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Abstract Clinical studies have shown that there may be a certain relationship between pathological changes of the myodural bridge complex (MDBC) and chronic headaches of unknown cause. But there is still a lack of experimental evidence to explain the possible mechanism. This study aims to further confirm this relationship between MDBC and chronic headaches and explore its potential occurrence mechanism in rats. Bleomycin (BLM) or phosphate-buffered saline (PBS) was injected into the myodural bridge fibers of rats to establish the hyperplastic model of MDBC. After 4 weeks, the occurrence of headaches in rats was evaluated through behavioral scores. The immunohistochemistry staining method was applied to observe the expression levels of headache-related neurotransmitters in the brain. Masson trichrome staining results showed that the number of collagen fibers of MDBC was increased in the BLM group compared to those of the other two groups. It revealed hyperplastic changes of MDBC. The behavioral scores of the BLM group were significantly higher than those of the PBS group and the blank control group. Meanwhile, expression levels of CGRP and 5-HT in the headache-related nuclei of the brain were increased in the BLM group. The current study further confirms the view that there is a relationship between pathological changes of MDBC and chronic headaches of unknown cause. This study may provide anatomical and physiological explanations for the pathogenesis of some chronic headaches of unknown cause.

ABSTRACTLong non-coding RNAs (lncRNAs) have been shown to be involved in the regulation of skeletal muscle development through multiple mechanisms. The present study revealed that the lncRNA SOX6 AU (SRY-box transcription factor 6 antisense upstream) is reverse transcribed from upstream of the bovine sex-determining region Y (SRY)-related high-mobility-group box 6 (SOX6) gene. SOX6 AU was significantly differentially expressed in muscle tissue among different developmental stages in Xianan cattle. Subsequently, knockdown and overexpression experiments discovered that SOX6 AU promoted primary skeletal muscle cells proliferation, apoptosis, and differentiation in bovine. The overexpression of SOX6 AU in bovine primary skeletal muscle cells resulted in 483 differentially expressed genes (DEGs), including 224 upregulated DEGs and 259 downregulated DEGs. GO functional annotation analysis showed that muscle development-related biological processes such as muscle structure development and muscle cell proliferation were significantly enriched. KEGG pathway analysis revealed that the PI3K/AKT and MAPK signaling pathways were important pathways for DEG enrichment. Notably, we found that SOX6 AU inhibited the mRNA and protein expression levels of the SOX6 gene. Moreover, knockdown of the SOX6 gene promoted the proliferation and apoptosis of bovine primary skeletal muscle cells. Finally, we showed that SOX6 AU promoted the proliferation and apoptosis of bovine primary skeletal muscle cells by cis-modulation of SOX6 in cattle. This work illustrates our discovery of the molecular mechanisms underlying the regulation of SOX6 AU in the development of beef.

Background and objectivesGlioblastoma (GBM) and brain metastasis (MET) are the two most common intracranial tumors. However, the different pathogenesis of the two tumors leads to completely different treatment options. In terms of magnetic resonance imaging (MRI), GBM and MET are extremely similar, which makes differentiation by imaging extremely challenging. Therefore, this study explores an improved deep learning algorithm to assist in the differentiation of GBM and MET.Materials and methodsFor this study, axial contrast-enhanced T1 weight (ceT1W) MRI images from 321 cases of high-grade gliomas and solitary brain metastasis were collected. Among these, 251 out of 270 cases were selected for the experimental dataset (127 glioblastomas and 124 metastases), 207 cases were chosen as the training dataset, and 44 cases as the testing dataset. We designed a new deep learning algorithm called SCAT-inception (Spatial Convolutional Attention inception) and used five-fold cross-validation to verify the results.ResultsBy employing the newly designed SCAT-inception model to predict glioblastomas and brain metastasis, the prediction accuracy reached 92.3%, and the sensitivity and specificity reached 93.5 and 91.1%, respectively. On the external testing dataset, our model achieved an accuracy of 91.5%, which surpasses other model performances such as VGG, UNet, and GoogLeNet.ConclusionThis study demonstrated that the SCAT-inception architecture could extract more subtle features from ceT1W images, provide state-of-the-art performance in the differentiation of GBM and MET, and surpass most existing approaches.

