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Osteosarcoma (OS) is the most common primary malignant pediatric bone tumor and is characterized by high heterogeneity. Studies have revealed a wide range of phenotypic differences among OS cell lines in terms of their in vivo tumorigenicity and in vitro colony-forming abilities. However, the underlying molecular mechanism of these discrepancies remains unclear. The potential role of mechanotransduction in tumorigenicity is of particular interest. To this end, we tested the tumorigenicity and anoikis resistance of OS cell lines both in vitro and in vivo. We utilized a sphere culture model, a soft agar assay, and soft and rigid hydrogel surface culture models to investigate the function of rigidity sensing in the tumorigenicity of OS cells. Additionally, we quantified the expression of sensor proteins, including four kinases and seven cytoskeletal proteins, in OS cell lines. The upstream core transcription factors of rigidity-sensing proteins were further investigated. We detected anoikis resistance in transformed OS cells. The mechanosensing function of transformed OS cells was also impaired, with general downregulation of rigidity-sensing components. We identified toggling between normal and transformed growth based on the expression pattern of rigidity-sensing proteins in OS cells. We further uncovered a novel TP53 mutation (R156P) in transformed OS cells, which acquired gain of function to inhibit rigidity sensing, thus sustaining transformed growth. Our findings suggest a fundamental role of rigidity-sensing components in OS tumorigenicity as mechanotransduction elements through which cells can sense their physical microenvironment. In addition, the gain of function of mutant TP53 appears to serve as an executor for such malignant programs.

SummaryNeural activity is increasingly recognized as a critical regulator of cancer growth. In the brain, neuronal activity robustly influences glioma growth both through paracrine mechanisms and through electrochemical integration of malignant cells into neural circuitry via neuron-to-glioma synapses, while perisynaptic neurotransmitter signaling drives breast cancer brain metastasis growth. Outside of the CNS, innervation of tumors such as prostate, breast, pancreatic and gastrointestinal cancers by peripheral nerves similarly regulates cancer progression. However, the extent to which the nervous system regulates lung cancer progression, either in the lung or when metastatic to brain, is largely unexplored. Small cell lung cancer (SCLC) is a lethal high-grade neuroendocrine tumor that exhibits a strong propensity to metastasize to the brain. Here we demonstrate that, similar to glioma, metastatic SCLC cells in the brain co-opt neuronal activity-regulated mechanisms to stimulate growth and progression. Optogenetic stimulation of cortical neuronal activity drives proliferation and invasion of SCLC brain metastases. In the brain, SCLC cells exhibit electrical currents and consequent calcium transients in response to neuronal activity, and direct SCLC cell membrane depolarization is sufficient to promote the growth of SCLC tumors. In the lung, vagus nerve transection markedly inhibits primary lung tumor formation, progression and metastasis, highlighting a critical role for innervation in overall SCLC initiation and progression. Taken together, these studies illustrate that neuronal activity plays a crucial role in dictating SCLC pathogenesis in both primary and metastatic sites.

Background Schwann cells (SCs) respond to nerve injury by transforming into the repair-related cell phenotype, which can provide the essential signals and spatial cues to promote axonal regeneration and induce target reinnervation. Endothelial cells (ECs) contribute to intraneural angiogenesis contributing to creating a permissive microenvironment. The coordination between ECs and SCs within injury sites is crucial in the regeneration process, however, it still unclear. As the intercellular vital information mediators in the nervous system, exosomes have been proposed to take a significant role in regulating regeneration. Thus, the main purpose of this study is to determine the facilitative effect of ECs-derived exosomes on SCs and to seek the underlying mechanism. Results In the present study, we collected exosomes from media of ECs. We demonstrated that exosomes derived from ECs possessed the favorable neuronal affinity both in vitro and in vivo. Further research indicated that EC-exosomes (EC-EXO) could boost and maintain repair-related phenotypes of SCs, thereby enhancing axonal regeneration, myelination of regenerated axons and neurologically functional recovery of the injured nerve. MiRNA sequencing in EXO-treated SCs and control SCs indicated that EC-EXO significantly up-regulated expression of miR199-5p. Furthermore, this study demonstrated that EC-EXO drove the conversion of SC phenotypes in a PI3K/AKT/PTEN-dependent manner. Conclusion In conclusion, our research indicates that the internalization of EC-EXO in SCs can promote nerve regeneration by boosting and maintaining the repair-related phenotypes of SCs. And the mechanism may be relevant to the up-regulated expression of miR199-5p and activation of PI3K/AKT/PTEN signaling pathway. Graphical Abstract

Disagreements over various unanswered questions about contribution of the adsorption process and functional groups on dye adsorption still exist. The main aim of this research was to evaluate the contributions of physical hole structure, hydrogen bond, and electrostatic attraction on dye adsorption. Three ideal representatives, namely, a sponge with porous structure, P(AM) containing -CONH2 groups, and P(AANa/AM) containing -COONa groups, were chosen to evaluate the above contributions. The methylene blue (MB) removal rates of these three products were compared through individual experiments. The results revealed that physical hole structure did not play a role in decreasing dye concentration. Hydrogen bond existed in dye adsorption but did not remarkably reduce dye concentration. The excellent removal results of P(AANa/AM) demonstrated that electrostatic attraction was critical in enriching dye contaminants from the solution into solid adsorbent. The results could provide insights into the dye adsorption mechanisms for further research.

Ferroptosis is a non-apoptotic regulated cell death caused by iron accumulation and subsequent lipid peroxidation. Currently, the therapeutic role of ferroptosis on cancer is gaining increasing interest. Baicalin an active component in Scutellaria baicalensis Georgi with anticancer potential various cancer types; however, the effects of baicalein on bladder cancer and the underlying molecular mechanisms remain largely unknown. In the study, we investigated the effect of baicalin on bladder cancer cells 5637 and KU-19-19. As a result, we show baicalin exerted its anticancer activity by inducing apoptosis and cell death in bladder cancer cells. Subsequently, we for the first time demonstrate baicalin-induced ferroptotic cell death in vitro and in vivo, accompanied by reactive oxygen species (ROS) accumulation and intracellular chelate iron enrichment. The ferroptosis inhibitor deferoxamine but not necrostatin-1, chloroquine (CQ), N-acetyl-l-cysteine, l-glutathione reduced, or carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]-fluoromethylketone (Z-VAD-FMK) rescued baicalin-induced cell death, indicating ferroptosis contributed to baicalin-induced cell death. Mechanistically, we show that ferritin heavy chain 1 (FTH1) was a key determinant for baicalin-induced ferroptosis. Overexpression of FTH1 abrogated the anticancer effects of baicalin in both 5637 and KU19-19 cells. Taken together, our data for the first time suggest that the natural product baicalin exerts its anticancer activity by inducing FTH1-dependent ferroptosis, which will hopefully provide a prospective compound for bladder cancer treatment., Graphical abstract Baicalin exerts its anticancer activity in bladder cancer by inducing ferritin heavy chain 1 (FTH1)-dependent ferroptosis, which will hopefully provide great therapeutic potential for bladder cancer treatment.Image 1

Increasing studies have proved that microRNAs (miRNAs) are critical biomarkers in the development of human complex diseases. Identifying disease-related miRNAs is beneficial to disease prevention, diagnosis and remedy. Based on the assumption that similar miRNAs tend to associate with similar diseases, various computational methods have been developed to predict novel miRNA-disease associations (MDAs). However, selecting proper features for similarity calculation is a challenging task because of data deficiencies in biomedical science. In this study, we propose a deep learning-based computational method named MAGCN to predict potential MDAs without using any similarity measurements. Our method predicts novel MDAs based on known lncRNA–miRNA interactions via graph convolution networks with multichannel attention mechanism and convolutional neural network combiner. Extensive experiments show that the average area under the receiver operating characteristic values obtained by our method under 2-fold, 5-fold and 10-fold cross-validations are 0.8994, 0.9032 and 0.9044, respectively. When compared with five state-of-the-art methods, MAGCN shows improvement in terms of prediction accuracy. In addition, we conduct case studies on three diseases to discover their related miRNAs, and find that all the top 50 predictions for all the three diseases have been supported by established databases. The comprehensive results demonstrate that our method is a reliable tool in detecting new disease-related miRNAs.

