Objective To evaluate the toxicity of the intra-articular injection of bevacizumab in the knee of the rabbit.Methods Thirty-two rabbits were divided into 3 experimental groups and normal control group.Three experimental groups were received intra-articular injection of bevacizumab (1, 2, 4 mg respectively) once every three weeks for two times and the normal control group was received the same amount of 0.9% sodium chloride solution.The animals were sacrificed after 6 weeks.Blood test was examined before and after treatment.Pathologic examinations of liver, kidney and artiluar tissue were taken after the sacrifice.The hematoxylin and eosin stain for synovium and cartilage were performed.The AB-PAS stain and Mankin's scale for cartilage were performed.Results All the rabbits kept normal physiological activity.There was no significant difference of major organs and articular tissue between experimental groups and normal control group.There was no significant difference for WBC, RBC, PLT, ALT, BUN and Mankin's scale among all groups.Conclusion No systemic toxicity effects were found for the intra-articular injection of bevacizumab in the knee of the rabbit.

Objective To evaluate the value of real-time three-dimensional echocardiography（RT3DE）to predict the long-term response to cardiac resynchronization therapy（CRT）.Methods Twenty-six patients with heart failure were scheduled for CRT.RT3DE was performed before and one year after the pacemaker implantation,and left ventricular（LV）dyssynchrony was defined as the maximum difference（Tmsv 16-Dif）and standard deviation（Tmsv 16-SD）when the minimum systolic volume of 16 segments was reached.Patients were divided into re-sponders and non-responders according to a reduction ≥15 % in LV end-systolic volume（LVESV）after CRT.Results Seventeen patients（65.4 %）were classified as responders.Compared with non-responders,the responders demonstrated a significant increase in LV ejection fraction,and reduction in Tmsv 16-SD.Tmsv 16-SD was a determinant factor for Δ LVESV ≥15%.Tmsv 16-SD 〉3.5% could be employed to evaluate the short-term response to CRT with 80 % sensitivity and 77 % specificity.Conclusions CRT can increase the LV systolic function and synchrony.RT3DE is highly predictive for long-term response to CRT

Objective To investigate the changes and clinical significance of serum total and high molecular weight adiponectin (HMW APN) levels in type 2 diabetic patients with non-alcoholic fatty liver disease (NAFLD).Methods Serum levels of total adiponectin and HMW APN were measured by ELISA,in 58 type 2 diabetic patients with nonalcoholic fatty liver disease (T2DM with NAFLD group),59 type 2 diabetic patients without nonalcoholic fatty liver disease (T2DM group),and 55 control subjects with normal glucose tolerance and without nonalcoholic fatty liver disease (NC group).Results (1) Alanine transaminase and total cholesterol levels were significantly higher in T2DM with NAFLD group than those in NC group(P＜0.01),while body mass index (BMI),aspartate aminotransferase,triglyceride (TG),and fasting insulin were also significantly increased (P＜0.05 or P＜ 0.01),and high density lipoprotein-cholesterol (HDL-C) were significantly decreased compared with those in NC and T2DM groups (P＜0.01).(2) Total adiponectin,HMW APN,and the ratio of HMW APN to total adiponectin in T2DM group were lower than those in NC group.Total adiponectin and HMW APN levels in T2DM with NAFLD group were significantly lower than those in T2DM group(P＜0.01).(3) Regression analysis showed that homeostasis model assessment insulin resistance index (HOMA-IR),TG,and HDL-C levels were independent predicting factors for total adiponectin and HMW APN levels.TG and BMI were independent risk factors for T2DM complicated with NAFLD,while total adiponectin and HMW APN levels were the protective ones (OR =0.701,0.489,respectively).Conclusions Hypoadiponectinemia may partially play an important role in the development and progression of NAFLD in T2DM.

