Motor neurons generate pose-targeted movements via proprioceptive sculpting.

Motor neurons are the final common pathway1 through which the brain controls movement of the body, forming the basic elements from which all movement is composed. Yet how a single motor neuron contributes to control during natural movement remains unclear. Here we anatomically and functionally characterize the individual roles of the motor neurons that control head movement in the fly, Drosophila melanogaster. Counterintuitively, we find that activity in a single motor neuron rotates the head in different directions, depending on the starting posture of the head, such that the head converges towards a pose determined by the identity of the stimulated motor neuron. A feedback model predicts that this convergent behaviour results from motor neuron drive interacting with proprioceptive feedback. We identify and genetically2 suppress a single class of proprioceptive neuron3 that changes the motor neuron-induced convergence as predicted by the feedback model. These data suggest a framework for how the brain controls movements: instead of directly generating movement in a given direction by activating a fixed set of motor neurons, the brain controls movements by adding bias to a continuing proprioceptive-motor loop.

Reliability and validity of the Chinese revised version of the pectus excavatum evaluation questionnaire in children with pectus excavatum.

OBJECTIVE: This study aims to translate the Pectus Excavatum Evaluation Questionnaire(PEEQ) into Chinese, and to comprehensively assess subjective outcomes in quality of life of children with pectus excavatum. METHODS: The PEEQ was translated from English to Chinese as according to the PRO translation guidelines. Structural validity and reliability of the questionnaire were examined by validated factor analysis and Cronbach's alpha coefficient analysis respectively. RESULTS: The results of the validation factor analysis for the Chinese PEEQ parent's and child's questionnaires demonstrated that the fit indicators for each dimension met the required criteria. The overall Cronbach's alpha coefficient of parent's and child's questionnaires were 0.840 and 0.854. Both the item-level content validity index (I-CVI) and scale-level content validity index (S-CVI) of each sub-questionnaire were 1. CONCLUSION: The Chinese version of the PEEQ parent's questionnaire is suitable as a proxy assessment for patients with PE, but the child's questionnaire needs further adjustments.

Elevated exosome-transferrable lncRNA EPB41L4A-AS1 in CD56bright NK cells is responsible for the impaired NK function in neuroblastoma patients by suppressing cell glycolysis.

NK cells are one of key immune components in neuroblastoma (NB) surveillance and eradication. Glucose metabolism as a major source of fuel for NK activation is exquisitely regulated. Our data revealed a diminished NK activation and a disproportionally augmented CD56bright subset in NB. Further study showed that NK cells in NB presented with an arrested glycolysis accompanied by an elevated expression of the long noncoding RNA (lncRNA) EPB41L4A-AS1, a known crucial participant in glycolysis regulation, in the CD56bright NK subset. The inhibitory function of lncRNA EPB41L4A-AS1 was recapitulated. Interestingly, our study demonstrated that exosomal lncRNA EPB41L4A-AS1 was transferrable from CD56bright NK to CD56dim NK and was able to quench the glycolysis of target NK. Our data demonstrated that an arrested glycolysis in patient NK cells was associated with an elevated lncRNA in CD56bright NK subset and a cross-talk between heterogeneous NK subsets was achieved by transferring metabolic inhibitory lncRNA through exosomes.

Plate Load Tests on an Unsaturated Sand-Kaolin Mixture with Varying Water Table.

Clayey sand is widely distributed and commonly encountered in geotechnical engineering practice. To understand its bearing capacity behavior under unsaturated conditions, plate load tests are performed on sand-kaolin mixture samples with varying water tables. The distributions of suction and volumetric water content with depth are measured by vibrating wire piezometers and soil moisture sensors, respectively. It is shown by the test results that the bearing capacity increases when the water table in the soil sample drops. The influence of suction on the bearing capacity is found to be dependent on the height of the water table and the hydraulic loading history of the soil sample. The plate load test results are interpreted using bearing capacity equations. Good agreement is obtained between measured and calculated bearing capacities. This study provides a simple method to estimate the bearing capacity of in situ unsaturated soil foundations.

Drosophila Ionotropic Receptor 25a mediates circadian clock resetting by temperature.

