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In multiple myeloma, abnormal plasma cells accumulate and proliferate in the bone marrow. Recently, we observed that Runx2, a bone-specific transcription factor, is highly expressed in multiple myeloma cells and is a major driver of multiple myeloma progression in bone. The primary goal of the present study was to identify Runx2-targeting miRNAs that can reduce tumor growth. Expression analysis of a panel of miRNAs in multiple myeloma patient specimens, compared with healthy control specimens, revealed that metastatic multiple myeloma cells express low levels of miR-342 and miR-363 but high levels of Runx2. Reconstituting multiple myeloma cells (CAG) with miR-342 and miR-363 reduced the abundance of Runx2 and the expression of metastasis-promoting Runx2 target genes RANKL and DKK1, and suppressed Runx2 downstream signaling pathways Akt/β-catenin/survivin, which are required for multiple myeloma tumor progression. Intravenous injection of multiple myeloma cells (5TGM1), stably overexpressing miR-342 and miR-363 alone or together, into syngeneic C57Bl/KaLwRij mice resulted in a significant suppression of 5TGM1 cell growth, decreased osteoclasts and increased osteoblasts, and increased antitumor immunity in the bone marrow, compared with mice injected with 5TGM1 cells expressing a miR-Scramble control. In summary, these results demonstrate that enhanced expression of miR-342 and miR-363 in multiple myeloma cells inhibits Runx2 expression and multiple myeloma growth, decreases osteolysis, and enhances antitumor immunity. Thus, restoring the function of Runx2-targeting by miR-342 and miR-363 in multiple myeloma cells may afford a therapeutic benefit by preventing multiple myeloma progression.Implications: miR-342 and miR-363–mediated downregulation of Runx2 expression in multiple myeloma cells prevents multiple myeloma progression. Mol Cancer Res; 16(7); 1138–48. ©2018 AACR.

Antibodies used for Western blotting and FACS.

5TGM1 cells stably overexpressing miR-342 and miR-363 were established by lentiviral vector transduction.

Objective To establish a workflow for isolating single trophectoderm (TE) and inner cell mass (ICM) cells and to simultaneously evaluate these cells for copy number variation (CNV) as well as methylome development. Design Experimental. Setting Academic medical center. Patient(s) Donated genetically abnormal blastocysts. Intervention(s) Single cells were isolated, followed by bisulfite conversion and sequencing to identify CNV and methylome profiles. Main Outcome Measure(s) CNV and methylation profiling. Result(s) Two embryos were dissociated, isolating 46 single cells, with 17 ICM and 12 TE cells selected for further downstream analysis. Chromosome ploidies and embryo sex were concordant with the results from conventional aneuploidy testing. In 3 of the 29 cells, additional aneuploidies were discovered, indicating possible mosaicism undetected by routine preimplantation genetic testing for aneuploidy. CpG methylation frequency was higher in ICM cells compared with TE cells (44.3% vs. 32.4%), respectively, while non-CpG methylation frequency was similar among both cell types. CpG methylation levels accurately distinguished ICM from TE cells epigenetically. Conclusion(s) We describe an effective workflow for isolating and sequencing single ICM and TE cells from human blastocysts. The use of methylation profiling can help distinguish these two cell populations better then morphologic identification alone. TE cells had significantly lower levels of DNA methylation, which may be explained in part by the fact that these cells have begun the process of differentiation and are transcriptionally more active than ICM. This approach may be used to explore the genetic complexities within human embryos, specifically among the two primary cell types seen at this stage of development.

Cover cropping is commonly acknowledged to promote soil health in agriculture. However, contradictory findings on the benefits of cover crops for soil health, crop productivity, economic and ecological factors, as well as the influence of inherent soil parameters on such benefits exist in the scientific literature. Here, we critically assessed evidence of cover crop benefits through a systematic review of the published literature. To access relevant papers, we searched the literature for cover crops and soil health indicators using Scopus (1996-2020), ScienceDirect (1996-2020) and Google scholar (1970-1996) with specific keywords and combinations. Only English research papers including experimental plots and control groups were considered. We analyzed 102 unique peer-reviewed papers and 1494 corresponding unique plots encompassing various cover crops, soil textures, climates, management systems and experimental duration (1-3 years, 4-6 years, 7-10 years and over 10 years). Strong evidence suggests that cover crops can enhance soil structure and promote soil health by improving soil physical and chemical properties, including saturated hydraulic conductivity (mean net change of 105.6 %), total organic carbon (10.1 %), and total nitrogen (20.2 %). On the other hand, cover crops exhibit weak effects on properties like bulk density and microporosity with fairly low values of net change. In most cases, cover crops increase the soil carbon content, including microbial biomass carbon (19.5 %) and particulate organic carbon (49.5 %). In this systematic review, we found limited studies on the effect of cover crops on soil health as influenced by soil texture, regional climate, rainfall and duration of the cover crop practices. The paucity of long-term regional systematic research of soil physics, chemistry and biology makes it difficult to forecast future implications of cover crops on soil health indicators.

