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Object To identify the expression levels of ECT2 (epithelial cell transforming sequence 2) in triple-negative breast cancer (TNBC) before and after administration of paclitaxel (PTX) and explore the interaction between ECT2 and PTX in breast cancer treatment. Methods Lentiviral (LV) packaging ECT2 overexpression and interference plasmids were constructed for in vitro assays. The effects of ECT2 expression on the TNBC cell line (HCC1806), particularly its roles in the proliferation, invasion, migration and apoptosis and cell cycle, were evaluated using the CCK-8 and other methods before and after PTX treatment. In nude mouse xenograft settings were performed to detect cell apoptosis and Ki-67 expression levels by TUNEL and immunohistochemical staining, respectively. Results In the vitro assays, before and after the PTX treatment, comparison of the LV-ECT2 and sh-ECT2 groups and the remaining three groups (control, LV-NC, sh-NC) showed statistically significant differences in terms of cell proliferation, invasion and migration and apoptosis and changes in the cell cycle. In the vivo assays, the control, LV-ECT2 and sh-ECT2 groups markedly outweighed the corresponding PTX-treated groups. The LV-ECT2, PTX, sh-ECT2 and sh-ECT2-PTX were all significantly different from the control group in terms of body weight and tumour size changes. Cell apoptosis occurred in the PTX, sh-ECT2 and sh-ECT2-PTX groups. About the Ki-67 proliferation index, the PTX, LV-ECT2-PTX, sh-ECT2 and sh-ECT2-PTX groups were significantly different from the control group. Conclusion ECT2, which is a major driving factor in the growth of breast cancer cells, plays an important role in regulating TNBC growth. PTX therapy had significantly improved efficacy after silencing ECT2. This finding indicates that the inhibition of ECT2 expression may facilitate the treatment of breast cancer as a new regimen and provide a theoretical basis for the development of new targeted drugs as a replacement for PTX in breast cancer treatment.

Infectious diseases are a long-standing and severe global public health problem. The rapid diagnosis of infectious diseases is an urgent need to solve this problem. MicroRNA (miRNA) plays an important role in the intervention of some infectious diseases and is expected to become a potential biomarker for the diagnosis and prognosis of infectious diseases. It is of great significance to develop rapid and sensitive methods for detecting miRNA for effective control of infectious diseases. In this study, a simple and highly sensitive homogeneous electrochemical method for microRNAs using enzyme-driven cascaded signal amplification has been developed. In the presence of target miRNA, the reaction system produced plenty of MB-labeled single-nucleotide fragments (MB-MF) containing a few negative charges, which can diffuse to the negative surface of the ITO electrode easily, so an obvious electrochemical signal enhancement was obtained. Without the target, MB-HP contains a relatively large amount of negative charges due to the phosphates on the DNA chain, which cannot be digested by the enzyme and cannot diffuse freely to the negatively charged ITO electrode, so only a small signal was detected. The enhanced electrochemical response has a linear relationship with the logarithm of miRNA concentration in the range of 10 fM to 10 nM and the limit of detection as low as 3.0 fM. Furthermore, the proposed strategy showed the capability of discriminating single-base mismatch and performed eligibly in the analysis of miRNA in cell lysates, exhibiting great potential for disease diagnosis and biomedical research. Graphical abstract.

Vacuum brazing of TiC/Ti matrix composite was performed using Ti-28mass%Ni eutectic brazing alloy. The brazed joints were obtained at different temperatures and holding times. The microstructure and mechanical properties of the joints were investigated in detail. The results show that the typical interfacial microstructure of the brazed joints is Ti solid solution, δ-Ti 2 Ni phase and TiC particles. As the brazing temperature was increased from 1283 K/10min to 1353 K/10min, the joint shear strength first increased and then decreased. Prolonging the holding time, caused the shear strength of the joints to decrease. The maximum shear strength reached 711.4 MPa at 1333 K for 10 min, which is more than twice that obtained with the conventional brazing of Ti alloys. The fracture location varied from the brazing joints to the substrate in different brazing processes. The fracture path, located at the substrate, can only be found at 1333 K/10min. The microstructural evolution of the brazed joints with different brazing processes was clarified and its relationship with the mechanical performance was revealed in detail.

