Gene expression and immune infiltration analysis comparing lesioned and preserved subchondral bone in osteoarthritis.

Background: Osteoarthritis (OA) is a degenerative disease requiring additional research. This study compared gene expression and immune infiltration between lesioned and preserved subchondral bone. The results were validated using multiple tissue datasets and experiments. Methods: Differentially expressed genes (DEGs) between the lesioned and preserved tibial plateaus of OA patients were identified in the GSE51588 dataset. Moreover, functional annotation and protein-protein interaction (PPI) network analyses were performed on the lesioned and preserved sides to explore potential therapeutic targets in OA subchondral bones. In addition, multiple tissues were used to screen coexpressed genes, and the expression levels of identified candidate DEGs in OA were measured by quantitative real-time polymerase chain reaction. Finally, an immune infiltration analysis was conducted. Results: A total of 1,010 DEGs were identified, 423 upregulated and 587 downregulated. The biological process (BP) terms enriched in the upregulated genes included "skeletal system development", "sister chromatid cohesion", and "ossification". Pathways were enriched in "Wnt signaling pathway" and "proteoglycans in cancer". The BP terms enriched in the downregulated genes included "inflammatory response", "xenobiotic metabolic process", and "positive regulation of inflammatory response". The enriched pathways included "neuroactive ligand-receptor interaction" and "AMP-activated protein kinase signaling". JUN, tumor necrosis factor α, and interleukin-1ß were the hub genes in the PPI network. Collagen XI A1 and leucine-rich repeat-containing 15 were screened from multiple datasets and experimentally validated. Immune infiltration analyses showed fewer infiltrating adipocytes and endothelial cells in the lesioned versus preserved samples. Conclusion: Our findings provide valuable information for future studies on the pathogenic mechanism of OA and potential therapeutic and diagnostic targets.

Global, regional, and national temporal trend in burden of major depressive disorder from 1990 to 2019: An analysis of the global burden of disease study.

Major depressive disorder (MDD) is one of the leading causes of disability worldwide. Comprehensive description of the global burden of MDD and its attributable risk factors is essential for policymaking but currently lacking. In this study, we aim to estimate the burden of MDD in terms of incidence, prevalence, and years lived with disability (YLDs), along with its attributable risk factors at global, regional, and rational level between 1990 and 2019, using data from the Global Burden of Diseases, Injuries, and Risk Factors Study 2019. Data analysis was completed on July 1, 2023. In 2019, 274.80 million (95 % uncertainty interval [UI], 241.28 to 312.77) new cases of MDD were identified globally, with an increase of 59 % from 1990. A total of 37.20 million (25.65 to 51.22) YLDs were attributable to MDD, accounting for the largest proportion of mental disorder YLDs (29.7 %). Countries in the low sociodemographic index quantile exhibited the highest age-standardized incidence rate of MDD, with Uganda (7836.2, per 100,000 person-years, 6713.7 to 9181.1) and Palestine (7687.7, 6546.1 to 9023.9) reporting the highest rates among them. The United States had the highest increase in age-standardized rates, with an average annual percent change of 0.99. Females had 1.6 times higher age-standardised rates than males, ranging from 1.2 (Oceania) to 2.2 (tropical Latin America) times across 21 regions. Globally, the proportions of YLDs due to MDD attributable to bullying victimization, childhood sexual abuse, and intimate partner violence were 4.86 %, 5.46 %, and 8.43 % in 2019, respectively. The heavy burden of MDD serves as a stark reminder that a coordinated response from governments and health communities is urgently needed to scale up mental health services and implement effective interventions, particularly in low-income countries.

Automated measurement and grading of knee cartilage thickness: a deep learning-based approach.

