Immunodepletion of MDSC by AMV564, a novel bivalent, bispecific CD33/CD3 T cell engager, ex vivo in MDS and melanoma.

We have reported previously that CD33hi myeloid-derived suppressor cells (MDSCs) play a direct role in the pathogenesis of myelodysplastic syndromes (MDSs) and that their sustained activation contributes to hematopoietic and immune impairment, including modulation of PD1/PDL1. MDSCs can also limit the clinical activity of immune checkpoint inhibition in solid malignancies. We hypothesized that depletion of MDSCs may ameliorate resistance to checkpoint inhibitors and, hence, targeted them with AMV564 combined with anti-PD1 in MDS bone marrow (BM) mononuclear cells (MNCs) enhanced activation of cytotoxic T cells. AMV564 was active in vivo in a leukemia xenograft model when co-administered with healthy donor peripheral blood MNCs (PBMCs). Our findings provide a strong rationale for clinical investigation of AMV564 as a single agent or in combination with an anti-PD1 antibody and in particular for treatment of cancers resistant to checkpoint inhibitors.

Oxidized Mitochondrial DNA Engages TLR9 to Activate the NLRP3 Inflammasome in Myelodysplastic Syndromes.

Myelodysplastic Syndromes (MDSs) are bone marrow (BM) failure malignancies characterized by constitutive innate immune activation, including NLRP3 inflammasome driven pyroptotic cell death. We recently reported that the danger-associated molecular pattern (DAMP) oxidized mitochondrial DNA (ox-mtDNA) is diagnostically increased in MDS plasma although the functional consequences remain poorly defined. We hypothesized that ox-mtDNA is released into the cytosol, upon NLRP3 inflammasome pyroptotic lysis, where it propagates and further enhances the inflammatory cell death feed-forward loop onto healthy tissues. This activation can be mediated via ox-mtDNA engagement of Toll-like receptor 9 (TLR9), an endosomal DNA sensing pattern recognition receptor known to prime and activate the inflammasome propagating the IFN-induced inflammatory response in neighboring healthy hematopoietic stem and progenitor cells (HSPCs), which presents a potentially targetable axis for the reduction in inflammasome activation in MDS. We found that extracellular ox-mtDNA activates the TLR9-MyD88-inflammasome pathway, demonstrated by increased lysosome formation, IRF7 translocation, and interferon-stimulated gene (ISG) production. Extracellular ox-mtDNA also induces TLR9 redistribution in MDS HSPCs to the cell surface. The effects on NLRP3 inflammasome activation were validated by blocking TLR9 activation via chemical inhibition and CRISPR knockout, demonstrating that TLR9 was necessary for ox-mtDNA-mediated inflammasome activation. Conversely, lentiviral overexpression of TLR9 sensitized cells to ox-mtDNA. Lastly, inhibiting TLR9 restored hematopoietic colony formation in MDS BM. We conclude that MDS HSPCs are primed for inflammasome activation via ox-mtDNA released by pyroptotic cells. Blocking the TLR9/ox-mtDNA axis may prove to be a novel therapeutic strategy for MDS.

Role of IL-9, IL-2RA, and IL-2RB genetic polymorphisms in coronary heart disease.

