Objective To investigate the role of lncRNA SNHG1 in homocysteine-induced pyroptosis of podocyte.Methods Cbs+/-mice were randomly divided into two groups:a normal diet group(ND)and a high me-thionine diet group(HMD).Western blotting was used to detect the protein expression levels of Caspase-1,Cleaved Caspase-1,and NLRP3.Mouse renal glomerular podocytes were cultured in vitro,and then assigned into a control group(Control,0 μmol/L Hcy)and a homocysteine intervention group(Hcy,80 μmol/L Hcy).Western blotting was used to detect the protein expression levels of Caspase-1,Cleaved Caspase-1,and NLRP3.Mouse renal glomerular podocyion group(OE-NC + Hcy)and the lncSNHG1 overexpression + homocysteine intervention group(OE-SNHG1 + Hcy)were also established.After 48 hours of intervention,Real-time fluorescence quantita-tive PCR was used to detect the expression of lncSNHG1 in podocytes after Hcy intervention.Western blot was used to detect the expressions of Caspase-1,Cleaved Caspase-3 and NLRP3.Immunofluorescence was used to de-tect the expression levels of GSDMD and GSDMD-N.ELISA was used to detect the contents of IL-1β and IL-18.Results(1)In the animal experiments,the expression levels of pyroptosis-related proteins Caspase-1,Cleaved Caspase-1,NLRP3,GSDMD,and GSDMD-N were all increased in the HMD group compared with the ND group.(2)In the cellular experiments,the expression levels of Caspase-1,Cleaved Caspase-1,NLRP3,GSDMD,and GSDMD-N were all increased in the Hcy group compared with the Control group,and the contents of pyroptosis-mediated inflammatory factors IL-1β and IL-18 were increased as well.(3)In the cellular experiments,the expres-sion of lncSNHG1 was increased in the Hcy group compared with the control group.After transduction with lnc-SNHG1 lentivirus,the expression of lncSNHG1 was increased in the OE-SNHG1 group,compared with the control group and the OE-NC group.(4)In the cellular experiments,the expressions of pyroptosis-related proteins Cas-pase-1,Cleaved Caspase-1,NLRP3,GSDMD,and GSDMD-N were increased compared with the OE-NC+Hcy group,and the contents of pyroptosis-mediated inflammatory factors IL-1β and IL-18 were increased in the OE-SNHG1+Hcy group.Conclusion These results indicate that lncSNHG1 may play a role in promoting Hcy induced podocytepyroptosis.

Aldehyde oxidase (AOX) is a molybdoenzyme that is primarily expressed in the liver and is involved in the metabolism of drugs and other xenobiotics. AOX-mediated metabolism can result in unexpected outcomes, such as the production of toxic metabolites and high metabolic clearance, which can lead to the clinical failure of novel therapeutic agents. Computational models can assist medicinal chemists in rapidly evaluating the AOX metabolic risk of compounds during the early phases of drug discovery and provide valuable clues for manipulating AOX-mediated metabolism liability. In this study, we developed a novel graph neural network called AOMP for predicting AOX-mediated metabolism. AOMP integrated the tasks of metabolic substrate/non-substrate classification and metabolic site prediction, while utilizing transfer learning from ¹³C nuclear magnetic resonance data to enhance its performance on both tasks. AOMP significantly outperformed the benchmark methods in both cross-validation and external testing. Using AOMP, we systematically assessed the AOX-mediated metabolism of common fragments in kinase inhibitors and successfully identified four new scaffolds with AOX metabolism liability, which were validated through in vitro experiments. Furthermore, for the convenience of the community, we established the first online service for AOX metabolism prediction based on AOMP, which is freely available at https://aomp.alphama.com.cn.

