A Novel Ubiquitin Ligase Adaptor PTPRN Suppresses Seizure Susceptibility through Endocytosis of NaV1.2 Sodium Channels.

Intrinsic plasticity, a fundamental process enabling neurons to modify their intrinsic properties, plays a crucial role in shaping neuronal input-output function and is implicated in various neurological and psychiatric disorders. Despite its importance, the underlying molecular mechanisms of intrinsic plasticity remain poorly understood. In this study, a new ubiquitin ligase adaptor, protein tyrosine phosphatase receptor type N (PTPRN), is identified as a regulator of intrinsic neuronal excitability in the context of temporal lobe epilepsy. PTPRN recruits the NEDD4 Like E3 Ubiquitin Protein Ligase (NEDD4L) to NaV1.2 sodium channels, facilitating NEDD4L-mediated ubiquitination, and endocytosis of NaV1.2. Knockout of PTPRN in hippocampal granule cells leads to augmented NaV1.2-mediated sodium currents and higher intrinsic excitability, resulting in increased seizure susceptibility in transgenic mice. Conversely, adeno-associated virus-mediated delivery of PTPRN in the dentate gyrus region decreases intrinsic excitability and reduces seizure susceptibility. Moreover, the present findings indicate that PTPRN exerts a selective modulation effect on voltage-gated sodium channels. Collectively, PTPRN plays a significant role in regulating intrinsic excitability and seizure susceptibility, suggesting a potential strategy for precise modulation of NaV1.2 channels' function.

The Effects of Programmed Cell Death of Mesenchymal Stem Cells on the Development of Liver Fibrosis.

Mesenchymal stem cells have shown noticeable potential for unlimited self-renewal. They can differentiate into specific somatic cells, integrate into target tissues via cell-cell contact, paracrine effects, exosomes, and other processes and then regulate the target cells and tissues. Studies have demonstrated that transplantation of MSCs could decrease the expression and concentration of collagen in the liver, thereby reducing liver fibrosis. A growing body of evidence indicates that apoptotic MSCs could inhibit harmful immune responses and reduce inflammatory responses more effectively than viable MSCs. Accumulating evidence suggests that mitochondrial transfer from MSCs is a novel strategy for the regeneration of various damaged cells via the rescue of their respiratory activities. This study is aimed at reviewing the functions of MSCs and the related roles of the programmed cell death of MSCs, including autophagy, apoptosis, pyroptosis, and ferroptosis, as well as the regulatory pathogenic mechanisms of MSCs in liver fibrosis. Research has demonstrated that the miR-200B-3p gene is differentially expressed gene between LF and normal liver samples, and that the miR-200B-3p gene expression is positively correlated with the degree of liver fibrosis, suggesting that MSCs could inhibit liver fibrosis through pyroptosis. It was confirmed that circulating monocytes could deliver MSC-derived immunomodulatory molecules to different sites by phagocytosis of apoptotic MSCs, thereby achieving systemic immunosuppression. Accordingly, it was suggested that characterization of the programmed cell death-mediated immunomodulatory signaling pathways in MSCs should be a focus of research.

Diagnostic value and cost-effectiveness of FNA-CT versus FNAC for medullary thyroid carcinoma.

OBJECTIVE: To evaluate the diagnostic performance and cost-effectiveness of calcitonin assays in fine-needle aspiration washout fluid (FNA-CT) compared to fine-needle aspiration cytology (FNAC) for medullary thyroid carcinoma (MTC). METHODS: A total of 27,404 patients from three medical centres between January 2020 and May 2022 were screened for serum calcitonin (sCT). Of whom, 223 patients met endpoints and were enroled for analyses. Based on sCT levels, patients were divided into two groups (group 1: 10 pg/ml< sCT ≤100 pg/ml and group 2: sCT > 100 pg/ml). The diagnostic performance and cost-effectiveness of FNA-CT and FNAC were compared. RESULTS: Most patients (N = 25,228; 92.1%) with thyroid nodules had normal sCT levels. In group 1, 24 and 167 nodules were diagnosed as MTC and non-MTC lesions, respectively. FNA-CT showed better performance in diagnosing MTC than FNAC in terms of sensitivity (100.0% vs. 58.3%), negative predictive value (100.0% vs. 94.3%), and overall accuracy (100.0% vs. 94.7%). In group 2, 67 and 7 nodules were diagnosed as MTC and non-MTC lesions, respectively. The diagnostic performance of FNA-CT was superior to FNAC in terms of sensitivity (100.0% vs. 64.2%), negative predictive value (100.0% vs. 22.6%), and overall accuracy (100.0% vs. 67.6%). Furthermore, analysis from the decision tree model showed that FNA-CT was a cost-effective tool for diagnosing MTC lesions. CONCLUSIONS: FNA-CT can serve as an auxiliary and cost-effective approach for patients with indeterminate sCT levels to detect occult MTC lesions. FNA-CT can be recommended for patients with sCT >100 pg/ml to overcome the high false-negative rate of FNAC.

