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Myocardial ischemia/reperfusion injury (MIRI) is a significant challenge in the management of myocardial ischemic disease. Extensive evidence suggests that the macrophage-mediated inflammatory response may play a vital role in MIRI. Mesenchymal stem cells and, in particular, exosomes derived from these cells, may be key mediators of myocardial injury and repair. However, whether exosomes protect the heart by regulating the polarization of macrophages and the exact mechanisms involved are poorly understood. The present study aimed to determine whether exosomes secreted by bone marrow mesenchymal stem cells (BMSC-Exo) harboring miR-25-3p can alter the phenotype of macrophages by affecting the JAK2/STAT3 signaling pathway, which reduces the inflammatory response and protects against MIRI. An in vivo MIRI model was established in rats by ligating the anterior descending region of the left coronary artery for 30 min followed by reperfusion for 120 min, and BMSC-Exo carrying miR-25-3p (BMSC-Exo-25-3p) were administered through tail vein injection. A hypoxia-reoxygenation model of H9C2 cells was established, and the cells were cocultured with BMSC-Exo-25-3p in vitro. The results of the present study demonstrated that BMSC-Exo or BMSC-Exo-25-3p could be taken up by cardiomyocytes in vivo and H9C2 cells in vitro. BMSC-Exo-25-3p demonstrated powerful cardioprotective effects by decreasing the cardiac infarct size, reducing the incidence of malignant arrhythmias and attenuating myocardial enzyme activity, as indicated by lactate dehydrogenase and creatine kinase levels. It induced M1-like macrophage polarization after myocardial ischemia/reperfusion (I/R), as evidenced by the increase in iNOS expression through immunofluorescence staining and upregulation of proinflammatory cytokines through RT-qPCR, such as interleukin-1β (IL-1β) and interleukin-6 (IL-6). As hypothesized, BMSC-Exo-25-3p inhibited M1-like macrophage polarization and proinflammatory cytokine expression while promoting M2-like macrophage polarization. Mechanistically, the JAK2/STAT3 signaling pathway was activated after I/R in vivo and in LPS-stimulated macrophages in vitro, and BMSC-Exo-25-3p pretreatment inhibited this activation. The results of the present study indicate that the attenuation of MIRI by BMSC-Exo-25-3p may be related to JAK2/STAT3 signaling pathway inactivation and subsequent inhibition of M1-like macrophage polarization. [ABSTRACT FROM AUTHOR]

Myocardial infarction (MI) is one of the leading causes of death globally. The aim of the present study was to find valuable microRNAs (miRNAs/miRs) and target mRNAs in order to contribute to our understanding of the pathology of MI. miRNA and mRNA data were downloaded for differential expression analysis. Then, a regulatory network between miRNAs and mRNAs was established, followed by function annotation of target mRNAs. Thirdly, prognosis and diagnostic analysis of differentially methylated target mRNAs were performed. Finally, an in vitro experiment was used to validate the expression of selected miRNAs and target mRNAs. A total of 19 differentially expressed miRNAs and 1,007 differentially expressed mRNAs were identified. Several regulatory interaction pairs between miRNA and mRNAs were identified, such as hsa-miR-142-2p-long-chain-fatty-acid-CoA ligase 1 (ACSL1), hsa-miR-15a-3p-nicotinamide phosphoribosyltransferase (NAMPT), hsa-miR-33b-5p-regulator of G-protein signaling 2 (RGS2), hsa-miR-17-3p-Jun dimerization protein 2 (JDP2), hsa-miR-24-1-5p-aquaporin-9 (AQP9) and hsa-miR-34a-5p-STAT1/AKT3. Of note, it was demonstrated that ACSL1, NAMPT, RGS2, JDP2, AQP9, STAT1 and AKT3 had diagnostic and prognostic values for patients with MI. In addition, STAT1 was involved in the 'chemokine signaling pathway' and 'Jak-STAT signaling pathway'. AKT3 was involved in both the 'MAPK signaling pathway' and 'T cell receptor signaling pathway'. Reverse transcription-quantitative PCR validation of hsa-miR-142-3p, hsa-miR-15a-3p, hsa-miR-33b-5p, ACSL1, NAMPT, RGS2 and JDP2 expression was consistent with the bioinformatics analysis. In conclusion, the identified miRNAs and mRNAs may be involved in the pathology of MI. [ABSTRACT FROM AUTHOR]

The purpose of the present study was to explore the potential molecular signaling pathway mediated by the statin rosuvastatin in cultured human coronary artery endothelial cells (HCAECs) induced by CoCl2. CoCl2 was used to induce the apoptosis of HCAECs. Myocardial infarction rats were established and received statin or PBS treatment. Reverse transcription-quantitative PCR, western blotting, ELISA, TUNEL assay and immunohistochemistry were used to analyze the role of statin treatment. The results showed that rosuvastatin treatment decreased apoptosis of HCAECs induced by CoCl2 by increasing anti-apoptosis Bcl-xl and Bcl-2 expression, and decreasing pro-apoptosis Bax, Bad, caspase-3 and caspase-9 expression. The myocardial ischemia rat model demonstrated that rosuvastatin treatment decreased the mitochondrial reactive oxygen species, inflammation, mitochondrial damage, lipid catabolism, heart failure and the myocardial infarction areas, but improved the cardiac function indicators, right and left ventricular ejection fraction and increased expression levels of Janus kinase (JAK) and signal transducer and activator of transcription (STAT)3 in myocardial tissue. In conclusion, the results of the current study revealed that the statin rosuvastatin presents cardioprotective effects by activation of the JAK2/STAT3 signaling pathway. [ABSTRACT FROM AUTHOR]

The present study aimed to identify potentially important biomarkers associated with polycystic ovary syndrome (PCOS) by integrating DNA methylation with transcriptome profiling. The transcription (E-MTAB-3768) and methylation (E-MTAB-3777) datasets were retrieved from ArrayExpress. Paired transcription and methylation profiling data of 10 cases of PCOS and 10 healthy controls were available for screening differentially expressed genes (DEGs) and differentially methylated genes (DMGs). Genes with a negative correlation between expression levels and methylation levels were retained by correlation analysis to construct a protein-protein interaction (PPI) network. Subsequently, functional and pathway enrichment analyses were performed to identify genes in the PPI network. Additionally, a disease-associated pathway network was also established. A total of 491 overlapping genes, and the expression levels of 237 genes, were negatively correlated with their methylation levels. Functional enrichment analysis revealed that genes in the PPI network were mainly involved with biological processes of cellular response to stress, negative regulation of the biosynthetic process, and regulation of cell proliferation. The constructed pathway network associated with PCOS led to the identification of four important genes (SPP1, F2R, IL12B and RBP4) and two important pathways (Jak-STAT signaling pathway and neuroactive ligand-receptor interaction). Taken, together, the results from the present study have revealed numerous important genes with abnormal DNA methylation levels and altered mRNA expression levels, along with their associated functions and pathways. These findings may contribute to an improved understanding of the possible pathophysiology of PCOS. [ABSTRACT FROM AUTHOR]

The phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway is involved in the regulation of multiple cellular physiological processes by activating downstream corresponding effector molecules, which serve an important role in the cell cycle, growth and proliferation. This is a common phenomenon; overactivation of the pathway is present in human malignancies and has been implicated in cancer progression, hence one of the important approaches to the treatment of tumors is rational drug design using molecular targets in the PI3K/AKT signaling pathway. In brief, the present review analyzed the effects of the PI3K/AKT signaling pathway on certain gynecological cancer types. [ABSTRACT FROM AUTHOR]

Numerous reports have indicated that microRNA-93-5p (miR-93-5p) is involved in the development and progression of human cancer, including non-small cell lung, gastric and breast cancer; however, the role of miR-93-5p in retinoblastoma (RB) remains unknown. In the present study, it was reported that miR-93-5p expression levels were significantly upregulated in RB tissues compared with in normal tissues by reverse transcription-quantitative polymerase chain reaction. Furthermore, it was demonstrated via cell counting kit-8 and Transwell assays that knockdown of miR-93-5p significantly suppressed the proliferation, migration and invasion of RB cells, but promoted cellular apoptosis. Regarding the underlying mechanism, the present study reported that phosphatase and tensin homolog (PTEN) was a direct target of miR-93-5p in RB cells. Overexpression of miR-93-5p significantly inhibited the expression of PTEN; opposing results were observed when PTEN expression was downregulated. Furthermore, the present study revealed that PTEN expression levels were downregulated and were inversely correlated with that of miR-93-5p in RB tissues. Additionally, the present study demonstrated that knockdown of PTEN in miR-93-5p-depleted RB cells significantly reversed the effects of miR-93-5p on cell proliferation, migration and invasion; miR-93-5p knockdown was suggested to promote PTEN expression, consequently inhibiting the activation of phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway. Collectively, the results of the present study demonstrated that miR-93-5p may serve a role as an oncogene by modulating the PTEN/PI3K/AKT signaling pathway in RB, indicating that miR-93-5p may be a potential therapeutic target for the treatment of RB. [ABSTRACT FROM AUTHOR]

