One-Step Preparation of a Superhydrophobic Surface by Electric Discharge Machining with a Carbon Fiber Brush Electrode.

Superhydrophobic surfaces are extremely susceptible to damage, which can lead to a sharp decrease in their service life and physical properties. Therefore, developing methods to impart superhydrophobic surfaces with excellent wear resistance is crucial. In this article, a flexible carbon fiber brush was utilized as an electrode to fabricate micro-/nano-structures on a grooved surface via electric discharge machining in one step, resulting in a superhydrophobic coating with excellent wear resistance. Carbon fiber brushes exhibit several notable properties, including excellent flexibility, conductivity, and high temperature resistance. Carbon fiber brushes can adapt to the complex inner walls of grooves. Many nano-structures were fabricated on the grooves via pulse discharge, which resulted in a superhydrophobic surface with excellent wear resistance. The contact angle (CA) and sliding angle of the surface after discharge were 156.3 and 2°, respectively. The processed surface exhibits superior corrosion resistance compared to the stainless-steel substrate. The influence of the micro-groove shapes on wear resistance was tested. The results showed that, after 500 cm of wear, the shallow grooves retained their superhydrophobicity with a CA of 150.1°.

One-Step Preparation of Robust Superhydrophobic Foam for Oil/Water Separation by Pulse Electrodeposition.

Frequent leakage and pollution of oily wastewater seriously affect the world's ecosystem safety and economic development, which prompts us to urgently develop a highly effective, low-cost, wear-resistant, chemically stable, and environmentally friendly new functional material for oil/water separation. In this paper, a robust superhydrophobic material was successfully electrodeposited on the porous copper foam substrates in myristic acid (CH3(CH2)12COOH) and lanthanum chloride (LaCl3·6H2O) electrodeposition solution under a continuous pumping circulation and rotation condition. Moreover, SEM, EDS, XRD, FTIR, and XPS technologies were utilized to characterize the surface morphology and chemical composition information. The superhydrophobic property was evaluated by optical contact angle instrument and high-speed camera. It turned out that the micro/nanostructures were mainly composed of lanthanum myristate, and static CA of superhydrophobic copper foam (SCF) was up to 165.2° with SA ≈ 2°. Besides, the SCF exhibited a better performance with good anticorrosion, excellent chemical stability, and outstanding mechanical stability. Furthermore, the SCF can achieve up to 98.6% oil/water separation efficiency. More importantly, by employing this novel processing method, it can effectively save time and provide a promising potential way to make denser and thicker foams for continuous oil/water separation and may be easily applied to other conductive metal matrix materials.

Identification of a novel FGFRL1 MicroRNA target site polymorphism for bone mineral density in meta-analyses of genome-wide association studies.

MicroRNAs (miRNAs) are critical post-transcriptional regulators. Based on a previous genome-wide association (GWA) scan, we conducted a polymorphism in microRNA target sites (poly-miRTS)-centric multistage meta-analysis for lumbar spine (LS)-, total hip (HIP)- and femoral neck (FN)-bone mineral density (BMD). In stage I, 41 102 poly-miRTSs were meta-analyzed in seven cohorts with a genome-wide significance (GWS) α = 0.05/41 102 = 1.22 × 10(-6). By applying α = 5 × 10(-5) (suggestive significance), 11 poly-miRTSs were selected, with FGFRL1 rs4647940 and PRR5 rs3213550 as top signals for FN-BMD (P = 7.67 × 10(-6) and 1.58 × 10(-5)) in gender-combined sample. In stage II in silico replication (two cohorts), FGFRL1 rs4647940 was the only signal marginally replicated for FN-BMD (P = 5.08 × 10(-3)) at α = 0.10/11 = 9.09 × 10(-3). PRR5 rs3213550 was also selected based on biological significance. In stage III de novo genotyping replication (two cohorts), FGFRL1 rs4647940 was the only signal significantly replicated for FN-BMD (P = 7.55 × 10(-6)) at α = 0.05/2 = 0.025 in gender-combined sample. Aggregating three stages, FGFRL1 rs4647940 was the single stage I-discovered and stages II- and III-replicated signal attaining GWS for FN-BMD (P = 8.87 × 10(-12)). Dual-luciferase reporter assays demonstrated that FGFRL1 3' untranslated region harboring rs4647940 appears to be hsa-miR-140-5p's target site. In a zebrafish microinjection experiment, dre-miR-140-5p is shown to exert a dramatic impact on craniofacial skeleton formation. Taken together, we provided functional evidence for a novel FGFRL1 poly-miRTS rs4647940 in a previously known 4p16.3 locus, and experimental and clinical genetics studies have shown both FGFRL1 and hsa-miR-140-5p are important for bone formation.