Willingness of Older Adults with Chronic Diseases to Receive a Booster Dose of Inactivated Coronavirus Disease 2019 Vaccine: A Cross-Sectional Study in Taizhou, China.

Vaccination is an important measure to control the spread of COVID-19 among elderly high-risk groups; however, the propensity to receive COVID-19 vaccine boosters has not been evaluated in these populations. Here, we aimed to investigate the willingness to receive a COVID-19 vaccine booster among the elderly chronic disease population in Taizhou, China. A cross-sectional, hospital-based survey was conducted in the outpatient department of a tertiary care hospital between 6 July and 11 August 2021 in Taizhou, China, and the data were uploaded to Wen-Juan-Xing, one of the largest online platforms used to collect survey data in China. The targeted population was non-oncology chronic disease patients aged 60 years and above. The minimum sample size was 229, determined by the G*Power software (v3.1.9.2, Heinrich-Heine-Universität Düsseldorf, Düsseldorf, Germany). A total of 254 patients with valid data were enrolled in this study, with a response rate of 82.5% (254/308). Chi-square tests and one-way binary regression were used to compare the proportions and the degree of influence of categorical factors. The magnitude of the effect for the comparisons was measured by Gramer's V. A multivariate binary logistic regression model was used to correct for confounders and to identify factors. All data were analyzed using SPSS v24.0 (IBM Corporation, Armonk, NY, USA). A total of 198 respondents (77.9%) were willing to receive a COVID-19 vaccine booster dose, and 77.6% of respondents were willing to receive the primary dose. Age < 70 years (OR 2.82), stable disease control (OR 2.79), confidence in the effectiveness of the COVID-19 vaccine (OR 3.11), and vaccine recipient (OR 5.02) were significantly associated with the willingness to receive a COVID-19 vaccine booster dose. Promoting primary dose vaccination is essential for advancing booster vaccination, and it is important to focus on elderly patients' confidence in the vaccine, in addition to strengthening health management and promoting disease stability. Follow-up studies should focus on elderly patients who belong to specific disease groups.

Adverse reactions to inactivated COVID-19 vaccination in patients with chronic liver disease: The effect of anxiety.

Studies have shown that patients with chronic liver disease are at a higher risk of contracting novel coronavirus pneumonia than healthy individuals, and many guidelines state that patients with chronic liver disease should be prioritized for COVID-19 vaccination, but there are a few studies on its safety in CLD patients. We aimed to evaluate the safety of the inactivated COVID-19 vaccine in patients with chronic liver disease, and the effect of anxiety on adverse reactions. A questionnaire survey for self-administered post-vaccination adverse reaction monitoring was conducted from June 17, 2021, to August 11, 2021, in patients with chronic liver disease attending a tertiary care hospital in Taizhou, China. We analyzed the data from of a total of 160 participants who scanned the QR code on social media to respond to the questionnaire. The overall incidence of adverse reactions after COVID-19 vaccination in patients with chronic liver disease was 44.4% (71/160), and the most common adverse reaction was local injection site reaction, accounting for 80.3% of adverse reactions (57/71). No serious adverse reactions were reported. Approximately 53.1% of the patients had anxiety about vaccination, and 51.8% of those who felt anxious reported adverse reactions. The safety of COVID-19 vaccination in patients with chronic liver disease is good, and there is a strong association between adverse reactions and vaccine anxiety. Pre-vaccination education for patients with vaccine anxiety and psychological counseling may reduce reports of adverse reactions and improve patients' confidence in the vaccine.

Relationship between knowledge, attitudes, and practices and COVID-19 vaccine hesitancy: A cross-sectional study in Taizhou, China.

Objective: This study aimed to explore COVID-19 vaccine hesitancy in Chinese adults and analyzed the relationship between knowledge, attitudes, practices (KAP), and COVID-19 vaccine hesitancy. Methods: A population-based self-administered online survey was conducted in Taizhou, China to evaluate the population's hesitancy to receive COVID-19 vaccination. A total of 2.463 adults received the invitation for the survey through WeChat (A Chinese app that is used for chat, social media, and mobile payment), and 1.788 interviewees answered the structured questionnaire. The overall response rate was 72.6%. Results: Total 45.2% of people were hesitant about the COVID-19 vaccination. Using binary logistic regression analysis, we found low perception of safety (Model 3: Odds ratio = 2.977, Confidence interval: 2.237-3.963) and efficacy (Model 3: OR = 1.904, 95%CI: 1.462-2.479) of the COVID-19 vaccine in adults is the most important risk factor for COVID-19 vaccine hesitation. People who know more about COVID-19 vaccination are less hesitant (Model 2: OR = 0.967, 95% CI: 0.951-0.983). People who did not seek information independently about the COVID-19 vaccine are more likely to be skeptical (Model 4: OR = 1.300, 95% CI: 1.058-1.598, P = 0.013). Conclusion: In China, the population had higher levels of COVID-19 vaccine hesitation, and their knowledge of the COVID-19 vaccine, perceptions of safety and efficacy, and physical health status were significantly associated with vaccine hesitation. These results provide ideas for promoting COVID-19 vaccination and intervention and have far-reaching implications for further strengthening research on vaccine hesitancy in COVID-19 and exploring strategies for COVID-19 vaccine promotion.

