BACKGROUND

Medulloblastoma (MB) is a rare primary brain tumor in adults. We previously evaluated that combining both clinical and molecular classification could improve current risk stratification for adult MB. In this study, we aimed to identify the prognostic value of Ki-67 index in adult MB.

METHODS

Ki-67 index of 51 primary adult MBs was reassessed using a computer-based image analysis (Image-Pro Plus). All patients were followed up ranging from <em>12</em> months up to 15 years. Gene expression profiling and immunochemistry were used to establish the molecular subgroups in adult MB. Combined risk stratification models were designed based on clinical characteristics, molecular classification and Ki-67 index, and identified by multivariable Cox proportional hazards analysis.

RESULTS

In our cohort, the mean Ki-67 value was 30.0 ± 11.3% (range 6.56-63.55%). The average Ki-67 value was significantly higher in LC/AMB than in CMB and DNMB (P = .001). Among three molecular subgroups, Group 4-tumors had the highest average Ki-67 value compared with WNT- and SHH-tumors (P = .004). Patients with Ki-67 index large than 30% displayed poorer overall survival (OS) and progression free survival (PFS) than those with Ki-67 less than 30% (OS: P = .001; PFS: P = .006). Ki-67 index (i.e.>> 30%, < 30%) was identified as an independent significant prognostic factor (OS: P = .017; PFS: P = .024) by using multivariate Cox proportional hazards model.

CONCLUSIONS

In conclusion, Ki-67 index can be considered as a valuable independent prognostic biomarker for adult patients with MB.

Hyperactive Wnt signaling is frequently observed in colorectal cancer. Higher intakes of dietary fiber [nondigestible carbohydrates (NDCs)] and the fermentation product butyrate are protective against colorectal cancer and may exert their preventative effects via modulation of the Wnt pathway.

We investigated the effects of supplementing healthy individuals with 2 NDCs [resistant starch (RS) and polydextrose] on fecal calprotectin concentrations and Wnt pathway-related gene expression. In addition, we determined whether effects on secreted frizzled-related protein 1 (SFRP1) expression are mediated via the epigenetic mechanisms DNA methylation and microRNA expression.

In a randomized, double-blind, placebo-controlled trial (the Dietary Intervention, Stem cells and Colorectal Cancer (DISC) Study), 75 healthy participants were supplemented with RS and/or polydextrose or placebo for 50 d in a 2 × 2 factorial design. Pre- and postintervention stool samples and rectal mucosal biopsies were collected and used to quantify calprotectin and expression of 12 Wnt-related genes, respectively. The expression of 10 microRNAs predicted to target SFRP1 was also quantified by quantitative reverse transcriptase-polymerase chain reaction, and DNA methylation was quantified at 7 CpG sites within the SFRP1 promoter region by pyrosequencing.

NDC supplementation did not affect fecal calprotectin concentration. SFRP1 mRNA expression was reduced by both RS (P = 0.005) and polydextrose (P = 0.053). RS and polydextrose did not affect SFRP1 methylation or alter the expression of 10 microRNAs predicted to target SFRP1. There were no significant interactions between RS and polydextrose.

RS and polydextrose supplementation did not affect fecal calprotectin concentrations. Downregulation of SFRP1 with RS and polydextrose could result in increased Wnt pathway activity. However, effects on Wnt pathway activity and downstream functional effects in the healthy large-bowel mucosa remain to be investigated. The DISC Study was registered at clinicaltrials.gov as NCT01214681.

Lycorine is a benzylphenethylamine-type alkaloid member of the Amaryllidaceae family. A lycorine derivative, HLY78, was previously identified as a new <em>Wnt</em>/β-catenin signaling pathway agonist that targets the DAX domain of axin. Herein, the structural optimization of HLY78 and analyses of the structure-activity relationships of lycorine-derived phenanthridine derivatives as agonists of the <em>Wnt</em>/β-catenin signaling pathway are presented. This research suggests that triazole groups are important pharmacophores for <em>Wnt</em> activation; triazole groups at C-8 and C-9 of phenanthridine compounds markedly enhanced <em>Wnt</em> activation. A C-11-C-<em>12</em> single bond is also important for <em>Wnt</em> activation. On the basis of these findings, two <em>Wnt</em> agonists were designed and synthesized. The results for these agonists indicated that the combination of a 4-ethyldihydrophenanthridine skeleton and a triazole substituent improves <em>Wnt</em> activation. These compounds may be useful in further pharmacological or biological studies.

This study investigates the effects of aspirin (ASA) on the proliferative capacity, osteogenic potential, and expression of growth factor-associated genes in periodontal ligament stem cells (PDLSCs).

Mesenchymal stem cells (MSCs) from PDL tissue were isolated from human premolars (n = 3). The MSCs' identity was confirmed by immunophenotyping and trilineage differentiation assays. Cell proliferation activity was assessed through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Polymerase chain reaction array was used to profile the expression of 84 growth factor-associated genes. Pathway analysis was used to identify the biologic functions and canonic pathways activated by ASA treatment. The osteogenic potential was evaluated through mineralization assay.

ASA at 1,000 μM enhances osteogenic potential of PDLSCs. Using a fold change (FC) of 2.0 as a threshold value, the gene expression analyses indicated that 19 genes were differentially expressed, which includes <em>12</em> upregulated and seven downregulated genes. Fibroblast growth factor 9 (FGF9), vascular endothelial growth factor A (VEGFA), interleukin-2, bone morphogenetic protein-10, VEGFC, and 2 (FGF2) were markedly upregulated (FC range, 6 to 15), whereas pleotropin, FGF5, brain-derived neurotrophic factor, and Dickkopf <em>WNT</em> signaling pathway inhibitor 1 were markedly downregulated (FC 32). Of the 84 growth factor-associated genes screened, 35 showed high cycle threshold values (≥35).

