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 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 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 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 clinical effect of McDonald cervical cerclage and the affecting factors. Methods Between January 2002 to December 2013 in Peking University First Hospital we performed McDonald cervical cerclage for 116 single pregnant women. They were defined as the successful group who deliveried the live babies after 28 weeks after the cerclage and the failure group who deliveried in the second trimester. According to the surgical indications they were divided into preventive cerclage group and therapeutic cerclage group. Then we analyzed the curative effect and the affecting factors in the groups. Results (1) In the 116 cases, 12 cases (10.3%) failed, and 104 cases (89.7%) succeeded. In the successful group, 37 cases (35.6%,37/104) deliveried pretermly and 67 cases (64.4%) deliveried termly. And there were 56 cases of vaginal delivery (53.8%), and 48 cases (46.2%) of cesarean section. (2) Among the 116 cases, 48 cases (41.4%) were included in prophylactic cerclage group, the gestational age was (16.3± 2.2) weeks, 68 (58.6%) cases were included in therapeutic group, the gestational age was (24.0±2.2) weeks. The operation time was (22±9) minutes in preventive group and (24±13) minutes in therapeutic group,there was no statistical difference between the two groups (P>0.05). Live-birth rate between preventive cerclage group and therapeutic cerclage group was no statistically significant difference (P>0.05). The term birth rate (72.9%, 35/48) in preventive group was higher than that in therapeutic group (47.1%, 32/68), the difference was statistically significant (P<0.01). Neonatal hospitalization rate was lower in preventive group (14.6%, 7/48) than therapeutic group (36.8%, 25/68) , the difference was statistically significant (P< 0.01). (3) In the failure group placental pathology was examed in 7 cases. The placental tissue showed a large number of neutrophils infiltrating in 6 cases (6/7). In the successful group, 27 pregnant women deliveried between 28 to 33+6 weeks (26.0%,27/104), 10 pregnant women deliveried between 34 to 36+6 weeks 10 cases (9.6%, 10/104), 67 cases deliveried after 37 weeks (64.4%, 67/104). A lot of factors including maternal age, the previous cervix operation history, perioperative application of progesterone, operation time and preoperative invasive procedure were compared between the successful group and the failure group. Only maternal age and preoperative invasive proedcure were statistically significant (P<0.05) and the others had no statistical significance (P>0.05). (4) There were 68 cases in the therapeutic group, 7 cases failed, and 61 cases succeeded;the preoperative cervical os in failure group [ (21 ± 20) mm] was wider than that in successful group [(14±5) mm], the difference was statistically significant (P<0.05);and preoperative vaginal ultrasound measurement of cervical canal length were (18 ± 8) mm versus (19 ± 10) mm, there was no statistically significant difference (P>0.05). Conclusions The McDonald cervical cerclage for cervical incompetence is a simple, safe and high successful rate of intervention measures. The term labor rate of prophylactic cervical cerclage was higher than that of the therapeutic cerclage. Older maternal age and preoperative invasive procedure may be the risk factors for cerclage. The infection may play an important factor leading to the failure of McDonald cervical cerclage.

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.

ObjectiveTo explore the ultrasound characteristics of coronary artery fistula to improve its prenatal diagnosis.MethodsImaging data for 12 fetuses with coronary artery fistula who underwent fetal echocardiography at our hospital from June 2005 to December 2013 were retrospectively analyzed to summarize the ultrasound characteristics of coronary artery fistula.ResultsOf the 12 fetuses, 7 were confirmed with coronary artery fistula by echocardiography after birth, and the remaining 5 were aborted due to other cardiac abnormalities (since an autopsy was not performed, the diagnosis of coronary artery fistula could not be confirmed). Of the 7 confirmed cases, 1 had left coronary artery-to-right atrium fistula, 3 had left coronary artery-to-right ventricle fistula, and 3 had right coronary artery-to-right ventricle fistula; 1 had left coronary artery-to-right ventricle fistula with muscular ventricular septal defect, and 6 had simple coronary fistula. Two-dimensional ultrasound examination revealed that coronary arteries were displayed in all cases, and the diameter ranged from 1.8 to 4.0 mm. Color Doppler ultrasound clearly showed the fistula size, course and the situation fistulas drained into heart chambers. Diastolic regurgitation signal could be detected in the ascending aorta and aortic arch.ConclusionCoronary artery fistula can be diagnosed by color Doppler ultrasound combined with spectral Doppler ultrasound when the fetal coronary artery expands.

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.