A sensitive and homogeneous electrochemical aptasensor was fabricated for the detection of mucin 1 (MUC1) by combining a well-designed DNA bulge-loop (L-DNA) structure with high-efficient exonuclease I (Exo I)-assisted target recycling amplification strategy. The L-DNA probe was constructed via the hybridization of the MUC1 aptamer and methylene blue (MB) labeled complementary DNA (cDNA) (cDNA-MB) and hence could not diffuse freely to the negatively charged ITO electrode surface due to the strong electrostatic repulsion, so small electrochemical signal was detected. The addition of MUC1 caused the dissociation of L-DNA structure due to the specificity between aptamer and MUC1. Then Exo I was implemented to digest the released cDNA-MB into mononucleotides and then produced short MB-labeled mononucleotides fragments (MB-MFs). As the MB-MFs contained few negative charges, it diffused easily to the negatively charged ITO electrode surface and resulted in the enhanced electrochemical signal. Meanwhile, the MUC1-aptamer complex was also specifically digested by Exo I, resulting in the liberation of MUC1 and hence realized the target recycling and then caused the amplification of the electrochemical signal. The enhanced electrochemical signal has a good linear relationship with logarithm of MUC1 concentration in the range of 1.0 pg mL−1–50 ng mL−1 with a limit of detection of 0.40 pg mL−1 (S/N = 3). Additionally, the fabricated aptasensor has been successfully applied to detect MUC1 in serum samples with satisfactory results and thereby it exhibits great potential in the practical application of clinical diagnosis.

A simple, low-cost, and sensitive liposome-based colorimetric aptasensor has been developed to detect ochratoxin A (OTA). Specifically, a dumbbell-shaped probe was designed, including magnetic beads (MBs), double-stranded DNA (dsDNA), and enzyme-encapsulated liposome. The dsDNA formed by the hybridization between OTA aptamer and its complementary probe. And the dsDNA was used to contact the MBs and the enzyme-encapsulated liposome. In the presence of OTA, the aptamer preferred to combine with OTA to form G-quadruplex, resulting in the release of the detection probe and the enzyme-encapsulated liposome. Each liposome contained a large amount of HRP. Thus, when the liposome was lysed by adding the mixed solution of 3,3′,5,5′-tetramethylbenzidine (TMB) and H2O2, a large number of HRP were released. HRP could catalyze the H2O2-mediated oxidation of TMB and hence resulted in the color change from colorless to blue with the OTA concentration varying, and this variation can be observed by naked eyes easily. The result showed that the absorption intensity at 652 nm enhanced with the increase of OTA concentration ranging from 0.05 to 2.0 ng mL−1, and the limit of detection was calculated to be 0.023 ng mL−1 (S/N = 3). The developed colorimetric aptasensor has been applied to detect OTA concentration in corn samples with satisfied results.

The aim of the present study was to expound on the interactions between the mitogen-activated protein kinase (MAPK) and Hippo pathway members, and to further elucidate the molecular mechanisms of melanoma tumorigenesis. Four melanoma cell lines (C32, HS695T, SK-MEL-28 and A375) were used in the present study. Western blotting was used to assess the expression levels of the MAPK and Hippo pathway effector proteins: rapidly accelerated fibrosarcoma-1 proto-oncogene, serine/threonine kinase (RAF-1); serine/threonine kinase 3 (STK3; also known as MST-2); yes-associated protein (YAP); and tafazzin (TAZ). Immunoprecipitation was used to identify interactions between the effector proteins of the Hippo and MAPK pathways. RAF-1 was knocked down in melanoma cells using siRNA transfection, and cell proliferation, migration and invasion were determined by the MTT, wound-healing and Transwell invasion assays, respectively. Additionally, the cell cycle and apoptosis were analyzed by flow cytometry 48 h after RAF-1 knockdown. We found that the expression levels of the four proteins were variable, and that the HS695T cells expressed the highest levels of RAF-1. Immunoprecipitation studies revealed that RAF-1 bound to MST-2 in melanoma cells. Knockdown of RAF-1 inhibited the expression of YAP and TAZ, but did not affect MST-2 expression. Additionally, RAF-1 knockdown in melanoma cells significantly inhibited cell proliferation, migration and invasion, and induced apoptosis in these cells. Collectively, our results indicate that the RAF-1/MST-2 interaction may be a novel link between the MAPK and Hippo pathways.

A glycoside prodrug of 4-aminosalicylic acid (4-ASA) with d-glucose was synthesized for targeted drug delivery to inflammatory bowel. The in vitro assessment of 4-aminosalicylic acid-β-O-glucoside (4-ASA-Glu) as a colon-specific prodrug was studied using colitis rat with the healthy one as control. The stability studies in aqueous buffers (pH 1.2, 6.8 and 7.4) indicated that 4-ASA-Glu was stable over a period of 12 h. The incubation of 4-ASA-Glu with cecal or colonic contents of healthy rats at 37 °C released 4-ASA in 77 or 80% of the dose in 12 h, respectively. The amount of 4-ASA liberated from the incubation of 4-ASA-Glu in cecal or colonic contents of colitis rats at 37 °C was 69 or 79% in 12 h respectively, while less than 9% 4-ASA was detected from the incubation of 4-ASA-Glu with the homogenates of stomach or small intestine. The curative effect of 4-ASA-Glu was evaluated in 2, 4, 6-trinitrobenzenesulfonic acid (TNBS) induced experimental colitis model in male Sprague-Dawley (SD) rats. It was found that 4-ASA-Glu possess significantly ameliorate effect than sulfasalazine, oral 4- and 5-aminosalicylic acid.

