Objective:To study the expression of vascular endothelial growth factor(VEGF) and its receptor Flt1 in the process of mandibular fracture healing.Methods:The fracture models were made in the middle of the left mandible of 25 rabbits,then the specimens from the fracture site were taken 48 hours and 1,3,5 and 8 weeks after operation respectively. The expression of VEGF and Flt1 was detected by immunohistochemistry and in situ hybridization.Results:VEGF mRNA, VEGF and Flt1 were detected in many kinds of cells in the fracture site from 48 hours to 8 weeks after fracture, and high expression of VEGF and Flt1 were detected from 1 to 3 weeks after facture.Conclusions:VEGF and Flt1 are expressed through the process of fracture healing, VEGF may play important role in the process of blood vessel rebuilding after fracture.

Aim To study IL-8 expression of U937 cells directly induced by Pseudomonas aeruginosa(PA)and its mechanism through the mitogen-activated protein kinase pathways.Methods The expressions of IL-8 mRNA in human monocytic leukemia cell lines(U937 cells)infected by Pseudomonas aeruginosa which were in differently differentiated and its protein secretion in the supernatant of culture medium were examined by the methods of RT-PCR and ELISA respectively.And these were then to be compared with those inhibited by MAPK inhibitors.Results Pseudomonas aeruginosa was able to induce IL-8 mRNA expression and their protein secretion both in U937 cells and in PMA differentiated U937 cells and had marked time and concentration dependent relations.Pretreament of U937 cells with specific inhibitors of ERK1/2(PD98059),the kinase that activated ERK1/2,p38(SB203580)could significantly diminished the PA-induced IL-8 production(P

Lactide一glycolide copolymerization with stannous octanoate as an initiator and dedecanolas regulator was carried out.The effect of reaction condition such as reaction time,temperature, initiatorconcentration, regulator and monomer ratbonpolymerization conversion,molecular weight and its distrbu-tion were investigated.The obtained results show that the finest reaction condition are initiator 0. 075%,ternperature 170℃ and reaction time 6hr.At this reaction condition,both molecular weight and polymer-ization conversion obtained are vecy high.

Objective The study aimed to analyze the clinical features and the causes of hospital death among the 5720 acute myocardial infarction(AMI)patients from Cardiology Center,Beijing Chao-Yang Hospital during the last 8 years. Methods A total of 5720 AMI patients received treatment in the Cardiology Center from January 1st ,2002 to December 31th ,2009 were retrospectively reviewed. All patients were classified according to age into 3 groups of ≤45,46 -75,and ＞ 75 years old. The morbidity,cause of death ,whether they had the PCI therapy,mortality after PCI and the impact of gender on the cause of death were observed respectively. Results The morbidity rate of male was significantly higher than female in all three groups,and the study also found that the morbidity rate of female was significantly higher in the group of ＞ 75 years old,which however was still lower than that of male. The AMI patients were more likely to accept PCI therapy,which could significantly reduce the mortality rate. The top 3 causes of death included acute heart failure(AHF),cardiogenic shock(CGS)and AHF combined with CGS. In addition,AHF caused significantly more death in female and older(＞ 75 years old)patients. Conclusions The morbidity rate of AMI patients in Beijing Chao-Yang Hospital increased year by year. And PCI therapy could reduce the mortality rate of all groups. Revascularization treatment seems to be feasible and safe for the patients older than 75 years old.

Objective To analyze the immunogenicity of selected B-cell epitopes of Epstein-Barr virus (EBV) latent membrane protein-2 (LMP2). Methods Three potential dominant B-cell epitopes of LMP2199-209, LMP2318-322 and LMP2381-391 from EBV LMP2 had been predicted using bioinformaties methods. The gene fragments of three epitopes were cloned respectively into pET32a(+) vector and transformed into E. coli strain BL21 (DE3). After identification by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting, the expression products were purified by Ni-NTA agarose affinity chromatography. BALB/c mice in immunized groups were immunized by multi-point intracutaneous injection with the three purified epitope proteins,respectively; and mice in control groups were injected with pET32a (+) protein or phosphate buffered saline(PBS), respectively. The sera from mice at week O, week 3 and week 6 of injection were collected for determination of epitope-specific antibody IgG by enzyme linked immunosorbent assay (ELISA) using epitope proteins as coating antigens. The ability of serum antibody recognizing nature EBV antigen was determined at week 6 of immunization. Results Three epitope proteins of LMP2199-209 ,LMP2318-322 and LMP2381-391 were successfully expressed in prokaryotic system. Epitopespecific antibodies IgG could be detected respectively in the sera of all immunized mice, and the levels of antibodies increased with immunized time increasing. The antibody levels in LMP2318-322 immunized group at week 3 and week 6 were significantly higher than that of pET32a (+) protein control group (F= 493.85 and 773.99, respectively; both P＜0. 05), and the antibody levels in LMP2381-391 immunized group at week 3 and week 6 were also significantly higher than that of pET32a (+) protein control group (F= 926.33 and 309.14, respectively; both P＜0.05). Antibody level in LMP2199-209 immunized group at week 6 was significantly higher than that of pET32a ( + ) protein control group (F=87.27, P＜0.05). The antibody IgG in serum from immunized mice with three epitope proteins could all recognize nature EBV antigens, especially LMP2199-209 and LMP2381-391 immunized groups.Conclusions Three possible dominant epitopes of LMP2199-209, LMP2318-322 and LMP2381-391 from EBV LMP2 are prepared by prokaryotic expression system and exhibit obvious immunogenicity, which could be used for further research of EBV infection and related tumor vaccine.