Circadian clocks are endogenous timers adjusting behaviour and physiology with the solar day. Synchronized circadian clocks improve fitness and are crucial for our physical and mental well-being. Visual and non-visual photoreceptors are responsible for synchronizing circadian clocks to light, but clock-resetting is also achieved by alternating day and night temperatures with only 2-4 °C difference. This temperature sensitivity is remarkable considering that the circadian clock period (~24 h) is largely independent of surrounding ambient temperatures. Here we show that Drosophila Ionotropic Receptor 25a (IR25a) is required for behavioural synchronization to low-amplitude temperature cycles. This channel is expressed in sensory neurons of internal stretch receptors previously implicated in temperature synchronization of the circadian clock. IR25a is required for temperature-synchronized clock protein oscillations in subsets of central clock neurons. Extracellular leg nerve recordings reveal temperature- and IR25a-dependent sensory responses, and IR25a misexpression confers temperature-dependent firing of heterologous neurons. We propose that IR25a is part of an input pathway to the circadian clock that detects small temperature differences. This pathway operates in the absence of known 'hot' and 'cold' sensors in the Drosophila antenna, revealing the existence of novel periphery-to-brain temperature signalling channels.

An Improved Large-Scale Stress-Controlled Apparatus for Long-Term Seepage Study of Coarse-Grained Cohesive Soils.

Clay-gravel mixture has been widely used in high embankment dams and understanding its seepage characteristics is critical to dam safety. From the instrumental perspective, the realization of continuous pressurized water supply becomes a key technical challenge, significantly restricting the working conditions replicated in previous seepage apparatuses. To this end, a novel water provision system, relying on parallel-disposed sensor-based pressure devices, was introduced, so that the application of an existing large-scale stress-controlled apparatus can be expanded to long-term seepage tests regarding coarse-grained cohesive soils. Constant-head permeability tests were conducted on original-graded clay-gravel mixtures to investigate their hydraulic properties, incorporating the influence of stress relaxation. Test results show that with 35% gravel content, the clay-gravel mixture is suitable for dam construction as the core material. The stress relaxation holds a marginal effect on the hydraulic conductivity of soil. The functionality of this improved apparatus is verified, especially under long-term seepage conditions.

Gradient Waveguide Thickness Guided-Mode Resonance Biosensor.

Portable systems for detecting biomolecules have attracted considerable attention, owing to the demand for point-of-care testing applications. This has led to the development of lab-on-a-chip (LOC) devices. However, most LOCs are developed with a focus on automation and preprocessing of samples; fluorescence measurement, which requires additional off-chip detection instruments, remains the main detection method in conventional assays. By incorporating optical biosensors into LOCs, the biosensing system can be simplified and miniaturized. However, many optical sensors require an additional coupling device, such as a grating or prism, which complicates the optical path design of the system. In this study, we propose a new type of biosensor based on gradient waveguide thickness guided-mode resonance (GWT-GMR), which allows for the conversion of spectral information into spatial information such that the output signal can be recorded on a charge-coupled device for further analysis without any additional dispersive elements. A two-channel microfluidic chip with embedded GWT-GMRs was developed to detect two model assays in a buffer solution: albumin and creatinine. The results indicated that the limit of detection for albumin was 2.92 µg/mL for the concentration range of 0.8-500 µg/mL investigated in this study, and that for creatinine it was 12.05 µg/mL for the concentration range of 1-10,000 µg/mL. These results indicated that the proposed GWT-GMR sensor is suitable for use in clinical applications. Owing to its simple readout and optical path design, the GWT-GMR is considered ideal for integration with smartphones or as miniaturized displays in handheld devices, which could prove beneficial for future point-of-care applications.

In vivo bioassay to test the pathogenicity of missense human AIP variants.

BACKGROUND: Heterozygous germline loss-of-function mutations in the aryl hydrocarbon receptor-interacting protein gene (AIP) predispose to childhood-onset pituitary tumours. The pathogenicity of missense variants may pose difficulties for genetic counselling and family follow-up. OBJECTIVE: To develop an in vivo system to test the pathogenicity of human AIP mutations using the fruit fly Drosophila melanogaster. METHODS: We generated a null mutant of the Drosophila AIP orthologue, CG1847, a gene located on the Xchromosome, which displayed lethality at larval stage in hemizygous knockout male mutants (CG1847exon1_3 ). We tested human missense variants of 'unknown significance', with 'pathogenic' variants as positive control. RESULTS: We found that human AIP can functionally substitute for CG1847, as heterologous overexpression of human AIP rescued male CG1847exon1_3 lethality, while a truncated version of AIP did not restore viability. Flies harbouring patient-specific missense AIP variants (p.C238Y, p.I13N, p.W73R and p.G272D) failed to rescue CG1847exon1_3 mutants, while seven variants (p.R16H, p.Q164R, p.E293V, p.A299V, p.R304Q, p.R314W and p.R325Q) showed rescue, supporting a non-pathogenic role for these latter variants corresponding to prevalence and clinical data. CONCLUSION: Our in vivo model represents a valuable tool to characterise putative disease-causing human AIP variants and assist the genetic counselling and management of families carrying AIP variants.

