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Background Epstein Barr virus (EBV) plays a causal role in some diseases, including infectious mononucleosis, lymphoproliferative diseases and nasopharyngeal carcinoma. Detection of EBV infection has been shown to be a useful tool for diagnosing EBV-related diseases. In the present study, we compared the performance of molecular tests, including fluorescence in situ hybridization (FISH) and EBV real-time PCR, to those of serological assays for the detection of EBV infection. Methods Thirty-eight patients with infectious mononucleosis (IM) were enrolled, of whom 31 were diagnosed with a mild type, and seven were diagnosed with IM with haemophagocytic lymphohistiocytosis and chronic active EBV infection. Twenty healthy controls were involved in the study. The atypical lymphocytes in peripheral blood were detected under a microscope and the percentage of positive cells was calculated. EBV DNA load in peripheral blood was detected using real-time PCR. The FISH assay was developed to detect the EBV genome from peripheral blood mononuclear cells (PBMC). Other diagnosis methods including the heterophil agglutination (HA) test and EBV-VCA-IgM test, to detect EBV were also compared. SPSS17.0 was used for statistical analysis. Results In all, 5–41% atypical lymphocytes were found among the PBMC in mild IM patients, whereas 8–51% atypical lymphocytes were found in IM patients with haemophagocytic lymphohistiocytosis and chronic active EBV infection patients. There was no significant difference in the ratios of atypical lymphoma between patients of the different types. We observed that 71.2% of mild IM patients and 85.7% of IM patients with haemophagocytic lymphohistiocytosis and chronic active EBV infection patients were positive for EBV-VCA-IgM. EBV-VCA-IgM was negative in all healthy control subjects. In addition, 67.1% of mild IM patients tested heterophile antibody positive, whereas 71.4% of IM patients with haemophagocytic lymphohistiocytosis and chronic active EBV infection tested positive. EBV DNA detected using real-time PCR was observed in 89.5% of these IM patients. The EBV genome was detected by the FISH assay in 97.4% of the IM patients. The EB viral loads detected by FISH and real-time PCR increased with the severity of IM. The EBV genome was detected in almost all the PBMC of IM with haemophagocytic lymphohistiocytosis and chronic active EBV infection patients. Conclusion Molecular tests, including FISH and EBV real-time PCR, are more sensitive than serological assays for the detection of EBV infection. The FISH assay detecting EBV copies in unfractionated whole blood is preferable and superior to plasma real-time PCR in its reflection of the absolute viral burden circulating in the patients.

As technology advances, IC designs are getting more sophisticated, thus it becomes more critical and challenging to fix printability issues in the design flow. Running lithography checks before tapeout is now mandatory for designers, which creates a need for more advanced and easy-to-use techniques for fixing hotspots found after lithographic simulation without creating a new design rule checking (DRC) violation or generating a new hotspot. This paper presents a new methodology for fixing hotspots on layouts while using the same engine currently used to detect the hotspots. The fix is achieved by applying minimum movement of edges causing the hotspot, with consideration of DRC constraints. The fix is internally simulated by the lithographic simulation engine to verify that the hotspot is eliminated and that no new hotspot is generated by the new edge locations. Hotspot fix checking is enhanced by adding DRC checks to the litho-friendly design (LFD) rule file to guarantee that any fix options that violate DRC checks are removed from the output hint file. This extra checking eliminates the need to re-run both DRC and LFD checks to ensure the change successfully fixed the hotspot, which saves time and simplifies the designer’s workflow. This methodology is demonstrated on industrial designs, where the fixing rate of single and dual layer hotspots is reported.

Glutelins derived from oats were purified and hydrolyzed sequentially using alcalase proteases for production of antioxidant peptides. The antioxidant activities of oat glutelin peptides were evaluated using reducing power methods including hydroxyl and 1,1-diphenyl-2-pycrylhydrazyl (DPPH) radical-scavenging assays. The results demonstrated that oats glutelin peptide D-1a exhibited the highest antioxidant activity compared to the other hydrolysates. D-1a was identified to be His–Tyr–Asn–Ala–Pro–Ala–Leu (784.38 Da) by electrospray ionization mass spectrometry (ESI-MS/MS). Se purificó glutelina derivada de avena e hidrolizada secuencialmente utilizando proteasas de tipo alcalasa para la producción de péptidos antioxidantes. Se evaluó la actividad antioxidante de los péptidos de glutelina de avena utilizando métodos de poder reductor, como por ejemplo ensayos de barrido de radicales hidroxilo y 1,1-difenil-2-picrilhidrazil (DPPH). Los resultados demostraron que los péptidos de glutelina D-1a de la avena exhibieron la mayor actividad antioxidante en comparación con otros hidrolizados. Se identificó D-1a como His-Tyr-Asn-Ala-Pro-Ala-Leu (784,38 Da) mediante espectrometría de masa-ionización por electrospray (ESI-MS/MS).

The present study conducted enzymatic hydrolysis of naked oats glutelin by Alcalase ( FG).Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) result showed that the molecular weight of naked oats glutelin ranged from 14 to 97 kDa. Ion exchange chromatography was used to isolate and purify naked oats glutelin hydrolysates. The results showed that hydrolysates were purified into 4 fractions by ion exchange chromatography. Fraction D showed higher scavenging effect against hydroxyl radical (IC50 1.40 mg mL-1), and 2,2-Dipheyl-1-picrylhydrazyl (DPPH) radical (IC50 1.26 mg mL-1) than the other 3 fractions. Moreover, fraction D exhibited higher scavenging effect against hydroxyl and DPPH radical than the hydrolysates. Free radical scavenging capacity of naked oats glutelin hydrolysates was improved by isolation and purification.

