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To evaluate the value of circulating cell-free DNA (CFDNA) quantification for screening lymphoma, to analyse the relationship of circulating CFDNA with curative effect under standard therapeutic schedule, and to determine whether circulating CFDNA could be applied to monitor and prognosticate lymphoma.The peripheral blood samples from 32 patients(21 cases of lymphoma and 11 cases of lymphadenitis) with superficial lymph node enlargement were collected, 9 healthy volunteers were as the normal control. Fluorescent quantitative PCR was used to detect the circulating CFDNA in 3 groups. Then, the relationship of circulating CFDNA with common characteristics of lymphoma was analysed, so as to evaluate the importance of circulating CFDNA to the curative effect and prognosis.The circulating CFDNA level in patients with lymphoma was higher than that in patients with lymphadenitis and healthy volunteers (56.71±50.61) ng/ml vs (19.21±15.52) ng/ml and (8.26±7.06) ng/ml (P0.05), but the difference between the latter 2 was not statistically significant (P=0.118). The circulating CFDNA level in lymphoma significantly correlated with the level of lactate dehydrogenase(LDH) (P0.05). ROC analyses revealed that the detection of plasma DNA could discriminate the lymphoma from normal controls with 75% sensitivity, 85% specificity and with a cut-off value of 24.67 ng/ml. The higher circulating CFDNA clearance rate after standard therapy, the higher the rate of complete remission(CR) (P0.05) and the longer overall survival(P0.001).Elevated circulating cell-free DNA levels may be useful as a screening tool for lymphoma. Circulating CFDNA level may serve as a potential indicator for evaluation of the curative effect and prognosis.

Study question What is the function of Spindlin 1 (Spin1) in metaphase II stage oocytes in pigs? Summary answer Depletion of Spin1 induces spontaneous oocyte activation and overexpression of Spin1 causes multinuclear formation through induction of DNA damage in porcine oocytes. What is known already Little is known about the function of Spin1 in oocytes and embryos. In mouse oocytes, Spin1 is specifically expressed during gametogenesis and is essential for meiotic resumption. In somatic cells, Spin1 promotes cancer cell proliferation and activates WNT/T-cell factor signaling. Study design size, duration After knockdown (KD) or overexpression of Spin1 in porcine MII-stage oocytes, MII maintenance was checked following additional culture for 24 h. Investigated parthenotes were cultured up to the four cell (72 h) or blastocyst (7 days) stages. Participants/materials, setting, methods Spin1 was knocked down in porcine oocytes and embryos via microinjection of pig Spin1-targeting siRNA. For Spin1 overexpression, porcine Spin1-eGFP cRNA was generated. Additionally, for rescue experiments, cRNA encoding siRNA-resistant mouse Spin1 was added to the pig Spin1-targeting siRNA. For the overexpression and rescue experiments, microinjection and culture were performed using the same methods as the KD experiments. Main results and the role of chance KD of Spin1 in MII-stage porcine oocytes reduced metaphase-promoting factor and mitogen-activated protein kinase activities, resulting in spontaneous pronuclear formation without calcium activation. However, the DNA damage response was triggered by Spin1 overexpression, generating the checkpoint protein γH2A.X. Furthermore, Spin1 overexpression blocked metaphase-anaphase transition and led to multinucleation in oocytes and embryos. Large scale data None. Limitations, reasons for caution This study is based on in vitro investigations with abnormal expression levels of Spin1. This may or may not accurately reflect the situation in vivo. Wider implications of the findings Spin1 is essential to maintain MII arrest, but a high level of Spin1 induces DNA damage in oocytes and embryos. Thus, a system to accurately regulate Spin1 expression operates in porcine MII-stage oocytes and embryos. Study funding and competing interest(s) This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (No. 2015R1D1A1A01057629). The authors declare no competing financial interests.

Background The unfolded protein response (UPR) is activated following an endoplasmic reticulum (ER) stress. The aim of this study was to investigate the global expression of UPR genes in human OA chondrocytes in induced (I)-UPR conditions, and to explore the regulation and role of the UPR genes in homeostatic (H)-UPR conditions in human normal and OA chondrocytes. Methods Gene expression was determined by PCR array and qPCR. Protein production in cartilage was determined by immunohistochemistry, gene silencing by specific siRNAs, and gene regulation by treating chondrocytes with cytokines and growth factors associated with cartilage pathobiology. Results Several UPR genes, among them ERN1, PERK, and CREB3L2 were downregulated in OA compared to normal chondrocytes at both the mRNA and protein levels, but the ER stress response triggered by thapsigargin or tunicamycin treatment was similar in normal and OA chondrocytes. The activation of ER stress sensors (phosphorylated PERK, cleavage of ATF6B, and the spliced mRNA forms of XBP1) was not significantly increased in OA chondrocytes/cartilage. PDGF-BB and IL-6 significantly downregulated the expression of ERN1, PERK, and CREB3L2, but not that of ATF6B. Silencing experiments done under conditions of no ER stress (physiological conditions) revealed that decreasing ERN1 expression led to decreased COL2a1, MMP-13, ADAMTS4 and ADAMTS5 expression, while decreasing CREB3L2 and ATF6B led to decreased ADAMTS5 and ADAMTS4 expression, respectively. Importantly, the downregulation of PERK expression increased COL1a1 and suppressed COL2a1 expression. Conclusions Although the level of ER stress is not significantly increased in OA chondrocytes, these cells respond strongly to an acute ER stress despite the decreased expression of ERN1, PERK, and CREB3L2. Emerging findings revealed for the first time that these genes play a role in cartilage biology in conditions where an acute ER stress response is not triggered and OA is not characterized by an overall basal activation of the ER stress response. Importantly, these findings identify PERK as a potential target for new OA treatment avenues. Electronic supplementary material The online version of this article (doi:10.1186/s13075-016-1070-6) contains supplementary material, which is available to authorized users.

