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Hashimoto's thyroiditis (HT) is characterized by dysregulated immune responses and is commonly associated with insulin resistance. However, the mechanism of insulin resistance in HT remains to be fully elucidated. The aim of the present study was to investigate the correlation between the percentage of B regulatory lymphocytes (Bregs) and insulin resistance in patients with HT but with normal thyroid function (type I). A total of 59 patients with type I HT and 38 healthy volunteers were enrolled in the study. An oral glucose tolerance test was performed to measure insulin secretion and assess β‑cell functions. Flow cytometry was performed to examine the percentages of lymphocyte populations. The patients with HT exhibited normal fasting and postprandial glucose and fasting insulin secretion, but increased secretion of early‑phase and total insulin. The patients with HT also had insufficient β‑cell compensation for insulin resistance, indicated by a reduced disposition index, in the fasting state. An elevation in the percentage of CD19+CD24+CD27+ Bregs was also observed, which correlated positively with insulin secretion and insulin resistance in the fasting state. The patients with type I HT had postprandial insulin resistance and insufficient β‑cell compensation for fasting insulin resistance. Therefore, the increase in CD19+CD24+CD27+ Bregs was closely associated with fasting insulin secretion. These results provide novel insight into the mechanism of insulin resistance in HT. [ABSTRACT FROM AUTHOR]

Type 2 diabetes mellitus (T2DM) is characterized by islet β-cell dysfunction and insulin resistance, which leads to an inability to maintain blood glucose homeostasis. Circulating microRNAs (miRNAs) have been suggested as novel biomarkers for T2DM prediction or disease progression. However, miRNAs and their roles in the pathogenesis of T2DM remain to be fully elucidated. In the present study, the serum miRNA expression profiles of T2DM patients in Chinese cohorts were examined. Total RNA was extracted from serum samples of 10 patients with T2DM and five healthy controls, and these was used in reverse-transcription-quantitative polymerase chain reaction analysis with the Exiqon PCR system of 384 serum/plasma miRNAs. A total of seven miRNAs were differentially expressed between the two groups (fold change >3 or <0.33; P<0.05). The serum expression levels of miR-455-5p, miR-454-3p, miR-144-3p and miR-96-5p were higher in patients with T2DM, compared with those of healthy subjects, however, the levels of miR-409-3p, miR-665 and miR-766-3p were lower. Hierarchical cluster analysis indicated that it was possible to separate patients with T2DM and control individuals into their own similar categories by these differential miRNAs. Target prediction showed that 97 T2DM candidate genes were potentially modulated by these seven miRNAs. Kyoto Encyclopedia of Genes and Genomes pathway analysis revealed that 24 pathways were enriched for these genes, and the majority of these pathways were enriched for the targets of induced and repressed miRNAs, among which insulin, adipocytokine and T2DM pathways, and several cancer-associated pathways have been previously associated with T2DM. In conclusion, the present study demonstrated that serum miRNAs may be novel biomarkers for T2DM and provided novel insights into the pathogenesis of T2DM. [ABSTRACT FROM AUTHOR]

The aim of the present study was to investigate whether autophagy is involved in aldosterone (Aldo)-induced mesangial cell (MC) proliferation. MCs were incubated with 10-7 M Aldo for 24 h. Proliferation of MCs, and the underlying mechanisms, were subsequently analyzed using [3H]thymidine assay, cell counting assay, western blotting and RNA interference (RNAi). Aldo was revealed to induce autophagy, as indicated by the increased conversion from microtubule-associated protein 1A/1B-light chain 3 (LC3)-I to LC3-II, the increased expression levels of autophagy-related gene 7 (Atg7) and the increased degradation of p62, which was accompanied by MC proliferation. Notably, pharmacological inhibition of autophagy or RNAi-mediated knockdown of Atg7 attenuated Aldo-induced MC proliferation, suggesting that autophagy was at least partially responsible for this effect. The results of the present study provided evidence that autophagy is critical for regulating Aldo-induced MC proliferation. [ABSTRACT FROM AUTHOR]

