Smartphone-based colorimetric sensor array using gold nanoparticles for rapid distinguishment of multiple pesticides in real samples.

A simple, sensitive method for pesticide distinguishment based on a colorimetric sensor array using diverse gold nanoparticles (AuNPs) at room temperature is presented in this study. Acetylcholinesterase (AChE) hydrolysis ability was influenced by different pesticides and produced different concentrations of thiocholine by hydrolyzing acetylthiocholine iodide (ATCh). Thiocholine could be easily linked to the AuNPs through an Au - S covalent bond, and AuNPs underwent aggregation, resulting in a visible color change due to alteration of surface plasmon resonance properties. Based on these results, we successfully distinguished eight pesticides (glyphosate, thiram, imidacloprid, tribenuron methyl, nicosulfuron, thifensulfuron methyl, dichlorprop, and fenoprop) utilizing five different AuNPs by colorimetric assay. The limit of detection (LOD) of this visual method for all pesticides was less than 1.5 × 10-7 M, which was more sensitive than the U.S. Environmental Protection Agency regulations specify (1.18 âˆ¼ 3.91 × 10-6 M). This method was further improved by combining a portable smartphone device with a color picking application using (color name AR) and RGB (red, green, blue) values. The method was successfully applied to pesticide residue distinguishment in real samples by linear discriminant analysis (LDA).

Circular RNA hsa_circ_0000437 may be used as a new indicator for the diagnosis and prognosis of hepatocellular carcinoma.

Circular RNAs (circRNAs) play an essential role in hepatocellular carcinoma (HCC); however, the precise role of circRNAs in the diagnosis and prognosis of HCC remains unclear. The circRNA circ_0000437 was identified in the microarray dataset GSE166678 and was detected in HCC and paired adjacent tissue and serum samples in both the HCC and control groups by reverse transcription quantitative PCR. The association between circ_0000437 expression and clinicopathological characteristics was investigated. Furthermore, the diagnostic and prognostic values of circ_0000437 were determined using receiver operating characteristic (ROC) and Kaplan-Meier curves. Circ_0000437 expression was markedly upregulated in the tumor group compared with the control group and was correlated with tumor node metastasis (TNM) classification, differentiation degree, tumor size, and Barcelona Clinic Liver Cancer (BCLC) stage (P< 0.05) in both the tumor tissues and serum. Furthermore, poor overall survival (OS) was correlated with high circ_0000437 expression, and the area under the ROC curve (AUC) of circ_0000437 for the diagnosis of HCC was 0.9281 in the serum. Our findings suggest that circ_0000437 may be used as a novel biomarker for the diagnosis and prognosis of patients with HCC.

Bioinformatics analysis of tumor-educated platelet microRNAs in patients with hepatocellular carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most prevalent malignancies that seriously threaten global health. The primary reason for its grim prognosis is the lack of sensitive tools for early diagnosis. The purpose of the present study was to apply bioinformatics analysis to explore tumor-educated platelet (TEP) microRNA (miRNA) expression and its potential diagnostic utility in HCC. METHODS: Twenty-five HCC patients and 25 healthy controls were included. RNA sequencing was utilized to screen miRNA alterations in platelets derived from HCC patients (n=5) and controls (n=5). Gene set enrichment analysis was performed to analyze the targeted mRNAs of differentially expressed miRNAs by using the Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) databases, aiming at main functions and pathways, respectively. We then verified the selected platelet miRNAs in another cohort by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) amplification. RESULTS: A total of 250 differentially expressed miRNAs were identified, among which 111 were down-regulated and 139 were up-regulated. The functional enrichment analysis of differentially expressed miRNAs suggested that their target genes were involved primarily in pathways related to HCC. Expression levels of miR-495-3p and miR-1293 were further validated by qRT-PCR, which yielded results consistent with the sequencing analysis. The area under the receiver operating characteristic (ROC) curve of miR-495-3p and miR-1293 as diagnostic tests for HCC were 0.76 and 0.78, respectively. CONCLUSION: TEP miRNAs such as miR-495-3p and miR-1293 were differentially expressed in HCC patients, and may be involved in the pathophysiology of HCC.

