Double Fusion Research Articles

Three-Dimensional Reconstruction of Dilute Bubbly Flow Field With Light-Field Images Based on Deep Learning Method

The three-dimensional reconstruction of bubble flow field is of great significance to study the motion of gas-liquid two phase flow. Combined with the abundant information of bubbles in the light field images, a fast and accurate 3D reconstruction method for dilute bubbly flow based on the deep learning algorithm DIF-LeNet (Double Information Fusion with LeNet5 net) is proposed in this work. The calibration method and bubble segmentation algorithm of light field image for data processing is detailed. DIF-LeNet realizes the fusion of different dimension data and regression prediction. By DIF-LeNet, bubble depth could be predicted by fusing the refocused bubble image with the focal distance of the refocused image. Then, the three-dimensional bubble field could be reconstructed with the bubble depth, bubble center and bubble diameter. Comparing with the reconstruction method based on statistics and LeNet5, the reconstruction accuracy and speed of the proposed method based on DIF-LeNet are improved. Especially for the bubble without focused image in the refocused image sequence, the effect of 3D reconstruction based on DIF-LeNet is much prominent.

NRG1 fusions in breast cancer

BackgroundNRG1 gene fusions may be clinically actionable, since cancers carrying the fusion transcripts can be sensitive to tyrosine kinase inhibitors. The NRG1 gene encodes ligands for the HER2(ERBB2)-ERBB3 heterodimeric receptor tyrosine kinase, and the gene fusions are thought to lead to autocrine stimulation of the receptor. The NRG1 fusion expressed in the breast cancer cell line MDA-MB-175 serves as a model example of such fusions, showing the proposed autocrine loop and exceptional drug sensitivity. However, its structure has not been properly characterised, its oncogenic activity has not been fully explained, and there is limited data on such fusions in breast cancer.MethodsWe analysed genomic rearrangements and transcripts of NRG1 in MDA-MB-175 and a panel of 571 breast cancers.ResultsWe found that the MDA-MB-175 fusion—originally reported as a DOC4(TENM4)-NRG1 fusion, lacking the cytoplasmic tail of NRG1—is in reality a double fusion, PPP6R3-TENM4-NRG1, producing multiple transcripts, some of which include the cytoplasmic tail. We hypothesise that many NRG1 fusions may be oncogenic not for lacking the cytoplasmic domain but because they do not encode NRG1’s nuclear-localised form. The fusion in MDA-MB-175 is the result of a very complex genomic rearrangement, which we partially characterised, that creates additional expressed gene fusions, RSF1-TENM4, TPCN2-RSF1, and MRPL48-GAB2.We searched for NRG1 rearrangements in 571 breast cancers subjected to genome sequencing and transcriptome sequencing and found four cases (0.7%) with fusions, WRN-NRG1, FAM91A1-NRG1, ARHGEF39-NRG1, and ZNF704-NRG1, all splicing into NRG1 at the same exon as in MDA-MB-175. However, the WRN-NRG1 and ARHGEF39-NRG1 fusions were out of frame. We identified rearrangements of NRG1 in many more (8% of) cases that seemed more likely to inactivate than to create activating fusions, or whose outcome could not be predicted because they were complex, or both. This is not surprising because NRG1 can be pro-apoptotic and is inactivated in some breast cancers.ConclusionsOur results highlight the complexity of rearrangements of NRG1 in breast cancers and confirm that some do not activate but inactivate. Careful interpretation of NRG1 rearrangements will therefore be necessary for appropriate patient management.

“Three-Education” mode of Civil Engineering talents Based on “Double Chain Fusion”

The core of developing application-oriented undergraduate education is to create a first-rate ecology that is most conducive to the cultivation of undergraduate talents. Based on the education chain and industrial chain, the civil engineering talent cultivation mode of moral education, practical education and innovative education is constructed. The national excellent party building model branch has been established, and the “model worker” training studio has been set up to play the leading role of party building education. Engineering experience, scientific research experience and other practical education methods are explored. Innovation education mechanism of teaching practice, innovation activities, and science and technology service has been constructed. The results show that the model enables students to succeed, enterprises to improve economic efficiency and market competitiveness, and teachers to benefit a lot.

