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This study investigated a 2-stage arterial switch operation (ASO) to treat transposition of the great arteries (TGA) with intact ventricular septum (TGA-IVS) in late referral patients.We retrospectively analyzed patients with TGA-IVS or TGA with restricted ventricular septal defects who had undergone 2-stage ASO at our institution from February 2007 to August 2018. Included were 41 patients: 21 (51.2%) who had undergone long-term 2-stage ASO and 20 (48.8%) who had undergone rapid 2-stage ASO.The long-term 2-stage group was older at ASO (3.5 vs 25 months; P.001). Results were more satisfactory in the long-term group than in the rapid group for intensive care unit time (P = .004), mechanical ventilation time (P = .004), and length of stay (P = .007). No in-hospital death occurred in the long-term group, and the postoperative course was more manageable in the long-term group than in the rapid group. However, the risk of significant neoaortic regurgitation was lower in the rapid group, which also had a better left ventricular ejection fraction.The long-term group achieved better early-term outcomes than the rapid group. However, a high risk of neoaortic regurgitation and myocardial dysfunction was also noted.

Vascular endothelial growth factor A (VEGFA) plays an important role in the healing of skin wound. However, the application of VEGFA protein in clinic is limited because of its high cost manufacturing, complicated purification and poor pharmacokinetic profile. Herein, we developed nucleoside-modified mRNA encoding VEGFA encapsulated ionizable lipid nanoparticles (LNP) to improve angiogenesis and increase wound healing rate. First, VEGFA mRNA was synthesized by an in vitro transcription (IVT) method. After that, VEGFA mRNA-LNP was prepared by encapsulating mRNA in ionizable lipid based nanoparticles via a microfluidic mixer. The physicochemical properties of VEGFA mRNA-LNP were investigated via dynamic light scattering (DLS) and transmission electron microscopy (TEM). The results showed that the VEGFA mRNA-LNP possessed regular spherical morphology with an average size of 112.67 nm and a negative Zeta potential of -3.43 mV. The LNP delivery system had excellent lysosome escape capability and high transfection efficiency. ELISA and Western Blot analysis indicated that the mRNA-LNP could express VEGFA protein in Human umbilical vein endothelial cells (HUVECs). Besides, endothelial tube formation, cell proliferation and scratch assays were performed. The results revealed VEGFA mRNA-LNP boosted angiogenesis, cell proliferation and cell migration by expressing VEGFA protein. Finally, C57BL/6 mouse model of skin wound was established and intradermally treated with VEGFA mRNA-LNP. The VEGFA mRNA-LNP treated wounds were almost healed with an average wound size of 1.56 mm

Accessing freshwater resources for agriculture becomes more complex due to increasing demands and declining water quality. Alternative water sources, such as saline water, require ad hoc solutions. Therefore, understanding roots' response to saline water is crucial for future agriculture. We examined the response of three grapevine rootstocks (Paulsen 1103, Richter 110 and SO4) to salt stress. The rootstocks were subjected to two salinity treatments: 10 mM and 30 mM NaCl (EC = 2 and 4 ds/m, respectively). Root and shoot samples were taken at the end of the experiment for morphologic and ionomic analyses. The specific root area (SRA) increased in response to salinity for all three rootstocks due to root tissue density and average root diameter reductions. Salinity also led to increased root Na

