In recent years , a series of agriculture biosecurity accidents have occurred ,such as mad cow disease , foot-and-mouth disease and avian influenza , which has aroused worldwide , concern over agriculture bioterrorist attacks .This paper comprehensively analyzes the history and impact of agriculture bioterrorism , the current status of international agricul-ture bioterrorism research , and important issues to be taken into account in future efforts to deal with agriculture bioterrorism .

Objective To develop a BP neural network to differentiate between ventricular fibrillation( VF) and non-VF rhythms.Methods Eighteen metrics were extracted from the ECG signals.Each of these metrics respectively characterized each aspect of the signals, such as morphology, gaussianity, spectra, variability, and complexity.These metrics were regarded as the input vector of the BP neural network.After training, a classifier used for VF and non-VF rhythm classification was obtained.Results and Conclusion The constructed BP neural network was tested with the databases of VFDB and CUDB, and the accuracy was 98.61%and 95.37%, respectively.

Objective We increase the soluble expression of artificial tandem hybrid ubiquitin binding domains ( ThUBD) in prokaryotic cytoplasm of Escherichia coli BL21 ( DE3 ) , which offer an effective and special profiling for ubiquitin conjugates( UbC) .Methods Codon optimization of the ThUBD was performed, followed by analysis of codon relative adaptiveness based on relative frequency of synonymous codon ( RFSC) of E.coli.Further induced expression and yeast ubiquitin conjugate enrichment quantified the soluble ThUBD-S and tested the ability to bind UbC.Results The statistical result showed that the percentage of codon of the highest usage frequency was increased from 48%to 75%, and codon adaptation index( CAI) was increased from 0.63 to 0.88 after codon optimization, which might suggest a higher expression of the ThUBD in E.coli BL21 (DE3).The subsequent SDS-PAGE indicated that the soluble target protein was increased four times, which accounted for 13.06%of total cell lysis.Further ubiquitinated proteome of yeast demonstrated that the ability to bind and enrich UbC of optimized ThUBD-S did not change compared with original ThUBD.Conclusion The expression of ThUBD-S can quadruple after codon optimization.At the same time, codon optimization does not impact its soluble expression and the ability to bind UbC.

Objective To explore the construction method of semantic types of military medical ontology and to establish a framework for the semantic types of the top-level ontology of military medicine in order to lay the foundation for the construction of military medical ontology and provide support for knowledge organization, knowledge disclosure and knowledge service of military medical information resources.Methods Using the method of document measurement, the core concept set of military medicine was constructed.On the basis of the framework of the top-level semantic types of Unified Medical Language System(UMLS), the semantic type framework was completed by the combination of bottom-up and top-down modes, using semantic analysis and expert consultation.Results By classifying the same kind of knowledge points of military medicine into one semantic type, the hierarchical structure of the semantic type framework of the top-level ontology of military medicine was obtained.Conclusion The proposed method is effective and practical for the construction of the semantic types of military medical ontology.The established semantic type framework of the top-level ontology of military medicine could reconstruct military medical knowledge and provide the basic framework for the establishment of the semantic relationships of the top-level ontology of military medicine.

Objective To determine the Golgi dispersal in radiation damaged cells and the protective effect of vanillin derivatives.Methods Immunofluorescence, cell cycle analysis of flow-cytometry,Western blot,and clone formation were used.Results Immunofluorescence observation showed that the Golgi dispersal caused by 2 Gy 60 Coγ-ray was significantly increased in a dose-dependent manner in the range of 4-10 Gy as was demonstrated by the fact that the Golgi area was significantly increased. When the irradiated cells were treated with the radioprotective agent VND3207, a vanillin derivative,the Golgi dispersal induced by radiation was significantly reduced.The radiation-induced Golgi dispersal was also displayed in a pattern of time-course after irradiation in the HeLa cells, and persisted at least to 36 h post-irradiation. Cell cycle test results indicated that the Golgi dispersal was not associated with the G2/M arrest triggered by radiation-induced DNA damage response.VND3207 could promote cell survival by plate colony formation assay.Conclusion The Golgi dispersal can be caused byγ-ray irradiation in a dose-and time-dependent manner, and VND3207 can provide a good protection against radiation injury associated with inhibited Golgi dispersal.

Objective To construct the eukaryotic expression vector of PRDX3 labeled with FLAG tag and to study its localization in human tongue cancer cell line SCC15.Methods PRDX3 gene was obtained from the breast library by PCR and cloned into PCDH vector to construct PCDH-FLAG-PRDX3.The plasmid was transiently transfected into 293T cells and the expression was detected by Western blot.Subcellular localization was detected by cellular immunofluorescence.Results The result of double digestion and sequencing showed that PCDH-FLAG-PRDX3 eukaryotic expression vector was constructed.The expression of FLAG-PRDX3 in human 293T cells was positively confirmed by Western blotting.In human tongue cancer cell line SCC15, the result of cellular immunofluorescence showed FLAG-PRDX3 was located in the cytoplasm rather than in the nucleus.Conclusion PRDX3 eukaryotic expression vector labeled with FLAG tag is constructed successfully, which is located in cytoplasm in human SCC15 cells.Construction and identification of PRDX3 could shed light on the function and mechanism of PRDX3 in tongue cancer.

