SEM were 6.2 ± 0.3 for the non-diabetic control group and for 14.0 ± 0.8 for the diabetic group , indicative of prolonged hyperglycemia in the diabetic group . DRG werReactive oxygen species (ROS) are products of normal energy metabolism and play important roles in many other biological processes such as the immune response and signaling cascades -6. As meFifty-three targets were significantly over-represented in the ROS-diabetes papers when compared against three background sets. Depending on how the background set is defined, the over-represented targets may vary widely. An ideal background set would be the entire PubMed set; however, this is not possible due to limited access to full texts and intense data processing. An alternative method would be to use only abstracts in PubMed, but this may not fully represent the literature. Using only the abstracts, our target identification method resulted in 21 (39%) of the 53 key ROS-diabetes targets   and the F-measure /) before manual revision [One limitation of the current approach using literature mining technology is incorrect or missed identification of the mentioned targets within the literature. Based on a performance evaluation using a standard text set BioCreAtIvE  version 2 , SciMinerevision . In ordeThe present approach enabled us to collect a comprehensive list of ROS and diabetes related targets and led us to confirm the biological relevance to diabetic neuropathy of the selected ROS-diabetes targets. Using SciMiner to identify significantly enriched targets is applicable to other disease topics of interest by providing a more focused subset of literature for review and by highlighting targets common to multiple manuscripts.NAD(P): nicotinamide adenine dinucleotide (phosphate); AGE: advanced glycoation end-products; PKC: protein kinase C; ROS: reactive oxygen species; MAPK: mitogen-activated protein kinase; JNK: c-Jun amino-terminal kinase; SOD1: superoxide dismutase 1; SOD2: superoxide dismutase 2; CAT: catalase; XDH: xanthine dehydrogenase; NCF1: neutrophil cytosolic factor 1; NOS3: nitric oxide synthase 3; CYBA: cytochrome b-245 alpha; NOS1: nitric oxide synthase 1; ALS: amyotrophic lateral sclerosis; BioCreAtIvE: Critical Assessment of Information Extraction systems in Biology; MiMI: Michigan Molecular Interactions; KEGG: Kyoto Encyclopedia of Genes and Genomes; STZ: streptozotocin; GO: gene ontology; DRG: dorsal root ganglia; CNS: central nervous system; HGNC: HUGO (human genome organization) nomenclature committee; PPI: protein-protein interaction; SEM: standard error mean.The authors declare that they have no competing interests.JH participated in the study design, performed the literature mining and functional enrichment analyses, and drafted the manuscript. KAS participated in the study design and drafted the manuscript. ADS participated in the statistical analysis. YH carried out the quantitative RT-PCR assay. MP participated in the manuscript revision. DJS participated in the study design and manuscript revision. HVJ participated in the study design and manuscript revision. ELF participated in the study design and manuscript revision. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1755-8794/3/49/prepubThe list of 90 genes from the NCBI Gene2PubMed database for the ROS-Diabetes literature .Click here for fileThe list of 1,026 ROS-Diabetes targets.Click here for fileThe enriched Molecular Functions Gene Ontology Terms in the 1,026 ROS-Diabetes targets.Click here for fileThe enriched Biological Processes Gene Ontology Terms in the 1,026 ROS-Diabetes targets.Click here for fileThe enriched Cellular Components Gene Ontology Terms in the 1,026 ROS-Diabetes targets.Click here for fileThe enriched pathways in the 1,026 ROS-Diabetes targets.Click here for fileComparisons of target frequencies among three background sets.Click here for fileThe enriched Molecular Functions Gene Ontology Terms in the Over-represented 53 ROS-Diabetes targets.Click here for fileThe enriched Biological Processes Gene Ontology Terms in the Over-represented 53 ROS-Diabetes targets.Click here for fileThe enriched Cellular Components Gene Ontology Terms in the Over-represented 53 ROS-Diabetes targets.Click here for fileThe enriched pathways in the Over-represented 53 ROS-Diabetes targets.Click here for fileThe Key 53 ROS-Diabetes Targets Identifiable Using Only the Abstracts.Click here for file


SNCA gene expression and intracellular accumulation of the encoded α-synuclein (aSyn) protein are associated with the development of Parkinson disease (PD). To date, few enzymes have been examined for their ability to degrade aSyn. Here, we explore the effects of CTSD gene expression, which encodes the lysosomal protease cathepsin D (CathD), on aSyn processing.Elevated CTSD cDNA in dopaminergic MES23.5 cell cultures induced the marked proteolysis of exogenously expressed aSyn proteins in a dose-dependent manner. Unexpectedly, brain extractions, Western blotting and ELISA quantification revealed evidence for reduced levels of soluble endogenous aSyn in ctsd knock-out mice. However, these CathD-deficient mice also contained elevated levels of insoluble, oligomeric aSyn species, as detected by formic acid extraction. In accordance, immunohistochemical studies of ctsd-mutant brain from mice, sheep and humans revealed selective synucleinopathy-like changes that varied slightly among the three species. These changes included intracellular aSyn accumulation and formation of ubiquitin-positive inclusions. Furthermore, using an established Drosophila model of human synucleinopathy, we observed markedly enhanced retinal toxicity in ctsd-null flies.Over-expression of human ctsd gene mutations result in a lysosomal storage disorder that includes microscopic and biochemical evidence of aSyn misprocessing; and three, CathD deficiency facilitates aSyn toxicity. We therefore postulate that CathD promotes 'synucleinase' activity, and that enhancing its function may lower aSyn concentrations in vivo.We conclude from these complementary investigations that: one, CathD can effectively degrade excess aSyn in dopaminergic cells; two,  SNCA, as well as over-expression of the wild-type variant due to locus multiplication, cause autosomal-dominant forms of Parkinson disease (PD) . (A) A typical standard curve of serial dilutions of recombinant αS (mean of triplicate wells). (B) Following generation of MES-aSyn cells with three different concentrations of human SNCA cDNA , lysates were analyzed in 4 different dilutions, as indicated. Results show the mean absorbance signals of triplicates from a representative experiment. (C) Linear relationship between amount of SNCA-encoding cDNA transfected into MES23.5 cells and αS protein expression, as monitored by ELISA. Each lysate was analyzed in 4 different dilutions in triplicate. Linear regression of cDNA-to-protein was performed; results represent the mean aSyn values (μg) expressed from a representative experiment.Click here for fileSupplementary Figure 2 – Characterization of affinity-purified antibodies to α-synuclein using genotyped mouse brain. Whole brain extracts of genotyped mice were generated by lysis buffer that contained NP-40 and protease inhibitors [snca knock-out mice without a transgene  and mice that carry a human, wild-type SNCA transgene . Note the specific detection of full-length aSyn (16 kDa), and of 12.5 kDa and ~10 kDa truncated species of aSyn in SNCA-expressing mice.hibitors ; increasClick here for file


Tamoxifen may mediate its effect in early breast cancer in part via an oestrogen receptor (ER)-independent pathway by directly stimulating fibroblasts to produce the negative paracrine growth factor transforming growth factor (TGF)-beta. We have previously shown that secretion of this factor is induced 3-to 30-fold in human fetal fibroblasts in vitro, and by stromal fibroblasts in vivo following tamoxifen treatment of ER-positive and ER-negative breast cancer patients. Primary cultures of breast tumour fibroblasts have been exposed to tamoxifen for 48 h, and rates of secretion of TGF-beta 1 and TGF-beta 2 measured using a quantitative immunoassay. Fibroblast strains derived from malignant and benign tumours produced and secreted similar amounts of TGF-beta 1, but benign breast tumour fibroblasts secreted significantly higher levels of TGF-beta 2 compared with fibroblasts of malignant origin. Tamoxifen did not induce any consistent increase in TGF-beta secretion into the conditioned medium, but immunofluorescence analysis for the intracellular form of TGF-beta 1 revealed evidence of increased immunoreactive protein in tamoxifen-treated fibroblasts, which is localised to the nucleus. Therefore synthesis of TGF-beta 1 appears to be stimulated by tamoxifen, but increased secretion may be abrogated in vitro. Furthermore, using immunocytochemistry and transient transfection with an ER-responsive reporter construct, no ER was demonstrable in these fibroblasts supporting the proposed ER-independent paracrine pathway.


