Search

AIMP2 interacts with the DIX domain of DVL and modulate Wnt/β-catenin signaling in an Aimp2 gene dosage dependent manner

Wnt/β-catenin (CTNNB1) signaling is crucial for the proliferation and maintenance of intestinal stem cells (ISC), but excessive activation leads to ISC expansion and eventually colorectal cancer. Thus, negative regulators are required to maintain optimal levels of Wnt/β-catenin signaling. Aminoacyl-tRNA synthetase–interacting multifunctional proteins (AIMP) function in protein synthesis, but have also been implicated in signaling cascades affecting angiogenesis, immunity, and apoptosis. In this study, we investigated the relationship between AIMP2 and Wnt/β-catenin signaling in a murine model of intestinal homeostasis and tumorigenesis. Hemizygous deletion of Aimp2 resulted in enhanced Wnt/β-catenin signaling, increased proliferation of cryptic epithelial cells, and expansion of ISC compartments. In an ApcMin/+ background, Aimp2 hemizygosity increased adenoma formation. Mechanistically, AIMP2 disrupted the interaction between AXIN and Dishevelled-1 (DVL1) to inhibit Wnt/β-catenin signaling by competing with AXIN. Furthermore, AIMP2 inhibited intestinal organoid formation and growth by suppressing Wnt/β-catenin signaling in an Aimp2 gene dosage-dependent manner. Collectively, our results showed that AIMP2 acts as a haploinsufficient tumor suppressor that fine-tunes Wnt/β-catenin signaling in the intestine, illuminating the regulation of ISC abundance and activity. Cancer Res; 76(15); 4559–68. ©2016 AACR.

The role of AIMP2 in intestinal Wnt signaling is dependent on β-catenin destruction complex.

Analysis of polyposis and tumor initiation in Aimp2+/-:ApcMin/+ intestine

Supplemental Materials and Methods and Supplemental figure legends

Aimp2 gene dosage negatively correlates with organoid growth

Aimp2+/- mice exhibit increased villi length, crypt depth, and colonic epithelial cell proliferation

The purpose of this retrospective study was to investigate the association between ipsilateral recurrence of ductal carcinoma in situ (DCIS) and radiomics features from DCIS and contralateral normal breast on contrast enhanced breast MR imaging. A total of 163 patients with DCIS who underwent preoperative MR imaging between January 2010 and December 2014 were included (training cohort; n = 117, validation cohort; n = 46). Radiomics features were extracted from whole tumor volume of DCIS on early dynamic T1-subtraction images and from the contralateral normal breast on precontrast T1 and early dynamic T1-subtraction images. After feature selection, a Rad-score was established by LASSO Cox regression model. Performance of Rad-score was evaluated by the receiver operating characteristic (ROC) curve and Kaplan Meier curve with log rank test. The Rad-score was significantly associated with ipsilateral recurrence free survival (RFS). The low-risk group with a low Rad-score showed higher ipsilateral RFS than the high-risk group with a high Rad-score in both training and validation cohorts (p < 0.01). The Rad-score based on radiomics features from DCIS and contralateral normal breast on breast MR imaging showed the potential for prediction of ipsilateral RFS of DCIS.

BACKGROUND In the analysis of electronic health records, proper labeling of outcomes is mandatory. To obtain proper information from radiologic reports, several studies were conducted to classify radiologic reports using deep learning. However, the classification of pneumonia in bilingual radiologic reports has not been conducted previously. OBJECTIVE The aim of this research was to classify radiologic reports into pneumonia or no pneumonia using a deep learning method. METHODS A data set of radiology reports for chest computed tomography and chest x-rays of surgical patients from January 2008 to January 2018 in the Asan Medical Center in Korea was retrospectively analyzed. The classification performance of our long short-term memory (LSTM)–Attention model was compared with various deep learning and machine learning methods. The area under the receiver operating characteristic curve (AUROC), area under the precision-recall curve, sensitivity, specificity, accuracy, and F1 score for the models were compared. RESULTS A total of 5450 radiologic reports were included that contained at least one pneumonia-related word. In the test set (n=1090), our proposed model showed 91.01% (992/1090) accuracy (AUROCs for negative, positive, and obscure were 0.98, 0.97, and 0.90, respectively). The top 3 performances of the models were based on FastText or LSTM. The convolutional neural network–based model showed a lower accuracy 73.03% (796/1090) than the other 2 algorithms. The classification of negative results had an F1 score of 0.96, whereas the classification of positive and uncertain results showed a lower performance (positive F1 score 0.83; uncertain F1 score 0.62). In the extra-validation set, our model showed 80.0% (642/803) accuracy (AUROCs for negative, positive, and obscure were 0.92, 0.96, and 0.84, respectively). CONCLUSIONS Our method showed excellent performance in classifying pneumonia in bilingual radiologic reports. The method could enrich the research on pneumonia by obtaining exact outcomes from electronic health data.

