Ectosomal PKM2 Promotes HCC by Inducing Macrophage Differentiation and Remodeling the Tumor Microenvironment.

Tumor-derived extracellular vesicles are important mediators of cell-to-cell communication during tumorigenesis. Here, we demonstrated that hepatocellular carcinoma (HCC)-derived ectosomes remodel the tumor microenvironment to facilitate HCC progression in an ectosomal PKM2-dependent manner. HCC-derived ectosomal PKM2 induced not only metabolic reprogramming in monocytes but also STAT3 phosphorylation in the nucleus to upregulate differentiation-associated transcription factors, leading to monocyte-to-macrophage differentiation and tumor microenvironment remodeling. In HCC cells, sumoylation of PKM2 induced its plasma membrane targeting and subsequent ectosomal excretion via interactions with ARRDC1. The PKM2-ARRDC1 association in HCC was reinforced by macrophage-secreted cytokines/chemokines in a CCL1-CCR8 axis-dependent manner, further facilitating PKM2 excretion from HCC cells to form a feedforward regulatory loop for tumorigenesis. In the clinic, ectosomal PKM2 was clearly detected in the plasma of HCC patients. This study highlights a mechanism by which ectosomal PKM2 remodels the tumor microenvironment and reveals ectosomal PKM2 as a potential diagnostic marker for HCC.

Phage-assisted evolution of compact Cas9 variants targeting a simple NNG PAM.

Compact Cas9 nucleases hold great promise for therapeutic applications. Although several compact Cas9 nucleases have been developed, many genomic loci still could not be edited due to a lack of protospacer adjacent motifs (PAMs). We previously developed a compact SlugCas9 recognizing an NNGG PAM. Here we demonstrate that SlugCas9 displays comparable activity to SpCas9. We developed a simple phage-assisted evolution to engineer SlugCas9 for unique PAM requirements. Interestingly, we generated a SlugCas9 variant (SlugCas9-NNG) that could recognize an NNG PAM, expanding the targeting scope. We further developed a SlugCas9-NNG-based adenine base editor and demonstrated that it could be delivered by a single adeno-associated virus to disrupt PCSK9 splice donor and splice acceptor. These genome editors greatly enhance our ability for in vivo genome editing.

Identification of SaCas9 orthologs containing a conserved serine residue that determines simple NNGG PAM recognition.

Due to different nucleotide preferences at target sites, no single Cas9 is capable of editing all sequences. Thus, this highlights the need to establish a Cas9 repertoire covering all sequences for efficient genome editing. Cas9s with simple protospacer adjacent motif (PAM) requirements are particularly attractive to allow for a wide range of genome editing, but identification of such Cas9s from thousands of Cas9s in the public database is a challenge. We previously identified PAMs for 16 SaCas9 orthologs. Here, we compared the PAM-interacting (PI) domains in these orthologs and found that the serine residue corresponding to SaCas9 N986 was associated with the simple NNGG PAM requirement. Based on this discovery, we identified five additional SaCas9 orthologs that recognize the NNGG PAM. We further identified three amino acids that determined the NNGG PAM requirement of SaCas9. Finally, we engineered Sha2Cas9 and SpeCas9 to generate high-fidelity versions of Cas9s. Importantly, these natural and engineered Cas9s displayed high activities and distinct nucleotide preferences. Our study offers a new perspective to identify SaCas9 orthologs with NNGG PAM requirements, expanding the Cas9 repertoire.

Exploring NK-Cell molecules that impact the immune response and microenvironment in head and neck squamous cell carcinoma.

NK cells play a role in various cancers, but their role in head and neck squamous cell carcinoma (HNSCC) still needs to be explored. All public data are obtained from the Cancer Genome Atlas Program (TCGA) database. All analysis was performed using specific packages in R software. In our study, we quantified the immune microenvironment of HNSCC through multiple algorithms. Next, we identified NK cell-associated genes by quantifying NK cells, including SSNA1, TRIR, PAXX, DPP7, WDR34, EZR, PHLDA1 and ELOVL1. Then, we explored the single-cell expression pattern of these genes in the HNSCC microenvironment. Univariate Cox regression analysis indicated that the EZR, PHLDA1 and ELOVL1 were related to the prognosis of HNSCC patients. Following this, we selected EZR for further analysis. Our results showed that the patients with high EZR expression might have a poor prognosis and worse clinical features. Biological enrichment analysis showed that EZR is associated with many oncogenic pathways and a higher tumour stemness index. Meanwhile, we found that EZR can remodel the immune microenvironment of HNSCC. Moreover, we noticed that EZR could affect the immunotherapy and specific drug sensitivity, making it an underlying clinical target. In summary, our results can improve the understanding of NK cell in HNSCC. Meanwhile, we identified EZR as the underlying clinical target of HNSCC.

