ObjectiveTo study the correlation with Genes Coding forESBLs and ClassⅠIntegron in ESBLs-producing Escherichia coli from children.MethodsPCR was used for gene typing detection of 100 strains of ESBLs-producingEsche-richia colistrains. While detection of class I integrase gene in the 100 strains ESBLs-producingEscherichia coli and 100 strains of non-ESBLs producingEscherichia coli were separately performed by PCR. It provides the solid base not only to reveal the relationship between class I integron and drug resistance, but also the relationship between class I integron and ESBLs-producing. ResultsThe most frequently genotyping from ESBLs-producingEscherichia coli in children isCTX-M (84%), followed by TEM-1(63%). The predominant distribution of genotype in ESBL- producing strains isTEM-1 +CTX-M (45%), followed by CTX-M (34%). Class I integrase gene detected in ESBLs- producing and non- ESBLs producing strain were 100 cases (100%) and 25 cases (25%) separately, the difference was statistically signiifcant (P<0.05); drug resistance in class I integron positive strains were signiifcantly higher than in class I integron negative strains, especially in Ciprolfoxacin, Levolfoxacin, and Amino-glycoside (86.4%, 88%, and 80%).ConclusionsThe distribution of Class I integron in ESBLs-producingEscherichia coli is signiifcantly higher than that in non-ESBLs-producing strains, It is rational that Class I integron highly correlate with strong drug resistance in ESBLs-producing strains.

Objective To investigate the clinical outcomes of microwave heliotherapy in situ on the primary or metastatic malignant tumors of the pelvis.Methods From February 2000 to April 2009,18 patients with primary or metastatic malignant tumors of the pelvis were treated with microwave heliotherapy in situ,and followed a total or partial tumor resection.There were 11 males and 7 females with an average age of 45 years(range,16-72).Twelve cases were diagnosed as primary malignant tumor and 6 as metastases.Locations of tumors involved:the Ⅰ region 6 cases.the Ⅱ region 10 cases.and the Ⅲ region 2 cases.The exposures of all tumors were via aTtype or ilioinguinal approach.The lesions were heated at 50℃ for 20 min by 2450 MHz microwave,with surrounding soft tissue protected by copper.mesh.The necrotic tumor tissues were total or partial excised after treatment,with preservation of the anatomical continuity of the pelvic ring.Results The duration of surgery was 60-180 min (110 min on average).The blood loss was 400-800ml(480 ml on average).All patients were followed-up for 0.5-7 years(3.5 years on average).Tumor local recurred in 1 case with chondrosarcoma,and was survival in tumor-bearing after 6 months follow up.One case with malignant fibrous histiocytoma died due to brain,pulmonary,and all body metastases.One case with osteosarcoma died due to pulmonary metastases.Five cases with the metastases died due to non-pelvis metastases.Functions of hip joint in 18 patients were as follows:flexion 80°-130°,extension 0°-10°,abduction 25°-35°,and adduction 18°-23°.Conclusion The clinical result demonstrated that the advantages of microwave heliotherapy in situ were quick increase of temperature,sensitive responses,easy control of temperature,and effective inactivation of tumor cells in the malignant bone tumors of pelvis.

Objective To discuss the measurement of bone tumor volume on the basis of three dimensional images segmentation technology. Methods Twenty patients with lacunar bone tumor from Tianjin hospital and Tongji hospital were included in the study from January 2010 to August 2010. There were 11 males and 9 females. Each patient was exposed to spiral CT preoperatively. Then these primitive CT dates were imported into digital orthopedics clinical research platform (SuperImage orthopedics edition 1.1, Cybermed Ltd). The volume and maximum diameter of bone tumor were measured before operation by three-dimensional reconstruction technology. The actual tumor volume was measured during the operation. The tumor volume was also calculated from plain X-rays and CT scans as ellipsoidal or cylindrical depending on the tumor configuration and presence or absence of a soft tissue component. Results The tumor volume was measured to be (14.92±7.34) mm3, (16.65±6.95) mm3 and (34.29±15.70) mm3 using three-dimensional reconstruction technology, intraoperative elevation, and traditional radiograph measurement separately. It was found that there was no difference regarding the outcomes of measurement between three-dimensional reconstruction technology and gross intraoperative measurement. But obvious difference was detected between gross intraoperative measurement and traditional radiograph measurement. Coefficient of correlation between diameter and volume of bone tumor was 0.325 (P=0.162). Conclusion Digital measurement is a precise, efficient,convenient and repeatable method for bone tumor measurement.

