bluuebunny/crossref_metadata_2025 huggingface dataset:
Utility of Abdominal Drain in Gastrectomy (ADiGe) Trial: study protocol for a multicenter non-inferiority randomized trial
Authors: Weindelmayer Jacopo , Mengardo Valentina , Veltri Alessandro , Baiocchi Gian Luca , Giacopuzzi Simone , Verlato Giuseppe , de Manzoni Giovanni
Published: September 2020

Background: Prophylactic use of abdominal drain in gastrectomy has been questioned in the last 15 years and a 2015 Cochrane meta-analysis on four RCTs concluded that there was no convincing evidence to the routine drain placement in gastrectomy. Nevertheless the Authors evidenced the moderate/low quality of the included studies and highlighted how 3 out of four came from Eastern countries. After 2015 only retrospective studies have been published, all with inconsistent results.Methods: ADiGe (Abdominal Drain in GastrEctomy) Trial is a multicenter prospective randomized non-inferiority trial with a parallel design. It aimed to verify whether avoiding routine use of abdominal drain is burdened with complications, particularly an increase in postoperative invasive procedures. Patients with gastric cancer, scheduled for subtotal or total gastrectomy with curative intent, are eligible for inclusion, irrespective of previous oncological treatment. The primary composite endpoint is reoperation or percutaneous drainage procedures within 30 postoperative days. The primary analysis will verify whether the incidence of the primary composite endpoint is higher in the experimental arm, avoiding routine drain placement, than control arm, undergoing prophylactic drain placement, in order to falsify or support the null hypothesis of inferiority. Secondary endpoints assessed for superiority are overall morbidity and mortality, Comprehensive Complications Index, incidence and time for diagnosis of anastomotic and duodenal leaks, length of hospital stay, readmission rate.Assuming one-sided alpha of 5%, and cumulative incidence of the primary composite endpoint of 14.5% in control arm and 9% in the experimental one, 256 patients allow to achieve 80% power to detect a non-inferiority margin difference between the arm proportions of 4.5%. Considering a 15.5% drop-out rate, 304 patients are needed. In order to have a balanced percentage between total and subtotal gastrectomies, recruitment will end at 152 patients for each type of gastrectomy. The surgeon and the patient are blinded until the end of the operation, while postoperative course is not blinded to patient and caregivers.Discussion: ADiGe Trial could contribute to critically re-evaluate the role of prophylactic drain in gastrectomy, a still widely used procedure.Trial registration: Retrospectively registered on 01/10/2020 at Clinicaltrials.gov with the identifier NCT04227951, https://clinicaltrials.gov/ct2/show/NCT04227951.
Early Epidemiological Features and Trends of the COVID-19 Outbreak in Southeast Asia: a Population-Level Observational Study
Authors: Zhu Mingjian , Jirapat Kleepbua , Shen Jian , Chew Sien Ping , Tan Joanna Wei Hui , Latthitham Natthjija , Li Lanjuan
Published: September 2020

Background: The global outbreak of coronavirus disease 2019 (COVID-19) has been ongoing in Southeast Asia since 13 January 2020. We conducted an observational study to investigate underlying disease patterns of COVID-19 in Southeast Asia, and consequently to guide intervention strategies against the pandemic.Methods: In this population-level observational study set in Southeast Asia, we compiled a list of patients with COVID-19 (n = 925) and daily country-level case counts (n = 1346) from 13 January 2020 through 16 March 2020. All epidemiological data were extracted from official websites of the WHO and health authorities of each Southeast Asian country. Relevant spatiotemporal distributions, demographic characteristics, and short-term trends were assessed.Results: A total of 1,346 confirmed cases of COVID-19, with 217 (16.1%) recoveries and 18 (1.3%) deaths, were reported in Southeast Asia as of 16 March 2020. Early transmission dynamics were examined with an exponential regression model: y=0.30e0.13x (p<0·01, adjusted R2 = 0.96). Using this model, we predicted that the cumulative number of reported COVID-19 cases in Southeast Asia would exceed 10,000 by early April 2020. A total of 74 cities across eight countries in Southeast Asia were affected by COVID-19. Most of the confirmed cases were located in five international metropolitan areas. Demographic analyses of the 925 confirmed cases indicated a median age of 44 years and a sex ratio of 1.25. The median age of the local patient population was significantly higher than that of the corresponding country’s general population (p<0·01), whereas the sex ratio did not significantly differ.Conclusions: The COVID-19 situation in Southeast Asia is unevenly geographically distributed and pessimistic in the short term. Age may play a significant role in both the susceptibility to and outcome of infection. Real-time active surveillance and targeted intervention strategies are urgently needed to contain the pandemic.
Attitudinal and demographic factors associated with seeking help and receiving antidepressant medication for symptoms of common mental disorder
Authors: Manescu Elena Andreea , Robinson Emily J. , Henderson Claire
Published: September 2020