BACKGROUND: Coronary heart disease (CHD) is one of the leading causes of disability and death worldwide. Inflammatory cytokines play an essential role in the pathogenesis of CHD. This study aimed to detect the potential association between interleukin (IL)-9, IL-2RA, and IL-2RB variants and CHD in a Han Chinese population. METHODS: This case-control study included 499 CHD patients and 496 healthy controls. Seven single-nucleotide polymorphisms (SNPs) were genotyped to investigate the possible association between the polymorphisms and CHD risk. Interactions between SNPs and CHD risk were analyzed via multifactor dimensionality reduction (MDR). RESULTS: We observed an association between IL­9 rs55692658 (OR = 1.72, p = 0.003) and increased CHD risk. Age-stratified analysis indicated that regardless of the participants' age, IL­9 rs55692658 and IL-2RB rs1573673 contributed significantly to CHD susceptibility (p < 0.05, respectively). Results showed an association between IL­9 rs55692658 and an increased risk for CHD (OR = 2.32, p = 0.003), while IL-2RA rs12722498 was correlated with decreased susceptibility to CHD (OR = 0.54, p = 0.033) in female patients. Furthermore, IL-2RA rs12569923 was related to diabetes risk in CHD patients (OR = 1.50, p = 0.028). The MDR analysis revealed a positive interaction between the SNPs. CONCLUSION: The present study demonstrated that IL­9 rs55692658, IL-2RA rs12569923, IL-2RA rs12722498, and IL-2RB rs3218264 polymorphisms might be related to CHD. The results require validation in larger studies.

Constitutively Activated DAP12 Induces Functional Anti-Tumor Activation and Maturation of Human Monocyte-Derived DC.

Dendritic cells (DCs) are professional antigen presenting cells with a great capacity for cross-presentation of exogenous antigens from which robust anti-tumor immune responses ensue. However, this function is not always available and requires DCs to first be primed to induce their maturation. In particular, in the field of DC vaccine design, currently available methodologies have been limited in eliciting a sustained anti-tumor immune response. Mechanistically, part of the maturation response is influenced by the presence of stimulatory receptors relying on ITAM-containing activating adaptor molecules like DAP12, that modulates their function. We hypothesize that activating DAP12 in DC could force their maturation and enhance their potential anti-tumor activity for therapeutic intervention. For this purpose, we developed constitutively active DAP12 mutants that can promote activation of monocyte-derived DC. Here we demonstrate its ability to induce the maturation and activation of monocyte-derived DCs which enhances migration, and T cell stimulation in vitro using primary human cells. Moreover, constitutively active DAP12 stimulates a strong immune response in a murine melanoma model leading to a reduction of tumor burden. This provides proof-of-concept for investigating the pre-activation of antigen presenting cells to enhance the effectiveness of anti-tumor immunotherapies.

[Risk factors for metabolic bone disease of prematurity in very/extremely low birth weight infants: a multicenter investigation in China].

OBJECTIVE: To investigate the incidence rate and risk factors for metabolic bone disease of prematurity (MBDP) in very low birth weight/extremely low birth weight (VLBW/ELBW) infants. METHODS: The medical data of 61 786 neonates from multiple centers of China between September 1, 2013 and August 31, 2016 were retrospectively investigated, including 504 VLBW/ELBW preterm infants who met the inclusion criteria. Among the 504 infants, 108 infants diagnosed with MBDP were enrolled as the MBDP group and the remaining 396 infants were enrolled as the non-MBDP group. The two groups were compared in terms of general information of mothers and preterm infants, major diseases during hospitalization, nutritional support strategies, and other treatment conditions. The multivariate logistic regression analysis was used to investigate the risk factors for MBDP. RESULTS: The incidence rate of MBDP was 19.4% (88/452) in VLBW preterm infants and 38.5% (20/52) in ELBW preterm infants. The incidence rate of MBDP was 21.7% in preterm infants with a gestational age of < 32 weeks and 45.5% in those with a gestational age of < 28 weeks. The univariate analysis showed that compared with the non-MBDP group, the MBDP group had significantly lower gestational age and birth weight, a significantly longer length of hospital stay, and a significantly higher incidence rate of extrauterine growth retardation (P < 0.05). Compared with the non-MBDP group, the MBDP group had significantly higher incidence rates of neonatal sepsis, anemia, hypocalcemia, and retinopathy of prematurity (P < 0.05). The MBDP group had a significantly lower mean feeding speed, a significantly higher age when reaching total enteral feeding, and a significantly longer duration of parenteral nutrition (P < 0.05). The use rate of caffeine citrate in the MBDP group was significantly higher, but the use rate of erythropoietin was significantly lower than that in the non-MBDP group (P < 0.05). The multivariate logistic regression analysis showed that gestational age < 32 weeks, hypocalcemia, extrauterine growth retardation at discharge, and neonatal sepsis were risk factors for MBDP (P < 0.05). CONCLUSIONS: A lower gestational age, hypocalcemia, extrauterine growth retardation at discharge, and neonatal sepsis may be associated an increased risk of MBDP in VLBW/ELBW preterm infants. It is necessary to strengthen perinatal healthcare, avoid premature delivery, improve the awareness of the prevention and treatment of MBDP among neonatal pediatricians, and adopt positive and reasonable nutrition strategies and comprehensive management measures for preterm infants.

