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Objective: To observe the effect of methylprednisolone (MP) plus adrenaline to treat patients injured by wasp stings., Methods: This study was carried out at The First College of Clinical Medical Science, China Three Gorges University, Yichang, Hubei Province, China, from June to November 2008. A total of 45 cases were divided into 2 groups. The MP was used alone in the control group, and MP plus adrenaline was used in the observation group. We compared the clinical effect between the 2 groups., Results: The alanine aminotransferase (ALT) (64.05 +/- 59.31 versus 124.14 +/- 80.01 U/L), and creatine kinase isoenzyme (CKMB) (90.32 +/- 85.19 versus 216.68 +/- 277.58 umol/L) levels of the observation group were significantly lower than those of the control group. The length of hospital stay of the observation group was significantly shorter than that of the control group (7.23 +/- 2.93 versus 11.23 +/- 8.02), while complications from the wasp sting were fewer than those of the control group. In addition, the level of peripheral blood leukocytes was positively correlated with the number of stings (r=0.733, p=0.001), levels of ALT and CKMB (r=0.627, p=0.001, and r=0.705, p=0.001), and length of hospital stay (r=0.667, p=0.001)., Conclusion: Allergic and inflammatory responses play an important role in addition to the direct effect of wasp venom on the human body. Compared with MP alone, early combination of adrenaline helps to further inhibit the diffusion of allergy and inflammatory cytokines, and therefore reduce the severity of injury.

Invadopodia are actin-based projections enriched with proteases, which invasive cancer cells use to degrade the extracellular matrix (ECM). The Phox homology (PX)-Src homology (SH)3 domain adaptor protein Tks5 (also known as SH3PXD2A) cooperates with Src tyrosine kinase to promote invadopodia formation but the underlying pathway is not clear. Here we show that Src phosphorylates Tks5 at Y557, inducing it to associate directly with the SH3-SH2 domain adaptor proteins Nck1 and Nck2 in invadopodia. Tks5 mutants unable to bind Nck show reduced matrix degradation-promoting activity and recruit actin to invadopodia inefficiently. Conversely, Src- and Tks5-driven matrix proteolysis and actin assembly in invadopodia are enhanced by Nck1 or Nck2 overexpression and inhibited by Nck1 depletion. We show that clustering at the plasma membrane of the Tks5 inter-SH3 region containing Y557 triggers phosphorylation at this site, facilitating Nck recruitment and F-actin assembly. These results identify a Src-Tks5-Nck pathway in ECM-degrading invadopodia that shows parallels with pathways linking several mammalian and pathogen-derived proteins to local actin regulation.

Pyramiding of genes that confer partial resistance is a method for developing wheat (Triticum aestivum L.) cultivars with durable resistance to leaf rust caused by Puccinia triticina. In this research, a doubled haploid population derived from the cross between the synthetic hexaploid wheat (SHW) (xAegilotriticum spp.) line TA4152-60 and the North Dakota breeding line ND495 was used for identifying genes conferring partial resistance to leaf rust in both the adult plant and seedling stages. Five QTLs located on chromosome arms 3AL, 3BL, 4DL, 5BL and 6BL were associated with adult plant resistance with the latter four representing novel leaf rust resistance QTLs. Resistance effects of the 4DL QTL were contributed by ND495 and the effects of the other QTLs were contributed by the SHW line. The QTL on chromosome arm 3AL had large effects and also conferred seedling resistance to leaf rust races MJBJ, TDBG and MFPS. The other major QTL, which was on chromosome arm 3BL, conferred seedling resistance to race MFPS and was involved in a significant interaction with a locus on chromosome arm 5DS. The QTLs and the associated molecular markers identified in this research can be used to develop wheat cultivars with potentially durable leaf rust resistance.

A total of 1,277 ESTs of Angiostrongylus cantonensis were downloaded from GenBank and analyzed with BlastX. SignalP V3.0 analysis was applied to predict potential putative antigen or allergen relative proteins with N-terminal secreted signal peptides or signal anchors. BlastX analysis showed that there were 614 ESTs scored more than 100, of which 14 were identical with A. cantonensis, 60 ESTs did not match any proteins in the databases. The identified 614 ESTs could be grouped into 10 categories, 80 ESTs expressed 22 antigen or allergen relative proteins, in which 12 had N-terminal secreted signal peptides and 3 had signal anchors.

Current durum wheat (Triticum turgidum L. subsp. durum (Desf.)) cultivars have little or no resistance to Fusarium head blight (FHB), a ravaging disease of cereal crops. A diploid wheatgrass, Lophopyrum elongatum (Host) A. Löve (2n = 2x = 14, EE genome), is an excellent source of FHB resistance. Through an extensive intergeneric hybridization using durum cultivar Langdon, we have developed a disomic alien addition line, named DGE-1 (2n = 28 + 2), with a wheatgrass chromosome pair. We used a unique method for isolating the addition line taking advantage of unreduced gametes functioning in Langdon x L. elongatum F1 hybrids in their first backcross to the Langdon parent, resulting in 35-chromosome plants from which we derived DGE-1. The addition line DGE-1 has a plant type similar to its Langdon parent, although it is shorter in height with narrower leaves and shorter spikes. It is meiotically and reproductively stable, generally forming 15 bivalents with two chiasmata each. The alien chromosome pair from the grass confers FHB resistance to the addition line, which has less than 21% infection on the visual scale, mean = 6.5%. Using various biochemical and molecular techniques (Giemsa C-banding, fluorescent genomic in situ hybridization (fl-GISH), chromosome-specific simple sequence repeat (SSR) markers, targeted region amplified polymorphism (TRAP) markers, and sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE)), we have shown that the extra chromosome involved is 1E of L. elongatum. This is the first time that FHB resistance has been discovered on chromosome 1E. We have established a chromosome-specific marker for 1E that may be used to screen fertile hybrid derivatives and durum addition lines for this chromosome that confers FHB resistance.