Purpose To assess the benefits of using a low-frequency convex probe and detecting ophthalmic artery blood flow for the differential diagnosis of occlusion from severe stenosis of the internal carotid artery (ICA). Methods A total of 103 patients were enrolled and had three sequential examinations by an experienced sonographer. First, the proximal ICA was examined with a 5.0–7.5-MHz linear probe. Second, the distal ICA was examined with a 3.5–5.0-MHz convex probe. Third, the ophthalmic artery (OA) was examined with a 5.0–7.5-MHz linear probe. Three parallel sets of diagnoses were made based on results from (1) high-frequency ultrasonography; (2) combined use of high- and low-frequency ultrasonography; and (3) combined use of high- and low-frequency ultrasonography along with OAs examination findings. Efficiencies of the three diagnoses were compared by receiver operating characteristic curves with digital subtraction angiography as the gold standard. Results The second diagnostic approach yielded the highest sensitivity (96.6%) and specificity (94.5%). OA blood flow was reversed in most, but not all, cases of ICA occlusion (27/29). Areas under the receiver operating characteristic curves of the three diagnoses were significantly different (p < 0.05). Conclusions The diagnostic performance of combined high- and low-frequency ultrasonography is greater than that of high-frequency ultrasonography alone. Although OA examination does not increase the diagnostic performance, it provides helpful data for the assessment of hemodynamics and collateral circulation. © 2013 Wiley Periodicals, Inc. J Clin Ultrasound 41:408–414, 2013

The city is a high-density human settlement. The contradiction in the city between human activity and ecological environment is particularly serious. In order to introduce eco-civilization into the city construction, it is inevitable to construct eco-civilization city. This article analyzes the connotation and characteristics of eco-civilization city, the main theoretical framework of urban eco-civilization construction including eco-civilization city construction planning, environmental protection, ecological economy/industry, ecological security, ecological concept/act, ecological culture, ecological politics/government, ecological law/system/policy, and ecological and livable city, which is expected to provide a reference for realizing the coordinated development of regional economic and environment.

The construction of eco-city is meant to harmonize the social, economical and natural development, with high-efficiency use of the material, energy and information. Eco-city is the inevitable choice of a city development in future. It is very important to explore the eco-city construction in the context of ecological civilization. In the background of ecological civilization, the relationship between ecological civilization and eco-city construction is explored. Furthermore, the importance of eco-city construction has been discussed. Finally, the eco-city theories and practical experiences abroad are summarized and some opinions and advices on eco-city construction in Shenyang, China are given.

From October 2009 to September 2010, this study focused on investigation the performance of a subsurface wastewater infiltration (SWI) system in treating domestic sewage, and evaluated the potential of the surface water pollution by using the SWI system. The results showed the removal efficiencies were relatively high: for biochemical oxygen demand (BOD), chemical oxygen demand (COD), suspended solids (SS), ammonia nitrogen (NH3-N) and total phosphorus (TP) were 95.0, 89.1, 98.1 87.6 and 98.4%, respectively. Meanwhile, the monitoring of a nearby stream characteristic indicated the SWI system could not decrease the receiving surface water quality.

To investigate whether Gli1 expression is important in relapse after radical operation of breast cancer.Using immunohistochemistry, Gli1 expression was analyzed in human primary breast cancer (n=284) and paracancerous tissues (n=20), and also in local lymph nodes (n=28) and metastatic lymph nodes (n=28).Initial analysis of Gli1 expression in a small cohort of 20 breast tumors and their paracancerous tissues showed a tendency towards Gli1 overexpression in breast cancer tissues (P0.001). Further, Gli1 expression in 284 breast cancer tissue samples was analyzed and a significant correlation was found between increased expression of nuclear Gli1 and unfavorable recurrence-free survival (RFS) (P0.05). The nuclear expression of Gli1 in metastatic lymph nodes following relapse after radical operation was much higher than that in the local lymph nodes of primary carcinoma (P0.05). Most interestingly, the expression of Gli1 was much higher in the interstitial tissues of the relapsed group than of the non-relapsed group (P0.001).Breast cancer shows a high prevalence of Gli1 expression, which is significantly correlated with aggressive features and unfavorable RFS. Nuclear Gli1 overexpression, especially in the interstitial tissues, signified early relapse after radical operation of breast cancer.

Radial oxygen loss (ROL) from roots of reed, typha and wild rice stem was investigated. Root excretion oxygen showed that the rates of the ROL increased with the illumination time under hard light intensity. For each plant, the ability that roots excreting oxygen under different light intensities decreased successively in the order of hard light, non-light and weak light. The ROL was different with the increased sequence of typha, wild rice stem and reed. The local available reed and typha could remove the pollutants effectively in the pilot test with the average removal rates of 41.99%, 28.47%, 48.38% and 48.76%, respectively.

Subsurface wastewater infiltration treatment system (SWIS) is a domestic wastewater treatment technology. While the traditional technology has a large floor area, long starting period and low pollutants removal rate. A new biosubstrate that can be applied in SWIS is studied, experiments about its characteristics were performed, and treatment effect was compared. The results show that the biosubstrate has high biological activity, with it, SWIS has shorter starting period, better pollutants removal rate and consequently needs less floor space.

A novel ion-imprinted organic-inorganic hybrid sorbent was synthesized and characterized by FT-IR and nitrogen gas adsorption-desorption. The adsorption property and selective recognition ability of the imprinted sorbents for the lead ion were studied. Results showed that the uptake capacities and selectivity coefficients of the double imprinted sorbent were much higher than that of the non-imprinted sorbent. The adsorption capacity of the double imprinted sorbent is 545.6 mg•g-1. The largest relative selectivity coefficient between Pb (II) and Cd (II) was 192. This results suggested that the new sorbent can be used as effective solid-phase material for the selective preconcentration and separation of Pb (II) in environmental water samples.