Objective To evaluate the activities of daily living (ADL) and investigate its influential factor in primary angle-closure glaucoma patients.Methods One hundred primary angle-closure glaucoma patients (acute phase) diagnosed definitely from April to October 2014 were collected.The status of sex,gender,education,income were recorded.Barthel Index (BI),Hospital Anxiety and Depression Scale (HADS) and Glaucoma Quality of Life-15 (GQL-15) were respectively used to assess the ADL,anxiety and depression symptoms and visual quality of life in primary angle-closure glaucoma patients.At the same time the factors affecting ADL of primary angle-closure glaucoma were analyzed.Results There were no dysfunction in 12 patients,46 patients with mild dysfunction,32 patients with moderate dysfunction and 10％ (10/100) patients with severe dysfunction.BI was negatively correlated with anxiety and depression score,GQL-15 results and disease course (r=-0.819-0.395,P ＜ 0.01),and positively correlated with average monthly income (r=0.453,P ＜ 0.01).There were no correlation with gender,sex and education (r=-0.159-0.172,P ＞ 0.05).Conclusions ADL of primary angle-closure glaucoma patients has varying degrees of dysfunction,and disease course,average monthly income,anxiety,depression and visual quality of life are closely related with it.

Objective:Human adipose-derived stem cells (hASCs)are a highly attractive source in bone tissue engineering.To generate a luciferase reporter system that could be used to quantitatively and rapidly examine osteogenic differentiation potential of human adipose-derived stem cells (hASCs ) in vitro,and eventually make it possible to monitor the osteogenic differentiation of transplanted cells in vi-vo.Methods:The genomic DNA harboring promotor regions of osteocalcin and DNA sequences encoding luciferase genes were amplified by PCR and cloned into the pLVX-pTRE-puro vector to generate the OCpro-Luc-Puro construct.Then,the OCpro-Luc-Puro construct together with three assistant vectors:pM-DLg/pRRE,pRSV-REV,and pVSVG,were transiently transfected into HEK293T cells followed by viral supernatants collection,filtration and concentration.Next,the hASCs stably expressing luciferase repor-ter gene driven by osteocalcin promotor were created with the lentivirus carrying OCpro-Luc-Puro cassette under puromycin selection.The OCpro-Luc-hASCs were then cultured in the absence or presence of osteo-genic differentiation medium.On the 7th and 1 4th days,after osteogenic induction,cellular extracts were collected and analyzed by luciferase reporter assay.Meanwhile,alizarin red staining and quantification as well as quantitative reverse transcription PCR (qRT-PCR)analysis of osteogenic associated genes osteo-calcin (OC),runt-related transcription factor 2 (Runx2)and alkaline phosphatase (ALP)were used to assess the osteogenic differentiation ability of OCpro-Luc-hASCs.Results:OCpro-Luc-Puro plasmid and OCpro-Luc-hASCs were successfully generated.On the 7th and 1 4th days after osteogenic induction,the luciferase activity of the cellular extracts from OCpro-Luc-hASCs was dramatically increased.Consistently, the extracellular matrix mineralization,as shown by Alizarin red S (ARS)staining and quantification was also markedly intensified and a marked increase of the mRNA expression levels of OC,Runx2 and ALP, although to variable extent,was observed upon osteogenic differentiation.These results indicated that the observations from traditional experiments examining hASCs osteogenic differentiation were largely in agreement with that of our luciferase reporter assay in OCpro-Luc-hASCs.Conclusion:We established a luciferase reporter system that could be used to rapidly,quantitatively and specifically determine osteo-genic differentiation ability of hASCs.Comparing with the traditional time-consuming methods,the system we generated here was highly effective.This system not only can be used to examine ostogenic differentia-tion of hASCs in a high throughput manner,but also provides a way to monitor ostogenic differentiation of cells in vivo.

