Adolescent ; Diagnosis, Differential ; Endometriosis ; pathology ; Female ; Humans ; Melanoma ; pathology ; Melanosis ; complications ; pathology ; surgery ; Ovarian Neoplasms ; complications ; Peritoneal Diseases ; complications ; pathology ; surgery ; Teratoma ; complications

Objective To investigate the distribution and drug susceptibility of vaginal pathogens in pregnant women during perinatal period.Methods Vaginal discharge specimens of 10 800 women (5 400 were pregnant women during perinatal period,5 400 were non-pregnant women) were performed bacterial culture and drug susceptibility testing.Results The isolation rate of pathogens from 5 400 perinatal pregnant women was 26.00％ (n =1 404),including 759 strains of fungi,611 strains of gram positive cocci,32 strains of gram-negative bacilli,2 strains of Neisseria gonorrhoeae;among 5 400 non-pregnant women,the isolation rate of pathogens was 7.87％ (n=425),including 232 strains of fungi,182 strains of gram-positive cocci,5 strains of gram negative bacilli,and 6 strains of Neisseria gonorrhoeae.Resistance rates of Streptococcus agalactiae isolated from perinatal pregnant women to erythromycin and clindamycin were 84.85％ and 80.81 ％ respectively,resistance rate of Staphylococcus aureus to erythromycin was 40.91 ％,resistance rates of Escherichia coli to tetracycline and sulfamethoxazole/trimethoprim were 69.23 ％ and 53.85 ％ respectively,resistance rate of Neisseria gonorrhoeae to penicillin and sulfamethoxazole/trimethoprim was 100.00％.Conclusion Vaginal infection rate in perinatal pregnant women is higher than non-pregnant women,screening of vaginal pathogens in perinatal pregnant women should be strengthened,so as to ensure the safety of mothers and infants.

Objective To understand the clinical characteristics of pediatric patients who developed H1N1 influenza A virus-associated pneumonia during the outbreak of H1N1 influenza A in Shanghai. MethodsA dcscriptivc study was done to analyze the clinical and epidemiologic characteristics of 30 hospitalized children who developed complicated pneumonia caused by H1N1 influenza A virus infection in 2009 in Shanghai. The comparison of medians was done using rank sum test and comparison of rates was done using exact chi-square test. Results Among thirty pediatric patients with H1N1 influenza A virus-associated pneumonia, the median age was 5.9 years old, five cases (16.7 %) had pre-existing medical conditions. Twenty cases (66.7 % ) had been exposed to the classmates or family membcrs with fever. All cases had fever and cough. Eleven cases (36.7 %00 ) had tachypnca and ten (33.3%) had wheeze. Eleven cases (36.7%) showed white blood cell (WBC)＜4.0 × 109/L and 2 (6. 7%) had thrombocytopenia. All patients had bilateral or unilatcral patchy infiltrates in the lung indicated by chest X-ray and four (13. 3%) had extensive infiltrates with the evidence of pulmonary edema. One (3. 3%) critically ill child with pneumonia, chest computed tomography scan revealed lung fibrosis 3 months and 9 months after illness onset. Three(10. 0%) cases had pneumomediastinum and subcutaneous emphysema. Six cases (20. 0%) were complicated with acute respiratory failure, three (10. 0%) with acute asthmatic attack and one (3. 3%) with encephalitis. All patients were treated with oseltamivir plus antibiotics and four required mechanical ventilation. All patients survived. The median duration of fever in group with oseltamivir given within 2 days of fever onset was statistically shorter than that in group with oseltamivir given 2 days after fever onset (2 days vs 5 days, Z= -8. 015, P<0. 01). Conclusions Both pre-school age and schoolage children may develop complicated severe respiratory diseases after H1N1 influenza A virus infection. Early initiation of oseltamivir may shorten the duration of fever and reduce the occurrence of severe complications.

Objective To assess the academic level of papers on systematic reviews and meta-analysis published in Chinese Journal of Pediatricsand methodology they used.Methods Basic data were extracted from 13 papers on sys-tematic reviews and meta-analysis published in Chinese Journal of Pediatrics .The methodology they used was assessed according to the preferred reporting items for systematic reviews and meta-analysis (PRISMA) and ANSTAR Scale and analyzed using the RevMan5.0.Results The PRISMA score was 14-23.5 (mean 20.0±3.11) and the AMSTAR score was 3-7.5 (mean 6.04±1.38) for the methodology used in papers on systematic reviews and meta-analysis published in Chinese Journal of Pediatrics .Conclusion The methodology used in papers on systematic reviews and meta-analy-sis published in Chinese Journal of Pediatrics is not quite valid and should thus be improved .