ASA modulates the expression of growth factor-associated genes and enhances osteogenic potential in PDLSCs. ASA upregulated the expression of genes that could activate biologic functions and canonic pathways related to cell proliferation, human embryonic stem cell pluripotency, tissue regeneration, and differentiation. These findings suggest that ASA enhances PDLSC function and may be useful in regenerative dentistry applications, particularly in the areas of periodontal health and regeneration.

<strong class="sub-title"> Introduction: </strong> Chronic myeloid leukemia (CML) is a myeloid malignancy characterized by the oncogene BCR-ABL. CML responds well to therapy targeting BCR-ABL in the chronic phase but is resistant to treatment when it progresses to the blast phase (BP). This study attempted to address whether arachidonate <em>12</em>-lipoxygenase (Alox<em>12</em>) confers to CML drug resistance.

<strong class="sub-title"> Materials and methods: </strong> We analyzed the expression of Alox<em>12</em> using Western blotting, ELISA, and RT-PCR methods. Loss of functional analysis was performed using cellular activity assays on CML and normal hematopoietic stem/progenitor cells (HSPCs).

<strong class="sub-title"> Results: </strong> Alox<em>12</em> and <em>12</em>-Hydroxyeicosatetraenoic acid (<em>12</em>-HETE) are overexpressed in BP-CML but not HSPCs, and that Alox<em>12</em>-<em>12</em>-HETE axis is regulated by BCR-ABL. The Alox<em>12</em>-<em>12</em>-HETE axis is required for CML. Specific Alox<em>12</em> inhibitor inhibits colony formation, survival, and self-renewal capacity in BP-CML HSPCs, and to a significantly greater extent than in normal HSPCs. Of note, the Alox<em>12</em> inhibitor significantly augments dasatinib's efficacy in BP-CML HSPCs. Mechanism studies show that Alox<em>12</em> inhibition does not affect activities of essential signaling pathways involved in maintaining stem cell function, such as Wnt, p53, and bone morphogenetic protein (BMP). In contrast, we show that Alox<em>12</em> inhibition disrupts nicotinamide adenine dinucleotide phosphate (NADPH) homeostasis and induces oxidative stress and damage in CML HSPCs and committed cells.

<strong class="sub-title"> Conclusion: </strong> Alox<em>12</em>-<em>12</em>-HETE axis is a specific and critical regulator of BP-CML HSPCs functions. Pharmacological inhibition of Alox<em>12</em> may be useful in BP-CML.

<strong class="sub-title"> Keywords: </strong> Alox<em>12</em>; Bcr-Abl; chronic myeloid leukemia; resistance; stem cell.

Nucleotide-activated effector deployment, prototyped by interferon-dependent immunity, is a common mechanistic theme shared by immune systems of several animals and prokaryotes. Prokaryotic versions include CRISPR/Cas with the CRISPR polymerase domain, their minimal variants, and systems with Second Messenger Oligonucleotide or Dinucleotide Synthetase (SMODS). Cyclic or linear oligonucleotide signals in these systems help set a threshold for the activation of potentially deleterious downstream effectors in response to invader-detection. We establish such a regulatory mechanism to be a more general principle of immune systems, which can also operate independently of such messengers. Using sensitive sequence analysis and comparative genomics we identify <em>12</em> new prokaryotic immune systems, which we unify by this principle of threshold-dependent effector activation. These display regulatory mechanisms paralleling physiological signaling based on 3'-5' cyclic mononucleotides, NAD⁺-derived messengers, two- and one-component signaling that include histidine kinase-based signaling, and proteolytic activation. Further, these systems allowed identification of multiple new sensory signal sensory components, such as a tetratricopeptide-repeat-scaffold predicted to recognize NAD⁺-derived signals, unreported versions of the STING domain, prokaryotic YEATS domains, and a predicted nucleotide-sensor related to receiver domains. We also identify previously unrecognized invader-detection components and effector components, such as prokaryotic versions of the <em>Wnt</em> domain. Finally, we show that there have been multiple acquisitions of unidentified STING domains in eukaryotes, while the TPR scaffold was incorporated into the animal immunity/apoptosis ASK signalosome.<b>IMPORTANCE</b> Both prokaryotic and eukaryotic immune systems face the danger of premature activation of effectors and degradation of self-molecules in the absence of an invader. To mitigate this, they have evolved threshold-setting regulatory mechanisms for the triggering of effectors only upon detection of a sufficiently strong invader-signal. This work defines general templates for such regulation in effector-based immune systems. Using this, we identify several previously uncharacterized prokaryotic immune mechanisms that accomplish regulation of downstream effector deployment by using nucleotide, NAD⁺-derived, two-component and one-component signals paralleling physiological homeostasis. This study has also helped identify several previously unknown sensor and effector modules in these systems. Our findings also augment the growing evidence for the emergence of key animal immunity and chromatin regulatory components from prokaryotic progenitors.

