Cervical lymph node metastasis prediction from papillary thyroid carcinoma US videos: a prospective multicenter study.

BACKGROUND: Prediction of lymph node metastasis (LNM) is critical for individualized management of papillary thyroid carcinoma (PTC) patients to avoid unnecessary overtreatment as well as undesired under-treatment. Artificial intelligence (AI) trained by thyroid ultrasound (US) may improve prediction performance. METHODS: From September 2017 to December 2018, patients with suspicious PTC from the first medical center of the Chinese PLA general hospital were retrospectively enrolled to pre-train the multi-scale, multi-frame, and dual-direction deep learning (MMD-DL) model. From January 2019 to July 2021, PTC patients from four different centers were prospectively enrolled to fine-tune and independently validate MMD-DL. Its diagnostic performance and auxiliary effect on radiologists were analyzed in terms of receiver operating characteristic (ROC) curves, areas under the ROC curve (AUC), accuracy, sensitivity, and specificity. RESULTS: In total, 488 PTC patients were enrolled in the pre-training cohort, and 218 PTC patients were included for model fine-tuning (n = 109), internal test (n = 39), and external validation (n = 70). Diagnostic performances of MMD-DL achieved AUCs of 0.85 (95% CI: 0.73, 0.97) and 0.81 (95% CI: 0.73, 0.89) in the test and validation cohorts, respectively, and US radiologists significantly improved their average diagnostic accuracy (57% vs. 60%, P = 0.001) and sensitivity (62% vs. 65%, P < 0.001) by using the AI model for assistance. CONCLUSIONS: The AI model using US videos can provide accurate and reproducible prediction of cervical lymph node metastasis in papillary thyroid carcinoma patients preoperatively, and it can be used as an effective assisting tool to improve diagnostic performance of US radiologists. TRIAL REGISTRATION: We registered on the Chinese Clinical Trial Registry website with the number ChiCTR1900025592.

A triazine-based covalent organic framework decorated with cadmium sulfide for efficient photocatalytic hydrogen evolution from water.

Construction of inorganic/organic heterostructures has been proven to be a very promising strategy to design highly efficient photocatalysts for solar driven hydrogen evolution from water. Herein, we report the preparation of a direct Z-scheme heterojunction photocatalyst by in situ growth of cadmium sulfide on a triazine-based covalent organic framework (COF). The triazine based-COF was synthesized by condensation reaction of precursors 1,3,5-tris-(4-formyl-phenyl) triazine (TFPT) and 2,5-bis-(3-hydroxypropoxy) terephthalohydrazide (DHTH), termed as TFPT-DHTH-COF. Widely distributed nitrogen atoms throughout TFPT-DHTH-COF skeletons serve as anchoring sites for strong interfacial interactions with CdS. The CdS/TFPT-DHTH-COF composite showed a hydrogen evolution rate of 15.75 mmol h-1 g-1, which is about 75 times higher than that of TFPT-DHTH-COF (0.21 mmol h-1 g-1) and 3.4 times higher than that of CdS (4.57 mmol h-1 g-1). With the properly staggered band alignment and strong interfacial interaction between TFPT-DHTH-COF and CdS, a Z-scheme charge transfer pathway is achieved. The mechanism has been systematically analyzed by steady state and time-resolved photoluminescence measurements as well as in situ irradiated X-ray photoelectron spectroscopy.

Mesoporous Silica-Encapsulated Gold Nanorods for Drug Delivery/Release and Two-Photon Excitation Fluorescence Imaging to Guide Synergistic Phototherapy and Chemotherapy.

