Supplementary Material for: Clinical Outcomes of Prolonged Antiplatelet Therapy after Percutaneous Coronary Intervention in Chronic Kidney Disease Patients: A Systematic Review and Meta-Analysis

Background: The optimal duration of dual antiplatelet therapy (DAPT) after percutaneous coronary intervention (PCI) is not established in chronic kidney disease (CKD) patients. Objective: In this meta-analysis, we evaluated the efficacy and safety of long duration DAPT compared with short duration DAPT in CKD patients after PCI. Methods: We searched PubMed, Cochrane, and Embase to identify studies assessing the effect of DAPT duration in CKD patients with PCI. Endpoints included all-cause mortality, major adverse cardiovascular events (MACE), death or myocardial infarction (MI), revascularization, and bleeding. Event rates were compared with a random-effects model expressed by odds ratio (OR) and 95% confidence interval (CI). Results: Six studies were included. CKD patients with extended DAPT duration were at a lower risk of mortality (OR 1.40, 95% CI: 1.11–1.77), MACE (OR 1.33, 95% CI: 1.17–1.51), mortality or MI (OR 1.24, 95% CI: 1.10–1.40), and stroke (OR 1.28, 95% CI: 1.05–1.56). However, there was no significant difference in revascularization and bleeding events between the two groups. Mortality was higher in patients with dialysis or drug-eluting stent comparing short- to long-term DAPT. Conclusions: Prolonged DAPT might decrease the risk of mortality, MACE, and stroke in patients with CKD without any significant difference in bleeding or revascularization. Additional studies are required to determine whether long-term DAPT could be considered for most CKD patients after PCI

Supplementary Material for: Anti-Caries Effect of Arginine-Containing Formulations in vivo: A Systematic Review and Meta-Analysis

<b><i>Objective:</i></b> To assess the anti-caries effect of arginine-containing formulations in vivo on caries lesions compared with fluorides or placebo. <b><i>Methods:</i></b> Randomized or quasi-randomized human clinical trials wherein arginine was delivered by any method were considered. The MEDLINE, Web of Science, EMBASE, Cochrane Library, and CBM databases were searched to identify relevant articles published up to December 2014. Grey literature was also searched. Two authors performed data extraction independently and in duplicate using data collection forms. Each included study was assessed using the Cochrane risk of bias assessment tool. <b><i>Results:</i></b> Of the 470 studies screened, 31 full articles were scrutinized and assessed for eligibility. Ten studies (n = 15,546 participants) were selected for final inclusion. The meta-analysis results (n = 7 studies) demonstrated a synergistic effect of arginine when used in conjunction with fluoride on early coronal and root caries compared with placebo or fluoride alone. No specific side effects related to arginine usage were identified. <b><i>Conclusions:</i></b> When used in combination with a calcium compound and fluoride, arginine potentially provides a superior anti-caries effect compared with matched formulations of fluoride alone. However, the level of evidence was downgraded because of risks of bias and potential publication bias. In the future, more high quality, non-industry-supported clinical studies in this research area are required before any definitive recommendations can be made

Supplementary Material for: Component-Resolved Diagnosis of Peanut Allergy and Its Possible Origins of Sensitization in China

<b><i>Background:</i></b> Clinical and immunological characteristics of food allergies vary depending on geographic regions. Little is known about peanut allergy in China. The aim of this study was to investigate the peanut sensitization profile in China. <b><i>Methods:</i></b> Thirty-eight participants with immunoglobulin E (IgE)-positive responses to peanuts (peanut-sensitized) were included in our study, and clinical characteristics were evaluated. Total and specific IgE reactivity against peanuts, other plant-derived foods, pollens, and related allergen components were determined. <b><i>Results:</i></b> Eighteen patients were symptomatic when exposed to peanuts. The majority of them presented with systemic reactions. More than half of the peanut-sensitized subjects also suffered from mugwort pollinosis and peach allergy. In patients with both peanut and peach allergies, reactions to peanuts were the same as or severer than those to peaches. Positivity rates of IgE response to rAra h 1-3, 8, and 9 in the peanut allergy group were 5.6, 11.1, 5.6, 22.2, and 83.3%, respectively. 66.7% (12/18) of the peanut-allergic patients were monosensitized to rAra h 9. Anti-nArt v 3 [mugwort nonspecific lipid transfer protein (nsLTP)] IgE positivity in the peanut allergy group was significantly higher than that in the asymptomatic peanut-sensitized group. In Ara h 9 (peanut nsLTP)-sensitized patients with mugwort pollinosis, anti-nArt v 3 IgE levels were remarkably higher than anti-rAra h 9 (peanut nsLTP) IgE levels as well as anti-Pru p 3 (peach nsLTP) IgE levels. <b><i>Conclusions:</i></b> Ara h 9 was the major allergen of peanut, and Ara h 9 monosensitization was the most common peanut sensitization pattern in our population. Furthermore, there was a strong correlation between peanut sensitization and mugwort pollinosis, as well as peach allergy, in our country

