Supplementary Material for: Does BDNF Val66Met Polymorphism Confer Risk for Posttraumatic Stress Disorder?

<p><b><i>Background:</i></b> Evidence has indicated that <i>BDNF</i> (brain-derived neurotrophic factor) Val66Met genetic variant could be linked to the development of posttraumatic stress disorder (PTSD). However, clinical observations studying the <i>BDNF</i> polymorphism and the risk of PTSD yielded contradictory results. In this meta-analysis we evaluated the association between <i>BDNF</i> Val66Met and PTSD risk. <b><i>Method:</i></b> Systematic searches in online databases retrieved 6 relevant publications. Different inherited models were utilized to perform the pooled analysis. Subgroup analyses and sensitive analyses based on Hardy-Weinberg equilibrium (HWE) test results were also carried out. <b><i>Results:</i></b> Our study did not found a significant overall effect of <i>BDNF</i> Val66Met on the susceptibility to PTSD under various genetic models. In contrast, subgroup analyses suggested that the stress status of the control group, but not ethnicity, may influence the relationship of <i>BDNF</i> Val66Met with PTSD risk. After the exclusion of a study that was not in HWE, our conclusions remained unchanged during the influence analyses. <b><i>Conclusions:</i></b> This meta-analysis suggested no genetic association of <i>BDNF</i> Val66Met with vulnerability to PTSD. Further research studies are warranted to clarify the impact of <i>BDNF </i>variants on the occurrence of PTSD.</p

Supplementary Material for: Palmitic Acid Curcumin Ester Facilitates Protection of Neuroblastoma against Oligomeric Aβ₄₀ Insult

<b><i>Background/Aims:</i></b> The generation of reactive oxygen species (ROS) caused by amyloid-β (Aβ) is considered to be one of mechanisms underlying the development of Alzheimer’s disease. Curcumin can attenuate Aβ-induced neurotoxicity through ROS scavenging, but the protective effect of intracellular curcumin on neurocyte membranes against extracellular Aβ may be compromised. To address this issue, we synthesized a palmitic acid curcumin ester (P-curcumin) which can be cultivated on the cell membrane and investigated the neuroprotective effect of P-curcumin and its interaction with Aβ. <b><i>Methods:</i></b> P-curcumin was prepared through chemical synthesis. Its structure was determined via nuclear magnetic resonance (NMR) and high-resolution mass spectrometry (HRMS). An MTT assay was used to assess Aβ cytotoxicity and the protective effect of P-curcumin on SH-SY5Y cells. The effect of P-curcumin on Aβ-induced ROS production <i>in vitro</i> and <i>in vivo</i> were assessed based on changes in dichlorofluorescein (DCF) fluorescence. A spectrophotometric method was employed to detect lipid peroxidation. To mimic the interaction of P-curcumin on cell membranes with Aβ, liposomes were prepared by thin film method. Finally, the interactions between free P-curcumin and P-curcumin cultivated on liposomes and Aβ were determined via spectrophotometry. <b><i>Results:</i></b> A novel derivative, palmitic acid curcumin ester was prepared and characterized. This curcumin, cultivated on the membranes of neurocytes, may prevent Aβ-mediated ROS production and may inhibit the direct interaction between Aβ and the cellular membrane. Furthermore, P-curcumin could scavenge Aβ-mediated ROS as curcumin <i>in vitro</i> and <i>in vivo</i>, and had the potential to prevent lipid peroxidation. Morphological analyses showed that P-curcumin was better than curcumin at protecting cell shape. To examine P-curcumin’s ability to attenuate direct interaction between Aβ and cell membranes, the binding affinity of Aβ to curcumin and P-curcumin was determined. The association constants for free P-curcumin and curcumin were 7.66 × 10⁴ M^-1 and 7.61 × 10⁵ M^-1, respectively. In the liposome-trapped state, the association constants were 3.71 × 10⁵ M^-1 for P-curcumin and 1.44× 10⁶ M^-1 for curcumin. With this data, the thermodynamic constants of P-curcumin association with soluble Aβ <i>(ΔH, ΔS</i>, and Δ<i>G</i>) were also determined. <b><i>Conclusion:</i></b> Cultivated curcumin weakened the direct interaction between Aβ and cell membranes and showed greater neuroprotective effects against Aβ insult than free curcumin

Supplementary Material for: Use of Epinephrine in Patients with Drug-Induced Anaphylaxis: An Analysis of the Beijing Pharmacovigilance Database

<i>Background:</i> Few studies assessing the use of epinephrine in drug-induced anaphylaxis (DIA) in the hospital setting are available. We utilized the Beijing Pharmacovigilance Database (BPD) to evaluate the appropriateness of epinephrine for DIA management. <i>Methods:</i> DIA cases collected in the BPD from January 2004 to December 2014 were adjudicated and analyzed for demographics, causative drugs, clinical signs, outcomes, initial treatment, route, dosing, and cardiovascular adverse events (CAE) of epinephrine. <i>Results:</i> DIA was primarily caused by antibiotics (38.4%), radiocontrast agents (11.9%), traditional Chinese medicine injections (10.9%), and chemotherapeutic drugs (10.3%). Only 708 (59.5%) patients received epinephrine treatment. Patients who received epinephrine were more likely to experience wheezing (<i>p</i> < 0.001) and respiratory arrest (<i>p</i> < 0.001). Among 518 patients with a complete record of the epinephrine administration route, the percentage of patients receiving it by intramuscular (IM) injection, subcutaneous (SC) injection, intravenous (IV) bolus injection, or IV continuous infusion was 16.9, 31.5, 43.5, and 8.1%, respectively. Among the 427 patients with a record of both the administration route and the dosing, an overdose was more likely with IV bolus (94.1%) in contrast to IM injection (56.6%; <i>p</i> < 0.001) or SC injection (43.7%; <i>p</i> < 0.001). Among the patients analyzed for CAE (<i>n</i> = 349), 17 patients accounted for 19 CAE, and 13 (76.5%) of these patients were overdosed with pinephrine. <i>Conclusion:</i> Underuse, inappropriate IV bolus use, and overdosing were the 3 major problems with epinephrine use in DIA in China. Educational training for health care professionals on the appropriate use of epinephrine in managing anaphylactic reactions is suggested

