Cxxc finger protein 1 maintains homeostasis and function of intestinal group 3 innate lymphoid cells with aging.

Aging is accompanied by homeostatic and functional dysregulation of multiple immune cell subsets. Group 3 innate lymphoid cells (ILC3s) constitute a heterogeneous cell population that plays pivotal roles in intestinal immunity. In this study, we found that ILC3s in aged mice exhibited dysregulated homeostasis and function, leading to bacterial and fungal infection susceptibility. Moreover, our data revealed that the enrichment of the H3K4me3 modification in effector genes of aged gut CCR6+ ILC3s was specifically decreased compared to young mice counterparts. Disruption of Cxxc finger protein 1 (Cxxc1) activity, a key subunit of H3K4 methyltransferase, in ILC3s led to similar aging-related phenotypes. An integrated analysis revealed Kruppel-like factor 4 (Klf4) as a potential Cxxc1 target. Klf4 overexpression partially restored the differentiation and functional defects seen in both aged and Cxxc1-deficient intestinal CCR6+ ILC3s. Therefore, these data suggest that targeting intestinal ILC3s may provide strategies to protect against age-related infections.

Advances in liquid biopsy-based markers in NSCLC.

Lung cancer is the second most-frequently occurring cancer and the leading cause of cancer-associated deaths worldwide. Non-small cell lung cancer (NSCLC), the most common type of lung cancer is often diagnosed in middle or advanced stages and have poor prognosis. Diagnosis of disease at an early stage is a key factor for improving prognosis and reducing mortality, whereas, the currently used diagnostic tools are not sufficiently sensitive for early-stage NSCLC. The emergence of liquid biopsy has ushered in a new era of diagnosis and management of cancers, including NSCLC, since analysis of circulating tumor-derived components, such as cell-free DNA (cfDNA), circulating tumor cells (CTCs), cell-free RNAs (cfRNAs), exosomes, tumor-educated platelets (TEPs), proteins, and metabolites in blood or other biofluids can enable early cancer detection, treatment selection, therapy monitoring and prognosis assessment. There have been great advances in liquid biopsy of NSCLC in the past few years. Hence, this chapter introduces the latest advances on the clinical application of cfDNA, CTCs, cfRNAs and exosomes, with a particular focus on their application as early markers in the diagnosis, treatment and prognosis of NSCLC.

The Transcription Factor ThPOK Regulates ILC3 Lineage Homeostasis and Function During Intestinal Infection.

Innate lymphoid cells (ILCs) have been identified as a heterogeneous population of lymphocytes that mirrors the cytokine and transcriptional profile of adaptive T cells. The dynamic balance between key transcription factors determines the heterogeneity, plasticity, and functions of ILC subsets. The transcription factor ThPOK is highly conserved in biological evolution and exerts pivotal functions in the differentiation of T cells. However, the function of ThPOK in ILC3s has not been identified. Here, we found that ThPOK regulated the homeostasis of ILC3s, as mice lacking ThPOK showed decreased NKp46+ ILC3s and increased CCR6- NKp46- ILC3s. ThPOK-deficient mice were more sensitive to S. typhimurium infection due to the impaired IFN-γ secretion of NKp46+ ILC3s. Furthermore, ThPOK participates in ILC3-mediated control of C. rodentium infection by negatively regulating IL-17A secretion. ThPOK preserves the identity of NKp46+ ILC3s by repressing RORγt, which indirectly releases T-bet expression. On the molecular level, ThPOK directly binds to Rorc and Il23r to restrain their expression which further modulates IL-17A secretion. Collectively, our analysis revealed a critical role of ThPOK in the homeostasis and functions of ILC3 subsets.

Label-free detection of biomarker alpha fetoprotein in serum by ssDNA aptamer functionalized magnetic nanoparticles.

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors in the clinic, with the characteristics of occult onset, rapid progression, and high degree of malignancy. Alpha fetoprotein (AFP) is the most important biomarker of HCC, which is widely used in early screening, diagnosis, and prognosis observation. A series of immunoassays have been developed and frequently used in the detection of AFP based on antibodies. Unfortunately, the shortcomings of antibodies, such as thermal unstable and fluctuant activity by batches, lead to the inaccuracy in the detection of AFP. In this study, aptamers instead of antibodies were adopted as the specific recognition element for AFP, aiming to seek an alternative strategy to immunoassays. An AFP-specific ssDNA aptamer was grafted to magnetic nanoparticles (Fe3O4@SiO2) via avidin-biotin interaction, and the resultant aptamer functionalized magnetic nanoparticles (Ap-MNPs) were adequately characterized and tested. The Ap-MNPs in solution exhibited a fast response to the outer magnetic field, and can be completely separated in several minutes. It was found that Ap-MNPs have good specificity to the target AFP, as the recovery of AFP (87.0%) was much higher than the competitive proteins IgG (38.9%), HSA (18.5%), and FIB (11.4%). A convenient and efficient label-free detection method of AFP in serum was developed based on Ap-MNPs in combination with high-performance liquid chromatography. The linearity of this method was over a range of 1-50 µg ml-1 with a correlation coefficient of 0.9999, and the limit of detection was 0.27 µg ml-1. This study indicated that aptamers are an ideal tool for the recognition and detection of biomarkers, and thus will find wide applications in clinical practice.

