bluuebunny/crossref_metadata_2025_split huggingface dataset:
Development and evaluation of recombinant GRA8 protein for the serodiagnosis of Toxoplasma gondii infection in goats
Authors: Jirapattharasate Charoonluk , Udonsom Ruenruetai , Prachasuphap Apichai , Jongpitisub Kodcharad , Dhepakson Panadda 
Published: October 2020

Abstract
BackgroundThe development of sensitive and specific methods for detecting Toxoplasma gondii infection is critical for preventing and controlling toxoplasmosis in humans and other animals. Recently, various recombinant proteins have been used in serological tests for diagnosing toxoplasmosis. The production of these antigens is associated with live tachyzoites obtained from cell cultures or laboratory animals for genomic extraction to amplify target genes. Synthetic genes have gained a key role in recombinant protein production. For the first time, we demonstrated the production of the recombinant protein of the T. gondii dense granular antigen 8 (TgGRA8) gene based on commercial gene synthesis. Recombinant TgGRA8 plasmids were successfully expressed in an Escherichia coli system. The recombinant protein was affinity-purified and characterized via sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. Furthermore, the diagnostic potential of the recombinant protein was assessed using 306 field serum samples from goats via indirect enzyme-linked immunosorbent assay (iELISA) and the latex agglutination test (LAT).ResultsWestern blotting using known positive serum samples from goats identified a single antigen at the expected molecular weight of TgGRA8 (27 kDa). iELISA illustrated that 15.40% of goat samples were positive for T. gondii-specific IgG antibodies. In addition, TgGRA8 provided high sensitivity and specificity, with significant concordance (91.83) and kappa values (0.69) compared with the results obtained using LAT.ConclusionOur findings highlight the production of a recombinant protein from a synthetic TgGRA8 gene and the ability to detect T. gondii infection in field samples. The sensitivity and specificity of TgGRA8 demonstrated that this protein could be a good serological marker for detecting specific IgG in goat sera
CircRNA PIP5K1A promotes the progression of glioma through upregulation of TCF12/PI3K/AKT pathway by sponging miR-515-5p
Authors: Zheng Kebin , Xie Haipeng , Wu Xiaosong , Wen Xichao , Zeng Zhaomu , Shi Yanfang 
Published: October 2020

Abstract
BackgroundIncreasing studies have revealed that circular RNAs (CircRNAs) make great contribution to regulating tumor progression. Therefore, we intended to explore the expression characteristics, function, and related mechanisms of a novel type of circRNA, PIP5K1A in glioma. MethodsFirstly, RT-PCR was carried out to examine CircPIP5K1A expression in glioma tissues and adjacent normal tissues, and the correlation between CircPIP5K1A level and the clinical pathological indicators of glioma was analyzed. Then, the CircPIP5K1A expression in various glioma cell lines was detected, and a cell model of CircPIP5K1A overexpression and knockdown was constructed. Subsequently, cell proliferation and viability were detected by CCK8 method and BrdU staining, apoptosis was detected by flow cytometry, and cell invasion was examined by Transwell assay. The expression of TCF12, PI3K/AKT pathway apoptotic related proteins (including Caspase3, Bax and Bcl2) and epithelial-mesenchymal transition (EMT) markers (including E-cadherin, Vimentin and N-cadherin) by western blot or RT-PCR. ResultsThe results manifested that CircPIP5K1A was obviously upregulated in glioma tissues (compared with that in normal adjacent tissues), and overexpressed CircPIP5K1A was distinctly related to glioma volume and histopathological grade. Functionally, overexpressing CircPIP5K1A notably elevated the proliferation, invasion, EMT of glioma cells, and inhibited apoptosis both in vivo and in vitro. Besides, CircPIP5K1A also upregulated TCF12 and PI3K/AKT pathway activation. Bioinformatics analysis testified that miR-515-5p was a common target of CircPIP5K1A and TCF12, while dual luciferase reporter assay and RNA immunocoprecipitation (RIP) experiment further confirmed that CircPIP5K1A targeted miR-515-5p, which bound the 3'-untranslated region (UTR) of TCF12. ConclusionsAltogether, the study illustrated that CircPIP5K1A is a potential prognostic marker in glioma and regulates the development of glioma through the modulating miR-515-5p mediated TCF12/PI3K/AKT axis.
From SARS-CoV-2 hematogenous spreading to endothelial dysfunction: clinical-histopathological study of cutaneous signs of COVID-19.
Authors: Patrì Angela , Vargas Maria , Buonanno Pasquale , Annunziata Maria Carmela , Russo Daniela , Staibano Stefania , Servillo Giuseppe , Fabbrocini Gabriella 
Published: October 2020