Background: Most prior studieshave used conventional statistical techniquessuch asthe nomogram to examine patient prognosis and survival based on the distinct subtypes and primary locations of chondrosarcoma. Nevertheless, few studies have examined overall survival in patients with non-metastatic chondrosarcomas. We aim to develop a machine learning algorithm to evaluate 5-year survival in this cohort and transform the most accurate prediction model into an online calculator for clinical use. Methods: Between 1975 and 2018, we gathered data on patients with non-metastatic chondrosarcoma from the Surveillance, Epidemiological, and End Result databases. From these data, nine features were used to develop four machine learning models, including the boosted decision tree, support vector machine, bayes point machine, and neural network models, which were then evaluated and compared based on discrimination, calibration, and overall performance. Results: Ultimately, 1202 patients met our inclusion criteria. With a c statistic of 0.88, a calibration slope of 0.997, a calibration intercept of -0.02, and a Brier score of 0.12, the Bayes point machine performed best overall, and it was integrated into a free, publicly available software interface that can be found at https://bayesglm.shinyapps.io/non-metastatic-chondrosarcoma/. Conclusion: Although this Bayes point machine-based prediction model has not been externally validated, the online calculator can be used as a reference tool for medical staff as well as patients on survival prediction. External validation of this prediction model should be the focus of future research to improve its credibility.

There is a need to standardize the process of micro/nanobubble preparation to bring it closer to clinical translation. We explored a neural network-based model to predict the structure-echogenicity relationship for the preparation and fabrication of ultrasound-enhanced contrast agents. Seven formulations were screened, and 109 measurements were obtained. An artificial neural network-multilayer perceptron (ANN-MLP) model was used. The original data were divided into the training and testing groups, which included 73 and 36 groups of data, respectively. The hidden layer was selected from three hidden layers and included bias. The classification graph showed that the predicted values of the training and testing groups were 76.7% and 66.7%, respectively. According to the receiver operating characteristic curve, the accuracy of different imaging effects could achieve a prediction rate of 88.1–96.5%. The percentage graph showed that the data were gradually converging. The predictive analysis curves of different ultrasound effects gradually approached stable value of Gain. Normalized importance predicted contributions for the Pk1, poly-dispersity index (PDI), and intensity account were 100%, 98.5%, and 89.7%, respectively. The application of the ANN-MLP model is feasible and effective for the exploration of the synthesis process of ultrasound contrast agents. 1,2-Distearoyl-sn-glycero-3 phosphoethanolamine-N (methoxy[polyethylene glycol]-2000) (DSPE PEG-2000) correlated highly with the success rate of contrast agent synthesis.

Within the basal ganglia, striatal dopamine (DA) and acetylcholine (Ach) are essential for the selection and reinforcement of motor actions and decision making. In vitro studies have revealed a circuit local to the striatum by which each of these two neurotransmitters directly regulates release of the other. Ach, released by a unique population of cholinergic interneurons (CINs), drives DA release via direct axonal depolarization. In turn, DA inhibits CIN activity via dopamine D2 receptors (D2R). Whether and how this circuit contributes to striatal function in vivo remains unknown. To define the in vivo role of this circuit, we monitored Ach and DA signals in the ventrolateral striatum of mice performing a reward-based decision-making task. We establish that DA and Ach exhibit multiphasic and anticorrelated transients that are modulated by decision history and reward outcome. However, CIN perturbations reveal that DA dynamics and reward-prediction error encoding do not require Ach release by CINs. On the other hand, CIN-specific deletion of D2Rs shows that DA inhibits Ach levels in a D2R-dependent manner, and loss of this regulation impairs decision-making. To determine how other inputs to striatum shape Ach signals, we assessed the contribution of projections from cortex and thalamus and found that glutamate release from both sources is required for Ach release. Altogether, we uncover a dynamic relationship between DA and Ach during decision making and reveal modes of CIN regulation by local DA signals and long-range cortical and thalamic inputs. These findings deepen our understanding of the neurochemical basis of decision making and behavior.

Background: Knee osteoarthritis (KOA) is a chronic and degenerative bone and joint disease, with knee osteoarthritis, cartilage degeneration, destruction and subchondral bone remodeling as the main pathological features. Its clinical symptoms are knee pain, swelling, limited activity, and long course of disease can cause joint deformities. At present, the early treatment of western medicine is mainly the use of non-steroidal drugs for anti-inflammation and removing pain, but because the efficacy of these drugs is unstable, the disease is easy to repeat after treatment, and the clinical effect is not good .Although Biqi capsule has advantages in the treatment of knee osteoarthritis, there is a lack of standard clinical studies to verify it, so the purpose of this randomized controlled study is to evaluate the efficacy and safety of Biqi capsule in the treatment of knee osteoarthritis. Methods: This is a prospective randomized controlled trial to study the efficacy and safety of Biqi capsule in the treatment of knee osteoarthritis. The patients were randomly divided into a treatment group and a control group according to 1:1. Among them, treatment group: Biqi capsule combined with diclofenac sodium sustained release tablets; Control group: Diclofenac sodium sustained-release tablets alone. Both groups were treated with standard treatment for 2 weeks and were followed up for 30 days to pay attention to the efficacy and safety indexes. Observation indicators included: the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), Hospital for Special Surgery Knee Score (HSS), liver and kidney function, adverse reactions and so on. SPSS 25.0 software is used for data analysis. Discussion: This study will evaluate the efficacy and safety of Biqi capsule in the treatment of knee osteoarthritis, and the results of this experiment will provide a clinical basis for Biqi capsule in the treatment of knee osteoarthritis.

BackgroundIncreasing biomedical studies have shown that the dysfunction of miRNAs is closely related with many human diseases. Identifying disease-associated miRNAs would contribute to the understanding of pathological mechanisms of diseases. Supervised learning-based computational methods have continuously been developed for miRNA-disease association predictions. Negative samples of experimentally-validated uncorrelated miRNA-disease pairs are required for these approaches, while they are not available due to lack of biomedical research interest. Existing methods mainly choose negative samples from the unlabelled ones randomly. Therefore, the selection of more reliable negative samples is of great importance for these methods to achieve satisfactory prediction results.ResultsIn this study, we propose a computational method termed as KR-NSSM which integrates two semi-supervised algorithms to select more reliable negative samples for miRNA-disease association predictions. Our method uses a refinedK-means algorithm for preliminary screening of likely negative and positive miRNA-disease samples. ARocchioclassification-based method is applied for further screening to receive more reliable negative and positive samples. We implement ablation tests in KR-NSSM and find that the combination of the two selection procedures would obtain more reliable negative samples for miRNA-disease association predictions. Comprehensive experiments based on fivefold cross-validations demonstrate improvements in prediction accuracy on six classic classifiers and five known miRNA-disease association prediction models when using negative samples chose by our method than by previous negative sample selection strategies. Moreover, 469 out of 1123 selected positive miRNA-disease associations by our method are confirmed by existing databases.ConclusionsOur experiments show that KR-NSSM can screen out more reliable negative samples from the unlabelled ones, which greatly improves the performance of supervised machine learning methods in miRNA-disease association predictions. We expect that KR-NSSM would be a useful tool in negative sample selection in biomedical research.

