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Abstract Introduction Wearing a helmet reduces the risk of head injuries substantially in the event of a motorcycle crash. Countries around the world are committed to promoting helmet use, but the progress has been slow and uneven. There is an urgent need for large-scale data collection for situation assessment and intervention evaluation. Methods This study proposes a scalable, low-cost algorithm to estimate helmet-wearing rates. Applying the state-of-the-art deep learning technique for object detection to images acquired from Google Street View, the algorithm has the potential to provide accurate estimates at the global level. Results Trained on a sample of 3995 images, the algorithm achieved high accuracy. The out-of-sample prediction results for all three object classes (helmets, drivers, and passengers) reveal a precision of 0.927, a recall value of 0.922, and a mean average precision at 50 (mAP50) of 0.956. Discussion The remarkable model performance suggests the algorithm’s capacity to generate accurate estimates of helmet-wearing rates from an image source with global coverage. The significant enhancement in the availability of helmet usage data resulting from this approach could bolster progress tracking and facilitate evidence-based policymaking for helmet wearing globally.

Abstract The field of regenerative medicine has witnessed remarkable advancements with the emergence of induced pluripotent stem cells (iPSCs) derived from a variety of sources. Among these, urine-derived induced pluripotent stem cells (u-iPSCs) have garnered substantial attention due to their non-invasive and patient-friendly acquisition method. This review manuscript delves into the potential and application of u-iPSCs in advancing precision medicine, particularly in the realms of drug testing, disease modeling, and cell therapy. U-iPSCs are generated through the reprogramming of somatic cells found in urine samples, offering a unique and renewable source of patient-specific pluripotent cells. Their utility in drug testing has revolutionized the pharmaceutical industry by providing personalized platforms for drug screening, toxicity assessment, and efficacy evaluation. The availability of u-iPSCs with diverse genetic backgrounds facilitates the development of tailored therapeutic approaches, minimizing adverse effects and optimizing treatment outcomes. Furthermore, u-iPSCs have demonstrated remarkable efficacy in disease modeling, allowing researchers to recapitulate patient-specific pathologies in vitro. This not only enhances our understanding of disease mechanisms but also serves as a valuable tool for drug discovery and development. In addition, u-iPSC-based disease models offer a platform for studying rare and genetically complex diseases, often underserved by traditional research methods. The versatility of u-iPSCs extends to cell therapy applications, where they hold immense promise for regenerative medicine. Their potential to differentiate into various cell types, including neurons, cardiomyocytes, and hepatocytes, enables the development of patient-specific cell replacement therapies. This personalized approach can revolutionize the treatment of degenerative diseases, organ failure, and tissue damage by minimizing immune rejection and optimizing therapeutic outcomes. However, several challenges and considerations, such as standardization of reprogramming protocols, genomic stability, and scalability, must be addressed to fully exploit u-iPSCs’ potential in precision medicine. In conclusion, this review underscores the transformative impact of u-iPSCs on advancing precision medicine and highlights the future prospects and challenges in harnessing this innovative technology for improved healthcare outcomes.

Adipose tissue (AT) serves as an energy-capacitive organ and performs functions involving paracrine- and endocrine-mediated regulation via extracellular vesicles (EVs) secretion. Exosomes, a subtype of EVs, contain various bioactive molecules with regulatory effects, such as nucleic acids, proteins, and lipids. AT-derived exosomes (AT-exos) include exosomes derived from various cells in AT, including adipocytes, adipose-derived stem cells (ADSCs), macrophages, and endothelial cells. This review aimed to comprehensively evaluate the impacts of different AT-exos on the regulation of physiological and pathological processes. The contents and functions of adipocyte-derived exosomes and ADSC-derived exosomes are compared simultaneously, highlighting their similarities and differences. The contents of AT-exos have been shown to exert complex regulatory effects on local inflammation, tumor dynamics, and insulin resistance. Significantly, differences in the cargoes of AT-exos have been observed among diabetes patients, obese individuals, and healthy individuals. These differences could be used to predict the development of diabetes mellitus and as therapeutic targets for improving insulin sensitivity and glucose tolerance. However, further research is needed to elucidate the underlying mechanisms and potential applications of AT-exos.

Abstract Background This study aimed to elucidate the methodology and assess the efficacy of the aortic arch inclusion technique using an artificial blood vessel in managing acute type A aortic dissection (ATAAD). Methods We conducted a retrospective review of 18 patients (11 males and 7 females, average age: 56.2 ± 8.6 years) diagnosed with ATAAD who underwent total aortic arch replacement (TAAR) using an artificial vascular “inclusion” between June 2020 and October 2022. During the operation, deep hypothermic circulatory arrest (DHCA) and selective antegrade cerebral perfusion (ACP) of the right axillary artery were employed for brain protection. The ‘inclusion’ total aortic arch replacement and stented elephant trunk (SET) surgery were performed. Results Four patients underwent the Bentall procedure during the study, with one additional patient requiring coronary artery bypass grafting (CABG) due to significant involvement of the right coronary orifice. Three patients died during postoperative hospitalization. Other notable complications included two cases of postoperative renal failure necessitating continuous renal replacement therapy (CRRT), one case of postoperative double lower limb paraplegia, and one case of cerebral infarction resulting in unilateral impairment of the left upper limb. Eleven patients underwent computed tomography angiography (CTA) examinations of the aorta three months to one-year post-operation. The CTA results revealed thrombosis in the false lumen surrounding the aortic arch stent in seven patients and complete thrombosis of the false lumen around the descending aortic stent in eight patients. One patient had partial thrombosis of the false lumen around the descending aortic stent, and another patient’s false lumen in the thoracic and abdominal aorta completely resolved after one year of follow-up. Conclusions Incorporating vascular graft in aortic arch replacement simplifies the procedure and yields promising short-term outcomes. It achieves the aim of total arch replacement using a four-branch prosthetic graft. However, extensive sampling and thorough, prolonged follow-up observations are essential to fully evaluate the long-term results.

Background: The stromal vascular fraction (SVF), a cluster of stem and progenitor cells isolated from adipose tissue, holds significant promise for application in regenerative medicine. However, the existing methods for SVF isolation are time-consuming and expensive. Thus, in this study, we explored a new method of SVF extraction—ultrasound-assisted SVF isolation (USASI)—and compared the viability and characteristics of SVF isolated using different methods. Methods: SVF extraction methods using different combinations of ultrasound power, ultrasound time, collagenase dosage, and collagenase digestion time were compared with those of the control group (collagenase digestion method). The cell yield and vitality of the SVF were evaluated via cell counting and trypan blue staining. The cell components and immunophenotypes of freshly isolated SVF were analyzed using flow cytometry. The proliferative capacity and differentiation potential of the SVF were also identified. Results: Ultrasonication at 95 ​W-20 kHz for 30 ​s followed by digestion with 0.15% collagenase for 30 ​min was identified as the most suitable parameter for the USASI method in isolating SVF, as recommended based on the evaluation of various tested conditions. The USASI method significantly reduced the collagenase dosage and shortened the digestion time. Compared to the collagenase digestion method, the USASI method had a higher cell yield and cell viability, with no adverse effects on cell components, proliferative capacity, or multipotential differentiation capacity. Conclusion: With reduced processing time, lower collagenase dosage, and increased cell yield without impairing the viability and characteristics of SVF, USASI holds the potential to emerge as a time-saving and cost-effective method for future clinical applications.

