Applications of Intravital Imaging in Cancer Immunotherapy.

Currently, immunotherapy is one of the most effective treatment strategies for cancer. However, the efficacy of any specific anti-tumor immunotherapy can vary based on the dynamic characteristics of immune cells, such as their rate of migration and cell-to-cell interactions. Therefore, understanding the dynamics among cells involved in the immune response can inform the optimization and improvement of existing immunotherapy strategies. In vivo imaging technologies use optical microscopy techniques to visualize the movement and behavior of cells in vivo, including cells involved in the immune response, thereby showing great potential for application in the field of cancer immunotherapy. In this review, we briefly introduce the technical aspects required for in vivo imaging, such as fluorescent protein labeling, the construction of transgenic mice, and various window chamber models. Then, we discuss the elucidation of new phenomena and mechanisms relating to tumor immunotherapy that has been made possible by the application of in vivo imaging technology. Specifically, in vivo imaging has supported the characterization of the movement of T cells during immune checkpoint inhibitor therapy and the kinetic analysis of dendritic cell migration in tumor vaccine therapy. Finally, we provide a perspective on the challenges and future research directions for the use of in vivo imaging technology in cancer immunotherapy.

Ultrafast bisulfite sequencing detection of 5-methylcytosine in DNA and RNA.

Bisulfite sequencing (BS-seq) to detect 5-methylcytosine (5mC) is limited by lengthy reaction times, severe DNA damage, overestimation of 5mC level and incomplete C-to-U conversion of certain DNA sequences. We present ultrafast BS-seq (UBS-seq), which uses highly concentrated bisulfite reagents and high reaction temperatures to accelerate the bisulfite reaction by ~13-fold, resulting in reduced DNA damage and lower background noise. UBS-seq allows library construction from small amounts of purified genomic DNA, such as from cell-free DNA or directly from 1 to 100 mouse embryonic stem cells, with less overestimation of 5mC level and higher genome coverage than conventional BS-seq. Additionally, UBS-seq quantitatively maps RNA 5-methylcytosine (m5C) from low inputs of mRNA and allows the detection of m5C stoichiometry in highly structured RNA sequences. Our UBS-seq results identify NSUN2 as the major 'writer' protein responsible for the deposition of ~90% of m5C sites in HeLa mRNA and reveal enriched m5C sites in 5'-regions of mammalian mRNA, which may have functional roles in mRNA translation regulation.

Predicting the Sauter Mean Diameter of Swirl Cup Airblast Fuel Injector Based on Backpropagation (BP) Neural Network Model.

This study was dedicated to introducing a new method for predicting the Sauter mean diameter (SMD) buildup in the swirl cup airblast fuel injector. There have been considerable difficulties with predicting SMD mainly because of complicated flow characteristics in a spray. Therefore, the backpropagation (BP) neural network-based machine learning was applied for the prediction of SMD as a function of geometry, condition parameters, and axial distance such as primary swirl number, secondary swirl number, venturi angle, mass flow rate of fuel, and relative air pressure. SMD was measured by a phase Doppler particle analyzer (PDPA). The results show that the prediction accuracy of the trained BP neural network was excellent with a coefficient of determination (R2) score of 0.9599, root mean square error (RMSE) score of 1.4613, and overall relative error within 20%. Through sensitivity analysis, the relative air pressure drop and primary swirl number were the largest and smallest factors affecting the value of SMD, respectively. Finally, the prediction accuracy of the BP neural network model is far greater than the current prediction correlations. Moreover, for the predicting target in the present study, the BP neural network shows the advantages of a simple structure and short running time compared with PSO-BP and GRNN. All these prove that the BP neural network is a novel and effective way to predict the SMD of droplets generated by a swirl cup airblast fuel injector.

Improvement of YX42° Cut LiTaO3 SAW Filters with Optical Proximity Effect Correction Method.

Due to the influence of the optical proximity effect (OPE), it is easy for a pattern of photoresistance to be inconsistent with a design pattern, thus damaging the performance of a SAW resonator. To solve this problem, this paper proposes an optimization method for SAW filters based on optical proximity correction (OPC). This method can avoid the tip discharge problem of SAW filters by suppressing the problem of rounding and shrinking of dummy electrode and electrode tail caused by OPE. This method increases the quality factor (Q) of the SAW resonator and thus decreases the insertion loss of the SAW filter. The filter increases the bandwidth by 1.8 MHz at -1.5 dB after applying the OPC method. Additionally, it improves the stability of the filter under high power conditions.

Chromatic visual evoked potentials identify optic nerve dysfunction in patients with Graves' orbitopathy.