Adaptive Cross Entropy for ultrasmall object detection in Computed Tomography with noisy labels.

Conventional size object detection has been extensively studied, whereas researches concerning ultrasmall object detection are rare due to lack of dataset. Here, considering that the stapes in the ear is the smallest bone in our body, we have collected the largest stapedial otosclerosis detection dataset from 633 stapedial otosclerosis patients and 269 normal cases to promote this direction. Nevertheless, noisy classification labels in our dataset are inevitable due to various subjective and objective factors, and this situation prevails in various fields. In this paper, we propose a novel and general noise tolerant loss function named Adaptive Cross Entropy (ACE) which needs no fine-tuning of hyperparameters for training with noisy labels. We provide both theoretical and empirical analyses for the proposed ACE loss and demonstrate its effectiveness in multiple public datasets. Besides, we find high-resolution representations crucial for ultrasmall object detection and present an auxiliary backbone called W-Net to address it accordingly. Extensive experiments demonstrate that the proposed ACE loss is able to boost the diagnosis performance under noisy label setting by a large margin. Furthermore, our W-Net can help extract sufficient high-resolution representations specialized for ultrasmall objects and achieve even better results. Hopefully, our work could provide more clues for future research on ultrasmall object detection and learning with noisy labels.

The impact of the Omicron epidemic on the health behavior in Cape Town, South Africa.

Background: South Africa was the first country with a case of Omicron variant infection diagnosed; therefore, this study aimed to elucidate the impact of the Omicron mutant strain outbreak on the health behavior of the South African population and encourage the population to adopt timely protective behaviors against Omicron mutant strain infection. Study design and methods: This was a population-based, cross-sectional study conducted in Cape Town, South Africa, in December 2021. We distributed 300 questionnaires to adults aged >18 years, and they were all returned. Results: Of the South African population, 60.3% expressed a high level of concern regarding Omicron; 89.3% improved on at least one of the following three health behaviors: mask-wearing, washing hands, and reducing socialization; and only 10.7% exhibited no improvement in health behaviors. Of these, 71.3% and 57.0% increased the length of time they wore a mask and washed their hands, respectively, and 47% decreased the number of times they socialized. Age, residence, education level, chronic disease, and whether they had received the COVID-19 vaccine were significantly different (p < 0.05) between the presence and absence of enhanced health behaviors. The levels of concern and knowledge regarding the Omicron virus significantly influenced health-behavior change (all P < 0.05). Conclusion: There has been a positive change in the South African population toward adopting mask-wearing, hand washing, and reducing socialization in response to the Omicron virus strain epidemic. Based on one health approach, it is important to focus on populations with chronic diseases, those who have not yet received the COVID-19 vaccine, and other populations with low rates of health behavior change.

Combination of CTLA-4 blockade with MUC1 mRNA nanovaccine induces enhanced anti-tumor CTL activity by modulating tumor microenvironment of triple negative breast cancer.

The immunosuppressive tumor microenvironment (TME) is the main reason for the failure of many immunotherapies that directly stimulate anti-tumor immune response. Anti-CTLA-4 antibody may reduce effector regulatory T (Treg) cell numbers and their suppressive activity in the TME. We have previously reported that combination of anti-CTLA-4 antibody with MUC1 mRNA nanovaccine may mutually enhance each single treatment. But the enhancement mechanism of therapeutic efficacy of MUC1 mRNA nanovaccine plus anti-CTLA-4 monoclonal antibody (mAb) is unknown. In this study, anti-tumor CTL activity induced by combination of CTLA-4 Blockade with MUC1 mRNA nanovaccine and immunosuppressive factors in the TME of triple negative breast cancer were investigated. The results demonstrated that combined therapy with nanovaccine and anti-CTLA-4 mAb could induce stronger anti-tumor CTL response than each monotherapy, result in significantly decreased numbers of myeloid-derived suppressor cells (MDSC), Treg cells, tumor-associated fibroblasts (TAFs) and tumor vasculature in the TME, downregulated levels of interleukin-6, tumor necrosis factor-α and transforming growth factor-ß, and significantly upregulated levels of IFN-γ and interleukin-12 as well as increased number of CD8+ T cell, and appear more effective than either nanovaccine or anti-CTLA-4 mAb alone at increasing level of apoptosis in tumor cells. In addition, combination immunotherapy could significantly downregulated the signal transducer and activator of transcription 3 (STAT3) signal pathway. Therefore, it can be concluded that combination of CTLA-4 blockade with MUC1 mRNA nanovaccine enhances anti-tumor cytotoxic T-lymphocyte activity by reducing immunosuppressive TME and inhibiting tumor-promoting STAT3 signaling pathway.