Adult T-cell leukemia (ATL) is an aggressive malignancy caused by human T-cell lymphotropic virus-1. There is no accepted curative therapy for ATL. We have reported that certain ATL patients have increased Notch-1 signaling along with constitutive activation of the nuclear factor-κB pathway. Physical and functional interaction between these two pathways provides the rationale to combine the γ-secretase inhibitor compound E with the proteasome inhibitor bortezomib. Moreover, romidepsin, a histone deacetylase inhibitor, has demonstrated major antitumor action in leukemia/lymphoma. In this study, we investigated the therapeutic efficacy of the single agents and the combination of these agents in a murine model of human ATL, the MT-1 model. Single and double agents inhibited tumor growth as monitored by tumor size (P

Oats and rice grains were subjected to high hydrostatic pressure (HHP) treatment at 300∼600 MPa at different times, and changes in the microorganism counts, inactivation of enzymes and selected physicochemical properties were investigated. HHP treatment (600 MPa, 5 min) resulted in a significant decrease of total bacterial count. With pressure of 300∼500 MPa for 0∼15 min, lipase activity (LA) was inhibited and the reaction by HHP fitted a first-order kinetics model. HHP treatment (600 MPa, 5 min) inactivated 97.5% LA and 92.0% catalase activity of oats. Additionally, the pasting rate of rice and oats treated with 600 MPa for 5 min increased by 75% and 68%, respectively. Meanwhile, the time of pasting degree up to 85% were decreased by 46%, and heat absorption capacities were decreased by 36% and 43%, respectively. Practical Applications As a convenient food source, instant rice has many advantages that make them popular to people, such as being easy to carry, affordable, healthy, having a long shelf life, etc. With the development of modern food processing, refined flour and milled rice became the new type of health destroyer, sometimes leading people to nutrient imbalance and malnutrition. Oat is introduced into the diet as an alternative crop because of its nutritive and health-promoting value. Therefore, we studied oats and rice as raw material compounds to novel instant rice. This study was based on high pressure treatment on oats and rice, for which we investigated changes on organisms, enzyme inactivation and physicochemical properties. We hope to provide a theoretical basis for rice and oats industrial development and to facilitate their production.

Aimed at the molding of polymer nanostructure parts, the interface model between long- and short-chain polycarbonates (PC) and nickel mold inserts was established by the molecular dynamics method. The molecular mechanism of the replication capability of polymer nanostructure part molding was discussed by analyzing the migration and diffusion of the molecular chain, concentration profile, filling morphology evolution, interface binding energy, and filling rate of conventional injection molding (CIM) and rapid heat cycle molding (RHCM). The results show that nanostructures are filled mainly during the packing stage. A short-chain PC system has a low glass transition temperature (Tg) and viscosity, good fluidity, and a high filling rate, so the replication capability of its nanostructures is good. A long-chain PC system has a fast cooling rate in CIM, its molecular chain motion is blocked, the filling rate is low, and the interface binding energy is small, and so its nanostructures have poor replication capability. However, the high temperature at the nanostructures can be maintained for a long time in RHCM, which promotes Brownian motion in the molecular chains. Under the action of packing pressure, molecular chains can overcome entanglement barriers and viscous resistance. Thus, the polymer concentration profile and filling rate increase with increasing packing pressure, which can produce more van der Waals energy. Furthermore, the evolution process of polymer filling morphology is realized by the Brownian motion of chain segments under packing pressure, that is, the diffusion motion of the molecular chain along the direction of a tube composed of other chains around it. With the increase of temperature or pressure, the migration and diffusion of the molecular chain can be promoted, thus, the replication capability of nanostructure parts for mold cavities can be enhanced.

In this paper, the foam-like composite scaffolds composed of hydroxyapatite (HA) and carbon nanotubes (CNTs) were prepared by a new method, where a polymer impregnating method was used for porous HA-based scaffold and a chemical vapor deposition (CVD) method was used for the growth of CNTs from the HA-based scaffold. The process produces the CNTs/HA scaffolds that have a foam-like structure with better mechanical property, better microstructure and a high degree of interconnection. A favorable pore size with big pores of 1–2 mm and small pores of 20–300 μm for osteoconduction and bone ingrowth is presented in these scaffolds. About 2 wt% multi-walled CNTs with the diameter of 60–100 nm are observed to be in situ grown from deficient nano-HA crystallites. Magnetic measurement exhibits these scaffolds are superparamagnetic with a saturation magnetization of 1.14 emu g −1 at a room temperature, benefiting the scaffolds to take up growth factors in vivo, stem cell or other bioactive molecules easily. This new type of CNTs/HA scaffolds is expected to have a promising applications in bone tissue engineering, targeted drug delivery system and other biomedical fields.

A health monitoring system is designed for smart management for personal health in this paper. Self-description and self-reasoning, the characteristics of ontology, make the link between machine and knowledge, knowledge point and the relevant knowledge point more closer. This system has certain application value on managing and analyzing basic health data. Database in this system is realized by storing user’s health data and disease information. Health monitoring system is user-centric which can record user’s health condition data and make recommendations for user according to the result of health data analyzing. User’s health status is expressed by a specific domain, like overweight and underweight. The experiment result shows that the health monitoring system can make a steady management for personal health on a long track record of health status and strict logic reasoning.

Most commonly used q -- ary stabilizer codes are derived from the codes over F q. We construct a nice error operator basis over the ring Z q, and consequently the quantum codes from self-orthogonal codes over Z q are described. We apply the building-up method for self-orthogonal codes over Z q, so that quantum (near)Maximum Distance Separable codes are obtained. By comparing the buiding-up method over the rings with the one over the fields, it is concluded that there are more choices of q with quantum codes over large finite ring Z q.

Discussed the "problem driven" teaching mode for the implementation of the basic steps and design issues of basic principles. Expounded the design of problem-based learning mode that based on different teaching objectives, teaching content, discussed the related teaching mode from five aspects, which are inspired problem driven, "bridge type" problem driven, multi angle problem driven, stepwise recursive problem driven and applied problem driven.

The aim of this study is to prepare Ca, P and Si-containing ternary oxide nagelschmidtite (NAGEL, Ca7Si2P2O16) bioceramics and explore their in vitro bioactivity for potential bone tissue regeneration. We prepared dense NAGEL ceramics through high-temperature sintering of NAGEL ceramic powders. The apatite-mineralization ability, dissolution rate, and human osteoblast response (including cytotoxicity analysis, attachment, morphology, proliferation, and bone-related gene expression) to NAGEL ceramics have been systematically studied by comparing with conventional β-tricalcium phosphate (β-TCP) ceramics. The results showed that NAGEL ceramics possessed more obvious apatite mineralization and dissolution (degradation) and stimulated bone-related gene expression (OCN and OPN) of osteoblasts than β-TCP ceramics. NAGEL ceramics also showed no significant cytotoxicity. NAGEL ceramics supported osteoblast attachment, proliferation, and osteogenic gene expression, with a comparable cell proliferation activity with β-TCP ceramics. These results indicate that novel NAGEL bioceramics with the specific composition of Ca7Si2P2O16, are a promising biomaterial for bone tissue regeneration application.