Nucleotide binding site-leucine rich repeat (NBS-LRR)-encoding genes comprise the largest class of plant disease resistance genes. The 149 NBS-LRR-encoding genes and the 58 related genes that do not encode LRRs represent approximately 0.8% of all ORFs so far annotated in Arabidopsis ecotype Col-0. Despite their prevalence in the genome and functional importance, there was little information regarding expression of these genes.erecta (Ler) after flagellin treatment. Several of these ten genes also showed altered expression after SA treatment, consistent with the regulation of R gene expression during defense responses and overlap between the basal defense response and salicylic acid signaling pathways. Enhancer trap analysis indicated that neither jasmonic acid nor benzothiadiazole (BTH), a salicylic acid analog, induced detectable expression of the five NBS-LRR-encoding genes and one TIR-NBS-encoding gene tested; however, BTH did induce detectable expression of the other TIR-NBS-encoding gene analyzed. Evidence for alternative mRNA polyadenylation sites was observed for many of the tested genes. Evidence for alternative splicing was found for at least 12 genes, 11 of which encode TIR-NBS-LRR proteins. There was no obvious correlation between expression pattern, phylogenetic relationship or genomic location of the NBS-LRR-encoding and related genes studied.We analyzed the expression patterns of ~170 NBS-LRR-encoding and related genes in Arabidopsis Col-0 using multiple analytical approaches: expressed sequenced tag (EST) representation, massively parallel signature sequencing (MPSS), microarray analysis, rapid amplification of cDNA ends (RACE) PCR, and gene trap lines. Most of these genes were expressed at low levels with a variety of tissue specificities. Expression was detected by at least one approach for all but 10 of these genes. The expression of some but not the majority of NBS-LRR-encoding and related genes was affected by salicylic acid (SA) treatment; the response to SA varied among different accessions. An analysis of previously published microarray data indicated that ten NBS-LRR-encoding and related genes exhibited increased expression in wild-type Landsberg Transcripts of many NBS-LRR-encoding and related genes were defined. Most were present at low levels and exhibited tissue-specific expression patterns. Expression data are consistent with most Arabidopsis NBS-LRR-encoding and related genes functioning in plant defense responses but do not preclude other biological roles. R) genes that are effective against diverse pathogens and pests, including bacteria, fungi, viruses, nematodes, and insects, have been cloned from both monocot and dicot plant species. These R genes can be divided into at least five classes based on the structure of their encoded proteins )EF-1α (769 to 4061 adjPPM) and ACT-2 (6–2925 adjPPM) (reference genes used in er (raw dClick here for file


Ciliated epithelium wasmagnified under an inverted microscope, and ciliary movement wasphoto-electrically measured. Ciliary activity was significantlyinhibited by 10−8 M lyso-PAF and could be restored. The effect oflyso-PAF was completely blocked by CV-6209, a specific PAFantagonist. The PAF concentration in the incubation medium oflyso-PAF was determined by radioimmunoassay, because PAF is a wellknown inhibitor of ciliary activity. PAF gradually increased andafter 20 min reached its maximal level. These findings indicated theexistence of an enzyme in the paranasal sinus mucosa, by whichlyso-PAF is converted to PAF, and that lyso-PAF can inhibit ciliaryactivity by means of PAF.The effect of lyso-PAF on ciliated cells was investigated


Male Wistar rats fed a normal laboratory pelleted diet, when treated s.c. with 1,2-dimethylhydrazine (DMH) 10 mg/kg/wk survived the 24-week experiment, showed no signs of chemical toxicity or macroscopic liver damage, and developed mainly large-bowel tumours. Conversely, male Wistar rats treated with 20 mg/kg/wk DMH did not survive the full term of the experiment and developed ascites, pleural effusions and nodular livers. They also developed more small-bowel tumours than large-bowel tumours. The relationship between the predominant site of tumour development and dosage of DMH was highly significant.Male Wistar rats fed with an all-liquid diet (Vivonex) and treated with 20 mg/kg/wk DMH behaved quite differently both in terms of survival and site of tumour development. These rats survived the full term of the experiment, showed no signs of chemical toxicity, experienced minimal liver damage and developed predominantly large-bowel tumours. The protection afforded by the all-liquid diet against DMH toxicity and small-bowel tumour induction was statistically highly significant.A series of blood tests with special reference to liver function confirmed the highly significant degree of protection against liver damage afforded by the all-liquid diet.Sections of liver from treated rats were examined, and a simple pathological scoring system was devised which showed a highly significant difference in liver histology between standard diet and liquid-diet rats treated with 20 mg/kg/wk DMH.The results strongly suggest an association between severity of liver damage from DMH and the subsequent development of small-bowel tumours. The all-liquid diet protected rats from liver damage and these rats developed significantly fewer small-bowel tumours.


This report describes our experience with two cases that were ultimately diagnosed as retroperitoneal liposarcoma using endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA). Case 1 is that of a 54-year-old woman with chief complaints of nausea and abdominal distention. Computed tomography (CT) and magnetic resonance imaging (MRI) revealed a large (15 cm diameter) tumor, which was significantly compressing the stomach and apparently occupied the entire left abdominal cavity. Although advanced primary gastrointestinal stromal tumor (GIST) or retroperitoneal tumor was inferred as the differential diagnosis, a definitive diagnosis was difficult using imaging alone. After EUS-FNA was done, the tumor was diagnosed histopathologically as high-grade liposarcoma. Case 2 is that of a 73-year-old man. Abdominal ultrasonography and CT showed a 6 cm diameter tumor within the pelvic cavity. The tumor had high MRI signal-intensity on both T1 and T2 images. Endorectal EUS showed a hyperechoic mass. The images suggested lipoma or liposarcoma containing lipoma-like components. Myxoid liposarcoma was revealed by subsequent EUS-FNA. Performing EUS-FNA was clinically useful for determining the subsequent therapeutic strategy in these cases where a tumor of unknown origin existed in the retroperitoneum. Endoscopic ultrasonography (EUS) is an important modality in the evaluati of luminal gastrointestinal diseases as well as diseases of the pancreas, gallbladder, and biliary ductal system. Endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) has been developed as a diagnostic tool using EUS . EUS-FNA Although liposarcoma is a malignant soft-tissue tumor that frequently occurs in extremities and retroperitoneum , it repoA 54-year-old woman who suffered from nausea and abdominal distention had consulted with her family doctor. She had no previous history of a related disease or these symptoms. Esophago-gastro-duodenoscopy (EGD) showed a subepithelial mass on the upper gastric corpus; an abdominal CT showed a huge tumor as a low-density mass occupying most of the left side of the abdominal cavity . She wasA 73-year-old man who showed a retroperitoneal mass on abdominal ultrasonography for health screening visited our hospital for further examination. Results of CT showed a 6 cm diameter low-density tumor within the pelvis . AbdominLiposarcoma is a kind of sarcoma that preferentially develops in the extremities (40%), retroperitoneal space (19%), and groin (12%), accounting for 9.8–16% of all sarcomas –7; with Histopathologic classification of liposarcomas was proposed by WHO classification  in 1969,As for treatment of liposarcomas, surgical therapy is the first option in resectable cases. Retroperitoneal liposarcoma has few clinical symptoms. Therefore, it often infiltrates into the surrounding organs. Consequently, the local recurrence rate is as high as 60% at five years. Decreasing the risk of recurrence requires en bloc excision including as much surrounding tissue as possible to prevent remnant tumor tissue , 13. ForEUS guided FNA has been applied for cytopathological diagnosis of a variety of lesions within and outside the gastrointestinal tract. Although there is a case of pancreatic metastasis of liposarcoma diagnosed by EUS-FNA , to our


Cell-surface glycoprotein of 3 rat hepatoma strains and late-embryonic liver was metabolically labelled in vivo with [3H]- or [14C]-fucose. Trypsinization of the cells and exhaustive pronase digestion of combined hepatoma-liver trypsinates followed by gel filtration over Sephadex-Biogel mixtures, yielded elution profiles that contained more early-eluting (high-mol.-wt.) glycopeptides for hepatomas than for liver. At least 3 factors were identified which acted to augment the fraction of early-eluting tumour glycopeptides: (a) increase of neuraminidase-sensitive sialic acid, (b) increase of neuraminidase-insensitive sialic acid that was sensitive to mild HCl hydrolysis, and (c) presence of sugar sulphate groups contributing to a restricted extent, relative to possible unknown factor(s). Whether (a), (b) or (c) operated depended on the hepatoma strain or its mode of growth. Notwithstanding these differences in the nature of the increase in early-eluting glycopeptides, the increase itself appears not to be due to growth per se, nor to an embryonic expression, but rather may serve as a marker of tumourigenicity.


During 1998, a marked increase (35 cases) in human gastroenteritis due to Salmonella Blockley, a serotype rarely isolated from humans in the Western Hemisphere, was noted in Greece. The two dominant multidrug-resistance phenotypes (23 of the 29 isolates studied) were associated with two distinct DNA fingerprints, obtained by pulsed-field gel electrophoresis of genomic DNA.