The use of explainable artificial intelligence to explore types of fenestral otosclerosis misdiagnosed when using temporal bone high-resolution computed tomography.

BACKGROUND: The purpose of this study was to explore the common characteristics of fenestral otosclerosis (OS) which are misdiagnosed, and develop a deep learning model for the diagnosis of fenestral OS based on temporal bone high-resolution computed tomography scans. METHODS: We conducted a study to explicitly analyze the clinical performance of otolaryngologists in diagnosing fenestral OS and developed an explainable deep learning model using 134,574 temporal bone high-resolution computed tomography (HRCT) slices collected from 1,294 patients for the automatic diagnosis of fenestral OS. We prospectively created an external test set with 31,774 CT slices from 144 patients, which contained 86 fenestral OS ears and 202 normal ears and used it to evaluate the performance of our otosclerosis-Logical Neural Network (LNN) model to assess its potential clinical utility. In addition, we compared the diagnostic acumen of seven otolaryngologists with the otosclerosis-LNN approach in the clinical test set, which was mixed with 78 fenestral OS and 62 normal ears. Finally, to evaluate the assisting value of the model, the seven participants were again invited to classify all cases in the clinical test set after referring to the diagnostic results of the model, to which they were blinded. RESULTS: The diagnostic performance of otologists was not satisfactory, and those CT samples which were misdiagnosed had similar characteristics. Based on this finding, we defined three subtypes of fenestral OS lesions that are suitable for clinical diagnosis guidance: "focal", "transitional", and "typical" fenestral OS. The most encouraging result is that the model achieved an area under the curve (AUC) of 99.5% (per-ear-sensitivity of 96.4%, per-ear-specificity of 98.9%) on the prospective unknown external test. Furthermore, we used this model to assist otologists and observed a consistent and significant improvement in diagnostic performance, especially for the newly defined focal and transitional fenestral OS, which led to the initial high misdiagnosis rate. CONCLUSIONS: Our findings of the fine-grained classification of fenestral OS could have implications for future diagnosis and prevention programs. In addition, our deep OS localization network is an effective approach providing assistance to otologists to deal with the significant challenge of the misdiagnosis of fenestral OS.

[Preparation of in vitro transcribed mouse MUC1 mRNA and its expression in NIH/3T3 cells].

Objective To obtain in vitro transcription of mouse mucin 1(MUC1) mRNA and express it in NIH/3T3 cells. Methods The MUC1 gene was amplified from mouse 4T1 cells by reverse transcription PCR (RT-PCR), and then inserted into the eukaryotic expression vector pcDNA3.1(+) to construct the recombinant vector pcDNA3.1(+)-MUC1. After identification by restriction enzyme cutting, the recombinant plasmid was used as a PCR template for the amplification of the MUC1-HA TAG fusion gene fragment with the reverse primer containing hemagglutinin tag (HA TAG) sequence. Subsequently, the MUC1-HA TAG fusion gene was cloned into the expression vector pcDNA3.1(+). After double restriction enzyme digestion and DNA sequence identification, the recombinant plasmid encoding MUC1-HA TAG was used as a PCR template for the amplification of a PCR product containing T7 promoter and the MUC1-HA TAG fusion gene fragment as an in vitro transcription template to obtain the modified MUC1-HA TAG mRNA by in vitro transcription, tailing and purification. The resulting MUC1-HA TAG mRNA was transfected into NIH/3T3 cells by various transfection reagents and the expression of the fusion protein MUC1-HA TAG was detected by Western blot analysis. Results The MUC1 gene amplified by RT-PCR was about 1 900 bp in length. The restriction enzyme digestion and sequence analysis confirmed that the MUC1-HA TAG fusion gene had been successfully inserted into pcDNA3.1(+), and the insertion sequence was identical to the MUC1 gene sequence of the mouse in GenBank database. HA TAG was correctly inserted in the downstream of MUC1 gene and the reading frame was correct. Western blot analysis indicated that the in vitro transcriptionally modified MUC1-HA TAG mRNA could be expressed in NIH/3T3 cells. Conclusion In vitro transcriptionally modified mouse MUC1-HA TAG mRNA can be translated and expressed in mammalian NIH/3T3 cells.