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The regional distribution of neurofibrillary tangles of hyperphosphorylated tau aggregates is associated with the progression of Alzheimer’s disease (AD). Misfolded proteopathic tau recruits naïve tau and templates its misfolding and aggregation in a prion-like fashion, which is believed to be the molecular basis of propagation of tau pathology. A practical way to assess tau seeding activity is to measure its ability to recruit/bind other tau molecules and to induce tau aggregation. Based on the properties of proteopathic tau, here we report the development of two simple assays to assess tau seeding activity ----- capture assay in vitro and seeded-tau aggregation assay in cultured cells. In the capture assay, proteopathic tau was applied onto a nitrocellulose membrane and the membrane was incubated with cell lysate containing HA-tagged tau151-391 (HA-tau151-391). The captured tau on the membrane was determined by immuno-blots developed with anti-HA. For the seeded-tau aggregation assay, HEK-293FT cells transiently expressing HA-tau151-391 were treated with proteopathic tau in the presence of Lipofectamine 2000 and then lysed with RIPA buffer. RIPA-insoluble fraction containing aggregated tau was obtained by ultracentrifugation and analyzed by immuno-blot developed with anti-HA. To validate these two assays, we assessed the seeding activity of tau in the middle frontal gyrus, middle temporal gyrus and basal forebrain of AD and control brains and found that AD, but not control, brain extracts effectively captured and seeded tau151-391 aggregation. Basal forebrain contained less phospho-tau and tau seeding activity. The levels of captured tau or seeded-tau aggregates were positively correlated to the levels of phospho-tau, Braak stages and tangle sores. These two assays are specific and sensitive and can be carried out in a regular biomedical laboratory setting by using routine biochemical techniques.

IntroductionTo explore the distribution of Isthmin-1 (ISM1) level and its association with isolated post-challenge hyperglycemia (IPH).MethodsA total of 522 participants without a history of diabetes were invited to attend a standard 75g 2-h oral glucose tolerance test (OGTT), and 71 subjects were further invited for a 3-h oral minimal model test. Insulin sensitivity and β-cell function were evaluated using both HOMA and estimated from OGTT. Circulating ISM1 levels were determined by a commercially available ELISA kit.ResultsA total of 76 (14.6%) participants were diagnosed as IPH, accounting for 61.3% of the newly diagnosed diabetes. ISM1 levels were significantly higher in men than in women (1.74 ng/mL versus 0.88 ng/mL). The inverse correlation between ISM1 and β-cell function and IPH was only significant in men. After multivariate adjustment, per unit increment in ISM1 was associated with 0.68-fold (95% CI: 0.49-0.90) reduced odds ratio (OR) of IPH in men. Compared to men with the lowest ISM1 levels, the adjusted OR of IPH with the highest ISM1 levels decreased by 73% (95% CI: 0.11-0.61). Moreover, incorporation of ISM1 into the New Chinese Diabetes Risk Score (NCDRS) model yielded a substantial improvement in net reclassification improvement of 58% (95% CI: 27%-89%) and integrated discrimination improvement of 6.4% (95% CI: 2.7%-10.2%) for IPH.ConclusionsISM1 was significantly and independently associated with IPH, and serves as a feasible biomarker for the early identification of men with high risk of IPH.

Background:In recent years, diabetic kidney disease (DKD) has emerged as a prominent factor contributing to end-stage renal disease. Tubulointerstitial inflammation and lipid accumulation have been identified as key factors in the development of DKD. Earlier research indicated that Astragaloside IV (AS-IV) reduces inflammation and oxidative stress, controls lipid accumulation, and provides protection to the kidneys. Nevertheless, the mechanisms responsible for its protective effects against DKD have not yet been completely elucidated.Purpose:The primary objective of this research was to examine the protective properties of AS-IV against DKD and investigate the underlying mechanism, which involves CD36, reactive oxygen species (ROS), NLR family pyrin domain containing 3 (NLRP3), and interleukin-1β (IL-1β).Methods:The DKD rat model was created by administering streptozotocin along with a high-fat diet. Subsequently, the DKD rats and palmitic acid (PA)-induced HK-2 cells were treated with AS-IV. Atorvastatin was used as the positive control. To assess the therapeutic effects of AS-IV on DKD, various tests including blood sugar levels, the lipid profile, renal function, and histopathological examinations were conducted. The levels of CD36, ROS, NLRP3, Caspase-1, and IL-1β were detected using western blot analysis, PCR, and flow cytometry. Furthermore, adenovirus-mediated CD36 overexpression was applied to explore the underlying mechanisms through in vitro experiments.Results:In vivo experiments demonstrated that AS-IV significantly reduced hyperglycemia, dyslipidemia, urinary albumin excretion, and serum creatinine levels in DKD rats. Additionally, it improved renal structural abnormalities and suppressed the expression of CD36, NLRP3, IL-1β, TNF-α, and MCP-1. In vitro experiments showed that AS-IV decreased CD36 expression, lipid accumulation, and lipid ROS production while inhibiting NLRP3 activation and IL-1β secretion in PA-induced HK-2 cells.Conclusion:AS-IV alleviated renal tubule interstitial inflammation and tubule epithelial cell apoptosis in DKD rats by inhibiting CD36-mediated lipid accumulation and NLRP3 inflammasome activation.