While the compound shows no activity against K-562 cells, there isevidence for some cytotoxicity against SK-OV-3. The compound is much less effective than cisplatin, and itslimited solubility restricts the useable concentration range.A simple method for the preparation of


Inaddition, several long-term catfish monocyte lines have been found to function as efficientAPC with autologous but not allogeneic responders. Subsequent separation of theresponding PBL into sIg- (T-cell-enriched) and B (sIg+) cell subsets showed that both underwentproliferative responses to antigen-pulsed and fixed APC. Moreover, allogeneic cellsused as APC were found to induce only strong mixed leukocyte reactions without specific invitro antibody production. Initial attempts at identifying the immunogenic region(s) of theprotein antigens for catfish indicated there are two such regions for pCytC, namely, peptides66-80 and 81-104.This work was undertaken to investigate whether or not antigen processing and presentationare important in channel catfish in vitro secondary immune responses elicited withstructurally defined proteins, namely, pigeon heart cytochrome C (pCytC), hen egglysozyme, and horse myoglobin. The use of


Danshen, the dried root of Salvia miltiorrhiza, is a Chinese medicine used to promote blood flow and treat vascular disease. The present article reviews the pharmacological effects of Danshen on cerebral infarction and possible interactions between Danshen and Western drugs. Danshen may reduce or prolong the development of atherosclerosis and may have anti-hypertensive and anti-platelet aggregation effects, which prevent cerebral infarction. Danshen may enhance endogenous anti-oxidative enzyme activities such as the expression of endothelial nitric oxide synthase and may scavenge oxygen free radicals. Prevention and treatment of cerebral infarction by Danshen involves multiple pathways, including anti-atherosclerosis, anti-hypertension, anti-platelet aggregation, anti-inflammatory and anti-oxidative effects. Danshen, the dried root of Salvia miltiorrhiza, is used in Chinese medicine to treat vascular disease. According to Chinese medicine theory, Danshen promotes blood flow and resolves blood stasis. Among stroke patients, 80% suffer from cerebral infarction and 20% cerebral hemorrhage [morrhage . Cerebramorrhage . Anti-thmorrhage .Danshen or its ingredients reduce the infarction volume, the action mechanism of Danshen remains obscure [Danshen contains the lipid-soluble tanshinone I (Tan I), tanshinone IIA (Tan IIA), cryptotanshinone and dihydrotan-shinone as well as the water-soluble danshensu and salvianolic acid B  [Danshen inhibits platelet aggregation and promotes fibrinolysis [While  obscure -8. Dansh  . Dansheninolysis ,11.Danshen in the prevention and treatment for cerebral infarction. The key words used for search in the PubMed library were 'Danshen' or 'Salvia miltiorrhiza' combined with 'cerebral infarction', 'ischemic' or 'cerebral ischemia'.Covering literature between 1990 and 2009, this article reviews basic and clinical studies on Danshen  inhibit the production of interleukin-12 (IL-12) induced by lipopolysaccharide (LPS)-activated macrophages and the production of interferon-γ induced by keyhole limpet hemocyanin-primed lymph node cells [Danshen or Sal B component of Danshen inhibits TNF-α-induced expression of VCAM-1 and TNF-α-induced activation of NF-κB in human aortic endothelial cells, suggesting that both SME and Sal B posses anti-inflammatory properties and are closely associated with atherogenesis because leukocytes migrate into the vessel wall in the early stage of atherogenesis [Danshen suppresses the adhesion rate of neutrophils stimulated by TNF-α, and also inhibits adhesion of neutrophils stimulated by N-formyl-methionyl-leucyl-phenylalanine (fMLP) [Danshen inhibits TNF-α-induced up-regulation of E-selectin, intracellular molecule-1 (ICAM-1) and VCAM-1 expression [Danshen treatment lowers plasma viscosity, erythrocyte aggregation and fibrinogen levels in rats with traumatic brain injury [Danshen reduces the aortic atherosclerotic lesion area and inhibits inflammatory serum marker of CRP and monocyte chemotactic protein (MCP-1) [Danshen reduces the serum levels of total cholesterol and triglyceride in low density lipoprotein receptor (LDLR) +mice via the inhibition of the inflammatory markers CRP and MCP-1 [Danshen play roles in the prevention of cerebral infarction.Atherosclerosis is caused by endothelial damage, which may lead to platelet aggregation and the release of platelet factor, resulting in the proliferation of smooth muscle in the arterial intima. The process of atherosclerosis comprises inflammation and involves both innate and adaptive immunity . The tande cells . Tanshinde cells . As IL-1de cells . Tumor nogenesis . Aqueouse (fMLP) . Aqueouspression . Danshenn injury . Tan IIAn injury . C-reactn injury ,19. Tan n injury . Tan IIAn injury . Triterp (MCP-1) . Moreovend MCP-1 . Triterpnd MCP-1 . In summDanshen alleviates hypertension in the two-kidney/one-clip (2K1C) Goldblatt renovascular hypertensive rats model and reduced the activity of serum angiotensin converting enzyme and the levels of serum aldosterone [Danshen. MTB70 (0.7-175 mg/kg) reduces blood pressure more than the aqueous extract of Danshen with same volume in a saline- or phenylephrine (2.5 μg/kg bolus injection and followed by 100 μg/kg at 0.13 ml/hr injection continuously) -induced hypertension model in rats; However this rapid and potent anti-hypertensive effect is short-lived [Danshen include inhibition of angiotensin converting enzyme and generation of eNOS and/or vasodilatation.Hypertension is a risk factor for cerebral infarction. Maintaining blood pressure within the ideal range has recently been recognized as an important principle in the prevention and treatment of ischemic cerebral infarction. Aqueous extract of osterone ,24. MTB7rt-lived . Tanshinrt-lived . OverallDanshen inhibit platelet aggregation induced by adenosine diphosphate (ADP) in vitro in rabbit plasma [in vivo and in vitro, probably through changing protein expression [Danshen decreases the malondialdehyde (MDA) levels of platelets and increases the superoxide dismutase (SOD) activity of platelets to inhibit platelet aggregation in pulmonary thromboembolism induced by collagen and adrenaline in mice [Danshen extract, inhibits platelet aggregation induced by arachidonic acid or ADP in humans and rabbits [Danshen decreases the concentration of intra-platelet free calcium, which is closely associated with platelet aggregation and release [Danshen exhibits anti-platelet aggregation effects via multiple pathways, e.g. the inhibition of intra-platelet calcium and anti-oxidant activities.Anti-platelet aggregation, which involves platelet adhesion, is widely used to treat acute ischemic cerebral infarction . Platelet plasma . Salviant plasma . Salvianpression . Danshen in mice . The 764 rabbits . Danshen release . OverallDanshen inhibits superoxide generation by microglia in primary microglia cell cultures from rat brains [Danshen reduces CD18 and CD11b immunoreactive cells in the peri-vascular region and inhibits leukocyte infiltration and neuronal death in the cerebral infarction region in the ischemia-reperfusion MCAo rat model. As leukocytes are mediated via the combination of surface receptors of CD18 and CD11b and intracellular adhesion molecules of endothelium, the neuroprotection capability of Danshen may be associated with the inhibition of leukocyte adherence to the endothelium [Danshen on inflammatory response in cerebral ischemia and reperfusion injury in the MCAo rat model indicates that pre-treatment with Tanshinone IIA reduces the cerebral infarction area, neurological deficit score and cerebral edema. As pre-treatment with Tanshinone IIA reduces TNF-α, the myeloperoxidase (MPO) marker of leukocytes, E-selectin and ICAM-1 in ischemic brain tissue and serum IL-8, Danshen may inhibit inflammatory responses to reduce brain damage induced by ischemia-reperfusion injury in rats [Danshen dripping pill (DDP) is composed of Danshen, Panax notoginseng and Dryobalanops camphor. A controlled pilot study (106 patients) found that the recurrent rate in the DDP group was 9.6% compared with 24.1% in the non-DDP group. The serum CRP levels decreased from 2.33 to 1.50 mg/L in the DPP group were greater than those decreased from 2.21 to 1.77 mg/L in the non-DDP group [Danshen has anti-inflammatory activity and may improve cerebral infarction and provide neuroprotection.Inflammatory responses are critically important to a patient after brain ischemia/reperfusion injury. The pro-inflammatory cytokines IL-1β, TNF-α and IL-6 increase after an ischemia attack, which enhances the expression of adhesion molecules including ICAM-1 and P-selectin, leading to brain edema and neuronal death ,36. In at brains . Danshenothelium . A study in rats . The DanDP group . OverallDanshensu and salvianolic acid B are phenolic acids of Danshen, both of which have scavenging activities towards free radicals of hydroxyl, 1,1-diphenyl-2-picryl-hydrazyl (DPPH), 2-azino-bis(3-ehtylbenzthiazoline-6-sulfonic acid) (ABTS) free radicals, hydrogen peroxidase and superoxide anion in human umbilical vein endothelial cells [Danshen reduces the cerebral infarction volume and the neurological deficit score and reduces luminal-chemiluminescence counts [Danshen may be associated with its free radical scavenging activities in ischemia-reperfusion injured rats [Danshen, reduces the focal infarct volume, neurological deficit and apoptosis in the MCAo rat model. As apoptosis is a major pathway leading to neuronal death, Tan IIB is neuroprotective [Danshen reduced an increase of cerebral NO and MDA 30 minutes prior to occlusion and that (2) pre-treatment with Danshen increased a significantly decreased SOD activity. NO mediates glutamate neurotoxicity and the inhibition of NO synthase may prevent the development of brain edema. SOD is a scavenger of superoxide anion and the levels of MDA may reflect the degree of lipid peroxidation. Thus Danshen is neuroprotective in this occlusion and reperfusion rat model [Danshen in rat brain, reduces cerebral infarction volume, brain edema and neurological deficit in the MCAo rat model. Moreover, ND309 reduces brain tissue MDA and increases the reduction of ATP induced by ischemia-reperfusion injury [Danshen reduces the neurological deficit score and the levels of MDA and increases SOD activities in the MCAo rat model [Danshen may reduce brain edema and MDA concentration of cerebral cortex and hippocampus region in an ischemia-reperfusion injured rat model while Danshen may increase levels of catalase, SOD, GSH and ATP of the cerebral cortex and hippocampus region [Danshen demonstrates neuroprotective effects that are closely associated with anti-oxidative action.Nitrogen and oxygen free radicals involved in the production of reactive oxygen species (ROS) play an important role in brain damage during the reperfusion period following ischemia ,44. Thesal cells . Our pree counts . As lumired rats . Intra-aotective . Damage otective ,49 foundat model ,49. Isopn injury  of mitocn injury . Danshenat model . Pretreas region . In summDanshen inhibits platelet adhesion and aggregation and suppresses the formation of thromboxane A 2 [Danshen and warfarin can cause bleeding and prolong prothrombin time or the International Normalized Ratio (INR) [Danshen and warfarin should be avoided or closely monitored [Danshen displaces the binding between aspirin and protein, thereby increasing free aspirin concentration in serum [Danshen has digoxin-like immunoreactivity leading to a false interference of digoxin concentration [Warfarin is an anticoagulant used to prevent atrial fibrillation, valvular heart disease, ischemic stroke and deep venous thrombosis . Danshenxane A 2 . The intio (INR) . Co-admionitored . Tanshinonitored . Aspirinin serum . Atrial ntration .2O 2 [Panax notoginseng (Sanqi) is also used to treat vascular disorders. Danshen is stronger than Panax notoginseng in scavenging activity of superoxide anion, hydroxyl and DPPH radicals, yet it is weaker than Panax notoginseng in the scavenging activity of hydrogen peroxide and ferrous ion chelating activity [Sanqi demonstrates strong ferrous ion chelating activity and strong scavenging activities of hydrogen peroxide and hydroxyl radicals, it is weak in the scavenging activities of superoxide anion and DPPH radicals. Therefore, the scavenging capabilities of Danshen and Sanqi are quite different among the various free radicals [The extracts of Ginkgo biloba L. leaves (Egb 761), with free radical scavenging activity, reduce the size of cerebral infarction and improve neurological behavior in rats with permanent and transient MCAo ,58. Egb 2O 2 . Panax nactivity . While Sradicals .Danshen involves multiple pathways, including anti-atherosclerosis, anti-hypertension, anti-platelet aggregation, anti-inflammatory and anti-oxidative effects ; ND-309: isopropyl-β,--α-hydroxypropanoate; Egb 761: the extracts of Ginkgo biloba L. leavesThe authors declare that they have no competing interests.THL searched the literature, organized the data and wrote the manuscript. CLH analyzed the data and revised the manuscript. Both authors read and approved the final version of the manuscript.Danshen for prevention and treatment of cerebral infarctionPossible pharmacological actions of . Supplemental tableClick here for file