Abstract The Myodural Bridge (MDB) is a physiological structure that is highly conserved in mammals and many of other tetrapods. It connects the suboccipital muscles to the cervical spinal dura mater (SDM) and transmits the tensile forces generated by the suboccipital muscles to the SDM. Consequently, the MDB has broader physiological potentials than just fixing the SDM. It has been proposed that MDB significantly contributes to the dynamics of cerebrospinal fluid (CSF) movements. Animal models of suboccipital muscle atrophy and hyperplasia were established utilizing local injection of BTX-A and ACE-031. In contrast, animal models with surgical severance of suboccipital muscles, and without any surgical operation were set as two types of negative control groups. CSF secretion and reabsorption rates were then measured for subsequent analysis. Our findings demonstrated a significant increase in CSF secretion rate in rats with the hyperplasia model, while there was a significant decrease in rats with the atrophy and severance groups. We observed an increase in CSF reabsorption rate in both the atrophy and hyperplasia groups, but no significant change was observed in the severance group. Additionally, our immunohistochemistry results revealed no significant change in the protein level of six selected choroid plexus-CSF-related proteins among all these groups. Therefore, it was indicated that alteration of MDB-transmitted tensile force resulted in changes of CSF secretion and reabsorption rates, suggesting the potential role that MDB may play during CSF circulation. This provides a unique research insight into CSF dynamics.

The taxonomic status of Gloydius shedaoensis qianshanensis Li, 1999 has long been debated due to its narrow distribution range, low population, loss of type specimens, and lack of detailed descriptions. Neotypes were designated based on newly collected specimens in the Qianshan Mountain area, detailed comparisons between G. s. shedaoensis, and G. s. qianshanensis were recorded, while we redescribed G. s. qianshanensis. PCA results based on the external morphology of the two subspecies of G. shedaoensis demonstrate the inter-population distinction of G. s. shedaoensis and G. s. qianshanensis, including head length, and dorsoventral-postorbital stripe width. Additionally, the ecological data of G. s. qianshanensis were recorded as supplements while providing further conservation strategies.

Background: West Nile virus is a severe zoonotic pathogen that can cause severe central nervous system symptoms in humans and horses, and is fatal for birds, chickens and other poultry. With no specific drugs or vaccines available, antibody-based therapy is a promising treatment. This study aims to develop neutralizing antibodies against West Nile virus and assess their cross-protective potential against Japanese encephalitis virus. Methods: Monoclonal antibodies against WNV and JEV were isolated by hybridoma technology. The therapeutic efficacy of these antibodies was evaluated using a mouse model, and a humanized version of the monoclonal antibody was generated for potential human application. Results: In this study, we generated eight monoclonal antibodies that exhibit neutralizing activity against WNV. Their therapeutic effects against WNV were validated both in vivo and in vitro. Among these antibodies, C9-G11-F3 also exhibited cross-protective activity against JEV. We also humanized the antibody to ensure that it could be used for WNV infection treatment in humans. Conclusion: This study highlights the importance of neutralizing antibodies as a promising approach for protection against West Nile virus infection and suggests their potential utility in the development of therapeutic interventions.

Abstract Myodural bridge (MDB) is a dense connective tissue between suboccipital muscle and dura mater. However, there are few reports on the development and maturation of the human MDB. This study aims to explore the developmental relationship between suboccipital muscle and MDB. 30 head and neck specimens from human fetuses (F) ranging from the 12th to 41st week (W) were made into histological sections. The F12W sections showed evidence that the dura mater dominated by fibroblasts, attached to the posterior atlanto-axial membrane (PAAM) which completely sealed the atlanto-axial space. In the F13W stage, myofibrils of the suboccipital muscle fibers increased significantly in number. At the F14W stage, a gap was observed at the caudal end of the PAAM. Numerous myodural bridge-like structures were observed blending into the dura mater through the gap. At the F19W stage, muscle cells mature. Starting at the F21W stage, the MDB were observed as fibroblasts that cross the atlanto-axial interspace and attach to the dura mater. Therefore, the traction generated by the suboccipital muscles seems to promote the maturity of MDB. This study will provide new morphological knowledge to support future research on the function of the human MDB and regulating the development mechanism of MDB.

