Integrated single-cell analysis defines the epigenetic basis of castration-resistant prostate luminal cells.

Understanding prostate response to castration and androgen receptor signaling inhibitors (ARSI) is critical to improving long-term prostate cancer (PCa) patient survival. Here, we use a multi-omics approach on 229,794 single cells to create a mouse single-cell reference atlas for interpreting mouse prostate biology and castration response. Our reference atlas refines single-cell annotations and provides a chromatin context, which, when coupled with mouse lineage tracing, demonstrates that castration-resistant luminal cells are distinct from the pre-existent urethra-proximal stem/progenitor cells. Molecular pathway analysis and therapeutic studies further implicate AP1 (JUN/FOS), WNT/ß-catenin, FOXQ1, NF-κB, and JAK/STAT pathways as major drivers of castration-resistant luminal populations with relevance to human PCa. Our datasets, which can be explored through an interactive portal (https://visportal.roswellpark.org/data/tang/), can aid in developing combination treatments with ARSI for advanced PCa patients.

Hydrazone-linked covalent organic framework functionalized with cysteine as a fluorescence sensor and Exploration of paper chip for p-nitrophenol detection.

The large use and emission of p-nitrophenol (p-NP) seriously pollute the environment and endanger human health. In this work, a hydrazone-linked fluorescent covalent organic framework (BATHz-COF) was simply synthesized at room temperature and covalently linked N-acetyl-L-cysteine (NALC) via the "thiol-ene" click reaction, where carboxyl groups were introduced to improve dispersion and fluorescence intensity. As a rapid, good selectivity and reusability fluorescence sensor, the obtained COF-NALC has been used for quantitative analysis of p-NP predicated on the internal filtering effect (IFE). Under optimal conditions, COF-NALC enabled quantitative detection of p-NP with a linear range of 5-50 µM and the detection limit was 1.46 µM. The application of COF-NALC to the detection of p-NP in river water samples was successful, and the satisfactory recoveries were 98.0%-109.3%. Furthermore, the fluorescent COF paper chips constructed by in situ growth were combined with a smartphone to build a visual platform for the quick and real-time detection of p-NP, providing an excellent illustration for the development of intelligent fluorescence sensing in environmental analysis.

Supramolecular Interaction Modulation in Thermosensitive Composites: Enantiomeric Recognition and Chiral Site Regeneration.

Herein, a novel strategy was implemented to modulate the supramolecular interaction between enantiomers and chiral recognition sites (CRSs), effectively resolving the issue of CRS saturation. Randomly methylated-ß-cyclodextrin (Rm-ß-CD) was used as the CRS (host molecule), and polymerized ionic liquids [poly([vbim]TFSI)] were used as the supramolecular modulator (guest molecule), which self-assembled to generate thermosensitive supramolecular host/guest complexes. The enantiomeric binding capacity and enantioselectivity of chiral separation systems centered on supramolecular host-guest complexes are characterized by a high degree of temperature dependence. Poly([vbim]TFSI) bonded to Rm-ß-CD at temperatures between 17 °C ± 3 and 50 °C ± 3 °C, and the binding free energy difference (|ΔΔG|) between the (S)- and (R)-enantiomer was 0.55. Conversely, poly([vbim]TFSI detached from Rm-ß-CD at temperatures >50 °C ± 3 °C or <17 °C ± 3 °C, and |ΔΔG| between (S)- and (R)-enantiomer was 0.03. The |ΔΔG| value of the (R)-enantiomer can reach 0.86 in two temperature intervals. Therefore, the binding of poly([vbim]TFSI) to Rm-ß-CD afforded the favorable separation of four racemic sample mixtures: mandelic acid (e.e.% = 61.3%), ibuprofen (e.e.% = 21.6%), warfarin (e.e.% = 14.9%), and naproxen (e.e% = 18.2%). The detachment of poly([vbim]TFSI) from Rm-ß-CD released the enantiomer bound to CRSs. The decomplexation of mandelic acid reached 75.1%.

Construction and Application of a Pepsin-Functionalized Covalent Organic Framework with Prominent Chiral Recognition Ability.

