ACKR1+ Endothelial Cells Mediate Leukocyte Infiltration and Synergize with SFRP2/ASPN+ Fibroblasts to Promote Skin Fibrosis in Systemic Sclerosis.

BACKGROUND: Skin fibrosis is the most typical pathological manifestation of systemic sclerosis (SSc) and localized scleroderma (LS) with unclear etiology and few effective treatments. Though excessive collagen secretion by fibroblasts is the primary cause of skin fibrosis, many lines of evidence suggested that vascular damage was the initiating event and various cell types along with fibroblasts worked together to contribute to the pathogenesis of skin fibrosis. OBJECTIVES: We sought to explore the relationships between vascular endothelial cell lesions and immune cell infiltration, along with the cell-cell interactions among various cell types within the fibrotic skin ecosystem. METHODS: Single-cell RNA-seq (10x Genomics) was performed on skin biopsies of 3 healthy donors and 7 SSc patients in Chinese. The additional 3 localized scleroderma patients' data from NCBI database (GSE160536) were integrated by Harmony. CellChat package (v1.5.0) was applied to analyze cell communication network. Transwell assay and subcutaneous bleomycin (BLM) injection in mice were used to explore the role of ACKR1 on immune cell infiltration. Milo single-cell western blot was applied to show the activation of fibroblast subclusters. RESULTS: A total of 62,295 cells were obtained and subpopulations of stromal and immune cells were identified. Interaction network analysis revealed that multiple chemokines secreted by macrophages, pericytes, and pro-inflammatory fibroblasts could bind with Duffy antigen/receptor for chemokines (ACKR1), which is highly expressed on ACKR1+ endothelial cells of lesion skin. Transwell assay revealed that over-expressed ACKR1 in HUVEC facilitated leukocyte infiltration under the treatment of IL8. The BLM mice showed enhanced ACKR1 expression, massive immune cell infiltration, and fibrosis in skin, which could be attenuated by ACKR1 inhibition. Furthermore, infiltrated macrophages with TGFB1 or PDGFB high production could activate SFRP2/ASPN+ fibroblasts to contribute to excessive accumulation of extracellular matrix (ECM), and the SOX4-ASPN axis plays an important role in the TGF-ß signaling cascade and the etiology of skin fibrosis. CONCLUSIONS: Our results reveal that highly expressed ACKR1 in endothelial cells of fibrotic skin tissue promotes immune cell infiltration, and SFRP2/ASPN+ fibroblasts synergize to exacerbate skin fibrosis.

Establishing the relationship between subjective perception and neural responses: Insights from correlation analysis and representational similarity analysis.

Exploring the relationship between sensory perception and brain responses holds important theoretical and clinical implications. However, commonly used methodologies like correlation analysis performed either intra- or inter- individually often yield inconsistent results across studies, limiting their generalizability. Representational similarity analysis (RSA), a method that assesses the perception-response relationship by calculating the correlation between behavioral and neural patterns, may offer a fresh perspective to reveal novel findings. Here, we delivered a series of graded sensory stimuli of four modalities (i.e., nociceptive somatosensory, non-nociceptive somatosensory, visual, and auditory) to/near the left or right hand of 107 healthy subjects and collected their single-trial perceptual ratings and electroencephalographic (EEG) responses. We examined the relationship between sensory perception and brain responses using within- and between-subject correlation analysis and RSA, and assessed their stability across different numbers of subjects and trials. We found that within-subject and between-subject correlations yielded distinct results: within-subject correlation revealed strong and reliable correlations between perceptual ratings and most brain responses, while between-subject correlation showed weak correlations that were vulnerable to the change of subject number. In addition to verifying the correlation results, RSA revealed some novel findings, i.e., correlations between behavioral and neural patterns were observed in some additional neural responses, such as "γ-ERS" in the visual modality. RSA results were sensitive to the trial number, but not to the subject number, suggesting that consistent results could be obtained for studies with relatively small sample sizes. In conclusion, our study provides a novel perspective on establishing the relationship between behavior and brain activity, emphasizing that RSA holds promise as a method for exploring this pattern relationship in future research.