OBJECTIVE: To evaluate enantiomeric separation methods for beta-blocking agents and analogs. METHODS: Enantiomeric separation of racemates of 11 beta-blocking agents and their analogs was performed using chiral stationary phases and 2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosyl isothiocyanate (GITC). RESULTS: These beta -blocker racemates were separated into enantiomers in one or several chormatographic states such as propranolol, bisoprolol, metoprolol, celiprolol, carvedilol, sotalol, propafenone, ephedrine, and zomitriptan. Temperature had a significant effect on the resolution of the drugs when using chiralcel OD. Lower temperatures were associated with higher resolutions. CONCLUSION: When separating beta-blocking agents and their analogs, Chiralcel OD, Chiralpak AD, Chiral stationary phases and GITC chiral derivative reagents have complementary functions.

OBJECTIVETo obtain recombinant human CYP2E1 and to determine its activity by using the specific probe substrate.

METHODSCYP2E1 cDNA was obtained by RT-PCR using human liver RNA as template. The cloned CYP2E1 cDNA was ligated with pFastBac vector to generate recombinant pFastBac-CYP2E1, which was then transformed into E. coli DH 10 Bac. Recombinant Bacmid-CYP2E1 was generated by transposition. Then Spodoptera frugiperda (Sf9) insect cells was infected with Bacmid-CYP2E1 to generate recombinant baculoviruses carrying human CYP2E1 cDNA. Finally, Sf9 insect cells were triinfected with recombinant baculoviruses carrying human CYP2E1, CYPOR and CYPb5. The activity of the recombinant enzymes was determined using chlorzoxazone as the substrate.

RESULTThe Kmand Vmaxof recombinant CYP2E1 to chlorzoxazone was (72.4 +/-8.7) micromol. L(-1) and (2.41 +/-0.10) micromol.min(-1)?g(-1)protein, respectively.

CONCLUSIONActive recombinant CYP2E1 has been obtained by bac-to-bac expression system and its activity is similar to previous reports.

Animals ; Baculoviridae ; genetics ; metabolism ; Cytochrome P-450 CYP2E1 ; biosynthesis ; genetics ; Escherichia coli ; genetics ; Genetic Vectors ; genetics ; Humans ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Spodoptera ; genetics ; metabolism ; Transfection

OBJECTIVETo develop a RP-HPLC method for the determination of quercetin in UGT1A3 cDNA-transfected cells.

METHODSThe lysate of cells transfected with human recombinant uridine 5-diphosphate glucuronosyltransferases UGT1A3 cDNA was co-incubated with quercetin, the reaction was terminated with acetonitrile, and luteolin was used as internal standard. The determination was performed on a C(1) reversed phase column with a mobile phase of methanol-0.1% formic acid (V/V) at a flow rate of 1.0 ml/min. The gradient elution was as follows: 0 - 25 min (30:70-80:20, methanol:0.1% formic acid), > 25-25.5 min (80:20), >25.5-27 min (80:20-30:70), > 27-30 min (30:70). A UV-VIS detector was operated at 368 nm.

RESULTThe standard curve was linear over the concentration range of 5-200 μmol/L (r = 0.9999). The limit of detection was 1.25 μmol/L(S/N ≥ 3), and the limit of quantification was 5 μmol/L (S/N >10, RSD = 6.99%). The method afforded recoveries of 99.1%-103.5%, and precisions for inter- and intra-assay were < 2.5% and < 8%, respectively. In addition, kinetic analysis indicated that the K(m), V(max) and CL(int) (V(max)/K(m)) values for quercetin glucuronide were (62.95 ± 13.16) μ mol/L, (284.50 ± 24.35)nmol*min⁻¹*g⁻¹ and 4.52 ml*min⁻¹*g⁻¹, respectively.

CONCLUSIONThe method established is accurate and simple and suitable for the determination of quercetin in UGT1A3 cDNA-expressed cells.

Cells, Cultured ; Chromatography, High Pressure Liquid ; methods ; Glucuronosyltransferase ; genetics ; Humans ; Quercetin ; analysis ; pharmacokinetics ; Transfection

OBJECTIVETo investigate the biological and clinical features of Chinese rhesus monkeys after intravenous (IV) and intrarectal (IR) challenge with SIVmac239 in rhesus monkeys of Chinese origin, and compare the differences between the routes of infection.

