Ratiometric electrochemical immunoassay based on 2D Co/Fe MOF decorated with toluidine blue and Fc-labeled Schiff base for accurate assay of alpha-fetoprotein in clinical serum.

The high level of alpha-fetoprotein (AFP) expression is closely related to hepatocellular carcinoma (HCC). Herein, a dual signal ratiometric electrochemical immunosensor based on chitosan-ferrocenecarboxaldehyde-spindle gold (Chit-Fc-SAu) and Co/Fe metal-organic framework-toluidine blue/polydopamine (Co/Fe MOF-TB/PDA) was proposed for quantitative analysis of AFP. Specifically, Chit-Fc-SAu worked as a substrate to trap more primary antibodies (Ab1) generating the first electrochemical signal from Fc. Thanks to the large specific surface area, the synergistic and electronic effects of Co/Fe MOF nanosheets, and the rich functional groups of PDA, Co/Fe MOF-TB/PDA could load more secondary antibodies (Ab2) and signal molecules (TB) providing another amplified electrochemical signal. In the presence of AFP, Ab1-AFP-Ab2 formed a sandwich structure, and as the AFP concentration increased, the peak current ratio of TB to Fc (ITB/IFc) also increased. The dual signal ratiometric strategy can avoid environmental signal interference and achieve signal self-calibration, thereby improving the accuracy and reproducibility of detection. After a series of exploration, this self-calibrated ratiometric immunosensor exhibited a wide linear range (0.001-200 ng mL-1), a low detection limit (0.34 pg mL-1), and good repeatability. When applied to the assay of clinical serum samples, the detection results of ratiometric sensor were consistent with that of commercial electrochemiluminescence (ECL) immunoassay, significantly superior to that of non-ratiometric sensor. The self-calibrated strategy based on ratiometric sensor helps to improve the accuracy of AFP in clinical diagnosis.

Facile preparation of Ru nanoassemblies for electrochemical immunoassay of carcinoembryonic antigen in clinical serum.

Abnormal expression of carcinoembryonic antigen (CEA) can be used for early diagnosis of various cancers (e.g. colorectal cancer, cervical carcinomas, and breast cancer). In this work, using l-cysteine-ferrocene-ruthenium nanocomposites (L-Cys-Fc-Ru) to immobilize secondary antibody (Ab2) and Au nanoparticles (NPs) as the substrate to ensure accurate capture of primary antibody (Ab1), a signal-on sandwich-like biosensor was constructed in the presence of CEA. Specifically, Ru nanoassemblies (NAs) were first prepared by a facile one-step solvothermal approach as signal amplifiers for the electrical signal of Fc. Based on specific immune recognition, as the increase of CEA concentration, the content of L-Cys-Fc-Ru-Ab2 captured on the electrode surface also increased, thus the signal of Fc gradually increased. Therefore, the quantitative detection of CEA can be realized according to the peak current of Fc. After a series of experiments, it was found that the biosensor has a wide detection range from 1.0 pg mL-1 to 100.0 ng mL-1 and a low detection limit down to 0.5 pg mL-1, as well as good selectivity, repeatability and stability. Furthermore, satisfactory results were also obtained for the determination of CEA in serums, which were comparable to commercial electrochemiluminescence (ECL) method. The developed biosensor shows great potential in clinical applications.

Simple synthesis of PtRu nanoassemblies as signal amplifiers for electrochemical immunoassay of carbohydrate antigen 19-9.

In clinical analysis, carbohydrate antigen 19-9 (CA199) is a gold standard for pancreatic cancer diagnosis. Herein, PtRu nanoassemblies (NAs) were synthesized via a facile one-step solvothermal approach, with the help of octylphenoxypolye thoxyethanol (NP-40) acted as a growth-directing molecule, and triethylene glycol (TEG) worked as a reductant and solvent. During the assembly process of small particles, a large number of voids were formed, which significantly increase the specific surface area of the PtRu NAs exhibiting excellent electrocatalytic performance. Incorporating the PtRu NAs as signal amplifiers for potassium ferrocyanide oxidation into the specific molecular recognition of proteins, a facile signal-enhanced electrochemical (EC) immunosensor was developed. Verified by a series of experiments, the proposed immunosensor presented a wide linear range (10-4-70 U mL-1) and a low detection limit (3.3 × 10-5 U mL-1), accompanied by good reproducibility, selectivity, and stability, which could be applied in human serum samples for the determination of CA199, and was comparable to commercial electrochemiluminescence (ECL) immunoassay. Feasibility of batch fabrication of PtRu NAs makes nanomaterial-based EC immunoassay promising for the determination of similar cancer markers in future.

Preservation affects the vegetative growth and fruiting body production of Cordyceps militaris.

