Objective: To observe the effects of interferon-induced protein 204 (p204) over-expression on apoptosis, proliferation and migration of aortic vascular adventitial ifbroblast (VAFs) in experimental rats.
Methods: Our research included in 3 groups: Iif204-Lv group, in whichVAFs were infected by Iif204-recombined lentivirus, Con-Lv group, in which VAFs carried the empty vector without virus, Blank control group, in which VAFs were untreated. VAFs proliferation was examined by MTT method, cell apoptosis was measured by lfow cytometry and the migration was detected by scratching assay and transwell chamber method. The mRNA and protein expressions of p204, p53 and p21were evaluated by real-time q RT-PCR and Western blot analysis respectively.
Results: Compared with Con-Lv and Blank control groups, Iif204-Lv group had decreased VAFs proliferation (by OD value) at 48 hours: (0.53 ± 0.05) vs (0.66±0.03) and (0.63 ± 0.06), at 72 hours: (0.89 ± 0.06) vs (1.02 ± 0.06) and (1.01 ± 0.07); distance of cell migration (by pixel): (61.00 ± 1.83) vs (74.50 ± 6.25) and (75.50 ± 7.85); number of cell migration: (61.75 ± 10.69) vs (155.25 ± 10.21) and (153.75 ± 9.40), allP<0.05. VAFs apoptosis rates were similar among different groups. Compared with Con-Lv and Blank control groups, Ifi204-Lv group presented up-regulated mRNA expressions of p204 (3.45 ± 0.15) vs (2.09 ± 0.10) and (2.06 ± 0.09); p53 (3.41 ± 0.09) vs (2.06 ± 0.07) and (2.10 ± 0.06); p21 (3.01 ± 0.08) vs (2.05 ± 0.06) and (2.11 ± 0.08), allP<0.05.
Conclusion: p204 over-expression inhibits VAFs proliferation and migration which might be partly related to the activation of p53 and p21 expression in experimental rats.

Objective To investigate the effect of interferon alpha ( IFN-α) on apoptosis of vascular smooth muscle cells ( VSMCs) in rats and the related mechanism. Methods The cells were divided into three group:group A, group B and group C.Group A was transfected with nonspecific siRNA, group B was intervened with IFN-α and transfected with nonspecific siRNA, and group C was intervened with IFN-α and transfected with IFI204 siRNA. All the cells were cultured for 48 h. The expression of P204 mRNA was determined by semiquantitative reverse transcription polymerase chain reaction (RT-PCR).P204, RAS protein levels, and phosphorylation levels of RAF and ERK were analyzed by Western blotting. The cell apoptosis was analyzed by flow cytometry with Annexin-V FITC/PI method. Results As compared with group A, the expression of P204 mRNA and protein in group B was up-regulated (P<0.05), the cell apoptosis was increased (P<0.05), in the process of the above, the expression of RAS protein was decreased ( P<0.05) and the phosphorylation levels of RAF and ERK were dropped (P<0.05).In group C, the expression levels of P204 mRNA and protein were down-regulated (P<0.05), and cell apoptosis was decreased ( P<0.05) , the expression of RAS protein and the phosphorylation levels of RAF and ERK were increased ( P<0.05) . Conclusion P204 and RAS signal pathway participates in IFN-α regulation of apoptosis of VSMCs in rats.

The E protein of dengue virus type Ⅱ was presented on ferritin of Helicobacter pylori to construct a novel dengue nanoparticle vaccine candidate,and the immunological indexes of the vac-cine were evaluated,aiming to provide new ideas for the development of dengue vaccine.The re-combinant plasmid of E-Ferritin was optimized and synthesized,and then transfected into HEK-293F cells.The recombinant protein was expressed,identified,purified and analyzed.Mice were im-munized with E-Ferritin nanoparticle vaccine by intramuscular injection on the hind limbs on the day 0,14 and 28.ELISA,neutralization test,flow cytometry and lymphocyte proliferation test were used to detect the levels of specific antibodies,neutralizing antibodies,CD3+,CD4+and CD8+T lymphocytes in spleen cells and the proliferation of spleen lymphocytes after specific stimulation.The target protein with a size of about 69 kDa was expressed in the cells with a single band.The purified protein concentration was 0.407 g/L,and the purity was 82.32％.The results from transmission electron microscopy showed that E-Ferritin protein could be recombined into a particle structure with a particle size of about 50 nm.The results of mouse immune experiments showed that E-Ferritin protein had good immunogenicity.The average specific antibody titer of E-ferritin protein in serum was 1∶92 160 after immunization 42 d.The main subclass of antibody was IgGl.The results of flow cytometry showed that E-Ferritin as an immunogen could induce higher levels of CD4+and CD8+T lymphocyte immune response.In lymphocyte proliferation test,the level of specific stimulation in the vaccine group was significantly higher than that in the non-specific stimulation group.In conclusion,the dengue virus envelope protein ferritin nanoparticle vaccine constructed in this study has good immunogenicity,which can provide reference for the de-velopment of new dengue vaccine candidates.

OBJECTIVE: To discuss the advantages and technical limitations of various molecular genetic techniques in the diagnosis of two infants featuring all-round developmental retardation. METHODS: The two patients were initially screened by using chromosomal microarray analysis (CMA). For patient 1, his parents were also subjected to CMA analysis, and the data was analyzed by using ChAS and UPD-tool software. For patient 2, methylation-specific PCR (MS-PCR) was carried out. RESULTS: Patient 1 was diagnosed with maternal uniparental disomy (UPD) type Prader-Willi syndrome (PWS) by CMA and UPD-tool family analysis. His chromosomes 15 were of maternal UPD with homology/heterology. Patient 2 was diagnosed with deletion type PWS by combined CMA and MS-PCR. CONCLUSION Correct selection of laboratory methods based on the advantages and limitations of various molecular techniques can help with diagnosis of genomic imprinting disorders and enable better treatment and prognosis through early intervention.

Target identification of bioactive compounds is important for understanding their mechanisms of action and provides critical insights into their therapeutic utility. While it remains a challenge, unbiased chemoproteomics strategy using clickable photoaffinity probes is a useful and validated approach for target identification. One major limitation of this approach is the efficient synthesis of appropriately substituted clickable photoaffinity probes. Herein, we describe an efficient and consistent method to prepare such probes. We further employed this method to prepare a highly stereo-congested probe based on naturally occurring triterpenoid betulinic acid. With this photoaffinity probe, we identified tropomyosin as a novel target for betulinic acid that can account for the unique biological phenotype on cellular cytoskeleton induced by betulinic acid.