A Patient with Chronic Lymphocytic Leukemia, Chronic Myeloid Leukemia and Multiple Myeloma

Chronic lymphocytic leukemia (CLL) is an indolent lymphoproliferative disorder and is manifested by progressive accumulation of B cells in the blood, bone marrow and lymphatic tissues. Chronic Myeloid Leukemia (CML) is a clonal myeloproliferative disorder characterized by the presence of all the stages of myeloid development in the peripheral blood, and it is believed to be driven by the aberrant protein tyrosine kinase, a product of the mutant BCR-ABL1 gene.Multiple Myeloma (MM) is characterized by the accumulation of clonal plasmcells in the bone marrow with skeletal lesions, anemia, hypercalcemia and renal failure. Our patient is a 78 year-old man. In 2014 diagnosis of CLL and monoclonal gammopathy of undetermined significance (MGUS).At diagnosis: HB 13.5 g/dl; normal renal function;calcium 8.7 mg/dl;IgG 1678 mg/dl,serum immunoelectrophoresis: IgG kappa, Bence Jones kappa; total protein 7.5 g/dl, beta1 6,5%, beta2 24,1%;peripheral blood immunophenotyping showed CLL, FISH:negative;Cariotype: 46, XY; RX skeleton: positive for osteolytic lesions, total body TC scan: adenopathies of 18 mm and 15 mm at bilateral axillary level, norma spleen, adenopathy of 22 cm in the left obturator iliac region; presence of left hip prosthesis; bone marrow biopsy: localization by low-grade plasmacytoma.No CLL.The patient was only observed until April 2015, when there was a presence of myelocytes and metamyelocytes in peripheral blood and an increased spleen (18 cm). So he performed : bone marrow aspirate: diagnosis of CML (Sokal Score: 1,34 H; Eutos Score: 60 L, Hasford Score 1488,5);bone marrow biopsy: suggestive for a myeloproliferative disease (CML), MGUS with a modest lymphoid B component,BCR-ABL: 60;FISH: pathological presence of double fusion signal of the ABL1 and BCR loci in 209 of 271 interphase nuclei examined (77%).The patient started therapy with Imatinib, 400mg/die until July 2015, on the basis of the good response to treatment and the progressive increase of the M component that confirmed the progression to MM: Hb 9.1 g/dL, creatinine 1,1 mg/dl;calcium 10,5 mg/dl;total protein: 8,6 g/dl, gamma 48.02% (CM 4 gr); IgG 3536 mg/dl, cariotype: male with t (9; 22) and Philadelphia chromosome (25%);BCR-ABL: 14,32; bone marrow aspirate: plasmacells 15%;bone marrow biopsy: intermediate-interstitial plasmacytoma, CLL / lymphoma; RMN whole body: hyperintensity at the level of the seventh right rib; PET: osteolytic lesions of the side arch tenth right rib, right iliac bone, left iliac region, right tibia third diaphyseal.RX right hemithorax: osteolytic area at the level of the seventh right rib. So the patient started treatment with Bortezomib, Desamethasone, Alkeran (total 7 cycles).On March 2016, he performed a radiography that showed many osteolytic areas of 45 mm on third distal femur, third proximal and intermediate tibia, third proximal and third distal of fibula.A second PET documented a further MM progression due to new bone localizations and a left tibia biopsy showed localization disease. Radiotherapy colleagues have ruled out the usefulness of a radiation therapy program in consideration of the cerebral damage risk. On June 2016 the patient started a treatment with Lenalidomide for 15 days, interspersed by Glivec, maintaining the disease stable. In September 2017 he developed diplopia and with a nasal surgery, only inflammatory tissue was exported. A revision of the material confirmed plasmacytoma localization. In the same period appearance of a right gluteus sore treated initially with surgical dressing.As blood tests revealed increase of paraprotein levels, bone marrow biopsy resulted negative to myeloma and lymphoma diseases, instead a gluteal skin biopsy revealed plasmacytoma. It was decided to treat cerebral localization due to diplopia and peripherical paralysis. Radiotherapy was started on April 2018 (18 sessions). Bone marrow aspirate test showed plasmacells 15%, BCR-ABL dosage: 213,87, M component increase(5gr), IgG 4440 mg/dl, creatinine and serum calcium: normal. Due to disease progression, a rescue chemotherapy was started according to PAD protocol. After 4 cycles, a bone marrow aspirate documented the presence of plasmacells equal to 80%.The cytogenetic study confirmed the presence of a complex karyotype. So the patient started therapy with Daratumumab, Lenalidomide and Desamethasone which is currently ongoing with an excellent hematological and clinical response Disclosures Ciolli: Janssen: Honoraria; Abbvie: Research Funding.

Selective cement augmentation of cranial and caudal pedicle screws provides comparable stability to augmentation on all segments in the osteoporotic spine: a finite element analysis.