Improving image quality with model-based iterative reconstruction algorithm for chest CT in children with reduced contrast concentration.

OBJECTIVE: To evaluate model-based iterative reconstruction (MBIR) in improving the image quality of chest CT in children with reduced concentration contrast medium (CM). METHODS: Fifty-six children (median age of 4 years) who received low-dose enhanced chest CT were enrolled as the study group and compared with the control group of 56 children. Both groups used the automatic tube current modulation to achieve age-based noise index values of 11-15 HU. The study group used 100 kVp and reduced CM concentration of 270 mgI/ml, and the images in this group were reconstructed with 50% adaptive statistical iterative reconstruction (ASIR) and MBIR. The control group used 120 kV and standard CM of 320 mgI/ml, and the images in this group were reconstructed with ASIR only. Subjective image quality and objective image quality of the three image sets were evaluated. The subjective quality included overall image noise, enhancement degree, lesion (including mediastinum mass, pulmonary space-occupying lesions, and parenchymal infiltrative lesions) conspicuity, and beam-hardening artifacts. The objective quality included the measurement of noise in the left ventricle and back muscle to calculate signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) of ventricle. RESULTS: There was no difference in radiation dose between the study (CTDIvol of 1.79 ± 1.45 mGy) and control (1.68 ± 0.92 mGy) groups (p = 0.65). However, the study group used 19.7% lower CM dose than the control group (5.84 ± 2.69 vs. 7.27 ± 3.80 gI), and the enhancement in all images met the diagnostic requirements. MBIR reduced image noise by 58.6% and increased SNR and CNR by 143.6% and 165.7%, respectively, compared to ASIR images in the control group. The two ASIR image sets had similar image quality. CONCLUSION: MBIR improved the image quality of low-radiation-dose chest CT in children at 19.3% reduced CM dose.

Mediastinal teratoma presenting with hemoptysis and pleuritis misdiagnosed as tuberculosis (empyema).

BACKGROUND: Mediastinal teratoma is uncommon in children. It can be very difficult to diagnose especially in early stage. Rarely, teratoma may rupture into adjacent structures and lead to lung lesions or pleuritis. The main rarity of our reported cases was the dynamic imaging findings very similar to the developmental process of tuberculosis in patients 1 and 2, the pachypleuritis in patients 2 and 3, the extremely elevated inflammatory markers very similar to empyema in patient 3, and the extremely atypical tumor shape in all patients. CASE PRESENTATION: We present three pediatric patients presenting predominantly with recurrent hemoptysis and/or chest pain who were ultimately diagnosed with mediastinal teratoma containing pancreatic tissue. All three patients were initially suspected to have tuberculosis or empyema, and underwent relevant treatment, but without improvement. Patient 1 had left hilar enlargement, and subsequently an enlarging calcified cavity within high-density consolidation was identified. Patient 2 initially presented with right-sided pulmonary consolidation and pleuritis, and subsequently developed right lower lobe calcification, pleural thickening, and irregular soft tissue in the right inferior mediastinum. Patient 3 was initially found to have right lobe consolidation accompanied by a massive right-sided pleural effusion with extremely elevated inflammatory markers in serum and pleural effusion. The effusion later acquired heterogeneous density and appeared to become encapsulated. In patients 2 and 3, pleural biopsy identified fibrous tissue (with and without granuloma). Thoracotomy/thoracoscopy revealed mediastinal teratoma in each case, all of which were completely excised and the patients made uneventful recoveries. Histopathologic analysis revealed mature cystic-solid teratoma containing pancreatic tissue in all patients, and calcification in patients 1 and 2. CONCLUSIONS: Clinicians should be mindful that mediastinal teratoma is a potential cause of hemoptysis, lung lesions and pleuritis. Calcification and pachypleuritis on chest imaging especially in patients without fever should be highly suspected of mediastinal teratoma. Pleural biopsy sometimes fails to assist in making a definitive diagnosis.

[Automatic diagnosis of pectus carinatum for children based on the improved Haller index].