Objective To explore the value of automated functional imaging(AFI)in predicting coronary artery stenosis in patients without ventricular wall motion abnormalities.Methods A total of 40 patients without ventricular wall motion abnormalities under two-dimensional echocardiography and confirmed coronary artery heart diseases(CHD)(coronary stenosis≥70％)by coronary angiography(CAG)at the Xinxiang Central Hospital from July 2018 to September 2019 were selected as the research subjects.The detection rates of coronary artery stenosis ≥70％by AFI and CAG were compared.With reference to CAG as the gold standard,the predictive value of AFI for coronary artery stenosis ≥70％was evaluated.Results There was no significant difference in the detection rates of coronary artery stenosis ≥70％by AFI and CAG(x2=1.667,P＞0.05).The predictive efficacy of AFI for coronary artery stenosis ≥70％was as follows:a sensitivity of 100％,a specificity of 63.6％,the positive predictive value of 69.2％,the negative predictive value of 100％,and an accuracy of 80％for predicting stenosis ≥70％in the left anterior descending artery;a sensitivity of 56.2％,a specificity of 91.6％,the positive predictive value of 81.8％,the negative predictive value of 75.8％,and an accuracy of 77.5％for predicting stenosis ≥70％in the left circumflex artery;a sensitivity of 95.6％,a specificity of 47.0％,the positive predictive value of 70.9％,the negative predictive value of 88.0％,and an accuracy of 75.0％for predicting stenosis ≥70％in the right coronary artery;the overall sensitivity of 85.9％,the overall specificity of 69.8％,the overall positive predictive value of 72.0％,the overall negative predictive value of 84.6％,and the overall accuracy of 77.5％.Conclusion AFI can provide a sensitive,objective,non-invasive,and inexpensive examination method for the early clinical forecast of coronary artery stenosis.

In response to the limitation of some medical image segmentation models for rectal cancer auxiliary diagnosis that are only applicable to single-modality images,a medical image segmentation method based on a cross attention mechanism that is applicable to both CT and MRI modalities is presented.Considering the different feature representations of CT and MRI images,a cross attention mechanism is proposed to unify the feature representations of the two types of images.In view of the small lesions on rectal cancer images,an improved Swin Transformer segmentation network with 3 branches is established,and the cross attention mechanism is incorporated into it,enabling the model to segment lesion areas in both types of images.The proposed method is validated using CT and MRI image data from patients with rectal cancer.Compared with ADDA,CycleGAN,and SIFA methods,the proposed method improves the accuracy by 2.94%,3.05%,0.71%on CT images,and 3.31%,4.55%,1.76%on MRI images,respectively,demonstrating its superior segmentation performance for both types of images.

Purpose: Prostate cancer (PCa) is an epithelial malignancy that originates in the prostate gland and is generally categorized into low, intermediate, and high-risk groups. The primary diagnostic indicator for PCa is the measurement of serum prostate-specific antigen (PSA) values. However, reliance on PSA levels can result in false positives, leading to unnecessary biopsies and an increased risk of invasive injuries. Therefore, it is imperative to develop an efficient and accurate method for PCa risk stratification. Many recent studies on PCa risk stratification based on clinical data have employed a binary classification, distinguishing between low to intermediate and high risk. In this paper, we propose a novel machine learning (ML) approach utilizing a stacking learning strategy for predicting the tripartite risk stratification of PCa. Methods: Clinical records, featuring attributes selected using the lasso method, were utilized with 5 ML classifiers. The outputs of these classifiers underwent transformation by various nonlinear transformers and were then concatenated with the lasso-selected features, resulting in a set of new features. A stacking learning strategy, integrating different ML classifiers, was developed based on these new features. Results: Our proposed approach demonstrated superior performance, achieving an accuracy of 0.83 and an area under the receiver operating characteristic curve value of 0.88 in a dataset comprising 197 PCa patients with 42 clinical characteristics. Conclusions This study aimed to improve clinicians’ ability to rapidly assess PCa risk stratification while reducing the burden on patients. This was achieved by using artificial intelligence-related technologies as an auxiliary method for diagnosing PCa.

