A novel and sensitive ratiometric fluorescent quantum dot-based biosensor for alkaline phosphatase detection in biological samples via the inner-filter effect.

Alkaline phosphatase (ALP) is an important biomarker whose abnormal level in activity is associated with hepatobiliary, skeletal, and renal diseases as well as cancer. Herein, we synthesized ZnSe@ZnS quantum dots (ZnSe@ZnS QDs) and Mn-doped ZnS quantum dots (Mn:ZnS QDs) as fluorophores to establish the ratiometric fluorescent assay for ALP activity detection in biological samples. p-Nitrophenyl phosphate (PNPP) was used as a substrate for ALP, and the overlaps between absorption spectra of PNPP and excitation spectra of QDs resulted in sharp fluorescence quenching. Under the catalysis of ALP, PNPP was hydrolyzed into p-nitrophenol (PNP), which caused a red shift of absorption band of PNPP and fluorescence recovery of Mn:ZnS QDs (585 nm). However, the overlaps between absorption spectra of PNP and emission spectra of ZnSe@ZnS QDs led a further quenching of ZnSe@ZnS QDs (405 nm). Therefore, the ratiometric fluorescent signals (F 585/F 405) were associated with activity of ALP based on bidirectional responses of QDs to the concentration of PNPP. Under the optimum conditions, the method exhibited a good linear relationship from 4 to 96 U per L (R 2 = 0.9969) with the detection limit of 0.57 U per L. Moreover, the method was successfully applied for detecting the ALP activity in a complex biological matrix (human serum and HepG2 cells) with impressive specificity. In particular, the complicated chemical modifications of QDs and pretreatments of biological samples were not required in the whole detection procedures. Therefore, it not only provided a sensitive, specific and simple approach to clinical ALP activity detection, but it also provided support for early diagnosis of diseases.

Learning Single/Multi-Attribute of Object with Symmetry and Group.

Attributes and objects can compose diverse compositions. To model the compositional nature of these concepts, it is a good choice to learn them as transformations, e.g., coupling and decoupling. However, complex transformations need to satisfy specific principles to guarantee rationality. Here, we first propose a previously ignored principle of attribute-object transformation: Symmetry. For example, coupling peeled-apple with attribute peeled should result in peeled-apple, and decoupling peeled from apple should still output apple. Incorporating the symmetry, we propose a transformation framework inspired by group theory, i.e., SymNet. It consists of two modules: Coupling Network and Decoupling Network. We adopt deep neural networks to implement SymNet and train it in an end-to-end paradigm with the group axioms and symmetry as objectives. Then, we propose a Relative Moving Distance (RMD) based method to utilize the attribute change instead of the attribute pattern itself to classify attributes. Besides the compositions of single-attribute and object, our RMD is also suitable for complex compositions of multiple attributes and objects when incorporating attribute correlations. SymNet can be utilized for attribute learning, compositional zero-shot learning and outperforms the state-of-the-art on four widely-used benchmarks. Code is at https://github.com/DirtyHarryLYL/SymNet.

Integrated bioinformatic analysis and experiment confirmation of the antagonistic effect and molecular mechanism of ginsenoside Rh2 in metastatic osteosarcoma.

This study aimed to compare the gene expression variation of clinical primary osteosarcoma (OS) and metastatic OS, identify expression profiles and signal pathways related to disease classification, and systematically evaluate the potential anticancer effect and molecular mechanism of ginsenoside Rh2 on OS. A raw dataset (GSE14359), which excluded GSM359137 and GSM359138, was downloaded from the Gene Expression Omnibus. Differentially expressed genes (DEGs) and principal component analysis (PCA) were obtained with limma. Pathways enrichment analysis was understood by GSEA app. Rh2-associated targets were harvested and mapped through PharmMapper and Cytoscape 3.4.0. The toxicity of Rh2 was determined using crystal staining and MTT assay on 143B and MG63 cell lines. The relative protein expression was confirmed through Western blot analysis. The mitochondrial membrane potential (△Ψm) was evaluated by JC-1 fluorescence staining. The cell mobility was measured via wound healing and transwell assays. A total of 752 genes were upregulated, while 161 genes were downregulated. GSEA and PCA displayed significant function enrichment and classification. Through PharmMapper and Cytoscape 3.4.0, Rh2 was found to target the mitogen activated protein kinase (MAPK) and PI3K signaling pathways, which are the key pathways in the metastasis of OS. Furthermore, Rh2 induced a concentration-dependent decrease in cell viability and early apoptosis associated with ΔΨm decline, while a non-lethal dose of Rh2 weakened the metastatic capability. Moreover, systematic evaluation showed that promoting the MAPK signaling pathway and inhibiting PI3K/Akt/mTOR were correlated with the anticancer effects of Rh2 on metastatic OS. In conclusion, transcriptome-derived approaches may be beneficial in diagnosing early metastases, and Rh2, a multi-targeting agent, shows promising application potential in suppressing metastatic OS in an MAPK- and PI3K/Akt/mTOR-dependent manner.