Cordyceps militaris is a model species of Cordyceps fungi, and has been traditionally used as an edible and medicinal fungus due to its richness of bioactive and pharmacological metabolites. The fruiting bodies of this fungus are widely used as healthy food and nutrition supply. In industrial production, fruiting bodies are cultivated on artificial media, but their yield and quality are usually affected by the quality of fungal strains. In this study, the effect of colony growth rate of the fungal strains, fungal age and repeated subculturing on the fungal biomass accumulation was investigated. The results indicated that the fungal biomass was positively correlated with the colony growth rate and not affected by fungal age and the repeated subculturing. The preservation conditions for stock cultures, including choice of cultures, lyophilization, temperature and protective agents were optimized based on the mycelial formation and conidia production in artificial inoculum. The development of fruiting bodies from the fungal strains stored under the optimized preservation conditions were further analyzed to determine the ideal time period of preservation. Results indicated that storing the fungus at 4 °C could maintain the fungal vitality and fruiting body producing capacity for at least 12 months. This study established practical criteria of fungal inoculum for artificial cultivation of fruiting body and provided a simple and efficient preservation method for C. militaris. The results may shed light on preservation for other Cordyceps species and other edible fungi.

Manipulating Depletion Region of Aqueous-Processed Nanocrystals Solar Cells with Widened Fermi Level Offset.

Water soluble nanocrystals (NCs) are promising materials in aqueous-processed solar cells because of their high extinction coefficient, low-cost, and favorable photoelectric characteristics. However, the power conversion efficiency (PCE) of the present aqueous-processed NC solar cells is restricted by the short depletion region of the active layer and limited Fermi level offset between NCs and the electron transport layer. Herein, these issues are effectively addressed by preparing Cdx Zn1- x Te NCs capped with 2-aminoethanethiol hydrochloride. The introduction of Zn2+ into CdTe NCs widens the Fermi level offset from 0.68 to 0.74 eV, lengthens the depletion region from 130 to 137 nm, and hence brings obvious improvement in the open circuit voltage (Voc ) and fill factor. Especially, the depletion region is successfully tuned from 137 to 171 nm, and even lengthened to a record thickness of 200 nm based on aqueous-processed solar cells. As a result, a champion thickness ratio (74%) of depletion region to active layer (200/270 nm) is achieved. A champion PCE of 5.96% and short-circuit current (Jsc ) of 21.2 mA cm-2 are achieved among aqueous-processed NC solar cells. This work provides a simple way to prepare polynary NCs and highlights a prospective method to develop more efficient and cost-effective solution-processed environment friendly solar cells.

Temporal Change of the Content of 10 Oligosaccharides in the Milk of Chinese Urban Mothers.

Breastfed infants tend to be less prone to infections and may have improved cognitive benefits compared to formula-fed infants. Human milk oligosaccharides (HMO) are the third most abundant component of human milk, but are absent from formulae. They may be partially responsible for the benefits of breastfeeding. In this cross-sectional observational study, the HMO composition of milk from Chinese mothers was studied to determine the impact of stage of lactation, mode of delivery and geographical location. The content of 10 HMO was measured by HPLC in 446 milk samples from mothers living in three different cities in China. Around 21% of the samples contained levels of 2'-fucosyllactose (2'-FL) below the limit of quantification, which is similar to the frequency of fucosyltransferase-2 non-secretors in other populations, but 2'-FL was detected in all samples. Levels of most of the HMO studied decreased during the course of lactation, but the level of 3-fucosyllactose increased. Levels of 2'-FL and 3-fucosyllactose seem to be strongly correlated, suggesting some sort of mechanism for co-regulation. Levels of 6'-sialyllactose were higher than those of 3'-sialyllactose at early stages of lactation, but beyond 2-4 months, 3'-sialyllactose was predominant. Neither mode of delivery nor geographical location had any impact on HMO composition.

RGD-modified endostatin peptide 30 derived from endostatin suppresses invasion and migration of HepG2 cells through the αvß3 pathway.

The nucleotide sequence encoding amino acids 1-30 of endostatin (peptide 30, with amino acids 25-31 mutated from RGIRGAD to RGDRGD) was artificially synthesized and cloned into the plasmid pTYB2 and expressed in Escherichia coli (DE3). Peptide 30 was purified by chitin affinity chromatography followed by dithiothreitol removal by gel filtration and protein identification using Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Similarly, peptide 27, corresponding to amino acids 1-27 of endostatin, was produced as control. The effects of peptide 30 on the metastatic potential of HepG2 cells were then investigated. Peptide 30 was found to effectively suppress the adhesion, invasion, and migration of HepG2 cells. Flow cytometry demonstrated that peptide 30 did not alter the expression of membrane integrin αvß3, although immunofluorescent staining revealed that these integrins formed clusters on the cell surface. Therefore, the effects of peptide 30 on cancer cell invasion may involve the αvß3 pathway. Moreover, peptide 30 inactivated metalloproteinase-2 (MMP-2) and MMP-9 and downregulated the expression of COX-2 (cyclooxygenase 2), MMP-2, and MMP-9 at both mRNA and protein levels. Peptide 30 also upregulated the expression of tissue inhibitor of metalloproteinase-1 and tissue inhibitor of metalloproteinase-2 at mRNA and protein levels. Lastly, an antibody against αvß3 enhanced the biological effects of peptide 30.