AimsTo investigate the expression of epithelial cell transforming sequence 2 (ECT2) in invasive breast cancer and its prognostic significance.MethodsECT2 immunohistochemical detection was performed in 165 breast cancer specimens and 100 normal control tissues. Univariable and multivariable Cox proportional hazards regression model analysis was used to confirm independent prognostic factors. The PHREG procedure linear hypotheses testing method was used to analyse survival data.ResultsExpression of ECT2 in breast cancer was significantly higher than that of the normal control group (p0.05). Univariable analysis showed that expression of ECT2, the status of lymph node metastasis, triple-negative disease and Ki-67 proliferation index were related to the overall survival of patients with breast cancer (pConclusionsExpression of ECT2 may be one of the main causes of the occurrence and development of breast cancer, and high expression of ECT2 as an independent prognostic factor predicts a poor prognosis. ECT2 could also be a potential molecular target for designing therapeutic strategies for breast cancer.

Objective Yes-associated protein (YAP) was defined as a candidate oncogene in multiple cancers. Yet, the role of YAP in cervical cancer is largely unknown. The aim of this study was to determine whether YAP could be used as a predictive biomarker in cervical precancerous lesions. Methods Immunohistochemical analysis of YAP expression was performed in 10 chronic cervicitis, 49 cervical intraepithelial neoplasia (CIN) 1, 55 CIN 2, 34 CIN 3, and 32 cervical squamous cell carcinoma (SCC) samples. Human papillomavirus (HPV) was detected by HPV genotype detection kit in 70 cases including 10 chronic cervicitis cases, 13 CIN 1 cases, 19 CIN 2 cases, 14 CIN 3 cases, and 14 SCC cases. Furthermore, the relationship between YAP expression and HPV integration status was analyzed using Spearman rank correlation coefficient test. Result Samples of chronic cervicitis had negative or weak expression of YAP in cytoplasm. In the CIN 1 group, YAP expression was primarily confined to the lower third part of squamous epithelia or basal layer, whereas higher-grade CIN (2 and 3) and SCC groups had a strong nuclear expression of YAP. The expression levels of YAP in chronic cervicitis and CIN 1 were significantly lower compared to those in higher-grade CIN and SCC. Moreover, YAP expression was correlated with HPV integration status. Most high-risk HPV(+)/YAP(+) cases were found in the CIN 3 and SCC groups. Conclusion This study suggested that YAP could function as a predictive marker in CIN and cervical cancer. YAP expression, in combination of HPV, might facilitate the identification of precancerous cervical lesions.

It can sometimes be difficult to distinguish between the 2 main types of uterine mesenchymal neoplasms; uterine smooth muscle tumors (SMTs) and endometrial stromal sarcomas (ESSs), particularly when the ESSs show smooth muscle differentiation or the SMTs are highly cellular. The aim of this study was to investigate myocardin expression in normal uterus myometrium, in SMTs, and in ESSs and to determine whether myocardin can be used as a useful diagnostic tool in the classification of problematic uterine mesenchymal tumors. Immunohistochemical staining was performed in each group. Besides myocardin, all cases were also stained for other smooth muscle markers (h-caldesmon, desmin, smooth muscle actin) and for CD10. All tested markers were analyzed in 21 conventional leiomyomas (LMs), 21 highly cellular leiomyomas (HCLs), 12 leiomyosarcomas (LMSs), 3 endometrial stromal nodules (ESNs), 11 ESSs, and 15 normal uterus myometrium. Myocardin was expressed in all normal uterine myometrium and in SMTs (even in the regions with epithelioid features) moderately or strongly, at least topically, whereas in endometrium, in ESNs and in ESSs, except in the regions of smooth muscle differentiation, it was negative. All ESNs, 11 of 11 ESSs and 14 of 15 endometrium were negative for h-caldesmon, but all SMTs and normal uterine myometrium were positive for h-caldesmon except for 2 LMSs, 2 HCLs, and for the regions with epithelioid features in 2 LMs. Desmin was stained in all normal uterine myometrium and in SMTs (except those of the regions with epithelioid features), but it was negative in 1 HCL and 1 LMS. One of 3 ESNs and 2 of 11 ESSs were expressed in desmin. Smooth muscle actin was negative in all ESNs, 2 LMSs, and 2 HCLs, and positive in all myometriums, LMs (except for the regions with epithelioid features), 1 ESSs, and 1 proliferative phase endometrium. Eight of 11 ESSs and all ESNs were positive for CD10, as was 1 HCL, and 2 LMSs. All uterine myometrium, 3 ESSs, and 3 endometriums were negative for CD10. Our study indicates that the myocardin is expressed in normal and neoplastic uterine smooth muscle cells sensitively and that the evaluation of myocardin expression is useful in distinguishing SMTs from ESSs.