Stagonospora nodorum, causal agent of Stagonospora nodorum blotch (SNB), is a destructive pathogen of wheat worldwide. As is true for many necrotrophic host-pathogen systems, the wheat-S. nodorum system is complex and resistance to SNB is usually quantitatively inherited. We recently showed that S. nodorum produces at least four proteinaceous host-selective toxins that interact with dominant host sensitivity/susceptibility gene products to induce SNB in seedlings. Here, we evaluated a population of wheat recombinant inbred lines that segregates for Tsn1, Snn2, and Snn3, which confer sensitivity to the toxins SnToxA, SnTox2, and SnTox3, respectively, to determine if compatible host-toxin interactions are associated with adult plant susceptibility to SNB foliar disease under field conditions. Artificial inoculation of the population in 2 years and two locations with a fungal isolate known to produce SnToxA and SnTox2 indicated that compatible SnToxA-Tsn1 and SnTox2-Snn2 interactions accounted for as much as 18 and 15% of the variation in disease severity on the flag leaf, respectively. As previously reported for seedlings, the effects of these two interactions in conferring adult plant susceptibility were largely additive. Additional adult plant resistance QTLs were identified on chromosomes 1B, 4B, and 5A, of which, the 1B and 5A QTLs were previously reported to be associated with seedling resistance to SNB. Therefore, in this population, some of the same QTLs are responsible for seedling and adult plant resistance/susceptibility. This is the first report showing that host-selective toxins confer susceptibility of adult plants to SNB, further substantiating the importance of compatible toxin-host interactions in the wheat-S. nodorum pathosystem.

Resistance gene H26, derived from Aegilops tauschii Coss., is one of the most effective R genes against the Hessian fly [Mayetiola destructor (Say)], an important pest of wheat (Triticum aestivum L.). Using a limited number of PCR-based molecular markers a previous study mapped H26 to the wheat chromosomal deletion bin 3DL3-0.81-1.00. The objectives of this study were to saturate the chromosomal region harboring H26 with newly developed PCR-based markers and to investigate the collinearity of this wheat chromosomal region with rice (Oryza sativa L.) and Brachypodium distachyon genome. A population of 96 F(2) individuals segregating at the H26 gene locus was used for saturation mapping. All wheat ESTs assigned to the deletion bin 3DL3-0.81-1.00 were used to design STS (sequence tagged site) primers. The wheat ESTs mapped near H26 were further used to BLAST rice and B. distachyon genomic sequences for comparative mapping. To date, 26 newly developed STS markers have been mapped to the chromosomal region spanning the H26 locus. Two of them were mapped 1.0 cM away from the H26 locus. Comparative analysis identified genomic regions on rice chromosome 1 and Brachypodium Super contig 13 which are collinear with the genomic region spanning the H26 locus within the distal region of 3DL. The newly developed STS markers closely linked to H26 will be useful for mapped-based cloning of H26 and marker-assisted selection of this gene in wheat breeding. The results will also enhance understanding of this chromosomal region which contains several other Hessian fly resistance genes.

The Ogu cytoplasm for male sterility and its fertility restorer gene Rfo in canola (Brassica napus L.) were originally introgressed from radish (Raphanus sativus L.) and have been widely used for canola hybrid production and breeding. The objective of this study was to determine the physical location of the Rfo locus in the canola genome using fluorescence in situ hybridization and genetic mapping. For physical localization of the Rfo gene, two bacterial artificial chromosome (BAC) clones, G62 and B420, which were closely linked to the Rfo gene, were used as probes to hybridize with the somatic metaphase chromosomes of a canola hybrid variety, PHI-46 (46H02), containing the Rfo fragment. The results showed that both clones were physically located at the end of one large metacentric chromosome. By simultaneous use of two BAC clones and 45S rDNA repeated sequences as the probes, we demonstrated that the large metacentric chromosome probed with the two BAC clones did not carry 45S rDNA repeated sequences. The chromosome was 3.65 +/- 0.74 microm in average length (20 cells) and ranked second in size among the chromosomes without 45S rDNAs. The centromere index of the chromosome (20 cells) was calculated as 43.74 +/- 4.19. A comparison with previously reported putative karyotypes of B. napus (AACC) and its diploid ancestors Brassica rapa L. (AA) and Brassica oleracea L. (CC) suggests that the chromosome carrying the Rfo fragment might belong to one of three large metacentric chromosomes of the C genome. Genetic mapping has confirmed the localization of the Rfo fragment to the distal region of linkage group N19, which corresponds to the C genome in B. napus. This study has provided the evidence of the location of the Rfo gene on canola chromosomes and established a basic framework for further physical mapping and manipulation of the gene.

Tan spot, caused by Pyrenophora tritici-repentis, is a destructive foliar disease of wheat causing significant yield reduction in major wheat growing areas throughout the world. The objective of this study was to identify quantitative trait loci (QTL) conferring resistance to tan spot in the synthetic hexaploid wheat (SHW) line TA4152-60. A doubled haploid (DH) mapping population derived from TA4152-60 x ND495 was inoculated with conidia produced by isolates of each of four virulent races of P. tritici-repentis found in North America. QTL analysis revealed a total of five genomic regions significantly associated with tan spot resistance, all of which were contributed by the SHW line. Among them, two novel QTLs located on chromosome arms 2AS and 5BL conferred resistance to all isolates tested. Another novel QTL on chromosome arm 5AL conferred resistance to isolates of races 1, 2 and 5, and a QTL specific to a race 3 isolate was detected on chromosome arm 4AL. None of these QTLs corresponded to known host selective toxin (HST) insensitivity loci, but a second QTL on chromosome arm 5BL conferred resistance to the Ptr ToxA producing isolates of races 1 and 2 and corresponded to the Tsn1 (Ptr ToxA sensitivity) locus. This indicates that the wheat-P. tritici-repentis pathosystem is much more complex than previously thought and that selecting for toxin insensitivity alone will not necessarily lead to tan spot resistance. The markers associated with the QTLs identified in this work will be useful for deploying the SHW line as a tan spot resistance source in wheat breeding.

