Objective To observe the efficacy of combinated inhaled glucocorticosteroid and long- ?2 - agonist for asthmatic children. Methods Forty- seven children with moderate or severe persistent asthma were treated with inhalation of the fluticasone pro-pionate and salmeterol. The PEF value and the mark according the symptom of asthma before and after treatment( week and month respectively) were recorded. At the same time the days of treatment the patient with the short - ?2 - agonist in the first week and the forth week respectively were noted. The subjective feeling and accident of patients also should be paid attention. Results There was significant difference (P

To clone the differential genes in antibiotic resistence Neisseria gonorrhoeae,the library that contains the fragments of differential genes between antibiotic resistance strain and standard reference strain of Neisseria gonorrhoeae was constructed which used suppression subtractive hybridization technique. Then the antibiotic resistance relational genes fragments were cloned and analyzed. Antibiotic resistance Neisseria gonorrhoeae subtractive library that has high subtractive efficiency was set up successfully. The amplified library contained 2500 positive clones. Sequence analysis was performed to find the fragments of antibiotic resistance relational genes. Five sequences were unknown previously. The fragments of antibiotic resistance genes may provide an important clue for studying the mechanism of occurrence and development of antibiotic resistance Neisseria gonorrhoeae.

ObjectiveTo analyze the characteristics of top of the basilar syndrome (TOBS) in clinic and imaging.MethodsData of 31 TOBS cases were analyzed retrospectively.ResultsThe clinical features of TOBS patients were sudden unconsciousness or vertigo and dyskinesia of the limbs, the dismovement of the eyeballs, abnormality of the pupils, partial blindness or cortical blindness, hypesthesia, disturbance of memory and counting. CT and MRI showed multi-infarction included thalami, occipital lobe, cerebellum, midbrain, temporal lobe.ConclusionTOBS can be diagnosed accurately according to clinical features and imaging signs.

OBJECTIVETo assess therapeutic effect of acupuncture at Sishencong (EX-HN 1) on insomnia.

METHODSSixty-five cases of insomnia were treated with acupuncture at Sishencong (EX-HN 1) and their quality and compositions of sleeping before and after treatment were evaluated with a multichannel sleep detector.

RESULTSAfter treatment, the quality of sleeping increased significantly (P < 0.05), and the sleeping rate increased from (61.95 +/- 22.90)% before treatment to (79.27 +/- 11.05)% after treatment (P < 0.05).

CONCLUSIONAcupuncture at Sishencong (EX-HN 1) has obvious therapeutic effect on insomnia.

This study was aimed to clone human intercellular adhesion molecule-1 (ICAM-1) gene, to transfect the constructed eukaryotic expression vector ICAM-1-GFP into CHO cells, as well as to detect ICAM-1-GFP expression in CHO cells binding with Molt-4 cells. ICAM-1 cDNA gene was amplified by RT-PCR and inserted in PMD(18)-T vector. Then ICAM-1 cDNA from pMD18-ICAM-1 vector was subcloned into eukaryotic expression vector pEGFP-C1 to construct recombinant ICAM-1-pEGFP-C1 vector. Restriction analysis and DNA sequencing were used to confirm the recombinant vector. After stable transfection of CHO-K1 cells with the recombinant vector, the expression and subcellular localization of ICAM-1-GFP were detected by RT-PCR, flow cytometry and fluorescence microscopy. The function of ICAM-1-GFP fusion protein was assessed by the binding of ICAM-1-GFP/CHO cells to Molt-4 cells. The results showed that 1622 bp full-length ICAM-1 cDNA obtained and was successfully ligated with pMD(18)-T-vector, subcloned to construct recombinant ICAM-1-pEGFP-C1 vector. Restriction analysis and DNA sequencing indicated that recombinant ICAM-1-GFP was successfully constructed and ICAM-1-GFP was expressed stably in CHO cells. ICAM-1-GFP expression was only observed in the cytoplasm of ICAM-1-GFP/CHO cells by fluorescence microscopy. The ICAM-1-GFP/CHO cells were bound to PMA-treated Molt-4 cells. The expression of MEM-148 was very weak in PMA-treated Molt-4 cells. It is concluded that the ICAM-1-GFP eukaryotic expression vector has been constructed successfully and expresses stably in CHO cells. PMA can increase the binding of Molt-4 cells to ICAM-1-GFP/CHO cells by inducing specialized form of ICAM-1 clustering.
Animals ; CHO Cells ; Cricetinae ; Cricetulus ; DNA, Complementary ; genetics ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; Recombinant Proteins ; genetics ; Transfection

The 1.7 kb full-length hemagglutinin (HA) gene fragment of H5N1 subtype avian influenza virus was amplified by RT-PCR and then cloned into the pFastBacHT donor plasmids. The recombinant plasmid pFastBac-H5 was identified by restriction enzyme digestion and sequencing. Following the transposition pFastBac-H5 into the bacmid in DH10Bac E.coli competent cells, the colonies were identified by blue and white selection. The recombinant bacmid (rBacmid-H5) was verified by PCR analysis. Transfection of rBacmid-H5 DNA into sf9 cells using Cellfectin reagent results in the production of recombinant viral stock. Cells were harvested 72h post infection and analyzed by SDS-PAGE, Western-blot, hemagglutination and hemagglutination inhibition test;the expressed HA protein (rH5)shows hemaggluting activity and can be inhibited by H5N1 virus immunized chicken sera. On the other hand, immunization of chickens with rH5 protein results in high titers of H5N1 virus specific hemagglutation inhibition antibodies, which proved its biological activity.