ObjectiveTo investigate the influence of sarcopenia on bone mass loss， the risk factors for bone mass loss in liver cirrhosis， and the correlation between body composition and bone mineral density （BMD） by comparing the clinical features of bone mass loss in patients with liver cirrhosis. MethodsA total of 92 patients who were hospitalized and diagnosed with liver cirrhosis in Department of Gastroenterology， The Second Affiliated Hospital of Kunming Medical University， from April to December of 2022 were enrolled， and based on the results of dual-energy X-ray absorptiometry， they were divided into bone mass loss group （osteopenia/osteoporosis） with 57 patients and normal bone mass group with 35 patients. The two groups were compared in terms of general data， laboratory examination， imaging data， and body composition analysis. The independent samples t-test or the Mann-Whitney U test was used for comparison of continuous data between two groups， and the chi-square test or the continuity correction chi-square test was used for comparison of categorical data between two groups； Pearson correlation analysis and Spearman correlation analysis were used to investigate correlation； a binary logistic regression analysis was used to investigate the risk factors for bone mass loss in liver cirrhosis. ResultsCompared with the normal bone mass group， the bone mass loss group had significantly higher age （t=-3.597， P<0.05）， proportion of female patients （χ²=8.393， P<0.05）， N-terminal middle molecular fragment of osteocalcin （N-MID） （Z=-3.068， P<0.05）， β isomer of C-terminal telopeptide of type I collagen （β-CTX） （t=-2.784， P<0.05）， and proportion of patients with sarcopenia （χ²=13.884， P<0.05） and significantly lower calcitonin （CT） （Z=-2.340， P<0.05） and L3 skeletal muscle index （L3-SMI） （t=4.621， P<0.05）. Compared with the normal bone mass group， the bone mass loss group had significantly lower total muscle mass （Z=-2.952， P<0.05）， right upper limb muscle mass （Z=-2.929， P<0.05）， left upper limb muscle mass （Z=-2.680， P<0.05）， right lower limb muscle mass （Z=-3.366， P<0.05）， left lower limb muscle mass （Z=-3.374， P<0.05）， presumed bone mass （t=2.842， P<0.05）， body water mass （Z=-2.779， P<0.05）， basal metabolic rate （BMR） （Z=-3.153， P<0.05）， and BMD of L1— L4 and femoral neck （t=9.789， t=10.280， t=10.832， Z=-7.298， t=8.945， all P<0.05）. Total muscle mass， muscle mass of trunk and limbs， presumed bone mass， BMR， and body water mass in body component analysis were positively correlated with L1 — L4 BMD and femoral neck BMD （all P<0.05）， and fat mass was positively correlated with L1 — L4 BMD （all P<0.05）. Sarcopenia （odds ratio ［OR］=8.737， 95% confidence interval ［CI］： 2.237 — 34.129， P=0.002）， age （OR=1.094， 95%CI： 1.019 — 1.175， P=0.013）， and N-MID （OR=1.095， 95%CI： 1.019 — 1.176， P=0.014） were independent risk factors for bone mass loss in patients with liver cirrhosis. ConclusionOld age， female sex， sarcopenia， elevated N-MID， elevated β-CTX， reduction in CT， low muscle mass， low presumed bone mass， low BMR， and low body water mass are the features of bone mass loss in patients with liver cirrhosis， and sarcopenia， age， and N-MID are independent risk factors for bone mass loss in patients with liver cirrhosis. Detailed assessment of body composition changes can help to identify abnormal BMD in patients with liver cirrhosis.

Endoplasmic reticulum (ER) stress is involved in the development and progression of tumors.In recent years, great attention has been paid to the study of the interplay of ER stress and T cell differentiation and functionality.Intense ER stress in the tumor-infiltrating T cells exacerbates T cell exhaustion and impairs T cell anti-tumor immunity.Therefore, a variety of ER stress inhibitors have been developed and utilized to alleviate T cell exhaustion, which improves T cell function in tumor microenvironment.Furthermore, the downregulation of several circadian clock genes like Per1 and Per2 also aggravates T cell exhaustion, and the key downstream effector molecules in ER stress regulate the transcription of Per family, thus enhancing the T cell function.In the present manuscript, we particularly summarize how ER stress impacts the anti-tumor immunity of T cells, and further discuss potential strategies for improving tumor immunotherapy via targeting ER stress.

