Objective To understand the underlying mechanism of mites-induced pediatric asthma by bioinformatic analysis on speciifc microRNA (miRNA) array and target gene screening. Methods This is a case control study of 62 pairs of dust mites-induced asthma children with age and gender matched healthy controls. Twelve pairs were randomly selected for miRNA array. The abnormal expression of miRNAs was compared between asthma and control children. The results were validated by RT-qPCR and bioinformatic analysis in remaining pairs of children. Results Six miRNAs (miRNA-151a-5p, 625-5p, 126-3p, 513a-5p, 27b-3p, 22-3p) were signiifcantly down-regulated more than two folds in dust mites-induced asthma children than those in controls. The enriched bioinformatics analysis showed that these miRNAs and their target genes CBL, PPARGC1B, ESR1, ONECUT2, EGFR, SYK, and STAT1 were related to inlfammatory cytokine signaling pathway. Conclusion It is suggested that miR-22-3p, 513a-5p, 625-5p, 27b-3p, and miRNA-target genes form a network through co-regulation to target genes to participate dust mites-induced asthma in children.

Objective To explore the success rate and the risk of establishment of the acute myocardial infarction model between the beagle dogs and the mini-pigs by interventional technique ,further to provid theoretical basis for choose a more suitable animal model .Methods 6 dogs and 6 mini-pigs were anaesthetized ,then underwent the coronary arteriography via femoral artery .After is-chemic preconditioning the coronary balloon was inflated to occlude the middle left anterior descending coronary for 180 minutes . The electrocardiogram was examined throughout the operation and the pathological sections were examined until the animals were executed one week later .Results All beagle dogs survived ,while 1 case of mini-pigs dead(1/6) .There was 1 cases(1/6) of beagle dogs had acute myocardial infarction ,while 5(5/6)cases in mini-pigs .All mini-pigs had malignant arrhythmia(6/6) but never seen in beagle dogs .The time needed for building a model was similar between the two groups ,the difference had no statistical signifi-cance(P>0 .05) .Conclusion The risk of establish myocardial infarction model in mini-pigs is higher than beagle dogs ,but the suc-cess rate is still high ,it might be the better choice .

Objective:To express the glycoprotein D of herpes simplex virus type 2 (gD2) in the insect cells,and to determine its immunogenicity.Methods:HSV-2 genome was used as the template for amplification of gD2 extracellular domain fragment gene by PCR.The PCR product was inserted into the vector Bacmind,and the constructed recombinant plasmid gD2-Bacmind was transfected into the sf9 cells to package the recombinant baculovirus.The Sf9 cells were infected by recombinant baculovirus seed derived from the forth passage(P4),the titer of P4 recombinant baculovirus was detected by a plaque assay and the expression of recombinant protein gD2 was determined by Western blotting method.The supernatant of infected cells was collected and purified by Ni-NTA affinity chromatography to obtain the target protein gD2,the purified gD2 protein was used to immunize the BALB/c mice in 0, 2, 4 weeks (gD2 group),and PBS was used as negative control(PBS group);the titers of gD2 specific IgG in serum were detected by ELISA assay.Results: The PCR analysis and sequencing results proved that gD2-Bacmind was constructed correctly.The titer of recombinant baculovirus was 2.0×109 pfu·mL-1,the purified gD2 was about 37 000 with expectation,the percentage of gD2 in total protein was 90%.The average value of Log10 of titer of gD2 specific IgG in serum detected by ELISA assay in gD2 group at the sixth week was 4.34,and there was significant difference compared with PBS group(P<0.01).Conclusion: The gD2 expressed by insect-baculovirus expression vector system has the immunogenicity and can be selected as candidate protein for HSV-2 vaccine.

Objective To investigate the expression of DNA methyltransferase 2 (DNMT2) and 3a (DNMT3a) in the epidermis of patients with psoriasis vulgaris.Methods Between March 2009 and December 2010,46 patients with psoriasis vulgaris were enrolled from the Department of Dermatology,Hospital for Skin Diseases,Chinese Academy of Medical Sciences and Peking Union Medical College,and the Department of Dermatology of Yixing People's Hospital,and 28 healthy controls were enrolled from the Department of Dermatologic Surgery,Hospital for Skin Diseases,Chinese Academy of Medical Sciences and Peking Union Medical College.Real-time quantitative PCR was performed to determine the mRNA expression of DNMT2 and DNMT3a in the epidermis of the lesional and nonlesional skin of patients with psoriasis vulgaris,as well as in the epidermis of normal skin of the healthy controls.Results The mRNA expression of DNMT2 (expressed as 2-△△Ct) in the lesional skin,non-lesional skin of the patients and normal skin of the healthy controls was 0.62 ± 0.02,0.36-± 0.05 and 0.15 ± 0.11,respectively.The mRNA expression of DNMT2 was significantly higher in the lesional skin than in the non-lesional skin of the patients (t =6.23,P ＜ 0.01),and higher in the non-lesional skin of the patients than in the normal skin of the healthy controls (t =7.33,P ＜ 0.01).Additionally,the mRNA expression of DNMT3a was significantly higher in the lesional skin (0.85 ± 0.03) than in the non-lesional skin (0.43 ± 0.04) of the patients (t =5.66,P ＜ 0.01),and higher in the non-lesiona] skin of the patients than in the normal skin of healthy controls (0.18 ± 0.09,t =8.62,P ＜ 0.01).Conclusion Both DNMT2 and DNMT3a mRNA were abnormally expressed in the epidermis of patients with psoriasis vulgaris.

Objective: To investigate the inhibitory effect of T-2 toxin on proliferation of the human hepatocellular carcinoma HepG2 cells and its promotion effect of apoptosis.Methods:The HepG2 cells in the logarithmic phase were selected and divided into control group (without T-2 toxin)and experimental groups (given 0.25,2.50,25.00,250.00 and 2500.00 μg·L-1 T-2 toxin).After 24 h treatment,the morphology of cells was observed under inverted microscope;the inhibitory rate of proliferation of cells was determined by MTT assay;the cell cycle and apoptotic rate of cells were analyzed by flow cytometry;Hochest 33258 staining was used to observe the apoptotic morphology of cells;the activity of caspase-3 in HepG2 cells was detected.Results:After treated for 24 h,the inverted microscope observation results showed that the number of the cells in experimental groups was decreased significantly and the cells shrank and deformed.The MTT results showed that compared with control group,the inhibitory rates of proliferation of the cells in experimental groups were increased (P <0.01).The flow cytometry results showed that compared with control group, the percentage of the cells in SubG1 phase in experimental group was significantly increased,and the apoptotic rates of the cells in experimental groups were significantly increased.The Hoechest 33258 staining results showed that the chromatin condensation was observed in the cells in experimental groups,and the nuclei were dense and stained.Compared with control group,the activities of intracellular caspase-3 of the cells in experimental groups were significantly increased (P < 0.01 ). Conclusion:T-2 can inhibit the proliferation of human hepatocellular carcinoma HepG2 cells,and induce the apoptosis.