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Pyrimethanil myristic salt was synthesized and its heat capacities were measured with an automated adiabatic calorimeter over the temperature range from T=(79 to 360) K.The melting point, molar enthalpy, ΔfusHm and entropy, ΔfusSm, of fusion of this compound were determined to be (321.84 ± 0.05) K, (56.53 ± 0.03) kJ · mol−1 and (175.64 ± 0.05) J · mol−1 · K−1, respectively. The purity of the compound was calculated to be 98.99 mol% by using the fractional melting technique. The thermodynamic functions relative to the reference temperature, T=298.15 K, were calculated based on the heat capacity measurements in the temperature ranges from T=(80 to 360) K. The TG-DTG results demonstrate that the mass loss of the sample takes place in one step with the maximum rate at T=500 K, which was caused by evaporation of the sample.

The molar heat capacities of 1-(2-hydroxy-3-chloropropyl)-2-methyl-5-nitroimidazole (Ornidazole) (C7H10CIN3O3) with purity of 99.72mol% were measured with an adiabatic calorimeter in the temperature range between 79 and 380K. The melting-point temperature, molar enthalpy Delta(fus)H(m), and entropy, Delta(fus)S(m), of fusion of this compound were determined to be 358.59 +/- 0.04K, 21.38 +/- 0.02 kJ mol(-1) and 59.61 +/- 0.05 J K-1 mol(-1), respectively, from fractional melting experiments. The thermodynamic function data relative to the reference temperature (298.15 K) were calculated based on the heat capacities measurements in the temperature range from 80 to 380 K. The thermal stability of the compound was further investigated by DSC and TG. From the DSC curve an intensive exothermic peak assigned to the thermal decomposition of the compound was observed in the range of 445-590 K with the peak temperature of 505 K. Subsequently, a slow exothermic effect appears when the temperature is higher than 590 K, which is probably due to the further decomposition of the compound. The TG curve indicates the mass loss of the sample starts at about 440K, which corresponds to the decomposition of the sample. (C) 2003 Elsevier B.V. All rights reserved.

Low-temperature heat capacities of penconazole (C 13 H 15 Cl 2 N 3 ) were precisely measured with an automated adiabatic calorimeter over the temperature rang from 78 to 364 K. The sample was observed to melt at 332.38±0.06 K. The molar enthalpy and entropy of fusion of the compound were determined to be 33580±11 J mol −1 , 101.03±0.02 J mol −1 K −1 , respectively. Further research of the melting process for this compound was carried out by means of differential scanning calorimetry (DSC) technique. The result was in agreement with that obtained from the measurements of heat capacities.

Carboxin was synthesized and its heat capacities were measured with an automated adiabatic calorimeter over the temperature range from 79 to 380K. The melting point, molar enthalpy (Delta(fus)H(m)) and entropy (Delta(fus)S(m)) of fusion of this compound were determined to be 365.29 +/- 0.06K, 28.193 +/- 0.09 kJ mol(-1) and 77.180 +/- 0.02 J mol(-1) K-1, respectively. The purity of the compound was determined to be 99.55 mol% by using the fractional melting technique. The thermodynamic functions relative to the reference temperature (298.15 K) were calculated based on the heat capacity measurements in the temperature range between 80 and 360 K. The thermal stability of the compound was further investigated by differential scanning calorimetry (DSC) and thermogravimetric (TG) analysis. The DSC curve indicates that the sample starts to decompose at ca. 290degreesC with the peak temperature at 292.7degreesC. The TG-DTG results demonstrate the maximum mass loss rate occurs at 293degreesC corresponding to the maximum decomposition rate. (C) 2003 Elsevier B.V All rights reserved.

Low-temperature heat capacities of the 9-fluorenemethanol (c14h12o) have been precisely measured with a small sample automatic adiabatic calorimeter over the temperature range between t = 78 k and t = 390 k. the solid-liquid phase transition of the compound has been observed to be t-fus = (376.567 +/- 0.012) k from the heat-capacity measurements. the molar enthalpy and entropy of the melting of the substance were determined to be delta(fus)h(m) = (26.273 +/- 0.013) kj (.) mol(-1) and delta(fus)s(m) = (69.770 +/- 0.035) j (.) k-1 (.) mol(-1). the experimental values of molar heat capacities in solid and liquid regions have been fitted to two polynomial equations by the least squares method. the constant-volume energy and standard molar enthalpy of combustion of the compound have been determined, delta(c)u(c14h12o, s) = -(7125.56 +/- 4.62) kj (.) mol(-1) and delta(c)h(m)degrees(c14h12o, s) = -(7131.76 +/- 4.62) kj (.) mol(-1), by means of a homemade precision oxygen-bomb combustion calorimeter at t = (298.15 +/- 0.001) k. the standard molar enthalpy of formation of the compound has been derived, delta(f)h(m)degrees (c14h12o, s) = -(92.36 +/- 0.97) kj (.) mol(-1), from the standard molar enthalpy of combustion of the compound in combination with other auxiliary thermodynamic quantities through a hess thermochemical cycle. (c) 2004 elsevier ltd. all rights reserved.

Transcriptional activation is associated commonly with recruitment of histone acetylases, while repression involves histone deacetylases (HDACs). Here, we provide evidence to suggest that STAT5 activates gene expression by recruiting HDAC. The interleukin-3 (IL-3)-dependent expression of the Id-1 gene, encoding a helix-loop-helix (HLH) transcriptional inhibitor, is activated by both C/EBPbeta and STAT5 transcription factors bound to its pro-B-cell enhancer (PBE), but is inhibited by HDAC inhibitors in Ba/F3 cells. STAT5 interacts with HDAC1 in the PBE region, resulting in deacetylation of histones, as well as C/EBPbeta, whose acetylation diminishes its DNA-binding activity. Consistently, expression of an acetylation-resistant mutant of C/EBPbeta results in IL-3-independent expression of the Id-1 gene. Thus, we propose a novel mechanism by which STAT5 mediates the deacetylation of C/EBPbeta, allowing transcriptional activation.

E2A and HEB are basic helix-loop-helix transcription factors essential for T cell development. Complete inhibition of their activities through transgenic overexpression of their inhibitors Id1 and Tal1 leads to a dramatic loss of thymocytes. Here, we suggest that bHLH proteins play important roles in establishing thresholds for pre-TCR and TCR signaling. Inhibition of their function allows double-negative cells to differentiate without a functional pre-TCR, while anti-CD3 stimulation downregulates bHLH activities. We also find that the transcription factor NF-kappaB becomes activated in transgenic thymocytes. Further activation of NF-kappaB exacerbates the loss of thymocytes, whereas inhibition of NF-kappaB leads to the rescue of double-positive thymocytes. Therefore, we propose that E2A and HEB negatively regulate pre-TCR and TCR signaling and their removal causes hyperactivation and apoptosis of thymocytes.

