A Pt-Ir nanocube amplified lateral flow immunoassay for dehydroepiandrosterone.

The traditional gold-nanoparticle-based lateral flow immunoassay (LFIA) cannot satisfy the requirements for the sensitive detection of dehydroepiandrosterone (DHEA) in human urine. To enhance the sensitivity of the LFIA, platinum-iridium nanocubes (Pt-Ir NCs) with high catalytic efficiency and stability were synthesized and labelled with polyclonal antibody (pAb) to form a pAb-Pt-Ir probe. For the detection of DHEA, a novel LFIA with Pt-Ir NCs as an optical label and an enhanced LFIA in which the peroxidase-like activity of the Pt-Ir NCs was triggered by the introduction of the chromogenic substrate 3-amino-9-ethyl-carbazole (AEC) were developed and compared with a LFIA with platinum nanocubes (PtNCs) as an optical label. The visual limit of detection was 0.5 ng mL-1 for Pt-Ir-LFIA and 0.05 ng mL-1 for AEC-enhanced Pt-Ir-LFIA, in comparison to 100 ng mL-1 for PtNCs-LFIA and 50 ng mL-1 for AEC-enhanced PtNCs-LFIA. The average recoveries from spiked urine samples ranged from 90.8% to 110.4%, with a coefficient of variation below 12.6%, suggesting the accuracy and reliability of our developed immunoassay. Achieving excellent sensitivity, specificity, and reproducibility, Pt-Ir-LFIA provided a promising platform for monitoring DHEA.

A bifunctional immunosensor based on osmium nano-hydrangeas as a catalytic chromogenic and tinctorial signal output for folic acid detection.

As a neglected member of the platinum group elements, osmium, the metal with the highest density in the earth, is very suitable for the preparation of a peroxidase with high catalytic activity and stability, and can also be associated with the development of a sensor. In this study, we accessed Os nano-hydrangeas (OsNHs) with one-pot synthesis and utilized them in a bifunctional immunosensor that can present both catalytic chromogenic and tinctorial signal for nanozyme-linked immunosorbent assay (NLISA) and lateral flow immunoassay (LFIA) for use in folic acid (FA) detection. In the OsNHs-NLISA, the linear range is from 9.42 to 167.53 ng mL-1. The limit of detection (LOD) is 4.03 ng mL-1 and the IC50 value is 39.73 ng mL-1. In OsNHs-LFIA, the visual cut-off value and limit of detection (v-LOD) are 100 ng mL-1 and 0.01 ng mL-1, respectively. Additionally, the outcome from the specificity and spiked sample analysis offered recovery from the spiked milk powder sample ranging from 93.9 to 103.6% with a coefficient of variation under 4.9%, compared with UPLC-MS/MS for a correlation of R2 = 0.999 and admirable validation. The promising application of the OsNHs can also be used in other bioprobes, and this bifunctional immunosensor analysis mode is suitable for diversified analytes.

Fluorescence polarization immunoassay for rapid determination of dehydroepiandrosterone in human urine.

In this paper, five fluorescein-labeled dehydroepiandrosterone (DHEA) derivatives (tracers) with different chain lengths between the fluorescein and hapten were synthesized and featured so as to establish a fluorescence polarization immunoassay (FPIA) for DHEA detection in human urine samples with previously prepared polyclonal antibody against DHEA. The outcomes of the structure of tracer on FPIA sensitivity were investigated. Under the optimal condition, the fluorescence polarization value (FP) decreases linearly in DHEA concentration, ranging from 1.6 to 243.3 ng mL-1, with the limit of detection of 1.1 ng mL-1 and IC50 value of 25.1 ng mL-1. Moreover, the developed FPIA was time-saving as it could complete the detection within 3 min. FPIA and commercial enzyme-linked immunosorbent assay kit were both applied to analyze the spiked human urine samples with DHEA. Excellent recoveries (92.1-108.0%) and satisfactory correlation coefficient (R2 = 0.98) were acquired with the two methods, indicating that the developed FPIA was a fast and efficient screening immunoassay with accuracy and sensitivity for DHEA detection in human urine samples. Graphical abstract.

Effects of Acupotomy on Immobilization-Induced Gastrocnemius Contracture and Fibrosis in Rats via Wnt/ß-Catenin Signaling.