Background: Schwann cells respond to nerve injury by transforming into a cell phenotype that promotes repair. These repair Schwann cells provide the essential signals and spatial cues to encourage axonal regeneration and induce target reinnervation. Endothelial cells (ECs), another crucial component of nerve tissue, contribute to the formation of regenerative microenvironment. Therefore, the way to control these peculiar cells, amplify their supportive functions and prevent their deterioration could be remarkable for the treatment of injured peripheral nerves. As the vital information mediators in the nervous system, exosomes have been proposed to take a significant role in regulating regeneration.Results: In the present study, we collected exosomes from media of ECs. We demonstrated that exosomes derived from ECs possessed the favorable neuronal affinity both in vitro and in vivo. Further research indicated that EC-exosomes could boost and maintain repair-related phenotypes of Schwann cells, thereby enhancing axonal regeneration, myelination of regenerated axons and neurologically functional recovery of the injured nerve. We found some highly enriched miRNAs in EC-exosomes, including let-7f-5p, miR199-5p and miR-3074-5p by the sequencing of miRNAs. In addition, miRNA sequencing in EXO-treated Schwann cells and control Schwann cells indicated that EC-exosomes significantly up-regulated the expression of miR199-5p. Furthermore, this study demonstrated that EC-exosomes drove the conversion of Schwann cell phenotypes in a PI3K/AKT/PTEN-dependent manner. Conclusion: In conclusion, our research indicates that the internalization of EC-exosomes in Schwann cells can promote nerve regeneration by boosting and maintaining the repair-related phenotypes of Schwann cells. And the mechanism may be relevant to the regulation of miRNAs and activation of PI3K/AKT/PTEN signaling pathway.

Increasing biomedical evidence has proved that the dysregulation of miRNAs is associated with human complex diseases. Identification of disease-related miRNAs is of great importance for disease prevention, diagnosis and remedy. To reduce the time and cost of biomedical experiments, there is a strong incentive to develop efficient computational methods to infer potential miRNA-disease associations. Although many computational approaches have been proposed to address this issue, the prediction accuracy needs to be further improved. In this study, we present a computational framework MKGAT to predict possible associations between miRNAs and diseases through graph attention networks (GATs) using dual Laplacian regularized least squares. We use GATs to learn embeddings of miRNAs and diseases on each layer from initial input features of known miRNA-disease associations, intra-miRNA similarities and intra-disease similarities. We then calculate kernel matrices of miRNAs and diseases based on Gaussian interaction profile (GIP) with the learned embeddings. We further fuse the kernel matrices of each layer and initial similarities with attention mechanism. Dual Laplacian regularized least squares are finally applied for new miRNA-disease association predictions with the fused miRNA and disease kernels. Compared with six state-of-the-art methods by 5-fold cross-validations, our method MKGAT receives the highest AUROC value of 0.9627 and AUPR value of 0.7372. We use MKGAT to predict related miRNAs for three cancers and discover that all the top 50 predicted results in the three diseases are confirmed by existing databases. The excellent performance indicates that MKGAT would be a useful computational tool for revealing disease-related miRNAs.

In order to analyze the running state of rolling bearing, the principle component analysis is used to extract new parameters to analyze the running state of the rolling bearing. Firstly, the whole life cycle vibration signal of the rolling bearing is collected by using an acceleration sensor. Secondly, Empirical Mode Decomposition (EMD) is used to reduce the noise in the vibration signal. Then, the main components of the time domain parameters are extracted using the principle component of the denoised signal. And the main components are synthesized into new parameter indicators. Finally, under the new parameter index, the envelope analysis is used to analyze the signal classification. The results show that the new parameter index can divide the entire life cycle of the rolling bearing into four different operating periods, each representing a different bearing operating state.

Peripheral nerves have complex and precise structures that differ from other types of tissues and intrinsic regeneration abilities after injury. Spontaneous recovery is possible for neuropraxia and axonotmesis, while surgical treatment is required for neurotmesis. It remains a challenge to repair nerve gaps, a series of severe neurotmesis. It seems that 3 cm is the upper limit distance for primate peripheral nerves to regenerate spontaneously. Nerve autografts are the gold standard treatment for bridging nerve gaps. In the present review, current biomaterials for repairing gaps after peripheral nerve injury are briefly summarized. Moreover, the microstructure of the peripheral nerve, classifications of peripheral nerve injury, and the Wallerian degeneration are reviewed in the biological view and clinical practice. The failure of nerve regeneration in nerve conduits bridging longer than 3 cm gaps may be contributing to the insufficient vascularization of nerve conduit materials. Future researchers could focus on advanced biomaterials that promoting the angiogenesis of nerve conduits.

The production connection is a crucial component of the Inter City Association. An urban network based on the enterprise perspective better reflects the structural characteristics of regional cities. Based on data gathered from the headquarters and branches of the listed manufacturing enterprises in 2020, this paper analyzes the county-level administrative units in the Beijing-Tianjin-Hebei region of China. Using the subordinate connection model and the social network analysis method, this paper examines the spatial structural characteristics and factors of urban networks in the Beijing-Tianjin-Hebei region. The results suggest that resource allocation in the Beijing-Tianjin-Hebei region is unbalanced, with a significant difference in urban radiation and agglomeration capacity. As the administrative centers, Beijing, Tianjin, and Shijiazhuang show a strong ability to allocate resources within the network. The overall network density in the region was shown to be relatively low, with the main links being of low or medium level. The urban network is defined by the network connection led by Beijing and Tianjin with Beijing-Tianjin-Tangshan and Beijing-Baoding-Shijiazhuang as the main axis. These cities exhibit a "dense southeast and sparse northwest" pattern. From a hierarchical perspective, high-level network connections are based mainly on spatial proximity. Analyzing the agglomerative subgroups, the study found that the inner and outer connections of the core subgroups were relatively high. Semi-marginal subgroups generally accepted the radiation of core subgroups, while marginal subgroups had little connection with other subgroups in the region. The results of the QAP analysis show that the administrative relationship, spatial distance, city size difference, economic development level difference, industrial structure difference, and labor cost difference have a significant influence on the urban network of the Beijing-Tianjin-Hebei region.

Objective To compare the pedicle morphology of nondystrophic scoliosis secondary to neurofibromatosis type 1 (ND-NF1) scoliosis secondary to neurofibromatosis type 1 (NF1-S) and adolescent idiopathic scoliosis (AIS) using computed tomography (CT) in a consistent classification system. Methods The transverse pedicle widths of cancellous and cortical channels were measured using preoperative spinal CT images (T1–L5) and then classified as having type A–E pedicles; type B, C, D, and E pedicles were regarded as abnormal. A subset of these patients, a postoperative CT was performed to evaluate the accuracy of pedicle screw placement. According to the perforation of the medial or lateral pedicle walls by the pedicle screw, the placement of screw was classified by a grade ranging from 0 to 3, and grades 2 and 3 were regarded as misplacement. Results In total, 3230 pedicles were measured and classified, and the incidence rate of abnormal pedicles in the ND-NF1 group was significantly greater than that of the AIS group (65.9% vs. 61.4%, P = 0.03). The misplacement rate in the ND-NF1 group was significantly greater than that in the AIS group (12.2% vs. 7.4%; P = 0.01), and in terms of the corresponding 1100 pedicles, the incidence rate of abnormal pedicles in the ND-NF1 group was higher than that in the AIS group (71.7% vs. 64.9%; P = 0.02). Conclusions The incidence rate of abnormal pedicles in patients with ND-NF1 is significantly greater than that of patients with AIS, and it may the reason for the higher misplacement rate of pedicle screws in patients with ND-NF1.