Wastewater composition and characteristic in a northern university of China were analyzed according to the sampling and analysis of sewage discharged from the different function units. The sewage discharge quantities and principles of different time points and seasons were monitored. The results showed that the concentrations of pollutants in northern campus sewage changed in a certain pattern with the alteration of sampling time and season. Different function units had significant effect on wastewater composition. The influence degree depends on the concentration of characteristic contaminant. The study will helpful to design different campus sewage treatment system due to the principle of partition and dispersion.

Aiming at the ultrasonic vibration polishing technology of silicon wafer, this paper proposes a disc-shaped ultrasonic piezoelectric polishing vibrator of out-of-plant bending travalling wave. Basing on numerical analysis, the principle prototype of the polishing vibrator has been designed and produced, and then tested. This polishing vibrator can be used in polishing process of semiconductor silicon wafer.

Mitochondria are important regulators of both apoptosis and autophagy. One of the triggers for mitochondrial-mediated apoptosis is the production of reactive oxygen species (ROS), which include hydrogen peroxide, superoxide, hydroxyl radical, nitric oxide and peroxynitrite. Recently, several studies have indicated that ROS may also be involved in the induction of autophagy. In the present study, we used H(2)O(2) to induce mitochondrial stress, examined apoptotic- and autophagic-related gene expression and observed LC3 protein (autophagosome presence marker) expression in porcine parthenotes developing in vitro. In porcine four-cell parthenotes cultured for 5 days in NCSU37 medium containing 0.4% BSA, the developmental rate and mitochondrial distribution did not differ from that of the group supplemented with 100 µM H(2)O(2) but was significantly decreased in the group supplemented with 500 µM H(2)O(2) (P

Hydroxyapatite/Chitosan (HAp/CS) bio-coatings were prepared on the surface of carbon/ carbon (C/C) composites by hydrothermal electrophoretic deposition, using sonochemical process resulted HAp nanoparticles, isopropyl alcohol and chitosan as raw materials. The influences of hydro- thermal conditions and deposition voltage on the microstructures and morphologies of the as-prepared coatings were investigated. It was shown that homogenous and dense HAp/CS coatings on C/C composites are obtained by hydrothermal electrophoretic deposition. With the increase of deposition voltage, density and homogeneity of the as-prepared HAp/CS composite coatings are well improved. Due to the growth of HAp nanoparticles in the hydrothermal condition, the subsequent heat treatment of the HAp/CS coatings is not needed.

Seven three-dimensional (3D) cadmium coordination frameworks, {[Cd(5MT)2] (1a); [Cd5(N3)(5MT)9·0.12H2O] (1b); [Cd3(OH)Cl1.39(N3)0.61(5MT)3] (1c), 5MT = 5-methyl-tetrazole}; {[Cd3(OH)Cl(N3)(5AT)3 (2a); Cd2(OH)Br(5AT)2 (2b), 5AT = 5-amino-tetrazole]}; {[Cd7Cl2(5BT)12(H2O)2 (3a); and [Cd7Br2(5BT)12(H2O)2 (3b), 5BT = 5-butyl-tetrazole], were obtained through in situ tetrazole synthesis and have been structurally characterized by elemental analyses, Fourier transform infrared spectroscopy, thermal studies, and single crystal and powder X-ray diffraction. Compounds 1a and 1b were constructed by the interconnection of 5MT and cadmium metal ions, and both exhibited distinct topological nets: 1a has a diamond-like framework, while 1b has the acs topological net. Compounds 1c and 2a represented similar structural network motifs; both were intricate coordination polymers constructed from interconnected {ClCd3O} subnets through tetrazole anions. Compound 2b was constructed from two-dimensional (2D) wavelike {BrCd3O} ...

Clinically diagnosed cases of Babesia gibsoni infection in dogs were evaluated on the basis of polymerase chain reaction (PCR) results and nucleotide sequences of the 18S rRNA gene. Twenty-nine of 117 submitted dogs were PCR positive. The breed of dogs infected with B. gibsoni varied; however, pit bull terriers were the breed most often infected. The infection rate was higher in densely populated regions and in male dogs. Young dogs (age < 3 years) were more sensitive to B. gibsoni infection, and 6 out of the 29 infected dogs had a history of tick exposure. The clinical signs observed during physical examination were related to canine babesiosis, and many dogs showed symptoms similar to those associated with anemia. The results of hematologic analysis revealed severe hemolytic anemia and thrombocytopenia in the infected dogs. However, the blood smears of 29 infected dogs showed very low levels of parasitemia. Nucleotide sequencing of the PCR products revealed that the 18S rRNA gene sequences of B. gibsoni in South Korea were very close to those reported in Spain, Japan, and the United States.

Kinetic study on the complexation of gallic acid with ferrous sulfate was performed using UV-Vis absorption spectroscopy. Under the experimental conditions, the stoichiometric composition of the formed complex is 1:1. The complexation reaction was found to be a second-order one. The influences of temperature, ionic strength and solvents on the complexation reaction were investigated. According to the Arrhenius equation, the apparent activation energy of the complexation reaction was evaluated to be 71.64 kJ x mol(-1). A three-step reaction mechanism was proposed, which can well explain the kinetic results obtained.

