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Purpose: Abnormal glycosylation often occurs in tumor cells. T-synthase (core 1 beta 1,3- galactosyltransferase, C1GALT1, or T-synthase) is a key enzyme involved in O-glycosylation. Although T-synthase is known to be important in human tumors, the effects of T-synthase and T-antigen on human tumor responses remain poorly defined. Methods: In this study, a T-synthase-specific short hairpin RNA (shRNA) or T-synthase-specific eukaryotic expression vector(pcDNA3.1(+)) was transfected into murine Osteosarcoma LM8 cells to assess the effects of T-synthase on T cells and cytokines. Results: The up-regulation of T-synthase promoted the proliferation of osteosarcoma cells in vitro, but it promoted the proliferation of tumor initially up to 2-3 weeks but showed significant growth inhibitory effect after 3 weeks post-implantation in vivo. Osteosarcoma cells with high T-synthase expression in vitro promoted the proliferation and inhibited the apoptosis of CD8+ T cells. Further, T-synthase upregulation promoted CD8+ T-cell proliferation and the increased production of CD4+ T cell-derived IFN-γ cytokines to induce the increased tumor lethality of CTLs. Conclusion: Our data suggest that high T-synthase expression inhibits tumor growth by improving the body's anti-tumor immunity. Therefore, using this characteristic to prepare tumor cell vaccines with high immunogenicity provides a new idea for clinical immunotherapy of osteosarcoma.

Wearable inertial motion capture, a new type of motion capture technology, mainly estimates the human posture in 3-D space through multi-sensor data fusion. The available method for sensor fusion are usually aided by magnetometers to remove the drift error in yaw angle estimation, which in turn limits their application in the presence of complex magnetic field environment. In this paper, an extended Kalman filter data fusion method is proposed to fuse the 9-axis sensor data. Meanwhile, heuristic drift reduction (HDR) method is used to calibrate the accumulated error of heading angle. In addition, the position in 3-D space is estimated by foot-mounted Zero-Velocity-Update (ZUPT) technique. Combining 3-D attitude and position, a biomechanical model of the human body is established to track the motion of real human body. The extended Kalman filter (EKF) algorithm and position estimation methods are benchmarked against the golden standard, optical motion capture system, for various indoor experiments. In addition, various outdoor experiments are also conducted to verify the reliability of the proposed method. The results show that the proposed algorithm outperforms the available attitude estimation model in motion tracking and is feasible for 3-D human motion capture.

Cerebral palsy is one of the main factors leading to children’s disability. A large number of such children have hand motor dysfunction, such as limited range of motion, abnormal gestures, etc. Our goal is to design a prototype of wearable gesture training equipment for such children. For this purpose, this paper presents the development of a wireless smart glove to facilitate the accurate measurement of finger movement through the integration of multiple IMU sensors. Commercially available gaming gloves are not fitted with sufficient sensors for this particular application and most of them can only provide one degree of freedom for finger flexion and extension. In this paper, a data glove integrated with nine degree of freedom inertial sensors in conjunction with complex multi-sensor data fusion is developed. In our method, a single/dual state measurement update switching extended Kalman filter is proposed to estimate the spatial attitude of each finger segment. Through traversing the kinematic chain, a tree type hand dynamic model based on joint constraints is established to realize the real-time gesture tracking. Furthermore, a visual interface is developed to display the tracking effect of gestures, and the reliability of our algorithm is verified by optical contrast experiments, the repeatability of joint angle is also evaluated by kinematic analysis. In general, all the experimental results demonstrated that our proposed framework can accurately track the 3-D hand motion. This glove will help quantify joint stiffness and monitor patient progression during the rehabilitation training process.