The stable Pepsin@covalent organic framework (Pepsin@COF) were constructed base on matching COF pore diameter to pepsin dimension. It exhibits excellent chiral recognition capabilities (e. e. % up to 62.63 %) and potential for enantioseparation. Furthermore, a positive correlation between the immobilized enzyme activity and chiral recognition was revealed, offering insights for the design of biocatalytic nanosystems in chiral separation.

Scalable expansion of human pluripotent stem cells under suspension culture condition with human platelet lysate supplementation.

The large-scale production of human pluripotent stem cells (hPSCs), including both embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs), shows potential for advancing the translational realization of hPSC technology. Among multiple cell culture methods, suspension culture, also known as three-dimensional (3D) culture, stands out as a promising method to fulfill the large-scale production requirements. Under this 3D culture condition, cell expansion and the preservation of pluripotency and identity during long-term culture heavily relies on the culture medium. However, the xenogeneic supplements in culture medium remains an obstacle for the translation of cell and gene therapy applications from bench to bedside. Here, we tested human platelet lysate (hPL), a xeno-free and serum-free biological material, as a supplement in the 3D culture of hPSCs. We observed reduced intercellular variability and enhanced proliferation in both hESC and hiPSC lines. These cells, after extended culture in the hPL-supplemented system, maintained pluripotency marker expression, the capacity to differentiate into cells of all three germ layers, and normal karyotype, confirming the practicability and safety of hPL supplementation. Furthermore, through RNA-sequencing analysis, we found an upregulation of genes associated with cell cycle regulations in hPL-treated cells, consistent with the improved cellular division efficiency. Taken together, our findings underscore the potential of hPL as a xeno-free and serum-free supplement for the large-scale production of hPSCs, which holds promise for advancing clinical applications of these cells.

Carboxy-Functionalized Covalent Organic Framework as a Carrier for Lipase Immobilization and Its Application in Inhibitors Screening.

Covalent organic frameworks (COFs) with large specific surface areas, high porosity, good stability, and designable structure are promising carriers for immobilized enzymes. It is important to explore lipase inhibitors from natural foods as lipase inhibitors are closely related to the treatment of obesity. In this work, a carboxyl functionalized covalent organic framework (TpBD-3COOH) was prepared by solvothermal method for covalent immobilization of porcine pancreatic lipase (PPL) and obtained the enzyme-decorated COF (PPL@COF). The immobilized lipase showed wider pH and temperature tolerance with the same optimal pH and temperature of 7.5 and 50 ℃ compared to free lipase. After 6 successive reuses, the PPL@COF maintained 53.0% of its original activity. Immobilized lipase also displayed enhanced storage stability (55.4% after 14 days at 4 ℃). When p-nitrophenyl acetate was applied as the substrate, the calculated Michaelis constant was 3.57 mM and the half maximal inhibitory concentration of orlistat was 3.20 µM. Finally, the PPL@COF was used for enzyme inhibitors screening from natural foods combined with UV spectrophotometry, and Hawthorn was screened for excellent lipase inhibitory activity.

Resuscitation of out-of-hospital cardiac arrest in China: A systematic review and Utstein-style data analysis based on the Chain of Survival.

AIM: Out-of-hospital cardiac arrest (OHCA) contributes to substantial mortality, but its resuscitation status in China is unknown. We aimed to describe and analyze out-of-hospital cardiac arrest in terms of Chain of Survival. METHODS: We systematically collected Utstein-style publications. Scenarios were prespecified, including either emergency medical service (EMS) assessing and attending cardiac arrest, resuscitation attempted by a bystander, resuscitation attempted by EMS, or in-hospital treatment. Random-effect models were used in a meta-analysis to pool rate ratios (RRs) with 95% confidence intervals (CIs) from multiple cohorts. RESULTS: We analyzed 59 Chains involving 233,376 Chinese patients. The median rate of survival to discharge (interquartile range) was 0.35 % (0.06 %-0.61 %), 3.66 % (3.06 %-3.85 %), 1.23 % (0.57%-1.36%), and 2.73% (2.04%-3.42%) for four scenarios. The rate was significantly higher for bystander resuscitation than for EMS (P = 0.025) or in-hospital treatment (P = 0.301). However, only 4.8 % (1.6 %-8.2 %) of patients received bystander resuscitation, with no bystander defibrillation and a median response time of 9-15 minutes for EMS. Compared with controls without witnesses, arrest being witnessed and with bystander resuscitation increased rates of survival to discharge by 1.97 (I2 = 0, P for I2 = 0.583; pooled RR 2.97; 95% CI 1.47-6.02) and 6.79 (I2 = 0, P for I2 = 0.593; pooled RR 7.79; 95 % CI 3.40-17.84) times, following a markedly increasing trend. CONCLUSIONS: A low probability of first aid at multiple points is linked to poor survival following OHCA. It is essential to strengthen front links in the Chain of Survival in China, including among witnesses, bystanders, and emergency response.