Objective: To observe effects of acupuncture at Neiguan (PC 6) on function of sinoatrial node, so as to provide experimental basis for clinical application of Neiguan (PC 6) to treatment of heart diseases., Methods: Fifty cases of heart diseases were randomly divided into 2 groups, a no-blocking group (n = 35) and a blocking group (n = 15). In the no-blocking group, sinoatrial node recovery time (SNRT), sinoatrial conduction time (SACT), sinoatrial node effective refractory period (SNERP) and heart rate (HR) were determined by using esophagus-left cardiac atrium regulating pulsation technique before and after acupuncture at Neiguan (PC 6); and in the blocking group, the vegetative nerve was blocked by intravenous injection of Propanolol and Atropine, and then SNRT, SACT, SNERP and intrinsic heart rate (IHR) were detected before and after acupuncture., Results: In the no-blocking group there were significant differences in SACT, SNERP and HR (all P < 0.05) and no significant difference in SNRT (P > 0.05) before and after treatment. In the blocking group, there were no significant differences in SNRT, SACT and SNERP and a significant difference in IHR before and after acupuncture (P < 0.05)., Conclusion: Acupuncture at Neiguan (PC 6) has a significant effect on function of sinoatrial node, and the mechanism is possibly related with the bidirectional regulative action of acupuncture at Neiguan (PC 6) on the autonomic nerve in the sinoatrial node.

Tan spot and Stagonospora nodorum blotch (SNB), caused by Pyrenophora tritici-repentis and Stagonospora nodorum, respectively, are two destructive foliar diseases of wheat, causing significant yield reduction worldwide. The objective of this study was to evaluate 172 accessions of wild emmer wheat (Triticum dicoccoides) for seedling resistance to tan spot and SNB. All accessions were inoculated with P. tritici-repentis race 1 and a mixture of three diverse isolates of S. nodorum, respectively. The accessions were also evaluated for sensitivity to host-selective toxins (HSTs), including ToxA produced by both S. nodorum and P. tritici-repentis and culture filtrate produced by S. nodorum. A total of 34 accessions were resistant to tan spot, and 136 accessions were resistant to SNB. Among these accessions, 31 were resistant to both diseases. Significant correlations between HST insensitivity and disease resistance were observed. Our results showed that T. dicoccoides is a good genetic source of resistance to tan spot and SNB in wheat.

One approach to understanding how orally administered drugs are absorbed and metabolized involves measuring compound concentrations in portal vein blood and in systemic circulation at various time points. In mice, blood samples are generally collected through terminal bleeding, a process that requires a large number of mice and is susceptible to variation between individuals. The authors developed a portal vein cannulation procedure for serial bleeding in the mouse, using a modified catheter containing a stainless steel stylet that is implanted directly in the portal vein. To demonstrate the technique, they orally administered two different compounds to mice and obtained blood samples from the tail vein and portal vein at different time points. They analyzed compound concentrations using liquid chromatography-tandem mass spectrometry. The technique refines existing methods for pharmacokinetic studies in the mouse and reduces the number of mice required.

Objective: To investigate the effects of 50 Hz magnetic fields (MF) on DNA double-strand breaks in human lens epithelial cells (hLECs)., Methods: The cultured human lens epithelial cells were exposed to 0.4 mT 50 Hz MF for 2 h, 6 h, 12 h, 24 h and 48 h. Cells exposed to 4-nitroquinoline-1-oxide, a DNA damage agent, at a final concentration of 0.1 micromol/L for 1 h were used as positive controls.After exposure, cells were fixed with 4 % paraformaldehyde and for H2AX (gamma H2AX) immunofluorescence measurement. gamma H2AX foci were detected at least 200 cells for each sample. Cells were classified as positive when more than three foci per cell were observed. Mean values of foci per cell and percentage of foci positive cells were adopted as indexes of DNA double-strand breaks., Result: The mean value of foci per cell and the percentage of gamma H2AX foci positive cells in 50 Hz MF exposure group for 24 h were (2.93 +/-0.43) and (27.88 +/-2.59)%, respectively, which were significantly higher than those of sham-exposure group [(1.77 +/-0.37) and (19.38+/-2.70)%, P <0.05], and the mean value of foci per cell and the percentage of gamma H2AX foci positive cells in 50 Hz MF exposure group for 48 h were (3.14 +/-0.35) and (31.00 +/-3.44)%, which were significantly higher than those of sham-exposure group (P <0.01). However there was no significant difference between 50 Hz MF exposure groups for 2 h, 6 h, 12 h and sham-exposure group for above two indexes (P >0.05)., Conclusion: 0.4 mT 50 Hz MF exposure for longer duration might induce DNA double-strand breaks in human lens epithelial cells in vitro.

