Physicochemical Properties of Plasma-Activated Water and Its Control Effects on the Quality of Strawberries.

In this study, the effects of plasma-activated water (PAW), generated by dielectric barrier discharge cold plasma at the gas-liquid interface, on the quality of fresh strawberries during storage were investigated. The results showed that, with the prolongation of plasma treatment time, the pH of PAW declined dramatically and the electrical conductivity increased significantly. The active components, including NO2-, NO3-, H2O2, and O2-, accumulated gradually in PAW, whereas the concentration of O2- decreased gradually with the treatment time after 2 min. No significant changes were found in pH, firmness, color, total soluble solids, malondialdehyde, vitamin C, or antioxidant activity in the PAW-treated strawberries (p > 0.05). Furthermore, the PAW treatment delayed the quality deterioration of strawberries and extended their shelf life. Principal component analysis and hierarchical cluster analysis showed that the PAW 2 treatment group demonstrated the best prolonged freshness effect, with the highest firmness, total soluble solids, vitamin C, and DPPH radical scavenging activity, and the lowest malondialdehyde and ∆E* values, after 4 days of storage. It was concluded that PAW showed great potential for maintaining the quality of fresh fruits and extending their shelf life.

Single-Atom Ruthenium Biomimetic Enzyme for Simultaneous Electrochemical Detection of Dopamine and Uric Acid.

Single-atom catalysts have attracted numerous attention due to the high utilization of metallic atoms, abundant active sites, and highly catalytic activities. Herein, a single-atom ruthenium biomimetic enzyme (Ru-Ala-C3N4) is prepared by dispersing Ru atoms on a carbon nitride support for the simultaneous electrochemical detection of dopamine (DA) and uric acid (UA), which are coexisting important biological molecules involving in many physiological and pathological aspects. The morphology and elemental states of the single-atom Ru catalyst are studied by transmission electron microscopy, energy dispersive X-ray elemental mapping, high-angle annular dark field-scanning transmission electron microscopy, and high-resolution X-ray photoelectron spectroscopy. Results show that Ru atoms atomically disperse throughout the C3N4 support by Ru-N chemical bonds. The electrochemical characterizations indicate that the Ru-Ala-C3N4 biosensor can simultaneously detect the oxidation of DA and UA with a separation of peak potential of 180 mV with high sensitivity and excellent selectivity. The calibration curves for DA and UA range from 0.06 to 490 and 0.5 to 2135 µM with detection limits of 20 and 170 nM, respectively. Moreover, the biosensor was applied to detect DA and UA in real biological serum samples using the standard addition method with satisfactory results.

Portable photoelectrochemical immunoassay with micro-electro-mechanical-system for alpha-fetoprotein in hepatocellular carcinoma.

Early detection of cancer has a profound impact on patient survival and treatment outcomes considering high treatment success rates and reduced treatment complexity. Here, we developed a portable photoelectrochemical (PEC) immune platform for sensitive testing of alpha-fetoprotein (AFP) based on Pt nanocluster (Pt NCs) loaded defective-state g-C3N4 photon-electron transducers. The broad forbidden band structure of g-C3N4 was optimized by the nitrogen doping strategy and additional homogeneous porous structure was introduced to further enhance the photon utilization. In addition, the in-situ growth of Pt NCs provided efficient electron transfer catalytic sites for sacrificial agents, which were used to further improve the sensitivity of the sensor. Efficient photoelectric conversion under a hand-held flashlight was determined by the geometry of the transducer and the energy band design, and the portable design of the PEC sensor was realized. The developed sensing platform exhibited a wide linear response range (0.1-50 ng mL-1) and low limit of detection (0.043 ng mL-1) for AFP under optimum conditions. This work provides a new idea for designing portable PEC biosensing platforms to meet the current mainstream POC testing needs.

Liposome-embedded PtCu nanozymes for improved immunoassay of accurate myocardial infarction.