Hepatitis B core antigen (HBcAg) possesses unusual immunologic features. However, the biological roles and mechanisms of HBcAg in dendritic cell proliferation and apoptosis remain to be elucidated. In the present study, DC2.4 cells were treated with different concentrations of HBcAg (10, 20 and 30 µg/ml). MTT assay and flow cytometry (Annexin V/propidium iodide analysis) were performed to investigate changes in cell proliferation and apoptosis. Western blot analysis was conducted to examine the changes in nuclear factor (NF)-κB and protein kinase C (PKC) signaling pathways. NF-κB inhibitor pyrrolidine dithiocarbamate (PDTC) and PKC inhibitor Chelerythrine were used to block these two signaling pathways. It was identified that HBcAg increased proliferation and decreased apoptosis in a dose-dependent manner. Western blotting results demonstrated that HBcAg upregulated p-PKC, p-IκB, p-P65, tumor necrosis factor-α and B-cell lymphoma 2 (Bcl-2) levels, and downregulated cleaved caspase 3, demonstrating that HBcAg activated the PKC and NF-κB signaling pathways. NF-κB inhibitor PDTC reduced the effects of HBcAg on DC2.4 proliferation (0.6 fold vs. 0.25 fold) and apoptosis (0.43 fold vs. 0.17 fold), and on Bcl-2 expression levels. PKC inhibitor Chelerythrine reduced the biological effects of HBcAg; it reduced proliferation (0.67 fold vs. 0.23 fold) and upregulated apoptosis (0.43 fold vs. 0.13 fold). Chelerythrine also blocked NF-κB activity and the HBcAg-induced Bcl-2 increase, suggesting the effect on Bcl-2 from HBcAg was dependent on the PKC/NF-κB signaling pathway. In conclusion, HBcAg promoted proliferation and inhibited apoptosis through the PKC/NF-κB/Bcl-2 signaling pathway in DC2.4 cells. [ABSTRACT FROM AUTHOR]

Acute kidney injury (AKI) is a clinically common and severe complication of ischemia-reperfusion (I/R), associated with high morbidity and mortality rates, and prolonged hospitalization. Rapamycin is a type of macrolide, primarily used for anti-rejection therapy following organ transplantation and the treatment of autoimmune diseases. Rapamycin has been identified to exert a protective effect against AKI induced by renal I/R as an autophagy inducer. However, whether rapamycin preconditioning may relieve AKI following cerebral I/R (CIR) remains to be fully elucidated. The purpose of the present study was to investigate the effects of CIR on the renal system of rats and the role of rapamycin in AKI following CIR. In the present study, a CIR model was established in Sprague-Dawley rats via a 90-min period of middle cerebral artery occlusion and 24 h reperfusion, and pretreatment with an intraperitoneal injection of rapamycin (dosage: 1 mg/kg; 0.5 h) prior to CIR. The levels of serum creatinine and blood urea nitrogen (BUN), and the expression of inflammation-, apoptosis- and autophagy-associated markers were subsequently measured. In addition to certain histopathological alterations to the kidney, it was identified that CIR significantly increased the levels of serum creatinine, BUN, tumor necrosis factor-α and interleukin-1β, and significantly induced apoptosis and autophagy. It was observed that rapamycin induced autophagy through the mammalian target of rapamycin complex 1/autophagy-related 13/unc-51 like autophagy activating kinase 1 signaling pathway, and that rapamycin pre-treatment significantly improved renal function and alleviated renal tissue inflammation and cell apoptosis in rats following CIR. In conclusion, the results suggested that rapamycin may alleviate AKI following CIR via the induction of autophagy. [ABSTRACT FROM AUTHOR]

Intervertebral disc degeneration (IDD) is a multifactorial disease with few efficacious clinical drugs, which has been demonstrated to be associated with nucleus pulposus (NP) cells apoptosis and degeneration of the extracellular matrix (ECM). Interleukin (IL)-1β, a common proinflammatory cytokine, is considered to be one of key regulators in IDD development. Andrographolide (AG), extracted from Andrographis paniculata, has been suggested to possess marked anti-inflammatory properties. However, the effects of AG on IDD has not been well explored. The present study aimed to investigate the effects and the mechanisms of AG on IDD in human NP cells. NP cells were treated with IL-1β in the absence or presence of AG to investigate the effects on cell viability, cellular apoptosis, production of ECM and matrix metalloproteinase (MMP)-3, MMP-9 and MMP-13, and a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)-4 and ADAMTS-5. It was identified that IL-1β-induced NP cellular apoptosis was significantly inhibited by AG treatment. Furthermore, AG mitigated the IL-1β-induced degeneration of the ECM, which was paralleled by a decrease in MMPs and ADAMTS levels. In addition, AG exhibited marked inhibitory properties against the activation of Toll-like receptors (TLRs), Myeloid differentiation factor 88 (MyD88) and the nuclear translocation of Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB). Taken together, these results demonstrated that AG treatment mitigated IL-1β-induced NP cells degeneration through the TLR4/MyD88/NF-κB signaling pathway, and suggested that AG may be a potential agent for IDD prevention and therapy. [ABSTRACT FROM AUTHOR]

The safety of anesthetics on the developing brain has caused concern. Ketamine, an N-methyl-D-aspartate receptor antagonist, is widely used as a general pediatric anesthetic. Recent studies suggested that ketamine alters the plasticity of dendritic spines in the developing brain and may be an important contributing factor to learning and cognitive impairment. However, the underlying molecular mechanism remains poorly understood. Therefore, the aim of the present study was to investigate the effect of ketamine on the plasticity of dendritic spines in cultured hippocampal neurons and the potential underlying mechanisms. After 5 days in vitro, rat hippocampal neurons were exposed to different concentrations (100, 300 and 500 µM) of ketamine for 6 h. Ketamine decreased the number and length of dendritic spines in a dose-dependent manner. Ketamine at a concentration of 300 µM caused an upregulation of transforming protein RhoA (RhoA) and Rho-associated kinase (ROCK) protein. These effects were inhibited by the ROCK inhibitor Y27632. These results suggested that ketamine induces loss and shortening of dendritic spines in hippocampal neurons via activation of the RhoA/ROCK signaling pathway. [ABSTRACT FROM AUTHOR]

Migration and invasion are the most important characteristics of human malignancies which limit cancer drug therapies in the clinic. Tongue squamous cell carcinoma (TSCC) is one of the rarest types of cancer, although it is characterized by a higher incidence, rapid growth and greater potential for metastasis compared with other oral neoplasms worldwide. Studies have demonstrated that the phenolic compound obovatol exhibits anti‑tumor effects. However, the potential mechanisms underlying obovatol‑mediated signaling pathways have not been completely elucidated in TSCC. The present study investigated the anti‑tumor effects and potential molecular mechanisms mediated by obovatol in TSCC cells and tissues. The results of the present study demonstrated that obovatol exerted cytotoxicity in SCC9 TSCC cells, and inhibited their migration and invasion. In addition, obovatol induced apoptosis in SCC9 TSCC cells by increasing caspase 9/3 and apoptotic protease enhancing factor 1 expression levels. Western blot analysis demonstrated that obovatol inhibited the expression of pro‑epidermal growth factor (EGF), Janus kinase (JAK), and signal transducer and activator of transcription (STAT) in SCC9 TSCC cells. A study of the molecular mechanisms demonstrated that depletion of EGF reversed the obovatol‑mediated inhibition of SCC9 TSCC cell growth and aggressiveness. Animal experiments indicated that obovatol significantly inhibited TSCC tumor growth and increased the number of apoptotic cells in tumor tissues. In conclusion, the results of the present study provided scientific evidence that obovatol inhibited TSCC cell growth and aggressiveness through the EGF‑mediated JAK‑STAT signaling pathway, suggesting that obovatol may be a potential anti‑TSCC agent. [ABSTRACT FROM AUTHOR]

Salidroside, an active ingredient extracted from the Rhodiola rosea plant, has potential anti-tumor effects. However, the effects of salidroside on gastric cancer cell proliferation and migration remain unclear. In the present study, the inhibitory effects of salidroside on gastric cancer cell proliferation, migration and invasion and the molecular mechanisms underlying these effects were investigated. The human gastric cancer cell line, BGC-823, was treated with different concentrations of salidroside (200, 400 and 600 µg/ml). Cell proliferation was determined with Cell Counting Kit-8 and colony formation assays, and the migration and invasion of cells was detected by a wound healing and Transwell assay, respectively. Western blotting was performed to detect the levels of N-cadherin, E-cadherin and heat shock protein (HSP)70. In addition, the phosphorylation of proto-oncogene tyrosine-protein kinase Src (Src), protein kinase B (Akt), mitogen activated protein kinase 1 (ERK), signal transducer and activator of transcription (STAT)3 and focal adhesion kinase 1 (FAK) was examined by western blotting. The levels of matrix metalloproteinase (MMP)-2 and MMP-9 were determined by enzyme-linked immunosorbent assay kits. Levels of reactive oxygen species (ROS) in cells were measured by a fluorescence plate reader with dichloro-dihydro-fluorescein diacetate. The results indicated that salidroside significantly suppressed cell proliferation and colony formation, inhibited cell migration and invasion, increased E-cadherin expression and decreased N-cadherin, MMP-2 and MMP-9 expression. Furthermore, salidroside suppressed ROS production and subsequently reduced the phosphorylation of Src, Akt, ERK and FAK. Salidroside also inhibited HSP70 expression, and HSP70 overexpression reversed the inhibitory effects of salidroside on BGC-823 cell proliferation, migration and invasion. In conclusion, the present study revealed that salidroside inhibited the proliferation, migration and invasion of BGC-823 cells by downregulating ROS-mediated Src-associated signaling pathway activation and HSP70 expression. [ABSTRACT FROM AUTHOR]