Dendritic cells (DC) comprise plasmacytoid DC (pDC) and conventional DC (cDC) subsets; the latter is further separated into type 1 (cDC1) and type 2 (cDC2). These cells play a role in initiating and modulating immune responses. Their development is controlled by a network of transcription factors including E proteins consisting of E12, E47, HEB and E2-2, which share a basic helix-loop-helix domain mediating dimerization and DNA binding. Their activity is regulated by their antagonists, Id1-4. The balance between E and Id proteins regulates the transcription of E protein target genes. While E2-2 is essential for pDC differentiation and Id2 is indispensable for cDC1 formation, little is known about how the balance between E and Id proteins controls the early steps of the commitment of common DC precursors (CDP) to the diverse DC subsets. To address this, we used a mouse model that inducibly expresses a chimeric protein, ET2, which contains the DNA binding domain of Tal1 and the transactivation domains of E47. It can compete with Id proteins to bind endogenous E proteins and restore their DNA binding activity, thus increasing net E protein activity. Our data indicated that ET2 expression inhibited the differentiation of pDC and cDC2 subsets in vivo and in vitro. In contrast, cDC1 production was slightly elevated in steady state but dramatically increased in allergic reactions. Furthermore, we detected an augmented proportion of the newly identified cDC1 precursors and decreased representation of cDC2 precursors, suggesting E proteins skew CDPs toward the cDC1 lineage. These results are consistent with reports that E proteins activate IRF8 gene transcription. Further investigation is in progress to elucidate the molecular mechanisms.

Innate lymphoid cells (ILCs) have gained recognition as important regulators in various immune responses. Current paradigm states that all ILCs originate from common lymphoid progenitors (CLP) in the bone marrow through an array of downstream ILC progenitors. Id2, a member of the Id family that inhibits E protein transcription factors, is expressed in these ILC progenitors and is indispensable for ILC differentiation. Interestingly, we found that down-regulating E protein function in the thymus by Id1 overexpression or deletion of two E protein genes resulted in massive expansion of ILC2s in the thymus as well as other organs where ILC2 normally reside. The phenotypic thymic ILC2s expressed ILC2 signature genes and down-regulated T lineage-specific genes. Moreover, Id1 transgenic mice exhibited spontaneous infiltration of eosinophils and heightened type 2 immune responses to the allergen, papain, in the lung. Furthermore, these mice had increased ability in the expulsion of helminth parasites, N. Brasiliensis. These results raise an interesting question as to whether the thymus is a natural site for ILC2 production and the phenotypes of the mutant mice simply highlight such capacity. Indeed, we found that wild type DN1 and DN3 thymocytes can differentiate into IL-5 and IL-13-producing ILC2 cells in vitro. Ablation of E proteins in DN3 cells accelerated the differentiation while gain of E protein function inhibited the process. Together, our data reveal a new ILC2 differentiation program occurring in the thymus and a critical role of E proteins in restraining such a differentiation potential while ensuring proper T cell production.

The family of inhibitor of differentiation (Id) proteins is a group of evolutionarily conserved molecules, which play important regulatory roles in organisms ranging from Drosophila to humans. Id proteins are small polypeptides harboring a helix-loop-helix (HLH) motif, which are best known to mediate dimerization with other basic HLH proteins, primarily E proteins. Because Id proteins do not possess the basic amino acids adjacent to the HLH motif necessary for DNA binding, Id proteins inhibit the function of E protein homodimers, as well as heterodimers between E proteins and tissue-specific bHLH proteins. However, Id proteins have also been shown to have E protein-independent functions. The Id genes are broadly but differentially expressed in a variety of cell types. Transcription of the Id genes is controlled by transcription factors such as C/EBPβ and Egr as well as by signaling pathways triggered by different stimuli, which include bone morphogenic proteins, cytokines, and ligands of T cell receptors. In general, Id proteins are capable of inhibiting the differentiation of progenitors of different cell types, promoting cell-cycle progression, delaying cellular senescence, and facilitating cell migration. These properties of Id proteins enable them to play significant roles in stem cell maintenance, vasculogenesis, tumorigenesis and metastasis, the development of the immune system, and energy metabolism. In this review, we intend to highlight the current understanding of the function of Id proteins and discuss gaps in our knowledge about the mechanisms whereby Id proteins exert their diverse effects in multiple cellular processes.

Series of isoxazole derivatives were synthesized by substituted chalcones and 2-chloro-6-fluorobenzene formaldehyde oxime with 1,3-dipolar cycloaddition. The target compounds were determined by melting point, IR, 1H NMR, elemental analyses and HRMS. The crystal structure of compound 3a was detected by X-ray diffraction and it crystallizes in the triclinic space group p2(1)/c with z = 4. The molecular geometry of compound 3a was optimized using density functional theory (DFT/B3LYP) method with the 6-31G+(d,p) basis set in the ground state. From the optimized geometry of the molecule, FT-IR, FT-Raman, HOMO–LUMO and natural bond orbital (NBO) were calculated at B3LYP/6-31G+(d,p) level. Finally, the antifungal activity of the synthetic compounds were evaluated against Pythium solani, Gibberella nicotiancola, Fusarium oxysporium f.sp. niveum and Gibberella saubinetii.

This study aims to analyse the value of CODEHOP RT-PCR in the detection of Flavivirus. According to the amino acid sequences of polyproteins of different flaviviruses published in GenBank, a pair of primers was designed using the CODEHOP method. One-step RT-PCR was used to detect Japanese encephalitis virus strain JEV1201, Dengue virus strain JKD001, and yellow fever virus vaccine YV6161. BLAST analysis and phylogenetic analysis were performed after the RT-PCR products of nucleocapsid genes were sequenced. The results showed that this method could amplify Flavivirus specifically, and the size and sequence of the target fragment accorded with the anticipated result. JEV1201 had the highest homology to Japanese encephalitis virus strain YL2009-4/YC2009-3, belonging to the branch of the phylogenetic tree of Japanese encephalitis virus strains. JKD001 had the highest homology to Dengue virus strain DENV-2/ID/1022DN/1975, belonging to the branch of the phylogenetic tree of Dengue virus strains. YV6161 had the highest homology to Yellow fever virus strain 17D, belonging to the branch of the phylogenetic tree of Yellow fever virus strains. In conclusion, the method of CODEHOP RT-PCR can be effectively used to detect, identify, and phylogenetically analyse Flavivirus.

T regulatory (Treg) cells play crucial roles in the regulation of cellular immunity. The development of Treg cells depends on signals from TCRs and IL-2Rs and is influenced by a variety of transcription factors. The basic helix-loop-helix proteins are known to influence TCR signaling thresholds. Whether this property impacts Treg differentiation is not understood. In this study, we interrogated the role of basic helix-loop-helix proteins in the production of Treg cells using the CD4 promoter-driven Id1 transgene. We found that Treg cells continued to accumulate as Id1 transgenic mice aged, resulting in a significant increase in Treg cell counts in the thymus as well as in the periphery compared with wild-type controls. Data from mixed bone marrow assays suggest that Id1 acts intrinsically on developing Treg cells. We made a connection between Id1 expression and CD28 costimulatory signaling because Id1 transgene expression facilitated the formation of Treg precursors in CD28−/− mice and the in vitro differentiation of Treg cells on thymic dendritic cells despite the blockade of costimulation by anti-CD80/CD86. Id1 expression also allowed in vitro Treg differentiation without anti-CD28 costimulation, which was at least in part due to enhanced production of IL-2. Notably, with full strength of costimulatory signals, however, Id1 expression caused modest but significant suppression of Treg induction. Finally, we demonstrate that Id1 transgenic mice were less susceptible to the induction of experimental autoimmune encephalomyelitis, thus illustrating the impact of Id1-mediated augmentation of Treg cell levels on cellular immunity.