Self-Calibrating Lanthanide Infinite Coordination Polymer Constructs Fluorescent Probes: A Sensitive Approach for Early Diagnosis of Hepatocellular Carcinoma and Environmental Analysis.

The development of a self-calibrating ratio fluorescence probe without the need for additional substrates is a major advancement in biosensing. In this study, at room temperature, a self-calibrating infinite coordination polymer (SSA-Tb-ATP ICPs) has been proposed by self-assembling adenosine triphosphate (ATP) with 5-sulfosalicylic acid (SSA) and Tb3+. Due to the antenna effect, SSA-Tb-ATP ICPs exhibited strong green fluorescence emission of Tb3+ (at 547 nm) and blue fluorescence emission of SSA (at 407 nm). This material offers several advantages over existing detection methods, including simplicity of synthesis and exceptional sensitivity. Our self-calibrating SSA-Tb-ATP ICPs demonstrated excellent performance in detecting alkaline phosphatase (ALP) and phosphate (Pi) in both serum and environmental samples with detection limits of 0.076 U/L and 0.025 µM, respectively. Moreover, we successfully employed the SSA-Tb-ATP ICPs to perform cellular imaging of ALP in both hepatocellular carcinoma cells (HepG2) and normal liver cells (LO2), representing a significant advancement in ALP detection and imaging. The simplicity of the synthesis and high sensitivity make this probe a promising tool for early diagnosis of hepatocellular carcinoma in clinical settings and environment analysis.

Construction of chiral capillary electrochromatography microsystems based on Aspergillus sp. CM96.

Novel chiral capillary electrochromatography (CEC) microsystems were constructed based on Aspergillus sp. CM96. As a newly discovered intrinsic characteristic of the cell, cell chirality occupies an essential position in life evolution. Aspergillus sp. CM96 spore (CM96s) was chosen as a proof of concept to develop chiral capillary columns. Interestingly, various types of amino acid (AA) enantiomers were baseline separated under the optimized conditions. Furthermore, the time-dependent chiral interactions between AAs and CM96s were explored in a wider space. Pectinases generated from Aspergillus sp. CM96 fermentation were immobilized onto graphene oxide-functionalized capillary silica monoliths for separating AA enantiomers. Molecular docking simulations were performed to explore chiral separation mechanisms of pectinase for AA enantiomers. These results indicated that Aspergillus sp. CM96-based CEC microsystems have a significant advantage for chiral separation.

A promising tool for clinical diagnostics: Dual-emissive carbonized polymer dots based cross-linking enhanced emission for sensitive detection of alkaline phosphatase and butyrylcholinesterase.

Compared with single signal readout, dual-signal readout commendably corrects the impact of systematic or background error, achieving more accurate results for the diagnosis of many diseases. This work aimed to design and prepare dual-emissive fluorescent probes for the construction of ratiometric fluorescence biosensors to detect liver disease biomarkers. Sodium alginate (SA) with numerous potential sub-fluorophores and active sites and 4,4',4'',4'''-(porphine-5,10,15,20-tetrayl) tetrakis (benzoic acid) (TCPP) with macrocyclic conjugated structures were introduced to prepare the carbonized polymer dots (CPDs) with red/blue dual emission based on the cross-linking enhanced emission (CEE) effect and the luminescence of macrocyclic conjugated structures. The ratiometric fluorescence sensing systems were constructed by integrating the specific response of CPDs to Cu2+ and the affinity difference of Cu2+ to substrates or products of enzymes. The sensing systems, CPDs/Cu2+/PPi and CPDs/Cu2+/BTCh, were designed to detect liver disease biomarkers, alkaline phosphatase (ALP) and butyrylcholinesterase (BChE), respectively. The limit of detection for ALP and BChE was 0.35 U/L and 0.19 U/L, respectively. The proposed sensors were successfully applied to human serum samples from different health stages with satisfactory recoveries. These results demonstrate the successful design of a novel dual-emissive fluorescent probe and provide a feasible strategy for clinical detection.