Engineering Dispersive-Dissipative Modal Coupling in Hybrid Optical Microresonators.

Hybrid microresonators have served an intriguing platform for fundamental research and applied photonics. Here, we study the plasmonics-engineered coupling between degenerate optical whispering gallery modes, which can be tuned in a complex space featuring the dissipative strong, dispersive strong, and weak coupling regimes. Experimentally, the engineering of a single plasmonic resonance to a cavity mode family is examined in a waveguide-integrated high-Q microdisk, from which the complex coupling coefficients are extracted and agree well with theoretical predictions. The coupling strength over 10 GHz is achieved for both dissipative and dispersive interactions, showing a remarkable enhancement compared to that induced by a dielectric scatterer. Furthermore, the far fields of hybridized cavity modes are measured, revealing the coherent interference between the radiative channels. Our results shed light on the engineering of whispering gallery modes through plasmonic resonances, and provide fundamental guidance to practical microcavity devices.

Genome editing with natural and engineered CjCas9 orthologs.

CjCas9 is one of the smallest CRISPR-associated (Cas9) nucleases for mammalian genome editing. However, it requires a long N4RYAC (R = A or G; Y = C or T) protospacer-adjacent motif (PAM), limiting its DNA targeting scope. In this study, we investigated the PAMs of three CjCas9 orthologs, including Hsp1Cas9, Hsp2Cas9, and CcuCas9, by performing a GFP-activation assay. Interestingly, Hsp1Cas9 and CcuCas9 recognized unique N4RAA and N4CNA PAMs, respectively. We further generated an Hsp1Cas9-Hsp2Cas9 chimeric Cas9 (Hsp1-Hsp2Cas9), which recognized a simple N4CY PAM. Genome-wide off-target analysis revealed that Hsp1-Hsp2Cas9 has very few off-targets compared to SpCas9. By analyzing the crystal structure of CjCas9, we identified eight mutations that can improve the specificity and generate a high-fidelity Hsp1-Hsp2Cas9-Y. Hsp1-Hsp2Cas9-Y enables the knockout of B4GALNT2 and CMAH in porcine fetal fibroblasts (PFFs). Moreover, we developed a high-fidelity Hsp1-Hsp2Cas9-KY which displayed undetectable off-targets revealed by GUIDE-seq at four tested loci. These natural and engineered Cas9 nucleases enabled efficient genome editing in multiple mammalian cells, expanding the DNA targeting scope.

Single-mode characteristic of a supermode microcavity Raman laser.

Microlasers in near-degenerate supermodes lay the cornerstone for studies of non-Hermitian physics, novel light sources, and advanced sensors. Recent experiments of the stimulated scattering in supermode microcavities reported beating phenomena, interpreted as dual-mode lasing, which, however, contradicts their single-mode nature due to the clamped pump field. Here, we investigate the supermode Raman laser in a whispering-gallery microcavity and demonstrate experimentally its single-mode lasing behavior with a side-mode suppression ratio (SMSR) up to 37 dB, despite the emergence of near-degenerate supermodes by the backscattering between counterpropagating waves. Moreover, the beating signal is recognized as the transient interference during the switching process between the two supermode lasers. Self-injection is exploited to manipulate the lasing supermodes, where the SMSR is further improved by 15 dB and the laser linewidth is below 100 Hz.

[Gualou Xiebai Banxia Decoction treats type 2 diabetes mellitus combined with acute myocardial infarction via chemerin/ CMKLR1/PPARα signaling pathway].