Protein kinase N3 (PKN3), an AGC-family member, is often overexpressed in breast tumor cells. RNAi therapy is a promising approach to inhibit tumor growth by reducing the expression of PKN3. In this report, lipid nanoparticles encapsulated with new shRNA PKN3 (SS-LNP/shPKN3) with redox-responsiveness were developed in order to specifically down-regulate the expression of PKN3 for breast cancer treatment. The SS-LNP/shPKN3 was prepared by microfluidic method using disulfide bonds based ionizable lipid as main component. The as-prepared SS-LNP/shPKN3 lipid nanoparticles were characterized via using dynamic light scattering (DLS) and transmission electron microscopy (TEM). The results indicated that the obtained SS-LNP/shPKN3 exhibited uniform particle size and regular spherical morphology. Moreover, glutathione (GSH) triggered release of shPKN3 confirmed the redox-responsiveness of the SS-LNP/shPKN3. Finally, the anti-tumor effect of SS-LNP/shPKN3 was evaluated against MDA-MB-231 cells and derived xenograft tumor bearing mice. It was found that the SS-LNP/shPKN3-2 had the highest PKN3 protein inhibition rate of 60.8% and tumor inhibition rate of 62.3%. Taken together, the SS-LNP/shPKN3 might be a potential therapeutic strategy for breast cancer.

Rare-earth transition metal ultra-thin TbFeCo amorphous films in the thickness range from 3.5 to 35 nm are radio frequency (RF)-sputtered based on a composition target method. Perpendicular magnetic anisotropy (PMA) is found to be strongly related to the thickness of sputtered TbFeCo film in this thickness range. No PMA can be observed in the 3.5 nm film, which may be too short to form local intrinsic single ion anisotropy. Strong PMA develops with the increasing film thickness. A critical compensation thickness of ~14 nm is found, where both the largest coercivity and lowest saturation magnetization are demonstrated. Across this critical thickness the change in polarity of the EHE loops is observed, corresponding to a transition from a FeCo-dominant to a Tb-dominant configuration. Additionally, the magnetic properties of the alloy films are shown to be significantly affected by the substrates used for deposition, which can be mainly attributed to preferential oxidization of the rare-earth Tb element in the films at the interfaces between the film and the substrates. Our results provide a simple method to manipulate magnetic properties of ultra-thin TbFeCo alloy films for the applications in spintronic devices.

Dimeric artesunate phospholipid (ART-GPC), an amphiphilic derivative of artemisinin dimer reported in our previous work, can be applied to treat malaria effectively. The objective of this study is to develop a facile method for the industrial production of ART-GPC liposomes. Conventional methods including thin film hydration (TFH), ethanol injection (EI), and freeze drying (FD) were used to prepare ART-GPC liposomes, and the resultants presented poor physicochemical properties. Fortunately, a modified thin film hydration method (MTFH) by forming thin film of ART-GPC composed of fine lipid bilayer structure in the vials showed promise for the liposomes production. A quality design strategy (solvents, pressure, hydration time, and temperature) was performed to obtain optimal physicochemical characteristics and production conditions. Thereafter, ART-GPC liposomes are produced under GMP conditions with the size of 176.32 nm, PDI of 0.17, zeta potential of -25.79 mV, and osmotic pressure of 297.33 mOsm/kg, confirming the scalability and reproductivity of the MTFH technology. It is the first report that the MTFH method allows liposomes to be preserved in a dry film state and in-situ hydrated in injection vials with excellent performance. Conclusively, the MTFH method is a promising technology for the large-scale production of ART-GPC liposomes.

Sb2Se3, as a new class of functional materials, has recently emerged for solar energy conversion owing to its low cost and high activity. In this paper, a facile electrodeposition method allows uniform formation of vertically aligned Sb2Se3 nanorods arrays. The physical and chemical properties of Sb2Se3 films can be modulated by varying the used substrates. A more uniform nanorod-shaped Sb2Se3(Se) film is developed on the Se-coated Ni substrate with better crystallinity, improved light absorption ability, suitable band structure, efficient photogenerated carriers transport. Additionally, the Sb2Se3(Se) film exhibits the better photocatalytic activity, and degrades 74.7% methylene blue (MB) after irradiation for 60 ​min, as compared to the Sb2Se3(Ni) film. The research provides an effective approach for synthesizing well-aligned Sb2Se3 nanorods arrays, and reveals a new direction for designing high-efficiency photocatalysis.