Stem rust is a serious disease of wheat that has caused historical epidemics, but it has not been a threat in recent decades in North America owing to the eradication of the alternative host and deployment of resistant cultivars. However, the recent emergence of Ug99 (or race TTKS) poses a threat to global wheat production because most currently grown wheat varieties are susceptible. In this study, we evaluated a durum wheat-Aegilops speltoides chromosome translocation line (DAS15) for reaction to Ug99 and six other races of stem rust, and used molecular and cytogenetic tools to characterize the translocation. DAS15 was resistant to all seven races of stem rust. Two durum-Ae. speltoides translocated chromosomes were detected in DAS15. One translocation involved the short arm, centromere, and a major portion of the long arm of Ae. speltoides chromosome 2S and a small terminal segment from durum chromosome arm 2BL. Thus, this translocated chromosome is designated T2BL-2SL*2SS. Cytogenetic mapping assigned the resistance gene(s) in DAS15 to the Ae. speltoides segment in T2BL-2SL*2SS. The Ae. speltoides segment in the other translocated chromosome did not harbour stem rust resistance. A comparison of DAS15 and the wheat stocks carrying the Ae. speltoides-derived resistance genes Sr32 and Sr39 indicated that stem rust resistance gene present in DAS15 is likely novel and will be useful for developing germplasm with resistance to Ug99. Efforts to reduce Ae. speltoides chromatin in T2BL-2SL*2SS are currently in progress.

Objective: To observe the effects of different cell lysis buffers on protein quantification with Bradford method and bicinchoninic acid (BCA) method., Methods: Bradford method and BCA method were used to determine the concentration of bovine serum albumin (BSA) in different solutions (distilled water, cell lysis buffer used in two-dimensional differential in-gel electrophoresis and three kinds of cell lysis buffers used in conventional two dimensional gel electrophoresis), as well as the protein concentrations of cell lysates using these different lysis buffers. Bradford method was also applied to determine the protein concentrations of samples with repeated freeze thaw cycle, in different colorimetric cylinders, or using different standard curves from different periods., Result: The protein measurements increased for 1.2 to 2 fold when different cell lysis buffers were used in Bradford method, but the measurements increased with the increased concentration of BSA (r=0.989 approximately 0.996, P<0.05). For BCA, measurement reading increased about thousands times higher, even overflowed the limits of machine. Protein measurements didn't change significantly, only showed a declined trend after repeated freeze thaw cycle, while no significant changes were found using different colorimetric cylinders or standard curves from different periods., Conclusion: Bradford method may be the choice of the protein quantification in proteomics. However, optimization is required for specific experimental conditions.

Tan spot (caused by Pyrenophora tritici-repentis) and Stagonospora nodorum blotch (SNB) (caused by Stagonospora nodorum) are destructive fungal diseases of wheat (Triticum aestivum) throughout the world. Host plant resistance is thought to be an efficient and economical method of control. The objective of the present study was to identify novel sources of tan spot and SNB resistance in wheat genotypes derived from the crosses between wheat and alien species. Evaluations were conducted at the seedling stage in a growth chamber with 100% relative humidity. For each genotype, three replications were used for each disease. Among the 199 wheat-alien species derivatives evaluated, 65 exhibited resistance to tan spot and 30 showed resistance to SNB similar to BR34, a Brazilian wheat line used as the resistant control. Eleven derivatives were resistant to both diseases. Reactions of the derivatives and their respective wheat parents to tan spot and SNB suggest that resistance genes in the derivatives are derived from alien species. These derivatives can serve as desirable bridges for introgression of resistance genes from alien species to cultivated wheat, and could contribute novel and effective tan spot and SNB resistance to wheat breeding.

The cannabinoid-1 receptor (CB1R) has been implicated in the control of energy balance. To explore the pharmacological utility of CB1R inhibition for the treatment of obesity, we evaluated the efficacy of N-[(1S,2S)-3-(4-chlorophenyl)-2-(3-cyanophenyl)-1-methylpropyl]-2-methyl-2-[[5-(trifluoromethyl)pyridin-2-yl]oxy]propanamide (MK-0364) and determined the relationship between efficacy and brain CB1R occupancy in rodents. MK-0364 was shown to be a highly potent CB1R inverse agonist that inhibited the binding and functional activity of various agonists with a binding K(i) of 0.13 nM for the human CB1R in vitro. MK-0364 dose-dependently inhibited food intake and weight gain, with an acute minimum effective dose of 1 mg/kg in diet-induced obese (DIO) rats. CB1R mechanism-based effect was demonstrated for MK-0364 by its lack of efficacy in CB1R-deficient mice. Chronic treatment of DIO rats with MK-0364 dose-dependently led to significant weight loss with a minimum effective dose of 0.3 mg/kg (p.o.), or a plasma C(max) of 87 nM. Weight loss was accompanied by the loss of fat mass. Partial occupancy (30-40%) of brain CB1R by MK-0364 was sufficient to reduce body weight. The magnitude of weight loss was correlated with brain CB1R occupancy. The partial receptor occupancy requirement for efficacy was also consistent with the reduced food intake of the heterozygous mice carrying one disrupted allele of CB1R gene compared with the wild-type mice. These studies demonstrated that MK-0364 is a highly potent and selective CB1R inverse agonist and that it is orally active in rodent models of obesity.

Meiosis includes two successive divisions of the nucleus with one round of DNA replication and leads to the formation of gametes with half of the chromosomes of the mother cell during sexual reproduction. It provides a cytological basis for gametogenesis and nheritance in eukaryotes. Meiotic cell division is a complex and dynamic process that involves a number of molecular and cellular events, such as DNA and chromosome replication, chromosome pairing, synapsis and recombination, chromosome segregation, and cytokinesis. Meiosis maintains genome stability and integrity over sexual life cycles. On the other hand, meiosis generates genome variations in several ways. Variant meiotic recombination resulting from specific genome structures induces deletions, duplications, and other rearrangements within the genic and non-genic genomic regions and has been considered a major driving force for gene and genome evolution in nature. Meiotic abnormalities in chromosome segregation lead to chromosomally imbalanced gametes and aneuploidy. Meiotic restitution due to failure of the first or second meiotic division gives rise to unreduced gametes, which triggers polyploidization and genome expansion. This paper reviews research regarding meiosis-driven genome variation, including deletion and duplication of genomic regions, aneuploidy, and polyploidization, and discusses the effect of related meiotic events on genome variation and evolution in plants. Knowledge of various meiosis-driven genome variations provides insight into genome evolution and genetic variability in plants and facilitates plant genome research.