Abstract Skin fibrosis is characterized by the excessive accumulation of extracellular matrix (ECM) caused by fibrotic disorders of the skin. In recent years, ECM stiffness has emerged as a prominent mechanical cue that precedes skin fibrosis and drives its progression by promoting fibroblasts activation. However, how stiffness influences fibroblasts activation for skin fibrosis progression remains unknown. Here, we report a positive feedback loop mediated by the mechanosensitive ion channel Piezo1 and aberrant tissue mechanics in driving skin fibrosis. Piezo1 is upregulated in fibrotic skin in both humans and mice. Piezo1 knockdown dermal fibroblasts lose their fibroproliferative phenotypes despite being grown on a stiffer substrate. We show that Piezo1 acts through the Wnt2/Wnt11 pathway to mechanically induce secretion of C-C motif chemokine ligand 24 (CCL24, also known as eotaxin-2), a potent cytokine associated with fibrotic disorders. Importantly, adeno-associated virus (AAV)-mediated Piezo1 knockdown ameliorated the progression of skin fibrosis and skin stiffness in mice. Overall, increased matrix stiffness promotes skin fibrosis through the inflammatory Piezo1-Wnt2/Wnt11-CCL24 pathway. In turn, a stiffer skin microenvironment increases Piezo1 expression to exacerbate skin fibrosis aggression. Therefore, targeting Piezo1 represents a strategy to break the positive feedback loop between fibroblasts mechanotransduction and aberrant tissue mechanics in skin fibrosis.

Background: Melanocytic nevus is mainly treated by complete or partial removal. However, predicting the risk of malignant transformation of melanocytic nevi and which treatment patients should receive, surgical or non-surgical management, to gain the best results and aesthetic outcomes is controversial. Methods: Global literature on melanocytic nevus treatment, published between 1997 and 2022, was scanned using the Web of Science Core Collection database. Microsoft Office Excel, CiteSpace V, VOSviewer, Scimago Graphica, Bibliometrix, and Biblioshiny packages in R were used for the bibliometric analysis to summarize the leading countries, institutions, professors, and research trends in this field. Results: This study included 1 723 articles. Publications and citations exhibited positive trends over the past 20 years. The United States had the most productive organizations and publications in the comprehensive worldwide cooperation network, and China was recently one of the most active major participants. Professor Giovanni Pellacani, whose H-index, G-index, and M-index ranked first in this field, founded a virtual biopsy using reflectance confocal microscopy. In addition, Krengel and Kinsler contributed significantly to diagnosing and treating melanocytic nevi. The top 25 keywords in recent years were mostly about the mechanisms and risk factors for the malignant transformation of nevi. Conclusion: The future trend for melanocytic nevi treatment is to specify genotype-phenotype and genotype-outcome correlations, choose proper therapy to reduce the risk of malignant transformation, and simultaneously achieve the best aesthetic outcomes.

Abstract Background Bifid nose is a rare congenital deformity and the etiology is unknown. The purpose of this study was to report genetic variation in family of patients with bifid nose. Methods Twenty-three consecutive patients who were diagnosed with mild bifid nose were operated with z-plasty from 2009 to 2021. Three underage patients (a pair of twins and a girl) from two family lines, who came to our hospital for surgical treatment, were enrolled. Whole exome sequencing and Sanger sequencing were conducted. Z-shaped flaps were created and the cartilago alaris major were re-stitched. Photographs and CT scan before and after surgery were obtained. Clinical outcomes, complications and patients’ satisfaction were evaluated and analyzed. The follow-up time ranges from 2 to 3 years (2.4 ± 1.2 years). Results Most patients were satisfied with the outcome (96.2%). The nasal deformities were corrected successfully with z-plasty technique in one-stage. FREM1 c.870_876del and c.2 T > C were detected with Whole exome sequencing, which have not been reported before. The results of Sanger sequencing were consistent with those of Whole exome sequencing. Conclusions The newly detected mutations of FREM1 have a certain heritability, and are helpful to make an accurate diagnosis and provide a better understanding of bifid nose mechanism. Z-plasty technique can be an effective technical approach for correcting mild bifid nose deformity.

Abstract Background Wheat powdery mildew is an obligate biotrophic pathogen infecting wheat, which can pose a serious threat to wheat production. In this study, transcriptome sequencing was carried out on wheat leaves infected by Blumeria graminis f. sp. tritici from 0 h to 7 d. Results KEGG and GO enrichment analysis revealed that the upstream biosynthetic pathways and downstream signal transduction pathways of salicylic acid, jasmonic acid, and ethylene were highly enriched at all infection periods. Trend analysis showed that the expressions of hormone-related genes were significantly expressed from 1 to 4 d, suggesting that 1 d-4 d is the main period in which hormones play a defensive role. During this period of time, the salicylic acid pathway was up-regulated, while the jasmonic acid and ethylene pathways were suppressed. Meanwhile, four key modules and 11 hub genes were identified, most of which were hormone related. Conclusion This study improves the understanding of the dynamical responses of wheat to Blumeria graminis f. sp. tritici infestation at the transcriptional level and provides a reference for screening core genes regulated by hormones.

Tartary buckwheat (Fagopyrum tataricum) is a significant medicinal crop, with flavonoids serving as a crucial measure of its quality. Presently, the artificial cultivation of Tartary buckwheat yields low results, and the quality varies across different origins. Therefore, it is imperative to identify an effective method to enhance the yield and quality of buckwheat. Endophytic fungi reside within plants and form a mutually beneficial symbiotic relationship, aiding plants in nutrient absorption, promoting host growth, and improving secondary metabolites akin to the host. In this study, high-throughput sequencing technology was employed to assess the diversity of endophytic fungi in Tartary buckwheat. Subsequently, a correlation analysis was performed between fungi and metabolites, revealing potential increases in flavonoid content due to endophytic fungi such as Bipolaris, Hymenula, and Colletotrichum. Additionally, a correlation analysis between fungi and phenotypic traits unveiled the potential influence of endophytic fungi such as Bipolaris, Buckleyzyma, and Trichosporon on the phenotypic traits of Tartary buckwheat. Notably, the endophytic fungi of the Bipolaris genus exhibited the potential to elevate the content of Tartary buckwheat metabolites and enhance crop growth. Consequently, this study successfully identified the resources of endophytic fungi in Tartary buckwheat, explored potential functional endophytic fungi, and laid a scientific foundation for future implementation of biological fertilizers in improving the quality and growth of Tartary buckwheat.