Serum microRNA-145 as a novel biomarker in human ovarian cancer.

Ovarian cancer is one of the most threatening diseases among women in the world. Current detection methods are expensive and lack accuracy. Thus, a fast, non-invasive biomarker for detecting ovarian cancer is urgently needed. Compelling evidences have been demonstrated that microRNAs, a large family of single-stranded and non-protein-coding RNA molecules, can serve as useful biomarkers in cancer detection. In this study, the relative expressions of microRNA-145 (miR-145) in the serum of patients with ovarian cancer and healthy controls were investigated in an independent study. Subsequently, the diagnosis and prognosis value of miR-145 as a biomarker for ovarian cancer were examined. Furthermore, we performed a meta-analysis to summarize all the results from published studies and this study. Relative expressions of miR-145 were investigated in three independent groups (malignant ovarian cancer, benign ovarian tumor, and healthy controls), comprising a total of 270 participants. Receiver operating characteristic (ROC) curves and overall survival (OS) curves were conducted to compare miR-145 level and clinical characteristics among the three groups. The results showed that relative expressions of the serum miR-145 were significantly down-regulated in patients with malignant ovarian cancer and benign ovarian cancer, compared to healthy controls (P < 0.01). Serum miR-145 levels could discriminate patients with malignant ovarian cancer from healthy controls, with a power area under the curve (AUC) of 0.82 (95 % confidence interval (CI) = 0.77-0.88). Furthermore, patients with low serum levels of miR-145 had a significantly shorter median overall survival rate (hazard ratio (HR) = 1.81, 95 % CI = 1.03-3.17, P = 0.039). The meta-analysis yields good diagnostic performances of miR-145 in various cancers, with an AUC of 0.82 (95 % CI, 0.78-0.85). In conclusion, the present study suggested that miR-145 can potentially serve as an outstanding biomarker for ovarian and other human cancers detection.

Influence of maternal, infant, and collection characteristics on high-quality cord blood units in Guangzhou Cord Blood Bank.

BACKGROUND: The operation of cord blood banks (CBBs) requires immense labor, material, and financial resources. Thus, increasing the ratio of high-quality cord blood units (HQCBUs) in storage that are qualified for clinical use is critical for the efficient use of limited resources. Understanding the factors that contribute to HQCBUs, including maternal, fetal, and processing conditions, may improve the number of HQCBUs in storage. STUDY DESIGN AND METHODS: The maternal, fetal, and processing conditions of 4613 CBUs at the Guangzhou Cord Blood Bank were analyzed retrospectively to determine their effect on HQCBUs. All CBUs were obtained following strict standard operation procedures. RESULTS: Several factors may contribute to HQCBUs: fetal age older than 37 gestational weeks, female fetus, large cord blood (CB) volume (>80 mL), high birthweight (>3500 g), vaginal delivery, and a shorter amount of time between CB collection and processing (12 hr). We report for the first time that α-thalassemia carriers exhibit a postprocessing total nucleated cell count (p-TNCC) increase to at least 1.25 × 10(9) and an increase of the CD34+ cell count to at least 6.01 × 10(6) . Meconium-stained amniotic fluid and mothers younger than 25 years of age exhibited increased p-TNCC to at least 1.25 × 10(9) , and colony-forming units increased to at least 23.24 × 10(5) . CONCLUSIONS: We identified several factors that affect HQCBUs. These results may be used as a reference for updating CB collection strategies, with priority given to collecting CBUs from female fetuses older than 37 gestational weeks, at high birthweight, and born by vaginal delivery from mothers younger than 25 years of age, especially newborns with one parent carrying the trait or with meconium-stained amniotic fluid. The collected CBUs should be sent to the laboratory as soon as possible for priority processing, which will help to increase the number and ratio of HQCBUs and the effective use of CBB resources.