BackgroundCement-augmented pedicle screw instrumentation (CAPSI) has been proven to significantly increase the biomechanical stability in the osteoporotic lumbar spine. However, besides the merits, it is responsible for the inevitable cement leakage growing with more instrumented segments and volumes involved. This study aimed to compare the biomechanical performance of pedicle screws augmented on all segments with those augmented only on the cranial and caudal vertebrae selectively.MethodsThe finite element model of L3-S1 was modeled with the CT data of a healthy volunteer, the solid/fenestrated pedicle screws from micro-CT scans of physical screws, and bone cement from the CT scans of a postoperative patient with CAPSI. Three different augmented strategies for pedicle screws were taken into consideration: augmentation at each pedicle trajectory (Model A), selective augmentation at the cranial and caudal pedicle trajectories (Model B), and pedicle trajectories without augmentation (Model C). A total of six surgical models were constructed: Models A, B, and C were subdivided into double segmental fusion from L4 to S1 (Models A1, B1, and C1) and multi-segment fusion from L3 to S1 (Models A2, B2, and C2). The Range of motion (ROM), stress on the cage, and stress on the fixed segments were compared among the six models.ResultsThe ROM at the fusion segments decreased in all instrumentation models. The ROMs of Model B and Model A are similar in each direction, while that of Model C is significantly larger. The differences in the ROMs between Model A and Model B were noted to be less than 0.1°. Compared with Models A1 and A2, the peak Von Mise stress on the cage-endplate interface and pedicle screws were slightly higher in Models B1 and B2. In contrast, the stress of Models C1 and C2 increased significantly. The compressive stress was concentrated in the screw head, the cranial and caudal screws, and rods.ConclusionsThe selective augmentation of pedicle screws is capable of providing reliable stability in short-segment posterior fixation (2- or 3-level). It could be a potential optimal procedure to minimize the associated complications of CAPSI.

A visible-infrared double band photodetector absorber

In this paper, a double-band visible-infrared photodetector absorber is proposed, which consists of a modified microbolometer and a metallic reflective layer which has a deep well. The simulated results show that the absorber proposed has high absorption peaks both at visible band (400–800 nm) and infrared band (8–12 μm), and the absorptivity at infrared band is more than 85% while the absorption rate at visible band exceeds 70%. By analyzing the simulation results, the absorption mechanism is discussed. The designed absorber not only has high absorption rate, but also realizes visible-infrared double band coaxial absorption with single pixel integration, hence it is favorable for various applications, such as night and day vision coaxial detecting, double band fusion detecting, and optical filter and bio-chemical sensor.

Secure Storage Method of Ship Communication Data Based on Double Key and Hadoop

Yang, H.; Zhang, C., and Zhao, B., 2020. Secure storage method of ship communication data based on double key and Hadoop. In: Yang, Y.; Mi, C.; Zhao, L., and Lam, S. (eds.), Global Topics and New Trends in Coastal Research: Port, Coastal and Ocean Engineering. Journal of Coastal Research, Special Issue No. 103, pp. 952–955. Coconut Creek (Florida), ISSN 0749-0208.In order to improve the security storage and transmission ability of ship communication data, it is necessary to dynamically encrypt the ship communication data, and a dynamic encryption method of ship communication data based on double key and Hadoop is proposed. The secure key transmission model of ship communication data is constructed, the multidimensional mapping structure of ship communication data is reorganized by the method of spatial quantization coding, the encryption protocol of double key automatic control of ship communication data is established, the double key Hadoop cloud platform coding configuration method is adopted, the double key random quantization coding and feature fusion processing of ship communication data are carried out, the encryption key and decryption key are constructed, and the dynamic encryption design of ship communication data is carried out according to the results of double key encryption and Hadoop cloud platform storage. The simulation results show that the security of dynamic encryption of ship communication data using this method is better, and the fidelity of ship communication data transmission is higher, which improves the security storage ability of ship communication data and the ability of resisting attack.

Different interbody fustion cages and combined fixation through intermuscular approach for lumbar diseases:a case control study

