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Oomycetes are diploid eukaryotic microorganisms that seriously threaten sustainable crop production. MicroRNAs (miRNAs) and corresponding natural antisense transcripts (NATs) are important regulators of multiple biological processes. However, little is known about their roles in plant immunity against oomycete pathogens. In this study, we report the identification and functional characterization of miR398b and its cis-NAT, the core-2/I-branching beta-1,6-N-acetylglucosaminyltransferase gene (AtC2GnT), in plant immunity. Gain- and loss-of-function assays revealed that miR398b mediates Arabidopsis thaliana susceptibility to Phytophthora parasitica by targeting Cu/Zn-Superoxidase Dismutase1 (CSD1) and CSD2, leading to suppressed expression of CSD1 and CSD2 and decreased plant disease resistance. We further showed that AtC2GnT transcripts could inhibit the miR398b-CSDs module via inhibition of pri-miR398b expression, leading to elevated plant resistance to P. parasitica. Furthermore, quantitative reverse transcription PCR, RNA ligase-mediated 5'-amplification of cDNA ends (RLM-5' RACE), and transient expression assays indicated that miR398b suppresses the expression of AtC2GnT. We generated AtC2GnT-silenced A. thaliana plants by CRISPR/Cas9 or RNA interference methods, and the Nicotiana benthamiana NbC2GnT-silenced plants by virus-induced gene silencing. Pathogenicity assays showed that the C2GnT-silenced plants were more susceptible, while AtC2GnT-overexpressing plants exhibited elevated resistance to P. parasitica. AtC2GnT encodes a Golgi-localized protein, and transient expression of AtC2GnT enhanced N. benthamiana resistance to Phytophthora pathogens. Taken together, our results revealed a positive role of AtC2GnT and a negative regulatory loop formed by miR398b and AtC2GnT in regulating plant resistance to P. parasitica.

Pyroptosis, as a novel identified programmed cell death, is closely correlated with tumor immunity and shows potential roles in cancer treatment. Discerning a pyroptosis-related gene signature and its correlations with tumor immune microenvironment is critical in head and neck squamous cell carcinoma (HNSCC). Transcriptome data and corresponding clinical data were downloaded from TCGA and GEO databases. Tumor mutation burden (TMB) data were obtained from TCGA database. Firstly, univariate and least absolute shrinkage and selection operator (LASSO) regression analyses were used to construct a six pyroptosis-related gene signature. Kaplan–Meier analysis, receiver operating characteristic (ROC) curves, and principal component analysis (PCA) results verified that the risk model has good performance in predicting the survival. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses revealed that the pyroptosis-related gene signature was immune related. Finally, the immune landscape and immunotherapy sensitivity prediction capabilities of the risk model were further explored. There were close correlations between the overall survival (OS) and various immune cells and immune functions. Single-sample gene set enrichment analysis (ssGSEA) showed that high risk group had decreased expression of various immune cells and lower activities of immune functions. Meanwhile, tumor mutation burden (TMB) data combining risk score could well predict the OS of HNSCC patients. However, tumor immune dysfunction and exclusion (TIDE) analysis revealed that there was no significant difference in the sensitivity to immunotherapies between high and low risk groups. Finally, a nomogram based on risk score and clinicopathological parameters was constructed. And, the risk model demonstrated better sensitivity and specificity than TIDE scores and T-cell-inflamed signature (TIS). In conclusion, although the risk model could not well predict the immune escape and response to immunotherapies, the signature established by pyroptosis-related genes, with better sensitivity and specificity than TIDE scores and TIS signature, could be used for predicting prognosis and immune status of HNSCC patients.

SummaryEpstein-Barr virus (EBV) T/NK-cell lymphoproliferative diseases are characterized by clonal expansion of EBV-infected T or NK cells, including chronic active EBV infection of T/NK-cell type (CAEBV+T/NK), EBV-associated hemophagocytic lymphohistiocytosis (EBV+HLH), extranodal NK/T-cell lymphoma of nasal type (ENKTL), and aggressive NK-cell leukemia (ANKL). However, the role of inherited genetic variants to EBV+T/NK-LPDs susceptibility is still unknown. A total of 171 nonimmunosuppressed patients with EBV+T/NK-LPDs and 104 healthy donors were retrospectively collected and a targeted sequencing study covering 15 genes associated with lymphocyte cytotoxicity was performed. The 94 gene variants, mostly located in UNC13D, LYST, ITK, and PRF1 genes were detected, and mutations covered 28/50 (56.00%) of CAEBV-T/NK, 31/51 (60.78%) of EBV+HLH, 13/28 (46.42%) of ENKTL, and 13/48 (27.09%) of ANKL. Most mutations represented monoallelic and missense. Three-year overall survival rate of patients with CAEBV-T/NK and EBV+HLH was significantly lower in patients with germline mutations than in those without germline mutations (P=0.0284, P=0.0137). Our study provided novel insights into understanding a spectrum of nonimmunosuppressed EBV+T/NK-LPDs with respect to genetic defects associated with lymphocyte cytotoxicity and reminded us that the gene sequencing may be an auxiliary test for diagnosis and risk stratification of EBV+T/NK-LPDs.