Cataract formation is a multifactorial disease, induced by a variety of stressors. The endoplasmic reticulum (ER) stress-induced unfolded protein response (UPR) is known to produce reactive oxygen species (ROS) leading to apoptosis. The present study aimed to investigate whether activation of the UPR occurs in human lenses, using human lens epithelial cell (HLEC) lines and lenses obtained from an eye bank, from individuals aged between 50 and 90 years. In vitro analysis was performed using calcimycin (10 µM) as an ER stressor. The level of ER stress was measured by the production of ROS, staining for cell death, detection of binding immunoglobulin proteins (BIP) and levels of other UPR proteins, including inositol-requiring enzyme-1 (IRE), activating transcription factor (ATF) 6 and PKR-like eukaryotic initiation factor 2a kinase (PERK). These parameters were examined in HLECs exposed to calcimycin for 12, 24, 48 and 72 h. Fluorescent activated cell sorting analysis of the levels of ROS and apoptosis revealed an increase following 24 h calcimycin exposure. The reverse transcription quantitative polymerase chain reaction results demonstrated a gradual increase in the mRNA levels of BIP, IRE1, ATF6 and PERK between 12 and 72 h. A similar effect was observed in the protein levels, which also demonstrated a gradual increase in the levels of endoplasmic oxidoreductin-1-like (Ero1-L)-β and protein disulfide isomerase, but a lower level of Ero1-Lα. Activation of the UPR involved the apoptotic pathway, revealed by increased levels of C/EBP homologous protein, ATF4 and caspase-4. Additionally, the antioxidant protein levels were also suppressed. The investigation of aged human lenses revealed a similar increase in the protein expression of UPR. These results indicated that activation of the UPR-induced ROS production suppressed the antioxidant status and triggered the apoptotic pathway, ultimately leading to the formation of age-related cataracts. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate the effect of the tumor-targeting recombinant human tumor necrosis factor (rhTNF)-α fusion protein mediated by urokinase on Sl80 tumor-bearing mice, as well as to explore its mechanisms of action. Furthermore, the study aimed to observe the effect of the protein on liver and kidney function. rhTNF-α fusion protein prokaryotic expression vectors were constructed using genetic engineering techniques, and were introduced into Escherichia coli. Expression of the fusion protein was induced, and it was then separated and purified in order to determine its cytotoxic activity on L929 cells. Kunming mice were randomly divided into four groups after being inoculated with S180 tumor cells. The groups were then injected with saline (control group, group S), or saline with 0.1 μg/ml fusion protein (low dose group, group L), 0.2 μg/ml fusion protein (middle dose group, group M) or 0.3 μg/ml (high dose group, group H). The mice were sacrificed after 12 days and liver [mg/kg; (liver weight/body weight) × 1,000] and kidney [mg/kg; (kidney weight/body weight) × 1,000] indices, tumor weight, the percentage reduction in mean tumor size, and the levels of alanine transaminase (ALT), albumin (ALB), creatinine (Cr) and blood urea nitrogen (BUN) in each group of mice were determined. In addition, the levels of urokinase-type plasminogen activator (uPA), the expression of bcl-2, bax and vascular endothelial growth factor (VEGF), and the percentage of apoptotic cells were measured with an enzyme-linked immunosorbent assay, streptavidin-biotin complex of immunohistochemistry and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling, respectively. The fusion protein significantly inhibited the growth of S180 tumor cells in vivo in a dose-dependent manner. With an increase in the dose of fusion protein, ALT, uPA, bcl-2 and VEGF levels decreased, and ALB levels increased. However, liver and kidney indices and bax expression were not significantly altered. Cr and BUN levels did not change significantly in the low and middle dose groups, but did increase in the high dose group. Compared with the control group, the percentage of apoptotic cells in the high-dose group was significantly higher. In conclusion, the fusion protein significantly inhibited S180 tumor growth in a mouse model, possibly by reducing the levels of uPA, bcl-2 and VEGF. There was a mildly toxic effect on the kidneys with the high dose, but a protective effect in the liver. [ABSTRACT FROM AUTHOR]