Pectus carinatum (PC) is one of the most common chest wall anomalies, which is characterized by the protrusion of the anterior chest wall including the sternum and adjacent costal cartilages. Mildly patients suffer from mental problems such as self-abasement, while severely suffering patients are disturbed by significant cardiopulmonary symptoms. The traditional Haller index, which is widely used clinically to evaluate the severity of PC, is deficient in diagnosis efficiency and classification. This paper presents an improved Haller index algorithm for PC: first, the contour of the patient chest in the axial computed tomography (CT) slice where the most convex thorax presents is extracted; and then a cubic B-spline curve is employed to fit the extracted contour followed by an eclipse fitting procedure; finally, the improved Haller index and the classification index are automatically calculated based on the analytic curves. The results of CT data analysis using 22 preoperative and postoperative patient CT datasets show that the proposed diagnostic index for PC can diagnose and classify PC patients correctly, which confirms the feasibility of the evaluation index. Furthermore, digital measurement techniques can be employed to improve the diagnostic efficiency of PC, achieving one small step towards the computer-aided intelligent diagnosis and treatment for pediatric chest wall malformations.

Pyruvatibacter mobilis gen. nov., sp. nov., a marine bacterium from the culture broth of Picochlorum sp. 122.

A Gram-stain-negative, aerobic bacterium, designated strain GYP-11T, was isolated from the culture broth of a marine microalga, Picochloruma sp. 122. Cells were dimorphic rods; free living cells were motile by means of a single polar flagellum, and star-shaped-aggregate-forming cells were attached with stalks and non-motile. Sodium pyruvate or Tween 20 was required for growth on marine agar 2216.16S rRNA gene sequence analysis revealed that this isolate shared 94.07 % similarity with its closest type strain, Parvibaculum hydrocarboniclasticum EPR92T. Phylogenetic analyses indicated that strain GYP-11T represents a distinct lineage in a robust clade consisting of strain GYP-11T, alphaproteobacterium GMD21A06 and Candidatus Phaeomarinobacter ectocarpi Ec32. This clade was close to the genera Parvibaculum and Tepidicaulis in the order Rhizobiales. Chemotaxonomic and physiological characteristics, including cellular fatty acids and carbon source profiles, also readily distinguished strain GYP-11T from all established genera and species. Thus, it is concluded that strain GYP-11T represents a novel species of a new genus in the order Rhizobiales, for which the name Pyruvatibacter mobilis gen. nov., sp. nov. is proposed. The type strain of Pyruvatibacter mobilis is GYP-11T ( = CGMCC 1.15125T = KCTC 42509T).

Gelatiniphilus marinus gen. nov., sp. nov., a bacterium from the culture broth of a microalga, Picochlorum sp. 122, and emended description of the genus Hwangdonia.

A Gram-stain-negative, non-motile, non-spore-forming, rod-shaped bacterium, designated strain GYP-24T, was isolated from the culture broth of a marine microalga, Picochlorum sp. 122. Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain GYP-24T forms a robust cluster with H.wangdoniaseohaensis KCTC 32177T (95.8 % sequence similarity) in the family Flavobacteriaceae. Growth of strain GYP-24T was observed at 15, 22, 28, 30, 33 and 37 °C (optimal 30-33 °C), pH 6.0-10.0 (optimal pH 7.0-8.0) and in the presence of 0.5-4 % (w/v) NaCl (optimal 2-3 %). The only menaquinone of strain GYP-24T was MK-6, and the G+C content of the genomic DNA was 36.9 mol%. The major fatty acid profile comprised iso-C17 : 0 3-OH, summed feature 3 (C16 : 1 ω7c/ω6c), iso-C15 : 1 G and iso-C15 : 0. The major polar lipids of strain GYP-24T were phosphatidylethanolamine, one unidentified phospholipid, three unidentified aminolipids and three unidentified lipids. Comprehensive analyses based on polyphasic characterization of GYP-24T indicated that it represents a novel species of a new genus, for which the name Gelatiniphilus marinus gen. nov., sp. nov. is proposed. The type strain is GYP-24T (=KCTC 42903T=MCCC 1K01730T). An emended description of the genus Hwangdonia is also given.

Maribacter flavus sp. nov., isolated from a cyanobacterial culture pond.

