Objective:The present study aims at setting up an evaluation index system aiming at providing a set of measurement tools for population & health informatization developing level for administrative departments. Meth-ods: Delphi method, analytic hierarchy process, Synthetic Index Method and some mathematical or statistical meth-ods were used respectively to select the primary indexes, calculate the weight and combined weight of each index, build up the synthetic evaluation mode,and test the evaluation system. Results:According to the results of this work, the evaluation index system has been established which is composed of 6 level 1 indexes, 16 level 2 indexes, and 33 level 3 indexes, and it covers 6 aspects, such as information resources construction, information infrastructure con-struction, information technology application, informatization talents and the information policy, planning, and stand-ards. The weights of these indexes were also calculated. The synthetic evaluation model has been built. Finally, the reliability, the validity and the discrimination of the evaluation system were tested. Conclusions:The evaluation index system has been established, which is able to be used in the evaluation, and to some extent guide the work of popula-tion & health informatization construction.

This article aimed to study the antigenicity of nucleocapsid proteins (NPs) in six pathogenic phleboviruses and to provide theoretical evidence for the development of serological diagnostic reagents. NPs of six pathogenic phleboviruses were expressed and purified using a prokaryotic expression system and rabbits were immunized with individual recombinant NPs. Cross-reactions among NPs and rabbit sera were determined by both indirect ELISA and Western blotting analyses, and the sera titer was determined by indirect ELISA. Furthermore, sera from SFTS patients were also detected by each recombinant NP as a coating antigen using indirect ELISA. The cross-reactions and the sera titer were subsequently determined. Both the concentration and purity of recombinant NPs of six pathogenic phleboviruses met the standards for immunization and detection. The results of indirect ELISA and Western blotting showed that each anti-phlebovirus NP rabbit immune serum had potential serological cross-reactivity with the other five virus NP antigens. Furthermore, the sera from SFTS patients also had cross-reactivity with the other five NP antigens to a certain extent. Our preliminary study evaluated the antigenicity and immune reactivity of six pathogenic phleboviruses NPs and laid the foundation for the development of diagnostic reagents.
Animals ; Antibodies, Viral ; immunology ; Antigens, Viral ; genetics ; immunology ; Cross Reactions ; Humans ; Nucleocapsid Proteins ; genetics ; immunology ; Phlebotomus Fever ; diagnosis ; immunology ; virology ; Phlebovirus ; classification ; genetics ; immunology ; isolation & purification ; Rabbits

Objective To explore the application value of fiberoptic bronchoscopy in the diagnosis and treatment for refractoriness pneumonia in children.Methods Sixty-two cases of refractoriness pneumonia who had not been cured with normal anti-infective therapy for 2 weeks in Guiyang Children′s Hospital from Jun.2005 to Jun.2009 were selected.Fiberopic bronchoscopy Olympus BF-3C30 or BF-P40 examination was applied after routine local infiltration anesthesia to diagnose and local treat refractoriness pneumonia in 62 children.Results There were 27 cases(43%) of simple inflammation,20 cases(32%) of foreign bodies with concurrent infection,7 cases(11%) of dndobronchial tuberculosis,4 cases(6%) of dysplasia with concurrent infection,2 cases(3%) of bronchiectasis,and 2 cases(3%) of pulmonary hemosiderosis,among them,dysplasia were diagnosed on children under 1 year old,foreign bodies with infection on 1-3 years old,and tuberculosis and bronchiectasis on school age.Eleven cases(40%) of simple inflammation were identified various pathogen by bronchoalveolar la-vage fluid culture,mycoplasma PCR test and bronchial brushing and microscopic examination.Forty-seven cases of bronchi obstruction and stenosis caused by inflammation were irrigated and treated by drug and achieved satisfactory,among them,15 cases(79%) of pulmonary atelectasis were recovered with the treatment of fiberoptic bronchoscopy within 2 weeks,14 cases(70%) of foreign bodies were succeed in taking choked thing from bronchus with fiberoptic bronchoscopy.Conclusion The fiberoptic bronchoscopy play an important role in the etiological diagnosis and therapy of refractoriness pneumonia in children.