Cell differentiation agent II (CDA-II) is a DNA methyltransferase inhibitor isolated from healthy human urine. In the present study, the antitumor activity of CDA-II on human multiple myeloma (MM) cell lines via the mitochondrial pathway was first revealed. The human MM cell lines were exposed to CDA-II. Cytotoxicity, caspase activation, apoptosis and the effects on the mitochondrial pathway were assessed. CDA-II was capable of decreasing the depolarized mitochondrial membranes and activating caspase 3 and 9 and poly (ADP ribose) polymerase in MM cells treated with CDA-II. CDA-II induced caspase dependent cell death accompanied by a significant decrease in X-linked inhibitor of apoptosis protein (XIAP), survivin and Mcl 1 levels. The caspase 3 inhibitor, Z DEVD FMK, inhibited CDA-II induced apoptosis. CDA-II potently increased the Bax levels, decreased the Bcl 2/Bax ratio and decreased the expression of the downstream targets of NF κB. In conclusion, the results of the present study demonstrated that CDA-II treatment leads to the inhibition of p65 nuclear localization and potently induces caspase dependent apoptosis in MM cells mediated through the mitochondrial pathway at low nanomolar concentrations. These results indicate that CDA-II is a novel inhibitor of NF κB activity, with notable antimyeloma efficacy. This study provides a rationale for the clinical investigation of CDA-II in human MM. [ABSTRACT FROM AUTHOR]

Differentially-expressed genes (DEGs) correlated to oral contraceptives (OCs) were identified by comparing the transcriptomes of cervical cancer patients who have taken OCs and those who have not. Their biological functions and relevance to clinical manifestations were investigated further in order to gain an understanding of the pathogenesis of cervical cancer and provide potential therapeutic targets. Level 3 RNA-sequencing (seq) data for cervical squamous-cell carcinoma and endocervical adenocarcinoma and the clinical information were downloaded from The Cancer Genome Atlas. The present study analyzed the RNA-seq data and information on OC use of 35 patients [OC users (n=18) and those who have never used OCs (n=17)]. Student's t-test was used in order to identify DEGs and the false discovery rate (FDR) was estimated by a Beta-Uniform Mixture model, which was adopted in multiple testing corrections. A functional enrichment analysis was performed with the Database for Annotation, Visualization and Integrated Discovery tool and BioCarta. A total of 80 DEGs were identified in OC users while FDR=0.3 was set as the cut-off value. The metabolic process and human telomerase RNA gene transcription were significantly upregulated in DEGs. Furthermore, secreted LY6/PLAUR domain containing 1 was identified to be correlated to the pathological response, while the synapse defective 1 Rho GTPase homolog 2 was found to be significantly associated with the histological grade and overall survival time. In conclusion, present study shed light on the effect of OC use on the oncogenesis of the cervix and may indicate novel approaches for a targeted therapy of cervical cancer. [ABSTRACT FROM AUTHOR]

A number of short noncoding microRNAs (miRs) have been demonstrated to be highly expressed in many kidney diseases such as renal cancer and lupus nephritis (LN); however, these results have not been extensively investigated. The aim of the present study was to investigate the expression and function of miR‑198 in LN based on the previous studies. miR‑198 expression level in systemic lupus erythematosus (SLE) patients was determined to determine its clinicopathological significance and effect on glomerular cell proliferation. It was demonstrated that higher expression of miR‑198 was observed in patients with SLE, and was correlated with disease activity. Bioinformatics prediction and luciferase assays were used to demonstrate that miR‑198 could directly bind to the phosphatase and tensin homology deleted on chromosome ten (PTEN) 3'‑untranslated region. Furthermore, miR‑198 overexpression reduced PTEN expression levels, while miR‑198 silencing increased its expression at both the mRNA and protein level. Furthermore, there was a negative association between miR‑198 and PTEN in the patients with active SLE. Thus, miR‑198 may promote proliferation and contribute to SLE progression by targeting PTEN. [ABSTRACT FROM AUTHOR]

