Objective To develop a method to determine lead in the whole blood with graphite furnace atomic absorption spectrometry.Methods The whole blood was diluted with mixing matix modifiers [5.0 g/L NH4H2PO4+1.0 g/L Tritonx-100+2.0 ml/L HNO3],then the samples were automatically injected and the blood lead level was determined with Zeeman graphite furnace atomic absorption spectrometer.Results There was significant linear relation between absorbance and lead level in the range of 0-40 ?g/L(r=0.998 2),the relative standard deviation(RSD)was 5.5%-12.2%,the recovery rate was 90%-106%,detection limit was 1.1 ?g/L.Conclusion This method is characterized by reliable result,convenient procedure as well as lower pollution.The test result of whole blood lead level in more than 600 occupational lead exposed people with this method is satisfactory.

Objective: To study the interpersonal interaction of junior high school students and its relationship with self-consciousness. Methods: 265 junior high school students were given the measurement of interpersonal relationship, and administered the questionnaire of Piers-Harris children self-consciousness one week later. Results: ①The interpersonal relationship of junior high school students was interactive. ②There were statistically significant differences among some dimensions of self-consciousness of junior high school students of different grades and genders. ③Intellgence and school performance had important effect on personal acceptance of junior high school students. Conclusion: There is interpersonal interaction among junior high school students, and intelligence and school performance influence the personal acceptance significantly.

ObjectiveTo investigate the effects of epidermal growth factor (EGF),fibroblast growth factor(FGF), and bovine serum on proliferation and apoptosis of the cultured fetal human retinal cells.MethodsEGF and FGF were added or not to the medium of fetal human retinal cells cultured by bovine serum in vitro. The number of cells, bromodeoxyuridine(BrdU) incorporation and Tdt-mediated dUTP nick end labelling(TUNEL) were detected to determine the proliferation and apoptosis. Immunohistochemical staining of neuron specific enolase(NSE), Thy1.1, glial fibrillary acidic protein(GFAP) and scan electromicroscopy were performed to identify cell components. The expression of transcription factor c-fos, c-jun and apoptosis regulation factor bcl-2 and Bax were examined by immunohistochemical staining to explore the underlying mechanism.ResultsThe increased number of NSE and Thy1.1 positive cells and BrdU incorporation, and decreased apoptotic cells were found in the groups treated with EGF and FGF. Meanwhile, the up-regulation of c-fos, c-jun and bcl-2 were also found.ConclusionEGF and FGF can promote the survival and proliferation of cultured retinal cells by up-regulating the expression of c-fos, c-jun and bcl-2.

Objective To compare the effects of continuous versus intermittent feedings on the growth of very low birth weight infants. Methods Databases of Pubmed,Embase,Ovid,the Cochrane library and CBM were searched through internet,and citations of relevant original studies were also searched manually by using keywords of 'ontinuous,intermittent,nasogastric,premature,very low birth weight' Meta analysis was done on the results of these studies. Results Seven eligible studies with 434 infants were identified,among them,217 were fed using continuous nasogastric gavage,and other 217 were fed by intermittent nasogastric gavage. The result of the Meta analysis revealed that there was no statistical difference in the growth of the infants between two feeding methods. Conclusions Feeding methods are associated with similar outcomes when calorie intake is guaranteed.

50% response) in 3 cases(10%),poor in 3 cases. Conclusions Ultrasound-guided intralesion injection of Bleomycin is effective therapy for cystic lymphangioma in children.

Objective To investigate the transfection efficiency of combination of ultrasound (US) microbubbles (MBs) mediated liposome small interference RNA (siRNA) to human and rat retinal pigment epithelium (RPE) cells. Methods Human and rat RPE cells and siRNA were incubated in 24-well plates (2×10~5/well and 1×10~5/well, respectively). The cells were devided into 5 groups: siRNA+US, siRNA+MBs+US, siRNA+L (liposome), siRNA+L+US, siRNA+L+US+MBs. After 12 h, transfection efficiency was observed with fluorescence microscopy and flow cytometry. Results US or ultrasound targeted microbubbles destruction without liposome-mediated could not promote siRNA transfection efficiency to human and rat RPE cells. The transfection efficiency of human and rat RPE cells significantly decreased in the siRNA+L+US+MBs group, but increased in siRNA+L+US group. Conclusion Ultrasonic irradiation can promote lipid-mediated siRNA transfected human RPE cells.