Epstein-Barr virus (EBV), characterized as an omnipresent virus, has been found able to infect NK cells and leads to NK-cell type EBV-positive lymphoproliferative diseases (EBV-NK-LPDs). We retrospective analyzed 202 EBV-NK-LPDs (including 64 CAEBV-NK, 27 aggressive natural killer-cell leukemia (ANKL), and 111 extranodal NK/T-cell lymphoma (ENKTL)) patients' relationships between EBV DNA copies laboratory test results and clinical features. In CAEBV-NK cohort, EBV DNA loads in either plasma or PBMCs had significant differences between the active state and the inactive state. Receiver operating characteristic curves were used to measure the diagnosis accuracy of EBV DNA copies. After comparing the area under the curve, EBV DNA loads in plasma had significantly higher accuracy in distinguishing disease activation than in PBMCs. Therefore, we propose redefining CAEBV-NK diagnosis criteria as increased EBV DNA copies in plasma (over 7.1 × 10

BackgroundCalcium-dependent protein copine 8 (CPNE8), a new member of Copine family, emerges important roles in malignancies. However, the expression and potential role of CPNE8 in pan cancer remain unclear. Methods: Pan cancer analysis of CPNE8 expression in 33 types of cancers were based on data downloaded from the University of California Santa Cruz (UCSC) Xena database. The clinical significances of CPNE8 were analyzed using Cox regression analysis and Kaplan Meier survival analysis. Correlations between CPNE8 expression and tumor immune microenvironment were assessed using ESTIMATE algorithm, CIBERSORT analysis and online database TISIDB. The correlations between CPNE8 expression and tumor mutation burden (TMB), microsatellite instability (MSI) status and response to immune checkpoint inhibitors (ICIs) were further explored. Moreover, competitive endogenous RNA (ceRNA) networks related to CPNE8 and its correlations with immune cell related genes were further studied in stomach adenocarcinoma (STAD).Results: Differential expression of CPNE8 were identified in 14 types of cancers. Differential expression of CPNE8 could predict tumor stage or prognosis of various cancer patients. Pan cancer analysis indicated that CPNE8 demonstrated oncogenic role in STAD, head and Neck squamous cell carcinoma (HNSC) and esophageal carcinoma (ESCA), and tumor suppressive role in thyroid carcinoma (THCA). These four cohorts were selected for further study due to their consistent results in expression and prognostic values. Gene set enrichment analysis (GSEA) revealed that CPNE8 might regulate several key immune-related signaling pathways. Furthermore, CPNE8 expression was significantly correlated with the infiltration of different subtypes of immune cells, TMB and MSI. Moreover, PWAR5 and LIFR-AS1 could sponge miR-222-3p as ceRNAs to regulate the expression of CPNE8 in STAD. In addition, CPNE8 was significantly correlated with the expression of immune cell related genes, such as CD8 T cells, macrophages, B cells and dendritic cells in STAD.Conclusion: This study demonstrated that CPNE8 could serve as a promising biomarker associated with the prognosis and immune microenvironment in several malignancies.

Automatic seizure detection technology has important implications for reducing the workload of neurologists for epilepsy diagnosis and treatment. Due to the unpredictable nature of seizures, the imbalanced classification of seizure and nonseizure data continues to be challenging. In this work, we first propose a novel algorithm named the borderline nearest neighbor synthetic minority oversampling technique (BNNSMOTE) to address the imbalanced classification problem and improve seizure detection performance. The algorithm uses the nearest neighbor notion to generate nonseizure samples near the boundary, then determines the seizure samples that are difficult to learn at the boundary, and lastly selects seizure samples at random to be used in the synthesis of new samples. In view of the characteristic that electroencephalogram (EEG) signals are one-dimensional signals, we then develop a 1D-MobileNet model to validate the algorithm’s performance. Results demonstrate that the proposed algorithm outperforms previous seizure detection methods on the CHB-MIT dataset, achieving an average accuracy of 99.40%, a recall value of 87.46%, a precision of 97.17%, and an F1-score of 91.90%, respectively. We also had considerable success when we used additional datasets for verification at the same time. Our algorithm’s data augmentation effects are more pronounced and perform better at seizure detection than the existing imbalanced techniques. Besides, the model’s parameters and calculation volume have been significantly reduced, making it more suitable for mobile terminals and embedded devices.