Periodontitis is a dysbiotic infectious disease that leads to the destruction of tooth supporting tissues. There is increasing evidence that periodontitis may affect the development and severity of Alzheimer’s disease (AD). However, the mechanism(s) by which periodontal infection impacts the neurodegenerative process in AD remains unclear. In the present study, using an amyloid precursor protein (APP) knock-in (App KI) AD mouse model, we showed that oral infection with Porphyromonas gingivalis (Pg), a keystone pathogen of periodontitis, worsened behavioral and cognitive impairment and accelerated amyloid beta (Aβ) accumulation in AD mice, thus unquestionably and significantly aggravating AD. We also provide new evidence that the neuroinflammatory status established by AD, is greatly complicated by periodontal infection and the consequential entry of Pg into the brain via Aβ-primed microglial activation, and that Pg-induced brain overactivation of complement C1q is critical for periodontitis-associated acceleration of AD progression by amplifying microglial activation, neuroinflammation, and tagging synapses for microglial engulfment. Our study renders support for the importance of periodontal infection in the innate immune regulation of AD and the possibility of targeting microbial etiology and periodontal treatment to ameliorate the clinical manifestation of AD and lower AD prevalence.

Multiple myeloma (MM) is a plasma-cell malignancy that grows in bone marrow (BM). MM resistance to chemotherapy is a major cause of patient mortality. Osteocytes are the most abundant cell type in bone. We have recently demonstrated that MM cells induce osteocyte apoptosis. While the mechanisms responsible for MM-induced osteocyte apoptosis have been described, it is unknown whether apoptotic osteocytes provide feedback that regulates MM cells. To study this, we developed a unique syngenic animal model of murine MM in which osteocyte apoptosis can be induced through targeted expression of human diphtheria toxin receptor (DTR) in osteocytes and subsequent injection of DT. In our current study, we used this DTR mouse model to access the impact of osteocyte apoptosis on MM cell response to the treatments of bortezomib (BTZ) and dexamethasone (DEX), two frontline anti-MM drugs. Eight-week-old DTR mice were i.p. injected with either PBS as control or DT (12.5µg/kg) to induce apoptosis in osteocytes. One week later, DTR+PBS and DTR+DT mice were injected with 106 5TGM1-luc MM cells via tail vein (n=21/group) and after 4 days, randomly assigned to treatment for 4 weeks with PBS or DEX (i.p. 1mg/kg/day) or BTZ (i.p. 0.5mg/kg/3 days) (n=7/group). Bioluminescence imaging and serum IgG2bκ (a soluble marker of 5TGM1-luc MM cells) ELISA, performed after 4-week treatment, demonstrated that osteocyte apoptosis promotes tumor growth in BM; DEX or BTZ treatment significantly inhibited tumor growth in DTR+PBS mice but not in DTR+DT mice. Flow cytometry analysis of 5TGM1-Luc MM cells in the BM further revealed that DEX and BTZ failed to induce MM cell apoptosis/death in DTR+DT mice, in contrast to the effectiveness observed in DTR+PBS mice. Analyses of BM cells harvested from these mice showed that compared with tumor-bearing DTR+PBS+PBS mice, tumor-bearing DTR+DT+PBS mice had significantly higher concentration of immunosuppressive myeloid-derived suppressor cells (MDSCs) and fewer effective CD8+ T cells in the BM. These CD8+ T cells also have a lower expression of the cytotoxic molecules granzyme B and IFN-γ. Importantly, DEX or BTZ treatment improved anti-MM immunity in the BM of tumor-bearing DTR+PBS mice but failed in tumor-bearing OB-Runx2-/- mice. Gemcitabine (GEM) is a FDA-approved anti-cancer agent and MDSC inhibitor used in the treatment of solid tumors but not commonly used in MM therapy. To confirm that MDSCs indeed promote osteocyte-apoptosis-induced MM resistance to DEX/BTZ and to determine if depletion of MDSCs can overcome this resistance, we treated 5TGM1-Luc tumor-bearing DTR+DT mice with PBS, GEM (i.p. 30 mg/kg/week), GEM+DEX or GEM+BTZ for 4 weeks. Bioluminescence imaging and serum IgG2bκ ELISA demonstrated that GEM+DEX or GEM+BTZ reversed MM resistance to DEX and BTZ and resulted in the greatest reduction in tumor burden among all treatments. In conclusion, our findings demonstrate that osteocyte apoptosis promotes MM cell resistance to DEX and BTZ and BM immunosuppression induced by osteocyte apoptosis plays a role in this event. Citation Format: Xiaoxiao Hao, Chao Zhang, Timothy N Trotter, Pramod S Gowda, Yang Yang. Osteocyte apoptosis induces an immunosuppressive microenvironment in bone marrow and promotes myeloma chemoresistance [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy; 2022 Oct 21-24; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2022;10(12 Suppl):Abstract nr A12.