Abstract
Background: To date, very few studies on clinical-histopathological correlations of cutaneous disorders associated with COVID-19 have been conducted. Case presentation: The Case 1 was a 90-year-old man, who tested positive for SARS-CoV-2 from a nasopharyngeal swab. Two days later, he was hospitalized and after eleven days transferred to Intensive Care Unit. A chest CT showed bilateral ground-glass opacities. Just that day, an erythematous maculo-papular rash appeared on trunk, shoulders and neck, becoming purpuric after few days. Histological evaluations revealed a chronic superficial dermatitis with purpuric aspects. The superficial and papillary dermis appeared edematous, with a perivascular lympho-granulocytic infiltrate and erythrocytic extravasation. At intraepithelial level, spongiosis and a granulocyte infiltrate were detected. Arterioles, capillaries and post-capillary venules showed endothelial swelling and appeared ectatic. The patient was treated with hydroxychloroquine, azithromycin, lopinavir-ritonavir and tocilizumab. Regrettably, due to severe lung impairment, he died.The Case 2 was a 85-year-old man, admitted to Intensive Care Unit, where he was intubated. He had tested positive for SARS-CoV-2 from a nasopharyngeal swab two days before. A chest RX showed bilateral atypical pneumonia. After seven days, a cutaneous reddening involving trunk, upper limbs, neck and face developed, configuring a sub-erythroderma. Histological evaluations displayed edema in the papillary and superficial reticular dermis, and a perivascular lymphocytic infiltrate in the superficial dermis. The patient was treated with hydroxychloroquine, azithromycin, lopinavir-ritonavir and tocilizumab. Sub-erythroderma as well as respiratory symptoms gradually improved until healing. Conclusions: The endothelial swelling detected in the Case 1 could be a morphological expression of SARS-CoV-2-induced endothelial dysfunction. We hypothesize that cutaneous damage could be initiated by endothelial dysfunction, caused by SARS-CoV-2 infection of endothelial cells or induced by immune system activation. The disruption of endothelial integrity could enhance microvascular permeability, extravasation of inflammatory cells and cytokines, with cutaneous injury. The Case 2 developed a sub-erythroderma associated with COVID-19, and a non-specific chronic dermatitis was detected at histological level. We speculate that a purpuric rash could represent the cutaneous sign of a more severe coagulopathy, as highlighted histologically by vascular abnormalities, while a sub-erythroderma could be expression of viral hematogenous spreading, inducing a non-specific chronic dermatitis.
A novel circular RNA hsa_circRNA_103809/miR-377-3p/GOT1 pathway regulates cisplatin-resistance in non-small cell lung cancer (NSCLC)
Authors: Zhu Xiang , Han Jing , Lan Huiyin , Lin Qingren , Wang Yuezhen , Sun Xiaojiang 
Published: October 2020

Abstract
Background
Cisplatin is the first-line chemotherapeutic drug for non-small cell lung cancer (NSCLC), and emerging evidences suggests that targeting circular RNAs (circRNAs) is an effective strategy to increase cisplatin-sensitivity in NSCLC, but the detailed mechanisms are still not fully delineated.
Methods
Cell proliferation, viability and apoptosis were examined by using the cell counting kit-8 (CCK-8) assay, trypan blue staining assay and Annexin V-FITC/PI double staining assay, respectively. The expression levels of cancer associated genes were measured by using the Real-Time qPCR and Western Blot analysis at transcriptional and translated levels. Dual-luciferase reporter gene system assay was conducted to validated the targeting sites among hsa_circRNA_103809, miR-377-3p and 3’ untranslated region (3’UTR) of GOT1 mRNA. The expression status, including expression levels and localization, were determined by immunohistochemistry (IHC) assay in mice tumor tissues.
Results
Here we identified a novel hsa_circRNA_103809/miR-377-3p/GOT1 signaling cascade contributes to cisplatin-resistance in NSCLC in vitro and in vivo. Mechanistically, parental cisplatin-sensitive NSCLC (CS-NSCLC) cells were subjected to continuous low-dose cisplatin treatment to generate cisplatin-resistant NSCLC (CR-NSCLC) cells, and we found that hsa_circRNA_103809 and GOT1 were upregulated, while miR-377-3p was downregulated in CR-NSCLC cells but not in CS-NSCLC cells. In addition, hsa_circRNA_103809 sponged miR-337-3p to upregulate GOT1 in CS-NSCLC cells, and knock-down of hsa_circRNA_103809 enhanced the inhibiting effects of cisplatin on cell proliferation and viability, and induced cell apoptosis in CR-NSCLC cells, which were reversed by downregulating miR-377-3p and overexpressing GOT1. Consistently, overexpression of hsa_circRNA_103809 increased cisplatin-resistance in CS-NSCLC cells by regulating the miR-377-3p/GOT1 axis. Finally, silencing of hsa_circRNA_103809 aggravated the inhibiting effects of cisplatin treatment on NSCLC cell growth in vivo.
Conclusions
Analysis of data suggested that targeting the hsa_circRNA_103809/miR-377-3p/GOT1 pathway increased susceptibility of CR-NSCLC cells to cisplatin, and this study provided novel targets to improve the therapeutic efficacy of cisplatin for NSCLC treatment in clinic.
Thyroid function and non-alcoholic fatty liver disease in hyperthyroidism patients
Authors: Wang Bairong , Wang Baomin , Yang Yumei , Xu Jing , Hong Mengyang , Xia Mingfeng , Li Xiaomu , Gao Xin 
Published: October 2020