Background: Despite the increased attention given to improvement of mental health-related knowledge and attitudes, rates of help-seeking for mental illness remain low even in countries with well-developed mental health services. This study examines the relationships between attitudes to mental illness, symptoms of common mental disorder and seeking-help and receiving medication for a mental health problem. Methods: We used data from the nationally representative Health Survey for England 2014 to design three logistic regression models to test for the effects of attitudes to mental illness (measured by a shortened version of the Community Attitudes toward the Mentally Ill, CAMI scale) on: recent contact with a doctor for a mental health problem; use of any type of mental health service in the last 12 months; and having antidepressants currently prescribed, while controlling for symptoms of common mental disorder (measured by the General Health Questionnaire, GHQ). We also tested for an interaction between attitudes to mental illness and symptoms of common mental disorder on the outcomes. Results: A significant but very small effect of CAMI score was found on ‘antidepressants currently prescribed’ model (OR=1.01(1.00, 1.02) but not on the two indicators of help-seeking. We also found a significant but very small interaction between CAMI and GHQ scores on recent contact with a doctor (OR= 0.99, 95% CI (0.990, 0.998); adjusted Wald test P=0.01)). Knowing someone with a mental illness had a significant positive effect on help-seeking indicated by: (a) recent contact with a doctor (2.65 (1.01, 6.98)) and (b) currently prescribed antidepressant (2.67 (1.9, 3.75)) after controlling for attitudes to mental illness. Conclusions: Our results suggest that knowing someone with a mental health problem seems to have a further positive effect on help-seeking, beyond improving attitudes to mental illness. Furthermore, multiple different types and aspects of stigma may contribute to help-seeking behaviours, consequently multi-faceted approaches are likely to be most efficient.
Transcatheter closure for patent ductus arteriosus in patients with Eisenmenger syndrome: to do or not?
Authors: Xu Jing , Wang Liang , Shen Yunli , Geng Liang , Chen Fadong
Published: September 2020

Background: Patent ductus arteriosus ( PDA) complicated by Eisenmenger syndrome (ES) remains to be a major cause of morbidity and mortality worldwide. Giving increasing evidence of benefit from targeted therapies, ES patients once thought to be inoperable may have increasing options for management. This study aims to explore whether the PDA in patients with ES can be treated with transcatheter closure (TCC).Methods: Between August 2014 and July 2016, four out of fifteen PDA-ES patients whose Qp/Qs improved significantly and Qp/Qs>1.5 after acute vasodilator testing with 100% oxygen were selected to receive TCC and pulmonary vasodilator therapy. PAH-targeted drugs were prescribed before and after occlusion for all patients. Trial occlusion was performed before permanent closure.Results: The first TCC failed after the initiation of PAH-targeted drugs for 6 months in four patients. After the medication was adjusted and extended to 12 months, TCC was performed for all without hemodynamic intolerances during perioperative period. Pulmonary artery systolic pressure (PASP) was significantly decreased (≥ 40%) immediately after TCC. During a mean 52-month follow-up, there was a further decrease of PASP in two patients, the other two showed improved pulmonary vascular resistance (PVR), WHO functional class and six-minute walking distance although PASP was deteriorated again.Conclusion: Some selected PDA-ES patients with PVR<15Wood U and Qp/Qs>1.5 at baseline might benefit from TCC and PAH-targeted drugs pre- and post-occlusion play a crucial role.
Proline-rich protein 11 overexpression is associated with a more aggressive phenotype and poor overall survival in ovarian cancer patients
Authors: zhan yu , wu xueyuan , zheng gang , jin jingjing , li chaofu , yu guanzhen , li wenfeng
Published: September 2020

Background: The proline-rich protein 11 (PRR11) is a newly identified oncogene associated with a poor prognosis in several human cancers. Nonetheless, research on its role in ovarian cancer (OC) remains largely understudied. Therefore, this study aims to evaluate the expression levels of PRR11 protein and its role in human ovarian cancer. Methods: Immunohistochemistry analysis was used to evaluate the expression levels of PRR11 protein in human samples obtained from 49 patients diagnosed with OC and subjected to curative surgery in the First Affiliated Hospital of Wenzhou Medical University between 2007 and 2015. Results: In total, 57.1% of the primary OC tumor tissue evaluated demonstrated overexpression of PRR11. Meanwhile, survival analysis showed that the overall survival (OS) of patients presenting overexpression of PRR11 was significantly lower than the OS of the patients with negative PRR11. In subsequent experiments, it was found that silencing the expression of PRR11 expression inhibited the proliferation of tumor cells and the migration of cells in vitro. Further, cells subjected to PRR11 knockdown exhibited a decrease in tumor growth in vivo. The downregulation of PRR11 was coupled with a decrease in N-cadherin and downregulation in the expression of early growth response protein 1 (EGR1).Conclusions: The findings suggest that PRR11 might be considered as a potential target for prognostic assessment and gene therapy strategies for patients diagnosed with OC.
A novel analysis of the differentiated expression of long noncoding RNAs profiles in human prostate cancer
Authors: Ge Shengyang , Mi Yuanyuan , Zhao Xiaojun , Hu Qingfeng , Guo Yijun , Zhong Fan , Zhang Yang , Xia Guowei , Sun Chuanyu
Published: September 2020

Background: Long noncoding RNAs (lncRNAs) have been proved to be an important regulator in gene expression. In almost all kinds of cancers, lncRNAs participated in the process of pathogenesis, invasion, and metastasis. Meanwhile, compared with the large amounts of patients, there is rare knowledge about the role of lncRNAs in prostate cancer (PCa).Material/Method: In this study, lncRNA expression profiles of prostate cancer were detected by Agilent microarray chip, 5 pairs of case and control specimens were involved in. Differentially expressed lncRNAs were screened out by volcano plot for constructing lncRNA-miRNA-mRNA central network. Then, the top ten up-regulated and down-regulated lncRNAs were validated by qRT-PCR in another 5 tumor specimens and 7 para-cancerous/benign contrasts. Furthermore, we searched for the survival curve of the top 10 upregulated and downregulated lncRNAs.Results: A total of 817 differentially expressed lncRNAs were filtered out by the criteria of fold change (FC) and t-test p < 0.05. Among them, 422 were upregulated, whereas 395 were downregulated in PCa tissues. Gene ontology and KEGG pathway analyses showed that many lncRNAs were implicated in carcinogenesis. lnc-MYL2-4:1 (FC = 0.00141, p = 0.01909) and NR_125857 (FC = 59.27658, p = 0.00128) had the highest magnitude of change. The subsequent qPCR confirmed the expression of NR_125857 was in accordance with the clinical samples. High expression of PCA3, PCAT14 and AP001610.9 led to high hazard ratio while low expression of RP11-279F6.2 led to high hazard ratio.Conclusions: Our study detected a relatively novel complicated map of lncRNAs in PCa, which may have the potential to investigate for diagnosis, treatment and follow-up in PCa. Our study revealed the expression of NR_125857 in human PCa tissues was most up-regulated. Further studies are needed to investigate to figure out the mechanisms in PCa.
Calreticulin promotes EMT in pancreatic cancer via mediating Ca2+ dependent acute and chronic endoplasmic reticulum stress
Authors: Sheng Weiwei , Wang Guosen , Tang Jingtong , Shi Xiaoyang , Cao Rongxian , Sun Jian , Lin YiHeng , Jia Chao , Chen Chuanping , Zhou Jianping , Dong Ming
Published: September 2020