[Application of whole-genome and high-resolution chromosome microarray analysis for the investigation of fetuses with ultrasound abnormalities].

OBJECTIVE: To assess the value of whole-genome high-resolution chromosome microarray analysis (CMA) for the investigation of fetuses with ultrasound abnormalities. METHODS: Whole genome high-resolution CytoScanHD array from Affymetrix was employed to investigate 651 fetuses with structural abnormalities detected by ultrasound, for whom standard G-banded chromosome analysis has revealed a normal karyotype. The fetuses were divided into a single malformation group (n=264) and a multiple malformations group (n=387). In total there were 130 chorionic villus samples, 192 amniotic fluid samples and 329 cord blood samples. Extraction of fetal DNA and CMA experiment have followed the standard guidelines from the manufacturers. All copy number variations (CNVs) detected by CMA were confirmed by fluorescence in situ hybridization (FISH) or real-time polymerase chain reaction (RT-PCR). RESULTS: CMA analysis has detected genomic CNVs in 475 (73%) cases. Clinically significant CNVs were found in 11.5% (75/651) of fetuses, including two uniparental disomies (UPD) and two cryptic mosaicisms. Variations of unknown significance (VOUS) was found in 2.0% (13/651) of tested fetuses. CONCLUSION: Above results have suggested that whole-genome and high-resolution CMA is valuable for the analysis of fetuses with structural abnormalities detected by ultrasound, which can increase the detection rate by approximately 11%. CMA using single nucleotide polymorphism (SNP) array has the ability to detect UPD and low-level mosaicisms. Sufficient communication between technicians and genetic counselors, parental testing and comparison the results with in-house and relevant online databases can significantly reduce the rate of VOUS.

Prenatal diagnosis of congenital heart defect by genome-wide high-resolution SNP array.

OBJECTIVE: This study aimed to detect genomic imbalances in fetuses with congenital heart defect (CHD) by high-resolution single-nucleotide polymorphism (SNP) array. METHODS: A total of 99 fetuses with CHDs with or without other ultrasound anomalies (including structural anomalies and soft markers) but normal karyotypes were investigated using Affymetrix CytoScan HD array. RESULTS: Clinical significant copy number variations (CNVs) were detected in 19 fetuses (19.2%). The proportion for variants of unknown significance was 3% after parental analysis. Five known microdeletion/microduplication syndromes were identified. The detection rate in CHD plus structural anomaly (27.8%) or soft marker (25%) group was higher than but not statistically different from isolated CHD group (15.9%). There was no significant difference between the detection rates in simple and complex CHD groups (20.7% vs. 16.7%). The detection rate in fetuses with CHD and neurologic defect was significantly higher than those with other types of structural anomaly (75% vs. 14.3%, P < 0.05). CONCLUSIONS: Our results demonstrated the value of high-resolution SNP arrays in prenatal diagnosis of CHD; it should become an integral aspect in clinically molecular diagnosis and genetic counseling. The complexity of the cardiac defect was not related to the frequency of clinical significant CNV, but the presence of neurologic defect was related.

KCTD10 is critical for heart and blood vessel development of zebrafish.

KCTD10 is a member of the PDIP1 family, which is highly conserved during evolution, sharing a lot of similarities among human, mouse, and zebrafish. Recently, zebrafish KCTD13 has been identified to play an important role in the early development of brain and autism. However, the specific function of KCTD10 remains to be elucidated. In this study, experiments were carried out to determine the expression pattern of zebrafish KCTD10 mRNA during embryonic development. It was found that KCTD10 is a maternal gene and KCTD10 is of great importance in the shaping of heart and blood vessels. Our data provide direct clues that knockdown of KCTD10 resulted in severe pericardial edema and loss of heart formation indicated by morphological observation and crucial heart markers like amhc, vmhc, and cmlc2. The heart defect caused by KCTD10 is linked to RhoA and PCNA. Flk-1 staining revealed that intersomitic vessels were lost in the trunk, although angioblasts could migrate to the midline. These findings could be helpful to better understand the determinants responsible for the heart and blood vessel defects.