Chronic obstructive pulmonary disease (COPD) is a persistent airway inflammation influenced by cigarette smoke. Previous studies have reported that Hedgehog (Hh) signaling is aberrantly activated by cigarette smoke and dysregulated in COPD. The present study explored the role of the Hh signaling pathway on the expression levels of certain inflammatory mediators in cigarette‑induced airway inflammation. Herein, a total of three A549 cell populations were generated: The A0 group as control cells, the A1 group cells treated with nicotine at a concentration of 10 μM for 12, 24 and 48 h, and the A2 group cultured simultaneously with nicotine and cyclopamine for the same duration. The total concentrations of the inflammatory mediators interleukin‑6 (IL‑6), IL‑8 and tumor necrosis factor (TNF)‑α, and an anti‑inflammatory cytokine, IL‑10, were assessed in all of the cells by ELISA and western blotting. The protein levels of sonic hedgehog (Shh), glioma‑associated oncoprotein 1 (Gli1) and Smoothened (Smo) in nicotine‑induced Hh signaling were also detected. The results indicated that A549 had increased levels of IL‑6, IL‑8 and TNF‑α when cultured with nicotine when compared with the control cells. By contrast, the expression levels of these inflammatory mediators decreased with varying degrees when treated with cyclopamine that blocked the Hh signaling pathway. The IL‑10 expression levels exhibited the reverse. The expressions of the Shh, Gli1 and Smo proteins were higher in the A1 group when compared with the control and decreased with cyclpoamine treatment. In conclusion, the Hh signaling pathway may partly have an impact on cigarette‑induced airway inflammation via the regulation of inflammatory mediators. Thus, blocking Hh signaling and diminishing the airway inflammation reaction may serve as a potential therapy for COPD. [ABSTRACT FROM AUTHOR]

To investigate the role of microRNA (miR)‑149 in the cardiac differentiation of mouse bone marrow mesenchymal stem cells (MSCs) in vitro, MSCs were infected with a lentivirus overexpressing miR‑149 and the effect on cardiac differentiation was determined. The quantitative polymerase chain reaction results demonstrated that miR‑149 promoted the expression of cardiac‑specific markers in MSCs. Western blotting and a luciferase activity assay demonstrated that disabled homolog 2 (Dab2) was a direct target of miR‑149. Dab2 ectopic expression and Wnt/β‑catenin signaling pathway inhibition was able to reverse the increased expression of cardiac‑specific markers induced by miR‑149. In conclusion, miR‑149 was able to target Dab2 and promote the cardiac differentiation of mouse MSCs in vitro, which depended upon the Wnt/β‑catenin signaling pathway. [ABSTRACT FROM AUTHOR]

Hepatitis B virus X protein (HBx) has been termed a viral oncoprotein, and is involved in the initiation and progression of hepatocellular carcinoma (HCC). Cyclooxygenase‑2 (COX‑2) and β‑catenin have been attributed to the oncogenic activity of HBx in HBV‑associated HCC. The present study aimed to determine whether there is crosstalk between COX‑2 and the Wnt/β‑catenin signaling pathway during HL‑7702‑HBx cell proliferation, and to investigate the associated underlying molecular mechanism. In the present study, cell proliferation assay, colony formation assay and flow cytometric analysis were used to detect the proliferative ability of cells. Reverse transcription‑quantitative polymerase chain reaction and western blotting were performed to examine the mRNA and protein expression of COX‑2, β‑catenin, cyclin‑D1 and c‑myc. The results demonstrated that HL‑7702‑HBx exhibited increased cell proliferation, higher colony formation efficiency and a shortened G1 period of the cell cycle. In addition, the mRNA and protein expression levels of COX‑2 were increased, and this was associated with HL‑7702‑HBx cell growth. Furthermore, the expression of β‑catenin and its target genes, cyclin‑D1 and c‑myc proto‑oncogene protein, was upregulated by HBx via COX‑2. Finally, HBx promoted HL‑7702 cell proliferation through the Wnt/β‑catenin signaling pathway. In conclusion, the primary finding of the present study was that HBx may promote HL‑7702 cell proliferation via the COX‑2/Wnt/β‑catenin pathway. Thus, it may be helpful to further investigate the molecular mechanism of HBV‑associated hepatocellular carcinoma. [ABSTRACT FROM AUTHOR]

Diabetic retinopathy (DR), one of the most common complications of late‑phase diabetes, is associated with the ectopic apoptosis of microvascular cells. Gastrodin, a phenolic glucoside derived from Gastrodia elata Blume, has been reported to have antioxidant and anti‑inflammation activities. The aim of the present study was to investigate the effects of gastrodin on high glucose (HG)‑induced human retinal endothelial cell (HREC) injury and its underlying mechanism. The results demonstrated that HG induced cell apoptosis in HRECs, which was accompanied by increased levels of reactive oxygen species production. Gastrodin treatment significantly alleviated HG‑induced apoptosis and oxidative stress. Furthermore, HG stimulation decreased the levels of SIRT1, which was accompanied by an increase in Toll‑like receptor 4 (TLR4) expression and the levels of phosphorylated nuclear factor (NF)‑κBp65. However, the administration of gastrodin significantly inhibited the activation of the sirtuin 1 (SIRT1)/TLR4/NF‑κBp65 signaling pathway in HRECs exposed to HG. Collectively, the present study demonstrated that gastrodin may be effective against HG‑induced apoptosis and its action may be exerted through the regulation of the SIRT1/TLR4/NF‑κBp65 signaling pathway. [ABSTRACT FROM AUTHOR]

The present study aimed to investigate the mechanism by which the Janus kinase (JAK)2/signal transducer and activator of transcription (STAT)3 signaling pathway mediates cerebral ischemia and the efficacy of pharmaceutical intervention. The rat model of middle cerebral artery occlusion (MCAO) was established and confirmed via assessment of changes in the expression of phosphorylated (p)‑JAK2, p‑STAT3, high‑mobility group box 1 (HMGB1), and inflammatory factors using ELISA and western blot analysis. The effects of JAK2/STAT3 inhibitor and curcumin on the expression of p‑JAK2, p‑STAT3, HMGB1, and inflammatory factors after cerebral ischemia were observed with ELISA, western blotting and immunohistochemical staining. The concentrations of tumor necrosis factor (TNF)‑α and HMGB1 in brain tissue homogenate of MCAO group were significantly higher than in the sham group (P<0.01). The concentration of p‑JAK2/JAK2 and p‑STAT3/STAT3 in the brain tissue homogenate of MCAO group was significantly higher than in the sham group (P<0.05). The concentrations of TNF‑α, interleukin (IL)‑1β, IL‑6, and HMGB1 in the group treated with STAT3 inhibitor (MCAO + rapamycin), JAK2 inhibitor (MCAO + AG490), and MCAO + curcumin were significantly lower than in the MCAO group (P<0.01), as well as the relative content of p‑JAK2/JAK2 and p‑STAT3/STAT3 (P<0.05). Inhibition of the JAK2/STAT3 signaling pathway, such as curcumin can reduce the expression of HMGB1 in brain tissue after cerebral ischemia, which can significantly reduce the inflammatory response after cerebral ischemia. [ABSTRACT FROM AUTHOR]

The aim of the present study was to elucidate the effects of blocking the calcium signaling pathway of astrocytes (ASs) on oxygen and glucose deprivation (OGD)-induced AS injury. The association between the changes in the concentrations of AS-derived transmitter ATP and glutamic acid, and the changes in calcium signaling under the challenge of OGD were investigated. The cortical ASs of Sprague Dawley rats were cultured to establish the OGD models of ASs. The extracellular concentrations of ATP and glutamic acid in the normal group and the OGD group were detected, and the intracellular concentration of calcium ions (Ca2+) was detected. The effects of 2'-deoxy-N6-methyl adenosine 3', 5'-diphosphate diammonium salt (MRS2179), a P2Y1 receptor antagonist, on the release of calcium and glutamic acid of ASs under the condition of OGD were observed. The OGD challenge induced the release of glutamic acid and ATP by ASs in a time-dependent manner, whereas elevation in the concentration of glutamic acid lagged behind that of the ATP and Ca2+. The concentration of Ca2+ inside ASs peaked 16 h after OGD, following which the concentration of Ca2+ was decreased. The effects of elevated release of glutamic acid by ASs when challenged by OGD may be blocked by MRS2179, a P2Y1 receptor antagonist. Furthermore, MRS2179 may significantly mitigate OGD-induced AS injury and increase cell survival. The ASs of rats cultured in vitro expressed P2Y1 receptors, which may inhibit excessive elevation in the concentration of intracellular Ca2+. Avoidance of intracellular calcium overload and the excessive release of glutamic acid may be an important reason why MRS2179 mitigates OGD-induced AS injury. [ABSTRACT FROM AUTHOR]

Radiotherapy has an important role in the comprehensive treatment of breast cancer. However, the clinical outcome of adjuvant radiotherapy may be limited due to intrinsic radioresistance, it is necessary to explore efficient radiosensitization methods that improve the clinical outcome of patients undergoing radiotherapy. The present study aimed to investigate whether the novel mechanistic target of rapamycin (mTOR) inhibitor Torin2 enhances the radiosensitivity of MCF-7 breast cancer cells. A Cell Counting Kit-8 (CCK-8) assay was performed to measure the effect of Torin2 on cell proliferation, while clonogenic assays were employed to determine the effect of Torin2 in combination with radiation on the proliferation of MCF-7 cells. The effect of Torin2 and/or radiation on the cell cycle was analyzed using flow cytometry. Furthermore, the protein expression of components of the phosphatidylinositol 3-kinase/Akt/mTOR pathway, and the expression of proteins involved in DNA damage repair, was measured by western blot analysis. The results demonstrated that Torin2 exhibited a higher potency in MCF-7 cells, while MDA-MB-231 cells were less sensitive to Torin2. Compared with irradiation alone, pretreatment with 20 nM Torin2 followed by irradiation resulted in an increased level of γ-H2A histone family member X. Radiation induced the activation of the Akt/mTOR signaling pathway and upregulated the expression of phosphorylated (p)-Akt473 and p-eukaryotic translation initiation factor 4E binding protein 1 (4EBP1)37/46. Notably, pretreatment with Torin2 attenuated the radiation-induced activation of the Akt/mTOR signaling pathway. In addition, Torin2 partially blocked the repair of double-strand breaks induced by radiation by reducing the activation of ataxia telangiectasia-mutated, and sensitized MCF-7 cells to radiation. In conclusion, administration of Torin2 prior to irradiation enhanced the radiotherapeutic effect on breast cancer cells in vitro, and these results may provide a foundation for the rational use of combined therapy with irradiation and Torin2 for breast cancer in clinical practice. [ABSTRACT FROM AUTHOR]