Background: Our previous study showed that calreticulin (CRT) promoted EGF-induced epithelial-mesenchymal transition (EMT) in pancreatic cancer (PC) via Integrin/EGFR-ERK/MAPK signaling. We next investigated the novel signal pathway and molecular mechanism involving the oncogenic role of CRT in PC. Methods: We investigated the potential role and mechanism of CRT in regulating intracellular free Ca2+ dependent acute and chronic endoplasmic reticulum stress (ERS)-induced EMT in PC in vitro and vivo.Results: Thapsigargin (TG) induced acute ERS via increasing intracellular free Ca2+ in PC cells, which was reversed by CRT silencing. Additionally, CRT silencing inhibited TG-induced EMT in vitro by reversing TG-induced changes of the key proteins in EMT signaling (ZO-1, E-cadherin and Slug) and ERK/MAPK signaling (pERK). TG-promoted cell invasion and migration was also rescued by CRT silencing but enhanced by IRE1α silencing (one of the key stressors in unfolded protein response). Meanwhile, CRT was co-immunoprecipitated and co-localized with IRE1α in vitro and its silencing led to the chronic ERS via upregulating IRE1α independent of IRE1-XBP1 axis. Moreover, CRT silencing inhibited IRE1α silencing-promoted EMT, including inhibiting the activation of EMT and ERK/MAPK signaling and the promotion of cell mobility. In vivo, CRT silencing decreased subcutaneous tumor size and distant liver metastasis following with the increase of IRE1α expression. A negative relationship between CRT and IRE1α was also observed in clinical PC samples, which coordinately promoted the advanced clinical stages and poor prognosis of PC patients. Conclusions: CRT promotes EMT in PC via mediating intracellular free Ca2+ dependent TG-induced acute ERS and IRE1α-mediated chronic ERS via Slug and ERK/MAPK signaling.
Transvaginal natural orifice transluminal endoscopic surgery (vNOTES) hysterectomy for uterus weighing ≥ 1kg
Authors: Wang Xiaojuan , LI Junwei , Hua Keqin , Chen Yisong
Published: September 2020

Background: The transvaginal natural orifice transluminal endoscopic surgery (vNOTES) applied in gynecology has been developed recent years and been evolving. In this study, we aimed to evaluate the feasibility and effect of the vNOTES hysterectomy for uterus ≥ 1 kilogram (kg).Methods: From January 2019 to March 2020, patients with benign indications in cases of uterus weighing ≥ 1kg, underwent vNOTES hysterectomy were studied retrospectively. The patients’ demographics, indications for surgery, operation outcomes and follow-up details were recorded. Results: 39 patients were performed vNOTES hysterectomy for large uterus (mean weight 1141.8 gram, range from 1000 to 1720), indications for surgery included bulky uterine myomas or adenomysosis. The mean age was 48 years (range 42-66) and mean BMI was 24 kg/m2 (range 18.4-38). Mean operating time was 123.3 minutes (rang 40-400) and the mean estimated blood loss was 206.7 milliliters (range 10-1300). The mean pain assessment was 2.1 (range 0-5). The mean length of stay was 2.4 nights (1-11). 1 patient experienced ureteral injury and was performed ureteral anastomosis. 3 patients were converted to vaginal-assisted trans-umbilicus single-port laparoscopy. The learning curve was analyzed to show that 20 cases were needed to achieve proficiency in vNOTES hysterectomy for large uterus ≥ 1 kg.Conclusion: Our preliminary experience suggested that vNOTES hysterectomy for large uterus weighing ≥ 1kg was feasible and safe, meanwhile this procedure had the advantages of all the minimal invasive approach such as fast recovery and aesthetic advantage.
A novel sequential treatment of palbociclib enhances the effect of cisplatin in RB-proficient triple-negative breast cancer
Authors: Huang Yajing , Wu Hao , Li Xingrui
Published: September 2020