Herein, we developed a platinum-copper nano-enzyme-linked immunosorbent assay (NLISA) based split diagnostic platform for the ultrasensitive detection of cardiac troponin I (cTnI). The PtCu nanozyme synthesized by one-pot synthesis exhibited ultra-high peroxidase-like activity (35.17 U mg-1), which was about 4.5 times higher than that of the unmodified Pt nanozyme (8.83 U mg-1). Due to the efficient peroxidase-like activity of the copper-platinum complexed nanozyme, transduction and sequential amplification of cTnI biological signals were achieved in combination with a liposome-embedded amplification strategy. The encapsulation efficiency was calculated by introducing a liposomal bilayer model, which showed that the introduction of a single liposomal molecule could amplify the signal up to 870-fold, thus promising a high sensitivity test. Notably, the dynamic response of cTnI was in the range of 0.1-5000 pg mL-1 with an ultra-low detection limit (0.048 pg mL-1). The developed NLISA analysis system provides a new way to discover efficient and sensitive alternatives to ELISA kits, which can meet the practical needs of community healthcare testing conditions and rapid testing in hospitals.

Highly selective fluorescence detection of L-selenium-methylselenocysteine in selenium-enriched Cardamine violifolia.

A feasible and practicable "off-on" type of fluorescence strategy for highly selective screening of L-selenium-methylselenocysteine (L-SeMC) in selenium-enriched Cardamine violifolia was developed using g-C3N4-MnO2 nanocomposites as fluorescent probes. The g-C3N4 nanosheets can emit blue fluorescence at 320 nm excitation wavelength with a fluorescence quantum yield of 28%. When MnO2 was deposited onto g-C3N4 nanosheets, the fluorescence of the g-C3N4 nanosheets was quenched due to fluorescence resonance energy transfer (FRET). After the addition of L-SeMC, MnO2 was reduced to Mn2+, which eliminated FRET and fluorescence was restored. Based on this, a quantitative method for the determination of L-SeMC was established. The fluorescence intensity of g-C3N4-MnO2 nanocomposites showed a good linear relationship with the concentration of L-SeMC in the range of 0-45 µg mL-1, the limit of detection (LOD, 3σ/K) was 8.25 ng mL-1 and the correlation coefficient was 0.9904. Common selenium compounds such as SeO2, Na2SeO3, SeMet and SeCys caused weak fluorescence intensity, which means that the developed method is highly selective to detect L-SeMC in a series of selenium compounds. Meanwhile, the technique was evaluated by spiking L-SeMC standards in C. violifolia extraction solutions and with 9 C. violifolia extraction specimens, receiving excellent accordance with results from the commercially available atomic fluorescence spectroscopy method.

Response of Phyllosphere and Rhizosphere Microbial Communities to Salt Stress of Tamarix chinensis.

As carriers of direct contact between plants and the atmospheric environment, the microbiomes of phyllosphere microorganisms are increasingly recognized as an important area of study. Salt secretion triggered by salt-secreting halophytes elicits changes in the community structure and functions of phyllosphere microorganisms, and often provides positive feedback to the individual plant/community environment. In this study, the contents of Na+ and K+ in the rhizosphere, plant and phyllosphere of Tamarix chinensis were increased under 200 mmol/L NaCl stress. The increase in electrical conductivity, Na+ and K+ in the phyllosphere not only decreased the diversity of bacterial and fungal communities, but also decreased the relative abundance of Actinobacteriota and Basidiomycota. Influenced by electrical conductivity and Na+, the bacteria-fungus co-occurrence network under salt stress has higher complexity. Changes in the structure of the phyllosphere microbial community further resulted in a significant increase in the relative abundance of the bacterial energy source and fungal pathotrophic groups. The relative abundance of Actinobacteriota and Acidobacteriota in rhizosphere showed a decreasing trend under salt stress, while the complexity of the rhizosphere co-occurrence network was higher than that of the control. In addition, the relative abundances of functional groups of rhizosphere bacteria in the carbon cycle and phosphorus cycle increased significantly under stress, and were significantly correlated with electrical conductivity and Na+. This study investigated the effects of salinity on the structure and physicochemical properties of phyllosphere and rhizosphere microbial communities of halophytes, and highlights the role of phyllosphere microbes as ecological indicators in plant responses to stressful environments.

Development of a novel fluorescent protein-based probe for efficient detection of Pb2+ in serum inspired by the metalloregulatory protein PbrR691.