Abstract
Background: Although thyroid function has been demonstrated to be associated with non-alcoholic fatty liver disease (NAFLD) in different population, the prevalence and features of NAFLD in hyperthyroidism have not been reported. The present study aims to investigate the prevalence of NAFLD and association of thyroid function and NAFLD in hyperthyroidism patients. Methods: This cross-sectional study was performed in Zhongshan Hospital, Fudan University, China. A total 117 patients with hyperthyroidism were consecutively recruited from 2014-2015. Thyroid function and other clinical features were measured, liver fat content was measured by color Doppler ultrasonically, NFALD NAFLD was defined in patients with liver fat content more than 9.15% . Statistical analyses were performed with SPSS software package version 13.0.Results: The prevalence of NAFLD was 11.97% in hyperthyroidism. Patient with NAFLD had lower free triiodothyronine (FT3)fT3 and free thyroxine (FT4)fT4 levels than patients without NAFLD (P < 0.05). After adjusting for age, gender, metabolic parameters and inflammation factors, higher fT3 FT3 were associated with lower liver fat content (β=－0.072, P=0.009) and decreased odds ratio of NAFLD (OR=0.267, 95%CI 0.087-0.817, P=0.021). Conclusions: The prevalence of NAFLD was 11.97% in hyperthyroidism patients, and FT3 level was negatively associated with the liver fat content in this population. These results may provide new evidence in the role of thyroid hormone on the regulation of liver fat content and NAFLD.
“Can Virtue Be Taught?”: A Content Analysis of Medical Students’ Opinions of the Professional and Ethical Challenges to their Professional Identity Formation
Authors: Hawking Michael , Kim Jenny , Jih Melody , Hu Chelsea , Yoon John D. 
Published: October 2020

Abstract
Background: Efforts have begun to characterize the ethical and professional issues encountered by medical students in their clinical years. By applying previously identified taxonomies to a national sample of medical students, this study seeks to develop generalizable insights that can inform professional identity formation across various clerkships and medical institutions.Methods: In a national survey of medical students, participants answered an open-ended survey item that asked them to describe a clinical experience involving an ethical or professional issue. We conducted a content analysis with these responses using the Kaldjian taxonomy of ethical and professionalism themes in medical education through an iterative, consensus-building process. Noting the emerging virtues-based approach to ethics and professionalism, we also reexamined the data using a taxonomy of virtues.Results: The response rate to this survey item was 144 out of 499 eligible respondents (28.9%). All 144 responses were successfully coded under one or more themes in the original taxonomy of ethical and professional issues, resulting in a total of 173 coded responses. Professional Duties was the most frequently coded theme (29.2%), followed by Communication (26.4%), Quality of Care (18.8%), Student-specific Issues of Moral Distress (16.7%), Decisions Regarding Treatment (16.0%), and Justice (13.2%). In the virtues taxonomy, 180 total responses were coded from the 144 original responses, and the most frequent virtue coded was Wisdom (23.6%), followed by Respectfulness (20.1%) and Compassion or Empathy (13.9%).Conclusions: Originally developed from students’ clinical experiences in one institution, the Kaldjian taxonomy appears to serve as a useful analytical framework for categorizing a variety of clinical experiences faced by a national sample of medical students. This study also supports the development of virtue-based programs that focus on cultivating the virtue of wisdom in the practice of medicine.
Synthesis of nearly spherical AlN particles by an in-situ nitriding combustion route
Authors: Wei Zhilei , Li Kang , Ge Bangzhi , Guo Chaowei , Xia Hongyan , Guo Yajie , Shi Zhongqi 
Published: October 2020