Background: Knee cartilage is the most crucial structure in the knee, and the reduction of cartilage thickness is a significant factor in the occurrence and development of osteoarthritis. Measuring cartilage thickness allows for a more accurate assessment of cartilage wear, but this process is relatively time-consuming. Our objectives encompass using various DL methods to segment knee cartilage from MRIs taken with different equipment and parameters, building a DL-based model for measuring and grading knee cartilage, and establishing a standardized database of knee cartilage thickness. Methods: In this retrospective study, we selected a mixed knee MRI dataset consisting of 700 cases from four datasets with varying cartilage thickness. We employed four convolutional neural networks-UNet, UNet++, ResUNet, and TransUNet-to train and segment the mixed dataset, leveraging an extensive array of labeled data for effective supervised learning. Subsequently, we measured and graded the thickness of knee cartilage in 12 regions. Finally, a standard knee cartilage thickness dataset was established using 291 cases with ages ranging from 20 to 45 years and a Kellgren-Lawrence grading of 0. Results: The validation results of network segmentation showed that TransUNet performed the best in the mixed dataset, with an overall dice similarity coefficient of 0.813 and an Intersection over Union of 0.692. The model's mean absolute percentage error for automatic measurement and grading after segmentation was 0.831. The experiment also yielded standard knee cartilage thickness, with an average thickness of 1.98 mm for the femoral cartilage and 2.14 mm for the tibial cartilage. Conclusion: By selecting the best knee cartilage segmentation network, we built a model with a stronger generalization ability to automatically segment, measure, and grade cartilage thickness. This model can assist surgeons in more accurately and efficiently diagnosing changes in patients' cartilage thickness.

Altered neurovascular coupling in the children with attention-deficit/hyperactivity disorder: a comprehensive fMRI analysis.

The coupling between resting-state cerebral blood flow (CBF) and blood oxygenation level-dependent (BOLD) signals reflects the mechanism of neurovascular coupling (NVC), which have not been illustrated in attention-deficit/hyperactivity disorder (ADHD). Fifty ADHD and 42 age- and gender-matched typically developing controls (TDs) were enrolled. The NVC imaging metrics were investigated by exploring the Pearson correlation coefficients between CBF and BOLD-derived quantitative maps (ALFF, fALFF, DCP maps). Three types of NVC metrics (CBF-ALFF, CBF-fALFF, CBF-DCP coupling) were compared between ADHD and TDs group, and the inner association between altered NVC metrics and clinical variables in ADHD group was further analyzed. Compared to TDs, ADHD showed significantly reduced whole-brain CBF-ALFF coupling (P < 0.001). Among regional level (all PFDR < 0.05), ADHD showed significantly lower CBF-ALFF coupling in bilateral thalamus, default-mode network (DMN) involving left anterior cingulate (ACG.L) and right parahippocampal gyrus (PHG.R), execution control network (ECN) involving right middle orbital frontal gyrus (ORBmid.R) and right inferior frontal triangular gyrus (IFGtriang.R), and increased CBF-ALFF coupling in attention network (AN)-related left superior temporal gyrus (STG.L) and somatosensory network (SSN))-related left rolandic operculum (ROL.L). Furthermore, increased CBF-fALFF coupling was found in the visual network (VN)-related left cuneus and negatively correlated with the concentration index of ADHD (R = - 0.299, PFDR = 0.035). Abnormal regional NVC metrics were at widespread neural networks in ADHD, mainly involved in DMN, ECN, SSN, AN, VN and bilateral thalamus. Notably, this study reinforced the insights into the neural basis and pathophysiological mechanism underlying ADHD.

Assessment of the glymphatic function in children with attention-deficit/hyperactivity disorder.

OBJECTIVES: Whether the alternation of the glymphatic system exists in neurodevelopmental disease still remains unclear. In this study, we investigated structural and functional changes in the glymphatic system in the treatment-naïve attention-deficit/hyperactivity disorder (ADHD) children by quantitatively measuring the Virchow-Robin spaces (VRS) volume and diffusion tensor image-analysis along the perivascular space (DTI-ALPS). METHODS: Forty-seven pediatric ADHD patients and 52 age- and gender-matched typically developing (TD) children were recruited in this prospective study. The VRS volume was calculated using a semi-automated approach in axial T2-weighted images. Diffusivities along the x-, y-, and z-axes in the projection, association, and subcortical neural fiber areas were measured. The ALPS index, a ratio that accentuated water diffusion along the perivascular space, was calculated. The Mann-Whitney U test was used to compare the quantitative parameters; Pearson's correlation was used to analyze the correlation with clinical symptoms. RESULTS: The cerebral VRS volume (mean, 15.514 mL vs. 11.702 mL) and the VRS volume ratio in the ADHD group were larger than those in the TD group (all p < 0.001). The diffusivity along the x-axis in association fiber area and ALPS index were significantly smaller in the ADHD group vs. TD group (mean, 1.40 vs.1.59, p < 0.05 after false discovery rate adjustment). Besides, the ALPS index was related to inattention symptoms of ADHD (r = - 0.323, p < 0.05). CONCLUSIONS: Our study suggests that the glymphatic system alternation may participate in the pathogenesis of ADHD, which may be a new research direction for exploring the mechanisms of psycho-behavioral developmental disorders. Moreover, the VRS volume and ALPS index could be used as the metrics for diagnosing ADHD. CLINICAL RELEVANCE STATEMENT: Considering the potential relevance of the glymphatic system for exploring the mechanisms of attention deficit/hyperactivity, the Virchow-Robin spaces volume and the analysis along the perivascular space index could be used as additional metrics for diagnosing the disorder. KEY POINTS: • Increased Virchow-Robin space volume and decreased analysis along the perivascular space index were found in the treatment-naïve attention-deficit/hyperactivity disorder children. • The results of this study indicate that the glymphatic system alternation may have a valuable role in the pathogenesis of attention-deficit/hyperactivity disorder. • The analysis along the perivascular space index is correlated with inattention symptoms of attention-deficit/hyperactivity disorder children.

