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 construct DNA vaccine expressing Mycobacterium tuberculosis(Mtb) immunodominant antigen Ag85A and analyze its anti-tuberculosis T cell responses in BCG primed-mice after DNA vaccination boosting.Methods The coding gene of Ag85A mature fragment was amplified by PCR with H37Rv genomic DNA as template,and then cloned into the eukaryotic expression vector pVAX1 to construct Ag85A DNA vaccine.After purification,Ag85A DNA vaccine was injected intramuscularly twice in BCG primed-mice with BCG vaccination and DNA vaccination alone as control.Eight weeks post-vaccination,spleen lymphocytes were separated and were then used to analyze Mtb antigen specific effector T cell response and polyfuntional IFN-γ/TNF-α/IL-2 secreting CD4+ T cell frequencies and intensities,and CD8+T cell responses by IFN-γ ELISPOT assay and intracellular staining,respectively.Results Compared to BCG vaccinated-and DNA vaccinated-mice,Ag85A DNA boosting not only enhanced significantly BCG primed-mice IFN-γ+TNF-α+IL-2+,IFN-γ+ IL-2+,TNF-α+IL-2+ and IL-2+ CD4+ T cell frequencies and IL-2 secretion,but also improved significantly IFN-γ-secreting and IL-2-secreting CD8+ T cell frequencies.Condusion Ag85A DNA vaccine was constructed successfully and was demonstrated to enhance significantly BCG primed-mice Mtb antigen specific CD4+ and CD8+ T cell responses when boosting,which is beneficial to improve BCG immunogenicity and its waning immune protection against Mtb.

Large amount of data including chemical composition and size information of individual particles would be generated in the measurement of aerosol particles using atmospheric aerosol time-of-flight mass spectrometry ( ATOFMS ) . Our home-made ATOFMS was used to measure the indoor individual aerosol particles in real-time for 24 h, and the obtained mass spectrometric data were clustering analysis by self-organizing map ( SOM ) because of its ability of vector quantization and data dimensionality reduction. 20 classification results were got which includedCalcium-Containing,Salt+Secondary particles,Secondary particles,Organic Amines,K+-Rich Organics andSoil particles, etc. Compared with previous mass spectrometric methods, SOM is a natural visualization tool, more classification results can be obtained. This classification information would be useful to assess the response and toxicity of atmospheric aerosol particles and identify the origin of atmospheric aerosol particles.

Objective To investigate the effect of portal hypertension (PH) on the activity of local renin angiotensin system(LRAS). Method The expression of local renin and local angiotensinogen mRNA in liver,splenic artery and vein of PH patients was detected with RT-PCR. Result Expression of local renin mRNA in the liver in control group was 0.19?0.12, significantly lower than that of splenic artery (0.45?0.12) and splenic vein (0.39?0.12) respectively(P

Parathyroid carcinoma is a rare sporadic disease.Clinical manifestations include hypercalcemia,increased urinary calcium and urinary calculus,osteoporosis,and pathologic fracture.While this patient complained of pollakiuria,urgency,and painful urination,thus might lead to misdiagnosis or missing of diagnosis of parathyroid carcinoma.This article is herewith presented to call attention to discovery,diagnosis,and treatment of parathyroid carcinoma.

Objective Toll-like receptor ( TLR) gene family is the most important pathogen recognition receptor and animal experiment have found TLR4 mice is inclined to infect aspergillosis ( IA) .The study was to investigate the variation of TLR4 gene in Chinese Han nationality and its relation with the susceptibility of IA. Methods 25 patients diagnosed with proven IA from June 2011 to December 2012 in our hospital were enrolled, among which 17 were males.Their average age was 52.4 ±12.3, and 12 pa-tients had underlying diseases, the others had no underlying diseases.The control group consisted of 103 normal persons, among which 70 were males.Their average age was 56.0 ±17.2.All of the subjects were Chinese Han population.DNA was extracted from periph-eral blood.Polymerase chain reaction ( PCR) was applied to amplify the coding sequence of TLR4 gene followed by sequencing.The sequencing result was compared with TLR4 coding sequence in NCBI GenBank along with the analysis on amino acid change caused by genetic mutation and its effect on protein function.Comparison analysis was made on genetic mutation rate between IA group and con-trol group. Results Two missense mutations,TLR4 219 C>G and 1420 C>T, were identified in TLR4.The prediction result of protein structure showed 219 C>G resulted in the change of functional area for TLR4 to recognize pathogen and 1420 C>T caused no change in domains.The variation rate of TLR4 in IA group was 8%( 2/25 ) and both the patients had no underlying diseases. No mutation has been founded in control group and the difference between two groups was of statistical significance (P=0.037). Conclusion Two missense mutations (219 C>G and 1420 C>T) have been detec-ted in encoding area of TLR4 gene of IA patients.Variation in the cod-ing region of TLR4 gene may increase the susceptibility to IA.