<AbstractText>Moniezia expansa (Cyclophyllidea: Anoplocephalidae) is a large species of tapeworm that occurs in sheep and cattle and inhabits the small intestine, causing diarrhea and weight declines, leading to stockbreeding losses. Interestingly, the body fat percentage of M. expansa, which lacks the ability to synthesize fatty acids, is as high as 78% (dry weight) and all of the proglottids of M. expansa exhibit a dynamic developmental process from top to bottom. The aim of this paper is to identify the molecular basis of this high body fat percentage, the dynamic expression of developmental genes and their expression regulation patterns.</AbstractText><AbstractText>From <em>12</em> different proglottids (four sections: scolex and neck, immature, mature and gravid with three replicates), 13,874 transcripts and 680 differentially expressed genes (DEGs) were obtained. The gene expression patterns of the scolex and neck and immature proglottids were very similar, while those of the mature and gravid proglottids differed greatly. In addition, 13 lipid transport-related proteins were found in the DEGs, and the expression levels showed an increasing trend in the four proglottid types. Furthermore, it was shown that 33 homeobox genes, 9 of which were DEGs, had the highest expression in the scolex and neck section. The functional enrichment results of the DEGs were predominantly indicative of development-related processes, and there were also some signal transduction and metabolism results. The most striking result was the finding of <em>Wnt</em> signaling pathways, which appeared multiple times. Furthermore, the weighted gene co-expression networks were divided into <em>12</em> modules, of which the brown module was enriched with many development-related genes.</AbstractText><AbstractText>We hypothesize that M. expansa uses lipid transport-associated proteins to transport lipids from the host gut to obtain energy to facilitate its high fecundity. In addition, homeobox genes and <em>Wnt</em> signaling pathways play a core role in development and regeneration. The results promote research on the cell differentiation involved in the continuous growth and extension of body structures.</AbstractText>

<AbstractText>Adipose-derived stromal/stem cells (ASCs) have attracted attention as a promising material for regenerative medicine. Previously, we reported an age-related decrease in the adipogenic potential of ASCs from human subjects and found that the individual difference in this potential increased with age, although the mechanisms remain unclear. Recently, other groups demonstrated that a secreted antagonist of bone morphogenetic protein (BMP) signaling, Gremlin 2 (GREM2), inhibits the differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) into osteoblasts and the adipogenesis of 3T3-L1 cell. Here, we examined the effects of GREM2 on the differentiation of ASCs into adipocytes.</AbstractText><AbstractText>To examine changes in GREM2 expression levels with age, immunohistochemistry was performed on subcutaneous adipose tissues from subjects <em>12</em>-97 years of age. Next, GREM2 gene expression levels in ASCs collected from subjects 5-90 years of age were examined by RT-PCR, and the change with age and correlation between the expression level and the adipogenic potential of ASCs were analyzed. In addition, to assess whether GREM2 affects adipogenesis, ASCs (purchased from a vendor) were cultured to induce adipogenesis with recombinant GREM2 protein, and siRNA-induced GREM2 knockdown experiment was also performed using aged ASCs.</AbstractText><AbstractText>In adipose tissues, GREM2 expression was observed in cells, including ASCs, but not in mature adipocytes, and the expression level per cell increased with age. GREM2 expression levels in ASCs cultured in vitro also increased with age, and the individual differences in the level increased with age. Of note, partial correlation analysis controlled for age revealed that the adipogenic potential of ASCs and the GREM2 gene expression level were negatively correlated. Furthermore, based on a GREM2 addition experiment, GREM2 has inhibitory effects on the adipogenesis of ASCs through activation of <em>Wnt</em>/β-catenin signaling. On the other hand, GREM2 knockdown in aged ASCs promoted adipogenesis.</AbstractText><AbstractText>The GREM2 expression level was confirmed to play a role in the age-related decrease in adipogenic potential observed in ASCs isolated from adipose tissues as well as in the enhancement of the individual difference, which increased with age. GREM2 in adipose tissues increased with age, which suggested that GREM2 functions as an inhibitory factor of adipogenesis in ASCs.</AbstractText>

SRY-related high-mobility-group box <em>12</em> (SOX<em>12</em>) has currently emerged as a key cancer-related protein in multiple human cancer types. However, little is known about the relevance of SOX<em>12</em> in multiple myeloma (MM). The current study aimed to investigate the potential role of SOX<em>12</em> in MM. Our results demonstrated that SOX<em>12</em> expression was markedly elevated in MM cell lines. A series of cellular assays demonstrated that SOX<em>12</em> knockdown significantly reduced the proliferation and colony formation, and upregulated cell apoptosis of MM cells. By contrast, SOX<em>12</em> overexpression promoted the proliferation, colony formation and decreased the apoptosis of MM cells, results that reveal its oncogenic effects. SOX<em>12</em> regulated β-catenin expression and TCF/LEF transcriptional activity. Moreover, the SOX<em>12</em>-knockdown-mediated antitumor effect in MM cells was significantly reversed by transfecting a β-catenin expression vector. Notably, SOX<em>12</em> inhibition retarded tumor growth in vivo of a MM-derived mouse xenograft model. In conclusion, our results suggest a potential oncogenic function for SOX<em>12</em> in MM. Our findings reveal that SOX<em>12</em> knockdown inhibits the growth of MM cells by downregulating the <em>Wnt</em>/β-catenin signaling pathway, results that imply SOX<em>12</em> may represent a novel therapeutic target for MM treatment.