Background: Triple-negative breast cancer (TNBC) is a highly aggressive malignancy lack of sensitivity to chemo-, endocrine and targeted therapy. CDK4/6 inhibitors, combined with endocrine therapy, have been proven to be effective in postmenopausal women with HR-positive, HER2-negative advanced or metastatic breast cancer. So we investigated that whether CDK4/6 inhibitor palbociclib (PD) could enhance effects of cisplatin (CDDP) on TNBC.Methods: The effects of different drug regimens of PD and CDDP on MDA-MB-231 and RB-knockdown MDA-MB-231 (sh-MDA-MB-231) cells were assessed in vitro and vivo. MDA-MB-468 and RB-overexpression MDA-MB-468 cells were used to assess the effect of PD-CDDP regimen in vitro. Immunoblotting illustrated cyclin D1/RB/E2F axis signaling pathway.Results: PD induced G1 phase cell cycle arrest in MDA-MB-231 cell line. However, synchronous treatment with PD and CDDP for 24h, PD used for 24h and then followed by CDDP or CDDP used for 24h and then followed by PD all had no influence on cell apoptosis of MDA-MB-231 cells. We further investigated the effect of PD or CDDP withdrawal on sequential treatment and found that PD used for 48h and then withdrawn for 48h followed by CDDP (PD-CDDP) could significantly increase apoptosis, inhibit cell viability and colony formation of MDA-MB-231 cells, while in other regimens PD and CDDP represented additive or antagonistic response. Preferential use of PD could increase DNA damage by CDDP as measured through γH2AX. These findings above were negative in sh-MDA-MB-231 cells and cell function experiments of MDA-MB-468 and RB-overexpression MDA-MB-468 cells could draw similar conclusions, which indicated that PD enhanced the sensitivity of TNBC cells to CDDP in a RB dependent manner. In vivo, this combination treatment inhibited tumor growth and Ki-67 expression compared with single drug treatments in MDA-MB-231 xenograft models. Western blotting analysis presented that PD enhanced sensitivity to CDDP through CDK4/6-cyclin D-RB-E2F pathway. Conclusions: Pre-treatment with PD synchronized tumor cell cycle through CDK4/6-cyclin D1-RB-E2F pathway, which could increase anti-tumor effect of CDDP. PD-CDDP might be an effective treatment for RB-proficient TNBC patients.
The apelin-13 peptide protects the heart against apoptosis through the ERK/MAPK and PI3K/AKT signaling pathways.
Authors: Wang Xuejun , Zhang Li , Feng Mengwen , Zhang Hao , Xu Jia , Cheng Zijie , Qian Lingmei
Published: September 2020

Background: It has been acknowledged that endocrine activity is associated with the function of multiple systems in vivo. The apelin-13 peptide has been demonstrated to play a crucial role in physiological and pathological processes. However, the function of apelin-13 peptide in doxorubicin (DOX)-induced cardiotoxicity is unknown. Methods: We explored the function and mechanism of apelin-13 peptide in apoptosis and oxidative stress by cell counting kit-8 (CCK-8) assay, trypan blue staining, TUNEL, lactate dehydrogenase (LDH), mitochondrial membrane potential assay kit with JC-1 (JC-1) and western blot in vitro. Then we verified the effect of apelin-13 in vivo by detecting serum apelin-13, CKMB, LDH, cardiac troponin I (cTnI) and cardiac troponin T (cTnT). EF, FS and LVEDs were used to identify the structural modification by echocardiography. Sirius red staining and HE staining assay were used to detecting the myocardial fibers alteration under apelin-13 treatment.Results: Treatment with apelin-13 peptide significantly enhanced cell viability, mitochondrial membrane potential, but reduced LDH release, rate of apoptotic cells and activation of caspase-3 in vitro. In mice, apelin-13 alleviated the heart dysfunction induced by DOX. 4-oxo-6-((pyrimidin-2-ylthio)methyl)-4H-pyran-3-yl 4-nitrobenzoate (ML221) inhibited the effect of extracellular signal-related kinases (ERK), phosphatidylinositol 3 kinases (PI3K) and protein kinase B (AKT) proteins phosphorylation expression compared with DOX.Conclusion: The apelin-13 and apelin receptor (APJ) interaction on the cell membrane inhibits apoptosis through the ERK/mitogen-activated protein kinase (MAPK) and PI3K/AKT signaling pathways. Our research gives a first glimpse on the biological function and mechanism of apelin-13 on cardiotoxicity.
Pediatric Tuina for the Treatment of Attention Deficit Hyperactivity Disorder (ADHD) Symptoms in Preschool Children: Study Protocol for a Pilot Randomized Controlled Trial
Authors: Chen Shu Cheng , Yu Juan , Suen Lorna Kwai-Ping , Sun Yan , Pang Ya Zheng , Wang Dong Dong , Zhao Wen Xia , Yeung Jerry Wing Fai
Published: September 2020

Background: Medication and behavior therapy are the conventional treatments for attention deficit hyperactivity disorder (ADHD), but they have limitations for preschool children. Evidence suggests that pediatric tuina, which is a modality of traditional Chinese medicine, might have beneficial effects on this condition. Therefore, we aim to preliminarily assess the effects and feasibility of parent-administered pediatric tuina for ADHD symptoms in preschoolers.Methods: It is a single-center, two-arm, parallel, open-label, pilot randomized controlled trial. Sixty children with pre-specified ADHD symptoms (hyperactivity, anxiety, and sleep disturbance) together with one of their parents will be recruited and randomized into two groups at a 1: 1 ratio. Parents in the parent-administered tuina group (intervention group, n=30) will attend an online training program to learn pediatric tuina skills for ADHD symptoms and conduct this treatment on their children at home. Parents in the parent-child interaction group (comparison group, n=30) will attend an online training about progressive muscle relaxation exercise and do this exercise with their children at home. Additional teaching materials will be provided to the participants in both groups. Both interventions should be carried out every other day during a 2-month treatment period, with each time around 20 minutes. Assessment will be performed at baseline, week4, and week 8. The primary outcome measure is the Swanson, Nolan and Pelham Parent Scale; the secondary outcomes include Preschool Anxiety Scale, Children's Sleep Habits Questionnaire, and Parental Stress Scale. A process evaluation embedded within the outcome evaluation will be performed. Differences in the scale scores and test parameters between groups will be examined using a linear mixed-effects model. Qualitative data will be analyzed using thematic content analysis, facilitated by QSR NVivo.Discussion: This study will provide evidence on the acceptability and feasibility of pediatric tuina for ADHD in preschool children. The process evaluation will help to better understand the facilitators and barriers of the intervention functioning.Trial registration: The study was registered at ClinicalTrials.gov (Identifier: NCT04237259) on 14 February 2020. Protocol version: 2, date June 23, 2020
Splenic Infarction and Infectious Diseases in Korea
Authors: Im Jae Hyoung , Chung Moon-Hyun , Lee Hye-Jin , Kwon Hea Yoon , Baek JiHyeon , Jang Ji-Hun , Lee Jin-Soo
Published: September 2020

