A clinician's perspective on boron neutron capture therapy: promising advances, ongoing trials, and future outlook.

PURPOSE: This comprehensive review aims to provide a unique clinical perspective on the latest advances and ongoing boron neutron capture therapy (BNCT) trials for various cancers. METHODS: We critically analyzed clinical data from BNCT trials for head and neck cancer, glioblastoma, melanoma, meningioma, breast cancer, and liver tumors. We investigated differences in tumor responses and normal tissue toxicities among trials and discussed potential contributing factors. We also identified the limitations of early BNCT trials and proposed strategies to optimize future trial design. RESULTS: BNCT has shown promising results in treating head and neck cancer, with high response rates and improved survival in patients with recurrent disease. In glioblastoma, BNCT combined with surgery and chemotherapy has demonstrated survival benefits compared to standard treatments. BNCT has also been successfully used for recurrent high-grade meningiomas and shows potential for melanomas, extramammary Paget's disease, and liver tumors. However, differences in tumor responses and toxicities were observed among trials, potentially attributable to variations in treatment protocols, patient characteristics, and evaluation methods. CONCLUSIONS: BNCT is a promising targeted radiotherapy for various cancers. Further optimization and well-designed randomized controlled trials are needed to establish its efficacy and safety. Future studies should focus on standardizing treatment protocols and addressing limitations to guide clinical decision-making and research priorities.

Effect of Stewing Time on the Small Molecular Metabolites, Free Fatty Acids, and Volatile Flavor Compounds in Chicken Broth.

Chicken broth has a taste of umami, and the stewing time has an important effect on the quality of chicken broth, but there are fewer studies on the control of the stewing time. Based on this, the study was conducted to analyze the effects of different stewing times on the sensory, small molecular metabolites, free fatty acids, and volatile flavor compounds contents in chicken broths by liquid chromatography-quadrupole/time-of-flight mass spectrometry, gas chromatography-mass spectrometry, headspace solid-phase microextraction, and gas chromatography-mass spectrometry. Eighty-nine small molecular metabolites, 15 free fatty acids, and 86 volatile flavor compounds were detected. Palmitic and stearic acids were the more abundant fatty acids, and aldehydes were the main volatile flavor compounds. The study found that chicken broth had the best sensory evaluation, the highest content of taste components, and the richest content of volatile flavor components when the stewing time was 2.5 h. This study investigated the effect of stewing time on the quality of chicken broth to provide scientific and theoretical guidance for developing and utilizing local chicken.

Effect of different salt additions on the taste and flavor-related compounds in chicken soup.

Chicken soup is popular among consumers because of its delicious taste, strong flavor, and abundant nutritional value. Twenty-four Yunnan local hens were stewed by adding different amounts of NaCl [1.5, 2, 2.5, 3%, m/m, calculated based on chicken carcass weight; chicken: water = 1:2 (m/m)] to study the effect of salt addition on taste- and flavor-related compounds in chicken soup. Sensory evaluation results showed that the 2 and 2.5% NaCl treatment groups had higher scores. Water-soluble small molecule compounds were detected by LC-Q/TOF-MS based metabolomics approach, among which amino acids and their derivatives, nucleic acids, and small peptides were the main components. The concentration of Water-soluble small molecule substances in chicken soup samples with different salt additions showed a clear trend of separation and reached the highest in the 2.5% NaCl treatment group. Volatile flavor compounds in the chicken soup were analyzed by HS-SPME-GC-MS, including aldehydes, and alcohols, and the relative concentration of flavor compounds in the 2.5% salt treatment group was the highest. In summary, the addition of salt could improve the overall flavor of chicken broth, and the optimal salt addition of NaCl in chicken soup is 2.5%.

Precise Molecular Profiling of Circulating Exosomes Using a Metal-Organic Framework-Based Sensing Interface and an Enzyme-Based Electrochemical Logic Platform.

Exosomes have emerged as a promising circulating tumor biomarker; however, it is a big challenge for convenient, multiparametric, and accurate profiling of tumorous exosomes due to their unique structure and heterogeneity. To address these problems, we develop a highly integrated electrochemical platform for molecular profiling of tumor exosomes. A metal-organic framework-functionalized sensing interface is fabricated through a simple self-growth process, which collects exosomes from biofluids without additional separation steps. Meanwhile, a sensing strategy is designed to analyze both exosomal protein and RNA markers on a single chip based on the unique sensor architecture, allowing detection of low-abundance targets (∼250 vesicles in a 10 µL sample) using an integrated microfluidic electrochemical device. Furthermore, a multiple-input, protein enzyme-based logic gate is introduced into the system to accurately identify breast cancer patients with 100% sensitivity and specificity, thus revealing the advantageous role of logical profiling of exosomes in early diagnostics of tumor.