Biomass continues to be an important candidate as a renewable resource for energy, chemicals, and feedstock. Decomposition of biomass in high temperature liquid water is a promising technique for producing industrially important chemicals such as 5-hydroxymethylfurfural (5-HMF), furfural, levulinic acid with high efficiency. Hexose, which is the hydrolysis product of cellulose, will be one of the most important starting chemicals in the coming society that is highly dependent on biomass. Taking fructose as a model compound, its decomposition kinetics in organic acids-enriched high temperature liquid water was studied in the temperature range from 180 °C to 220 °C under the pressure of 10 MPa to further improve reaction rate and selectivity of the decomposition reactions. The results showed that the reaction rate is greatly enhanced with the addition of organic acids, especially formic acid. The effects of temperature, residence time, organic acids and their concentrations on the conversion of fructose and yield of 5-HMF were investigated. The evaluated apparent activation energies of fructose decomposition are 126.8 ± 3.3 kJ mol−1 without any catalyst, 112.0 ± 13.7 kJ mol−1 catalyzed with formic acid, and 125.6 ± 3.8 kJ mol−1 catalyzed with acetic acid, respectively, which shows no significant difference.

Six lanthanide coordination polymers La2(1,4-BDOA)2(OX)(H2O) (1), Ln2(1,4-BDOA)2(OX)(H2O) [Ln = La (2); Ln = Pr (3); Ln = Sm (4); Ln = Eu (5); Ln = Tb (6)], [BDOA = benzene-1,4-dioxydiacetate; OX = oxalate] had been prepared under hydrothermal conditions, which were characterized by elemental analysis, infrared spectroscopy, thermogravimetric analysis, and single-crystal X-ray diffraction. Compounds 1–6 crystallize in a triclinic system, space group P 1 ¯ ; compounds 1 and 2 are true structural supramolecular isomers based on BDOA ligands constructed from different acid effects; compounds 2–5 are isostructural and exhibit new 3D 12-connected topological networks, which are the same as the topological structure of 6, while 1 shows another 3D 11-connected topological architecture. Furthermore, the luminescent properties of 5 and 6 were studied.

By control of mixed organic ligands with different geometries, three unusual 4d–4f heterometallic coordination polymers, [AgLn(nic)2(ox)] · 2H2O [Ln = Eu (1); Dy (2); Ho (3)] [nic = nicotinate; ox = oxalate] have been synthesized under hydrothermal reaction. Three structures represent 3D open heterometallic coordination frameworks that are constructed from rare zigzag lanthanide-ox-silver chains and nic linkers. Furthermore, the luminescent property of complex 1 was discussed.

Four transition–lanthanide metal–organic coordination polymers, namely [Ag2Ln(nic)4(H2O)4 · (ClO4) · H2O] [Ln = Eu (1), Gd (2)] and [AgLn(nic)2(ox)0.5(H2O)2 · (ClO4) · H2O] [Ln = Tb (3), Yb (4)] (nic = nicotinate; ox = oxalate) have been synthesized by the hydrothermal reactions of 4d and 4f metal salts with N-/O-donor ligands. The isostructural complexes 1 and 2 exhibit novel 2D wave-like heterometallic layers constructed by the assembly of 1D chains of lanthanide–carboxylate with Ag(nic)2 subunits. Complexes 3 and 4 show another unusual 3D heterometallic coordination framework constructed from 2D lanthanide–oxalate layers and pillar-like Ag(nic)2 subunits. Furthermore, the luminescent properties of complexes 1 and 3 were studied.

The development of polarized hippocampal neurons with a single axon and multiple dendrites depends on the activity of phosphoinositide 3-kinase (PI3K) and the GTPase Rap1B. Here we show that PI3K regulates axon specification and elongation through the GTPase Rheb and its target mammalian target of rapamycin (mTOR). Overexpression of Rheb induces the formation of multiple axons, whereas its suppression by RNA interference blocks axon specification. mTOR is a central regulator of translation that phosphorylates eIF4E-binding proteins like 4E-BP1. Axon formation was suppressed by inhibition of mTOR and expression of mTOR-insensitive 4E-BP1 mutants. Inhibition of PI3K or mTOR reduced the level of Rap1B, which acts downstream of Rheb and mTOR. The ubiquitin E3 ligase Smurf2 mediates the restriction of Rap1B by initiating its degradation. Suppression of Smruf2 by RNA interference is able to compensate the loss of Rheb. These results indicate that the mTOR pathway is required to counteract the Smurf2-initiated degradation of Rap1B during the establishment of neuronal polarity.

The self-assembly of pyrazine-2-carboxylate and oxalate with mixed-metal salts under hydrothermal conditions gave three isostructural 3D 4d-4f coordination polymers, LnAg(ox)(2-pzc)2 · H2O [Ln = Eu (1); Tb (2); Dy (3)] [ox = oxalate, 2-pzc = pyrazine-2-carboxylate]. All three structures exhibit same unusual 3D nanoporous heterometallic coordination frameworks constructed by zigzag lanthanide–oxalate chains and Ag(2-pzc)2 units. Furthermore, the luminescent properties of complexes 1 and 2 are discussed.

A 3d–4f heterometallic coordination polymer, {[Cu10Er4(IN)14ClBr7(H2O)10] · 2H2O}n (1) [IN = isonicotinate], has been synthesized by hydrothermal reaction of the ligand with CuBr2, NaCl and Er2O3. Complex 1 represents a rare 3D heterometallic coordination framework constructed from Er-carboxylate subunits and unique Cu-halide (Br/Cl) chains that are bridged by a single type of linear ligand. Furthermore, the luminescent properties of complex 1 have also been investigated.