[A multicenter survey of antibiotic use in very and extremely low birth weight infants in Hunan Province].

OBJECTIVE: To investigate the current status of antibiotic use for very and extremely low birth weight (VLBW/ELBW) infants in neonatal intensive care units (NICUs) of Hunan Province. METHODS: The use of antibiotics was investigated in multiple level 3 NICUs of Hunan Province for VLBW and ELBW infants born between January, 2017 and December, 2017. RESULTS: The clinical data of 1 442 VLBW/ELBW infants were collected from 24 NICUs in 2017. The median antibiotic use duration was 17 days (range: 0-86 days), accounting for 53.0% of the total length of hospital stay. The highest duration of antibiotic use was up to 91.4% of the total length of hospital stay, with the lowest at 14.6%. In 16 out of 24 NICUs, the antibiotic use duration was accounted for more than 50.0% of the hospitalization days. There were 113 cases with positive bacterial culture grown in blood or cerebrospinal fluid, making the positive rate of overall bacterial culture as 7.84%. The positive rate of bacterial culture in different NICUs was significantly different from 0% to 14.9%. The common isolated bacterial pathogens Klebsiella pneumoniae was 29 cases (25.7%); Escherichia coli 12 cases (10.6%); Staphylococcus aureus 3 cases (2.7%). The most commonly used antibiotics were third-generation of cephalosporins, accounting for 41.00% of the total antibiotics, followed by penicillins, accounting for 32.10%, and followed by carbapenems, accounting for 13.15%. The proportion of antibiotic use time was negatively correlated with birth weight Z-score and the change in weight Z-score between birth and hospital discharge (rs=-0.095, -0.151 respectively, P<0.01), positively correlated with death/withdrawal of care (rs=0.196, P<0.01). CONCLUSIONS: Antibiotics used for VLBW/ELBW infants in NICUs of Hunan Province are obviously prolonged in many NICUs. The proportion of routine use of third-generation of cephalosporins and carbapenems antibiotics is high among the NICUs.

The NLRP3 inflammasome functions as a driver of the myelodysplastic syndrome phenotype.

Despite genetic heterogeneity, myelodysplastic syndromes (MDSs) share features of cytological dysplasia and ineffective hematopoiesis. We report that a hallmark of MDSs is activation of the NLRP3 inflammasome, which drives clonal expansion and pyroptotic cell death. Independent of genotype, MDS hematopoietic stem and progenitor cells (HSPCs) overexpress inflammasome proteins and manifest activated NLRP3 complexes that direct activation of caspase-1, generation of interleukin-1ß (IL-1ß) and IL-18, and pyroptotic cell death. Mechanistically, pyroptosis is triggered by the alarmin S100A9 that is found in excess in MDS HSPCs and bone marrow plasma. Further, like somatic gene mutations, S100A9-induced signaling activates NADPH oxidase (NOX), increasing levels of reactive oxygen species (ROS) that initiate cation influx, cell swelling, and ß-catenin activation. Notably, knockdown of NLRP3 or caspase-1, neutralization of S100A9, and pharmacologic inhibition of NLRP3 or NOX suppress pyroptosis, ROS generation, and nuclear ß-catenin in MDSs and are sufficient to restore effective hematopoiesis. Thus, alarmins and founder gene mutations in MDSs license a common redox-sensitive inflammasome circuit, which suggests new avenues for therapeutic intervention.