The chemistry of the mitomycin C-related drug indoloquinone EO9 would suggest that its mechanism of action is likely to involve DNA damage after reductive activation. The ability of this agent to induce DNA damage in intact cells has been examined using alkaline filter elution. After treatment with pharmacologically relevant concentrations of EO9, both DNA strand breaks and interstrand cross-links were detected in rat Walker tumour cells and human HT29 colon carcinoma cells. These cell lines express relatively high levels of DT-diaphorase (NAD(P)H: quinone acceptor oxidoreductase), which is believed to be involved in EO9 activation. The extent of DNA damage was increased by approximately 30-fold under hypoxia in BE colon carcinoma cells that express non-functional DT-diaphorase, but this dramatic hypoxia enhancement was not seen in HT-29 cells. These data are consistent with cytotoxicity studies that indicate that DT-diaphorase appears to be important in EO9 activation under aerobic conditions, but other enzymes may be more relevant under hypoxia. The involvement of DT-diaphorase in DNA damage induction was further investigated using cell-free assays. DNA cross-links were detectable in plasmid DNA co-incubated with EO9, cofactor and DT-diaphorase but not in the absence of this enzyme. In contrast, using a Taq polymerase stop assay, monofunctional alkylation was detected in plasmid DNA without metabolic activation, although the sequence selectivity was altered after reduction catalysed by DT-diaphorase.


Twenty cases of interstitial pneumonia secondary to treatment with granulocyte colony-stimulating factor (G-CSF) were reviewed. Their interstitial pneumonia had the following features: (a) it occurred predominantly in patients aged 60 years or older; (b) it was prevalent among patients with haematological malignancies, particularly non-Hodgkin's lymphoma; (c) in all patients G-CSF was given after anti-cancer agents with potential to affect the lungs; (d) at the onset, many patients had symptoms such as dyspnoea and fever; and (e) the leucocyte (neutrophil) count as well as lactate dehydrogenase (LDH) and C-reactive protein (CRP) levels were usually higher than normal at the onset. These findings indicate that, when G-CSF is used in combination with pneumotoxic anti-cancer agents, respiratory function should be monitored before and during treatment. If the leucocyte (or neutrophil) count and/or LDH and CRP increase suddenly in association with dyspnoea and fever during administration of G-CSF, interstitial pneumonia should be suspected. Accordingly, a chest radiograph and pulmonary functional tests should be performed promptly. If a diagnosis of interstitial pneumonia is made, steroid pulse therapy should be commenced immediately.


This study shows that human lymphocytes markedly decrease chloramines (long-lived oxidants) generated by polymorphonuclear neutrophils (PMN) after stimulation by phorbol-myristate-acetate or opsonized zymosan. In a cell-free model, reduced glutathione (GSH) scavenged chloramines, giving rise to oxidized glutathione (GSSG). In the cell system, treatment of lymphocytes with autologous PMN-derived chloramines induced a profound decrease in their total and reduced glutathione (GSH) content and markedly inhibited their proliferate responses to concanavalin-A and, to a lesser extent, phytohaemagglutinin. It is concluded that (i) lymphocytes may play a defensive role against phagocyte-derived oxidative stress by scavenging chloramines, and (ii) as this effect which is mediated by GSH affects lymphocyte proliferative responses, it may help to elucidate the still obscure mechanisms of oxidative stress associated immunodeficiency.