PURPOSE: To explore visual dysfunction in Graves' orbitopathy (GO) objectively by analyzing chromatic visual evoked potentials (cVEP) and evaluate its diagnostic efficiency for dysthyroid optic neuropathy (DON). METHODS: In this cross-sectional study, we analyzed pattern-reversal VEP (pVEP), red-green (R-G) and blue-yellow (B-Y) cVEP in 93 subjects (21 with DON, Group A, 30 with GO, Group B, and 42 healthy controls, Group C) at Wuhan Union Hospital, China. RESULTS: Compared with Group C, the amplitudes of B-Y cVEP were significantly lower in Group B, whereas all amplitudes of cVEP, latencies and amplitudes of pVEP in Group A were significantly impaired. In addition, the pVEP latency at 60 arcmin (60'), pVEP amplitudes and R-G cVEP amplitudes were significantly different between Group A and B. Moreover, 60'cVEP R-G negative-positive (N-P) amplitude was correlated with crowding index (P = 0.001), the average thickness of ganglion cell layer and inner plexiform layer (P = 0.004). Furthermore, combination of 60'cVEP R-G amplitude and 60'pVEP P100 latency had better diagnostic efficiency than each single parameter, with optimal cut-off values of 14.20 µV and 110.65 ms, respectively. CONCLUSION: GO may induce electrophysiological changes. The presence of B-Y cVEP anomalies in moderate to severe GO patients may be an early sign of preclinical DON. A decline in 60'cVEP R-G amplitude is associated with apical crowding and thinner inner intra-retinal layers. The combination of 60'cVEP R-G N-P amplitude and 60'pVEP latency can be a useful diagnostic index for DON.

0D boron carbon nitride quantum dots decorated 2D Bi4O5I2 as 0D/2D efficient visible-light-driven photocatalysts for tetracyclines photodegradation.

A series of 0D boron carbon nitride quantum dots (BCNQDs) modified 2D Bi4O5I2 (0D/2D Bi4O5I2/BCNQDs) composites were synthesized and applied to photodegradation of tetracyclines (TCs), including tetracycline (TC) and oxytetracycline (OTC). The Bi4O5I2/BCNQDs (1) (1 mL BCNQDs) composite exhibits the highest photocatalytic performance for TCs degradation. The degradation rate constants of TC and OTC by the optimal sample were 4.95 and 2.17 times that of Bi4O5I2, respectively. This can be attributed to the fact that the narrow bandgap Bi4O5I2 is the electron acceptor, and the oxygen-containing functional group with the negative charge on BCNQDs can promote the formation of photoexcited holes, which makes the effective separation of photoexcited carriers easier. Furthermore, the active substance (h+ and ·O2-) is the major active substance for TCs photodegradation. On this basis, the possible photocatalytic reaction mechanism of Bi4O5I2/BCNQDs (1) composite is proposed. This study provides a new idea for 0D/2D interface engineering of BCNQDs heterojunction.

Secretions from hypochlorous acid-treated tumor cells delivered in a melittin hydrogel potentiate cancer immunotherapy.

Autologous tumor cells and cell-derived secretions (CDS) can induce antitumor immune responses. The conditions in which cells are cultured and treated impact CDS, and cellular insults alter their composition and function. In this study, we generated CDS from tumor cells exposed to normal culture conditions, hypoxia, cisplatin, radiotherapy, photodynamic therapy, or hypochlorous acid (HOCl). In vitro HOCl-CDS showed the strongest stimulatory effects on dendritic cells and macrophages compared to CDS generated by hypoxia, cisplatin, radiotherapy or photodynamic therapy. To improve HOCl-CDS activity at the tumor site, we loaded HOCl-CDS into a melittin-encapsulated hydrogel scaffold. When injected intratumorally, the HOCl-CDS hydrogel promoted tumor cell death, cytotoxic T lymphocyte infiltration, and tumor-associated macrophage reprogramming towards an M1 phenotype. The hydrogel inhibited tumor growth and prolonged the survival of mice bearing B16-F10 melanoma. Furthermore, hydrogel-delivered HOCl-CDS augmented the antitumor effects of immune checkpoint blockade. These results underscore the importance of the CDS generation method and delivery approach for improving cancer immunotherapy.

3'HS1 CTCF binding site in human ß-globin locus regulates fetal hemoglobin expression.

Mutations in the adult ß-globin gene can lead to a variety of hemoglobinopathies, including sickle cell disease and ß-thalassemia. An increase in fetal hemoglobin expression throughout adulthood, a condition named hereditary persistence of fetal hemoglobin (HPFH), has been found to ameliorate hemoglobinopathies. Deletional HPFH occurs through the excision of a significant portion of the 3' end of the ß-globin locus, including a CTCF binding site termed 3'HS1. Here, we show that the deletion of this CTCF site alone induces fetal hemoglobin expression in both adult CD34+ hematopoietic stem and progenitor cells and HUDEP-2 erythroid progenitor cells. This induction is driven by the ectopic access of a previously postulated distal enhancer located in the OR52A1 gene downstream of the locus, which can also be insulated by the inversion of the 3'HS1 CTCF site. This suggests that genetic editing of this binding site can have therapeutic implications to treat hemoglobinopathies.

GOLPH3 promotes glioma progression by enhancing PHB2-mediated autophagy.