To explore the clinical effect of combined fixation and interbody bone grafting through intermuscular approach with different fusion cages in the treatment of single segment lumbar diseases. From June 2014 to December 2016, the clinical data of 123 cases of single segment lumbar diseases were analyzed retrospectively, including 44 males and 79 females, aged 22 to 60 years old, with the diseases course of 6 to 84 months. The disease types involved lumbar disc degeneration in 65 cases, lumbar spinal stenosis in 30 cases, MeyerdingⅠslip in 21 cases, giant lumbar disc herniation in 7 cases. Lesions was L3, 4 in 5 cases, L4, 5 in 101 cases, L5S1 in 17 cases. According to the application of different interbody fusion cage, patients were divided into single common cage group, double common cage group and banana type cage group. The operation time, intraoperative hemorrhage, postoperative incision drainage fluid and incision length were observed in three groups; VAS score of lumbar incision and JOA score of preoperative and final follow up were recorded 72 hours after operation;the intervertebral space height, Cobb angle of lumbar coronal and sagittal plane before and after operation, and interbody fusion 12 months after operation were observed by imaging data. There was no significant difference in incision length, operation time, intraoperative bleeding volume, postoperative drainage volume and VAS score of lumbar incision 72 hours after operation among three groups (P>0.05). All cases were followed up for 12 to 36 (23.70±4.52) months. The height of intervertebral space in three groups recovered significantly (P<0.05), at the last follow-up, there were different degrees of loss, the loss degree of single common fusion cage group>banana type fusion cage group>double common fusion cage group. At the last follow-up, the Cobb angle in the coronaland sagittal planes of the three groups were significantly improved (P<0.05). During the follow-up, there were 42 cases of fusion cage subsidence, including 26 cases in the single common cage group, 5 cases in the double common cage group, 11 cases in the banana cage group, the difference was statistically significant (P<0.05). At 12 months after operation, the interbody fusion rate was 83% in the single common cage group, 95% in the double common cage group and 90% in the banana cage group, the interbody fusion rate in the two common cage group and the banana cage group was better than that in the single common cage group. No obvious degeneration was observed in the adjacent segments. At the last follow-up, the JOA scores of the three groups were statistically significant (P<0.05). The incidence of single common fusion cage group was 10%(4 / 42), that of double common fusion cage group was 9%(4 / 43), and that of banana fusion cage group was 10%(4 / 39). There was no significant difference among the three groups. Through the intermuscular approach, single pedicle screw and contralateral facet screw were used for fixation, and single common fusion cage, double common fusion cage or banana type fusion cage were used for interbody grafting to treat single segment lumbar disease. Although the application of different fusion cage could not increase the axial strength of fixed segment, the speed of fusion was accelerated by increasing the contact area, and the quality of the fusion cage reduces the settlement of the cage and the loss of the height of the intervertebral space. Therefore, two interbody fusion cages implanted in one side are of positive clinical significance for the fixation of unilateral pedicle screw combined with contralateral facet screw, without prolonging the operation time and increasing the complications.

Interspecific hybridization between cultivated morels Morchella importuna and Morchella sextelata by PEG-induced double inactivated protoplast fusion.

The commercial production of Morchella mushrooms calls for urgent breeding of excellent varieties or strains with appropriate tools, such as protoplast fusion. However, the protoplast fusion in morels has not been studied. In this paper, interspecific hybridization between cultivated morels of M. importuna and M. sextelata by PEG-induced protoplast fusion was conducted. Apart from functional complementation of double inactivated protoplasts, the fusants were characterized by cultural and cultivated characters and molecular markers of random amplified polymorphic DNA (RAPD). The results suggested that the hybrids and their parents showed significant difference in their inoculum recovery time, mycelial growth rate, yield of cultivation and total amino acid content of ascocarps. Moreover, positive barrage reactions were observed between parental strains as well as between each parent and a hybrid line. A dendrogram created on the basis of RAPD fingerprints exhibited three major clusters, in which morel hybrids showed intra-cluster variations, M. sextelata #6 formed an out group, while M. importuna #4 was phylogenetically closer to morel hybrids. All the results demonstrated that real fusants were obtained in our study. Protoplast fusion may provide an ideal alternative for new strain selection, and thus will promote the healthy development of morel industry.

Breast cancer targeted/ therapeutic with double and triple fusion Immunotoxins

Target-specific transport of therapeutic agents holds promise to increase the efficacy of cancer treatment by decreasing injury to normal tissues and post treatment problems. HER2 is a tumor cell surface marker that is expressed in 25–30 % of breast cancer patients. The significant role of HER2 in cancer development and its biological feature makes it a highly appealing goal for the therapeutic treatment of cancer targeted therapy using HER2 monoclonal antibody. This approach is currently used as a special treatment against breast cancer in some research. In the present study, HER2 monoclonal antibody (mAb), (Herceptin) fused to PE38 by recombinant DNA technology and a new recombinant IT was developed. The scFv(Herceptin)-PE-STXA and scFv(Herceptin)-PE fusions cloned in pET28a and recombinant protein expression was carried out and then the proteins were purified. MCF-7 and SKBR-3 cells were used as HER2-negative and HER2-positive breast cancer cells, respectively. The cytotoxicity of its evaluated using MTT assay. The cell ELISA was used to determine the binding ability of immunotoxins (ITs) to the cell receptor and internalization and apoptosis were also assessed. The results revealed that cell cytotoxicity occurred in SKBR-3 cells in a dose-dependent manner but not in MCF-7 cells. It is possible that this ITs can attach to HER2-positive breast cancer cells and then, internalize and eradicate cancer cells by apoptosis. Here, we concluded that the recombinant ITs have therapeutic potential against HER2-positive breast cancer.