P and gut inflammation, changes in substance P levels were evaluated in a chronic model of ileitis in response to three anti-inflammatory agents with distinct mechanisms of action. The agents were the prostaglandin E1 analogue misoprostol , the nitric oxide synthase inhibitor NG-nitro-L-arginine methyl ester  and the leumedin, N-(fluorenyl-9-methoxycarbonyl)-L-leucine . Ileitis was induced by a transmural injection of trinitrobenzene sulphonic acid (TNBS 30 mg/kg in 50% ethanol) into the distal ileum of guinea-pigs. All anti-inflammatory therapies were introduced after TNBS administration and continued until day 7, when guinea-pigs were killed. Ileal substance P levels were measured by radioimmunoassay, and granulocyte infiltration was quantified by myeloperoxidase (MPO) activity. Protein and nitrite (an index of nitric oxide formation) levels in a luminal saline lavage were quantified in all groups. TNBS ileitis caused a marked reduction in ileal substance P content and increased MPO activity, protein and nitrite secretion. The nitric oxide synthase inhibitor, L-NAME, completely restored all parameters to baseline. Misoprostol attenuated the granulocyte infiltration and exacerbated protein leak but had no effect on substance P levels. In contrast, NPC 15199 had no effect on granulocyte infiltration but normalized substance P levels and protein leak. Only L-NAME and NPC 15199 blocked the TNBS induced increase in nitrite levels. These results suggest that the regulation of granulocyte infiltration in this model is unrelated to changes in substance P levels. Inhibition of nitric oxide synthase was the most effective therapeutic strategy in TNBS ileitis but the precise interactions between nitric oxide and the enteric nervous system during inflammatory states remain to be defined.To understand the interactions between substance


For years an assumption was made that C-peptide, abyproduct of insulin biosynthesis, possessed no appreciablephysiologic role. As other contributions in this volumeamply testify, the time has come to re-evaluate that notion.C-peptide either directly through interaction with its specificcell-surface receptor or indirectly through an interactionwith a related membrane entity, exerts a unique effecton several intracellular processes.We review here results ofstudies attempting to elucidate such molecular effects ofC-peptide in different cell systems and tissues. Lacking apurified C-peptide receptor, we also demonstrate C-peptideeffects on distinct elements of the insulin signal transductionpathways.


Although DCs can be directly infected by certain strains of HIV-1, productive infection of DCs is not required during trans-infection; instead, DCs capture and internalize infectious HIV-1 virions in vesicles for later transmission to CD4 T cells via vesicular exocytosis across the infectious synapse. This model of sequential endocytosis and exocytosis of intact HIV-1 virions has been dubbed the “Trojan horse” model of HIV-1 trans-infection. While this model gained rapid favor as a strong example of how a pathogen exploits the natural properties of its cellular host, our recent studies challenge this model by showing that the vast majority of virions transmitted in trans originate from the plasma membrane rather than from intracellular vesicles. This review traces the experimental lines of evidence that have contributed to what we view as the “rise and decline” of the Trojan horse model of HIV-1 trans-infection.To ensure their survival, microbial pathogens have evolved diverse strategies to subvert host immune defenses. The human retrovirus HIV-1 has been proposed to hijack the natural endocytic function of dendritic cells (DCs) to infect interacting CD4 T cells in a process termed  Dendritic cells (DCs) play a central role in initiating the adaptive immune response that counters pathogen infection. Immature DCs patrol the peripheral mucosal tissues, searching for unwanted intruders. Once a pathogen is sensed, captured, and internalized, DCs undergo a maturation process and migrate to the regional lymph nodes. Meanwhile, the internalized pathogens are processed into antigenic peptides, and co-stimulatory molecules are expressed on the cell surface, readying these professional antigen-presenting cells for effective T-cell stimulation trans-infection.Pathogens have evolved various means to escape the host immune response by subverting the function of DCs. HIV-1 excels in this capacity. Like many other microbial pathogens, HIV-1 interferes with Toll-like receptor signaling, impairing the secretion of antiviral and inflammatory cytokines needed for the development of an effective immune response trans-infection in vivo is lacking, numerous in vitro observations suggest that such capture and transfer of virions to permissive cells is advantageous for the virus, in particular when quantities of infectious particles are limiting. In early studies of ex vivo tissue explants, most HIV-1 replication was observed within DC–T-cell conjugates Although direct evidence of in trans to interacting CD4 T cells trans-infection involved internalization of virions into an endocytic compartment trans-infection in which exocytosis of virion-laden vesicles into the synapse promoted highly efficient infection of CD4 T cells.The mechanism underlying this DC-dependent enhancement of HIV infection remained unknown until the discovery of DC-specific intercellular adhesion molecule-grabbing non-integrin (DC-SIGN) trans-infection of T cells trans-infection of T cells by DCs derived in vitro from monocytes (MDDCs) involves DC-SIGN DC-SIGN, the most-studied C-type lectin receptor that captures HIV-1 virions, is a calcium-dependent lectin that binds the HIV envelope with an affinity similar to that of CD4 trans-infection mediated by mannose receptors in DCs is lacking, but in macrophages, these receptors appear to play an important role in the trans-infection of CD4 T cells in transtrans-infection mediated by DC-SIGN on MDDCs, suggesting that langerin and DC-SIGN may, in fact, mediate opposite fates for HIV-1 virions. However, caution is warranted when comparing results from ex vivo LCs and in vitro derived DCs. LCs derived in vitro from CD34-expressing precursor cells, like MDDCs, mediate efficient trans-infection of T cells despite expression of langerin trans-infection Other C-type lectin receptors, such as langerin and mannose receptors, are at least equally important for gp120 binding to epithelial DCs As part of their normal function, DCs internalize pathogens and process proteins from these organisms into small antigenic peptides for subsequent presentation to CD4 T cells on MHC-II receptors. Typically, immature DCs display high levels of endocytic capacity while mature DCs are characterized by efficient antigen processing and presentation. Early in vitro and ex vivo studies supported the notion that DCs internalize structurally intact HIV-1 virions into large vacuolar structures In mature MDDCs, the compartment harboring HIV-1 virions shares certain features with the late endosome or the multivesicular body (MVB), but other features differentiate these structures from classical late endosomes or lysosomes trans-infection of CD4 T cells Upon interaction with T cells, HIV-1-loaded MDDCs redistribute virion-containing vesicles to the DC–T-cell junctions trans-infection, we tested the effects of soluble CD4 (sCD4). This agent selectively neutralizes gp120 on surface-bound virions while not altering internalized virions. To our surprise, sCD4 completely inhibited HIV-1 trans-infection trans-infection. To exclude possible unappreciated effects of sCD4, we also inactivated surface-bound virions with pronase. Again, trans-infection was abrogated by the selective incapacitation of surface-bound HIV-1 virions in both MDDCs and CD34-derived LCs. These findings implicating surface virions in trans-infection .A major drawback of protease treatment of the cell surface is its relative lack of specificity for bound HIV-1 virions. Key components of the infectious synapse could be digested, resulting in altered or delayed synapse formation. The use of more specific membrane-impermeable inhibitors, such as sCD4 and HIV-1-neutralizing antibodies, avoids these complications. Two studies with these agents have appeared trans-infection in the context of opsonized HIV-1 virions.Recently, a twist on the Trojan horse model emerged in a study suggesting that HIV-1 virions in immune complexes may recover their infectivity after capture by immature MDDCs and release of the bound antibodies within the acidified endocytic compartment trans-infect T cells. Such factors include the state of DC activation and maturation, the time elapsing between virion capture by DCs and contact with interacting T cells, and the nature of receptors that mediate binding of HIV-1 virions.The fact that virions are almost exclusively transmitted from the DC surface implies that virion internalization is chiefly a dead end for infectious virions. Several factors influencing the internalization of HIV-1 virions might affect their likelihood to trans-infect T cells External molecules are taken up in DCs via multiple pathways, including phagocytosis, macropinocytosis, and receptor-mediated endocytosis via clathrin-coated pits and caveolae. DC differentiation and maturation are associated with a decline in endocytic activity, particularly macropinocytosis trans-infection. In initial studies, virions captured by Raji-DC-SIGN and MDDCs retained their infectivity for several days trans-infection for only a few hours after capture trans-infection trans-infection trans-infection, as seen in macrophages trans-infection of T cells by DCs. Interestingly, the use of inhibitors that impair intracellular trafficking and/or acidification of the endosomes appear to exert only minimal effects on trans-infection trans-infection trans-infection. Conversely, expression of Nef in MDDCs and neutralization of CD4 by antibodies decrease available surface CD4 and are associated with increases in trans-infection. Collectively, these studies suggest that virion internalization negatively regulates trans-infection and are consistent with surface-bound virions forming the principal source of virions mediating trans-infection.The time elapsing between virion capture by DCs and contact with interacting T cells may also affect the efficiency of trans-infection of T cells by DCs involves primarily surface-bound virions argues that future research should be refocused on how HIV-1 hijacks the plasma membrane rather than the intracellular trafficking pathway as suggested by the original Trojan horse model. Unraveling how these virions are recruited to the infectious synapse is crucial. The presence of C-type lectin receptors in lipid rafts in trans will be important. Finally, our new model suggests that in vivo transmission of virions captured by DCs to T cells is likely to be far more sensitive to attachment inhibitors and neutralizing antibodies than previously anticipated. Only time will tell whether this fact can be therapeutically exploited.The finding that


A large number of cells containing subunit a of blood coagulation Factor XIII (FXIII) was detected by immunoperoxidase staining in lymph nodes with Hodgkin's disease. These relatively large, multipolar, mononuclear cells were often found in the immediate vicinity of malignant Hodgkin's cells. Intensive characterization of these cells carried out by immunofluorescent and enzymecytochemical techniques in double- and triple-labelling systems on the same sections clearly demonstrated that they represent tumour-associated macrophages (TAMs). FXIII containing-cells showed alpha-naphtyl acetate esterase (ANAE) positivity, and were labelled by monoclonal anti-Leu M3 antibody, a monocyte/macrophage marker, but not at all or only very weakly by anti-HLA-DR. Neither alkaline phosphatase (ALP) nor adenosine triphosphatase (ATPase) activity could be detected in these cells and surprisingly, they were consistently negative for acid phosphatase (AcP) as well. The presence of FXIII subunit a in tumour-associated macrophages suggests that this cell type might have an important role in the stabilization of fibrin deposits around tumour cells.