Objective To investigate the agreement between Rome III and Rome II criteria for diagnosing functional constipation (FC) and to evaluate the accuracy of each constipation symptom for FC diagnosis. Methods Patients with chronic constipation underwent rigorous biochemical and endoscopic/imaging tests to exclude organic and metabolic diseases. The questionnaires including general information, constipation symptoms, and the most troublesome constipation symptoms were completed in a face-to-face survey. The accuracy of constipation symptoms for FC diagnosis was examined using the likelihood ratio. Results Among 184 patients (43 males and 141 females) with chronic constipation, 166 (90.2%) met Rome II criteria and 174 (94.6%) met Rome III criteria for FC, while 166 met both criteria. There was a good diagnostic agreement between the two sets of criteria, with a kappa value of 0.69 and the overall agreement rate was 95.7% (P < 0.001). Based on Rome III criteria, the most accurate symptom for FC diagnosis was sensation of anorectal blockage, followed by straining during defecation and infrequent bowel movements. The most troublesome symptoms reported by patients were lumpy or hard stools, straining during defecation, sensation of incomplete evacuation. More patients indicated that ‘the symptoms in the past 3 months’ was better than ‘those within the past one year’ to reflect their constipation (36.7% vs 6.0%, P < 0.001). Conclusions There is good agreement between Rome III and Rome II criteria for FC diagnosis. Rome III criteria are more practical than Rome II criteria for Chinese patients.

The enthalpy of reaction of the 2-chloroethylideneaminocarbamic acid ethylester formation from chloroacetaldehyde and hydrazinoformic acid ethylester in ethanol at the temperature of 298.15 K has been determined by a microcalorimeter. Experimental results gave the enthalpy of formation Δ r H m θ =(−16.07±0.079) kJ mol −1 , the reaction order of this reaction n=1; the rate constant k=1.190×10−3 s−1 and the free energy of activation Δ G ≠ θ =89.718 kJ mol −1 . The result indicates that the reaction takes place easily at room temperature.

The low temperature heat capacities of ethyl carbazate (C3H8N2O2) were precisely determined with an adiabatic calorimeter over the temperature range from 78 to 371 K. The sample was observed to melt at 318.92 +/- 0.02 K. The molar enthalpy and entropy of the melting as well as the chemical purity of the substance were determined to be 20019 +/- 11 J mol(-1), 62.77 +/- 0.03 J mol(-1) K-l and 99.35 +/- 0.01 mol%, respectively. The extrapolated melting temperature for the absolutely pure compound obtained from fractional melting experiments was 319.181 +/- 0.005 K. (C) 2001 Elsevier Science B.V. All rights reserved.

To the Editor: Japanese encephalitis (JE) epidemics have occurred only in Asia. More than 50,000 cases of JE with ≈10,000 deaths have been reported since 1998 (1,2). The People’s Republic of China reported 5,104 cases and 214 deaths in 2005. Most of these deaths occurred in infants (3,4). During July and August 2006, an outbreak of viral encephalitis occurred in Yuncheng, Shanxi Province, People’s Republic of China. A total of 66 cases (1.32/100,000 population) were reported, including 19 deaths (case-fatality rate 28.8%). The cases had a widespread distribution over 9 counties and involved 37 towns and 61 administrative villages. The ratio of male-to-female patients was 1:0.89. A distinct clinical feature of this outbreak was the age distribution. More than 86% of the patients were >30 years of age, with only 10% of patients 50 years of age (5). We report serologic and virologic findings for the 2006 outbreak of viral encephalitis. Forty-six clinical specimens collected from 34 patients who had a diagnosis of viral encephalitis, including 33 serum samples and 13 cerebrospinal fluid (CSF) samples, were studied. All serum samples were screened for immunoglobulin M (IgM) to West Nile virus (WNV) by using the WNV IgM-capture ELISA kit (PanBio, Brisbane, Queensland, Australia) and for IgM to dengue virus or Japanese encephalitis virus (JEV) by using the JE-Dengue IgM Combination ELISA kit (PanBio). Results for JEV were confirmed by using the JE Virus IgM-Capture ELISA kit (Shanghai B & C Enterprise Development Co. Ltd, Shanghai, People’s Republic of China). WNV-specific or dengue virus–specific IgM was not detected in any samples. JEV-specific IgM was detected in 27 (80%) patients, which indicated recent JEV infections. The other 7 patients were negative for JEV by ELISA and reverse transcription–PCR (RT-PCR). Increases >4-fold in neutralizing antibodies were detected in acute- and convalescent-phase serum samples from 9 patients (10 serum pairs were collected during the outbreak). Attempts were made to detect virus in CSF of patients and in 2,400 mosquitoes. Mosquitoes (mainly Culex spp.) were collected in cow sheds and hog pens around houses and processed into pools of 100. Total RNA was extracted from CSF or mosquito homogenate by using the QIAamp viral RNA extraction kit (QIAGEN, Valencia, CA, USA) according to the manufacturer’s specifications. RT was performed by using Ready-To-Go-You Prime First Strand Beads (Amersham Pharmacia Biotech, Piscatawy, NJ, USA) and a seminested PCR to amplify 492-bp gene fragments of the premembrane (PrM) sequence of JEV by using the Takara LA Taq PCR kit (Takara Bio Inc., Shiga, Japan). The primers were derived from Ishikawa strain genome sequences (GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"AB051292","term_id":"12082323","term_text":"AB051292"}}AB051292). Primers PrMF: 5′-CGT TCT TCA AGT TTA CAG CAT TAG C-3′ (251–275), PrMR1: 5′-CGY TTG GAA TGY CTR GTC CG-3′ (724–743), and PrMR2: 5′-CCY RTG TTY CTG CCA AGC ATC CAM CC-3′ (901–925) were used. JEV PrM gene was amplified from CSF of 6 (46%) of 13 patients and 10 of 24 pools of mosquitoes by using the same seminested RT-PCR. To identify JEV genotype(s) involved in this outbreak, PCR products were sequenced. Eleven sequences (GenBank accession nos. {"type":"entrez-nucleotide-range","attrs":{"text":"EF434264-EF434274","start_term":"EF434264","end_term":"EF434274","start_term_id":"148925198","end_term_id":"148925218"}}EF434264-EF434274) were obtained from 6 patients and 5 pools of mosquitoes. The 11 sequences were compared phylogenetically with17 known JEV strains of the 4 recognized genotypes (classified on the basis of a 240-nt region of the prM gene). As shown in the Figure, the 11 sequences were those of JEV. Figure Phylogenetic analysis of Japanese encephalitis virus strains predicted from premembrane gene sequences. Neighbor-joining tree was generated by using MEGA 3.1 software (www.megasoftware.net) and rooted with Murray Valley encephalitis (MVE) virus sequence ... Further analysis showed that these 11 sequences can be grouped into genotypes I and III. Both genotypes were found in patient and mosquito samples, indicating that these genotypes co-circulated during this JE outbreak. JE has been endemic in Yuncheng for many years (6). A vaccine against JE (SA14–14–2) has been used in this area in infants, but not in adults. This might be 1 reason why a higher adult incidence was found in this outbreak. JEV genotype III had been the predominant genotype in previous years, but genotype I has been recently detected at increased frequencies (7–10). Detection of 2 JEV genotypes in 1 epidemic has not been reported. Whether simultaneous circulation of >1 genotype during an outbreak indicates a new type of emergence of JEV or that this has occurred and not been detected is unknown.