Aptamer-Linked CRISPR/Cas12a-Based Immunoassay.

The clustered regularly interspaced short palindromic repeat (CRISPR)/Cas system has shown great promising applications in the area of nucleic acid biosensing. However, because of the dearth of versatile signal transduction strategies, this system is usually compromised to low versatility, moderate sensitivity, and complex operation for non-nucleic acid targets, limiting its clinical transition. Herein, we describe a direct method to establish the correlation between non-nucleic acid analytes and the CRISPR/Cas12a system using a series of rationally designed, aptamer-flanked activator DNA strands, which enable ultrasensitive detection of biomarkers from different species, greatly broadening the possibility of the CRISPR/Cas system in bioanalysis. Meanwhile, the signal output is highly optional and the sensing principle is akin to the traditional enzyme-linked immunosorbent assay (ELISA), so it can be directly imposed on the currently available ELISA platform, further facilitating its application in medical diagnostics.

Coating a DNA self-assembled monolayer with a metal organic framework-based exoskeleton for improved sensing performance.

The formation of DNA self-assembled monolayers (SAMs) is one of the most popular ways to attach DNA molecules onto Au surfaces and is extensively used in many fields, especially in biosensing. However, the relatively poor stability of DNA SAMs (e.g., after long-term storage or under harsh environmental conditions) greatly limits their use in real applications. Herein, a new strategy is reported to protect the DNA SAMs by using a metal organic framework (MOF)-based exoskeleton. Taking electrochemical DNA (E-DNA) sensors as an example, we have systematically studied the stability of various DNA probes from the simple single-stranded DNA to a complex DNA nanostructure on the Au electrode surface. We have found that different DNA probes lead to various MOF profiles and the formed MOFs can be conveniently removed by simple acidic water rinse. Thanks to the exoskeleton, the stability of DNA SAMs is significantly enhanced and the DNA probes can be insulated from heat, nuclease, and varying ionic strengths, greatly extending the shelf-life of E-DNA sensors and indirectly improving their sensing performance. More importantly, the secondary structures of DNA probes can also be well preserved. The longstanding stability is of particular importance to biosensors; thus they can be facilely handled, transported, and stored in a resource-limited setting without compromising the analytical performance of biosensors.

Bridging exosome and liposome through zirconium-phosphate coordination chemistry: a new method for exosome detection.

We have proposed a new exosomes-zirconium-liposomes sandwich structure to detect exosomes by using zirconium-phosphate coordination chemistry. The combined use of the intrinsic property of phosphate in both exosomes and liposomes as well as zirconium ions can endow the method lower cost, no modified label, simplicity and high efficiency.

A reusable electrochemical sensor for one-step biosensing in complex media using triplex-forming oligonucleotide coupled DNA nanostructure.

Here we report an electrochemical DNA (E-DNA) sensor to detect a variety of analytes by using a novel interfacial probe that rationally integrates triplex-forming oligonucleotide (TFO) into a tetrahedral DNA nanostructure (TDN). In the presence of analyte, the blocked TFO is released and subsequently binds the edge of TDN to form a triplex DNA structure, which confines the redox reporter to be in close proximity to the underlying electrode and enhances the electrochemical signal. Thanks to the unique design and property of the probe, the proposed sensor could efficiently suppress the background signal (from 0.69 µA to 0.092 µA) and thus enhance the signal-to-noise ratio, resulting in improved sensing performance. Furthermore, the sensor displays new merits such as rapid response (∼35 min), one-step operation, easy regeneration (buffer change) and good generality (changing recognition element) compared with traditional TDN-based E-DNA sensor using enzyme displays signal transducer. In addition, to demonstrate real-world applicability of this new sensor, we have successfully detected different analytes (e.g., DNA, protein, and metal ion) in the complex media (e.g., serum, blood, and lake water), implying its considerable potential for precise bioanalysis in the future.

A programmed terminal extension strategy to light up multiple beacons for DNA and cellular telomerase detection.

Here we develop a new method for the sensitive detection of DNA and cellular telomerase using an enzyme-triggered terminal extension strategy that the produced strand can light up multiple beacons on the surface of gold nanoparticles.

Supramolecular Chemistry-Assisted Electrochemical Method for the Assay of Endogenous Peptidylarginine Deiminases Activities.