Ehrlichia chaffeensis is one of the causative agents of canine ehrlichiosis and human monocytic ehrlichiosis (HME). Canine ehrlichiosis caused by E. chaffeensis was diagnosed in two dogs in South Korea based on clinical findings, and the diagnosis was confirmed by polymerase chain reaction (PCR) and DNA sequencing. A 5-year-old intact male American Pit bull terrier allowed outdoors was found to be concurrently infected with Babesia gibsoni and E. chaffeensis. The major clinical findings were lethargy and reddish urine, and laboratory analysis revealed severe hematuria and thrombocytopenia. In addition, a 3-year-old neutered male Shih-tzu was also found to be infected with E. chaffeensis. Although this dog was an indoor companion animal, he was frequently allowed outside for exercise. The clinical signs observed in this dog included generalized purpura with petechiae and ecchymoses due to thrombocytopenia. A 390-bp partial portion of E. chaffeensis 16S rRNA gene was amplified in both cases, and nucleotide sequence analysis revealed 99% homology of this fragment with other E. chaffeensis isolates. These findings demonstrate the presence of E. chaffeensis infection in dogs in South Korea, and this is the first report to confirm clinical cases of E. chaffeensis infection in dogs.

Two unusual d10 compounds, [Zn2(bipy)3(H2O)8(ClO4)2(paba)2].2(bipy).4H2O (1) and [Cd2(bipy)4(H2O)6(ClO4)2(paba)2].(bipy).5H2O (2) (bipy = 4,4'-bipyridine, paba = p-aminobenzoate), were obtained from reaction of the metal salts, bipy and paba in an EtOH/H2O mixture. The bipy ligands in the two compounds exhibit two new modes of coordinating to transition metal ions, resulting in the formation of large porous frameworks. Immersion of single crystals of 1 in an aqueous solution of NH4PF6 results in the formation of its hexafluorophosphate derivative 3 as shown by single crystal diffraction. Immersion of crystals of 3 in NaClO4 regenerates 1. Furthermore, compound 1 also shows interesting anion sensing properties in an EtOH/H2O mixture.

The projector augmented wave method (PAW) within the Perdew–Burke–Ernzerhof (PBE) form of generalized gradient approximation (GGA) is applied to investigate the effect of hydrostatic pressure on the structures of titanium metal. In our calculations, the full geometry optimization for all the structures at each volume considered is performed. The phase sequence ω → γ → δ has been obtained and is in good agreement with the experimental results of Vohra et al. and Akahama et al. The obtained phase transition δ → bcc is at ca. 160.9 GPa. This conclusion is in accordance with those of Kutepov et al. and Joshi et al. The quasi-harmonic Debye model for the first time is used to predict phase diagram of α → ω and the result is in better agreement with the experimental data than those from earlier calculations at 300 K.

To explore the effect of overexpression of SH2-containing tyrosine phosphatase 1 (SHP-1) on sensitivity of chronic myelogenous 1eukemia (CML) K562 cell line to imatinib and its related mechamism.K562 cells were infected with the lentiviral plasmids containing the specified retroviral vector (pEX-SHP-1-puro-Lv105) or the mock vector (pEX-EGFP-puro-Lv105). The expression of SHP-1 in K562 cells treated with 0.2 µmol/L imatinib (IM) for 72 h was determined by Western blot. After transfection the CCK-8 assay was used to determine the proliferation of the tramfected K562 cells (K562(SHP-1) and K562(EGFP) cells) at 72 h after exposure to different doses of IM, the half inhibitary concentration (IC50) was calculated. The mechanisms of the overexpression effects of SHP-1 and IM on the proliferation in K562 cells was investigated, the BCR-ABL1 activity and the level of tyrosine phosphorylation of CrkL (pCrkL) was measured by flow cytometry; the Western blot was used to detect the expression and activity of these molecules controlling cell growth, including MAPK, AKT, STAT5 and JAK2.After exposure of K562 cells to 0.08 µmol/L IM for 72 h, there was no significant change of SHP-1 expression in K562 cells. After exposure to 0.2 µmol/L of IM for 72 h, the inhibitory rate of K562(SHP-1) group was higher than that of K562(EGFP) group (P0.05), indicating that overexpression of SHP-1 in K562 cells could enhance the proliferation inhtibition effect of IM on K562 cells. The IC50 of IM in K562(SHP-1) cells was the lower as compared with that of K562(EGFP) cells (P0.05) after exposure to different concentrations of IM for 72 h. The slope of K562(SHP-1) cells was the largest ranging 0.02 - 0.16 µmol/L of IM. Overexpression of SHP-1 and IM could inhibit the activity BCR-ABL1, MAPK, AKT, STAT5 and JAK2 signaling pathways in the K562 cell line and displayed a synergistic effect.SHP-1 inhibits BCR-ABL1, MAPK, AKT, STAT5 and JAK2 signaling pathways in K562 cells, the overexpression of SHP-1 can enhance the sensitivity of K562 cells to IM.

Chronic myeloid leukemia is a myeloproliferative disorder characterized by excessive cloning of bone marrow multipotent stem cells. According to the disease course, the CML may be divided into chronic phase (CP), accelerated phase (AP) and blastic phase (BP). At present, the molecular mechanisms of acute transformation of CML has not been fully understood. The recent studies have shown that the epigenetics is one of mechanisms in blastic transformation of CML, including three molecular mechanisms such as DNA modification, histone modifications and RNA-related dysregulation. The molecular mechanisms for epigenetics leading to the transformation of CML are discussed in this review.