Chirality-Controlled Mercapto-ß-cyclodextrin Covalent Organic Frameworks for Selective Adsorption and Chromatographic Enantioseparation.

Chiral covalent organic frameworks (CCOFs) benefit from superior stability, abundant chiral environment, and homogeneous pore configuration. In its constructive tactics, only the post-modification method allows for the integration of supramolecular chiral selectors into achiral COFs. Here, the finding utilizes 6-deoxy-6-mercapto-ß-cyclodextrin (SH-ß-CD) as chiral subunits and 2,5-dihydroxy-1,4-benzenedicarboxaldehyde (DVA) as the platform molecule to synthesize chiral functional monomers through thiol-ene click reactions and directly establish ternary "pendant-type" SH-ß-CD COFs. The chiral site density on SH-ß-CD COFs was regulated by changing the proportion of chiral monomers to obtain an optimal construction strategy and remarkably improve the ability of chiral separation. SH-ß-CD COFs were coated on the inner wall of the capillary in a covalently bound manner. The prepared open tubular capillary was achieved for the separation of six chiral drugs. By combining the outcomes of selective adsorption and chromatographic separation, we observed the higher density of chiral sites in the CCOFs, and poorer results were achieved. From the perspective of spatial conformational distribution, we interpret the variation in the performance of these chirality-controlled CCOFs for selective adsorption and chiral separation.

Integrated single-cell analysis defines the epigenetic basis of castration-resistant prostate luminal cells.

Understanding prostate response to castration and androgen receptor signaling inhibitors (ARSI) is critical to improving long-term prostate cancer (PCa) patient survival. Here we use a multi-omics approach on 229,794 single cells to create a mouse single-cell reference atlas better suited to interpreting mouse prostate biology and castration response. Our reference atlas refines single-cell annotations and provides chromatin context, which, when coupled with mouse lineage tracing demonstrates that the castration-resistant luminal cells are distinct from the pre-existent urethra-proximal stem/progenitor cells. Molecular pathway analysis and therapeutic studies further implicate JUN/FOS, WNT/B-Catenin, FOXQ1, NFkB, and JAK/STAT pathways as the major drivers of castration-resistant luminal populations with high relevance to human PCa. Importantly, we demonstrate the utility of our datasets, which can be explored through an interactive portal (https://visportal.roswellpark.org/data/tang/), to aid in developing novel combination treatments with ARSI for advanced PCa patients.

Dual-mode strategy for 2,6-dipicolinic acid detection based on the fluorescence property and peroxidase-like activity inhibition of Fe-MIL-88NH2.

This work designed a fluorometric/colorimetric dual-mode sensor for detecting 2,6-dipicolinic acid (DPA) based on the blue emission property and peroxidase-like activity of Fe-MIL-88NH2. The fluorescence of Fe-MIL-88NH2 was obviously turned off by Cu2+, but DPA was able to bring it back because it has a strong chelate bond with Cu2+. Fe-MIL-88NH2 also displayed high peroxidase-like activity, which accelerated the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) to the blue oxidation product (oxTMB) when H2O2 was present. When DPA was added, it efficiently inhibited the peroxidase-like activity of Fe-MIL-88NH2, causing less oxTMB and less absorbance at 652 nm. The fluorescence recovery of Fe-MIL-88NH2 and the change in absorbance at 652 nm were used as analytical signals for dual-mode detection of DPA. The linear responses in the range of 10-60 µM and 60-160 µM were achieved for the fluorometric mode, and the limit of detection (LOD) was 1.46 µM. The respective values of linear range and LOD for the colorimetric mode were 5-25 µM and 3.00 µM, respectively. In summary, the dual-mode testing strategy successfully detected DPA in aqueous environmental samples, suggesting great potential in disease prevention and environmental analysis.