Although recruiting persons with dementia into research is challenging enough, finding those who live-alone in the community is even more difficult. Consequently, live-alone persons with dementia are often overlooked and/or deliberately excluded from inquiry despite calls for more inclusive approaches to dementia research. Based on enrollment strategies from an interview-based protocol recruiting 120 live-alone persons with dementia, our National Institute on Aging- funded study identified five domains of gatekeepers imperative to gaining access to community-dwelling, live-alone persons with dementia: 1) housing , 2) data proprietors , 3) institutional , 4) kin (including fictive kin), 5) clinical . In addition, gatekeeper domains are multilayered and serve distinct roles in both facilitating and hindering access to and enrollment of this under-researched vulnerable population. Analysis of our recruitment efforts contribute significant insights into how the dementia research community may engage the various domains of community gatekeepers, providing direction for current and future social science research.


However, the use of circulating free miR-21 as a non-invasive biomarker for diagnosis and therapeutic monitoring in breast cancer is not well established. We quantified the levels of circulating miR-21 expression and analyzed their correlation with clinicopathological variables and progression-free survival. Aberrant patterns of microRNA expression have been highlighted as a potential clinical biomarker in breast cancer as the most frequent cancer among women that contributes nearly a quarter of total cancer incidence in 2018. Upregulation of microRNA-21  was used to analyzed miR-21 expression. This initial study included a cohort of 102 breast cancer patients of different subtypes and clinicat stages. We also included 15 unrelated healthy women. Venous blood from patients was collected at diagnosis and after treatment of surgery and chemotherapy. miR-21 was significantly elevated in breast cancer patients compared to healthy women . Significant reduction of miR-21 expression was observed in breast cancer patients after completion of surgery and chemotherapy . MiR-21 expression was higher in breast cancer patients younger than 40-year-old but was not significantly different according to different histopathological grades and clinical stages at diagnosis. Patients with upregulation of circulating miR-21 were associated with poor progression-free survival  test, p=0.049).Expression of circulating MiR-21 expression was upregulated in breast cancer patients and might serve as a therapeutic monitoring marker. Breast cancer has posed as a major public health challenge because the incidence is continuously increasing during the past decades . With more than 2 million new cases in 2018, breast cancer is the most frequent cancer diagnosed among women worldwide including in Indonesia . Although significant improvement in the diagnosis and treatment has recently been achieved, breast cancer-related mortality and morbidity are relatively high particularly in low- and middle-income countries . In Indonesia, breast cancer patients often present in late stages due to lack of public awareness, screening participation, as well as access for diagnosis and treatment . In addition, breast cancer is a heterogeneous disease showing various phenotypes, subgroups, and different responses to treatment . Cancer antigen 15-3 (CA-153) is currently the most common biomarker used in breast cancer although the relevance into the clinical practice, outcome, and monitoring of relapse are still controversial due to lacking of sensitivity and specificity . Therefore, investigation for finding novel non-invasive biomarker in breast cancer is required.MicroRNAs are small non-coding RNAs regulating gene expression post-transcriptionally through direct interaction with the messenger-RNA targets . Deregulation of particular microRNA panels has been associated with diagnostic and prognostic values of various cancers including breast cancer . Patterns of microRNA expression have also been extrapolated in breast cancer sub-classification  and metastasis detection . In the rapidly expanding field of cancer stem cells, several microRNAs have also been reported in maintaining the stemness phenotypes . We recently reported interaction of a polymorphism with microRNA binding and downregulation of target gene expression as well as their correlation with tumor size and lymph node infiltration in estrogen receptor (ER)-negative breast cancer . In addition, microRNAs are also released from tumor cells into blood circulation through lipid vesicles, microvesicles, exosomes, and apoptotic bodies . Circulating microRNAs typically appear in a complex formation with ribonucleoprotein or high-density lipoprotein owing their stability against endogenous RNase degradation . Detection of circulating microRNAs has also gained interest in its potential application as a non-invasive biomarker in cancer . MiR-21 is one of the most frequently reported oncogenic microRNA in various solid tumors in which the upregulation is commonly related with adverse outcome . Functional studies have also shown that miR-21 upregulation is able to induce cell proliferation, migration, and inhibit apoptosis . Validated miR-21 target genes are mostly tumor suppressor genes including PTEN  and PDCD4 . Although prognostic and diagnostic values of miR-21 in breast cancer have been highlighted in some reports , the debate regarding the cost-effective measurement and practical implementation remains exist. In addition, exploration of miR-21 alteration after therapy is underreported. In this study, we investigated the potential use of circulating miR-21 as a biomarker for diagnosis, prognosis and therapeutic monitoring.Patient cohortBlood samples were collected in the periode of 2014-2015 from 102 breast cancer patients at diagnosis, 40 breast cancer patients after completion of surgery and chemotherapy, and 15 healthy women without a history of benign and malignant tumors. Patients and participants who were older than 18 years old, were confirmed diagnosis by histopathology, and were able to provide inform concent, were then recruited. Vulnerable patients and participants such as with dementia and terminally-ill patients were excluded. Tumor stage and clinicopathology variables were collected at the time of diagnosis. Follow up of patients was performed according to the local clinical recommendation. Progression-free survival was defined as the period from the time at diagnosis to any relapse or local/regional/distant disease spreading observed until June 2017. Sample size was estimated using effect size of Cohen’s approach with α=0.05, moderate effect size of 0.5, and statistical power of 0.9. The healthy individuals used as controls were selected randomly to represent the population. Protocol of the study was approved by the Medical and Health Research Ethics Committee of the Faculty of Medicine, Universitas Gadjah Mada, Indonesia (601/EC/2014). All participants gave written informed concent prior to participating in this study.RNA isolation from plasmaoC until analysis. Total RNA was isolated from 200 µL plasma using miRCURYTM RNA Isolation Kit-Biofluid  according to the manufacturer’s recommendation. The plasma was mixed with Lysis and Protein Precipitation Solution. After centrifugation 12,000 rpm, the supernatant was mixed with 270 µL isopropanol and was then applied into microRNA Mini Spin Column (Exiqon). After washing steps, total RNA was eluted using 25 µL RNA-se free water. For internal control during RNA extraction, we used spike-in of Sp6 into each plasma sample. The peripheral venous blood samples were collected using EDTA-vacutainer collection tubes and centrifugated at 1500 rpm for 10 minutes. The supernatant plasma was stored at -80Quantification of miR-21 expressionMiR-21 expression levels were measured using ExiLent SYBR Green master mix  and specific primers for miR-21 , Sp6 , and MIR-16 . Enriched RNA from plasma was reverse transcribed using Universal cDNA Synthesis kit II, 8-64 rxns  in a total reaction volume of 20 µL. Statistical analysismiR-21 was normalized using spike-in Sp6 and miR-16 to further be compared with the expression in healthy individuals. Fold change expression was calculated using the 2-ΔΔCq method. The expression levels of miR-21 between breast cancer patients and healthy women were compared using the Mann-Whitney-U test. The miR-21 expression levels at diagnosis and after treatment were compared and the correlation between clinicopathologic variables with miR-21 expression was also be compared with Mann-Whitney-U tests. Upregulation was determined as mean of miR-21 expression + 2xSD. Kaplan-Meier plot and Log-rank (Mantel-Cox) test were used for survival analysis. All statistical tests were 2-sided and a p-value < 0.05 was designated as statistically significant. Statistical analysis was performed using GraphPad Prism .The relative expression of Elevated expression levels of circulating miR-21 in breast cancer patientsmiR-21 in breast cancer patients (n=102) was significantly higher , please see MiR-21 expression levels in patients were deregulated with inter-tumoral great variations reflecting tumor heterogeneity and somatic tumor evolution. We used spike-in both Sp6 and MIR-16 to normalize the miR-21 expression. Comparing miR-21 expression in patients younger and older than 40 years-old revealed a significant difference , MiR-21 expression in breast cancer patients with histopathological good/moderate (Grade I-II) and poor (Grade III) differentiation was not significantly different , miR-21 expression in early stage (Stadium I-II) and late stage (Stadium III-IV) was not significantly different , MiR-21 expression levels were also not significantly different according to tumor sizes and breast cancer subtypes , expression levels of circulating subtypes .Expression of miR-21 after surgery and chemotherapymiR-21 expression in breast cancer patients that have completed surgery and chemotherapy was performed in 40 patients. Compared to the expression at diagnosis, miR-21 expression was significantly lower after completion of therapy , Quantification of Correlation of circulating miR-21 upregulation with progression-free survivalmiR-21 expression at diagnosis was determined as an average miR-21 expression in healthy women plus 2xSD as described at the Material and Methods. Progression-free survival was compared between patients with and without miR-21 upregulation at diagnosis. Patients with miR-21 upregulation showed shorter progression-free survival, median survivals were 72 vs 86 weeks and Mantel-Cox test p=0.049, Upregulation of miR-21 as therapeutic monitoring and prognosis biomarkers in breast cancer. Circulating miR-21 expression was significantly lower after surgery and chemotherapy and the upregulation was associated with shorter PFS . However, limited studies have been performed to evaluate the expression levels of circulating miR-21 and the biological correlation with treatment response and survival in breast cancer patietns. To develop novel non-invasive microRNA-based biomarkers in breast cancer, several approaches have been studied including those using plasma or serum  and whole blood . As previously reported that plasma profile is able to detect any alteration after tumor surgery, plasma was used in this study. We investigated the potential role of circulating miR-21 as a minimally-invasive marker for diagnostic and treatment monitoring marker in breast cancer. We presented the potential roles of circulating rter PFS  and 6. AmiR-21 expression as prognostic marker involving more than 1,400 breast cancer patients and both studies have shown that upregulation of miR-21 is correlated with shorter overall survival. Two studies  measured circulating miR-21 expression in a total of 453 breast cancer patients and showed that the upregulation was significantly associated with shorter overall survival. In accordance with previous reports , our study showed that circulating miR-21 expression was significantly higher in breast cancer patients compared to healthy women. We also primarily demonstrated that miR-21 expression decreased significantly after surgery and chemotherapy rendering the potential role as therapeutic monitoring marker. Measuring in breast cancer patient’s whole blood, the previous study did not observe different miR-21 expression measured before and after surgery . Alterations of microRNA expression after surgical resection were also not observed in the whole blood of patients with lung cancer . Different from plasma, whole blood contains both extracellular  and intracellular (within blood cells) microRNAs that might influence to relevance to the presence of the tumor per se. As we found decreased levels of circulating miR-21 after treatment, further study to related the magnitude of the reduction with clinical parameters and outcome. Jinling et al., (2017) and Wang et al., (2015) have performed meta-analysis of miR-21 were not significantly different among different clinical stages and tumor differentiation grades. Other previous studies  also did not find an association of miR-21 expression with baseline stages and differentiation grades. We presented higher expression of circulating miR-21 in young breast cancer patients (less than 40-year-old) that are commonly associated with aggressive biology and poor prognosis . Although further study is required, our finding might reflect that miR-21 upregulation is an important marker for diagnosis and therapeutic response. Several studies have shown the deregulation of miR-21 expression in a subgroup of triple negative breast cancers  that have also been associated with more aggressive behavior and poor prognosis. Other studies have shown an association between miR-21 upregulation (together with deregulation of other circulating microRNAs) with hormonal status , histopathology grades , and distant metastases . We confirmed previous studies  by showing that upregulation of circulating miR-21 at baseline is associated with lower progression-free survival in breast cancer. Although other studies  do not show the correlation with breast cancer prognosis, factors related to tumor heterogeneity, clinical presentation at diagnosis, and overall treatment have also influenced the disease outcome.Expression levels of circulating miR-21 expression after surgery and chemotherapy suggesting the value of a biomarker in response to the decreased tumor burden. However, upcoming study evaluating the magnitude of decreased miR-21 expression and the association with various clinicopathological and clinical outcome of breast cancer patients is required. A constraint of this initial study is the variability of baseline clinical stages, treatment received, potential tumor heterogeneity, and relative small patient cohort. However, we showed that circulating miR-21 expression at diagnosis was not significantly different according to various stages, tumor sizes, tumor differentiation, and breast cancer subtypes. Further study might address circulating miR-21 expression in different breast cancer subtypes, response to specific chemotherapy regimens and hormonal therapy in a bigger / multicenter cohort. The major strength of this initial study is the additional evaluation of circulating All authors have declared for no financial and professional competing interests exist.


