Supplementary Material for: Unraveling the Expression Profiles of Long Noncoding RNAs in Rat Cardiac Hypertrophy and Functions of lncRNA BC088254 in Cardiac Hypertrophy Induced by Transverse Aortic Constriction

<p>Long noncoding RNAs (lncRNAs), although initially considered as genomic transcription noise, have been demonstrated to play pivotal roles in multiple biological processes and are increasingly recognized as contributors to the pathology of cancer, neurodegenerative diseases, diabetes, heart diseases, and inflammation. However, studies on the roles of lncRNAs in angiocardiopathy, particularly in cardiac hypertrophy, are still preliminary. In our study, differentially expressed lncRNAs in rat cardiac hypertrophy induced by transverse aortic constriction (TAC) were identified by microarray analysis and validated using quantitative real-time polymerase chain reaction (RT-PCR). Briefly, we identified 6,969 lncRNAs, among which 80 lncRNAs were significantly upregulated and 172 lncRNAs were significantly downregulated. Quantitative RT-PCR was used to validate the differential expression of 5 lncRNAs in myocardial tissue RNA. Further, pathway analysis indicated that 25 pathways corresponded to upregulated transcripts and 20 pathways corresponded to downregulated transcripts. Third, by coexpression network analysis, we found a correlation between BC088254 and phb2 (prohibitin 2) and verified this expression by RT-PCR and Western blot. This is the first study to reveal differentially expressed lncRNAs in rat cardiac hypertrophy induced by TAC, indicating potential lncRNA mechanisms of action in myocardial hypertrophy. We also found that lncRNA BC088254 may have a certain role in myocardial hypertrophy induced by TAC and functional relevance between lncRNA BCO88254 and phb2, but the relationship between these two factors is unclear.</p

Supplementary Material for: Primary Malignant Melanoma of the Endometrium: A Case Report

Introduction: Malignant melanoma most commonly occurs in the skin. Primary malignant melanoma of endometrium is quite rare. Its diagnosis depends on clinical characteristics and pathological examination. It usually exhibits high degree of tumor histology, early onset of distant metastases, and unfavorable prognoses. Case Presentation: In this paper, we report a case of a 73-year-old woman with primary malignant melanoma of endometrium. This patient denied a history of nevus removal or any family medical history of cancer. She was admitted to the hospital for irregular vaginal bleeding after menopause and performed an endometrial biopsy. Pathological of the scrapings suggested malignant melanoma. She subsequently underwent a radical surgery. The final pathology diagnosis was primary malignant melanoma of endometrium, and BRAF gene mutation was detected. The tumor staged as IVB according to the International Federation of Gynecology and Obstetrics (FIGO) classification. Thus, she then started adjuvant chemotherapy. This patient is currently on oral targeted therapy and is still being followed up. Conclusion: Mucosal melanoma is infrequent, and primary malignant melanoma of endometrial is a rare subtype. To the best of our knowledge, malignant melanoma originating from endometrium has never been reported before. It has a high degree of malignancy and is prone to early metastasis. Further investigations are warranted to explore its underlying pathogenesis, management, and outcomes

Supplementary Material for: VEGF-A Inhibition Ameliorates Podocyte Apoptosis via Repression of Activating Protein 1 in Diabetes

