Programmable Microparticle Array for In Situ Modification and Multiple miRNA Detection.

Simultaneous detection of multiple miRNAs of one disease can greatly reduce misdiagnosis and improve the detection rate, which is helpful for early cancer diagnosis. Here, a programmable microparticle-array-based acoustic microchip for in situ simultaneous multiple miRNAs detection is developed. On this microchip, the multiple probes-labeled microparticle array can be procedurally arranged in a microfluidic reaction chamber when four orthogonally piezoelectric transducers are applied. The probes-labeled microparticle array offers a platform for full molecular contact under dynamic ultrasonic streaming, and the array supplies a multipoint data correction to reduce the false positive of the detection results for more precisely visible fluorescence multiple target miRNAs sensing. We employed miRNA-21, miRNA-210, and miRNA-155 as specific biomarkers of pancreatic cancer and successfully finished the multiple miRNAs simultaneous detection in the microchip with a detection limit of 139.1, 179.9, and 111.4 pM, respectively. Such a device is programmable by adjusting the imputing frequency and voltage, and target biomarkers can be easily collected when the ultrasound force is released for further analysis, which shows great potential in multiple miRNAs enrichment and simultaneous detection for cancer clinical diagnosis.

Biomineralization-inspired magnetic nanoflowers for sensitive miRNA detection based on exonuclease-assisted target recycling amplification.

Biomineralization-inspired magnetic hybrid nanoflowers were prepared facilely, and capture probes were easily immobilized on the obtained nanoflowers without tedious processing. Based on the magnetic hybrid nanoflowers and exonuclease-assisted target recycling amplification, a fluorescence miRNA sensor was fabricated. The presence of target miRNA leads to the formation of the double-strand structure, which would then be selectively digested by the exonuclease and increase fluorescence intensity. The target miRNA can be released for recycling and signal amplification. Under optimized reaction conditions, the hybrid nanoflower-based miRNA sensor had a broad detection range from 0.001 nM to 100 nM and a limit of detection of 0.23 pM (S/N = 3). The sensitive detection of miRNA in serum was also achieved with recoveries from 94.3% to 116.1%. This work provides a new insight into the fabrication of bioconjugated materials and shows great potential in miRNA sensing.

Patterned Liquid-Infused Nanocoating Integrating a Sensitive Bacterial Sensing Ability to an Antibacterial Surface.

The slippery liquid-infused surfaces show a great antibacterial property. However, most liquid-infused surfaces cannot detect whether or not the unknown aqueous samples contain microorganisms. Therefore, it is highly necessary but a challenge to integrate bacterial sensing capability into antibacterial surface. In this work, we prepared a slippery patterned liquid-infused nanocoating on the glass substrate for integrating bacterial sensing capability into the bacterial repellence surface. Dendritic mesoporous silica nanoparticles (DMSNs) with a suitable particle size of ca. 128 nm were employed as a building block to fabricate the multifunctional nanocoating with a superhydrophilic microwell and hydrophobic periphery by a dip-coating strategy, hydrophobic treatment, photomask-mediated plasma etching, and liquid infusion. Dendritic porous silica nanoparticles (DPSNs) with a larger particle size of ca. 260 nm were uniformly loaded with Au nanoparticles (NPs), providing large surface area for the modification of Raman reporter (4-mercaptobenzoic acid (4-MBA)) and aptamer. Thus, as a Raman tag, the formed DPSNs-Au-MBA-aptamer could achieve sensitive surface-enhanced Raman spectroscopy (SERS) detection of target bacteria. Combined with the Raman tag, the patterned liquid-infused nanocoating not only completely repelled bacteria on the hydrophobic area but also enabled sensitive SERS detection of Staphylococcus aureus in a very low sample volume (1 µL) with a low detection limit of 2.6 colony formation units (CFU)/mL on the antibody-modified superhydrophilic microwell. This research provided a novel and reliable strategy to construct a multifunctional nanocoating with microbial repellence and sensing capabilities.

Pathogen manipulation of chloroplast function triggers a light-dependent immune recognition.

In plants and animals, nucleotide-binding leucine-rich repeat (NLR) proteins are intracellular immune sensors that recognize and eliminate a wide range of invading pathogens. NLR-mediated immunity is known to be modulated by environmental factors. However, how pathogen recognition by NLRs is influenced by environmental factors such as light remains unclear. Here, we show that the agronomically important NLR Rpi-vnt1.1 requires light to confer disease resistance against races of the Irish potato famine pathogen Phytophthora infestans that secrete the effector protein AVRvnt1. The activation of Rpi-vnt1.1 requires a nuclear-encoded chloroplast protein, glycerate 3-kinase (GLYK), implicated in energy production. The pathogen effector AVRvnt1 binds the full-length chloroplast-targeted GLYK isoform leading to activation of Rpi-vnt1.1. In the dark, Rpi-vnt1.1-mediated resistance is compromised because plants produce a shorter GLYK-lacking the intact chloroplast transit peptide-that is not bound by AVRvnt1. The transition between full-length and shorter plant GLYK transcripts is controlled by a light-dependent alternative promoter selection mechanism. In plants that lack Rpi-vnt1.1, the presence of AVRvnt1 reduces GLYK accumulation in chloroplasts counteracting GLYK contribution to basal immunity. Our findings revealed that pathogen manipulation of chloroplast functions has resulted in a light-dependent immune response.

