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We sequenced the entire genome of isolates SMoV T1 and T2 of strawberry mottle virus (SMoV) from strawberry cultivar UC-5 grown in Tochigi, Japan. Homology and molecular phylogenetic analyses revealed that SMoV T1 and T2 are close homologs of previously reported Canadian and Czech strains. This is the first report describing the complete genome sequences of SMoV isolated in Japan and the construction of infectious SMoV clones. The stability of infectious clones in strawberry is discussed.

Strawberry vein banding virus (SVBV) and strawberry mild yellow edge virus (SMYEV) have caused disease in strawberries for many years in Japan, but little is known about their molecular properties. We thus determined the complete genomic sequence of an isolate of each virus from Tochigi Prefecture. A nucleotide sequence analysis showed that SVBV is closely related to Chinese isolates, whereas SMYEV is closely related to German and Argentine isolates. This is the first report describing the complete genome sequences of SVBV and SMYEV isolated in Japan. Moreover, we constructed an infectious SMYEV clone, which induced mild yellowing on inoculated strawberry plants.

Tomato infectious chlorosis virus (TICV) belongs to the genus Crinivirus in the family Closteroviridae. We used Agrobacterium co-infiltration leaf patch assays to investigate the suppression of RNA silencing by p27, which is encoded by TICV-RNA1 ORF2 in a genomic location similar to that of viral suppressor of RNA silencing (VSR) ORF in other criniviruses. Additionally, Nicotiana benthamiana was inoculated with a potato virus X vector (pGR106) carrying the p27 sequence (pGR106–p27) to examine whether p27 is a VSR. Plants infiltrated with pGR106–p27 exhibited more severe necrosis and mosaic symptoms. This is the first report describing a TICV VSR.

The complete nucleotide sequence of Tomato leaf curl New Delhi virus detected from melon in Indonesia-[Indonesia:Yogya:Melon:2015] (ToLCNDV-[ID:YG:Mel:15]) was determined. ToLCNDV-[ID:YG:Mel:15] has bipartite DNAs, and both DNAs showed highest identity to ToLCNDV-[Indonesia:Java:Cucumber:2008] (ToLCNDV-[ID:JV:Cuc:08]). However in a phylogenetic analysis based on AV1, ToLCNDV-[ID:YG:Mel:15] clustered with Squash leaf curl China virus (SLCCNV) isolates. Recombination analyses revealed that ToLCNDV-[ID:YG:Mel:15] is a recombinant between ToLCNDV and SLCCNV. Thus, the virus is a new recombinant of ToLCNDV. The infectivity of the recombinant ToLCNDV-[ID:YG:Mel:15] isolate was confirmed, and the inoculation induced mosaic on melon, but not on tomato, the primary natural host of ToLCNDV.

The soil-borne plasmodiophorid Polymyxa graminis is a vector for Barley yellow mosaic virus (BaYMV), which can severely damage barley plants. Although 22 disease resistance genes have been identified, only a few have been used for breeding virus-resistant cultivars. Recently, BaYMV strains capable of overcoming the effects of some of these genes have been detected. In this study, green fluorescent protein (GFP)-expressing BaYMV was constructed and used to examine viral dynamics in inoculated barley plants. Leaf inoculations resulted in higher infection rates than root or crown inoculations. Additionally, inoculations of some resistant cultivars produced infections that were similar to those observed in a field test. The results of this study indicate that the GFP-expressing virus is a useful tool for visualizing virus replication and dynamics, and for understanding resistance mechanisms.