BACKGROUND: Genetically engineered cells have been used in the replacement therapy and gene therapy. However, how to select proper donor cells, target cells, and corresponding viral vectors is the most difficult in the therapy.OBJECTIVE: To compare the transduction efficiency of recombinant adenovirus Ad5 and AdSF35, adeno-associated virus rAAV1/2 and rAAV2, and lentivirus LV in bone marrow mesenchymal stem cells (BMSCs) and exogenous gene expression level so as to select the vectors, which can efficiently transduce BMSCs.DESIGN, TIME AND SETTING: Gene engineering controlled observation, performed in the Central Laboratory, Shanghai First People's Hospital between October 2006 and March 2007.MATERIALS: Ten Sprague Dawley rats of clean grade were used to prepare BMSCs. All recombinant viral vectors carded enhanced green fluorescent protein (EGFP) report gene. Ad5 was prepared by the Central Laboratory, Shanghai First People's Hospital. Ad5F35 was gifted by professor Qian Qi-jun from the Second Military Medical University of Chinese PLA. rAAV2 and rAAVI/2 were the products of Benyuan Zhengyang Gene Technique Co.,Ltd. LV was gifted by professor Cuo Li-be from Shanghai Institute of Biochemistry and Cytobiology, Chinese Academy of Sciences.METHODS: Rat BMSCs were in vitro isolated and cultured by density gradient centrifugation. BMSCs of passage 4 were inoculated into 24-well plate at lxlO5/well. One day later, ceils adhered to the wall and allocated to 5 groups. Ad5-EGFP [10, 100,1 000 multiplicity of infection (MOI)], Ad5F35-EGFP (10,10, 1 000 MOI), rAAVI/2-EGFP (1×104,1x10× vg), rAAV2-EGFP(1×104, 1x105vg), and LV-EGFP (30 TU) were respectively added into the 5 groups. BMSCs were transduced for 2 days with Ad virus and separately for 6 days with rAAV and LV virus.MAIN OUTCOME MEASURES: EGFP-positive expression and fluorescence intensity.RESULTS: After twenty-four hours of Ad5-EGFP transduction, EGFP-positive cells were visible under the microscope. With virus dose increasing, EGFP-positive cells increased and fluorescence intensity strengthened. Twelve days later, EGFP-positive cells gradually reduced and fluorescence intensity weakened. For Ad5F35-EGFP, its transduction was basically similar to Ad5-EGFP, but EGFP-positive cell number and fluorescence intensity were increased. After 6 days of rAAV1/2-EGFP or rAAV2-EGFP transduction, EGFP-posidve cell number and fluorescence intensity were decreased. For LV-EGFP transduction, a small amount of EGFP-positive cells could be visible on the second day, and then EGFP-positive ceils and fluorescence intensity were gradually increased until the sixth day.CONCLUSION: Ad5, Ad5F35 and LV could effectively transduce BMSCs cultured in vitro and express exogenous gene.Furthermore, transduction efficiency was correlated with virus dose in dose-dependent manner, rAAVI/2 and rAAV2 had poor/in vitro transduction efficiency.

