ObjectiveTo compare three types of acute Mycobacterium tuberculosis infection mouse models established through different infection routes and to set up the theoretical basis for further developing,selecting and applying these animal model in the tuberculosis-related research.MethodsStandard strain of Tubercle bacillus H37Rv was diluted to 1 × 106 colony forming unit (cfu)/mL.The mice were infected with the bacteria through different routes including intravenous injection,intranasal administration and inhalation of bacteria aerosol.Six weeks after the infection,the mice were euthaniz ed and necropsied. The lung tissues were collected and gross changes were observed.The colony counting was performed and the lung tissues were assessed by HE staining,acid fast staining.The e xpression level of tumor necrosis factor (TNF)-α per unit area in lung tissue was detected by immunohistochemistry. The data were analyzed by t test. Results The amounts of Mycobacterium tuberculosis in lung tissues of mice in inhalation group,intranasal administration group and intravenous injection group were (6.290±0.028),(6.150±0.021) and (6.120±0.008) lg cfu/mL,respectively; while no Mycobacterium tuberculosis was detected in control group. The difference between infection group and control group was statistically significant (t =3.762,P＜0.01),while there were no significant differences among infection groups with different infection routes (P＞0.05).According to the results of gross observations and histological assessment,the pathological changes were observed and red tubercle bacillus was detected by acid-fast staining in the lung tissues of all the mice in infection group.The results of immunohistochemistry showed that the expression levels of TNF-α per unit area were as follows:intravenous injection group (0.049 × 106 )＜intranasal administration group(0.759×106) ＜ inhalationgroup(1.042×106), whichwere statistically different (t =2.504,P＜ 0.05).ConclusionInhalation of bacteria aerosol may be the most efficient method to establish tuberculosis infection mouse model compared to intravenous injection and intranasal administration.

Objective To compare and analyze the differences and characteristics of three strain mouse models in-fected by influenza virus aerosol inhalation, and provide the reference for choosing the appropriate infection model in the re-search of pathogenesis of influenza and the development of vaccines and drugs.Method C57BL/6, BALB/c and ICR mice were infected with A/Puerto Rico/8/34 (H1N1) virus strain by aerosol inhalation.The symptoms and body weight of mice were observed every day.At 3, 7, 14 days after infection, the mice were sacrificed.The lungs of mice were weighed, then virus assay and pathological observation were carried out.Results The three strains of mice were infected.The sur-vival rate in the C57BL/6 mice was lower than those in the BALB/c and ICR mice.The lung index and viral load of C57BL/6 mice were significantly higher than those of ICR mice ( P<0.05) at 3 days after infection.The pathological changes of C57BL/6 mice were also more obvious than other two strains.Compared with other two mouse strains, the weight recovery of BALB/c mice was the slowest.The survival rate in BALB/c mice was higher than that of C57BL/6 mice and lower than that of ICR mice.The lung index and viral load were not significantly different among the three strains of in-fected mice.The pathological changes among the three strains of infected mice were similar, but the degrees of pathological changes in the BALB/c mice were milder than in the C57BL/6 mice and worse than in the ICR mice.Compared with other two mouse strains, the process of disease is similar, but the body weight, mortality, lung index, viral load, and the micro-scopic pathological changes were lighter in the ICR mice than in the other two strain mice.Conclusions The three strain mouse models can be established by influenza virus aerosol inhalation, but showing different characteristics.Appropriate strain mice can be chosen to build model according to different research purpose in the experiment.

