Supplementary Material for: Identifying Gene Signature for the Detection of Ovarian Cancer Based on the Achieved Related Genes

<i>Background:</i> The overall survival rate of ovarian cancer patients is still poor because of the difficulties encountered in detection, diagnosis and treatment. Here, we aim to systematically identify the genetic factors causing ovarian cancer and find the accurate diagnostic and therapeutic targets for ovarian cancer. <i>Methods:</i> We collected the known archived ovarian cancer-related genes from the databases used as the investigated targets and employed the minimum redundancy maximum relevance and random forest classification to identify the novel ovarian cancer-related genes in addition to the known ones. We further identified candidates as the markers for the detection of the ovarian cancer based on the gene expression data and then confirmed them by quantitative real-time PCR. <i>Results:</i> We found out the genetic terms to interpret the mechanism of ovarian cancer. Based on those terms, we predicted 860 novel related genes as candidates. These candidates can act as expression biomarkers for clinical detection and they achieved a 100% accuracy. We verified 10 of them as the optimal biomarkers for detection in the expression data. <i>Conclusion:</i> We employed the features of achieved ovarian cancer-related genes to identify 860 novel ovarian cancer genes. We further validated 10 genes as biomarkers for detection of ovarian cancer

Supplementary Material for: Meta-Analyses of the Effect of <b><i>CYP1A1</i></b> and <b><i>CYP2D6</i></b> Polymorphisms on the Risk of Head and Neck Squamous Cell Carcinoma

<b><i>Background: </i></b><i>CYP1A1</i> and <i>CYP2D6</i> are important genes encoding enzymes involved in the metabolism of toxic chemicals and carcinogens. However, inconclusive results for the association between <i>CYP1A1</i> and <i>CYP2D6</i> polymorphisms and the risk of head and neck squamous cell carcinoma (HNSCC) have been reported. We conducted a meta-analysis to evaluate the association of <i>CYP1A1</i> and <i>CYP2D6</i> polymorphisms with the risk of HNSCC. <b><i>Methods: </i></b>A database search yielded 19 relevant studies. 3 polymorphisms were included in the meta-analysis: <i>CYP1A1</i>, <i>CYP2D6*4</i> and <i>CYP2D6*10</i>. Random or fixed effect models were used in the analysis. <b><i>Results: </i></b>The <i>CYP1A1</i> polymorphism was associated with HNSCC (for m1m1 vs. m1m2: odds ratio (OR) = 1.26, 95% confidence interval (CI) = 1.030-1.542, p_{heterogeneity} = 0.025; for the recessive model: OR = 1.316, 95% CI = 1.065-1.625, p_{heterogeneity} = 0.001). The analysis showed evidence for association between the <i>CYP2D6*4</i> polymorphism and HNSCC in Asian populations; however, negative results were also observed in other models. A significant association was also observed between <i>CYP2D6*10</i> polymorphism and HNSCC risk. <b><i>Conclusions: </i></b>The current study demonstrates that the <i>CYP1A1</i> and CYP2D6 polymorphisms are associated with susceptibility to both development and progression of HNSCC

Erratum: Nornicotine and Nicotine Induced Neovascularization via Increased VEGF/PEDF Ratio

<b><i>Purpose:</i></b> The purpose of the current study was to evaluate the influences of nornicotine and nicotine (NT) in cigarette smoke on the expression of vascular endothelial growth factor (VEGF) and pigment epithelium-derived factor (PEDF) in retinal pigment epithelium cells and human umbilical vein endothelial cells (HUVECs). Furthermore, the angiogenic behaviors of endothelial cells under nornicotine and NT treatment were assessed by using in vitro methods. <b><i>Methods:</i></b> ARPE-19 cells and HUVECs were treated with different concentrations of either nornicotine or NT for different periods of time. The cell proliferative effect was investigated by using the method of MTT analysis. HUVEC migration and tube formation were assessed by using the scratch assay and Matrigel models. The expressions of VEGF and PEDF gene and protein in both types of cells were examined by real-time RT-PCR and Western blot. <b><i>Results:</i></b> There was no proliferation of ARPE-19 cells following treatment with various concentrations of nornicotine or NT. In contrast, nornicotine or NT significantly stimulated HUVEC proliferation, migration and tube formation. Nornicotine and NT upregulated the expression of VEGF but suppressed the expression of PEDF at both mRNA and protein levels in a dose- and time-dependent manner in ARPE-19 cells and HUVECs. <b><i>Conclusions:</i></b> Our results demonstrate that nornicotine and NT promoted endothelial cellular proliferation, migration and tube formation of HUVECs in vitro. These effects might be partly due to simultaneous modulation of VEGF/PEDF signaling in ARPE-19 cells and HUVECs