Sepsis is the most important predisposing cause inducing acute respiratory distress syndrome (ARDS); however, the mechanism of sepsis leading to the development of ARDS remains to be elucidated. Suppression of the mitogen-activated protein kinase (MAPK) signal by blocking the phosphorylation of Jun N-terminal kinase (JNK) and p38 in lung tissues could alleviate acute lung injury induced by sepsis. MAPK signaling may have a crucial role in development of the sepsis-induced acute lung injury. The specific inhibitors of JNK and p38 MAPK, SP600125 and SB203580, were administrated by intragastric injection 4 h before induction of ARDS after cecal ligation and puncture (CLP). Rats were sacrificed at 1, 6 or 24 h after CLP challenge. The histological evaluation, lung water content, and biochemical analysis were performed. The results revealed that the JNK and p38 MAPK inhibitor improved lung permeability, attenuated system inflammation, further alleviated the lung injury induced by sepsis. In conclusion, JNK and p38 MAPK signaling are essential for the development of ARDS following sepsis. Further studies are needed to illuminate the detailed mechanisms of JNK and p38 MAPK signaling in sepsis-induced ARDS. [ABSTRACT FROM AUTHOR]

Rheumatoid arthritis (RA) is a chronic inflammatory articular disease that is characterized by synovial hyperplasia. A number of signaling pathways are associated with the development and induced symptoms of RA. Notably, patients with RA have increased protein citrullination and generation of auto-antibodies against citrullinated proteins. Genome wide association studies have revealed that peptidyl-arginine deiminase 4 (PADI4) is an enzyme implicated in citrullination in the RA synovium. Autoantibodies targeting citrullinated proteins are used as diagnostic markers in patients with RA. The functions associated with citrullinated proteins are thought to induce autoimmunity, however, the regulatory mechanisms of citrullination via PADI4 are unclear. The group has previously cloned an E3 ubiquitin ligase, synoviolin (SYVN1), from the RA synovium, demonstrating that SYVN1 serves critical roles in synovial hyperplasia. The data indicated that the endoplasmic reticulum (ER) associated degradation system, which involves SYVN1, may have important roles in the proliferation of synoviocytes. In addition, ubiquitination by SYVN1 is associated with fibrosis, inflammation and cytokine production via the regulation of ER stress signals and quality control of proteins. The present study investigated the crosstalk between the representative post-translational signaling processes, citrullination and ubiquitination. The results revealed that PADI4 interacted with SYVN1 directly and that overexpression of PADI4 suppressed the ubiquitination of proteins. Thus, a reduction in ER stress induced by PADI4 may abrogate the initiation of chronic RA by suppressing the proliferative signals of RA synoviocytes. [ABSTRACT FROM AUTHOR]

Increasing evidence indicates the important role of inflammation in the pathogenesis and progression of osteoarthritis (OA). Dual specificity phosphatase 1 (DUSP1), a negative regulator of the mitogen-activated protein kinase (MAPK) signaling pathway, has anti-inflammatory properties. In the present study, the expression of DUSP1 was investigated in human OA fibroblast-like synoviocytes (FLSs), human normal FLSs and OA FLSs pretreated with dexamethasone at the mRNA and protein levels. Then, the activation of MAPK pathway proteins and the expression of matrix metalloproteinase-13 (MMP-13) and cyclooxygenase-2 (COX-2) were measured by western blot analysis in the three groups of cells. Dexamethasone induced the expression of DUSP1 and inhibited the activation of the MAPK pathway and reduced the expression of MMP-13 and COX-2 in OA FLSs. However, the role of DUSP1 remained unclear. To clarify this, the effects of overexpression of DUSP1 in OA FLSs were determined using a DUSP1-overexpressing lentivirus. The results demonstrated that overexpression of DUSP1 in OA FLSs inhibited the activation of the MAPK pathway and expression of OA-associated mediators. The findings of the present study indicate that DUSP1 has a protective role in OA FLSs and may be a potential target in the treatment of OA. [ABSTRACT FROM AUTHOR]

The platelet-derived growth factor (PDFG) signaling pathway exerts persistent activation in response to a variety of stimuli and facilitates the progression of hepatic fibrosis. Since this pathway modulates a broad spectrum of cellular processes, including cell growth, differentiation, inflammation and carcinogenesis, it has emerged as a therapeutic target for hepatic fibrosis and liver-associated disorders. The present review exhibits the current knowledge of the role of the PDGF signaling pathway and its pathological profiles in hepatic fibrosis, and assesses the potential of inhibitors which have been investigated in the experimental hepatic fibrosis model, in addition to the clinical challenges associated with these inhibitors. [ABSTRACT FROM AUTHOR]

The role of inflammation in tumorigenesis and development is currently well established. Lymphotoxin β receptor (LTβR) activation induces canonical and noncanonical nuclear factor (NF)-κB signaling pathways, which are linked to inflammation-induced carcinogenesis. In the present study, 5,637 bladder cancer cells were cultured and the activation of LTβR was induced by functional ligand, lymphotoxin (LT) a1β2, and silencing with shRNA. Reverse transcription-quantitative polymerase chain reaction was utilized to detect the mRNA expression levels of NF-κB family members RelA and RelB, cytokines including LTα, LTβ, tumor necrosis factor (TNF)α, TNF superfamily member 14, interleukin (IL)-6 and IL-1β, and proliferation-related genes including CyclinD1 and Survivin. The expression of phospho-p65 was determined by western blotting. Activation of LTβR on bladder cancer 5,637 cells was demonstrated to upregulate the mRNA expression levels of the RELA proto-oncogene, RelA, by 2.5-fold compared with unstimulated cells, while no significant change was observed in the RELB proto-oncogene NF-κB member mRNA levels. Expression of pro-inflammatory cytokines tumor necrosis factor (TNF)α and interleukin (IL)-1β mRNA levels were significantly increased nearly 5-fold and 1.5-fold, respectively, following LTβR activation compared with unstimulated cells. The LTβR-induced upregulation of RelA, TNFα and IL-1β was decreased by ~33, 27, and 26% respectively when LTβR was silenced via short hairpin RNA. Activation of LTβR had no effect on 5,637 cell growth, despite CyclinD1 and Survivin mRNA levels increasing by ~2.7 and 1.3-fold, respectively, compared with unstimulated cells. In conclusion, activation of LTβR induced the expression of RelA mRNA levels. LTβR activation might be an important mediator in promoting an inflammatory microenvironment in bladder cancer, via the upregulation of TNFα and IL-1β mRNA levels. LTβR may be a potential therapeutic target for bladder cancer. [ABSTRACT FROM AUTHOR]

The differences in molecular mechanisms between osteochondroma and bizarre parosteal osteochondromatous proliferation (BPOP) remain to be fully elucidated. In the present study, the differentially expressed genes between BPOP and osteochondroma were obtained from the Gene Expression Omnibus online database, and the associations among these genes were analyzed using the Database for Annotation, Visualization, and Integrated Discovery (DAVID) online bioinformatics software. The results revealed several differentially expressed genes between human BPOP and osteochondroma. These differentially expressed genes were also enriched in different subgroups based on the analysis using DAVID online software, including 'transforming growth factor β receptor signaling pathway', 'BMP signaling pathway', 'Wnt receptor signaling pathway', 'response to chemical stimulus', 'regulation of inflammatory response', 'response to stress', 'glycosaminoglycan binding', 'polysaccharide binding', 'extracellular matrix structural constituent' and 'growth factors binding'. Taken together, these findings led to the conclusion that different gene regulatory mechanisms exist between BPOP and osteochondroma. Environmental stimulation and inflammation may contribute to BPOP or osteochondroma, and differences in extracellular matrix may contribute to differences in biological characteristics between BPOP and osteochondroma. [ABSTRACT FROM AUTHOR]

The aim of the present study was to determine the toxic targets of proteins from Croton tiglium L. and to investigate the potential mechanism of their toxicity. The toxic targets were determined by oral medication and intraperitoneal injection. The median lethal dose of oral medication in mice was calculated using Bliss software (2,752.8-3,407.5 mg/kg), and that of intraperitoneal injection was 195.8-272.69 mg/kg. The results of histopathological examination demonstrated that the kidney was primarily impaired by intraperitoneal injection, with slight degeneration of renal tubular epithelial cells. As to oral medication, the digestive tract was primarily injured, which manifested as congestion, bleeding, serious edema and other symptoms. Oral administration of the proteins caused gastrointestinal edema by increasing the intestinal permeability. Severe edema was associated with the inflammatory response, therefore the association between the toxicity of the proteins and inflammation was investigated. The proinflammatory effects of the crude proteins on the release of inflammatory mediator prostaglandin E2 (PGE2) were evaluated through intraperitoneal injection and the production of proinflammatory cytokines in RAW264.7 macrophages. Maximum PGE2 was released in the mice in vivo following intraperitoneal injection with 400 mg crude protein/kg body weight. Proinflammatory cytokines in macrophages, including tumor necrosis factor-a and interleukin-1ß, were produced in dose- and time-dependent manners in vitro. furthermore, the expressions of cell signaling molecules were detected by western blotting. The inflammatory response induced by crude protein in macrophages was associated with the mitogen-activated protein kinase (MAPK) signaling pathway mainly including p38-MAPK, extracellular signal-regulated kinase 1/2 and c-Jun N-terminal kinase 1/2/3 and the activated p38-MAPK signaling pathway. However, extracellular signal-regulated kinase 1/2 and c-Jun N-terminal kinases 1-3 exhibited no significant response. [ABSTRACT FROM AUTHOR]