We reported recently that berberine, a traditional oriental medicine to treat gastroenteritis, binds and activates Retinoid X receptor α (RXRα) to suppress the growth of colon cancer cells. Here, we extended our studies based on the binding mode of berberine with RXRα by design, synthesis and biological evaluation of a focused library of 15 novel berberine analogues. Among them, 3,9-dimethoxy-5,6-dihydroisoquinolino[3,2-a]isoquinolin-7-ium chloride (<b>B-<em>12</em></b>) was identified as the optimal RXRα activator. More efficiently than berberine, <b>B-<em>12</em></b> bound and altered the conformation of RXRα/LBD, thereby suppressing <em>Wnt</em>/β-catenin pathway and colon cancer cell growth via RXRα mediation. In addition, <b>B-<em>12</em></b> not only preserved berberine's tumor selectivity but also greatly improved its bioavailability. Remarkably, in mice, <b>B-<em>12</em></b> did not show obvious side effects including hypertriglyceridemia as other RXRα agonists, or induce hepatorenal toxicity. Together, our study describes an approach for the rational design of berberine-derived RXRα activators as novel effective antineoplastic agents for colon cancer.

OBJECTIVE

To investigate the serum protein targets of Qianggu Decoction (, QGD) on treating osteoporosis by the proteomics analysis using tandem mass tag (TMT) and liquid chromatographytandem mass spectrometry (LC-MS/MS).

METHODS

Twenty serum protein samples were recruited (10 patients with primary type I osteoporosis before and after QGD treatment) and the high abundance ratios protein was removed, two serum samples were extracted and labeled with TMT reagent. Then, mass spectrometric detection, identification of differentially expressed proteins and bioinformatics analysis of differentially expressed proteins were carried out.

RESULTS

A total of 60 proteins were identified, within a 99% confidence interval, to be differentially regulated of which, 34 proteins were up-regulated and 26 proteins were down-regulated. Differentially expressed proteins analyzed by Gene Ontology (GO) annotation mainly get involved in <em>12</em> different biological processes, 7 types of cellular components, and 6 kinds of molecular functions. Angiotensinogen (AGT), stromelysin-1 (MMP3), heparanase (HPSE) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were screened as candidate protein targets of QGD treatment, which were related to metabolic mechanism of bone remodeling and/or bone collagen of osteoporosis. By the utilization of the protein-protein interaction network analysis tool named STRING10.0, it showed that AGT, MMP3, HPSE and GAPDH were located in the key node of the protein-protein interactions network. Furthermore, AGT, MMP3, HPSE and GAPDH were found to be directly related to BMP, MAPK, <em>Wnt</em>, SMAD and tumor necrosis factor ligand superfamily member 11 (TNFSF11) families.

CONCLUSIONS

The proteomics analysis by using TMT combined with LC-MS/MS was a feasible method for screening the potential therapeutic targets associated with QGD treatment. It suggests that AGT, MMP3, HPSE and GAPDH may be candidate protein targets of QGD treatment which can be used as therapeutic effect monitor and early diagnosis of primary type I osteoporosis.

Lithium activates <em>Wnt</em>/β-catenin signaling leading to stabilization of free cytosolic β-catenin. The aim of the present study is to evaluate the in vivo effect of acute and chronic lithium treatment on the expression of β-catenin target genes, addressing its transcripts HIG2, Bcl-xL, Cyclin D1, c-myc, in cortical and hippocampal tissue from adult mice. Lithium doses were established to yield therapeutic working concentrations. In acute treatment, mice received a 300µL of a 350 mg/kg solution of LiCl by gavage, and were euthanized after 2 h, 6 h and <em>12</em> h. To determine the effect of chronic treatment, animals were continuously fed either with chow supplemented with 2 g/kg Li2CO3, or regular chow (controls), being euthanized after 30 days. All animals had access to drinking water and 0.9% saline ad libitum. After acute and chronic treatments samples of peripheral blood were obtained from the tail vein for each animal, and serum concentrations of lithium were determined. All transcripts were up-regulated in cortical and hippocampal tissues of lithium-treated mice, both under acute and chronic treatments. There was a positive correlation between serum lithium concentrations and the increment in the expression of all transcripts. This effect was observed in all time points of the acute treatment (i.e., 2, 6 and <em>12</em> hours) and also after 30 days. We conclude that <em>Wnt</em>/β-catenin transcriptional response (HIG2, Bcl-xL, Cyclin D1 and c-myc) is up-regulated in the mouse brain in response to acute and chronic lithium treatment at therapeutic concentrations.

Keywords: Bcl-xL; Cyclin D1; HIG2; Lithium; Wnt/β-catenin; c-myc.

<strong class="sub-title"> Introduction: </strong> Established thresholds for patient-reported outcomes (PROs) provide clinically relevant responder data from trials. Lorecivivint (LOR) is an intra-articular (IA) therapy in development for knee osteoarthritis (OA). A post hoc analysis from a phase 2b trial (<a href="http://clinicaltrials.gov/show/NCT03<em>12</em>2860" title="See in ClinicalTrials.gov">NCT03<em>12</em>2860</a>) determined proportions of LOR responders.

<strong class="sub-title"> Methods: </strong> A 24-week, randomized trial of 0.07 mg LOR demonstrated PRO improvements compared with PBO in moderate-to-severe knee OA participants. Participants treated with LOR and PBO achieving 30%/50%/70% improvements at weeks <em>12</em> and 24 in Pain Numeric Rating Scale (NRS), WOMAC Pain/Function subscales, Patient Global Assessment (PtGA), and OMERACT-OARSI responder criteria were determined. Odds ratios (ORs) and 95% confidence intervals [CIs] were compared with PBO.