The authors identified phosphorylation of ERK and p38MAPK as a surrogate measure to predict better outcome in BRAF-mutant mismatch repair (MMR)-proficient colorectal cancer patients, who were diagnosed with stage III disease. The prognostic value for combined nuclear pERK and p38 score remained significant after multivariate analysis. The authors did not observe this association in MMR-deficient (MMR-D) BRAF-mutant colorectal cancer patients.We read the interesting paper by Roseweir et al.,2 CIMP phenotype is known to be associated with sporadic microsatellite instability-high (MSI-H) colon cancers due to frequent methylation of MLH-1.3 Approximately 60% of loss of MLH-1 expression loss is attributable to BRAF mutations.3 To date, several other studies suggested that more than half of the BRAF-mutant colorectal cancer patients carry the MSI-H phenotype due to loss of MLH-1 function,6 particularly in patients with early stage disease. However, in the paper by Roseweir et al., the frequency of MSI-H tumours in BRAF-mutant stage I−III colorectal cancer patients was only 16% , suggesting this cohort may not represent the general BRAF-mutant colorectal cancer population . Relatively lower incidence of this important subset of patients might have also impacted the multivariate analysis conducted by the authors. Lastly, in the subgroup analysis of colorectal cancer patients, strong nuclear pERK1/2 pattern was significantly  less frequent in patients with MMR-D tumours. The combined nuclear pERK/p38 score was also relatively less detected in MMR-D cases with a borderline statistical significance  suggesting there may be an inverse association between MMR-D status and the pERK/p38 score. Given MMR-D colorectal cancers patients constitute a relatively larger segment of BRAF-mutant patients which is associated with better outcomes due to MSI-H phenotype,4 reported inverse relation between MSI-H and pERK/p38 score may generate further challenges regarding the applicability of this score.BRAF mutations in colorectal cancer are strongly associated with CpG Island methylator phenotype (CIMP), which leads to gene silencing due to methylation of promoter areas of certain genes.Overall, although the authors uncovered a potential important biomarker in BRAF-mutant colorectal cancer patients with MSS tumours, the title of the article does not appear to reflect the authors’ findings, as the association they found was limited only to MMR-proficient BRAF-mutant patients. This constitutes a relatively smaller subset of BRAF-mutant colorectal cancer patients, particularly considering that the association was significant only in stage III tumours. Moreover, these findings may need to be further confirmed in another cohort as the relationship between BRAF mutation and MMR-D reported in this article appears to be weak and is not consistent with other reported literature as discussed above.