Photothermal therapy is a promising light-based medical treatment that relies on light absorption agents converting light irradiation into localized heat to destroy cancer cells or other diseased tissues. It is critical to enhance the therapeutic effects of cancer cell ablation for their practical applications. This study reports a high-performance combinational therapy for ablating cancer cells, including both photothermal therapy and chemotherapy to improve therapeutic efficiency. The prepared AuNR@mSiO2 loading molecular Doxorubicin (Dox) assemblies were highlighted by merits of facile acquisition, great stability, easy endocytosis, and rapid drug release in addition to improved anticancer capability upon irradiation with a femtosecond pulsed near-infrared (NIR) laser, where AuNR@mSiO2 nanoparticles afforded a high photothermal conversion efficiency of 31.7%. Two-photon excitation fluorescence imaging was introduced into confocal laser scanning microscope multichannel imaging to track the drug location and cell position in real time for monitoring the process of drug delivery in killing human cervical cancer HeLa cells and then to realize imaging-guiding cancer treatment. These nanoparticles exhibit widespread potential in photoresponsive utilizations including photothermal therapy, chemotherapy, one- and two-photon excited fluorescence imaging, and 3D fluorescence imaging and cancer treatment.

Investigation of Targeting Relationship between Micro-Rna-22 and Vegfr3 in Lung Squamous Cell Carcinoma.

AIM: The present study explored the clinical significance of microRNA-22 (miR-22) expression in lung squamous cell carcinoma and to explore the targeting relationship with vascular endothelial growth factor receptor 3 (VEGFR3). METHODS: A total of 49 patients with lung squamous cell carcinoma who underwent surgical treatment were selected. The expression of miR-22 was detected by fluorescence quantitative realtime PCR (qPCR), the expression of VEGFR3 was detected by Western blotting assays, and D240 labeled microlymphatic vessels density (MLVD) was detected by immunohistochemistry (IHC). Lung squamous cell carcinoma cell line SK-MES-1 was selected and the targeting relationship between miR-22 and VEGFR3 was analyzed by double luciferase reporter gene assay. Western blotting assays were used to detect the expression of vascular endothelial growth factor-D (VEGFD) and D240 in the blank control group, empty vector transfection group, miR-22 transfection group, miR-22 and VEGFR3 co-transfection group. RESULTS: The expression range of miR-22 in lung squamous cell carcinoma was 0.8-3.5. The expression of miR-22 in lung squamous cell carcinoma was significantly different by tumor maximum diameter, lymph node metastasis, vascular invasion and TNM stage. The expression of miR-22 was linked to survival time. There was a negative correlation between miR-22 and VEGFR3, miR-22 and MLVD. Double luciferase reporter gene assays showed that miR-22 reduced the luciferase activity of pGL3-VEGFR3-WT transfected cells. Compared with the control group, the expression of VEGF-D and D2-40 in the miR-22 transfection group was significantly decreased. However, VEGF-D and D240 in the miR-22 and VEGFR3 co-transfection group reversed the changes. CONCLUSION: We assumed that the abnormal expression of miR-22 in lung squamous cell carcinoma may be involved in the development and progression of lung squamous cell carcinoma. MiR-22 negatively regulated the target gene VEGFR3 to mediate lymphangiogenesis. The expression of miR-22 may also be linked to the prognosis of the disease.

Cell-of-Origin Subtyping of Diffuse Large B-Cell Lymphoma by Using a qPCR-based Gene Expression Assay on Formalin-Fixed Paraffin-Embedded Tissues.

The well-established cell-of-origin (COO) algorithm categorizes diffuse large B-cell lymphoma (DLBCL) into activated B-cell-like (ABC) and germinal center B-cell-like (GCB) subgroups through gene expression profiling. We aimed to develop and validate a qPCR-based gene expression assay to determine the COO subgroups of DLBCL with formalin-fixed paraffin-embedded (FFPE) tissue. We first established a DLBCL transcriptome database of 1,016 samples retrieved from three published datasets (GSE10846, GSE22470, and GSE31312). With this database, we identified a qPCR-based 32-gene expression signature (DLBCL-COO assay) that is significantly associated with the COO subgroups. The DLBCL-COO assay was further validated in a cohort of 160 Chinese DLBCL patients. Biopsy samples from DLBCL patients with paired FFPE and fresh frozen tissue were collected to assign COO subtypes based on the immunohistochemistry (IHC) algorithm (Han's algorithm), DLBCL-COO assay, and global gene expression profiling with RNA-seq. For 111 paired FFPE and fresh DLBCL samples, the concordance between the IHC, qPCR, and RNA-seq methods was 77.5% and 91.9%, respectively. The DLBCL-COO assay demonstrated a significantly superior concordance of COO determination with the "gold standard" RNA-seq compared with the IHC assignment with Han's algorithm (P = 0.005). Furthermore, the overall survival of GCB patients defined by the DLBCL-COO assay was significantly superior to that of ABC patients (P = 0.023). This effect was not seen when the tumors were classified by the IHC algorithm. The DLBCL-COO assay provides flexibility and accuracy in DLBCL subtype characterization. These findings demonstrated that the DLBCL-COO assay might serve as a useful tool for guiding prognostic and therapeutic options for DLBCL patients.