High-Resolution Remote Sensing Image Retrieval Based on Classification-Similarity Networks and Double Fusion

In high-resolution remote sensing image retrieval (HRRSIR), convolutional neural networks (CNNs) have an absolute performance advantage over the traditional hand-crafted features. However, some CNN-based HRRSIR models are classification-oriented, they pay no attention to similarity, which is critical to image retrieval; whereas others concentrate on learning similarity, failing to take full advantage of information about class labels. To address these issues, we propose a novel model called classification-similarity network (CSN) , which aims for image classification and similarity prediction at the same time. In order to further improve performance, we build and train two CSNs, and two kinds of information from them, i.e., deep features and similarity scores, are consolidated to measure the final similarity between two images. Besides, the optimal fusion theorem in biometric authentication, which gives a theoretical scheme to make sure that fusion will definitely lead to a better performance, is used to conduct score fusion. Extensive experiments are carried out over publicly available datasets, demonstrating that CSNs are distinctly superior to usual CNNs and our proposed “two CSNs + feature fusion + score fusion” method outperforms the state-of-the-art models.

Intelligent Fusion of Deep Features for Improved Waste Classification

In this article, we address the problem of an image-based automatic classification of waste materials. Given the large number of waste categories and the importance of proper management of waste materials, the problem is known to be critical and of a particular interest. To achieve reliable waste classification capability, we propose a novel approach, that we name double fusion, which optimally combines multiple deep learning models using feature and score-level fusion methods. The double fusion scheme ensures an optimized contribution of the deep models by, firstly, combining their capabilities in an early and late fusion scheme followed by a score-level fusion of the classification results obtained with early and late fusion methods. In total, we employ and compare six different fusion methods including two feature-level fusion schemes, namely (i) Discriminant Correlation Analysis and (ii) simple concatenation of deep features, and four late fusion methods, namely (i) Particle Swarm Optimization, (ii) Genetic modeling of deep features (iii) Induced Ordered Weighted Averaging and (iv) a baseline method where all the deep models are treated equally. Moreover, we also evaluate the performance of the individual deep models, and compare our results against state-of-the-art methods demonstrating a significant improvement of 3.58% over state-of-the-art.

A combinational convolutional neural network of double subnets for food-ingredient recognition

Deep convolutional neural networks (DCNNs) have become the dominant machine learning for visual object recognition. They have been widely used in food image recognition and have achieved excellent performance. However, not only are the food-ingredient datasets not easy to obtain, but also the scale is not big enough to learn a deep learning model. For small-scale datasets, this paper proposes a novel DCNN architecture, which constructs an up-to-date combinational convolutional neural network of double subnets (CBDNet) for automatic classification of food ingredients using feature fusion. The feature fusion is a component which aggregates subnets for more abundant and precise deep feature extraction. In order to improve classification accuracy, some useful strategies are adopted, including batch normalisation (BN) operation and hyperparameters setting. Finally, experimental results show that the CBDNet integrating double subnets, feature fusion and BN operation extracts better image features and effectively improves the performance of food-ingredient recognition.

A combinational convolutional neural network of double subnets for food-ingredient recognition

Deep convolutional neural networks (DCNNs) have become the dominant machine learning for visual object recognition. They have been widely used in food image recognition and have achieved excellent performance. However, not only are the food-ingredient datasets not easy to obtain, but also the scale is not big enough to learn a deep learning model. For small-scale datasets, this paper proposes a novel DCNN architecture, which constructs an up-to-date combinational convolutional neural network of double subnets (CBDNet) for automatic classification of food ingredients using feature fusion. The feature fusion is a component which aggregates subnets for more abundant and precise deep feature extraction. In order to improve classification accuracy, some useful strategies are adopted, including batch normalisation (BN) operation and hyperparameters setting. Finally, experimental results show that the CBDNet integrating double subnets, feature fusion and BN operation extracts better image features and effectively improves the performance of food-ingredient recognition.

Motion Recognition Algorithm in VR Video Based on Dual Feature Fusion and Adaptive Promotion

VR video recognition in complex environment, a motion recognition algorithm based on two-feature fusion and adaptive enhancement is proposed to solve the problems of inaccurate target position, target drift and recognition error caused by the vulnerability to light change, target rotation and occlusion. First, based on the spatio-temporal context (STC) mechanism, image sequence features are extracted through spatio-temporal context relationship and visual system characteristics to reduce the influence of light changes and occlusion on behaviors. Secondly, reliable feature point tracks are obtained through image feature point tracking and background track cutting, and a rich set of action descriptors (AD) are calculated from which local motion information, shape and static appearance information of the track are retained. After that, the principal component analysis operator is introduced to define the double feature fusion rules, and the STC feature and AD feature are combined to form a more accurate and complete feature representation. Finally, adaptive boosting algorithm (ABA) is used to train the classification through the new features obtained and complete the decision judgment of behavior and action. The experimental results show that the proposed algorithm has higher recognition accuracy and robustness compared with the current commonly used motion recognition methods, and can better adapt to complex background and motion changes.