Room-temperature phosphorescence and fluorescence nanocomposites as a ratiometric chemosensor for high-contrast and selective detection of 2,4,6-trinitrotoluene.

Reports on using complementary colours for high-contrast ratiometric assays are limited to date. In this work, graphitized carbon nitride (g-C3N4) nanosheets and mercaptoethylamine (MEA) capped Mn-doped ZnS QDs were fabricated by liquid exfoliation of bulk g-C3N4, and by a coprecipitation and postmodification strategies, respectively. Mn-doped ZnS quantum dots were deposited onto g-C3N4 nanosheets through an electrostatic self-assembly to form new nanocomposites (denoted as Mn-ZnS QDs@g-C3N4). Mn-ZnS QDs@g-C3N4 can emit a pair of complementary colour light, namely, orange room-temperature phosphorescence (RTP) at 582 nm and blue fluorescence at 450 nm. After 2,4,6-trinitrotoluene (TNT) dosing into Mn-ZnS QDs@g-C3N4 aqueous solution, and pairing with MEA to generate TNT anions capable of quenching the emission of Mn-doped ZnS QDs, the fluorescence colours of the solution changed from orange to blue across white, exhibiting unusual high-contrast fluorescence images. The developed ratiometric chemosensor showed very good linearity in the range of 0-12 µM TNT with a limit of detection of 0.56 µM and an RSD of 6.4 % (n = 5). Also, the ratiometric probe had an excellent selectivity for TNT over other nitroaromatic compounds, which was applied in the ratiometric test paper to image TNT in water, and TNT sensing under phosphorescence mode to efficiently avoid background interference. A high-contrast dual-emission platform for selective ratiometric detection of TNT was therefore established.

Quantification of myelination in children with attention-deficit/hyperactivity disorder: a comparative assessment with synthetic MRI and DTI.

Evaluation of myelin content is crucial for attention-deficit/hyperactivity disorder (ADHD). To estimate myelin content in ADHD based on synthetic MRI-based method and compare it with established diffusion tensor imaging (DTI) method. Fifth-nine ADHD and fifty typically developing (TD) children were recruited. Global and regional myelin content (myelin volume fraction [MVF] and myelin volume [MYV]) were assessed using SyMRI and compared with DTI metrics (fractional anisotropy and mean/radial/axial diffusivity). The relationship between significant MRI parameters and clinical variables were assessed in ADHD. No between-group differences of whole-brain myelin content were found. Compared to TDs, ADHD showed higher mean MVF in bilateral internal capsule, external capsule, corona radiata, and corpus callosum, as well as in left tapetum, left superior fronto-occipital fascicular, and right cingulum (all PFDR-corrected < 0.05). Increased MYV were found in similar regions. Abnormalities of DTI metrics were mainly in bilateral corticospinal tract. Besides, MVF in right retro lenticular part of internal capsule was negatively correlated with cancellation test scores (r = - 0.41, P = 0.002), and MYV in right posterior limb of internal capsule (r = 0.377, P = 0.040) and left superior corona radiata (r = 0.375, P = 0.041) were positively correlated with cancellation test scores in ADHD. Increased myelin content underscored the important pathway of frontostriatal tract, posterior thalamic radiation, and corpus callosum underlying ADHD, which reinforced the insights into myelin quantification and its potential role in pathophysiological mechanism and disease diagnosis. Prospectively registered trials number: ChiCTR2100048109; date: 2021-07.