Multiparametric magnetic resonance imaging and targeted biopsy have been widely accepted as the most accurate technique to detect localize prostate cancer. It is a time-consuming and expensive option and may not be widely available in China, making ultrasound the first choice for the detection of prostate cancer. In this current retrospective study, the diagnostic values of enhanced transrectal ultrasound, contrast-enhanced transrectal ultrasound, and real-time sonoelastography were evaluated. Symptomatic 315 men older than 40 years with prostate-specific antigen level greater than 4.0 ng/mL, with abnormal digital rectal examinations, and with suspicious lesions for prostate cancer under enhanced transrectal ultrasound included in the study. Enhanced transrectal ultrasound was suspicious in all 315 men, with 189 of 315 men with prostate cancer according to the prostate biopsy report. Sonoelastography was suspicious in 294 of 315 men, with 166 of 315 men with prostate cancer according to the prostate biopsy report. Contrast-enhanced transrectal ultrasound was suspicious in 221 of 315 men, with 159 of 315 men with prostate cancer according to the prostate biopsy report. Real-time sonoelastography alone and contrast-enhanced transrectal ultrasound alone were missed in 27 (11%) and 39 (15%) lesions to report cancer through biopsies. Working area for enhanced transrectal ultrasound, real-time sonoelastography, and contrast-enhanced transrectal ultrasound for detection of prostate cancer were 0 to 1 diagnostic confidence, 0.11 to 0.895 diagnostic confidence, and 0.39 to 0.63 diagnostic confidence respectively. Real-time sonoelastography and contrast-enhanced transrectal ultrasound may improve the detection of prostate cancer in men with suspicious prostate lesions under enhanced transrectal ultrasound (Level of Evidence: III; Technical Efficacy Stage: 2).

The atherosclerotic plaque is characterized by narrowing of blood vessels and reduced blood flow leading to the insufficient blood supply to the brain. The hemodynamic changes caused by arterial stenosis increase the shearing force of the fibrous cap on the surface of the plaque, thereby reducing the stability of the plaque. Unstable plaques are more likely to promote angiogenesis and increase the risk of patients with cerebrovascular diseases. A timely understanding of the formation and stability of the arterial plaque can guide in taking targeted measures for reducing the risk of acute stroke in patients. It has been confirmed that nano-microbubbles can enter these plaques through the gaps in the patient’s vascular endothelial cells, thereby enhancing the acquisition of ultrasound information for plaque visualization. Therefore, we aim to investigate the diagnostic value of targeted nano-microbubbles for atherosclerotic plaques. This study constructed vascular endothelial growth factor receptor-2 (VEGFR-2) targeting antibody nano-microbubbles and compared its diagnostic value with that of blank nano-microbubbles for atherosclerotic plaques. Studies have found that VEGFR-2 targeting antibody nano-microbubbles can accurately detect the position of plaques. Its detection rate, sensitivity, and specificity for plaques are higher than those of blank nano-microbubbles. Similarly, the peak intensity and average transit time of VEGFR-2 targeting antibody nano-microbubbles were greater than those of blank nano-microbubbles. Therefore, we believe that the combination of VEGFR-2 antibody and nano-microbubbles can enhance the acquisition of ultrasound information on atherosclerotic plaque neovascularization, thereby improving the early diagnosis of unstable plaque.

Objective. In this paper, we study the role of the VHL gene in regulating the proliferation and apoptosis of renal cell carcinoma, as well as the safety and transfection efficiency of ultrasound microbubble gene transfection technology. Method. We use kidney cancer cell lines as an in vitro research object and apply ultrasound microbubble gene transfection technology to transfect the VHL gene into kidney cancer cell line (786-0). The proliferation and apoptosis of cells were measured to clarify the inhibitory effect of the VHL gene in renal cell carcinoma. After that, pEGFP-VHL was transfected using ultrasonic microbubble and liposome gene transfection techniques, respectively, and the transfection efficiency was measured by immunofluorescence. Results. Compared with untreated and 786-0 cells that are transfected with empty vector, the expression level of VHL gene mRNA in 786-0 cells that are transfected with pcDNA3.1-VHL was significantly increased, and the cell growth inhibition rate was significantly higher. The rate of apoptosis increased significantly. Transfection efficiency of the pEGFP-VHL gene after transfection of 786-0 cells for 48 h: control group 0, liposome group ( 35.55 ± 2.77 ) %, ultrasound microbubble group ( 18.27 ± 2.83 ) %, and two transfection methods on cells. There is no significant difference in the impact of vitality. Conclusion. VHL gene expression can significantly inhibit the proliferation ability of renal cancer cell line 786-0 and promote its apoptosis. VHL gene is a potential target for gene therapy of kidney cancer.

To our best knowledge, no study investigated the utility of the combination of high-frequency US and real-time shear wave elastography (RTSWE) in the evaluation of Achilles tendon (AT). Our study aimed to evaluate AT in patients with diabetic foot ulcers using high-frequency US and RTSWE. We retrospectively reviewed 100 patients who visited our hospital due to diabetes. Patients were divided into the study group (those with diabetic foot ulcers) and the control group (those without diabetic foot ulcers). Patients’ demographics, basic medical records, and laboratory tests were reviewed. High-frequency ultrasound and RTSWE were performed in both AT for all 100 patients. Young’s modulus was measured at the upper, middle, and lower parts of each AT by RTSWE. There were 50 patients with diabetic foot ulcers. Patients in the study group had older age, a higher incidence of insulin use, and a higher level of cholesterol than those in the control group. However, thickness and Young’s modulus of AT were comparable in two groups, when evaluated by the combination of high-frequency ultrasound and RTSWE. There was no significant difference in thickness and elastic modulus of AT in patients with and without diabetic foot ulcers when evaluated by the combination of high-frequency ultrasound and RTSWE.

Objective: In this study, we aimed to develop an amino-terminal fragment (ATF) peptide-targeted liposome carrying β-elemene (ATF24-PEG-Lipo-β-E) for targeted delivery into urokinase plasminogen activator receptor-overexpressing bladder cancer cells combined with cisplatin (DDP) for bladder cancer treatment. Methods: The liposomes were prepared by ethanol injection and high-pressure microjet homogenization. The liposomes were characterized, and the drug content, entrapment efficiency, and in vitro release were studied. The targeting efficiency was investigated using confocal microscopy, ultra-fast liquid chromatography, and an orthotopic bladder cancer model. The effects of ATF24-PEG-Lipo-β-E combined with DDP on cell viability and proliferation were evaluated by a Cell Counting Kit-8 (CCK-8) assay, a colony formation assay, and cell apoptosis and cell cycle analyses. The anticancer effects were evaluated in a KU-19-19 bladder cancer xenograft model. Results: ATF24-PEG-Lipo-β-E had small and uniform sizes (˜79 nm), high drug loading capacity (˜5.24 mg/mL), high entrapment efficiency (98.37 ± 0.95%), and exhibited sustained drug release behavior. ATF24-PEG-Lipo-β-E had better targeting efficiency and higher cytotoxicity than polyethylene glycol (PEG)ylated β-elemene liposomes (PEG-Lipo-β-E). DDP, combined with ATF24-PEG-Lipo-β-E, exerted a synergistic effect on cellular apoptosis and cell arrest at the G2/M phase, and these effects were dependent on the caspase-dependent pathway and Cdc25C/Cdc2/cyclin B1 pathways. Furthermore, the in vivo antitumor activity showed that the targeted liposomes effectively inhibited the growth of tumors, using the combined strategy. Conclusions: The present study provided an effective strategy for the targeted delivery of β-elemene (β-E) to bladder cancer, and a combined strategy for bladder cancer treatment.