Supplementary Material for: Risk Factors Associated with Left-Sided Cardiac Valve Calcification: A Case Control Study

<b><i>Objectives:</i></b> To identify risk factors associated with cardiac valve calcification that is easily detectable through routine blood tests in patients who received valve replacement therapy. <b><i>Methods:</i></b> Four hundred patients with valvular heart disease who underwent valve replacement surgery between December 2009 and January 2013 were enrolled in this study. Of these, 77 had valve calcification; the other 323 did not. Multivariate logistic regression analysis was used to assess for risk factors associated with valve calcification. <b><i>Results:</i></b> In our study population, rheumatic valve lesions were the most common reason for valve replacement. Degenerative nonstenotic valve lesion was a protective factor and degenerative stenotic valve lesion was a strong risk factor for valve calcification. Serum levels of gamma-glutamyl transferase (GGT) of between 30 and 46 IU/l and >90 IU/l and total bilirubin (TBIL) of between 15 and 20 μmol/l were positively correlated with valve calcification. Meanwhile, serum calcium (Ca²⁺) levels of between 2.3 and 2.4 mmol/l were negatively correlated with rheumatic valve calcification. <b><i>Conclusions:</i></b> Degenerative stenotic lesion is a risk factor and degenerative nonstenotic lesion a protective factor for cardiac valve calcification. Serum GGT and TBIL levels are positively correlated and serum Ca²⁺ levels negatively correlated with rheumatic cardiac valve calcification

Supplementary Material for: Perilipin1 deficiency prompts lipolysis in lipid droplets and aggravates the pathogenesis of persistent immune activation in Drosophila

Lipid droplets (LDs) are highly dynamic intracellular organelles, which are involved in lots of biological processes. However, the dynamic morphogenesis and functions of intracellular LDs during persistent innate immune responses remain obscure. In this study, we induce long-term systemic immune activation in Drosophila through genetic manipulation. Then, the dynamic pattern of LDs is traced in the Drosophila fat body. We find that deficiency of Plin1, a key regulator of LDs’ reconfiguration, blocks LDs minimization at the initial stage of immune hyperactivation but enhances LDs breakdown at the later stage of sustained immune activation via recruiting the lipase Brummer (Bmm, homologous to human ATGL). The high wasting in LDs shortens the lifespan of flies with high-energy-cost immune hyperactivation. Therefore, these results suggest a critical function of LDs during long-term immune activation and provide a potential treatment for the resolution of persistent inflammation

Supplementary Material for: The Characteristics and Treatment Outcomes of 71 Duodenal Brunner’s gland Adenomas with Endoscopic Submucosal Dissection

Abstract Background: The aim of this study was to investigate outcomes of patients with duodenal Brunner’s gland adenomas (BGAs) that were treated endoscopically. Methods: We identified consecutive 71 patients treated at our center with endoscopic submucosal dissection (ESD) for their duodenal tumors diagnosed pathologically as BGAs over the period between January 1, 2011 and December 31, 2021. We retrospectively analyzed our experience and short- and long-term outcomes of ESD therapy on patients with BGAs. Results: Among 71 BGA patients with an average age of 57 ± 11.7 years (range: 30-82), 48 (67.6%) were male and 23 (32.4%) were female. The accuracy of preoperative diagnosis with endoscopic ultrasonography was 44.0% (22/50). The H. pylori infection was found in 29 patients (29/71, 40.8%). The median size of BGAs was 1.5 cm [interquartile range (IQR) 0.8-2.7cm]. The most common location was the duodenum bulb (50/71, 64.8%). For the ESD procedure, the median operation time was 15.0 minutes (IQR 9.5-25.5 minutes). The en-bloc and the complete resection rates were 97.2% and 92.3%, respectively. ESD-related mild acute obstructive pancreatitis was present in two patients (2/4, 50%) with BGAs located in the ampulla region. During the follow-up period, one patient with a positive peripheral margin experienced tumor recurrence 2 years after the initial ESD. There was no disease-related death for the cohort. Conclusion: ESD was an effective and safe therapeutic option for BGA patients with excellent outcomes. Long-term follow-up is needed

Supplementary Material for: Serum Human epididymis protein 4 as a prognostic predictor of new onset heart failure among women after acute coronary syndrome: a single-center retrospective study