Abstract Background N7-methylguanosine (m7G) is one of the most conserved modifications in nucleosides impacting mRNA export, splicing, and translation. However, the precise function and molecular mechanism of internal mRNA m7G methylation in adult hippocampal neurogenesis and neurogenesis-related Alzheimer’s disease (AD) remain unknown. Results We profiled the dynamic Mettl1/Wdr4 expressions and m7G modification during neuronal differentiation of neural stem cells (NSCs) in vitro and in vivo. Adult hippocampal neurogenesis and its molecular mechanisms were examined by morphology, biochemical methods and biological sequencing. The translation efficiency of mRNA was detected by polysome profiling. The stability of Sptbn2 mRNA was constructed by RNA stability assay. APPswe/PS1ΔE9 (APP/PS1) double transgenic mice were used as model of AD. Morris water maze was used to detect the cognitive function. Methods We found that m7G methyltransferase complex Mettl1/Wdr4 as well as m7G was significantly elevated in neurons. Functionally, silencing Mettl1 in neural stem cells (NSCs) markedly decreased m7G modification, neuronal genesis and proliferation in addition to increasing gliogenesis, while forced expression of Mettl1 facilitated neuronal differentiation and proliferation. Mechanistically, the m7G modification of Sptbn2 mRNA by Mettl1 enhanced its stability and translation, which promoted neurogenesis. Importantly, genetic defciency of Mettl1 reduced hippocampal neurogenesis and spatial memory in the adult mice. Furthermore, Mettl1 overexpression in the hippocampus of APP/PS1 mice rescued neurogenesis and behavioral defects. Conclusion Our findings unravel the pivotal role of internal mRNA m7G modification in Sptbn2-mediated neurogenesis, and highlight Mettl3 regulation of neurogenesis as a novel therapeutic target in AD treatment.

Background: Scar contractions caused by trauma or burns can cause secondary physical dysfunction and disfigurement. Many minimally invasive methods for scar contraction have shown limited applicability and efficacy. This study investigated the feasibility and efficacy of intralesional collagenase injections for scar contraction treatment. Methods: Patients with contracted scars who had limited joint movement and physical disfiguration for >1 year were enrolled in this single-blind, randomized clinical trial from July 2017 to February 2018 ​at Shanghai Ninth People’s Hospital. Collagenase was injected into the firm-contracted scar (15 U/cm2) three times at 4-week intervals in the multiple treatment group and once in the single treatment group, and a placebo injection was performed in the control group. Scar length and skin texture were documented at the 4- and 12-week follow-ups. The safety of the collagenase treatment was also evaluated. Results: The contracted scar was significantly elongated after both single and multiple collagenase treatments. The results showed that, compared to a one-time treatment, repeated injections were more effective at 12 weeks, with an average improvement of 26.83 (15.79%). At 12 weeks, 78.9% of the patients in the multiple group and 52.9% in the single group achieved significant improvement at 12 weeks. No severe adverse events were observed. Conclusion: Intralesional collagenase injection showed promising results in improving scar contraction and provides an alternative treatment for patients. Trial registration: ChiCTR, ChiCTR-IIR-16009910. Registered November 18, 2016. Prospective registration, https://www.chictr.org.cn/showproj.html?proj=16853.

Abstract The increased mechanics of fibrotic skin tissue continuously regulate fibroblast functions such as survival and differentiation. Although all these processes consume metabolites, it is unclear whether and how cells adapt their metabolic activity to increased matrix stiffness. Here, we show that transferring mouse dermal fibroblasts from soft to stiff substrates causes an up-regulation of arginine and proline metabolism. Increased matrix stiffness stimulates the expression and activity of key metabolic enzymes, leading to the synthesis of L-proline, a major source of collagen. In addition, the novel mechanosensitive channel Piezo1 was identified as a key regulator of arginine and proline metabolism in fibroblasts under increased stiffness. Consistently, targeting Piezo1 to dermal fibroblasts in vivo effectively reduces fibrosis and arginine-proline metabolism in mouse skin. Therefore, mechanical stiffness is a critical environmental cue for fibroblast metabolism and skin fibrosis progression.

Abstract Series compensation grids connected with type-3 wind turbine generator (WTG)-based wind farms have suffered numerous subsynchronous oscillation (SSO) events worldwide. For early alerting of SSO and effective development of protection and control strategies, it is critical to monitor and identify SSO accurately and quickly. Ambient data is continuously available, which is useful for online monitoring. This paper proposes an ambient data-driven SSO online monitoring method based on the Kalman filter (KF) combined with the multi-model partitioning filter (MMPF). The KF is utilized to fit the measured ambient data with an auto regressive (AR) model. Then, the damping factor (or damping ratio) and frequency in the SSO mode can be acquired by solving the roots of the characteristic polynomial corresponding to the AR model. Moreover, the MMPF is an effective model order selection method applied to the KF for better identification. The performance of the MMPF-KF method is demonstrated by simulations and real-time experiments. The results of case studies validate the effectiveness of the proposed method under various conditions.

Abstract Background Impaired wound re-epithelialization contributes to cutaneous barrier reconstruction dysfunction. Recently, N 6-methyladenosine (m6A) RNA modification has been shown to participate in the determination of RNA fate, and its aberration triggers the pathogenesis of numerous diseases. Howbeit, the function of m6A in wound re-epithelialization remains enigmatic. Methods Alkbh5 ‒/‒ mouse was constructed to study the rate of wound re-epithelialization after ALKBH5 ablation. Integrated high-throughput analysis combining methylated RNA immunoprecipitation sequencing (MeRIP-seq) and RNA-seq was used to identify the downstream target of ALKBH5. In vitro and in vivo rescue experiments were conducted to verify the role of the downstream target on the functional phenotype of ALKBH5-deficient cells or animals. Furthermore, the interacting reader protein and regulatory mechanisms were determined through RIP-qPCR, RNA pull–down, and RNA stability assays. Results ALKBH5 was specifically upregulated in the wound edge epidermis. Ablation of ALKBH5 suppressed keratinocyte migration and resulted in delayed wound re-epithelialization in Alkbh5 ‒/‒ mouse. Integrated high-throughput analysis revealed that PELI2, an E3 ubiquitin protein ligase, serves as the downstream target of ALKBH5. Concordantly, exogenous PELI2 supplementation partially rescued keratinocyte migration and accelerated re-epithelialization in ALKBH5-deficient cells, both in vitro and in vivo. In terms of its mechanism, ALKBH5 promoted PELI2 expression by removing the m6A modification from PELI2 mRNA and enhancing its stability in a YTHDF2-dependent manner. Conclusions This study identifies ALKBH5 as an endogenous accelerator of wound re-epithelialization, thereby benefiting the development of a reprogrammed m6A targeted therapy for refractory wounds. Graphical Abstract