TGF-ß-inducible microRNA-183 silences tumor-associated natural killer cells.

Transforming growth factor ß1 (TGF-ß), enriched in the tumor microenvironment and broadly immunosuppressive, inhibits natural killer (NK) cell function by yet-unknown mechanisms. Here we show that TGF-ß-treated human NK cells exhibit reduced tumor cytolysis and abrogated perforin polarization to the immune synapse. This result was accompanied by loss of surface expression of activating killer Ig-like receptor 2DS4 and NKp44, despite intact cytoplasmic stores of these receptors. Instead, TGF-ß depleted DNAX activating protein 12 kDa (DAP12), which is critical for surface NK receptor stabilization and downstream signal transduction. Mechanistic analysis revealed that TGF-ß induced microRNA (miR)-183 to repress DAP12 transcription/translation. This pathway was confirmed with luciferase reporter constructs bearing the DAP12 3' untranslated region as well as in human NK cells by use of sense and antisense miR-183. Moreover, we documented reduced DAP12 expression in tumor-associated NK cells in lung cancer patients, illustrating this pathway to be consistently perturbed in the human tumor microenvironment.

[Protective Effect of Lycium ruthenicum on Peripheral Blood System Against Radiation Injury in Mice].

OBJECTIVE: To study the protective effect of Lycium ruthenicum on peripheral blood system against radiation injury in mice. METHODS: Kunming mice were randomly divided into control group, model group, positive group and Lycium ruthenicum high dose (8 g/kg), middle dose (4 g/kg) and low dose (2 g/kg)treatment groups that experimented three days after irradiation. In the same way, groups were set at 7 days and 14 days after irradiation respectively. Lycium ruthenicum extract were administered orally to the mice in the three Lycium ruthenicum treatment groups and normal saline were administered orally to the mice in control group and model group for 14 days. Positive group were treated with radioprotective agent amifostine (WR-2721) at 30 min before irradiation. Except control group, mice in other groups received quantity of 5 Gy X-radiation whole body evenly with one time. Hemogram, organ index, DNA, Caspase-3, Caspase-6 and P53 contents were observed at the 3rd, 7th and 14th day after irradiation. RESULTS: Lycium ruthenicum significantly increased the total red blood cell count, hemoglobin count, the indexes of spleen and thymus and bone marrow DNA contents (P < 0.05), as well as decreased Caspase-3 and Caspase-6 contents in serum and the expression of P53 in intestinal crypt epithelial cells. CONCLUSION: The results showed that Lycium ruthenicum had protective effects on peripheral blood system against radiation injury in mice.

A Protective Layer of UIO-66/Reduced Graphene Oxide to Stabilize Zinc-Metal Anodes toward High-Performance Aqueous Zinc-Ion Batteries.

Rechargeable aqueous Zn-ion batteries with a Zn anode hold great promise as promising candidates for advanced energy storage systems. The construction of protective layer coatings on Zn anode is an effective way to suppress the growth of Zn dendrites and water-induced side reactions. Herein, we reported a series of UIO-66 materials with different concentrations of reduced graphene oxide (rG) coated onto the surface of Zn foil (Zn@UIO-66/rGx; x = 0.05, 0.1, and 0.2). Benefiting from the synergistic effect of UIO-66 and rG, symmetric cells with Zn@UIO-66/rGx (x = 0.1) electrodes exhibit excellent reversibility (e.g., long cycling life over 1100 h at 1 mA cm-2/1 mAh cm-2) and superior rate capability (e.g., over 1100 and 400 h at 5 mA cm-2/2.5 mAh cm-2 and 10 mA cm-2/5 mAh cm-2, respectively). When the Zn@UIO-66/rG0.1 anode was paired with the NaV3O8·1.5H2O (NVO) cathode, the Zn@UIO-66/rG0.1||NVO cell also delivered a high reversible capacity of 189.9 mAh g-1 with an initial capacity retention of 61.3% after 500 cycles at 1 A g-1, compared to the bare Zn||NVO cell with only 92 cycles.