Objective To establish an in vivo imaging method of normal or tumorous liver in mice by using a new type nanoparticle contrast agent, ExiTron nano 12000, coupled with micro-CT imaging.Methods Six 6-8-week old male C57BL/6J mice were randomly divided into group A and group group B, by intravenous injection of 50μL and 100μL Ex-iTron nano 12000, respectively.In vivo Micro-CT scans were performed before contrast agent injection, 3 minutes, 24 hours, 7, 14, 28 and 56 days after injection.To determine which dose is suitable for long-term studies, gray scale value a-nalysis was performed on selected region of interest ( ROI) in the left lobe and right anterior lobe of the liver, and the chan-ges of liver tissue contrast was monitored after ExiTron nano 12000 injection.Three male HBV transgenic mice bearing liver tumors ( group C) were intravenously injected with the determined dose of ExiTron nano 12000 and were monitored by mi-cro-CT scans as above described.At 56 days after ExiTron nano 12000 injection, the mice were sacrificed and liver sam-ples were taken for histological analysis.Results Cross-sectional images taken at various time points and the average gray scale value ( AGSV) analysis in the mouse liver revealed that the AGSV peaked at 24 hours after injection of contrast rea-gent and good contrast still presented in the livers within 56 days of observation for both groups, though group B showed a significantly higher contrast than group A (P＜0.01).Those data indicated that the dose of group B (100μL) was better to maintain ExiTron nano 12000 in the liver of mice for a long time.Contrast-enhanced by 100μL of ExiTron nano 12000, the liver tumor nodules in the mice of group C could be clearly delineated by Micro CT imaging during a 56 days observa-tion.Histological analysis revealed atypical hyperplasia, enlarged nuclei with hyperchromasia and cell necrosis in the tumors.Conclusions An in vivo imaging method was established to non-invasively visualize mouse liver using micro-CT combined with nanoparticle-based contrast agent and this technology may be applied to a live imaging of murine primary liv-er tumors.

Object To establish an induced mouse model of type 2 diabetes mellitus, and compare it with db/db mouse model of spontaneous type 2 diabetes.To evaluate the two mouse models objectively, and provide an experimental basis for the choice of animal model and its practical application in diabetes studies.Methods A mouse model of induced type 2 diabetes was established by feeding C57BL/6J mice with high-fat and high-sugar diet for four weeks and taking daily intraperitoneal injections of streptozotocin ( STZ) for consecutive 3 days.Four weeks after infection, the gross appearance of kidney and liver of the mice was assessed, glucose tolerance was tested, serum biochemical indices and expression of se-rum IL-2, IL-4, IL-6, IFN-γ, TNF-α, IL-17, and IL-10 were assayed, and were compared with those of the db/db mouse models of spontaneous type 2 diabetes.Results Obvious differences were found in the kidneys and liver gross appearance of the two types of mouse models and the control group.Both the two groups showed significant differences in the blood glu-cose levels at each time point (P＜0.05) and low glucose tolerance function, but there were no significant differences in blood glucose levels of the two types of mouse models.Compared with the control group, the serum biochemical indices GLU, GHOL and LDLC of the two types of mouse models were significantly increased (P＜0.05).Meanwhile, the blood lipid level of the mouse model of induced type 2 diabetes was higher than that of the db/db mouse models of spontaneous type 2 diabetes.In comparison of immune indices, except IL-2,the serum cytokine levels of the two types of mouse models were significantly higher than those of the control group (P＜0.05).Moreover, the serum cytokine levels of db/db mice were higher than those in the mouse models of induced type 2 diabetes, and the IL-6, IFN-γ, TNF-αalso had obvious differences.Conclusions Both the two types of mouse models of type 2 diabetes successfully simulate the human diabetes to some extent, but there are still certain differences according to different etiology of diabetes.We would suggest that peo-ple may take our data as reference and chose appropriate mouse models according to the requirement of their research.