BackgroundThe spleen contains immune cells and exhibits a pattern of infarction different from other organs; as such, splenic infarction (SI) may provide important clues to infection. However, the nature of the relationship between SI and infectious disease(s) is not well understood. Accordingly, this retrospective study investigated the relationship between SI and infection. MethodsHospital records of patients with SI, who visited Inha University Hospital (Incheon, Republic of Korea) between January 2008 and December 2018, were reviewed. Patient data regarding clinical presentation, causative pathogens, risk factors, and radiological findings were collected and analyzed.ResultsOf 353 patients with SI, 101 with infectious conditions were enrolled in this study, and their data were analyzed to identify associations between SI and infection. Ten patients were diagnosed with infective endocarditis (IE), and 26 exhibited bacteremia without IE. Twenty-seven patients experienced systemic infection due to miscellaneous causes (negative result on conventional automated blood culture), including the following intracellular organisms: parasites (malaria [n =12], babesiosis [n =1]); bacteria (scrub typhus [n =5]); viruses (Epstein–Barr [n =1], cytomegalovirus [n =1]); and unidentified pathogen[s] (n =7). Splenomegaly was more common among patients with miscellaneous systemic infection; infarction involving other organs was rare. Thirty-eight patients had localized infections (e.g., respiratory, intra-abdominal, or skin and soft tissue infection), and most (35 of 38) had other risk factors for SI. ConclusionsIn this study, various infectious conditions were found to be associated with SI, and intracellular organisms were the most common causative pathogens. Further studies are needed to examine other possible etiologies and the underlying pathophysiological mechanisms.
Melphalan induces cardiotoxicity through oxidative stress in cardiomyocytes derived from human induced pluripotent stem cells
Authors: Liu Rui , Li Dong , Sun Fangxu , Rampoldi Antonio , Maxwell Joshua , Wu Ronghu , Fischbach Peter , Castellino Sharon , Du Yuhong , Fu Haian , Mandawat Anant , Xu Chunhui
Published: September 2020

Background. Treatment-induced cardiotoxicity is a leading noncancer-related cause of acute and late onset morbidity and mortality in cancer patients on antineoplastic drugs such as melphalan—increasing clinical case reports have documented that it could induce cardiotoxicity including severe arrhythmias and heart failure. As the mechanism by which melphalan impairs cardiac cells remains poorly understood, here we aimed to use cardiomyocytes derived from human induced pluripotent stem cells (hiPSC-CMs) to investigate the cellular and molecular mechanisms of melphalan-induced cardiotoxicity and to explore potential targeted therapeutics.Methods. hiPSC-CMs were generated and treated with clinically relevant doses of melphalan. In order to characterize melphalan-induced cardiotoxicity, cell viability and apoptosis were quantified at various treatment durations. Ca2+ transient and contractility analyses were used to examine the alterations of hiPSC-CM function. Proteomic analysis, reactive oxygen species detection, and RNA-Sequencing were conducted to investigate underlying mechanisms. Results. Melphalan treatment of hiPSC-CMs induced oxidative stress, caused Ca2+-handling defects and dysfunctional contractility, altered global transcriptomic and proteomic profiles, and resulted in apoptosis and cell death. The antioxidant N-acetyl-L-cysteine attenuated these genomic, cellular and functional alterations. In addition, several other signaling pathways including the p53 and transforming growth factor-β signaling pathways were also implicated in melphalan-induced cardiotoxicity according to the proteomic and transcriptomic analyses.Conclusions. Melphalan induces cardiotoxicity through the oxidative stress pathway. This study provides a unique resource of the global transcriptomic and proteomic datasets for melphalan-induced cardiotoxicity and can potentially open up new clinical mechanism-based targets to prevent and treat melphalan-induced cardiotoxicity.
Increasing use of linezolid in a tertiary NICU during a 10-year period: reasons and concerns for the future
Authors: Matrat Lucie , Plaisant Frank , Barreto Christine , Claris Olivier , Butin Marine
Published: September 2020

Background: Linezolid has been increasingly used in tertiary NICUs. The objectives of this study were to explore the indications of these linezolid prescriptions, to analyze a possible misuse and to provide solutions to avoid such misuse.Methods: A monocentric retrospective cohort study included all neonates hospitalized in one tertiary NICU between January 1st, 2010 and December 31st, 2019 and who received at least one administration of linezolid. These data were confronted to epidemiological and antibiotic use data from the same NICU. Two independent pediatricians secondarily classified linezolid uses as adequate or not.Results: During the study period, 66 infections in 57 patients led to linezolid use. Most patients were pre-term and 21 patients (37%) died. Infections were mainly related to methicillin-resistant coagulase negative staphylococci and were frequently either pneumoniae (35%) or isolated bacteremia (48%), including 25 persistent bacteremia (64% of the 39 bacteremia). Need for a better tissue distribution or first-line treatment failure were the main reasons to initiate linezolid. Linezolid was administered for a median duration of 7 [3;10] days. No side effects were reported. Twenty-two (33%) of the 66 linezolid prescriptions were retrospectively classified as inadequate. Conclusions: A rapid increase in linezolid prescriptions has been observed in our tertiary NICU, from 2014 to 2019, with 33% inadequate uses. This worrisome trend should lead to search for therapeutic alternatives and to work on antibiotic stewardship to prevent the emergence of new antimicrobial bacterial resistance.
Navigate: A study protocol for a randomized controlled trial of an online treatment decision aid for men with low risk prostate cancer and their partners
Authors: Schofield Penelope , Gough Karla , Hyatt Amelia , White Alan , Frydenberg Mark , Chambers Suzanne , Gordon Louisa , Gardiner Robert , Murphy Declan , Cavedon Lawrence , Richards Natalie , Murphy Barbara , Quinn Stephen , Juraskova Ilona
Published: September 2020

