LncRNA MHENCR Predicts Poor Outcomes in Patients with Colorectal Carcinoma and Modulates Tumorigenesis by Impairing MiR-532-3p.

The long noncoding RNAs (lncRNAs) are widely involved in the progression of various malignant tumors. The current study investigated the role and mechanism of lncRNA melanoma highly expressed noncoding RNA (MHENCR) in colorectal carcinoma. The expression of MHENCR was measured by real-time quantitative reverse transcription PCR (RT-qPCR). The chi-square analysis was used to analyze the correlation between MHECNR and miR-532-3p. Kaplan-Meier curve and multivariate Cox regression analysis were conducted to assess the significance of MHENCR in clinic. The interaction of MHENCR and miR-532-3p was probed using Pearson analysis and dual-luciferase reporter assay. Cellular experiments were implemented to explore the effects of MHENCR/miR-532-3p on colorectal carcinoma cells. Compared with para-cancerous tissues, MHENCR expression was increased and miR-532-3p expression was decreased in tumor tissues. High expression of MHENCR exhibited shorter overall survival. Interfering of MHENCR suppressed cellular activities while the silence of miR-532-3p diminished the decreased cellular behaviors in colorectal carcinoma cells. Interfering with MHENCR expression represses colorectal carcinoma cell proliferation, migration, and invasion by regulating miR-532-3p. MHENCR may act as a novel prognostic marker in colorectal carcinoma and MHENCR/miR-532-3p may serve as a potential target for treating colorectal carcinoma.

Deep learning models for image and data processes of intracellular calcium ions.

Intracellular calcium ion (Ca2+) in cytoplasm as an intracellular second messenger is involved in almost all important cellular activities of organisms. Generally its concentration ([Ca2+]i) is tested by live imaging followed image and data processes, in which much tedious and subjective manual work is involved. Here we show a computational approach of Deep Calcium following the principles of deep learning to predict the cytoplasmic Ca2+ ranges and calcium peaks in calcium curve of objective cells. To validate Deep Calcium, chondrocytes, bone marrow stromal cells (BMSCs) and osteoblastic like cells (MC3T3-E1) from both the tissue and cell samples as well as from spontaneous and mechanical stimulated calcium response patterns are used. The good performance comparing with other relative machine learning models, as well as consistency biological results with human experts are demonstrated. Deep Calcium provides references for other image and data processes of intracellular range determination and curve peak identification.

MicroRNA-92b acts as an oncogene by targeting PTEN/AKT in NSCLC.

MicroRNAs can act as tumour suppressors or oncogenes by regulating cellular differentiation, proliferation and apoptosis, and the dysregulation of miRNA is involved in the occurrence and development of NSCLC. Here, we provided evidence that miR-92b as an oncogene in NSCLC by targeting PTEN/AKT. We found that miR-92b was up-regulated in human NSCLC tissues and cell lines. MiR-92b knockdown suppressed the NSCLC cells proliferation and migration in both in vivo and in vitro models. Conversely, miR-92b overexpression induced an aggressive phenotype. Moreover, miR-92b-mediated regulation of NSCLC cell proliferation and migration depended on binding to PTEN mRNA, which then led to the degradation of PTEN and activation of the downstream AKT signalling pathway. Overall, this study revealed the oncogenic roles of miR-92b in NSCLC by targeting PTEN/AKT, and provided novel insights for future treatments of NSCLC patients. SIGNIFICANCE OF THE STUDY: MiR-92b was up-regulated in human NSCLC tissues and cell lines. Our study demonstrated that miR-92b as an oncogene in NSCLC by targeting PTEN/AKT in both in vivo and in vitro models and provided novel insights for future treatments of NSCLC patients.

Optimization and standardization of transient expression assays for gene functional analyses in strawberry fruits.

Strawberry is increasingly used as a model plant for research on fruit growth and development. The transient gene manipulation (TGM) technique is widely used to determine the function of plant genes, including those in strawberry fruits. However, its reliable application for the precise identification of gene function has been difficult owing to the lack of conditional optimization. In this study, we found that successful transient gene manipulation requires optimization, with the vector type, temperature, and fruit developmental stage being three major factors determining success. Notably, we found that transient gene manipulation was feasible only from the large green fruit stage onwards, making it especially suitable for identifying genes involved in strawberry fruit ripening. Furthermore, we established a method called percentage difference of phenotype (PDP), in which the functional effect of a gene could be precisely and efficiently identified in strawberry fruits. This method can be used to estimate the functional effect of a gene as a value from 0 to 100%, such that different genes can be quantitatively compared for their relative abilities to regulate fruit ripening. This study provides a useful tool for accelerating research on the molecular basis of strawberry fruit ripening.