Supplementary Material for: Reduced ING4 Expression Is Associated with the Malignancy of Human Bladder

<b><i>Introduction:</i></b> Inhibitor of growth 4 (ING4) is a tumor suppressor. However the role of ING4 in human bladder malignancy is unknown. In this study, ING4 expression in human bladder cancer and its potential effects were studied. <b><i>Materials and Methods:</i></b> ING4 expression in 47 human bladder cancer tissues and paired adjacent normal tissues was detected by Western blotting, quantitative reverse transcription-polymerase chain reaction, and immunohistochemistry. The migration and cell cycle progression of SV-HUC-1 and T24 cells with aberrant ING4 expression were examined. <b><i>Results:</i></b> ING4 protein and mRNA were significantly decreased in bladder cancer tissues. ING4 protein level was significantly lower in the group of patients over 50 years of age. ING4 knockdown caused more rapid cell migration and increased the population of SV-HUC-1 and T24 cells in the G2-M phase. <b><i>Conclusion:</i></b> Our data suggest a close connection between aberrant ING4 expression and the carcinogenesis of human bladder cells. ING4 may be a potential target for bladder cancer chemotherapy

Supplementary Material for: Anti- Versus Pro-Inflammatory Metabololipidome Upon Cupping Treatment

<b><i>Background/Aims:</i></b> This study aimed to explore the metabololipidome in mice upon cupping treatment. <b><i>Methods:</i></b> A nude mouse model mimicking the cupping treatment in humans was established by administrating four cupping sets on the back skin for 15 minutes. UPLC-MS/ MS was performed to determine the PUFA metabolome in mice skin and blood before and after cupping treatment. The significantly changed lipids were administered in macrophages to assess the production of pro-inflammatory cytokines IL-6 and TNF-α by ELISA. <b><i>Results:</i></b> The anti-inflammatory lipids, <i>e.g</i>. PGE₁, 5,6-EET, 14,15-EET, 10S,17S-DiHDoHE, 17R-RvD1, RvD5 and 14S-HDoHE were significantly increased while pro-inflammatory lipids, <i>e.g</i>. 12-HETE and TXB₂ were deceased in the skin or plasma post cupping treatment. Cupping treatment reversed the LPS-stimulated IL-6 and TNF-α expression in mouse peritoneal exudates. Moreover, 5,6-EET, PGE₁ decreased the level of TNF-α, while 5,6-EET, 5,6-DHET downregulated IL-6 production in macrophages. Importantly, 14,15-EET and 14S-HDoHE inhibited both IL-6 and TNF-α induced by lipopolysaccharide (LPS). 17-RvD1, RvD5 and PGE₁ significantly reduced the LPS-initiated TNF-α, while TXB₂ and 12-HETE further upregulated the LPS-enhanced IL-6 and TNF-α expression in macrophages. <b><i>Conclusion:</i></b> Our results reveal the identities of anti-inflammatory <i>versus</i> pro-inflammatory metabolipidome and suggest the potential therapeutic mechanism of cupping treatment

Supplementary Material for: Association of Intravenous Tirofiban with Functional Outcomes in Acute Ischemic Stroke Patients with Acute Basilar Artery Occlusion Receiving Endovascular Thrombectomy

Introduction: The aim of this study was to test the hypothesis that intravenous tirofiban improves functional outcomes without promoting the risk of intracranial hemorrhage (ICH) in stroke secondary to basilar artery occlusion (BAO) receiving endovascular thrombectomy. Methods: Patients with acute BAO stroke who were treated with endovascular thrombectomy and had tirofiban treatment information were derived from “BASILAR”: a nationwide, prospective registry. All eligible patients were divided into tirofiban and no-tirofiban groups according to whether tirofiban was used intravenously. The primary endpoint was the 90-day severity of disability as assessed by the modified Rankin scale score. Safety outcomes were the frequency of ICH and mortality. Results: Of 645 patients included in this cohort, 363 were in the tirofiban group and 282 were in the no-tirofiban group. Thrombectomy with intravenous tirofiban reduced the 90-day disability level over the range of the modified Rankin scale (adjusted common odds ratio, 2.08; 95% confidence interval (CI), 1.45–2.97; p p p = 0.004) and symptomatic ICH (4.8% vs. 10.1%; p = 0.01) in the tirofiban group was significantly lower than that in the no-tirofiban group. Conclusions: In patients with acute BAO stroke who underwent endovascular treatment, intravenous tirofiban might be associated with favorable outcome, reduced mortality, and a decreased frequency of ICH