Background:. Letibotulinum toxin A has an established efficacy and safety profile for aesthetic treatment of glabellar wrinkles. This study was conducted to demonstrate the noninferiority of letibotulinum toxin A versus onabotulinum toxin A in improving the appearance of moderate-to-severe glabellar wrinkles in Chinese patients. Methods:. This phase-III multicenter, randomized, parallel positive control, double-blinded study compared the efficacy and safety of letibotulinum toxin A and onabotulinum toxin A. Eligible participants were randomized 3:1 to receive 20 U of letibotulinum toxin A or onabotulinum toxin A and were observed for 16 weeks postinjection. The primary endpoint was noninferiority in the proportion of study participants receiving a score of 0 or 1 for glabellar wrinkles on a four-point photographic evaluation scale, as assessed by an institution evaluator at maximum frown at week 4. Secondary endpoints included assessments at rest, photographic assessment of efficacy, and subjective self-assessment of the study participants. Results:. The proportion of participants (N = 500) receiving a score of 0 or 1 at maximum frown by the institution evaluator at week 4 was 88.49% for letibotulinum toxin A and 87.39% for onabotulinum toxin A (difference, 1.10%; 95% confidence interval, −5.02 to 8.82; P = 0.7469). No significant differences were observed between the treatments for secondary efficacy or safety endpoints. Participants’ self-assessment and satisfaction tended to be higher for letibotulinum toxin A than onabotulinum toxin A. Conclusion:. Letibotulinum toxin A is noninferior to onabotulinum toxin A in improving the appearance of moderate-to-severe glabellar wrinkles in Chinese patients.

Background:. Soft-tissue fillers, specifically hyaluronic acid fillers, can reduce many signs of aging by treating the associated loss of subcutaneous fat and midfacial contour deficiencies. The objective of this study was to investigate whether the effectiveness and safety of Belotero Volume Lidocaine (BVL) compared with Restylane (RES, control) is noninferior in the treatment of severe nasolabial folds (NLFs) in Chinese patients. Methods:. This was a prospective, randomized, controlled, split-face clinical study. Overall, 220 Chinese patients of both sexes with symmetrical NLFs of severe intensity (grade 4) on the Wrinkle Severity Rating Scale (WSRS) were treated with both fillers. Treatment outcomes were assessed by the WSRS, and other scales, at multiple time points up to 18 months postinjection. The co-primary effectiveness outcomes were based on the blinded evaluator ratings of NLFs according to the WSRS scale after 6 and 12 months. Adverse events were assessed during the whole study and patients’ pain sensation at three time points after injection. Results:. Noninferiority of BVL versus control based on the WSRS was demonstrated at month 6 and month 12. Response rates were slightly higher for BVL than control at all time points, and BVL had a sustained effect until month 18. Pain sensation scores were significantly lower for BVL compared with control. The incidence rates of treatment-related AEs were low and very similar for both treatments. Conclusions:. This study demonstrates that BVL is a safe, long-lasting, and effective treatment to correct severe NLFs in Chinese patients while being noninferior to the control device.

Background: To investigate the common symptoms after Covid-19 infection, characteristics of adverse events after vaccination, changes in clinical manifestations related to Neurofibromatosis type 1 (NF1), as well as the current vaccination status and factors related to vaccine hesitation among NF1 patients, in order to provide a basis for scientific protection and vaccine acceptance in NF1 individuals in the new phase of pandemic management. Methods: From December 29, 2022, to January 10, 2023, we conducted a self-assessment questionnaire survey among diagnosed NF1 patients. General data were provided including sex, age, main clinical presentations, and current treatment. This study mainly focused on the infection and vaccination status of Covid-19 among these patients with NF1. The data were statistically analyzed using SPSS26.0 software. Results: Of the 250 questionnaires distributed, 226 were valid. Among the 164 patients (72.6%) with Covid-19 infection, the most common infection symptoms and incidence of patients were not significantly different from those in the normal population (P>0.05), but the incidence of symptoms such as nasal congestion, headache, myalgia, sore throat, abdominal pain, diarrhea, and eye discomfort was higher than that in the normal population (P0.05), but the incidence of fatigue and headache was higher in NF1 patients (P

Abstract Fibrosis, a process caused by excessive deposition of extracellular matrix (ECM), is a common cause and outcome of organ failure and even death. Researchers have made many efforts to understand the mechanism of fibrogenesis and to develop therapeutic strategies; yet, the outcome remains unsatisfactory. In recent years, advances in epigenetics, including chromatin remodeling, histone modification, DNA methylation, and noncoding RNA (ncRNA), have provided more insights into the fibrotic process and have suggested the possibility of novel therapy for organ fibrosis. In this review, we summarize the current research on the epigenetic mechanisms involved in organ fibrosis and their possible clinical applications. Graphical Abstract

Abstract Sphincter dysfunction often occurs at the end of tubule organs such as the urethra, anus, or gastroesophageal sphincters. It is the primary consequence of neuromuscular impairment caused by trauma, inflammation, and aging. Despite intensive efforts to recover sphincter function, pharmacological treatments have not achieved significant improvement. Cell- or growth factor-based therapy is a promising approach for neuromuscular regeneration and the recovery of sphincter function. However, a decrease in cell retention and viability, or the short half-life and rapid degradation of growth factors after implantation, remain obstacles to the translation of these therapies to the clinic. Natural biomaterials provide unique tools for controlled growth factor delivery, which leads to better outcomes for sphincter function recovery in vivo when stem cells and growth factors are co-administrated, in comparison to the delivery of single therapies. In this review, we discuss the role of stem cells combined with the controlled release of growth factors, the methods used for delivery, their potential therapeutic role in neuromuscular repair, and the outcomes of preclinical studies using combination therapy, with the hope of providing new therapeutic strategies to treat incontinence or sphincter dysfunction of the urethra, anus, or gastroesophageal tissues, respectively.

Abstract Tropomyosin receptor kinase B (TrkB), a transmembrane receptor protein, has been found to play a pivotal role in neural development. This protein is encoded by the neurotrophic receptor tyrosine kinase 2 (NTRK2) gene, and its abnormal activation caused by NTRK2 overexpression or fusion can contribute to tumour initiation, progression, and resistance to therapeutics in multiple types of neurogenic tumours. Targeted therapies for this mechanism have been designed and developed in preclinical and clinical studies, including selective TrkB inhibitors and pan‐TRK inhibitors. This review describes the gene structure, biological function, abnormal TrkB activation mechanism, and current‐related targeted therapies in neurogenic tumours.