In this study, we investigated the role of microRNA-99a (miR-99a) in hepatitis C virus (HCV) replication and lipogenesis in hepatocytes. Cell-culture-derived HCV (HCVcc) infection caused down-regulation of miR-99a in Huh-7 cells, and the relative levels of miR-99a were significantly lower in the sera of the HCV-infected patients than in those of healthy controls. Transfection of miR-99a-5p mimics resulted in a decrease in the intracellular and secreted HCV RNA levels. It also caused a decreased mammalian target of rapamycin (mTOR) protein level and phosphorylation of its downstream targets in HCV-replicating cells. Sterol regulatory element binding protein (SREBP)-1c expression and intracellular lipid accumulation decreased when either miR-99a-5p mimics or si-mTOR was transfected in oleic acid-treated Huh-7 cells. Overexpression of mTOR rescued HCV RNA replication and lipid droplet accumulation in miR-99a-5p mimics-transfected HCV replicon cells. Our data demonstrated that miR-99a ameliorates intracellular lipid accumulation by regulating mTOR/SREBP-1c and causes inefficient replication and packaging of intracellular HCV.

Background/Aims: Hepatitis C virus (HCV) replicates in the peripheral blood mononuclear cells (PBMCs), leading to the production of type I interferons (IFNs). It is well known that the gene expression profile of PBMC is similar to that of the liver. The present study explored the dynamic gene expression profile of PBMCs collected from HCV-infected patients undergoing direct-acting antiviral (DAA) therapy. Methods: A prospective cohort comprising 27 patients under DAA therapy was formed. Expression level of IFN-β and its downstream interferon-stimulated genes (ISGs) was measured in PBMCs before and after DAA treatment. Furthermore, immunoblotting was performed to identify the signaling molecules involved in the expression of ISGs. Results: The pretreatment expression level of interferon-induced protein 44 (IFI44) and C-X-C motif chemokine ligand 10 (CXCL10) correlated with the pretreatment expression level of IFN-β. After DAA treatment, a significant decrease in the expression levels of IFN-β, IFI44, and CXCL10 was observed in the PBMCs. Furthermore, the pretreatment expression level of IFN-β and ISGs correlated with the level of signal transducer and activator of transcription 1 (STAT1) phosphorylation, and DAA treatment abrogated STAT1 phosphorylation. Conclusions: Pretreatment activation of IFN-β response is rapidly normalized after DAA treatment. The present study suggests that the decreased type I IFN response by the clearance of HCV might contribute to DAA-induced alleviation of extrahepatic manifestation of chronic HCV infection. (Clin Mol Hepatol 2018;24:302-310)

Metallic fuel has been considered for sodium-cooled fast reactors because it can maximize the uranium resources. It generates rare earth elements as fission products, where it is reported by aggravating the fuel–cladding chemical interaction at the operating temperature. Rare earth elements form a multicomponent alloy (Ce–Nd–Pr–La–Sm–etc.) during reactor operation, where it shows a higher reaction thickness than a single element. Experiments have been carried out by simplifying multicomponent alloys for mono or binary systems because complex alloys have difficulty in the analysis. In previous experiments, xCe-yNd was fabricated with two elements, Ce and Nd, which have a major effect on the fuel–cladding chemical interaction, and the thickness of the reaction layer reached maximum when the rare earth elements ratio was 1:1. The objective of this study is to evaluate the effect and relationship of rare earth elements on such synergistic behavior. Single and binary rare earth model alloys were prepared by selecting five rare earth elements (Ce, Nd, Pr, La, and Sm). In the single system, Nd and Pr behaviors were close to diffusion, and Ce showed a eutectic reaction. In the binary system, Ce and Sm further increased the reaction layer, and La showed a non-synergy effect. Keywords: Annealing, Diffusion, Fuel–Cladding Chemical Interaction, Metals, Scanning Electron Microscopy