To explore the effects of postprandial diaphragm training (DT) on esophageal acid exposure, esophageal motility and proximal gastric volume at different postprandial periods in patients with gastroesophageal reflux disease (GERD).Thirty GERD patients and 9 healthy subjects (HS) with matched demographic characteristics were enrolled from June 2005 to June 2006 at Peking Union Medical College Hospital. Esophageal manometry with a Dent sleeve catheter and simultaneous esophageal pH monitoring were recorded in a 30-min fasting period and a 120-min postprandial period. The GERD patients were divided into 3 groups: 9 patients received diaphragm training at 1(st) hour after meal (group DT1 h) and another 10 at 2(nd) hour after meal (group DT2 h) whereas no diaphragm training after meal in 11 (group NDT). Ultrasonic imaging of proximal gastric volume was undertaken at 0, 30, 60, 90 and 120 min after meal.(1) The percentage time with pH4 in group DT1 h was lower than that in group NDT in the 120-min postprandial period (0.2% (0-4.1%), 6.6% (2.2%-18.2%), P0.05) and no significant difference of esophageal acid exposure was observed between groups DT2 h and NDT (3.7% (0.1%-17.8%), 6.6% (2.2%-18.2%), P0.05) . (2) Esophagogastric junction (EGJ) and crural diaphragm pressures at the 1(st) hour after meal in group DT1 h were both significantly higher than those in group NDT during diaphragm training ((44.4 ± 8.1) vs(16.2 ± 4.5) mm Hg, (38.2 ± 4.2) vs (9.8 ± 4.5) mm Hg, 1 mm Hg = 0.133 kPa, both P0.05). EGJ and crural diaphragm pressures at the 2(nd) hour after meal in group DT2 h were significantly higher than those in group N-DT during diaphragm training ((53.2 ± 7.5) vs (14.0 ± 3.7) mm Hg, (48.2 ± 6.3) vs (8.9 ± 2.7) mm Hg, both P0.05). There was no change of lower esophageal sphincter pressure (all P0.05). (3) After test meal, the groups DT1 h, DT2 h and N-DT had similar proximal stomach volume (all P0.05).Diaphragm training at the 1(st) hour after meal might reduce the 120-min postprandial esophageal acid exposure in GERD patients. The reduction in esophageal acid exposure may result from enhanced antireflux barrier of EGJ function. Therefore postprandial diaphragm training provides a new approach to conservative treatment of GERD.

The family of inhibitor of differentiation (Id) proteins is a group of evolutionarily conserved molecules, which play important regulatory roles in organisms ranging from Drosophila to humans. Id proteins are small polypeptides harboring a helix-loop-helix (HLH) motif, which are best known to mediate dimerization with other basic HLH proteins, primarily E proteins. Because Id proteins do not possess the basic amino acids adjacent to the HLH motif necessary for DNA binding, Id proteins inhibit the function of E protein homodimers, as well as heterodimers between E proteins and tissue-specific bHLH proteins. However, Id proteins have also been shown to have E protein-independent functions. The Id genes are broadly but differentially expressed in a variety of cell types. Transcription of the Id genes is controlled by transcription factors such as C/EBPβ and Egr as well as by signaling pathways triggered by different stimuli, which include bone morphogenic proteins, cytokines, and ligands of T cell receptors. In general, Id proteins are capable of inhibiting the differentiation of progenitors of different cell types, promoting cell-cycle progression, delaying cellular senescence, and facilitating cell migration. These properties of Id proteins enable them to play significant roles in stem cell maintenance, vasculogenesis, tumorigenesis and metastasis, the development of the immune system, and energy metabolism. In this review, we intend to highlight the current understanding of the function of Id proteins and discuss gaps in our knowledge about the mechanisms whereby Id proteins exert their diverse effects in multiple cellular processes.

The helix-loop-helix transcription factor E2A plays important roles not only in promoting cellular differentiation but also in suppressing cell growth. Id proteins, the inhibitors of E2A, have opposite effects on cell differentiation and growth. To understand the mechanisms by which E2A suppresses cell growth, we examined the role of E2A in regulating the expression of the cyclin-dependent kinase inhibitor p21CIP1/WAF1/SD11, which prevents cell cycle progression upon overexpression. By using transient-cotransfection assays of luciferase reporter constructs in HeLa cells, we have found that overexpression of E2A can transcriptionally activate the p21 gene. To identify the sequences that mediate this activation in the promoter of the p21 gene, we carried out mutational analyses. Out of the eight putative E2A-binding sequences (E1 to E8) in the promoter, the E1 to E3 sequences located close to the transcription start site are found to be essential. In addition, loss of the E boxes in the promoter also reduces p21 expression without cotransfection with E2A in HIT pancreatic cells, where the endogenous E2A-like activity is high. Furthermore, we have also shown that overexpression of E2A in 293T cells activates expression of the endogenous p21 gene at both the levels of mRNA and protein. In correlation with the finding that E47 overexpression leads to growth arrest in NIH 3T3 cells, we have shown that Id1 overexpression in NIH 3T3 cells accelerates cell growth and inhibits p21 expression. Taken together, these results provide insight into the mechanisms by which E2A and Id proteins control cell growth.

The title compound, C9H11N4O2S, consisting of a pyrimidine ring and a thio­urea carboxylic acid methyl ester, possesses mirror symmetry and an intra­molecular N—H⋯N hydrogen bond. In the crystal structure, N—H⋯S hydrogen bonds link symmetry-related mol­ecules to form dimers, which in turn are linked by a C—H⋯O hydogen bond to form a two-dimensional planar structure.

The concept of "three-yin and three-yang" in Shanghan Lun (Treatise on Cold Pathogenic Diseases), a classic written by Zhang Zhongjing in Han Dynasty, has been always the focus of dispute in successive dynasties. The essence of "three-yin and three-yang" has not been fully revealed up till now. Through studying the six divisions of day and night, the six diseases, the combination of syndromes, the complicated diseases, the complete recovery time and the space division of "three-yin and three-yang", the authors draw a conclusion that the "three-yin and three-yang" in Shanghan Lun is a concept of time-sequence, which is associated with the location of disease in space. So it is suggested that the "six diseases" in Shanghan Lun is a categorization for exogenous febrile diseases, and this categorization reveals a sort of inner relationship between the emergence, development, transformation of the febrile diseases and the time.