<b><i>Background/Aims:</i></b> Vascular endothelial growth factor-A (VEGF-A) upregulation and podocyte apoptosis have been documented in diabetes. This study was designed to investigate whether inhibiting VEGF-A could ameliorate podocyte apoptosis in diabetes and the underlying mechanisms. <b><i>Methods:</i></b> In vitro, small interfering RNAs (siRNAs) of VEGF-A and activator protein 1 (AP-1, c-fos and c-jun), bevacizumab (VEGF-A inhibitor) and SP600125 (AP-1 inhibitor) were added to high glucose (30 mM) induced podocytes. Luciferase reporter assay was used to investigate whether AP-1 was a direct target of VEGF-A. In vivo, bevacizumab and SP600125 were administered to 12-week-old streptozotocin-induced male Sprague Dawley rats. The level of VEGF-A, c-fos, c-jun and bcl-2 were examined using immunostaining and Western blot analysis. Podocyte apoptosis was detected using the terminal deoxynucleotidyl transferase-mediated uridine 5′-triphosphate-biotin nick end labeling (TUNEL) assay, electron microscopy and flow cytometry. <b><i>Results:</i></b> Silencing VEGF-A or AP-1 upregulated bcl-2 and ameliorated podocyte apoptosis. Silencing VEGF-A decreased the level of c-fos and c-jun and bevacizumab and SP600125 treatment attenuated podocyte apoptosis. Luciferase reporter activity of VEGF-A-3′-UTR constructs was significantly provoked when stimulated with TGF-β1. In diabetic rat kidneys, VEGF-A co-localized with bcl-2 in podocytes. With bevacizumab and SP600125 treatment, the level of VEGF-A and AP-1 decreased while bcl-2 increased. Podocyte apoptotic rate was reduced with condensed podocyte nuclei less frequently observed. The urine albumin excretion rate (UAER) and albumin/creatinine were improved. <b><i>Conclusion:</i></b> This study demonstrates VEGF-A inhibition ameliorates podocyte apoptosis by regulating AP-1 and bcl-2 signaling. AP-1 is a direct target of VEGF-A and a novel player in podocyte apoptosis

Supplementary Material for: Injection of hTERT-Transduced Endothelial Progenitor Cells Promotes Beneficial Aortic Changes in a High-Fat Dietary Model of Early Atherosclerosis

<i>Objectives:</i> Cultured endothelial progenitor cells (EPCs) display troubling issues that adversely affect their applicability to endothelial regeneration. We hypothesized that transduction of the human telomerase catalytic subunit (hTERT) gene would enhance EPC function in treating dietary-induced early atherosclerosis (AS). <i>Methods:</i> A dietary-induced early AS model was successfully constructed in 90 healthy male rats, while 30 healthy control (HC) rats were normally fed. Four experimental groups were constructed: an untreated HC group; an untreated AS group injected with PBS; a null EPC AS group injected with null vector-transduced EPCs, and an hTERT EPC AS group injected with hTERT-transduced EPCs. Two months postinjection, abdominal aortas were extracted to validate EPC integration and comparatively assess mRNA and protein expression of the early atherosclerotic markers VCAM-1, ICAM-1, LFA-1, Mac-1, CD44, MCP-1, endothelial nitric oxide synthase (eNOS), and apolipoprotein E. <i>Results:</i> In vitro, hTERT transduction of EPCs resulted in a significantly superior proliferative capacity as well as significantly higher NO, iNOS, and LDH secretory capacity. In vivo injection of hTERT-transduced EPCs produced significant reductions in CD44 and MCP-1 expression as well as a significant increase in eNOS expression relative to injection with null vector-transduced EPCs (all p < 0.05). <i>Conclusion:</i>hTERT-transduced human EPCs may be useful in treating dietary-induced early AS

Supplementary Material for: A Bayesian Partitioning Model for the Detection of Multilocus Effects in Case-Control Studies

<b><i>Background:</i></b> Genome-wide association studies (GWASs) have identified hundreds of genetic variants associated with complex diseases, but these variants appear to explain very little of the disease heritability. The typical single-locus association analysis in a GWAS fails to detect variants with small effect sizes and to capture higher-order interaction among these variants. Multilocus association analysis provides a powerful alternative by jointly modeling the variants within a gene or a pathway and by reducing the burden of multiple hypothesis testing in a GWAS. <b><i>Methods:</i></b> Here, we propose a powerful and flexible dimension reduction approach to model multilocus association. We use a Bayesian partitioning model which clusters SNPs according to their direction of association, models higher-order interactions using a flexible scoring scheme and uses posterior marginal probabilities to detect association between the SNP set and the disease. <b><i>Results:</i></b> We illustrate our method using extensive simulation studies and applying it to detect multilocus interaction in Atherosclerosis Risk in Communities (ARIC) GWAS with type 2 diabetes. <b><i>Conclusion:</i></b> We demonstrate that our approach has better power to detect multilocus interactions than several existing approaches. When applied to the ARIC study dataset with 9,328 individuals to study gene-based associations for type 2 diabetes, our method identified some novel variants not detected by conventional single-locus association analyses