This study aims to investigate the effects of Gualou Xiebai Banxia Decoction(GXBD) on type 2 diabetes mellitus(T2DM) combined with acute myocardial infarction(AMI) in rats via chemerin/chemokine-like receptor 1(CMKLR1)/peroxisome proliferator-activated receptor α(PPARα) signaling pathway, and to explore the mechanism of GXBD in alleviating glucose and lipid metabolism disorders. The SD rats were randomized into control, model, positive control, and low-and high-dose GXBD groups. The rat model of T2DM was established by administration with high-fat emulsion(HFE) by gavage and intraperitoneal injection with streptozotocin, and then coronary artery ligation was performed to induce AMI. The control and model groups were administrated with the equal volume of normal saline, and other groups were administrated with corresponding drugs by gavage. Changes in relevant metabolic indicators were assessed by ELISA and biochemical assays, and the protein levels of chemerin, CMKLR1, and PPARα in the liver, abdominal fat, and heart were determined by Western blot. The results showed that GXBD alleviated the myocardial damage and reduced the levels of blood lipids, myocardial enzymes, and inflammatory cytokines, while it did not lead to significant changes in blood glucose. Compared with the model group, GXBD down-regulated the expression of chemerin in peripheral blood and up-regulated the expression of cyclic adenosine monophosphate(cAMP) and protein kinase A(PKA) in the liver. After treatment with GXBD, the protein levels of chemerin and CMKLR1 in the liver, abdominal fat, and heart were down-regulated, while the protein levels of PPARα in the liver and abdominal fat were up-regulated. In conclusion, GXBD significantly ameliorated the disorders of glycolipid metabolism in the T2DM-AMI model by regulating the chemerin/CMKLR1/PPARα signaling pathway to exert a protective effect on the damaged myocardium. This study provides a theoretical basis for further clinical study of GXBD against T2DM-AMI and is a manifestation of TCM treatment of phlegm and turbidity causing obstruction at the protein level.

The Ordered Mesoporous Barium Ferrite Compounded with Nitrogen-Doped Reduced Graphene Oxide for Microwave Absorption Materials.

Nanocomposites with hierarchical pore structure hold great potentials for applications in the field of microwave-absorbing materials because of their lightweight and high-efficiency absorption properties. Herein, M-type barium ferrite (BaM) with ordered mesoporous structure (M-BaM) is prepared via a sol-gel process enhanced by mixed anionic and cationic surfactants. The surface area of M-BaM is enhanced almost ten times compared with BaM together with 40% reflection loss enhancing. Then M-BaM compounded with nitrogen-doped reduced graphene oxide (MBG) is synthesized via hydrothermal reaction in which the reduction and nitrogen doping of graphene oxide (GO) in situ occur simultaneously. Interestingly, the mesoporous structure is able to provide opportunity for reductant to enter the bulk M-BaM reducing its Fe3+ to Fe2+ and further forms Fe3 O4 . It requires an optimal balance among the remained mesopores in MBG, formed Fe3 O4 , and CN in nitrogen-doped graphene (N-RGO) for optimizing impedance matching and greatly increasing multiple reflections/interfacial polarization. MBG-2 (GO:M-BaM = 1:10) achieves the minimum reflection loss of -62.6 dB with an effective bandwidth of 4.2 GHz at an ultra-thin thickness of 1.4 mm. In addition, the marriage of mesoporous structure of M-BaM and light mass of graphene reduces the density of MBG.

Efficient Correction of a Hypertrophic Cardiomyopathy Mutation by ABEmax-NG.

[Figure: see text].

Influence of Environmental Factors on Salivary Microbiota and Their Metabolic Pathway: Next-Generation Sequencing Approach.

The current study aimed to investigate the effect of periodontitis and long-term heavy metal (HM) exposure on the salivary microbiome. The patients were divided into four groups as Wu Wei control (WWC) group involved healthy individuals, Wu Wei periodontitis (WWP) patients having periodontitis, Jing Chang with metal pollution periodontally healthy individuals (JCP), and Kuang periodontitis (KP). The most abundant bacteria identified at the phylum level in the WWC group were Bacteroides, Firmicutes, and Fusobacteria. Firmicutes were observed in a significantly higher proportion in the KP group than in the WWC, WWP, and JCP. At the genus level, the WWC has major dominating bacterial genera (such as Leptotrichia, Neisseria, and Fusobacterium) which were similar to WWP and KP group. The significant difference (p < 0.05) was found in alpha diversity while in beta diversity, the significant (p = 0.005) results were found among the four groups. The correlation of oral microbiota revealed that HMs present in the soil (Cr, Ni, and Cu) are associated with the growth of Capnocytophaga, Selenomonas, Aggregatibacter, and Campylobacter. The bacterial functions in the KP group were higher in translation and nucleotide metabolism than in the WWP group. This demonstrated that long-term exposure to HMs can influence the salivary microbiota which can alter the functioning, and diversity of bacteria.