METHODSRhesus monkeys of Chinese origin were inoculated with SIVmac239 either by IV (n = 19) or IR (n = 6) routes. Simian immunodeficiency virus (SIV)-specific antibody titer, CD4 + T cell counting, plasma SIV load, lymph node pathology, and clinical manifestations were compared between these two groups 232 or 168 days after challenging.

RESULTSAll SIVmac239-inoculated animals became seropositive for SIV-specific antibodies. SIV-specific IgM was detected in IV groups as from day 10 but was not detected in IR for all the time points. Although SIV-specific IgG was detected as from day 30 in both groups, the IgG titers were ten-fold higher in IV group than in IR group after day 168. CD4 + T-cell counting decreased progressively in IV group but remained stable in IR group over time. Plasma SIV RNA loads peaked in all animals between day 10 and day 14 (10(7) copies/ml), then declined to "setpoint" (10(3) - 10(6) copies/ml) about 2 months later. Most inoculated animals manifested lymphadenopathy. Two animals in IV group and one in IR group died of simian AIDS between day 150 and day 210, as evidenced by the autopsies showing the depletion of lymph tissues, Pneumocystis carinii pneumonia and other opportunity infections. Conclusion IV or IR inoculation of SIVmac239 in Chinese rhesus monkeys will result in chronic SIV infection with a similar clinical feature of natural HIV infection, which provides an excellent experimental animal model for AIDS.

Animals ; CD4-Positive T-Lymphocytes ; metabolism ; China ; Disease Models, Animal ; Female ; Macaca mulatta ; virology ; Male ; Rectum ; virology ; Simian Acquired Immunodeficiency Syndrome ; immunology ; virology ; Simian Immunodeficiency Virus ; immunology ; pathogenicity ; Veins ; virology

CYP2D6 is an important drug-metabolizing enzyme. The polymorphism of CYP2D6 leads to metabolism difference and the different reactions of drugs in the individuals and different races are normal phenomenon in clinical medication. CYP2D6*10 is an important subtype in Asian people and 51.3% Chinese are classified with this subtype. To obtain recombinant active CYP2D6*1/CYP2D6*10 in baculovirus system by optimizing coexpression with CYPOR, and detect their activity to catalyze dextromethorphan, three recombinants pFastBac-CYP2D6*1, pFastBac-CYP2D6*10 and pFastBac-CYPOR were constructed and transformed into DH10Bac cell to obtain the recombinant Bacmid-CYPOR, Bacmid-CYP2D6*1 and Bacmid-CYP2D6*10. And then the recombinant CYP2D6*1 and CYP2D6*10 virus were obtained by transfecting Sf9. Then homogenate protein activity was determined with dextromethorphan as substrate. The multiple of infection (MOI) and its ratio of recombinant CYP2D6 virus to CYPOR virus were adjusted by detecting the activity of the homogenate protein. The Km and Vmax are 26.67 +/- 2.71 micromol x L(-1) (n=3) and 666.7 +/- 56.78 pmol x nmol(-1) (CYP2D6) x min(-1) (n=3) for CYP2D6*1 to catalyze dextromethaphan. The Km and Vmax are 111.36 +/- 10.89 micromol x L(-1) (n=3) and 222.2 +/- 20.12 pmol x nmol(-1) (CYP2D6) x min(-1) (n=3) for CYP2D6*10 to catalyze dextromethorphan. There is significant difference between CYP2D6*1 and CYP2D6*10 for Vmax and Km (P < 0.01). The clearance ratio of CYP2D6*1 is 25.0 and the clearance ratio of CYP2D6*10 is 2.0. The expressed CYP2D6*1 and CYP2D6*10 are useful tools to screen the metabolism profile of many xenobiotics and endobiotics in vitro, which are benefit to understand individual metabolism difference.
Animals ; Baculoviridae ; enzymology ; genetics ; Catalysis ; Cells, Cultured ; Chromatography, High Pressure Liquid ; methods ; Cytochrome P-450 CYP2D6 ; genetics ; metabolism ; Dextromethorphan ; metabolism ; Isoenzymes ; metabolism ; NADPH-Ferrihemoprotein Reductase ; genetics ; metabolism ; Plasmids ; Recombinant Proteins ; genetics ; metabolism ; Spectrometry, Mass, Electrospray Ionization ; Spodoptera ; cytology ; virology ; Transfection