This paper introduces a novel approach to bipedal robot gait generation by proposing a higher-order form through the parameter equation of first-order Bessel interpolation. The trajectory planning for the bipedal robot, specifically for stepping up or down stairs, is established based on a three-dimensional interpolation equation. The experimental prototype, Roban, is utilized for the study, and the structural sketch of a single leg is presented. The inverse kinematics expression for the leg is derived using kinematic methods. Employing a position control method, the angle information is transmitted to the robot’s joints, enabling the completion of both downstairs simulation experiments and physical experiments with the Roban prototype. The analysis of the experimental process reveals a noticeable phenomenon of hip and ankle joint tilting in the robot. This observation suggests that low-cost bipedal robots driven by servo motors exhibit low stiffness characteristics in their joints.

Abstract Skin fibrosis, a pathological process featured by fibroblast activation and extracellular matrix (ECM) deposition, makes a significant contribution to morbidity. Studies have identified biomechanics as the central element in the complex network of fibrogenesis that drives the profibrotic feedback loop. In this study, we found that the acetylation of α‐tubulin at lysine 40 (K40) was augmented in fibrotic skin tissues. Further analysis showed that α‐tubulin acetylation is required for fibroblast activation, including contraction, migration, and ECM deposition. More importantly, we revealed that biomechanics‐induced upregulation of K40 acetylation promotes fibrosis by mediating mechanosensitive Yes‐associated protein S127 dephosphorylation and its cytoplasm nucleus shuttle. Furthermore, we demonstrated that the knockdown of α‐tubulin acetyltransferase 1 could rescue the K40 acetylation upregulation caused by increased matrix rigidity and ameliorate skin fibrosis both in vivo and in vitro. Herein, we highlight the critical role of α‐tubulin acetylation in matrix stiffness‐induced skin fibrosis and clarify a possible molecular mechanism. Our research suggests α‐tubulin acetylation as a potential target for drug design and therapeutic intervention.

Abstract Aiming at the problem of stability control of surrounding rock in the process of upward mining of inclined coal seam gob-side entry retaining, the gob-side entry retaining of 5449 working face in Beipingdong coal mine is taken as the research background. Based on the study of theoretical model of gob-side entry retaining, the law of overburden movement and surrounding rock deformation in the process of gob-side entry retaining is obtained through similar simulation experiment and numerical simulation analysis, and the optimal optimization scheme of supporting structure is put forward. The main research results are as follows: in the process of coal seam mining, the center of gravity of the overlying strata on the whole working face will shift to the side of the goaf, and the supporting structure composed of roadside filling body and bolt effectively inhibits the deformation of the roadway, in which the roadside filling body plays a key role; the top bolt bears greater stress than the side bolt in the process of mining, so the top bolt mainly maintains the stability of the roadway, and the side bolt plays the role of strengthening the side and preventing the extrusion. The width of the roadside filling body should be about 2.5 m, and the plastic failure range can be well controlled under this width.

Abstract During wound healing, oxygen availability and the anti-inflammatory microenvironment play an important role in the formation of new tissue. However, providing continuous and controllable oxygen around the injured tissue while inhibiting inflammation and realizing the synergistic effect of oxygen supply and anti-inflammation is still a major problem affecting the regeneration and repair of wound tissue. Inspired by skin wound pathology and the inflammatory microenvironment, a photothermal response-assisted strategy was developed in this study. We prepared a composite hydrogel system of polydopamine-hyaluronic acid (PDA-HA) hydrogel-loaded calcium peroxide-indocyanine green combined with lauric acid and manganese dioxide (CaO2-ICG@LA@MnO2) nanoparticles that showed excellent photothermal performance under near-infrared (NIR) irradiation and realized the on-off release of oxygen and reactive oxygen species (ROS). Controllable and sustainable oxygen release can promote the regeneration and repair of damaged tissue, and the generated ROS can effectively inhibit the outbreak of inflammation at the initial stage of wound healing. We believe that the system we have developed can be used in a new approach for treating chronic wounds.

A review of the literature in commonly used retrieval systems generally yields minimal or no pertinent information on the early works of Chinese doctors. Herein, the early works in rhinoplasty by pioneers Dr. Maolian Hu (胡懋廉) and Dr. PC Nyi (倪葆春) were retrieved from the database of Chinese Medical Journal published before 1949 that overseas doctors could not gain access to online possibly due to the language barrier in collecting and processing those old professional data by main retrieval systyems. They published original articles on costal cartilage applications for correction of saddle noses in as early as 1939 and 1949, respectively, which represent the earliest attempts of treating nasal deformities with costal cartilages in China as well as in Asia. The private cosmetic surgical practices in rhinoplasty from the 1930s to the 1940s were recovered and briefly reviewed, and some of the most important clinical cases were presented. Other important events related to rhinoplasty, such as the establishment of the first national society of rhinoplasty in Chengdu, China in 2011 and the successful performance of the first national congress on rhinoplasty in Shanghai, China in 2012, were included.

Lowering the Pt loading while maintaining high fuel-cell performance is an important but difficult task. Here the authors report modified polymers with intrinsic microporosity as binder for preferential retention of phosphoric acid in the catalyst layer, which offers great opportunities to lower the catalyst loading for high-temperature proton exchange membrane fuel cell.

As an essential trace element, iron is a necessary micronutrient for most organisms. Conyza blinii (C. blinii) is a traditional Chinese medicine grown in Sichuan, China. We chose C. blinii as material to explore the way plants respond to iron. Our results showed that iron increased the content of endogenous auxin, glandular trichomes (GTs) density and the content of blinin, the characteristic diterpene in C. blinii. Most of the key enzymes in the blinin synthesis pathway (MEP) were upregulated under iron by RNA sequencing. CbHO-1 (Heme oxygenase gene) was upregulated under exogenous IAA treatment. Taken together, these results suggest that C. blinii respond to iron through auxin- terpenoids metabolism, and this process may be related to CbHO-1.

Highly pathogenic avian influenza viruses (HPAIV), such as H5N1, H5N6, and H7N9, have been reported to frequently infect humans, but acute encephalitis caused by HPAIV in humans has been rarely reported. We report the first critical case of acute encephalitis with mild pneumonia caused by the H5N6 virus. On January 25 of 2022, a 6-year-old girl with severe neurological symptoms was admitted to our hospital and rapidly developed into seizures and coma. Brain imaging showed abnormalities. Electroencephalogram (EEG) presented abnormal slow waves. Cerebrospinal fluid (CSF) contained elevated protein (1.64 g/L) and white cells (546 × 106/L). Laboratory investigations revealed abnormally elevated transaminases, lactate dehydrogenase, and cytokines in serum. A novel reassortant H5N6 virus was identified from the patient’s serum, CSF, and tracheal aspirate specimens. Phylogenic analysis indicated that this virus was a novel reassortant avian-origin influenza A (H5N6) virus that belonged to clade 2.3.4.4b. This patient was diagnosed with acute encephalitis and discharged from the hospital accompanied by a language barrier. An epidemiological investigation confirmed that wild waterfowls were the direct source of infection in this case. Our study highlights the urgent need to pay attention to acute encephalitis caused by HPAIV.

