Schematic illustration of (A) Acr-Bis polymerization catalyzed by HRP /H2O2/ACAC ternary initiation system and (B) the polymerization of Acr-Bis causes AIE phenomenon of BSA-Au NCs.

The application of nonconventional nonaromatic luminophores-based hydrogels (NLHs) in bioanalysis is severely limited due to their luminescence in concentrated solution or solid state. Herein, a nonconventional NLH (B-Au@PDMAA) with high signal-to-noise ratio and stability in dilute solutions is designed by enhancing nonaromatic luminophores intermolecular aggregation through bovine serum albumin-gold nanoclusters (B-Au) confined space interaction. Importantly, the established NLHs could be rapidly polymerized in situ on magnetic beads (MB) surface in dilute solutions, and the fluorescence intensity after polymerization is 5.63-fold higher than that of direct polymerization. Moreover, proposed B-Au@PDMAA@MB constructed homogeneous immunoassay platform is demonstrated to be highly sensitive for detecting hepatitis B surface antigen with a sensitivity of 3.39 ng mL-1, which is 28.6-fold lower than that of B-Au@PDMAA based immunoassay and comparable to the sensitivity of the chemiluminescence immunoassay. This work presents a new strategy for the synthesis of NLHs with high brightness and stability in dilute solutions, and opens new avenues for the construction of novel homogeneous immunoassay platforms. Bovine serum albumin-gold nanoclusters (B-Au) can be used to improve the sensitivity and stability of nonconventional nonaromatic luminophores hydrogels-based immunoassay by two aspects, namely, 1) etched B-Au enhanced the blue, yellow, and red fluorescence, and 2) confined space interaction originated from poly-N,N-dimethylacrylamide/BSA hydrogen bond interaction, BSA hydrophobic domain-limiting effect, and gold atom-based coordination interaction.image

Bovine serum albumin-stabilized Au nanoclusters (BSA-Au NCs) have emerged as promising contenders for imaging agents and highly sensitive fluorescence sensors due to their biocompatibility and strong photoluminescence. Optimizing the synthesis conditions of BSA-Au NCs is crucial for enhancing fluorescence imaging and other nanocluster applications. In this study, for the first time, we systematically investigated the effects of BSA concentration and Au3+ on both particle size and optical characteristics of BSA-Au NCs. When the two components achieved a suitable concentration ratio, it was beneficial to form BSA-Au NCs with a high quantum yield (QY = 74.30%) and good fluorescence stability. In contrast, an inappropriate concentration ratio would lead to the formation of gold nanoparticles (Au NPs), and their internal filtration effect (IFE) would attenuate the fluorescence emission of BSA-Au NCs. The BSA-Au NCs were then employed as efficient fluorescence sensors for detecting Hg2+. Furthermore, the growth mechanism of BSA-Au NCs was elucidated by monitoring fluorescence changes during different incubation times. The BSA-Au NCs with a high quantum yield introduce a novel synthetic concept for sensitive fluorescent probes and expanding versatile applications of BSA-Au NCs in catalysis, chemical sensing and biomedicine.

Protein-stabilized gold nanoclusters (Prot-Au NCs) have been widely used in biosensing and cell imaging owing to their excellent optical properties and low biotoxicity. However, several Prot-Au NCs reported in the literature do not retain the biological role of the protein, which greatly limits their ability to directly detect biomarkers. This study demonstrated for the first time the successful synthesis of dual-function avidin-stabilized gold nanoclusters (Av–Au NCs) using a one-pot method. The resulting Av–Au NCs exhibited intense blue and red emissions under 374 nm excitation. Furthermore, the Av–Au NCs retained the native functionality of avidin to bind to biotin. When DNA strands modified with biotin at both ends (i.e., linker chains) were mixed with Av–Au NCs, large polymers were formed, indicating that Av–Au NCs could achieve fluorescence signal amplification by interacting with biotin. Taking advantage of the aforementioned properties, we constructed a novel enzyme-free fluorescent biosensor based on the Av–Au NCs-biotin system to detect DNA. The designed fluorescent biosensor could detect target DNA down to 0.043 nM, with a wide line range from 0.2 nM to 20 µM. Thus, these dual-functional Av–Au NCs were shown to be an excellent fluorescent material for biosensing.

Hepatitis B virus (HBV) infection remains a global public health problem, which is directly associated with a very high risk of evolving into liver cirrhosis and hepatocellular carcinoma (HCC). So far, enzyme-linked immunosorbent assay (ELISA) is still a main detection method for HBV infection in hospital, by which it enables to identify HBV infection in vitro through detection of circulating hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg). However, ELISA requires antibody-dependent and enzyme-labeling. What's worse, ELISA is silent to the type of occult HBV infection, which does not express HBsAg. Thus, it is of significance to establish a favorable alternative to detect HBV-related specific DNA rather than proteins, as well as to avoid antibody and enzyme-labeling assay for unambiguous screening of HBV infection. Herein, we have exploited a ternary nanocube (AgAuPt NCs)-based "off-on" blinking-type electrochemiluminescence (ECL) biosensor towards detection of HBV-related DNA for the first time, integrated with Pb2+-requiring DNAzyme recycling amplification. By this way, the detection limit down to 65 aM is achieved. The unique "off-on" blinking-type ECL biosensor not only overcomes the drawbacks of ELISA towards detection for HBV infection but also provides a potential for clinical application.