A universal design of restructured dimer antigens: Development of a superior vaccine against the paramyxovirus in transgenic rice.

The development of vaccines, which induce effective immune responses while ensuring safety and affordability, remains a substantial challenge. In this study, we proposed a vaccine model of a restructured "head-to-tail" dimer to efficiently stimulate B cell response. We also demonstrate the feasibility of using this model to develop a paramyxovirus vaccine through a low-cost rice endosperm expression system. Crystal structure and small-angle X-ray scattering data showed that the restructured hemagglutinin-neuraminidase (HN) formed tetramers with fully exposed quadruple receptor binding domains and neutralizing epitopes. In comparison with the original HN antigen and three traditional commercial whole virus vaccines, the restructured HN facilitated critical epitope exposure and initiated a faster and more potent immune response. Two-dose immunization with 0.5 µg of the restructured antigen (equivalent to one-127th of a rice grain) and one-dose with 5 µg completely protected chickens against a lethal challenge of the virus. These results demonstrate that the restructured HN from transgenic rice seeds is safe, effective, low-dose useful, and inexpensive. We provide a plant platform and a simple restructured model for highly effective vaccine development.

Enhancing pain modulation: the efficacy of synchronous combination of virtual reality and transcutaneous electrical nerve stimulation.

Introduction: Virtual reality (VR) and transcutaneous electrical nerve stimulation (TENS) have emerged as effective interventions for pain reduction. However, their standalone applications often yield limited analgesic effects, particularly in certain painful conditions. Aims: Our hypothesis was that the combination of VR with TENS in a synchronous manner could produce the best analgesic effect among the four experimental conditions. Methods: To address this challenge, we proposed a novel pain modulation strategy that synchronously combines VR and TENS, aiming to capitalise on both techniques' complementary pain modulation mechanisms. Thirty-two healthy subjects participated in the study and underwent three types of interventions: VR alone, a combination of VR with conventional TENS, and a combination of VR with synchronous TENS. Additionally, a control condition with no intervention was included. Perceived pain intensity, pain unpleasantness, positive and negative affect scores, and electroencephalographic (EEG) data were collected before and after the interventions. To delve into the potential moderating role of pain intensity on the analgesic efficacy of VR combined with synchronous TENS, we incorporated two distinct levels of painful stimuli: one representing mild to moderate pain (ie, low pain) and the other representing moderate to severe pain (ie, high pain). Results: Our findings revealed that both combination interventions exhibited superior analgesic effects compared with the VR-alone intervention when exposed to low and high pain stimuli. Notably, the combination of VR with synchronous TENS demonstrated greater analgesic efficacy than the combination of VR with conventional TENS. EEG data further supported these results, indicating that both combination interventions elicited a greater reduction in event-related potential magnitude compared with the VR-alone intervention during exposure to low and high pain stimuli. Moreover, the synchronous combination intervention induced a more significant reduction in N2 amplitude than the VR-alone intervention during exposure to low pain stimuli. No significant differences in EEG response changes were detected between the two combination interventions. Both combination interventions resulted in a greater reduction in negative affect compared with the VR-alone intervention. Conclusions: Altogether, our study highlights the effectiveness of the synchronous combination of VR and TENS in enhancing pain modulation. These findings offer valuable insights for developing innovative pain treatments, emphasising the importance of tailored and multifaceted therapeutic approaches for various painful conditions.

EGCG promotes the sensory function recovery in rats after dorsal root crush injury by upregulating KAT6A and inhibiting pyroptosis.