Abstract
Spherical AlN powders with micrometer size have attracted great attention owing to their good fluidity and dispersity. However, the industrial preparation methods usually require high temperature and long soaking time, which lead to the high cost and limit the wide application of the products. Herein, nearly spherical AlN particles with the average size of 2.5 μm were successfully synthesized via an in-situ combustion synthesis method. The effect of N 2 pressure, NH 4 Cl content and Al particle size on the combustion reaction procedure, phase composition and microstructure of the products was systematically investigated. The results showed that the decreased N 2 pressure, increased NH 4 Cl content and Al particle size led to the decreasing of combustion temperature and speed, which further affected the morphology of the products. As a result, low N 2 pressure (0.2 MPa), a small amount of NH 4 Cl (0.5 wt%) and fine Al particles (~2.5 μm) contributed to a moderate combustion temperature and facilitated the formation of nearly spherical AlN particles. In addition, based on the gas-releasing assisted quenching experiments and thermo-kinetic analysis, a two-step growth mechanism for the nearly spherical AlN particles was rationally proposed. The present method shows the advantages of low cost and high efficiency for preparing nearly spherical AlN particles, which can be used as raw materials for electronic substrates and fillers for packaging materials.
Exosomes from adipose tissue-derived mesenchymal stem cells ameliorate histone-induced acute lung injury by activating the PI3K/Akt pathway in endothelial cells.
Authors: Mizuta Yukie , Akahoshi Tomohiko , Guo Jie , Zhang Shuo , Narahara Sayoko , Kawano Takahito , Murata Masaharu , Tokuda Kentaro , Eto Masatoshi , Hashizume Makoto , Yamaura Ken 
Published: October 2020

Abstract
BackgroundMesenchymal stem cells (MSCs), including adipose-derived mesenchymal stem cells (ADSCs), have been shown to attenuate organ damage in acute respiratory distress syndrome (ARDS) and sepsis; however, the underlying mechanisms are not fully understood. In this study, we aimed to explore the potential roles and molecular mechanisms of action of ADSCs in histone-induced endothelial damage. MethodsMale C57BL/6N mice were intravenously injected with ADSCs, followed by histones or a vehicle. The mice in each group were assessed for survival, pulmonary vascular permeability, and histological changes. A co-culture model with primary human umbilical vein endothelial cells (HUVECs) exposed to histones was used to clarify the paracrine effect of ADSCs. Overexpression and inhibition of miR-126 ADSCs were also examined as causative factors for endothelial protection.ResultsThe administration of ADSCs markedly improved survival, inhibited histone-mediated lung hemorrhage and edema, and attenuated vascular hyper-permeability in mice. ADSCs were engrafted in the injured lung and attenuated histone-induced endothelial cell apoptosis. ADSCs showed endothelial protection (via a paracrine effect) and Akt phosphorylation in the histone-exposed HUVECs. Notably, increased Akt phosphorylation by ADSCs was mostly mediated by exosomes in histone-induced cytotoxicity and lung damage. Moreover, the expression of miR-126 was increased in exosomes from histone-exposed ADSCs. Remarkably, the inhibition of miR-126 in ADSCs failed to increase Akt phosphorylation in histone-exposed HUVECs. ConclusionADSC-derived exosomes may exert protective effects on endothelial cells via activation of the PI3K/Akt pathway.
Efficient isolation of protoplasts from rice calli with pause points and its application in transient gene expression and genome editing assays
Authors: Poddar Snigdha , Tanaka Jaclyn , Cate Jamie H. D. , Staskawicz Brian , Cho Myeong-Je 
Published: October 2020