To explore the regularity for the multiplication of avian influenza virus subtype H9N2 in large-scale microcarrier-based MDCK cell culture system, and to determine the optimal proliferation conditions. H9N2 subtype of avian influenza virus was inoculated into the MDCK cell growing on 24 well plate, and the HA titers of virus at different time were detected in the conditions of different infectious doses,different concentrations of TPCK- trypsin and different pH. The optimal conditions were determined. Then the H9N2 subtype avian influenza virus was grown in microcarrier-based MDCK cell in 250mL and 5L roller bottles. It was demonstrated that high viruse yield with a hemagglutination unit of 9 log2(1:512) could be obtained under the optimal conditions of multiplication . The result indicated the H9N2 subtype avian influenza virus could be produced in microcarrier-based MDCK cell in a large-scale culture system with a high virus yield and demonstrates the feasibility of the development of mammalian cell-based in influenza vaccine in microcarrier culture systems.

Objective To clone and express the hexon protein of three prevalent human adenovi -rus strains causing respiratory disease and analyze the antigenic characteristics of the recombinant proteins . Methods The full length genes encoding hexon protein of human adenovirus serotype 3(HAdV3), serotype 4(HAdV4) and serotype 7(HAdV7) were cloned by PCR and sequenced , respectively.The alignment anal-ysis was performed by using hexon gene sequences from GenBank .The major antigenic regions of hexon pro-tein of the three serotypes were expressed in E.coli and purified.The antigenicity, immunogenicity and cross reactivity of the recombinant proteins were determined by ELISA and Western blot assay .Results The full length gene sequence encoding hexon protein of human adenovirus serotype 4 was firstly reported in China , which showed more than 99%homology in both nucleotide and amino acid sequences with the human adeno-virus type 4 NHRC3 strain.The partial hexon protein sequence of HAdV 3, HAdV4 and HAdV7 containing all of the 7 hyperviriable regions ( HVRs) were expressed in E.coli, respectively .The purified recombinant proteins could be recognized by antiserum of the three serotypes of adenovirus .The antiserum samples against the three recombinant proteins could cross-react with particles of the three serotypes of adenovirus . The possible type-and species-specific epitopes were predicted .Conclusion The major antigenic regions of hexon protein of the three serotypes were successfully expressed .The purified recombinant proteins contai-ning both intertypes and type-specific epitopes showed a strong immunogenicity .

Objective To study the effects of endoplasmic reticulum stress on liver damage in young rats fed with high-fat diet.Methods We divided 48 male weaned young rats randomly into high-fat diet group and control group,which were separately fed with high-fat diet and normal diet.After feeding 8,12 and 1 6 weeks,the body weight and visceral fat of the rats were measured.The serum liver function was measured.The morphology of livers was observed by HE and transmission electron microscopy. The mRNA expressions of ATF6 and GRP78 in hepatocytes were measured with RT-PCR.Results ① The body weight and visceral fat weight of rats in high-fat diet group increased compared with those in control group (P <0.05).② Alanine aminotransferase and aspartate aminotransferase in high-fat diet group increased slightly over time (P >0.05);alanine aminotransferase at week 1 6 was increased significantly compared with that in controls (P < 0.05 ).③ Liver cells in high-fat diet group had steatosis at week 8 and the steatosis became more serious between week 12 and week 1 6.④ In high-fat diet group at week 8 there were a large number of lipid droplets in the cytoplasm,and the cell structure was close to that of normal cells;rough endoplasmic reticulum was nearly normal and the ribosome was visible.At week 12 and week 1 6,besides a large number of lipid droplets,we could also see that some substances with line-like structure deposited in rough endoplasmic reticulum pool.⑤ The expressions of ATF6 and GRP78 mRNA in hepatocytes in high-fat group at weeks 8, 12 and 1 6 were significantly increased compared with those in control group (P <0.05). Conclusion High-fat diet in infants can cause visceral fat accumulation,fatty degeneration of hepatocytes and liver injury.ATF6-mediated endoplasmic reticulum may be closely related to the liver injury which results from highfat diet.

Objective To establish a tree shrew mode of breast tumor.Method Forty-five 3 to 4 month-old female tree shrews were orally gavaged with 20 mg 7,12-dimethylbenz(a)anthracene (DMBA) or peanut oil per animal for three times.Following that, fifteen DMBA administrated tree shrews were implanted 90 day-release medroxyprogesterone acetate ( MPA) pellets.The tree shrews were palpated once weekly to detect mammary tumors for 45 weeks after first DMBA administration.Results DMBA were able to induce breast tumors (12.5%) in tree shrews, and MPA increased the tumor incidence (50%) while no breast tumors were observed in the control group.Three induced breast tumors were intraductal papillary carcinomas and one was IDC by H&E stain.Conclusion All induced tumors are similar with spontaneous tumors in structure and molecular markers.