article i nfo In this work, the potential of Sr-hardystonite (Sr2ZnSi2O7) ceramics for biomedical use was first detected. First, pure Sr2ZnSi2O7 powders were successfully synthesized by sol-gel method, and then Sr2ZnSi2O7 ceramics were prepared by sintering the powder compacts. The mechanical test showed that the bending strength and Young's modulus of Sr2ZnSi2O7 ceramics could reach 82 MPa and 44 GPa, respectively, which were close to the values for human cortical bone. Degradation test in Tris-HCl buffer solution showed that Sr2ZnSi2O7 ceramics had a low degradation rate with less than 3% weight loss after soaking for 28 days. Furthermore, the in vitro biocompatibil- ity of the ceramics was evaluated by rabbit bone marrow stem cells (rBMSCs) adhesion and proliferation assay. The results showed that the ceramics supported the cells adhesion and proliferation. Taken together, Sr2ZnSi2O7 might be a potential candidate for preparation of bone implants.

Calcium silicate (CaSiO3) bioceramics and polyesters have complementary qualities as potential bone substituted materials. In this study, sintered CaSiO3 bioceramics were prepared and coated with poly(lactic-co-glycolic acid) (PLGA), and the influences of the PLGA coating on the degradation, hydrophilicity, bioactivity, and biocompatibility of CaSiO3 ceramics were investigated. The results showed that the degradation rate was reduced, while hydrophilicity was decreased with the increase of the polymer coating. In addition, the polymer coating resulted in a decrease of the alkaline pH value during the degradation of the ceramics, which indicated an increase of the cell biocompatibility, confirmed by the attachment and proliferation of rMSCs on the surface of the polymer-coated ceramics. Furthermore, the apatite-forming ability of the PLGA-coated CaSiO3 bioceramics was maintained. This study suggested that the coating with PLGA might be a useful method to improve the integrative properties of CaSiO3 bioceramics for applications in bone regeneration and bone tissue engineering.

The retrovirus, human T-cell–lymphotrophic virus-1 (HTLV-I) is the etiologic agent of adult T-cell leukemia (ATL) and the neurological disorder HTLV-I–associated myelopathy/tropical spastic paraparesis (HAM/TSP). The HTLV-I–encoded protein tax constitutively activates interleukin-2 (IL-2), IL-9, and IL-15 autocrine/paracrine systems that in turn activate the Jak3 (Janus kinase 3)/STAT5 (signal transducers and activators of transcription 5) pathway, suggesting a therapeutic strategy that involves targeting Jak3. We evaluated the action of the Jak3 inhibitor CP-690,550 on cytokine dependent ex vivo proliferation that is characteristic of peripheral blood mononuclear cells (PBMCs) from select patients with smoldering or chronic subtypes of ATL, or from those with HAM/TSP whose PBMCs are associated with autocrine/paracrine pathways that involve the production of IL-2, IL-9, IL-15, and their receptors. CP-690,550 at 50nM inhibited the 6-day ex vivo spontaneous proliferation of PBMCs from ATL and HAM/TSP patients by 67.1% and 86.4%, respectively. Furthermore, CP-690,550 inhibited STAT5 phosphorylation in isolated ATL T cells ex vivo. Finally, in an in vivo test of biological activity, CP-690,550 treatment of mice with a CD8 T-cell IL-15–transgenic leukemia that manifests an autocrine IL-15/IL-15Rα pathway prolonged the survival duration of these tumor-bearing mice. These studies support further evaluation of the Jak3 inhibitor CP-690,550 in the treatment of select patients with HTLV-I–associated ATL and HAM/TSP.

In 2003 Matveev suggested a new version of the Diamond Lemma suitable for topological applications. We apply this result to different situations and get a new conceptual proof of theorem on decomposition of three-dimensional manifolds into boundary connected sum of prime components.

In this study, the hydroxyapatite/wollastonite (Ca10(PO4)6(OH)2/CaSiO3, HAp/CS) composite bioceramics with different weight ratio were fabricated. The effects of composite ratio on sintering behavior, microstructure, mechanical properties, bioactivity, degradability behavior and the bone marrow mesenchymal stem cells (MSC) response to the composites were investigated. When the weight ratio of CS increased, the linear shrinkage of the ceramics decreased, while the porosity increased. The bending strength of the composites could be regulated between 98.06 ± 3.27 and 221.30 ± 15.69 MPa, and increased apparently with the increase of the CS component amount. The elastic modulus of the sintered samples was about 14.88–18.95 GPa, which was similar to that of human cortical bone. The bioactivity of the composites was enhanced with increasing CS content. For composites with more than 30 wt% CS contents, the samples were completely covered by a layer of dense apatite only after 1 day soaking. The dissolution rate of the samples increased with the increase of CS content, which suggested that the degradability of the HAp/CS composite bioceramics could be tailored by adjusting the initial HAp/CS ratio. In addition, the proliferation of MSC on the composites was examined and the results showed that higher content of CS content in composites promoted cell proliferation. When the CS content in the composite increased to 30 wt%, the proliferation rate of MSC cells showed significant higher than that of pure HAp (P

Hollow calcium silicate hydrate (CSH) microspheres with diameters around 1 microm were synthesized by a surfactant-assisted sonochemical route, and the products were characterized by XRD, SEM, FETEM and BET techniques. The results suggested that the ultrasound radiation, surfactant and Ca source were important factors which affected the formation of hollow microspheres. Based on the observation of products in different reaction systems, the possible mechanism for the formation of hollow CSH spheres was discussed. Furthermore, gentamicin, a typical anti-inflammatory drug, was used to investigate the drug loading and release behavior of the hollow spheres. The results indicated that CSH hollow spheres had high drug loading capacity and favorable drug release behavior, and might be used for preparation of bone grafts with drug delivery properties.

