Objective To investigated the biological procedure of allograft decalcified bone matrix(DBM)and bone cement(BC)combined with bovine bone morphogenetic protein (bBMP)used for the repair of femoral defect caused by microwave－ induced hyperthermia in dogsby 99mTc－ MDP bone scintigraphy.Method The canine femoral defect(length 25mm,width 10mm)was caused by microwave－ induced hyperthermia(50℃ ,20minutes)and the composite material was implanted .Then the canine femurs were examined by 99mTc－ MDP bone scintigraphy respectively at different postoperative time and the results were compared with that of X－ ray photography and histological observation.Bone cement was implanted in the other femur as a contrast.Results It could be observed at the first and the second month that the radioisotope was gathered in the place where the composite material was implanted and the amount of radioisotope gathered in was the most abundant at the third month and it was lasted to the fourth month. That of the sixth month was decreased to that of the second month.The radiation count of the first, the second, the third the fourth and the sixth month were 93.9± 12.7, 110.7± 16.4,222.1± 24.0,201.3± 26.9 and 111.6± 20.7 respectively,and the count of the third month and the fourth month were more than that of the first, the second and the sixth month(P<0.01).Conclusion The composite material could be remodeled easily and the new bone could be formed by the induction of bBMP. So it could be merged with the normal bone.While the 99mTc－ MDP bone scintigraphy is the object and reliable index to determine the biological procedure of the composite material in dogs.

AIM To establish anti-osteosarcoma antibody producing hybridoma cell lines and to study the characterization of the monoclonal antibodies. Methods BALB/c mice were immunized with human osteosarcoma cells OS-9607 and the immunized spleen cells were fused with SP2/0 cells to raise hybridoma. The propert of antibody and it's cytotoxic effect were studied respectively with immunohistochemistry methods using OS-9607 and normal hepatocytes、 Western Blot methods and MTT method. Results A hybridoma cell line named 3D9 was established and it secreted high quality mAbs steadily. 3D9 cell had all the characteristics of hybridoma. The mAb's corresponding antigens was specifically and highly expressed in human osteosarcoma. With enzyme-labeled immunohistochemical staining on formaldehyde -fixed sections from human osteosarcoma,it was found that 83％ of the specimens expressed the corresponding antigen. Most of them were expressed on the nuclear of cells, no positive expression was observed in kinds of normal tissues. Western Blot showed 3D9's corresponding molecule weight is Mr54 000. MTT assay proved that the cytotoxicitis of effective groups were higher than control groups. Conclusion A high quality hybridoma is cultured and the mAb secreted by it has osteosarcoma specificity and obvious cytotoxic effect. It may be a new biochemical mark of osteosarcoma, and it's clinical prospect of immunotherapy will be wide.

Objectives:To study the effects of gene therapy with tissue type plasminogen activator(t PA)cDNA on the formation of thrombo embolism in vascular anastomotic sites. Methods:①The cDNA encoding t PA was amplified by RT PCR using the isolated total RNA as the template from the Bowes melanoma cells.②Recombinant plasmid pAdCMV t PA was cotransfected into 293 cells with pJMa 17 ,and the infectious but replication deficient AdCMV t PA was generated.③The rats were randomly divided into the control and treatment groups.11 0 nylone medical suture was applied to perform rat carotid artery end to end anastomoses.In the treatment group,AdCMV t PA solution was injected into the vascular anastomotic site while AdCMV (no containing t PA DNA) solution was injected into the control group. By means of RT PCR and chromogenic plasmin substrates,the following results were obtained. Results:①The t PA cDNA was successfully cloned and its eukaryotic expressing vector was constructed.②When the isolated RNA was performed with RT PCR,1.69 kb band appeared in the treatment group while the band could not be found in the control group.The t PA activity could be detected postoperatively on the 1st,2 nd,3 rd,4 th,5 th,6 th,7 th,10 th and 13 th day of the treatment,but could not be detected in the control group. Conclusions:The t PA gene can produce t PA having biological activity at anastomotic sites, possibly prevent the formation of thrombus embolism effectively and develop the anastomotic patency.

Objective To clarify the temperature curve of the irradiation target area,its adjacent tissue and the whole body during extracorpereal microwave irradiation, then to compare and optimize different irradiation models. Methods Different parts of the chest of adult New Zealand white rabbit were irradiated using different extracorporeal microwave irradiation models. The temperature of the irradiated skin, the subcutaneous and deep parts, the adjacent tissues and the anus was measured. The experiment was bi-factor and multi-level designed according to the repeatedly measured data and the rabbits was divided into group a,b,c and d. Results The increase rate of the surface temperature in the dorsal lung was similar between group d and group b1(F=10.04,P<0.01). However,the increase rate of the surface temperature in the ventral lung of group d was lower, and the mean temperature of this site measured 10 minutes later was also lower than group b1(F=10.04,P<0.01). The increase rate of the rectal temperature of group d was higher,and the mean rectal temperature tested 10 minutes later was also higher than group b1(F=7.04,P<0.01). Conclusions Multi-array irradiation could achieve satisfactory irradiation depth and appropriate therapeutic temperature. Well controlled extracorporeal microwave irradiation under is an ideal thermotherapy method.