Objective: To investigate the effect of cobalt chloride (CoCl2) on transgene expression and viral particle titers in tumor cells infected by conditionally replicating adenovirus expression vector with hypoxia response elements(HRE)-regulated E1AE1B expression (Ad-5HRE-E1AE1B-RFP) and non-HRE regulated replication-deficient adenovirus expression vector (Ad-EGFP, Ad-Luc) in vitro and in vivo. Methods: Ad-5HRE-E1AE1B-RFP had five duplicated HRE and mini CMV acted as a promoter to drive E1AE1B expression and constitutive expression of RFP as reporter. The hypoxia model was optimized by exposing tumor cells to different concentrations of CoCl2. The hypoxia-inducible factor 1 alpha (HIF-1α) protein expression was determined by Western blotting. Under the optimized hypoxia model, the positive expression of exogenous gene and virus replication of Ad-5HRE-E1AE1B-RFP or Ad-EGFP-infected tumor cells were examined by conversed microscopic observation, FACS analysis and plaques formation test. Furthermore, transgene expression induced by combined application of hypoxia-inducible replicative adenovirus and replication deficient adenovirus (Ad-Luc) was also evaluated by examining the lucifererse activity in xenografted tumor models in nude mice by micro PET. Results: Western blotting results showed that CoCl2 at 0.4 and 0.08 μg/mL could stabilize and acumulate HIF-1α protein in gastric cancer SGC7901 cells, which could better mimic hypoxia condition. The microscopic observation and FACS analysis showed that CoCl2 at 0.4 μg/mL could remarkably increase the transduction efficacy of Ad-5HRE-E1AE1B-RFP, which was verified by significant increase in the percentage of positive expression of exogenous gene RFP and fluorescence intensity. But plaques formation test showed that Ad-5HRE-E1AE1B-RFP had no replication. CoCl2 0.4 μg/mL augmented the tranduction efficacy and expression levels of non-HRE regulated replication deficient adenovirus Ad-EGFP and Ad-Luc. Combined intratumoral injection of Ad-5HRE-E1AE1B-RFP and Ad-Luc significantly increased the expression of Ad-Luc in nude mice.Conclusion: CoCl2 markedly enhances transgene expression of recombinant adenovirus. However, the underlying mechanism is not only related to the CoCl2-induced hypoxia, but also probably related to regulation of gene transcription.

Objective To investigate the relationship between QRS wave terminal distortion with coronary arterial lesion and serum high-sensitivity cardiac troponin I (hs-cTnI) in early stage of acute ST-segment elevation myocardial infarction (STEMI).Methods One hundred and twenty patients with STEMI were classified into the QRS wave distortion positive group(QRS+,n=81) and non-QRS wave distortion group(QRS-group,n=39) according to EKG on admission.The two groups all conducted the coronary angiography and hs-cTnI detection.The coronary arterial lesion occurrence situation and hs-cTnI level were compared between the two groups.Results (1) In the QRS+ group:68 cases (83.59％) were male and 13 cases (16.05％) were females;in the QRS-group:27 cases(69.23％) were male and 12 cases (30.77％) were female.The sex difference had statistical significance (P＜0.05).(2) The occurrence rate of left anterior descending artery (LAD) lesion in the QRS+ group was higher than that in the QRS-group,the difference was statistically significant (P＜0.05).But the occurrence rate of left circumflex coronary artery (LCX) lesion in the QRS-group was higher than that in the QRS+ group,the difference was statistically significant (P＜0.01).(3) The hs-cTnI level in the QRS+ group was higher than that in the QRS-group,the difference was statistically significant (P ＜0.01).Conclusion The patients with QRS wave distortion positive have a higher occurrence rate of LAD lesion,while the patients with out QRS wave distortion negative have higher occurrence rate of LCX lesion;the QRS wave terminal distortion has relationship with serum hs-cTnI level.

A fusion gene called Ig-BDNF, in which brain-derived neurotrophic factor cDNA fused to the 3' end of signal peptide of Ig coding sequence, was constructed by PCR, digested and subcloned into shuttle plasmid pSNAV to obtain a recombinant plasmid pSNAV-Ig-BDNF. Then the plasmid encoding fusion protein was transfected into 293 cell lines and the stably transfected clones were selected with neomycin. AAV1 containing Ig-BDNF fusion gene vectors were obtained by super-infection by Herpes virus. The resultant adeno-associated virus vectors AAV-Ig-BDNF were confirmed by PCR, Western blotting and a sandwich enzyme-linked immunosorbent assay (ELISA) after infection of 293 cell lines. The results indicated that AAV-Ig-BDNF contained the target gene, and infected cells and produced the fusion protein into the supernatant. The content of BDNF in medium per 5x104 cells over a 24 h incubation period reached 1000 pg/mL. With the help of non-replicative adenovirus during AAV-Ig-BDNF infection, the expression of BDNF increased 7-8 fold, and the enhancement of BDNF gene expression was observed in a concentration-dependent manner. These results suggested that a functional AAV-Ig-BDNF was successfully constructed and it offers basis for further study for gene therapy of neural degeneration diseases.
Brain-Derived Neurotrophic Factor ; biosynthesis ; genetics ; Cell Line ; Dependovirus ; genetics ; metabolism ; Genetic Vectors ; genetics ; Humans ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Recombination, Genetic ; Simplexvirus ; genetics ; Transfection