1796 Introduction: The Polymerase Chain Reaction (PCR) has become the cornerstone of molecular diagnosis, with almost every genetic test aiming to identify DNA sequence variation incorporating PCR. A commonly encountered problem is that variant DNA sequences exist in the presence of a large majority of wild type alleles such as when DNA is obtained from heterogeneous cancer biopsies or when fetal alleles are sought in maternal blood. As a result, downstream assays are severely limited in their ability to identify subtle genetic changes that can have profound impact in clinical decision-making and outcome. We describe Co-amplification at Lower Denaturation temperature (COLD-PCR), a novel form of PCR that amplifies minority alleles selectively from mixtures of wild-type and mutation-containing sequences irrespective of the mutation type or position on the sequence (Nature Medicine, In Press).  Methods: In COLD-PCR, an intermediate annealing temperature is used during PCR-cycling to allow cross-hybridization of mutant and wild type alleles; hetero-duplexes, which melt at lower temperatures than homo-duplexes, are then selectively denatured and amplified at Critical Denaturation Temperature (Tc), while homo-duplexes remain double-stranded and do not amplify efficiently. By fixing the denaturation temperature to Tc, mutations at any position along the sequence are enriched during COLD-PCR amplification. To validate COLD-PCR, we used serial dilutions of DNA from tumor-derived cell lines containing point mutations or micro-deletions at different positions along p53 exon 8 and Kras exon 1 (codons 12-13), or samples with micro-deletions in EGFR exon 19. In addition, genomic DNA from a series of colon and lung cancer surgical specimens, and plasma-circulating DNA collected under IRB approval were utilized for validation on clinical specimens.  Results: By replacing regular PCR with COLD-PCR prior to application of a range of assays (Sanger sequencing; Pyrosequencing; MALDI-TOF; dHPLC; RFLP; and Taqman) we improved mutation detection sensitivity up to 100-fold and identified several additional p53/Kras/EGFR mutations in heterogeneous cancer samples. 4 of 43 surgical samples and 3 of 10 plasma samples tested contained clinically important mutations that were not detected by any of the methods tested when preceded by regular-PCR, but they were detectable following COLD-PCR.  Conclusion: Replacement of regular PCR with COLD-PCR provides a universal boost to most mutation detection technologies and enables them to be used with the required confidence in routine screening of cancer specimens for somatic mutations, including low-level mutation screening of surgical/FFPE tumor samples or bodily fluids. COLD-PCR is also anticipated to find application in pre-natal diagnosis, in infectious diseases and in epigenetics.

Replacing PCR with COLD-PCR enriches variant DNA sequences and redefines the sensitivity of genetic testing in cancer

Targeted cancer therapy has become one of the most important medical methods because of the spreading and metastatic nature of cancer. Based on the introduction of AS1411 and its four-chain structure, this paper reviews the research progress in cancer detection and drug delivery systems by modifying AS1411 aptamers based on graphene, mesoporous silica, silver and gold. The application of AS1411 in cancer treatment and drug delivery and the use of AS1411 as a targeting agent for the detection of cancer markers such as nucleoli were summarized from three aspects of active targeting, passive targeting and targeted nucleic acid apharmers. Although AS1411 has been withdrawn from clinical trials, the research surrounding its structural optimization is still very popular. Further progress has been made in the modification of nanoparticles loaded with TCM extracts by AS1411. 

https://jnanobiotechnology.biomedcentral.com/counter/pdf/10.1186/s12951-022-01240-z

Progress in cancer drug delivery based on AS1411 oriented nanomaterials

Nanofibrous hydrogels embedded with phase-change materials: Temperature-responsive dressings for accelerating skin wound healing

Multifunctional nanoparticles as optical biosensing probe for breast cancer detection: A review

Polymer composites are ubiquitous and indispensable in numerous applications. Coupling the inclusion of reinforcing agents with methods developed to control filler distribution and orientation allow for significant enhancement in mechanical, thermal, electrical, and barrier/transport-related properties of composites. For practical implementation, ideal approaches to fabricating polymer composites with directionally assembled fillers must consider manufacturing compatibility, scale-up ease, and aligning efficiency. In this article, we will review a cost-effective and industrially relevant toolbox for preferential filler alignment in polymer composites. Three main strategies for aligning fillers within polymer composites will be discussed. Specifically, solution casting and compression molding can introduce compression force to assemble fillers along the in-plane direction of composite films, shear force can be developed during fiber spinning, film blowing, and uniaxial/biaxial stretching processes to align fillers along the shear direction, and external fields (both electric and magnetic fields) can direct assembly of fillers, typically along the out-of-plane direction. For each section, we not only highlight the mechanisms of these methods but also demonstrate the critical structure–property relationships through model examples. Finally, a perspective on future opportunities and challenges of anisotropic and performance-enhanced polymer composite preparation strategies is provided, with a particular focus on additive manufacturing. 

Scalable methods for directional assembly of fillers in polymer composites: Creating pathways for improving material properties

Nucleic acid based nanocomposites and their applications in biomedicine

The invention discloses a surface-enhanced Raman-fluorescent duel-mold nano-probe based on conjugated polymer and a preparation method of the duel-mold nano-probe. The probe is of a four-layer core-shell structure which sequentially comprises a noble metal nano-particle inner core, a first modifying layer, a conjugated polymer fluorescent molecule layer and a second modifying layer from inside tooutside, wherein the noble metal nano-particle inner core is linked with Raman signal molecules, the core is packaged by the first modifying layer, the first modifying layer is modified with the conjugated polymer fluorescent molecule layer, and the conjugated polymer fluorescent molecule layer is packaged with the second modifying layer. According to the nano-probe, conjugated polymer nano-particles are taken as a fluorescence signal source, so that the photochemical stability is improved, and the biotoxicity is reduced; and by controlling the thickness of the first modifying layer, the distance from a fluorescent molecule to a metal core is regulated and controlled, so that the optical property of the probe is regulated and controlled. The SERS-MEF duel-mold probe simultaneously has dualadvantages of a surface-enhanced Raman spectrum and metal-enhanced fluorescence, is capable of realizing multivariate analysis, rapid imaging and the like, can be widely applied to biomedicine detection and has relatively wide application prospects.

Surface-enhanced Raman-fluorescent duel-mold nano-probe based on conjugated polymer and preparation method of duel-mold nano-probe

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Nucleic acid based nanocomposites and their applications in biomedicine

Surface-enhanced Raman-fluorescent duel-mold nano-probe based on conjugated polymer and preparation method of duel-mold nano-probe