The discovery of novel acyclic amide cannabinoid-1 receptor inverse agonists is described. They are potent, selective, orally bioavailable, and active in rodent models of food intake and body weight reduction. A major focus of the optimization process was to increase in vivo efficacy and to reduce the potential for formation of reactive metabolites. These efforts led to the identification of compound 48 for development as a clinical candidate for the treatment of obesity.

The aneuploid stocks of durum wheat (Triticum turgidum L. subsp. durum (Desf.) Husnot) and common wheat (T. aestivum L.) have been developed mainly in 'Langdon' (LDN) and 'Chinese Spring' (CS) cultivars, respectively. The LDN-CS D-genome chromosome disomic substitution (LDN-DS) lines, where a pair of CS D-genome chromosomes substitute for a corresponding homoeologous A- or B-genome chromosome pair of LDN, have been widely used to determine the chromosomal locations of genes in tetraploid wheat. The LDN-DS lines were originally developed by crossing CS nulli-tetrasomics with LDN, followed by 6 backcrosses with LDN. They have subsequently been improved with 5 additional backcrosses with LDN. The objectives of this study were to characterize a set of the 14 most recent LDN-DS lines and to develop chromosome-specific markers, using the newly developed TRAP (target region amplification polymorphism)-marker technique. A total of 307 polymorphic DNA fragments were amplified from LDN and CS, and 302 of them were assigned to individual chromosomes. Most of the markers (95.5%) were present on a single chromosome as chromosome-specific markers, but 4.5% of the markers mapped to 2 or more chromosomes. The number of markers per chromosome varied, from a low of 10 (chromosomes 1A and 6D) to a high of 24 (chromosome 3A). There was an average of 16.6, 16.6, and 15.9 markers per chromosome assigned to the A-, B-, and D-genome chromosomes, respectively, suggesting that TRAP markers were detected at a nearly equal frequency on the 3 genomes. A comparison of the source of the expressed sequence tags (ESTs), used to derive the fixed primers, with the chromosomal location of markers revealed that 15.5% of the TRAP markers were located on the same chromosomes as the ESTs used to generate the fixed primers. A fixed primer designed from an EST mapped on a chromosome or a homoeologous group amplified at least 1 fragment specific to that chromosome or group, suggesting that the fixed primers might generate markers from target regions. TRAP-marker analysis verified the retention of at least 13 pairs of A- or B-genome chromosomes from LDN and 1 pair of D-genome chromosomes from CS in each of the LDN-DS lines. The chromosome-specific markers developed in this study provide an identity for each of the chromosomes, and they will facilitate molecular and genetic characterization of the individual chromosomes, including genetic mapping and gene identification.

Two synthetic hexaploid wheat lines (xAegilotriticum spp., 2n = 6x = 42, genomes AABBDD), SW8 and SW34, developed from the crosses of the durum wheat cultivar Langdon (Triticum turgidum L. var. durum, 2n = 4x = 28, genomes AABB) with two Aegilops tauschii Cosson accessions (2n = 2x = 14, genome DD), were determined to carry Hessian fly [Mayetiola destructor (Say)] resistance genes derived from the Ae. tauschii parents. SW8 was resistant to the Hessian fly biotype Great Plains (GP) and strain vH13 (virulent to H13). SW34 was resistant to biotype GP, but susceptible to strain vH13. Allelism tests indicated that resistance genes in SW8 and SW34 may be allelic to H26 and H13 or correspond to paralogs at both loci, respectively. H26 and H13 were localized to chromosome 4D and 6D, respectively, in previous studies. Molecular mapping in the present study, however, assigned the H26 locus to chromosome 3D rather than 4D. On the other hand, mapping of the resistance gene in SW34 verified the previous assignment of the H13 locus to chromosome 6D. Linkage analysis and physical mapping positioned the H26 locus to the chromosomal deletion bin 3DL3-0.81-1.00. A linkage map for each of these two resistance genes was constructed using simple sequence repeat (SSR) and target region amplification polymorphism (TRAP) markers.

Objective: To estimate the correlations between functional MRI (fMRI) parameters and the severity of chronic liver lesions of hepatitis B patients., Methods: 47 hepatitis B patients [6 with chronic hepatitis, 41 with cirrhosis (14 with Child-Pugh class A cirrhosis; 12 with class B cirrhosis; and 15 with class C cirrhosis)] and 10 normal volunteers, referred for measurements of apparent diffusion coefficient (ADC) values of the liver, perfusion imaging parameters, portal flow parameters and serum markers of hepatic fibrosis were included in the study. Diffusion-weighted imaging (DWI) with different b values and b value remainder was performed. Time to peak (TP), maximum slope of increase (MSI) and distribution volume (DV) were measured with dynamic contrast material-enhanced MR imaging. Portal velocity and portal flow with phase contrast (PC) were measured. The patients' serum hepatic fibrosis markers, including hyaluronic acid (HA), type-III-procollagen (PC III), laminin (LN) and type-IV-collagen (C IV), were measured and analyzed together with the fMRI results., Results: (1) The mean ADC3 in Child A, B, C cirrhosis patients was significantly lower than that in the controls (P < 0.05 in Child A, and P < 0.05 in Child B). (2) There was a significant increase of time to peak and a decrease of maximum slope of increase (P < 0.01) in the Child A, B, C patients than in the normal controls. (3) There was a significant decrease in portal velocity in cirrhotic patients as compared to that of the controls and chronic hepatitis patients (P < 0.01). (4) The mean HA in Child A, B, C cirrhosis patients was significantly higher than that in chronic hepatitis patients and in the controls (P < 0.01); The mean LN in Child A, B, C cirrhosis was also significantly higher than that in chronic hepatitis patients and in normal controls (P < 0.01); The mean PC III in Child A, B, C cirrhosis was significantly higher than that in the normal controls (P < 0.01)., Conclusion: fMRI parameters can reflect some changes of the livers, therefore fMRI parameters are of value in clinical diagnosis and treatment of chronic hepatitis B patients.

