Objective To constitute a higher expression system and to obtain the breast cancer cell strains in which BSP is stably expressed. Methods hBSP gene was subcloned from pB-hBSP vector by PCR. The PCR fragment was inserted into the eukaryon expression vector, pIRES2-EGFP, which allow exogenous protein and EGFP to express respectively. The recombinant vector, pIRES2-hBSP-EGFP, was transfected into human breast cancer cells with Lipofectamine TM 2000. The expression of symbol protein EGFP, could be conveniently observed with fluorescent microscope. Results The recombinant pIRES2-hBSP-EGFP plasmid was constituted and successfully transfected into breast cancer cells. In the breast cancer cell strain hBSP and EGFP were expressed. Conclusion The successful constitution and transfection of hBSP and EGFP nonfusion expression vector laid a foundation for the further study on the effect of BSP on breast cancer metastasizing to bone in vivo or in vitro.

Objective To explore the effect of thymosin and MVP chemotherapy on the life quality of non-small cell lung cancer(NSCLC) patients. Methods 50 cases of NSCLC patients who received MVP(MMC, VDS, DDP) chemotherapy,were randomly divided into experimental group (using thymosin α1) and control group;before chemotherapy and at the end of chemotherapy (the first 2 weeks), the quality of life was evaluated and analyzed comparatively by the FACT-L questionnaire of patients with lung cancer-Chinese version (V4.0). Results The re-sults of the experimental group score increased by (3.13±2.29),and control group score increased by(-1.07± 2.19) with significant differences (P<0.01). Conclusions Thymosin α1 can improve the short-term quality of life of the non-small cell lung cancer patients who received MVP chemotherapy.

Acute Disease ; Adolescent ; Adult ; Aged ; Cyclophosphamide ; administration & dosage ; therapeutic use ; Female ; Hemoperfusion ; Humans ; Male ; Methylprednisolone ; administration & dosage ; therapeutic use ; Middle Aged ; Paraquat ; poisoning ; Renal Dialysis ; Retrospective Studies ; Young Adult

BACKGROUND: Bone sialoprotein (BSP) gene is expressed in human breast cancer cells, in which bone metastasis occurs easily outside the mineralized tissue. Clinical observation shows that the expression level of BSP of breast cancer cells at bone metastasis is higher that at the primary site;therefore, BSP may be closely related to tumor specific bone metastasis.The study on breast cancer bone metastasis can provide new drug target for clinical prevention and treatment.OBJECTIVE: To establish breast cancer cell strains of BSP with stable expression and observe the effect of BSP in the whole process of breast cancer bone metastasis.DESIGN: Controlled experiment.SETTING: College of Biological Sciences and Engineering, South China University of Science and Technology; Medical Experiment Center,Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA.MATERIALS: This experiment was conducted in the Medical Experimental Center,Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA,betweer November 2003 and March 2004..pIRES2-EGFP vector (5.3 kb) was purchased from BD Biosciences Clontech Inc.; E.Coli.Top10, pB-hBSP plasmid containing the coding region of hbsp, and human breast carcinoma cells, MDA-MB-231BR that was specifically transferred to brain and MDA -MB-231BO that was specifically transferred to bone.METHODS: hbsp gene was subcloned from pB-hBSP vector by PCR. Bg1Ⅱ and Pst Ⅰ restriction enzyme sites were inserted at 5' and 3' ends, orientation cloned to eukaryon expression vector pIRES2-EGFP, and constructed recombinant vector pIRES2-EGFP. The constructed recombinant vector was transfected into MDA-MB-231BR that was specifically transferred to brain and MDA-MB-231BO that was specifically transferred to bone.MAIN OUTCOME MEASURES: Construction of pIRES2-hBSP-EGFP recombinant expression vector; recombinant expression vector pIRES2-hBSP-EGFP transfecting breast cancer cells.Breast cancer strains specific in bone metastasis and brain metastasis were successfully transfected. The fluorescence labeling could be observed under the fluorescence microscope, and BSP had corresponding expression.CONCLUSION: The successful construction and transfection of pIRES2hBSP-EGFP of eukaryon expression vector would lay foundation for further study on the role of BSP in breast cancer metastasizing to bone in vivo or in vitro.

