Increased gene dosage of RFWD2 causes autistic-like behaviors and aberrant synaptic formation and function in mice.

Autism spectrum disorder (ASD) is a neurodevelopmental disorder characterized by impaired social interactions, communication deficits and repetitive behaviors. A study of autistic human subjects has identified RFWD2 as a susceptibility gene for autism, and autistic patients have 3 copies of the RFWD2 gene. The role of RFWD2 as an E3 ligase in neuronal functions, and its contribution to the pathophysiology of ASD, remain unknown. We generated RFWD2 knockin mice to model the human autistic condition of high gene dosage of RFWD2. We found that heterozygous knockin (Rfwd2+/-) male mice exhibited the core symptoms of autism. Rfwd2+/- male mice showed deficits in social interaction and communication, increased repetitive and anxiety-like behavior, and spatial memory deficits, whereas Rfwd2+/- female mice showed subtle deficits in social communication and spatial memory but were normal in anxiety-like, repetitive, and social behaviors. These autistic-like behaviors in males were accompanied by a reduction in dendritic spine density and abnormal synaptic function on layer II/III pyramidal neurons in the prelimbic area of the medial prefrontal cortex (mPFC), as well as decreased expression of synaptic proteins. Impaired social behaviors in Rfwd2+/- male mice were rescued by the expression of ETV5, one of the major substrates of RFWD2, in the mPFC. These findings indicate an important role of RFWD2 in the pathogenesis of autism.

Influence of Spider Silk Protein Structure on Mechanical and Biological Properties for Energetic Material Detection.

Spider silk protein, renowned for its excellent mechanical properties, biodegradability, chemical stability, and low immune and inflammatory response activation, consists of a core domain with a repeat sequence and non-repeating sequences at the N-terminal and C-terminal. In this review, we focus on the relationship between the silk structure and its mechanical properties, exploring the potential applications of spider silk materials in the detection of energetic materials.

Application of QSAR models for acute toxicity of tetrazole compounds administrated orally and intraperitoneally in rat and mouse.

Tetrazoles and their derivatives possess various biological activities, such as antibacterial, anti-fungal, and other activities. However, these compounds may induce specific cumulative and toxic effects in living organisms. Therefore, quantitative structure-activity relationship (QSAR) models were constructed to study the acute oral toxicity of tetrazoles in rats and mice. The toxicity data of 111 tetrazole compounds were collected using the ChemIDplus, ChEMBL and ECHA databases as response variables, while the PaDEL-descriptor generated the 2D descriptors as independent variables. The models were developed and validated following the OECD guidelines by the DTC-QSAR tool. Three QSAR models were successfully established for the oral routes of rat and mouse and the intraperitoneal route of mouse, respectively. The scatter plots showed high consistency between the training and test data sets. All the models successfully met the external and internal validation criteria. Most of the descriptors kept in the final models exhibited positive correlations with toxicity, whereas only 6 descriptors exhibited negative associations. Several chemicals were identified as response or structural outliers, based on the standardized residuals and leverage values. In conclusion, the findings of this investigation demonstrate that the proposed QSAR models hold promise in forecasting the acute toxicity of recently developed or synthesized tetrazole compounds, thereby mitigating potential risks to human health and the environment.

Salidroside protect Chinese hamster V79 cells from genotoxicity and oxidative stress induced by CL-20.

Hexanitrohexaazaisowurtzitane (CL-20) is a high-energy elemental explosive widely used in chemical and military fields. CL-20 harms environmental fate, biosafety, and occupational health. However, there is little known about the genotoxicity of CL-20, in particular its molecular mechanisms. Therefore, this study was framed to investigate the genotoxic mechanisms of CL-20 in V79 cells and evaluate whether the genotoxicity could be diminished by pretreating the cells with salidroside. The results showed that CL-20-induced genotoxicity in V79 cells primarily through oxidative damage to DNA and mitochondrial DNA (mtDNA) mutation. Salidroside could significantly reduce the inhibitory effect of CL-20 on the growth of V79 cells and reduce the levels of reactive oxygen species (ROS), 8-hydroxy-2 deoxyguanosine (8-OHdG), and malondialdehyde (MDA). Salidroside also restored CL-20-induced superoxide dismutase (SOD) and glutathione (GSH) in V79 cells. As a result, salidroside attenuated the DNA damage and mutations induced by CL-20. In conclusion, oxidative stress may be involved in CL-20-induced genotoxicity in V79 cells. Salidroside could protect V79 cells from oxidative damage induced by CL-20, mechanism of which may be related to scavenging intracellular ROS and increasing the expression of proteins that can promote the activity of intracellular antioxidant enzymes. The present study for the mechanisms and protection of CL-20-mediated genotoxicity will help further to understand the toxic effects of CL-20 and provide information on the therapeutic effect of salidroside in CL-20-induced genotoxicity.