In recent years, the MYB-related gene family has been found pivotal in plant growth and development. MYB-related gene family in Angelica dahurica var. formosana was systematically investigated based on "Chuanzhi No. 2" through transcriptome database search and bioinformatics and the temporal and spatial expression patterns were analyzed through real-time fluorescence-based quantitative polymerase chain reaction(PCR). The results showed that 122 MYB-related proteins family were identified, mainly including the unstable hydrophilic proteins with good thermal stability. Most of the proteins were located in nuclei. The majority of the proteins had the structures of random coil and α-helix. Five MYB-related proteins family of A. dahurica var. formosana had membrane-binding domains. The conserved domain analysis of MYB-related proteins family of A. dahurica var. formosana showed that the MYB domains of genes in five subgroups, similar to 2 R-, 3 R-, and 4 R-MYB proteins, contained three evenly distributed Trp(W) residues in the MYB repeat sequence. The phylogenetic analysis of MYB-related proteins family in A. dahurica var. formosana and Arabidopsis thaliana showed that the MYB-related members were unevenly distributed in five subgroups, and A. thaliana and A. dahurica var. formosana had almost the same number of genes in the CCA1-like subgroup. There were differences in the number, type, and distribution of motifs contained in 122 encoded proteins. Transcription factors with similar branches had similar domains and motifs. The expression pattern analysis showed that the transcription factors AdMYB53, AdMYB83, and AdMYB89 responded to hormones to varying degrees, and they were highly expressed in leaves and responded quickly in roots. This study lays a foundation for further investigating the function of MYB-related transcription factors of A. dahurica var. formosana and solving the corresponding biological problems such as bolting early.
Angelica/chemistry* ; Animals ; Computational Biology ; Gastropoda ; Phylogeny ; Plant Leaves ; Plant Proteins/genetics* ; Transcription Factors/genetics*

LysR-type transcriptional regulators are involved in the regulation of numerous cellular metabolic processes in Klebsiella pneumoniae, leading to severe infection. Earlier, we found a novel LysR family gene, named kp05372, in a strain of K. pneumoniae (designated GPKP) isolated from forest musk deer. To study the function of this gene in relation to the biological characteristics of GPKP, we used the suicide plasmid and conjugative transfer methods to construct deletion mutant strain GPKP-Δkp05372; moreover, we also constructed the GPKP-Δkp05372+ complemented strain. The role of this gene was determined by comparing the following characteristics of three strains: growth curves, biofilm formation, drug resistance, stress resistance, median lethal dose (LD₅₀), organ colonization ability, and the histopathology of GPKP. Real-time polymerase chain reaction (RT-PCR) was used to test the expression level of seven genes upstream of kp05372. There was no significant difference in the growth rates when comparing the three bacterial strains, and no significant difference was recorded at different osmotic pressures, temperatures, salt contents, or hydrogen peroxide concentrations. The GPKP-Δkp05372 mutant formed a weak biofilm, and the other two strains formed medium biofilm. The drug resistance of the GPKP-Δkp05372 mutant toward cephalothin, cotrimoxazole, and polymyxin B was changed. The acid tolerance of the deletion strain was stronger than that of the other two strains. The LD₅₀ values of the wild-type and complemented strains were 174-fold and 77-fold higher than that of the GPKP-Δkp05372 mutant, respectively. The colonization ability of the GPKP-Δkp05372 mutant in the heart, liver, spleen, kidney, and intestine was the weakest. The three strains caused different histopathological changes in the liver and lungs. In the GPKP-Δkp05372 mutant, the relative expression levels of kp05374 and kp05379 were increased to 1.32-fold and 1.42-fold, respectively, while the level of kp05378 was decreased by 42%. Overall, the deletion of kp05372 gene leads to changes in the following: drug resistance and acid tolerance; decreases in virulence, biofilm formation, and colonization ability of GPKP; and regulation of the upstream region of adjacent genes.
Animals ; Bacterial Proteins/physiology* ; Biofilms ; Deer/microbiology* ; Drug Resistance, Bacterial ; Female ; Klebsiella Infections/pathology* ; Klebsiella pneumoniae/growth & development* ; Male ; Mice ; Transcription Factors/physiology*

Objective To evaluate the regional cerebral metabolic changes in different episodes by magnetic resonance spectroscopy (MRS) after explosive brain injury in rabbits. Methods Fortyfive New Zealand white rabbits were randomly divided into eight groups, ie, normal control group( 10 rabbits) and trauma group (35 rabbits). The explosive injury in trauma group was induced by explosion of 600 mg TNT equivalent of paper detonators at 6.5 cm above the rabbit brain. The rabbits in trauma group was divided into 1,6, 12, 24 hours, 3, 7, 14 days subgroups (6 rabbits per group). The survival rate was observed at different time points after explosive injury. The MRS was used to detect the regional cerebral metabolic changes including N-acetylaspartate (NAA)/creatine (Cr) ratio and choline(Cho)/Cr ratio as well as evolution of blast injuries over time. Results The rabbits survived for overseven days in the trauma groups, with typical brain contusion manifested by pathological and conventional MRI. Compared with the normal control group, the NAA/Cr ratio was markedly decreased at one hour after injury, slightly rose again at 24 hours and fell again after seven days. The Cho/Cr ratio was markedly increased at one hour after injury, slightly fell again at 12 hours and rose again at three days after injury.Conclusions MRS can manifest the regional cerebral metabolic changes of rabbits with explosive injury at different time points and hence provide a theoretical basis for understanding the local tissue changes and determining the type of tissue damage after blast injury.