Introduction: Little is known about the prognostic factors among women with acute coronary syndrome (ACS), partly due to the small number of women included in heart failure (HF) clinical trials. Human epididymis protein 4 (HE4) has been proven to be a new biomarker for acute and chronic HF over the years. We hypothesize that HE4 could be a promising predictor. Methods: This retrospective study analyzed data from Zhejiang Provincial People’s Hospital. This study included 302 female patients with ACS between January 1, 2021, and December 1, 2021. The primary outcome was new-onset HF after ACS during the 12-month follow-up period. We used a logistic regression model to evaluate the association between serum HE4 levels and the incidence of HF. Serum HE4 levels were measured at baseline (within 24 hours after admission). Results: Of the 302 female patients, 70 (23.2 %) developed new-onset HF within 12 months. Serum HE4 levels in patients with adverse events were significantly higher than those in patients without events (8.9 (7.3-11.5) pmol/dL vs. 5.9 (5.0-6.8) pmol/dL, P 6.93 pmol/dL distinguished patients at risk of HF with 82.9 % sensitivity and 78.0 % specificity (maximum Youden index J, 0.609). Moreover, HE4 levels were associated with an increased risk of HF. Discussion/Conclusion: We found a strong relationship between HE4 and the occurrence of HF after ACS among women, which might help identify patients at high risk of HF for whom close or intense management should be mandatory

Supplementary Material for: Perilipin1 deficiency prompts lipolysis in lipid droplets and aggravates the pathogenesis of persistent immune activation in Drosophila

Lipid droplets (LDs) are highly dynamic intracellular organelles, which are involved in lots of biological processes. However, the dynamic morphogenesis and functions of intracellular LDs during persistent innate immune responses remain obscure. In this study, we induce long-term systemic immune activation in Drosophila through genetic manipulation. Then, the dynamic pattern of LDs is traced in the Drosophila fat body. We find that deficiency of Plin1, a key regulator of LDs’ reconfiguration, blocks LDs minimization at the initial stage of immune hyperactivation but enhances LDs breakdown at the later stage of sustained immune activation via recruiting the lipase Brummer (Bmm, homologous to human ATGL). The high wasting in LDs shortens the lifespan of flies with high-energy-cost immune hyperactivation. Therefore, these results suggest a critical function of LDs during long-term immune activation and provide a potential treatment for the resolution of persistent inflammation

Supplementary Material for: Inducible GBP5 Mediates the Antiviral Response via Interferon-Related Pathways during Influenza A Virus Infection

<p>Guanylate binding protein (GBP) 5 belongs to the GBP family, which is involved in important cellular processes, including signal transduction, translation, vesicle trafficking, and exocytosis. Structurally, GBPs display a high degree of homology and share highly conserved GTP-binding or hydrolysis domains. GBP5 was reported to be a critical cellular factor in inflammasome assembly. However, little is known about its role in the host antiviral innate immune response. In this study, we found that GBP5 expression was significantly elevated in influenza patients and influenza A virus-infected A549 human lung epithelial cells. The overexpression of GBP5 inhibited virus replication by enhancing the expression of virus-induced interferon (IFN) and IFN-related effectors. Knockdown of GBP5 had the opposite effect. Moreover, GBP5 enhanced endogenous IFN expression by interacting with the NF-κB-essential modulator complex and stimulating NF-κB signaling. Additionally, the expression of proinflammatory factors, such as IL-6, IL-8, tumor necrosis factor-α, cyclooxygenase-2, and inducible nitric oxide synthase, was also activated by GBP5. Taken together, our results reveal that GBP5 inhibited virus replication through the activation of IFN signaling and proinflammatory factors.</p

Supplementary Material for: Dose-Response Relationship between Red Blood Cell Distribution Width and In-Hospital Mortality in Oldest Old Patients with Acute Ischemic Stroke

Introduction: It is crucial to identify predictors of mortality in the early stage of acute ischemic stroke for the oldest old (aged ≥80 years) because of their poor overall survival outcomes. However, limited data are available as the oldest old have often been excluded from previous clinical studies. Hence, we aimed to assess the predictive effect of red blood cell distribution width on in-hospital mortality and the dose-response relationship between the red blood cell distribution width and in-hospital mortality in oldest old with acute ischemic stroke. Methods: A retrospective cohort study was performed in two tertiary hospitals. Patients aged ≥80 years admitted due to acute ischemic stroke from January 1, 2014, to January 31, 2020, were included in the study. We divided the eligible patients into 3 groups with tertiles of red blood cell distribution width. Restrictive cubic spline and robust locally weighted regression analysis were performed to test the dose-response relationship between red blood cell distribution width and the in-hospital mortality risk. All-cause in-hospital mortality was the main study outcome. Results: Overall, 606 patients were included in the final analysis. Red blood cell distribution width was categorized into 3 groups (T1: 15.7%). The rationality of this categorization was then validated with restricted cubic spline and robust locally regression smoothing scatterplot, respectively. After adjusting for demographic and clinical features, a higher red blood cell distribution width was independently associated with in-hospital mortality and the hazard ratio (HR) was 3.31 (95% CI 2.47–4.45, p Conclusions: Red blood cell distribution width may be a valuable and simple measure for predicting in-hospital mortality in oldest old patients with acute ischemic stroke