To establish a MacELISA method for the detection of IgM antibodies against Chikungunya virus (CHIKV), we prepared virus like particle (VLP) antigens of CHIKV using the whole structural protein C-E3-E2-6K-E1 encoding gene with a baculovirus expression system in Sf9 insect cells. The VLPs were purified and used to immunize Kunming mice. Then, polyclonal antibodies were purified from the samples of ascites with a protein G HiTrap SP column and labeled with horseradish peroxidase. A MacELISA method for the detection of IgM antibodies against CHIKV was assembled with goat anti-human IgM antibody, VLP antigens and an enzyme-labeled polyclonal antibody. The results were evaluated with a serum panel containing serum samples from laboratory-confirmed CHIK, HFRS patients, healthy donors, and commercially available CHIKV IgM as a quality control. It was shown that the MacELISA had a specificity of 99% (99/100), the coefficients of variation (CoV) within a plate were <10%, and the CoV of different ELISA plates in terms of the plate variation coefficient was <15%. A comparative analysis was performed to compare the current method against a commercial CHIKV IgM antibody detection kit for IIFA-IgM. The detection limit of MacELISA was significantly lower than that of the IIFA-IgM commercial kit (P< 0.0001). Here, we demonstrate that the VLP-based MacELISA is a promising tool for the early diagnosis and epidemiological investigation of CHIKV infection, with a high level of sensitivity and specificity for the detection of IgM antibodies against CHIKV.
Animals ; Antibodies, Viral ; blood ; Chikungunya Fever ; blood ; diagnosis ; virology ; Chikungunya virus ; immunology ; isolation & purification ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Immunoglobulin M ; blood ; Mice

This study aims to investigate whether the nucleoprotein (NP) of severe fever with thrombocytopenia syndrome virus (SFTSV) can impact the cellular immunity of host cells. Gene segments that encode the NP and non-structural protein (NSs) of SFTSV were inserted into eukaryotic expression vector VR1012. Host proteins that interact with NP and affect immunity were identified with co-immunoprecipitation (IP), SDS-PAGE, mass spectrometry (MS), and Western blot. Co-localization of NP and the identified host proteins was confirmed by confocal microscopy. A 60kD SSA/Ro, a protein related to immunity, interacted with NP, as found by IP and MS. Confocal microscopy showed that NP and SSA/Ro were co-localized in cytoplasm. These results indicated that SFTSV NP may specifically bind to 60kD SSA/Ro and cause a series of immune responses and clinical symptoms.
Bunyaviridae Infections ; genetics ; metabolism ; virology ; HEK293 Cells ; Humans ; Nucleoproteins ; genetics ; metabolism ; Phlebovirus ; genetics ; metabolism ; Protein Binding ; Ribonucleoproteins ; genetics ; metabolism ; Viral Proteins ; genetics ; metabolism

BACKGROUNDThe mechanisms of action for volatile anesthetics remain unknown for centuries partly owing to the insufficient or ineffective research models. We designed this study to develop three strains derived from a wild-type Drosophila melanogaster with different sensitivities to volatile anesthetics, which may ultimately facilitate molecular and genetic studies of the mechanism involved.

METHODSMedian effective doses (ED(50)) of sevoflurane in seven-day-old virgin female and male wild-type Drosophila melanogaster were determined. The sensitive males and females of percentile 6 - 10 were cultured for breeding sensitive offspring (S(1)). So did median ones of percentile 48 - 52 for breeding median offspring (M(1)), resistant ones of percentile 91 - 95 for breeding resistant offspring (R(1)). Process was repeated through 31 generations, in the 37th generation, S(37), M(37) and R(37) were used to determine ED(50) for enflurane, isoflurane, sevoflurane, desflurane, halothane, methoxyflurane, chloroform and trichloroethylene, then ED(50) values were correlated with minimum alveolar concentration (MAC) values in human.

RESULTSFrom a wild-type Drosophila melanogaster we were able to breed three strains with high, median and low sevoflurane requirements. The ratio of sevoflurane requirements of three strains were 1.20:1.00:0.53 for females and 1.22:1.00:0.72 for males. Strains sensitive, median and resistant to sevoflurane were also sensitive, median and resistant to other volatile anesthetics. For eight anesthetics, ED(50) values in three strains correlated directly with MAC values in human.

CONCLUSIONSThree Drosophila melanogaster strains with high, median and low sensitivity to volatile anesthetics, but with same hereditary background were developed. The ED(50) are directly correlated with MAC in human for eight volatile anesthetics.

Anesthetics, Inhalation ; pharmacology ; Animals ; Chloroform ; pharmacology ; Drosophila melanogaster ; drug effects ; growth & development ; Enflurane ; pharmacology ; Female ; Halothane ; pharmacology ; Isoflurane ; analogs & derivatives ; pharmacology ; Male ; Methoxyflurane ; pharmacology ; Methyl Ethers ; pharmacology ; Trichloroethylene ; pharmacology

Objective To evaluate the quality of Radix Saposhnikoviae ( Fangfeng ) by using grey relational analysis. Methods The content of six characteristic components of alcohol extract, total polysaccharides, prim-O-glucosylcimifugin, cimifugin, 4′-O-beta-glucopyranosyl-5-O-methylvisamminol and sec-O-glucosylhamaudo were determined in altogether 20 Fangfeng samples. Based on grey relational analysis, the model of quality evaluation was established, by measuring the relative correlation defined. Results The results of quality evaluation were in accordance with the market quality grade of these 20 Fangfeng samples. Conclusion The study is expected to provide a novel way to classify the quality grade and evaluate the quality of Fangfeng in market.