Supplementary Material for: Febuxostat Prevents Renal Interstitial Fibrosis by the Activation of BMP-7 Signaling and Inhibition of USAG-1 Expression in Rats

<b><i>Background:</i></b> Renal interstitial fibrosis (RIF) is a common pathology associated with end-stage renal diseases. The activation of bone morphogenetic protein-7 (BMP-7)-Smad1/5/8 pathway seems to alleviate RIF. Uterine sensitization-associated gene-1 (USAG-1), a kidney-specific BMPs antagonist, is associated with the development and prognosis of several renal diseases. Febuxostat is a xanthine oxidase inhibitor that can attenuate the renal dysfunction of patients. The purpose of this study was to investigate the effects of febuxostat on renal fibrosis and to clarify the mechanisms underlying these effects. <b><i>Methods:</i></b> Rats were randomly divided into 6 groups termed a sham-operated group, a unilateral ureteral obstruction (UUO) group, 3 doses of febuxostat groups (low, intermediate and high doses) and a sham group treated with high-dose febuxostat. After 14 days, renal function, relative kidney weight, accumulation of glycogen and collagens were examined by different methods. Expression of α-SMA, transforming growth factor-β1 (TGF-β1), BMP-7 and USAG-1 was detected by western blotting and RT-PCR, respectively. The phosphorylation level of Smad1/5/8 was also quantified by western blotting. <b><i>Results:</i></b> The renal function was declined, and large amounts of glycogen and collagens were deposited in the kidneys of UUO rats compared with the rats in the sham group. Besides, expression of α-SMA and USAG-1 in these kidneys was elevated, and the TGF-β1 was also activated, while the BMP-7-Smad1/5/8 pathway was inhibited. Febuxostat reversed the changes stated earlier, exhibiting protective effects on RIF induced by UUO. <b><i>Conclusion:</i></b> Febuxostat was able to attenuate RIF caused by UUO, which was associated with the activation of BMP-7-Smad1/5/8 pathway and the inhibition of USAG-1 expression in the kidneys of UUO rats

Supplementary Material for: Integrated High Throughput Analysis Identifies GSK3 as a Crucial Determinant of p53-Mediated Apoptosis in Lung Cancer Cells

<strong><em>Background/Aims:</em></strong> p53 dysfunction is frequently observed in lung cancer. Although restoring the tumour suppressor function of p53 is recently approved as a putative strategy for combating cancers, the lack of understanding of the molecular mechanism underlying p53-mediated lung cancer suppression has limited the application of p53-based therapies in lung cancer. <b><i>Methods and Results:</i></b> Using RNA sequencing, we determined the transcriptional profile of human non-small cell lung carcinoma A549 cells after treatment with two p53-activating chemical compounds, nutlin and RITA, which could induce A549 cell cycle arrest and apoptosis, respectively. Bioinformatics analysis of genome-wide gene expression data showed that distinct transcription profiles were induced by nutlin and RITA and 66 pathways were differentially regulated by these two compounds. However, only two of these pathways, 'Adherens junction' and 'Axon guidance', were found to be synthetic lethal with p53 re-activation, as determined via integrated analysis of genome-wide gene expression profile and short hairpin RNA (shRNA) screening. Further functional protein association analysis of significantly regulated genes associated with these two synthetic lethal pathways indicated that GSK3 played a key role in p53-mediated A549 cell apoptosis, and then gene function study was performed, which revealed that GSK3 inhibition promoted p53-mediated A549 cell apoptosis in a p53 post-translational activity-dependent manner. <b><i>Conclusion:</i></b> Our findings provide us with new insights regarding the mechanism by which p53 mediates A549 apoptosis and may cast light on the development of more efficient p53-based strategies for treating lung cancer

Erratum: Endoplasmic Reticulum Stress Predicts Clinical Response to Cyclosporine Treatment in Primary Membranous Nephropathy