BACKGROUND: The accurate and rapid detection of blood lead concentration is of paramount importance for assessing human lead exposure levels. Fluorescent protein-based probes, known for their high detection capabilities and low toxicity, are extensively used in analytical sciences. However, there is currently a shortage of such probes designed for ultrasensitive detection of Pb2+, and no reported probes exist for the quantitative detection of Pb2+ in blood samples. This study aims to fill this critical void by developing and evaluating a novel fluorescent protein-based probe that promises accurate and rapid lead quantification in blood. RESULTS: A simple and small-molecule fluorescent protein-based probe was successfully constructed herein using a peptide PbrBD designed for Pb2+ recognition coupled to a single fluorescent protein, sfGFP. The probe retains a three-coordinate configuration to identify Pb2+ and has a high affinity for it with a Kd' of 1.48 ± 0.05 × 10-17 M. It effectively transfers the conformational changes of the peptide to the chromophore upon Pb2+ binding, leading to fast fluorescence quenching and a sensitive response to Pb2+. The probe offers a broad dynamic response range of approximately 37-fold and a linear detection range from 0.25 nM to 3500 nM. More importantly, the probe can resist interference of metal ions in living organisms, enabling quantitative analysis of Pb2+ in the picomolar to millimolar range in serum samples with a recovery percentage of 96.64%-108.74 %. SIGNIFICANCE: This innovative probe, the first to employ a single fluorescent protein-based probe for ultrasensitive and precise analysis of Pb2+ in animal and human serum, heralds a significant advancement in environmental monitoring and public health surveillance. Furthermore, as a genetically encoded fluorescent probe, this probe also holds potential for the in vivo localization and concentration monitoring of Pb2+.

Cerium oxide-triggered 'one-to-many' catalytic cycling strategy for in situ amplified electronic signal of low-abundance protein.

Multifunctionalized thionine-modified cerium oxide (Thi-CeO2) nanostructures with redox ability and catalytic activity were designed as the bionanolabels for in situ amplified electronic signal of low-abundance protein (carcinoembryonic antigen, CEA, used as a model) based on a cerium oxide-triggered 'one-to-many' catalytic cycling strategy. Initially, the carried CeO2 nanoparticles autocatalytically hydrolyzed the phosphate ester bond of l-ascorbic acid 2-phosphate (AAP) to produce a new reactant (l-ascorbic acid, AA), then the generated AA was electrochemically oxidized by the assembled thionine on the Thi-CeO2, and the resultant product was then reduced back to AA by the added tris(2-carboxyethy)phosphine (TCEP). The catalytic cycling could be re-triggered by the thionine and TCEP, resulting in amplification of the electrochemical signal. Under the optimized conditions, the electrochemical immunosensor exhibited a wide linear range of 0.1 pg mL(-1) to 80 ng mL(-1) with a low detection limit of 0.08 pg mL(-1) CEA at the 3σblank level. In addition, the methodology was evaluated for the analysis of clinical serum samples, and was in good accordance with values obtained using the commercialized enzyme-linked immunosorbent assay (ELISA) method.

Study on the association between adverse drug reactions to opioids and gene polymorphisms: a case-case-control study.

OBJECTIVE: Adverse drug reactions (ADRs) caused by opioid drugs show individual differences. Our objective was to explore the association between gene polymorphism and ADRs induced by opioid drugs. METHODS: Evidence-based medical data analysis was conducted for genes related to ADRs induced by opioid drugs to select target genes. Sixty patients with cancer pain who had ADRs after taking opioid drugs (morphine, codeine, oxycodone) and 60 patients without ADRs after taking opioid drugs were used as the experimental group and control group, respectively. Then, we used polymerase chain reaction (PCR) or in situ hybridization to detect target genes. By combining with clinical data such as age, sex, dosage and duration of medication, the effect of gene polymorphism on the ADR of patients after taking opioid drugs was statistically analysed. RESULTS: Based on a database search and evidence-based medical data, we identified CYP2D6*10, CYP3A5*3, ABCB1, and OPRM1 as target genes for detection. The results of statistical analysis showed no significant difference in genotype distribution between the experimental group and the control group (p > 0.05). However, if 32 patients with ADRs after taking oxycodone and 32 controls were selected for comparison, the SPSS22.0 and SNPStats genetic models showed that the ABCB1 (062rs1045642) CT and TT genotypes correlated with the occurrence of ADRs (p < 0.05): the total number of CT + TT genotypes in the experimental group was 29 (90.62%), with 11 (34.37%) CT + TT genotypes types in the control group. CONCLUSION: Polymorphism of ABCB1 (062rs1045642) is related to ADRs caused by oxycodone, and the incidence of ADRs is higher with the allele T. Polymorphism of ABCB1 is expected to become a clinical predictor of ADRs to oxycodone, and attention should be given to the occurrence of serious ADRs in patients with ABCB1 (062rs1045642) CT and TT genotypes.

