Objective:To observe the proliferation and the free intracellular calcium ion concentration of osteoblast under the effect of static magnetic field and discuss the possible mechanism of the static magnetic field on promoted the proliferative function. Methods: The MTT assay was used to measure the rate of the proliferation on different magnetic field intensity(0, 0.026, 0.044, 0.090 T)and action time(24, 48, 72 h).The fluorescence double wavelength spectrophotometer was used to measure the concentration of intracellular calcium ion. The data obtained were analyzed for ANOVA and t-test using SPSS 11.0 software package. Results: Osteoblasts in group 0.044 T appeared significant proliferation within 48 h(P

0.05) in GCF. Conclusion:, The change PGE 2 and ALP in GCF of periodontitis tooth was similar to that of normal tooth during orthodontic therapy.

Objective:To Study the effect of static magnetic field on the proliferation and differentiation of osteoblasts.Methods:In vitro cultured rat calvarial osteoblasts were seeded into 96-well culture plate and exposed to the different magnetic intensity field for 24,48 and 72 h respectively.MTT assay was applied to study the cell proliferation. ALP was measured by a microplate reader.Results:When the magnetic exposure time extended to 48 h or 72 h, the corresponding MTT value of osteoblasts with magnetic treatment of 40 mT or 62 mT increased obviously(P0.05). ALP activity increased after 24 h exposure to static magnetic field, the effect of static magnetic field intensity of 62 mT and 83 mT showed the strongest promotion effect(P

0.05). But there was more BMP in the treated group of periodontitis than the other groups(P

Objective To analyze and compare the accuracy and utility of determining sperm DNA integrity by sperm chromatin dispersion (SCD) test and acridine orange staining (AO) test.Methods The level of DNA fragmentation was determined by SCD test and AO test in 32 adult healthy fertile men and 27 idiopathic oligozoospermia (IO) patients.Sperm nuclei with large DNA dispersion halos or with mediumsized halos were normal and nuclei with small-sized halos or no halo were abnormal.The normal sperm DNA was double strands and stained by AO as green.The damaged sperm DNA was single strand,which were stained as green for native DNA and red for denatured DNA.Results The percentage of sperm nuclei with large halos,medium-sized halos,small-sized halos and no halo evaluated bv SCD test in IO patients were (49.9±13.8)%,(11.5±5.4)%,(11.9±6.1)%and(26.7±10.0)%,and they were(73.2±6.2)%,(14.7±6.3)%,(6.8±2.9)%and(5.3±2.2)%in healthy control group,respectively.There was significant difference between two groups(t=8.576,P＜0.01;t=2.083,P＜0.05;t=4.284,P＜0.01;t=11.823,P＜0.01).The percentage of sperm DNA fragmentation was (38.6±12.1)%in IO patients and was (12.1±5.2)% in fertile men,respectively.A statistically significant difference was found between IO patients and healthy control group under SCD test (t=11.995,P＜0.01).AO test showed no significant differences between IO patients and healthy control group (t=1.626,P＞0.05).The percentage of sperm DNA fragmentation evaluated by AO test in IO patients was (45.5 ±13.8)%,and it was (39.8±13.3)%in healthy control group.Conclusions Sperm DNA fragmentation may lead to male infertility.The SCD is effective in testing the sperm DNA fragmentation as a screening procedure to determine semen quality during basic infertility investigation for clinical use.

Objective To study AngⅡ induced apoptosis of HUVECs and to observe the protective effect of serum contained huangqi on endothelial cell.Methods ECV-304 cells were randomly divided into control group,AngⅡgroup and huangqi group.In the control group,cells were cultured for 18 h,and the concentration of AngⅡ were 0 mol/L,1×10-6 mol/L,1×10-5 mol/L and 1×10-4 mol/L.The cell proliferation was measured by MTT assay.Electron microscope was used to observe the change of HU-VECs.Ultrastructure of HUVECs induced by AngⅡ was observed by electron microscope.In the huangqi group,serum contained huangqi of different concentration were added into the medium and cultured for 24 h,then AngⅡ of 1×10-4 mol/L was included and cultured for 18 h,and the apoptosis ratio induced by AngⅡwas measured by flow cytometry.Results AngⅡof different con-centration could all significantly inhibit HUVECs proliferation.AngⅡof different concentration could all induce endothelial cell ap-optosis.HUVECs apoptosis was observed by electron microscope.HUVECs apoptosis induced by AngⅡcould be inhibited by ser-um contained huangqi.Conclusion Serum contained huangqi could protect endothelial cells.