This present paper describes the development of a reverse-phase high-performance liquid chromatography (RP-HPLC) coupled with UV spectrometry for simultaneous determination of atractylenolide II and III in traditional Chinese medicines and Chinese medicinal preparation. The standard of atractylenolide II and III was isolated from Rhizoma Atractylodes Macrocephalae. From a variety of solvents tested, methanol was found to be the best solvent for simultaneously extracting atractylenolide II and III from Rhizoma Atractylodes Macrocephalae and Chinese medicinal preparation samples. RP-HPLC analysis of the extracts was performed on an analytical column (SHIMADZU ODS, 150 × 4.6 mm2; i.d., 5 μm) equipped with a security guard pre-column system. The results showed that there was a good linearity over the range 0.5–50.0 μg/ml (r > 0.99). The recoveries were more than 95.0 % in Rhizoma Atractylodes Macrocephalae, and the intra- and inter-day coefficients of variation were less than 5.0 % in all cases. The lower limit of quantification (LLOQ) was 0.5 μg/ml. The RP-HPLC method was readily applied to simultaneously quantitate atractylenolide II and III in Rhizoma Atractylodes Macrocephalae and Chinese medicinal preparation (Shenlingbaizhu granule).

Like normal stem cells, cancer stem cells (CSCs) are capable of self-renewal, either by symmetric or asymmetric cell division. They have the exclusive ability to reproduce malignant tumors indefinitely, and to confer resistance in response to radiation or chemotherapy. The ubiquitin modification system plays various roles in physiology and pathology. The key component for the specificity of this system is ubiquitin ligases (E3s). Of these E3s, the majority are RING finger proteins. Many RING finger E3s, such as the Cullin1-Skp1-F-box protein (SCF) E3s, CBL, BRCA1, MDM2 and von Hippel-Lindau tumour suppressor (VHL), are crucial in the regulation of cell-cycle progression and cell differentiation. As a result, many RING finger E3s are implicated in the positive and negative regulation of CSC maintenance. This review summarizes current knowledge in this research field.

To the Editor: Seoul virus (SEOV), a member of the family Bunyaviridae, genus Hantavirus, is primarily carried by Rattus norvegicus rats. Because members of Rattus species are widely distributed, SEOV has the potential to cause human disease worldwide. It has been reported that SEOV causes a milder form of hemorrhagic fever with renal syndrome than Hantaan virus and Dobrava-Belgrade virus and is responsible for 25% of cases of hemorrhagic fever with renal syndrome in Asia (1). Although it is well known that SEOV is endemic to China (2) and South Korea (3), little is known about its distribution in North Korea (4). In September 2009, June and September 2010, and September 2011, a total of 89 R. norvegicus rats were trapped in the city of Hyesan (128°30′E, 41°30′N) during the operation of a cooperative rodent surveillance program of China and North Korea. The captured rodents were euthanized with barbiturate (100 mg/kg), weighed, measured, classified by sex, and then autopsied. Lung samples were probed for the large segment of SEOV by reverse transcription PCR by using the RT primer P14 (5), the primary PCR primers HAN-L-F1 and HAN-L-R1, and the nested PCR primers HAN-L- F2 and HAN-L-R2 (6). PCR products were sequenced by using an ABI 3730 sequencer (Applied Biosystems, Foster City, CA, USA). A high rate of SEOV infection was detected in R. norvegicus rats; 15 (16.8%) of 89 rodent samples tested positive for SEOV by reverse transcription PCR. Infection rates at each surveillance time were 26.7% (4/15) in September 2009, 7.5% (3/40) in June 2010, 28.6% (6/21) in September 2010, and 15.4% (2/13) in September 2011. All infected R. norvegicus rats were adults; 9 were male and 6 were female. The rate of nucleotide substitution in these 15 SEOV amplicons (330 bp; GenBank accession nos. {"type":"entrez-nucleotide-range","attrs":{"text":"KC576788-KC576802","start_term":"KC576788","end_term":"KC576802","start_term_id":"478683850","end_term_id":"478683878"}}KC576788-KC576802, {"type":"entrez-nucleotide","attrs":{"text":"JX853574","term_id":"426273237","term_text":"JX853574"}}JX853574) was calculated by Bayesian Markov chain Monte Carlo analysis using BEAST 1.74 (7). The mean substitution rate, calculated by using the uncorrelated lognormal distribution relaxed molecular clock model and a Bayesian skyline model for the large segment of SEOV, was 8.27 × 10−3 substitutions/site/year, with a 95% high posterior density interval that ranged from 1.02 × 10−4 to 1.79 × 10−2. This substitution rate is about 3 times greater than that for middle and small segments (2). Phylogenetic relationships were assessed by using the uncorrelated lognormal distribution relaxed molecular clock model with the SRD06 substitution model (8) in BEAST 1.74. The Hantaan virus strain AA57 (GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"AB620033","term_id":"326784691","term_text":"AB620033"}}AB620033) sequence was used as the outgroup. The resulting phylogenetic tree (Figure) showed that SEOV strains in the city of Hyesan shared >97.3% identity and were all clustered in their own lineages, subdivided into 2 co-existing sublineages. Although the geographic distance from Hyesan to northeastern China (e.g., Liaoning Province) is much less than that between northeastern and southeastern China (e.g., Zhejiang Province) or central China (e.g., Hubei Province), the phylogenetic distance between SEOV strains in North Korea and those in each location in China in clade A, calculated by using MEGA5.1 (9), was 0.03, but was only 0.01–0.02 between locations in China. Figure Phylogenetic tree, based on a 330-bp amplicon of the Seoul virus (SEOV) RNA-dependent RNA polymerase gene, depicted in FigTree1.4.0 (http://www.molecularevolution.org/software/phylogenetics/figtree). The tree was generated by using the uncorrelated lognormal ... One possible explanation for this discrepancy in phylogenetic and geographic distances between SEOV strains in China and those in North Korea may be differences in the extent of human contact. Although human interactions among different regions of China are extensive, by comparison, those between China and North Korea are considerably reduced for political reasons. In addition, combining with small segment (GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"HQ992815","term_id":"341842442","term_text":"HQ992815"}}HQ992815) sequence analysis (data not shown), the fact that SEOV strain L0199 from Laos were not clustered in clade A-D(2) showed that Laos was another possible area of origin for SEOV. Our work contributes to the known epidemiology of exposure to the SEOV pathogen in Hyesan. Hyesan adjoins Changbai County in Jilin Province of China. However, SEOV was not detected in Changbai County during the surveillance program (data not shown), which was consistent with previous research (10). This study further highlights the need for long-term surveillance.