Background: Patients with chronic obstructive pulmonary disease (COPD) are more susceptible to Aspergillus colonization or infection. Several studies have demonstrated that invasive pulmonary Aspergillosis (IPA) and Aspergillus hypersensitivity (AH) have a detrimental effect on COPD. However, it remains to be clarified whether Aspergillus colonization is associated with acute exacerbation of COPD (AECOPD). This study aimed to explore the impact of Aspergillus colonization in the lower respiratory tract on AECOPD. Method: Patients with Aspergillus colonization were identified from a retrospective cohort of hospitalized AECOPD from 2011 to 2016 in eight centers in Shanghai, China. The demographic information, conditions of the stable stage, clinical characteristics during hospitalization, and 1-year follow-up information after discharge were collected and compared to participants without fungi colonization. Result: Twenty-six hospitalized AECOPD patients with Aspergillus colonization and 72 controls were included in the final analysis after excluding patients with other fungi isolation and matching. The rates of recurrence of acute exacerbation within 90 days and 180 days after discharge in the patients with Aspergillus colonization were both significantly higher than that in the fungi negative patients (90 days: 19.2 vs. 4.2%, p = 0.029; 180 days: 23.1 vs. 4.2%, p = 0.010), and the all-cause mortality within 1 year was also higher (11.5 vs. 0.0%, p = 0.017). Multivariate logistic regression analysis showed that Aspergillus colonization was an independent risk factor for the recurrence of acute exacerbation within 90 days and 180 days (90 days: OR = 8.661, 95% CI: 1.496-50.159, p = 0.016; 180 days: OR =10.723, 95% CI: 1.936-59.394, p = 0.007). Conclusion: Aspergillus colonization may predict poor prognosis of AECOPD while leading to an increased risk of recurrent AECOPD in a short period.

H2O2 aerosol was used to oxidize dichlorvos vapor. 80.7% of dichlorvos, with the initial concentration of 90.9 mg/m3, was decontaminated when the H2O2 concentration kept in the range of 110~130 mg/m3 within 60 min. According to the oxidation products, analyzed by GC-MS, the decontamination reaction mechanism was discussed. Radical chain reaction is the main decontamination mechanism. H2O2 is dissociated to HOO-, which can induce the formation of ·OH. The double bond in dichlorvos molecule is attacked by ·OH to form molecule radical, which is further oxidized to 1,1-dichloro ethoxy dimethyl phosphate, 1,1,1-trichloro-2-hydroxyl-ethyl dimethyl phosphate, dimethyl phosphite, dimethyl phosphate, trimethyl phosphate, methyl phosphate, dichloro acetaldehyde, oxalic acid, CH2Cl2, CHCl3, parts of which are mineralized to phosphoric acid, CO2, H2O and chloridion.

To identify possible sites of viral attenuation, the complete nucleotide sequences of two isolates of Zucchini yellow mosaic virus (ZYMV) were determined; a severe isolate Z5-1 and an attenuated isolate from Z5-1 (designated ZYMV-2002). The viral genome of both isolates consisted of 9593 nucleotides in size and contained an open reading frame encoding a single polyprotein of 3080 amino acids. Comparison of the nucleotide sequences for Z5-1 and ZYMV-2002 revealed 14 nucleotide mutations, resulting in seven amino acid substitutions with four in the HC-Pro region, two in the CI region, and one in the NIb region. These results provide a genetic basis for future manipulation of the ZYMV reverse genetics system.

Dihydroartemisinin, a more water-soluble metabolite of artemisinin derivatives, is a safe and most effective antimalarial analog of artemisinin. In the present study, we investigated the antiangiogenic activity of dihydroartemisinin in vitro and in vivo, and investigated dihydroartemisinin-induced apoptosis in human umbilical vein endothelial cells (HUVEC). Dihydroartemisinin markedly reduced VEGF binding to its receptors on the surface of HUVEC. The expression levels of two major VEGF receptors, Flt-1 and KDR/flk-1, on HUVEC were lower following dihydroartemisinin treatment as shown by an immunocytochemical staining assay. The in vivo antiangiogenic activity was evaluated in the model of chicken chorioallantoic membrane (CAM) neovascularization. Dihydroartemisinin significantly inhibited CAM angiogenesis at low concentrations (5-30 nmol/100 microl per egg). We also investigated both qualitatively and quantitatively the induction of HUVEC apoptosis by dihydroartemisinin. A dose-related (5-80 microM) and time-dependent (6-36 h) increase in dihydroartemisinin-induced HUVEC apoptosis was observed by flow cytometry. Our results suggest that the antiangiogenic effect induced by dihydroartemisinin might occur by induction of cellular apoptosis and inhibition of expression of VEGF receptors. These findings and the known low toxicity of dihydroartemisinin indicate that it might be a promising candidate angiogenesis inhibitor.