Aggregation-induced enhanced emission-type cruciform luminophore constructed by carbazole exhibiting mechanical force-induced luminescent enhancement and chromism.

One D-A type cruciform luminophore MDCS-BC based on carbazole has been prepared. We observed that this compound exhibits unique intramolecular charge-transfer (ICT) and typical aggregation-induced enhanced emission (AIEE) properties with the solid-state luminescence efficiency of 0.184. Moreover, this luminophore shows a significant stimuli-induced emission enhancement and chromism effect with good reversibility. Upon grinding, the fluorescence color of the as-prepared samples can change from blue (454 nm) to green (504 nm). What is unexpected is that the fluorescence efficiency of the initial powder is dramatically increased from 0.184 to 0.424 upon grinding. The results of PXRD, DSC and spectral properties studies show that the mechanical force-induced luminescence enhancement and chromism behavior of MDCS-BC originates from the transition between crystal and amorphous morphology, and the large red-shift and the emission enhancement inducing by grinding may be attributed to the planarization of the molecular conformation and subsequent planar ICT process.

Percutaneous Microwave Ablation in the Spleen for Treatment of Hypersplenism in Cirrhosis Patients.

AIM: The aim of this study was to estimate the feasibility and therapeutic effectiveness of percutaneous microwave ablation in the treatment of hypersplenism in cirrhosis. METHODS: Forty-one cirrhosis patients with hypersplenism were treated with ultrasonography-guided percutaneous microwave ablation between February 2007 and August 2011. Peripheral blood cell counts, portal vein diameter, splenic vein diameter, and blood flow of splenic vein were evaluated before and after the operation, and complications of the treatment were also investigated. All patients were followed up for 24 months. RESULTS: The levels of platelets and white blood cells were increased, while the splenic vein diameter narrowed gradually after the therapy and 24 months later. Moreover, patients received percutaneous microwave ablation had much lower splenic venous flow velocity. The portal vein diameter did not change significantly 6 months after the treatment, although it narrowed gradually within 3 months after the treatment. Furthermore, no complications such as uncontrollable bleeding, splenic abscess, spleen rupture, and damage in surrounding organ happened after the therapy. CONCLUSIONS: Graded percutaneous microwave ablation, as a minimally invasive therapy, could damage the spleen, increase the levels of platelets and white blood cells, and reduce portal hypertension effectively without serious complications. Percutaneous microwave ablation is an effective, safe, and feasible method for cirrhosis patients with hypersplenism.

Hepatitis B virus X protein promotes P3 transcript expression of the insulin-like growth factor 2 gene via inducing hypomethylation of P3 promoter in hepatocellular carcinoma.

BACKGROUND & AIMS: Hepatitis B virus (HBV) X protein (HBx) contributes to hepatocarcinogenesis. The overexpression of transcripts from P3 and P4 promoters of the insulin-like growth factor 2 (IGF2) gene is observed in hepatocellular carcinoma (HCC). Here, we aimed to explore the involvement of HBx in P3-driven mRNA overexpression and underlying epigenetic mechanism. METHODS: P3 mRNA, P3 methylation status, HBx mRNA and HBx protein were analysed in human HCC samples with and without HBV infection using quantitative RT-PCR, bisulphite sequencing and Western blotting. The effects of HBx on P3 mRNA expression, and P3 transcriptional activity and methylation were further evaluated in HCC cell lines. RESULTS: P3 mRNA level was higher and P3 methylation level was lower in HBV-positive HCC specimens compared with those of HBV-negative HCC specimens. P3 transcript abundance was positively correlated with HBx expression and negatively correlated with P3 methylation in HCC specimens. The stable expression of HBx upregulated P3 mRNA expression and reduced P3 methylation level in HepG2-HBx cells. The transient expression of HBx stimulated P3 promoter activity and decreased P3 methylation level of P3 promoter-luciferase construct in a dose-dependent manner in HepG2 and Huh-7 cells. Furthermore, HBx mRNA expression was found to be independent predictive factors for both shorter disease-free survival time and shorter overall survival time of HCC patients. CONCLUSION: HBx may promote IGF2-P3 transcript expression by inducing hypomethylation of P3 promoter and may be associated with an inferior clinical outcome of HBV-related HCC patients. This study provides useful information for understanding the mechanism of HBx-mediated HCC.