// Eun Byul Lee 1, * , Min Gyeong Cheon 1, * , Jun Cui 1, 2 , Yoo Jin Lee 3 , Eun Kyoung Seo 3 and Ho Hee Jang 1, 4 1 Department of Health Sciences and Technology, Graduate School of Medicine, Gachon University, Incheon, South Korea 2 Department of Ophthalmology, Affiliated Hospital of Yanbian University, Yanji, Jilin, People’s Republic of China 3 College of Pharmacy, Graduate School of Pharmaceutical Sciences, Ewha Womans University, Seoul, South Korea 4 Department of Biochemistry, College of Medicine, Lee Gil Ya Cancer and Diabetes Institute, Gachon University, Incheon, South Korea * These authors contributed equally to this work Correspondence to: Ho Hee Jang, email: hhjang@gachon.ac.kr Keywords: anticancer drug, natural compound, reactive oxygen species (ROS), apoptosis, autophagy Received: August 01, 2017 Accepted: August 28, 2017 Published: September 18, 2017 ABSTRACT 8-Hydroxy-2-methoxy-1,4-naphthoquinone (HMNQ), a natural compound isolated from the bark of Juglans sinensis Dode, displays cytotoxic activity against various human cancer cells. However, the molecular mechanism of the anticancer effect is unclear. In this study, we examined the cytotoxic mechanism of HMNQ at the molecular level in human cancer cells. Cells were treated with HMNQ in a dose- or time-dependent manner. HMNQ treatment inhibited cell viability, colony formation and cell migration, indicating that HMNQ induced cancer cell death. HMNQ-treated cells resulted in apoptotic cell death through PARP-1 cleavage, Bax upregulation and Bcl-2 downregulation. HMNQ was also observed to induce autophagy by upregulating Beclin-1 and LC3. Furthermore, HMNQ induced reactive oxygen species (ROS) production, which was attenuated by the ROS scavengers, NAC and GSH. Finally, HMNQ increased expression of JNK phosphorylation and the JNK inhibitor SP600125 rescued HMNQ-induced cell death, suggesting that the cytotoxicity of HMNQ is mediated by the JNK signaling pathway. Taken together, our findings show that HMNQ exhibits anticancer activity through induction of ROS-mediated apoptosis and autophagy in human cancer cells. These data suggest the potential value of HMNQ as a natural anticancer drug.

Background/Aims: The role of Elk-3 in the epithelial-mesenchymal transition (EMT) during liver fibrogenesis remains unclear. Here, we determined the expression of Elk-3 in in vitro and in vivo models and in human liver fibrotic tissues. We also investigated the molecular relationships among Elk- 3, early growth response-1 (Egr-1), and the mitogen activated protein kinases (MAPK) pathway during EMT in hepatocytes. Methods: We established an in vitro EMT model in which normal mouse hepatocyte cell lines were treated with transforming growth factor (TGF)-β1 and a CCl4-induced liver fibrosis model. Characteristics of EMT were determined by evaluating the expression levels of related markers. The expression of Elk-3 and its target Egr-1 were analyzed using Western blotting. Gene silencing of Elk-3 was performed using an siRNA knockdown system. Results: The expression levels of mesenchymal markers were increased during TGF-β1-induced EMT of hepatocytes. The expression levels of Elk-3 and Egr-1 were significantly (p

Virus-like particles (VLPs) possess great potential for organ-specific transport of therapeutic agents due to their central cavity surrounded by viral capsid proteins and similar tropism to their original viruses. The N-terminal truncated second open reading frame (Nt-ORF2) of the hepatotropic hepatitis E virus (HEV) forms VLPs via self-assembly. In the present study, we investigated whether HEV-LPs could deliver foreign genes specifically to the liver. HEV-LPs were obtained from Nt-ORF2 expression in Huh7 cells that were transduced with recombinant baculoviruses and purified by continuous density gradient centrifugation. The purified HEV-LPs efficiently penetrated liver-derived cell lines and the liver tissues. To evaluate HEV-LPs as gene delivery tools, we encapsulated foreign plasmids in HEV-LPs with disassembly/reassembly systems. Green fluorescence was detected at higher frequency in liver-derived Huh7 cells treated with HEV-LPs bearing GFP-encoding plasmids than in control cells. Additionally, HEV-LPs bearing Bax-encoding plasmids induced apoptotic signatures in Huh7 cells. In conclusion, HEV-LPs produced in mammalian cells can encapsulate foreign genes in their central cavity and specifically transport these genes to liver-derived cells, where they are expressed. The present study could contribute to advances in liver-targeted gene therapy.