Abstract The cisterna magna has been defined as the space between the inferior margin of the cerebellar vermis to the level of the foramen magnum, while an enlarged dorsal subarachnoid space at the occipito-cervical junction extending from the foramen magnum to the upper border of the axis (C2) is still ignored. Recently, the myodural bridge complex is proved to drive the cerebral spinal fluid flowing via this region, we therefore introduce the “occipito-atlantal cistern (OAC)” to better describe the subarachnoid space and provide a detailed rationale. The present study utilized several methods, including MRI, gross anatomical dissection, P45 sheet plastination, and three-dimensional visualization. OAC was observed to be an enlarge subarachnoid space, extending from the foramen magnum to the level of the C2. In the median sagittal plane, OAC was a funnel shape and its anteroposterior dimensions were 15.92 ± 4.20 mm at the level of the C0, 4.49 ± 1.25 mm at the level of the posterior arch of the C1, and 2.88 ± 0.77 mm at the level of the arch of the C2, respectively. In the median sagittal plane, the spino-dural angle of the OAC was calculated to be 35.10 ± 6.91°, and the area of OAC was calculated to be 232.28 ± 71.02 mm2. The present study provides OAC is a subarachnoid space independent from the cisterna magna. Because of its distinctive anatomy, as well as theoretical and clinical significance, OAC deserves its own name.

Pelophylax plancyi (Lataste, 1880), Pelophylax chosenicus (Okada, 1931) and Pelophylax hubeiensis (Fei & Ye, 1982) were described chronologically from East Asia. The three species have similar morphological and molecular characteristics, but the taxonomic relationships amongst them have long been ambiguous. To deal with this taxonomic issue, we examined newly-obtained topotypic specimens of P. plancyi, P. chosenicus and P. hubeiensis for morphological comparison. Furthermore, we investigated the phylogeny of pond frogs in Eurasia by Bayesian Inference and Maximum Likelihood analyses of a fragment consisting of mitochondrial DNA gene 16s and provided a molecular phylogeny of the genus Pelophylax. There were no morphological and molecular differences between P. plancyi and P. chosenicus, but both morphological and molecular differences between P. hubeiensis and P. plancyi. Hence, we conclude that P. chosenicus is a junior synonym of P. plancyi and P. hubeiensis is a separate species from P. plancyi.

African Swine Fever (ASF), caused by the African swine fever virus (ASFV), has inflicted significant economic losses on the pig industry in China. The key to mitigating its impact lies in accurate screening and strict biosecurity measures. In this regard, the development of colloidal gold immunochromatographic test strips (CGITS) has proven to be an effective method for detecting ASFV antibodies. These test strips are based on the ASFV p30 recombinant protein and corresponding monoclonal antibodies. The design of the test strip incorporates a high-concentration colloidal gold-labeled p30 recombinant protein as the detection sensor, utilizing Staphylococcal Protein A (SPA) as the test line (T line), and p30 monoclonal antibody as the control line (C line). The sensitivity and specificity of the test strip were evaluated after optimizing the labeling concentration, pH, and protein dosage. The research findings revealed that the optimal colloidal gold labeling concentration was 0.05 %, the optimal pH was 8.4, and the optimal protein dosage was 10 μg/mL. Under these conditions, the CGITS demonstrated a detection limit of 1:512 dilution of ASFV standard positive serum, without exhibiting cross-reactivity with antibodies against other viral pathogens. Furthermore, the test strips remained stable for up to 20 days when stored at 50 °C and 4 °C. Comparatively, the CGITS outperformed commercial ELISA kits, displaying a sensitivity of 90.9 % and a specificity of 96.2 %. Subsequently, 108 clinical sera were tested to assess its performance. The data showed that the coincidence rate between the CGITS and ELISA was 93.5 %. In conclusion, the rapid colloidal gold test strip provides an efficient and reliable screening tool for on-site clinical detection of ASF in China. Its accuracy, stability, and simplicity make it a valuable asset in combating the spread of ASF and limiting its impact on the pig industry.

Alzheimer's disease (AD) is the most common cause of dementia. Accurate prediction and diagnosis of AD and its prodromal stage, i.e., mild cognitive impairment (MCI), is essential for the possible delay and early treatment for the disease. In this paper, we adopt the data from the China Longitudinal Aging Study (CLAS), which was launched in 2011, and includes a joint effort of 15 institutions all over the country. Four thousand four hundred and eleven people who are at least 60 years old participated in the project, where 3,514 people completed the baseline survey. The survey collected data including demographic information, daily lifestyle, medical history, and routine physical examination. In particular, we employ ensemble learning and feature selection methods to develop an explainable prediction model for AD and MCI. Five feature selection methods and nine machine learning classifiers are applied for comparison to find the most dominant features on AD/MCI prediction. The resulting model achieves accuracy of 89.2%, sensitivity of 87.7%, and specificity of 90.7% for MCI prediction, and accuracy of 99.2%, sensitivity of 99.7%, and specificity of 98.7% for AD prediction. We further utilize the SHapley Additive exPlanations (SHAP) algorithm to visualize the specific contribution of each feature to AD/MCI prediction at both global and individual levels. Consequently, our model not only provides the prediction outcome, but also helps to understand the relationship between lifestyle/physical disease history and cognitive function, and enables clinicians to make appropriate recommendations for the elderly. Therefore, our approach provides a new perspective for the design of a computer-aided diagnosis system for AD and MCI, and has potential high clinical application value.