To evaluate the status of complementary feeding frequency (CFF) for infants and young children in southwestern China.A total of 3644 infants and young children aged 6 - 24 months from urban and rural areas of Chengdu, Kunming and Guiyang were selected by stratified random cluster sampling from March to June in 2011. Data of CFF in the recent one month were collected through the questionnaires, and assessed quantitatively by a new comprehensive evaluation system. Level and distribution characteristics of CFF for infants and young children among different month groups in urban and rural areas were analyzed.Average CFF score was 8.1 ± 3.1, and the score for all was 54.1% of total score (15 points). The average score of urban and rural groups was 8.9 ± 3.0 (59.3% of total score) and 7.4 ± 3.0 (49.1% of total score) respectively (t = 15.60, P0.05). Ratio of ≥ 80.0% of total CFF score was 12.2% (443/3644) for all. The rate of urban and rural group was 18.0% (324/1796) and 6.4% (119/1848) respectively (χ(2) = 136.64, P0.05). Average CFF score in 6 - 8, 9 - 11 and 12 - 24 months groups was 7.0 ± 2.9 (46.4% of total score), 8.1 ± 3.0 (54.1% of total score) and 9.0 ± 3.0 (60.1% of total score) respectively (F = 148.27, P0.05). The CFF score increased with months growing.Status of CFF for infants and young children in southwestern China is generally inadequate, with differences between urban and rural groups. This problem is more serious in early month infants and rural areas and should be emphasized and improved.

Xiaocheng Gu, a crystallographer and professor emeritus, in the College of Life Sciences, Peking University, died on November 3, 2012, from complications of kidney cancer. Professor Gu was born on February 13th, 1930 in Shanghai, China. Professor Gu attended Peking University, the Department of Plant Sciences, for her bachelor's degree (1948-1952). She obtained advanced training in protein crystallography in the Department of Chemistry, Peking University (1961-1964). Like most of the intellectuals in China during that period, Professor Gu's scientific career was interrupted by the “Cultural Revolution” from 1966 through 1976. From 1978 to 1984, Professor Gu was the Deputy Chairperson, Department of Biology, Peking University. In 1985, she became the Chairperson of the same department, a position that she served until 1992. In 1995, she became Professor Emeritus.

Hematopoietic stem cells (HSCs) maintain life-long blood supply but are inevitably exposed to various inflammatory stimuli, which have been shown to be harmful for HSC integrity but the mediators of the deleterious effects have not been fully identified. Here, we show that daily injection of mice with 1 µg of LPS for 30 days triggers a storm of inflammatory cytokines. LPS injection also stimulated the transcription of the Id1 gene in HSCs in vivo but not in vitro, suggesting an indirect effect. To determine the effects of LPS treatment on HSC function and to evaluate the significance of Id1 expression, we assess the repopulating potential of wild type and Id1 deficient mice, which were subjected to a 30 day regimen of LPS treatment. We found that LPS caused dramatic reduction in the long-term but not short-term repopulating activity of wild type but not Id1 deficient HSC. This treatment also led to increases in HSC counts, decreases in BrdU-label retention and disturbance of quiescence detected by Ki67 staining in wild type but not Id1 deficient mice. Together, it appears that Id1, at least in part, plays a role in LPS-induced damage of HSC integrity.

The helix-loop-helix (HLH) Id proteins have been reported to function as inhibitors of various differentiation programs. The HLH motif mediates dimer formation between Id and the basic HLH transcription factors. Since Id proteins lack the basic region responsible for DNA binding, the heterodimers cannot bind to DNA. Id proteins have also been found to be involved in early B-cell differentiation. They are expressed at high levels in progenitor B cells (pro-B cells), and the expression is diminished in pre-B cells and mature B cells. This expression pattern correlates inversely with basic HLH protein activity and immunoglobulin enhancer function in B-cell development. Regulation of Id expression may play an important role in transcriptional control of immunoglobulin genes and therefore in B-cell differentiation. We have characterized the regulatory elements of the Id1 gene. Using stable transfectants, transient transfection, and mobility shift assays, we have identified an 8-bp element designated PBE (pro-B enhancer) downstream of the Id1 gene that is responsible for a pro-B-cell-specific enhancer activity. A pro-B-cell-specific protein complex was found to bind to the 8-bp PBE element. Substitution mutagenesis at this binding site showed that it is indeed of functional importance in regulating the pro-B-cell-specific expression of the Id1 gene.

In this study, PCR-DGGE was used to analyze the microbial community structure in lactic acid fermentation from kitchen waste. The results showed that with Lactobacillus amylophilus inoculation, both the microbial diversity and lactic acid production in the open fermentation system were higher than those in the sterilized fermentation system. These results indicated that the microbial diversity and the lactic acid production have great correlation in the kitchen waste fermentation system. Through analyzing the sequence of some DNA bands excised from the DGGE gel, it showed that in addition to the inoculation of Lactobacillus amylophilus there were some indigenous lactic acid bacteria, such as Lactobacillus sp., Lactobacillus casei, Lactobacillus plantarum and indigenous hydrolytic bacteria, such as Pseudomonas sp.. These indigenous bacteria can help to promote lactic acid production. PCR-DGGE is feasible for analyzing the dynamic changes of microbial community structure in kitchen waste with complicated composition.

Microwave-alkali and steam-alkali coupled pretreatments were carried out to improve the yield and optical purity of l-lactic acid produced using vinasse fermentation. Lactobacillus casei was inoculated into the system to initiate fermentation. Polymerase chain reaction denaturing gradient gel electrophoresis was used to analyze the microbial community during fermentation with and without the pretreatments. The original bacterial genus in vinasse was essentially inactivated, whereas L. casei became the dominant genus after 24h of fermentation. The system subjected to microwave-alkali coupled pretreatment released more reducing sugars and produced more lactic acid (up to 30.32g/L), which is twice that without pretreatment. In addition, the proportion of lactic acid in the organic acids also increased. The optical purity of the l-lactic acid produced under the microwave-alkali coupled pretreatment reached 91%, which is 2% higher than that under the steam-alkali coupled pretreatment and 7% higher than that under the control conditions. Therefore, the microwave-alkali coupled pretreatment is an effective method for the highly efficient bioconversion of vinasse into bioenergy.

Functional dyspepsia-epigastric pain syndrome (FD-EPS) is characterized pathophysiologically by visceral hypersensitivity, but the effect of the temperature stimulation on gastric function has been seldom studied. The purpose of this study was to investigate the effects of liquid nutrients at different temperatures on the gastric accommodation, sensitivity, and gastric-wall compliance of healthy subjects (HS) and FD-EPS patients. Ten FD-EPS patients (Roma III criteria) and ten HS were recruited into the study. Intragastric pressure (IGP) and gastric perfusion were measured and compared following the administration of liquid nutrients at 37 °C on day 1 and at 8 °C on day 2. Seven patients developed abdominal discomfort or abdominal pain after being given cold liquid nutrient. The administration of liquid nutrient at 8 °C resulted in an increase of IGP in HS (P = 0.044), a significant decrease in gastric perfusion (P

The ld proteins are inhibitors of the basic-helix-loop-helix (bHLH) transcription factors. In the B cell lineage, the Id1 and Id2 genes are expressed in pro-B cells and down-regulated during differentiation. To determine the role of bHLH proteins and the significance of down-regulation of Id genes in B cell development, transgenic mice constitutively expressing the Id1 gene were generated. Their phenotype suggests that B cell development is impaired at an early stage. Primarily, these mice have few B220 + IgM + mature B or B220 + CD43 − pre-B cells in the bone marrow, reduced frequencies of V(D)J and V K J K recombination of the immunoglobulin loci, and lower expression levels of the immunoglobulin, RAG-1, RAG-2 , and λ5 genes.

