OBJECTIVE To investigate the pathogens and their drug-resistance in Fuwai Cardiovascular Hospital and provide antibiotics use suggestion for clinical treatment.METHODS The pathogens were identified by VITED 32 and analyzed by WHONET 5.4 RESULTS The pathogens mainly consisted of Gram-negative bacilli,which were highly sensitive to imipenem and meropenem except Pseudomonas aeruginosa;E.faecalis was much more sensitive to penicillin and gentamicin than E.faecium.Most coagulase-negative Staphylococcus(CNS) were resistant to oxacillin and showed low susceptibility rates to most antibiotics.No Gram-positive cooci were found to be resistant to vancomycin and teicoplanin.CONCLUSIONS To investigate the pathogens and their drug resistance is very important to prevent and control nosocomical infections.

OBJECTIVE To investigate the major pathogens and their antibiotics resistance in surgery wards in Fuwai Cardiovascular Hospital.METHODS The pathogens were classified and their antibiotics resistance was analyzed.RESULTS The pathogens mainly consisted of Gram-negative bacilli,which were sensitive to cefoperazone/sulbactam,pipercillin/tazobactam and amikacin with the sensitive rate of 62.7-97.6%.All Gram negative bacilli were sensitive to imipenem except Pseudomonas aeruginosa,with the sensitive rate of 95.2%~100%.In the Gram-positive bacteria,the susceptibility rates of Enterococcus faecium were lower than E.faecalis.Coagulase negative Staphylococcus and Staphylococcus aureus were mainly resistance to oxacillin and most other antibiotics,all Gram positive bacteria were sensitive to vancomycin.CONCLUSIONS To know the major pathogens and their susceptibility rates can prevent and control nosocomial infections effectively.

Objective:To establish an HPLC-ELSD method for the determination of mahuannin A in ephedrae radix et rhizoma. Methods:The content of mahuannin A was determined by an HPLC-ELSD method on an Alltima TM C18 column (250 mm × 4. 6 mm, 5 μm). The mobile phase was acetonitrile-water (28∶ 72) with a flow rate of 0. 7 ml·min-1, and the column temperature was 30℃. The temperature of drift tube heater was 105℃ and the flow rate of carrier gas was 2. 8 L·min-1 . Results:The linear range of mahua-nnin A was 42. 56-383. 04 μg·ml-1(r=0. 999 8). The average recovery and RSD was 99. 9% and 1. 96%(n=6), respectively. Conclusion:The method is simple and the result is accurate. It can be used for the quality control of ephedrae radix et rhizaoma.

Objective:To optimize the extraction process of the total flavonoids from Ephedrae Radix Et Rhizoma. Methods:The purification method of the total flavonoids from Ephedrae Radix Et Rhizoma was optimized with the yield and content of the total fla-vonoids as the indices. Based on the above research, the process parameters were optimized by an orthogonal test. Results:The opti-mum purification conditions were as follows:the volume fraction of ethanol was 50%, the stirring extraction time was 20 min, and the liquid-solid ratio was 8∶ 1(ml·g-1). Conclusion:The optimum purification technology is simple and reproducible, and suitable for the industrial production.

Objective:To study the fingerprint of Radix Salviae Miltiorrhizae of Xiangdan Injection. Methods : HPLC with UV detector was used to analyze the patterns of the Radix Salviae Miltiorrhizae of Xiangdan Injection. Protocatechuic aldehyde was used as internal standard substance. Results : The fingerprint of Radix Salviae Miltiorrhizae of Xiangdan Injection was set up and the fingerprint of them showed an excellent correlation. Conclusion : The study is helpful for the quality control of Xiangdan Injection.

To retrospective analyze the clinical profiles of 80 patients undergoing thoracotomy with protection of intercostal nerve versus traditional method.The doses of narcotics of two groups were (12 ± 5)and (43 ± 11) mg respectively.The postoperative levels of visual analogue score (VAS) and such potential complications as pneumonia,atelectasis and paraesthesia were examined (P ＜ 0.01).Protective technique of intercostal nerve during thoracotomy could effectively relieve postoperative chest pain,reduce the dosage of narcotics and lower the occurrence of lung complications.