Affinity purification of binding miRNAs for messenger RNA fused with a common tag.

Prediction of microRNA-mRNA interaction typically relies on bioinformatic methods, but these methods only suggest the possibility of microRNA binding and may miss important interactions as well as falsely predict others. A major obstacle to the miRNA research has been the lack of experimental procedures for the identification of miRNA-mRNA interactions. Recently, a few studies have attempted to explore experimental methods to isolate and identify miRNA targets or miRNAs targeting a single gene. Here, we developed an more convenient experimental approach for the isolation and identification of miRNAs targeting a single gene by applying short biotinylated DNA anti-sense oligonucleotides mix to enhanced green fluorescent protein (EGFP) mRNA which was fused to target gene mRNA. This method does not require a design of different anti-sense oligonucleotides to any mRNA. This is a simple and an efficient method to potentially identify miRNAs targeting specific gene mRNA combined with chip screen.

[Analysis of 22 patients with congenital cleft lip and palate using high-resolution chromosome microarray].

OBJECTIVE: To assess the value of chromosome microarray analysis (CMA) for identifying the etiology of patients with congenital cleft lip and palate. METHODS: Twenty-two patients with no identifiable chromosomal aberrations by conventional cytogenetic technique were selected. DNA was extracted and hybridized with Affymetrix CytoScan(TM) HD arrays following the manufacturer's protocol. The data were analyzed with a CHAS v2.0 software. RESULTS: CMA analysis has identified submicroscopic copy number variants (CNVs) in all of the cases, which have ranged from 100 kb to 1.8 Mb. Potential pathogenic CNVs were identified in 5 patients (22.7%), which involved microdeletions and microduplications on 8p23.1, 10q22.2-q22.3, 6q26, 20p12.1 and 18q12.3. MYST4, MACROD2 and SOX7 genes are likely the causative genes. CONCLUSION: CMA is an effective method for identification of etiology in patients with cleft lip and palate. CMA should be provided for patients with cleft lip and palate but a normal karyotype. Especially for those with additional structural abnormalities, there is a high risk for submicroscopic chromosomal aberrations.

Bubble tea consumption and its association with mental health symptoms: An observational cross-sectional study on Chinese young adults.

BACKGROUND: Bubble tea has become enormously popular in China in recent years. This cross-sectional study investigated the association between bubble tea consumption and symptoms of anxiety and depression in a population-based sample. METHODS: Self-report questionnaires were used to assess bubble tea consumption and mental status. Symptoms of depression were assessed using the self-rated Patient Health Questionnaire-2 (PHQ-2) and anxiety using the self-rated Generalized Anxiety Disorder Scale-2 (GAD-2). Logistic regressions were performed to evaluate the association between the frequency of bubble tea consumption (cups/week) (FBTC) and psychological symptoms, adjusting for demographic and socioeconomic characteristics. Path analysis was used to investigate the possible meditating effects of confounding factors. RESULTS: After adjusting for potential confounding factors, high frequency of bubble tea consumption was observed to be significantly associated with high risk for depression in the regression model (p for trend = 0.045); no significant association was found with anxiety (p for trend = 0.332). In path analysis model, education mediated the association between FBTC and depression. Unlike the result in regression model, with education, exercise time and occupation as mediating factors, anxiety was found to be indirectly associated with FBTC. LIMITATIONS: Symptoms of depression and anxiety were self-reported. And the cross-sectional nature of the study does not allow inferring causality. CONCLUSIONS: Bubble tea consumption was associated with an increased risk of experiencing symptoms of depression and anxiety in Chinese young adults. Additional longitudinal research is required to elucidate the possible reverse causation from bubble tea consumption on symptoms of depression and anxiety.

Electric Spark Deposition of Antibacterial Silver Coating on Microstructured Titanium Surfaces with a Novel Flexible Brush Electrode.