From graphite of used lithium-ion batteries to holey graphite coated by carbon with enhanced lithium storage capability.

The efficient recycling of waste graphite anode from used lithium-ion batteries (LIBs) has attracted considerable concerns mainly owing to the environment protection and reutilization of resources. Herein, we reported a rational and facile strategy for the synthesis of holey graphite coated by carbon (hG0.01@C0.10) through the separation, purification and creation of holey structures of waste graphite by using NaOH and carbon-coating by using phenolic resin. The holey structures facilitate the hG0.01@C0.10 with the quick penetration of electrolytes and rapid diffusion of Li+. The carbon coating is more favorable for hG0.01@C0.10 with improved electronic conductivity and less alleviated volume during the cycles. Benefiting from the synergistic effect of holey structures and carbon coating, the hG0.01@C0.10 as anode for LIBs displays a high reversible capacity of 377.6 mAh g-1 at 0.5 C and superior rate capabilities (e.g., 348.0 and 274.7 mAh g-1 at 1 and 2 C, respectively) and maintains a high reversible capacity of 278.7 mAh g-1 at 1 C after 300 cycles with an initial capacity retention of 80.0 %.

Icariside II induces apoptosis of melanoma cells through the downregulation of survival pathways.

This study evaluated the antitumor effects of icariside II (IS), isolated from Herba Epimedii, on in vitro and in vivo models of melanoma and determined its mechanism of apoptosis. Mouse (B16) and human (A375, SK-MEL-5) melanoma cell lines were treated with IS at different concentrations (0-100 µM). Cell viability and proliferation was detected by WST-1 assay and with the xCELLigence system, respectively. Apoptosis was measured by the annexin-V/PI flow cytometric assay. Western blot was used to measure cleaved caspase 3, survivin, P-STAT3, P-ERK and P-AKT. B16 and A375 cells were injected subcutaneously into C57BL/6J and BALB/c-nu mice, respectively. After 1 wk, IS solution at (50 mg/kg, 100 mg/kg) was administered by intraperitoneal injection 3 times for a week. Tumor size was measured with an electronic digital caliper. IS inhibited the proliferation of melanoma cells in a dose- and time-dependent manner. Treatment of A375 cells with IS resulted in an increased number of apoptotic cells ranging from 5.6% to 26.3% mirrored by increases in cleaved caspase-3 and a decrease in survivin expression. IS significantly inhibited the activation of the JAK-STAT3 and MAPK pathways but promoted an unsustained activation peak of the PI3K-AKT pathway. IS administration (50 mg/kg) resulted in a 47.5% decreased tumor volume in A375 bearing mice. Furthermore, IS administration (50 mg/kg, 100 mg/kg) resulted in 41% and 49% decreased tumor volume in B16 bearing mice, respectively. IS dramatically inhibited the proliferation of melanoma cells in vivo and in vitro through the regulation of apoptosis. These effects demonstrate the ability of IS to effectively overcome the survival signals of tumor cells, which support further preclinical evaluation of IS in cancer as a new potential chemotherapeutic agent.

An efficient electrolyte additive of 1,3,6-hexanetricarbonitrile for high performance aqueous zinc-ion batteries.