Objective:To investigate the occurrence rate and distribution pattern of three-rooted mandibular molars in children using CBCT.Methods:CBCT images of 206 children aged 4-11 years were retrospectively analyzed.The root numbers of the bilateral man-dibular first and second deciduous molars,and the permanent mandibular first molars were recorded.A chi-square test was used to de-tect the gender and side difference.Spearman correlation test was used to analyze the relation between the bilateral homologous teeth and the mandibular first and second deciduous molars on each side.Results:The occurrence rate of three-rooted mandibular first and second deciduous molars and mandibular permanent first molars was 10.9％(19/175),25.1％(46/183)and 28.4％(57/201)for individuals,respectively(x2=18.543,P＜0.01),and 8.7％(32/366),19.0％(73/384)and 23.6％(95/402)for teeth(x2=30.692,P＜0.01),re-spectively.Gender difference was not detected for each tooth type(all P＞0.05),while side differences were detected in mandibular sec-ond deciduous molars and the frequency of three-rooted molar on the right and left side was 23.7％and 14.2％(P＜0.05),respectively.The concurrence rate of bilateral three-rooted mandibular deciduous first and second molars and mandibular permanent first molars was 57.9％(11/19),47.8％(22/46)and 66.7％(38/57),respectively,and the rho was 0.710,0.597 and 0.745,respectively(all P＜0.01);between the three-rooted deciduous first and second molars on the left side,rho=0.188(P＜0.05)and on the right side,rho=0.304(P＜0.01).Conclusion:The occurrence rate of three-rooted mandibular molars in children increases in the following sequence:first deciduous molars＜second deciduous molars＜first permanent molars.They frequently occur bilaterally and exhibit a moderate to high degree of correlation.However,the correlation was very weak between the mandibular deciduous first and second molars.

Objective To evaluate the prognostic factors of spinal metastases by recursive partitioning analysis (RPA)and establish a decision tree model that can guide clinicians to select individualized treatment.Methods Between January 2011 and December 2015,three institutional databases were searched to identify 169 patients with metastatic spinal tumors underwent surgery.The ratio of male and female was 1.48:1 (102 males and 67 females).The average age was 59.2±11.1 years.One-hundred eighteen cases of patients were randomly selected as training samples and the remaining 51 cases were verified samples.Preoperative factors were collected and analyzed by RPA methods,including primary tumor,KPS score,Frankel grade,gender,age,visceral metastasis,bone metastasis,spinal metastasis,blood glucose,blood pressure,surgery site,symptoms,surgery interval,serum albumin level and other risk factors.Pearson's Chi-square test was performed to judge the segmentation point of the decision tree model.The decision tree model is built using the Chi-square automatic interaction detector (CHIAD) algorithm and sensitivity and specificity was automatically calculated.Results The median postoperative survival time was 12.6±1.2 months [95％CI(10.1,15.0)].70％ were randomly selected as the experimental group (118 cases),and 30％ were the verification group (51 cases).The sensitivity was 96.9％.The specificity was 89.8％.The Kappa coefficient was 0.874 in the experimental group.The sensitivity was 95.4％.The specificity was 90.8％.The Kappa coefficient was 0.810 in the test group.The prognostic factors (weight from high to low) based on RPA were Frankel grade (F=8.132,P=0.005),the primary tumor and the KPS score (Equal,F=9.871,P=0.000 and F=1 1.945,P=0.003),serum albumin and movement time (Equal,F=7.566,P=0.018 and F=9.966,P=0.008).The decision tree model consists of 7 types.Survival time was 51 months,18 months,13 months,8 months,4 months,5 months,9 months in the class Ⅰ～Ⅶ,respectively.According to the difference of RPA survival time,the operation was classified as 3 grade.Total spinal resection was regarded as grade 1,including class Ⅰ and class Ⅱ,with an average survival time of over 18 months.Limited operation was regarded as grade 2,including class Ⅲ,Ⅴ and Ⅶ with survival time in 6-18 months.The conservative treatment was regarded as grade 3,including class Ⅳ and Ⅵ with the survival time was less than 6 months.Conclusion The decision tree model based on RPA for predicting the survival time of spinal metastases can not only identify the prognostic factors,but also classify and grade various prognostic factors;the decision tree model is simple and can guide clinicians to choose the best surgical plan by predicting the survival time.

OBJECTIVE To establish a high-performance liquid chromatography method for the simultaneous determination of 14 components including geniposide, mangiferin, baicalin, berberine hydrochloride, wogonoside, baicalein, aloe-emodin, rhein, wogonin, emodin, praeruptorin A, chrysophanol, physcion and praeruptorin B in Qingfei Yihuo tablets. METHODS Titank C18 (250 mm×4.6 mm, 5 μm) column was used; 0.1% phosphoric acid solution(A)-acetonitrile(B) was used as the mobile phase with gradient elution; detection wavelengths: geniposide at 238 nm, aloe-emodin, rhein, emodin, chrysophanol, physcion and mangiferin at 254 nm, berberine hydrochloride at 265 nm, baicalin, wogonoside, baicalein, and wogonin at 280 nm, praeruptorin A and praeruptorin B at 321 nm. The flow rate was 1.0 mL·min−1 and the column temperature was 35 ℃; the injection volume was 10 μL. RESULTS The linear ranges of geniposide, mangiferin, baicalin, berberine hydrochloride, wogonoside, baicalein,aloe-emodin, rhein, wogonin, emodin, praeruptorin A, chrysophanol, physcion and, praeruptorin B were 4.96−223.17, 0.84−42.22, 18.76−938.16, 4.46−223.17, 4.86−243.10, 1.59−79.32, 0.76−38.17, 1.03−51.49, 1.59−79.40, 1.21−60.72, 1.80−90.06, 0.91−45.48, 1.04−51.83 and 0.86−43.23 μg·mL−1, with r all ≥ 0.999 9. The RSDs of instrument precision, stability and reproducibility tests were <3%, and the average recoveries in sample(n=6) were >90%, with the RSDs <3%. CONCLUSION The method is simple and reproducible and can provide a scientific basis for improving the quality standard of Qingfei Yihuo tablets.