Dual amplification dynamic DNA network system for CRISPR/Cas12a based p53 gene detection.

BACKGROUND: It is estimated that over 50 % of human cancers are caused by mutations in the p53 gene. Early sensitive and accurate detection of the p53 gene is important for diagnosis of cancers in the early stage. However, conventional detection techniques often suffer from strict reaction conditions, or unsatisfied sensitivity, so we need to develop a new strategy for accurate detection of p53 gene with smart designability, multiple signal amplification in mild reaction conditions. RESULTS: In this study, CRISPR/Cas system is exploited in entropy-driven catalysis (EDC) and hybridization chain reaction (CHA) dual signal amplification sensing strategies. The products of both reactions can efficiently and separately activate CRISPR/Cas12a which greatly amplifies the fluorescent signal. The method has good linearity in p53 detection with the concentration ranged from 0.1 fM to 0.5 pM with ultra-low detection limit of 0.096 fM. It also showed good performance in serum, offering potentials for early disease detection. SIGNIFICANCE: The designed dual amplification dynamic DNA network system exhibits an ultra-sensitive fluorescence biosensing for p53 gene identification. The method is simple to operate and requires only one buffer for the experiment, and meanwhile shows smart designability which could be used for a wide range of markers. Thus, we believe the present work will provide a potential tool for the construction and development of sensitive fluorescent biosensors for diseases.

An integrated bipolar electrode with shared cathode for dual-mode detection and imaging of CEA.

In this paper, we designed a novel shared cathode bipolar electrode chip based on Ohm 's law and successfully constructed a dual-mode dual-signal biosensor platform (DD-cBPE). The device integrates ELISA, ECL, and ECL imaging to achieve highly sensitive detection and visual imaging of carcinoembryonic antigen (CEA). The unique circuit structure of the device not only realizes the dual signal detection of the target, but also breaks the traditional signal amplification concept. The total resistance of the system is reduced by series-parallel connection of BPE, and the total current in the circuit is increased. In addition, Au@NiCo2O4@MnO2 nanozyme activity probe was introduced into the common cathode to enhance the conductivity of the material. At the same time, due to the excellent peroxidase (POD) activity of NiCo2O4@MnO2, the decomposition of H2O2 was accelerated, so that more electrons flowed to the BPE anode, and finally the dual amplification of the ECL signal was realized. The device affects the current in the circuit by regulating the concentration of the co-reactant TPrA, thereby affecting the resistance of the system. Finally, different luminescent reagents emit light at the same potential and the luminous efficiency is similar. In addition, the chip does not need external resistance regulation, which improves the sensitivity of the immunosensor and meets the needs of timely detection. It provides a new idea for the deviceization of bipolar electrodes and has broad application prospects in biosensors, clinical detection, and environmental monitoring.

Injectable and Thermosensitive Hydrogel with Platelet-Rich Plasma for Enhanced Biotherapy of Skin Wound Healing.

The rapid and effective healing of skin wounds resulted from severe injuries and full-layer skin defects remains a pressing clinical challenge in contemporary medical practice. The reduction of wound infection and rapid healing is helpful to rebuild and repair skin tissue. Here, a thermosensitive chitosan-based wound dressing hydrogel incorporating ß-glycerophosphate (GP), hydroxy propyl cellulose (HPC), graphene oxide (GO), and platelet-rich plasma (PRP) is developed, which exhibits the dual functions of antibacterial properties and repair promotion. GP and HPC enhance the mechanical properties through forming hydrogen bonding connection, while GO produces local heat under near-infrared light, leading to improved blood circulation and skin recovery. Notably, antibacterial properties against Pseudomonas aeruginosa, and control-release of growth factors from PRP are also achieved based on the system. In vitro experiments reveal its biocompatibility, and ability to promote cell proliferation and migration. Animal experiments demonstrate that the epithelial repair and collagen deposition can be promoted during skin wound healing in Sprague Dawley rats. Moreover, a reduction in wound inflammation levels and the improvement of wound microenvironment are observed, collectively fostering effective wound healing. Therefore, the composite hydrogel system incorporated with GO and PRP can be a promising dressing for the treatment of skin wounds.