Background
Active surveillance (AS) is the disease management option of choice for low risk prostate cancer. Despite this, men with low risk prostate cancer (LRPC) find management decisions distressing and confusing. We developed Navigate, an online decision aid to help men and their partners make management decisions consistent with their values. The aims are to evaluate the impact of Navigate on: uptake of AS; decision-making preparedness; decisional conflict, regret and satisfaction; quality of illness communication; and prostate cancer-specific quality of life and anxiety. In addition, the healthcare cost impact, cost-effectiveness and patterns of use of Navigate will be assessed. This paper describes the study protocol.
Methods
304 men and their partners are randomly assigned one-to-one to Navigate or to the control arm. Randomisation is electronically generated and stratified by site. Navigate is an online decision aid that presents up-to-date, unbiased information on LRPC tailored to Australian men and their partners including each management option and potential side-effects; and an interactive values clarification exercise. Participants in the control arm will be directed to the website of Australia’s peak national body for prostate cancer. Eligible patients will be men within three months of being diagnosed with LRPC, aged 18 years or older and who are yet to make a treatment decision and who are deemed eligible for AS by their treating clinician, and who have internet access and sufficient English to participate. The primary outcome is self-reported uptake of AS as the first-line management option. Secondary outcomes include self-reported preparedness for decision-making; decisional conflict, regret and satisfaction; quality of illness communication; and prostate cancer-specific quality of life. Uptake of AS one month after consent will be determined through patient self-report. Men and their partners will complete study outcome measures before randomisation, and one, three and six months after study consent.
Discussion
The Navigate online decision aid has the potential to increase the choice of AS in LRPC, avoiding or delaying unnecessary radical treatments and associated side effects. In addition, Navigate is likely to reduce patients’ and partners’ confusion and distress in management decision-making, and increase their quality of life.
Incidence and predictors of diabetic ketoacidosis among children with diabetes in west and east gojjam zone referral hospitals north west Ethiopia, 2019.
Authors: assefa birtukan , Zeleke Haymanot , Murugan Rajalakshmi , Wondwossen Kalkidan
Published: September 2020

Background: Recurrent diabetic ketoacidosis in patients with already diagnosed diabetic mellitus remains a relevant problem in pediatric with an incidence of 1–10% per patient. Children may die due to cerebral edema and had significant mortality (24%) and morbidity (35%).Objective: We assess the incidence and predictors of diabetic ketoacidosis among diabetic children at East and West Gojjam zone referral hospitals, North West Ethiopia, 2019.Methods: An institution-based retrospective follow up study was conducted on children who were registered from January 1, 2014, to January 1, 2019. Epi data version 3.1 & Stata 14 were used for data entering and analysis respectively.Result: A total of 354 children were included from this 207(58.5%) had diabetic ketoacidosis. The overall incidence rate of diabetic ketoacidosis was 2.27/100 children/month of observation. Age <5 years (AHR: 3.52, 95% CI (2.25, 5.49), non-adherence (AHR: 1.54, 95% CI (1.11, 2.14), inappropriate insulin storage (AHR: 1.36, 95% CI (1.008, 1.85), presence of upper respiratory tract infections during diabetic ketoacidosis diagnose (AHR: 2.22, 95% CI (1.11, 4.45) and preceding gastroenteritis (AHR: 2.18, 95% CI (1.07, 4.44) were significant predictors. Conclusion: age <5 years old, non-adherence, inappropriate insulin placement at home, Preceding gastroenteritis, and presence of upper respiratory tract infections at the time of diabetic ketoacidosis development were significant predictors. Hence, assessing and close monitoring as well as strengthened diabetic education should be given for the above predictors.
Neuromedin B Promotes Chondrocyte Differentiation of Mesenchymal Stromal Cells via Calcineurin and Calcium Signaling
Authors: Maumus Marie , Fonteneau Guillaume , Ruiz Maxime , Assou Said , Boukhaddaoui Hassan , Pastoureau Philippe , De Ceuninck Frédéric , Jorgensen Christian , Noël Danièle
Published: September 2020

Background: Articular cartilage is a complex tissue with poor healing capacities. Current approaches for cartilage repair based on mesenchymal stromal cells (MSCs) are often disappointing because of the lack of relevant differentiation factors that could drive MSC differentiation towards a stable mature chondrocyte phenotype. Methods: We used a large-scale transcriptomic approach to identify genes that are modulated at early stages of chondrogenic differentiation using the reference cartilage micropellet model. Results: We identified several modulated genes and selected neuromedin B (NMB) as one of the early and transiently modulated genes. We found that the timely regulated increase of NMB was specific for chondrogenesis and not observed during osteogenesis or adipogenesis. Furthermore, NMB expression levels correlated with the differentiation capacity of MSCs and its inhibition resulted in impaired chondrogenic differentiation indicating that NMB is required for chondrogenesis. We further showed that NMB activated the calcineurin activity through a Ca++-dependent signaling pathway. Conclusion: NMB is a newly described chondroinductive bioactive factor that upregulates the key chondrogenic transcription factor Sox9 through the modulation of Ca2+ signaling pathway and calcineurin activity.
Quantifying nanodiamonds biodistribution in whole cells with correlative iono-nanoscopy
Authors: Mi Zhaohong , Tan Hong Qi , Chen Ce-Belle , Dou Yanxin , Yang Chengyuan , Turaga Shuvan Prashant , Watt Frank , Bettiol Andrew
Published: September 2020