7532 Background: Long-term outcome is unfavorable for patients with relapsed/refractory (r/r) classical Hodgkin lymphoma (cHL) who are resistant to salvage chemotherapy, even followed by autologous stem cell transplantation (ASCT). Long-term survival is most likely achieved in patients who are sensitive to salvage therapy and have a complete response (CR) before ASCT, while outcome is poor in patients with residual disease. Although CD30-directed chimeric antigen receptor (CAR30) T-cell therapy induces high response rates in these patients, the duration of response is limited. Methods: We conducted an open-label, single-center and single-arm pilot study to explore the safety and efficacy of ASCT in tandem with CAR30 T-cell infusion in r/r CD30+ lymphoma (Chinese Clinical Trial Registry, number ChiCTR 2100053662). Patients will receive BEAM as preconditioning. HSCs are infused at day zero and followed by the infusion of 3rd generation CAR30 T cells in the next week. Between June 1, 2019 to May 1, 2021, 6 patients enrolled, including 5 with cHL and 1 with anaplastic lymphoma kinase-negative anaplastic large cell lymphoma (ALCL). The median age was 24 years. No patient had a prior history of ASCT. Three patients (50.0%) relapsed ≥ 2 times and 3 patients (50.0%) had primary refractory diseases. All had a Deauville score of 4 or 5, and 5 patients (83.3%) had a stable or progressive disease (SD/PD) to previous treatments at enrollment. Results: CD34+ cells were infused at day zero with a median dosage of 3.9 (IQR, 3.2-6.1) × 106 /kg and followed by the infusion of CAR30 T cells with a median dosage of 7.6 (IQR, 5.5-9.7 ) ×106 /kg at +2 to +6 days. Cytopenias represented the most commonly severe advers events (≥ grade 3). The engraftments of neutrophil or platelet occured at a median time of 13.5 (IQR,12.3-14.0 ) days or 11.5 (IQR, 11.0-12.8) days, respectively, suggesting rapid multilineage engraftments post-ASCT. Cytokine release syndrome (CRS) was observed in 5 (83.3%) patients, all of which were grade 1. No neurotoxicity or severe infection was observed. At month 3 after HSC infusion, all patients achieved objective responses, including 5 (83.3%) with a CR and 1 with a patial response (PR). With a median follow-up of 18.2 (range, 9.9-32.1) months, the median PFS and OS were not reached. At January 31, 2022, the data-cutoff date, all patients maintained their responses and remained alive without disease relapse or progression. Of note, responses were sustained in all 5 patients who had a SD/PD at enrollment. Conclusions: Our work demonstrates tandem administration of ASCT and CAR30 T-cell therapy was well-tolerated and highly active in r/r HL and ALCL, even in PET-positive or chemorefractory patients who were expected to have inferior outcome after ASCT, although further large-scale validation in prospective clinical trial is warranted. Clinical trial information: ChiCTR2100053662.

Highlights • NSE expression was positively correlated with distant metastasis of SCLC. • NSE promotes migration, invasion and EMT process of SCLC cells. • Wnt/β-catenin signaling pathway is activated in the NSE-overexpressing SCLC cells. • NSE interacts with β-catenin and inhibits the degradation of β-catenin., Neuron-specific enolase (NSE) has been used as a specific biomarker for small cell lung cancer (SCLC) patients. Nevertheless, the biological function and mechanism of NSE in SCLC are still unclear. In this study, we clarified the role of NSE in the progression of SCLC and found that NSE expression was positively correlated with distant metastasis. Functional analysis showed that overexpression of NSE promoted migration and invasion of SCLC cells. Mechanism analysis showed that NSE overexpression induced epithelial-mesenchymal transition (EMT) of SCLC cells. Moreover, overexpression of NSE increased the protein expression of β-catenin and its downstream target genes, and silencing β-catenin eliminated NSE-mediated cell migration, invasion and EMT process. Furthermore, NSE interacted with β-catenin and inhibited the degradation of β-catenin. Besides, the animal experiments also indicated that NSE could promote the EMT process and distant metastasis of SCLC cells in vivo. In summary, our results revealed that NSE could promote the EMT process of SCLC cells by activating the Wnt/β-catenin signaling pathway, thereby promoting cell migration, invasion and distant metastasis, which might serve as a potential target for the therapy of SCLC patients.