<strong class="sub-title"> Results: </strong> There were 115 and 116 participants in the LOR and PBO groups, respectively. For Pain NRS, LOR increased ORs of achieving 30% [week <em>12</em>, OR = 2.47 (1.45, 4.19), P < 0.001; week 24, OR = 2.37 (1.40, 4.02), P < 0.01] and 50% [week 24, OR = 1.89 (1.11, 3.23), P < 0.05] improvements over baseline. For WOMAC Pain, LOR increased ORs of achieving 30% [week 24, OR = 1.79 (1.06, 3.01), P < 0.05] and 50% [week <em>12</em>, OR = 1.79 (1.06, 3.03), P < 0.05; week 24, OR = 1.73 (1.02, 2.93), P < 0.05] improvements. For WOMAC Function, LOR increased ORs of achieving 30% [week <em>12</em>, OR = 1.85 (1.10, 3.<em>12</em>), P < 0.05; week 24, OR = 1.93 (1.14, 3.26), P < 0.05] improvements. For PtGA, LOR increased ORs of achieving 50% [week <em>12</em>, OR = 2.28 (1.25, 4.16), P < 0.01] improvements. LOR produced numerical increases at the 70% threshold. LOR increased ORs of achieving OMERACT-OARSI responses [week <em>12</em>, OR = 2.21 (1.29, 3.78); P < 0.01; week 24, OR = 2.57 (1.49, 4.43), P < 0.001] and strict responses [week <em>12</em>, OR = 2.13 (1.26, 3.61), P < 0.01; week 24, OR = 2.05 (1.21, 3.47), P < 0.01].

Conclusions: LOR (0.07 mg) demonstrated improved PRO threshold responses across single and composite measures of pain, function, and patient global assessment compared with PBO, with benefits sustained to 24 weeks.

Keywords: Alternative splicing; CLK2; DYRK1A; OMERACT-OARSI; Patient-reported outcomes; Wnt signaling pathway.

<AbstractText>The incidence of metabolic syndrome (MetS) in menopausal women is one of the main health care concerns. MetS clusters are related to an imbalance in pro- and anti-inflammatory adipokines such as secreted frizzled-related protein 5 (SFRP5) and wingless-type mammary tumor virus integration site family, member 5A (<em>WNT</em>5A). <em>WNT</em>5A induces an inflammatory state to induce insulin resistance and further pathologic consequences. Recent strategies to prevent progression of MetS to diabetes have focused on conservative treatments such as exercise and herbal medicine. The aim of this study was to investigate the mechanistic effects of cotreatment with cinnamon extract and <em>12</em>-wk high-intensity endurance training on MetS components considering the non-canonical <em>WNT</em>5A signaling.</AbstractText><AbstractText>Thirty-two female ovariectomized Wistar rats were divided into the following four groups (n = 8/group): exercise (Ova+Exe), cinnamon extract (Ova+Cin), exercise with cinnamon extract (Ova+Exe+Cin) and saline (Ova+Sal). One group of rats undergoing surgery without removal of the ovaries was considered as a sham. After 3 mo of experimental intervention, waist circumference, serum concentrations of glucose, insulin, lipid profile, tumor necrosis factor-α, <em>WNT</em>5A, and SFRP5 were measured.</AbstractText><AbstractText>Data showed a significant reduction in serum glucose, low-density lipoprotein, homeostasis model assessment estimate of insulin resistance, and tumor necrosis factor-α, but an increase in SFRP5 level in Ova+Exe, Ova+Cin and Ova +Exe+Cin groups compared with Ova+Sal group (P < 0.05). Serum <em>WNT</em>5A significantly was reduced only in Ova+Exe+Cin group (P = 0.02).</AbstractText><AbstractText>The present study indicated that high-endurance training combined with aqueous cinnamon extract supplementation for <em>12</em> wk more efficiently alleviated insulin resistance and metabolic dysfunctions via reduction in noncanonical <em>WNT</em> signaling in ovariectomized rats.</AbstractText>

Focal dermal hypoplasia (FDH) or Goltz Syndrome (OMIM# 305600) is an X-linked dominant ectodermal dysplasia caused by mutations in the PORCN gene. This gene encodes an endoplasmic reticulum transmembrane protein that is involved in processing the embryonically critical <em>WNT</em> signaling proteins. Individuals diagnosed with FDH were recruited to participate in the study through the National Foundation for Ectodermal Dysplasia. Individuals were evaluated to characterize the FDH phenotype. Each participant completed a brief dental survey and oral evaluation using artificial light. To identify the oral soft and hard tissue findings 19 individuals (16 female and 3 male) participated with a median age of 10 years (range 2-56 years). Soft and hard tissue defects were present in 68% (13) and 94% (18) of the patients, respectively. Dental anomalies were highly prevalent with 68% (13) demonstrating vertical enamel grooving, 52% (10) having peg shaped tooth deformities, and 78% (15) having enamel hypoplasia with or without discoloration. Cleft lip and cleft palate presented in 15% (3) of the participants. Other findings included 57% (11) having intra-oral lipoma or papilloma with no site predilection. Dental malocclusions were common with 63% (<em>12</em>) having some degree of malocclusion with 15% (3) of participants having class III malocclusion with an anterior dental cross bite. Participants frequently reported speech problems or difficulty with chewing (73%; N = 14). This study shows there is marked variation in the oral phenotype of individuals with FDH and underscores the important role of <em>WNT</em> signaling in oro-facial development.