Discovery and engineering of small SlugCas9 with broad targeting range and high specificity and activity.

The compact CRISPR/Cas9 system, which can be delivered with their gRNA and a full-length promoter for expression by a single adeno-associated virus (AAV), is a promising platform for therapeutic applications. We previously identified a compact SauriCas9 that displays high activity and requires a simple NNGG PAM, but the specificity is moderate. Here, we identified three compact Cas9 orthologs, Staphylococcus lugdunensis Cas9 (SlugCas9), Staphylococcus lutrae Cas9 (SlutrCas9) and Staphylococcus haemolyticus Cas9 (ShaCas9), for mammalian genome editing. Of these three Cas9 orthologs, SlugCas9 recognizes a simple NNGG PAM and displays comparable activity to SaCas9. Importantly, we generated a SlugCas9-SaCas9 chimeric nuclease, which has both high specificity and high activity. We finally engineered SlugCas9 with mutations to generate a high-fidelity variant that maintains high specificity without compromising on-target editing efficiency. Our study offers important minimal Cas9 tools that are ideal for both basic research and clinical applications.

Greenhouse gas emissions and peak trend of commercial vehicles in China.

Commercial vehicles are important within the context of global warming, since they exhibit greenhouse gas (GHG) emissions that are disproportionate to their quantity. The aim of this study was to create a bottom-up GHG emissions assessment model which considers GHG emissions of newly produced commercial vehicles and those in current use. Through this study, the number of future commercial vehicles were predicted, thereby facilitating a simulation of future GHG emissions. Our results show that the total GHG emissions of commercial vehicles in 2019 was 580 million t CO2-eq.. Among them, the GHG emissions stemming from the production of new commercial vehicles accounted for ∼0.3% of the emissions, whereas the use stage accounted for more than 99.0%. Moreover, the future ownership of commercial vehicles depends on GDP and the demand of freight and passenger transport. The ownership of commercial vehicles was predicted about 36.61 million in 2025, 45.44 million in 2030 and 55.85 million in 2035. The carbon peak of commercial vehicles varies across different scenarios, peaking around 2031-2034, at 680-780 million t CO2-eq.. This study systematically simulated the carbon peak of commercial vehicles, contributing toward a deeper understanding of commercial vehicles within the context of GHG emissions. These results can be applied toward creating quantitatively-driven pathways for carbon peak or neutrality targets in the commercial vehicle sector.

High-value utilisation of PGM-containing residual oil: Recovery of inorganic acids, potassium, and PGMs using a zero-waste approach.

Residual oil containing platinum group metals (PGMs), which is under-researched, can easily pose resource waste and environmental risks. PGMs feature as scarce strategic metals, and inorganic acids and potassium salts are also considered valuable. An integrated process for the harmless treatment and recovery of useful resources from residual oil is proposed herein. This work developed a zero-waste process based on the study of the main components and characteristics of the PGM-containing residual oil. The process consists of three modules: pre-treatment for phase separation, liquid-phase resource utilisation, and solid-phase resource utilisation. Separating the residual oil into liquid and solid phases allows for the maximum recovery of valuable components. However, concerns about the accurate determination of valued components emerged. Findings revealed that Fe and Ni are highly susceptible to spectral interference in the PGMs test when using the inductively coupled plasma method. After studying 26 PGM emission lines, Ir 212.681 nm, Pd 342.124 nm, Pt 299.797 nm, and Rh 343.489 nm were reliably identified. Finally, formic acid (81.5 g/t), acetic acid (117.2 kg/t), propionic acid (291.9 kg/t), butyric acid (3.6 kg/t), potassium salt (553.3 kg/t), Ir (27.8 g/t), Pd (10960.0 g/t), Pt (193.1 g/t), and Rh (109.8 g/t) were successfully obtained from the PGM-containing residual oil. This study provides a helpful reference for the determination of PGM concentrations and high-value utilisation of PGM-containing residual oil.

Vibrational Kerr Solitons in an Optomechanical Microresonator.

Kerr soliton microcombs in microresonators have been a prominent miniaturized coherent light source. Here, for the first time, we demonstrate the existence of Kerr solitons in an optomechanical microresonator, for which a nonlinear model is built by incorporating a single mechanical mode and multiple optical modes. Interestingly, an exotic vibrational Kerr soliton state is found, which is modulated by a self-sustained mechanical oscillation. Besides, the soliton provides extra mechanical gain through the optical spring effect, and results in phonon lasing with a red-detuned pump. Various nonlinear dynamics is also observed, including limit cycle, higher periodicity, and transient chaos. This work provides a guidance for not only exploring many-body nonlinear interactions, but also promoting precision measurements by featuring superiority of both frequency combs and optomechanics.