The effect of arbuscular mycorrhizal fungi and biochar on the growth and Cd/Pb accumulation in Zea mays.

A greenhouse pot experiment was conducted to assess the effects of biochar (BC) and arbuscular mycorrhizal fungus (AMF)-Funneliformis mosseae (Fm), Glomus versiforme (Gv) and Rhizophagus intraradices (Ri) on the plant growth and Cd/Pb accumulation by corn grown in the soils artificially contaminated with 5 mg Cd and 300 mg Pb kg-1 soil. The single AMF inoculation and combined usage of AMF and BC evidently improved the P contents of maize. Furthermore, the combined use of AMF and BC produced pronounced positive effect on corn growth, and the shoot biomass in Gv + BC group was 9.85-fold higher than that of the control. Meanwhile, the single BC addition and combined utilization of AMF and BC significantly reduced Cd and Pb concentrations in maize, and the greater reduces were found in the combined utilization, and the lowest Cd concentration of shoot was appeared in Gv + BC group. The single BC addition and combined application of AMF and BC significantly increased soil pH, and reduced soil diethylenetriaminepentaacetic acid (DTPA)-extractable Cd/Pb. This study demonstrated a synergistic effect between AMF (Gv, Fm, Ri) and BC on improving maize growth and decreasing Cd/Pb accumulation in maize, and the combined use of Gv and BC brought the most pronounced effect, which could provide a feasible strategy for safe production of maize from Cd/Pb-polluted soils.

Combined application of arbuscular mycorrhizal fungi and steel slag improves plant growth and reduces Cd, Pb accumulation in Zea mays.

Little attention has been paid to the combined use of arbuscular mycorrhizal fungus (AMF) and steel slag (SS) for ameliorating heavy metal polluted soils. A greenhouse pot experiment was conducted to study the effects of SS and AMF-Funneliformis mosseae (Fm), Glomus versiforme (Gv) and Rhizophagus intraradices (Ri) on plant growth and Cd, Pb uptake by maize grown in soils added with 5 mg Cd kg-1 and 300 mg Pb kg-1 soil. The combined usage of AMF and SS (AMF + SS) promoted maize growth, and Gv + SS had the most obvious effect. Meanwhile, single SS addition and AMF + SS decreased Cd, Pb concentrations in maize, and the greater reductions were found in combined utilization, and the lowest Cd, Pb concentrations of maize appeared in Gv + SS. Single SS amendment and AMF + SS enhanced soil pH and decreased soil diethylenetriaminepentaacetic acid (DTPA)-extractable Cd, Pb concentrations. Furthermore, alone and combined usage of AMF and SS increased contents of soil total glomalin. Our research indicated a synergistic effect between AMF and SS on enhancing plant growth and reducing Cd, Pb accumulation in maize, and Gv + SS exerted the most pronounced effect. This work suggests that AMF inoculation in combination with SS addition may be a potential method for not only phytostabilization of Pb-Cd-contaminated soil but maize safety production.

Arbuscular mycorrhizal fungi alleviate Cd phytotoxicity by altering Cd subcellular distribution and chemical forms in Zea mays.

Arbuscular mycorrhizal fungus (AMF) can relieve Cd phytotoxicity and improve plant growth, but the mechanisms involved in this process have still been not completely known. In the present work, a pot experiment was conducted to examine productions of glutathione (GSH) and phytochelatins (PCs), and absorption, chemical forms and subcellular distribution of Cd in maize (Zea mays) inoculated with or without AMF (Rhizophagus intraradices (Ri) and Glomus versiforme (Gv)) in Cd-amended soils (0, 1 and 5 mg Cd kg-1 soil). In general, both Ri and Gv inoculation dramatically enhanced biomass production and reduced Cd concentrations in shoots and roots of maize when compared to the non-mycorrhizal treatment. Moreover, both Ri and Gv symbiosis obviously increased contents of GSH and PCs, both in shoots and roots. Subcellular distribution of Cd in maize indicated that most of Cd (more than 90%) was accumulated in cell wall and soluble fraction. In addition, Cd proportions in soluble fractions in shoots of maize inoculated with Gv or Ri were considerably increased, but reduced in cell wall fractions compared to non-mycorrhizal maize, indicating that mycorrhizal symbiosis promoted Cd transfer to vacuoles. Furthermore, proportions of Cd in inorganic and water-soluble forms were declined, but elevated in pectates and proteins-integrated forms in mycorrhizal maize, which suggested that Gv and Ri could convert Cd into inactive forms. These observations could provide a further understanding of potential Cd detoxification mechanism in maize inoculated with AMF.