After neonatal HTLV-I infection through breast feeding, approximately 5% of HTLV-I carriers eventually develop Adult T-Cell Leukemia/Lymphoma (ATLL) with a latency of ~50 years, suggesting that acquired genetic and epigenetic changes in cellular genes act in concert with HTLV-I to initiate and maintain oncogenic transformation. We and others have recently utilized next generation sequencing technology to identify mutated genes that could be pivotal in the pathogenesis of ATLL. However, due to the complexity of genomic/epigenetic alteration in the ATLL genome, the identification of indispensable genes for proliferation and/or survival of ATLL cells remains a formidable challenge. To discover essential regulatory networks that are required for the proliferation and survival of ATLL cells, we performed a pooled shRNA screen in 8 ATLL cell lines using a library enriched for shRNAs targeting lymphoid regulatory factors and discovered that two BATF3 shRNAs and one IRF4 shRNA were highly toxic for all ATLL lines, but had little if any effect in other T cell and B cell lines. It is recently shown that a transcriptional complex of Irf4 and Batf binds to AP1-IRF composite (AICE) DNA motifs and plays key roles in the differentiation and function of certain mouse helper T cell subsets. A close paralogue of Batf, Batf3, is an indispensable transcription factor in a mouse dendritic cell subset, but also appears to play a redundant role with Batf in the differentiation of TH2 cells and can substitute for Batf in Batf knockout T cells. Our observations from shRNA screening suggested that IRF4 and BATF3 may cooperate to drive a transcriptional program that is essential for ATLL viability. We next used genome-wide chromatin precipitation (ChIP-seq) to identify the loci that are bound by BATF3 and IRF4. The set of binding peaks and the associated genes in IRF4 and BATF3 ChIP-seq intersected significantly. By integrating the ChIP-seq and gene expression profiling data of shBATF3- and shIRF4-ATLL cells, we defined a set of 68 BATF3-IRF4 direct target genes. Gene set enrichment analysis using gene expression profiling data from primary T cell lymphomas demonstrated that BATF3-IRF4 direct target genes were significantly enriched among genes that are more highly expressed in ATLL than in peripheral T-cell lymphoma, not otherwise specified (PTCL-NOS), suggesting that the BATF3 and IRF4 cooperatively regulate transcription in primary ATLL cells. HBZ is unique among HTLV-I viral proteins in being maintained in expression in all ATLL cases, suggesting that it may help maintain the malignant phenotype. Given that BATF3 and IRF4 are essential regulators in ATLL, we hypothesized possible relationship between HBZ and BATF3-IRF4 complex. We defined HBZ direct target genes by integrating the ChIP-seq and gene expression profiling data of HBZ-knockout ATLL cell lines by CRISPR/Cas9. Notably we discovered that BATF3 was among these. BATF3 mRNA and protein expression decreased following HBZ inactivation. The above considerations suggested that pharmacologic inhibition of the BATF3-IRF4 regulatory network might be a means to attack the HBZ oncogenic program therapeutically. ChIP-seq analysis of two enhancer marks, H3K27ac and BRD4, identified super-enhancers at the BATF3 locus in two ATLL cell lines. The small molecule JQ1 prevents the BET-protein BRD4 from interacting with chromatin, which is required for the function of super-enhancers. JQ1 treatment reduced BATF3 mRNA and protein levels in all ATLL lines tested, correlating with the eviction of BRD4 from the BATF3 super-enhancer. MYC mRNA and protein expression was also broadly downmodulated by JQ1. JQ1 treatment was consistently toxic for all ATLL cell lines tested at dose ranges that killed cell line models of T-ALL and DLBCL, which are known to rely on BET-proteins. In a dose-dependent manner, JQ1 also reduced the viability of primary ATLL samples and downregulated their expression BATF3 and MYC mRNA. Finally, we treated mouse xenograft models of ATLL with the BET-protein inhibitor CPI-203, a JQ1 analog with superior bioavailability in mice. In two different xenograft models, we observed significant tumor regression or growth inhibition, without evidence of systemic toxicity. Our study demonstrates that the HTLV-I virus exploits a regulatory module that can potentially be attacked therapeutically with BET protein inhibitors. Disclosures Yu: Celgene Corporation: Employment.

In this research, β-CaSiO3 particles were surface modified with dodecyl alcohol, and Poly-(DL-lactic acid) (PDLLA)/modified β-CaSiO3 composite films were fabricated with a homogenous dispersion of β-CaSiO3 particles in the PDLLA matrix. The aim of the study was to investigate the properties of the composite films before and after hydrolytic treatment. SEM images showed retained homogenous dispersion of β-CaSiO3 particles after hydrolysis and tensile test also showed maintained mechanical property. Simulated body fluid (SBF) incubation experiment suggested that hydrolytic treatment did not affect the formation of hydroxyapatite on the surface of the composite films. The hydrophilicity of the composites was greatly recovered (from 69.82° to 50.28°) after hydrolysis. In addition, cells cultured on composite films after hydrolysis presented the highest cell proliferation rate and differentiation level. All of these results suggested that the surface modification of silicate particles with dodecyl alcohol along with reversible hydrolytic treatment was an effective and feasible approach to fabricate polymer/silicate composite materials with improved properties.

The equilibrium solubility of the quaternary system RbCl-PrCl3-HCl-H2O was determined at 298.15 K and the corresponding equilibrium diagram was constructed in this paper. The quaternary system is complicated with three equilibrium solid phases, RbCl, RbPrCl4 · 4H2O (1:1 type) and PrCl3 · 6H2O, of which the new compound RbPrCl4 · 4H2O was found to be congruently soluble in the system. The new compound obtained was identified and characterized by the methods of X-ray diffraction, thermogravimetry, and differential thermogravimetry. The compound loses its crystal water by one step at 343 K to 453 K. The standard molar enthalpy of solution of RbPrCl4 · 4H2O in deionized water was measured to be −24.53 ± 0.22 kJ mol−1 by heat conduction microcalorimetry. Its standard molar enthalpy of formation was calculated to be −2743.20 ± 1.09 kJ mol−1.

Radioimmunotherapy of cancer with radiolabeled antibodies has shown promise. We evaluated an anti-CD25 monoclonal Antibody, 7G7/B6, armed with (90)Y as a potential radioimmunotherapeutic agent for CD25-expressing lymphomas.The lymphoma model was established by subcutaneous injection of 1 x 10(7) SUDHL-1 cells into nude mice. The biodistribution of (111)In-7G7/B6 and therapeutic studies with (90)Y-7G7/B6 were performed in the tumor-bearing mice.Therapy using (90)Y-7G7/B6 prolonged survival of the SUDHL-1 lymphoma-bearing mice significantly, as compared with either untreated mice or the mice treated with (90)Y-11F11, a radiolabeled isotype-matched control antibody (p0.001). All of the mice in the control and the (90)Y-11F11 treatment groups died by days 18 and 24, respectively. In contrast, 30% of the mice in the low-dose group (75 microCi of (90)Y-7G7/B6/mouse) and 75% in the high-dose group (150 microCi of (90)Y-7G7/B6/mouse) became tumor free and remained healthy for greater than 6 months.Our findings suggested that (90)Y-7G7/B6 is a potentially useful radioimmunotherapeutic agent for the treatment of patients with CD25-expressing lymphomas.