Titanium dioxide (TiO₂) and nano-sized titanium dioxide (nano-TiO₂), which are used in food production, may be harmful to the body. Long-term exposure to nano-TiO₂ can lead to hepatic injury; however, the effect of nano-TiO₂ on liver fibrosis and the underlying mechanism remain unclear. The TGF-<i>β</i>/Smad/MAPK/<em>Wnt</em> signaling pathway is important for tissue fibrosis. In this study, mice were fed nano-TiO₂ (2.5, 5, and 10 mg/kg body weight) for nine consecutive months to investigate its effect on liver fibrosis. Nano-TiO₂ induced hepatic inflammatory cell infiltration and hepatic fibrosis and upregulated the expression of HIF-1<i>α</i> (+75-fold to +2.38-fold), <em>Wnt</em>3 (+<em>12</em>% to +135%), <em>Wnt</em>4 (1.33-fold to 6-fold), NF-<i>κ</i>B (+3.13% to +34.38%), TGF-<i>β</i>1 (+1307-fold to +1.85-fold), TGF-<i>β</i>1R (+0.8-fold to 1.33-fold), Smad-2 (+0.58-fold to +1.58-fold), ILK (+0.43-fold to +1.19-fold), ECM (+1.82-fold to 2.36-fold), calpain 2 (+0.11-fold to +0.78-fold), <i>α</i>-SMA (+0.63-fold to +1.56-fold), c-Myc (+0.27-fold to +0.46-fold), and collagen I (+8% to +36%), and increased the phosphorylation level of p38MAPK (+66.67% to +153.33%) in inflammatory and fibrotic liver tissues, whereas it downregulated cyclin D (-6.25% to -43.75%) and decreased the phosphorylation levels of GSK-3<i>β</i> (-3.<em>12</em>% to -46.88%) and <i>β</i>-catenin (-19.57% to -45.65%). These results indicate that hepatic fibrosis induced by nano-TiO₂ is mediated by the TGF-<i>β</i>/Smads/MAPK/<em>Wnt</em> signaling pathway. This study provides insight into the mechanism underlying hepatic toxicity induced by nano-TiO₂ .

In previous research, several <em>WNT</em> signaling pathway genes including transcription factor <em>12</em> (TCF<em>12</em>), catenin alpha-like protein 1 (CTNNAL1) and wingless-type MMTV integration site family, member 10B (<em>WNT</em>10B) were differentially expressed in PMSG-hCG stimulated preovulatory ovarian follicles of Large White and Chinese Taihu sows. In the present research, these three genes were selected as the candidate genes for litter size traits in pigs. Four mutations (TCF<em>12</em> c.-201+65 G>A, TCF<em>12</em> c.-200-300 G>A, CTNNAL1 c.1878 G>C and <em>WNT</em>10B c.*<em>12</em> C>T) were detected in eleven pig populations, and results indicated CTNNAL1 c.1878 G and <em>WNT</em>10B c.*<em>12</em> C were the major alleles in all tested pig populations, while TCF<em>12</em> c.-201+65 A and TCF<em>12</em> c.-200-300 A were the major alleles in several Chinese native pig breeds. Association analysis of four mutations with litter size in Large White and DIV pigs showed that both the signficant differences of total number born (TNB) and number born alive (NBA) among three genotypes and the significance of additive effects appeared at TCF<em>12</em> c.-200-300 G>A and CTNNAL1 c.1878 G>C loci, suggesting these two mutations might be reliable markers for pig selection and breeding.

<AbstractText>Medulloblastomas may occur in a predisposition context, including familial adenomatosis polyposis. Medulloblastomas related to APC germline pathogenic variant remain rare and poorly described. Their similarities with sporadic <em>WNT</em> medulloblastomas still require to be described.</AbstractText><AbstractText>We performed a multicentric retrospective review of <em>12</em> patients treated between 1988 and 2018 for medulloblastoma with identified or highly suspected (personal of familial history) APC germline pathogenic variant. We report personal and familial history, APC gene pathogenic variant whenever available, clinical and histological characteristics of the medulloblastoma, treatments, and long-term outcome including second tumor and late sequelae.</AbstractText><AbstractText>Medulloblastomas associated with APC pathogenic variant are mainly classic (11/11 patients, 1 NA), non-metastatic (10/<em>12</em> patients) medulloblastomas, with nuclear immunoreactivity for ß-catenin (9/9 tested cases). 10/11 assessable patients are disease-free with a median follow-up of 10.7 years [1-28]. Secondary tumors included desmoid tumors in 7 patients (9 tumors), 1 thyroid carcinoma, 2 pilomatricomas, 1 osteoma, 1 vertebral hemangioma, and 1 malignant Triton in the radiation field which caused the only cancer-related death in our series.</AbstractText><AbstractText>Medulloblastomas associated with APC pathogenic variant have an overall favorable outcome, even for metastatic tumors. Yet, long-term survival is clouded by second tumor occurrence; treatment may play some role in some of these second malignancies. Our findings raise the question of applying de-escalation therapeutic protocol to treat patients with APC germline pathogenic variant given the excellent outcome, and reduced intensity of craniospinal irradiation may be further evaluated.</AbstractText>