Traumatic vertebra and endplate fractures promote adjacent disc degeneration: evidence from a clinical MR follow-up study.

OBJECTIVES: The integrity of endplate is important for maintaining the health of adjacent disc and trabeculae. Yet, pathological impacts of traumatic vertebra and endplate fractures were less studied using clinical approaches. This study aims to investigate their effects on the development of adjacent disc degeneration, segmental kyphosis, Modic changes (MCs), and high-intensity zones (HIZs). MATERIALS AND METHODS: Magnetic resonance (MR) images of patients with acute traumatic vertebral compression fractures (T11-L5) were studied. On MR images, endplate fractures were evaluated as present or absent. Disc signal, height, bulging area, sagittal Cobb angle, MCs, and HIZs were measured on baseline and follow-up MR images to study the changes of the disc in relation to vertebra fractures and endplate fractures. RESULTS: Ninety-seven patients were followed up for 15.4 ± 14.0 months. There were 123 fractured vertebrae, including 79 (64.2%) with endplate fractures and 44 (35.8%) without. Both the adjacent and control discs decreased in signal and height over time (p < 0.001), and the disc adjacent to vertebral fractures had greater signal and height loss than the control disc (p < 0.05). In the presence of endplate fractures, the adjacent discs had greater signal decrease in follow-up (p < 0.05), as compared to those without endplate fractures. Sagittal Cobb angle significantly increased in segments with endplate fractures (p < 0.05). Vertebra fractures were associated with new occurrence of MCs in the fractured vertebra (p < 0.001) but not HIZs in the adjacent disc. CONCLUSIONS: Traumatic vertebral fractures were associated with accelerated adjacent disc degeneration, which appears to be further promoted by concomitant endplate fractures. Endplate fractures were associated with progression of segmental kyphosis.

Sesamol supplementation alleviates nonalcoholic steatohepatitis and atherosclerosis in high-fat, high carbohydrate and high-cholesterol diet-fed rats.

Sesamol, a major ingredient in sesame seeds (Sesamum indicum L.) and its oil, is considered a powerful functional food ingredient. However, few studies have investigated its effects on high-fat, high carbohydrate and high-cholesterol (HF-HCC) diet-induced nonalcoholic steatohepatitis (NASH) complicated with atherosclerosis. The present study elucidates the protective effects of sesamol against NASH and atherosclerosis in HF-HCC diet-fed rats. Sprague-Dawley rats were supplemented with or without sesamol in drinking water (0.05 mg mL-1, 0.1 mg mL-1 and 0.2 mg mL-1) from the beginning to end. At the end of the experiment, sesamol supplementation suppressed HF-HCC diet-induced body weight gain and increased absolute liver and adipose tissue weights in rats. Serum biochemical analyses showed that sesamol supplementation improved HF-HCC diet-induced metabolism disorders and damaged vascular endothelial function. Histological examinations displayed that dietary sesamol not only alleviated hepatic balloon degeneration, steatosis, inflammation and fibrosis, but also mitigated lipid accumulation and fibrous elements in the aorta arch in HF-HCC diet-fed rats. In addition, sesamol supplementation inhibited hepatic NOD-like receptor protein 3 (NLRP3) expression and ERS-IRE1 signaling pathway activation. Moreover, sesamol treatment decreased uric acid levels both in serum and the liver by its effect on the inhibition of xanthine oxidase (XO) activity and/or its expression, which might be closely associated with the inhibitions of NLRP3 expression and ERS-IRE1 signaling pathway activation in HF-HCC diet-fed rats. These findings demonstrated that sesamol alleviated NASH and atherosclerosis in HF-HCC diet-fed rats, and may be a potent dietary supplement for protection against these diseases.

A Deep-Learning-Based, Fully Automated Program to Segment and Quantify Major Spinal Components on Axial Lumbar Spine Magnetic Resonance Images.