Disrupted small-world networks in children with drug-naïve atten-tion-deficit/hyperactivity disorder: a DTI-based network analysis.

OBJECTIVES: To explore the alterations in the white matter (WM) structural connectome in children with drug-naïve attention-deficit/hyperactivity disorder (ADHD). METHODS: Forty-nine pediatric ADHD and 51 age- and gender-matched typically developing (TD) children aged 6-14 years old were enrolled. This cross-sectional study applied graph theoretical analysis to assess the white matter organization based on deterministic diffusion tensor imaging (DTI). WM structural connectivity was constructed in 90 cortical and subcor-tical regions, and topological parameters of the resulting graphs were calculated. Networks were compared between two groups. The digit cancellation test (DCT) was taken to evaluate clinical symptom severity in pediatric ADHD, using the concentration index and the total cancellation test scores. Then, a partial correlation analysis was performed to explore the re-lationship between significant topologic metrics and clinical symptom severity. RESULTS: Compared to TDs, ADHD showed an increase in the characteristic path length (Lp), normalized clustering coefficient (γ), small-worldness (σ), and a decrease in the global effi-ciency (Eglob) (all P <0.05). Furthermore, ADHD showed reduced nodal centralities mainly in the regions of default mode (DMN), central executive network (CEN), basal ganglia, and bilateral thalamus (all P <0.05). After performing Benjamini-Hochberg's procedure, only left orbital part of superior frontal gyrus (ORBsup.L) and left caudate (CAU) were statistically significant (P < 0.05, FDR-corrected). In addition, the concentration index of ADHD was negatively correlated with the nodal betweenness of the left orbital part of the middle frontal gyrus (ORBmid.L) (r = -0.302, P = 0.042). CONCLUSIONS: Our findings revealed an ADHD-related shift of WM network topology toward "regularization" pattern, characterized by decreased global network integration, which is also reflected by changed nodal centralities involving DMN, CEN, basal ganglia, and bilateral thalamus. ADHD could be understood by examining the dysfunction of large-scale spatially distributed neural networks.

Trends in the prevalence and treatment of comorbid depression among US adults with and without cancer, 2005-2020.

BACKGROUND: Understanding trend characteristics of depression among cancer survivors is essential for healthcare policies and planning. This study estimates longitudinal trends in the prevalence and treatment of depression among adults in the United States with and without cancer. METHODS: This cross-sectional study focused on adults aged 20 years or older based on nationally representative data from the National Health and Nutrition Examination Surveys 2005-2020. Weighted logistic regression model was established to assess association between depression and cancer status after adjusting various covariates potentially related to depression. RESULTS: Among the 37,283 participants (weighted mean age, 47.5; women, 50.9 %), 3648 (9.8 %) were diagnosed with cancer and 3343 (9.0 %) were screened positive for depression. The age-standardized prevalence of depression showed a U-shaped trend in cancer survivors, decreasing from 11.8 % (95 % confidence interval, 8.4 %-15.2 %) in 2005-2008 to 8.3 % (5.6 %-11.0 %) in 2013-2016, then increasing to 11.7 % (6.3 %-17.2 %) in 2017-2020. These trends varied by population subgroup. Among depressive patients with cancer, antidepressant use increased from 38.6 % (28.7 %-48.5 %) in 2005-2008 to 62.9 % (40.6 %-85.2 %) in 2017-2020, whereas mental health consultation increased slightly. LIMITATIONS: Using a screening questionnaire instead of diagnostic criteria to identify depression; small sample size of patients with cancer; and cross-sectional analysis without prospective outcomes. CONCLUSIONS: From 2005 to 2020, the depression disease burden in patients with cancer eased in 2009-2015, but deteriorated recently. A healthy lifestyle and reasonable treatment for depression, based on an objective examination of depression characteristics, would improve long-term cancer outcomes and quality of life.

Trends in care quality in China from 2011 to 2017: An analysis based on the National Specific (Single) Disease Monitoring System.