The growth of Zn dendrites and parasitic side reactions between electrode and electrolyte are major obstacles to the development of rechargeable aqueous zinc-ion batteries. To address these critical issues, the use of nitrile organic compounds as electrolyte additives holds great promise. Herein, for the first time, we prepared a small volume concentration (x) of 1,3,6-Hexanetricarbonitrile (HTCN-x) as additives into zinc trifluoromethanesulphonate (Zn(OTF)2) electrolyte and studied their electrochemical properties in Zn||ZnxV2O5·nH2O (Zn||ZVO) cells. It was found that the strong interaction between H2O and HTCN could significantly reduce the population of solvated H2O outside the solvation sheath, leading to reduced side reactions in the aqueous Zn(OTF)2 electrolyte. Moreover, the HTCN additive also facilitates the formation of strong and stable solid electrolyte interphase (SEI) film on the surface of the Zn anode, which effectively prevents the growth of Zn dendrites and the anode corrosion caused by the electrolyte. As a result, the HTCN-x (x = 0.3) electrolyte enabled the symmetrical Zn||Zn cell to cycle over 950 h at a current of 1 mA cm-2 with a limited capacity of 1 mAh cm-2. When the HTCN-0.3 electrolyte was used in Zn||ZVO cell, the cell delivered a high initial capacity of 355.6 mAh g-1 at 0.1 A g-1 and maintained a high capacity of 330.0 mAh g-1 at 1 A g-1 after 465 cycles.

Salt Anion Amphiphilicity-Activated Electrolyte Cosolvent Selection Strategy toward Durable Zn Metal Anode.

One effective solution to inhibit side reactions and Zn dendrite growth in aqueous Zn-ion batteries is to add a cosolvent into the Zn(CF3SO3)2 electrolyte, which has the potential to form a robust solid electrolyte interface composed of ZnF2 and ZnS. Nevertheless, there is still a lack of discussion on a convenient selection method for cosolvents, which can directly reflect the interactions between solvent and solute to rationally design the electrolyte solvation structure. Herein, logP, where P is the octanol-water partition coefficient, a general parameter to describe the hydrophilicity and lipophilicity of chemicals, is proposed as a standard for selecting cosolvents for Zn(CF3SO3)2 electrolyte, which is demonstrated by testing seven different types of solvents. The solvent with a logP value similar to that of the salt anion CF3SO3- can interact with CF3SO3-, Zn2+, and H2O, leading to a reconstruction of the electrolyte solvation structure. To prove the concept, methyl acetate (MA) is demonstrated as an example due to its similar logP value to that of CF3SO3-. Both the experimental and theoretical results illustrate that MA molecules not only enter into the solvation shell of CF3SO3- but also coordinate with Zn2+ or H2O, forming an MA and CF3SO3- involved core-shell solvation structure. The special solvation structure reduces H2O activity and contributes to forming an anion-induced ZnCO3-ZnF2-rich solid electrolyte interface. As a result, the Zn||Zn cell and Zn||NaV3O8·1.5H2O cell with MA-involved electrolyte exhibit superior performances to that with the MA-free electrolyte. This work provides an insight into electrolyte design via salt anion chemistry for high-performance Zn batteries.

Bcl11b sustains multipotency and restricts effector programs of intestinal-resident memory CD8+ T cells.

The networks of transcription factors (TFs) that control intestinal-resident memory CD8+ T (TRM) cells, including multipotency and effector programs, are poorly understood. In this work, we investigated the role of the TF Bcl11b in TRM cells during infection with Listeria monocytogenes using mice with post-activation, conditional deletion of Bcl11b in CD8+ T cells. Conditional deletion of Bcl11b resulted in increased numbers of intestinal TRM cells and their precursors as well as decreased splenic effector and circulating memory cells and precursors. Loss of circulating memory cells was in part due to increased intestinal homing of Bcl11b-/- circulating precursors, with no major alterations in their programs. Bcl11b-/- TRM cells had altered transcriptional programs, with diminished expression of multipotent/multifunctional (MP/MF) program genes, including Tcf7, and up-regulation of the effector program genes, including Prdm1. Bcl11b also limits the expression of Ahr, another TF with a role in intestinal CD8+ TRM cell differentiation. Deregulation of TRM programs translated into a poor recall response despite TRM cell accumulation in the intestine. Reduced expression of MP/MF program genes in Bcl11b-/- TRM cells was linked to decreased chromatin accessibility and a reduction in activating histone marks at these loci. In contrast, the effector program genes displayed increased activating epigenetic status. These findings demonstrate that Bcl11b is a frontrunner in the tissue residency program of intestinal memory cells upstream of Tcf1 and Blimp1, promoting multipotency and restricting the effector program.