A series of polypyridine Ruthenium(II) complexes Ru(II)(ttp)(L)X×PF 6 were synthesized and characterized, where L are 2,2′-bipyridine(bpy), 4,4′-dimethyl-2,2′-bipyridine(dmpy), 1,10-phenanthroline(phen) and X are Cl − , NCS − , CN − ; ttp=4′-p-tolyl-2,2′:6′,2′′-terpyridine. Their redox, spectroscopic and photophysical properties have been studied. All complexes have intense MLCT(d M →π L *) absorbance at approximately 500 nm and were found to be emitting with monoexponential decay of emission intensity at 298 K. The maximum wavelengths of absorbance and emission undergo a blue shift in the order of Cl − , NCS − , CN − . All complexes have an one-electron-oxidative process corresponding to Ru(III)/Ru(II) with a potential of about 0.8 V, and their oxidation potential increased also according to Cl − , NCS − , CN − . Two reversible or quasi-reversible one-electron-reductive processes based on ligands were also observed.

The photochemical behaviour of a squaraine dye ( 2 ( 4-ethylamino-2-hydroxyphenyl )-4 ( 4-e,hylaminopheny ) squaraine ( SQ ) ) in a TiO~ system was probed by UV-visible spectroscopy, fluorescence quenching measurements, single-photon counting and nanosecond fime-~ved laser flash photolysis. A strong interaction was observed between SQ and the TiO., colloidal particles, with an apparent associalion constam of 2.275 × IO s M - i. The fluorescence emission of the SQ dye was quenched by the TiOz colloid with an efficiency of 97%, r~ulting from electron transfer from the singlet excited state of the SQ dye to the conduction band of the semiconductor. The rate constant for the electron injection process was 1.97 × IO s s " t determined from the fluorescence lifetimes. Analysis of the transient absorption spectra confirmed the electron injection process from SQ to the TiO., p~u'ticles. © 1997 El~vier Science S.A.

Double-stranded RNA multimers of cucumber mosaic virus (CMV) satellite RNA were detected in CMV-infected plants. RT-PCR showed that plus-sense and minus-sense monomers and plus-sense multimers of satellite RNA were present. Multimeric minus-sense RNA was not present except in the form of multimeric dsRNA. Sequence analysis of 52 cloned junction regions in head-to-tail repeats of unit-length satellite RNA indicated that about 35% of the junction sequences were precise fusions of monomer units, 56% lacked sequence of the 5' component, and 10% lacked sequence of both 3' and 5' components. No junction contained additional nucleotides. Deletions at the junction regions may have accumulated during CMV multiplication in inoculated plants. These data suggest that replicase is not released from the template during synthesis of multimeric molecules of satellite RNA.

The transient phenomena in a laser flash photolysis study of two photosensitizing agents for photodynamic therapy, hypocrellin A(HA) and hypocrellin B(HB), are described in solvents of varying polarity at room temperature. The difference between the triplet- and ground-state absorption spectra of HA or HB shows peaks ca. 400, 530, 570 (sh) and 620 nm in Ar-saturated n-hexane solution. The absorption coefficients for the T–T absorption and triplet quantum yield of HA (or HB) have been determined. Electron transfer in the quenching of triplet HA or HB by ground-state molecules of the dye was investigated, and the bimolecular self-quenching rate constant for HA or HB in DMSO, which is 1.2×109 or 0.8×109 d mol-1 s-1 respectively, suggested a nearly diffusion-controlled quenching process of the triplet excited state. Meanwhile, the corresponding anion radicals HA-, HB-, which are produced by electron-transfer reactions from aromatic amines, have absorption maxima at 640 nm. Triplet-state lifetimes and electron-transfer kinetics are also reported.