IntroductionTrophoblast cell surface antigen 2 (TROP2; EpCAM2) is a transmembrane glycoprotein which is closely related to EpCAM (EpCAM; EpCAM1). Both proteins share partial overlapping functions in epithelial development and EpCAM expression but have not been comparatively analyzed together in bladder carcinomas. TROP2 constitutes the target for the antibody-drug conjugate Sacituzumab govitecan (SG; TrodelvyTM) which has been approved for treatment of metastatic urothelial carcinoma by the United States Food and Drug administration (FDA) irrespective of its TROP2 expression status. MethodsTo evaluate the potential clinical significance of subtle differences in TROP2 and EpCAM expression in urothelial bladder cancer, both proteins were analyzed by multiplex fluorescence immunohistochemistry in combination with a deep-learning based algorithm for automated cell detection on more than 2,700 urothelial bladder carcinomas in a tissue microarray (TMA) format. ResultsThe staining pattern of TROP2 and EpCAM were highly similar. For both proteins, the staining intensity gradually decreased from pTa G2 low grade (TROP2: 68.8±36.1; EpCAM: 21.5±11.7) to pTa G2 high grade (64.6±38.0; 19.3±12.2) and pTa G3 (52.1±38.7; 16.0±13.0, p

Rock image classification is a significant part of geological research. Compared with traditional image classification methods, rock image classification methods based on deep learning models have the great advantage in terms of automatic image features extraction. However, the rock classification accuracies of existing deep learning models are unsatisfied due to the weak feature extraction ability of the network model. In this study, a deep residual neural network (ResNet) model with the transfer learning method is proposed to establish the corresponding rock automatic classification model for seven kinds of rock images. ResNet34 introduces the residual structure to make it have an excellent effect in the field of image classification, which extracts high-quality rock image features and avoids information loss. The transfer learning method abstracts the deep features from the shallow features, and better express the rock texture features for classification in the case of fewer rock images. To improve the generalization of the model, a total of 3,82,536 rock images were generated for training via image slicing and data augmentation. The network parameters trained on the Texture Library dataset which contains 47 types of texture images and reflect the characteristics of rocks are used for transfer learning. This pre-trained weight is loaded when training the ResNet34 model with the rock dataset. Then the model parameters are fine-tuned to transfer the model to the rock classification problem. The experimental results show that the accuracy of the model without transfer learning reached 88.1%, while the model using transfer learning achieved an accuracy of 99.1%. Aiming at geological engineering field investigation, this paper studies the embedded deployment application of the rock classification network. The proposed rock classification network model is transplanted to an embedded platform. By designing a rock classification system, the off-line rock classification is realized, which provides a new solution for the rock classification problem in the geological survey. The deep residual neural network and transfer learning method used in this paper can automatically classify rock features without manually extracting. These methods reduce the influence of subjective factors and make the rock classification process more automatic and intelligent.