Objective To discuss the clinical significance of anti-HBc-IgG(anti-HBc) only positivity of HBV markers in serum.Methods The total and the isolated anti-HBc positive rate were calculated retrospectively from 5 213 and 594 inpatients′ serum samples determined for five HBV markers including HBsAg,anti-HBs,HBeAg,anti-HBe and anti-HBc by microparticle enzyme immunoassay(MEIA),enzyme-linked immunosorbent assay(ELISA),respectively.The levels of anti-HBs were analysed in 167 five-HBV-marker negative and 124 anti-HBc-only positive subjects determined for five HBV markers by MEIA.Making a dilution with range from 2- to 30-fold with the phosphate buffer system containing 10% bovine calf serum to 97 serum samples with epidemiological anti-HBc only positivity screened from 594 inpatients determined for five HBV markers by ELISA,and anti-HBc of the dilute serum samples were determined by ELISA and MEIA,respectively.Results The total and the isolated anti-HBc positive rate with epidemiological and clinical significance by ELISA were 72.1%, 16.3 % and 62.6%,7.6%,respectively.The total and the isolated anti-HBc positive rate by MEIA were 78.1% and 13.2%,respectively.The difference of anti-HBs levels between five-HBV-marker negative and anti-HBc-only positive subjects determined for anti-HBs by MEIA was significant(?~2= 86.9 ,P

Objective To clone the human gene of Hepatitis B virus e antigen binding protein 1 (HBEBP1), which was screened with yeast two-hybrid system and bioinformatics techniques, to construct prokaryotic expression vector of pET-32a(+)-HBEBP1, and to induce the expression of recombinant protein in E. coli BL21. Methods The DNA fragment of HBEBP1 of about 372bp was amplified by reverse transcription PCR (RT-PCR), in which the mRNA was taken from HepG2 cells as the template, and cloned into pGEM-T vector. After restriction enzyme digestion identification and sequencing, the correct target DNA fragment was inserted into inducible prokaryotic expression vector pET-32a(+) and then transformed into competent E. coli BL21. By restriction enzyme digestion and PCR, the positive transformed clones were identified and induced with IPTG to obtain fusion protein. The HBEBP1 fusion protein was analyzed by Western blotting hybridization. Results The 372bp DNA fragment of HBEBP1 was amplified by RT-PCR. The recombinant expression vector pET-32a(+)-HBEBP1 was constructed successfully. After transformation with pET-32a(+)-HBEBP1 and induction with IPTG, the recombinant target protein of about 33kD was obtained, which was consistent with our anticipation. Western blotting assay showed that the protein had good specificity. Conclusions The recombinant prokaryotic expression vector pET-32a(+)-HBEBP1 is constructed, and the HBEBP1 gene is cloned successfully. The HBEBP1 fusion protein could be expressed in prokaryotic expression system of E. coli. These results lays a foundative for studying the immunogenicity and the biological characteristics of the HBEBP1 protein.

In order to screen genes transregulated by core protein of hepatitis C virus, cDNA microarray technology was employed to detect the gene expression change between HepG2 cells transfected with pcDNA3 1(-) core and the empty vector, respectively. Among 1152 genes, there were 95 genes with different expressions, of which 45 genes were upregulated and 50 genes were downregulated in HepG2 cells transfected with core protein expression plasmid. These genes transregulated by HCV core protein included human genes encoding proteins involved in cell proliferation, differentiation, apoptosis, signal transduction and immune regulation. Therefore, the results provided some new clues for further clarifying the molecular biology mechanism of pathogenesis and tumorigenesis of HCV core protein

To screen anti idiotypic single chain variable fragments(anti Id scFv)against hepatitis C virus NS4A(HCV NS4A)so as to lay a foundation for developing anti Id scFv vaccine againist the hepatitis C virus. The recombinant phage antibody library was panned by hepatitis C virus NS4A monoclonal antibody which was coated in a microwell plate. After five rounds of biopanning, 82 clones specific to HCV NS4A antibody were determined with the enzyme linked immunoadsorbent assay(ELISA). The specificity of anti idiotypic scFv was identified by ELISA and competitive inhibition assay. The DNA sequence of the positive clone was determined. The result showed that HCV NS4A anti Id scFv had a specific combination character with hepatitis C virus NS4A monoclonal antibody. The DNA sequence data showed that the anti Id scFv coding gene included 789 bp. The results suggested that the anti Id scFv fragments to HCV NS4A monoclonal antibody could be successfully selected by recombinant phage antibody library screening technique, which might pave a way for the study of new preventive and therapeutic strategy of hepatitis C using anti Id scFv.