Gold nanorod enhanced conjugated polymer/photosensitizer composite nanoparticles for simultaneous two-photon excitation fluorescence imaging and photodynamic therapy.

Two-photon photodynamic therapy (2P-PDT) is a novel minimal invasive cancer treatment method with advantages of deep penetration and intrinsic three-dimensionally localized activation to precisely target cancerous tissues. However, the therapeutic efficacy of 2P-PDT is limited by small two-photon absorption cross sections of conventional organic photosensitizers. In addition, traditional photosensitizers generally exhibit weak emission and lack imaging modality. In this work, conjugated polymers and gold nanorods (Au NRs) were integrated to fabricate nano-sized photosensitizers to improve the performance of molecular photosensitizers for 2P-PDT. A molecular photosensitizer, tetraphenylporphyrin, was encapsulated into the conjugated polymer PFV to form conjugated polymer nanoparticles (CPNs), which were then covalently linked to silica coated Au NRs. In these integrated nanoparticles, the two-photon optical properties of tetraphenylporphyrin were first enhanced by fluorescence resonance energy transfer from PFV, then further enhanced by Au NRs through plasmon resonance. A silica shell was utilized as the spacer between Au NRs and CPNs to optimize the enhancement effects. Through the combined enhancement effects of energy transfer and plasmon resonance, two-photon excitation fluorescence and two-photon induced singlet oxygen generation of tetraphenylporphyrin were enhanced by up to 980- and 792-fold, respectively, at a silica spacer thickness of 9 nm. The application of these nanoparticles as photosensitizers for simultaneous two-photon imaging and 2P-PDT treatment have been demonstrated on HeLa cancer cells with high brightness and significantly enhanced cancer cell killing efficiency. These nanoparticles can act as promising nano-photosensitizers for 2P-PDT with simultaneous imaging modality.

Simultaneous Imaging and Selective Photothermal Therapy through Aptamer-Driven Au Nanosphere Clustering.

Gold (Au) nanoparticles display enhanced near-infrared (NIR) photothermal effects upon the formation of clusters. We studied the photothermal properties of Au nanosphere clusters on the single-particle level using photothermal heterodyne imaging (PTHI) microscopy to understand the enhancement mechanisms. NIR photothermal responses of Au nanoparticle clusters were found to significantly increase from monomers to trimers. The averaged PTHI signal intensity of Au nanosphere dimers and trimers is ∼10 and ∼25 times that of monomers. The NIR photothermal effect of clustered nanospheres strongly correlates with their longitudinal plasmon mode. Clustered Au nanospheres were demonstrated to exhibit dual-capability NIR photothermal imaging and therapy of human prostate cancer cells with high efficiency and selectivity. This strategy can be potentially utilized for simultaneous cancer imaging and therapy with 3D selectivity.

Red emitting conjugated polymer based nanophotosensitizers for selectively targeted two-photon excitation imaging guided photodynamic therapy.