The present study aimed to investigate the effects of Staphylococcus aureus enterotoxin C3 (SEC3), including recombinant (r)SEC3 protein and lentivirus‑mediated SEC3, on the activation, proliferation and cytokine production of human T cells. HeLa cells were infected with SEC3 lentiviral vector (LV‑SEC3) and viability was determined using the Cell Counting Kit‑8 (CCK‑8) assay. Subsequently, infected cells or rSEC3 protein were co‑cultured with human peripheral blood mononuclear cells (PBMCs) for 10 days, after which the culture supernatant and T cells were incubated with untreated HeLa cells, which were subjected to a CCK‑8 assay to determine cytotoxicity. In addition, IL‑6 and IFN‑γ expression was detected by chemiluminescence and enzyme‑linked immunospot analyses, respectively. Subpopulations of activated T cells were sorted by flow cytometry. The results demonstrated that, following infection with LV‑SEC3 or negative control lentiviral vector (LV‑NC), >80% of HeLa cells presented green fluorescent protein‑positive signals. All five groups of co‑cultured T cells exhibited proliferation. Co‑culture of PBMCs with rSEC3 protein or LV‑SEC‑infected cells resulted in elevated IL‑6 and IFN‑γ secretion. In addition, rSEC3‑activated and monocultured T cells were predominantly cluster of differentiation (CD)4+ (62.7 and 59.6%, respectively) whereas phytohemagglutinin‑stimulated T cells were predominantly CD8+ (57.8%). Compared with the LV‑NC group, T cells and culture supernatants from the LV‑SEC3 group significantly attenuated proliferation of HeLa cells. These results suggest that rSEC3 protein, and LV‑SEC3‑infected HeLa cells, are able to potently activate T cells, increasing cytokine production and amplify the antitumor immune response. [ABSTRACT FROM AUTHOR]

Aldosterone (ALD) is a well-known hormone, which may initiate renal injury by inducing mesangial cell (MC) injury in chronic kidney disease (CKD); however, the molecular mechanism remains unknown. The aim of the present study was to investigate the effects of p53 on ALD-induced MC apoptosis and elucidate the underlying molecular mechanism. For the in vivo studies, rats were randomly assigned to receive normal saline or ALD for 4 weeks. The ratio of MC apoptosis was analysed by terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) assay. In addition, the expression level and localisation of p53, a well-known cell apoptosis-associated key protein, were detected by immunofluorescence. For the in vitro studies, rat MCs were incubated in medium containing either buffer (control) or ALD (10-6 M) for 24 h. The cell apoptosis ratio was assessed by flow cytometry, and the expression level of p53 was assessed by reverse transcription quantitative polymerase chain reaction and western blotting. In order to confirm the role of p53 in ALD-regulated cell apoptosis, a rescue experiment was performed using targeted small interfering (si)RNA to downregulate the expression of p53. The ALD-treated rats exhibited greater numbers of TUNEL-positive MCs and higher expression levels of p53 when compared with the control group. Furthermore, the ratio of MC apoptosis and the p53 expression level were significantly increased following ALD exposure, compared with the control group. Additionally, in the rescue experiment, the effects of ALD on MC were blocked by downregulating the expression level of p53 in MCs. The present study hypothesized that ALD may directly contribute to the occurrence of MC apoptosis via p53, which may participate in ALD-induced renal injury. [ABSTRACT FROM AUTHOR]

MicroRNAs are identified as negative regulators in gene expression through silencing gene expression at the post-transcriptional and translational levels. Bone morphogenetic protein 9 (BMP9) is the most effective in inducing osteogenesis in the BMP family, the members of which were originally identified as osteoinductive cytokines. In the current study, the role of miR-23b in the progression of BMP9-induced C2C12 myoblasts was investigated. The results indicated that miR-23b was significantly downregulated in C2C12 myoblasts induced by BMP9. Overexpression of miR-23b significantly inhibited osteogenesis in the C2C12 myoblasts. In addition, it was observed that Runx2 was negatively regulated by miR-23b at the post-transcriptional level, via a specific target site within the 3'UTR of Runx2. Knockdown of Runx2 promoted miR-23b-induced inhibition of osteogenesis in C2C12 myoblasts. The expression of Runx2 was observed to be frequently upregulated in osteoblast cell lines and inversely correlated with miR-23b expression. Thus, the results of the present study suggest that miR-23b inhibits BMP9-induced C2C12 myoblast osteogenesis via targeting of the Runx2 gene, acting as a suppressor. The current study contributes to the understanding of the functions of BMP9 in ossification. [ABSTRACT FROM AUTHOR]