精准医疗时代背景下的新型分子诊断技术和方法为肝癌的个体化诊疗提供了新的契机,通过检测原发或转移性肝癌自发脱落或外源因素诱导脱落进入外周血液循环系统中的肿瘤细胞,即循环肿瘤细胞(CTCs),可为肝癌的早期筛查、辅助诊断、预后判断、疗效评估和复发转移等提供重要的指导.与传统检测方法相比,CTC检测具有无创、实时、方便、高效等显著特点,有望为肝癌患者开辟一条个性化的精准治疗之路.本文将对CTC在肝癌精准诊疗中的研究进展作一概述和展望,以期为临床提供重要的参考.

Thousands of long non-coding RNAs (lncRNAs) have been discovered in human genomes by gene chip, next-generation sequencing, and/or other methods in recent years, which represent a significant subset of the universal genes involved in a wide range of biological functions. An abnormal expression of lncRNAs is associated with the growth, invasion, and metastasis of various types of human cancers, including hepatocellular carcinoma (HCC), which is an aggressive, highly malignant, and invasive tumor, and a poor prognosis in China. With a more in-depth understanding of lncRNA research for HCC and the emergence of new molecular-targeted therapies, the diagnosis, treatment, and prognosis of HCC will be considerably improved. Therefore, this review is expected to provide recommendations and directions for future lncRNA research for HCC.

Hepatocellular carcinoma (HCC) is a life-threatening malignant tumor with high cancer mortality, therefore it is particularly important to sensitively detect the biomarkers of hepatocellular carcinoma for early diagnosis and treatment. In this work, a nonenzymatic and ultrasensitive electrochemiluminescence (ECL) cytosensor for HepG2 cells detection is established based on the PtNi nanocubes-catalyzed tyramine signal amplification. PtNi nanocubes (PtNi NCs) are firstly reported to catalyze tyramine-luminol (Tyr-Lum) with the aid of H2O2 for covalently binding to membrane proteins of HepG2 cells, realizing the introduction of massive Tyr-Lum ECL luminophores for signal amplification. Besides, PtNi NCs could also enhance the ECL signal of luminol-H2O2 system. On the basis of bifunctional PtNi NCs, the developed ECL cytosensor for HepG2 cells detection shows a wider linear range of 1x10 to 1x10(5) cells mL(-1) with detection limit of 3 cells mL(-1). This proposed enzyme-free-mediated tyramine signal amplification inspires a new strategy for applications in the early diagnosis of cancer cells.

Accumulated evidence revealed that numerous long noncoding RNAs (lncRNAs) have been found to be involved in the development and progression of hepatocellular carcinoma (HCC). LINC00628, a member of lncRNAs, has been reported to act as a tumor suppressor in gastric cancer and breast cancer. However, its potential role in HCC still remains unknown. Herein, we characterized the function of LINC00628 in HCC. Our investigation has revealed that LINC00628 were dramatically decreased in HCC tissues and cells, and inhibited the migration and invasion of HCC cells in vitro and in vivo. Moreover, LINC00628 exerted its tumor suppressive function by repressing the vascular endothelial growth factor A (VEGFA) promoter activity. A highly conserved region element in LINC00628 was identified by a cross-species comparative analysis, which is required for LINC00628 exerted its function. Dual-luciferase reporter assay showed that the conserved sequence mediated the interaction with a specific region of VEGFA promoter, resulting in a decrease of VEGFA expression. In conclusion, our results demonstrated that LINC00628 could function as a tumor suppressor in HCC via its conserved sequence elements interacting with a particular region of VEGFA promoter, suggesting that LINC00628 may serve as a novel promising target for diagnosis and therapy in HCC.

Background: Emerging studies demonstrate that long noncoding RNAs (lncRNAs) play crucial roles in hepato-carcinogenesis through various mechanisms. LncRNA CCAT2 was a newly discovered lncRNA and amplified in several cancers. However, the mechanisms involved in function of CCAT2 in hepatocellular carcinoma (HCC) remain to be explored. Methods: CCAT2 expressions in HCC tissues and cell lines were measured by RT-qPCR. MTS assay, colony formation assay, wound-healing assay and transwell assay were used to explore the biological functions of CCAT2 on HCC cells proliferation and metastasis. Experiments in vivo were carried out to confirm these effects. The underlying mechanisms were analyzed by western blot and dual-luciferase reporter assay. Results: In this study, we found that CCAT2 were significantly elevated in HCC tissues and cell lines, and it promoted HCC cells proliferation and metastasis both in vitro and in vivo. Additionally, we identified that NDRG1 was a downstream target of CCAT2. Meanwhile, depletion of CCAT2 inhibited cellular proliferation and metastasis behaviors induced by NDRG1- overexpression. Analysis of mechanism underlying these effects revealed that CCAT2 increased the expression of NDRG1 by enhancing its promoter activity. Furthermore, the active region between CCAT2 and NDRG1 promoter was confirmed by dual-luciferase reporter assay. Conclusions: All these observations demonstrate that CCAT2 acts as an oncogene by up-regulating NDRG1, which may have the potential to be used as a promising prognostic biomarker and therapeutic target for HCC.