A series of TiZrV0.5Nb0.5Alx refractory high entropy alloys were prepared by arc-metling, aiming to solve the problems of high density, poor oxidation and wear resistance of refractory high entropy alloys at elevated temperature. The microstructure, mechanical and tribological properties were investigated. The results indicate that the increase in Al addition could promote precipitation of Laves phase in the BCC matrix, thus regulation the mechanical properties. The tribology behavior is strongly dependent on the temperature. Below 400 °C, the wear mechanism is dominated by mechanical wear. In this case, the TiZrV0.5Nb0.5Al0.25 alloy with high strength and good ductility at the same time, exhibits optimal wear resistance. When the temperature increases above to 600 °C, the coefficient of friction and the corresponding wear rate dramatically decrease with Al addition, which could be attributed to the preformed formation of dense and stable oxide glazed-layer before wear. Accordingly, the main wear mechanism changes from mechanical wear to oxidation wear. This study provides an effective strategy for improving wear resistance and oxidation resistance of refractory high entropy alloys in a wide temperature range.

The increasing severity of arsenic pollution has progressively threatened human life and attracted much attention. One of the important topics in environmental sciences is to accurately describe the inorganic arsenic transport in heterogeneous porous media, occurring anomalous diffusion phenomenon, which ultimately benefits the control of arsenic pollution. In this paper, we re-evaluate the dataset of the inorganic arsenic transport in porous media in previous work by using a time-Hausdorff fractal model (HADE). Transport experiments of arsenic-carrying (As(V)) ferric humate complex colloids through a quartz sand column were carried out under varying dissolved organic matter (humic acid) concentrations, pH values, ionic strengths, and ferric concentrations. The results show that under our experimental settings, arsenic migration is promoted with the increase of concentrations of HA, ferric ion and sodium ion, and pH to varying degrees. The intensity of arsenic sub-diffusion behavior is opposite to that of arsenic transport. The HADE model can describe the migration behavior of arsenic well, and the value of the time fractal derivative can reflect the diffusion intensity of arsenic migration to a certain extent. By comparing the HADE model, ADE model, and time-fractional model (fADE) to the experimental data, the HADE model can significantly improve all the simulation results of capturing As(V) breakthrough curves (BTCs).

In multiple myeloma, abnormal plasma cells accumulate and proliferate in the bone marrow. Recently, we observed that Runx2, a bone-specific transcription factor, is highly expressed in multiple myeloma cells and is a major driver of multiple myeloma progression in bone. The primary goal of the present study was to identify Runx2-targeting miRNAs that can reduce tumor growth. Expression analysis of a panel of miRNAs in multiple myeloma patient specimens, compared with healthy control specimens, revealed that metastatic multiple myeloma cells express low levels of miR-342 and miR-363 but high levels of Runx2. Reconstituting multiple myeloma cells (CAG) with miR-342 and miR-363 reduced the abundance of Runx2 and the expression of metastasis-promoting Runx2 target genes RANKL and DKK1, and suppressed Runx2 downstream signaling pathways Akt/β-catenin/survivin, which are required for multiple myeloma tumor progression. Intravenous injection of multiple myeloma cells (5TGM1), stably overexpressing miR-342 and miR-363 alone or together, into syngeneic C57Bl/KaLwRij mice resulted in a significant suppression of 5TGM1 cell growth, decreased osteoclasts and increased osteoblasts, and increased antitumor immunity in the bone marrow, compared with mice injected with 5TGM1 cells expressing a miR-Scramble control. In summary, these results demonstrate that enhanced expression of miR-342 and miR-363 in multiple myeloma cells inhibits Runx2 expression and multiple myeloma growth, decreases osteolysis, and enhances antitumor immunity. Thus, restoring the function of Runx2-targeting by miR-342 and miR-363 in multiple myeloma cells may afford a therapeutic benefit by preventing multiple myeloma progression. Implications: miR-342 and miR-363–mediated downregulation of Runx2 expression in multiple myeloma cells prevents multiple myeloma progression. Mol Cancer Res; 16(7); 1138–48. ©2018 AACR.

The international staging system (ISS), based on serum beta-2 microglobulin and albumin, is used to predict survival in multiple myeloma, but its prognostic significance in diffuse large B-cell lymphoma (DLBCL) remains unknown. Herein, we retrospectively analyzed 215 de novo DLBCL patients. According to ISS, there were 90 of 215 (41.9%) patients in stage I, 98 of 215 (45.6%) in stage II and 27 of 215 (12.6%) in stage III group. Patients with ISS stage II/III showed shorter overall survival (OS) and event free survival (EFS) than those with stage I treated with R-CHOP (p = 0.012 and p = 0.043, respectively), but not those treated with CHOP regimen (p > 0.05). Multivariable analysis revealed that ISS, independent of IPI, indicated different survival in both OS (HR, 5.690; 95% CI, 1.270–25.495, p = 0.023) and EFS (HR, 2.116; 95% CI, 1.005–4.455, p = 0.049) in DLBCL patients treated with R-CHOP. ISS could identify patients with better outcome in intermediate-high/high IPI risk patients (p