Background: The Ministry of Health of China conducted a study targeting in single-disease quality control in 2009, aimed to strengthen quality management and improve health care services. This study retrospectively investigated the trends of quality indicators for six monitored diseases 2011-2017 to evaluate the improvement of care quality for the first batch of single-disease. Methods: We extracted data from the National Specific (Single) Disease Monitoring System for 2011-2017. We focused on six conditions: acute myocardial infarction, heart failure, community-acquired pneumonia, coronary artery bypass graft, hip / knee replacement, and acute ischemic stroke. A total of 56 quality indicators (QIs) were adopted to monitor the quality change and determine the trends in care quality. We also calculated the hospital process composite performance (HPCP) using a denominator-based weighting method for each hospital per year. The estimated annual percentage changes (EAPC) 2011-2017 were calculated at national and regional levels. Results: The results showed that use of four QIs had significant downward trends, whereas 25 QIs (including reversed indicators) showed significant upward trends from 2011 to 2017. The greatest improvement was observed in CAP-4 (antibiotic treatment within four hours after admission to the hospital for critical pneumonia) in the central region (EAPC = 48.36, 95% CI = 15.92-89.87); while the largest decrease appeared in AIS-1 (thrombolytic therapy within 4.5 hours of symptom onset) in the western region (EAPC = -13.44, 95% CI = -24.98,-0.11). An increased HPCP was observed in four diseases nationwide, but not for acute myocardial infarction and heart failure. However, there were significant differences across regions in the process of care and outcomes, with the performance of Eastern and Western regions showing remarkable advantages compared with the Central region. Conclusions: We provide evidence for major advancement in care quality in China nationwide. However, the improvement of care in China was unbalanced geographically and should be carefully considered. Future challenges include expanding the coverage of quality monitoring, greater delivery efficiency, and region-balanced health care.

The quality disparity of stroke care over time: An analysis based on the national dataset from 2011 to 2017.

BACKGROUND: Adherence to evidence-based hospital stroke care is variable and may change over time. It is important to determine which process measures are associated with variation in outcome. In a large dataset, we analyzed the association between process and outcome and the fluctuations of indicators over time, and identified quality indicators (QIs) that should be prioritized for improving the quality of stroke care. METHODS: We analyzed data from 123,259 patients diagnosed with acute ischemic stroke (AIS) who were treated at 109 large tertiary hospitals in China between January 2011 and May 2017. In total, 12 stroke treatment indicators were selected to calculate the hospital process composite performance (HPCP). Hospitals were divided into subgroups according to the time trend of HPCP estimated by the Group-Based Model. We analyzed the influence of hospital subgroups on the patient outcomes using a multi-level model and explored the QIs that led to variation. RESULTS: The HPCP trends for stroke indicators of 109 hospitals over 7 years were divided into two groups (Group 1, low-HPCP; Group 2, high-HPCP). After adjusting for patient age, medical insurance, comorbidities, patterns of admission, and NIHSS-scores, patients in the high-HPCP group presented higher rate of independence and longer length of stay compared to the low-HPCP group. The multi-level model showed that there was a statistically significant difference in the utilization rate between the two groups, with most marked differences seen in emergency assessment and function evaluation indicators. CONCLUSION: Variation in the quality of stroke care exists across hospitals, and better adherence to guideline-based care is associated with improved outcomes. We found that QIs related to emergency examination and functional assessment were the main factors differing between good and poor adherers to stroke indicators, suggesting that quality improvement in stroke care could prioritize these QIs.

Gene Variants in Two Families with Inherited Coagulation Factor XI Deficiency and Identification of Mutations.

INTRODUCTION: Mutations in the F11 gene can cause factor XI (FXI) deficiency, leading to abnormal coagulation activity and injury-related bleeding tendency. Therefore, identifying F11 gene mutations and studying the molecular basis will help us understand the pathogenesis of FXI deficiency. METHODS: Coagulation tests and gene sequencing analysis of all members were performed. FXI wild-type and mutant expression plasmids were constructed and transfected into HEK293FT cells. The FXI protein expression level was evaluated by ELISA and Western blot. RESULTS: The FXI activity (FXI:C) and FXI antigen (FXI:Ag) of proband-1 were decreased to 2% and 5%, respectively. FXI:C and FXI:Ag of proband-2 were reduced to 15% and 32%, respectively. Four mutations were found in the two unrelated families, including c.536C>T (p.T179M), c.1556G>A (p.W519*), c.434A>G (p.H145R), and c.1325_1325delT (p.L442Cfs*8). In vitro studies in transiently transfected HEK293FT cells demonstrated that p.T179M, p.W519*, and p.L442Cfs*8 mutations significantly lowered the FXI levels in the culture media. The FXI levels in the culture media and cell lysates of p.H145R mutation were similar to the wild type. CONCLUSION: Our results confirm that the four mutations in the F11 gene are causative in the 2 FXI deficiency families. Moreover, the p.H145R mutation is a cross-reactive material (CRM)-positive phenotype. The other three mutations are CRM-negative phenotypes and lead to FXI protein secretion disorder.