Ultrasensitive aptamer-based multiplexed electrochemical detection by coupling distinguishable signal tags with catalytic recycling of DNase I.

This work reports an aptamer-based, disposable, and multiplexed sensing platform for simultaneous electrochemical determination of small molecules, employing adenosine triphosphate (ATP) and cocaine as the model target analytes. The multiplexed sensing strategy is based on target-induced release of distinguishable redox tag-conjugated aptamers from a magnetic graphene platform. The electronic signal of the aptasensors could be further amplified by coupling DNase I with catalytic recycling of self-produced reactants. The assay was based on the change in the current at the various peak potentials in the presence of the corresponding signal tags. Experimental results revealed that the multiplexed electrochemical aptasensor enabled the simultaneous monitoring of ATP and cocaine in a single run with wide working ranges and low detection limits (LODs: 0.1 pM for ATP and 1.5 pM for cocaine). This concept offers promise for rapid, simple, and cost-effective analysis of biological samples.

An organic-inorganic hybrid nanostructure-functionalized electrode for electrochemical immunoassay of biomarker by using magnetic bionanolabels.

A new electrochemical immunoassay of alpha-fetoprotein (AFP) was developed on an organic-inorganic hybrid nanostructure-functionalized carbon electrode by coupling with magnetic bionanolabels. Multi-walled carbon nanotubes (CNTs), single-stranded DNA, thionine and AFP were utilized for the construction of the immunosensor, while the core-shell Fe(3)O(4)-silver nanocomposites were employed for the label of horseradish peroxidase-anti-AFP conjugates (HRP-anti-AFP-AgFe). Electrochemical measurement toward AFP was carried out by using magnetic bionanolabels as traces and H(2)O(2) as enzyme substrate with a competitive-type immunoassay mode. Experimental results indicated that the immunosensors with carbon nanotubes and DNA exhibited better electrochemical responses than those of without carbon nanotubes or DNA. Under optimal conditions, the electrochemical immunosensor by using HRP-anti-AFP-AgFe as signal antibodies exhibited a linear range of 0.001-200 ng mL(-1) AFP with a low detection limit of 0.5 pg mL(-1) at 3s(B). Both intra- and inter-assay coefficients of variation were 7.3%, 9.4%, 8.7% and 10.2%, 7.8%, 9.4% toward 0.01, 30, 120 ng mL(-1) AFP, respectively. The specificity and stability of the electrochemical immunoassay were acceptable. In addition, the methodology was validated for 12 clinical serum specimens including 9 positive specimens and 3 normal specimens, receiving a good correlation with the results obtained from the referenced electrochemiluminescence assay.

A graphene-based Au(111) platform for electrochemical biosensing based catalytic recycling of products on gold nanoflowers.

A novel, enzyme-free amperometric immunoassay of biomarkers with sensitive enhancement was designed by using gold nanoflower-labeled detection antibodies toward the catalytic reduction of p-nitrophenol and redox cycling of p-aminophenol on a graphene-based Au(111) platform.

Specific immunoreaction-induced controlled release strategy for sensitive impedance immunoassay of a cancer marker.

A simple and facile impedance immunoassay strategy for sensitive detection of alpha-fetoprotein (AFP), as a model cancer marker, was developed by using target-induced release of nanogold particle-labelled anti-AFP antibodies from polyvinylpyrrolidone-coated magnetic carbon nanotubes.