// Xiaoxiao Hao 1, * , Yongqiang Wei 1, * , Xiaolei Wei 1 , Lizhi Zhou 2 , Qi Wei 1 , Yuankun Zhang 1 , Weimin Huang 1 and Ru Feng 1 1 Department of Hematology, Nanfang Hospital, Southern Medical University, Guangzhou, China 2 Department of Biostatistics, School of Public Health, Southern Medical University, Guangzhou, China * These authors contributed equally to this work Correspondence to: Ru Feng, email: ruth1626@hotmail.com Keywords: diffuse large B-cell lymphoma, glasgow prognostic score, inflammation-based prognostic score Received: March 16, 2017 Accepted: August 26, 2017 Published: September 11, 2017 ABSTRACT Inflammation-based prognostic scores, such as the glasgow prognostic score (GPS), prognostic index (PI), prognostic nutritional index (PNI), neutrophil lymphocyte ratio (NLR) and platelet lymphocyte ratio (PLR) were related to survival in many solid tumors. Recent study showed that GPS can be used to predict outcome in diffuse large B-cell lymphoma (DLBCL). However, other inflammation related scores had not been reported and it also remained unknown which of them was the most useful to evaluate the survival in DLBCLs. In this retrospective study, a number of 252 newly diagnosed and histologically proven DLBCLs from January 2003 to December 2014 were included. The high GPS, high PI, high NLR, high PLR and low PNI were all associated with poor overall survival ( p < 0.05) and event-free survival ( p < 0.05) in univariate analysis. Multivariate analysis indicated that GPS (HR = 1.781, 95% CI = 1.065–2.979, p = 0.028) remained an independent prognostic predictor in DLBCL. The c-index of GPS (0.735, 95% CI = 0.645–0.824) was greater than that of PI (0.710, 95% CI = 0.621–0.799, p = 0.602), PNI (0.600, 95% CI = 0.517–0.683, p = 0.001), PLR (0.599, 95% CI = 0.510–0.689, p = 0.029) and NLR (0.572, 95% CI = 0.503–0.642, p = 0.005) by Harrell’s concordance index. Especially in DLBCLs treated with R-CHOP, GPS still remained the most powerful prognostic score when comparing with others ( p = 0.001 and p < 0.001, respectively for OS and EFS). In conclusion, it is indicated that inflammation-based prognostic scores such as GPS, PI, NLR, PNI and PLR all could be used to predict the outcome of DLBCLs. Among them, GPS is the most powerful indicator in predicting survival in DLBCLs, even in the rituximab era.

Anomalous relaxation and diffusion processes have been widely quantified by fractional derivative models, where the definition of the fractional-order derivative remains a historical debate due to its limitation in describing different kinds of non-exponential decays (e.g. stretched exponential decay). Meanwhile, many efforts by mathematicians and engineers have been made to overcome the singularity of power function kernel in its definition. This study first explores physical properties of relaxation and diffusion models where the temporal derivative was defined recently using an exponential kernel. Analytical analysis shows that the Caputo type derivative model with an exponential kernel cannot characterize non-exponential dynamics well-documented in anomalous relaxation and diffusion. A legitimate extension of the previous derivative is then proposed by replacing the exponential kernel with a stretched exponential kernel. Numerical tests show that the Caputo type derivative model with the stretched exponential kernel can describe a much wider range of anomalous diffusion than the exponential kernel, implying the potential applicability of the new derivative in quantifying real-world, anomalous relaxation and diffusion processes.

DNA Mismatch repair (MMR) deficiency is a major cause of hereditary non-polyposis colorectal cancer, and is also associated with increased risk of several other cancers. This is generally ascribed to the role of MMR in avoiding mutations by correcting DNA replication errors. In MMR knockout mice very high frequencies of somatic mutations, up until 100-fold of background, have been reported. However, these results have been obtained using bacterial reporter transgenes, which are not representative for the genome overall, and mutational patterns of MMR deficiency remain largely unknown. To fill this knowledge gap, we performed single-cell whole-genome sequencing of lung fibroblasts of Msh2−/− and wild-type mice. We observed a 4-fold increase of somatic single nucleotide variants (SNVs) in the fibroblasts of Msh2−/− mice compared to those of wild-type mice. The SNV signature of Msh2 deficiency was found to be driven by C>T and T>C transitions. By comparing it to human cancer signatures, we not only confirmed the inferred MMR-deficiency-related etiology of several cancer signatures but also suggested that MMR deficiency is likely the cause of a cancer signature with its etiology previously unknown. We also observed a 7-fold increase of somatic small insertions and deletions (INDELs) in the Msh2−/− mice. An elevated INDEL frequency has also been found in human MMR-related cancers. INDELs and SNVs distributed differently across genomic features in the Msh2−/− and control cells, with evidence of selection pressure and repair preference. These results provide insights into the landscape of somatic mutations in normal somatic cells caused by MMR deficiency.SignificanceOur results show that MMR deficiency in the mouse is associated with a much lower elevation of somatic mutation rates than previously reported and provides the first MMR whole-genome mutational landscapes in normal somatic cells in vivo.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.