CX3CR1 deficiency-induced TIL tumor restriction as a novel addition for CAR-T design in solid malignancies.

Advances in the understanding of the tumor microenvironment have led to development of immunotherapeutic strategies, such as chimeric antigen receptor T cells (CAR-Ts). However, despite success in blood malignancies, CAR-T therapies in solid tumors have been hampered by their restricted infiltration. Here, we used our understanding of early cytotoxic lymphocyte infiltration of human lymphocytes in solid tumors in vivo to investigate the receptors in normal, adjacent, and tumor tissues of primary non-small-cell lung cancer specimens. We found that CX3CL1-CX3CR1 reduction restricts cytotoxic cells from the solid-tumor bed, contributing to tumor escape. Based on this, we designed a CAR-T construct using the well-established natural killer group 2, member D (NKG2D) CAR-T expression together with overexpression of CX3CR1 to promote their infiltration. These CAR-Ts infiltrate tumors at higher rates than control-activated T cells or IL-15-overexpressing NKG2D CAR-Ts. This construct also had similar functionality in a liver-cancer model, demonstrating potential efficacy in other solid malignancies.

Positive regulatory domain I (PRDM1) and IRF8/PU.1 counter-regulate MHC class II transactivator (CIITA) expression during dendritic cell maturation.

Dendritic cells (DCs) are key mediators of immune function through robust and tightly regulated presentation of antigen in the context of the MHC Class II. MHC Class II expression is controlled by the transactivator CIITA. CIITA expression in conventional DCs is uniquely dependent on an uncharacterized myeloid cell-specific promoter, CIITApI. We now identify in vivo the promoter structure and factors regulating CIITApI. In immature DCs transcription requires binding of PU.1, IRF8, NFκB, and Sp1 to the promoter. PU.1 binds independently at one site and in a required heterodimer with IRF8 at a composite element. DCs from IRF8-null mice have an unoccupied CIITApI promoter that can be rescued by reconstitution with IRF8 in vitro. Furthermore, mutation of either PU.1 site or the IFR8 site inhibits transcriptional activation. In vivo footprinting and chromatin immunoprecipitation reveals that DC maturation induces complete disassociation of the bound activators paralleled by recruitment of PRDM1/Blimp-1 to the promoter. PRDM1 is a transcriptional repressor with essential roles in B cells, T cells, NK cells, and DCs. We show that PRDM1 co-repressors, G9a and HDAC2, are recruited to CIITApI, leading to a loss of histone acetylation and acquisition of histone H3K9 dimethylation and heterochromatin protein 1γ (HP1γ). PRDM1 binding also blocks IRF8-mediated activation dependent on the PU.1/IRF composite element. Together these findings reveal the mechanisms regulating CIITA and, thus, antigen presentation in DCs, demonstrating that PRDM1 and IRF8/PU.1 counter-regulate expression. The activity of PRDM1 in silencing all three cell type-specific CIITA promoters places it as a central regulator of antigen presentation.

A critical role for phosphatase haplodeficiency in the selective suppression of deletion 5q MDS by lenalidomide.