In vitro construction is a major trend involved in cartilage regeneration and repair. Satisfactory in vitro cartilage regeneration depends on a suitable culture system. Current chondrogenic culture systems with a high content of transforming growth factor beta-1 effectively promote cartilaginous extracellular matrix (ECM) production but inhibit chondrocyte survival. As is known, inhibition of the c-Jun N-terminal kinase (JNK) signaling pathway acts in blocking the progression of osteoarthritis by reducing chondrocyte apoptosis and cartilage destruction. However, whether inhibiting JNK signaling resists the inhibitory effect of current chondrogenic medium (CM) on cell survival and affects in vitro auricular cartilage regeneration (including cell proliferation, ECM synthesis, and degradation) has not been investigated. In order to address these issues and optimize the chondrogenic culture system, we generated a three-dimensional in vitro auricular cartilage regeneration model to investigate the effects of SP600125 (a JNK-specific inhibitor) on chondrocyte proliferation and ECM metabolism. SP600125 supplementation efficiently promoted cell proliferation at both cellular and tissue levels and canceled the negative effect of our chondrogenic culture system on cell survival. Moreover, it significantly inhibited ECM degradation by reducing the expressions of tumor necrosis factor-alpha, interleukin-1-beta, and matrix metalloproteinase 13. In addition, SP600125 inhibited ECM synthesis at both cellular and tissue levels, but this could be canceled and even reversed by adding chondrogenic factors; yet this enabled a sufficient number of chondrocytes to be retained at the same time. Thus, SP600125 had a positive effect on in vitro auricular cartilage regeneration in terms of cell proliferation and ECM degradation but a negative effect on ECM synthesis, which could be reversed by adding CM. Therefore, a combination of SP600125 and CM might help in optimizing current chondrogenic culture systems and achieve satisfactory in vitro cartilage regeneration by promoting cell proliferation, reducing ECM degradation, and enhancing ECM synthesis.

BackgroundNeuron-specific enolase (NSE) has been used as a specific biomarker for small cell lung cancer (SCLC) patients. Nevertheless, the biological function and mechanism of NSE in SCLC are still unclear. MethodsWe conducted an investigation in SCLC patients and then analyzed the association between NSE concentration and the distant metastasis. The migration and invasion were analyzed using Wound-healing and Transwell assays. The proteins and mRNA levels were measured by western blot and qRT-PCR. Furthermore, we conducted Gene Set Enrichment Analysis (GSEA) to explore potential signaling pathways. The interaction between NSE and β-catenin was verified through CO-IP experiments. In addition, animal experiments were performed in tumor metastasis models of nude mice.ResultIn this study, we clarified the role of NSE in the progression of SCLC and found that NSE expression was positively correlated with distant metastasis. Functional analysis showed that overexpression of NSE promoted migration, invasion and metastasis of SCLC cells, while knocking down NSE inhibited these effects. Mechanism analysis showed that NSE overexpression induced epithelial-mesenchymal transition (EMT) of SCLC cells, while knocking down NSE inhibited this effect. Moreover, overexpression of NSE increased the protein expression of β-catenin and its downstream target genes, and silencing β-catenin eliminated NSE-mediated cell migration, invasion and EMT process. Furthermore, NSE interacted with β-catenin and inhibited the degradation of β-catenin. Besides, the animal experiments also indicated that NSE could promote the EMT process and tumor distant metastasis of SCLC cells by activating Wnt/β-catenin pathway in vivo.ConclusionOur results revealed that NSE could promote EMT process of SCLC cells by activating the Wnt/β-catenin signaling pathway, thereby promoting migration, invasion and distant metastasis, which might serve as a potential target for the therapy of SCLC patients.

Background This study aimed to investigate the preventive effect of translocator protein 18kDa (TSPO) ligand PK11195 on amnestic mild cognitive impairment (aMCI), as well as its influence on astrocytes, in order to identify effective ways to prevent aMCI. Methods Male SD rats were randomly divided into control group (n=10), aMCI group (n=10), PK11195 group (n=10), PK11195 + D-gal group (n=10). The preventive effect of PK11195 on aMCI in rats was evaluated. The cognitive function of rats in four different treatment groups was determined using the Morris water maze (MWM), as well as whole-brain pathology and immunofluorescence of rat brain tissue. Results The results of the MWM behavioral test showed that rats pre-treated with PK11195 had improved escape latency and a higher number of platform crossings compared with the aMCI model rats. PK11195 was also shown to prevent the D-galactose (D-gal)-induced senescence of pyramidal cells in the hippocampal CA1 region and to inhibit the apoptosis of astrocytes. At the same time, compared with the aMCI model rats, the TSPO in the brain tissue of rats pretreated with PK11195 had a lower distribution density. Conclusions Our results prove that PK11195 can effectively prevent D-gal-induced decline of learning and memory function as well as inhibit abnormal changes of related cells.