The Lineage Specification of Mesenchymal Stem Cells Is Directed by the Rate of Fluid Shear Stress.

The effective regulation of fluid shear stress (FSS) on the lineage specification of mesenchymal stem cells (MSCs) remains to be addressed. We hypothesized that when MSCs are recruited to musculoskeletal system following stimulation, their differentiation into osteogenic or chondrogenic cells is directed by the rate of FSS (ΔSS) through modulation of the mechanosensitive, cation-selective channels (MSCCs), intracellular calcium levels, and F-actin. To this end, MSCs were exposed to laminar FSS linearly increased from 0 to 10 dyn/cm(2) in 0, 2, or 20 min and maintained at 10 dyn/cm(2) for a total of 20 min (termed as ΔSS 0-0', 0-2', and 0-20', respectively, representing more physiological (0-0') and non-physiological (0-2' and 0-20') ΔSS treatments). Our results showed 0-0' facilitated MSC differentiation towards chondrogenic and not osteogenic phenotype, by promoting moderate intracellular calcium concentration ([Ca(2+) ]i ) increase from the calcium channels with the exception of MSCCs or intracellular calcium stores, and F-actin organization. In contrast, 0-2' promoted MSCs towards osteogenic and not chondrogenic phenotype, by inducing significant [Ca(2+) ]i increase mainly from the MSCCs, and F-actin assembly. However, 0-20' elicited the modest osteogenic and chondrogenic phenotypes, as it induced the lowest [Ca(2+) ]i increase mainly from MSCCs, and F-actin assembly. Our results suggest that compared to the more physiological ΔSS, the non-physiological ΔSS favors [Ca(2+) ]i influx from MSCCs. An appropriate non-physiological ΔSS (0-2') even elicits a large [Ca(2+) ]i influx from the MSCCs that reverses the lineage specification of MSCs, providing validation for the high mechanosensitivity of MSCs and guidance for training osteoporosis and osteoarthritis patients. J. Cell. Physiol. 231: 1752-1760, 2016. © 2015 Wiley Periodicals, Inc.

Fe3O4@ZnO core-shell nanocomposites for efficient and repetitive removal of low density lipoprotein in plasma and on blood vessel.

Low density lipoprotein (LDL)-apheresis therapy, which directly removes LDL from plasma by LDL-adsorbents in vitro is found to be clinically effective and safe to lower the LDL content in blood to prevent cardiovascular disease. Thus, developing excellent LDL adsorbents are becoming more and more attractive. Herein, functional Fe3O4@ZnO core-shell nanocomposites have been synthesized by a facile and eco-friendly two-step method. Not only do they possess high LDL adsorption (in PBS/plasma as well as on blood vessels) and favorable magnetic targeting ability but they can also be reused conveniently, which offer the Fe3O4@ZnO core-shell nanocomposites significant potential in the removal of LDL in vitro and in vivo.

Magnetic Fe3 O 4 nanoparticles grafted with single-chain antibody (scFv) and docetaxel loaded ß-cyclodextrin potential for ovarian cancer dual-targeting therapy.

In order to improve the therapeutic efficiency and reduce the side effects on nonpathological cells and tissues, targeting drug delivery systems have gained more and more attraction. Here, we report a novel dual-targeting drug delivery system for ovarian cancer therapy. The inner core was made of iron oxide (Fe3O4) nanoparticles, synthesized by co-precipitation method. It was further surface-functionalized with amine groups to link single-chain antibody (scFv) and ß-cyclodextrin (ß-CD). Docetaxel (TXT) was finally included in the grafted ß-CD. FTIR and XPS confirmed the reactions. SEM found that the diameters of these Fe3O4 nanoparticles before and after functionalization were around 40 nm. Magnetization test showed that these particles were superparamagnetic. The in vitro release of TXT was concentration-driven and sustained, depending on the renewal rate of release medium. The in vitro flow chamber experiment revealed its magnetic targeting property; modified ELISA and static binding experiments displayed its good affinity to Endoglin, indicating that our drug delivery system has the potential to be dual-targeted to ovarian cancer tissue by externally applied magnetic field and native active binding of grafted scFv to Endoglin, overexpressed by ovarian cancer tissue. MTT assays showed that the TXT released from this drug delivery system continuously inhibited the growth of Skov3 ovarian cancer cells in 72h, better than the control raw TXT. All these results demonstrated a promising dual-targeting drug delivery system with great potential for ovarian cancer therapy.