Activation of retrotransposons and their insertions into new genomic locations, i.e., retrotranspositions (RTs), have been identified in about 50% of tumors. However, the landscape of RTs in different, normal somatic cell types in humans remains largely unknown. Using single-cell whole-genome sequencing we identified 528 RT events, including LINE-1 (L1), and Alu, in 164 single cells and clones of fibroblasts, neurons, B lymphocytes, hepatocytes and liver stem cells, of 29 healthy human subjects aged from 0 to 106 years. The frequency of RTs was found to vary from

This paper proposes a new hybrid modulation mode (HMM) to eliminate the zero-crossing distortion of grid current and enable reactive power provision for a H6 configuration PV (photovoltaic) grid-tied inverter. The common mode voltage, leakage current, and efficiency for the proposed approach are also analyzed. In order to improve grid frequency tracking a novel frequency self-adaptive proportional-integral-resonant (FSAPIR) controller is implemented which reduces error for changes in grid frequency. The proposed approach provides the basis for accurately adjusting the active and reactive current without error to improve the grid support capability of the inverter. Theoretical analysis, simulation, and experiment verify the newly proposed modulation mode and controller.

Agriculture, a strategic industry that secures the country and stabilizes society, has invariably been valued by China to feed its 1.4 billion people. However, the long-term extensive development of agriculture has led to a series of water problems such as groundwater overexploitation, water shortages, and water pollution in some areas of China, resulting in great pressure on the ecological environment and social economy. In this paper, we constructed a dynamic optimization simulation model based on the input–output (I–O) approach within the horizon of 2012–2025 to explore the synthetic optimal policies for achieving both socioeconomic development, and water resources and water environment protection in a node city on the middle route of the South-to-North Water Transfer Project, Handan. Six scenarios based on various synthetic policies were adopted. The simulation results show that during the simulation period, the maximum development of the social economy can be achieved under the premise of achieving the specific objectives of water resources and water environment protection in a comprehensive scenario. In addition, the water environment improved: the annual chemical oxygen demand (COD) emissions reduced by 7.5%, 10.3%, 5.0%, and 5.9% in pig farming, cattle farming, chicken farming, and households, respectively. Furthermore, the water supply–demand structure was optimized. The water demand of wheat planting and groundwater extraction decreased at an annual rate of 9.1% and 7.7%, respectively. The supply of reclaimed water increased at an annual rate of 9.2%. The model adapted in the paper can simulate and evaluate the impact of all the feasible synthetic policies on the economy, water resources, and water environment, and provide targeted, scientific recommendations for solving the agricultural water use problems in other regions.

Both process changes and sensor faults can result in the unexpected deviation of the sensor reading from its anticipated values. When the anomaly of sensor measurement occurs, in order to take appropriate actions to diminish or eliminate related dangerous factors, identifying the source of abnormal conditions is a very crucial step. To reach that, an abnormal condition diagnosis method based on virtual sensing technology is proposed. Once the abnormal situation of sensor measurement is detected, the virtual sensor is constructed based on the Gauss Process Regression and Moving Windows algorithm. Through analyzing prediction residual, the task of differentiating between sensor and system faults can be performed. In order to verify the performance of this proposal, a case study is carried out on CSTH platform, where both scenarios of sensor faults and process changes are considered. The simulation results show the effectiveness of this method.

The germline mutation rate has been extensively studied and has been found to vary greatly between species, but much less is known about the somatic mutation rate in multicellular organisms, which remains very difficult to determine. Here, we present data on somatic mutation rates in mice and humans, obtained by sequencing single cells and clones derived from primary fibroblasts, which allows us to make the first direct comparison with germline mutation rates in these two species. The results indicate that the somatic mutation rate is almost two orders of magnitude higher than the germline mutation rate and that both mutation rates are significantly higher in mice than in humans. Our findings demonstrate both the privileged status of germline genome integrity and species-specific differences in genome maintenance., Germline mutation rates are known to vary between species but somatic mutation rates are less well understood. Here the authors compare mice and humans, observing that somatic mutation rates were nearly two orders of magnitude higher in both species, with both mutation rates significantly higher in mice.

The binding of quercetin to lysozyme (LYSO) in aqueous solution was investigated by fluorescence spectroscopy, UV-vis absorption spectroscopy and molecular simulation at pH 7.4. The fluorescence quenching of LYSO by addition of quercetin is due to static quenching, the binding constants, K ( a ), were 3.63 × 10(4), 3.31 × 10(4) and 2.85 × 10(4) L·mol(-1) at 288, 298 and 308 K, respectively. The thermodynamic parameters, enthalpy change, ∆H, and entropy change, ∆S, were noted to be -7.56 kJ·mol(-1) and 61.07 J·mol(-1)·K(-1). The results indicated that hydrophobic interaction may play a major role in the binding process. The distance r between the donor (LYSO) and acceptor (quercetin) was determined as 3.34 nm by the fluorescence resonance energy transfer. The synchronous fluorescence spectroscopy showed the polarity around the tryptophan residues increased and the hydrophobicity decreased. Furthermore, the study of molecular simulation indicated that quercetin could bind to the active site (a pocket made up of 24 amino-acid residues) of LYSO mainly via hydrophobic interactions and that there were hydrogen interactions between the residues (Gln 57, Ile 98) of LYSO and quercetin. The accessible surface area (ASA) calculation verified the important roles of tryptophan (Trp) residues during the binding process.