Background. Long noncoding RNA-based prognostic biomarkers have demonstrated great potential in the diagnosis and prognosis of cancer patients. However, systematic assessment of a multiple lncRNA-composed prognostic risk model is lacking in stomach adenocarcinoma (STAD). This study is aimed at constructing a lncRNA-based prognostic risk model for STAD patients. Methods. RNA sequencing data and clinical information of STAD patients were retrieved from The Cancer Genome Atlas (TCGA) database. Differentially expressed lncRNAs (DElncRNAs) were identified using the R software. Univariate and multivariate Cox regression analyses were performed to construct a prognostic risk model. The survival analysis, C-index, and receiver operating characteristic (ROC) curve were employed to assess the sensitivity and specificity of the model. The results were verified using the GEPIA online tool and our clinical samples. Pearson correlation coefficient analysis, Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment were performed to indicate the potential biological functions of the selected lncRNA. Results. A total of 1917 DElncRNAs were identified from 343 cases of STAD tissues and 30 cases of noncancerous tissues. According to univariate and multivariable Cox regression analyses, four DElncRNAs (AC129507.1, LINC02407, AL022316.1, and AP000695.2) were selected to establish a prognostic risk model. There was a significant difference in the overall survival between high-risk patients and low-risk patients based on this risk model. The C-index of the model was 0.652. The area under the curve (AUC) for the ROC curve was 0.769. GEPIA results confirmed the expression and prognostic significance of AP000695.2 in STAD. Our clinical data confirmed that upregulated expression of AP000695.2 was correlated with the T stage, distant metastasis, and TNM stage in STAD. GO and KEGG analyses demonstrated that AP000695.2 was closely related to the tumorigenesis process. Conclusions. In this study, we constructed a lncRNA-based prognostic risk model for STAD patients. Our study will provide novel insight into the diagnosis and prognosis of STAD patients.

Noble metal materials are widely employed as benchmark electrocatalysts to achieve electrochemical water splitting which comprises of hydrogen evolution reaction (HER) and oxygen evolution reaction (OER). However, the high cost and scarcity limit the wide ranging commercial applications of noble metal-based catalysts. Development of noble metal-free two dimensional (2D) carbon-based materials can not only reduce the consumption of noble metals, but also create materials with the characteristics of high active surface area, abundance, easy functionalization, and chemical stability, which may carve a way to promising electrochemical water splitting. In this review, noble metal-free 2D carbon-based electrocatalysts, including heteroatom (B, S, N, P, F, and O) doped graphene, 2D porous carbons modified with heteroatoms and/or transition metals, and 2D carbon-based hybrids are introduced as cost-effective alternatives to the noble metal-based electrocatalysts with comparable efficiencies to conduct HER, OER, and overall water splitting. This review emphasizes on current development in synthetic strategies and structure–property relationships of noble metal-free 2D carbon-based electrocatalysts, together with major challenges and perspectives of noble metal-free 2D carbon-based electrocatalysts for further electrochemical applications.

// Bin Xu 1 , Yi Tang 1 , Jianfeng Zhou 1 , Peiling Zhang 1 and Huijun Li 2 1 Department of Hematology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China 2 Department of Clinical Laboratory, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China Correspondence to: Yi Tang, email: tangyi_100@qq.com Keywords: abnormal serum free light chain ratio, cut-off value, malignant plasma diseases, MGUS, reactive plasma diseases Received: April 04, 2017 Accepted: June 11, 2017 Published: July 19, 2017 ABSTRACT Objective: To analyze the spectrum of abnormal serum free light chain ratio (sFLC κ/λ ratio), and to redefine the range of sFLC κ/λ ratio, so as to achieve hierarchical diagnosis of diseases with abnormal sFLC κ/λ ratio, resulting in the increased sensitivity and specificity in the diagnosis of monoclonal plasma diseases. Methods: Enrolled 1,340 patients with abnormal sFLC κ/λ ratio ( 1.65) were grouped: (1) group A: malignant plasma diseases; (2) group B: monoclonal gammopathies of undetermined significance (MGUS); (3) group C: reactive plasma diseases. These patients were further divided by renal function eGFR 60 ml/min/1.73m 2 to eliminate renal diseases influencing the results. Statistical analyses was performed by using SPSS 22 software. Results: When sFLC κ/λ ratio >3.49 and eGFR >60ml/min/1.73m 2 , the sensitivity and specificity of the diagnosis of malignant plasma diseases were 86.1% and 94.0%, respectively. When sFLC κ/λ ratio >2.89 and eGFR