The microRNA­708­5p/ZEB1/EMT axis mediates the metastatic potential of osteosarcoma.

MicroRNA­708­5p (miR­708­5p) and epithelial­â€‹to­mesenchymal transition (EMT) have been widely identified to contribute to the pathogenesis and progression of multiple cancers. However, the connection between miR­708­5p and EMT has not been sufficiently clarified. Therefore, our research aimed to investigate the impact of miR­708­5p on EMT and the metastasis of osteosarcoma (OS). We first analyzed the differentially expressed microRNAs (DEmiRNAs) from the GSE70367 dataset. We found that the expression of miR­708­5p was lower in OS cells. Overexpression of miR­708­5p was able to impair the migration and invasion of OS cells. Moreover, miR­708­5p inhibited EMT of OS cells MG63 and SaOS­2, wherein E­cadherin was increased, and N­cadherin, vimentin, and Snail were decreased. Semaphorin 4C (SEMA4C), mitogen­activated protein kinase kinase kinase 3 (MAP3K3), and zinc finger E­box­binding homeobox 1 (ZEB1) were predicted as target genes of miR­708­5p by bioinformatics method. Only ZEB1, one of the EMT­inducing transcription factors, was validated as the direct target gene of miR­708­5p in OS cells through dual­luciferase reporter assay and Western blot analysis. Knockdown of ZEB1 was found to inhibit the metastasis of MG63 and SaOS­2 cells, whereas ZEB1 over-expression promoted their metastasis. In summary, miR­708­5p impaired the metastasis and EMT of OS, which was found to be mediated by inhibition of ZEB1.

Ginsenoside Rg3 Inhibits the Growth of Osteosarcoma and Attenuates Metastasis through the Wnt/ß-Catenin and EMT Signaling Pathway.

Osteosarcoma (OS) is the most common primary malignant bone cancer. An increasing number of studies have demonstrated that ginsenoside Rg3 (Rg3), which is extracted from the roots of the traditional Chinese herb Panax ginseng, plays a tumor suppression role in various malignant tumors. In the present study, we aimed at investigating the role of Rg3 in the proliferation, migration, and invasion of OS and at exploring the underlying mechanisms. Cell viability and proliferation were observed by MTT assay and crystal violet staining. The migration and invasion of cells were measured by wound-healing assay and Transwell method. Signaling pathway screening was investigated using luciferase reporter gene assay. qRT-PCR and western blot were performed to measure the expression of molecules involved in cell epithelial-mesenchymal transition (EMT), and Wnt/ß-catenin pathway. Results suggested that Rg3 could not only inhibit proliferation but also hamper the migration and invasion of OS. qRT-PCR and western blot demonstrated that a reduced level of MMP2/MMP7/MMP9 was induced after Rg3 treatment. In addition, the expression levels of proteins related to EMT and the Wnt/ß-catenin pathway were downregulated. In summary, our data revealed that Rg3 could inhibit the proliferation, migration, and invasion of OS cells. This effect of Rg3 might be mediated by downregulating MMP2, MMP7, and MMP9 expression and suppressing EMT as well as the Wnt/ß-catenin pathway. Thus, Rg3 might be a potential agent for the treatment of OS.