Tubular epithelial-myofibroblast transdifferentiation (TEMT) is important in the development of chronic renal failure. The present study investigated whether hepatocyte growth factor (HGF) inhibits TEMT, and whether this function may be associated with the inhibition of angiotensin II (AngII) and the Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) signaling pathway. Human HK-2 kidney proximal tubular cells were divided into 4 groups and treated with AngII (1x10-6 M), HGF (8x10-3 M), AngII plus HGF or control conditions, followed by an assessment of apoptosis induction and the expression levels of α-smooth muscle actin (α-SMA), which is a marker of TEMT. as well as the activation level of JAK2, phosphorylated (p)-JAK2, STAT3 and p-STAT3 signaling pathways. In HK-2 cells, α-SMA mRNA and protein expression levels increased following treatment with AngII, however, decreased expression was observed following exposure to HGF. HGF counteracted the AngII-induced increase in the expression of α-SMA in HK-2 cells. Similar expression profiles were observed for the phosphorylated forms of JAK2 and STAT3, indicating the possible involvement of this signaling pathway. The results demonstrated that treatment of cells with AngII was associated with the induction of apoptosis when compared with the control. By contrast, treatment with HGF attenuated AngII-induced apoptosis. The results suggested that HGF may inhibit TEMT by inhibiting AngII through the JAK2/STAT3 signaling pathway in HK-2 cells and HGF may prevent apoptosis induced by AngII. The present study provides a basis for understanding the mechanisms involved in the inhibition of TEMT by HGF, which requires further investigation. [ABSTRACT FROM AUTHOR]

D-limonene has been demonstrated to have important immunomodulatory properties, including antitumor effects, and may alleviate asthma and allergies. In the present study, the anti-inflammatory effects of D-limonene were investigated in an ulcerative colitis (UC) rat model. Healthy male Sprague-Dawley rats were randomly divided into control, untreated UC, and treatment with 50 or 100 mg/kg D-limonene UC groups. In UC rats, disease activity and colonic mucosa damage were significantly reduced by the anti-inflammatory effects of D-limonene, via suppression of matrix metalloproteinase (MMP)-2 and -9 gene expression. In addition, treatment with D-limonene significantly increased antioxidant, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) protein expression levels in UC rats. A decrease in prostaglandin E2 (PGE2) production, transforming growth factor-β (TGF-β) gene expression and an increase phosphorylated-extracellular signal regulated kinase (ERK) 1/2 expression levelswere observed in UC rats treated with D-limonene. In conclusion, D-limonene reduced MMP-2 and -9 mRNA expression levels via regulation of the iNOS, COX-2, PGE2, TGF-β and ERK1/2 signaling pathways in a UC rat model, indicating its potential antioxidant and anti-inflammatory properties. [ABSTRACT FROM AUTHOR]

Inflammation in the retinal pigment epithelium is an important contributor to the pathogenesis of age-related macular degeneration. Wogonin is a flavonoid isolated from the root of Scutellaria baicalensis a nd has multiple pharmacological effects, including anti-inflammatory effects. The present study sought to determine if the pharmacological effects of wogonin were relevant to the treatment of AMD. ARPE-19 cells were pre-conditioned with different concentrations of wogonin (0-50 µM) prior to induction of inflammation with LPS (2 µg/ml). Transepithelial electrical resistance analysis demonstrated that 24 h treatment with 10 and 50 µM wogonin ameliorated LPS-induced changes. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and immunofluorescence analyses revealed that wogonin restrained LPS-induced tight junction proteins, claudin-1 and ZO-1. LPS-induced upregulation of inflammatory mediators in ARPE-19 cells, including IL-1β, IL-6, IL-8, cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS) and TNF-α was reduced after pre-treatment with wogonin. In addition, RT-qPCR and western blotting demonstrated that wogonin inhibited the expression of TLR4 in LPS-stimulated ARPE-19 cells. This is a novel mechanism indicating that pre-treatment with wogonin could attenuate the TLR4/NF-κB-mediated inflammatory response in LPS-stimulated ARPE-19 cells, and thus could be a potential therapy for the treatment of AMD. [ABSTRACT FROM AUTHOR]

Hypoxia is widely accepted as a fundamental biological phenomenon, which is strongly associated with tissue damage and cell viability under stress conditions. Insulin-like growth factor-1 (IGF-1) is known to protect tissues from multiple types of damage, and protect cells from apoptosis. Hypoxia is a regulatory factor of the IGF system, however the role of the IGF-1 receptor (IGF-1R) in hypoxia-induced apoptosis remains unclear. The present study investigated the potential mechanisms associated with IGF-1R-associated apoptosis under hypoxic conditions. Mouse embryonic fibroblasts exhibiting disruption or overexpression of IGF-1R (R- cells and R+ cells) were used to examine the level of apoptosis, autophagy, and production of reactive oxygen species (ROS). The autophagy inhibitor 3-methyladenine was used to assess the effect of autophagy on ROS production and apoptosis under hypoxic conditions. A potential downstream signaling pathway involving phosphatidylinositol 3-kinase (PI3K)/threonine protein kinase B (Akt)/mammalian target of rapamycin (mTOR) was identified by western blot analysis. The results demonstrated that hypoxia induced apoptosis, increased ROS production, and promoted autophagy in a time-dependent manner relative to that observed under normoxia. R+ cells exhibited a lower percentage of apoptotic cells, lower ROS production, and higher levels of autophagy when compared to that of R- cells. In addition, inhibition of autophagy led to increased ROS production and a higher percentage of apoptotic cells in the two cell types. Furthermore, IGF-1R is related with PI3K/Akt/mTOR signaling pathway and enhanced autophagy-associated protein expression, which was verified following treatment with the PI3K inhibitor LY294002. These results indicated that IGF-1R may increase cell viability under hypoxic conditions by promoting autophagy and scavenging ROS production, which is closed with PI3K/Akt/mTOR signaling pathway. [ABSTRACT FROM AUTHOR]

The present study aimed to determine whether there is an association between abnormalities in enteroendocrine cells in dextran sulfate sodium (DSS)-induced colitis and the clonogenic and/or proliferative activities of stem cells. A total of 48 male Wistar rats were divided into four groups. Animals in the control group were provided with normal drinking water, whereas DSS colitis was induced in the remaining three groups. The rats with DSS-induced colitis were randomized into the following three groups: i) DSS group, which received 0.5 ml 0.5% carboxymethyl cellulose (CMC; vehicle); ii) DSS-G group, which was treated with 3-[(dodecylthiocarbonyl)-methyl]-glutarimide at 20 mg/kg body weight in 0.5% CMC; and iii) DSS-Q group, which was treated with dehydroxymethylepoxyquinomicin at 15 mg/kg body weight in 0.5% CMC. Treatments were administered intraperitoneally twice daily for 5 days in all groups. Subsequently, tissue samples from the colon were stained with hematoxylin-eosin, or immunostained for chromogranin A (CgA), Musashi 1 (Msi1), Math-1, neurogenin 3 (Neurog3) and neurogenic differentiation D1 (NeuroD1). The densities of CgA, Msi1-, Math-1-, Neurog3- and NeuroD1-immunoreactive cells were determined. DTCM-G, and DHMEQ ameliorated the inflammation in DSS-induced colitis. The density of CgA-, Neurog3- and NeuroD1-immunoreactive cells was significantly higher in the DSS group compared with in the control group, and the density of CgA cells was correlated with the densities of Neurog3- and NeuroD1-immunoreactive cells. There were no significant differences in the densities of Msi1- and Math-1-immunoreactive cells among the four experimental groups. The elevated densities of enteroendocrine cells detected in DSS-induced colitis may be due to the increased differentiation of early enteroendocrine progenitors during secretory lineage. It is probable that the DSS-induced inflammatory processes trigger certain signaling pathways, which control differentiation of the stem-cell secretory lineage into mature enteroendocrine cells. [ABSTRACT FROM AUTHOR]

The traditional anti-diabetic drug, metformin, has been found to have anticancer effects. The Sonic hedgehog (Shh) signaling pathway is involved in the on cogenesis of gastric cancer. The aim of the present study was to investigate whether metformin has an effect on the Shh signaling pathway in gastric cancer cells. HGC-27 and MKN-45 human gastric cancer cells were treated with metformin at different concentrations and for different durations. Subsequently the mRNA and protein levels of Shh, Smoothened (SMO), and Glioma-associated oncogene (Gli)-1, Gli-2 and Gli-3 were examined using western blot and reverse transcription-quantitative polymerase chain reaction analyses. RNA interference was used to detect whether the effects of metformin treatment on the Shh signaling pathway were dependent on AMP-activated protein kinase (AMPK). The results revealed that the protein and mRNA levels of Shh and Gli-1 were decreased by metformin treatment in the two cell lines in a dose- and time-dependent manner. Metformin also significantly inhibited the gene and protein expression levels of SMO, Gli-2 and Gli-3. The small interfering RNA-induced depletion of AMPK reversed the suppressive effect of metformin on recombinant human Shh-induced expression of Gli-1 in HGC-27 gastric cancer cells. Therefore, metformin inhibited the Shh signaling pathway in the gastric cancer cell lines and the inhibitory effect of metformin on the Shh pathway was AMPK-dependent. [ABSTRACT FROM AUTHOR]