IL-15 has potential as an immunotherapeutic agent for cancer treatment because it is a critical factor for the proliferation and activation of natural killer (NK) and CD8 + T cells. Administration of anti-CD40 antibodies has shown anti-tumor effects in vivo through a variety of mechanisms. Furthermore, activation of CD40 led to increased expression of IL-15 receptor-α by dendritic cells, an action that is critical for trans-presentation of IL-15 to NK and CD8 + T cells. In this study, we investigated the therapeutic efficacy of the combination regimen of murine IL-15 (mIL-15) with an agonistic anti-CD40 antibody (FGK4.5) in murine lung metastasis models involving CT26 and MC38, which are murine colon cancer cell lines syngeneic to BALB/c and C57BL/6 mice, respectively. Treatment with mIL-15 or the anti-CD40 antibody alone significantly prolonged survival of both CT26 and MC38 tumor-bearing mice compared with the mice in the PBS solution control group ( P < 0.01). Furthermore, combination therapy with both mIL-15 and the anti-CD40 antibody provided greater therapeutic efficacy as demonstrated by prolonged survival of the mice compared with either mIL-15 or the anti-CD40 antibody-alone groups ( P < 0.001). We found that NK cells isolated from the mice that received the combination regimen expressed increased levels of intracellular granzyme B and showed stronger cytotoxic activity on the target cells. The findings from this study provide the scientific basis for clinical trials using the combination regimen of IL-15 with an anti-CD40 antibody for the treatment of patients with cancer.

Legume seeds contain 7S and/or 11S globulins as major storage proteins. The amino acid sequences of them from many legumes are similar to each other in the species but different from each other, meaning that some of these proteins from some crops exhibit excellent functional properties. To demonstrate this, we compared protein chemical and functional properties (thermal stability, surface hydrophobicity, solubility as a function of pH, and emulsifying properties) of these proteins from pea, fava bean, cowpea, and French bean with those of soybean as a control at the same conditions. The comparison clearly indicated that the 7S globulin of French bean exhibited excellent solubility (100%) at pH 4.2-7.0 even at a low ionic strength condition (mu = 0.08) and excellent emulsion stability (a little phase separation after 3 days) at pH 7.6 and mu = 0.08, although the emulsions from most of the other proteins separated in 1 h. These results indicate that our assumption is correct.

CD30 is a member of the TNF receptor superfamily. Overexpression of CD30 on some neoplasms versus limited expression on normal tissues makes this receptor a promising target for antibody-based therapy. Radioimmunotherapy of cancer with radiolabeled antibodies has shown promise. In this study, we evaluated the therapeutic efficacy of an anti-CD30 antibody, HeFi-1, armed with 211 At in a leukemia (karpas299) model and with 90 Y in a lymphoma (SUDHL-1) model. Furthermore, we investigated the combination therapy of 211 At-HeFi-1 with unmodified HeFi-1 in the leukemia model. Treatment with unmodified HeFi-1 significantly prolonged the survival of the karpas299-bearing mice compared with the controls ( P < 0.001). Treatment with 211 At-HeFi-1 showed greater therapeutic efficacy than that with unmodified HeFi-1 as shown by survival of the mice ( P < 0.001). Combining these two agents further improved the survival of the mice compared with the groups treated with either 211 At-HeFi-1 ( P < 0.05) or unmodified HeFi-1 ( P < 0.001) alone. In the lymphoma model, the survival of the SUDHL-1-bearing mice was significantly prolonged by the treatment with 90 Y-HeFi-1 compared with the controls ( P < 0.001). In summary, radiolabeled HeFi-1 is very promising for the treatment of CD30-expressing leukemias and lymphomas, and the combination regimen of 211 At-HeFi-1 with unmodified HeFi-1 enhanced the therapeutic efficacy.

Cancer dormancy is a stage in tumor progression in which residual disease remains occult and asymptomatic for a prolonged period. Cancer cell dormancy is the main cause of cancer recurrence and failure of therapy. However, cancer dormancy is poorly characterized and the mechanisms of how cancer cells develop dormancy and relapse remain elusive. In this study, 5- fluorouracil (5-FU) was used to induce cancer cell dormancy. We found that cancer cells escape the cytotoxicity of 5-FU by becoming "dormant". After exposure to 5-FU, residual non-small cell lung cancer (NSCLC) cells underwent epithelial-mesenchymal transition (EMT), followed by mesenchymal-epithelial transition (MET). These EMT-transformed NSCLC cells were in the state of cell quiescence where cells were not dividing and were arrested in the cell cycle in G0-G1. The dormant cells underwent an EMT showed characteristics of cancer stem cells. P53 is strongly accumulated in response to 5-FU-induced dormant cells through the activation of ubiquitin ligase anaphase-promoting complex (APC/C) and TGF-β/Smad signaling. In contrast to the EMT-transformed cells, MET-transformed cells showed an increased ability to proliferate, suggesting that dormant EMT cells were reactivated in the MET process. During the EMT-MET process, DNA repair including nonhomologous end joining (NHEJ) and homologous recombination (HR) is critical to dormant cell reactivation. Our findings provide a mechanism to unravel cancer cell dormancy and reactivation of the cancer cell population.

Haploid mammalian embryonic stem cells (ESCs) hold great promise for functional genetic studies and assisted reproduction. Recently, rodent androgenetic haploid ESCs (AG-haESCs) were generated from androgenetic blastocysts and functioned like sperm to produce viable offspring via the intracytoplasmic AG-haESCs injection into oocytes. However, the efficiency of this reproduction was very low. Most pups were growth-retarded and died shortly after birth, which is not practical for producing knockout animals. Further investigation suggested a possible link between the low birthrate and aberrant expression of imprinted genes. Here, we report the high-frequency generation of healthy, fertile mice from H19-Igf2 imprinting-locus modified AG-haESCs, which maintained normal paternal imprinting and pluripotency. Moreover, it is feasible to perform further genetic manipulations in these AG-haESCs. Our study provides a reliable and efficient tool to rapidly produce gene-modified mouse models and will benefit reproductive medicine in the future.