Abstract Repairing large segment bone defects is still a clinical challenge. Bone tissue prefabrication shows great translational potentials and has been gradually accepted clinically. Existing bone reconstruction strategies, including autologous periosteal graft, allogeneic periosteal transplantation, xenogeneic periosteal transplantation, and periosteal cell tissue engineering, are all clinically valuable treatments and have made significant progress in research. Herein, we reviewed the research progress of these techniques and briefly explained the relationship among in vivo microenvironment, mechanical force, and periosteum osteogenesis. Moreover, we also highlighted the importance of the critical role of periosteum in osteogenesis and explained current challenges and future perspective.

Abstract Reconstruction of posterior lamellar eyelids remains challenging due to their delicate structure, highly specialized function, and cosmetic concerns. Current clinically available techniques for posterior lamellar reconstruction mainly focus on reconstructing the contour of the eyelids. However, the posterior lamella not only provides structural support for the eyelid but also offers a smooth mucosal surface to facilitate globe movement and secrete lipids to maintain ocular surface homeostasis. Bioengineered posterior lamellar substitutes developed via acellular or cellular approaches have shown promise as alternatives to current therapies and encouraging outcomes in animal studies and clinical conditions. Here, we provide a brief reference on the current application of autografts, biomaterials, and tissue‐engineered substitutes for posterior lamellar eyelid reconstruction. We also shed light on future challenges and directions for eyelid regeneration strategies and offer perspectives on transitioning replacement strategies to regeneration strategies for eyelid reconstruction in the future.

Background: Preceding works reveal the function of long non-coding RNAs (abbreviated to lncRNAs) during non-small cell lung cancer (NSCLC) evolvement. We explored the profile and biological functions of the lncRNA LINC00638 in NSCLC. Methods: Reverse transcription-quantitative PCR examined LINC00638 level in NSCLC and corresponding non-tumor tissues, human normal lung epithelial cells BEAS-2B, and NSCLC cells (NCI–H460, HCC-827, A549, H1299, H1975, H460). The gain- and loss-of-function assay of LINC00638 ascertained its function in modulating the proliferation, apoptosis, and invasion of NSCLC cells (HCC-827 and H460). Bioinformatics analysis investigated the underlying mechanisms. Dual luciferase reporter gene and RNA immunoprecipitation (RIP) checked the interactions between LINC00638 and microRNA (miR)-541-3p, miR-541-3p and insulin receptor substrate 1 (IRS1). Results: LINC00638 was upregulated in NSCLC tissues by contrast to the profiles found in the corresponding non-tumor normal tissues, as well as in NSCLC cells vis-à-vis BEAS-2B cells. LINC00638 upregulation pertained to the poorer survival rates of NSCLC patients. Overexpressing LINC00638 augmented NSCLC cells’ proliferation, growth, migration, and invasion but inhibited their apoptosis, while down-regulating LINC00638 led to the opposite. miR-541-3p might be an underlying target of LINC00638, which targeted IRS1, inhibited NSCLC progression, and reversed the carcinogenic effects of LINC00638. Mechanistically, LINC00638/miR-541-3p regulated the IRS1/phosphoinositide 3-kinase (PI3K)/Akt signaling pathway. Repressing IRS1/2 using its inhibitor NT157 repressed LINC00638-mediated oncogenic effects. Conclusion: LINC00638 may function as an oncogene in NSCLC by modulating the miR-541-3p/IRS1/PI3K/Akt axis.

The dried tuber of Alisma orientale (Sam.) Juzep. (AOJ) is a traditional Chinese medicine with high medicinal value. The endophytic fungi of medicinal plants are a treasure house of natural compounds. However, there is a lack of research on the diversity and biological activity of endophytic fungi of AOJ. In this study, high-throughput sequencing technology was used to study the diversity of endophytic fungi in the roots and stems of AOJ, and endophytic fungi with a high output of phenols and flavonoids were screened by chromogenic reaction, and the antioxidant and antibacterial activities and chemical constituents of crude extracts of their fermentation broth were studied. A total of 3,426 amplicon sequence variants (ASVs) belonging to 9 phyla, 27 classes, 64 orders, 152 families, and 277 genera were identified from AOJ. There were significant differences in the endophytic fungal communities of AOJ roots and stems, as well as in the endophytic fungal communities of triangular AOJ and circular AOJ. In addition, 31 strains of endophytic fungi were isolated from AOJ, of which 6 strains had good antioxidant and antibacterial activities. The crude extract of YG-2 had the strongest free radical scavenging ability and bacteriostatic ability, and its IC50DPPH, IC50ABTS, and IC50⋅OH values were 0.009 ± 0.000 mg/mL, 0.023 ± 0.002 mg/mL, and 0.081 ± 0.006 mg/mL, respectively. The results of LC-MS showed that the main component of the crude extract of YG-2 was caffeic acid (10.12 μmol/g). Overall, the results of this study preliminarily elucidated the diversity and community composition of endophytic fungi of AOJ, indicating that AOJ endophytic fungi have abundant secondary metabolites and good antioxidant and antibacterial activities. This study provides an important reference for further research, development and utilization of AOJ endophytic fungi and a theoretical basis for the further development of the endophytic fungus YG-2 (Chaetomium globosum) as a source of antioxidants.

ObjectivesThis study aimed to explore the value of conventional ultrasound (CUS) and contrast-enhanced ultrasound (CEUS) radiomics to diagnose breast cancer and predict its molecular subtype.MethodA total of 170 lesions (121 malignant, 49 benign) were selected from March 2019 to January 2022. Malignant lesions were further divided into six categories of molecular subtype: (non-)Luminal A, (non-)Luminal B, (non-)human epidermal growth factor receptor 2 (HER2) overexpression, (non-)triple-negative breast cancer (TNBC), hormone receptor (HR) positivity/negativity, and HER2 positivity/negativity. Participants were examined using CUS and CEUS before surgery. Regions of interest images were manually segmented. The pyradiomics toolkit and the maximum relevance minimum redundancy algorithm were utilized to extract and select features, multivariate logistic regression models of CUS, CEUS, and CUS combined with CEUS radiomics were then constructed and evaluated by fivefold cross-validation.ResultsThe accuracy of the CUS combined with CEUS model was superior to CUS model (85.4% vs. 81.3%, p

While ultrafast laser welding is an appealing technique for bonding transparent workpieces, it is not applicable for joining silicon samples due to nonlinear propagation effects which dramatically diminish the possible energy deposition at the interface. It is demonstrated that these limitations can be circumvented by local absorption enhancement at the interface thanks to metallic nanolayer deposition. By combining the resulting exalted absorption with filament relocation during ultrafast laser irradiation, silicon samples can be efficiently joined. Shear joining strengths >4 MPa are obtained for 21 nm gold nanolayers without laser‐induced alteration of the transmittance. Such remarkable strength values hold promise for applications in microelectronics, optics, and astronomy.