Objective:To evaluate the effects of intraarticular injection of bevacizumab、sodium hyalu-ronate (SH)and 0.9% sodium chloride injection in the treatment of osteoarthritis (OA)in a rabbit model.Methods:Twenty-four male rabbits were randomly divided into bevacizumab group,SH group and control group after the model of OA had been made.The bevacizumab group and control group received intraarticular bevacizumab (4 mg)and 0.9% saline injection respectively once per three weeks for 2 times.The SH group received intraarticular SH once a week for 6 weeks.After 6 weeks,the histological examinations of cartilage and synovium,electron microscopy and expression of vasculan endothelial growth factorl (VEGF),for the synovium,expression of MMP-1 ,Mankin’s scale,macroscopic observation for cartilage were performed.Results:The histological observation of the bevacizumab group and the SH group showed that bevacizumab could decrease the synoviocytes and inhibit fibrous hyperplasia in synovial underlayer compard with the control group.Reduced apoptosis of chondrocytes and more integrated struc-ture of matrix and more glycosaminoglycan were also found in the bevacizumab group and the SH group compared with control group.The expression of VEGF and MMP-1 ,Mankin’s scale,macroscopic obser-vation were significantly decreased in the bevacizumab group compared with the SH group and the control group (P<0.05).Conclusion:Intraarticular injection of bevacizumab and SH can relieve inflammation of OA and alleviate the pathologic process of OA.The Bevacizumab was better than the SH in therapeutic effect,which maybe implicate a better choice for the treatment of OA.

Objective To investigate the effect of cerebrospinal lfuid replacement combined with vancomycin and dexamethasone intrathecal therapy on biochemical indicators of postoperative intracranial infection, in order to improve the clinical diagnosis and treatment. Methods 70 cases with intracranial infection collected in Third Hospital of Beijing Armed Police Corps from February 2010 to April 2013 were as subject, and randomly divided into two groups. Control group(n=35) were given cerebrospinal lfuid replacement and ceftriaxone intravenously, observation group(n=35) were given cerebrospinal lfuid replacement combined with vancomycin and dexamethasone intrathecal injection. The clinical effects and biochemical indicators were observed after treatment in two groups. Results In control group, the cure rate was 22.86%and total efifciency was 77.14%. In observation group, the cure rate was 37.14% and total efficiency was 91.43%. The differences between two groups were statistically significant (P<0.05). The differences of leukocytes, glucose, protein, intracranial pressure in two groups after treatment were also statistically signiifcant(P<0.05). Conclusion Cerebrospinal lfuid replacement combined with vancomycin and dexamethasone intrathecal injection therapy can increase intracranial infection.

Objective To investigate the anti-proliferation effect of peroxisome proliferator-activated receptor γ(PPARγ)agonist troglitazone(TGZ)on leukemic Raji cells and its mechanisms.Methods Raji cells in culture medium in vitro were given different concentrations of TGZ(0～60 μmol/L)for 24,48 and 72 h.The inhibitory rates of the cells were measured by MTT assay,cell apoptotic rate was detected by flow cytometry(FCM),agarose gel electrophoresis was used to observe the DNA ladder,and western blotting was used to analyzed the variation of apoptosis related proteins bcl-2,Bax and Survivin.Results TGZ(over 20 μmol/L)could inhibit the growth of Raji cells and cause apoptosis remarkably,the suppression was both in time-and dose-dependent manner.DNA ladder was observed after the cells treated by TGZ for 72 h,and western blotting analysis revealed that anti-apoptotie proteins Survivin and bcl-2 were decreased remarkably while pro-apoptotic protein Bax increased significantly after the cells were treated by TGZ for 48 h.Conclusion PPARγ agonist TGZ can inhibit the growth and induce apoptosis on Raji cells significantly,downregnlating the expression of Survivin and bcl-2 as well as upregulating of Bax expression of Raji cells may be one of its most important mechanisms.