Regulated Photon Transport in Chaotic Microcavities by Tailoring Phase Space.

Manipulating light dynamics in optical microcavities has been made mainly either in real or momentum space. Here we report a phase-space tailoring scheme, simultaneously incorporating spatial and momentum dimensions, to enable deterministic and in situ regulation of photon transport in a chaotic microcavity. In the time domain, the chaotic photon transport to the leaky region can be suppressed, and the cavity resonant modes show stronger temporal confinement with quality factors being improved by more than 1 order of magnitude. In the spatial domain, the emission direction of the cavity field is controlled on demand through rerouting chaotic photons to a desired channel, which is verified experimentally by the far-field pattern of a quantum-dot microlaser. This work paves a way to in situ study of chaotic physics and promoting advanced applications such as arbitrary light routing, ultrafast random bit generation, and multifunctional on-chip lasers.

KeMRE: Knowledge-enhanced medical relation extraction for Chinese medicine instructions.

Medicine instructions usually contain rich medical relations, and extracting them is very helpful for many downstream tasks such as medicine knowledge graph construction and medicine side-effect prediction. Existing relation extraction (RE) methods usually predict relations between entities from their contexts and do not consider medical knowledge. However, understanding a part of medical relations may need some expert knowledge in the medical field, making it challenging for existing methods to achieve satisfying performances of medical RE. In this paper, we propose a knowledge-enhanced framework for medical RE, which can exploit medical knowledge of medicines to better conduct medical RE on Chinese medicine instructions. We first propose a BERT-CNN-LSTM based framework for text modeling and learn representations of characters from their contexts. Then we learn representations of each entity by aggregating representations of their characters. Besides, we propose a CNN-LSTM based framework for entity modeling and learn entity representations from their relatedness. In addition, there are usually many different instructions for the same medicine, which usually share general knowledge on this medicine. Thus, to obtain medical knowledge of medicines, we annotate relations on a randomly-sampled instruction of each medicine. Then we build knowledge embeddings to represent potential relations between entities from knowledge of medicines. Finally, we use an MLP network to predict relations between entities from their representations and knowledge embeddings. Extensive experiments on a real-world dataset show that our method can significantly outperform existing methods.

Culture and purification of SD rat corpus cavernosum endothelial cells by enzymatic digestion combined with mechanical extrusion and fixed-point digestion.

To explore a new method of in vitro culture and purification of rat corpus cavernosum endothelial cells (CCECs). Male Sprague-Dawley rats' penile tissue were digested with elastase or collagenase combined with mechanical extrusion to isolate and culture the CCECs. The fixed-point digestion method was used to purify the primary cells. High-purity CCECs were successfully isolated. Following the digestion of the primary CCECs by elastase or collagenase coupled with mechanical extrusion, the cells were paving stone- and cobblestone-shaped over 10 days. The cell purity yielded in the second generation (P2) CCECs after using the fixed-point digestion method was significantly high. Compared with primary CCECs extracted by elastase digestion combined with the mechanical extrusion method, CCECs cultured by collagenase digestion yielded higher purity and a more stable morphology after fixed-point digestion and purification. Immunofluorescence staining of the third generation CCECs and the expression results of endothelial cell-associated marker antibodies CD31 and VWF were positive, and flow cytometry showed the purity of CCECs was 96.9%. Enzymatic digestion combined with mechanical extrusion and fixed-point digestion is a simple, economical method for in vitro culture and purification of CCECs, which is conducive to studying the pathophysiological mechanisms of endothelial dysfunction and erectile dysfunction.

Real-Time Learning and Recognition of Assembly Activities Based on Virtual Reality Demonstration.

Teaching robots to learn through human demonstrations is a natural and direct method, and virtual reality technology is an effective way to achieve fast and realistic demonstrations. In this paper, we construct a virtual reality demonstration system that uses virtual reality equipment for assembly activities demonstration, and using the motion data of the virtual demonstration system, the human demonstration is deduced into an activity sequence that can be performed by the robot. Through experimentation, the virtual reality demonstration system in this paper can achieve a 95% correct rate of activity recognition. We also created a simulated ur5 robotic arm grasping system to reproduce the inferred activity sequence.