Peptidylarginine deiminase 4 (PAD4) is the only isoform of PADs located within the cell nucleus, which has been known to be related to several human diseases. In this work, we have proposed an electrochemical method for the assay of endogenous PAD4 activities as well as the studies of PAD4 inhibitors by making use of the supramolecular chemistry-assisted signal labeling. Specifically, peptide probes P1 and P2, which separately contain cysteine residues and tripeptides FGG (Phe-Gly-Gly), can be self-assembled onto the surface of the gold electrode and silver nanoparticles, respectively. In the meantime, the peptide probes can be connected together through cucurbit[8]uril-mediated host-guest interaction. Nevertheless, after trypsin-catalyzed digestion, FGG at the N-terminal of P1 will be removed from the electrode surface, thereby inhibiting the connection of P1 and P2. Since PAD4 catalyzes the citrullination of arginine residue within P1, trypsin-catalyzed digestion of P1 can be prohibited by the addition of PAD4. Consequently, an obvious change of the electrochemical response can be obtained from the silver nanoparticles (AgNPs) immobilized on the electrode surface. Experimental results have shown that our method can display an improved sensitivity and specificity for both PAD4 assay and inhibitor screening, which may effectively trace endogenous PAD4 and the inhibitors in the cancer cells. Therefore, our method may have great potential for the diagnosis and treatment of PAD4-related diseases in the future.

Evaluating Tumor-Associated Activity of Extracellular Sulfatase by Analyzing Naturally Occurring Substrate in Tumor Microenvironment of Hepatocellular Carcinoma.

The progress of cancer is intimately connected with the activity of the extracellular matrix (ECM) enzymes. To evaluate the promoting effect of these enzymes on tumor development in a pathological biocontext, we propose in this work to analyze their natural substrates in the ECM. This strategy is demonstrated by studying heparan sulfate (HS), the substrate of ECM sulfatase, in the development of hepatocellular carcinoma (HCC). An assay is designed to study the abundance and sulfation of HS and to evaluate the interactions between HS and the growth factors, such as fibroblast growth factor 2 (FGF2). Peptides derived from the amyloid peptide and various growth factors are employed to detect HS and evaluate their affinity toward the growth factors, whereas the ruthenium polypyridyl complex is taken as a photocatalyst to achieve a more sensitive signal readout. Applying this method to HepG2 cells, correlated changes between the activity of sulfatase 2 in regulating FGF2-induced cell proliferation and the abundance, degree of sulfation, and growth factor binding of HS can be observed. This method has also been applied to analyze clinical tissue samples of HCC. The results may suggest tumor-progress-related alterations in the above-studied biochemical features of HS. These results may point to the prospect of using this method to facilitate the diagnosis and prognosis of HCC in the future.

Improvement of enzyme-linked immunosorbent assay for the multicolor detection of biomarkers.

An enzyme-linked immunosorbent assay that is dependent on enzyme amplification has dominated the current field of protein detection; however, limited multiple detection ability and susceptible enzymatic reactions, and low sensitivity may severely hinder its application. Here, we report a new signal amplification scheme based on allochroic molecule modified carboxyl graphene oxide (cGO), which can be used to develop a multicolor immunoassay named as allochroic-cGO linked immunosorbent assay (ALISA). Thanks to high adsorption levels and a wide selection of allochroic molecules, the simultaneous colorimetric detection of diagnostic biomarkers at a picogram level can be successfully achieved for the first time. In addition, the color change triggered by acidic or basic water can provide a simple, rapid, stable and economical signal output, further meeting the growing biodetection requirements. Moreover, with the help of ALISA, we demonstrate that the combined detection of three tumor biomarkers, including carcino-embryonic antigen, neuron-specific enolase, and cytokeratin-19 fragment, is more valuable for differentiating lung cancer patients than the detection of a single biomarker, further manifesting the superiority of ALISA. All in all, this straightforward approach not only opens up new prospects for multicolor immunoassays, but also has great potential for applications in resource-constrained settings.

An array-based approach to determine different subtype and differentiation of non-small cell lung cancer.