Chemotherapeutic drugs are the mainstay of treatment on mid-advanced tumors. In recent years, more and more studies have shown that chemotherapeutic drugs also have immunomodulatory effects. Some chemotherapeutic drugs can enhance anti-tumor immunity by inducing tumor cell immunogenic cell death or performing "immunogenic modulation". In addition, chemotherapeutic drugs can also act on immune cells and even the intestinal flora. A number of clinical trials of chemotherapy combined with immune checkpoint inhibitors are currently underway. This article focuses on the immunomodulatory effect of chemotherapeutic drugs, as well as the potential of chemotherapy combined with ICI on cancer, to provide guidance for the clinical application of chemotherapeutic drugs.

In canonical Wnt/β-catenin signaling pathway, β-catenin/TCF4 (T-cell factor 4) interaction plays an important role in the pathogenesis and development of non-small cell lung cancer (NSCLC), and it is tightly associated with the proliferation, chemoresistance, recurrence and metastasis of NSCLC. Therefore, suppressing β-catenin/TCF4 interaction in Wnt/β-catenin signaling pathway would be a new therapeutic avenue against NSCLC metastasis. In this study, considering the principle of enzyme-linked immunosorbent assay (ELISA), an optimized high-throughput screening (HTS) assay was developed for the discovery of β-catenin/TCF4 interaction antagonists. Subsequently, this ELISA-like screening assay was performed using 2 μg/mL GST-TCF4 βBD and 0.5 μg/mL β-catenin, then a high Z' factor of 0.83 was achieved. A pilot screening of a natural product library using this ELISA-like screening assay identified plumbagin as a potential β-catenin/TCF4 interaction antagonist. Plumbagin remarkably inhibited the proliferation of A549, H1299, MCF7 and SW480 cell lines. More importantly, plumbagin significantly suppressed the β-catenin-responsive transcription in TOPFlash assay. In short, this newly developed ELISA-like screening assay will be vital for the rapid screening of novel Wnt inhibitors targeting β-catenin/TCF4 interaction, and this interaction is a potential anticancer target of plumbagin in vitro.
Carcinoma, Non-Small-Cell Lung ; Cell Line, Tumor ; Enzyme-Linked Immunosorbent Assay ; High-Throughput Screening Assays ; Humans ; Lung Neoplasms ; Transcription Factor 4/genetics* ; beta Catenin/genetics*

The main protease (Mpro) of SARS-CoV-2 is a highly conserved and mutation-resistant coronaviral enzyme, which plays a pivotal role in viral replication, making it an ideal target for the development of novel broad-spectrum anti-coronaviral drugs. In this study, a codon-optimized Mpro gene was cloned into pET-21a and pET-28a expression vectors. The recombinant plasmids were transformed into E. coli Rosetta(DE3) competent cells and the expression conditions were optimized. The highly expressed recombinant proteins, Mpro and Mpro-28, were purified by HisTrapTM chelating column and its proteolytic activity was determined by a fluorescence resonance energy transfer (FRET) assay. The FRET assay showed that Mpro exhibits a desirable proteolytic activity (25 000 U/mg), with Km and kcat values of 11.68 μmol/L and 0.037/s, respectively. The specific activity of Mpro is 25 times that of Mpro-28, a fusion protein carrying a polyhistidine tag at the N and C termini, indicating additional residues at the N terminus of Mpro, but not at the C terminus, are detrimental to its proteolytic activity. The preparation of active SARS-CoV-2 Mpro through codon-optimization strategy might facilitate the development of the rapid screening assays for the discovery of broad-spectrum anti-coronaviral drugs targeting Mpro.
COVID-19 ; Codon/genetics* ; Cysteine Endopeptidases/genetics* ; Escherichia coli/genetics* ; Humans ; Peptide Hydrolases ; SARS-CoV-2 ; Viral Nonstructural Proteins/genetics*