<b><i>Background:</i></b> Little is known about the endoplasmic reticulum stress (ERS) marker glucose regulated protein 78 (GRP78) and calcineurin in the kidney in primary membranous nephropathy (PMN) and if they could predict post-cyclosporine treatment outcome. <b><i>Methods:</i></b> This is a retrospective study using a dataset of biopsy-confirmed PMN from Peking Union Medical College Hospital from 1996 to 2014. Seventy-six adult patients treated with cyclosporine as primary immunosuppression for at least 6 months were studied. Immunohistochemistry was used to detect GRP78 and calcineurin in the kidney. Serum calcineurin was assayed by ELISA. Patients were grouped into no-remission (NR, n = 17), partial remission (PR, n = 39), or complete remission (CR, n = 20) at the end of 6 months of treatment. <b><i>Results:</i></b> There was no difference of initial dose of cyclosporine among NR, PR, and CR groups. Kidney calcineurin expression in PMN was significantly increased compared to that in controls (p < 0.0083). The glomerular GRP78 in NR PMN was higher than that in control, CR and PR patients (p < 0.0083). Kidney calcineurin expression and GRP78 expression was positively correlated. However, there were no differences in either serum calcineurin levels or kidney calcineurin expressions among NR, PR or CR groups. There was a negative correlation between serum calcineurin activity and whole kidney calcineurin expression (p = 0.034) or glomerular calcineurin expression (p = 0.007). Neither kidney calcineurin nor GRP78 expression was correlated with proteinuria. <b><i>Conclusions:</i></b> ERS marker GRP78 in the glomeruli but not serum or kidney calcineurin expression could be a useful marker in PMN to negatively predict the response to cyclosporine treatment at the sixth month

Supplementary Material for: Distinguishing intracranial diabetes-related atherosclerotic plaques : A high-resolution MRI-based radiomics study

Introduction: Diabetes markedly affects the formation and development of intracranial atherosclerosis. The study was aimed at evaluating whether radiomics features can help distinguish plaques primarily associated with diabetes. Materials and Methods: We retrospectively analyzed patients who were admitted to our center because of acute ischemic stroke due to intracranial atherosclerosis between 2016 and 2022. Clinical data, blood biomarkers, conventional plaque features and plaque radiomics features were collected for all patients. Odds ratios (ORs) with 95% confidence intervals (CIs) were determined from logistic regression models. The receiver operating characteristic (ROC) curve and area under the ROC curve (AUC) were used to describe diagnostic performance. The DeLong test was used to compare differences between models. Results: Overall, 157 patients (115 men; mean age, 58.7 ± 10.7 years) were enrolled. Multivariate logistic regression analysis showed that plaque length (OR: 1.17; 95% CI: 1.07–1.28) and area (OR: 1.13; 95% CI: 1.02–1.24) were independently associated with diabetes. On combining plaque length and area as a conventional model, the AUCs of the training and validation cohorts for identifying diabetes patients were 0.789 and 0.720, respectively. On combining radiomics features on T1WI and contrast-enhanced T1WI sequences, a better diagnostic value was obtained in the training and validation cohorts (AUC: 0.889 and 0.861). The DeLong test showed the model combining radiomics and conventional plaque features performed better than the conventional model in both cohorts (p < 0.05). Conclusions: The use of radiomics features of intracranial plaques on hrMRI can effectively distinguish culprit plaques with diabetes as the primary pathological cause, which will provide new avenues of research into plaque formation and precise treatment

Supplementary Material for: Association Between Hypoxia-Inducible Factor-2α (HIF-2α) Expression and Colorectal Cancer and Its Prognostic Role: a Systematic Analysis