Four wheat (Triticum aestivum L.)-Thinopyrum ponticum derivatives SS5 (PI604926), SS156 (PI604947), SS363 (PI604970), and SS660 (PI604879), were identified as resistant to Fusarium head blight (FHB), a serious fungal disease of wheat worldwide. Seedling reactions to tan spot and Stagonospora nodorum blotch (SNB), two important foliar diseases of wheat, suggest that these four derivatives are resistant to tan spot and two of them (SS5 and SS156) are resistant to SNB. Fluorescent genomic in situ hybridization (FGISH) patterns of mitotic chromosomes indicate that these four derivatives are partial wheat-Th. ponticum amphiploids, each with a total of 56 chromosomes, though with different amounts of Th. ponticum chromatin. These four amphiploids were hybridized with each other to determine homology between the Th. ponticum genomes in each of the amphiploids. Analysis of chromosome pairing in the F1 hybrids using FGISH suggests that each amphiploid carries a similar set of Th. ponticum chromosomes. These wheat-Th. ponticum amphiploids represent a potential novel source of resistance to FHB, tan spot, and SNB for wheat breeding.

Hybrid necrosis is the gradual premature death of leaves or plants in certain F1 hybrids of wheat (Triticum aestivum L.), and it is caused by the interaction of two dominant complementary genes Ne1 and Ne2 located on chromosome arms 5BL and 2BS, respectively. To date, molecular markers linked to these genes have not been identified and linkage relationships of the two genes with other important genes in wheat have not been established. We observed that the F1 hybrids from the crosses between the bread wheat variety 'Alsen' and four synthetic hexaploid wheat (SHW) lines (TA4152-19, TA4152-37, TA4152-44, and TA4152-60) developed at the International Maize and Wheat Improvement Center (CIMMYT) exhibited hybrid necrosis. This study was conducted to determine the genotypes of TA4152-60 and Alsen at the Ne1 and Ne2 loci, and to map the genes using microsatellite markers in backcross populations. Genetic analysis indicated that Alsen has the genotype ne1ne1Ne2Ne2 whereas the SHW lines have Ne1Ne1ne2ne2. The microsatellite marker Xbarc74 was linked to Ne1 at a genetic distance of 2.0 cM on chromosome arm 5BL, and Xbarc55 was 3.2 cM from Ne2 on 2BS. Comparison of the genetic maps with the chromosome deletion-based physical maps indicated that Ne1 lies in the proximal half of 5BL, whereas Ne2 is in the distal half of 2BS. Genetic linkage analysis showed that Ne1 was about 35 cM proximal to Tsn1, a locus conferring sensitivity to the host selective toxin Ptr ToxA produced by the tan spot fungus. The closely linked microsatellite markers identified in this study can be used to genotype parental lines for Ne1 and Ne2 or to eliminate the two hybrid necrosis genes using marker-assisted selection.

Luminescent 20-nm silicon dioxide nanoparticles containing dibromofluorescein (D; particles denoted D-SiO(2)) were synthesized by the sol-gel method. In the presence of Pb(Ac)(2) as a heavy atom perturber, the particle can emit the intense and stable room temperature phosphorescence(RTP) signal on a polyamide membrane at the wavelength of lambdaex(max)/lambdaem(max) = 460/623 nm for D. Our research indicates that the specific immune reaction between goat-anti-human IgG antibody labeled with D-SiO(2) and human IgG can be carried out quantitatively on a polyamide membrane, and the phosphorescence intensity was evidently enhanced after the immunoreaction. Thus a new solid substrate-room temperature phosphorescence immunoassay (SS-RTP-IA) for determination of human IgG was established. The linear range of this method is 0.0624-20.0 pg spot(-1) of human IgG (corresponding to a concentration range of 0.156-50.0 ng ml(-1), sample volume: 0.40 microl spot(-1)). The regression equation of the working curve is DeltaIp = 94.39 + 17.00 m IgG (pg spot(-1)) (460/623 nm, r = 0.9998). Detection limit calculated as 3 Sb/k is 0.015 pg spot(-1). After elevenfold replicate measurement, RSD are 3.2% and 2.4% for samples containing 0.156 and 50.0 ng l(-1) IgG, respectively. This method is sensitive, accurate, and of high precision. And it has been applied to the determination of IgG in human serum with satisfactory results. Meanwhile, the mechanism of SS-RTP-IA based on an antibody labeled with nanoparticles containing dibromofluorescein luminescent molecules was discussed.