Abstract
Background: An efficient in vivo transient transfection system using protoplasts is an important tool to study gene expression, metabolic pathways, and multiple mutagenesis parameters in plants. Although rice protoplasts can be isolated from germinated seedlings or cell suspension culture, preparation of those donor tissues can be inefficient, time-consuming, and laborious. Additionally, the lengthy process of protoplast isolation and transfection needs to be completed in a single day. Results: Here we report a protocol for the isolation of protoplasts directly from rice calli, without using seedlings or suspension culture. The method is developed to employ discretionary pause points during protoplast isolation and before transfection. Protoplasts maintained within a sucrose cushion partway through isolation, for completion on a subsequent day, per the first pause point, are referred to as S protoplasts. Fully isolated protoplasts maintained in MMG solution for transfection on a subsequent day, per the second pause point, are referred to as M protoplasts. Both S and M protoplasts, 1 day after initiation of protoplast isolation, had minimal loss of viability and transfection efficiency compared to protoplasts 0 days after isolation. S protoplast viability decreases at a lower rate over time than that of M protoplasts and can be used with added flexibility for transient transfection assays and time-course experiments. The protoplasts produced by this method are competent for transfection of both plasmids and ribonucleoproteins (RNPs). Cas9 RNPs were used to demonstrate the utility of these protoplasts to assay genome editing in vivo. Conclusion: The current study describes a highly effective and accessible method to isolate protoplasts from callus tissue induced from rice seeds. This method utilizes donor materials that are resource-efficient and easy to propagate, permits convenience via pause points, and allows for flexible transfection days after protoplast isolation. It provides an advantageous and useful platform for a variety of in vivo transient transfection studies in rice.
Intrinsic heterogeneity of mossy cells mediates the differential crosstalk between the dorsal and ventral hippocampus
Authors: Jeong Minseok , Jang Jin-Hyeok , Oh Seo-Jin , Choe Young-Shik , Park Jeongrak , Lee Junseop , Park Junwoo , Lee Yaebin , Hwang Sehyeon , Kim Jong , Lee Jusung , Ha Chang Man , Choi Yura , Kim Sang Ryong , Nishi Akinori , Greengard Paul , Oh Yong-Seok 
Published: October 2020

Abstract
Glutamatergic mossy cells (MCs) are responsible for the associational and commissural connectivity of the dentate gyrus. MCs are widely distributed along the dorsoventral axis, but potential heterogeneity within MCs is scarcely explored. Here, we showed that MCs consist of two subpopulations which differ in their neuronal properties and functions. We discovered that MCs, depending on their dorsoventral location, extend distinct axonal projections in the molecular layers. Comparative transcriptional profiling of dorsal and ventral MCs revealed different neurobiological characteristics in axon guidance and synapse assembly. Despite common activation by external stimuli, dorsal MCs, but not ventral MCs, provide net inhibitory control on granule cells across the longitudinal axis. Furthermore, dorsal MC inhibition, unlikely that of ventral MCs, increases behavioral anxiety and disables rapid contextual discrimination. Collectively, dorsoventral heterogeneity of MCs may provide a novel mechanism for functional differentiation as well as distinct association along the longitudinal extent of the hippocampus.
Effect of PCSK9 Inhibitor on Lipoprotein Particles in Patients With Acute Coronary Syndromes
Authors: Li Tingting , Zhang Yingyi , Cong Hongliang 
Published: October 2020

Abstract
Backgrounds: To assess the effects of proprotein convertase subtilisin/kexin type 9 inhibitor (evolocumab) on lipoprotein particles subfractions with Nuclear Magnetic Resonance spectroscopy in patients with acute coronary syndromes.Methods: A total of 99 consecutive patients with ACS and poor lipid control were enrolled and assigned to either the experimental group (n = 54) or the control group (n = 45). The combination therapy of PCSK9 inhibitor (Repatha®, 140mg, q2w) and moderate statin (rosuvastatin, 10 mg, qn) was administered in the experimental group, with statin monotherapy (rosuvastatin, 10 mg, qn) in the control group. The therapeutic effects on lipoprotein particle subfractions were assessed with NMR spectroscopy after eight weeks treatment, and the achievement of LDL-C therapeutic target in both groups were analyzed.Results: In the experimental group, after eight weeks of evolocumab combination treatment, the concentrations of blood lipids (TC, LDL-C and its subfractions [LDL-1 to 6], VLDL-C and its subfractions [VLDL-1 to 5], IDL-C, and HDL-C), lipoprotein particles, and their subfractions (VLDL-P, IDL-P, LDL-P, and its subfractions [LDL-P1 to 6], apoB, and LP(a)) demonstrated therapeutic benefits with statistical significance (P < 0.05). The decrease in total LDL-P concentrations was mainly due to a decreased concentration of small-sized LDL particles (LDL 5+6), which was significantly more prominent than the decrease in medium-sized LDL-P (LDL-P3+4) and large-sized LDL-P (LDL-P1+2) (P < 0.001). According to lipid control target recommended by the latest China Cholesterol Education Program Expert Consensus in 2019, after eight weeks treatment, 96.3% patients in the experimental group and 13.3% in the control group had achieved the LDL-C therapeutic target(P < 0.01).Conclusions: Evolocumab combination treatment for 8 weeks could significantly improve the plasma lipid profiles in ACS patients with poor lipid control, and significantly decrease the concentration of lipoprotein particles which could result in atherosclerosis.