Co2FeAl (CFA) films in a wide thickness range between 2 and 100 nm are sputtered at room temperature. Perpendicular magnetic anisotropy (PMA) is achieved in the annealed structure of Pd/CFA/MgO with CFA thickness ranging between 2.3 and 4.9 nm. PMA as high as 2 × 106 erg/cm3 is demonstrated in the structures annealed in the temperature range between 300 and 350 °C. Positive contributions to the PMA made by the interfaces of Pd/CFA and CFA/MgO are identified. For the as-deposited structure of MgO/CFA/Ta with thick CFA alloy up to 5 nm or above a high effective saturation magnetization of 983.9 ± 30.1 emu/cc is derived from the fitting and an in-plane uniaxial magnetic anisotropy of 104 erg/cm3 in magnitude is revealed by angular dependent magnetic measurements. In addition to the increase in saturation magnetization, a fourfold cubic magnetic anisotropy is found to develop with annealing, in line with the improvement of the crystalline structure confirmed by X-ray diffraction measurements. Out results provide some useful information for the design of the CFA-based magnetoelectronic devices.

In recent years, Te films are an active semiconductor photocatalysts because of the suitable photoelectrochemical properties. In this paper, Te films were successfully fabricated via a facile electrodeposition method with the addition of NaCl into electrolytic solution and assessed for photocatalytic application. The physicochemical performances of Te films were characterized via many analytical methods. It was discovered that the resulting Te films can present different physicochemical properties controllably depending on the NaCl used. Photodegradation experimental result indicates that the chip-shaped Te films show superior photocatalytic activity towards MB degradation. The degradation efficiency of 95.1% was achieved in 60 ​min. In addition, the developed films also presented superior stability without an apparent inactivation on photocatalytic activity after four photocatalytic cycles. The markedly enhanced photocatalytic activity of the chip-shaped Te film can be due to the formation of unique structure which can increase the surface area, promote the absorption capability and improve the separation efficiency of photogenerated charge carriers. Notably, in the degradation process of MB, the •OH and •O2− are the main active radical species according to the free radicals capturing experiments and ESR results. A possible photocatalytic degradation mechanism was also proposed. It is hoped that the chip-shaped Te films have potential desirable applications in photocatalytic-industry. This work also offers a promising strategy to improve the performance of Te films to be high and multiple efficient photocatalyst.

A novel method for the detection of antibiotics is developed by combining surface enhanced Raman scattering (SERS) with molecularly imprinted polymers (MIPs) on membrane materials. In this test, eggshell membrane (ESM) is selected as support material, which shows the advantages of low price, easy preparation and more functional groups comparing with other common synthetic membranes. Firstly, Ag nanoparticles with different morphologies are deposited on the surface of ESM by in-situ reduction, and then imprinted polymer is coated by precipitation polymerization to promote high specific sensitivity. The results present that the sensitivity of SERS imprinted membrane is mainly affected by Ag morphology. Under the optimal conditions, it presents a good linear relationship between SERS intensity and spiramycin (SP) concentration, and the coefficient of determination (R2) is 0.9981. It also exhibits the ability of selective detection and retrievability. It can be recycled at least 10 times. This method provides a cheap, stable and sensitive substrate, which broadens the research field of SERS detection of antibiotics.

Amorphous TbFeCo material with perpendicular magnetic anisotropy is currently attracting more attention for potential applications in spintronic devices and logic memories. We systematically investigate magnetic, structural, thermal, optical and electrical properties of TbFeCo alloy films. It shows out-of-plane easy axis of the films turns into in-plane orientation after annealing. Significant increase in saturation magnetization in the temperature range between 400 and 450 °C is revealed by thermomagnetic measurements. The occurrence of crystallization and oxidation at high temperatures is confirmed by X-ray diffraction measurements. Pronounced changes in optical reflectance and sheet resistance are observed with temperature, in line with structural relaxation and change. The activation barriers for crystallization and oxidation are determined to be 1.01 eV and 0.83 eV, respectively, for FeCo-rich and Tb-rich samples. Better thermal stability against crystallization and oxidation is demonstrated in the FeCo-rich sample than the Tb-rich type. Our results provide some useful information for the alloy used in device fabrication.