Background In the analysis of electronic health records, proper labeling of outcomes is mandatory. To obtain proper information from radiologic reports, several studies were conducted to classify radiologic reports using deep learning. However, the classification of pneumonia in bilingual radiologic reports has not been conducted previously. Objective The aim of this research was to classify radiologic reports into pneumonia or no pneumonia using a deep learning method. Methods A data set of radiology reports for chest computed tomography and chest x-rays of surgical patients from January 2008 to January 2018 in the Asan Medical Center in Korea was retrospectively analyzed. The classification performance of our long short-term memory (LSTM)–Attention model was compared with various deep learning and machine learning methods. The area under the receiver operating characteristic curve (AUROC), area under the precision-recall curve, sensitivity, specificity, accuracy, and F1 score for the models were compared. Results A total of 5450 radiologic reports were included that contained at least one pneumonia-related word. In the test set (n=1090), our proposed model showed 91.01% (992/1090) accuracy (AUROCs for negative, positive, and obscure were 0.98, 0.97, and 0.90, respectively). The top 3 performances of the models were based on FastText or LSTM. The convolutional neural network–based model showed a lower accuracy 73.03% (796/1090) than the other 2 algorithms. The classification of negative results had an F1 score of 0.96, whereas the classification of positive and uncertain results showed a lower performance (positive F1 score 0.83; uncertain F1 score 0.62). In the extra-validation set, our model showed 80.0% (642/803) accuracy (AUROCs for negative, positive, and obscure were 0.92, 0.96, and 0.84, respectively). Conclusions Our method showed excellent performance in classifying pneumonia in bilingual radiologic reports. The method could enrich the research on pneumonia by obtaining exact outcomes from electronic health data.

Wnt/β-catenin (CTNNB1) signaling is crucial for the proliferation and maintenance of intestinal stem cells (ISC), but excessive activation leads to ISC expansion and eventually colorectal cancer. Thus, negative regulators are required to maintain optimal levels of Wnt/β-catenin signaling. Aminoacyl-tRNA synthetase–interacting multifunctional proteins (AIMP) function in protein synthesis, but have also been implicated in signaling cascades affecting angiogenesis, immunity, and apoptosis. In this study, we investigated the relationship between AIMP2 and Wnt/β-catenin signaling in a murine model of intestinal homeostasis and tumorigenesis. Hemizygous deletion of Aimp2 resulted in enhanced Wnt/β-catenin signaling, increased proliferation of cryptic epithelial cells, and expansion of ISC compartments. In an ApcMin/+ background, Aimp2 hemizygosity increased adenoma formation. Mechanistically, AIMP2 disrupted the interaction between AXIN and Dishevelled-1 (DVL1) to inhibit Wnt/β-catenin signaling by competing with AXIN. Furthermore, AIMP2 inhibited intestinal organoid formation and growth by suppressing Wnt/β-catenin signaling in an Aimp2 gene dosage-dependent manner. Collectively, our results showed that AIMP2 acts as a haploinsufficient tumor suppressor that fine-tunes Wnt/β-catenin signaling in the intestine, illuminating the regulation of ISC abundance and activity. Cancer Res; 76(15); 4559–68. ©2016 AACR.

The Y-box binding protein-1 (YB-1) is a transcription/translation regulatory protein, and the expression thereof is associated with cancer aggressiveness. In the present study, we explored the regulatory effects of YB-1 during the transforming growth factor-β1 (TGF-β1)-induced epithelial-to-mesenchymal transition (EMT) in lung adenocarcinoma cells. Downregulation of YB-1 increased E-cadherin promoter activity, and upregulation of YB-1 decreased promoter activity, suggesting that the YB-1 level may be correlated with the EMT. TGF-β1 induced YB-1 expression, and TGF-β1 translocated cytosolic YB-1 into the nucleus. YB-1 overexpression promoted TGF-β1-induced downregulation of epithelial markers, upregulation of mesenchymal markers, and cell migration. Moreover, YB-1 overexpression enhanced the expression of E-cadherin transcriptional repressors via TGF-β1-induced Akt activation. Our findings afford new insights into the role played by YB-1 in the TGF-β1 signaling pathway.