The aberrant expression of microRNA (miR)-214 contributes to the regulation of normal and cancer cell biology, and is associated with human malignancies, however, it can operate in a contradictory manner. The role of miR-214 in osteosarcoma remains to be fully elucidated. The aim of the present study was to investigate the effects of miR-214 on osteosarcoma progression and tumor cell proliferation, and examine the molecular mechanism underlying osteosarcoma. The level of miR-214 was determined using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis in osteosarcoma and matched paracancerous tissues, and in human osteosarcoma cancer cell lines. The roles of miR-214 in cell proliferation, survival and cell cycle were analyzed using miR-214 lentivirus (LV-miR-214)-infected osteosarcoma cells. In addition, the downstream target proteins in the Wnt/β-catenin signaling pathway were evaluated using western blot analysis in the LV-miR-214-infected cells. The LV-miR-214-infected MG63 cells were also treated with exogenous β-catenin for 24, 48 and 72 h, respectively, following which the expression of β-catenin was measured using western blot analysis and survival was determined using a 3-(4,5-cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The results of the RT-qPCR analysis showed that the expression level of miR-214 was significantly higher in the osteosarcoma tissues, compared with that in the matched paracancerous tissues, and the same was observed in the osteosarcoma cell lines. The MG63, Saos-2 and U2OS cells were infected with the hsa-mir-214 lentivirus for 48 h, and the levels of miR-214 were significantly upregulated in the human osteosarcoma cancer cells. The overexpression of miR-214 in the MG-63 and Saos-2 cells promoted cell growth, and treatment of the cells with specific antisense-microRNA oligonucleotides (AMOs) for miR-214 for indicated durations reversed the effects of miR-214. Additionally, the AMO-treated MG63 cells showed G0/G1 phase arrest, suggesting that miR-214 contributed to regulation of the cell cycle. In addition, the results of western blot analysis showed that, in the miR-214 lentivirus-infected cells, the levels of cyclin-D1, c-myc and lymphoid enhancer-binding factor-1 were significantly increased, compared with those in the control lentivirus-infected cancer cells. Of note, infection with the miR-214 lentivirus did not affect the levels of Wnt1, Wnt2, Wnt4, Axin or glycogen synthase kinase β in the U2OS cells, whereas the expression levels of β-catenin in the MG63 cells and Saos-2 cells were significantly increased. The addition of exogenous β-catenin effectively reversed the efficiency of miR-214-specific AMOs, which was detected using an MTT assay. These data suggested the critical role of miR-214 in human osteosarcoma via regulation of the Wnt/β-catenin signaling pathway and demonstrated that miR-214 is as an oncogene for human osteosarcoma. [ABSTRACT FROM AUTHOR]

In human leukaemia, resistance to chemotherapy leads to treatment ineffectiveness or failure. Previous studies have indicated that cancers with increased levels of aerobic glycolysis are insensitive to numerous forms of chemotherapy and respond poorly to radiotherapy. Whether glycolysis serves a key role in drug resistance of leukaemia cells remains unclear. The present study systematically investigated aerobic glycolytic alterations and regulation in K562/adriamycin (ADM) multidrug-resistant (MDR) and ADM-sensitive K562 leukaemia cells in normoxia, and the association between drug resistance and improper glycometabolism. The cell proliferating activity was assessed with an MTT colorimetric assay, glycolysis, including glucose consumption, lactate export and key-enzyme activity was determined by corresponding commercial testing kits. The expression levels of hexokinase-II (HK-II), lactate dehydrogenase A (LDHA), glucose transporter-4 (GLUT-4), AKT, p-AKT473/308, mammalian target of rapamycin (mTOR), p-mTOR, c-Myc and hypoxia-inducible factor-1α (HIF-1α) were analyzed by western blot or reverse transcription-quantitative polymerase chain reaction (RT-qPCR). K562/ADM cells exhibited increased glucose consumption and lactate accumulation, increased lactate dehydrogenase, hexokinase and pyruvate kinase activities, and reduced phosphofructokinase activity. In addition, K562/ADM cells expressed significantly more HK-II and GLUT-4. Notably, inhibition of glycolysis effectively killed sensitive and resistant leukaemia cells and potently restored the sensitivity of MDR cells to the anticancer agent ADM. The AKT serine/threonine kinase (AKT)/mechanistic target of rapamycin (mTOR) signalling pathway, a crucial regulator of glycometabolic homeostasis, mediated over-activation and upregulation of c-Myc expression levels in K562/ADM cells, which directly stimulated glucose consumption and enhanced glycolysis. In conclusion, the present study demonstrated that MDR leukaemia cells exhibit increased aerobic glycolytic activity and that this may be responsible for resistance to chemotherapeutics in leukaemia MDR cells via activation of the AKT-mTOR-c-Myc signalling pathway. Therefore, inhibition of aerobic glycolysis may be a potential therapeutic strategy to efficiently treat multidrug resistance in relapsed or refractory leukaemia and cancers. [ABSTRACT FROM AUTHOR]

Activated mast cells are involved in the pathogenesis of allergic rhinitis (AR). As a member of the matrix metalloproteinase (MMP) family, MMP9 has been previously demonstrated act in a pro-inflammatory manner. Mast cells regulate the activity of MMP9, and mast cells themselves have been reported to produce MMP9. However, to the best of our knowledge, the involvement of MMP9 in mast cell activation remains to be elucidated. The present study demonstrated an upregulation of MMP9 protein and mRNA expression levels in mast cells activated by phorbol ester and ionomycin. Phosphorylated ERK and AKT protein levels also markedly increased in activated mast cells, and inhibition of the ERK and AKT signaling pathways prevented the increase of MMP9 in activated mast cells. MMP9 was demonstrated to be involved in mast cell activation, since inhibition of MMP9 activity or expression inhibited mast cell activation. Furthermore, IL-4 treatment reduced MMP9 upregulation in activated mast cells, and interference with IL-4 signaling with an IL-4 neutralizing antibody promoted MMP9 upregulation in activated mast cells. These results revealed a novel MMP9-mediated mechanism underlying mast cell activation, thus providing novel ideas for AR therapy. [ABSTRACT FROM AUTHOR]

The present study investigated the effects of tauroursodeoxycholic acid (TUDCA) on the lipolytic action of tumor necrosis factor (TNF)-α in 3T3-L1 adipocytes. Following treatment with TNF-α, cell viability was determined by MTT assay to select the optimum concentration and duration of TNF-α treatment in 3T3-L1 adipocytes. Intracellular lipid droplet dispersion and glycerin content in culture media were determined to evaluate the effect of TUDCA on TNF-α-induced lipolysis in 3T3-L1 adipocytes. Western blotting was performed to detect protein expression levels of perilipin-A and protein markers of endoplasmic reticulum stress: Immunoglobulin-binding protein (BiP), inositol-requiring enzyme (IRE), c-Jun N-terminal kinase (JNK), phosphorylated (p)-IRE and p-JNK. Following treatment with 50 ng/ml TNF-α for 24 h, glycerin content increased significantly and lipid droplets were dispersed. Glycerin content was reduced significantly and dispersal of lipid droplets reduced following pretreatment of 3T3-L1 adipocytes with 1 mmol/l TUDCA. TNF-α additionally activated the expression of BiP, p-IRE and p-JNK in a time-dependent manner; following pretreatment of 3T3-L1 adipocytes with 1 mmol/l TUDCA, the expression levels of these three proteins decreased. Therefore, TUDCA may inhibit TNF-α-induced lipolysis in 3T3-L1 adipocytes and reduce production of free fatty acids. Its underlying molecular mechanisms are potentially associated with the inhibition of activation of the IRE-JNK signaling pathway, which influences perilipin-A expression levels. [ABSTRACT FROM AUTHOR]

A subset of basal cell carcinomas (BCCs) are directly derived from hair follicles (HFs). In some respects, HFs can be defined as 'ordered' skin appendage growths, while BCCs can be regarded as 'disordered' skin appendage growths. The aim of the present study was to examine HFs and BCCs to define the expression of common and unique signaling pathways in each skin appendage. Human nodular BCCs, along with HFs and non-follicular skin epithelium from normal individuals, were examined using microarrays, qPCR, and immunohistochemistry. Subsequently, BCC cells and root sheath keratinocyte cells from HFs were cultured and treated with Notch signaling peptide Jagged1 (JAG1). Gene expression, protein levels, and cell apoptosis susceptibility were assessed using qPCR, immunoblotting, and flow cytometry, respectively. Specific molecular mechanisms were found to be involved in the process of cell self-renewal in the HFs and BCCs, including Notch and Hedgehog signaling pathways. However, several key Notch signaling factors showed significant differential expression in BCCs compared with HFs. Stimulating Notch signaling with JAG1 induced apoptosis of BCC cells by increasing Fas ligand expression and downstream caspase-8 activation. The present study showed that Notch signaling pathway activity is suppressed in BCCs, and is highly expressed in HFs. Elements of the Notch pathway could, therefore, represent targets for the treatment of BCCs and potentially in hair follicle engineering. [ABSTRACT FROM AUTHOR]

Idiopathic pulmonary fibrosis (IPF) is the most common interstitial lung disease. However, the pathogenesis remains to be fully elucidated. Melatonin is secreted by the pineal gland, it has a strong antioxidant effect, and exerts an anti-fibrosis effect. Whether melatonin attenuates pulmonary fibrosis by inhibiting epithelial-mesenchymal transition (EMT) requires further research. The present study aimed to investigate whether melatonin prevents transforming growth factor-β1 (TGF-β1)-induced EMT and underlying signaling pathways using reverse transcription-quantitative polymerase chain reaction, western blot analysis and immunofluorescence. The results demonstrated that melatonin inhibits EMT in A549 cells, and the Wnt/β-catenin and Smad2/3 signaling pathways are involved in the EMT of the A549 cell line as they were suppressed by melatonin. The present study indicates that melatonin inhibited TGFβ1-induced epithelial-mesenchymal transition in the A549 cell line and may potentially be useful in the treatment of IPF. [ABSTRACT FROM AUTHOR]