Geological hazards, such as the frequent occurrence of rock bursts in deep mining, emphasize the critical necessity for the early warning and prediction of dynamic fractures in coal and rock masses, as well as the destabilization of the surrounding rock. This study delves into the mechanisms of electromagnetic radiation (EMR) signals and their synchronous coupling with micro-seismic (ME) signals. EMR and ME signals from rock specimens were systematically collected during the uniaxial compression fracture process using a dedicated monitoring and acquisition system. Employing the wavelet analysis method, the original data underwent reconstruction and denoising, while the EMR and ME spectra, derived through fast Fourier transform, were subjected to detailed scrutiny. The comprehensive analysis unveiled that EMR signals arising from rock fractures exhibited precise timing synchronization with ME signals. Moreover, the dominant frequencies of both signals are closely aligned within the low-frequency band, indicating a remarkable degree of similarity and homology. These findings establish an experimental basis for the development of monitoring and early warning systems geared toward assessing damage to coal and rock masses using EMR and ME signals.

The main objective of this study was to investigate the influence of Serendipita indica on the growth of Tartary buckwheat plants. This study highlighted that the roots of Tartary buckwheat can be colonized by S. indica and that this fungal endophyte improved plants height, fresh weight, dry weight, and grain yield. In the meantime, the colonization of S. indica in Tartary buckwheat leaves resulted in elevated levels of photosynthesis, plant hormone content, antioxidant enzyme activity, proline content, chlorophyll content, soluble sugars, and protein content. Additionally, the introduction of S. indica to Tartary buckwheat roots led to a substantial rise in the levels of flavonoids and phenols found in the leaves and seeds of Tartary buckwheat. In addition, S. indica colonization reduced the content of malondialdehyde and hydrogen peroxide when compared to non-colonized plants. Importantly, the drought tolerance of Tartary buckwheat plants is increased, which benefits from physiology and bio-chemical changes in plants after S. indica colonized. In conclusion, we have shown that S. indica can improve systematic resistance and promote the growth of Tartary buckwheat by enhancing the photosynthetic capacity of Tartary buckwheat, inducing the production of IAA, increasing the content of secondary metabolites such as total phenols and total flavonoids, and improving the antioxidant enzyme activity of the plant.

Background: Skin expansion is a useful method for harvesting extra tissue. However, the outcome is hardly predictable. Methods: A total of 158 patients with 277 expanded skin cases were reviewed and evaluated via photographs. The review and evaluation were conducted to determine the skin’s regenerative condition. The overall texture of the expanded skin, which was deemed good, fair, or poor, was evaluated. The occurrence of five indications of the limitation of skin regeneration (thinning, color change, stretch marks, varicose vessels, and skin lesions) during expansion was recorded. The correlation between the five indications and the overall texture was statistically analyzed. Results: Among the 277 retrospectively reviewed expansion cases, the occurrence rate of skin deterioration showed significant differences between the expansion sites (P

Abstract Hair conditions, such as hair loss and graying, are prevalent human conditions. But they are often poorly controlled due to our insufficient understanding of human scalp hair follicle (hsHF) in health and disease. Here we describe a comprehensive single-cell RNA-seq (scRNA-seq) analysis on highly purified black and early-stage graying hsHFs. Based on these, a concise single-cell atlas for hsHF and its early graying changes is generated and verified using samples from multiple independent individuals. These data reveal the lineage trajectory of hsHF in unprecedented detail and uncover its multiple unexpected features not found in mouse HFs, including the presence of an innerbulge like compartment in the growing phase, lack of a discrete companion layer, and enrichment of EMT features in HF stem cells (HFSCs). Moreover, we demonstrate that besides melanocyte depletion, early-stage human hair graying is also associated with specific depletion of matrix hair progenitors but not HFSCs. The hair progenitors’ depletion is accompanied by their P53 pathway activation whose pharmaceutical blockade can ameliorate hair graying in mice, enlightening a promising therapeutic avenue for this prevalent hair condition.

Skin fibrosis is a physiopathological process featuring the excessive deposition of extracellular matrix (ECM), which is the main architecture that provides structural support and constitutes the microenvironment for various cellular behaviors. Recently, increasing interest has been drawn to the relationship between the mechanical properties of the ECM and the initiation and modulation of skin fibrosis, with the engagement of a complex network of signaling pathways, the activation of mechanosensitive proteins, and changes in immunoregulation and metabolism. Simultaneous with the progression of skin fibrosis, the stiffness of ECM increases, which in turn perturbs mechanical and humoral homeostasis to drive cell fate toward an outcome that maintains and enhances the fibrosis process, thus forming a pro-fibrotic “positive feedback loop”. In this review, we highlighted the central role of the ECM and its dynamic changes at both the molecular and cellular levels in skin fibrosis. We paid special attention to signaling pathways regulated by mechanical cues in ECM remodeling. We also systematically summarized antifibrotic interventions targeting the ECM, hopefully enlightening new strategies for fibrotic diseases.

Within the ultrarelativistic quantum molecular dynamics (UrQMD) model, by reversely tracing nucleons that are finally emitted at mid-rapidity (|y0| < 0.1) in the entire reaction process, the time evolution of elliptic flow (v2) of these traced nucleons is studied in Au+Au collisions at beam energy of 0.4 GeV/nucleon with different impact parameters. The initial value of v2 is positive and increases with impact parameter, it then decreases as time passes and tends to saturate at a negative value. It is found that nucleon-nucleon collisions always suppress the value of v2 (enhance the out-of-plane emission), while the nuclear mean field potential effect is more complex, which depends on both impact parameter and reaction time. The most relevant density probed by v2 of nucleons at mid-rapidity is found to be ∼ 60% of the maximum density reached during the collisions.