Objective To explore surgical procedure of combined anterior-lateral and lateral approach for the treatment of bone tumors of femoral neck. Methods Forty patients with bone tumors of femoral neck treated in Tianjin Hospital were included from July 2005 to August 2009. Of the patients, 12 who were treated with curettage and bone graft through combined anterior-lateral and lateral incision were analyzed in this study. There were 7 males and 5 females with an average age of 34 years ranging from 17 to 68 years. 4 patients were diagnosed as chondroblastoma, 2 giant cell tumor, 3 fibrous dysplasia, and 3 single bone cysts. 7 patients suffered from pathologic fractures, and 5 had presented thin cortical bone because of tumor involvement. There were 1 tumor located in H1 zone, 4 in H2 zone and 7 in H1,2 zone according to ISOLS femoral neck classification. All patients were treated by curettage and bone graft via anterior-lateral approach, 10 cases underwent internal fixation with anatomical plate, and 2 cases with canulated screws with lateral approach. Results The follow-up time ranged from 10 to 68 months with an average of 35 months.Pain disappeared in all patients, and there were not recurrence of tumor, pathologic fractures and avascular necrosis. One case had complained of lateral femoral skin numbness which may be caused by injures of femoral lateral nerves. One case had difficulties in the valgus of hip joint. The mean MSTS score was 29.2 points ranging from 27 to 30 points. Conclusion Anterior approaches of "SP" incision is helpful to thorough curettage which decrease the risk of recurrence due to good visualization and intemal fixation is easy to perform via lateral approaches. The result suggested that combined anterior-lateral "SP" and lateral incision is liable option in treatment of bone tumors of femoral neck.

Objective To construct green fluorescent protein (GFP)-labeled pSELECT-GFP zeohBMP2 eukaryotic expression vector. Methods The encoding fragment of hBMP2 gene was obtained from a recombinant plasmid pcDNA3.1/CT-hBMP2 by using polymerase chain reaction (PCR). hBMP2 gene was inserted into pTA2-T-easy and pSELECT-GFPzeo-MCS eukaryotic expression vector, and then transferred into competence DHSα cells. After screening, pSELEC-GFPzeo-hBMP2 was obtained and identified by sequence analysis. The recombinant vector pSELECT-GFP zeo-rhBMP2 was transfected into CHO cells. The successful trasfection was verified by fluorescence microscope in 48-72 hours. The RT-PCR and immunofluorescence was used to confirm the hBMP2 expression. Western Blotting was used to detect the secretion of hBMP2.Results A 1216 bp fragment was obtained by PCR, the same as expectant fragment. The recombined pSE-LECT-GFPzeo-hBMP2 eukaryotic expression vector was identified by restriction mapping and sequence analysis. The results were identical with that of reported hBMP2 sequence (Genebank NM-001200). The successful transfection was verified by fluorescence microscope in 48-72 hours. The stable expression in eukaryotic cells was confirmed by immunofluorescence and RT-PCR which showed an obvious band between 1000-2000 bp. Western Blotting identified the immunogenicity of recombinant human BMP2 with the molecular weight of about 17×103. Conclusion The pSELECT-GFPzeo-hBMP2 eukaryotic expression vector was constructed successfully.