For a Hausdorff space X , we denote by 2 the collection of all closed subsets of X . In this paper, we discuss the connectedness and locally connectedness of hyperspace 2 endowed with the vietoris topology. Further path connectedness is investigated. The results generalize some theorems of E. Micheal. Keywords-connectedness; locally connectedness; path connectedness; vietoris topology; hyperspace

A series of Ni/SBA-15 catalysts with different Ni contents (2.5%–20%) were prepared using an incipient wetness impregnation method. The catalytic properties of the catalysts for methane reforming with carbon dioxide were tested using a continuous-flow fixed-bed quartz reactor. The structure of the catalysts was characterized using X-ray diffraction and N 2 adsorption. The results indicated that the Ni/SBA-15 catalysts led to a higher conversion of CH 4 and CO 2 under the reaction conditions. The 12.5% Ni/SBA-15 catalyst showed highly stable activity at 800°C for 600 h. However, after reaction for 710 h, the conversion of CH 4 and CO 2 decreased by about 50% and 25%, respectively. Coking was the main reason for the deactivation of the Ni/SBA-15 catalysts. At higher reaction temperatures, the mesoporous structure of SBA-15 was not destroyed and the pore walls of SBA-15 could prevent the aggregation of the nickel species. By inserting nickel into SBA-15, a small pore was formed besides the mesopores of SBA-15. The mesoporous structure of SBA-15 was not affected by the nickel species; however, the pore diameter, the pore volume, and the BET surface area decreased.

Radioimmunotherapy of cancer with radiolabeled antibodies has shown promise. α-Particles are very attractive for cancer therapy, especially for isolated malignant cells, as is observed in leukemia, because of their high linear energy transfer and short effective path length. We evaluated an anti-CD25 [interleukin-2 receptor α (IL-2Rα)] monoclonal antibody, 7G7/B6, armed with 211At as a potential radioimmunotherapeutic agent for CD25-expressing leukemias and lymphomas. Therapeutic studies were done in severe combined immunodeficient/nonobese diabetic mice bearing the karpas299 leukemia and in nude mice bearing the SUDHL-1 lymphoma. The results from a pharmacokinetic study showed that the clearance of 211At-7G7/B6 from the circulation was virtually identical to 125I-7G7/B6. The biodistributions of 211At-7G7/B6 and 125I-7G7/B6 were also similar with the exception of a higher stomach uptake of radioactivity with 211At-7G7/B6. Therapy using 15 μCi of 211At-7G7/B6 prolonged survival of the karpas299 leukemia–bearing mice significantly when compared with untreated mice and mice treated with 211At-11F11, a radiolabeled nonspecific control antibody (P < 0.01). All of the mice in the control and 211At-11F11 groups died by day 46 whereas >70% of the mice in the 211At-7G7/B6 group still survived at that time. In summary, 211At-7G7/B6 could serve as an effective therapeutic agent for patients with CD25-expressing leukemias. (Cancer Res 2006; 66(16): 8227-32)

A series of the Ce1−xCuxO2−x/Al2O3/FeCrAl catalysts (x = 0–1) were prepared. The structure of the catalysts was characterized using XRD, SEM and H2-TPR. The catalytic activity of the catalysts for the combustion of methane was evaluated. The results indicated that in the Ce1−xCuxO2−x/Al2O3/FeCrAl catalysts the surface phase structure were the Ce1−xCuxO2−x solid solution, α-Al2O3 and γ-Al2O3. The surface particle shape and size were different with the variety of the molar ratio of Ce to Cu in the Ce1−xCuxO2−x solid solution. The Cu component of the Ce1−xCuxO2−x/Al2O3/FeCrAl catalysts played an important role to the catalytic activity for the methane combustion. There were the stronger interaction among the Ce1−xCuxO2−x solid solution and the Al2O3 washcoats and the FeCrAl support.

Adult T-cell leukemia (ATL) develops in a small proportion of individuals infected with human T-cell lymphotrophic virus-1. The leukemia consists of an overabundance of activated T cells, which express CD25 on their cell surfaces. Presently, there is no accepted curative therapy for ATL. Flavopiridol, an inhibitor of cyclin-dependent kinases, has potent antiproliferative effects and antitumor activity. We investigated the therapeutic efficacy of flavopiridol alone and in combination with humanized anti-Tac antibody (HAT), which recognizes CD25, in a murine model of human ATL. The ATL model was established by intraperitoneal injection of MET-1 leukemic cells into nonobese diabetic/severe combined immunodeficient mice. Either flavopiridol, given 2.5 mg/kg body weight daily for 5 days, or HAT, given 100 μg weekly for 4 weeks, inhibited tumor growth as monitored by serum levels of human β-2-microglobulin (β2μ; P < .01), and prolonged survival of the leukemia-bearing mice (P < .05) as compared with the control group. Combination of the 2 agents dramatically enhanced the antitumor effect, as shown by both β2μ levels and survival of the mice, when compared with those in the flavopiridol or HAT alone group (P < .01). The significantly improved therapeutic efficacy by combining flavopiridol with HAT provides support for a clinical trial in the treatment of ATL.

Purpose: The use of an α emitter for radioimmunotherapy has potential advantages compared with β emitters. When administered systemically optimal targeting of intact antibodies requires >24 h, therefore limiting the use of short-lived α emitters. This study investigated the biodistribution of bismuth-labeled biotin in A431 tumor-bearing mice pretargeted with antibody B3-streptavidin (B3-SA) and examined the therapeutic efficacy of the α emitter, 213Bi-labeled biotin.Experimental Design: Biotinidase-resistant 7,10-tetraazacyclododecane-N,N′,N″,N‴-tetraacetic acid (DOTA)-biotin was radiolabeled with 205,206Bi or 213Bi. Treatment of tumor-bearing mice began by administration of B3-SA (400 μg) to target the tumor sites for 24 h. Then, an agent containing biotin and galactose groups was used to clear the conjugate from the circulation. Four h later, bismuth-radiolabeled DOTA-biotin was given, and biodistribution or therapy was evaluated. Dose escalation treatment from 3.7–74 MBq was performed, and the effects on tumors of different sizes were investigated. Tumor growth, complete blood cell counts, toxicity, and survival were monitored.Results: Radiolabeled biotin cleared rapidly. Rapid tumor uptake resulted in much higher tumor:nontumor targeting ratios than achieved with the directly labeled monoclonal antibody. Dose escalation revealed that 74 MBq caused acute death of mice, whereas 0.37–37 MBq doses inhibited tumor growth and prolonged survival significantly. Evidence of mild hematological toxicity was noted. At therapeutically effective doses renal toxicity was observed.Conclusions: 213Bi-DOTA-biotin, directed by the Pretarget method to tumor-targeted B3-SA, showed a therapeutic effect, although the therapeutic index was low. The source of the toxicity was most likely related to the renal toxicity.