Inserted rest period resensitizes MC3T3-E1 cells to fluid shear stress in a time-dependent manner via F-actin-regulated mechanosensitive channel(s).

The underlying cellular mechanism of anabolic effect recovered by inserting rest is not fully understood. In this work, we studied the role of F-actin regulated mechanosensitive channel(s) re-activation in mechanosensitivity modulation in vitro. Results showed that steady fluid shear stress (sFSS) stimulation with 30-min rest period was more potential in increasing alkalinephosphatase (ALP) activity than 10 and 0-min rest periods, and insertion of 30 min, but not 0 or 10 min, recovered the [Ca(2+)]i transient and contribution of the mechanosensitive channel(s). During the rest period, F-actin experienced polymerization (0-10 min), followed by depolymerization (10-30 min); inhibition of F-actin polymerization/depolymerization significantly increased/decreased the [Ca(2+)]i transient, as well as the contribution of the mechanosensitive channel(s) in subsequent sFSS stimulation. Our results demonstrated that the long rest period between sFSS loadings recruited [Ca(2+)]i transient via F-actin depolymerization-induced reactivation of mechanosensitive channel(s), suggesting that F-actin-regulated cellular stiffness could account for the decreased anabolic response during continuous mechanical loading in bone cells.

A confocal technique applicable to studies of cellular pH-related signaling in plants.

pH may act as a crucial signal in both animal and plant cells. It is very difficult to monitor pH signals and this has largely hindered progress in the investigation of pH signaling, particularly systematic pH signaling. Here, we report the development of a confocal technique to monitor leaf apoplastic pH in intact plants, which is particularly suitable for the studies on root to shoot signaling. A variety of different pH indicators and plant species were tested. It was found that different pH indicators, for example, 2',7'-Bis-(2-carboxyethyl)-5-(and-6)-carboxyfluoresce (BCECF), SNARF-4F 5-(and-6)-carboxylic acid (SNARF) and DM-NERF (NERF), were of different properties, and to successfully monitor pH at a sub-cellular level, the comparability between the pH indicator and plant species must be involved according to their suitable pH range and loading characteristics. The loading characteristics of different pH indicators differ with different plant species, cell types and their developing stages. No matter what methods were adopted, BCECF and SNARF could not be loaded specifically in the leaf apoplast in sunflower, tomato, and Comelina communis L. In contrast, regardless of the methods adopted, NERF could be loaded efficiently and specifically in the leaf apoplast in C. communis, but not in other plants. In C. communis, the determination coefficient for in vitro and in situ calibration of NERF was very high, which was respectively 0.9951 and 0.9916, and therefore, the adoption of NERF together with C. communis could construct an ideal experimental system that is suitable for the investigation of pH systematic signaling. Ratio image analysis demonstrated that the leaf apoplastic pH was about 5.5 in non-stressed conditions, and water deficit could trigger an increase in pH by about half a pH unit, which is the first evidence to directly indicate that pH is able to act as a systematic signal under water deficit conditions.

Dynamic analysis of ABA accumulation in relation to the rate of ABA catabolism in maize tissues under water deficit.

The plant hormone abscisic acid (ABA) accumulates in plant tissues which experience water deficit (stress ABA). This study analysed its accumulation as a function of both synthesis and catabolism in maize tissues. By following the disappearance of the stress ABA when ABA synthesis was blocked by nordihydroguaiaretic acid (NDGA), the rate of the catabolism of stress ABA was determined. When compared with the catabolic rate of baseline (non-stress) ABA, stress ABA showed a catabolic rate >11 times higher. With such an elevated catabolic rate, it is proposed that the xanthophyll precursor pool may not be able to sustain the ABA accumulation, and such a proposition has been substantiated by further experiments where fluridone is used to limit the availability of upstream ABA precursors. When fluridone was used, stress ABA accumulation could only be sustained for a few hours, i.e. approximately 5 h for leaf and 1 h for root tissues. In detached roots, stress ABA accumulation could not be sustained even if fluridone was not used, suggesting that stress ABA accumulation in root systems requires the continuous import of ABA precursors from the shoots. Such an assumption was substantiated by the observation that defoliation or shading significantly reduced ABA accumulation in intact roots. The present study suggests that ABA catabolism is rapid enough to play an important role in the regulation of ABA accumulation.