Objective To observe the inhibitive effect of electro-acupuncture (EA)at Zusanli points (ST36)on inflammatory mediators of postoperative intra-abdominal adhesions and study the relationship between EA and cholinergic anti-inflammatory pathway.Methods Forty-eight male Wistar rats were divided into 6 groups (each =8):Group A (control),Group B(abdominal adhesions model),Group C (abdominal adhesions plus EA),Group D(sham acu-point control),Group E (abdominal adhesions plus α-bungarotoxin )and Group F (abdominal adhesions plus EA after α-bungarotoxin).Animal models of abdominal adhesion were produced by Chiang’s path.Bilateral Zusanli points (ST36) and shame acupoints were electro-acupunctured at a constant voltage for 1 hour while rats were awake.The ɑ-BGT(1 μg/kg)was injected into the abdominal cavity after surgery.All the rats were sacrificed on the 3rd day,and the levels of inflammatory mediators (TNF-ɑ,NO and NOS)in tissues were evaluated.Results Three days after surgery,the damaged cecum of abdominal adhesion groups developed obvious edema that did not adhere with other tissues.Compared with sham control,the abdominal adhesion resulted in significant elevation of inflammatory mediators (TNF-ɑ,NO and NOS).EA at Zusanli points obviously lowered the elevated levels of inflammatory mediators (P <0.01 and P <0.05).EA at Zusanli points following the injection of ɑ-BGT showed less anti-inflammatory effect(P <0.01).Conclusion EA at Zusanli points significantly lowers the elevated levels of inflammatory mediators after abdominal adhesion challenge.The activation of cholinergic anti-inflammatory pathway might be one of the mechanisms by which Zusanli points exert anti-inflammatory effects.

Objective To investigate the clinical value of axillary ultrasound (AUS)in the identification of axillary nodal metastasis (ALNM).Methods Two hundred and eighty-two consecutive patients with stage Tis-T2 breast cancer were prospectively enrolled between December 2013 and September 2015.All the patients underwent AUS performed by two specified senior ultrasound doctors.Sonographic features of their axillary lymph nodes (longitudinal and transverse diameters,cortical and hilar thickness,blood flow form)were collected.These patients were divided into metastatic, suspicious and non-metastatic groups based on the ultrasound features by ultrasound doctors.The diagnostic accuracy of AUS was compared with results of pathology.Univariate and multivariate Logistic regression analyses were used to evaluate the relationship between sonographic features and ALNM.The area under the ROC curve was used to assess the accuracy of the multivariate Logistic regression model.Results The sensitivity,specificity,positive and negative predictive value and accuracy of AUS were respectively 85.6%,87.1%,86.4%,86.3%,and 86.3% in the metastatic and non-metastatic groups.The Kappa value was 0.727(P <0.001).The ALNM burden in the non-metastatic group was significantly lower than in the metastatic group (1.2 vs 6.9,P <0.001).The false-negative results were found only in 16 cases,fourteen of whom had only 1,and two had 2 and 3 ALNM,respectively.Univariate Logistic regression analysis showed that maximum cortical thickness was the most significant predictive factor of ALNM(the area under the ROC curve was 0.872).Multivariate Logistic regression analysis suggested that cortical thickness and the ratio of hilar thickness to cortical thickness were predictive factors of ALNM(P <0.05).The area under the ROC curve of the multivariate Logistic regression model was 0.879 and its sensitivity and specificity were 77.0% and 85.1%,respectively.Conclusion AUS is a valuable tool for detecting ALNM.Patients with false-negative results of AUS have a lower axillary metastatic burden.Maximum cortical thickness is the most significant predictive factor of ALNM.AUS may be a potential alternative method for sentinel lymph node biopsy as axillary lymph node staging in early-stage breast cancer patients.

Objective To find out more about the population structure and clonal complex of clinical Vibrio parahaemolyticus strains in Asia.Methods Clinical Vibrio parahaemolyticus strains data were screened in Asia with complete ST and pST types from PubMLST public database,their subgroup and complex were analyzed,and the minimum spanning tree based on ST and pST types respectively was completed.Results From the database,341 items of ST and pST types of Asian clinical Vibrio parahaemolyticus strains were screened,including 157 ST,most of which were of ST3 type.Totally 214 items of data came from China (including Hong Kong and Taiwan),and covered 133 ST,most of which were of ST3 type.eBURST software was used and 17 groups and 94 singletons were found.Software STRUCTURE analysis showed that the appropriate subset number of clinical V.parahaemolyticus strains in Asia was 7,and that the average distance between samples in each subgroup was 0.9113.Conclusion Clinical V.parahaemolyticus strains in Asia show high diversity and can be subdivided into seven subgroups.ST3 type is dominating when multilocus sequence typing(MLST)is used and pST2 type is the majority by AA-MLST typing.

Objective To construct the pseudovirus containing nucleic acid(NA)fragments of Marburg virus,Zaire Ebola virus,Sudan Ebola virus,Lassa fever virus and Yellow fever virus by using a lentiviral vector system in order to provide a reference standard for the detection of the five viruses.Methods The gene fragments of the above five viruses were synthesized in vitro,connected into a single gene by fusion PCR technique,and cloned into lentiviral vectors with its auxiliary vector.After co-transfecting into 293T cells,the supernatants were collected on 48 h and 72 h post transfection. The naked NA was cleaned from the supernatants using DNase and RNase digestion before pseudotype virus was purified and concentrated.After the NA of the pseudotype virus,were extracted normal PCR and real-time PCR were conducted. Results Sequence analysis showed that the five target genes in vitro synthesis were properly connected and inserted into lentivirus vectors.Using the NA of the pseudotype virus as the template,both normal PCR and real-time PCR could sensitively amplify the target gene with the primers and probes of the above five,viruses respectively.The result indicated that the pseudovirus particles containing the five kinds of hemorrhagic fever virus target genes were successfully packaged. Conclusion The pseudovirus particles containing gene fragments of five viruses are constructed,which can be used as a common reference standard for NA detection.