OBJECTIVE: The paraspinal muscles have been extensively studied on axial lumbar magnetic resonance imaging (MRI) for better understanding of back pain; however, the acquisition of measurements mainly relies on manual segmentation, which is time consuming. The study objective was to develop and validate a deep-learning-based program for automated acquisition of quantitative measurements for major lumbar spine components on axial lumbar MRIs, the paraspinal muscles in particular. METHODS: This study used a cross-sectional observational design. From the Hangzhou Lumbar Spine Study, T2-weighted axial MRIs at the L4-5 disk level of 120 participants (aged 54.8 years [SD = 15.0]) were selected to develop the deep-learning-based program Spine Explorer (Tulong). Another 30 axial lumbar MRIs were automatically measured by Spine Explorer and then manually measured using ImageJ to acquire quantitative size and compositional measurements for bilateral multifidus, erector spinae, and psoas muscles; the disk; and the spinal canal. Intersection-over-union and Dice score were used to evaluate the performance of automated segmentation. Intraclass coefficients and Bland-Altman plots were used to examine intersoftware agreements for various measurements. RESULTS: After training, Spine Explorer (Tulong) measures an axial lumbar MRI in 1 second. The intersections-over-union were 83.3% to 88.4% for the paraspinal muscles and 92.2% and 82.1% for the disk and spinal canal, respectively. For various size and compositional measurements of paraspinal muscles, Spine Explorer (Tulong) was in good agreement with ImageJ (intraclass coefficient = 0.85 to approximately 0.99). CONCLUSION: Spine Explorer (Tulong) is automated, efficient, and reliable in acquiring quantitative measurements for the paraspinal muscles, the disk, and the canal, and various size and compositional measurements were simultaneously obtained for the lumbar paraspinal muscles. IMPACT: Such an automated program might encourage further epidemiological studies of the lumbar paraspinal muscle degeneration and enhance paraspinal muscle assessment in clinical practice.

Changes of the adjacent discs and vertebrae in patients with osteoporotic vertebral compression fractures treated with or without bone cement augmentation.

BACKGROUND CONTEXT: Although vertebral augmentation with bone cement has been commonly used to treat symptomatic osteoporotic vertebral compression fractures, relatively little is known about the impact of augmentation on the adjacent spinal components. PURPOSE: To determine the imaging effects of vertebral augmentation on the adjacent discs, the augmented vertebra, and the involved spinal segment. STUDY DESIGN: Retrospective radiographic study. PATIENT SAMPLE: Patients with acute osteoporotic vertebral compression fractures who underwent vertebral augmentation or nonoperative treatments. OUTCOME MEASURES: On baseline and follow-up mid-sagittal T2W magnetic resonance images, quantitative measurements of disc degeneration, including disc height, bulging, and signal, vertebral height, wedge angle, and segmental kyphotic angle were acquired. METHODS: Lumbar spine magnetic resonance images of patients with acute osteoporotic vertebral compression fractures at a local hospital in Eastern China between 2010 and 2017 were reviewed. Student's t-tests and χ2 tests were used to examine the differences of baseline and changes over time between vertebrae underwent vertebral augmentation and those did not. Paired t-tests were used to examine the differences between baseline and follow-up to study the changes of adjacent disc degeneration, creep deformity of the vertebra and progression of segmental kyphosis. RESULTS: There were 112 acute vertebral compression fractures (72 treated with kyphoplasty and 40 with nonoperative treatments) in 101 subjects. At final follow-up (mean 21.5 months), the cranial disc of the augmented vertebra decreased in height (p<.001), and both cranial and caudal discs decreased in signal intensity (p≤.02). The discs in the nonoperative group did not undergo such degenerative changes. For the fractured vertebra, vertebral height significantly decreased (p<.01 for both) and vertebral wedge angle significantly increased (p≤.01 for both), regardless of augmentation treatment or not. Segmental kyphotic angle significantly increased in vertebral fractures that underwent vertebral augmentation (p<.001), but not in those underwent nonoperative treatments. CONCLUSIONS: Patients that underwent vertebral augmentation had more advanced disc degeneration at adjacent disc levels as compared to those without augmentation. The fractured vertebral body height decreased and the wedge angle increased, regardless of vertebral augmentation treatment or not. Vertebral augmentation may be associated with increased creep deformity of the adjacent vertebra and the progression of segmental kyphosis.

Spine Explorer: a deep learning based fully automated program for efficient and reliable quantifications of the vertebrae and discs on sagittal lumbar spine MR images.