Aims: To detect microsatellite loci alterations by fluorescent multiplex PCR in urine sediment cell of urothelial carcinoma, and to determine if they can be used as genetic markers for diagnosis of urothelial carcinoma. Materials and Methods: Microsatellite alteration analysis was conducted using fluorescent multiplex PCR with samples from 64 cases of urothelial carcinomas of the bladder. Three microsatellites spanning the 3p14 and two additional microsatellites in 9q33 and 9p22 were analyzed. Microsatellite alterations (microsatellite instability and/or loss of heterozygosity) in urine sediment cells of urothelial carcinoma patients matched for peripheral blood and tumor tissue were all analyzed. Results: The frequency of microsatellite alterations in urothelial carcinoma was chromosome 3p (D3S1234: 14.6% (7/48), D3S1300: 16.7% (8/48), D3S1313: 8.35% (4/48)); 9q (D9S242: 33.3% (16/48)), and 9p (D9S162: 27.1% (13/48)). Microsatellite alterations happened in 62.5% (40/64) of the patients when combined with all five markers. Our study showed a significant correlation between the microsatellite alteration of the five-locus panel and recurrence (p = 0.010) and smoking habit (p = 0.006). Conclusions: The results suggest that these microsatellite loci alterations may have an important role in the recurrence of urothelial carcinomas. Further studies are needed to better determine the effect of microsatellite loci alterations on prognosis.

AIM: To explore the inhibitory effects of dobutamine on gastric adenocarcinoma cells. METHODS: Dobutamine was used to treat gastric adenocarcinoma cells (SGC-7901) and cell viability was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The effects of dobutamine combined with cisplatin on cell viability were also analyzed. Cell migration was studied using the wound healing assay, and cell proliferation was analyzed using the colony formation assay. A cell invasion assay was carried out using Transwell cell culture chambers. The cell cycle and cell apoptosis were analyzed by flow cytometry. Western blot and immunocytochemistry were performed to determine the expression of Yes-associated protein (YAP) in treated cells. RESULTS: Dobutamine significantly inhibited cell growth, migration, cell colony formation, and cell invasion into Matrigel. Dobutamine also arrested the cell cycle at G1/S phase, and increased the rate of apoptosis of gastric adenocarcinoma cells. The expression of YAP was detected mainly in the nucleus in the absence of dobutamine. However, reduced expression of phosphorylated YAP was mainly found in the cytosol following treatment with dobutamine. CONCLUSION: Dobutamine has significant inhibitory effects on gastric adenocarcinoma cells and may be used in neoadjuvant therapy not only for gastric cancer, but also for other tumors.

Background SPARC (secreted protein, acidic and rich in cysteine) is closely related with the progress, invasion and metastasis of malignant tumor and angiogenesis. Methods Using human colon adenocarcinoma tissues (hereinafter referred to as colon cancer) and their corresponding non-diseased colon from 114 patients' biopsies, the expression of SPARC and vascular endothelial growth factor (VEGF) were investigated by immunohistochemistry staining to assessment the relationship between SPARC and VEGF, as well as their prognostic significance in patients. Evaluation of VEGF expression level with the same tissues was used to establish the antigenic profiles, and the marker of CD34 staining was used as an indicator of microvessel density (MVD). Results SPARC expression was mainly in the stromal cells surrounding the colon cancer, and was significant difference in those tissues with the lymph node metastasis and differentiation degree of tumor. Expression of SPARC was significantly correlated with the expression of VEGF and MVD in colon cancer tissues. Patients with low or absence expressing SPARC had significantly worse overall survival and disease-free survival in a Single Factor Analysis; Cox Regression Analysis, SPARC emerged as an overall survival and disease-free survival independent prognostic factor for colon cancer. Conclusion The low expression or absence of stromal SPARC was an independent prognostic factor for poor prognosis of colon cancer. SPARC maybe involved in the regulation of anti-angiogenesis by which it may serve as a novel target for colon cancer treatment as well as a novel distinctive marker.