[Hepatitis B virus X protein promotes insulin-like growth factor II gene expression by inducing hypomethylation of the P3 promoter in hepatocellular carcinoma].

OBJECTIVE: To explore the involvement of hepatitis B X protein (HBx) in promoter 3 (P3)-driven mRNA overexpression of the insulin-like growth factor II gene (IGF-II) and investigate the underlying epigenetic mechanism. METHODS: Levels of P3 and HBx mRNA and status of P3 methylation were analyzed in human hepatocellular carcinoma (HCC) samples, with and without hepatitis B virus (HBV) infection, using quantitative reverse transcription-PCR and bisulfite sequencing. In addition, the levels of P3 mRNA and P3 methylation were examined in HepG2 cells stably overexpressing HBx (HepG2-HBx). Finally, P3 promoter-luciferase constructs were cotransfected into HepG2 cells along with an HBx-expressing plasmid, and the effects of HBx on transcriptional activity and methylation of P3 were analyzed. Statistical analyses of the data were conducted by chi square test, Fisher's exact test, Student's t-test, Marn-Whitney U test, and Pearson's correlation coefficient test. RESULTS: The HBV-positive HCC specimens had significantly higher levels of P3 mRNA than the HBV-negative HCC specimens (-9.59 ± 3.22 vs. -12.97 ± 3.08 delta CT; P=0.006) but significantly lower levels of P3 methylation (mean values for the 17 CpG sites (36.9% ± 15.5% vs. 52.1% ± 19.1%; P=0.025). The P3 transcript abundance was positively correlated with the level of HBx expression and negatively correlated with the level of P3 methylation. The epigenetic results from experiments with the HepG2-HBx cells were similar. Transfection of HBx significantly decreased P3 methylation level and increased its activity. CONCLUSION: HBx expression may promote IGF-II expression by inducing hypomethylation of its P3 promoter in hepatocellular carcinoma.

[The construction of IGF-II P4 promoter-driven tk expression vector].

OBJECTIVE: To investigate the selective killing effect of herpes simplex virus-thymidine kinase/ganciclovir (HSV-tk/GCV) suicide gene system controlled by human IGF-II P4 promoter on HCC cells in vitro. METHODS: Recombinant shuttle plasmid vectors driven by IGF-II P4 promoter and driven by CMV promoter were constructed by techniques of gene recombination. The recombinant shuttle plasmids were then transfected into HepG2 and HeLa cells by techniques of lipidosome transfection. EGFP expression was detected by fluoroscopy. Tk and EGFP mRNA expression were detected by RT-PCR. The selective killing effect after GCV application was determined with MTT method. Statistical analysis was performed with ANOVA analysis. RESULTS: Identification of pDC316-tkEGFP-P4 by enzyme digestion and sequencing analysis showed that the construction of the recombinant shuttle plasmid was correct. It was found that green fluorescence protein could only be seen in HepG2 cells, not in HeLa cells. The results of RT-PCR showed only two bands could be seen in the samples of pDC316-tkEGFP-P4 transfected HepG2 cells. The growth of HepG2 cells transfected with pDC316-tkEGFP-CMV and pDC316-tkEGFP-P4 were inhibited remarkably, the growth inhibition rates were 6.95% +/- 0.67%, 24.99% +/- 1.53%, 49.68% +/- 1.68%, 71.85% +/- 3.28% and 4.83% +/- 0.35% vs 17.34% +/- 1.15%, 30.17% +/- 1.30%, 40.39% +/- 0.82% (F = 24.055, P < 0.05), respectively. The growth of HeLa cells transfected with pDC316-tkEGFP-CMV were also inhibited, the growth inhibition rates were 6.36% +/- 0.83%, 23.95% +/- 1.72%, 45.13% +/- 1.64% and 69.38% +/- 3.17%, respectively. The growth of HeLa cells transfected with pDC316-tkEGFP-P4 was not inhibited. The growth inhibition rates were 0.91 +/- 0.04, 1.18 +/- 1.32, 1.19 +/- 0.10 and 1.32 +/- 0.05 (F = 26.469, P < 0.01) , respectively. CONCLUSION: The shuttle plasmid vector carrying the tkEGFP fusion protein gene driven by IGF-II P4 promoter has been constructed successfully and its specific expression in HepG2 cells provided the basis for targeted gene therapy for HCC.