An oomycete plant pathogen reprograms host pre-mRNA splicing to subvert immunity.

The process of RNA splicing influences many physiological processes, including plant immunity. However, how plant parasites manipulate host RNA splicing process remains unknown. Here we demonstrate that PsAvr3c, an avirulence effector from oomycete plant pathogen Phytophthora sojae, physically binds to and stabilizes soybean serine/lysine/arginine-rich proteins GmSKRPs. The SKRPs are novel proteins that associate with a complex that contains plant spliceosome components, and are negative regulators of plant immunity. Analysis by RNA-seq data indicates that alternative splicing of pre-mRNAs from 401 soybean genes, including defense-related genes, is altered in GmSKRP1 and PsAvr3c overexpressing lines compared to control plants. Representative splicing events mediated by GmSKRP1 and PsAvr3c are tested by infection assays or by transient expression in soybean plants. Our results show that plant pathogen effectors can reprogram host pre-mRNA splicing to promote disease, and we propose that pathogens evolved such strategies to defeat host immune systems.

E-cadherin mediates adhesion of Aspergillus fumigatus to non-small cell lung cancer cells.

A spergillus fumigatus is a widely distributed microorganism, and recently, A. fumigatus culture filtrate has been shown to trigger apoptotic cell death in several human cancer cell lines, including non-small lung cancer (NSCLC) cells, A549. Nevertheless, the molecular adhesion of A. fumigatus to these cancer cells to trigger cell death remains unknown. Here, we knocked down E-cadherin in A549 cells and examined its effects on A. fumigatus. The blastospores of A. fumigatus were incubated with the complete protein extracts from A549 cells, using an affinity purification procedure. Preliminary exploration of E-cadherin-interacting protein on the surface of Aspergillus fumigates was done by immunoprecipitation and mass spectrometry analysis. We found that the adhesion of the blastospores to A549 cells was significantly reduced by E-cadherin suppression in A549 cells, suggesting that E-cadherin of A549 cells may mediate the surface adhesion of A. fumigatus blastospore. Mass spectrometry (MS) analysis predicted two binding proteins for E-cadherin on A. fumigatus, AfA24A6.130c and XP_747789. Finally, the growth of E-cadherin-depleted A549 cells significantly increased by infection of A. fumigatus in vivo. Thus, our study suggests that E-cadherin mediates adhesion of A. fumigatus to NSCLC cells to trigger cell death and provides molecular evidence for the treatment of NSCLC with controlled A. fumigatus infection.

[Clinical analysis of 108 cases with adenocarcinoma Barretts's esophagus].

OBJECTIVE: To investigate the prognostic factors and to analyze the efficacy of chemotherapy and/or radiotherapy for Barrett's esophageal adenocarcinoma after radical surgical resection. METHODS: The clinical data of 108 patients with adenocarcinoma Barrett's esophagus picking out from 783 esophageal adenocarcinoma patients surgically treated between June 1978 to June 2001 in the Shandong Provincial Hospital and Shandong Qianfoshan Hospital were analyzed retrospectively. 60Co gamma-irradiation or 6MVX-ray with conventional fraction were used for radiotherapy with a total volume dosage of 55-70 Gy. The chemotherapy was either FAM (iv infusion of 5-Fu 500 mg, d1-d5; ADM 50 mg d1; MMC 12 mg, d1) or CMF regimen (iv infusion of CTX 800 mg d1, d8; MTX 30 mg d1; 5-Fu 500 mg, d1-d5) for 4-6 cycles. The Kaplan-Meier amalysis was used to estimate the survival rate. Log rank test was used for comparison of the survival difference among different groups. RESULTS: In this series, 76 of 92 patients who underwent radical surgical resection received postoperative radiotherapy alone, and 16 received radiotherapy plus chemotherapy. Twelve of the other 16 patients who underwent palliative surgical resection received chemotherapy plus radiotherapy, the remaining 4 patients died of operative complications during surgery. The overall 1-, 3- and 5-year survival rate of this series was 81.5%, 51.9% and 22.2%, respectively. In the radical resection group, it was 15.8% for the patients received radiotherapy alone versus 75.0% for those treated by chemotherapy plus radiotherapy. The 5-year survival rate was 33.3% for the patients without extra-esophageal infiltration and 33.3% for the patients without lymph node metastasis, respectively. However, it was only 9.1% for the patients with extra-esophageal infiltration and 14.3% for those with lymph node metastasis, respectively. For the patients who had palliative surgical resection, though they received chemotherapy plus radiotherapy postoperatively, none of them survived longer than 5-year. Statistically significant difference among these groups was demonstrated by Log rank test (P < 0.05). CONCLUSION: Chemotherapy plus radiotherapy after radical surgical resection may improve the survival of patients with adenocarcinoma in Barrett's esophagus adenocarcinoma patient. The pathological stage, extra-esophageal infiltration, lymph node metastasis and postoperative chemotherapy plus radiotherapy are important prognostic factors.