Identification of bioactive compounds directly from complex herbal extracts is a key issue in the study of Chinese herbs. The present study describes the establishment and application of a sensitive, efficient, and convenient method based on surface plasmon resonance (SPR) biosensors for screening active ingredients targeting tumor necrosis factor receptor type 1 (TNF-R1) from Chinese herbs. Concentration-adjusted herbal extracts were subjected to SPR binding assay, and a remarkable response signal was observed in Rheum officinale extract. Then, the TNF-R1-bound ingredients were recovered, enriched, and analyzed by UPLC-QTOF/MS. As a result, physcion-8-O-β-D-monoglucoside (PMG) was identified as a bioactive compound, and the affinity constant of PMG to TNF-R1 was determined by SPR affinity analysis (K D = 376 nM). Pharmacological assays revealed that PMG inhibited TNF-α-induced cytotoxicity and apoptosis in L929 cells via TNF-R1. Although PMG was a trace component in the chemical constituents of the R. officinale extract, it had considerable anti-inflammatory activities. It was found for the first time that PMG was a ligand for TNF receptor from herbal medicines. The proposed SPR-based screening method may prove to be an effective solution to analyzing bioactive components of Chinese herbs and other complex drug systems. Graphical abstract Scheme of the method based on SPR biosensor for screening and recovering active ingredients from complex herbal extracts and UPLC-MS for identifying them. Scheme of the method based on SPR biosensor for screening and recovering active ingredients from complex herbal extracts and UPLC-MS for identifying them.

In order to investigate the influencing factors of 5-hydroxymethyl-2-furaldehyde (5-HMF) content in Schisandra, confirm the theory of 5-HMF deriving mainly from Schisandra processing course, and give some suggestions about the Schisandra processing method, the 5-HMF contents in decoctions of Schisandra under different heating temperature, decocting time, soaking time, processing methods and treatment with different solvents before decocting the Schisandra were measured by RP-HPLC method. The results showed that there is great difference of 5-HMF level in decoctions from differently processed Schisandra and unprocessed Schisandra; decocting time of 60 min has some effects on 5-HMF level in decoctions and there is certain quantity 5-HMF in processed Schisandra itself and very little 5-HMF in unprocessed Schisandra. Heating time, heating temperature and treating solvents all have effect on 5-HMF level in decoction of Schisandra. 5-HMF in Schisandra was mainly from processing course. Both long heating time and high heating temperature can increase 5-HMF level in Schisandra. The production of 5-HMF in Schisandra may have some relationships with some polar components, which can dissolve in water, ethanol and acetone, especially in ethanol. To control processing temperature, processing time and treatment with some solvent is very important for controlling 5-HMF level in Schisandra.

To investigate the expression and clinical significance of DNA methyltransferases (DNMT) mRNA in patients with chronic myeloid leukemia (CML).The expression levels of DNMT mRNA in mononucllear cells (MNC) of bone marrow or in peripheral blood of 93 CML patients in 3 different phases and 10 normal controls (NC) were detected by SYBR Green flurescent quatitative PCR.The relative expression levels of DNMT1 mRNA in NC, chronic phase CML (CML-CP), accelerated phase (CML-AP) and blastic phase (CML-BP) were 1.45 ± 0.22, 1.83 ± 0.63, 2.95 ± 0.87 and 3.24 ± 1.39 resectively. The expression of DNMT1 mRNA showed no statistically significant difference between CML-CP and NC (P = 0.28). The expression of DNMT1 mRNA in advanced stages (including CML-AP and CML-BP) of CML obviously increased in comparison with CML-CP and NC (P0.05). The expression of DNMT1 mRNA in CML-AP was not significantly different from that in CML-BP (P = 0.336). The relative expression levels of DNMT3a mRNA in NC, CML-CP, CML-AP and CML-BP groups were 1.29 ± 0.34, 1.34 ± 0.46, 2.33 ± 1.05 and 3.18 ± 1.23 resectively. And the expression levels of DNMT3a mRNA were not statistically significantly different between CML-CP and NC (P = 0.844). The results showed that the expression of DNMT3a mRNA in the advanced phase of CML significantly increased in comparison with that in CML-CP and NC (P0.05). Meanwhile, the expression of DNMT3a mRNA in CML-AP was not different from that in CML-BP (P = 0.304). The relative expression levels of DNMT3b mRNA in NC, CML-CP, CML-AP and CML-BP groups were 1.37 ± 0.31, 16.41 ± 22.50, 9.36 ± 5.50 and 12.17 ± 13.44 resectively. It was also found that the level of DNMT3b mRNA in CML significantly increased in comparison with NC (P0.05), and that the between the 3 different phase of CML was not statistically significantly different (P0.05).The expression of DNMT mRNA increases in advanced CML as compared with normal controls and CML-CP, and the increased levels of DNMT mRNA probably correlate with disease progression in CML.

Chronic myeloid leukemia (CML) is a myeloproliferative disorder, characterized by excessive proliferation of myeloid cells. CML patients in early phase [also known as chronic phase (CP)] usually respond to treatment with tyrosine kinase inhibitors (TKI), some patients respond initially to TKI, but later become resistant, then resulting in the transformation from CP to more advanced phase, which were subclassified as either accelerated phase or blastic phase. At present, the molecular mechanisms of CML have been not yet clear, and acute transformation has been not fully understood, studies have shown that genomic instability promotes the acute conversion of CML. This review discusses the molecular mechanisms leading to the transformation of CML, and some therapeutic approaches.

Salmonella serovar Pullorum is a causative agent of pullorum disease (PD) in poultry and is responsible for severe economic losses to the poultry industry in many parts of the world. A definitive detection of Pullorum requires culture followed by serotyping and biochemical identification, a process that is tedious and takes several weeks to accomplish. We have developed a rapid allele-specific polymerase chain reaction (PCR) method based on the nucleotide polymorphism in rfbS gene sequence for the serotype-specific detection of Pullorum and its differentiation from the closely related Gallinarum. The specificity of this PCR assay was tested using DNA samples from Pullorum (n = 13), Salmonella serotypes other than Pullorum (n = 19), and closely related non-Salmonella organisms (n = 5). The PCR assay was highly serotype-specific as the PCR amplicon of 147 base pairs was observed only in the case of Pullorum, while all the other DNA samples tested PCR negative. A definitive identification of Pullorum cultures was possible in less than 3 hr. As little as 100 pg of SP DNA was detected. This allele-specific PCR method is highly specific as well as sensitive and may be an effective molecular tool in the rapid and serotype-specific detection of Pullorum and differentiation from other Salmonella species.