OBJECTIVE: To determine whether acupotomy ameliorates immobilization-induced muscle contracture and fibrosis via Wnt/ß-catenin signaling pathway. METHODS: Thirty Wistar rats were randomly divided into 5 groups (n=6) by a random number table, including control, immobilization, passive stretching, acupotomy, and acupotomy 3 weeks (3-w) groups. The rat model of gastrocnemius contracture was established by immobilizing the right hind limb in plantar flexion for 4 weeks. Rats in the passive stretching group received passive stretching at gastrocnemius, a daily series of 10 repetitions for 30 s each at 30-s intervals for 10 consecutive days. Rats in the acupotomy and acupotomy 3-w groups received acupotomy once and combined with passive stretching at gastrocnemius a daily series of 10 repetitions for 30 s each at 30-s intervals for 10 consecutive days. Additionally, rats in the acupotomy 3-w group were allowed to walk freely for 3 weeks after 10-day therapy. After treatment, range of motion (ROM), gait analysis [i.e., paw area, stance/swing and maximum ratio of paw area to paw area duration (Max dA/dT)], gastrocnemius wet weight and the ratio of muscle wet weight to body weight (MWW/BW) were tested. Gastrocnemius morphometric and muscle fiber cross-sectional area (CSA) were assessed by hematoxylin-eosin staining. Fibrosis-related mRNA expressions (i.e., Wnt 1, ß-catenin, axin-2, α-smooth muscle actin, fibronectin, and types I and III collagen) were measured using real-time quantitative polymerase chain reactions. Wnt 1, ß-catenin and fibronectin concentrations were measured by enzyme-linked immunosorbent assay. Types I and III collagen in the perimysium and endomysium were analyzed using immunofluorescence. RESULTS: Compared with the control group, ROM, gait function, muscle weight, MWW/BW and CSA were significantly decreased in the immobilization group (all P<0.01), while protein levels of types I and III collagen, Wnt 1, ß-catenin, fibronectin and mRNA levels of fibrosis-related genes were obviously increased (all P<0.01). Treatment with passive stretching or acupotomy restored ROM and gait function and increased muscle wet weight, MWW/BW and CSA (all P<0.05), while protein expression levels of Wnt 1, ß-catenin, fibronectin, types I and III collagen and mRNA levels of fibrosis-related genes were remarkably declined compared with the immobilization group (all P<0.05). Compared with passive stretching group, ROM, gait function, MWW was remarkably restored (all P<0.05), and mRNA levels of fibrosis-related genes as well as protein expression levels of Wnt 1, ß-catenin, fibronectin, types I and III collagen in the acupotomy group were obviously decreased (all P<0.05). Compared with the acupotomy group, ROM, paw area, Max dA/dT, and MWW were restored (all P<0.05), and mRNA levels of fibrosis-related genes along with protein levels of Wnt 1, ß-catenin, fibronectin, types I and III collagen in the acupotomy 3-w group were decreased (P<0.05). CONCLUSION: Improvements in motor function, muscle contractures, and muscle fibrosis induced by acupotomy correlates with the inhibition of Wnt/ß-catenin signaling pathway.

[Effect of acupotomy on expressions of cellular senescence markers p16Ink4a and p21Waf1/Cip1 in cartilage tissue of rabbits with knee osteoarthritis].

OBJECTIVE: To observe the effect of acupotomy on the expressions of p16Ink4a and p21Waf1/Cip1 in knee osteoarthritis (KOA) rabbits，so as to analyze whether acupotomy can treat KOA by inhibiting the cellular senescence of chondrocytes. METHODS: Twenty-four New Zealand male rabbits were randomly divided into normal, model, acupotomy and electroacupuncture (EA) groups, with 6 rabbits in each group. The KOA model was established by left hindlimb straightening fixation. After modeling, rabbits in the acupotomy group were treated with acupotomy loosening therapy on high stress points around the affected knee joints such as tendons attachment points of vastus medialis, vastus lateralis, rectus femoris, biceps femoris and pes anserine bursa, once a week for 3 weeks. In the EA group, "Xuehai"(SP10), "Liangqiu" (ST34),"Neixiyan" (EX-LE4) and "Waixiyan" (ST35) on the affected hindlimb were selected for EA treatment (3 mA, 2 Hz/100 Hz), 20 min each time, once every other day for 3 weeks. Before and after treatments, the knee Lequesne MG score and passive range of motion (PROM) of the affected knee joint were evaluated. After the treatments, the expressions of p16Ink4a and p21Waf1/Cip1 in the cartilage tissue of the affected knee joint were detected by immunohistochemistry and Western blot respectively. RESULTS: Before and after treatment, compared with the normal group, the Lequesne MG score was significantly increased (P<0.01), the PROM was significantly decreased (P<0.01) in the model group. After treatment, compared with the normal group, the positive expression and protein expression levels of p16Ink4a and p21Waf1/Cip1 were significantly increased (P<0.01) in the model group; compared with the model group, the Lequesne MG score was significantly decreased (P<0.01), the PROM was significantly increased (P<0.01), the positive expression and protein expression levels of p16Ink4a and p21Waf1/Cip1 were significantly decreased (P<0.01，P<0.05) in the acupo-tomy and EA groups; compared with the EA group, the Lequesne MG score was decreased (P<0.05), the PROM was increased (P<0.05), the positive expression and protein expression levels of p16Ink4a and p21Waf1/Cip1 were decreased (P<0.05，P<0.01) in the acupotomy group. CONCLUSION: Acupotomy intervention can down-regulate the expressions of cellular senescence markers p16Ink4a and p21Waf1/Cip1 in chondrocytes, indicating that acupotomy therapy may alleviate cartilage degeneration by inhibiting chondrocyte premature cellular senescence to treat KOA.