Background Emerging evidences have indicated that long noncoding RNAs (lncRNAs) play essential roles in the development and progression of cancers. Dysregulation of lncRNA MIR31HG has recently been reported in several types of cancers, and researches on the function of MIR31HG in cancers suggested that MIR31HG could act as either oncogene or tumor suppressor. But the functional involvement of MIR31HG has not been studied in hepatocellular carcinoma (HCC). Methods In this study, MTS assays, colony formation assay, Wound-healing assay, Transwell assy, and tumor xenografts experiments were used to identify biological effects of MIR31HG on HCC cells HCC proliferation and metastasis in vitro and in vivo. Dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were performed to show the interactions of MIR31HG and miR-575. The bioinformatics methods were completed to find the target genes of miR-575. And Dual-luciferase reporter assay and Western blot analysis were further used to confirm the target gene of miR-575. Results We found that overexpression of MIR31HG obviously suppressed HCC proliferation and metastasis in vitro and in vivo, whereas knockdown of MIR31HG had the opposite effects. Besides, overexpression of MIR31HG significantly decreased the expression of microRNA-575 (miR-575), which plays an oncogenic role in HCC. Moreover, dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay revealed that MIR31HG exerted tumor-suppressive functions by binding directly to miR-575, and there was a reciprocal inhibition between MIR31HG and miR-575 in the same RNA-induced silencing complex (RISC). Furthermore, overexpression of MIR31HG enhanced the expression of suppression of tumorigenicity 7 like (ST7L), which was identified as a downstream target gene of miR-575. Thus, MIR31HG positively regulated ST7L expression through sponging miR-575, and acted as tumor suppressor in HCC. Conclusions Overall, our study illuminates the role of MIR31HG as a miRNA sponge in HCC, and sheds new light on lncRNA-directed diagnostics and therapeutics in HCC.

Long non-coding RNAs (lncRNAs) have emerged as critical regulators in a variety of diseases, including many tumors, such as hepatocellular carcinoma (HCC). However, the function and mechanisms responsible for these molecules in HCC are not thoroughly understood. In our previous study, we found that LINC00052 was acted as a tumor suppressor in HCC. In this study, we performed transcription microarray analysis to investigate the target gene of LINC00052, and found that knockdown of LINC00052 significantly increased the expression of SRY-related HMG-box gene 9 (SOX9), which plays an oncogenic role in HCC. Moreover, luciferase reporter assay revealed that LINC00052 promoted miR-101-3p expression by enhancing its promoter activity. In addition, online database analysis tools and luciferase assays showed that miR-101-3p could target SOX9. Quantitative real-time polymerase chain reaction (qRT-PCR) demonstrated that miR-101-3p was downregulated in HCC tissues and HCC cell lines. And we found a positive relationship between LINC00052 and miR-101-3p, and a negative relationship between miR-101-3p and SOX9 in HCC tissues. Besides, miR-101-3p was involved in LINC00052 inhibits HCC cells proliferation and metastasis. At the molecular level, LINC00052 downgulated SOX9 to inhibit HCC cells proliferation and metastasis by interacting with miR-101-3p. It might be a potential application for HCC therapy.

Herein, a novel and pragmatic electrochemiluminescence (ECL) biosensing method was developed for ultra sensitive and specific detection of Group B Streptococci (GBS) by combining self-enhanced luminol complex functionalized CuMn-CeO2 (CuMn-CeO2-PEI-luminol) with MNAzyme-mediated target-recycling amplification. First, the efficient self-enhanced PEI-luminol luminophore was prepared by combining PEI co-reactant with luminol in one molecular, which shortened electron transfer distance and enhanced ECL signal. And CuMn-CeO2 was applied to load a large number of PEI-luminol and strengthen luminous efficiency of luminol by the high catalytic activity toward H2O2 oxidation. Then, target-driven MNAzyme system was used to realize the circulation of GBS nucleic acid sequence, producing plentiful triggers to initiate the hybridization reaction on the surface of electrode. The developed enzyme-free ECL biosensor showed ultra-sensitivity for target DNA detection with detection limits of 68 aM (synthetic DNA) and 5 x 10(2) CFU mL(-1) (genomic DNA extracted from GBS strain). More importantly, this biosensor was successfully applied for detection of genomic DNA of GBS extracted from clinical vaginal/anal swabs as low as 320 copies. Thus, this proposed strategy might be an pragmatic ECL platform for ultrasensitive and specific detection of GBS in clinical vaginal/anal swabs.