Two durum wheat [Triticum turgidum L. ssp. durum (Desf.) Husn.] lines carrying the high-molecular-weight (HMW) glutenin subunits (GS) 1 D x 5 + 1Dy10 encoded by Glu-D1d, L252 and S99B34, were characterized using fluorescent genomic in-situ hybridization (FGISH) and microsatellite markers. These two durum lines were derived from the crosses in which the common wheat (T. aestivum L.) 'Len' and durum wheat 'Langdon' (LDN) and 'Renville' were involved. FGISH patterns of the mitotic chromosomes indicated that these two durum lines have one pair of 1AS.1AL-1DL translocated chromosomes in which the terminal region of 1AL was replaced by a homoeologous segment of 1DL. The 1DL segment spans approximately 31% of the long arm of the translocated chromosome. Microsatellite marker analysis confirmed the 1AS.1AL-1DL translocation and determined the translocation breakpoint to be distal to Xgwm357 on 1AL. Seed storage proteins (GS and gliadins) were analysed in these two 1AS.1AL-1DL translocation lines and three sib lines (L092, S99B19 and S99B33) using SDS-PAGE and A-PAGE. The SDS-PAGE and A-PAGE profiles demonstrated that the two low yielding lines (L252 and S99B19) had the low-molecular-weight (LMW) -1 GS encoded by Glu-A3k and Glu-B3s and 1B-encoded gliadins from LDN, and the other three lines (L092, S99B33 and S99B34) with higher yield had LMW-2 GS and 1B-encoded gliadins from Renville, suggesting that undesirable genetic components from LDN might limit substantial improvement of yield. Thus, the translocation lines with 1 D x 5 + 1Dy10 and LMW-2, which are associated with good bread-making and pasta qualities, respectively, in a good genetic background will be useful for developing durum cultivars with dual-purpose end-use. Results from this study demonstrate that the D-genome could play an important role in the genetic improvement of durum wheat and evolution of the A- and B-genomes in tetraploid wheat.

Aim: To study the inhibitory effect of transfected PTEN on LoVo cells., Methods: Human PTEN cDNA was transferred into LoVo cells via lipofectin and PTEN mRNA levels and its expression were analyzed by Western blot and flow cytometry. Before or after transfection, the effects of 5-Fu on inhibiting cell proliferation and inducing apoptosis were measured by flow cytometry, DNA bands and MTT., Results: PTEN transfection significantly up-regulated PTEN expression in LoVo cells. 5-Fu inhibited cell proliferation and induced apoptosis in transfected LoVo cells., Conclusion: Transfected PTEN can remarkably up-regulate PTEN expression in LoVo cells and promote the apoptosis. PTEN transfection is associated with 5-Fu treatment effect and has a cooperatively cytotoxic effect.

The glutenin and gliadin proteins of wild emmer wheat, Triticum turgidum L. var. dicoccoides, have potential for improvement of durum wheat ( T. turgidum L. var. durum) quality. The objective of this study was to determine the chromosomes controlling the high molecular weight (HMW) glutenin subunits and gliadin proteins present in three T. turgidum var. dicoccoides accessions (Israel-A, PI-481521, and PI-478742), which were used as chromosome donors in Langdon durum- T. turgidum var. dicoccoides (LDN-DIC) chromosome substitution lines. The three T. turgidum var. dicoccoides accessions, their respective LDN-DIC substitution lines, and a number of controls with known HMW glutenin subunits were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), urea/SDS-PAGE, and acid polyacrylamide gel electrophoresis (A-PAGE). The results revealed that all three T. turgidum var. dicoccoides accessions possess Glu-A1 alleles that are the same as or similar to those reported previously. However, each T. turgidum var. dicoccoides accession had a unique Glu-B1 allele. PI-478742 had an unusual 1Bx subunit, which had mobility slightly slower than the 1Ax subunit in 12% SDS-PAGE gels. The subunits controlled by chromosome 1B of PI-481521 were slightly faster in mobility than the subunits of the Glu-B1n allele, and the 1By subunit was identified as band 8. The 1B subunits of Israel-A had similar mobility to subunits 14 and 16. The new Glu-B1 alleles were designated as Glu-B1be in Israel-A, Glu-B1bf in PI-481521, and Glu-B1bg in PI-478742. Results from A-PAGE revealed that PI-481521, PI-478742, and Israel-A had eight, 12, and nine unique gliadin bands, respectively, that were assigned to specific chromosomes. The identified glutenin subunits and gliadin proteins in the LDN-DIC substitution lines provide the basis for evaluating their effects on end-use quality, and they are also useful biochemical markers for identifying specific chromosomes or chromosome segments of T. turgidum var. dicoccoides.

Background: To clarify the features of gene variation among epidemic strains of human rotavirus NSP4 in China., Methods: SP4 genes from 27 epidemic strains of human rotavirus isolated in different area of China in recent years were amplified with RT-PCR, the resulted cDNAs were cloned and sequenced. The sequences of full length cDNAs were compared with 10 rotavirus NSP4 sequences available in the GenBank using the Clustal x 1.8 TreeView32 and DNA Star softwares. The G serotype of VP7 was analysed by PCR., Results: The homology of the amino acid among the 27 rotavirus strains isolated in China was 81.7%-99.4%. Based on the variation of amino acid sequence, the virus strains can be divided into two groups, represented by Wa and KUN with the homology of 92.0%-99.4% and 92.0%-98.9% within each group, respectively. The diversity between the two groups were 16.6%-21.0%. The Wa group could further be separated into three subgroups, according to the diversity between those strains and the characterization in the highly variable domain. The association between VP7 serotype and NSP4 genotype was not strong., Conclusions: The NSP4 gene of human rotavirus epidemic strains in China can be divided into Wa and KUN two groups, Wa group is the main group and contains three subgroups possessing characteristic amino acid sites. Samples isolated in the same year but not in the same area shared higher homology.