<b><i>Background/Aims:</i></b> Although some studies showed that HIF-2α expression was correlated with an unfavorable prognosis in colorectal cancer (CRC), the prognostic results remain conflicting in CRC. The present study was performed to evaluate the association between HIF-2α expression and the clinicopathological features of this disease and to examine the potential prognostic role of HIF-2α expression in CRC. <b><i>Methods:</i></b> Pooled odds ratios (ORs) or hazard ratios (HRs) were calculated from available publications, The Cancer Genome Atlas (TCGA) and the Gene Expression Omnibus (GEO) datasets. Trial sequential analysis (TSA) was used to estimate the required sample information. <b><i>Results:</i></b> HIF-2α protein expression was more frequent in CRC than in normal colonic tissues (OR = 150.49, <i>P</i> < 0.001), higher in male than female CRC patients (OR = 1.47, <i>P</i> = 0.008), and lower in high-grade than low-grade CRC (OR = 0.49, <i>P</i> = 0.029). TSA verified the reliability of the above results. HIF-2α expression was not linked to the prognosis of CRC in overall survival (OS), disease-specific survival (DSS), metastasis-free survival, and relapse-free survival, and no significant correlation was found between HIF-2α alteration and OS or disease-free survival (DFS) of CRC. Expression of both HIF-2α and vascular endothelial growth factor (VEGFA, VEGFB, or VEGFC) was associated with a poor metastasis-free survival of CRC (HR = 6.95, HR = 113.51, and HR = 8.11, respectively). No association was observed between HIF-2α expression and DFS in other cancers, but HIF-2α expression was correlated with a worse DFS of CRC (HR = 1.23, <i>P</i> = 0.037). Moreover, HIF-2α expression was linked to a good survival benefit in some cancers (B-cell lymphoma and lung adenocarcinoma: OS, multiple myeloma: DSS, breast cancer: distant metastasis-free survival, liposarcoma: distant recurrence-free survival) (all HRs < 1, <i>Ps</i> < 0.05). <b><i>Conclusions:</i></b> HIF-2α expression may be associated with the carcinogenesis of CRC, which is higher in males than in females, negatively linked to tumor differentiation, and correlated with a worse DFS of CRC. Additional prospective studies are needed

Supplementary Material for: Association of Initial Twice-Weekly Hemodialysis Treatment with Preservation of Residual Kidney Function in ESRD Patients

<b><i>Background:</i></b> Residual kidney function (RKF) has consistently been a predictor of greater survival in maintenance hemodialysis (MHD) patients. The relationship between hemodialysis (HD) treatment frequency and RKF preservation has not been well examined. We hypothesized that initial twice-weekly HD helps in maintaining a longer RKF. <b><i>Methods:</i></b> In a dialysis center in Shanghai, 168 ESRD patients were screened and finally 85 patients were identified for this main cohort study. We first examined these 85 MHD patients; 30 of them were initiated with twice-weekly HD for 6 months or longer and 55 patients were started and maintained on thrice-weekly HD treatment. Then a subcohort study in 48 incident MHD patients was implemented to assess the independent risk factors responsible for RKF decline during the first year of HD therapy. Multivariate logistic regression analysis was then employed to examine the odds ratio of RKF loss. <b><i>Results:</i></b> The main cohort study showed that the clinical outcomes were almost the same between the two groups in 85 patients, but the percent of patients with RKF loss was significantly lower in the twice-weekly group compared with the thrice-weekly group, especially during the first year of HD initiation. In the 48 incident MHD patients, we found no significant differences between the two groups except for variations in the HD frequency, weekly Kt/V. The multivariate analysis showed that factors such as the male gender, HD frequency, URR and intradialytic hypotension episode were associated with RKF loss, and the odds ratio of RKF loss for each additional HD treatment per week was 7.2. <b><i>Conclusion:</i></b> Twice-weekly HD during the first year of dialysis therapy appears to be associated with better RKF preservation

Supplementary Material for: Prenatal Diagnosis of DNA Copy Number Variations by Genomic Single-Nucleotide Polymorphism Array in Fetuses with Congenital Heart Defects

<b><i>Objectives:</i></b> To evaluate the usefulness of single-nucleotide polymorphism (SNP) array for prenatal genetic diagnosis of congenital heart defect (CHD), we used this approach to detect clinically significant copy number variants (CNVs) in fetuses with CHDs. <b><i>Methods:</i></b> A HumanCytoSNP-12 array was used to detect genomic samples obtained from 39 fetuses that exhibited cardiovascular abnormalities on ultrasound and had a normal karyotype. The relationship between CNVs and CHDs was identified by using genotype-phenotype comparisons and searching of chromosomal databases. All clinically significant CNVs were confirmed by real-time PCR. <b><i>Results:</i></b> CNVs were detected in 38/39 (97.4%) fetuses: variants of unknown significance were detected in 2/39 (5.1%), and clinically significant CNVs were identified in 7/39 (17.9%). In 3 of the 7 fetuses with clinically significant CNVs, 3 rare and previously undescribed CNVs were detected, and these CNVs encompassed the CHD candidate genes <i>FLNA</i> (Xq28 dup), <i>BCOR</i> (Xp11.4 dup), and <i>RBL2</i> (16q12.2 del). <b><i>Conclusion:</i></b> Compared with conventional cytogenetic genomics, SNP array analysis provides significantly improved detection of submicroscopic genomic aberrations in pregnancies with CHDs. Based on these results, we propose that genomic SNP array is an effective method which could be used in the prenatal diagnostic test to assist genetic counseling for pregnancies with CHDs