Objective:To investigate the effects of Porhyromonas endodontalis(P.e)liopolysaccharide(LPS)on the expression of IL-23mRNA and protein in mouse osteoblasts MC3T3-E1 and the role of phosphatidylinositol 3-Kinases (PI3K)signaling pathway in this process.Methods:MC3T3-E1 cells were treated with different concentrations of P.e LPS for different hours,or pretreated with LY294002,a special PI3K inhibitor.The IL-23 mRNA and protein expression levels were detected by real-time PCR and Western blot respectively.Results:The level of IL-23 mRNA increased in MC3T3-E1 cells with the increase of concentration and the treatment time of P.e LPS(P <0.05).The IL-23 protein expression was increased by P.e LPS in a time dependent manner(P <0.05).The mRNA and protein of IL-23 decreased(P <0.05)after pretreatment with LY294002.Conclusion:P.e LPS can induce the expression of IL-23 mRNA and protein in MC3T3-E1 cells,and the PI3K signaling pathway may play a part in this process.

Objective:To construct a esophageal cancer module with Chinese characteristics based on MD Anderson Symptom Inventory (MDASI) public scale, develop the Perioperative Esophageal Cancer Symptoms Assessment Scale combining above two parts.Methods:The original item pool was formulated through literature review, clinical interviews, and reference of existing symptoms assessment tools. After two rounds of expert evaluation and pilot survey, the preliminary Perioperative Esophageal Cancer Symptoms Assessmment Scale was developed combining Chinese MDASI (MDASI-C). A total of 150 perioperative esophageal cancer patients was assessed using the new scale, the included items were analyzed one by one, the reliability, validity and sensitivity of scale were checked.Results:Feasibility: the scale recovery was 100%, the completion rate of scale was 93.75%, the average completion time was 10 min. Reliability: the value of Cronbach α of the esophageal cancer module, MDASI-C, the combined scale were 0.747, 0.894, 0.883, respectively. Validity: the range of content validity index of items was 0.83-1.00, the scale-level content validity index average value was 0.93. Two common factors, which explained for 67.994% of variance, were extracted by exploratory factor analysis, the validity of criterion had statistical significance ( P<0.05). Sensitivity: the scores of the esophageal cancer module were significantly different among perioperative esophageal cancer patients with different Eastern Cooperative Oncology Group performance status ( H value was 9.264, P<0.05). Conclusions:The Perioperative Esophageal Cancer Symptoms Assessment Scale has good feasibility, reliability, validity and sensitivity, it is suitable for symptoms assessment of Chinese perioperative esophageal cancer patients.

Objective To observe the changes of expressions of brain-derived neurotrophic factor (BDNF)and its receptor tyrosine kinase B(TrkB)and in rats with hemitransectional spinal cord injury(SCI)after electroacupuncture on Du Meridian and rehabilitation training.Methods The animal model of acute hemitransectional lesion at the right half of T11 spinal cord was established in 96 adult female rats,which were then divided randomly into an electroacupuncture group,a rehabilitation training group,an electroacupuncture combined with rehabilitation training group and a control group.All the groups received treatment on the 3rd d after operation.The electroacupuncture group and rehabilitation training group were given electroacupuncture on points of Du Meridian and rehabilitation training,respectively,and the combined group was given Du Meridian electroacupuncture in addition to rehabilitation training.Basso-Beattie-Bresnahan(BBB)scale was used to evaluate motor function every week.Twelve rats of each group were sacrificed 4 and 8 weeks after operation,respectively,and their spinal cord tissues were extracted.The polymerase chain reaction(PCR),reverse transcription-polymerase chain reaction(RT-PCR)and immunohistochemical techniques were used to detect the expressions of BDNF and TrkB.Results BBB grade increased gradually as time went on.There were significant differences between control group and other groups at the same time point(P ＜ 0.05).The scores increased obviously in electroacupuncture combined with rehabilitation training group compared with electroacupuncture group and rehabilitation training group(P ＜ 0.05).The result of immunohistochemical observation and RT-PCR also showed that there were significant differences of expressions of BDNF and TrkB among control group and other groups at the same time(P ＜ 0.05).The effects in electroacupuncture combined with rehabilitation training group were much more obvious than those in electroacupuncture group and rehabilitation training group(P ＜ 0.05),but there was no significant difference between electroacupuncture group and rehabilitation training group (P ＞ 0.05).Conclusions Electroacupuncture on Du Meridian combined with rehabilitation training had synergic effect on rat's SCI,and could obviously improve the restoration of rat's motor function; the mechanism maybe related to the upregulation of expressions of BDNF and TrkB.

Objective To construct a prokaryotic expression vector pET-22b+with Middle East respiratory syndrome ( MERS) coronavirus nuclocapsid protein( NP) gene and to express and purify N protein.Methods N gene amplified by PCR was inserted into the prokaryotic expression vector pET-22b+.Recombinant plasmid was confirmed using DNA elec-trophoresis and sequencing.NP was expressed in E.coli BL21(DE3) by IPTG induced and purified by cation exchange chromatography using the AKTA purification system.Results The NP gene sequence was proved to be correct by sequen-cing and the protein was expressed in both soluble and insoluble forms in E.coli BL21 ( DE3 ) after IPTG induction.The purity and concentration of recombinant protein was improved obviously by cation exchange chromatography and enrich-ment.Conclusion Recombiant NP of high purity and concentration is purified and will facilitate NP functional research.