Background and Purpose: RAS mutations limit the effectiveness of anti-epidermal growth factor receptor (EGFR) monoclonal antibodies in combination with chemotherapy for metastatic colorectal cancer (mCRC) patients. Therefore, new cell death forms have focused on identifying indirect targets to inhibit Ras-induced oncogenesis. Recently, emerging evidence has shown the potential of triggering ferroptosis for cancer therapy, particularly for eradicating aggressive malignancies that are resistant to traditional therapies. Methods: KRAS mutant CRC cell HCT116 and Lovo were treated with cetuximab and β-elemene, a bioactive compound isolated from Chinese herb Curcumae Rhizoma. Ferroptosis and epithelial-mesenchymal transformation (EMT) were detected in vitro and in vivo. Orthotopic CRC animal model were established and the tumor growth was monitored by IVIS bioluminescence imaging. Tumor tissues were collected to determine ferroptosis effect and the expression of EMT markers after the treatment. Results: CCK-8 assay showed that synergetic effect was obtained when 125 µg/ml β-elemene was combined with 25 µg/ml cetuximab in KRAS mutant CRC cells. AV/PI staining suggested a non-apoptotic mode of cell death after the treatment with β-elemene and cetuximab. In vitro, β-elemene in combination with cetuximab was shown to induce iron-dependent reactive oxygen species (ROS) accumulation, glutathione (GSH) depletion, lipid peroxidation, upregulation of HO-1 and transferrin, and downregulation of negative regulatory proteins for ferroptosis (GPX4, SLC7A11, FTH1, glutaminase, and SLC40A1) in KRAS mutant CRC cells. Meanwhile, combinative treatment of β-elemene and cetuximab inhibited cell migration and decreased the expression of mesenchymal markers (Vimentin, N-cadherin, Slug, Snail and MMP-9), but promoted the expression of epithelial marker E-cadherin. Moreover, ferroptosis inhibitors but not other cell death suppressors abrogated the effect of β-elemene in combination with cetuximab on KRAS mutant CRC cells. In vivo, co-treatment with β-elemene and cetuximab inhibited KRAS mutant tumor growth and lymph nodes metastases. Conclusions: Our data for the first time suggest that the natural product β-elemene is a new ferroptosis inducer and combinative treatment of β-elemene and cetuximab is sensitive to KRAS mutant CRC cells by inducing ferroptosis and inhibiting EMT, which will hopefully provide a prospective strategy for CRC patients with RAS mutations.

Objective: In this study, we aimed to develop an amino-terminal fragment (ATF) peptide-targeted liposome carrying β-elemene (ATF24-PEG-Lipo-β-E) for targeted delivery into urokinase plasminogen activator receptor-overexpressing bladder cancer cells combined with cisplatin (DDP) for bladder cancer treatment. Methods: The liposomes were prepared by ethanol injection and high-pressure microjet homogenization. The liposomes were characterized, and the drug content, entrapment efficiency, and in vitro release were studied. The targeting efficiency was investigated using confocal microscopy, ultra-fast liquid chromatography, and an orthotopic bladder cancer model. The effects of ATF24-PEG-Lipo-β-E combined with DDP on cell viability and proliferation were evaluated by a Cell Counting Kit-8 (CCK-8) assay, a colony formation assay, and cell apoptosis and cell cycle analyses. The anticancer effects were evaluated in a KU-19-19 bladder cancer xenograft model. Results: ATF24-PEG-Lipo-β-E had small and uniform sizes (˜79 nm), high drug loading capacity (˜5.24 mg/mL), high entrapment efficiency (98.37 ± 0.95%), and exhibited sustained drug release behavior. ATF24-PEG-Lipo-β-E had better targeting efficiency and higher cytotoxicity than polyethylene glycol (PEG)ylated β-elemene liposomes (PEG-Lipo-β-E). DDP, combined with ATF24-PEG-Lipo-β-E, exerted a synergistic effect on cellular apoptosis and cell arrest at the G2/M phase, and these effects were dependent on the caspase-dependent pathway and Cdc25C/Cdc2/cyclin B1 pathways. Furthermore, the in vivo antitumor activity showed that the targeted liposomes effectively inhibited the growth of tumors, using the combined strategy. Conclusions: The present study provided an effective strategy for the targeted delivery of β-elemene (β-E) to bladder cancer, and a combined strategy for bladder cancer treatment.

Purpose: To investigate the correlation between the expression of PD-L1, SOCS3 and immune-related biomarkers CD276, CD4, CD8 in hepatocellular carcinoma (HCC) and further determine the relationship with clinicopathologic characteristics and the prognostic value of their co-expression in HCC patients. Methods: We assessed the expression of PD-L1, CD276, SOCS3, CD4 and CD8 by immunohistochemistry in tumor tissue from 74 HCC patients who underwent curative hepatectomy. Results: High expression of PD-L1 was significantly associated with high Edmondson grade (p0.05). High expression of CD8 was respectively significantly associated with worse overall survival (OS) (p=0.002). There was no significantly difference between CD4 and CD8 high-expression and overall survival (OS) (p=0.100). Both high expression of PD-L1 (p=0.003) and low expression of SOCS3 (p=0.015) was significantly associated with worse overall survival (OS). But CD276 only had a trendency (p=0.166). Additionally, multivariate Cox regression models implied that PD-L1, SOCS3, as well as both CD4 and CD8 was an independent prognostic factor for OS (p

Background: Traditional Chinese medicine (TCM) syndrome, also named syndrome, are comprehensive and integral analyses of clinical information which helps to guide different individualized treatment prescriptions. Methods: Thirty healthy controls and 80 colorectal cancer (CRC) patients (including 33 Spleen Qi Deficiency syndrome, 23 Dampness heat Heat syndrome, 17 Blood stasis Stasis syndrome and 7 other syndrome) were enrolled into this study. Human mRNAs were extracted from peripheral blood mononuclear cells. The gene expression for CRC patients with different TCM syndrome was determined by microarray and qRT-PCR. Results: Spleen Qi Deficiency, Dampness Heat and Blood Stasis were the most common syndromes in CRC patients. There is a significant difference was found in mRNA expression levels (especially for PIK3CA, STAT3, SOX9 and KDM5C) among Spleen Qi Deficiency, Dampness Heat and Blood Stasis syndrome groups. The higher mRNA levels of JNK1, TP53, MLH1, MSH6, PMS2, SOCS3, TCF7L2, FAM123B, PSAP, FBXW7, SALL4 and the lower expression of inflammatory cytokine IL-6 were found in Spleen Qi Deficiency group but not other syndrome types. The higher mRNA levels of KRAS, MUC16, EGFR, GRASP65, PIK3CA, MAPK7, CD24, STAT3, SLC11A1, Bcl-2, TXNDC17 and some inflammatory cytokines (IL-6, IL-23, TNF-a, CXCR4) were found in Dampness Heat group but not other syndrome types. Blood Stasis syndrome showed higher expression of SOX9, MLH1, MSH6, KDM5C, PCDH11X, PSAP and SALL4, and lower mRNA levels of PIK3CA, CD24, STAT3, CXCR4, TXNDC17 and TP53. The CRC patients with Dampness Heat syndrome may might have a poor prognosis than other syndrome types. Conclusion: The identification of syndrome conditions had different impacts on CRC prognosis, and which may might be related with different mRNA expression levels. Some oncogenes and pro-inflammatory cytokines were highly expressed in Dampness Heat group but not other syndrome types, suggesting that the CRC patients with Dampness Heat syndrome may might have a poor prognosis. Our results prelimitarily uncovered the molecular basis of syndrome differences in CRC prognosis, a better understanding for TCM treatment of CRC.