Three novel complexes, namely {[Zn3(L)2(SO4)2(H2O)4]·2H2O}n (1), {[Cd(L)(OAc)]}n (2) and [Hg2(L)(I)3]2 (3) (HL = N′ - isonicotinoylpyrimidine -2- carbohydrazonamide), have been synthesized and characterized by elemental analyses, infrared spectra and single-crystal X-ray diffraction analyze. The 1D complex 1 and complex 2, and the binuclear complex 3 are expanded to 3D networkers by the strong hydrogen bonds and π-π stacking interactions. The photocatalytic degradation of methylene blue (MB) results indicate that complexes 1–3 are excellent candidates as photocatalysts in decomposing MB with the presence of H2O2. In addition, the luminescent properties of these three complexes have been studied in the solid state.

Background High loads of Epstein-Barr virus (EBV) in peripheral blood mononuclear cells (PBMCs) can be indicative of a broad spectrum of diseases, ranging from asymptomatic infection to fatal cancers. Methods We retrospectively investigated the EBV-infected cell types in PBMCs among 291 patients. Based on EBV-infected cell types, the clinical features and prognoses of 93 patients with EBV-associated (EBV+) T/natural killer (NK)–cell lymphoproliferative diseases (LPDs) T/NK-LPDs) were investigated over a 5-year period. Results Although B-cell-type infection was found in immunocompromised patients and patients with asymptomatic high EBV carriage, infectious mononucleosis, EBV+ B-cell LPDs and B-cell lymphomas, T-cell, NK-cell or multiple-cell-type infection in immunocompetent hosts were highly suggestive of EBV+ T/NK-LPDs, EBV+ T/NK-cell lymphomas, and aggressive NK-cell leukemia. Patients with non–B-cell infection had a poorer prognosis than those with B-cell-type infection. In our cohort, 79.6% of patients with EBV+ T/NK-LPDs were >18 years old, and NK cells were identified as EBV-infected cell type in 54.8%. Nearly half of patients with EBV+ T/NK-LPDs had genetic defects associated with immunodeficiency. However, hemophagocytic lymphohistiocytosis, and not genetic defects, was the only parameter correlated with poor prognosis of EBV+ T/NK-LPDs. Conclusions Determination of EBV-infected cell types among PBMCs is a valuable tool for the differential diagnosis of EBV+ hematological diseases. In this study, determination of Epstein-Barr virus-infected cell types in peripheral blood mononuclear cells of 291 patients with high Epstein-Barr virus loads were retrospectively investigated, which indicate it is a valuable tool for Epstein-Barr virus-associated hematological diseases.

// Yang Cao 1, * , Tao Zhu 1, * , Peiling Zhang 1 , Min Xiao 1 , Shuhua Yi 2 , Yan Yang 1 , Qinlu Li 1 , Shaoping Ling 3 , Yafei Wang 4 , Lili Gao 1 , Li Zhu 1 , Jue Wang 1 , Na Wang 1 , Liang Huang 1 , Peihong Zhang 2 , Qiongli Zhai 4 , Lugui Qiu 2 , Jianfeng Zhou 1, 2 1 Department of Hematology & Cancer Biology Research Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, P.R. China 2 State Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin, P.R. China 3 Laboratory of Genome Variations and Precision Bio-Medicine, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, P.R. China 4 Tianjin Cancer Hospital, Tianjin Medical University, Tianjin, P.R. China * These authors have contributed equally to this work Correspondence to: Jianfeng Zhou, email: jfzhou@tjh.tjmu.edu.cn Lugui Qiu, email: drqiu99@medmail.com.cn Keywords: TP53, CD58, mutation, copy number loss, DLBCL Received: September 24, 2015 Accepted: September 02, 2016 Published: November 4, 2016 ABSTRACT The advent of next generation sequencing (NGS) technologies has expedited the discovery of novel genetic lesions in DLBCL. The prognostic significance of these identified gene mutations is largely unknown. In this study, we performed NGS for the 27 genes most frequently implicated in 196 patients. Interestingly, TP53 mutations were found to be significantly more common in DLBCL with MYC translocations ( r = 0.446, P = 0.034). While no gene mutation was found to be more prevalent in patients with DLBCL with bone marrow involvement, MYD88 mutations were more common in primary DLBCL of the CNS or testis. To evaluate the prognostic significance of the abnormalities of these 27 genes, a total of 165 patients with newly diagnosed DLBCL, NOS were included in a multivariate survival analysis. Surprisingly, in addition to the TP53 mutation, CD58 mutation was found to predict poor clinical outcome. Furthermore, copy number loss of CD58 or TP53 was also identified to be an independent negative prognostic factor. Our results have uncovered the previously unknown critical impact of gene mutations on the prognosis of DLBCL and are fundamentally important for the future design of tailored therapy for improved clinical outcomes.