Gene-associated with retinoid-interferon-induced mortality 19 (GRIM-19) targets multiple signaling pathways involved in cell death and growth. However, the role of GRIM-19 in the pathogenesis of hepatitis virus infections remains unexplored. Here, we investigated the restrictive effects of GRIM-19 on the replication of hepatitis C virus (HCV). We found that GRIM-19 protein levels were reduced in HCV-infected Huh7 cells and Huh7 cells harboring HCV replicons. Moreover, ectopically expressed GRIM-19 caused a reduction in both intracellular viral RNA levels and secreted viruses in HCVcc-infected cell cultures. The restrictive effect on HCV replication was restored by treatment with siRNA against GRIM-19. Interestingly, GRIM-19 overexpression did not alter the level of phosphorylated STAT3 or its subcellular distribution. Strikingly, forced expression of GRIM-19 attenuated an increase in intracellular lipid droplets after oleic acid (OA) treatment or HCVcc infection. GRIM-19 overexpression abrogated fatty acid-induced upregulation of sterol regulatory element-binding transcription factor-1 (SREBP-1c), resulting in attenuated expression of its target genes such as fatty acid synthase (FAS) and acetyl CoA carboxylase (ACC). Treatment with OA or overexpression of SREBP-1c in GRIM-19-expressing, HCVcc-infected cells restored HCV replication. Our results suggest that GRIM-19 interferes with HCV replication by attenuating intracellular lipid accumulation and therefore is an anti-viral host factor that could be a promising target for HCV treatment.

Liver cancer stem cells (LCSCs) have attracted attention because they cause therapeutic resistance in hepatocellular carcinoma (HCC). Understanding the metabolism of LCSCs can be a key to developing therapeutic strategy, but metabolic characteristics have not yet been studied. Here, we systematically analyzed and compared the global metabolic phenotype between LCSCs and non-LCSCs using transcriptome and metabolome data. We also reconstructed genome-scale metabolic models (GEMs) for LCSC and non-LCSC to comparatively examine differences in their metabolism at genome-scale. We demonstrated that LCSCs exhibited an increased proliferation rate through enhancing glycolysis compared with non-LCSCs. We also confirmed that MYC, a central point of regulation in cancer metabolism, was significantly up-regulated in LCSCs compared with non-LCSCs. Moreover, LCSCs tend to have less active fatty acid oxidation. In this study, the metabolic characteristics of LCSCs were identified using integrative systems analysis, and these characteristics could be potential cures for the resistance of liver cancer cells to anticancer treatments.

Nonalcoholic fatty liver disease (NAFLD) is a hepatic manifestation of metabolic syndrome. Studies have demonstrated that anthocyanin-rich foods may improve hyperlipidemia and ameliorate hepatic steatosis. Here, effects of Aronia melanocarpa (AM), known to be rich of anthocyanins, on hepatic lipid metabolism and adipogenic genes were determined. AM was treated to C57BL/6N mice fed with high fat diet (HFD) or to FL83B cells treated with free fatty acid (FFA). Changes in levels of lipids, enzymes and hormones were observed, and expressions of adipogenic genes involved in hepatic lipid metabolism were detected by PCR, Western blotting and luciferase assay. In mice, AM significantly reduced the body and liver weight, lipid accumulation in the liver, and levels of biochemical markers such as fatty acid synthase, hepatic triglyceride and leptin. Serum transaminases, indicators for hepatocyte injury, were also suppressed, while superoxide dismutase activity and liver antioxidant capacity were significantly increased. In FL83B cells, AM significantly reduced FFA-induced lipid droplet accumulation. Protein synthesis of an adipogenic transcription factor, peroxisome proliferator-activated receptor gamma 2 (PPAR gamma 2) was inhibited in vivo. Furthermore, transcriptional activity of PPAR gamma 2 was down-regulated in vitro, and mRNA expression of PPAR gamma 2 and its downstream target genes, adipocyte protein 2 and lipoprotein lipase were down-regulated by AM both in vitro and in vivo. These results show beneficial effects of AM against hepatic lipid accumulation through the inhibition of PPAR gamma 2 expression along with improvements in body weight, liver functions, lipid profiles and antioxidant capacity suggesting the potential therapeutic efficacy of AM on NAFLD.