We performed a double-blind, double-dummy controlled study to compare the efficacy between recombinant human thrombopoietin (rhTPO) and eltrombopag in rapidly increasing the platelet counts in Chinese patients with immune thrombocytopenia (ITP). A total of 96 patients diagnosed with ITP for ≥6 months who had baseline platelet counts of

For the issue of beautification and capacity expansion of the Quick Response(QR) code, we proposed an algorithm based on sequential module modulation. First, the modules for the padding codewords are modulated by the module-based binarized background image. Then, to increase the storage capacity, low-pass textured patterns are designed for both the black modules and the white modules. The modules of the plain QR code are modulated by the second-level message. Finally, these modulated modules are further modulated by the L-channel of the background image in Lab color space. The module elimination parameter in the second modulation is optimized to maximize an objective function that accounts for both the visual quality and the decoding error. Experimental results demonstrate that, the proposed algorithm is superior to the reference method in terms of visual quality and capacity.

Usual diagnostic methods of leptomeningeal metastases (LM) in CerebroSpinal fluid (CSF), lack both specificity and sensitivity. The Veridex CellSearch® technique quantifying circulating tumour cells (CTCs) in blood was adapted to detect Tumour Cells (CSFTCs) in CSF from cancer patients with LM. CSF samples from 60 patients with established or suspected breast cancer or lung cancer LM and/or melanoma were evaluated. 5 mL CSF samples were collected on CellSave® preservative and analyzed within 3 days after CSF sampling. Gold Standard cytological analysis on 1 to 10 mL CSF samples from patients with established LM allowed sometimes the detection but usually not the quantification of TCs. In established LM, EpCAM+/cytokeratin+ or CD146+/HMW-MAA+ nucleated (DAPI+) cells were observed and enumerated with precision from one to up to 10 000 cells/mL. Their morphology on digital images galleries could be discriminant between breast and lung cancer. This methodology, established on a limited volume of CSF compared to the Gold Standard and allowing delayed processing, is of great interest in the diagnosis and follow-up of cancer patients with LM. The reliability of the method also opens new fields of investigation for other biological fluids and to precise the stem cell potential of metastatic cells in CSF.

Considering the security of message exchange between service robot and cloud, we propose to authenticate the message integrity based on RSA algorithm and digital signature. In the process of message transmission, RSA algorithm is used to encrypt message for service robot and decrypt message for cloud. The digital signature algorithm is used to authenticate the source of the message. The results of experiment have proved that the proposed scheme can guarantee the security of message transmission.

In this paper, we combine beautified QR (Quick Response) code with data hiding algorithm. The QR code beautification algorithm based on error correction mechanism is used to generate the beautified QR code. In the process of generating the beautified QR code, the QR code modules are removed with the error correction capability allowed. The verified QR code is added to the beautified QR code to prevent the tampering of the QR code. Thus, we can enhance the security of the QR code when user scans it. The original image covered by the QR code and the verified QR code are embedded in the beautified QR code by using the LSB (Least Significant Bit) data hiding method. The experimental results show that the beautified QR code generated by the proposed algorithm is superior to the reference method in visual quality and security.

For the problem of beautification and self-recovery of visual QR (Quick Response) code, this paper proposes two schemes. The first scheme uses data hiding and beautified QR code for self-recovery. By decoding this visual QR code, we can not only access the relevant links embedded in it but also restore the original image. However, the QR code may be maliciously tampered with by the attacker during transmission (e.g., redirected to a phishing website). For tampering protection and recovery of the QR code, the QR code is bound with the background image. This increases the security level when user scans a QR code. Experimental results demonstrate that the proposed algorithm is superior to the reference method in terms of visual quality.

