Objective To discover the main factors influencing the hospitalization expenses incurred by cases of community acquired pneumonia(CAP). Methods The hospitalization expenses incurred by 362 cases of CAP treated by the Department of Respiratory Medicine of the authors hospital from 1999 to 2000 as well as the composition of the expenses, the expenses for testing pathogens and the use of antibiotics were analyzed retrospectively. And the influencing factors of the hospitalization expenses were studied by means of stepwise regression. Results The average CAP hospitalization expenses were 9 253 yuan, with the expenses for medicine accounting for 51.4%. Among the antibiotics used, ? lactam was most frequently used. Next came quinolone and macrolides. The expenses for testing CAP pathogens were high while the positive rate was low. The major factors influencing CAP hospitalization expenses were respectively length of stay, time of intravenous drip of antibiotics during hospitalization, incidence or no incidence of heavy pneumonia, and the number of basal disease entities(P

ObjectiveTo study the cloning and sequencing of mature fragment of human bone morphogenetic protein-4 gene. Methods The template DNA was obtained from the human osteosarcoma cell line U2OS. By using RT- PCR method, the cDNA coding for the mature fragment of BMP-4 was amplified, cloned into the vector pUC19, and sequenced by Sanger Dideoxy-mediated Chain Termination method. Results The mature fragment of BMP4 cDNA was obtained by RT-PCR and determined by sequencing. Through the computer search on Genebank, the analysis showed that the homology of nucleotides and amino acids between cDNA of rhBMP4 mature fragment of this study and the published sequence was 99％. Sequence analysis showed that there were two differences, one was at base 1154 (201): G→C, which had no influence on the corresponding amino acids (Val). Another was at basel222 (269):C→T, the mutation at the base 1222 had the change of Ala to Val. Conclusion The mature fragment of BMP4 gene has been cloned. The results will be of great significance in treatment of skeletal injuries and diseases.

OBJECTIVE: To observe the effects of ginsenoside (Gs) and berberine (Ber), two kinds of active components of traditional Chinese herbal medicine, on transforming growth factor-beta1 (TGF-beta1) and prostaglandin E2 (PGE2) in PG cells. METHODS: Co-culture system of human lung carcinoma cell line PG and human T lymphocyte cell line Jurkat was established. PG cells were treated with Gs (100 microg/ml) and Ber (10 mug/ml) for twenty-four hours, and then cocultured with Jurkat cells. After 24-hour coculture, the state of Jurkat cells was observed with inverted microscope. The viable count of Jurkat cells was detected by trypan blue staining after 6- and 24-hour coculture, and the apoptosis of Jurkat cells was evaluated by flow cytometry. PG cells were treated with 100, 50, 25 microg/ml Gs and 10, 5, 2.5 microg/ml Ber respectively, and the content of TGF-beta1 and PGE(2) in PG cells was detected by enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA) method. RESULTS: After coculture with PG cells treated with Gs and Ber, the number of Jurkat cells was less than blank control group, and the apoptosis rates of Jurkat cells in Gs- and Ber-treated groups were higher than blank control group. Gs and Ber could promote the secretion of TGF-beta1 in PG cells, but could not change the level of PGE(2). CONCLUSION: Gs and Ber can promote the growth inhibition and apoptosis of Jurkat cells induced by PG cells, which may be related to the up-regulation of Gs and Ber on TGF-beta1 secretion in PG cells.

Objective To establish an LC-MS/MS method for the detection of landiolol concentration in human blood.Methods After pretreatment with neostigmine and a deproteinization procedure, landiolol and the internal standard venlafaxine were eluted isocratically using a mobile phase consisting of acetonitrile and 10 mmoL·L-1 ammonium acetate with 0. 1% formic acid in a ratio of 3664 ( V/V ) . Separation of the respective compounds was achieved on a Waters XTerra? RP18 column (150 mmí4. 6 mm,5 μm). Quantitative analysis of landiolol was conducted by a triple-quadrupole mass spectrometer with positive-electrospray ionization source,monitored under a multiple reaction monitoring ( MRM) mode. The extracted ions monitored following MRM transitions were m/z 510. 5→423. 1 for landiolol and m/z 278. 2→215. 1 for the internal standard venlafaxine. ResultsThe calibration curve of landiolol in human blood showed good linear relationship in the range of 1. 010-2 020 μg·L-1 . The lower limit of quantitation was 1. 010 μg · L-1 . The RSD of within-day and between-day precision was less than 6. 5% and 4. 8%, respectively. The recovery rate was 92. 6%-100. 9%. Conclusion The method is proven to be simple,rapid and reliable,and can be applied to study the pharmacokinetics of landiolol hydrochloride in healthy Chinese volunteers.