Objective To investigate the value of percutaneous transforaminal endoscopic spine system (TESSYS) in lumbar discectomy for disc herniation. Methods Thirty one patients with lumbar disc herniation were treated with TES-SYS and followed up 6-12 months. The involved levels of vertebral segment included L34 (2 cases), L45 (21 cases) and L5S1 (8 cases). The targeted puncture was performed under local anesthesia and fluoroscopic guidance. The foramen of involved level of vertebral segment was enlarged gradually with four trephinations, and the working cannula was inserted transforaminal in-to the canal. Then the herniation was exposed and removed with full endoscopic technique, including the loosen nucleus pulposus. The nerve root and dural sac were exposed and released adequately. Results The procedure was evenly carried out in 27 cases. After discectomy, the nerve roots were complete released, and not exposed in the first case of far lateral herni-ation and the second case of central herniation. The third case and eleventh case converted to microendoscopic discectomy, due to large herniation and intraoperative pain, respectively. The patients could walk in the same day, 1 or 2 days after opera-tion, with obvious relief of leg pain. One case of recurrence was found at 2 weeks after operation, who was treated conserva-tively. At the final follow-up, the visual analogue scale of leg pain decreased from 8.1±1.9 to 1.1±0.9, and the Oswestry dis-ability index (ODI) decreased from 31.1±8.3 to 3.4±3.3. According to MacNab scale, there were excellent results in 25 cases and good results in 6 cases. Conclusion The percutaneous endoscopic TESSYS is a good minimal invasive technique for lumbar discectomy, with good results and a learning curve.

BACKGROUND: Bone morphogenetic protein-2(BMP-2) production of targeted cells is promoted by transfection of adenoviral vectors containing gene, but there are some immune responses. Transfection with plasmid as vector holds promise.OBJECTIVE: To explore the feasibility to construct human bone morphogenetic protein-2 eukaryotic expression vector labeled with green fluorescent protein (GFP).DESIGN: Single sample observation.SETTING: Tianjin Hospital.MATERIALS: The experiment was performed at the Key Laboratory of Hormone and Development, Ministry of Health, Tianjin Medical University from March 2006 to March 2007. pcDNA3.1/CT-hBMP2 plasmid containing full-length hBMP2 gene fragment was provided by Dr. Li; bicistronic eukaryotic expression vector pSELECT-GFPzeo-MCS and Zeo was provided by Invivogen; pTA2(R)-T Easy by Dingguo, China; restriction enzymes BamHI and NheI, T4 DNA ligase by Jingmei Biotech; PCR upstream and downstream primer synthesis and sequencing by Augct, Beijing.METHODS: With pcDNA3.1/CT-hBMP2 as template, hBMP2 target fragment was subcloned by PCR binding with designed specific primers. The fragment was bound with pTA2-T-easy and pSELECT-GFPzeo-MCS, separately, and transfected into DH5 α cells. pSELECT-GFPzeo-hBMP2 containing GFP was obtained after screening.MAIN OUTCOME MEASURES: hBMP2 sequence was identified by PCR; whether hBMP2 was cloned into pTA2-hBMP2 and pSELECT-GFPzeo-MCS was identified by digestion and sequencing.RESULTS: A target fragment of 1 216 bp was obtained by PCR amplification, and cloned into pTA2-T-easy and pSELECT-GFPZeo-MCS. The screening and sequencing results showed that the target fragment was 100% matched with BMP2cDNA sequence (NM-001200) from GenBank.CONCLUSION: hBMP2 eukaryotic expression vector labeled with green fluorescent protein is successfully constructed.

Objective To analyze clinical features of patients with chikungunya fever and provide future reference for prevention and control of the disease. Methods Forty-six confirmed chikungunya fever inpatients were included. Their clinical symptoms, signs, blood count, key biochemical indicators and treatments were analyzed. The comparison between groups were done by ttest. Results The percentages of total cases presenting with fever, rash and joint pain were 100. 0% (46/46), 91. 3% (42/46) and 89. 1% (41/46), respectively. Fifteen (32.6%) cases displayed leucopenia. Increases in lactose dehydrogenase (LDH) and creatine kinase (CK) were observed in 45. 5%(20/44) and 28. 9%(13/45) of the cases, respectively. Three cases displayed an increase of alanine aminotransferase (ALT). Administration of ribavarin extend febrile time compared to symptom-relieving treatments (t=2. 588, P = 0. 013). Conclusions Clinical features of chikungunya fever include fever, rash and joint pain. Good prognosis can be resulted from symptom-relieving treatment. Antiviral treatment may not be beneficial to reducing course of disease.