A Gram-stain-negative, yellow, non-spore-forming, strictly aerobic bacterium, designated C3T, was isolated from a cyanobacterial culture pond. Cells were halophilic, rod-shaped and able to move by gliding. Growth of strain C3T was observed at 15-30 °C (optimum 25 °C), pH 6.0-9.0 (optimum pH 7.5), and in the presence of 1-7 % (w/v) NaCl (optimum 2-3 %). Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain C3T formed a distinct lineage within the family Flavobacteriaceae and exhibited the highest similarity (95.21 %) to the type strains of Maribacter dokdonensis, Maribacter arcticus, Maribacter orientalis and Maribacter stanieri, and 'Maribacter caenipelagi' HD-44. The only isoprenoid quinone present within strain C3T was menaquinone 6 (MK-6). The G+C content of genomic DNA was 41.5 mol%. The major polar lipids were phosphatidylethanolamine and three unidentified lipids. The predominant cellular fatty acids (>5 % of the total fatty acids) were iso-C15 : 1 G, iso-C15 : 0, iso-C15 : 0 3-OH, iso-C17 : 0 3-OH, and summed feature 3 (comprising C16 : 1ω7c and/or C16 : 1ω6c). On the basis of the phenotypic, chemotaxonomic and phylogenetic characteristics, strain C3T represents a novel species of the genus Maribacter, for which the name Maribacter flavus sp. nov. is proposed. The type strain is C3T ( = KCTC 42508T = CGMCC 1.15112T).

Aliikangiella marina gen. nov., sp. nov., a marine bacterium from the culture broth of Picochlorum sp. 122, and proposal of Kangiellaceae fam. nov. in the order Oceanospirillales.

A Gram-stain-negative, non-motile, non-spore-forming, long rod-shaped bacterium, designated strain GYP-15T, was isolated from the culture broth of a marine microalga, Picochloruma sp. 122. Phylogenetic analyses revealed that strain GYP-15T shared 90.6 % 16S rRNA gene sequence similarity with its closest relative, Kangiella aquimarina KCTC 12183T, and represents a distinct phylogenetic lineage in a robust clade consisting of GYP-15T and members of the genera Kangiella and Pleionea in the order Oceanospirillales. Chemotaxonomic and physiological characteristics, including major cellular fatty acids, NaCl tolerance and pattern of carbon source utilization, could also readily distinguish strain GYP-15T from all established genera and species. Thus, it is concluded that strain GYP-15T represents a novel species of a new genus, for which the name Aliikangiella marina gen. nov., sp. nov. is proposed. The type strain of Aliikangiella marina is GYP-15T ( = MCCC 1K01163T = KCTC 42667T). Based on phylogenetic results, 16S rRNA gene signature nucleotide pattern and some physiological characteristics, the three genera Kangiella, Pleionea and Aliikangiella are proposed to make up a novel family, Kangiellaceae fam. nov., in the order Oceanospirillales.

Wenzhouxiangella marina gen. nov, sp. nov, a marine bacterium from the culture broth of Picochlorum sp. 122, and proposal of Wenzhouxiangellaceae fam. nov. in the order Chromatiales.

A Gram-stain negative, non-motile, non-phototrophic, non-alkaliphilic, obligately aerobic, chemoheterotrophic, and rod-shaped bacterium, designated strain Ma-11(T), was isolated from the culture broth of a marine microalga, Picochloruma sp. 122. Phylogenetic analyses showed that strain Ma-11(T) has less than 91 % similarity to its closest relative, Thioalkalivibrio sulfidiphilus HL-EbGR7(T), represents a distinct phylogenetic lineage in the order Chromatiales, and could not be assigned to any defined families in this order. Chemotaxonomic, genetic and physiological characteristics, including major fatty acids, genomic G+C content, lack of motility, aerophilicity and chemoheterotrophicity, could readily distinguish strain Ma-11(T) from any established members of the order Chromatiales. Based on the 16S rRNA gene sequence analysis and its signature nucleotide pattern, a new family Wenzhouxiangellaceae fam. nov. comprising the genus Wenzhouxiangella gen. nov. and species Wenzhouxiangella marina sp. nov. is proposed. The type strain is Ma-11(T) (=CGMCC 1.14936(T) = KCTC 42284(T) = MCCC 1K00261(T)).

Experimental study on soil improvement by electrochemical injection of calcium chloride solutions with time interval.

An improved electroosmotic method is proposed in this paper to enhance the non-uniform effect and efficiency of electroosmotic process. Such method is electroosmotic flow with injection of calcium chloride through the anode, followed by injection through the central tube (a tube at the midpoint between the anode and the cathode) with a suitable time interval between injections. Experimental results indicate that using this method can significantly improve the non-uniform reduction in water content throughout the soil, mitigate the formation of cracks in the anode section, and therefore considerably inhibit the increase in the electric resistance. After treatment, the drained water could be raised to 3.59 times more than that of pure electroosmotic flow, and 1.3 times that of simultaneous injection through both the anode and the central tube with considerably slight increase in power consumption. Moreover, the area of cementation was also expanded, approximately twice larger than that of pure electroosmotic flow and one and a half that of simultaneous injection. It is also worth noting that the proposed method performs better with the same power consumption. The results demonstrate that electroosmotic flow with a suitable time interval between injections could improve the efficiency of electroosmotic process and expand the treatment region in soils, hence can be a promising and economic technique for soil improvement in practical engineering.