Recreation of t(7;12)(q36;p13) in Human Induced Pluripotent Stem Cells Using CRISPR/Cas9 Generates a Useful Model for Studying Acute Myeloid Leukemia

Introduction Acute myeloid leukemia (AML) is the result of aberrant hematopoietic processes, such as enhanced proliferation, blocked differentiation, and dysregulated apoptosis of hematopoietic stem and progenitor cells, and frequently these changes are initiated by chromosomal translocations in leukemia. Efficient methods for modelling leukemia and to recreate leukemia-associated genetic aberrations, such as chromosome translocations, are therefore crucial for investigating how leukemia is initiated. Today, most such models are murine and usually based on introduction of fusion gene transcripts of interest under the control of a constitutive active promoter using lenti- or retroviral transduction rather than the chromosomal translocation itself. The aim of the current project was to create a human cellular model of a chromosomal translocation that is typically found in AML in children under 24 months of age, the translocation t(7;12)(q36;p13). This translocation has been associated with poor prognosis, and leads to a gene fusion MNX1-ETV6 but also aberrant MNX1 gene expression. Its mechanism for leukemia initiation is so far unknown, mainly due to lack of a suitable experimental model. Material and methods CRISPR/Cas9 was used to reconstruct the genetics of the t(7;12)(q36;p13) rearrangement in human induced pluripotent stem cells (iPSC) while maintaining the genomic architecture and regulatory elements. Ribonucleoprotein (RNP) complex was delivered by lipofection (Nucleofection, Amaxa 4D system) into undifferentiated iPSC (ChiPSC 22, Cellartis). An ATTO550 tag on tracrRNA/RNP complex was used to sort out positive cells by flow cytometry and then seeded as single-cells in 96-well plates. Genomic DNA from the single-cell derived iPSC clones were screened by PCR for the presence of the translocation and positive clones were verified with a FISH probe specific for t(7;12)(q36;p13) (Double Fusion Break Apart probe, Metasystem). RT-qPCR was used to detect and quantify the expression of MNX1-ETV6 fusion and MNX1 transcripts. Differentiation potential was tested with the Trilineage Differentiation and Hematopoietic Kits (STEMdiff, STEMCELL Technologies). Results Using CRISPR/Cas9, we could successfully generate iPSC with the t(7;12)(q36;p13) translocation. The translocation was confirmed using conventional karyotyping and FISH and the mRNA expression of the fusion was confirmed with RT-qPCR. No additional chromosomal aberrations were seen. The t(7;12)(q36;p13) iPSC showed similar growth and differentiation properties as the parental iPSC. They showed propensity to differentiate into all three germ layers, confirming their pluripotent stem cell properties. The potential for differentiation into hematopoietic progenitor cells was shown by expression of CD34+, CD43+ and CD45+. In AML with t(7;12)(q36;p13), MNX1 mRNA expression is increased and this may play a role for leukemia development. In the t(7;12)(q36;p13) iPSC, RT-qPCR indeed showed increased expression of MNX1 expression compared with iPSC without the translocation. This increase of MNX1 was not seen in murine adult bone marrow or fetal liver cells transduced with retrovirus expressing the MNX1-ETV6 fusion. Further characterization of the t(7;12)(q36;p13) iPSC, e.g. whole exome and transcriptome sequencing and engraftment potential in immunocompromised mice (NSG-SGM), is ongoing. Conclusion In summary, we have using CRISPR/Cas9 successfully created a t(7;12)(q36;p13) iPSC line with potential to differentiate into hematopoietic progenitor cells and with gene expression pattern similar to what is seen in human AML samples with the t(7;12)(q36;p13). The introduction of the MNX1-ETV6 fusion in its correct genomic context could recapitulate local gene regulation, making it superior to models based on lenti- or retroviral introduction of fusion genes transcripts. In conclusion, this created cell line will be a valuable tool to study the mechanisms behind t(7;12)(q36;p13) AML, a severe form of AML associated with poor prognosis. Disclosures No relevant conflicts of interest to declare.