The fluorescence and resonance Rayleigh scattering spectral study and analytical application of cerium (IV) and cefoperazone system.

In weak acidic medium of pH3.5-5.6, Ce(IV) can be reduced by cefoperazone (CPZ) to be Ce(III), which further combined with CPZ to form complex Ce(OH)3CPZ. This complex not only has higher fluorescence than Ce(III), but also results in significant increase of resonance Rayleigh scattering (RRS), second order scattering (SOS) and frequency doubling scattering (FDS). The wavelengths of maximum fluorescence exciting and emission are located at 356 nm/349 nm, while the maximum wavelengths of RRS, SOS and FDS are at 312 nm, 550 nm and 390 nm, respectively. The intensity of fluorescence and scattering are all linear with the concentration of CPZ in certain conditions. The detection limit of most sensitive RRS method for CPZ is 2.1 ng mL(-1). The optimum conditions for detecting CPZ using RRS method are investigated. The effect of co-existing substances shows that the method has excellent selectivity, especially since other cephalosporins don't have similar reactions. Therefore, it can be achieved to determine CPZ in cephalosporins selectively. The paper also focuses on the reaction mechanism, the consistent and contracture of the resultant. The reasons for enhanced intensity are presumed in the meantime.

Resonance Rayleigh scattering spectra of Cu2+-adenine-WO4(2-) system and its analytical application.

In pH 6.6-7.2 Tris-HCl buffer, Cu(2+) could react with adenine (A) to form a 1:1 coordination cation [CuA](2+), which only resulted in minor change of the absorption spectrum. However, when this cation further combined with WO(4)(2-) to form a 1:1 ternary ion-association complex [CuA]WO(4), the absorption spectrum changed a lot, and the resonance Rayleigh scattering (RRS), second-order scattering (SOS) and frequency doubling scattering (FDS) enhanced significantly. The maximum wavelengths of RRS, SOS and FDS were located at 310, 592 and 395 nm, respectively. The enhanced intensities of the three methods were proportional to the concentration of adenine in certain ranges, and the detection limit of the most sensitive RRS method was 7.4 × 10(-9) mol L(-1) (1.0 ng mL(-1)), indicating that this method could detect trace adenine. In this work, the optimum reaction conditions and the influencing factors have been studied, some potential interferences and the composition of the ion-association complex have been investigated. Meanwhile, the construction of the product and the reaction mechanism have been investigated by atomic force microscopy, transmission electron microscope and quantum chemical calculation. Accordingly, a novel RRS method for determination of adenine has been proposed and applied to detect adenine in real samples with satisfactory results.

Interaction of adenine with palladium chloride, determination of adenine via resonance Rayleigh scattering method.

In pH 1.7-3.5 acid medium, palladium chloride could react with adenine (A) to form a ternary complex of [PdCl(2)·A], which would self-aggregate to form uniformly dispersed nanoparticles-[PdCl(2)·A](n) with an average size of 42 nm through the squeezing effect of aqueous phase and van der Waals force. This resulted in an enhancement of resonance Rayleigh scattering (RRS), second-order scattering (SOS) and frequency doubling scattering (FDS). The maximum wavelengths were located at 311 nm, 611 nm and 395 nm, respectively. The scattering intensities of the three methods were proportional to the concentration of adenine in certain ranges, and the detection limit of the most sensitive RRS method was 5.4×10(-9) mol L(-1) (0.73 ng mL(-1)). The experimental conditions were optimized and effects of coexisting substances were evaluated. The method showed excellent selectivity because a certain amount of other nucleobase, nucleoside or nucleotide would not influence the measurement. Accordingly, a novel rapid, convenient, sensitive and selective RRS method for determination of adenine was proposed and applied to detect adenine in tablet and hydrolyzates of ctDNA samples with satisfactory results. The shape of nanoparticles was characterized by atomic force microscopy. The reaction mechanism and the reasons for enhancement of scattering were discussed by infrared spectra, quantum chemical calculations and absorption spectroscopy.