Background and aims Myocarditis is inflammation in the heart; its pathogenesis is to be further investigated. Activities of micro RNAs (miR) are associated with immune inflammation. This study tests a hypothesis that miR-98 is involved in the development of myocarditis. Methods BALB/c mice were immunized with cardiac α-myosin heavy chain peptides (MyHC-α) to induce myocarditis. The effects of miR-98 on regulation of interleukin (IL)-10 were assessed by real time RT-PCR. Results Mice immunized with MyHC-α showed myocarditis and lower frequency of IL-10 + B cells (B10 cell) in the hearts. Expression of miR-98 was higher, IL-10 was lower, in B cells isolated from the mouse hearts with myocarditis, which was negatively correlated with each other. Exposure to tumor necrosis factor-α up regulated miR-98 expression in B cells. Over-expression of miR-98 suppressed IL-10 expression in B cells. Blocking miR-98 or adoptively transplanting B10 cells attenuated experimental myocarditis. Conclusions miR-98 suppresses IL-10 expression in B cells in the heart, which plays an important role in myocarditis. MiR-98 may be a therapeutic target in the treatment of myocarditis.

A series of amorphous perpendicular TbFeCo films are sputtered using a composite target. The magnetic properties of the TbFeCo films are found to strongly be influenced by sputtering power. The above-RT magnetic compensation point in the films sputtered at high powers is revealed by thermomagnetic measurements. With increasing the power the compensation point shifts considerably towards higher temperature, while only a slight reduction in the Curie temperature occurs. The coercivity is found to initially increase with the power and then falls, which is in accordance with film composition moving from FeCo-rich to Tb-rich side. This can be well explained by the sputtering yield of Tb, which increases faster than FeCo atoms with the power during the sputtering. Our results show a convenient way to adjust the coercivity of the films by modifying the Tb-to-FeCo ratio using the sputtering power.

Redox flow batteries are a promising technology for large scale energy storage systems. Although some types of the battery have been commercialized, the narrow working voltage ranges in the aqueous electrolytes limit their further applications. Organic solvents have shown to allow the use of a wider range of redox couples and the idea of non-aqueous redox flow batteries has been proposed. However, the low solubilities and poor electrochemical properties including stability and reversibility of the redox couples in the organic electrolytes remain a significant issue. In this work, a series of heteroleptic (mixed ligand) iron(II) complexes based on a combination of 2,2′-bipyridine, 1,10-phenanthroline and 4,4′-dimethyl-2,2′-bipyridine ligands were synthesized and their chemical and electrochemical properties investigated. For Fe (II) complexes, it was confirmed that ligand equilibration occurs in solution. The electrochemical properties of the complexes were studied by cyclic voltammogram and chronoamperometric methods. The mechanism of the redox processes of these complexes are proposed and how the mixed ligands will affect the redox potentials of the Fe (II) complexes is discussed. Furthermore, we show that the mixed ligand system can improve the total solubility of redox active species. The mixed ligand approach will also be beneficial for designing other new high solubility redox materials for non- aqueous flow batteries.

CoFeAlSi alloy films are sputtered at room temperature and annealed at various temperatures. X-ray diffraction measurements reveal as-deposited films are amorphous and a nanocrystalline structure is formed after annealing. The coercivity is found to rapidly increase with the annealing temperature, in line with the grown nanocrystalline grain size. The angle dependent magnetic measurements reveal a thermal-induced cubic structural anisotropy develops with the annealing temperature and is superimposed on a growth-induced uniaxial anisotropy with parallel easy axes. Magnetic properties of the alloy are shown to be strongly influenced by the structural change induced by the thermal treatment. The interplay between thermal dependent magnetization and crystallization behavior of the alloy film is revealed by thermomagnetic measurements. A activation barrier of 1.32 eV is determined using the optical reflectance measurements with variable heating rates. Keywords: Metallic alloys, Magnetic materials, Magnetic properties, Thermal properties