Genotoxicity of aldicarb and methomyl was explored. The aldicarb and methomyl were diluted by the deionized water respectively, and then five concentrations of aldicarb were generated as 0.002, 0.02, 0.2, 2, 20 microg/L, methomyl as 0.02, 0.2, 2, 20, 200 microg/L. The micronucleus of carp erythrocyte was counted by micronucleus test. The mutation of bacteria was assessed by Ames test. The DNA damage of human lymphocytes was tested by comet assay. The genotoxicity of aldicarb and methomyl was estimated by the three toxicology tests mentioned above. The results showed that, in the micronucleus test, both any concentration of two pesticides were not able to induce higher frequency of micronucleus in carp erythrocyte (p0.05). Under condition of metabolic inactivation, although the number of colony with back mutation in any concentration of two pesticides did not exceed the double number of those with spontaneous mutation, the revertants of TA97 strains in the aldicarb 2-20 microg/L and the methomyl 20-200 microg/L were (129.17 +/- 17.00), (129.50 +/- 18.28), (109.83 +/- 10.80) and (114.17 +/- 9.37) entries/plate, respectively, they were significantly greater than those in spontaneous mutation (p0.05, p0.01). In the methomyl 200 microg/L group, the revertants of TA100 and TA102 strains were (147.83 +/- 23.29) and (275.83 +/- 20.63) entries/plate, respectively, they are significantly higher than that of the control group under condition of metabolic activation (p0.05). In comet assay, both the high concentration groups of aldicarb and methomyl resulted in different degrees of DNA damage of human peripheral blood lymphocytes. Compared with deionized water group, all of three indexes of comet assay in the aldicarb 20 microg/L groups and the methomyl 200 microg/L groups were significantly higher (p0.01). Despite that both aldicarb and methomyl did not results in damaging chromosome carp erythrocyte and producing apparent mutagenicity, the effect of mutagenicity and DNA damage in human lymphocytes were observed in high concentration groups of both aldicarb and methomyl. Water polluted by aldicarb and methomyl may have the potential adverse effects on the environment and human health.

To explore the role of bone marrow (BM) imprint in the diagnosis of hematological diseases.Between January 2002 and June 2008, a total of 3024 cases with BM smears, imprints and sections conducted simultaneously were recruited. There were 1667 males and 1357 females with a median age of 55 years old (range: 7 to 92). The cellularity on imprint and smear was evaluated with the standard cellularity on BM section. With the integrative diagnosis (including all examinations and clinical outcomes) as the standard, the diagnostic accuracy of hematological diseases were compared between BM imprint, smear and section groups. Another 79 cases of lymphoma and 114 cases of plasma cell myeloma (PCM) were selected for a correlation analysis of tumor cell infiltration patterns.BM imprint contained hematopoietic and non-hematopoietic regions and cells retained integrated structure. The cellularity evaluation by imprint was superior to smear overall. In BM imprint group, the diagnostic accuracy for hypersplenism (n = 130), metastatic carcinoma (n = 67), refractory anemia with excess blasts, myeloproliferative neoplasm (n = 174), and PCM (n = 94) were better than smear group (96.9% vs 80.7%, 91.0% vs 76.1%, 92.6% vs 81.5%, 92.5% vs 76.4%, and 97.8% vs 92.6% respectively, all P0.05); And the diagnostic accuracy for megaloblastic anemia (n = 69), acute myeloid leukemia (n = 104), refractory cytopenia with unilineage dysplasia (n = 15), refractory cytopenia with multilineage dysplasia (n = 22), and lymphoplasmacytic lymphoma (n = 12) were higher than biopsy section group (100% vs 84.0%, 91.3% vs 74.0%, 86.7% vs 60.0%, 90. 9% vs 72.7%, and 66.6% vs 50.0% respectively, all P0.05); And the diagnostic accuracy for myelodysplastic/myeloproliferative neoplasm (n = 26) was higher than smear group (76.3%, P0.05) and biopsy section group (78.2%, P0.05). Excellent correlations existed between BM imprint and section of the patients with lymphoma or with PCM (r = 0.90, r = 0.78, both P0.05).BM imprint contains the characteristics of both smear and section. BM imprint is superior to smear for an evaluation of cellularity. And it is also better than section for an analysis of cytological changes.

