Objective To explore the changes of human mitochondrial COXI,ND1 and ND6 genes expression induced by ionizing irradiation.Methods Changes of human COXI,ND1 and ND6 gene expression were detected by RT-PCR and Real-time PCR 8 h after the irradiation in human lymphoblastoid cell lines,which were exposed to 1-10 Gy 60Co γ-rays.And the dose-effect relationships between expression changes of the genes and the doses were analyzed.The changes of these three genes expression were also analyzed at different post-radiation time-points between 0.5 h and 72 h after irradiation of 5 Gy in order to explore the time-effect.Results The expression of three genes COXI,ND1 and ND6,showed either the dose-effect or the time-effect after irradiation.The gene expression levels of three genes up-regulated generally and the peak change time-point was 4 h after irradiation.Conclusion Ionizing radiation,msht induce the changes of mitochondrial gene expression,and the gene expression level is up-regulated.

Objective To investigate the effect of mixed application of lecture-based learning and flipped classroom and problem-based learning in microbiology teaching. Methods The five-year program students of Grade 2013 and Grade 2014 were randomly divided into two groups, and the experimental group (48 students in the teaching reform class) carried out the blended teaching, while the contrast group (48 students of parallel class) carried on the traditional teaching. After the lecture, the teaching effect of the hybrid teaching method and traditional teaching method was analyzed after the theoretical and the experi-mental operation test were adopted, and the four part questionnaire surveys including the integrated use of knowledge, active classroom atmosphere, innovation ability and teaching satisfaction were proceeded in each group. The data of each group was analyzed by t test analysis with SPSS 19.0 respectively. Results The theory test scores of experimental group was (90.16±3.14), which was higher than the control group (82.33± 4.21). The difference between them was very significant (P=0.000). Survey results showed that the integrated use of knowledge, active classroom atmosphere, innovation ability and teaching satisfaction were higher than traditional group. The difference was statistically considered significant (P<0.01). Conclusion Hybrid teaching method is very good for training and development of students' comprehensive quality and ability. It has important significance in improving the students' score.

Objective:To construct GM-CSF and EB virus LMP2A fusion gene recombinant adenovirus and identificate it , using recombinant adenovirus to do immunologic study.Methods:With RT-PCR and Western blot to detect GC 2A gene expression of the recombinant adenovirus , with ELISA to analysis antibodies of vAd-GC2A stimulation in murine body , lactate dehydrogenase assay to detect the mouse cellular immunity effect , thereby the recombinant adenovirus immune effect was initially analysis.T-test method was used to do preliminary analysis and evaluation of the immune effect of recombinant adenovirus .Results:GC2A fragment amplified by PCR, the size was 1 961 bp and it was the expected.The results showed, the recombinant adenovirus vAd-GC2A proteins can be recognized by sera antibody of GM-CSF ( LMP2A); the results of ELISA showed that vAd-GC2A can stimulate the generation of antibodies.EBV-positive tumor cell killing rate (%) by CTL induced recombinant adenovirus was (66.7 ±6.9)%, at the same time the adenovirus type 5 wild strains and PBS were (24.3 ±2.5)%and(32.4 ±3.1)%only(P≤0.05).Conclusion:Fusion gene has been successfully constructed , vAd-GC2A in mice have humoral and cellular immune function.The data suggest that the vAd-GC2A should play a potent role in preventing the positive EB virus tumor.

Objective To identify the expression of a fusion gene GCA formed from GM-CSF gene and LMP2A gene of Epstein-Barr virus (EBV) in a recombinant BCG (rBCG) and to study its immunoge-nicity.Methods The rBCG was constructed to express the fusion gene GCA and the expressed products were detected by Western blot assay .ELISA was performed to measure specific antibody titers in serum sam-ples from mice immunized with rBCG .Lactate dehydrogenase assay was used to analyze the cellular immuni-ty of mice.A mouse model of EBV-positive gastric carcinoma was established to evaluate the therapeutic effects of rBCG.Results The target proteins of GM-CSF and LMP2A were successfully expressed in rBCG . The specific antibodies were detected in rBCG immunized mice as indicated by ELISA .The maximum anti-body titer reached 1 ∶27 900 [(326.5±7.8) pg/ml] as injection with rBCG 5×108/mouse.The rBCG in-duced cytotoxicity of cytotoxic lymphocytes (CTLs) to EBV-positive gastric carcinoma cells (GT39) (with a killing rate of 89.6%±6.8%) was significantly higher than that of control group (P<0.05) The sizes of tumor in PBS control group [(1964.0±548.7) mm3] and BCG group [(1268.65±72.4) mm3] were big-ger than those in rBCG group [(168.64±78.80) mm3].Conclusion The rBCG expressing GM-CSF and LMP2A fusion gene was successfully constructed .The rBCG could induce humoral and cellular immune re-sponses in mice and inhibit the growth of tumor .

Objective To establish the analysis criteria of nucleoplasmic bridges (NPB) and a dose-response curve of NPB frequencies in human peripheral blood lymphocytes irradiated by 60Co γ-rays.Methods Human peripheral blood samples were collected from three healthy males,and were irradiated with 0,1,2,3,4,5 and 6 Gy of 60Co γ-rays.A cytokinesis-block micronucleus assay was carried out to analyze NPBs and micronuclei (MN) in binucleated cells.Results NPBs in binucleated cells at each dose level of γ-ray was conformed to the Poisson distribution.The dose-response curve of the γ-ray-induced NPB frequencies in human peripheral lymphocytes followed the linear-quadratic model y =(1.39 × 10-3)x2 + (4.94 × 10-3)x (R2 =0.981,P ＜ 0.05).Conclusions The dose-response curve of NPB frequencies in human peripheral blood lymphocytes induced by 0-6 Gy 60Co γ-rays was established.