CoFeB alloy films have been attracting much research interest for the applications in magnetoelectronic devices. In this study, crystallization and high temperature functional properties in the sputtered CoFeB alloy films were investigated. The as-deposited alloy films were amorphous and a nanocrystalline structure was formed after annealing, confirmed by X-ray measurements. Large variation in saturation magnetization with temperature was observed in thermomagnetic measurements, reflecting the interplay between crystallization and temperature dependent magnetization in the ferromagnetic alloy film. Pronounced changes in electrical resistivity and optical reflectance with temperature were observed in the alloy film, in accompany with the structural change. The activation barrier for crystallization was determined to be 1.32 eV from the electrical measurements, showing a good thermal stability of the alloy film.

Alnico 8 ribbons were prepared by melt-spinning and annealing, and the effects of melt-spinning speeds on the structure and magnetic properties were investigated. The results show that all the as-spun ribbons are composed of the α-Fe(Co)-type α1 and the AlNi-type α2. With speed reducing from 40 m/s to 15 m/s, the preferred orientation enhanced, the grain size and the α2 content increased. The as-spun ribbons show the weak hard magnetic properties. Annealing improves the spinodal structure consisting of the strip-like α1 and α2. The annealed ribbons melt-spun at 15 m/s achieve the optimal magnetic properties with the coercivity of 1558.3 Oe and the maximum magnetic polarisation density of 11.3 kGs, which are codetermined by the shape anisotropy of the strip-like α1 phase and the pinning mechanism of the α2 phase to the α1 domains.

The H3 histone variant CENP-A is an epigenetic marker critical for the centromere identity and function. However, the precise regulation of the spatiotemporal deposition and propagation of CENP-A at centromeres during the cell cycle is still poorly understood. Here, we show that CENP-A is phosphorylated at Ser68 during early mitosis by Cdk1. Our results demonstrate that phosphorylation of Ser68 eliminates the binding of CENP-A to the assembly factor HJURP, thus preventing the premature loading of CENP-A to the centromere prior to mitotic exit. Because Cdk1 activity is at its minimum at the mitotic exit, the ratio of Cdk1/PP1α activity changes in favor of Ser68 dephosphorylation, thus making CENP-A available for centromeric deposition by HJURP. Thus, we reveal that dynamic phosphorylation of CENP-A Ser68 orchestrates the spatiotemporal assembly of newly synthesized CENP-A at active centromeres during the cell cycle.

Objective The aim of this review was to evaluate the feasibility, safety, and potential benefits of single-incision laparoscopic splenectomy (SILS-Sp). Methods We conducted a systemic review of literature between 2009 and 2012 to retrieve all relevant articles. Results A total of 29 studies with 105 patients undergoing SILS-Sp were reviewed. Fifteen studies used a commercially available single-port device. The range of body mass index was 14.7–41.4 kg/m2. Six studies described combined operations including cholecystectomy (n = 8), mesh-pexy (n = 1), and pericardial devascularizaion (n = 1). The ranges of operative times and estimated blood losses were 28–420 min and 0–350 mL, respectively. Of 105 patients, three patients (2.9%) required additional ports, two patients (1.9%) were converted to open, and three patients (2.9%) to conventional multiport laparoscopic splenectomy (overall conversion rate, 4.8%). Postoperative bleeding occurred in two patients (1.9%) who both required reoperation. Overall mortality was 0% (0/105). The length of postoperative stay varied across reports (1–11 d). Among four comparative studies, one showed greater estimated blood loss and lower numeric pain rating scale score in the SILS-Sp group than in the multiport laparoscopic splenectomy group (206.25 ± 142.45 versus 111.11 ± 99.58 mL) and (3.81 ± 0.91 versus 4.56 ± 1.29), respectively. Another comparative study showed that SILS-Sp was associated with a shorter operative time (92.5 versus 172 min; P = 0.003), lower conversion rate, equivalent length of hospital stay, reduced mortality, similar morbidity, and comparable postoperative narcotic requirements. Conclusions In early series of highly selected patients, SILS-Sp appears to be feasible and safe when performed by experienced laparoscopic surgeons. However, as an emerging operation, publication bias is a factor that should be considered before we can draw an objective conclusion.