Objective To investigate the value of 99Tcm-methoxyisobutylisonitrile (MIBI) scintigraphy in assessing the preoperative chemotherapy response and multidrug resistance of osteosarcoma.Methods From January 2007 to October 2008, 12 patients (female:4, male:8; mean age:16.3 years,range:8-27 years) underwent early (10min) and delayed (120 min) 99Tcm-MIBI scintigraphy before and after preoperative chemotherapy.Seven cases had osteosarcoma at the distal femurs, 2 at the proximal tibias, 2 at the upper end of humerus and 1 at the fibula.The tumor-to-background ratio (T/B) and washout rate (WR) were calculated.Tumor necrosis was classified according to Huvos criterion after limb salvage surgery.Immunohistochemical staining for P-glycoprotein(gp) was examined.Spearman correlation analysis and t-test were performed.Results According to Huvos criterion, 7 patients were classified as good responders with more than 90% of tumor cell necrosis and 5 as poor responders with less than 90% of tumor cell necrosis.R value (ratio of early phase T/B after and before chemotherapy) was significantly lower in good responders than that in poor responders (0.473 ± 0.21 vs 0.998 ± 0.06, t= 5.342, P= 0.000 ).R value was significantly correlated with the degree of tumor cell necrosis ( rs=- 0.87, P= 0.000 ).WR was significantly higher in patients with positive P-gp expression than that in patients with negative P-gp expression ((38.36 ±18.64)% vs (6.40±5.87)%, t= -3.278, P=0.008).There was significant correlation between the WR and P-gp expression (rs = 0.91, P= 0.001 ).Conclusion 99Tcm-MIBI scintigraphy is a feasible non-invasive technique to assess the chemotherapy response and to detect P-gp expression of osteosarcoma.

Objective To analysis and determine the possibility of the Citellus undulatus infected with Yersinia pestis surviving the winter in an experimental study, and to provide scientific experimental basis for the study on the mechanism of Yersinia pestis preservation. Method In 2006,09 to 2007,04 and 2007,09 to 2008,04 in Xinjiang Wusu-Gurtu natural foci of plague, under natural conditions, the over the winter process of Citellus undulatus carrying the plague bacteria was simulated, and 178 Citellus undulatus were infected with Yersinia pestis (1×107 Bacteria/mouse) using artificial injection method. One hundred seventy-eight Citellus undulatus infected with Yersinia pestis were kept into a construction of the black (1-5 ℃) basement (2 meters under the ground) in the plague focus. In doing so, these Citellus undulatuses almost simultaneously stepped into hibernation. After waking up from hibernation in following year in April, the survived mice carrying the plague bacteria were observed. Results Sixty-eight mice survived among the 178 infected with Yersinia pestis after 6 months of hibernation (through October to the following year in April), and the remaining 110 were all dead without pulling through the hibernation period. The survival rate was 38.2% (68/178). The organ culture of Yersinia pestis of the 110 dead mice(Citellus undnlatus) were tested, 67 were negative(-), 43 positive(+), with a positive rate of 39.1%(43/110). Among the rats with positive plague bacteria, the congestive pulmonary edema and the pathological changes of the hemorrhagic inflammation of the heart, liver, spleen, kidney and injection site could be seen clearly; the plague-free mice were not found to have any pathological changes. The survived 68 mice over the winter were autopsied and observed after being fed up for 20 days. No any pathological changes were found among these mice, and culturing of Yersinia pestis of the heart, liver, spleen, lungs and the tissue of injection site of these mice were all negative (-). Conclusions Citellus undulatus can carry Yersinia pestis during hibernation, but some fail to carry the bacteria through the entire process of hibernation persistently. Yersinia pestis was negative in the survived mice at the end of hibernation. The results showed that Citellus undulatus can not carry Yersinia pestis over the winter.

Aim The effect of tetrandrine on TGF-?1 mRNA expression in glomerulosclerosis rat was observed. Methods The rats were randomly divided into four groups, such as the normal control group (sham operative rat), glomerulosclerosis model group,tetrandrine group and amlodipine group. The expression of TGF-?1 mRNA was analyzed by Northern blot hybridization. Results The expressions of TGF-?1 mRNA in two treating groups were much lower than untreated model group. There were no difference between these two treating groups. Conclusion Tetrandrine can decrease the expression of TGF-?1 mRNA in glomerulosclerosis rat induced by unilateral renctomy plus adriamycin.

AIM: To investigate the expression of cystic fibrosis transmembrane conductance regulator (CFTR) in acute myeloid leukemia (AML) and its effect on the biological function of human erythroleukemia cell line TF1, and to explore the underlying mechanism .METHODS: The abundance of CFTR in the bone marrow mononuclear cells of patients with AML was measured by real-time PCR.After TF1 cells were incubated with CFTR specific inhibitor CFTRinh-172, cell viability, cell cycle distribution and cell apoptosis were analyzed by CCK-8 assay and flow cytometry . The Wnt signaling pathway-related proteins were determined by Western blot .RESULTS: CFTR was highly expressed in both patients with AML and leukemia cell lines .After incubated with CFTRinh172, the viability of TF1 cells was de-creased, the proportion of the cells in G0/G1 phase was increased, while that in S phase declined (P<0.05).Further-more, the cells treated with CFTRinh-172 exhibited higher apoptotic rate , accompanied with lower protein expression of β-catenin, c-Myc and cyclin D1 (P<0.05).CONCLUSION:CFTR expression is dramatically increased in AML .Inhibi-tion of CFTR restrains the growth and promotes the apoptosis of TF 1 cells via classical Wnt signaling pathway .