Infection caused by orthopedic titanium implants, which results in tissue damage, is a key factor in endosseous implant failure. Given the seriousness of implant infections and the limitations of antibiotic therapy, surface microstructures and antimicrobial silver coatings have emerged as prominent research areas and have displayed certain antimicrobial effects. Researchers are now working to combine the two to produce more effective antimicrobial surfaces. However, building robust and homogeneous coatings on complex microstructured surfaces is a tough task due to the limits of surface modification techniques. In this study, a novel flexible electrode brush (silver brush) instead of a traditional hard electrode was designed with electrical discharge machining, which has the ability to adapt to complex groove interiors. The results showed that the use of flexible electrode brush allowed silver to be deposited uniformly in titanium alloy microgrooves. On the surface of Ag-TC4, a uniformly covered deposit was visible, and it slowly released silver ions into a liquid environment. In vitro bacterial assays showed that a Ag-TC4 microstructured surface reduced bacterial adhesion and bacterial biofilm formation, and the antibacterial activity of Ag-TC4 against Staphylococcus aureus and Escherichia coli was 99.68% ± 0.002 and 99.50% ± 0.007, respectively. This research could lay the groundwork for the study of antimicrobial metal bound to microstructured surfaces and pave the way for future implant surface design.

Secretory carcinoma of the breast with multiple distant metastases in the brain and unfavorable prognosis: a case report and literature review.

BACKGROUND: Secretory carcinoma of the breast is one of the rarest entities, accounting for less than 0.15 % of all infiltrating breast carcinomas. It has characteristic histopathological and molecular features and, in general, a more favorable prognosis. In this case report, we describe a local, advanced secretory carcinoma of the breast with aggressive course and an unfavorable outcome. CASE PRESENTATION: A hard, painless, and palpably bossed mass approximately 12.0 cm in diameter occupied most of the left breast of a 39-year-old woman with fixation to the overlying skin. Breast ultrasonography and magnetic resonance imaging (MRI) scans gave the same grading as BI-RADS IV. A needle biopsy was performed, and the pathological diagnosis was secretory carcinoma. Neoadjuvant chemotherapy (NAC) was then performed, after which ultrasonography and MRI scans revealed chemo-resistance of the tumor to NAC. Left breast mastectomy and axillary lymphadenectomy were subsequently performed. Tumor cells were triple-negative and positive for S-100 and periodic acid-Schiff (PAS) staining. Fluorescence in-situ hybridization (FISH) analysis indicated a fusion arrangement of the ETV6-NTRK3 gene. The patient developed multiple distant metastases in the brain and died of these metastases 19 months after initial diagnosis. CONCLUSIONS: Secretory carcinomas of the breast have been described as a low-grade histologic subtype with a favorable prognosis. This case showed chemo-resistance to neoadjuvant chemotherapy, multiple distant metastases, and a final unfavorable outcome. Further research is needed to better understand the behavior and treatment of this rare tumor.

Upregulated GRB7 promotes proliferation and tumorigenesis of Bladder Cancer via Phospho-AKT Pathway.

Growth factor receptor-bound protein 7 (GRB7) has been found closely related to the occurrence and development of various tumors, but its function in bladder cancer has not yet been elucidated. The study is aiming at investigating the expression and function of GRB7 in bladder cancer. The Cancer Genome Atlas (TCGA) database was selected to analyze mRNA levels of GRB7 in bladder cancer. RT-qPCR and Western blot were conducted to detect the expression of GRB7 in normal bladder epithelial cells, seven bladder cancer cell lines and eight pairs of malignant/nonmalignant bladder tissues. The role of GRB7 in tumor proliferation and tumorigenesis was explored by establishing stable cells, in vitro cell experiments and in vivo xenograft models. The molecular regulation mechanism of GRB7 in bladder cancer was investigated by treatment with AKT inhibitor. GRB7 mRNA was upregulated in bladder cancer samples compared with that in normal tissue samples. Overexpressing GRB7 significantly promoted the proliferation and tumorigenesis of bladder cancer. However, silencing GRB7 played the retarding part. GRB7 promoted G1/S transition by activating the AKT pathway. Our results indicate that GRB7 plays an important role in promoting proliferation and tumorigenesis of bladder cancer.