The binding mechanism of the interactions of halide ions (F–, Br– and I–) with bovine serum albumin (BSA) and hemoglobin (Hb) were studied at different temperatures, by using ion-selective electrodes. The experimental data were treated according to Klotz equation, and the number of binding sites and the binding constants were determined. The results show that the binding sites of F– on protein molecules are more than those of Br– and I–. Additionally, the number of the binding sites for halide ions on protein molecules increases with increasing temperature. This study also indicates that the binding constants for the interactions of halide ions with proteins gradually decrease as the size of halide ions and temperature increases. These behaviors were reasonably interpreted with the structural and thermodynamic factors. The thermodynamic functions at different temperatures were calculated with thermodynamic equations, and the enthalpy change for the interactions were also determined by isothermal titration calorimetry (ITC) at 298.15 K, which indicate that the interactions of halide ions with proteins are mainly electrostatic interaction.

The prognostic value of CD30 expression in diffuse large B-cell lymphoma (DLBCL)remains controversial. Herein, we performed this retrospective study to investigate the clinical and prognostic significance of CD30 expression in patients with DLBCL.Among all the 146 patients, the expression of CD30 was observed in 23 cases (15.7%).The DLBCL patients with CD30 expression showed more likely to present B symptoms, bone marrow involvement, non-germinal centre B-cell-like (Non-GCB) DLBCL, BCL-2 and Ki-67 overexpression (p

The peripheral blood lymphocyte to monocyte ratio (LMR) at diagnosis has been used to predict survival in diffuse large B-cell lymphoma (DLBCL) patients, but its prognostic significance with respect to different cell-of-origin (COO) subtypes remains unknown. We retrospectively analyzed 168 de novo DLBCL patients in this study and found that a low LMR (≤2.6) correlates with B symptoms, elevated LDH, advanced Ann Arbor stage and higher international prognostic index (IPI) score (p

Objective: To observe the influence of biological character of acute lymphoblastic leukemia cell line Jurkat in vitro and in vivo after overexpression of C/EBPa and PU.1. Methods: To establish transducing plasmids, using 293T cells to package lentiviral vector and to transfect Jurkat cell line. Flow cytometry is used to analyze the expression of membrane cluster differentiation antigen, apoptosis rate, proliferation and cell cycle of cells. Cell morphology and Q-PCR was used to analyze the changes of cell morphology and molecule gene. Six to eight weeks old immunodeficiency (NSI) mice were transplanted by tail vein injected with 2x106 cells after transduction. To observe the biological behavior and survival time, using Flow cytometric, cell morphology, pathological HE staining, immunohistochemistry to analyze the invasion of leukemia cells. Results: The expression of CD3 (mark antigen of lymphocyte) fall as low as 20% after overexpression of C/EBPa and PU.1, CD11b (mark antigen of myeloid cell) increase as high as 30%, which has significant difference with Jurkat-gfp group. The apoptosis rate has significantly increase, proliferation rate has significantly decrease after overexpression of C/EBPa, which blocked in S period. Adding myeloid cytokine (GM-CSF, M-CSF, IL-3, FLT-3L,100ng/mL) on the supernatant of transduction cells, Jurkat cells can be reprogrammed to attached cells from suspension after overexpression of C/EBPa and PU.1 in the 5th day. Attached cells can chemotaxis and consuming yeast. Cell multicore and small amount of purple particles in cytoplasm has observed in Jurkat after overexpression of C/EBPa and PU.1, high expression of myeloid gene (CD14, CD64) and low expression of lymphocyte gene (BCL-11B, GATA-3) has observed in cells after transduction. All the mice have leukemia between 1 to 2 months after injection with transfected cells. The survival time is longest in overexpression of C/EBPa group, which shows significant difference (P Conclusion: Jurkat cells can be partly reprogrammed to monocyte- macrophages, while with the help of myeloid cytokines, it can be reprogrammed to macrophages with chemotactic and phagocytic function. Jurkat cells can be partly reprogrammed to monocyte- macrophages in vivo and prolong the survival time after overexpression of C/EBPa and PU.1. Disclosures No relevant conflicts of interest to declare.