Janus kinase (JAK)-signal transducer and activators of transcription (STAT) signaling pathways play crucial roles in lymphopoiesis. In particular, JAK3 has unique functions in the lymphoid system such that JAK3 ablation results in phenotypes resembling severe combined immunodeficiency syndrome. This review focuses on the biochemistry, immunological functions, and clinical significance of JAK3. Compared with other members of the JAK family, the biochemical properties of JAK3 are relatively less well characterized and thus largely inferred from studies of JAK2. Furthermore, new findings concerning the cross-talks between Notch and JAK signaling pathways through ubiquitin-mediated protein degradation are discussed in more detail.

T cell lymphoblastic leukemia (T-ALL) is known to be associated with chromosomal abnormalities that lead to aberrant expression of a number of transcription factors such as TAL1, which dimerizes with basic helix-loop-helix (bHLH) E proteins and inhibits their function. Activated Notch receptors also efficiently induce T cell leukemogenesis in mouse models. Interestingly, gain-of-function mutations or cryptic transcription initiation of the Notch1 gene have been frequently found in both human and mouse T-ALL. However, the correlations between these alterations and overall Notch activities or leukemogenesis have not been thoroughly evaluated. Therefore, we made use of our collection of T cell lymphomas developed in transgenic mice expressing Id1, which like TAL1, inhibits E protein function. By comparing expression levels of Notch target genes in Id1-expressing tumors to those in tumors induced by a constitutively active form of Notch1, N1C, we were able to assess the overall activities of Notch pathways and conclude that the majority of Id1-expressing tumors had elevated Notch function to a varying degree. However, 26% of the Id1-expressing tumors had no evidence of enhanced Notch activation, but that did not delay the onset of tumorigenesis. Furthermore, we examined the genetic or epigenetic alterations thought to contribute to ligand-independent activation or protein stabilization of Notch1 and found that some of the Id1-expressing tumors acquired these changes, but they are not uniformly associated with elevated Notch activities in Id1 tumor samples. In contrast, N1C-expressing tumors do not harbor any PEST domain mutations nor exhibit intragenic transcription initiation. Taken together, it appears that Notch activation provides Id1-expressing tumor cells with selective advantages in growth and survival. However, this may not be absolutely essential for lymphomagenesis in Id1 transgenic mice and additional factors could also cooperate with Id1 to induce T cell lymphoma. Therefore, a broad approach is necessary in designing T-ALL therapy.

Japanese encephalitis virus (JEV), a mosquito-borne zoonotic pathogen, is one of the major causes of viral encephalitis worldwide. Previous phylogenetic studies based on the envelope protein indicated that there are four genotypes, and surveillance data suggest that genotype I is gradually replacing genotype III as the dominant strain. Here we report an evolutionary analysis based on 98 full-length genome sequences of JEV, including 67 new samples isolated from humans, pigs, mosquitoes, midges. and bats in affected areas. To investigate the relationships between the genotypes and the significance of genotype I in recent epidemics, we estimated evolutionary rates, ages of common ancestors, and population demographics. Our results indicate that the genotypes diverged in the order IV, III, II, and I and that the genetic diversity of genotype III has decreased rapidly while that of genotype I has increased gradually, consistent with its emergence as the dominant genotype.

Economic development and increased tourism in the southern region of Yunnan Province in China, adjacent to several countries in Southeast Asia, has increased the likelihood of import and export of vectors and vector-borne diseases. We report the results of surveillance of mosquitoes and mosquito-borne arboviruses along the border of China-Myanmar-Laos in 2005 and 2006, and information associating several arboviruses with infections and possibly disease in local human populations. Seventeen mosquito species representing four genera were obtained, and 14 strains of mosquito-borne viruses representing six viruses in five genera were isolated from Culex tritaeniorhynchus. In addition, IgM against Japanese encephalitis virus, Sindbis virus, Yunnan orbivirus and novel Banna virus was detected in acute-phase serum samples obtained from hospitalized patients with fever and encephalitis near the areas where the viruses were isolated. This investigation suggests that Japanese encephalitis virus, Sindbis virus, and lesser-known arboviruses circulate and may be infecting humans in the China-Myanmar-Laos border region.

Alzheimer’s disease (AD) is a neurodegenerative disease characterized by extracellular amyloid beta (Aβ) deposition and intracellular neurofibrillary tangle formation. Monocyte is part of the innate immune system and can effectively remove dead cells and debris. It has been suggested that Aβ can recruit monocytes into brain in AD mice, resulting in restriction of cerebral amyloidosis. However, monocyte may act as a double-edged sword, either beneficial (e.g., clearance of Aβ) or detrimental (e.g., secretion of neurotoxic factors). In addition, recent studies indicate that in AD patients, Aβ phagocytosis by monocytes is ineffective. The present review mainly summarized the current knowledge on monocytes and their potential roles in AD.

Through observing the morphological changes of the prepared influenza viruses (H1N1) treated with the different concentration of Nonidet P-40 solutions and added with antibody against the influenza virus using transmission electron microscopy (TEM), to explore the principle of application of the internal antibody for immunoassay of influenza virus.Through treating the virus samples with serial diluted Nonidet P-40 solutions from 0.01% to 0.2% and/or not adding antibody against the influenza virus, then investigating the samples by TEM to obtain the morphological changes of the virions.The serial images show that the denudation degree of the virions is proportional with the rise of NP-40 concentration, and partly denuded virion image appeared at 0.1% NP-40 treatment, also under this condition no influence was observed on antibody binding to the virus.This work demonstrated the interaction between influenza virion and its antibody under nonionic surfactants existing, which supports the advantages of sample preparation for immunoassay enveloped virus using internal antibody theoretically.

Basic helix–loop–helix E proteins play critical roles in B-cell development by stimulating B cell-specific gene expression and immunoglobulin gene rearrangement. The function of E proteins can be effectively suppressed by their naturally occurring inhibitors, Id1 to 4. Ectopic expression of Id1 has been shown to block B-cell development at the early pro-B cell stage. However, whether Id1 plays a physiological role in controlling B lymphopoiesis was not known. Although Id1-deficient mice do not exhibit significant abnormalities in steady-state B lymphopoiesis, we detected more robust B-cell engraftment in transplant recipients of Id1-deficient bone marrow compared to those of wild-type donor cells. In culture, Id1 ablation dramatically enhances B-lineage cell production without any marked effects on myeloid differentiation. Consistently, Id1 expression was found in pro-B but not pre-B cells as measured by enhanced green fluorescent protein (EGFP) fluorescence and by quantitative reverse transcription-PCR. Although loss of Id1 did not alter the number of B-cell colonies generated from whole bone marrow or the proliferation rate of developing B cells, B-cell colonies were detectable at a much earlier time point and the size of the colonies were larger. Therefore, we infer that Id1-deficient progenitors possess higher potential to differentiate to the pre-B cell stage when a proliferative burst occurs. Taken together, we present evidence to suggest that Id1 plays a physiological role in restraining the developmental progression, which may be important for proper B-cell differentiation in the bone marrow.