A Novel Homozygous Missense Mutation (Ile583Asn) in a Consanguineous Marriage Family with Hereditary Factor XII Deficiency: A Case Report.

BACKGROUND: Hereditary coagulation factor XII (FXII) deficiency is an autosomal recessive disorder. At present, the contribution of severe FXII deficiency to the development of thromboembolism is still undetermined. There are limited reports on the relationship between the FXII defect and thromboembolism. CASE PRESENTATION: A 27-year-old woman came to our hospital for the treatment of shoulder trauma and cervical disc herniation caused by a car accident. The shoulder trauma was treated with five stitches. After physical examination, imaging examination, and routine coagulation examination, cervical disc herniation was treated conservatively. Combined with the examination results, the patient was diagnosed with FXII deficiency. Unfortunately, the patient was readmitted 10 days after the trauma with edema in the lower limbs and secondary varicose veins. The D-dimer increased to 6.22 mg/L. Thrombus in the inferior vena cava and right common iliac was shown by lower limb venography. According to the patient's medical history, the F12 gene was analyzed by direct sequencing. The patient was also screened for other thrombotic risk factors. Genetic analysis showed that the patient had a c.1748T > A (p.Ile583Asn) homozygous missense mutation in exon 14 of the F12 gene. No other hereditary thrombophilia risk factors screened were positive in the patient. CONCLUSION: The p.Ile583Asn missense mutation in exon 14 of the F12 gene might be responsible for the reduction of the FXII level in the patient.

Evaluation of individual-based morphological brain network alterations in children with attention-deficit/hyperactivity disorder: a multi-method investigation.

To investigate the topological organization of individual-based morphological brain networks (MBNs) in attention-deficit/hyperactivity disorder (ADHD) children with different methods. A total of 60 ADHD children and 60 typically developing (TD) controls matched for age and gender were enrolled. Each participant underwent a structural 3D T1-weighted scan. Based on the inter-regional morphological similarity of GM regions, Kullback-Leibler-based similarity (KLS), Multivariate Euclidean Distance (MED), and Tijms's method were used to construct individual-based MBNs, respectively. The between-group difference of global and nodal network topological profiles was estimated, and partial correlation analysis was used for further analysis. According to KLS and MED-based network, ADHD showed a decreased global efficiency (Eglob) and increased characteristic path length (Lp) compared to the TD group, while Tijms's method-based network showed no between-group difference in global and nodal profiles. Nodal profiles were significantly decreased in the bilateral caudate, and nodal efficiency of the bilateral caudate was negatively correlated with clinical symptom severity of ADHD (P < 0.05, FDR-corrected) by the KLS-based network. Nodal betweenness was significantly decreased in the left inferior occipital gyrus and correlated with clinical symptom severity of ADHD (P < 0.05, FDR-corrected) by the MED-based network. ADHD was found to have a significantly less integrated organization and a shift to a "weaker small-worldness" pattern, while abnormal nodal profiles were mainly in the corpus striatum and default-mode networks. Our study highlights the crucial role of abnormal morphological connectivity patterns in understanding the brain maturational effects in ADHD and enriching the insights into MBNs at an individual level.

[Analysis of the molecular pathogenesis of hereditary protein C deficiency due to a p.Gly86Asp variant of the PROC gene].