[Construction of a plasmid vector of fused protein genes driven by human insulin-like growth factor II P3 promoter].

OBJECTIVE: To construct a shuttle plasmid vector of fused herpes simplex virus thymidine kinase (HSV-tk) gene and enhanced green fluorescent protein (EGFP) gene driven by human insulin-like growth factor II (IGF-II) P3 promoter, and investigate the special killing effect of the HSV-tk/ganciclovir (GCV) system on hepatocellular carcinoma (HCC) cells. METHODS: An adenovirus shuttle plasmid, pDC316-tkEGFP-CMV containing fused genes tkEGFP and an adenovirus shuttle plasmid pDC316-tkEGFP-P3 driven by IGF-II P3 promoter were constructed by techniques of gene recombination and screening, and identified by restriction digestion and sequencing analysis. Human hepatocellular carcinoma cells HepG2 and human cervical carcinoma cells HeLa were cultured and transfected with these 2 recombinant shuttle plasmids. RT-PCR was used to detect the mRNA expression of EGFP and HSV/tk. GCV of the final concentrations of 0, 1, 10, and 100 microg/ml respectively was added into the culture fluid of the HepG2 cells transfected with pDC316-tkEGFP-CMV or pDC316-tkEGFP-P3, and MTT method was used to detect the cell inhibition rate. RESULTS: Digestion and sequencing analysis showed that the recombinant plasmid pDC316-tkEGFP-P3 accorded with the design. Fluorescent microscopy showed that EGFP was expressed only in the HepG2 cells, but not in the HeLa cells. RT-PCR showed that mRNA expression of EGFP and HSV/tk could be seen in both HepG2 and HeLa cells transfected with pDC316-tkEGFP-CMV or pDC316-tkEGFP-P3, however, only in the pDC316-tkEGFP-P3 transfected HepG2 cells, but not in the HeLa cells transfected with pDC316-tkEGFP-P3. MTT assay showed that GCV dose-dependently inhibited the 2 cancer cells, the inhibition rates of GCV of the final concentrations of 1, 10, and 100 microg/ml were 24.1% +/- 1.9%, 45.1% +/- 1.7%, and 69.4% +/- 3.6% in the HepG2 cells, and 25.1% +/- 1.6%, 49.3% +/- 1.1%, and 72.2% +/- 2.9% in the HeLa cells. However, the inhibition rates of the pDC316-tkEGFP-P3-transfected HepG2 cells by GCV of the final concentrations of 1, 10, and 100 microg/ml wee 19.8% +/- 1.3%, 36.2% +/- 2.0% and 48.7% +/- 1.9% respectively, all significantly lower than those of the pDC316-tkEGFP-CMV-transfected HepG2 cells (all P < 0.01), and no significant cell inhibition was found in the HeLa cells transfected with pDC316-tkEGFP-CMV. CONCLUSION: A shuttle plasmid vector containing the tkEGFP fusion protein gene driven by IGF-II P3 promoter has been constructed successfully and its specific expression in HepG2 cells provides a sound basis for targeted gene therapy for HCC.