Immobilized glucose oxidase on hierarchically porous COFs and integrated nanozymes: a cascade reaction strategy for ratiometric fluorescence sensors.

Covalent organic frameworks (COFs) with uniform porosity, good stability, and desired biocompatibility can function as carriers of immobilized enzymes. However, the obstructed pores or partially obstructed pores have hindered their applicability after loading enzymes. In this study, the hierarchical COFs were prepared as an ideal support to immobilize glucose oxidase (GOD) and obtain GOD@COF. The hierarchical porosity and porous structures of COFs provided sufficient sites to immobilize GOD and increased the rate of diffusion of substrate and product. Moreover, N,Fe-doped carbon dots (N,Fe-CDs) with peroxidase-like activity were introduced to combine with GOD@COF to construct an enzyme-mediated cascade reaction, which is the basis of the sensor GOD@COF/N,Fe-CDs. The sensor has been successfully built and applied to detect glucose. The limit of detection was 0.59 µM for determining glucose with the proposed fluorescence sensor. The practicability was illustrated by detecting glucose in human serum and saliva samples with satisfactory recoveries. The proposed sensor provided a novel strategy that introduced COF-immobilized enzymes for cascade reactions in biosensing and clinical diagnosis.

Bifunctional Fe@PCN-222 nanozyme-based cascade reaction system: Application in ratiometric fluorescence and colorimetric dual-mode sensing of glucose.

This work innovatively integrated the peroxidase-mimicking activity and red emission property of Fe@PCN-222 framework, designed a cascade reaction system for dual-mode glucose sensing. The Fe3+ doping significantly improved the catalytic activity of Fe@PCN-222 that can oxidize the substrate o-phenylenediamine (OPD) to generate diminophenazine (DAP) with emission at 566 nm in the presence of H2O2. Similarly, the Fe@PCN-222 was used to catalyze the colorless TMB to produce blue oxidized TMB (oxTMB) showed absorption at 652 nm. When coupled with glucose oxidase (GOx), the linear ranges of ratiometric fluorescence mode and colorimetric mode for glucose sensing were 1-100 and 10-300 µM, respectively. And the limits of detection (LOD) of 0.78 and 2.41 µM for two modes were obtained, respectively. In addition, the practicability of Fe@PCN-222 nanozyme-based cascade reaction system for detection of glucose in human serum and saliva samples was successfully investigated. It is of great importance to integrate more functions into one skeleton to achieve dual-mode and optimal-performance sensing for expanding potential applications.

Enhanced Chiral Recognition Abilities of Cyclodextrin Covalent Organic Frameworks via Chiral/Achiral Functional Modification.

ß-Cyclodextrin covalent organic frameworks (ß-CD COFs) show great potential in enantioseparation due to their uniformly distributed chiral recognition sites and good chemical stability. The hydroxyl and amino groups of ß-CD COFs enable facile post-modification to introduce the desired functionality into the frameworks. In this study, we perform post-modification of ß-CD COFBPDA with 1,4-butane sultone and [(3R,4R)-4-acetyloxy-2,5-dioxooxolan-3-yl] acetate to construct two kinds of novel functional ß-CD COFs. The capillary columns prepared with these two functional ß-CD COFs separated chiral dihydropyridines and fluoroquinolones with excellent selectivity and repeatability in capillary electrochromatography, while ß-CD COFBPDA-modified capillary columns did not present the chiral recognition ability for these drugs. The mechanism of chiral recognition and the enhanced enantioselectivity of functional ß-CD COFs were further demonstrated by molecular docking simulation. The divergent chiral separation performances of ß-CD COFs suggest that the introduction of functional groups enables the modification of ß-CD COF properties and tuning of its chiral recognition abilities for the diversity of enantioseparation.

LRIG1, a regulator of stem cell quiescence and a pleiotropic feedback tumor suppressor.