YKL-40 expression in human hepatocellular carcinoma: a potential biomarker?

BACKGROUND: YKL-40 is a new biomarker with diagnostic value in many different cancers. Whether it may serve as a biomarker for hepatocellular carcinoma (HCC) is still unclear. This study aimed to examine the expression of YKL-40 in the serum and liver tissues of HCC patients and in HCC cell lines, in comparison with that in non-HCC liver disease patients and non-tumor hepatic cell lines, respectively. METHODS: Immunohistochemical staining was used to detect YKL-40 protein expression in liver biopsy specimens from 8 HCC patients. ELISA was used to assess the serum YKL-40 level in 90 HCC patients, 90 inactive HBsAg carrier (IHC) patients with normal liver functions, and 90 liver cirrhosis patients. Real-time PCR was used to determine the YKL-40 mRNA expression in three HCC cell lines and two non-tumor hepatic cell lines. RESULTS: Immunohistochemical staining of liver biopsy specimens from HCC patients showed that the YKL-40 protein expression in tumor tissue was higher than that in adjacent normal tissues. ELISA revealed that the YKL-40 serum level in the HCC group was significantly higher than that in the IHC group, but not significantly different from that in the cirrhosis group. Real-time PCR showed that YKL-40 mRNA levels in HCC cell lines were significantly higher than those in non-tumor hepatic cells. CONCLUSIONS: YKL-40 is highly expressed in HCC at the molecular, cellular and tissue levels. However, it may not serve as a serum biomarker for HCC because measurement of the serum YKL-40 level cannot distinguish HCC from cirrhosis.

Targeting human epidermal growth factor receptor signaling with the neuregulin's heparin-binding domain.

A major limitation in biopharmaceutical development is selectively targeting drugs to diseased tissues. Growth factors and viruses have solved this problem by targeting tissue-specific cell-surface heparan sulfates. Neuregulin (NRG), a growth factor important in both nervous system development and cancer, has a unique heparin-binding domain (HBD) that targets to cell surfaces expressing its HER2/3/4 receptors (Esper, R. M., Pankonin, M. S., and Loeb, J. A. (2006) Brain Res. Rev. 51, 161-175). We have harnessed this natural targeting ability of NRG by fusing the HBD of NRG to soluble HER4. This fusion protein retains high affinity heparin binding to heparin and to cells that express heparan sulfates resulting in a more potent NRG antagonist. In vivo, it is targeted to peripheral nerve segments where it blocks the activity of NRG as a Schwann cell survival factor. The fusion protein also efficiently blocks autocrine and paracrine signaling and reduces the proliferation of MCF10CA1 breast cancer cells. These findings demonstrate the utility of the HBD of NRG in biopharmaceutical targeting and provide a new way to block HER signaling in cancer cells.

A Rare Case of the Newly Recognized Kaposi Sarcoma Herpesvirus-Associated Disease.

Kaposi sarcoma herpesvirus (KSHV) is associated with Kaposi sarcoma (KS), primary effusion lymphoma, and multicentric Castleman disease (KSHV-MCD) in patients infected with human immunodeficiency virus (HIV). We present a case consistent with a newly recognized KSHV inflammatory cytokine syndrome (KICS), distinct from KSHV-MCD. Although both disorders exhibit signs of substantial inflammation, KICS has minimal lymphadenopathy/splenomegaly and negative pathologic nodal changes in the setting of low CD4 count. KICS is easily misdiagnosed as severe sepsis or other KS-related diseases in HIV/AIDS patients and carries a high mortality. ​Standard therapy is still under investigation due to its rarity, whereas the treatment regimen for KSHV-MCD may lead to clinical remission. Early recognition and prompt management are crucial to improve the survival of the under-recognized KICS.

Interferon regulatory factors IRF5 and IRF7 inhibit growth and induce senescence in immortal Li-Fraumeni fibroblasts.