To isolate and identify Banna virus (BAV) from mosquitoes collected in Mengla county of Yunnan province.Mosquito samples were collected in houses and stock yards in Mengla county, 2008. Mosquitoes were homogenized and incubated onto both C6/36 and BHK21 cells. The new isolate was identified by using ELISA and RT-PCR. The sequences of segment 5, 8 and 11 of BAV were amplified by RT-PCR and determined. Phylogenetic analysis on the new BAV were performed using MEGA4 program.1731 mosquitoes representing 7 species were collected with one strain of BAV isolated and identified. Phylogenetic analysis based on sequences of segment 8 showed the new isolate was closed to BAV strain isolated in Yunnan, but segment 11 sequence was closed to Vietnam strain.Results of phylogenetic analysis implied that the BAV re-assortment might have been occurred both in Chinese and Vietnam strains.

The interaction between 4-(4-fluorobenzylideneamino)-5-propyl-4H-1,2,4-triazole-3-thiol (FBTZ) and human serum albumin (HSA) under simulative physiological conditions was investigated by fluorescence, UV-vis absorption and circular dichroism (CD) spectroscopy as well as molecular modeling method. Fluorescence spectroscopic data showed that the fluorescence quenching of HSA was a result of the formation of FBTZ-HSA complex. According to the modified Stern-Volmer equation, the effective quenching constants (K (a)) of FBTZ to HSA were obtained at three different temperatures. The enthalpy change (ΔH) and entropy change (ΔS) were calculated on the basis of van't Hoff equation, and the results showed that hydrogen-bonding and van der Waals forces were the dominant intermolecular forces to stabilize the complex. Site marker competitive replacement experiments demonstrated that the binding of FBTZ to HSA primarily took place in sub-domain IIA (Sudlow's site I). The binding distance (r) between FBTZ and the tryptophan residue of HSA was estimated according to the theory of fluorescence resonance energy transfer. The conformational investigation showed that the presence of FBTZ induced some changes of secondary structure of HSA. Molecular modeling study further confirmed the binding mode obtained by experimental study.

To develop a fast, high-throughput screening method with suspension array technique for simultaneous detection of biothreat bacteria.16 S rDNA universal primers for Bacillus anthracis, Francisella tularensis, Yersinia pestis, Brucella spp.and Burkholderia pseudomallei were selected to amplify corresponding regions and the genus-specific or species-specific probes were designed. After amplification of chromosomal DNA by 16 S rDNA primers 341A and 519B, the PCR products were detected by suspension array technique. The sensitivity, specificity, reproducibility and detection power were also analyzed.After PCR amplification by 16 S rDNA primers and specific probe hybridization, the target microorganisms could be identified at genus level, cross reaction was recognized in the same genus. The detection sensitivity of the assay was 1.5 pg/microl (Burkholderia pseudomallei), 20 pg/microl (Brucella spp.), 7 pg/microl (Bacillus anthracis), 0.1 pg/microl (Francisella tularensis), and 1.1 pg/microl (Yersinia pestis), respectively. The coefficient of variation for 15 test of different probes was ranged from 5.18% to 17.88%, it showed good reproducibility. The assay could correctly identify Bacillus anthracis and Yersinia pestis strains in simulated white powder samples.The suspension array technique could be served as an opening screening method for biothreat bacteria rapid detection.

A rapid ultra-performance liquid chromatography-electrospray ionization tandem mass spectrometric (UPLC-ESI-MS/MS) method is developed for the qualitative identification of constituents in the flower buds of seven Lonicera species. The optimal condition of separation and detection were achieved on an AcQuity UPLC BEH C18 column with a gradient elution with acetonitrile and 0.1% acetic acid within 17 min. Among the 33 constituents detected, 6 caffeoylquinic acids (including caffeic acid), 8 flavonoids and 8 iridoid glycosides were characterized based on their fragmentation patterns in collision-induced dissociation (CID) experiments and/or by comparison with standard compounds. In addition, to statistically establish the correlation and discrimination of the Lonicera species, principle component analysis (PCA) was applied in this study. Lonicera samples were divided into well-defined groups directly related to their species based on PCA in terms of the log transformed relative contents of the major caffeoylquinic acids (including caffeic acid) as the variables. All of results indicated that the method presented here is able to classify the sample species and to reveal characteristic details of the chemical constituents of different Lonicera species.

To analyze the molecular characteristics of Japanese encephalitis virus (JEV) isolated in Northwestern Yunnan province, and to clarify the differences between the strains isolated in Northwestern and other parts of Yunnan province.PrM, E and 3' untranslated region nucleotide acid sequences of the isolates were amplified by RT-PCR and then sequenced. Sequence alignment and phylogenetic analysis were performed by using Clustal 1.8X, DNASTAR, GENEDOC and Mega 3.1 programs.12 of the 13 isolates of JEV obtained in Northwestern Yunnan were identified as genotype I, only one strain was genotype III of JEV. The 12 strains of genotype I were clustered in different branches with other isolates obtained in other parts of Yunnan province. Data from sequence analysis on E gene found that the nucleotide identity was 0.2%-13.9% between the Northwestern isolates and other Yunnan strains. There were two kinds of nucleotides deletion patterns at 3' untranslated region with three and one deletions was found after termination codon in genotype I and III isolates, respectively.There were two genotypes of I and III in 13 strains of JEV in this study and genotype I isolates were predominant (12/13). There were no apparent differences in E gene sequence between isolates obtained in the Northwestern and other parts of Yunnan. Three deletions were found in 3' untranslated region in genotypes I isolates and one deletion was in genotypes III.

The 4,5-dihydro-1,2,4-triazole Schiff base derivatives were synthesized with 4-amino-4,5-dihydro-3-(phenoxy- methyl)-1H-1,2,4-triazole-5-thione and substituted benzaldehydes. The chemical structures of the compounds have been confirmed by 1H NMR, IR spectra and elemental analysis. Their biological activities were investigated, and the results showed that they exhibited good fungicidal activities.

Eight Schiff bases of 3-amino-1H-1,2,4-triazole have been synthesized by glacial acetic acid catalyzed condensation of 3-amino-1H-1,2,4-triazole with substituted benzaldehyde and structurally confirmed by IR, 1H NMR and elemental analysis. The preliminary bioassay showed that the title compounds exhibited good fungicidal activities.

Technology standards are the commanding heights of the competition of current high and new technology industry. Behind of this competition is enormous commercial profit. Starting from the network effect property of the technology standard product, by structuring the utility equation of the technology standard product and copying dynamic equation of the evolutionary game, the competition mechanisms of technology standard is analyzed under circumstances of consumer preferences and without consumer preferences respectively, and the factors are also analyzed that can influence the success of technology standard competition combined with the existing cases, finally the relevant countermeasures are proposed.