Dorsal root injury usually leads to irreversible sensory function loss and lacks effective treatments. (-)-epigallocatechin-3-gallate (EGCG) is reported to exert neuroprotective roles in the nervous systems. However, the function of EGCG in treating dorsal root injury remains unclear. Hence, we built the dorsal root crush injury (DRCI) rat model to be treated with EGCG, followed by the western blot, Enzyme-linked immunosorbent assay, and sensory behavior tests. We observed that EGCG can upregulate the Lysine acetyltransferase 6A (KAT6A) level and inhibit the pyroptosis, indicated by downregulated gasdermin-D, caspase-1, and interleukin 18 protein levels, and alleviate the neuropathic pain, indicated by the decreased paw withdraw threshold in Plantar test and decreased paw withdraw latency in von Frey test, and downregulated calcitonin gene-related peptide, nerve growth factor, and c-Fos protein levels. But EGCG cannot alleviate the neuropathic pain when the KAT6A was inhibited by CTX-0124143 and pyroptosis was activated by Miltirone. These combined results indicated that EGCG can promote the sensory function recovery in rats after DRCI via upregulating KAT6A and inhibiting pyroptosis, laying the foundation for EGCG to be a novel candidate for the treatment of dorsal root injury.

A multi-parameter tunable plasmon modulator.

Multi-parameter control of light is a key functionality to modulate optical signals in photonic integrated circuits for various applications. However, the traditional optical modulators can only control one or two properties of light at the same time. Herein, we propose a hybrid structure which can modulate the amplitude, wavelength and phase of surface plasmon polaritons (SPPs) simultaneously to overcome these limitations. The numerical results show that when the Fermi level of graphene changes from 0.3 to 0.9 eV, the variation of optical transmission, wavelength and phase are 32.7 dB, 428 nm and 306°, respectively. The demonstrated structure triggers an approach for the realization of ultracompact modulation and has potential applications in the fields of optical switches, communications and photo-detection.

Two patients with KDM3B variants and new presentations of Diets-Jongmans syndrome.

KDM3B is located on chromosome 5q31 and encodes KDM3B, which is involved in histone demethylation and epigenetic regulation. Pathogenic KDM3B variants cause a dominantly inherited disorder presenting with intellectual disability (ID), short stature, and facial dysmorphism, named Diets-Jongmans syndrome. We describe two patients with KDM3B variants presenting with Diets-Jongmans syndrome. Genetic testing was performed because of the clinical data and a lack of a clear diagnosis in both patients. Candidate variants were verified by Sanger sequencing. After KDM3B variants were detected, in silico tools were used to predict the pathogenicity of the missense variants. A minigene assay was performed to evaluate the splicing effects of the c.5070 + 1G > A variant on KDM3B. Patient 1 mainly presented with repetitive upper respiratory tract infection and patient 2 presented with palpitation, shortness of breath, and pitting edema; both had ID. Whole exome sequencing identified variants of KDM3B. Patient 1 had the de novo KDM3B c.5070 + 1G > A variant, whereas patient 2 had the c.2828G > A (p.R943Q) variant. Transcriptional experiments of the splicing variant c.5070 + 1G > A revealed aberrant transcripts leading to truncated protein products. We found two pathogenic variants in KDM3B, one of which is novel. Both patients had additional clinical presentations, and patient 1 had transient neutropenia. KDM3B c.5070 + 1G > A is the first KDM3B splice-site variant and was identified as a germline variant. Neutropenia and cardiomyopathy are newly found presentations of Diets-Jongmans syndrome. Our report enriches our knowledge of the genotypic spectrum of the KDM3B variants and phenotypic diversity of Diets-Jongmans syndrome.

[Progress of research on the genetic diseases caused by variants of mitochondrial aminoacyl-tRNA synthase gene].

As conserved enzymes with important functions, aminoacyl-tRNA synthetase are expressed ubiquitously in cells. These include cytoplasmic aminoacyl-tRNA synthetase, mitochondrial aminoacyl-tRNA synthetase and bifunctional aminoacyl-tRNA synthetase. Mitochondrial aminoacyl-tRNA synthetases catalyze the binding of amino acids with its corresponding tRNA in the mitochondria and participate in the translation of 13 subunits of oxidative phosphorylation enzyme complexes encoded by the mitochondrial genome. Mutations in genes encoding mitochondrial aminoacyl-tRNA synthase may cause a variety of genetic disorders. This review has summarized the clinical characteristics, molecular pathogenesis and treatment of genetic diseases caused by mutations of such genes.

Compact plasmon modulator with a high extinction ratio.