Cucumber mosaic virus satellite RNA (CARNA5) was used to investigate the usefulness of replicative form double-stranded RNAs as nonradioactive photobiotin-labelled probes in hybridization studies. In dot-blot hybridization using purified dsCARNA5 labelled with photobiotin as the probe, as little as 2pg of dsCARNA5 was detected. The sensitivity for detecting dsCARNA5 was higher on nylon membranes than on nitrocellulose filters, and dot-blot hybridization was more sensitive than gel electrophoresis with silver staining. CARNA5 in partially purified total nucleic acids extracted from CMV-infected tobacco leaves was readily detected by dot-blot hybridization. Preparations from healthy leaves gave no hybridization signal. In this study three types of CARNA5 were detected and separated by electrophoresis. The three types of CARNA5 hybridized with each other, however, the strongest hybridization signal was obtained with the homologous type of CA-RNA5.

Four satellite RNA isolates of cucumber mosaic virus (CMV), S19-, T43-, T62- and T73-sat-RNAs were obtained from CMV-infected plants collected in Tochigi Prefecture of Japan. Their physical and biological properties were investigated. T43- and T62-satRNAs induced necrosis on tomato like Y-satRNA, whereas S19- and T73-satRNAs did not. T43-satRNA was found to be identical to T62-satRNA by nucleotide sequence determination. The sequences were compared with each other and with other satellite RNAs, revealing high degree of nucleotide sequence homology between S19-satRNA and Y-satRNA (91%) and between T43-satRNA and OY2-satRNA (94%). Further analyses of the three sequences indicate that no common open reading frame is present and thus, they do not seem to encode any protein products essential for their biological activity. Competition of the satellite RNAs with Y-satRNA was examined by mixed inoculations under the condition using tobacco as a host and CMV-O as a helper virus. The relative replication ability of T43-satRNA is superior to that of Y-satRNA while T73-satRNA is apparently similar to or a little better than Y-satRNA.

Artesunate (ART), a semi-synthetic derivative of artemisinin extracted from the Chinese herb Artemisia annua, is a safe and effective antimalarial drug. In the present investigation, we analyzed the inhibitory effects of ART on angiogenesis and on VEGF production in chronic myeloid leukemia (CML) K562 cells in vitro and in vivo. In order to analyze the effect of ART on VEGF secretion in K562 cells, we examined the level of VEGF secreted in conditioned media (CM) by ELISA assay. The result showed that ART could decrease the VEGF level in CM of K562 cells, even at a lower concentration (2 micromol/l, P

To investigate the morphological changes of megakaryocytes with nuclear extrusion and nucleocytoplasmic separation, the morphological characteristics of megakaryocytes in peripheral blood films, bone marrow smears, and bone marrow biopsies from 4 newly diagnosed patients with primary myelofibrosis (PMF), myelodysplastic syndrome (MDS), myeloblastic leukemia with maturation (M(2)) and erythroleukemia (M(6)) were studied by using light microscope. The results showed that many kinds of dysmegakaryocytes were observed in bone marrow smears of 4 cases, while in case A (PMF) and case D (M(6)) micromegakaryocytes were ripped apart; in case B (MDS) and case C (M(2)) megakaryocytes were accompanied by nuclear extrusion or nucleocytoplasmic separation, and their bodies were large or giant, the part of nucleus separated from their body and little cytoplasm remained as micromegakaryocytes. The nucleocytoplasmic separation could be displayed by immunocytochemistry stain. It is concluded that the phenomenon of nuclear extrusion and nucleocytoplasmic separation in megakaryocytes suggested the process that dispersed multinuclear releasing towards surround or even totally left the cell body during the megakaryocyte maturation. It also showed that the micromegakaryocytes may be the result of nucleocytoplasmic separation or splittings from multi-separated nucleus.

Making use of the high expression efficiency of maltose-binding protein (MBP) in E. coli, the soluble and the inclusion body forms of MBP were purified to homogeneity by affinity chromatography and by denaturation, refolding and gel filtration respectively. After immunized Balb/C mice with two forms of MBP respectively, 5 clones for each antigen form were found to secrete monoclonal antibodies. The binding experiments showed that two antibodies obtained by the immunization with the refolded form of MBP had high affinity to the denatured MBP.