BACKGROUND CONTEXT: Although quantitative measurements improve the assessment of disc degeneration, acquirement of quantitative measurements relies on manual segmentation on lumbar magnetic resonance images (MRIs), which may introduce subjective bias. To date, only a few semiautomatic systems have been developed to quantify important components on MRIs. PURPOSE: To develop a deep learning based program (Spine Explorer) for automated segmentation and quantification of the vertebrae and intervertebral discs on lumbar spine MRIs. STUDY DESIGN: Cross-sectional study. PATIENT SAMPLE: The study was extended on the Hangzhou Lumbar Spine Study, a population-based study of mainland Chinese with focuses on lumbar degenerative changes. From this population-based database, 50 sets lumbar MRIs were randomly selected as training dataset, and another 50 as test dataset. OUTCOME MEASURES: Regions of vertebrae and discs were manually segmented on T2W sagittal MRIs to train a convolutional neural network for automated segmentation. Intersection-over-union was calculated to evaluate segmentation performance. Computational definitions were proposed to acquire quantitative morphometric and signal measurements for lumbar vertebrae and discs. MRIs in the test dataset were automatically measured with Spine Explorer and manually with ImageJ. METHODS: Intraclass correlation coefficient (ICC) were calculated to examine inter-software agreements. Correlations between disc measurements and Pfirrmann score as well as age were examined to assess measurement validity. RESULTS: The trained Spine Explorer automatically segments and measures a lumbar MRI in half a second, with mean Intersection-over-union of 94.7% and 92.6% for the vertebra and disc, respectively. For both vertebra and disc measurements acquired with Spine Explorer and ImageJ, the agreements were excellent (ICC=0.81~1.00). Disc measurements significantly correlated to Pfirrmann score, and greater age was associated with greater anterior disc bulging area (r=0.35~0.44) and fewer signal measurements (r=-0.62~-0.77) as automatically acquired with Spine Explorer. CONCLUSIONS: Spine Explorer is an efficient, accurate, and reliable tool to acquire comprehensive quantitative measurements for lumbar vertebra and disc. Implication of such deep learning based program can facilitate clinical studies of the lumbar spine.

Entrapment of a metal foreign body in the cervical spinal canal during surgical procedure: A case report.

RATIONALE: Retention of foreign objects in spinal canal usually results from penetrating spinal trauma or failed internal instruments. However, entrapment of a foreign body in cervical spinal canal during surgery is rare, and whether such an object may cause neurological complications remains unknown in literature. PATIENT CONCERNS: A 50-year-old man underwent C5 corpectomy and instrumentation surgery due to cervical myelopathy. During the surgery, the cutting edge of a Kerrison rongeur was broken and the metal tip was retained behind C4 vertebra. DIAGNOSIS: Retention of foreign body in the cervical spinal canal. INTERVENTIONS: To remove the metal object, multiple strategies were tried but all failed. As such a metal object was thought to be dangerous to the spinal cord, a remedy C4 corpectomy was performed to remove it. Accidentally, however, the metal fragment further migrated to C2/3 canal. At last, the metal fragment had to be retained in the cervical spinal canal. OUTCOMES: At 2-year follow-up, the metal fragment remained in situ and no delayed complications occurred. LESSONS: We reported a rare case of metal object retention in cervical spinal canal due to rongeur fatigue fractures. Under certain circumstances, retention of a small foreign object in spinal canal may not lead to neurological complications. If failed to remove an entrapped foreign body, it may be safe to leave it in the spinal canal for further observation.

The circadian clock gene PER2 plays an important role in tumor suppression through regulating tumor-associated genes in human oral squamous cell carcinoma.

Low expression of the clock gene PER2 is closely related to carcinogenesis and the development of cancer; however, the mechanism of the low expression of PER2 that led to cell malignant transformation remains unclear. This study used RNA interference (RNAi) technology to silence PER2 in SCC15 human oral squamous cell carcinoma (OSCC) cells. Then it was found that the ability of cancer cell proliferation, migration, and invasion were markedly increased (P<0.05), and the ability of cancer cell apoptosis and the number of cells in G1/G0 phase were markedly reduced (P<0.05) after PER2 knockdown. PER2 knockdown increased the expression of Ki-67, MDM2, c-Myc, Bcl-2, MMP2, and VEGF mRNA (P<0.05), and decreased the expression of p53, Bax, and TIMP-2 mRNA (P<0.05). The in vivo experiments also proved that the tumorigenicity of SCC15 cells was significantly enhanced after PER2 silence (P<0.05). Overall, these results show that PER2 through the regulation of the numerous important downstream tumor-related genes, plays a major role in tumor suppression, and it may be a novel molecular target for cancer treatment.

The clock gene PER1 plays an important role in regulating the clock gene network in human oral squamous cell carcinoma cells.