<AbstractText>Spinal cord injury (SCI) is a neurological disease with high incidence and disability. In this paper, we made a foundational study in long non-coding RNA paternally expressed gene 10 (lncRNA PEG10) at PC-<em>12</em> cells.</AbstractText><AbstractText>We used CCK8, flow cytometry, migration and invasion assay to detect changes at the cellular level. PEG10, microRNA-34a-5p (miR-34a-5p) and TLX transfected by transfection assay and amplified by real-time quantitative PCR (qRT-PCR). TLX expression and proteins about apoptosis and pathways were investigated by Western blot. Additionally, the relationship between PEG10 and miR-34a-5p, miR-34a-5p and TLX were examined through luciferase assay.</AbstractText><p><div><b>RESULTS</b></div>H₂O₂-injury model was established. Knockdown PEG10 deteriorated H₂O₂-injury. miR-34a-5p was confirmed as a target of PEG10 and miR-34a-5p inhibitor remitted PEG10-induced H₂O₂-injury. Moreover, TLX was confirmed as a target miR-34a-5p and TLX remitted H₂O₂-injury. At last, TLX worked in H₂O₂-injury via Smad and <em>Wnt</em>/β-catenin pathways.</p><p><div><b>CONCLUSION</b></div>Knockdown PEG10 remitted H₂O₂-injury of PC-<em>12</em> cells by targeting miR-34a-5p/TLX, and the behavior may be involved in Smad and <em>Wnt</em>/β-catenin pathways.</p>

OBJECTIVE

Spinal cord injury (SCI) is associated with modulation of different microRNAs (miRs). This study aims to explore the role of miR-25 in PC-<em>12</em> cells to reveal the potential of miR-25 in SCI treatment.

METHODS

SCI model was established in C57BL/6 mice, then miR-expression in the injured spinal cords were detected by qRT-PCR. PC-<em>12</em> cells were exposed to H2O2 conditions to establish an in vitro model of SCI. PC-<em>12</em> cells were transfected with expressing vector or antisense oligonucleotides (ASO) of miR-25. The effects of miR-25 expression on H2O2-induced oxidative damage was evaluated by detection of cell viability, apoptosis, ROS activity, HIF-α and γH2A expression, and the level of inflammatory mediators. The expression of Nrf2 in cells was silenced by transfection with Nrf2 siRNA, and the effects of Nrf2 silence on miR-25-mediated PC-<em>12</em> cells were detected. Besides, the expression of main proteins in Wnt/β-catenin and PI3 K/AKT/ERK signaling were assessed.

RESULTS

miR-25 was low expressed in injured spinal cords. miR-25 protected PC-<em>12</em> cells against H2O2-induced oxidative damage, as evidenced by significant suppression in cell apoptosis, increase in cell viability, decrease in the level of ROS, HIF-α and γH2A, and decrease in inflammatory mediators (IL-1β, TNF-α, IL-6, and MCP-1). However, Nrf2 silence abolished the protective functions of miR-25 on H2O2-induced damage. Furthermore, we found that Wnt/β-catenin and PI3 K/AKT/ERK signaling were activated by miR-25.

CONCLUSIONS

miR-25 protects PC-<em>12</em> cells against H2O2-induced oxidative damage though regulation of Nrf2 and activation of Wnt/β-catenin and PI3 K/AKT/ERK signaling.

<AbstractText>Livestock production aims to provide meats of high and consistent eating quality. Insufficient intramuscular (IM) fat and excessive subcutaneous (SC) fat are paramount pork quality challenges. IM fat and SC fat, which are modulated by the adipogenesis of IM and SC adipocytes, play key roles in pork quality. Galectin-<em>12</em> (LGALS<em>12</em>) was proven to be an important regulator of fat deposition in porcine. However, the current knowledge of the transcriptome-wide role of LGALS<em>12</em> in adipocytes is still limited. This study was aimed to discover the different regulatory mechanisms of LGALS<em>12</em> in porcine IM and SC adipocyte.</AbstractText><AbstractText>The siRNA-mediated knockdown of the expression of LGALS<em>12</em> identified 1075 and 3016 differentially expressed genes (DEGs) in IM and SC adipocytes, respectively. Among these, 585 were up- and 490 were downregulated in the IM adipocytes, while 2186 were up- and 830 were downregulated in the SC adipocytes. Moreover, 418 DGEs were observed only in the IM adipocytes, 2359 DGEs only in the SC adipocytes, and 657 DGEs in both types of adipocytes. According to Gene Ontology (GO) analysis, DEGs in both IM and SC adipocytes were mainly enriched in categories related to lipids or fat cell differentiation. Pathway analysis of the DEGs revealed 88 changed signaling pathways in the IM adipocytes and 86 in the SC adipocytes. The signaling pathways present in only one type of adipocyte were identified from among the top 50 signaling pathways in each type of adipocyte. Four signaling pathways, encompassing PI3K-AKT, cardiac muscle contraction, fatty acid metabolism and Ras, were significantly enriched in the IM adipocytes. On the other hand, four different signaling pathways, encompassing TNF, <em>WNT</em>, cGMP-PKG and NF-kappa B, were greatly enriched in the SC ones. The pathway changes were confirmed by chemical inhibition assays.</AbstractText><AbstractText>Our data reveals that LGALS<em>12</em> knockdown alters the expression of numerous genes involved in key biological processes in the development of adipocytes. These observations provide a global view of the role of LGALS<em>12</em> in porcine IM and SC adipocytes; thus, improving our understanding of the regulatory mechanisms by which this gene acts in fat development.</AbstractText>