Dynamic compression experiments of bismuth (Bi) in a shock pressure range of 38-100 GPa were conducted using an explosive drive. The time-resolved particle velocity profiles at the Bi/LiF (lithium fluoride) interface and the impact velocities were recorded with a displacement interferometer system for any reflector (DISAR). Shock velocities of Bi were measured based on the shorted-pin technique. Sound velocities of Bi were obtained from the peak state time duration measurements with the overtake method. Uncertainties in the measured Hugoniots and sound velocities were analyzed carefully. The new Hugoniot relation, Us = (1.666 ± 0.057) km/s + (1.596 ± 0.037)Up (1.13 km/s ≤Up≤2.05 km/s), was determined by fitting the measured shock velocity and particle velocity to a line. The relation between the Grüneisen coefficient and the density, γρ=const., is found to be a good approximation to describe the sound velocities of Bi above the shock-induced melting pressure, by carefully analysis of the sound velocity data.

Chronic lung disease (CLD) of prematurity remains a significant cause of morbidity among premature infants. It is a multifactorial disorder and characterized by an early increased number of neutrophils and alveolar macrophages, with later architectural epithelial and endothelial cell damage. Recently, apoptosis of type 2 pneumocytes in the lung of preterm neonates with acute and chronic lung disease has been examined and apoptosis of mesenchymal cells was detected in the chronic stage of bronchopulmonary dysplasia. Infection and inflammatory responses in the lungs play important roles. However, the contribution of Ureaplasma urealyticum to the development of CLD is debated. We found that U. urealyticum induced apoptosis in human type II lung epithelial cells (A549 cell line) and macrophages (derived from human monocytic cell line THP-1) by measuring the outer leaflets translocation of phosphatidylserine (flow cytometry analysis and fluorescence microscopy assessment), DNA fragmentation analysis, cell morphology changes such as diminution in cell volume, increased cytoplasmic staining, and nuclear pyknosis (hematoxylin and eosin staining) and viable counting (trypan blue exclusion). Anti-TNF-α monoclonal antibody partially protected the macrophages from undergoing apoptosis after infection with U. urealyticum. Our findings imply that U. urealyticum might be involved in impairing lung structure and host immune response during the development of CLD.

This study was aimed to investigate the expression level of SHP-1 and C-kit genes in acute leukemia HL-60 cells and effect of 5-aza-CdR demethylation on expression of SHP-1 and C-kit genes. RT-PCR was used to detect the mRNA expression level of SHP-1 and C-kit mRNA in HL-60 cells of the drug-treated group and control group.The methylation specific PCR (MSP) was applied to measure the methylation status of SHP-1 and C-kit genes in HL-60 cells.The results showed that after being treated with 5-aza-CdR, the recovery of SHP-1 gene expression was observed in HL-60 cells in which SHP-1 mRNA originally was not expressed. Meanwhile, the high expression level of C-kit mRNA in HL-60 cells was decreased. When HL-60 cells were treated with 0, 0.5, 1.0, 2.0 µmol/L 5-aza-CdR, the demethylation effect was enhanced, the expression of SHP-1 mRNA displayed an ascending tendency, and the expression of C-kit mRNA showed an descending tendency in dose-dependent manner (P0.05) . It is concluded that the absence of SHP-1 mRNA expression in HL-60 cells and recovery of expression after treatment with 5-aza-CdR suggest that the hypermethylation of SHP-1 gene relates with pathogenesis of leukemia, and the abnormal increase of C-kit mRNA expression maybe exist in formation of leukemia. The effect of 5-aza-CdR on expression of SHP-1 and C-kit shows dose-dependency, the higher the 5-aza-CdR concentration, the higher the SHP-1 expression and the lower the C-kit expression, moreover, the effect of 5-aza-CdR shows time-dependency in specific concentration.The SHP-1 mRNA expression negatively correlates with C-kit mRNA expression, suggesting that the decrease or absence of SHP-1 expression in leukemia cells weakens the negative regulation on C-kit signaling pathway, thus plays a role in the formation of leukemia.

The study aims to investigate the relationship between nuclear factor (nuclear factor kappa B (NF-κB)) viability and lactacystin-mediated cell apoptosis in gastric cancer cells. Two gastric cancer cell lines (MKN28 and SGC7901) were treated with lactacystin-a proteasome inhibitor for 24 h. The cell viability, toxicity, and death were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. DNA binding viability of NF-κB and caspase-3 viability were analyzed by ELISA; the expression of p65 NF-κB nuclear protein was detected by immunocytochemistry and Western blot. Lactacystin reduced DNA binding viability of NF-κB (t = 3.0,P = 0.013) and the NF-κB viability (compared to the 5, 10 μmol/L MKN28 cell (p53 mutant) line, P < 0.001) and the expression of p65 NF-κB nuclear protein decreased parallelled to concentrations of lactacystin in MKN28 cell line, while without obvious effects on NF-κB viability in SGC7901 cell line (P = 0.381), while the viability of caspase-3 increased also along with the raising of lactacystin concentrations (compared to control, 5 μmol/L: SGC7901 cell line P = 0.029, MKN28 cell line P < 0.001; 10 μmol/L: SGC7901 cell line, P < 0.001, MKN28 cell line, P < 0.001). It was concluded that lactacystin had diversified killing effects on gastric cancer cells. The mechanism may be related to induce the apoptosis by downregulation of nuclear factor kappa B viability. There may be additional cell survival/death pathway in SGC7901 gastric cancer cells.