Two-photon excitation (2PE) photodynamic therapy (PDT) is a non-invasive technique for the treatment of cancer. However, its clinical applications are limited by small two-photon absorption cross section values of conventional photosensitizers. Here we designed multifunctional conjugated polymer based nanoparticles consisting of a conjugated polymer, a photosensitizer and a red-emitting dye, which can realize simultaneous 2PE red emission imaging and 2PE-PDT activities. The working principle is based on a 2PE fluorescence resonance energy transfer strategy from the conjugated polymer to photosensitizing and imaging agents. In these nanoparticles (NPs), the conjugated polymer, PPBF, was chosen as a two-photon light-harvesting material while the photosensitizer (tetraphenylporphyrin, TPP) and the red-emitting dye (TPD) were chosen as energy acceptors. The 2PE emission of TPP and TPD was enhanced by up to ∼161 and ∼23 times, respectively. The 2PE-PDT activity of these NPs was significantly improved compared with those NPs without PPBF by up to ∼149 times. Further surface-functionalization with folic acid (FA) groups allows these nanoparticles to exhibit selective affinity toward KB cancer cells. These NPs could act as novel 2PE conjugated polymer based nanoparticles combined with the advantages of low dark cytotoxicity, selective targeting and imaging-guided 2PE-PDT activities.

Pyrrolopyrrole aza boron dipyrromethene based two-photon fluorescent probes for subcellular imaging.

A series of pyrrolopyrrole aza boron dipyrromethene derivatives have been designed and synthesised as two-photon fluorescent probes. The bisalkynyl analogues, having a donor-π-donor quadrupolar skeleton, show a red-shifted Q band at ca. 700 nm in toluene and a two-photon absorption (TPA) cross-section up to 2349 GM at 1040 nm. To enable these dyes to be used for subcellular imaging, a branched oligoethylene glycol unit has been introduced to enhance their hydrophilicity and biocompatibility, and a potential organelle-selective group, namely triphenylphosphonium or morpholine moiety has also been added with a view to targeting the mitochondria or lysosomes respectively. The resulting conjugates are essentially non-aggregated in deionised water, exhibiting an intense Q band at 668 nm and a TPA cross-section up to 384 GM at 1040 nm. These spectral features have been analysed using density functional theory calculations, which suggest that the TPA is mainly due to the S0→ S2 transition. In the subcellular imaging study, it has been found that the triphenylphosphonium-containing derivative can localise in the lysosomes of HeLa human cervical carcinoma cells, which may be a result of its positive charge and the negative log P value (-2.46, P = partition coefficient), while the morpholine-substituted analogue does not exhibit preferential subcellular localisation.

Photoinduced Nickel-Catalyzed Chemo- and Regioselective Hydroalkylation of Internal Alkynes with Ether and Amide α-Hetero C(sp3)-H Bonds.

A direct hydroalkylation of disubstituted alkynes with unfunctionalized ethers and amides was achieved in an atom-efficient and additive-free manner through the synergistic combination of photoredox and nickel catalysis. The protocol was effective with a wide range of internal alkynes, providing products in a highly selective fashion. Notably, the observed regioselectivity is complementary to conventional radical addition processes. Mechanistic investigations suggest that the photoexcited iridium catalyst facilitated the nickel activation via single-electron transfer.

Controlled preparation of Au/Ag/SnO2 core-shell nanoparticles using a photochemical method and applications in LSPR based sensing.

A photochemical method for the controlled preparation of core-shell Au/Ag/SnO2 nanorods (NRs) and nanospheres (NSs) has been developed based on photo-induced electron transfer processes in the plasmonic metal-semiconductor system. Au/AgNR/SnO2 and Au/AgNS/SnO2 were prepared by the UV irradiation of a mixture of mesoporous SnO2 coated AuNRs, or AuNSs, and AgNO3, in which AgNO3 was reduced by electrons transferred from the photo-excited mesoporous SnO2 (semiconductor) to the gold (metal). This method allows precise control over the composition and optical properties of the obtained nanoparticles. The LSPR refractive index sensitivity of the obtained Au/AgNR/SnO2 nanoparticles has been optimized to obtain a refractive index sensitivity of ∼442 nm RIU(-1). The optimized nanoparticles were subsequently chosen for the LSPR based sensing of glutathione (GSH) with the limit of detection of ∼7.5 × 10(-7) M. This photochemical method allows the controlled preparation of various Au/Ag/SnO2 nanoparticles to adjust their LSPR to suit various applications.