Due to the hypoxia and nutrient deficiency microenvironment, malignant glioma exhibits high autophagy activity and autophagy plays a significant role in the occurrence and development of glioma. However, the potential molecular mechanism of autophagy in glioma remains unknown. In this study, we demonstrated that Golgi phosphorylation protein 3 (GOLPH3), a highly conserved protein basically concentrates in the trans-Golgi network, promoted glioma autophagy. Inhibiting autophagy by using chloroquine suppressed the stimulating effect of GOLPH3 on glioma malignant development both in vitro and in vivo. Mechanistically, GOLPH3 interacted with and recruited prohibitin-2 (PHB2), an autophagy receptor of mitochondrion, and LC3-II. PHB2 promoted cell autophagy and down-regulation of PHB2 abolished the effect of GOLPH3 on autophagy. On the side, the relative mRNA and protein levels of GOLPH3 and PHB2 were positively associated with each other and both also correlated with autophagy in glioma tissues. Together, our results revealed that GOLPH3 promotes glioma progression by enhancing PHB2-mediated autophagy and inhibiting autophagy may benefit glioma patients with GOLPH3 high level. The novel GOLPH3-PHB2-autophagy axis maybe a potential and prospective therapeutic target for gliomas.

Injectable Hydrogel as a Unique Platform for Antitumor Therapy Targeting Immunosuppressive Tumor Microenvironment.

Cancer immunotherapy can boost the immune response of patients to eliminate tumor cells and suppress tumor metastasis and recurrence. However, immunotherapy resistance and the occurrence of severe immune-related adverse effects are clinical challenges that remain to be addressed. The tumor microenvironment plays a crucial role in the therapeutic efficacy of cancer immunotherapy. Injectable hydrogels have emerged as powerful drug delivery platforms offering good biocompatibility and biodegradability, minimal invasion, convenient synthesis, versatility, high drug-loading capacity, controlled drug release, and low toxicity. In this review, we summarize the application of injectable hydrogels as a unique platform for targeting the immunosuppressive tumor microenvironment.

Large DNA Methylation Nadirs Anchor Chromatin Loops Maintaining Hematopoietic Stem Cell Identity.

Higher-order chromatin structure and DNA methylation are implicated in multiple developmental processes, but their relationship to cell state is unknown. Here, we find that large (>7.3 kb) DNA methylation nadirs (termed "grand canyons") can form long loops connecting anchor loci that may be dozens of megabases (Mb) apart, as well as inter-chromosomal links. The interacting loci cover a total of ∼3.5 Mb of the human genome. The strongest interactions are associated with repressive marks made by the Polycomb complex and are diminished upon EZH2 inhibitor treatment. The data are suggestive of the formation of these loops by interactions between repressive elements in the loci, forming a genomic subcompartment, rather than by cohesion/CTCF-mediated extrusion. Interestingly, unlike previously characterized subcompartments, these interactions are present only in particular cell types, such as stem and progenitor cells. Our work reveals that H3K27me3-marked large DNA methylation grand canyons represent a set of very-long-range loops associated with cellular identity.

GOLPH3 Regulates Exosome miRNA Secretion in Glioma Cells.

We aimed to examine whether golgi protein GOLPH3 could affect the secretion of glioma cell-derived exosomes. The exosomes were extracted by ultra-centrifugation from the supernatant of U251 and U87 cell cultures and identified by transmission electron microscopy (TEM), Malvern analyzer, and western blot. The quantity of exosomes was examined by measuring the total protein levels and the number of multiple vesicle bodies (MVBs), the source of exosomes. The exosome miRNAs were analyzed by high-throughput sequencing followed by GO and KEGG analysis, and validated by qRT-PCR. GOLPH3 could not affect the total protein levels of exosomes and the number of MVBs. However, we found 149 differentially expressed miRNAs in exosomes between vector and GOLPH3 over-expression group, and 14 miRNAs were only examined in GOLPH3 over-expression cells. The predicted target genes of these miRNAs had functions in binding and catalytic activity, which were enriched in the pathways of endocytosis, RNA transportation, thyroid hormone signaling and miRNAs in cancer. GOLPH3 could not affect the quantity of exosomes, but rather contribute to miRNA expression in exosomes, which may play some functions in the promotion effect of GOLPH3 on glioma development.

Whole genome shotgun sequence of Bacillus amyloliquefaciens TF28, a biocontrol entophytic bacterium.

Bacillus amyloliquefaciens TF28 is a biocontrol endophytic bacterium that is capable of inhibition of a broad range of plant pathogenic fungi. The strain has the potential to be developed into a biocontrol agent for use in agriculture. Here we report the whole-genome shotgun sequence of the strain. The genome size of B. amyloliquefaciens TF28 is 3,987,635 bp which consists of 3754 protein-coding genes, 65 tandem repeat sequences, 47 minisatellite DNA, 2 microsatellite DNA, 63 tRNA, 7rRNA, 6 sRNA, 3 prophage and CRISPR domains.