To keep pace with the demands in optical communications, electro-optic modulators should feature a high extinction ratio, offer a small footprint, and allow for practical detection. Herein, we demonstrate a compact plasmon modulator with a high extinction ratio where a compact modulation region composed of indium tin oxide (ITO) is embedded to the arms of the Mach-Zehnder (M-Z) interferometer. The modulator has a footprint of 20µm×12µm with a modulation region of 4µm×0.5µm. The numerical results show that the extinction ratio is 15.2 dB when the electron concentration of ITO is changed 4×1020cm-3. This type of modulator paves the way for future compact optoelectronic integration and has potential application in the fields of optical communication, photodetection, and sensing.

Study of novel NARS2 variants in patient of combined oxidative phosphorylation deficiency 24.

Background: NARS2 catalyzes the attachment of asparagine amino acids to mitochondrial tRNAAsn and is critical for efficient mitochondrial protein synthesis. Biallelic variants in NARS2 are associated with combined oxidative phosphorylation deficiency 24 (COXPD24) and autosomal recessive deafness-94. Methods: Patient information was obtained after recruitment. Genetic tests were performed using whole exome sequencing (WES) and Sanger sequencing. Structure prediction was based on the RaptorX and SWISS-MODEL platforms. The mRNA analysis of paternal variant was performed. Expression levels and dimerization of wild-type (WT) and mutant NARS2 were detected in human embryonic kidney (HEK) 293T cells. Mitochondrial localization of NARS2 variants was determined using immunofluorescence staining. Results: The patient presented early onset generalized epilepsy, myoclonic seizures, severe bilateral hearing impairment and affected liver and heart. WES identified two compound heterozygous variants in NARS2: c.1141A>G/p.Asn381Asp and c.1290G>C/p.Trp430Cys. In silico analysis predicted that both variants would cause significant and pathogenic changes in secondary structure. NARS2 c.1290G>C is a variant at the first nucleotide of an exon, a location thought to affect mRNA splicing. Although transcriptional experiments did not identify aberrant splicing, we observed a lower proportion of transcripts from the NARS2 c.1290G>C variant. An in vitro experiment showed that both variants impaired NARS2 expression, while mitochondrial localization and dimerization remained unaffected. Conclusions: The patient was diagnosed with COXPD24 caused by novel NARS2 variations. The cardiac dysfunction is identified for the first time. In vitro study revealed impairment of variants on NARS2 expression. These data enrich our knowledge regarding the phenotypic and genotypic spectra of NARS2.

Novel RARS2 Variants: Updating the Diagnosis and Pathogenesis of Pontocerebellar Hypoplasia Type 6.

BACKGROUND: Pontocerebellar hypoplasia type 6 (PCH6) is an early-onset encephalopathy with/without mitochondrial respiratory complex defects caused by recessive mutations in mitochondrial arginyl-tRNA synthetase (RARS2). Highly heterogeneous clinical phenotypes and numerous missense variations of uncertain significance make diagnosis difficult. Pathogenesis of PCH6 remains unclear. METHODS: Facial characteristics of patients were assessed. Genetic tests were performed. Structure prediction was based on the template from AlphaFold Protein Structure Database. Expression of mutant RARS2 was tested in HEK293T cells. Patient-derived induced pluripotent stem cells (iPSCs) were detected for human mitochondrial tRNAArg (hmtRNAArg) steady-state level, mitochondrial respiratory complex (MRC) activity, oxygen consumption rate (OCR), extracellular acidification rate (ECAR), mitochondrial membrane potential (MMP), reactive oxygen species (ROS) abundance, and apoptosis level. RESULTS: The three pedigrees were diagnosed as PCH6 caused by compound heterozygous RARS2 variations. Five RARS2 variants were identified: c.3G>C(p.M1?), c.685C>T(p.R229∗), c.1060T>A(p.F354I), c.1210A>G(p.M404V), and c.1369G>A(p.G457R). RARS2 c.3G>C disrupted protein expression. RARS2 c.685C>T created a truncated protein lacking complete catalytic core and anticodon-binding domain. RARS2 c.1060T>A and c.1369G>A were predicted to cause structural abnormality. The hmtRNAArg steady-state abundance in a patient's iPSCs was unaffected. Mitochondrial energy metabolism was normal, including MRC activity, OCR, ECAR, and MMP, while mitochondria-related cellular characteristics, including ROS (P < 0.001) and apoptosis levels (P < 0.001), increased. CONCLUSIONS: This study reports five RARS2 variations among which c.3G>C and c.1060T>A are novel. Summarized facial features of PCH6 patients will facilitate diagnosis. Defective mitochondrial energy metabolism may not be key points, but mitochondria-related abnormal cellular physiology, including apoptosis, may be an underlying pathogenesis.