Cellular immortalization is one of the prerequisite steps in carcinogenesis. By gene expression profiling, we have found that genes in the interferon (IFN) pathway were dysregulated during the spontaneous cellular immortalization of fibroblasts from Li-Fraumeni syndrome (LFS) patients with germ-line mutations in p53. IFN signaling pathway genes were down-regulated by epigenetic silencing during immortalization, and some of these same IFN-regulated genes were activated during replicative senescence. Bisulfite sequencing of the promoter regions of two IFN regulatory transcription factors (IRF5 and IRF7) revealed that IRF7, but not IRF5, was epigenetically silenced by methylation of CpG islands in immortal LFS cells. The induction of IRF7 gene by IFNalpha in immortal LFS cells was potentiated by pretreatment with the demethylation agent 5-aza-2'-deoxycytidine. Overexpression of IRF5 and IRF7 revealed that they can act either alone or in tandem to activate other IFN-regulated genes. In addition, they serve to inhibit the proliferation rate and induce a senescence-related phenotype in immortal LFS cells. Furthermore, polyinosinic:polycytidylic acid treatment of the IRF-overexpressing cells showed a more rapid induction of several IFN-regulated genes. We conclude that the epigenetic inactivation of the IFN pathway plays a critical role in cellular immortalization, and the reactivation of IFN-regulated genes by transcription factors IRF5 and/or IRF7 is sufficient to induce cellular senescence. The IFN pathway may provide valuable molecular targets for therapeutic interventions at early stages of cancer development.

Short-term entecavir therapy of chronic severe hepatitis B.

BACKGROUND: Chronic severe hepatitis B patients often have limited survival. This investigation aimed to evaluate the short-term effects of nucleoside analog therapy on chronic severe hepatitis B. METHODS: We retrospectively, randomly collected the data of 129 chronic severe hepatitis B patients: 55 were treated with entecavir, and the remaining 74 were not treated with nucleoside analogues. RESULTS: No significant difference in short-term survival rate was found between the group treated with entecavir and that treated without nucleoside analogues. Although entecavir greatly reduced HBV replication in different periods of therapy (P<0.001), the model for end-stage liver disease (MELD) score and liver function (alanine aminotransferase, albumin, bilirubin, prothrombin time) showed no significant change. No significant differences were found in MELD scores and liver function in patients with different HBV DNA levels (< or =10(4) copies/ml, >10(4) to <10(6) copies/ml, > or =10(6) copies/ml). Nor correlation was observed between HBV DNA levels and MELD scores in different periods of therapy (P>0.05). The HBV DNA levels of patients who survived for over 3 months or less than 3 months were not significantly different either. However, the MELD score and parameters of liver function (albumin, bilirubin, prothrombin time) were different between the two groups (P<0.05). CONCLUSION: These results suggest that short-term suppression of HBV replication may not slow down the progression of liver failure in patients with chronic severe hepatitis B.

Inequalities for the fractional convolution operator on differential forms.

The purpose of this paper is to derive some Coifman type inequalities for the fractional convolution operator applied to differential forms. The Lipschitz norm and BMO norm estimates for this integral type operator acting on differential forms are also obtained.

Stat1 expression is not sufficient to regulate the interferon signaling pathway in cellular immortalization.

DNA hypermethylation in gene promoters is an epigenetic mechanism regulating gene expression in cellular immortalization, an important step in carcinogenesis. Previously, we studied the genes dysregulated during immortalization using spontaneously immortalized fibroblasts from patients with Li-Fraumeni syndrome (LFS), who carry a germline mutation in the tumor suppressor gene p53. We found that multiple interferon (IFN) signaling pathway genes were regulated by epigenetic silencing. In this study we focused on a key regulator of that pathway, the signal transducer and transcription activator 1 (Stat1) gene. Although Stat1 is downregulated after cellular immortalization and upregulated in immortal MDAH041 cells after 5-aza-2'-deoxycytidine (5-aza-dC) treatment, we detected no methylation of the Stat1 promoter region in these cells before or after immortalization. To analyze the function of Stat1 in immortalization, we expressed Stat1 in immortal MDAH041 cells by stable infection, expecting to induce IFN-regulated genes or cellular senescence or both. However, the overexpression of Stat1 alone was not sufficient to repress the proliferation rate of immortal MDAH041 cells or induce senescence in immortal MDAH041 cells. We concluded that factor(s) additional to Stat1 (whether IFN dependent or not) are required for the immortalization of LFS fibroblasts.