Objective International prognostic index (IPI) has been widely used for predicting outcome in diffuse large B cell lymphoma. Although the introduction of rituximab to chemotherapy has dramatically improved the outcome of DLBCL, it also changed the prognostic value of IPI. The National Comprehensive Cancer Network IPI (NCCN-IPI) and age-adjusted IPI (aaIPI) were used to evaluate the prognosis for DLBCL in the rituximab era. However which one of them is more powerful in predicting survival remains unknown in Chinese patients. Patients and Methods A total of 334 patients with de novo diffuse large B-cell lymphoma diagnosed from 2003 to 2012 were included. All patients were treated with CHOP with or without rituximab. They were divided into CHOP and R-CHOP groups. IPI, NCCN-IPI and aaIPI score were recorded. Survival was performed according to the Kaplan-Meier (K-M) curves using the log-rank test. The predictive abilities of PI, NCCN-IPI and aaIPI were investigated by Harrell's C-statistics. Result Compared with the IPI, the NCCN-IPI and the aaIPI had better discrimination in different scores in all patients treated with or without rituximab. Both the NCCN-IPI and aaIPI had power in discriminating low, low-intermediate and high-intermediate risk groups. In comparison, the NCCN-IPI discriminated low-intermediate and high-intermediate risk groups (5-year overall survival [OS]: 74% vs 50%) better than the aaIPI (5-year OS: 80% vs 62%) in all patients and in the R-CHOP group with the NCCN-IPI 5-year OS: 85% vs 67% while the aaIPI: 87% vs 77%. In the CHOP group, the aaIPI discriminated low-intermediate and high-intermediate risk groups better (5-year OS: 70% vs 25%) than the NCCN-IPI (5-year OS: 60% vs 24%). According to the Harrel C statistic, the NCCN-IPI showed higher discrimination of the different scores (C_index: 0.669 ) than the aaIPI (C_index: 0.663 ) in all the patients (C_index: 0.669 vs. 0.663), especially in the R-CHOP group (C_index: 0.681 vs. 0.636). In the CHOP group, the aaIPI had stronger power (C_index: 0.711 ) than the NCCN-IPI (C_index: 0.683 ) in discriminating different scores. Conclusion The NCCN-IPI is more powerful than the IPI for predicting survival in the rituximab era. In patients aged 60 or younger and treated with CHOP without rituximab, aaIPI is a preferable tool for evaluating prognosis. Disclosures No relevant conflicts of interest to declare.

Inflammation-based prognostic scores, such as the glasgow prognostic score (GPS), prognostic index(PI), prognostic nutritional index(PNI), neutrophil lymphocyte ratio(NLR), platelet lymphocyte ratio(PLR) was related to survival in many solid tumors. Recent study showed that GPS can be used to predict outcome in diffuse large B-cell lymphoma(DLBCL). However other inflammation related scores had not been reported in DLBCL, and it also remained unknown which of them was more useful to evaluate the survival in DLBCL. In this retrospective study, a number of 252 newly diagnosed and histologically proven DLBCL patients from January 2003 to December 2014 were included. An elevated GPS, PI, NLR, PNI and PLR were all associated with decreased overall survival(p=0.000, p=0.000, p=0.006, p=0.001 and p=0.001, respectively) and event-free survival (p=0.000, p=0.000, p=0.011, p=0.001 and p=0.009, respectively) in univariate analysis. Multivariate analysis indicated that GPS(RR=1.768, 95%CI=1.043-3.000, p =0.034) remained an independent prognostic predictor in DLBCL. The area under the curve of GPS (0.735, 95%CI=0.645-0.824) was greater than that of PI(0.710, 95%CI=0.621-0.799), PNI(0.600, 95%CI=0.517-0.683), NLR(0.572, 95%CI=0.503-0.642), and PLR(0.599, 95%CI=0.510-0.689) by Harrell's C-statistics. Especially in the DLBCL patients treated with R-CHOP, GPS also remained the most powerful inflammation-based prognostic score when comparing with PI, NLR, PNI and PLR (p=0.004, p=0.000, respectively for OS and EFS). In conclusion, these results indicate that Inflammation-based prognostic scores such as GPS, PI, NLR, PNI and PLR can be used to evaluate the outcome in DLBCL patients. Among them, GPS is the most powerful tool in predicting survival in DLBCL patients, even in the rituximab era. Disclosures No relevant conflicts of interest to declare.

Tumor-associated macrophages (TAMs), as the major component in tumor microenvironment, have been reported to correlate with the prognosis in diffuse large B-cell lymphoma (DLBCL). However due to the different subtypes of TAMs and the given therapy regimen, the prognostic value of TAMs in DLBCL patients remains controversial. To explore the prognostic significance of different TAMs subtypes in DLBCL patients treated with R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone), we retrospectively analyzed 77 de novo DLBCL patients in present study. TAM markers including CD68 and CD163 were evaluated by immunohistochemistry. The cutoff value of CD68, CD163 and CD163/CD68 ratio was determined by receiver operating characteristic curve. In a total of 77 patients, CD68 high expression was found in71.4 %( 55/77) DLBCL patients and CD163 high expression in 51.4 %( 40/77 patients. Patients with high CD163 expression were more present with extranodal sites (P =0.049). However other clinical parameters including age, gender, Ann Arbor stage, performance stage, B symptoms, LDH and international prognostic index (IPI) showed no difference in patients with high and low CD68 or CD163 expression (P >0.05). CD68 expression has less effect on overall survival (OS) and event-free survival (EFS) (P =0.852, 0.782). However, high CD163 expression showed poor OS and EFS in DLBCL patients (P =0.006, 0.004) and also high CD163/CD68 ratio (P Disclosures No relevant conflicts of interest to declare.