BackgroundHighly effective COVID-19 vaccines are available and free of charge in the United States. With adequate coverage, their use may help return life back to normal and reduce COVID-19–related hospitalization and death. Many barriers to widespread inoculation have prevented herd immunity, including vaccine hesitancy, lack of vaccine knowledge, and misinformation. The Ad Council and COVID Collaborative have been conducting one of the largest nationwide targeted campaigns (“It’s Up to You”) to communicate vaccine information and encourage timely vaccination across the United States. More than 300 major brands, digital and print media companies, and community-based organizations support the campaigns to reach distinct audiences. ObjectiveThe goal of this study was to use aggregated mobility data to assess the effectiveness of the campaign on COVID-19 vaccine uptake. MethodsCampaign exposure data were collected from the Cuebiq advertising impact measurement platform consisting of about 17 million opted-in and deidentified mobile devices across the country. A Bayesian spatiotemporal hierarchical model was developed to assess campaign effectiveness through estimating the association between county-level campaign exposure and vaccination rates reported by the Centers for Disease Control and Prevention. To minimize potential bias in exposure to the campaign, the model included several control variables (eg, age, race or ethnicity, income, and political affiliation). We also incorporated conditional autoregressive residual models to account for apparent spatiotemporal autocorrelation. ResultsThe data set covers a panel of 3104 counties from 48 states and the District of Columbia during a period of 22 weeks (March 29 to August 29, 2021). Officially launched in February 2021, the campaign reached about 3% of the anonymous devices on the Cuebiq platform by the end of March, which was the start of the study period. That exposure rate gradually declined to slightly above 1% in August 2021, effectively ending the study period. Results from the Bayesian hierarchical model indicate a statistically significant positive association between campaign exposure and vaccine uptake at the county level. A campaign that reaches everyone would boost the vaccination rate by 2.2% (95% uncertainty interval: 2.0%-2.4%) on a weekly basis, compared to the baseline case of no campaign. ConclusionsThe “It’s Up to You” campaign is effective in promoting COVID-19 vaccine uptake, suggesting that a nationwide targeted mass media campaign with multisectoral collaborations could be an impactful health communication strategy to improve progress against this and future pandemics. Methodologically, the results also show that location intelligence and mobile phone–based monitoring platforms can be effective in measuring impact of large-scale digital campaigns in near real time.

IntroductionAnxiety disorder is the most common psychiatric disorder among adolescents, with generalized anxiety disorder (GAD) being a common subtype of anxiety disorder. Current studies have revealed abnormal amygdala function in patients with anxiety compared with healthy people. However, the diagnosis of anxiety disorder and its subtypes still lack specific features of amygdala from T1-weighted structural magnetic resonance (MR) imaging. The purpose of our study was to investigate the feasibility of using radiomics approach to distinguish anxiety disorder and its subtype from healthy controls on T1-weighted images of the amygdala, and provide a basis for the clinical diagnosis of anxiety disorder.MethodsT1-weighted MR images of 200 patients with anxiety disorder (including 103 GAD patients) as well as 138 healthy controls were obtained in the Healthy Brain Network (HBN) dataset. We extracted 107 radiomics features for the left and right amygdala, respectively, and then performed feature selection using the 10-fold LASSO regression algorithm. For the selected features, we performed group-wise comparisons, and use different machine learning algorithms, including linear kernel support vector machine (SVM), to achieve the classification between the patients and healthy controls.ResultsFor the classification task of anxiety patients vs. healthy controls, 2 and 4 radiomics features were selected from left and right amygdala, respectively, and the area under receiver operating characteristic curve (AUC) of linear kernel SVM in cross-validation experiments was 0.6739±0.0708 for the left amygdala features and 0.6403±0.0519 for the right amygdala features; for classification task for GAD patients vs. healthy controls, 7 and 3 features were selected from left and right amygdala, respectively, and the cross-validation AUCs were 0.6755±0.0615 for the left amygdala features and 0.6966±0.0854 for the right amygdala features. In both classification tasks, the selected amygdala radiomics features had higher discriminatory significance and effect sizes compared with the amygdala volume.DiscussionOur study suggest that radiomics features of bilateral amygdala potentially could serve as a basis for the clinical diagnosis of anxiety disorder.

ABSTRACT The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has been an unprecedented public health disaster in human history, and its spike (S) protein is the major target for vaccines and antiviral drug development. Although widespread vaccination has been well established, the viral gene is prone to rapid mutation, resulting in multiple global spread waves. Therefore, specific antivirals are needed urgently, especially those against variants. In this study, the domain of the receptor binding motif (RBM) and fusion peptide (FP) (amino acids [aa] 436 to 829; denoted RBMFP) of the SARS-CoV-2 S protein was expressed as a recombinant RBMFP protein in Escherichia coli and identified as being immunogenic and antigenically active. Then, the RBMFP proteins were used for phage display to screen the novel affibody. After prokaryotic expression and selection, four novel affibody molecules (Z14, Z149, Z171, and Z327) were obtained. Through surface plasmon resonance (SPR) and pseudovirus neutralization assay, we showed that affibody molecules specifically bind to the RBMFP protein with high affinity and neutralize against SARS-CoV-2 pseudovirus infection. Especially, Z14 and Z171 displayed strong neutralizing activities against Delta and Omicron variants. Molecular docking predicted that affibody molecule interaction sites with RBM overlapped with ACE2. Thus, the novel affibody molecules could be further developed as specific neutralization agents against SARS-CoV-2 variants. IMPORTANCE SARS-CoV-2 and its variants are threatening the whole world. Although a full dose of vaccine injection showed great preventive effects and monoclonal antibody reagents have also been used for a specific treatment, the global pandemic persists. So, developing new vaccines and specific agents are needed urgently. In this work, we expressed the recombinant RBMFP protein as an antigen, identified its antigenicity, and used it as an antigen for affibody phage-display selection. After the prokaryotic expression, the specific affibody molecules were obtained and tested for pseudovirus neutralization. Results showed that the serum antibody induced by RBMFP neutralized Omicron variants. The screened affibody molecules specifically bound the RBMFP of SARS-CoV-2 with high affinity and neutralized the Delta and Omicron pseudovirus in vitro. So, the RBMFP induced serum provides neutralizing effects against pseudovirus in vitro, and the affibodies have the potential to be developed into specific prophylactic agents for SARS-CoV-2 and its variants.

Bacterial growth, under laboratory conditions or in a natural environment, goes through different growth phases. Some gene expressions are regulated with respect to the growth phase, which allows bacteria to adapt to changing conditions. Among them, many gene transcriptions are controlled by RpoHI or RpoHII in Rhodobacter sphaeroides. In a previous study, it was proven that the alternative sigma factors, RpoE, RpoHI, and RpoHII, are the major regulators of oxidative stress. Moreover, the growth of bacteria reached a stationary phase, and following the outgrowth, rpoE, rpoHI, and rpoHII mRNAs increased with respect to the growth phase. In this study, we demonstrated the regulatory function of alternative sigma factors in the rsp_0557 gene. The gene rsp_0557 is expressed with respect to the growth phase and belongs to the RpoHI/RpoHII regulons. Reporter assays showed that the antisigma factor ChrR turns on or over the RpoE activity to regulate rsp_0557 expression across the growth phase. In the exponential phase, RpoHII and sRNA Pos19 regulate the expression of rsp_0557 to an appropriate level under RpoE control. In the stationary phase, RpoHI and Pos19 stabilize the transcription of rsp_0557 at a high level. During outgrowth, RpoHI negatively regulates the transcription of rsp_0557. Taken together, our data indicate that these regulators are recruited by cells to adapt to or survive under different conditions throughout the growth phase. However, they still did not display all of the regulators involved in growth phase-dependent regulation. More research is still needed to learn more about the interaction between the regulators and the process of adapting to changed growth conditions and environments.