Dense granule disorder is one of the most common platelet abnormalities, resulting from dense granule deficiency or secretion defect. This study was aimed to evaluate the clinical usefulness of the flow cytometric combination of mepacrine uptake/release assay and CD63 expression detection in the management of patients with suspected dense granule disorder. Over a period of 5 years, patients with abnormal platelet aggregation and/or reduced adenosine triphosphate (ATP) secretion suggestive of dense granule disorder were consecutively enrolled. The flow cytometric assays were systematically performed to further investigate dense granule functionality. Among the 26 included patients, 18 cases showed impaired mepacrine uptake/release and reduced CD63 expression on activated platelets, consistent with δ-storage pool deficiency (SPD). Another seven patients showed decrease in mepacrine release and CD63 expression but mepacrine uptake was normal, indicating secretion defect rather than δ-SPD. Unfortunately, ATP secretion could not be measured in 7 out of the 26 patients due to insufficient sample and/or severe thrombocytopenia. This test combination provides a rapid and effective method to detect the heterogeneous abnormalities of platelet dense granule by distinguishing between storage and release defects. This combination is particularly advantageous for severely thrombocytopenic patients and pediatric patients in which only minimal sample is required.

Background Allogeneic hematopoietic stem cell transplantation (HSCT), the most widely used potentially curable cellular immunotherapeutic approach in the treatment of hematological malignancies, is limited by life-threatening complications: graft versus host disease (GVHD) and infections especially viral infections refractory to antiviral drugs. Adoptive transfer of virus-specific T cells is becoming an alternative treatment for infections following HSCT. We report here the results of a phase I/II multicenter study which includes a series of adenovirus-specific T cell (ADV-VST) infusion either from the HSCT donor or from a third party haploidentical donor for patients transplanted with umbilical cord blood (UCB). Methods Fourteen patients were eligible and 11 patients received infusions of ADV-VST generated by interferon (IFN)-γ-based immunomagnetic isolation from a leukapheresis from their original donor (42.9%) or a third party haploidentical donor (57.1%). One patient resolved ADV infection before infusion, and ADV-VST could not reach release or infusion criteria for two patients. Two patients received cellular immunotherapy alone without antiviral drugs as a pre-emptive treatment. Results One patient with adenovirus infection and ten with adenovirus disease were infused with ADV-VST (mean 5.83 ± 8.23 × 103 CD3+IFN-γ+ cells/kg) up to 9 months after transplantation. The 11 patients showed in vivo expansion of specific T cells up to 60 days post-infusion, associated with adenovirus load clearance in ten of the patients (91%). Neither de novo GVHD nor side effects were observed during the first month post-infusion, but GVHD reactivations occurred in three patients, irrespective of the type of leukapheresis donor. For two of these patients, GVHD reactivation was controlled by immunosuppressive treatment. Four patients died during follow-up, one due to refractory ADV disease. Conclusions Adoptive transfer of rapidly isolated ADV-VST is an effective therapeutic option for achieving in vivo expansion of specific T cells and clearance of viral load, even as a pre-emptive treatment. Our study highlights that third party haploidentical donors are of great interest for ADV-VST generation in the context of UCB transplantation. (N° Clinical trial.gov: NCT02851576, retrospectively registered). Electronic supplementary material The online version of this article (doi:10.1186/s13045-017-0469-0) contains supplementary material, which is available to authorized users.

Online supplement. (DOCX 21 kb)

International audience; Adoptive antiviral cellular immunotherapy by infusion of virus-specific T cells (VSTs) is becoming an alternative treatment for viral infection after hematopoietic stem cell transplantation. The T memory stem cell (TSCM) subset was recently described as exhibiting self-renewal and multipotency properties which are required for sustained efficacy in vivo. We wondered if such a crucial subset for immunotherapy was present in VSTs. We identified, by flow cytometry, TSCM in adenovirus (ADV)-specific interferon (IFN)-γ+ T cells before and after IFN-γ-based immunomagnetic selection, and analyzed the distribution of the main T-cell subsets in VSTs: naive T cells (TN), TSCM, T central memory cells (TCM), T effector memory cell (TEM), and effector T cells (TEFF). In this study all of the different T-cell subsets were observed in the blood sample from healthy donor ADV-VSTs, both before and after IFN-γ-based immunomagnetic selection. As the IFN-γ-based immunomagnetic selection system sorts mainly the most differentiated T-cell subsets, we observed that TEM was always the major T-cell subset of ADV-specific T cells after immunomagnetic isolation and especially after expansion in vitro. Comparing T-cell subpopulation profiles before and after in vitro expansion, we observed that in vitro cell culture with interleukin-2 resulted in a significant expansion of TN-like, TCM, TEM, and TEFF subsets in CD4IFN-γ T cells and of TCM and TEM subsets only in CD8IFN-γ T cells. We demonstrated the presence of all T-cell subsets in IFN-γ VSTs including the TSCM subpopulation, although this was weakly selected by the IFN-γ-based immunomagnetic selection system.