Objective: To develop an HPLC method for the determination of rutin, hyperin and quercetin in Vicia sepium L. . Methods:The samples were separated on an Agilent ZORBAX Eclipse XDB-C18 column(250 mm × 4. 6 mm, 5 μm) with the mobile phase consisting of acetonitrile-1‰ phosphoric acid solution with gradient elution at the flow rate of 0. 8 ml·min-1 . The column tem-perature was 30℃, and the detection wavelength was set at 370 nm. Results: The linear range of the three components was 4. 090-130.940 μg ·ml-1(r=0.999 9), 4.600-147.200 μg ·ml-1(r=0.999 9) and 0.810-25.780 μg·ml-1(r=0.999 8), and the average recovery was 103. 45% (RSD=1. 25%), 98. 96% (RSD=1. 77%) and 102. 88% (RSD=0. 84%)(n=6), respectively. Conclusion:The method is stable, reproducible and simple, which can be used in the quality control of Vicia sepium L. .

BACKGROUND:Recombinant adeno-associated virus serotype 9 has a high affinity in myocardial tissue, and the expression of recombinant adeno-associated virus serotype 9-enhanced green fluorescent protein (rAAV9-eGFP) in the aorta of atherosclerosis mice is not clear. OBJECTIVE:To explore the optimal time point of rAAV9-eGFP expression in the aorta of atherosclerosis mice. METHODS:Atherosclerosis model was established with high-fat diet in 30 ApoE-/-mice for 16 weeks. Among them, 25 mice were injected with 5.0×1011 vg (virus genomes) rAAV9-eGFP through the tail vein, while the remaining 5 mice were injected with saline, serving as the control group. The virus-transfected mice were kil ed at 14, 21, 28, 35 and 60 days after transfection, and aortic tissue was harvested. The expression of enhanced green fluorescent protein was detected with laser scanning confocal microscope. Western blot assays were used to detect the expression of enhanced green fluorescent protein in aorta. The expression of enhanced green fluorescent protein in vivo was observed and the optimal expression time point was determined. RESULTS AND CONCLUSION:rAAV9-eGFP effectively transfected the aorta of atherosclerosis mice, enhanced green fluorescent protein was expressed in aortic tissue, and the expression intensity increased gradual y with the increasing transfection time. The highest expression level was found at 35 days after transfection and then maintained stable at 60 days. There were significant differences at different time points after transfection (P<0.001). These data indicate that rAAV9-eGFP can be effectively expressed in the aorta of atherosclerosis ApoE-/-mice and rAAV9-eGFP can be regarded as the optimal vector in the treatment of atherosclerosis.

Objective To investigate the nutritional risks, undernutrition, and nutritional support among inpatients in tertiary hospitals in Tianjin using Nutritional Risk Screening 2002 (NRS2002). Methods Inpatients in six departments in two tertiary hospitals in Tianjin (Tianjin Nankai Hospital and Tianjin Chest Hospital ) were consecutively enrolled from March 2005 to March 2006. Their nutritional risks were screened using NRS 2002, and the nutritional support was investigated. Results A total of 1200 inpatients received nutritional screening, and 93.0% of them underwent NRS 2002 scoring. The prevalence of undernutrition was 9. 8% and the prevalence of nutritional risk was 42. 8%. Of these patients, 241 patients (46.4%) with NRS2002≥3 received nutrition support, and 244 patients (35.9%) with NRS2002 ＜3 received nutritional support Conclusions NRS2002 is suitable for nutritional risk screening among inpatients. Inpatients usually have nutritional risks or undernutrition. However, physicians at different levels in different regions have different awareness of such risks or conditions, and the clinical application of nutritional supports sometimes are inappropriate.

Molecularly imprinted polymers (MIPs) with high selectivity to tilmicosin (TIM) were prepared using tylosin(TYL) as dummy template, methacrylic acid(MAA) as monomer, ethylene glycol dimethacrylate (EGDMA) as cross-linker.The effects of 4 porogens including dimethyl formamide, methanol, acetone, and chloroform on the recognition capability of MIPs were investigated.Orthogonal test was used to optimize the preparation of MIPs, and the optimal composition was as follows; 1.0 mmol TYL, 8.0 mmol MAA, 20.0 mmol EGDMA, 6.0 mL chloroform, 20.0 mg azobisisobutyronitrile.The solid phase extraction condi tions and characteristics of MIPs as adsorptive material for the selective extraction and enrichment of TIM were also studied.The recovery of TIM was above 90% when the following procedure was applied to MIPs cartridge: conditioning with methanol and water(pH 9.0), loading with acetonitrile, cleaning with methanol and chloroform respectively, and eluting with 3 mL methanol-ammonia(95:5, V/V).The recovery of TIM on non-imprinted polymers cartridge was only 32%.

Objective To explore if the occupational exposure to low dose thorium could induce adaptive response in peripheral lymphocytes.Methods 40 individuals.who exposed to thorium and rare earth mixed dust(exposure group) or control in Baotou Steel Plant, were selected, and chromosome aberrations were analyzed.Then the peripheral blood samples were irradiated in vitro with 2 Gy60Co γ-rays,and unstable chromosome aberration or DNA stand breakage analysis using single cell gel electrophoresis was performed. Results The dicentrics before 2 Gy exposure in exposure group was higher than that in control(P>0.05). But the dicentries after 2 Gy exposure in exposure group was lower than that in control,but not significantly (P>0.05).The tricentrics in exposure group was significantly lower than that in control(U=3.1622, 0.0ol