Correlative imaging and quantification of intracellular nanoparticles with the underlying ultrastructure is crucial for understanding cell-nanoparticle interactions in biological research. However, correlative nanoscale imaging of whole cells still remains a daunting challenge. Here, we report a straightforward nanoscopic approach for whole-cell correlative imaging, by simultaneous ionoluminescence and ultrastructure mapping implemented with a highly focused beam of alpha particles. We demonstrate that fluorescent nanodiamonds exhibit fast, ultrabright and stable emission upon excitation by alpha particles. Thus, by using fluorescent nanodiamonds as imaging probes, our approach enables quantification and correlative localization of single nanodiamonds within a whole cell at sub-40 nm resolution. As an application example, we show that our approach, together with Monte Carlo simulations, can be employed to provide unique insights into the mechanisms of nanodiamond radiosensitization at the single whole-cell level. These findings may benefit clinical studies of radio-enhancement effects by nanoparticles in charged-particle cancer therapy.
Influence of Spin Finish on Degradation, Functionalization and Long-term Storage of Polyethylene Terephthalate Fabrics Dedicated to Ligament Prostheses
Authors: Nguyen Tuan Ngoc , Rangel Andre , Grainger David W. , Migonney Véronique
Published: September 2020

Spin finish oil applied to poly(ethylene terephthalate) (PET) fibers is shown to alter the surface properties of commercial PET fibers in storage over extended storage times. Oil removal by solvent extraction as required for their applications is shown to be changed; fiber surface chemistry, particularly surface functionalization with anionic polymer grafts, is altered, and surface mechanical properties are altered. Spin finish oxidation in storage is proposed to produce these fiber changes in storage important to their biomedical performance. Background: Poly(ethylene terephthalate) (PET) fabrics surface-functionalized using anionic polymer grafts to enhance their biocompatibility, cell adhesion, proliferation and functional performance as PET ligament prostheses have been developed for medical application in vascular and ligament prostheses. Here, we provide new evidence for deleterious effects of uncontrolled storage times and conditions on the final properties of PET medical fabrics and devices, specifically alteration and degradation of applied spin finish oil and fabric fiber surface properties, and limits to surface functionalization of PET fibers important to for medical uses.Results: Textile spin finish oil effects from 2- to 25-year storage times on PET fiber degradation and surface functionalization with anionic polymers were analyzed using FTIR, DSC and by quantitative AFM nano-mechanical profiling. Degradation of the spin-finish oil/fiber interface reduced oil Soxhlet extraction efficiency due to oil solubility changes in diethyl ether or n-hexane extraction solvents. However, solvent tetrahydrofuran was shown to be the most efficient extraction solvent even after long fabric storage times, facilitating further efficient surface functionalization of PET fabrics. Surface mechanical properties of PET fibers and fabrics over storage times spanning 2 to 25 years were investigated by using AFM-Peakforce QNM. Results showed significant and dramatic reduction of the surface elastic modulus of degraded PET fiber surfaces, with surface stiffness decreasing from 2.3 GPa for optimal (2-year) conditions of storage (PET 2018) to 50-85 MPa for extended storage (to 25 year) periods (PET2009 and PET1993). Conclusion: The ambient aging of textile spin finish oil with PET surfaces was shown to induce PET surface degradation, limiting oil removal, limiting further PET fiber surface graft functionalization, and compromising mechanical properties. Moreover, residual degraded finishing oils likely contained oxidation products from extended storage that alter PET fabrics.
Highly Efficient Production of an Influenza H9N2 Vaccine Using MDCK Suspension Cells
Authors: Wu Yixiao , Jia Hanjing , Lai Hanzhang , Liu Xuping , Tan Wen-Song
Published: September 2020

H9N2 subtype avian influenza virus poses a constant threat to the poultry industry and the control of the disease leans upon the use of effective vaccines. As an alternative to the conventional chicken embryonated eggs, animal cell culture could overcome the limitations of egg supplies and upgrade the manufacturing of avian influenza vaccines for poultry. Development of serum-free suspension cell culture could allow even higher virus productivity, where a suspension cell line with good growth and production performance is required. In this work, an adherent MDCK cell line was adapted to suspension growth to cell concentration up to 12 × 106 cells/mL in a serum-free medium in batch cultures. Subsequently, the influenza virus propagation in this MDCK cell line was evaluated and was improved with the medium exchange at time of infection as well as optimization of infection conditions in terms of MOI and cell concentration for infection. Furthermore, various feed strategies were tested in the infection phase for improved virus titer and a maximum hemagglutinin titer of 13 log2 (HAU/50 μL) was obtained using the 1:2 medium dilution strategy. Evaluation of MDCK cell growth and H9N2 virus propagation in the bioreactors with optimized operating conditions showed comparable cell performance and virus yield compared to shake flasks, with a high cell-specific virus yield above 14000 virions/cell. With the purified H9N2 virus harvested from the bioreactor, the MDCK cell-derived vaccine was able to induce high titers of neutralizing antibodies in chickens. Overall, the results demonstrate the promising application of the highly efficient MDCK cell-based production platform for the avian influenza vaccine manufacturing.
Development and Evaluation of Two-Parameter Linear Free Energy Models for the Prediction of Human Skin Permeability Coefficient of Neutral Organic Chemicals
Authors: Naseem Sana , Zushi Yasuyuki , Nabi Deedar
Published: September 2020