Clinical, Immunophenotypic and Genetic Characteristics of Aggressive (non-Burkitt) B-Cell Lymphoma in a Real Life Cohort

Introduction: Diffuse large B-cell lymphoma (DLBCL) is an aggressive and heterogeneous disease characterized by recurrent genetic alterations. Several adverse prognostic factors are well known as International Prognostic Index (IPI) parameters or MYC and BCL2 and/or BCL6 gene rearrangements (High-Grade B-cell Lymphoma double/triple hit, HGBL-DH/TH, according to the 2016 WHO classification). Other factors are controversial as previous history of indolent lymphoma, cell of origin (COO), co-expression of MYC and BCL2 proteins (double expressor (DE) status), MYC partner gene if rearranged. The aim of this study is to describe a « real life » cohort of patients treated in our institution for aggressive (non-Burkitt) B-cell lymphoma and to evaluate the prognostic impact of phenotypic and genetic alterations. Methods: We collected clinical and histological characteristics of patients uniformly treated between January 2009 and June 2017 with rituximab R-CHOP/CHOP-like chemotherapy. Treatment was reinforced with high dose methotrexate in case of central nervous system (CNS) localization or HGBL subtype. All tumor samples were analysed for MYC and BCL2 protein expression by immunohistochemistry. BCL2, BCL6 and MYC break were analysed by Interphase Fluorescence In Situ Hybridization (FISH) with breakapart probes. When MYC was rearranged MYC partner gene was analysed with double fusion probes (MYC/IgH, MYC/IgL, MYC/IgK). Positron Emission Tomography (PET scanner) was performed with maximal Standardized Uptake Value (SUV max) at baseline in all patients except 7. Results: 242 patients were studied. Baseline characteristics were: median age: 61 y (18-88), IPI 3-5: 49.6%, CNS localization: 6.6%. With a median follow-up of 4.4 years, 5y-overall survival (OS) and 5y-progression free survival (PFS) were 72% (66.0-78.5) and 65% (58.5-71.3), respectively. SUV max at baseline was 17,1 (minimum value 2,4 - maximal value 39,3).Histological diagnoses were DLBCL-not otherwise specified (NOS): 57%, transformed DLBCL: 26%, HGBL: 7.9%, primary mediastinal B-cell lymphoma (PMBL): 4.1% and others: 5%. Using Hans algorithm, 48.8% were classified as germinal center (GC), 47.9% as non-GC, 3.3% could not be evaluated and 38% were DE. MYC rearrangement was detected in 36 cases (14.9%) with an immunoglobulin partner gene (IgH, IgL or IgK) in 24 (77.4%) of 31 evaluable cases. BCL2 and BCL6 rearrangement were observed in 45 cases (18.6%) and 62 (25.6%) cases respectively. Fifteen cases were HGBL DH/TH including 7 DH MYC/BCL2, 5 DH MYC/BCL6 and 3 TH MYC/BCL2/BCL6. Four cases were HGBL-NOS. IPI score 3-5 (p&lt;0.0001) and HGBL (p=0.03) were significantly associated with inferior OS. IPI score 3-5 (p&lt;0.0001), DE status (p=0.03), MYC-R (p=0.05) and BCL2-R (p=0.0014) were significantly associated with inferior PFS. Previous indolent lymphoma, COO and MYC partner gene had no impact on OS or PFS. Conclusions: This monocentric study in a large cohort of aggressive B-cell lymphoma patients treated in real life conditions confirms previous reports on the negative prognostic impact of DE status, MYC and BCL2 translocations and HGBL subtype. These genetic and immuno-histological characteristics should have implications in the design and interpretation of future clinical trials. Disclosures Le Bras: Takeda: Research Funding; Pfizer: Other: Travel grant; Jansen: Other: Travel grant. Belhadj:Celgene: Other: personal fees from Celgene, personal fees from Amgen, personal fees from Takeda, personal fees from Janssen, outside the submitted work. Haioun:servier: Honoraria; amgen: Honoraria; janssen cilag: Consultancy; takeda: Consultancy; gilead: Consultancy; novartis: Honoraria; celgene: Honoraria; roche: Consultancy; celgene: Consultancy.

Early Outcomes of Pes Planus Reconstruction Using Either Hamstring Allograft or Synthetic Ligament