Adult T-cell leukemia (ATL) develops in a small proportion of individuals infected with the retrovirus human T-cell leukemia virus (HTLV-1). We evaluated the efficacy of MEDI-507 (a humanized monoclonal antibody directed against CD2) alone and in combination with humanized anti-Tac (HAT) directed toward CD25, the interleukin-2 receptor α (IL-2Rα) using a human adult T-cell leukemia xenograft model. Weekly treatments (4) with HAT significantly prolonged the survival of the ATL-bearing mice when compared with phosphate-buffered saline (PBS)–treated controls (P < .0001). Mice treated with MEDI-507 (100 μg/wk for 4 weeks) survived longer than those treated with HAT (P < .0025). Furthermore, prolonged treatment (6 months) of ATL with MEDI-507 significantly improved the outcome when compared with a short course (4 weeks) of therapy (P < .0036). Such treatment with weekly MEDI-507 for 6 months led to a prolonged survival of the ATL-bearing mice that was comparable with the survival observed in the control group of mice that did not receive a tumor or therapeutic agent. We also found that the expression of Fcγ receptors (FcRγ) on polymorphonuclear leukocytes and monocytes was required for MEDI-507–mediated tumor killing in vivo. Thus, the tumor-killing mechanism with MEDI-507 in vivo required the expression of the receptor FcRγIII on polymorphonuclear leukocytes and monocytes, suggesting that it is mediated by a form of antibody-dependent cellular cytotoxicity. These results demonstrate that MEDI-507 has therapeutic efficacy on ATL in vivo and provides support for a clinical trial involving this monoclonal antibody in the treatment of patients with CD2-expressing leukemias and lymphomas. (Blood. 2003; 102:284-288)

Forward genetic screens using mammalian embryonic stem (ES) cells have identified genes required for numerous cellular processes. However, loss-of-function screens are more difficult to conduct in diploid cells because, in most cases, both alleles of a gene must be mutated to exhibit a phenotype. Recently, mammalian haploid ES cell lines were successfully established and applied to several recessive genetic screens. However, all these screens were performed in mixed pools of mutant cells and were mainly based on positive selection. In general, negative screening is not easy to apply to these mixed pools, although quantitative deep sequencing of mutagen insertions can help to identify some ‘missing’ mutants. Moreover, the interplay between different mutant cells in the mixed pools would interfere with the readout of the screens. Here, we developed a method for rapidly generating arrayed haploid mutant libraries in which the proportion of homozygous mutant clones can reach 85%. After screening thousands of individual mutant clones, we identified a number of novel factors required for the onset of differentiation in ES cells. A negative screen was also conducted to discover mutations conferring cells with increased sensitivity to DNA double-strand breaks induced by the drug doxorubicin. Both of these screens illustrate the value of this system.

A nitrate-dominant synthetic wastewater simulating slightly polluted water with low C/N and poor biochemical availability was treated in lab-scale vertical-flow (VF)-constructed wetlands, which had Phragmites australis planted with different types of external carbon sources: Platanus acerifolia leaf litters, P. australis litters, glucose and a blank test with no external carbon sources. A comparison of the TN removal and N2O flux performances among the four wetland reactors indicated higher TN removal efficiencies and N2O release fluxes in the VF wetland columns with external carbon sources, as measured by the percentage removal of TN (P. acerifolia leaf litters 82.49 %, P. australis litters 70.55 %, glucose 62.50 % and blank 46.45 %) and N2O flux (P. acerifolia leaf litters 2275.22 μg · m−2 · h−1, P. australis litters 1920.53 μg · m−2 · h−1, glucose 1598.57 μg · m−2 · h−1 and blank 1192.08 μg · m−2 · h−1). This was primarily because of an improved supply of organic carbon from the external carbon sources for heterotrophic denitrification. And, the nitrogen released from the decomposition of plant materials resulted in the N2O release fluxes to some extent. However, employing P. acerifolia leaf litters and P. australis litters as external carbon sources caused net increases in organics of the final effluent water. Overall, the results not only demonstrated the potential of using external plant carbon sources in VF wetlands to enhance the TN removal efficiency but also showed a risk of excessive organic release and greater N2O flux feedback to global warming. Hence, future studies are needed to optimise the quantity and method for adding external carbon sources to VF-constructed wetlands so that sufficient nitrate removal efficiency is achieved and the N2O flux and organic pollution are minimised.

Adhesion molecules play a major role in the processes of invasion and metastasis of malignant tumors. Their expression within tumors has been reported to be quantitatively and qualitatively altered according to the invasiveness and metastatic potential of the tumor. The present study tested whether the intratumoral expression of integrin alpha 3 can be detected by a radiolabeled monoclonal antibody. The in vitro binding study with four different human cancer cells showed that radioiodinated GA17 antibody recognizing integrin alpha 3 bound specifically to these cells to varying degrees, according to the antigen density on each cell. The biodistribution study with 125I- and 111In-labeled antibodies showed specific localization of radiolabeled GA17 to the xenografts. However, the in vivo tumor localization was not proportional to the antigen density calculated in vitro, and antibody metabolism varied among the tumors, as was also confirmed by in vitro radionuclide retention assay. The intratumoral distribution of radioactivities varied reflecting the antigen expression within the tumor. These results indicate that 1) integrin alpha 3 was expressed in various kinds of tumors and could be localized by the radiolabeled antibody, and 2) the expression of integrin alpha 3 and the metabolism of the radiolabeled antibody after binding to the antigen within the tumor were variable among the tumors, which affected the radionuclide distribution characteristics. The expression of adhesion molecules within these tumors was noninvasively detected by a radiolabeled antibody. It may be possible to use integrin alpha 3, when it is overexpressed, as a target of therapy with antibodies radiolabeled with alpha or beta emitters.