Introduction: Pegaspargase(PEG-ASP) and L-asparaginase(L-ASP) has been widely used in the treatment of acute lymphatic leukemia, changes of coagulation function after treatment are not actually the same. So we performed the present study to analyze coagulation disorder after PEG-ASP and L-ASP treatment of adults with acute lymphatic leukemia. Methods and materials: Totally 153 hospitalized patients with acute lymphatic leukemia treated with L-ASP or PEG-ASP were studied from January, 2010 to January, 2015. Of all 153 patients analyzed, they received L-ASP treatment 158 dose times and PEG-ASP treatment 60 dose times respectively. Results: There is no difference of the distribution of age, sex, white blood cell count at diagnosis and risk factors of the disease. Agranulocytosis combining with intestinal infection, bleeding or thrombosis and the inducing remission rate between the two groups has no significant difference (p=0.11,0.61,0.33). The total infusion of fresh frozen plasma or cryoprecipitate or fibrinogen after treatment shows no significant difference between the two groups (p=0.11,0.75,0.21).Fibrinogen level decreases slower in the treatment of PEG-ASP(9.37 day vs 7.40 day, p=0.02) than that of L-ASP. What's more, fibrinogen decreases slower when L-ASP used at interval compared with continuous use, however, the incidence rate of bleeding and related complications is higher when used at interval early (p =0.028). Conslusion: Because of the preponderance to monitor the changes of fibrinogen and the equal rate of complications and inducing remission rate, it is recommended to use PEG-ASP. L-ASP used at interval can monitor the coagulation function easily than continuous use, but the early use of L-ASP may overlay the drug side effects and related hematology toxicity caused by chemotherapy, then cause an increased incidence of complications. Disclosures No relevant conflicts of interest to declare.

The binding properties of amino phosphate ester derivatives, compound 1 and 2 with calf thymus DNA (CT-DNA) were investigated by UV spectra, fluorescence spectra, molecular modeling and isothermal titration calorimetry (ITC). The intrinsic binding constants K b of compound 1 and 2 with CT-DNA were determined by fluorescence spectroscopy and ITC, respectively. The results indicated that the two compounds bind to CT-DNA with different binding affinity, which is in the order of compound 1 > compound 2. At the same time, fluorescence spectra suggested that the mechanism of the binding of the two compounds to CT-DNA is a static enhancing type. According to the ethidium bromide displacement experiments, UV spectra, molecular modeling and ITC studies, it can be concluded that compound 1 and 2 are intercalators that can slide into the G–C rich region of CT-DNA. Furthermore, ITC data showed that compound/DNA binding is enthalpy controlled.

Background Diffuse large B-cell lymphoma (DLBCL) is the most common non-Hodgkin lymphoma (NHL). Although international prognostic index (IPI) has been widely used in DLBCL to evaluate the prognosis, there is need to find new biomarkers to predict outcome. Tumor microenvironment has been reported to be associated with both metastatic spread and diminished in some tumors. Tumor-associated macrophages (TAMs) and nature-killer (NK) cells play an important role in the tumor microenvironment. Therefore, we performed this study to investigate the prognostic implications of TAMs including M1 as well as M2 and NK cells in DLBCLs. Materials and Methods We reviewed 77 cases of DLBCL diagnosed between June 2004 and October 2012, who treated with rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone (R-CHOP). Patients were divided into GCB and non-GCB subgroups by Hans’ algorithm. We performed immunohistochemical stains using CD68, CD163 and CD57 to mark M1 macrophages, M2 macrophages and NK cells on paraffin tissue section. The percentage of TAMs was recorded and graded as: 0 if less than 5%, 1 if 5-25%, and 2 if greater than 25% of the total cells present in tumor cells rich nodules were positive for CD68 or CD163. And counting the mean number of the CD57+ cells in 5 fields under high-power magnification (400 times): 0 if less than 5 positive cellsG1 if more than 5 positive cells. Results Of the 77 DLBCL patients treated with R-CHOP, there were 51 males (66.2%), 26 females (33.8%). The median age was 52 years (range, 16-81 years). The median follow-up was 38.1 months. The median percentage for CD68 was 12.8 %( range, 2.4–50.84), for CD163 was 12.3 %( range, 1.0–39.2). The median counts of CD57 cells were 5 (range, 1.0-78) per unit area. In univariate analysis, an increase in CD68-positive cells was related to improved EFS (grade 0 vs. grade 2, p = 0.036). By contrast, an increased number of CD163-positive cells were significantly poor associated with OS, (grade0 vs grade2, p = 0.033). Patients with elevated NK cells tended to have no effect on prognosis. In multivariate analysis, an increased CD68 -positive cells was an independent predictors of EFS (HR 0.467, 95%CI 0.236-0.922, P=0.028), an decreased CD163-positive cells and lower LDH were independent predictors for OS (HR 2.287, 95%CI 1.043-5.013, P=0.039). Conclusion These results suggest that M1 macrophages might predict favorable clinical outcome and M2 macrophages predict poor clinical outcome. However, the NK cells have no prognostic significance in DLBCL patients treated with RCHOP. Disclosures No relevant conflicts of interest to declare.