In general, excessive water uptake of highly sulfonated aromatic hydrocarbon membranes limits their use in the fabrication of polymer electrolyte membrane fuel cells (PEMFCs). In this study, sulfonated poly(ether ether ketone) (SPEEK) was successfully crosslinked using 4-vinylbenzyl chloride as a crosslinking agent via electron beam (EB) irradiation and subsequent Friedel-Craft reaction. The crosslinking process significantly reduced the water uptake and methanol permeability of the SPEEK membranes, and greatly improved their dimensional stability and mechanical strength, although with a slight reduction in elongation at break. The process reduced the proton conductivity of the SPEEK membranes, but the proton conductivities of the crosslinked SPEEK membranes remained comparable to that of Nafion 212. The study results demonstrated the potential of the proposed crosslinking procedure to overcome excessive water uptake-induced problems of highly sulfonated aromatic hydrocarbon membranes.

Phytochemical investigation of the bark of Juglans sinensis Dode (Juglandaceae) led to the isolation of two active compounds, 8-hydroxy-2-methoxy-1,4-naphthoquinone (1) and 5-hydroxy-2-methoxy-1,4-naphthoquinone (2), together with 15 known compounds 3-17. All compounds were isolated from this plant for the first time. The structures of 1 and 2 were elucidated by spectroscopic data analysis, including 1D and 2D NMR experiments. Compounds 1-17 were tested for their cytotoxicity against the A549 human lung cancer cell line; compounds 1 and 2 exhibited significant cytotoxicity and additionally had potent cytotoxicity against six human cancer cell lines, MCF7 (breast cancer), SNU423 (liver cancer), SH-SY5Y (neuroblastoma), HeLa (cervical cancer), HCT116 (colorectal cancer), and A549 (lung cancer). In particular, breast, colon, and lung cancer cells were more sensitive to the treatment using compound 1. In addition, compounds 1 and 2 showed strong cytotoxic activity towards human breast cancer cells MCF7, HS578T, and T47D, but not towards MCF10A normal-like breast cells. They also inhibited the colony formation of MCF7, A549, and HCT116 cells in a dose-dependent manner. Flow cytometry analysis revealed that the percentage of apoptotic cells significantly increased in MCF7 cells upon the treatment with compounds 1 and 2. The mechanism of cell death caused by compounds 1 and 2 may be attributed to the upregulation of Bax and downregulation of Bcl2. These findings suggest that compounds 1 and 2 may be regarded as potential therapeutic agents against cancer.

One of the most important factors in the production of cartilage is transforming growth factor beta1 (TGF-beta1). To obtain sustained release of TGF-beta1, a cell-mediated gene therapy technique was introduced. We infected chondrocytes with a retroviral vector carrying the TGF-beta1 gene. The single clone derivative showed sustained TGF-beta1 secretion. It also showed constitutive type II collagen expression. Whereas the TGF-beta1 protein itself is unable to induce formation of cartilage in vivo, human chondrocytes engineered to express a retroviral vector encoding TGF-beta1 showed cartilage formation in vivo when cells were injected into nude mice intradermally. These data suggest that cell-mediated gene therapy using TGF-beta1 as a transgene would be a promising treatment for osteoarthritis.

Doublecortin is 40 kDa microtubule-associated phosphoprotein required for neuronal migration and differentiation in various regions of the developing central nervous system. We have investigated the expression and cellular localization of doublecortin in the developing rat retina using immunocytochemistry and Western blot analysis. The expression of doublecortin was high from embryonic day 18 (E18) until E20 and was low during the postnatal period. The doublecortin immunoreactivity first appeared in a few radially orientated cells in the mantle zone of the primitive retina at E15. From E16 onward, the immunoreactivity appeared in two different regions: the inner part of the retina and middle of the neuroblastic layer. In the inner part, the somata of cells in the ganglion cell layer, in the distal row of the neuroblastic layer and profiles in the inner plexiform layer showed doublecortin immunoreactivity up to postnatal day 1 (P1). Afterwards, the doublecortin immunoreactivity persisted in the inner plexiform layer until P15, although the intensity decreased gradually with the maturation of the retina. In the middle of the neuroblastic layer, doublecortin immunoreactivity appeared in the radially orientated cells. These cells transformed into horizontal cells. The doublecortin immunoreactivity persisted in these cells up to P21. Given these results, doublecortin may play an important role in the migration and differentiation of specific neuronal populations in developmental stages of the rat retina.