Extraskeletal osteosarcoma (ESOSA) is a rare soft tissue neoplasm representing <5% of osteosarcomas and <1% of all soft-tissue sarcomas. Herein, we investigate the clinicopathological and molecular features of ESOSA and explore potential parameters that may affect outcome. Thirty-two cases were retrieved and histomorphology was reviewed. Clinical history and follow-up were obtained through electronic record review. DNA from formalin-fixed paraffin-embedded (FFPE) tissue was extracted and processed from 27 cases. Genome-wide DNA copy number (CN) alterations and allelic imbalances were analyzed by single nucleotide polymorphism array using Affymetrix OncoScan FFPE Assay. Massive high-throughput deep parallel sequencing was performed using a customized panel targeting 410 cancer genes. Log rank, Fisher's exact test and Cox proportional hazards were used for statistical analysis. In this series of 32 patients (male n = <em>12</em>, female n = 20), the average age was 66 years (19-93) and median follow up was 24 months (range 6-<em>12</em>0 months). Frequent genomic alterations included CN losses in tumour suppressor genes including CDKN2A (70%), TP53 (56%) and RB1 (49%). Mutations affecting methylation/demethylation, chromatin remodeling and <em>WNT</em>/SHH pathways were identified in 40%, 27%, and 27%, respectively. PIK3CA and TERT promoter variant mutations were identified in 11% of the cases. Cases harbouring simultaneous TP53 and RB1 biallelic CN losses were associated with worse overall survival and local recurrence (p = 0.04, p = 0.02, respectively). CDKN2A losses and positive margins were also associated with worse overall survival (p = 0.002; p = 0.03, respectively). Our findings suggest that age above 60, positive margin status, simultaneous biallelic TP53 and RB1 losses and CDKN2A loss are associated with a worse outcome in ESOSA. Comparison between conventional paediatric osteosarcoma and ESOSA shows that, while both share genetic similarities, there are notable dissimilarities and mechanistic differences in the molecular pathways involved in ESOSA.

<AbstractText>Identification and validation of new functionally relevant and pharmacologically actionable targets for pancreatic ductal adenocarcinoma (PDAC) remains a great challenge. Premalignant acinar cell reprogramming (acinar-to-ductal metaplasia [ADM]) is a precursor of pancreatic intraepithelial neoplasia (PanIN) lesions that can progress to PDAC. This study investigated the role of proline-rich tyrosine kinase 2 (PYK2) in mutant Kras-induced and pancreatitis-associated ADM and PanIN formation, as well as in PDAC maintenance.</AbstractText><AbstractText>Genetically engineered mouse models of mutant Kras (glycine <em>12</em> to aspartic acid) and Pyk2 deletion were used for investigating the role of PYK2 in PDAC genesis in mice. In vitro ADM assays were conducted using primary pancreatic acinar cells isolated from mice. Immunohistochemistry, immunofluorescence, and a series of biochemical experiments were used to investigate upstream regulators/downstream targets of PYK2 in pancreatic carcinogenesis. PDAC cell line xenograft experiments were performed to study the role of PYK2 and its downstream target in PDAC maintenance.</AbstractText><p><div><b>RESULTS</b></div>PYK2 was increased substantially in ADM lesions induced by mutant Kras or inflammatory injury. Pyk2 deletion remarkably suppressed ADM and PanIN formation in a mutant Kras-driven and pancreatitis-associated PDAC model, whereas PYK2 knockdown substantially inhibited PDAC cell growth in vitro and in nude mice. This study uncovered a novel yes-associated protein 1/transcriptional co-activator with PDZ binding motif/signal transducer and activator of transcription 3/PYK2/β-catenin regulation axis in PDAC. Our results suggest that PYK2 contributes to PDAC genesis and maintenance by activating the <em>Wnt</em>/β-catenin pathway through directly phosphorylating β-catenin^Y654.</p><AbstractText>The current study uncovers PYK2 as a novel downstream effector of mutant KRAS signaling, a previously unrecognized mediator of pancreatitis-induced ADM and a novel intervention target for PDAC.</AbstractText>

Human epidermal growth factor 2 (HER2) overexpression leads to aggressive mammary tumour growth. Although the prognosis of HER2⁺ tumours in humans is greatly improved using biologicals, therapy resistance, which may be caused by increased phosphatidyl-3-kinase (PI3K), rous sarcoma proto-oncogene (cSRC) or wingless-type MMTV integration site family (<em>Wnt</em>) activity, is a major concern. A recent analysis of <em>12</em> canine mammary cell lines showed an association between HER2/3 overexpression and phosphatase and tensin homologue (PTEN) deletion with elevated <em>Wnt</em>-signalling. <em>Wnt</em>-activity appeared to be insensitive to phosphatidyl-3-kinase (PI3K) inhibitors but sensitive to Src-I1. We hypothesized that <em>Wnt</em> activation, was caused by HER2/3-activated cSRC activation. The role of HER2/3 on <em>Wnt</em> signalling was investigated by silencing HER2/3 expression using specific small interfering RNA (siRNAs). Next, the effect of an epidermal growth factor receptor (EGFR)/HER2 tyrosine kinase inhibitor on <em>Wnt</em> activity and migration was investigated and compared to other tyrosine kinase inhibitors (TKIs) of related signalling pathways. Finally, two TKIs, a cSRC and a PI3K inhibitor, were investigated in a zebrafish xenograft model. Silencing of HER1-3 did not inhibit the intrinsic high <em>Wnt</em> activity, whereas the HER kinase inhibitor afatinib showed enhanced <em>Wnt</em> activity. The strongest inhibition of <em>Wnt</em> activity and cell viability and migration was shown by cSRC inhibitors, which also showed strong inhibition of cell viability and metastasis in a zebrafish xenograft model. HER2/3 overexpression or HER2/3-induced cSRC activation is not the cause of enhanced <em>Wnt</em> activity. However, inhibition of cSRC resulted in a strong inhibition of <em>Wnt</em> activity and cell migration and metastasis. Further studies are needed to unravel the mechanism of cSRC activation and cSRC inhibition to restore sensitivity to HER-inhibitors in HER2/3-positive breast cancer.