PurposeTo systematically review the clinical value of three imaging examinations (Magnetic Resonance Imaging, Computed Tomography, and myelography) in the diagnosis of Lumbar Disc Herniation.MethodsDatabases including PubMed, Embase, The Cochrane Library, Web of Science, CBM, CNKI, WanFang Data, and VIP were electronically searched to collect relevant studies on three imaging examinations in the diagnosis of Lumbar Disc Herniation from inception to July 1, 2021. Two reviewers using the Quality Assessment of Diagnostic Accuracy Studies-2 tool independently screened the literature, extracted the data, and assessed the risk of bias of included studies. Then, meta-analysis was performed by using Meta-DiSc 1.4 software and Stata 15.0 software.ResultsA total of 38 studies from 19 articles were included, involving 1,875 patients. The results showed that the pooled Sensitivity, pooled Specificity, pooled Positive Likelihood Ratio, pooled Negative Likelihood Ratio, pooled Diagnostic Odds Ratio, Area Under the Curve of Summary Receiver Operating Characteristic, and Q* were 0.89 (95%CI: 0.87–0.91), 0.83 (95%CI: 0.78–0.87), 4.57 (95%CI: 2.95–7.08), 0.14 (95%CI: 0.09–0.22), 39.80 (95%CI: 18.35–86.32), 0.934, and 0.870, respectively, for Magnetic Resonance Imaging. The pooled Sensitivity, pooled Specificity, pooled Positive Likelihood Ratio, pooled Negative Likelihood Ratio, pooled Diagnostic Odds Ratio, Area Under the Curve of Summary Receiver Operating Characteristic, and Q* were 0.82 (95%CI: 0.79–0.85), 0.78 (95%CI: 0.73–0.82), 3.54 (95%CI: 2.86–4.39), 0.19 (95%CI: 0.12–0.30), 20.47 (95%CI: 10.31–40.65), 0.835, and 0.792, respectively, for Computed Tomography. The pooled Sensitivity, pooled Specificity, pooled Positive Likelihood Ratio, pooled Negative Likelihood Ratio, pooled Diagnostic Odds Ratio, Area Under the Curve of Summary Receiver Operating Characteristic, and Q* were 0.79 (95%CI: 0.75–0.82), 0.75 (95%CI: 0.70–0.80), 2.94 (95%CI: 2.43–3.56), 0.29 (95%CI: 0.21–0.42), 9.59 (95%CI: 7.05–13.04), 0.834, and 0.767 respectively, for myelography.ConclusionThree imaging examinations had high diagnostic value. In addition, compared with myelography, Magnetic Resonance Imaging had a higher diagnostic value.

Several studies have demonstrated that the T-cell receptor (TCR) repertoire is associated with prognosis and immune therapy response in several types of cancer. However, the comprehensive features of TCR repertoire in tumor-infiltrating and circulating T cells, as well as its clinical significance of diagnosis in hepatocellular carcinoma (HCC) patients, are still unknown. In this study, we perform paired tumor/peritumoral tissues and peripheral blood samples from 58 patients with HCC and sequenced them with high-throughput TCR to comprehensively analyze the characteristics of TCR and the clinical significance of peripheral TCR sequence. By exploring the abundance and diversity of TCR repertoires, we observe that there was a significantly higher TCR diversity in peripheral blood than in tumoral and peritumoral tissues, while tumoral and peritumoral tissues showed similar TCR diversity. A substantial difference in the usage frequencies of several Vβ, Jβ genes, and TCRβ VJ pairings was found among three types of tissues. Moreover, we reveal that HCC patients have a unique profile of TCR repertoire in peripheral blood in contrast to healthy individuals. We further establish an HCC diagnostic model based on TCRβ VJ pairing usage in peripheral blood, which yields a best-fit area under the curve (AUC) of 0.9746 ± 0.0481 (sensitivity = 0.9675 ± 0.0603, specificity = 0.9998 ± 0.0007, average of 100 repeats) in the test set. Our study describes the characteristics of tissue infiltration and circulating T-cell bank in patients with HCC and shows the potential of using circulating TCR sequence as a biomarker for the non-invasive diagnosis of patients with HCC.

BackgroundGlobally, stomach adenocarcinoma (STAD)’s high morbidity and mortality should arouse our urgent attention. How long can STAD patients survive after surgery and whether novel immunotherapy is effective are questions that our clinicians cannot escape.MethodsVarious R packages, GSEA software, Metascape, STRING, Cytoscape, Venn diagram, TIMER2.0 website, TCGA, and GEO databases were used in our study.ResultsIn the TCGA and GEO, macrophage abundance of STAD tissues was significantly higher than that of adjacent tissues and was an independent prognostic factor, significantly related to the overall survival (OS) of STAD patients. Between the high- and low- macrophage abundance, we conducted differential expression, univariate and multivariate Cox analysis, and obtained 12 candidate genes, and finally constructed a 3-gene signature. Both low macrophage abundance group and group D had higher TMB and PD-L1 expression. Furthermore, top 5 common gene-mutated STAD tissues had lower macrophage abundance. Macrophage abundance and 3 key genes expression were also lower in the Epstein-Barr Virus (EBV) and HM-indel STAD subtypes and significantly correlated with the tumor microenvironment score. The functional enrichment and ssGSEA revealed 2 signatures were similar and closely related to BOQUEST_STEM_CELL_UP, including genes up-regulated in proliferative stromal stem cells. Hsa-miR-335-5p simultaneously regulated 3 key genes and significantly related to the expression of PD-L1, CD8A and PDCD1.Conclusionmacrophage abundance and 3-gene signature could simultaneously predict the OS and immunotherapy efficacy, and both 2 signatures had remarkable similarities. Hsa-miR-335-5p and BOQUEST_STEM_CELL_UP might be novel immunotherapy targets.