Development of targetable two-photon fluorescent probes to image hypochlorous Acid in mitochondria and lysosome in live cell and inflamed mouse model.

Hypochlorous acid (HOCl), as a highly potent oxidant, is well-known as a key "killer" for pathogens in the innate immune system. Recently, mounting evidence indicates that intracellular HOCl plays additional important roles in regulating inflammation and cellular apoptosis. However, the organelle(s) involved in the distribution of HOCl remain unknown, causing difficulty to fully exploit its biological functions in cellular signaling pathways and various diseases. One of the main reasons lies in the lack of effective chemical tools to directly detect HOCl at subcellular levels due to low concentration, strong oxidization, and short lifetime of HOCl. Herein, the first two-photon fluorescent HOCl probe (TP-HOCl 1) and its mitochondria- (MITO-TP) and lysosome- (LYSO-TP) targetable derivatives for imaging mitochondrial and lysosomal HOCl were reported. These probes exhibit fast response (within seconds), good selectivity, and high sensitivity (<20 nM) toward HOCl. In live cell experiments, both probes MITO-TP and LYSO-TP were successfully applied to detect intracellular HOCl in corresponding organelles. In particular, the two-photon imaging of MITO-TP and LYSO-TP in murine model shows that higher amount of HOCl can be detected in both lysosome and mitochondria of macrophage cells during inflammation condition. Thus, these probes could not only help clarify the distribution of subcellular HOCl, but also serve as excellent tools to exploit and elucidate functions of HOCl at subcellular levels.

Red-emitting DPSB-based conjugated polymer nanoparticles with high two-photon brightness for cell membrane imaging.

New red-emitting conjugated polymers have been successfully synthesized by incorporating classical two-photon absorption (TPA) units, electron-rich units, and a small amount of electron-deficient units along the polymer backbones. Water-dispersible nanoparticles (NPs) based on these polymers were also fabricated for applications in two-photon excitation fluorescence imaging of cell membrane. Through optimization of the polymer/matrix mass ratio and the initial feed concentration of the polymer solution, a high quantum yield (QY) of 24% was achieved for the red-emitting NPs in water. TPA cross section and two-photon action cross section values of these polymers at 750 nm reached up to 1000 GM and 190 GM per repeat unit in aqueous media, 2.5 × 10(5) GM and 4.7 × 10(4) GM per NP, respectively. Furthermore, these NPs displayed excellent photostability and biocompatibility. Their applications as two-photon excitation fluorescence probes for cell membrane imaging have been demonstrated in three different cell lines with excellent imaging contrast. These results demonstrated that these polymer NPs hold great potentials as excellent two-photon excitation fluorescence probes in various biological applications.

Circulating CUDR, LSINCT-5 and PTENP1 long noncoding RNAs in sera distinguish patients with gastric cancer from healthy controls.

The examination of circulating nucleic acids (CNAs) is an emerging noninvasive diagnostic technique. However, it is unclear if serum long noncoding RNAs (lncRNAs) represent a novel marker to detect gastric cancer (GC). In this study, we measured 39 candidate cancer-associated lncRNAs by reverse transcription and quantitative polymerase chain reaction (RT-qPCR) in sera from 110 patients with GC, 106 age- and sex-matched healthy subjects and 15 patients with gastric peptic ulcer, markers were validated and assessed by RT-qPCR. The correlation of the expression levels of the candidate serum lncRNAs with clinical parameters of GC patients was performed. A three-lncRNA signature, including CUDR, LSINCT-5 and PTENP1, was identified that may be potential diagnostic marker for GC. The areas under the receiver operating characteristic (ROC) curve for this serum three-lncRNA signature were 0.920 and 0.829 for the two sets of serum samples. Moreover, a risk model for the serum three-lncRNA signature demonstrated that healthy samples can be distinguished from early GC samples. Three-lncRNA signature in serum was identified as diagnostic marker for GC. This work may facilitate the detection of GC and serve as the basis for further studies of the clinical value of serum lncRNAs in maintaining surveillance and forecasting prognosis.

Highly efficient, conjugated-polymer-based nano-photosensitizers for selectively targeted two-photon photodynamic therapy and imaging of cancer cells.