5-Fluorouracil (5-FU) is known as a first-line chemotherapeutic agent against colorectal cancer (CRC), but drug resistance occurs frequently and significantly limits its clinical success. Our previous study showed that the protocadherin 17 (PCDH17) gene was frequently methylated and functioned as a tumor suppressor in CRC. However, the relationship between PCDH17 and 5-FU resistance in CRC remains unclear. Here, we revealed that PCDH17 was more highly expressed in 5-FU-sensitive CRC tissues than in 5-FU-resistant CRC tissues, and high expression of PCDH17 was correlated with high BECN1 expression. Moreover, this expression profile contributed to superior prognosis and increased survival in CRC patients. Restoring PCDH17 expression augmented the 5-FU sensitivity of CRC in vitro and in vivo by promoting apoptosis and autophagic cell death. Furthermore, autophagy played a dominant role in PCDH17-induced cell death, as an autophagy inhibitor blocked cell death to a greater extent than the pancaspase inhibitor Z-VAD-FMK. PCDH17 inhibition by siRNA decreased the autophagy response and 5-FU sensitivity. Mechanistically, we showed that c-Jun NH2-terminal kinase (JNK) activation was a key determinant in PCDH17-induced autophagy. The compound SP600125, an inhibitor of JNK, suppressed autophagy and 5-FU-induced cell death in PCDH17-reexpressing CRC cells. Taken together, our findings suggest for the first time that PCDH17 increases the sensitivity of CRC to 5-FU treatment by inducing apoptosis and JNK-dependent autophagic cell death. PCDH17 may be a potential prognostic marker for predicting 5-FU sensitivity in CRC patients.

SUMMARY The neural substrates and pathophysiological mechanisms underlying the onset of cognitive and motor deficits in autism spectrum disorders (ASDs) remain unclear. Mutations in ASD-associated SHANK3 in mice (Shank3B−/−) result in the accelerated maturation of corticostriatal circuits during the second and third postnatal weeks. Here, we show that during this period, there is extensive remodeling of the striatal synaptic proteome and a developmental switch in glutamatergic synaptic plasticity induced by cortical hyperactivity in striatal spiny projection neurons (SPNs). Behavioral abnormalities in Shank3B−/− mice emerge during this stage and are ameliorated by normalizing excitatory synapse connectivity in medial striatal regions by the downregulation of PKA activity. These results suggest that the abnormal postnatal development of striatal circuits is implicated in the onset of behavioral deficits in Shank3B−/− mice and that modulation of postsynaptic PKA activity can be used to regulate corticostriatal drive in developing SPNs of mouse models of ASDs and other neurodevelopmental disorders., In Brief Peixoto et al. show that the onset of behavioral deficits in Shank3B−/− mice occurs during early postnatal development and that these can be ameliorated by reducing the glutamatergic synaptic drive in medial regions of the striatum by the downregulation of PKA activity., Graphical Abstract

Genetic diagnosis is a promising approach because several single-nucleotide polymorphisms (SNPs) associated with adolescent idiopathic scoliosis (AIS) progression have been reported. We review the predictive value of SNPs in curve progression of adolescent idiopathic scoliosis.We reviewed DNA-based prognostic testing to predict curve progression. Then, the multiple polymorphisms in loci related to AIS progression were also reviewed, and we elucidated the predictive value of SNPs from four functional perspectives, including endocrine metabolism, neuromuscular system, cartilage and extracellular matrix, enzymes, and cytokines.The ScoliScores were less successful predictors than expected, and the weak power of predictive SNPs might account for its failure. Susceptibility loci in ESR1, ESR2, GPER, and IGF1, which related to endocrine metabolism, have been reported to predict AIS progression. Neuromuscular imbalance might be a potential mechanism of scoliosis, and SNPs in LBX1, NTF3, and SOCS3 have been reported to predict the curve progression of AIS. Susceptibility loci in SOX9, MATN1, AJAP1, MMP9, and TIMP2, which are related to cartilage and extracellular matrix, are also potentially related to AIS progression. Enzymes and cytokines play essential roles in regulating bone metabolism and embryonic development. SNPs in BNC2, SLC39A8, TGFB1, IL-6, IL-17RC, and CHD7 were suggested as predictive loci for AIS curve progression.Many promising SNPs have been identified to predict the curve progression of AIS. However, conflicting results from replication studies and different ethnic groups hamper their reliability. Convincing SNPs from multiethnic populations and functional verification are needed.

Erianin, a natural product derived from Dendrobium chrysotoxum Lindl, has been proved to play antitumor activity in various cancers. However, the effects and molecular mechanisms of erianin in bladder cancer cells remain unexplored. In this study, we found that erianin triggered cell death and cell cycle arrest in bladder cancer cells. Then we demonstrated that erianin could promote the accumulation of lethal lipid-based reactive oxygen species (ROS) and the depletion of glutathione (GSH), suggesting the induction of ferroptosis. In the further study, the ferroptosis inhibitor deferoxamine (DFO), N-Acetylcysteine (NAC) and GSH but not necrostatin-1, CQ or Z-VAD-FMK rescued erianin-caused cell death, showing ferroptosis played a major role in erianin-caused cell death. In vivo, we also showed that erianin suppressed the tumor growth by inducing ferroptosis. Mechanistically, we demonstrated that nuclear factor E2-related factor 2 (NRF2) inactivation was a key determinant of ferroptosis caused by erianin. In bladder cancer cells, the compound tert-butylhydro-quinone (TBHQ), an activator of NRF2, suppressed erianin-induced ferroptosis. Whereas, NRF2 inhibition used shRNA augmented the ferroptosis response induced by erianin treatment. In conclusion, our data provide the first evidence that erianin can initiate ferroptosis-like cell death and lipid peroxidation in bladder cancer, which will hopefully become a promising anticancer compound for the treatment of bladder cancer.

Background: Chondrosarcoma is a rare type of bone tumor which more commonly found in adults range from 40 to 60 years old. Few studies has described the characteristic and prognostic factors of patients older than 60 years. This study aimed to study this feature and identify the prognostic factors based on SEER database.Methods: Thus, we collected clinicopathological data of chondrosarcoma patients in the Surveillance, Epidemiology, and End Results registry database from 1975 to 2018, and then use the Kaplan-Meier to analyze the patients’ survival. We also utilize Cox proportional hazard model to explore the prognostic factors and relevant characteristic including patients’ baseline demographics (age, race, and gender), tumor characteristics (tumor extension, histologic subtype, therapy, primary site, stage and grade.Results: After the implementation of exclusion criteria, there were 610 patients with chondrosarcoma older than 60 years. Our data showed that the incidence of chondrosarcoma is slightly higher in men than in women (52.3% vs 47.7%). In general, 90.8% of tumor had metastasized to distant sites. Meanwhile, 41.8% of tumors occurred in axial location (pelvis, spine, and ribs), 50.8% of tumors occurred in extremity (long or short bones of the upper or lower extremity), and 7.4% in other location (mandible, skull, and other atypical locations). Dedifferentiated chondrosarcoma (hazard ratio [HR] =2.553; 95% confidence interval [CI]= 1.754-3.716), grade (g2:HR:=1.299; 95% CI:=0.888-1.900, g3:HR=1.839;95% CI= 1.174-2.881, g4:HR=3.284,95%CI=2.053-5.253), distant metastasis (HR=3.264; 95% CI= (2.288-4.058), non-surgery perform (HR = 2.854; 95% CI= 2.022-4.028) were independent risk factors for 5-year overall survival.Conclusion: In conclusion, higher grade, non-surgery perform, dedifferentiated chondrosarcoma and distant metastasis indicated worse prognosis survival. Surgery can significantly improve the survival time of patients.

Based on ASP.NET framework, The Intelligent Estimated System for Rational Deployment of Medical Equipment (MERDIS) is designed and developed with SQL Server 2012 database and C# language. The system is used to realize the rational deployment suggestions and evaluation of medical equipment in hospitals. The system input the data of hospital medical equipment and clinical pathway into the database, and then feedback the deployment information to users which are calculated by big data information, so as to achieve the purpose of giving rational deployment of hospital medical equipment.