The Different Effects of IFN-ß and IFN-γ on the Tumor-Suppressive Activity of Human Amniotic Fluid-Derived Mesenchymal Stem Cells.

Current studies have shown that type I or II interferon-modified mesenchymal stem cells have great potential for the application of tumor-targeted therapy, but the underlying mechanism remains largely elusive. Here, we compared the different effects of IFN-ß and IFN-γ on the antitumor activity of human amniotic fluid-derived mesenchymal stem cells (AFMSCs) and revealed the potential mechanism. In detail, AFMSCs primed with IFN-ß or IFN-ß plus IFN-γ, not IFN-γ, inhibited the proliferation of cancer cells in an immunocompetent mouse H460 subcutaneous model, although they all inhibited the proliferation of cancer cells in an immunocompromised mouse H460 subcutaneous model. TRAIL expressed by IFN-ß- or IFN-γ-primed AFMSCs specifically exerted the antitumor effect of AFMSCs. AFMSCs primed with IFN-γ highly expressed immunosuppressive molecule IDO1, but IFN-ß counteracted the IFN-γ-initiated IDO1 expression. 1-MT (IDO1 inhibitor) decreased TRAIL, but increased IDO1 expression in AFMSCs primed with interferon. As a result, AFMSCs primed with IFN-ß or IFN-γ had the antitumor activity, and 1-MT failed to enhance the antitumor effect of IFN-γ-primed AFMSC in vitro and in the immunocompromised mouse H460 subcutaneous model. Furthermore, the expression of TRAIL in AFMSCs was upregulated by apoptotic cancer cells and this positive feedback intensified the antitumor effects of IFNs-primed AFMSCs. The different target gene expression profiles of AFMSCs regulated by IFN-ß and IFN-γ determined the different antitumor effects of IFN-ß- and IFN-γ-primed AFMSCs on tumor cells. Our finding may help to explore a clinical strategy for cancer intervention by understanding the antitumor mechanisms of MSCs and interferon.

Pristimerin overcomes adriamycin resistance in breast cancer cells through suppressing Akt signaling.

Breast cancer remains a major public health problem worldwide. Chemotherapy serves an important role in the treatment of breast cancer. However, resistance to chemotherapeutic agents, in particular, multi-drug resistance (MDR), is a major cause of treatment failure in cancer. Agents that can either enhance the effects of chemotherapeutics or overcome chemoresistance are urgently needed for the treatment of breast cancer. Pristimerin, a quinonemethide triterpenoid compound isolated from Celastraceae and Hippocrateaceae, has been shown to possess antitumor, anti-inflammatory, antioxidant and insecticidal properties. The aim of the present study was to investigate whether pristimerin can override chemoresistance in MCF-7/adriamycin (ADR)-resistant human breast cancer cells. The results demonstrated that pristimerin indeed displayed potent cytocidal effect on multidrug-resistant MCF-7/ADR breast cancer cells, and that these effects occurred through the suppression of Akt signaling, which in turn led to the downregulation of antiapoptotic effectors and increased apoptosis. These findings indicate that use of pristimerin may represent a potentially promising approach for the treatment of ADR-resistant breast cancer.

MicroRNA-140-5p impairs zebrafish embryonic bone development via targeting BMP-2.

MicroRNA-140-5p (miRNA-140-5p) is important for embryonic bone development. In this study, we found that miRNA-140-5p and its binding site in the 3'UTR of bone morphogenetic protein 2 (BMP-2) are highly conserved among vertebrates, and miRNA-140-5p negatively regulates both zebrafish and human BMP-2 genes. Microinjection of miRNA-140-5p or BMP-2b morpholino into zebrafish embryos led to a similar phenotype, including shortened tails, curved trunks, and defects in cranial cartilage. Moreover, miRNA-140-5p injection induced zebrafish embryo malformation that could be significantly rescued by microinjection of BMP-2 mRNA. In conclusion, our results indicated that miRNA-140-5p regulates zebrafish embryonic bone development via targeting BMP-2.