Supplementary Material for: Infantile Hemangioma-Like Vascular Lesion in a 26-Year-Old Woman after Abortion

A 26-year-old woman (G2P1A1) presented with a 5-week history of multiple red marks on her body after a therapeutic abortion. A physical examination found 15 palpable red marks on her head, neck, chest, arms and legs. Proliferating endothelial cells, which expressed CD31, CD34, von Willebrand factor, but not Glut-1 and merosin, were observed in the lesional area by histopathological analyses. Histocompatibility antigen typing of 2 lesions was identical to a sample from peripheral blood. Accelerated regression was observed in 2 lesions treated by intralesional injection of betamethasone, while spontaneous regression was observed within 9 months in the remaining lesions without any treatment. Rapid growth, spontaneous regression and histological analyses in this case support the diagnosis of ‘infantile hemangioma-like vascular lesion'

Supplementary Material for: Initiation and Cessation Timing of Renal Replacement Therapy in Patients with Type 1 Cardiorenal Syndrome: An Observational Study

<p><b><i>Background/Aims:</i></b> Renal replacement therapy (RRT) is a rescue therapy for patients with type 1 cardiorenal syndrome (CRS) with poor prognoses. However, the optimal timing for initiation and cessation of RRT remains controversial. The purpose of this study was to determine the optimal timing of initiation and cessation of RRT for patients with type 1 CRS. <b><i>Methods:</i></b> In this retrospective analysis, patients with refractory type 1 CRS receiving RRT were divided into 3 groups according to weaning from RRT and death within 90 days. Baseline characteristics, underlying heart disease, comorbidities, drug use before RRT, indicators of RRT initiation, and prognosis were compared between the 3 groups. <b><i>Results:</i></b> Fifty-two patients were enrolled, which included 27 males and 25 females with a mean age of 70.7 ± 16.1 years and a 90-day mortality rate of 65.4%. The mean urine output before RRT initiation was 800 mL/ 24 h in the RRT-independent group, 650 mL/24 h in the RRT-dependent group, and 345 mL/ 24 h in the death group (<i>p</i> = 0.021). Additionally, there were obvious differences in fluid balance between the 3 groups (167, 250, and 1,270 mL, respectively, <i>p</i> = 0.016). Patients could be successfully weaned from RRT when urine output was >880 mL and fluid balance volume was <150 mL. <b><i>Conclusion:</i></b> The mean fluid balance of survivors was remarkably less than that of the death group at RRT initiation. RRT termination can be considered when urine output is >880 mL/24 h and volume balance is <150 mL/24 h.</p

Supplementary Material for: Blood eosinophil count and its determinants in a Chinese population-based cohort

Introduction: Blood eosinophil count has been shown markedly variable across different populations. However, its distribution in Chinese general population remains unclear. We aimed to investigate blood eosinophil count and its determinants in a Chinese general population. Methods: In this population-based study, general citizens of Sichuan province in China were extracted from the China Pulmonary Health study. Data on demographics, personal and family history, living condition, lifestyle, spirometry and complete blood count test were obtained and analyzed. A stepwise multivariate binary logistic regression analysis was performed to identify determinants of high blood eosinophils (>75th percentile). Results: A total of 3310 participants were included, with a mean age (SD) of 47.0 (15.6) years. In total population, the median blood eosinophil count was 110.0 (IQR 67.2-192.9) cells/μL, lower than that in smokers (133.4 cells/μL, IQR 79.3-228.4) and patients with asthma (140.7 cells/μL, IQR 79.6-218.2) or post-bronchodilator airflow limitation (141.5 cells/μL, IQR 82.6-230.1), with a right-skewed distribution. Multivariate analyses revealed that oldness (aged ≥60 years) (OR 1.66, 95% CI 1.11-2.48), smoking ≥ 20 pack-years (OR 1.90, 95% CI 1.20-3.00), raising dog/cat (OR 1.72, 95% CI 1.17-2.52) and occupational exposure to dust, allergen and harmful gas (OR 1.58, 95% CI 1.15-2.15) were significantly associated with high blood eosinophils. Conclusion: This study identifies a median blood eosinophil count of 110.0 cells/μL and determinants of high blood eosinophils in a Chinese general population, including oldness (aged ≥60 years), smoking ≥ 20 pack-years, raising dog/cat and occupational exposure to dust, allergen and harmful gas