Colorectal cancer (CRC) is among the most prevalent cancers. Long non-coding RNAs (lncRNAs) are important participant in various cancers. Based on the literature, lncRNA RASSF8-AS1 inhibits laryngeal squamous cell carcinoma (LSCC) malignant progression. However, the role of RASSF8-AS1 in CRC remains unclear.This study centered on uncovering the role of RASSF8-AS1 and its related regulatory mechanisms in CRC cells.RT-qPCR and western blot were performed to examine the expression of target genes. Functional assays were conducted to determine the effect of target genes on the migration and invasion of CRC cells. Mechanism assays were also carried out to figure out the specific downstream mechanisms of RASSF8-AS1. In vivo assays were also involved.The expression of RASSF8-AS1 and RASSF8 was positively correlated in CRC, and the two genes were down-regulated in CRC cells and tissues. Moreover, CRC cell invasion and migration as well as xenograft CRC tumor growth suppressed by RASSF8-AS1 overexpression were entirely recovered by RASSF8 knockdown or partially rescued by miR-33a-5p augment. As for the downstream mechanism, RASSF8-AS1 sponged miR-33a-5p to up-regulate RASSF8, or recruited HNRNPC to stabilize RASSF8 mRNA.RASSF8-AS1 modulates miR-33a-5p/HNRNPC/RASSF8 axis to further impede CRC cell invasion and migration.The research data is confidential.

Neuron-glia interactions play a critical role in the regulation of synapse formation and circuit assembly. Here we demonstrate that canonical Sonic hedgehog (Shh) pathway signaling in cortical astrocytes acts to coordinate layer-specific synaptic connectivity. We show that the Shh receptor Ptch1 is expressed by cortical astrocytes during development and that Shh signaling is necessary and sufficient to promote the expression of genes involved in regulating synaptic development and layer-enriched astrocyte molecular identity. Loss of Shh in layer V neurons reduces astrocyte complexity and coverage by astrocytic processes in tripartite synapses; conversely, cell-autonomous activation of Shh signaling in astrocytes promotes cortical excitatory synapse formation. Furthermore, Shh-dependent genes Lrig1 and Sparc distinctively contribute to astrocyte morphology and synapse formation. Together, these results suggest that Shh secreted from deep-layer cortical neurons acts to specialize the molecular and functional features of astrocytes during development to shape circuit assembly and function.

Nutrient availability varies seasonally and spatially in the wild. The resulting nutrient limitation or restricted access to nutrients pose a major challenge for every organism. While many animals, such as hibernating animals, evolved strategies to overcome periods of nutrient scarcity, the cellular mechanisms of these strategies are poorly understood. Cave environments represent an extreme example of nutrient deprived environments since the lack of sunlight and therefore primary energy production drastically diminishes the nutrient availability. Here, we used Astyanax mexicanus, which includes river-dwelling surface fish and cave adapted cavefish populations to study the genetic adaptation to nutrient limitations. We show that cavefish populations store large amounts of fat in different body regions when fed ad libitum in the lab. We found higher expression of lipogenesis genes in cavefish livers when fed the same amount of food as surface fish, suggesting an improved ability of cavefish to use lipogenesis to convert available energy into triglycerides for storage into adipose tissue. Moreover, the lipid metabolism regulator, Peroxisome proliferator-activated receptor γ (Pparγ), is upregulated at both transcript and protein levels in cavefish livers. Chromatin Immunoprecipitation sequencing (ChIP seq) showed that Pparγ binds cavefish promoter regions of genes to a higher extent than surface fish. Finally, we identified two possible regulatory mechanisms of Pparγ in cavefish: higher amounts of ligands of the nuclear receptor, and nonsense mutations in per2, a known repressor of Pparγ. Taken together, our study reveals that upregulated Pparγ promotes higher levels of lipogenesis in the liver and contributes to higher body fat accumulation in cavefish populations, an important adaptation to nutrient limited environments.

Midbrain dopamine (mDA) neurons play a central role in reward signaling and are widely implicated in psychiatric and neurodegenerative disorders. To understand how mDA neurons perform these functions, it is important to understand how mDA-specific genes are regulated. However, cellular heterogeneity in the mammalian brain presents a major challenge to obtaining this understanding. To this end, we developed a virus-based approach to label and capture mDA nuclei for transcriptome (RNA-Seq), and low-input chromatin accessibility (liDNase-Seq) profiling, followed by predictive modeling to identify putative transcriptional regulators of mDA neurons. Using this method, we identified Gmeb1, a transcription factor predicted to regulate expression of Th and Dat, genes critical for dopamine synthesis and reuptake, respectively. Gmeb1 knockdown in mDA neurons resulted in downregulation of Th and Dat, as well as in severe motor deficits. This study thus identifies Gmeb1 as a master regulator of mDA gene expression and function, and provides a general method for identifying cell type-specific transcriptional regulators., Despite the known role of midbrain dopaminergic (mDA) signaling in the homeostatic control of mood and motor functions, little is known about how gene expression in these neurons is regulated. Here, authors develop an in vivo nuclear tagging and capture technique for low-input chromatin accessibility and transcriptome profiling of genetically-defined neuron populations to identify Gmeb1 as a novel transcriptional regulator of mDA neurons, whose loss of function impairs motor control in mice.

The purpose of this study was to evaluate computed tomography (CT) doses in child examinees at different ages throughout the Shanghai metropolitan area. The head and body CT dose indices (CTDIs) of 50 CT scanners were tested by phantom measurements using standard imaging protocols. The values of CTDIw, CTDIvol and dose length product (DLP) were calculated and saved in a table along with the parameters of routine head and chest scans for different age groups of children and adults. The effective doses were estimated from the K-factors by age and DLP. The CTDIvol, DLP and effective dose for multi-detector row CT (MDCT) in children during routine head scans were larger than those for single-detector row CT (SDCT) and dual-detector row CT (DDCT). The CTDIvol, DLP and effective dose for MDCT and DDCT in children during routine chest scans were lower than those for SDCT. Radiation risks are higher for children in CT examinations compared to adults.

Background: Cervical Spondylotic radiculopathy (CSR) is the most common spinal degenerative disease. Its clinical manifestations are pain and numbness in the neck and arm and limitation of neck movement, which greatly affects the life and work of patients. Acupuncture and electroacupuncture are commonly used in China, the efficacy of acupuncture has been confirmed. Existing evidence shows that electroacupuncture seems to be better than acupuncture, but there is a lack of clinical research to directly compare the two. Methods: This is a prospective randomized controlled trial to compare the efficacy of electroacupuncture and acupuncture in the treatment of CSR and to explore the safety and potential mechanism of electroacupuncture in the treatment of CSR. Approved by the Clinical Research Ethics Committee of our hospital, the patients are randomly divided into an experimental group (electroacupuncture group) or control group (acupuncture group). The patients are followed up for 30 days after 4 weeks of treatment. Observation indexes included VAS score, Neck Disability Index, Yasuhisa Tanaka 20 Score Scale, adverse reactions and so on. Finally, the data will be analyzed by SPSS 18.0 software. Discussion: This study will directly compare the advantages and disadvantages of electroacupuncture and acupuncture in the treatment of CSR. The results of this study will help to guide patients with CSR to choose appropriate treatment. Trial registration: OSF Registration number: DOI 10.17605/OSF.IO/9MKPN

The aim of the study was to determine if multiple intraspinal anomalies increase the risk of scoliosis correction compared to the normal intraspinal condition or 1 or 2 intraspinal anomalies in congenital scoliosis (CS) and whether correction for multiple intraspinal anomalies need to be performed with preliminary neurosurgical intervention before scoliosis correction. A total of 318 consecutive CS patients who underwent corrective surgery without preliminary neurosurgical intervention at a single institution from 2008 to 2016 were retrospectively reviewed, with a minimum of 2 years of follow-up. The patients were divided into 3 groups according to different intraspinal conditions. In the normal group (N group; n = 196), patients did not have intraspinal anomalies. In the abnormal group (A group; n = 93), patients had 1 or 2 intraspinal anomalies. In the multiple anomaly group (M group; n = 29), patients had 3 or more intraspinal anomalies including syringomyelia, split cord malformation [SCM], tethered cord, low conus, intraspinal mass, Chiari malformation or/and arachnoid cyst. The occurrence of complications as well as perioperative and radiographic data were analyzed. The incidence rate of multiple intraspinal anomalies in CS patients was 9.1% (29/318). No significant difference was observed in the perioperative outcomes or radiographic parameters at the final follow-up. There were no significant differences among the 3 groups in the total, major or neurological complication rates (all P > .05). Two patients (1 in the N group and 1 in the A group) experienced transient neurological complications, whereas no patient experienced permanent neurological deficits during surgery or follow-up. To our knowledge, the current study reported the largest cohort of intraspinal anomalies in patients with CS that has been reported in the literature. The results of our study demonstrated that patients with congenital scoliosis associated with intraspinal anomalies, even multiple intraspinal anomalies that coexist with more complex intraspinal pathologies, may safely and effectively achieve scoliosis correction without preliminary neurological intervention. More complex intraspinal pathologies do not seem to increase the risk of neurosurgical complications during corrective surgery.