Interleukin-6 (IL-6) is an important growth factor for myeloma cells. IL-6 promotes the survival and proliferation of multiple myeloma (MM) cells through the phosphorylated proteins, including STAT3, MAPK, and Akt. Chemical components that suppress the signaling proteins' phosphorylation have a potential role for MM therapy. We recently identified that baicalein, a component of Scutellaria radix, suppressed proliferation and induced apoptosis of myeloma cells by blocking IkappaB-alpha degradation followed by down-regulating IL-6 and XIAP gene expression. In the present study of four myeloma cell lines, namely U266, NOP2, AMO1, and ILKM2, we demonstrated that baicalein not only inhibited IL-6-mediated phosphorylation of signaling proteins, such as Jak, STAT3, MAPK, and Akt, but also inhibited the expression of their target genes, such as bcl-xl. Finally, baicalein facilitated myeloma cell proliferation inhibited by dexamethasone. In contrast, baicalin, another major flavonoid derived from Scutellaria radix, had no significant effect on IL-6-mediated protein phosphorylation. Baicalein had no effect on Akt phosphorylation induced by the insulin-like growth factor-1 (IGF-1) in NOP2 cells. Compared with PD98059, an MAPK inhibitor, baicalein exhibited a stronger inhibitory effect on Erk(1/2) phosphorylation. Our results demonstrate that baicalein is a potent inhibitor of protein phosphorylation induced by IL-6, and thus may be a useful agent for the treatment of MM.

Melanoma is the most frequent solid tumor associated with leptomeningeal metastasis (LM). The usual diagnostic tools, that is, cytomorphological assessment of cerebro-spinal fluid (CSF) and gadolinium-enhanced MRI of the entire neuraxis both lack effectiveness. The CellSearch® Veridex technology for the detection of circulating tumor cells (CTC) in blood was designed for the follow-up and prognosis of breast, prostate, colorectal, and lung cancer, which express EpCAM markers. We have previously adapted this technology to detect malignant cells in the CSF of breast cancer LM. Our objective here was to check if this technology would also allow the detection and the enumeration of CTC in the CSF of melanoma patients presenting with LM although melanoma does not express EpCAM markers. On the occasion of the intrathecal treatment of LM in 2 melanoma patients, 5 mL of CSF and 7.5 mL of blood were collected on CellSave® Preservative Tubes and analyzed within 3 days after CSF sampling using a melanoma-dedicated kit. The CellSearch® Veridex technology was then adapted to direct enrichment, enumeration, and visualization of melanoma cells in the CSF. CD146+, HMW-MAA+, CD34−, and CD45− cells with typical morphology could be observed and enumerated sequentially with reproducible results, corresponding to CSF melanoma cells (CSFMC). In contrast to the current gold standard cytomorphological analysis, this new approach allowed a precise quantification of CSFMC in all samples concomitantly analyzed. This methodology, established on a limited volume of sample and allowing delayed processing, could prove of great interest in the diagnosis and follow-up of melanoma patients with LM.