Objective To investigate the methods and therapeutic effects of image-guided percutaneous needle iniection of methylprednisolone and injectable calcium sulfate for simple bone cysts.Methods Thirty-seven patients with simple bone cysts from 0ctober 2006 to August 2010 were analysed retrospectively in our hospital,including 26 males and 11 females with the average age of 13.2 years(range,8-22 years).Five cases of proximal femus lesions,with proximal thigh pain,limp and other symptoms.Thirty-two cases of proximal humeral lesions,16 patients had proximal pain and other symptoms of upper arm,the other 16 cases were asymptomatic.Preoperative AP and lateral X-ray.CT and MRl were taken.Under the C-arm X-ray monitor.two needles were inserted into the proximal and distal of cysts respectively,Omnipaque contrast was iniected to confirm the two needles is interlinked,then repeated rinsing with normal saline,then 120 mg methylprednisolone and iniectable calcium sulfate were injected,till the cysts were full up.Patients after treatment were assessed according to modified Neer X-ray criteria.Resuits The average hospitalization was 2.3 days (range.1-3 days).X-ray was reviewed every month,additional injection was performed if bone cysts stopped decreasing for 2 consecutive months,iniection 2 times in 6 eases,and 31 cases were injected only once.After 3 months follow.up 37 cases,according to modified Neer X-ray criteria,6 eases regarded as grade Ⅱ,8 as grade Ⅲ,23 as grade Ⅳ;after 6 months,31 patients were followed up,including 2 cases as grade Ⅱ,4 cases as grade Ⅲ,25 as grade Ⅳ;after 24 months of follow-up 26 cases,3 as grade Ⅲ,23 as grade Ⅳ;after 36 months follow-up,19 cases were all grade Ⅳ.Conclusion Imaging-guided percutaneous iniection of methylprednisolone and inieetable calcium sulfate for simple bone cysts has demonstrated,with less trauma,lower complications incidence and quicker recovery.

Objective To study clinical application of femoral head exclusion after resection of pelvic tumors around acetabulum with less limb damage and complications.Methods From October 1999 to August 2009,18 patients with pelvic tumors around acetabalum (zone Ⅱ of Ennekingl were treated with tumor resection and femoral head exclusion,including 16 males and 2 females with an average age of 54 years(range,18-72).The diagnosis were chondrosarcoma(4 cases),osteosarcoma(2),giant cell tumor of bone(4),synoviosareoma(1),malignant fibrous histiocytoma(1),Ewing sarcoma(1),and metastatic carcinoma(5).Throe cases were treated by selective arterial embolization before operation and 4 cases by temporary block of lower abdominal aorta or common iliac arte.Regular follow-up in clinic was done to observe function of joint and radiographic imaging.Results All patients were followed up for 55 months(range,3-118).The mean surgical time was 170 min(range,120-350 min).The mean blood loss was 1200 ml(range,600-2200 ml).All wounds were healed without infection.One patient with osteosarcoma and 3 with metastatic carcinoma died of multiple organ metastases during 6 months to 2 years after operation.The others had no local relapse.The mean limb discrepancy was 5.0 cm (range,2.0-7.5 cm).The mean Enneking score was 23 (range,20-27).All patients needed custom-made shoes with heightened heel of 2-3 cm.Osteonecrosis of the femoral head was not found in radiographic images.Eight cases had formed artificial acetabulum.One case had gentle lumbar scoliosis.Conclusion The clinical application of femoral head exclusion after resection of pelvic tumors around acetabulum had low complications,less pain,low relapse,better function in the recent period.