Versatile Au@Ru nanocomposites for the rapid detection of Salmonella typhimurium and photothermal sterilization.

In view of the current public health hazards of food-borne pathogens, it is urgent to develop a rapid detection method with high sensitivity, good specificity and operational convenience, as well as to determine an effective sterilization strategy. Herein, versatile gold-ruthenium nanocomposites modified with antibody (Au@Ru-pAb Ncs) have been constructed for the sensitive detection of Salmonella typhimurium (S. typhimurium) via the lateral flow immunochromatographic assay (LFIA). Au@Ru-pAb Ncs based LFIA exhibited a wide detection range from 2.9 × 106 CFU/mL to 2.9 × 1011 CFU/mL with the limit of detection of 9.8 × 104 CFU/mL for S. typhimurium, and displayed excellent specificity. In addition, Au@Ru-pAb Ncs irradiated with 808 nm (500 mW/cm2) near-infrared light (NIR) had a significant antibacterial effect within only 5 min, attributed to its high photothermal conversion efficiency of 54.14%. Therefore, both sensitive detection of S. typhimurium and effective NIR-triggered photothermal sterilization were achieved by using versatile Au@Ru-pAb Ncs, showing great prospects in the field of pathogen detection and treatment.

An ultrasensitive colorimetric assay based on a multi-amplification strategy employing Pt/IrO2@SA@HRP nanoflowers for the detection of progesterone in saliva samples.

Progesterone (P4) belongs to a factor that affects stress response and is a potential carcinogen, and saliva levels are expected to be a standard measurement for clinical diagnosis. In this study, a new type of nanoflower with both recognition functionality and catalytic substrate ability was prepared by copper phosphate, Pt/IrO2 nanocomposites (Pt/IrO2 NPs), streptavidin (SA) and horseradish peroxidase (HRP) via a one-pot co-precipitation strategy. Due to the enhanced catalytic activity and stability of Pt/IrO2@SA@HRP nanoflowers, we developed a powerful and sensitive multiple-catalysis ELISA to monitor progesterone in saliva. Multiple-catalysis ELISA based on a specific antibody and Pt/IrO2@SA@HRP nanoflowers exhibited a linear interval range from 0.217 ng mL-1 to 7.934 ng mL-1. The median inhibitory concentration (IC50) for progesterone is 1.311 ng mL-1 and the limit of detection (LOD = IC10) is 0.076 ng mL-1 in the proposed method. Satisfactory recoveries were in a range of 79.6-107% with an acceptable coefficient of variation (below 10.6%). Results of the multiple-catalysis ELISA and LC-MS/MS had a good coincidence. Our result unraveled that multiple-catalysis ELISA is a potentially serviceable tool for the detection of progesterone in saliva.

Lateral Flow Immunosensor for Ferritin Based on Dual Signal-Amplified Strategy by Rhodium Nanoparticles.

Lateral flow immunoassay (LFIA) is one of the most widely used tools for analysis and field measurement and has the advantages of high efficiency, simple operation, portability, and low cost. Therefore, in this study, we designed a proof-of-concept of LFIA based on rhodium nanoparticles and investigated its improvement by further introducing the tetramethyl benzidine and H2O2 mixture as the substrate to trigger the color reaction. The proposed methods were qualitative research by the naked eye and quantitative measurement by a smartphone and software. Under the optimal condition, the detection of ferritin was successfully established with the limit of detection of 0.3 ng/mL. The lowest visually detectable amount was 0.05 ng/mL. To verify the performance of the RhNPs-LFIA, three spiked serum samples were tested, and the recovery rate increased from 88.9 to 129.9%, revealing that the proposed methods were applicable and practically reliable for testing serum samples. The developed RhNP-based LFIA is highly sensitive and convenient, which provides a promising technology for accurate, rapid, high sensitivity, and high screening detection of ferritin in clinical diagnosis.