Objective To investigate the dynamic changes of cerebral blood flow defined bytranscranial color Doppler (TCD) and their correlation with the severity and prognosis of acute ischemicstroke. Methods Fifty-six consecutive patients with the primary onset of acute ischemic strokereceived dynamic examinations with TCD upon admission and on days 2, 4, 7, 14 and 28 following theonset, and the mean blood flow velocity and pulsatility index were recorded. Results TCDexaminations upon admission revealed high abnormal rate. The mean blood flow velocity of the middlecerebral artery (VmMCA) on the lesion side was significantly lower than that on the contralateral side andthat of the control group. The blood flow velocity of the anterior cerebral artery (VmACA) on the lesion sidewas significantly increased compared with that on the contralateral side and that of the control group.VmMCA of main stem occlusion group {23.45 [7.55,38.72] cm/s}, central branch infarction group {52.53[45.56,66.93] cm/s}, and cortical branch of cerebral arteries infarction group {38.44[31.67,52.45] cm/s}was significantly lower than that of the control group {66.51 [56.64,71.43] cm/s}. VmMCA graduallyincreased in these patients after the treatment, showing significant difference from that measured uponadmission. The PI of the middle cerebral artery increased after admission, with significant difference fromthat recorded on admission and that of the control group. Conclusion TCD has high diagnostic valuefor ischernic stroke and may help evaluate the severity of the condition and the prognosis of the patients.

OBJECTIVETo investigate the effects of sub-micro emulsion composition on cellular uptake and disposition of incorporated lipophilic drug.

METHODSSub-micro emulsions containing 10 % oil, 1.2 % lecithin and 2.25 % glycerol were prepared, and the fluorescent agent coumarin 6 was used as a model drug. The effects of oil types, co-surfactants and cationic lipid on uptake and elimination kinetics of 6-coumarin in HeLa cells were studied. The uptake mechanism of sub-micro emulsions was further investigated.

RESULTSOil type and Tweens had no influence on the cellular uptake. Modifications of surfactants with Span series increased the cellular influx, among which Span 20 with hydrophilic-lipophilic balance (HLB) value of 8.6 was the best enhancer. The intracellular drug level reached up to (46.09 ± 1.98)ng/μg protein which had significant difference with control group [(38.54 ± 0.34)ng/μg protein]. The positively charged emulsions significantly increased the uptake rate constant and elimination rate constant which were 4 times and 1.5 times of those in anionic groups, respectively. The uptake enhancement was also observed in cationic emulsions, cellular concentrations at plateau were (42.73 ± 0.84)ng/μg protein, which was about 3 times of that in anionic emulsions [(15.71 ± 0.74)ng/μg protein], when extracellular drug concentration kept at 100 ng/ml. Cationic emulsions delivered the payload mainly by direct drug transfer to contacted cells, while the negative ones depended on both drug passive diffusion and clathrin-mediated endocytosis of drug containing oil droplets which accounted for 20% of the intracellular drug.

CONCLUSIONInterfacial characteristic of sub-micro emulsions such as co-surfactants HLB as well as zeta potentials can influence lipophilic drug both in cellular uptake and elimination.

Anions ; Cations ; Coumarins ; pharmacokinetics ; Emulsions ; Endocytosis ; HeLa Cells ; Humans ; Surface-Active Agents ; pharmacokinetics ; Thiazoles ; pharmacokinetics

OBJECTIVETo obtain the functional information of AY358935 gene.

METHODSThe properties, subcellular location, and structure of AY358935 protein, and the expression profile of AY358935 gene were analyzed by bioinformatics software and the biological functions of the gene were predicted. AY358935 expression was detected by Western blot analysis in early virus infection.

RESULTSAY358935 was evolutionally conserved. The human AY358935 protein had an amino acid similarity of 74%, 60%, 38% and 33% with its counterpart in horses, mice, zebrafish and Xenopus laevis, respectively. Bioinformatics analysis indicated that AY358935 protein was located likely in the mitochondria. There was a N-terminal signal peptide and single transmembrane structure in AY358935 protein, which contained several phosphorylation sites. The secondary structure mainly comprised of alpha helices and random coils. AY358935 was ubiquitously expressed in normal tissues and carcinomas and regulated by the expression of double-stranded RNA-dependent protein kinase. AY358935 protein expression was obviously upregulated in cells 2 h after infection by vesicular stomatitis virus.

CONCLUSIONAs a predicted secretary protein with a small molecular weight, AY358935 might have important functions in cellular proliferation and anti-viral innate immune regulation.

Amino Acid Sequence ; Chromosomes, Human, Pair 11 ; genetics ; Computational Biology ; methods ; Humans ; Molecular Sequence Data ; Proteins ; genetics ; metabolism ; Sequence Homology, Amino Acid ; Software ; Vesicular Stomatitis ; metabolism