A novel wideband quasi-Chebyshev bandpass filter (BPF) with steep skirt selectivity and extended upper stopband is presented. It is comprised of two microstrip-to-microstrip vertical transitions with nonuniform slotline sections, which cascaded directly by a nonuniform microstrip connecting line. In terms of odd-/even-mode analysis, the in-band six poles which make up the wideband bandwidth can be explicitly derived. Moreover, the Chebyshev equal-ripple filtering responses with prescribed specifications can be conveniently achieved in desired passband by adopting synthesis theory. In order to suppress the second and third spurious harmonics, a modified circuit model based on synthesis method is further proposed. Two additional transmission zeros (TZs) TZ3, TZ4 can be generated at $3f_{0}$ and $5f_{0}$ respectively by adjusting impedance ratio of slotline stubs properly under the condition of $\theta _{\mathrm {s1}} = \theta /4$ and $\theta _{\mathrm {s2}} = \theta /2$ , which widen the upper stopband bandwidth greatly and enhance harmonic rejection level effectively. Meanwhile, combing with the inherent periodic finite-frequency TZ5, TZ6 and TZ7, the upper stopband could be extended to $6.22f_{0}$ with rejection level better than 14.6 dB. Besides, a pair of finite-frequency TZ1 and TZ2 caused by source-load coupling occur at both sides of passband to sharpen the cutoff slope of skirts remarkably. For verification, a prototype is fabricated and measured, the experimental results agree well with the theoretical predictions, which verify the validity of proposed method.

Abstract Background Protein lysine 2-hydroxyisobutyrylation (Khib) is a novel post-translational modification (PTM) discovered in cells or tissues of animals, microorganisms and plants in recent years. Proteome-wide identification of Khib-modified proteins has been performed in several plant species, suggesting that Khib-modified proteins are involved in a variety of biological processes and metabolic pathways. However, the protein Khib modification in soybean, a globally important legume crop that provides the rich source of plant protein and oil, remains unclear. Results In this study, the Khib-modified proteins in soybean leaves were identified for the first time using affinity enrichment and high-resolution mass spectrometry-based proteomic techniques, and a systematic bioinformatics analysis of these Khib-modified proteins was performed. Our results showed that a total of 4251 Khib sites in 1532 proteins were identified as overlapping in three replicates (the raw mass spectrometry data are available via ProteomeXchange with the identifier of PXD03650). These Khib-modified proteins are involved in a wide range of cellular processes, particularly enriched in biosynthesis, central carbon metabolism and photosynthesis, and are widely distributed in subcellular locations, mainly in chloroplasts, cytoplasm and nucleus. In addition, a total of 12 sequence motifs were extracted from all identified Khib peptides, and a basic amino acid residue (K), an acidic amino acid residue (E) and three aliphatic amino acid residues with small side chains (G/A/V) were found to be more preferred around the Khib site. Furthermore, 16 highly-connected clusters of Khib proteins were retrieved from the global PPI network, which suggest that Khib modifications tend to occur in proteins associated with specific functional clusters. Conclusions These findings suggest that Khib modification is an abundant and conserved PTM in soybean and that this modification may play an important role in regulating physiological processes in soybean leaves. The Khib proteomic data obtained in this study will help to further elucidate the regulatory mechanisms of Khib modification in soybean in the future.

In recent years, the government has actively set up computer programming courses to train those with the relevant talent; however, the learning performance of the students is not ideal. Therefore, in order to learn programming skills, students usually adopt note-taking strategies because, due to the pressure of the course, the teachers do not have sufficient time to help the students to fully understand the course content. This means that some students take notes without thinking, so their academic performance is usually poor. This study, therefore, proposes an innovative curriculum design that is based on the "Note-taking System and Teaching Strategy'' (NSTS), which combines learning style concepts and peer learning concepts to achieve student interaction and promote their thinking skills. In the learning activity, students are asked to search for additional supplementary material and to write their own notes, and then the members of group can read their notes and learn from them. However, the results of the study show that the NSTS curriculum design exhibits significant differences for improving the students' academic performance, and that it also has a certain influence on their learning motivation.