Stroke is a life-threatening disease that results in significant disability in the human population. Despite the advances in current stroke therapies, a host of patients do not benefit from the conventional treatments. Thus, more effective therapies are required. It has been previously reported that leucine-rich-α2-glycoprotein 1 (LRG1) is crucial during the formation of new blood vessels in retinal diseases. However, the function of LRG1 in the brain during the neovessel growth process following ischemic stroke has not been fully elucidated and the mechanism underlying its effect on angiogenesis remains u nclear. T he purpose of t he current study was to demonstrate whether LRG1 may promote angiogenesis through the transforming growth factor (TGF)-β1 signaling pathway in ischemic rat brain following middle cerebral artery occlusion (MCAO). In the present study, the spatial and temporal expression of LRG1, TGF-β1, vascular endothelial growth factor (VEGF) and angiopoietin-2 (Ang-2) were detected in ischemic rat brain following MCAO using reverse transcription-quantitative polymerase chain reaction (RT-qPCR), western blot analysis and immunohistochemistry. CD34 immunohistochemistry staining was used as an indicator of microvessel density (MVD). The RT-qPCR and western blotting results revealed that the levels of LRG1 and TGF-β1 mRNA and protein expression were significantly increased as early as 6 and 12 h after MCAO (P<0.05), respectively, peaked at 3 days and persisted at significantly higher level until 14 days, in comparison with the control group. Additionally, VEGF and Ang-2 were also increased following MCAO. Furthermore, the immunohistochemistry results suggested that the MVD was increased following MCAO. In addition, the results also revealed that the percentage of LRG1-positive cells was positively correlated with the percentage of TGF-β1-positive cells, and the percentage of LRG1-positive and TGF-β1-positive cells had a positively correlation with the MVD. Taken together, the present study indicated that LRG1 may promote angiogenesis through upregulating the TGF-β1 signaling pathway in ischemic rat brain following MCAO. This may provide a potential therapeutic target for the treatment of ischemic stroke. [ABSTRACT FROM AUTHOR]

Ginkgolide B (GB) is one of the ginkgolides isolated from the leaves of the Ginkgo biloba tree. Our previous study indicated that GB promotes the proliferation, migration and adhesion of endothelial progenitor cells, and the induction of angiogenesis through vascular endothelial factor (VEGF). In the present study, the effects of GB on the differentiation of MC3T3-E1 cells and the signaling pathway involved were investigated in vitro. The MC3T3-E1 cell viability activities were assessed using an MTS assay. Measurements of alkaline phosphatase activity and Alizarin Red staining were used to identify osteoblastic differentiation of the MC3T3-E1 cells. The mRNA and secretion levels of VEGF were detected using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis and enzyme-linked immunosorbent assays, respectively. The protein expression levels of phosphorylation-associated markers were detected using western blot analysis and associated gene expression was determined using RT-qPCR analysis. It was found that GB significantly promoted alkaline phosphatase activity and osteoblastic mineralization in the MC3T3-E1 cells. In addition, the mRNA expression and secretion levels of VEGF in the MC3T3-E1 cells were significantly increased in MC3T3-E1 cells treated with GB. SB203580, a specific inhibitor of p38 mitogen-activated protein (MAP) kinase, markedly suppressed the GB-induced p38 kinase phosphorylation and GB-induced synthesis of VEGF. PD98059, an inhibitor of the upstream kinase, which activates p44/p42 MAP kinase, had minimal effect on the GB-induced phosphorylation of p44/p42 MAP kinase or the GB-induced synthesis of VEGF. Taken together, these results indicated that GB promoted osteoblastic differentiation of the MC3T3-E1 cells through VEGF, and that the p38, but not the p44/p42 MAP kinase signaling pathway, was involved in the GB-induced synthesis of VEGF. [ABSTRACT FROM AUTHOR]

Schiff base copper (II) complexes are known for their anticancer, antifungal, antiviral and anti-inflammatory activities. The aim of the current study was to investigate biological effects of Schiff base Cu (II) complexes (0.001-100 μmol/l)-[Cu2(sal-D, L-glu)2(isoquin oline)2]·2C2H5OH (1), [Cu(sal-5-met-L-glu)(H2O)].H2O (2), [Cu(ethanol)2(imidazole)4][Cu2(sal-D, L-glu)2(imidazole)2] (3), [Cu(sal-D,L-glu)(2-methylimidazole)] (4) on the human colon carcinoma cells HT-29, the mouse noncancerous cell line NIH-3T3 and the human noncancerous fibroblast cell line VH10. The results suggested that Cu (II) complexes exhibit cytotoxic effects against the HT-29 cell line, while complexes 3 and 4 were the most effective. Subsequent to 72 h of incubation, apoptosis was observed in the HT-29 cells induced by Cu (II) complexes 1 (0.1, 1, 10 and 50 μmol/l), 2 (1, 10, 50 and 100 μmol/l), 3 (0.01, 1, 10 and 50 μmol/l) and 4 (0.01, 0.1, 1 and 10 μmol/l). The apoptotic pathways activated by the Cu (II) complexes were identified. The results indicated that complexes 2, 3 and 4 were able to induce the mitochondria-dependent pathway of apoptosis in HT-29 cells, while complex 1 was obsered to activate the extrinsic pathway of apoptosis. The levels of the anti-apoptotic protein Bcl-2 were reduced and those of the pro-apoptotic protein Bax increased following treatment with complexes 2, 3 and 4. Complex 1 had no effect on Bax protein expression. Complexes 2 and 3 induced elevation of cytochrome c (cyt c), while complex 4 induced a time-dependent elevation of cyt c levels. No cyt c was detected in HT-29 cells exposed to complex 1, suggesting that Cu (II) complexes activated the extrinsic pathway of apoptosis. The results from the current study in addition to previous studies suggest that Schiff base Cu (II) complexes have potential as novel anticancer drugs. [ABSTRACT FROM AUTHOR]

Pulmonary arterial hypertension (PAH) is characterized by endothelial dysfunction and structural remodeling of the pulmonary vasculature, mediated initially by reduced oxygen availability in the lungs. Hypoxia inducible factor (HIF), consisting of the functional subunit, HIF-1α, and the constitutively expressed HIF-1β, is involved in the pathological processes associated with hypoxia. In the current study, the sequences of cDNAs and amino acids of HIF were characterized and analyzed using online bioinformatics tools. To further evaluate whether HIF accounts for the occurrence of PAH, the present study determine the expression and phosphorylation levels of HIF and its associated pathways, including extracellular signal-regulated kinase (Erk)1/2 and phosphoinositide 3-kinase (PI3K)/Akt, in the lungs of patients with PAH by reverse transcription-quantitative polymerase chain reaction and western blotting. The mRNA expression levels of PI3K, Erk2, and HIF-1α in the patients with PAH were significantly higher, compared with those in the control group, by 3.6-fold (P<0.01), 4.06-fold and 2.64-fold (P<0.05), respectively. No significant differences were found in the mRNA and protein levels of Akt between the two groups (P>0.05). The protein levels of phosphorylated (p-) Akt, Erk1/2, p-Erk1/2, HIF-1α and HIF-1β were significantly increased by 5.89-, 0.5-, 0.59-, 1.46- and 0.92-fold, respectively, in the patients with PAH, compared with those in the controls group (P<0.01 for p-Akt, Erk1/2; P<0.05 for p-Erk1/2, HIF-1α and HIF-1β). These findings suggested that the mitogen-activated protein kinase and PI3K/Akt signaling pathways, and HIF-1 may perform a specific function in the pathogenesis of PAH. [ABSTRACT FROM AUTHOR]

Metastasis is a major cause of lung cancer-associated mortality. The current study aimed to investigate the effects and mechanisms of TAR (human immunodeficiency virus 1) RNA binding protein 2 (TARBP2) in the invasion and migration of non-small cell lung cancer in vitro. The highly metastatic cell clone H1299/M02 was obtained by TARBP2 overexpression. Expression of TARBP2 in H1299/M02 was also downregulated to different levels via small hairpin RNAs (shRNAs). Subsequent to TARBP2 silencing, the proliferation of H1299/M02 cells was predominantly unaffected, while invasion and migration were significantly inhibited. A positive correlation was observed between invasion and migration and the level of TARBP2 silencing in vitro. Western blotting and reverse transcription-quantitative polymerase chain reaction indicated that the protein expression levels of amyloid β (A4) precursor protein (APP) and zinc finger protein 395 (ZNF395) were upregulated, while expression levels of pro-metastatic proteins including interleukin (IL)-1β, IL-8, cyclooxygenase (COX)-2, matrix metalloproteinase 2 (MMP2) and MMP9 were downregulated. Phosphorylation of c-Jun N-terminal kinase (JNK), signal transducer and activator of transcription 3 (STAT3) and protein kinase B (AKT) were also inhibited. Overexpression of TARBP2 was suggested to be involved in the metastasis of H1299/M02 cells. Silencing of TARBP2 was able to upregulate levels of APP and ZNF395, in addition to inhibiting metastasis-promoting cytokines, the JNK/STAT3/AKT pathway and COX-2 to attenuate the invasion and migration of cancer cells. [ABSTRACT FROM AUTHOR]