Neuron-glia interactions play a critical role in the regulation of synapse formation and neuron specification. The cellular and molecular mechanisms by which neurons and astrocytes communicate and coordinate are not well understood. Here we demonstrate that canonical Sonic hedgehog (Shh) pathway signaling in cortical astrocytes acts to coordinate layer-specific synaptic connectivity and functional circuit development. We show that the Shh receptor Ptch1 is expressed by cortical astrocytes during development and that Shh signaling is necessary and sufficient to promote the expression of genes involved in regulating synaptic development and layer-specific astrocyte molecular identity. Loss of Shh in layer V neurons reduces astrocyte complexity and coverage by astrocytic processes in tripartite synapses, conversely, cell-autonomous activation of Shh signaling in astrocytes promotes cortical excitatory synapse formation. Furthermore, we determined that Shh-dependent genes Lrig1 and Sparc distinctively contribute to astrocyte morphology and synapse formation. Together, these results suggest that Shh secreted from deep layer cortical neurons acts to specialize the molecular and functional features of astrocytes during development to shape circuit assembly and function.

The release of urine, or micturition, serves a fundamental physiological function and, in many species, is critical for social communication. In mice, the pattern of urine release is modulated by external and internal factors and transmitted to the spinal cord via the pontine micturition center (PMC). Here, we exploited a behavioral paradigm in which mice, depending on strain, social experience, and sensory context, either vigorously cover an arena with small urine spots or deposit urine in a few isolated large spots. We refer to these micturition modes as, respectively, high and low territory-covering micturition (TCM) and find that the presence of a urine stimulus robustly induces high TCM in socially isolated mice. Comparison of the brain networks activated by social isolation and by urine stimuli to those upstream of the PMC identified the lateral hypothalamic area as a potential modulator of micturition modes. Indeed, chemogenetic manipulations of the lateral hypothalamus can switch micturition behavior between high and low TCM, overriding the influence of social experience and sensory context. Our results suggest that both inhibitory and excitatory signals arising from a network upstream of the PMC are integrated to determine context- and social-experience-dependent micturition patterns.

Neurons communicate by the activity-dependent release of small-molecule neurotransmitters packaged into synaptic vesicles (SVs). Although many molecules have been identified as neurotransmitters, technical limitations have precluded a full metabolomic analysis of synaptic vesicle content. Here, we present a workflow to rapidly isolate SVs and to interrogate their metabolic contents at a high-resolution using mass spectrometry. We validated the enrichment of glutamate in SVs of primary cortical neurons using targeted polar metabolomics. Unbiased and extensive global profiling of SVs isolated from these neurons revealed that the only detectable polar metabolites they contain are the established neurotransmitters glutamate and GABA. Finally, we adapted the approach to enable quick capture of SVs directly from brain tissue and determined the neurotransmitter profiles of diverse brain regions in a cell-type specific manner. The speed, robustness, and precision of this method to interrogate SV contents will facilitate novel insights into the chemical basis of neurotransmission.

Purpose To evaluate the relationship between the DNA mismatch repair (MMR) status and histone lysine‐specific demethylase 6A (KDM6A) on the prognosis of colorectal cancer (CRC). Methods About 594 patients with CRC from The Cancer Genome Atlas (TCGA) were enrolled in this retrospective study. Subsequently, a series of different classification tests for MMR status, cancer types, and target gene expression was conducted. Results After filtering out the KDMs group of genes, we selected KDM6A as the target gene. A significant difference in the performance of KDM6A in tumor and normal tissues were confirmed. Our results showed a lower KDM6A expression, lower KDM6A exon expression, and higher KDM6A DNA methylation than their corresponding normal tissues in colon adenocarcinoma (COAD). Notably, the main MMR genes were highly expressed in tumor tissues than normal tissues both in COAD and rectum adenocarcinoma (READ). Moreover, proficient DNA mismatch repair (pMMR) was found to be an important poor prognostic factor in COAD (p = 0.0064) and the low KDM6A expression was an important factor for poor prognosis in READ (p = 0.0217). Based on these results, we consequently relate MMR status with KDM6A expression in predicting the prognosis of patients with CRC. Moreover, patients with pMMR exhibited a low KDM6A expression in COAD (p = 0.0250). Samples were divided into two groups based on the KDM6A expression. Interestingly, the group with low KDM6A expression showed no difference between pMMR and deficient DNA mismatch repair (dMMR) in prognosis, whereas the group with high KDM6A expression was closely related to MMR status in OS (p = 0.0082). Besides, COAD patients with high KDM6A expression and pMMR status had poor OS (p = 0.0082). Conclusions The KDM6A/MMR classification‐based subtypes of low KDM6A expression/READ, high KDM6A expression/pMMR, and COAD/pMMR were associated with poor prognosis. This classification can be a novel prognostic approach in CRC., The KDM6A/MMR classification based subtypes of low KDM6A expression/READ, high KDM6A expression/pMMR, and COAD/pMMR were associated with poor prognosis. This classification could more accurately stratify CRC patients as per the prognostic performance.

SUMMARYNeuron-glia relationships play a critical role in the regulation of synapse formation and neuronal specification. The cellular and molecular mechanisms by which neurons and astrocytes communicate and coordinate are not well understood. Here we demonstrate that the canonical Sonic hedgehog (Shh) pathway is active in cortical astrocytes, where it acts to coordinate layer-specific synaptic connectivity and functional circuit development. We show that Ptch1 is a Shh receptor that is expressed by cortical astrocytes during development and that Shh signaling is necessary and sufficient to promote the expression of layer-specific astrocyte genes involved in regulating synapse formation and function. Loss of Shh in layer V neurons reduces astrocyte complexity and coverage by astrocytic processes in tripartite synapses, moreover, cell-autonomous activation of Shh signaling in astrocytes promotes cortical excitatory synapse formation. Together, these results suggest that Shh secreted from deep layer cortical neurons acts to specialize the molecular and functional features of astrocytes during development to shape circuit assembly and function.

The mouse cerebral cortex contains neurons that express choline acetyltransferase (ChAT) and are a potential local source of acetylcholine. However, the neurotransmitters released by cortical ChAT+ neurons and their synaptic connectivity are unknown. We show that the nearly all cortical ChAT+ neurons in mice are specialized VIP+ interneurons that release GABA strongly onto other inhibitory interneurons and acetylcholine sparsely onto layer 1 interneurons and other VIP+/ChAT+ interneurons. This differential transmission of ACh and GABA based on the postsynaptic target neuron is reflected in VIP+/ChAT+ interneuron pre-synaptic terminals, as quantitative molecular analysis shows that only a subset of these are specialized to release acetylcholine. In addition, we identify a separate, sparse population of non-VIP ChAT+ neurons in the medial prefrontal cortex with a distinct developmental origin that robustly release acetylcholine in layer 1. These results demonstrate both cortex-region heterogeneity in cortical ChAT+ interneurons and target-specific co-release of acetylcholine and GABA.