Ferriferrous oxide/ l -cysteine (denoted as Fe 3 O 4 /Cys) magnetic microspheres were prepared via a solvothermal route. The content of thiol groups of Fe 3 O 4 /Cys microspheres was determined by a DTNB method, and the maximum content of thiol group was as high as 653.62 μmol/g. The Fe 3 O 4 /Cys microspheres have strong ability to adsorb Pb (II) in aqueous solution and the maximum adsorption capacity reaches 459.33 mg/g. The data at adsorption equilibrium are best fitted with a Langmuir model. In addition, the Fe 3 O 4 /Cys adsorbent can be easily enriched by a magnetic field, and it indicates that as-synthesized adsorbent can be a promising regenerated adsorbent for Pb (II) ions.

Almost 100% of APL patients carry chimeric transcripts encoding truncated RARalpha fused to homo-oligomerization domains from partner proteins. To gain further insights into the cellular transformation mechanisms mediated by RARalpha fusion proteins, thorough structure/function analyses have been performed and identified the POZ homo-oligomerization domain as the minimal transformation domain that is necessary and sufficient for PLZF-RARalpha-mediated in vitro transformation of primary hematopoietic cells. A transformation-incompetent PLZF-RARalpha mutant defective in homo-oligomerization but not corepressor interaction could be rescued by synthetic FKBP-oligomerization domains. Furthermore, an artificial FKBP-RARalpha construct not only mimicked various biochemical properties of bona fide RARalpha fusion proteins but also mediated an ATRA-dependent transformation. Taken together, these findings endorse an oligomerization-dependent mechanism for RARalpha-mediated transformation and suggest a potential avenue for molecular therapy.

The thin sectioning has been widely applied in electron microscopy (EM), and successfully used for an in situ observation of inner ultrastructure of cells. This powerful technique has recently been extended to the research field of atomic force microscopy (AFM). However, there have been no reports describing AFM imaging of serial thin sections and three-dimensional (3-D) reconstruction of cells and their inner structures. In the present study, we used AFM to scan serial thin sections approximately 60 nm thick of a mouse embryonic stem (ES) cell, and to observe the in situ inner ultrastructure including cell membrane, cytoplasm, mitochondria, nucleus membrane, and linear chromatin. The high-magnification AFM imaging of single mitochondria clearly demonstrated the outer membrane, inner boundary membrane and cristal membrane of mitochondria in the cellular compartment. Importantly, AFM imaging on six serial thin sections of a single mouse ES cell showed that mitochondria underwent sequential changes in the number, morphology and distribution. These nanoscale images allowed us to perform 3-D surface reconstruction of interested interior structures in cells. Based on the serial in situ images, 3-D models of morphological characteristics, numbers and distributions of interior structures of the single ES cells were validated and reconstructed. Our results suggest that the combined AFM and serial-thin-section technique is useful for the nanoscale imaging and 3-D reconstruction of single cells and their inner structures. This technique may facilitate studies of proliferating and differentiating stages of stem cells or somatic cells at a nanoscale.

We describe here two mouse mutants, yellow submarine (Ysb) and light coat and circling (Lcc). Ysb arose as the result of insertions of a transgene, pAA2, into the genome. Lcc is an independent, radiation-induced mutation. Both mutants are characterized by recessive circling behavior and deafness, associated with a non-segregating, semi-dominant yellow coat color. Complementation tests showed that Ysb and Lcc are allelic. We attribute the yellow coat in Ysb and Lcc mice to the absence of black awl overhairs, increased agouti zigzag underhairs, and the presence of agouti awls with long subapical yellow pigment. Chromosomal mapping and genomic characterization showed the Ysb and Lcc mutations involve complex chromosomal rearrangements in overlapping regions of mouse chromosome 3, A2/A3-B/C and B-E1, respectively. Ysb and Lcc show for the first time, to our knowledge, the presence of genes in the B-C region of chromosome 3 important for balance and hearing and the pigmentation and specification of coat hair.