Prostate cancer is the fifth most common cause of cancer-associated mortality for males worldwide. Although dysregulation of the β-catenin/T-cell factor (TCF) pathway has been previously reported in prostate cancer, the mechanisms underlying this process remain unknown. Frequently rearranged in advanced T-cell lymphomas-1 (FRAT1) functions as a positive regulator of the β-catenin/TCF signaling pathway. However, to the best of our knowledge, the molecular association between FRAT1 and the β-catenin/TCF pathway in prostate cancer has not been investigated. In the present study, FRAT1 expression was analyzed in normal prostate tissues and prostate adenocarcinoma samples using publicly available databases, a commercial tissue microarray and immunohistochemistry techniques. In addition, FRAT1 expression levels were altered by overexpression or RNA interference-mediated depletion in prostate cancer cells. The effects of FRAT1 expression on tumor growth were determined using cell growth curves in vitro and xenografts in nude mice in vivo. The effects of FRAT1 on β-catenin/TCF activity were measured using the TOPFLASH reporter assay. FRAT1 was expressed exclusively in the nuclei of normal prostate basal cells, and nuclear FRAT1 was detected in 68% (40/59) of prostate adenocarcinoma samples. In addition, FRAT1 activated the TCF luciferase reporter gene promoter in prostate cancer cells, and was observed to promote the growth of prostate cancer cells in vitro. Furthermore, FRAT1 expression was sufficient to transform NIH3T3 mouse embryonic fibroblast cells and lead to tumor formation in vivo. These results suggest that FRAT1 demonstrates oncogenic properties in prostate cancer, potentially by suppressing the inhibitory effect of nuclear glycogen synthase 3β against β-catenin/TCF activity, thus activating the Wnt/β-catenin signaling pathway and promoting cell growth. [ABSTRACT FROM AUTHOR]

Livin, a member of the human inhibitor of apoptosis protein (IAP) family, is expressed at high levels in various human cancer tissues and may have prognostic significance. The aim of the present study was to evaluate the effect of Livin on tumor cell behavior and oncogenic signaling pathways in human hypopharyngeal squamous cell carcinoma (HSCC). Reverse transcription-quantitative polymerase chain reaction and western blot analyses were used to determine the mRNA and protein expression levels, respectively. A cell proliferation assay and cell cycle analysis were used to assess the functional effects of small interfering RNA-mediated Livin knockdown. Livin was overexpressed in fresh HSCC tissues, compared with the adjacent normal mucosa. Livin knockdown led to significantly reduced cell proliferation and cell cycle arrest in the G1 phase of the human HSCC cells. The expression levels of c-myc, cyclin D1, cyclin D3, cyclin-dependent kinase (CDK)4 and CDK6 were decreased. The phosphorylation levels of extracellular signal-regulated kinase 1/2, p38, c-Jun N-terminal kinase and Akt were also decreased by Livin knockdown in the HSCC cells. Taken together, the results of the present study suggested that Livin may enhance tumorigenesis by modulating the mitogen-activated/Akt signaling pathways in human HSCC. [ABSTRACT FROM AUTHOR]

Vascular smooth muscle cell (VSMC) proliferation and migration are critical in the progression of atherosclerosis and can be induced by platelet-derived growth factor (PDGF). Several studies have demonstrated that scavenger receptor class A, member 5 (SCARA5) is important in cancer cell migration and invasion. However, the role of SCARA5 in VSMCs remains to be elucidated in the development of atherosclerosis. Therefore, the role of SCARA5 was investigated in PDGF-BB-stimulated VSMC proliferation and migration. In the present study, it was shown that SCARA5 expression was enhanced by PDGF-BB in human aortic smooth muscle cells (HASMCs). Knockdown of SCARA5 by small interfering (si)RNA significantly inhibited PDGF-BB-induced HASMC proliferation and migration. Furthermore, siRNA-SCARA5 significantly inhibited the phosphorylation of PDGF receptor (PDGFR) ß, AKT and extracellular signal-regulated kinase 1/2 in PDGF-BB-stimulated HASMCs. In conclusion, this study demonstrated that knockdown of SCARA5 inhibits PDGF-BB-induced HASMC proliferation and migration through suppression of the PDGF signaling pathway. Thus, SCARA5 may be a novel therapeutic target for preventing or treating vascular diseases involving VSMC proliferation and migration. [ABSTRACT FROM AUTHOR]

The Ras-related C3 botulinum toxin substrate 1 (Rac1)-WASP-family verprolin-homologous protein-2 (WAVE2)-actin-related protein 2/3 (Arp2/3) signaling pathway has been identified to be involved in cell migration and invasion in various types of cancer cell. Cofilin-1 (CFL-1), which is regulated by the Rac1-WAVE2-Arp2/3 signaling pathway, may promote radioresistance in glioma. Therefore, the present study aimed to investigate the potential role of the Rac1-WAVE2-Arp2/3 signaling pathway in radioresistance in U251 human glioma cells and elucidate its affect on CFL-1 expression. Western blot analysis was performed to evaluate the protein expression of CFL-1. In the present study, Rac1 was inhibited by NSC 23766, WAVE2 was inhibited by transfection with short hairpin (sh)RNA-WAVE2 using Lipofectamine? 2000 and Arp2/3 was inhibited by CK-666. Cell viability was measured using the 3-(4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide assay, the cell migration ability was examined by a wound-healing assay, and the cell invasion ability was assessed using a Transwell culture chamber system. The results showed that inhibition of the Rac1-WAVE2-Arp2/3 signaling pathway using NSC 23766, shRNA-WAVE2 or CK-666 reduced the cell viability, migration and invasion abilities in U251 human glioma cells, concordant with a reduced expression of CFL-1. Furthermore, the expression of CFL-1 was significantly increased in radioresistant U251 glioma cells when compared with normal U251 human glioma cells. These findings indicate that inhibition of the Rac1-WAVE2-Arp2/3 signaling pathway may promote radiosensitivity, which may partially result from the downregulation of CFL-1 in U251 human glioma cells. [ABSTRACT FROM AUTHOR]

Emerging data has suggested a high prevalence of osteoarthritis (OA) among obese people. As an important adipokine secreted by white adipose tissue, leptin may be a key mediator in the progression of OA. Leptin exerts a catabolic effect on OA cartilage by increasing the production of metalloproteinase (MMP) enzymes, and contributes to apoptosis in chondrocytes. The current study aimed to explore the role of leptin on the apoptosis of chondrocytes in OA, and its underlying mechanisms. In the in vitro model of OA used in the present study, administration of exogenous leptin induced the generation of reactive oxygen species (ROS) and apoptosis in chondrocytes. It has been demonstrated that leptin is associated with the pathogenesis of OA via the Janus kinase 2 (JAK2)-signal transducer and activator of transcription 3 (STAT3) signaling pathway, and data gathered in the present study demonstrated that suppression of this signaling pathway using a JAK2 inhibitor, AG490, significantly ameliorated leptin-induced apoptosis in damaged chondrocytes in vitro, and reduced the generation of ROS. Furthermore, the protein expression levels of MMP-13 and B-cell lymphoma 2-associated X protein were downregulated in the AG490-treated group. The results of the present study may provide insight into the underlying molecular mechanism by which leptin induces apoptosis in chondrocytes. These findings indicated the importance of leptin as a therapeutic target for the treatment of OA in the overweight population. [ABSTRACT FROM AUTHOR]

Embryonic stem cells (ESCs) are pluripotent stem cells derived from early stage embryos. It remains unclear whether inhibiting the Wnt/β-catenin signaling pathway using dickkopf Wnt signaling pathway inhibitor 1 (DKK1) impacts on the differentiation potential of mouse ESCs in vitro and in vivo. In the present study, immunohistochemical staining was used to measure the expression of markers of the three germ layers in ESCs and teratomas derived from ESCs. The expression of markers for the Wnt/β-catenin signaling pathway were detected by reverse transcription-polymerase chain reaction (RT-qPCR). Immunohistochemistry and western blotting indicated that the expression levels of octamer-binding transcription factor 4 in the DKK1-treated ESC group were significantly greater compared with the control ESCs. Reduced expression levels of NeuroD and bone morphogenetic protein 4 were observed in the DKK1-treated ESCs and teratomas derived from DKK1-treated ESCs compared with the control group. Increased expression levels of SOX17 were observed in the DKK1-treated ESCs compared with the control group. RT-qPCR indicated that β-catenin expression was significantly reduced in DKK1-treated ESCs and teratomas derived from DKK1-treated ESCs compared with the control groups. Western blotting indicated no alterations in the expression of GSK-3β, however, the levels of phosphorylated-GSK-3β were significantly greater in the DKK1 treatment groups, while cyclin D1 and c-Myc expression levels were significantly reduced in the DKK1 treatment groups compared with the control groups. These results suggest that inhibiting Wnt signaling in ESCs using DKK1 may promote mouse ESCs to differentiate into endoderm in vitro and in vivo, and suppress the tumorigenicity of ESCs. [ABSTRACT FROM AUTHOR]

Hepatopulmonary syndrome (HPS) is one of the reasons for the mortality of patients with perioperative liver disease. Intrapulmonary vascular dilatation is the most important mechanism underlying HPS, and it primarily occurs due to cell proliferation. Inhibiting the proliferation of pulmonary microvascular endothelial cells (PMVECs) may provide a novel strategy to prevent HPS. MicroRNAs (miRNAs) regulate gene expression and are crucial in cell proliferation. To investigate the mechanism underlying the proliferation of PMVECs in HPS, PMVECs were isolated from rat models of HPS. It was demonstrated that interleukin (IL)-6 could stimulate the janus kinase 2 (JAK2)/ signal transducer and activator of transcription (STAT3) signaling pathway, which promotes the cell proliferation of PMVECs. JAK2 is a novel target gene of miR-101 and it was shown that miR-101 could inhibit cell proliferation by targeting the IL-6/JAK2/STAT3 signal pathway. In conclusion, the present study demonstrated that miR-101 could inhibit the proliferation of PMVECs by targeting the IL-6/JAK2/STAT3 signaling pathway. This clarifies the role of miR-101 in HPS and provides the theoretical basis of the pathogenesis of HPS. [ABSTRACT FROM AUTHOR]