AIM: To investigate the effect of phellodendron and three kinds of its main components, which have asuppressive effect on the immune system, on the membrane fluidity of normal murine splenocytes. METHODS: The fluidity ofmembrane lipid regions of splenocytes was determined by the fluorescence polarization technique using 1, 6 - diphenyl - 1, 3, 5- heatriene (DPH) as a fluorescence probe. RESULTS: The results showed that the water extract of phellodendron and one of itsmain components (palmatine) increased the cell membrane fluidity in the inactive state, but the other two components, berberineand jatrorrhizine, decreased the cell membrane fluidity. After activated by ConA, all of them can decrease the cell membrane flu-idity. CONCLUSION: These results suggest that their immunosuppressive function might be due to decreasing the cell membranefluidity.

AIM: To observe the effects of some component of Chinese herbs for external use on proliferation of human umbilical vein endothelial cells (HUVEC) and investigate the mechanism of promoting tissue repair. METHODS: The method of MTT was used to examine the effects of Rg1, Rh1, perlolyrine, cinnamyl aldehyde, muscone, astragalus polysaccharin (APS), velver antler polypeptide (VAP) and soluble extract of boswellia carterii birdw (BCB) on proliferation of HUVEC. RESULTS: APS did not promote proliferation of HUVEC at 9.75 mg/L-2.5 g/L; Rh1 promoted proliferation of HUVEC at 1.94 mg/L-0.5 g/L (P

Objective To investigate the treatment of spinal cord injury (SCI) by recombinant adeno-associated virus (AAV) transducing the hNGF gene, and construct and produce the vector of hNGF recombinant AAV. Methods The resulting gene of hNGF was inserted into the KpnⅠ-BamHⅠ site of vector plasmid pSSHG-Neo to construct the vector of hNGF recombinant AAV. The recombinant AAV viral stock was packaged. Renal embryo 293 cell was co-transfected with the rAAV vector of plasmid pSSHG/hNGF, packaging plasmid pAAV/Ad and helper adenovirus pasmid pFG140 instead of adenovirus by calcium phosphate precipitation. Results The recombinant viral stock vector of plasmid pSSHG/hNGF was constructed successfully. The results of dot blot showed that we had obtained the rAAV stocks of high titre 1.46?10 12 PFU?mL -1. Conclusion We prepared the viral stock of rAAV-hNGF that can serve as the experimental study of gene therapy of SCI.

Objective To study the prokaryotic expression of fusion gene A?-HBcAg and analyze the immunoreactivity and immunogenicity of expression protein. Methods Recombinant plasmid pBV220/A?-HBcAg was transformed into E.coli DH5?, and expressed by temperature inducing. The bacteria were split by ultrasonic wave. The expression of the fusion protein was studied by SDS-PAGE and Coomassie brilliant blue staining. The immunoreactivity of the fusion protein was determined using ELISA. After immunized intraperitoneally with the fusion protein, 5 Balb/c mice's sera titers of anti- A? and anti-HBc were evaluated by ELISA. Results Fusion protein was in sediment of the split bacteria as inclusion bodies and its expression level was 5% of the total sediment protein. The fusion protein had both immunoreactivity of A? and HBcAg. The titers of anti-A? and anti-HBc were very low after 3 times of immunization. After immunization for 5 times, the titers reached 1∶800 and 1∶3 200 for anti-A? and anti-HBc, respectively. Conclusion Recombinant gene A?-HBcAg can be expressed in E.coli DH5? and the expression protein has certain immunoreactivity and immunogenicity. It indicates that further work should be done to enhance the expression level of fusion gene A?-HBcAg and improve the immunogenicity of the fusion protein.

Objective To prepare a hybridoma secreting stab le monoclonal antibodies against ?-amyloid peptide (A? 1-42) with high titer. Methods By genetic engineering technology, A ? gene was recombined with the MIR of HBcAg to get the A? and HBcAg fusi on protein. Spleen cells from BALB/c mice immunized with A? and HBcAg f usion protein were fused with mouse myeloma cells SP2/0. Results Two strains of hybridomas (1H 7 and 1F 3) secreting stable monoclonal antibodies raised against A? 1-42 were ob tained. The subtypes of A? 1-42 antibodies were IgG 3. C onclusion The A? 1-42 monoclonal antibodies obtained have high titers and specificity.