Objective To prepare a novel radiolabeled FR?positive tumor targeting agent 99 Tcm?( HYNIC?NHHN?FA) ( EDDA) and evaluate its biological properties. Methods FA derivative FA?NHHN?HYNIC was synthesized and radiolabeled with 99 Tcm using EDDA as a coligand. The radiochemical purity, octanal/water partition coefficient and in vitro stabilities of the complex were studied after purified by HPLC. In vitro cellular uptakes were performed on FR?positive KB cells ( human oral epidermoid carcinoma cells) . Biodistribution and microSPECT/CT imaging were investigated on normal Kunming mice and nude mice bearing KB tumors, respectively. Results The radiochemical purity of the complex was over 95% after pu?rified by HPLC. It displayed high stability both in saline and in serum. It also exhibited high specific FR binding in FR?positive KB cells in vitro. The binding ratio was (6.76±0.60)%1 h after incubation, and de?creased to (0.24±0.02)% after adding excessive FA. The results of biodistribution showed high kidney up?take in normal mice, and the uptake reached (21.79±9.79) %ID/g 0.5 h after injection. Flank KB tumors were clearly visualized with 99 Tcm?( HYNIC?NHHN?FA ) ( EDDA ) by microSPECT/CT imaging at 2 h postinjection, and the uptake could be inhibited by excessive FA. Conclusions 99 Tcm?( HYNIC?NHHN?FA) ( EDDA) exhibits good pharmacokinetic properties, suggesting its potential as a promising FA targeting agent for tumor imaging.

Objective To investigate the value of multi-slice spiral CT(MSCT) in diagnosis of laryngocarcinoma in early stage.Methods MSCT data of 28 patients with laryngocarcinoma confirmed pathologically were analyzed retrospectively.Images quality was evaluated and the results obtained with various windows and CT virtual laryngoscopy (CTVL) were compared.Results 20 cases(69%) could be displayed with conventional soft tissue windows,24(81%) could be demonstrated with lung windows and 27(93%) could be demonstrated with CTVL.Conclusion MSCT can effectively demonstrate laryngocarcinoma,and can be applied routinely in examination of laryngocarcinoma.

Objective To evaluate the forensic application of TE-MAGS technology based on magnetic beads kit on TECAN pipetting platform and establish the automated DNA extraction system of case work samples. Methods Sensitivity test: 10 different DNA samples from 0.1ng to 1ng were prepared with a commercial standard DNA 9947A diluted into 200μL TES. DNA samples were purified by the TE-MAGS technology automatically on the TECAN pipetting platform and then typed using the IdentifilerTM Kit and get the profile of STR with the software GeneMapper ID-X; the power of purification was tested with a trail that purified 1ng DNA mixed with humus acid and hemachrome. Comparative test: 304 casework samples were divided into two purified by TE-MAGS technology and silicon beads respectively to compare the power of purification and the possibility of forensic utility. Results Sensitivity test: 0.3ng and more imported DNA can obtain a good quality of DNA profile compared to the lower imported DNA with dropout of STR peaks (0.1ng and 0.2ng). The power of purification of the TE-MAGS technology was not affected by humus acid and hemachrome. The comparison result between automatic TE-MAGS technology and manual silicon beads extraction methods from 304 casework samples showed that the former's success rate(50%) was higher than the latter(40.8%). Conclusion The established DNA purification method of TE-MAGS technology automatic DNA extraction system in this study was obviously advantaging at the aspect of success rate, stability, and uniformity and suited to application in the forensic utility future.