Objective To quantitatively measure the morphological parameters of femoral condyle based on MRI images so as to determine the effect of anterior-posterior femoral condyle geometry on patellar instability (PI) in patients with high-grade femoral trochlear dysplasia. Methods A total of 82 series of knee joint MRI images from 74 patients with high-grade femoral trochlear dysplasia who underwent MRI scanning in our hospital from January 2014 to January 2021 were retrospectively selected. According to the results of clinical diagnosis and radiological imaging, they were further divided into patellar dislocation group (n=45) and non-patellar dislocation group (n=37). Subsequently, the morphological parameters related to the femoral condyle of knee joint were measured quantitatively in the MRI images, including posterior condylar angle (PCA), width of medial-lateral epicondyle (MLW), medial posterior condyle width (MPCW), lateral posterior condyle width (LPCW), medial anterior condyle length (MACL), lateral anterior condyle length (LACL), medial posterior condyle length (MPCL), and lateral posterior condyle length (LPCL). Intraclass correlation coefficient (ICC) was used to assess the intra- and inter-observer reliability of the quantitative MRI measurements. Results The MPCW and LACL values were significantly lower in the patellar dislocation group than the non-patellar dislocation group (P < 0.05), whereas there were no statistical differences in PCA, MLW, LPCW, MACL, MPCL and LPCL values between the 2 groups. The measurement results of all parameters had good consistency within and among the observers (ICC≥0.80). Conclusion The patients of patellar dislocation have shorter lateral anterior femoral condyle and narrower medial posterior femoral condyle, which may be risk factors for dislocation in those with high-grade trochlear dysplasia.

ObjectiveThis study aimed to investigate the reliability of a deep neural network (DNN) model trained only on contrast-enhanced T1 (T1CE) images for predicting intraoperative cerebrospinal fluid (ioCSF) leaks in endoscopic transsphenoidal surgery (EETS).Methods396 pituitary adenoma (PA) cases were reviewed, only primary PAs with Hardy suprasellar Stages A, B, and C were included in this study. The T1CE images of these patients were collected, and sagittal and coronal T1CE slices were selected for training the DNN model. The model performance was evaluated and tested, and its interpretability was explored.ResultsA total of 102 PA cases were enrolled in this study, 51 from the ioCSF leakage group, and 51 from the non-ioCSF leakage group. 306 sagittal and 306 coronal T1CE slices were collected as the original dataset, and data augmentation was applied before model training and testing. In the test dataset, the DNN model provided a single-slice prediction accuracy of 97.29%, a sensitivity of 98.25%, and a specificity of 96.35%. In clinical test, the accuracy of the DNN model in predicting ioCSF leaks in patients reached 84.6%. The feature maps of the model were visualized and the regions of interest for prediction were the tumor roof and suprasellar region.ConclusionIn this study, the DNN model could predict ioCSF leaks based on preoperative T1CE images, especially in PAs in Hardy Stages A, B, and C. The region of interest in the model prediction-making process is similar to that of humans. DNN models trained with preoperative MRI images may provide a novel tool for predicting ioCSF leak risk for PA patients.

Pink bollworms severely affect the production of cotton. The method currently used for pink bollworm control is the planting of Bt (Bacillus thuringiensis) protein-expressing transgenic cotton. However, pink bollworms can develop strong resistance to Bt proteins in transgenic cotton because of the large planting area and long planting time of this crop, which severely affects the control of pink bollworms. Intestinal microorganisms play very important roles in insect growth, development and Bt resistance. However, the effect of intestinal microorganisms on pink bollworm Bt resistance is still unclear. The current study aimed to analyze the effect of intestinal microorganisms on the Bt resistance of pink bollworms. Intestinal microorganisms associated with Bt resistance were initially screened through Illumina MiSeq sequencing and analysis. The results showed that feeding with a mixture of gentamicin, Cry1Ac and an artificial diet could significantly increase the mortality of pink bollworm larvae compared with feeding with of a mixture of Cry1Ac and an artificial diet or an artificial diet alone. The microbial diversity, community structure and composition of the pink bollworm larval intestine were significantly influenced by feeding with a mixture of gentamicin, Cry1Ac and an artificial diet. Several intestinal bacteria with significantly altered abundances after treatment with gentamicin were preliminarily screened as potential resources for addressing Bt toxicity. This study provides useful strategies for addressing the Bt resistance of pink bollworms.