Cancer is one of the most eminent diseases of modern times and numerous natural products derived from medicinal plants have been identified as potential sources of antitumor drugs. A successful anticancer drug must target or inhibit tumor cells whilst causing minimal damage to healthy cells. The present study aimed to investigate the antitumor efficacy of ethyl acetate extract, and other isolated compounds from Artemisia indica, on MCF-7, BHY, Miapaca-2, Colo-205 and A-549 cell lines. The apoptotic activity of the compounds was studied using flow cytometry. The different cancer cell lines were treated with the ethyl acetate extract and varying concentrations of compounds (denoted a-g) isolated from the A. indica. The cytotoxicity was evaluated by MTT assay and the apoptotic properties of the compounds and the extract were assessed using flow cytometry. In MCF-7 cells, the effect on mitochondrial membrane potential loss (ΛΨm) induced by compounds b and d was also studied. Bioassay-guided fractionation of the ethyl acetate extract from the shoot and root parts of A. indica led to the identification of the compounds a-g as: 5-hydroxy-3,7,4'-trimethoxyflavone; ludartin; maackiain; lupeol; cis-matricaria ester; trans-matricaria ester; and 6-methoxy-7,8-methylenedioxy coumarin, respectively. All the compounds exhibited mild to potent inhibition of cell proliferation in all the cell lines, with the half maximal inhibitory concentration values ranging from 25.18-88.12 μM. Ludartin and lupeol were observed to have the most potent inhibitory effects. Based on the initially identified antiproliferative effects, these two compounds were evaluated for their effects on cell cycle phase distribution, DNA damage and their effects on mitochondrial membrane potential loss (ΛΨm). The two compounds induced DNA damage and mitochondrial membrane potential loss in MCF-7 cells. The results of the current study suggest that lupeol and ludartin, isolated from A. indica, produce anticancer effects by inducing DNA damage and a reduction of mitochondrial membrane potential, and may be used as potent anticancer agents, subsequent to further study. [ABSTRACT FROM AUTHOR]

Subarachnoid hemorrhage (SAH), one of the serious types of stroke incurred by bleeding into the space surrounding the brain, occurs when brains are deprived of oxygen by various factors, particularly an interruption to the blood supply or a ruptured aneurysm. Cerebral vasospasm (CVS) is one of the most common complications of SAH. It has been proposed that platelet-derived growth factor (PDGF) is involved in CVS. The aim of the present study was to analyze expression of PDGF in rabbit models of CVS. Post-SAH CVS rabbit models were created using endovascular puncture and employed to analyze the expression patterns of PDGF by enzyme-linked immunosorbent assay and immunohistochemistry. The results indicated that the creation of the rabbit model of CVS induced using endovascular puncture was successful and demonstrated the double phase changes observed in human CVS. The acute stage started at 12 h post-SAH with narrowing of the vascular lumen diameter. This narrowing appeared again on the seventh day in delayed CVS alongside increased thickness of vessel walls. PDGF-β expression was observed in vascular smooth muscle cells of the rabbit models. PDGF-β was expressed as early as 3 h post-SAH, it was evident after 1 day and reached a peak in 7 days, suggesting that PDGF-β is involved in the early stages of CVS. In the current study, it was confirmed that PDGF-β expression was present in the rabbit models of CVS, which may aid the elucidation of the pathogenesis of CVS, and also provide useful information for diagnosis and treatment of CVS. [ABSTRACT FROM AUTHOR]

Pulmonary artery hypertension (PAH) is a disease with high morbidity and mortality. Cyanidin-3-O-β-glucoside (Cy-3-g), a classical anthocyanin, has a variety of biological effects. The present study evaluated whether Cy-3-g attenuated PAH, and explored the potential mechanism of action. Rats were injected with monocrotaline (MCT; 60 mg per kg of body weight) and then treated with Cy-3-g (200 or 400 mg per kg of body weight) for 4 weeks. Protein expression was determined in vitro in transforming growth factor-β1 (TGF-β1)-mediated human pulmonary arterial smooth muscle cells (SMCs). The results indicated that Cy-3-g significantly inhibited the mean pulmonary artery pressure, right ventricular systolic pressure and right ventricular hypertrophy index, as well as vascular remodeling induced by MCT in PAH rats. Further experiments showed that Cy-3-g suppressed the expression of pro-inflammatory factors and enhanced the levels of anti-inflammatory factors. Cy-3-g blocked oxidative stress and improved vascular endothelial injury. Cy-3-g also reduced the proliferation of SMCs. Furthermore, the MCT- and TGF-β1-induced increase in TGF-β1, phosphorylated (p)-p38 mitogen-activated protein kinase (MAPK) and p-cAMP-response element binding protein (CREB) expression was blocked by Cy-3-g treatment in vivo and in vitro. These results indicated that Cy-3-g could prevent vascular remodeling in PAH via inhibition of the TGF-β1/p38 MAPK/CREB axis. [ABSTRACT FROM AUTHOR]