Staphylococcus aureus bacteraemia (SAB) originating from local infections can lead to severe secondary infections such as endocarditis. The protective effect of antibodies against secondary infections was studied in a rat model, where a local joint infection leads to bacteraemia and endocarditis on damaged aortic valves. In this study, immunizations with a truncated D2-domain of the S. aureus fibronectin-binding protein displayed on a cow-pea mosaic virus (CPMV-D) carrier induced protection against endocarditis (P < 0.05). Opsonization of S. aureus with antibodies raised against CPMV-D stimulated both neutrophil activity and macrophage phagocytosis in vitro. Furthermore, intravenous administration of these antibodies protected mice from weight loss due to SAB.

Chronic lung disease (CLD) of prematurity is an inflammatory disease with a multifactorial etiology. The importance ofUreaplasma urealyticumin the development of CLD is debated, and steroids produce some improvement in neonates with this disease. In the present study, the capability ofU. urealyticumto stimulate rat alveolar macrophages to produce nitric oxide (NO), express inducible nitric oxide synthase (iNOS), and activate nuclear factor κB (NF-κB) in vitro was characterized. The effect of NO on the growth ofU. urealyticumwas also investigated. In addition, the impact of dexamethasone and budesonide on these processes was examined. We found thatU. urealyticumantigen (≥4 × 107color-changing units/ml) stimulated alveolar macrophages to produce NO in a dose- and time-dependent manner (P< 0.05). This effect was further enhanced by gamma interferon (100 IU/ml;P< 0.05) but was attenuated by budesonide and dexamethasone (10−4to 10−6M) (P< 0.05). The mRNA and protein levels of iNOS were also induced in response toU. urealyticumand inhibited by steroids.U. urealyticumantigen triggered NF-κB activation, a possible mechanism for the induced iNOS expression, which also was inhibited by steroids. NO induced byU. urealyticumcaused a sixfold reduction of its own growth after infection for 10 h. Our findings imply thatU. urealyticummay be an important factor in the development of CLD. The host defense response againstU. urealyticuminfection may also be influenced by NO. The down-regulatory effect of steroids on NF-κB activation, iNOS expression, and NO production might partly explain the beneficial effect of steroids in neonates with CLD.

Curli organelles are expressed by commensal Escherichia coli K12 and by Salmonella typhimurium at temperatures

The self-assembly of nicotinic acid and sodium oxalate with mixed 3d–4f metal salts under hydrothermal conditions gave three isostructural 3D 3d–4f coordination polymers, [LnCu(nic)2(ox)] · xH2O [Ln = La, x = 1 (1); Ln = Eu, x = 2 (2); Ln = Gd, x = 2 (3)] [ox = oxalate, nic = nicotinate]. All three structures exhibit same unusual 3D microporous heterometallic coordination frameworks that are built up by rare tetranuclear Ln2Cu2 clusters and mixed ox and nic linkers. Furthermore, the luminescent property of complex 2 has also been investigated.

Catechol-O-methyltransferase inhibitors have been newly introduced as adjunct drugs to the levodopa/dopa decarboxylase inhibitor therapy in Parkinson's disease. When given alone, catechol-O-methyltransferase inhibitors seem to affect behaviour. We wanted to determine whether the concentrations of free amine would be increased by catechol-O-methyltransferase inhibition with tolcapone and underpin the positive behavioural effects. To this end, dopamine and noradrenaline levels were analyzed in the microdialysis perfusion fluid collected from several brain regions in chloral hydrate anaesthetized rats. We also analyzed the turnover rate of catecholamines in the brain after single doses of tolcapone and entacapone using the alpha-methyl-p-tyrosine method. On their own, tolcapone (at 10 or 30 mg/kg) did not elevate dopamine or noradrenaline levels in any brain region studied although the formation of catechol-O-methyltransferase-dependent metabolites was strongly reduced. Neither tolcapone nor entacapone (at 30 mg/kg) affected the turnover rate of catecholamines. It seems that catechol-O-methyltransferase inhibitors do not alter behaviour by elevating extracellular levels of free catecholamines levels but other explanations are needed.

Background c-Jun NH2-terminal kinases (JNKs) are strongly activated by a stressful cellular environment, such as chemotherapy and oxidative stress. Autophagy is a protein-degradation system in which double-membrane vacuoles called autophagosomes are formed. The autophagy-related gene Beclin 1 plays a key role in this process. We previously found that autophagy was induced by dihydroartemisinin (DHA) in pancreatic cancer cells. However, little is known about the complex relationship between ROS, JNK activation, autophagy induction, and Beclin 1 expression. Methods Cell viability and CCK-8 assays were carried out to determine the cell proliferation; small interfering RNAs (siRNAs) were used to knockdown c-Jun NH2-terminal kinases (JNK1/2) genes; western blot was performed to detect the protein expression of LC3, JNK, Beclin 1, caspase 3 and β-actin; production of intracellular ROS was analyzed using FACS flow cytometry; autophagy induction was confirmed by electron microscopy. Results In the present study, we explored the role of DHA and Beclin 1 expression in autophagy. DHA-treated cells showed autophagy characteristics, and DHA also activated the JNK pathway and up-regulated the expression of Beclin 1. Conversely, blocking JNK signaling inhibited Beclin 1 up-regulation. JNK activation was found to primarily depend on reactive oxygen species (ROS) resulting from the DHA treatment. Moreover, JNK pathway inhibition and Beclin 1 silencing prevented the induction of DHA-induced autophagy. Conclusions These results suggest that the induction of autophagy by DHA is required for JNK-mediated Beclin 1 expression.