A plasmon modulator by directly controlling the couple of photon and electron.

The manipulation of surface plasmon polaritons plays a pivotal role in plasmonic science and technology, however, the modulation efficiency of the traditional method suffers from the weak light-matter interaction. Herein, we propose a new method to overcome this obstacle by directly controlling the couple of photon and electron. In this paper, a hybrid graphene-dielectric- interdigital electrode structure is numerically and experimentally investigated. The plasmon is excited due to the confined carrier which is regulated by the potential wells. The frequency of plasmon can be tuned over a range of ~ 33 cm-1, and the obtained maximum extinction ratio is 8% via changing the confined area and the density of carrier. These findings may open up a new path to design the high efficiency all-optical modulator because the electrons can also be driven optically.

Impaired alpha rhythm in bad sleeper during nap: A high-density electroencephalogram study.

As the pace of life accelerates, poor sleep quality has become a common health problem. A wealth of sleep studies based on electroencephalogram (EEG) have found that poor sleep quality has decreased alpha rhythm during night-time sleep. However, whether the EEG rhythm during an afternoon nap has a similar impairment has seldom been investigated. By recruiting 35 subjects for a 90-min nap, we explored the relationship between sleep quality and the alpha rhythm, and the contribution of naps to subjective sleep experience. We found that the power of alpha rhythm in the occipital lobe negatively correlated with sleep quality. However, there was no correlation between the sleep quality at night and the sleep duration of the nap, nor the sleep quality and the proportion of each sleep period of a nap. Our present results indicated that the quality of night-time sleep did not affect the macrostructure of the nap. Interestingly, the negative correlation between the alpha rhythm of a nap and sleep quality at night continuously existed in Wake, N1 sleep, and N2 sleep, and had the most substantial value during N2 sleep of a nap. This implies that an impaired occipital lobe alpha rhythm is a robust biomarker for poor sleep. The research is of particular significance for developing electrical neural stimulation therapy to improve sleep quality, especially for the stimuli regions for selection and the time windows for implementation.

A Plant-Produced Recombinant Fusion Protein-Based Newcastle Disease Subunit Vaccine and Rapid Differential Diagnosis Platform.

Newcastle disease (ND) is a highly contagious avian disease, causing considerable economic losses to the poultry industry. To obtain a safe, inexpensive, and effective ND vaccine to meet the international trade requirements of differentiating infected from vaccinated animals (DIVA), here we report the production of Oryza sativa recombinant fusion (F) protein in stably transformed transgenic rice seeds via agroinfiltration. The F protein expression level was enhanced 3.6-fold with a genetic background in low glutelin. Inoculation of plant-produced F antigen into Specific Pathogen Free (SPF) chickens markedly elicited neutralizing antibody responses against homologous and heterologous ND virus strains. Two doses of 4.5 µg fully protected chickens from a lethal ND challenge without any clinical symptoms. The mean weight gain of F protein-immunized chickens within 15 days after challenge was significantly higher than that of traditional whole virus vaccine-immunized chickens, thereby obtaining higher economic benefits. Moreover, the sera from the chickens vaccinated with the plant-produced F vaccine did not show reactivity in an immunochromatographic strip targeting the haemagglutinin-neuraminidase protein (HN) protein, and DIVA could be achieved within 10 minutes. Our results demonstrate that the plant-derived F vaccine along with immunochromatographic strips could be useful in the implementation of an NDV eradication program.