Background In the context of booming e-commerce, logistics service is an opportunity as well as a challenge for fresh products e-retailers and their logistics service providers (LSPs). In fact, they offered various fancy logistics services, like evening home delivery, same-day delivery and recyclable packaging. However, it will lead to a high cost. So, it is necessary for them to understand consumer preferences for logistics services to obtain their satisfaction and loyalty as well as to decrease the logistics costs. Purpose The purpose of this study is to investigate consumer preferences for logistics services within online retailing of fresh products in Sweden. Method To fulfill the research purpose, a quantitative research is conducted by employing the full-profile conjoint analysis. The quantitative data in a raw form were collected from the questionnaire, which has investigated 161 respondents in Sweden for rating nine logistics service profiles. To reduce the respondents’ evaluation task, the profiles are generated by orthogonal design in SPSS 24.0. The data analysis is conducted by conjoint analysis module in the software. Conclusion The results show that free home delivery is the preferred choice and green packaging material is much more attractive than ordinary one. Delivery speed does not appear to be so important comparing with the other attributes, which means the respondents could accept the delivery within one week. To extent, the respondents are very cost conscious, but they are still willing to make trade off for the premium logistics services. For example, they could accept 1-50 kronor delivery fee when they select home delivery with green package marticals. Therefore, e-retailers and their LSPs can focus on how to balance free shipping and certain delivery costs in different contexts.

A system identification of the speech signal model is presented and discussed which bases on the fuzzy system model. In order to overcome the problem that the approximation property of the past fuzzy clustering algorithms is not ideal when identifying systems, a new Fuzzy Clustering Neural Network (FCNN) is put forward. In this model, presenting Continuous Hidden Markov Model (CHMM) each state as a fuzzy system and using continuous frames as the system input, FCNN calculates each state’s output probability with improving fuzzy clustering identification algorithm. It not only can efficiently import the frames correlation information of the speech signal, but also can overcome the assumption that traditional CHMM each state’s output is mixture Gaussian probability density function. Speech signal system identification which bases on the new model is realized. The experimental results have validated the efficiency of the FCNN that proposed in this paper.

// Xing Xin 1, * , Peiling Zhang 1, * , Xing Fu 1 , Xia Mao 1 , Fankai Meng 1 , Ming Tian 1 , Xiaojian Zhu 1 , Hanying Sun 1 , Li Meng 1 and Jianfeng Zhou 1 1 Department of Hematology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, P. R. China * These authors have contributed equally to this work Correspondence to: Xiaojian Zhu, email: zhuxiaojian@hust.edu.cn Jianfeng Zhou, email: jfzhou@tjh.tjmu.edu.cn Keywords: phosphate-buffered saline, saline, flow cytometry, microvesicles Received: September 20, 2016 Accepted: February 20, 2017 Published: March 07, 2017 ABSTRACT Microvesicles (MVs) are carriers of molecular and oncogenic signatures present in subsets of tumor cells and tumor-associated stroma, and a focus of cancer research. Although methods to detect MVs are mature, we were concerned that the buffer used could lead to false results when quantitating MVs by flow cytometry. In this work,we detected MVs by flow cytometry withthree different solutions: water, saline, and phosphate-buffered saline (PBS). The results demonstrated that PBS, when reacted with annexin V binding buffer, produced nano-sized vesicles even when there were no MVs in the sample. No similar events occurred in the saline and water groups ( P < 0.01). Annexin V positive rate increased significantly when PBS was used as the buffer, compared to saline and water. These false negative results were also observed when we quantified some markers of MVs such as CD3 and CD19. A probable explanation for these findings is the production of insoluble Ca(H 2 PO 4 ) 2 or Ca 3 PO 4 from calcium in the binding buffer and phosphate in PBS. Thus, considering the osmotic pressure of water, we suggest that saline is a more suitable buffer when counting MVs by flow cytometry.