Objective:To investigate the effect of human p27~(kip1) gene on the proliferation of human tongue squamous cell carcinoma(TSCC) Tca8113 cells.Methods:Human p27~(kip1) gene was cloned from normal tissue and reconstructed into eukaryotic expression vector pcDNA3, the recombinant plasmid was transfected into Tca8113 cells with liposome.The integration and expression of exogenous objective gene in the target cells were examined by PCR and flowcytometry,the growth of transfected cells and non-transfected cells was investigated by MTT assay and flowcytometry.Results:p27~(kip1) cDNA was successfully cloned and reconstructed into vector pcDNA3.The gene was trasfected into Tca8113 cells and stable transfection was achieved by screening with G418.After p27~ kip1 transfection the cell proliferation was inhibited,proliferation index decreased(P

Objective To explore the HLA-A2 restriction and immunogenicity of 5 previously identified HCV-speeific CTL epitopes. Methods Based on T2 cell, to explore the HLA-A2 restriction of previously identified HCV-specific CTL epitopes by MHC-peptide complex stabilization assay;To detect pep-tide-specific CTL in HLA-A2~+ PBMC stimulated by HLA-A2-restricted peptides by intracellular cytokine staining(ICS) and ELISPOT; To explore the cytotoxicity of peptide-specific CTL to same peptide-loaded T2 cells (target cells) by CTL cytotoxicity test. Results Among 5 previously identified CTL epitopes NS4b_78 (SMMAFSAAL) and NS5a_367 (TVSSALAEL) have high-affinity for HLA-A2 molecules(FI 1) ;ELISPOT results shown that NS4b_78(SMMAFSAAL) and NSSa_367(TVSSALAEL) induced high levels of IFN-γ-se-creting cells [(60±6) SFC/10~4 PBMC vs (4±1 ) SFC/10~4 PBMC, P < 0.01 ; (10 ± 3 ) SFC/10~4 PBMC vs (2±1 ) SFC/10~4 PBMC, P <0.01, respectively] ;ICS results indicated that there were high percentages of CD8~+ IFN-γ~+ T cells in total CD8~+T cells stimulated by these peptides [(2.33 ±0.22 ) % vs (0.05±0.01)%, P <0.001 ; (0.36±0.06)% vs (0.03±0.01)%, P <0.001, respectively]. Furthermore,peptide-specific CTL could effectively kill same peptide-loadcd T2 cells. Conclusion NS4b_78 (SMMAF-SAAL) and NSSa_367 (TVSSALAEL) were identified as HLA-A2-restricted CTL epitopes which could in-duce immune response in vitro.

ObjectiveTo investigate the effect of the displacement of the selected silver clip in the different respiratory state achieved by active breathing control ( ABC ) system on the displacement of the geometry constituted by all of the silver clips at the border of the cavity in external-beam partial breast irradiation (EB-PBI).MethodsTwo sets of CT images in state of moderate deep inspiratory breathing hold (mDIBH),deep expiratory breathing hold (DEBH),and free breath (FB) were acquired in the same CT simulation assisted by ABC system for each of the 27 patients after breast conservative surgery.All silver clips in the cavity were delineated based on each set of CT images.Thereafter,the irregular geometry based on the silver clips as the vertices was automatically formed.Four selected clips located at the top,bottom,lateral border and medial border of the cavity were correspondingly manually registered based on automatic registration of the CT images acquired in the same or different state of respiration including mDIBH,FB,and DEBH.The displacement of center of the geometry in the direction of right-left (RL),anterior-posterior (AP),and superior-inferior (SI) separately based on automatic registration and manual registration was evaluated.The difference of the displacement was analyzed by Kruskal-Wallis H-test and Kolmogorov-Smirnov Z-test.Results When registered between mDIBH and mDIBH,FB and FB,DEBH and DEBH,the differences of the displacement of the center of geometry were not statistically significant (H =0.00 - 1.76,P=0.184-0.954). When registered between mDIBH and DEBH,the differences were statistically significant ( Z =11.31 - 23.00,P =0.000 - 0.001 ).There were statistically significant differences in the displacement of geometry center based on the selected silver clip between different registration forms in AP and SI directions (Z=4.76-25.54,P=0.000-0.029).ConclusionsThe difference of intrafraction displacement of the geometry constituted by the clips between the same respiratory states in the three dimensional direction is not statistically significant,but the difference is statistically significant between the different respiratory states in AP and SI directions.