Category:Hindfoot, Midfoot/ForefootIntroduction/Purpose:Maintenance of the medial longitudinal arch is crucial to efficient kinematics in gait. It is supported by as complex interplay of osseous structures, ligaments, extrinsic tendons and plantar fascia.Two ligaments critical to stabilising the medial longitudinal arch are the Spring (Calcaneofibular) and Naviculocuneiform ligaments. The Spring Ligament serves as a sling within the “Acetabulum Pedis” to orientate the Talus relative to the Calcaneus and Navicular. The Naviculocuneiform ligament is an important plantar tension band and a separate entity to the Posterior Tibial Tendon (PTT) insertion. Attenuation of these ligaments leads to Pes Planus deformity and subsequent defunctioning of the PTT. This study evaluates the functional and radiological outcomes of Spring and Naviculocuneiform ligament reconstruction using Hamstring (Semitendinosis) Allograft and Synthetic ligament (Internal BraceTM, Arthrex).Methods:33 consecutive Pes Planus reconstructions were performed between 11/11/2013 and 06/03/2018. All patients were followed up prospectively with serial radiographs and functional scores including; MOXFQ, EQF5D and VAS. Minimum follow up was six months.At the midpoint during the study there was transition to using Synthetic Ligament instead of allograft. This was due to availability and ease of use.Surgical technique for both reconstructions included a proximal medial gastrocnemius recession and medialising calcaneal osteotomy. For allograft reconstructions, a pre-tensioned Semitendinosis allograft was fixed proximally to Talar neck using a tenodesis screw and passed plantar to PTT and through a bone tunnel in the medial cuneiform. Synthetic ligaments were inserted from the Sustentaculum Tali to the medial cuneiform in a hammock fashion. In both reconstructions tendinopathic PTTs were excised and a Flexor Digitorum Longus transfer used.Results:There were 17 synthetic ligament reconstructions (6 male, 11 female) and 16 allograft reconstructions (6 male, 10 female). Groups were matched pre-operatively for age, functional scores and radiological markers (T test P values >0.05).At six months significantly better improvements were observed in the synthetic ligament group compared to allograft group with regards to VAS, MOXFQ pain score, Meary’s line, 1st metatarsal Talus angle, Talonavicular uncoverage angle and Hindfoot alignment (T test P < 0.05).Statistical significance was maintained at 12 months with the synthetic ligament providing a significantly better reduction of Meary’s line 1st Metatarsal Talar angle, Talonavicular uncoverage and hindfoot alignment. 2 patients were revised to double fusions in the allograft group and 1 patient revised in the synthetic ligament group.Conclusion:Statistically significant improved functional scores and radiological appearance can be found up to 1 year following Synthetic ligament reconstruction of the Spring and Naviculocuneiform ligaments when compared to Hamstring allograft.

Anti-tumor effects of engineered mesenchymal stem cells in colon cancer model.

BackgroundCell-based gene therapy is considered as a promising strategy for the treatment of human malignancy. In many different types of cancer, mesenchymal stem cells (MSCs) are observed as valuable and potential anti-cancer agents. However, the exact mechanisms of MSCs involved in tumor microenvironment are not well understood.AimOur aims are to elucidate the MSCs-mediated tumor microenvironment.Materials and methodsIn this study, colon cancer model was established by injecting the HT29 cells into the subcutaneous of right axilla of nude mice. We applied the human placenta-derived MSCs (hP-MSCs) armed with a double fusion gene containing the herpes simplex virus truncated thymidine kinase and firefly luciferase for treatment of colon cancer on days 10, 15, and 20 after HT29 cells injection. Molecular imaging methods were used for real-time imaging tumor progression and tracking transplanted hP-MSCs by bioluminescence imaging. Furthermore, proliferation and apoptosis-related proteins levels in colon cancer tissues were examined by immunofluorescence and Western blotting.ResultsOur results demonstrated that the administration of engineered hP-MSCs significantly inhibited the tumors and this effect was enhanced by ganciclovir application. Further analysis demonstrated the anti-tumor effect of engineered hP-MSCs in vivo depended on inhibiting tumor proliferation and inducing tumor apoptosis.ConclusionCollectively, this work showed that engineered hP-MSCs could inhibit colon cancer progression and metastasis by inducing tumor cell death and suppressing proliferation.

Connection Status Research of the Resistance Spot Welding Joint Based on a Rectangular Terminal Electrode

Rectangular terminal electrode is adopted in this research to conduct a resistance spot welding (RSW) process on stainless steel plate. The connection status of RSW joints under different welding current, time, and pressure were studied, and revealed how the rectangular terminal electrode shape and its dimensions influence the RSW joint dimensions. The process analysis results showed that the RSW nuggets welded with rectangular terminal electrode are normally elliptical in shape, and the dimensions of the long axis direction and the short axis direction have a certain proportion. As the welding current increases, the nuggets dimensions in long direction increase first and then decrease, and the internal grain structure also varies. As the welding time increases, the nugget size in long direction increase first and then steady and rarely splash occurs. As the welding pressure increase, the nugget dimensions in long direction increase first and then decrease, and the splash easily occurs under large welding pressure. However, when the welding pressure is too small, the unique adjacent double fusion nugget formed. By adopting proper welding parameters, the nugget size and quality can be controlled. This provides an important basis for the application of rectangular terminal electrode in RSW welding process.