Supplementary Material for: Association between waist-hip ratio and Female Infertility in the United States: Data from National Health and Nutrition Examination Survey 2017–2020

Introduction: Obesity has previously been correlated with an elevated risk of reproductive system diseases in women. The waist-hip ratio (WHR) has been shown to be correlated with visceral fat, making it one of the most commonly used indicators of abdominal obesity. However, little is known about the relationship between WHR and infertility. Therefore, the aim of this study was to evaluate the effect of the WHR on infertility in women of childbearing age. Methods: The study used cross-sectional data from women aged 20 to 45 who participated in the National Health and Nutrition Examination Survey (NHANES), which was conducted between 2017 and 2020. We collected details of their Waist Circumference, hip Circumference, fertility status and several other essential variables. We used multivariate logistic regression analysis and subgroup analyses to assess the association between WHR and infertility. Results: There were 976 participants, with 12.0% (117/976) who experienced infertility. After adjusting for potential confounding factors, our multivariate logistic regression analysis revealed that every 0.1 unit increase in WHR resulted in a more than 35% higher risk of infertility (OR (95% CI): 1.35 (1.01~1.81), P = 0.043). Compared to the group with WHR <0.85, the risk of infertility increased in the group with WHR ≥0.85, with an adjusted odds ratio (OR) of 1.74 (95% CI 1.06~2.85). When WHR was treated as a continuous variable, it was observed that each 0.1 unit increase in WHR was associated with a relatively high risk in the secondary infertility population after adjusting all covariates, with an OR of 1.66 (95% CI 1.14~2.40, P = 0.01). When WHR was analyzed as a categorical variable, the group with WHR≥0.85 exhibited a significantly higher risk of secondary infertility than the group with WHR<0.85, with the OR of 2.75 (95% CI: 1.35-5.59, P = 0.01) after adjusting for all covariates. Furthermore, the interaction analysis indicated that there was a significant interaction between age status on WHR and the risk of infertility. Conclusion: WHR showed a positive correlation with the risk of infertility. This study highlights the importance of effectively managing abdominal fat and promoting the maintenance of optimal WHR levels to mitigate the progression of infertility, particularly for younger women

Supplementary Material for: Rapid initiation of peritoneal dialysis by automated peritoneal dialysis or hemodialysis: a randomized clinical trial

Introduction: It is still controversial whether automated peritoneal dialysis (APD) or hemodialysis (HD) is a more favorable choice for the rapid initiation of peritoneal dialysis. Methods: A pilot randomized prospective controlled trial was carried out in Shanghai Ruijin Hospital. Sixty-seven patients who chose long-term peritoneal dialysis treatment and needed unplanned dialysis were enrolled and randomized into HD-CAPD group (33 cases) or APD-CAPD group (34 cases) based on the dialysis modality during the transition period (within 14 days from the day PD catheter was implanted). Continuous ambulatory peritoneal dialysis started after the transition period. The primary outcome was the decline rates of residual glomerular filtration rate (GFR). Secondary outcomes included the rates of mechanical complications, the rates of infectious complications and complications of ESRD. Results: We found residual GFR decline were faster in HD-CAPD group than in APD-CAPD group (0.06 ml/min/w vs 0.03ml/min/w, P<0.01). The incidences of mechanical complications were similar in APD-CAPD group comparing with HD-CAPD group, including hernia (2.9% vs 3.0%, P=1.00), catheter malposition (0.02 episodes/patient-months vs 0.02 episodes/patient-months, P=0.70), leakage (5.9% vs 6.1%, P=1.00) and omental wrap (0 episode vs 3 episodes, P=0.368). Though the one-year overall infection rates were similar (0.03 episodes/patient-months vs 0.05 episodes/patient-months, P=0.10), APD-CAPD group had lower rate of bacteremia compared to HD-CAPD group (0 episodes/patient-months vs 0.02 episodes/patient-months, P<0.01). Conclusions: Both APD and HD could be used for patients who need to start dialysis in an unplanned manner. APD may have the advantage in protecting residual renal functions among these patients