This paper presents bio-oil preparation by direct liquefaction of Dunaliella tertiolecta (D. tertiolecta) with sub/supercritical ethanol-water as the medium in a batch autoclave with high temperature and high pressure. The results indicated that ethanol and water showed synergistic effects on direct liquefaction of D. tertiolecta. The maximum bio-oil yield was 64.68%, with an optimal D. tertiolecta conversion of 98.24% in sub/supercritical ethanol-water. The detailed chemical compositional analysis of the bio-oil was performed using an EA, FT-IR, and GC-MS. The empirical formulas of the bio-oil obtained using the ethanol-water co-solvent (40%, v/v) and sole water as the reaction medium were CH(1.52)O(0.14)N(0.06) and CH(1.43)O(0.23)N(0.09), with calorific values of 34.96 and 29.80 MJ kg(-1), respectively. XPS and SEM results showed that ethanol-water is a very effective reaction medium in the liquefaction. A plausible reaction mechanism of the main chemical component in D. tertiolecta is proposed based on our results and the literatures.

Eosinophilia, such as familial eosinophilia (FE), idiopathic eosinophilia and reactive eosinophilia, is characterized by a sustained overproduction of peripheral blood eosinophils with or without end-organ damage. Germline variants in cytokine and eosinophil function genes are known to contribute to the development of the disease, suggesting common genetic basis may be shared among FE and sporadic eosinophilia. For example, FE is a rare autosomal dominant disorder with susceptible locus mapped to the chromosome 5q region in one American famliy (Rioux et al., 1998). This region (5q31-33) contains several cytokine genes which play functional roles in the development of eosinophils. Germline mutation in JAK1, which is involved in cytokine signaling, has been found in inherited eosinophilia (Kate et al., 2016). Furthermore, genetic variants in genes associated with eosinophilopoietic function (CCR3,CCL5 etc) have been identified in several idiopathic eosinophilia (Lee et al.,2017). Despite of these findings, most cases of eosinophilia have no identifiable mutations. In this new study, we collected a Chinese three-generation pedigree with documented FE. We aimed to investigate novel genetic variants of FE in this pedigree, and found that APP (amyloid-β precursor protein) might be a novel predisposition gene associated with FE. Firstly, we performed Whole-Genome sequencing analysis on isolated eosinophils from peripheral blood of 3 patients and 3 unaffected family members. Point mutations and small indels were selected as potential pathogenic variants if they met the following criteria: 1. heterozygous mutations 2. not previously described 3. predicted to be deleterious or located in regulatory region. As a result of this analysis, 109 mutated genes were considered as candidates, of which APP was the only gene carrying nonsynonymous mutation in the coding region. APP encodes amyloid-β precursor protein that can generate a number of functional peptides. A deleterious missense mutation (c.C470G, p.T157R) was found in the protein binding domain of APP. This mutation was verified by Sanger sequencing in all 3 FE patients. APP has been shown to regulate gene expression (Kögel et al., 2005). As APP knock-out mouse exhibits decreased mRNA expression levels of several cytokines (IL-6, IL-10 and IL-1β) (Carrano et al., 2015), and IL-1β is associated with eosinophil activation (Esnault et al., 2013), we pursued further on APP as a potential eosinophilia predisposition gene. Secondly, we explored transcriptional changes that may be associated with germline mutated APP in eosinophilia patients. We adopted RNA-seq analysis on isolated eosinophils from peripheral blood of 3 patients and 3 controls. A total of 2035 (up:1249, down:786) differentially expressed genes (DEGs) were identified in sorted patient eosinophils as compared to controls (FDR < 0.01). To functionally classify these genes, we conducted pathway analysis and identified "Lysosome", "Chemokine signaling", "Cytokine-cytokine receptor interaction" as significant pathways associated with DEGs. In addition, we found that 51 DEGs were overlapped with previously documented eosinophil-associated genes based on IPA Knowledge Base (N=324). We further determined whether these 51 eosinophil-associated DEGs are interacted with 109 mutated genes by constructing the protein-protein interaction network (PPI). Remarkably, APP was the only gene that was functional linked to 10 eosinophil-associated proteins, including several cytokines (CXCRs, CCRs), to form a sub-network (Score > 0.9). In contrast, randomly picked 51 genes failed to form PPI network. These results indicated that the interactions between APP and eosinophil-associated proteins were potentially of biological significance. Since the analysis of PPI was based on the prior knowledge, we further conducted co-expression analysis on APP and its interaction proteins using our transcriptome data. Interestingly, we found that APP expression was highly correlated with the mRNA level of several eosinophilia-related cytokines including IL-1β, CCR2,CCR3 and CXCR4 (PCC > 0.8). Collectively, our data show that APP is a promising predisposition gene that may contributes to FE as a cytokine modulator. This study furthers our understanding of the genetic basis of familial eosinophilia and other eosinophilia and provides novel therapeutic targets for this heterogenous disease. Disclosures No relevant conflicts of interest to declare.