Global progress in clinical research on human respiratory syncytial virus vaccines.

Human respiratory syncytial virus (hRSV) not only affects newborns but also older adults, contributing to a substantial worldwide burden of disease. However, only three approved hRSV vaccines remain commercially available to date. The development of a safe, practical and broad-spectrum vaccine suitable for all age groups remains extremely challenging. Using five different approaches-live-attenuated, recombinant-vector, subunit, particle-based, and mRNA-nearly 30 hRSV vaccine candidates are currently conducting clinical trials worldwide; moreover, > 30 vaccines are under preclinical evaluation. This review presents a comprehensive overview of these hRSV vaccines along with prospects for the development of infectious disease vaccines in the post-COVID-19 pandemic era.

Functional Analyses of Four Cryptochromes From Aquatic Organisms After Heterologous Expression in Drosophila melanogaster Circadian Clock Cells.

Cryptochromes (Crys) represent a multi-facetted class of proteins closely associated with circadian clocks. They have been shown to function as photoreceptors but also to fulfill light-independent roles as transcriptional repressors within the negative feedback loop of the circadian clock. In addition, there is evidence for Crys being involved in light-dependent magneto-sensing, and regulation of neuronal activity in insects, adding to the functional diversity of this cryptic protein class. In mammals, Crys are essential components of the circadian clock, but their role in other vertebrates is less clear. In invertebrates, Crys can function as circadian photoreceptors, or as components of the circadian clock, while in some species, both light-receptive and clock factor roles coexist. In the current study, we investigate the function of Cry proteins in zebrafish (Danio rerio), a freshwater teleost expressing 6 cry genes. Zebrafish peripheral circadian clocks are intrinsically light-sensitive, suggesting the involvement of Cry in light-resetting. Echinoderms (Strongylocentrotus purpuratus) represent the only class of deuterostomes that possess an orthologue (SpuCry) of the light-sensitive Drosophila melanogaster Cry, which is an important component of the light-resetting pathway, but also works as transcriptional repressor in peripheral clocks of fruit flies. We therefore investigated the potential of different zebrafish cry genes and SpuCry to replace the light-resetting and repressor functions of Drosophila Cry by expressing them in fruit flies lacking endogenous cry function. Using various behavioral and molecular approaches, we show that most Cry proteins analyzed are able to fulfill circadian repressor functions in flies, except for one of the zebrafish Crys, encoded by cry4a. Cry4a also shows a tendency to support light-dependent Cry functions, indicating that it might act in the light-input pathway of zebrafish.

Therapeutic SHPRH-146aa encoded by circ-SHPRH dynamically upregulates P21 to inhibit CDKs in neuroblastoma.

Recent research has underscored the significance of circular RNAs (circRNAs) in various cancers, including neuroblastoma (NB). Specifically, circ-SHPRH, a unique circRNA, has been revealed to inhibit tumor growth by sequestering miRNAs or producing the SHPRH-146aa protein. To explore circ-SHPRH's involvement in NB and its potential application in gene therapy, this study examined circ-SHPRH expression in 94 NB tissues and cell lines (SK-N-BE(2), SH-SY5Y) using real-time PCR and fluorescence in situ hybridization (FISH). Functional assays encompassing both overexpression and knockdown experiments in NB cell lines, as well as in vivo investigations, were conducted. RNA-seq analysis revealed a correlation between circ-SHPRH and the pathway of P21 (CDKN1A), a pivotal cell cycle regulator. Validation through PCR and other techniques confirmed that circ-SHPRH upregulated P21 expression. Furthermore, the regulatory role of circ-SHPRH in the P21-CDK pathway was corroborated through SHPRH-146aa expression analysis. Notably, adenovirus-mediated circ-SHPRH overexpression effectively curbed NB tumor growth in NSG mice, while combining circ-SHPRH with everolimus exhibited potential for NB treatment. This study elucidates the remarkable significance of circ-SHPRH in NB and its prospective utility in gene therapy, thereby paving the way for innovative therapeutic approaches.