The effect of reaction media cosolvent water activity, temperature and pH on Novozym 435-caulyzed enantioselective esterification of ketoprofen was systematically explored. Novozym 435 showed high catalytic activity and enantioselectivity in cyclohexane; E value increases markedly by addition of toluene to cyclohexane; the optimum temperature and the initial water activity were found to be 30℃ and 0.09 respectively; pH shows little effect on enzymatic reaction wilson the scope studied.

OBJECTIVETo analyze the causes of delayed union or nonunion of the ulna after intramedullary nailing in pediatric forearm fractures.

METHODSFrom February 2005 to February 2010,5 patients with forearm fractures who were treated with titanium elastic nailing (TEN) were identified to fulfill the criteria of having developed a delayed union or nonunion of the ulna. The causes of delayed union or nonunion were investigated according to mechanism of injury, fracture location, treatments methods and postoperative management. All patients were male and the age was 3 to 14 years old with an average of 9.4 years. All fractures were located on the mid-third part of forearm. Two cases had a re-fracture. Among them, 3 cases caused by high-energy injury and 2 cases by falling down. Open reduction were performed in 4 cases while the other one was treated with closed reduction. Four patients were immobilized in an above-elbow cast, postoperatively.

RESULTSAll patients were followed up from 7 to 19 months with an average of 11.4 months. There were 4 delayed union and 1 nonunion. Three patients healed after the removal of the nail and avoidance of weight-bearing. Two patients healed by replacing another fixation. No patients had soft-tissue irritation or nail-entry-site infections.. The clinical effect was evaluated according to Daruwalla and Price scores with 3 excellent and 2 good of the results.

CONCLUSIONSUsing titanium elastic nailing for the treatment of pediatric both-bone forearm fractures is a good method. However,strict indication selection should be followed to avoid delayed union or nonunion.

Adolescent ; Child ; Child, Preschool ; Fracture Fixation, Intramedullary ; methods ; Fracture Healing ; Humans ; Male ; Radius Fractures ; physiopathology ; surgery ; Retrospective Studies ; Ulna Fractures ; physiopathology ; surgery

BACKGROUND: The cells of biomaterial compound cytokine gene or compound transfected cytokine gene that are transplanted into the area of bone defect can promote bone repair.OBJECTIVE: To investigate the feasibility of gene therapy for bone defect after rat transforming growth factor (TGF)β1 gene is transfected into osteoblasts.DESIGN: A controlled and observational experiment.SETTING: Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: Five newborn Sprague-Dawley rats of either gender were included.METHODS: The experiment was conducted in the laboratory of Orthopedic Department, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, from February to September 2000.Rat osteoblasts were transfected with pcDNA3-TGF-β1 plasmid by lipofectamine mediated gene transfer, and the plasmid pcDNA transfected cells were set as control group. The transient expression of TGF-β1 was detected by strept avidin-biotin-peroxidase complex (SABC) method and in situ hybridization detection 24 hours later. The cells transfected by G418 for 2 weeks were detected with SABC to investigate the stable expression of TGF-β1 gene.MAIN OUTCOME MEASURES: SABC method and in situ hybridization detection were applied to detect gene expression.of pcDNA3-TGF-β1 transfected osteoblasts and in situ hybridization detection: After the osteoblasts were transfected for 24 hours, cytoplast was full of brown granules and there were no brown granules in the cytoplast of blank carrier transfected cells, indicating that TGF-β1 mRNA was signifiG418: transfected cells still had high expression of TGF-β1 after 2-week G418 screening.CONCLUSION: Osteoblasts can express cytokine immediately with high effect using gene transfection technique. The stable and high expression is presented after TGF-β1 gene is transfected into osteoblasts at the moment of transfection and after 2-week screening, proving the feasibility of gene therapy for bone defect when cytokine gene is transfected into osteoblasts.