Simple and accurate methods of discriminating subtype or differentiation of human tumor are critical for designing treatment strategies and predicting disease prognosis, and the currently used method to determine the two important factors mainly depends on histological examination by microscopy observation, which is laborious, highly trained operator required, and prone to be disruptive due to individual-to-individual judgment. Here we report a novel array-based method based on the interaction of graphene oxide (GO) and single-strand DNA modified gold nanoparticles (ssDNA-AuNPs) to distinguish between different subtypes and grades of tumors through their overall intracellular proteome signatures. Strategically, we first select eight proteins at 0.5 nM concentration in buffer or 10 nM in human serum to verify the discriminant ability of our method, then choose adenocarcinoma and squamous-cell carcinoma that account for 90% non-small cell lung cancer, as well as their respective three tumor grades as model system to provide a realistic testing ground for clinical cancer analysis. Consequently, total differentiation between different subtype and grade of tumor tissues has been achieved with as little as 100 ng of intracellular protein, suggesting the high sensitivity and selectivity of this sensor array. Overall, this array-based approach may provide the possibility for unbiased and simplified personalized tumor classification diagnostics in the future.

MicroRNA-409-3p functions as a tumor suppressor in human lung adenocarcinoma by targeting c-Met.

BACKGROUND/AIMS: Dysregulation of microRNAs is correlated with tumor development. The aim of this study is to investigate the clinicopathologic and prognostic significance of microRNA (miR)-409-3p and its tumor suppressor roles in lung adenocarcinoma (LAD). METHODS: Quantitative real-time PCR (qRT-PCR) was performed to detect miR-409-3p expression in LAD tissues and corresponding noncancerous tissues. Additionally, the correlations of miR-409-3p expression with clinicopathologic factors and prognosis of patients were statistically analyzed. Next, we investigated whether miR-409-3p could function as a tumor suppressor in LAD cells via regulation of Akt signaling by targeting receptor tyrosine kinase (c-Met). RESULTS: MiR-409-3p was significantly downregulated in LAD tissues compared with corresponding noncancerous tissues. Low miR-409-3p expression was observed to be significantly correlated with poorer tumor differentiation, advanced pTNM stage and higher incidence of lymph node metastasis. Multivariate Cox regression analyses showed that miR-409-3p expression was an independent prognostic factor for LAD patients. Functional analyses indicated that miR-409-3p could inhibit growth, induce apoptosis, reduce migration and invasion in LAD cells via inactivation of Akt signaling by targeting c-Met. CONCLUSIONS: MiR-409-3p was an independent prognostic factor and functioned as a tumor suppressor in LAD via regulation of Akt signaling by targeting c-Met.

Enhanced charge transfer by gold nanoparticle at DNA modified electrode and its application to label-free DNA detection.

Rational utilization of nanomaterials to construct electrochemical nucleic acid sensors has attracted large attention in recent years. In this work, we systematically interrogate the interaction between gold nanoparticles (GNPs) and single-strand DNA (ssDNA) immobilized on an electrode surface and then take advantage of the ultrahigh charge-transfer efficiency of GNPs to develop a novel DNA sensing method. Specifically, ssDNA modified gold electrode can adsorb GNPs because of the interaction between gold and nitrogen-containing bases; thus, the negative electrochemical species [Fe(CN)6](3-/4-) may transfer electrons to electrode through adsorbed GNPs. In the presence of target DNA, the formed double-strand DNA (dsDNA) cannot capture GNPs onto the electrode surface and the dsDNA may result in a large charge-transfer resistance owing to the negatively charged phosphate backbones of DNA. So a simple but sensitive method for the detection of target DNA can be developed by using GNPs without any requirement of modification. Experimental results demonstrate that the electrochemical method we have proposed in this work can detect as low as 1 pM breast cancer gene BRCA1 in a 10 µL sample volume without any signal amplification process or the involvement of other synthesized complex, which may provide an alternative for cancer DNA detection. This method may also be generalized for detecting a spectrum of targets using functional DNA (aptamer, metal-specific oligonucleotide, or DNAzyme) in the future.

Lignin binding to pancreatic lipase and its influence on enzymatic activity.

In this paper, we find that the effect of lignin on pancreatic lipase (PL) is dependent on reaction medium and substrate used. Experimental results reveal that lignin can gradually bind to PL to form a PL-lignin complex, resulting in an increased activity of the enzyme. The binding process is spontaneous and the PL-lignin complex formation is an endothermic reaction induced by hydrophobic and electrostatic interaction. There is a non-radiation energy transfer from PL to lignin during the binding process, and the binding of lignin to PL conforms to a secondary exponential decay function. Moreover, the α-helix content of the enzyme will be changed and the rigidity of its side chain will be enhanced due to the formation of lignin-PL complex. This study has not only provided the activation effect of lignin on PL, but also given an insight into the interaction between lignin and the enzyme, which would benefit the application of lignin in the pharmacy and food industry, as well as other fields.