Background and aimsDespite the discovery of hepatitis C virus (HCV) entry factor, the mechanism by which it is regulated by miRNAs remains unclear. Adipose tissue-derived human mesenchymal stem cells (AT-hMSCs) have been widely used for differentiated hepatocyte-like cells (DHCs). Here, we established an in vitro HCV infection model using DHCs from AT-hMSCs and identified miRNAs that modulate HCV infectivity.MethodsAT-hMSCs were differentiated into DHCs using the conditional media, and evaluated for hepatocyte characteristics using RT-PCR, immunocytochemistry, periodic acid-Schiff staining, and a urea synthesis assay. The expression of HCV candidate receptors was also verified using immunocytochemistry. The levels of candidate miRNAs targeting HCV receptors were then determined by relative quantitative RT-PCR (rqRT-PCR). Finally, DHCs were infected using HCVcc and serum from HCV-infected patients, and infectivity of the virus was measured by rqRT-PCR and transmission electron microscopy (TEM).ResultsThe expected changes in morphology, function and hepatic gene expression were observed during hepatic differentiation. Moreover, the expression of candidate HCV entry factors and miR-27a were altered during hepatic differentiation. The infection and replication of HCV occurred efficiently in DHCs treated with HCVcc or infected with serum from HCV-infected patients. In addition, HCV infectivity was suppressed in miR-27a-transfected DHCs, due to the inhibition of LDLR expression by miR-27a.ConclusionsOur results demonstrate that AT-hMSCs are a good source of DHCs, which are suitable for in vitro cultivation of HCV. Furthermore, these results suggest that miR-27a modulates HCV infectivity by regulating LDLR expression.

Epidemiological evidence suggests a lower incidence of prostate cancer in Asian countries, where soy products are more frequently consumed than in Western countries, indicating that isoflavones from soy have chemopreventive activities in prostate cells. Here, we tested the effects of the soy isoflavone genistein on antioxidant enzymes in DU145 prostate cancer cells. Genistein significantly decreased reactive oxygen species levels and induced the expression of the antioxidant enzymes manganese (Mn) superoxide dismutase (SOD) and catalase, which were associated with AMP-activated protein kinase (AMPK) and phosphatase and tensin homolog deleted from chromosome 10 (PTEN) pathways. The induced expression of catalase, MnSOD, and PTEN were attenuated by pretreatment with a pharmacological inhibitor for AMPK, indicating the effects of genistein primarily depend on AMPK. Furthermore, PTEN is essential for genistein activity, as shown by PTEN transfection in PTEN-deficient PC3 cells. Thus, genistein induces antioxidant enzymes through AMPK activation and increased PTEN expression.

Acanthosis nigricans (AN) is characterized by symmetric, velvety, gray-brown hypertrophied plaques most commonly on the axillae and neck. Confluent and reticulated papillomatosis (CRP) is manifested by papules in the mid-chest that coalesce in the midline, and are arranged in a reticulated pattern peripherally. Both diseases are known to be related to endocrinopathies such as diabetes mellitus (DM), an insulin resistant state and obesity with unknown mechanisms. Herein, we presented three cases of concomitant acanthosis nigricans with confluent and reticulated papillomatosis in obese patients.

Background Mucoceles have diverse clinical and histological features. Their pathogenesis remains to be elucidated. Objective To determine the structural and/or pathogenic differences between two clinically different types of mucoceles. Methods Seventeen oral mucoceles were examined clinically and immunohistologically. The mucoceles were divided into two groups by their clinical manifestations: papular group (PG) and nodular group (NG). Results Histologically, granulation tissue formed more frequently in the NG group, while CD4 and CD8 positive cells were more abundant in the PG group. There were no significant differences in the tumor necrosis factor-alpha or the matrix metalloproteinases (MMP)-2 and 9, between the two groups. Conclusion There were significant differences in the depth of the lesions, granulation tissue formation and infiltrating T lymphocytes between the PG and NP type of mucoceles. These findings suggest that the clinical manifestations may be influenced by the type of inflammatory response and extracellular matrix remodeling.