Two-photon photodynamic therapy (2P-PDT) is a promising noninvasive treatment of cancers and other diseases with three-dimensional selectivity and deep penetration. However, clinical applications of 2P-PDT are limited by small two-photon absorption (TPA) cross sections of traditional photosensitizers. The development of folate receptor targeted nano-photosensitizers based on conjugated polymers is described. In these nano-photosensitizers, poly{9,9-bis[6''-(bromohexyl)fluorene-2,7-ylenevinylene]-co-alt-1,4-(2,5-dicyanophenylene)}, which is a conjugated polymer with a large TPA cross section, acts as a two-photon light-harvesting material to significantly enhance the two-photon properties of the doped photosensitizer tetraphenylporphyrin (TPP) through energy transfer. These nanoparticles displayed up to 1020-fold enhancement in two-photon excitation emission and about 870-fold enhancement in the two-photon-induced singlet oxygen generation capability of TPP. Surface-functionalized folic acid groups make these nanoparticles highly selective in targeting and killing KB cancer cells over NIH/3T3 normal cells. The 2P-PDT activity of these nanoparticles was significantly improved, potentially up to about 1000 times, as implied by the enhancement factors of two-photon excitation emission and singlet oxygen generation. These nanoparticles could act as novel two-photon nano-photosensitizers with combined advantages of low dark cytotoxicity, targeted 2P-PDT with high selectivity, and simultaneous two-photon fluorescence imaging capability; these are all required for ideal two-photon photosensitizers.

In vitro, ex vivo and in vivo anti-hypertensive activity of Chrysophyllum cainito L. extract.

Chrysophyllum cainito L., a traditional herbal medicine, could have the potential for management of hypertension due to presence of polyphenolic compounds. The extracts and fractions of the pulp of plant were evaluated for in vitro (inhibition of angiotensin I converting enzyme/ACE assay), ex vivo (isolated aorta relaxation assay) and in vivo (salt induced hypertensive rat assay). The alcoholic and aqueous extract (ALE and AQE respectively) of fruit of plant C. cainito was having 14.8 and 9.2% yield respectively. The fractionation with ethyl alcohol (EAF) and butanol (BTF) yielded 2.52 & 2.17% respectively from ALE and 0.46 & 0.31% respectively from AQE with respect to fruit pulp dry weight. More phenolic content was found in ALE (3.75±0.15 mg gallic acid equivalent or GAE g(-1) of dry power of fruit pulp) compared to AQE and maximum in ethyl acetate fraction of ALE (ALE-EAF) (2.32±0.21 mg GAE g(-1) of dry power of fruit pulp) among all fractions. ACE inhibition activity was found to be maximum in ALE-EAF 62.5±7.34%. While ex vivo study using isolated tissue of aorta showed again showed maximum activity (62.82±6.19 and 46.47±8.32% relaxation with 50 µg mL(-1) and 10 µg mL(-1) GAE concentration respectively). ALE-EAF reduced the elevated arterial pressure of salt induced hypertensive rat significantly to the level of normotensive animal group. Results of ALE-EAF have shown its potential as a source for novel constituent for the treatment hypertension and should further be studied for isolation of specific constituent for more effectiveness.

Meta-analysis of associations between interleukin-17 gene polymorphisms and risk of gastric cancer.

BACKGROUND: Previous studies have indicated that single nucleotide polymorphisms (SNPs) of the interleukin-17 (IL-17) gene are associated with an increased risk of gastric cancer. However, the findings were inconsistent. MATERIALS AND METHODS: To provide a more reliable estimation of the association between SNPs in the IL-17 gene and the susceptibility to gastric cancer, we searched PubMed, CNKI, and Wan Fang databases and selected finally six studies covering 2,366 cases and 3,205 controls to perform a meta-analysis. RESULTS: Statistical analyses showed that an rs2275913 polymorphism within the IL-17A gene was significantly associated with an increased risk of gastric cancer using a generalized odds ratio (ORG, a model-free approach). Moreover, we also found that the 'A' allele carriers of IL-17A rs2275913 had a significant link with clinicopathological features. However, no significant positive signals were observed in the association analysis of the rs3748067 and rs763780 polymorphisms with the risk of gastric cancer in IL-17A and IL-17F, respectively. CONCLUSIONS: Despite some limitations, the present meta-analysis provided a more precise estimation of the relationship between the IL-17 gene SNPs and gastric cancer risk compared with individual studies.