Although development of human anti-murine immunoglobulin antibody (HAMA) is often seen in patients receiving murine antibodies, the variety of methods used for detecting HAMA makes it difficult to compare directly the HAMA responses measured by different assays. In the present study, several parameters of the HAMA response to two murine monoclonal antibodies were evaluated. The anti-sialosyl Tn antibody MLS102 and anti-CA125 antibody 145-9, which were labeled with 111In, were injected intravenously into 17 colorectal cancer patients and 11 ovarian cancer patients for immunoscintigraphy, respectively. HAMA was measured by enzyme-linked immunosorbent assay. There was no difference in baseline HAMA levels before antibody injection between the two groups. HAMA developed more frequently in ovarian cancer patients receiving the 145-9 antibody than in colorectal cancer patients receiving the MLS102 antibody (9/11 vs. 6/17, P < 0.05). No significant difference was observed in maximal HAMA levels between the two groups of patients. However, time to reach the maximal levels was delayed and the duration of the response seemed longer in ovarian cancer patients. Among 11 patients receiving the 145-9 antibody three patients became positive for HAMA more than 2 months after antibody injection and the other two had HAMA activity in their sera for more than 17 months. HAMA response was different between the two antibodies, and late onset or long duration of HAMA response against the 145-9 antibody suggests the importance of HAMA measurement in patients who receive a second injection of murine antibodies even after a long interval.

Burkitt's lymphoma (BL) can often be cured by intensive chemotherapy, but the toxicity of such therapy precludes its use in the elderly and in patients with endemic BL in developing countries, necessitating new strategies(1). The normal germinal centre B cell is the presumed cell of origin for both BL and diffuse large B-cell lymphoma (DLBCL), yet gene expression analysis suggests that these malignancies may use different oncogenic pathways(2). BL is subdivided into a sporadic subtype that is diagnosed in developed countries, the Epstein-Barr-virus-associated endemic subtype, and an HIV-associated subtype, but it is unclear whether these subtypes use similar or divergent oncogenic mechanisms. Here we used high-throughput RNA sequencing and RNA interference screening to discover essential regulatory pathways in BL that cooperate with MYC, the defining oncogene of this cancer. In 70% of sporadic BL cases, mutations affecting the transcription factor TCF3 (E2A) or its negative regulator ID3 fostered TCF3 dependency. TCF3 activated the pro-survival phosphatidylinositol-3-OH kinase pathway in BL, in part by augmenting tonic B-cell receptor signalling. In 38% of sporadic BL cases, oncogenic CCND3 mutations produced highly stable cyclin D3 isoforms that drive cell cycle progression. These findings suggest opportunities to improve therapy for patients with BL.

Knowledge of oncogenic mutations can inspire therapeutic strategies that are synthetically lethal, affecting cancer cells while sparing normal cells. Lenalidomide is an active agent in the activated B-cell-like (ABC) subtype of diffuse large B cell lymphoma (DLBCL), but its mechanism of action is unknown. Lenalidomide kills ABC DLBCL cells by augmenting interferon β (IFNβ) production, owing to the oncogenic MYD88 mutations in these lymphomas. In a cereblon-dependent fashion, lenalidomide downregulates IRF4 and SPIB, transcription factors that together prevent IFNβ production by repressing IRF7 and also amplify pro-survival NF-κB signaling by transactivating CARD11. Blockade of B cell receptor (BCR) signaling using the BTK inhibitor ibrutinib also downregulates IRF4 and consequently synergizes with lenalidomide in killing ABC DLBCLs, suggesting attractive therapeutic strategies.

To investigate the relationship between the promoter polymorphism of IL6 (-174GC, -572GC and -597GA) and chronic rhinosinusitis (CRS).The case-control study consisted of 123 patients with CRS and 239 controls from a Chinese Han population from Shanghai. The genotypes of the subjects were determined by polymerase chain reaction-restriction fragment length polymorphism and gene sequencing. Besides, the concentrations of the totle immunoglobulin E (TIgE) and eosinophilic cationic protein (ECP) in the blood were also determined.The -174GC and -597GA polymorphisms were not detected in this study population. Significant differences in genotype and allele frequencies of -572C/G were observed between CRS patients and control groups. In CRS patients, the CC, CG, GG genotype frequencies were 69.1%, 29.3%, 1.6%, C, G allele frequencies were 83.7%, 16.3%. In control group, the genotype frequencies were 55.2%, 42.3%, 2.5%, the allele frequencies were 76.4%, 23.6%, respectively. The -572CC genotype was associated with an increased risk of developing CRS (P0.05, OR = 1.932, 95% CI, 1.205-3.097). There was no significant differences in the concentrations of the TIgE and ECP among each genotype.IL-6 gene -572GC polymorphism is associated with the susceptibility to CRS. CC genotype could be an independent risk factor.

IL-15 has potential as an immunotherapeutic agent for cancer treatment because of its ability to effectively stimulate CD8 T cell, natural killer T cell, and natural killer cell immunity. However, its effectiveness may be limited by negative immunological checkpoints that attenuate immune responses. Recently a clinical trial of IL-15 in cancer immunotherapy was initiated. Finding strategies to conquer negative regulators and enhance efficacy of IL-15 is critical and meaningful for such clinical trials. In a preclinical study, we evaluated IL-15 combined with antibodies to block negative immune regulator cytotoxic T lymphocyte antigen 4 (CTLA-4) and programmed death ligand 1 (PD-L1) in an established murine transgenic adenocarcinoma of mouse prostate (TRAMP)-C2 prostate tumor model. IL-15 treatment resulted in a significant prolongation of survival in tumor-bearing animals. Coadministration of anti-PD-L1 or anti-CTLA-4 singly with IL-15 did not improve animal survival over that of IL-15 alone. However, simultaneous administration of IL-15 with anti-CTLA-4 and anti-PD-L1 was associated with increased numbers of tumor antigen-specific tetramer-positive CD8 T cells, increased CD8 T-cell tumor lytic activity, augmented antigen-specific IFN-γ release, decreased rates of tumor growth, and improved animal survival compared with IL-15 alone. Furthermore, triple combination therapy was associated with inhibition of suppressive functions of CD4 + CD25 + regulatory T cells and CD8 + CD122 + regulatory T cells. Thus, simultaneous blockade of CTLA-4 and PD-L1 protected CD4 and/or CD8 T-cell activity from these regulatory T cells. Combining the immune stimulatory properties of IL-15 with simultaneous removal of two critical immune inhibitory checkpoints, we showed enhancement of immune responses, leading to increased antitumor activity.