Post I-131 ablation single-photon emission computed tomography (SPECT)/CT can show radioactive iodine (RAI)-avid cervical metastatic lymph nodes (mLN) in differentiated thyroid cancer. This study aimed to evaluate the incidence of RAI-avid mLN on post I-131 ablation SPECT/CT and the risk factors related to metastasis among patients with papillary thyroid cancer (PTC) in the low- or intermediate-risk groups.Among 339 patients with PTC who underwent total thyroidectomy followed by I-131 ablation, 292  belonging to the low- or intermediate-risk groups before I-131 ablation, and with sufficient clinical follow-up data were enrolled. The risk groups were classified based on the American Thyroid Association 2015 guideline. Each patient was followed-up for at least 24 months after the ablation (median: 30 months). The clinical, pathologic, and biochemical factors of PTC were reviewed, and their relationships to RAI-avid mLN on SPECT/CT were analyzed.Of the 292 patients, 61 and 231 belonged to the low-and intermediate-risk groups, respectively. Four (6.5%) patients in the low-risk group and 31 (13.0%) patients in the intermediate-risk group had RAI-avid mLN. A high preablation TSH-stimulated serum thyroglobulin (Tg) level in the low- or intermediate-risk group predicted the presence of RAI-avid mLN : 2.96; p = 0.04). In the subgroup analysis by risk group, TSH-stimulated serum Tg only predicted RAI-avid mLN in the low-risk group .The incidence of RAI-avid mLN on postablation SPECT/CT was relatively high in both low- and intermediate-risk patients with PTC, and high preablation TSH-stimulated serum Tg level was a predictor of metastasis, especially in the low-risk group. A selective treatment approach should be considered in patients with high preablation TSH-stimulated serum Tg level. Residual normal or malignant thyroid tissue after total thyroidectomy for differentiated thyroid carcinoma (DTC) is a common finding. Radioactive iodine (RAI) can be administered to thyroidectomized patients for remnant ablation or as an adjuvant therapy. Some retrospective studies have shown that RAI treatment improves the survival rate and decreases the recurrence rate of papillary thyroid cancer (PTC) , 2.However, the 2015 American Thyroid Association (ATA) Management Guidelines for DTC do not recommend routine RAI remnant ablation for low-risk DTC patients . The incThe introduction of single-photon emission computed tomography (SPECT)/CT technology has increased the diagnostic value of RAI scintigraphy. SPECT/CT can detect LN metastasis more sensitively and more specifically than a planar scan –6 and caThe current study aimed to evaluate the incidence of hidden RAI-avid mLN on RxSPECT/CT after curative surgery in patients with PTC belonging to the low- or intermediate-risk groups according to the 2015 ATA guideline, and to investigate the risk factors related to the occurrence of metastasis.This study involved 339 patients with PTC who underwent total thyroidectomy followed by RAI ablation in our hospital between September 2013 and December 2014. The patients who belonged to the low- or intermediate-risk group before RAI ablation were enrolled [The survey was approved by the institutional review board (IRB) of Kyungpook National University School of Medicine/Hospital, Korea. The patient records and information was anonymized and de-identified prior to analysis and all data was fully anonymized prior to access by any of the authors.RAI ablation was performed at a median interval of 7 weeks (range: 4–14 weeks) after operation, either after the injection of recombinant human TSH  or after the withdrawal of thyroid hormone replacement (THW). Depending on the pathological result, the administered doses of I-131 ranged from 1.11 GBq to 5.55 GBq (30–150 mCi), and postablation I-131 scintigraphy was performed 4 to 5 days after treatment using a dual-head gamma camera equipped with 1.5875 cm NaI crystals and a multidetector (16-row) spiral CT . Post RAI ablation whole-body planar scan (RxWBS) was initially acquired at a scanning speed of 10 cm/min, after which SPECT/CT was performed. A 128 × 128 matrix was used, and 6430-sec projections were acquired over 360 degrees for the SPECT acquisition. SPECT data were reconstructed using a 3-dimensional iterative algorithm. CT scans were obtained at a tube voltage of 120 kV. The slice width and spacing were 3.75 mm. CT data were used for anatomical information and attenuation correction. Data of RxWBS and SPECT/CT were analyzed with Xeleris software . No contrast medium was used during the procedure.We measured serum TSH, Tg, and TgAb immediately before RAI administration. The serum Tg and serum TgAb levels were evaluated using Thyroglobulin IRMA  and Tg Antibody Coated Tube Assay  kits. Meanwhile, serum TSH  was measured via immunoradiometric assay commercial kits. TSH levels > 30 μU/mL were considered adequate to increase RAI uptake in thyroid tumor cells during RAI ablation.All images from RxWBS and RxSPECT/CT were analyzed separately by two experienced nuclear medicine physicians. On RxWBS, RAI uptake in the esophageal tract and salivary glands were considered to be physiological, and other focal tracer uptake on the background that was not compatible with the residual thyroid tissue or physiological activity was considered as a pathologic uptake. The SPECT/CT images were classified as (a) remnant tissue if the median foci were localized either in the upper thyroid bed, in the thyroglossal duct remnants, or in the lower thyroid bed or (b) LN metastasis if the focus was localized in front of a round lesion identical to an LN.U-test. To identify the optimal cut-off for serum Tg level for predicting RAI-avid mLN on RxSPECT/CT, receiver operating characteristic (ROC) curve analysis was used. Logistic regression was used for the multivariate analysis. MedCalc Statistical Software version 13.2 was used for all statistical calculations, and p values < 0.05 were considered statistically significant.Continuous data are presented as medians and ranges or means and standard deviations, as appropriate for each variable. Chi-square tests were performed for categorical comparisons, and continuous variables were compared using Mann–Whitney A total of 292 patients  belonged to the low- (N = 61) or intermediate-risk (N = 231) groups before RAI ablation. The demographic characteristics of the patients are summarized in A RAI-avid mLN was identified on RxSPECT/CT in 35 (12%) of the patients . The cliFour of the 61 patients (6.5%) in the low-risk group showed RAI-avid mLN and were recategorized into the intermediate-risk group due to the presence of mLN. Hidden RAI-avid mLN was predicted only by preablation TSH-stimulated serum Tg among patients in the low-risk group . The optIn the intermediate-risk group (N = 231 patients), 31 patients (13.0%) had hidden RAI-avid mLN on RxSPECT/CT. Patients with RAI-avid mLN had higher TSH-stimulated serum Tg values than patients without mLN, but the difference was not statistically significant. All clinicopathological data according to mLN status on RxSPECT/CT were not significantly different in this group .In our study, the incidence of RAI-avid mLN that was newly identified on RxSPECT/CT was relatively high  in patients with PTC who belonged to the low- or intermediate-risk groups. Four (6.5%) out of the 61 patients in the low-risk group had postoperative hidden RAI-avid mLN, and their risk status was changed to intermediate after the identification of the mLN. In this study, all subjects received the prophylactic central neck dissection, therefore this result might not be similar to patients who did not receive prophylactic central neck dissection. We think that frequency of LN metastasis could be higher in patients did not received the prophylactic central neck dissection.Among the clinical and pathological parameters, preablation TSH-stimulated serum Tg level was the sole predictor of the presence of hidden RAI-avid mLN. In the subgroup analysis, preablation TSH-stimulated serum Tg level was the only predictor of the presence of RAI-avid mLN in the low-risk group. However, no significant predictors of RAI-avid mLN were found in the intermediate-risk group.Many retrospective studies have reported that postoperative TSH-stimulated serum Tg is a useful predictor of persistent/recurrent disease in thyroidectomized patients with thyroid cancer –11. SomeThe optimal cut-off value of TSH-stimulated serum Tg for predicting RAI-avid mLN as determined via ROC curve analysis was 0.5 ng/ml in low or intermediate-risk group . However only 11 (9.6%) among 114 patients having TSH-stimulated serum Tg levels < 0.5 ng/ml had RAI-avid mLN . In low-Before the introduction of SPECT/CT technology, planar post I-131 ablation scan had been used to identify RAI-avid metastatic foci. However, differentiating RAI-avid mLN from physiological RAI accumulation and remnant thyroid tissue on planar post I-131 ablation scan might be difficult due to the lack of anatomical detail and the overlap of RAI uptake areas. The combination of SPECT/CT and planar scan provides additional information about RAI-avid metastatic foci –6, 16. WOur study had several limitations. First was the retrospective design of our study in which we tried to include patients who underwent surgery and followed up in one hospital to ensure a similar therapeutic approach, imaging studies, and laboratory evaluation. Second, although the low-risk group defined by the 2015 ATA guideline includes clinical N0 or under 5 pathologic N1 micrometastases (< 0.2 cm in the largest dimension), we had limited information regarding the size of mLN. Each patient in the low-risk group is considered to be in the N0 clinical stage, which means that no suspicious LN metastasis was found on preoperative neck US. Even though cervical mLN might be present, it would be too small to be detected on preoperative US, which might mean no suspicious macrometastatic LN. Third, RAI was administered either after rhTSH or after THW, and the TSH-stimulated serum Tg levels might be affected by methods for TSH elevation . HoweverIn the current study, patient risk was stratified according to the 2015 ATA guideline, and RAI-avid mLN was assessed by SPECT/CT. Our results demonstrated that preablation TSH-stimulated serum Tg can predict hidden RAI-avid mLN, especially in the low-risk group.The incidence of RAI-avid mLN on RxSPECT/CT was relatively high in both low- and intermediate-risk patients with PTC, and high preablation TSH-stimulated serum Tg level was a predictor of metastasis, especially in the low-risk group. The results of the current study suggest that the therapeutic plan, including RAI ablation, can be individualized according to preablation TSH-stimulated serum Tg level in low-risk patients with PTC.