Abstract 5019 Background: The Wnt/β-catenin signaling pathway has been implicated in a variety of tumor cell survival and proliferation, including myeloma cells. It can facilitate myeloma cell proliferation by upregulating the expression of many related genes, such as c-myc, cyclin D1, MMP and so on. We have previously demonstrated that Baicalein, a important component of Scutellaria radix from Huang-Lian-Jie-Du-Tang(HLJDT), can inhibit cell proliferation and induce cell death in myeloma cell through suppressing NF-κB activation (Ma Z, et al., Blood, 2005, 105(8): 3312–3318). In clinic, we found that HLJDT had a very good effective in multiple myeloma(MM) patients treatment (data not show). But, the precise molecular mechanism of HLJDT exerts its anti-tumor effects remains poorly understood. Here we further evaluated alternative mechanisms which is responsible for inhibition myeloma cell proliferation and migration by baicalein. Methods: Myeloma cell lines, RPMI8226 and U266 cells, were used for the analysis. MTT assay was used to assess cell viability in RPMI8226 and U266 cells which were exposed to baicalein with different concentrations and time period. Immunofluorescence assay and western blot analysis were used to assess β-catenin protein expression in MM cell lines. Transwell chamber migration assay was used to assess migration ability of myeloma cells treated with different concentrations of baicalein. RT-PCR analysis was used to assess β-catenin, c-myc, cyclin D1 and integrin β7 mRNA expression. Results: Baicalein can inhibit the MM cell lines proliferation in a dose- and time-dependent manner as measured by MTT assay. Immunofluorescence images and western bloting revealed that treatment of baicalein decreased the β-catenin protein expression level in myeloma cells. The mRNA expression level of β-catenin, c-myc and cyclin D1 was also decreased by baicalein treatment. In addition, we found that baicalein has an ability to inhibit the migration of MM cell llines, RPMI8226 and U266 cell, by decreasing the integrin β7 mRNA expression in a dose-dependent manner. Conclusion: Baicalein can inhibit myeloma cell proliferation is related to downregulation of β-catenin, c-myc and cyclin D1 genes expression, and the inhibition of myeloma ia associated with suppression of integrin β7 gene expression. Thus, HLJDT may have a potential of clinical applications in MM patient treatment. Disclosures: No relevant conflicts of interest to declare.

Purpose: To investigate the effect of N-glycan-defective mammary adenocarcinoma cells on the polarization of macrophages. Methods: N-glycan-defective breast cancer cells (MA782 cells) were prepared by swainsonine (SW) treatment and the cytotoxicity of SW to MA782 cells was evaluated using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. The N-glycan-defective MA782 cells were co-cultured with bone marrow-derived macrophages (BMDMs) for 48 h in vitro, and then the BMDMs and the co-cultured supernatant were analyzed for macrophage phenotypic using FQRT-PCR, FCM and ELISA. Results: SW-treated MA782 cells expressed defective N-glycan on the cell surface in a dose-dependent manner (*p < 0.05). MTT assays showed that neither the 1 μg/mL nor 5μg/mL SW treatments showed significant inhibition of MA782 cell growth in vitro. The expression of iNOS and agr-1 in the 5 μg/mL SW-treated group were 4.75-fold higher and 3.7-fold lower than that in the untreated group, respectively (*p < 0.05). Mean fluorescence intensity of CD16/32 expressed in the cells treated with 5 μg/mL SW was significantly higher in comparison with the untreated group (65 vs. 7, *p < 0.05), though the percentage of CD16/32-positive cells were not significantly different. Furthermore, the expression of CD206 and dectin-1 in the 5 μg/mL SW-treated group was significantly decreased (3.1±0.3% and 4.1±1.1%, respectively) in comparison with the untreated group (40±3% and 8.9±1.2%, respectively, both p < 0.05). In addition, the 5 μg/mL SW-treated group secreted more TNF-alpha (350 ±25 pg/mL) and less IL-10 (89±7.2 pg/mL) than the untreated group (80 ±3 pg/mL and 150 ±10 pg/mL, respectively, both p < 0.05). Conclusion: N-glycan-defective MA782 cells can induce the differentiation of BMDM into proinflammatory M1 macrophages in vitro.