The -93 SNP of MLH1 gene is associated with MLH1 gene methylation in endometrial and colorectal cancers. We undertook luciferase reporter assay and electrophoretic mobility shift assay (EMSA) to test whether the -93 SNP affects the MLH1 gene expression. The luciferase activity for -93A plasmid is significantly lower than -93G plasmid. In EMSA experiments, the -93A and -93G probes have different binding affinity to nuclear proteins of JEG3 cells. Our data indicate that -93 SNP affects MLH1 gene expression by altering protein binding to the promoter of MLH1 gene.

The TAL-1 gene is located on chromosome 1p32. In about 20% of T-cell acute lymphoblastic leukemias (T-ALL), this gene is disrupted in its 5' portion by a site-specific 100-kg deletion and is fused with the 5' part of the SIL gene, to form SIL-TAL-1 chimeric gene. In this study, we established a "nested" retrotranscriptase/polymerase chain reaction (RT/PCR) technique which allows detection of the SIL-TAL-1 transcriptional expression. A chimeric mRNA was observed in four of 17 T-ALL cases and has been shown to result from the fusion between the exon 1 of SIL and exon 3 of TAL. A sensitivity test showed that this RT/PCR procedure could detect one leukemic cell among 10(6) normal cells. A positive RT/PCR result was obtained in two cases during clinical remission, suggesting the presence of minimal residual disease (MRD). One patient developed clinical relapse 3 months after PCR positivity. Moreover, analysis of the Tald rearrangement by DNA-based PCR in four patients with SIL-TAL-1 fusion revealed the type A (Tald1) rearrangement in all cases. Sequence analysis demonstrated the presence of N region and non-random "P" nucleotide, as well as base deletions at the genomic SIL-TAL-1 joining site. These data indicate that detection of TAL-1 gene abnormality is important for diagnosis and monitoring of MRD in a subset of T-ALL.

1022 The paired box domain of PAX5 was reported to fuse with the sequence of promylocytic leukemia (PML) to produce PAX5-PML chimeric protein in two patients with B-cell acute lymphoblastic leukemia (B-ALL). In the present studies, we found, by gel-shift assays, that PAX5-PML bound to a panel of PAX5-consensus sequence as a homodimer with reduction of its DNA-binding affinities in comparison with wild-type PAX5. In transient transfection assays using 293T and HeLa cells, and retrovirus transduction of murine hematopoietic stem/progenitor cells together with quantitative real-time polymerase chain reaction (RQ-PCR) analysis, PAX5-PML inhibited wild-type PAX5 target gene transcriptional activity. Studies comparing PAX5-PML with PAX5-PML(ΔCC) demonstrated that the coiled-coil protein interaction domain located within PML moiety was required for PAX5-PML homodimer complex formation and partial transcriptional repression of genes controlled by PAX5. Fluorescent microscopic examination of transiently expressed YFP-tagged proteins in HeLa and 293T cells demonstrated that YFP-PAX5-PML and YFP-PAX5-PML(ΔCC) exhibited a diffuse granular pattern within the nucleus, similar to PAX5 but PML. By fluorescent recovery after photobleach (FRAP), we have shown that PAX5-PML fusion protein has reduced intranuclear mobility compared to wild-type PAX5. Furthermore, the dimerization domain (coiled-coil) of PML was responsible for the reduced intranuclear mobility of PAX5-PML. These results indicate that the coiled-coil domain of PAX5-PML is important for each of the known activities of PAX5-PML fusion proteins. Disclosures: No relevant conflicts of interest to declare.