Objective:To evaluate the immunogenicity of an experimental SARS-CoV inactivated vaccine.Methods:The virus suspension of F69 strain was inactivated with 0.4% formaldehyde and purified,then used as the immune antigen combined with Freund′s adjuvant.Eight adult New Zealand rabbits were immunized 4 times with this vaccine.12 sets of rabbit serum were sampled from the third day to 74th day after first immunization.Titers of specific IgG antibody and neutralizing antibody were determined by indirect ELISA and micro-cytopathic effect neutralizing test.Results:Rapid and potent humoral immune responses were induced by F69 inactivated vaccine in all eight immunized rabbits.Both specific IgG antibody and neutralizing antibody all peaked just with 2 vaccinations,with the maximum titer of 1∶81 920 and 1∶20 480 respectively about 6 weeks after first immunization.Across neutralizing reaction existed between F69 and Z2-Y3 strains.Conclusion:F69 inactivated vaccine owns strong immunogenicity.Similar antigenic epitopes exist between the F69 strain and Z2-Y3 strain,which ensured the cross neutralizing reaction.

Objective:To explore the rules of prescription and medication of Traditional Chinese Medicine (TCM) syndrome differentiation treatment for insomnia.Methods:We screened the medical records related to insomnia from the ancient medical cases, modern medical cases, famous medical cases and shared medical record database, and then we put data into the Ancient and Modern Medical Case Cloud Platform (V 1.5.7). The platform software Data mining was applied and performed by frequency analysis, correlation analysis, and drug pair analysis. Results:This study collected 664 cases related to insomnia, like difficulty in falling sleep, early waking up, easy to wake up, dreaminess. The top 5 syndromes included internal phlegm-heat, imbalance between heart-yang and kidney-yin, liver stagnation transforming into fire, heart and spleen deficiency, restless mind. There were 664 prescriptions, 414 TCM medications, where the top 5 medications included Glycyrrhizae Radix et Rhizoma, Poria, Rehmanniae Radix, Ziziphi Spinosae Semen, Angelicae Sinensis Radix. The effect of high-frequency medications were nourishing the heart and calming the mind, clearing heat and dissipating phlegm, soothing the liver and relieving depression, and nourishing yin and yang. The top five drug-combinations included Ostreae Concha- Longgu, Poria- Glycyrrhizae Radix et Rhizoma, Citri Reticulatae Pericarpium- Poria, Angelicae Sinensis Radix- Rehmanniae Radix, Bupleuri Radix- Glycyrrhizae Radix et Rhizoma. Conclusions:TCM syndrome differentiation for insomnia focus on the heart. The medications are mainly to nourish the heart and calm the mind, supplemented by clearing heat, replenishing heart and spleen, soothing the liver and relieving stagnation, nourishing yin and blood for the symptoms relief, with treatment priciple of seeking both temporary and permanent solutions.

Objective The aim of this study was to evaluate the negative predictive value (NPV) and Sensitivity(Sen) of ruling out acute myocardial infarction(AMI) using low risk electrocardiogram(ECG) and baseline high-Sensitivity troponin T(hs-cTnT) immediately in chest pain patients. Methods Patients presenting to chest pain center with complain of chest pain in West China hospital of Sichuan university were enrolled. Clinical data including ECG and hs-cTnT concentration were gained .According to different diagnosis cutoff point, the NPV and Sen of ruling out AMI using low risk ECG and baseline hs-cTnT immediately in chest pain patients were evaluated. Results An hs-cTnT cutoff of 5 ng/L resulted in a NPV of 99.9%ruling out of all non-AMI patients. Adding the information of a low risk ECG resulted in a 100%NPV (19.51%ruled out). At any diagnosis cutoff point, present hs-cTnT concentration can not rule out AMI completely . Adding the information of a low risk ECG , the NPV of present hs-cTnT was improved and more non-AMI patients were ruled out safely. Conclusions It is safe to rule out AMI among the chest pain patients when the level of hs-cTnT less than 5 ng/L combined with low risk electrocardiogram.