Cdc6 contributes to cisplatin-resistance by activation of ATR-Chk1 pathway in bladder cancer cells.

High activation of DNA damage response is implicated in cisplatin (CDDP) resistance which presents as a serious obstacle for bladder cancer treatment. Cdc6 plays an important role in the malignant progression of tumor. Here, we reported that Cdc6 expression is up-regulated in bladder cancer tissues and is positively correlated to high tumor grade. Cdc6 depletion can attenuate the malignant properties of bladder cancer cells, including DNA replication, migration and invasion. Furthermore, higher levels of chromatin-binding Cdc6 and ATR were detected in CDDP-resistant bladder cancer cells than in the parent bladder cancer cells. Intriguingly, down-regulation of Cdc6 can enhance sensitivity to CDDP both in bladder cancer cells and CDDP-resistant bladder cancer cells. Cdc6 depletion abrogates S phase arrest caused by CDDP, leading to aberrant mitosis by inactivating ATR-Chk1-Cdc25C pathway. Our results indicate that Cdc6 may be a promising target for overcoming CDDP resistance in bladder cancer.

Knockdown of flotillin-2 impairs the proliferation of breast cancer cells through modulation of Akt/FOXO signaling.

Lipid rafts, specialized domains in cell membranes, function as physical platforms for various molecules to coordinate a variety of signal transduction processes. Flotillin-2 (FLOT2), a marker of lipid rafts, is involved in the progression of cancer, yet the precise mechanism remains unclear. In the present study, we examined the effect of FLOT2 on cell proliferation and found that silencing endogenous FLOT2 with shRNAs inhibited proliferation of breast cancer cells. Furthermore, the antiproliferative effect of silencing FLOT2 on breast cancer cells was associated with upregulation of cyclin-dependent kinase (CDK) inhibitors p21Cip1 and p27Kip1. Moreover, we further demonstrated that the silencing of FLOT2 enhanced the transcriptional activity of FOXO factors by decreasing its phosphorylation through inhibiting the PI3K/Akt signaling pathway. Taken together, our results provide the first demonstration of a novel mechanism by which FLOT2 induces proliferation of breast cancer cells, and our findings suggest that FLOT2 plays an important role in oncogenesis of breast cancer and thereby may be a potential target for human breast cancer treatment.

Transcription factor cyclic adenosine monophosphate responsive element binding protein negatively regulates tumor necrosis factor alpha-induced protein 1 expression.

Tumor necrosis factor alpha (TNFα)-induced protein 1 (TNFAIP1) was originally identified as a protein involved in DNA replication, DNA damage repair, apoptosis and the progression of certain diseases, such as Alzheimer's disease. In the present study, forskolin, a stimulant of cyclic adenosine monophosphate (cAMP), was found to significantly reduce human TNFAIP1 mRNA levels and TNFAIP1 promoter activity in the SKNSH human neuroblastoma cell line as indicated by polymerase chain reaction analysis and a luciferase reporter assay. The association between transcription factor cAMP response element­binding protein (CREB) and TNFAIP1 was further investigated using loss- and gain of function-studies with western blot analysis and luciferase reporter assays. The CREB-specific inhibitor KG­501 significantly increased TNFAIP1 protein levels, while overexpression of wild­type CREB, but not CREB mutated at ser133a or its DNA-binding site, significantly decreased human TNFAIP1 protein levels and TNFAIP1 promoter activity in SKNSH cells. Furthermore, two CRE sites located at ­285 and ­425 bp of the human TNFAIP1 promoter were identified to be responsible for CREB­induced inhibition of human TNFAIP1 promoter activity. Chromatin immunoprecipitation assays confirmed that CREB bound to the TNFAIP1 promoter region harboring these two CRE sites. A further luciferase reporter assay demonstrated that CREB phosphorylation on ser133 was responsible for forskolin­induced inhibition of TNFAIP1 expression. In conclusion, the present study suggested that CREB is a negative regulator of the TNFAIP1 gene.