OBJECTIVE: To explore the molecular pathogenesis of hereditary protein C (PC) deficiency due to a p.Gly86Asp variant of the PROC gene through in vitro expression experiment. METHODS: Wild type and Gly86Asp mutant expression plasmids of PC were constructed and respectively transfected into HEK 293FT cells. Total RNA was extracted from the transfected cells, and the expression of PROC gene was determined by quantitative real-time PCR (qRT-PCR). PC antigen (PC:Ag) in the supernatant of cell culture and cell lysate was determined by enzyme-linked immunosorbent assay (ELISA), and the level of PC protein was detected by Western blotting. RESULTS: qRT-PCR has detected no significant difference in the transcription level of wild-type and mutant-type PC. Compared with the wild type, the level of mutant PC:Ag in the supernatant and cell lysate were 81.3%±2.6% and 110.0%±2.8%, respectively. No difference was detected in the molecular weight between the wild-type and mutant-type PC by Western blotting. The PC content of mutant type was higher than wild-type in cell lysate, while the opposite was found with the cell culture supernatant. CONCLUSION: The impaired secretion by mutant PC may be the molecular mechanism of PC deficiency caused by the p.Gly86Asp variant.

A Novel and Robust Prognostic Model for Hepatocellular Carcinoma Based on Enhancer RNAs-Regulated Genes.

Evidence has demonstrated that enhancer RNAs (eRNAs) play a vital role in the progression and prognosis of cancers, but few studies have focused on the prognostic ability of eRNA-regulated genes (eRGs) for hepatocellular carcinoma (HCC). Using gene expression profiles of HCC patients from the TCGA-LIHC and eRNA expression profiles from the enhancer RNA in cancers (eRic) data portal, we developed a novel and robust prognostic signature composed of 10 eRGs based on Lasso-penalized Cox regression analysis. According to the signature, HCC patients were stratified into high- and low-risk groups, which have been shown to have significant differences in tumor immune microenvironment, immune checkpoints, HLA-related genes, DNA damage repair-related genes, Gene-set variation analysis (GSVA), and the lower half-maximal inhibitory concentration (IC50) of Sorafenib. The prognostic nomogram combining the signature, age, and TNM stage had good predictive ability in the training set (TCGA-LIHC) with the concordance index (C-index) of 0.73 and the AUCs for 1-, 3-, and 5-year OS of 0.82, 0.77, 0.74, respectively. In external validation set (GSE14520), the nomogram also performed well with the C-index of 0.71 and the AUCs for 1-, 3-, and 5-year OS of 0.74, 0.77, 0.74, respectively. In addition, an important eRG (AKR1C3) was validated using two HCC cell lines (Huh7 and MHCC-LM3) in vitro, and the results demonstrated the overexpression of AKR1C3 is related to cell proliferation, migration, and invasion in HCC. Altogether, our eRGs signature and nomogram can predict prognosis accurately and conveniently, facilitate individualized treatment, and improve prognosis for HCC patients.

Development and Validation of a Novel Prognostic Model for Lower-Grade Glioma Based on Enhancer RNA-Regulated Prognostic Genes.

Enhancer RNAs (eRNAs) are present specifically in tumors, where they affect the expression of eRNA-regulated genes (ERGs). Owing to this characteristic, ERGs were hypothesized to improve prognosis of overall survival in heterogeneous low-grade and intermediate-grade gliomas. This study aimed to construct and validate an ERG prognostic tool to facilitate clinical management, and offer more effective diagnostic and therapeutic biomarkers for glioma. Survival-related eRNAs were identified, and their ERGs were selected based on eRNA and target gene information. The ERG prognostic model was constructed and validated using internal and external validation cohorts. Finally, biological differences related to the ERG signature were analysed to explore the potential mechanisms influencing survival outcomes. Thirteen ERGs were identified and used to build an ERG risk signature, which included five super-enhancer RNA (seRNA)-regulated genes and five LGG-specific eRNA-regulated genes. The prognostic nomogram established based on combining the ERG score, age, and sex was evaluated by calibration curves, clinical utility, Harrell's concordance index (0.86; 95% CI: 0.83-0.90), and time-dependent receiver operator characteristic curves. We also explored potential immune-related mechanisms that might cause variation in survival. The established prognostic model displayed high validity and robustness. Several immune-related genes regulated by seRNAs or specific eRNAs were identified, indicating that these transcripts or their genes were potential targets for improving immunotherapeutic/therapeutic outcomes. The functions of an important specific eRNA-regulated gene (USP28) were validated in robust vitro experiments. In addition, the ERG risk signature was significantly associated with the immune microenvironment and other immune-related features.