The experimental values of skin permeability coefficients, required for dermal exposure assessment, are not readily available for many chemicals. The existing estimation approaches are either less accurate or require many parameters that are not readily available. Furthermore, current estimation methods are not easy to apply to complex environmental mixtures. We present two models to estimate the skin permeability coefficients of neutral organic chemicals. The first model, referred to here as the 2-parameter partitioning model (PPM), exploits a linear free energy relationship (LFER) of skin permeability coefficient with a linear combination of partition coefficients for octanol-water and air-water systems. The second model is based on the retention time information of nonpolar analytes on comprehensive two-dimensional gas chromatography (GC×GC). The PPM successfully explained variability in the skin permeability data (n = 175) with R2 = 0.82 and root mean square error (RMSE) = 0.47 log unit. In comparison, the US-EPA’s model DERMWIN exhibited an RMSE of 0.78 log unit. The Zhang model T a 5-parameter LFER equation based on experimental Abraham solute descriptors (ASDs) h performed slightly better with an RMSE value of 0.44 log unit. However, the Zhang model is limited by the scarcity of experimental ASDs. The GC×GC model successfully explained the variance in skin permeability data of nonpolar chemicals (n = 79) with R2 = 0.90 and RMSE = 0.23 log unit. The PPM can easily be implemented in US-EPA’s Estimation Program Interface Suite (EPI Suite™). The GC×GC model can be applied to the complex mixtures of nonpolar chemicals.
Inhibition of BACE1 Facilitates Macrophage-based Immunotherapy to Suppress Malignant Growth of Glioblastoma
Authors: Zhai Kui , Huang Zhi , Tao Weiwei , Fang Xiaoguang , Huang Qian , Li Xiaoxia , Stark George , Hamilton Thomas , Bao Shideng
Published: September 2020

Malignant tumors, including glioblastoma (GBM), contain abundant tumor-associated macrophages (TAMs) that mainly promote tumor growth and therapeutic resistance. Reprograming tumor-promoting TAMs (pTAMs) into tumor-suppressive TAMs (sTAMs) represents an attractive therapeutic strategy. We discovered that inhibition of the β-site amyloid precursor protein cleaving enzyme 1 (BACE1) by MK-8931 potently redirects pTAMs into sTAMs and promotes macrophage phagocytosis of glioma cells to suppress the malignant growth of GBM. Moreover, low doses of radiation markedly enhance TAM infiltration and synergize with MK-8931 treatment. BACE1 is preferentially expressed by pTAMs in human GBMs and is required for maintaining pTAM polarization through trans-IL-6/sIL-6R/STAT3 signaling. As several BACE1 inhibitors, including MK-8931, previously developed for Alzheimer's disease, were shown in clinical trials to be safe for humans, repurposing these inhibitors for cancer therapy should be straightforward. Collectively, this study offers a promising therapeutic approach, through inhibition of BACE1, to facilitate the macrophage-based tumor immunotherapy.
Mesenchymal Stem Cells-Secreted TGF-β1 Restores Treg/Th17 Skewing Induced by Lipopolysaccharide and Hypoxia Challenge via miR-155 Suppression
Authors: Xue Ming , Zhang Xiwen , Liu Feng , Chen Jianxiao , Xu Jingyuan , Xie Jianfeng , Yang Yi , Yu Weiping , Qiu Haibo
Published: September 2020

Background: Regulatory T cells (Treg)/T helper (Th) 17 skewing plays a crucial role in development of acute respiratory distress (ARDS). Mesenchymal stem cells (MSCs)-secreted transforming growth factor (TGF)-β1 has remarkable immunomodulatory effects on CD4+T cells, being environment sensitive and remains lack of discussion in hypoxic and inflammatory condition of ARDS.Methods: Purified mice splenic CD4+ T cells were pre-coated with anti-CD3 (5ug/ml) and anti-CD28 (2ug/ml) overnight. RAW264.7 cells were added as antigen presenting cells (APCs). T cells with and without RAW264.7 cells were treated with various LPS concentrations of 0,10,100,1000ng/ml or/and at hypoxia condition of 5% O2. Based on LPS (100ng/ml) and hypoxia condition (5% O2) as stimuli, MSCs were set in direct coculture or indirect coculture methods by transwell system. Anti-TGF-β1 neutralization antibody was added to explore the role of TGF-β1 among the soluble factors secreted by MSCs; miR-155 overexpression of CD4+T cells were performed by transfection and then were added to the MSCs-CD4+T cells coculture system in hypoxic and LPS-stimulated condition. After 48 hours, cells or supernatant were collected for detection of frequency of Treg and Th17 subsets, CD4+T cells apoptosis and proliferation capacity assay by flowcytometry, secretions of INF-γ, IL-17A, IL-21, TGF-β1 and IL-10 by ELISA, levels of miR-155, Rorc, Foxp3 and Ptpn2 mRNA expression of CD4+T cells by RT-PCR.Results: MSCs could restore the skewed Treg/Th17 induced by LPS and hypoxia as compared to groups without MSCs with increased secretion of TGF-β1, IL-10 and IL-17A(P<0.05) and attenuate the increased expression of miR-155 in CD4+T cells, which was independent on cell-to-cell contact mechanism while TGF-β1 neutralization could significantly inhibit the effects of MSCs restoring the skewed Treg/Th17 and abolished its effect on miR-155 expression in CD4+T cells.Conclusions: These findings suggested miR-155 suppression of CD4+T cells mediated MSCs-secreted TGF-β1 modulating the skewed Treg/Th17 induced by LPS-hypoxia challenge, providing evidence when proposing future T lymphocyte-targeted cell therapy in ARDS.