Microperoxidase-11 functionalized nanozyme with enhanced peroxidase-mimicking activities for visual detection of cysteine.

Various nanomaterials with peroxidase activity (nanozyme) have been designed for bio catalysis and biosensing, however, most of them need further design and modification of probe molecules for the specific binding reaction with targets. This results in a decrease in catalysis activity and hinders them to be perfect alternatives to natural enzyme in biosensing. In this work, an enhanced nanozyme was synthesized by functionalizing natural microperoxidase-11 (MP-11) on a hybrid graphene oxide-gold (GO-Au) material. The designed nanozyme showed an enhanced catalysis activity and realized a robust and efficient colorimetric detection of cysteine based on specific binding reaction between active iron center from MP-11 and thiol in cysteine. The enhanced properties show promising applications of complex nanozyme and provides a great opportunity for developing efficient sensing systems.

Recent advances in nanotechnology-enhanced biosensors for α-fetoprotein detection.

α-Fetoprotein (AFP) is a kind of fetal protein that is related to tumor, the increasing concentration of which gives birth to a large variety of diseases, such as liver cancer. Therefore, the detection method with super sensitivity, high selectivity, and less time consumption under trace concentrations in early stage of diseases is becoming a necessity. In recent years, nanomaterials have been regarded as significant resources for the exploration of efficient biosensors with high sensitivity, selectivity, speed, as well as simple process, due to their excellent optical, electrical, and chemical properties. In this paper, we reviewed the research progress of AFP biosensors with enhanced sensitivity and selectivity by nanoparticles. Representative examples have also been displayed in this paper to expound the nanotechnologies utilized in the early detection of AFP. Furthermore, challenges of the clinical application of AFP biosensors based on nanotechnology have been elaborated, as well as the development opportunity in this field in the future. This review provides a comprehensive overview on the various nano-biosensor for AFP detection based on functional nanotechnology.

Platelet-rich plasma for the treatment of chronic rectal ulcer: A case report.

RATIONALE: Rectum ulcer is a disease of the digestive system, the main symptoms of which includes bloody diarrhea, abdominal pain, hematochezia, etc. At present, drug therapy and surgery are the most common treatments. Platelet-rich plasma (PRP) contains high concentrations of platelets and has been used to promote wound healing. However, the utilization of PRP in rectal ulcers has rarely been reported. PATIENT CONCERNS: The patient had reported a complaint of blood dripping from the stool for more than 4 months. She had a history of surgery for rectal cancer with postoperative chemotherapy and radiotherapy 19 years prior. Mesalazine suppository was given to her for about 4 months, and glutamine capsules for 2 months, but the rectal ulcer remained unhealed. DIAGNOSIS: A rectal ulcer was observed on colonoscopy, and the biopsy result was tubular adenoma. INTERVENTIONS: Autologous PRP treatment was performed for the patient under an anorectal scope together with basic supportive care. OUTCOMES: The ulcer nearly healed within 9 days after twice PRP treatments. LESSONS: PRP treatment may bring about novel treatment options for rectal ulcers.

Hemin-graphene oxide-gold nanoflower-assisted enhanced electrochemiluminescence immunosensor for determination of prostate-specific antigen.

An ultrasensitive luminol electrochemiluminescence (ECL) immunosensor was constructed for the detection of prostate specific antigen (PSA) using glucose oxidase-decorated hemin-graphene oxide-gold nanoflowers ternary nanocomposites as probes. Graphene oxide was first modified with hemin and then with gold nanoflowers through an in situ growth method, which has significantly boosted the catalytic activity of this graphene oxide-based peroxidase mimetics. The biocatalytical activity of this ECL immunosensor was thoroughly investigated to achieve selective recognition of the analyte molecules (PSA) by specific binding between antigens and antibodies. The limit of detection was calculated to be 0.32 pg mL-1 with a signal-to-noise ratio of 3. A broad linear range from 7.5 × 10-4 to 2.5 ng mL-1 was obtained. Such step-by-step assembled biosensor showed controlled nanostructure and exhibited promising application in analysis of human serum samples with a recovery range of 90.6-111.8% and a RSD range of 3.9-5.5%.