In recent decades, protected-area management in many developing countries has been molded to win the support and participation of local people. Increasingly, research initiatives are undertaken to enhance understanding of the perceptions and attitudes of rural stakeholders. The effectiveness of the expanding protected-area system in China is critically constrained by similar considerations of community mentality. This paper provides an empirical assessment of local responses towards conservation efforts based on a case study of the recently established Shimentai Nature Reserve (SNR) situated in Yingde, Guangdong Province, China. Questionnaire surveys, face-to-face interviews, and group discussions were employed to gauge local residents' knowledge, perceptions, and expectations towards the SNR. The study covered seven villages situated in and around the reserve, grouped into far-zone (floodplain) and near-zone (upland) categories, accommodating Hakka (Han) and minority Yao peoples. Many respondents had inadequate knowledge, understanding, and perceptions of the reserve. Although most respondents welcomed the expected park-related dividends, the most affected near-zone villagers anticipated losses due to restriction on traditional resource-extraction activities in the forest. The local expectations were influenced by place of residence, emigration of rural young, and household affluence. The lack of local participation in management and inadequate dissemination of information posed obstacles to effective conservation. The high expectation of accruing benefits from development projects, including tourism, might be misplaced and might not be realized. The present honeymoon period of positive attitudes could end soon if the hopes remain unfulfilled for too long. The need to win sustained local support and provide alternative means of livelihood is emphasized.

To further reveal the risks of heroin abuse to human body, and to determine the injuries of oxidation, peroxidation and lipoperoxidation induced by nitric oxide and other free radicals to heroin abusers, we determined and compared plasma values of lipoperoxides (LPO), nitric oxide (NO), vitamin C (VC), vitamin E (VE), beta-carotene (beta-CAR) and erythrocyte values of LPO, superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) in 114 heroin abusers and 100 healthy volunteers. Using linear regression and correlation as well as stepwise regression and correlation, we also analyzed the effect of the abusing duration, and daily abusing quantity on the above-mentioned biochemical parameters in the heroin abusers. The results showed that, compared with the healthy volunteer groups, the average plasma values of LPO, and NO, and the average erythrocyte value of LPO in the heroin abuser group were significantly increased (P < 0.0001), and the average plasma values of VC, VE, and beta-CAR and the average erythrocyte values of SOD, CAT, and GSH-Px were significantly decreased (P < 0.0001). Analysis of linear regression and correlation showed that with prolonged heroin abusing and with increased daily quantity in the heroin abusers, the plasma values of LPO, and NO, and the erythrocyte value of LPO were gradually increased (P < 0.001), whereas the plasma values of VC, VE, and beta-CAR and the erythrocyte values of SOD, CAT, and GSH-Px were gradually decreased (P < 0.001). Analysis of stepwise regression and correlation indicated that the plasma values of NO, VC and VE were closely correlated with the abusing duration and daily abusing quantity. These results indicate that the balance between oxidation and antioxidation in the heroin abusers was seriously disturbed, and the injuries induced by nitric oxide and other free radicals, through oxidation, peroxidation and lipoperoxidation to the bodies of heroin abusers exacerbated. It is therefore necessary that in abstaining from heroin dependence, the heroin abusers should acquire sufficient quantities of antioxidants such as VC, VE and beta-CAR.

Aim: To study the efficacy of huperzine-A capsules (Hup) on memory and learning performance of adolescent students., Methods: Using double-blind and matched pair method, 34 pairs of junior middle school students complaining of memory inadequacy were divided into two groups by normal psychological health inventory (PHI), similar memory quotient (MQ), same sex and class. The Hup group was administrated orally 2 capsules of Hup (each contains Hup 50 micrograms) b.i.d., and the placebo group was given 2 capsules of placebo (starch and lactose inside) b.i.d. for 4 wk., Results: At the end of trial, the Hup group's MQ (115 +/- 6) was more than that of the placebo group (104 +/- 9, P < 0.01), and the scores of Chinese language lesson in the Hup group were elevated markedly too., Conclusion: The Hup capsules enhance the memory and learning performance of adolescent students.

Aim: To compare the efficacy and safety between huperzine-A (Hup) in capsules and tablets for treating patients with Alzheimer disease (AD)., Methods: Using multicenter, prospective, double-blind, double-mimic, parallel, positive controlled and randomized methods, 60 patients meeting with the NINCDS-ARDRA criteria of AD were divided into 2 equal groups. Patients in the capsule group received 4 capsules of Hup (each contains 50 micrograms) and 4 tablets of placebo (lactose and starch inside); while the tablet group received 4 tablets of Hup (each contains 50 micrograms) and 4 capsules of placebo, p.o., twice a day for 60 d. All the patients were evaluated with a lot of related ranting scales, and physiological and laboratory examination., Results: There were significant differences (P < 0.01) on all the psychological evaluations between 'before' and 'after' the 60-d trial of 2 groups, but there was no significant difference between 2 groups by group t test (P > 0.05). The changes of oxygen free radicals in 2 groups showed marked improvement. No severe side effect besides moderate to mild nausea was found in both groups., Conclusion: There is equal efficacy and safety between Hup in capsule and tablet for treating patients with AD, and Hup can reduce the pathological changes of the oxygen free radicals in the plasma and erythrocytes of patients with AD.

Aim: To evaluate the efficacy and safety of tablet huperzine-A (Hup) in patients with Alzheimer's disease., Methods: Using multicenter, prospective, double-blind, parallel, placebo controlled and randomized method, 50 patients were administered orally 0.2 mg (4 tablets) Hup and 53 patients were given po 4 tablets of placebo bid for 8 wk. All patients were evaluated with Wechsler memory scale, Hasegawa dementia scale, mini-mental state examination scale, activity of daily living scale, treatment emergency symptom scale, and measured with BP, HR, ECG, EEG, ALT, AKP, BUN, Cr, Hb, WBC, and urine routine., Results: About 58% (29/50) of patients treated with Hup showed improvements in their memory (P < 0.01), cognitive (P < 0.01), and behavioral (P < 0.01 functions. The efficacy of Hup was better than placebo (36%, 19/53) (P < 0.05). No severe side effect was found., Conclusion: Hup is a promising drug for symptomatic treatment of Alzheimer's disease.