A small-molecule FRET reporter for the real-time visualization of cell-surface proteolytic enzyme functions.

Real-time imaging of cell-surface-associated proteolytic enzymes is critical to better understand their performances in both physiological and pathological processes. However, most current approaches are limited by their complexity and poor membrane-anchoring properties. Herein, we have designed and synthesized a unique small-molecule fluorescent probe, which combines the principles of passive exogenous membrane insertion and Förster resonance energy transfer (FRET) to image cell-surface-localized furin-like convertase activities. The membrane-associated furin-like enzymatic cleavage of the peptide probe leads to an increased fluorescence intensity which was mainly localized on the plasma membrane of the furin-expressed cells. This small-molecule fluorescent probe may serve as a unique and reliable reporter for real-time visualization of endogenous cell-surfaceassociated proteolytic furin-like enzyme functions in live cells and tissues using one-photon and two-photon microscopy.

Conjugated-polymer-based red-emitting nanoparticles for two-photon excitation cell imaging with high contrast.

Two-photon fluorescence microscopy is a widely used noninvasive bioimaging technique because of unique advantages such as a large penetration depth and 3D mapping capability. Ideal two-photon fluorophores require large two-photon absorption cross sections and red emission with high quantum yields. Here we report red-emitting-dye-doped conjugated polymer nanoparticles that display high two-photon excitation brightness. In these nanoparticles, conjugated polymer (PFV) was chosen as a two-photon light-harvesting material, and red-emitting dyes (MgPc and Nile red) were chosen as the energy acceptors and red-emitting materials. Two-photon excitation fluorescence of MgPc and Nile red was enhanced by up to ∼53 and ∼240 times, respectively. We have successfully demonstrated the application of these conjugated polymer-based nanoparticles in two-photon excitation cancer cell imaging with an excellent contrast ratio. This concept could become a general approach to the preparation of two-photon excitation red-emitting materials for deep-tissue live-cell imaging with high contrast.

Gold nanorod enhanced two-photon excitation fluorescence of photosensitizers for two-photon imaging and photodynamic therapy.

Plasmon enhancement of optical properties is both fundamentally important and appealing for many biological and photonic applications. Although metal-enhanced two-photon excitation fluorescence has been demonstrated in the solid substrates, there is no report on metal enhanced overall two-photon excitation fluorescence in the colloid system. Here we systematically investigated gold nanorod enhanced one- and two-photon excitation fluorescence of a porphyrin molecule, T790. The separation distance between the metal core and T790 was varied by adjusting the silica shell thickness from 13 to 42 nm. One- and two-photon excitation fluorescence intensities of T790 were found to strongly depend on the thickness of silica shell that separates gold nanorod and T790. The optimum one- and two-photon excitation fluorescence enhancement was found to occur at shell thicknesses of 34 and 20 nm, with enhancement factors of 2.1 and 11.8, respectively. Fluorescence lifetime of T790 steadily decreased as the shell thickness decreased. The observed two-photon excitation fluorescence enhancement is ascribed to a combination effect of local electric field amplification and competition between increased radiative and non-radiative decay rates. Core-shell nanoparticles that displayed enhanced two-photon excitation fluorescence were also found to exhibit significantly improved singlet oxygen generation capability under two-photon excitation. The applications of these nanoparticles as effective agents for two-photon cell imaging and nano-photosensitizers for two-photon photodynamic therapy with improved efficiency have also been demonstrated in HepG2 cancer cells. The combined advantages of enhanced two-photon excitation fluorescence and two-photon induced singlet oxygen generation make these core-shell nanoparticles as attractive agents for two-photon imaging guided two-photon photodynamic therapy.