The various clock genes in normal cells, through their interaction, establish a number of positive and negative feedback loops that compose a network structure. These genes play an important role in regulating normal physiological activities. The expression of clock gene PER1 is decreased in many types of cancer. PER1 is highly correlated with the initiation and progression of cancer by regulating numerous downstream genes. However, it is still unclear whether the lower expression of PER1 in cancer can influence the expression of other clock genes in the clock gene network. In this study, we used short hairpin RNA interference to knock down PER1 effectively in SCC15 human oral squamous cell carcinoma cells. These cancer cells later were subcutaneously injected into the back of nude mice. We discovered that after PER1 knockdown, apoptosis was decreased and cell proliferation and in vivo tumor formation were enhanced. Quantitative real-time PCR result indicated that in vitro and in vivo cancer cells after PER1 knockdown, PER2, DEC1, DEC2, CRY1, CRY2 and NPAS2 were significantly down-regulated at the mRNA level, while PER3, TIM, RORα and REV-ERBα were significantly up-regulated. It prompts that the role of PER1 in carcinogenesis is exerted not only by regulating downstream genes, but also through the synergistic dysregulation of many other clock genes in the clock gene network.

In-vivo imaging of oral squamous cell carcinoma by EGFR monoclonal antibody conjugated near-infrared quantum dots in mice.

OBJECTIVES: The purpose of this study was to investigate in-vivo visible imaging of oral squamous cell carcinoma (OSCC) by targeting epidermal growth factor receptor (EGFR) with near-infrared quantum dots. MATERIALS AND METHODS: Quantum dots with an emission wavelength of 800 nm (QD800) were conjugated to monoclonal antibodies against EGFR, resulting in the probe designated as QD800-EGFR Ab. OSCC cell line (BcaCD885) expressing high levels of EGFR was transplanted subcutaneously into nude mice cheeks to develop an OSCC animal model. QD800-EGFR Ab containing 100 pmol equivalent of QD800 was intravenously injected into the animal model, and in-situ and in-vivo imaging of cheek squamous cell carcinoma was analyzed at 10 different time points. RESULTS AND CONCLUSION: In-vivo imaging and immunohistochemical examination of the tumors showed that intravenously injected QD800-EGFR Ab probe could bind EGFR expressed on BcaCD885 cells. Fluorescence signals of BcaCD885 cells labeled with QD800-EGFR Ab probe could be clearly detected, and these fluorescence signals lasted for 24 hours. The most complete tumor images with maximal signal-to-noise ratio were observed from 15 minutes to 6 hours after injection of the probe. To the best of the authors' knowledge, this is the first study that has obtained clear in-situ and in-vivo imaging of head and neck cancer by using QD800-EGFR Ab probe. The authors conclude that the combination of near-infrared quantum dots that are highly penetrating for tissues with EGFR monoclonal antibody has promising prospects in in-vivo imaging of OSCC and development of personalized surgical therapies.

Screening the pathogenic genes and pathways related to DMBA (7,12-dimethylbenz[a]anthracene)-induced transformation of hamster oral mucosa from precancerous lesions to squamous cell carcinoma.

This study aimed to screen the pathogenic genes and pathways that relate to the transformation of hamster buccal mucosa from precancerous lesions to squamous cell carcinoma by whole genome microarray and bioinformatics analysis. A DMBA (7,12-dimethylbenz[a]anthracene)-induced hamster model of a precancerous lesion and squamous cell carcinoma was established. The differentially expressed genes were detected using an Agilent whole rat genome microarray, which contains 41,000 genes/ESTs. Gene ontology (GO) functional classification and pathway analyses were performed, and a subset of differentially expressed genes were validated using RT-PCR. The results showed that during the transformation of hamster buccal mucosa from the precancerous lesion to squamous cell carcinoma, a total of 1,981 genes were differentially expressed, of which 1,037 were up-regulated and 944 were down-regulated. GO analysis revealed that the differentially expressed genes are mainly involved in 14 functional groups including those of metabolism and cell structure. Additionally, 9 significantly altered pathways were identified. Among the 1,861 known differentially expressed genes, 14 genes including Casp3, CCL5 and CXCL12 were enriched in the 9 altered pathways. The up-regulation of SPARC and down-regulation of Casp3 were confirmed by RT-PCR. In conclusion, a total of 1,981 differentially expressed genes and 9 significantly altered pathways were identified in the transformation of hamster buccal mucosa from precancerous lesions to squamous cell carcinoma. A total of 14 pathway-enriched genes including Casp3, CCL5 and CXCL12 may play critical roles in the alteration of cellular pathways leading to the transformation of buccal mucosa from precancerous lesions to squamous cell carcinoma. Future studies focusing on these genes and pathways are required in order to gain a better understanding and provide effective prevention and treatment of oral squamous cell carcinoma.