Lenalidomide is the first karyotype-selective therapeutic approved for the treatment of myelodysplastic syndromes (MDS) owing to high rates of erythroid and cytogenetic response in patients with chromosome 5q deletion [del(5q)]. Although haploinsufficiency for the RPS14 gene and others encoded within the common deleted region (CDR) have been implicated in the pathogenesis of the del(5q) phenotype, the molecular basis of the karyotype specificity of lenalidomide remains unexplained. We focused our analysis on possible haplodeficient enzymatic targets encoded within the CDR that play key roles in cell-cycle regulation. We show that the dual specificity phosphatases, Cdc25C and PP2Acalpha, which are coregulators of the G(2)-M checkpoint, are inhibited by lenalidomide. Gene expression was lower in MDS and acute myeloid leukemia (AML) specimens with del(5q) compared with those with alternate karyotypes. Lenalidomide inhibited phosphatase activity either directly (Cdc25C) or indirectly (PP2A) with corresponding retention of inhibitory phospho-tyrosine residues. Treatment of del(5q) AML cells with lenalidomide induced G(2) arrest and apoptosis, whereas there was no effect in nondel(5q) AML cells. Small interfering RNA (shRNA) suppression of Cdc25C and PP2Acalpha gene expression recapitulated del(5q) susceptibility to lenalidomide with induction of G(2) arrest and apoptosis in both U937 and primary nondel(5q) MDS cells. These data establish a role for allelic haplodeficiency of the lenalidomide inhibitable Cdc25C and PP2Acalpha phosphatases in the selective drug sensitivity of del(5q) MDS.

Different diagnostic power of anti-Mullerian hormone in evaluating women with polycystic ovaries with and without hyperandrogenism.

PURPOSE: The aim of this study is to compare the secretory profiles and diagnostic power of anti-Mullerian hormone (AMH) for the PCOS patient with and without hyperandrogenism. METHODS: One hundred and thirty-one PCOS patients with oligomenorrhea or amenorrhea were recruited into the study. Sixty-two and sixty-nine patients had and did not have hyperandrogenism (HA+) hyperandrogenism (HA-), respectively. Sera were collected for determining the levels of AMH, basal sexual hormones, glucose and lipid metabolic indicators. RESULTS: The AMH serum levels of PCOS patients were significantly higher than the control group, with the highest AMH serum level in the HA+ group. The cut-off value for predicting PCOS patients of all types was 3.92 ng/mL, with a sensitivity of 65 %, and specificity of 62 %. The cut-off value for predicting PCOS patients in the HA+ group was 4.23 ng/mL, with a sensitivity of 82 %, and specificity of 64 %. The cut-off value for predicting PCOS patients in the HA- group was 3.76 ng/mL, with a sensitivity of 64 %, and specificity of 62 %. In the HA+ group, AMH was negatively associated with FSH and positively associated with LH. In the HA- group, AMH was negatively associated with HDL and positively associated with BMI, fasting glucose and LDL. CONCLUSIONS: AMH is only suitable for predicting the PCOS patients with hyperandrogenism. The diagnostic power of AMH is limited when used to predict patients without hyperandrogenism. It reflects the differences in pathophysiology and severity of disrupted folliculogenesis between the two subtypes.

[Expression and significance of Twist1 and MMP-2 in endometrial endometrioid adenocarcinoma].

OBJECTIVE: To investigate the expression of Twist1 and MMP-2 protein and their significance in endometrial endometrioid adenocarcinoma. METHODS: The expression of Twist1 and MMP-2 protein in 70 cases of endometrial endometrioid adenocarcinoma was detected on tissue chips using immunohistochemical staining. RESULTS: The positive rates of Twist1 and MMP-2 protein expression were 65.7% and 67.1%, respectively. Both of the high expressions of Twist1 and MMP-2 were positively correlated with FIGO staging and tumor myometrial invasion (P < 0.05, respectively). Also the high expression of Twist1 was positively correlated with ovarian metastasis and the expression of MMP-2 was positively correlated with tumor grading (P < 0.05). The patients' overall survival and relapse-free survival in the group of high Twist1 expression were shorter than that in the group of low Twist1 expression (P < 0.05). The expression of MMP-2 was positively correlated with Twist1 expression (P < 0.01, respectively). CONCLUSIONS: The expression of Twist1 may be closely correlated with the tumor invasion, metastasis and prognosis in patients with endometrial endometrioid adenocarcinoma. The expression of Twist1 has a close relationship with MMP-2 in endometrial endometrioid adenocarcinoma.