LRIG1, leucine-rich repeats and immunoglobulin-like domains protein 1, was discovered more than 20 years ago and has been shown to be downregulated or lost, and to function as a tumor suppressor in several cancers. Another well-reported biological function of LRIG1 is to regulate and help enforce the quiescence of adult stem cells (SCs). In both contexts, LRIG1 regulates SC quiescence and represses tumor growth via, primarily, antagonizing the expression and activities of ERBB and other receptor tyrosine kinases (RTKs). We have recently reported that in treatment-naïve human prostate cancer (PCa), LRIG1 is primarily regulated by androgen receptor (AR) and is prominently overexpressed. In castration-resistant PCa (CRPC), both LRIG1 and AR expression becomes heterogeneous and, frequently, discordant. Importantly, in both androgen-dependent PCa and CRPC models, LRIG1 exhibits tumor-suppressive functions. Moreover, LRIG1 induction inhibits the growth of pre-established AR+ and AR- PCa. Here, upon a brief introduction of the LRIG1 and the LRIG family, we provide an updated overview on LRIG1 functions in regulating SC quiescence and repressing tumor development. We further highlight the expression, regulation and functions of LRIG1 in treatment-naïve PCa and CRPC. We conclude by offering the perspectives of identifying novel cancer-specific LRIG1-interacting signaling partners and developing LRIG1-based anti-cancer therapeutics and diagnostic/prognostic biomarkers.

Facile separation of enantiomers via covalent organic framework bonded stationary phase.

Covalent organic frameworks (COFs), a type of crystalline polymers, have attracted increasing interest because of their controllability of geometry and functionality. Featuring infinitely extended networks and tremendous interaction sites, COFs emerge as a potential platform for separation science. Here, a novel chiral COF (ß-CD COFBPDA) constructed by the imine condensation of 4,4'-biphenyldicarboxaldehyde and heptakis(6-amino-6-deoxy)-ß-cyclodextrin was introduced into an electrochromatographic system via a photopolymerization method and applied to the separation of enantiomers. The structure and properties of as-synthesized ß-CD COFBPDA were investigated by powder X-ray diffraction (PXRD) patterns, Fourier transform infrared (FT-IR) spectroscopy, thermogravimetric analysis (TGA), and N2adsorption-desorption isotherms. It was proved that ß-CD COFBPDA was provided with larger pore size and BET surface area. The ß-CD COFBPDA coating endowed the chiral stationary phase with superior three-dimensional orientation, and realized satisfactory separation with improved selectivity and column efficiency for a dozen racemic drugs. Under the optimized conditions, homatropine, ondansetron, metoprolol, terbutaline, tulobuterol, and promethazine were all baseline separated with resolution values of 2.24, 2.03, 1.65, 1.62, 1.60, and 1.58, respectively. The results indicate the high perspective of COF modified stationary in enantioseparation.

Chiral Carboxyl-Functionalized Covalent Organic Framework for Enantioselective Adsorption of Amino Acids.

Combining chiral molecules and covalent organic frameworks (COFs) with ordered pores can result in chiral COFs, which may be employed in the fields of chiral chemistry. In this study, we constructed a novel carboxyl-functionalized COF TpBD-3COOH first and then integrated a chiral molecule, heptakis (6-amino-6-deoxy)-ß-CD (Am7CD), into TpBD-3COOH to obtain TpBD-Am7CD. Comparing TpBD-3COOH with TpBD-Am7CD, it could be observed that chiral selectivities improved significantly after decorating TpBD-3COOH with Am7CD in the adsorption experiment. Meanwhile, TpBD-Am7CD exhibited similar chiral selective abilities as the previously reported ß-CD COF when it was employed to adsorb amino acid enantiomers, while the content of Am7CD was much less than that in ß-CD COF. This indicated that the framework of TpBD-3COOH enhanced the chiral selective ability of Am7CD, which would be a great chiral carrier material for further application. This work provides a new insight for establishing materials for application in the chiral recognition field. Besides, TpBD-Am7CD resulted in an insoluble powder, which endowed this material with the possibility to be an adsorbent and a stationary phase in chromatography.