Objective To observe the protection of human lens epithelial cells(HLEC) by different polar alkaloids extracted from Herba Dendrobii(HD).Methods We extacted the Herba Dendrobii powder with ethanol,and then treated the extract with falling-film concentration,acidification,salting out,chloroform extraction,and washing with water.Different polar alkaloids were extracted from HD after the above treatment.The protective effect of HD alkaloids was observed on HLEC,which were cultured with DMEM medium containing 10% fetal calf serum.Ten groups were set up for the experiment: normal control group,model group,high-and low-dose water-soluble alkaloids groups,high-and low-dose fat-soluble alkaloids groups,high-and low-dose low-polar alkaloids group,and high-and low-dose weak-polar alkaloids groups.The high-dose dosage of the alkaloids was 25.0 ?g/L and low-dose dosage was 12.5 ?g/L.Methyl thiazolyl tetrazolium(MTT) assay was used to evaluate the proliferation of HLEC under the different conditions of interventions.Results The single-factor experiments showed that the highest extracting rate of HD alkaloids was obtained under the conditions of extracting the powder with 80% ethanol for 3 times and for 3 hours.The results of protective experiment showed that the proliferation of HLEC in the model group was inhibited by hydrogen peroxide(H2O2),and the inhibitive rate was lower in low-dose fat-soluble alkaloids group than that in the model group(P

Prediction of RNA secondary structures including pseudoknots is a difficult topic in RNA field. Current predicting methods usually have relatively low accuracy and high complexity. Considering that the stacking of adjacent base pairs is a common feature of RNA secondary structure, here we present a method for predicting pseudoknots based on covariance with stacking and minimum free energy. A new score scheme, which combined stacked covariance with free energy, was used to assess the evaluation of base pair in our method. Based on this score scheme, we utilized an iterative procedure to compute the optimized RNA secondary structure with minimum score approximately. In each interaction, helix of high covariance and low free energy was selected until the sequences didn't form helix, so two crossing helixes which were selected from different iterations could form a pseudoknot. We test our method on data sets of ClustalW alignments and structural alignments downloaded from RNA databases. Experimental results show that our method can correctly predict the major portion of pseudoknots. Our method has both higher average sensitivity and specificity than the reference algorithms, and performs much better for structural alignments than for ClustalW alignments. Finally, we discuss the influence on the performance by the factor of covariance weight, and conclude that the best performance is achieved when lambda1 : lambda2 = 5 : 1.
Algorithms ; Base Pairing ; Base Sequence ; Computational Biology ; methods ; Molecular Sequence Data ; Nucleic Acid Conformation ; RNA ; chemistry ; genetics ; Sequence Analysis, RNA

OBJECTIVETo study the anti-cataract effect of gigantol combined with syringic acid and their action mechanism.

METHODH202-induced lens oxidative injury in vitro rat model was establish to observe the impact of gigantol combined with syringic acid on lens transparency under a dissecting microscope. D-galactose-induced cataract rat model was established to observe the impact of gigantol combined with syringic acid on lens transparency under a slit-lamp. UV spectrophotometry was adopted to detect the inhibitory activity of gigantol combined with syringic acid against AR. Molecular docking method was used to detect binding sites, binding types and pharmacophores of gigantol combined with syringic acid in prohibiting aldose reductase.

RESULTBoth in vitro and in vivo experiments showed a good anti-sugar cataract activity in the combination of gigantol and syringic acid and a better collaborative effect than single component-gigantol and syringic acid and positive control drug Catalin. Molecular docking and dynamic simulation showed their collaborative AR-inhibiting amino acid residue was Asn160 and the major acting force was Van der Waals' force, which formed common pharmacophores.

CONCLUSIONGigantol combined with syringic acid shows good anti-cataract, their action mechanism is reflected in their good collaborative inhibitory effect on AR.

Aldehyde Reductase ; antagonists & inhibitors ; Animals ; Bibenzyls ; Cataract ; drug therapy ; enzymology ; Drug Synergism ; Female ; Gallic Acid ; analogs & derivatives ; pharmacology ; Guaiacol ; analogs & derivatives ; pharmacology ; Humans ; In Vitro Techniques ; Lens, Crystalline ; drug effects ; enzymology ; Male ; Rats ; Rats, Wistar