BACKGROUND: With mechanical ventilation widely used in intensive care unit, the ventilator associated pneumonia (VAP) has become a common and serious complication in critically ill patients. Compared with adults, the incidence of VAP and the mortality are higher in children in pediatric intensive care unit (PICU) because of immune deficiency, severe basic diseases, and increased use of artificial airway or mechanical ventilation. Hence it is of significance to study the epidemiology and changes of antibacterial susceptibility in order to reduce the incidence and mortality of VAP in children. METHODS: From January 2008 to June 2010, 2758 children were treated in PICU of Wuhan Children's Hospital. Among them, 171 received mechanical ventilation over 48 hours in PICU, and 46 developed VAP. The distribution and drug-resistance pattern of the pathogenic bacteria isolated from lower respiratory tract aspirations were analyzed. RESULTS: A total of 119 pathogenic microbial strains were isolated. Gram-negative bacilli (G-) were the most (65.55％), followed by fungi (21.01％) and gram-positive cocci (G+, 13.45％). Among them, the most common pathogens were Acinetobacter baummannii, Escherichia coli, Klebsiella pneumoniae, candida albicans and coagulase-negative staphylococci. Antibiotic susceptibility tests indicated that the multiple drug-resistances of G- and G+ to antibiotics were serious. Most of G- was sensitive to ciprofloxacin, amikacin, imipenem, meropenem, cefoperazone-sulbactam and piperacillin-tazobactam. The susceptibility of G+ to vancomycin, teicoplanin and linezolid were 100％. Fungi were almost sensitive to all the antifungal agents. The primary pathogens of VAP were G-, and their multiple drug-resistances were serious. CONCLUSION: In clinical practice we should choose the most sensitive drug for VAP according to pathogenic test.

BACKGROUND: With beta-lactam drugs and immunosuppressants widely used, the infection caused byAcinetobacter baumannii (Ab) has become more and more serious with multidrug resistant Acinetobacter baumannii (MDRAb) emerging and worsening rapidly. Compared with other patients, the incidence and multidrug resistance of MDRAb are higher in children in pediatric intensive care unit (PICU) because of immune deficiency, severe basic diseases, prolonged hospitalization and invasive operations. Hence it is significant to study the epidemiology and changes of antibacterial susceptibility in order to reduce the incidence of MDRAb in children. METHODS: A total 115 patients with MDRAb pneumonia and 45 patients with negative MDRAb (NMDRAb) pneumonia who had been treated from January 2009 to August 2011 were studied retrospectively at the PICU of Wuhan Children's Hospital. Clinical data were analyzed with univariate and multivariate Logistic regression. RESULTS: In 176 clinical strains ofAcinetobacter baumannii isolated, there were 128 strains of MDRAb, accounting for 72.73％. Drug susceptibility tests showed that the resistance rates of β-lactam antibiotics were more than 70％ except for cefoperazone sulbactam. The rates to carbapenems were higher than 90％. They were significantly higher than those of NMDRAb. Amikacin, levofloxacin, ciprofloxacin and minocycline had the lowest drug-resistance rates (<20％). Multivariate Logistic regression revealed that ICU stay, the time of mechanical ventilation, anemia, hypoproteinemia and the use of carbapenems were independent risk factors for MDRAb pneumonia. CONCLUSIONS: MDRAb is an important opportunistic pathogen to pneumonia in PICU, and its drug-resistance is severe. It increases significantly the mortality of patients. It is important to take the effective prevention measures for controlling it.

In this paper, the five strains of Polygonatum odoratum were used as the experimental materials to test the supercooling point, freezing point, the degree of supercooling, the transition stage time, cooling time and water composition of the plant tissue. The cold resistance of P. odoratum was analyzed with the Gray Correlation Method. The results showed that the cold resistances of the five strains of P. odoratum were different, and the water content of plant tissue had some relevance with freezing point and supercooling point, whereas, it could not be measured when the moisture content was too low. The order of cold resistance of the five strains of P. odoratum was ZJCY, DYYZ, XYYZ, CYYZ and JZ I.
Cold Temperature ; Plant Roots ; chemistry ; physiology ; Polygonatum ; chemistry ; classification ; physiology ; Water ; analysis

A simple, rapid and sensitive method was developed for the quantification of tenofovir in plasma of Beagle dogs using HPLC-MS/MS analysis. The analytes tenofovir and internal standard (IS) adefovir were separated on a Zorbax SB-C18 column (3.5 microm, 100 mm x 2.1 mm, Agilent, USA) with mobile phase of methanol/water containing 0.3% formic acid using a gradient elution mode at a flow rate of 0.2 mL x min(-1). The plasma sample preparation was a simple deproteinization by the addition of 20% trichloroacetic acid followed by centrifugation. The detection was performed in positive selected reaction monitoring (SRM) mode with an electrospray ionization (ESI) source. The reactions monitored were m/z 288.1-176.2 for tenofovir and m/z 274.1-162.2 for adefovir (IS). Linear detection responses were obtained for tenofovir ranging from 10 to 5 000 ng x mL(-1). The intra- and inter-day precisions (RSD%) was no more than 6.3% with high recovery and good stability for the quantification, indicating the present method was specific, fast, accurate and reliable. The method was successfully applied to the pharmacokinetic study of two tenofovir agents. Tenofovir dipivoxil fumarate (BP0018, test agent) and tenofovir disoproxil fumarate (reference agent) were orally administrated to 8 Beagle dogs according to the 2 x 2 crossover design. Comparing with the reference agent, the longer MRT and t1/2 were obtained in the group of BP0018, while no significant difference was observed in AUC(0-t), AUC(0-infinity), C(max) and t(max) between them, suggesting that tenofovir dipivoxil fumarate was bioequivalent to the tenofovir disoproxil fumarate in Beagle dogs.
Adenine ; administration & dosage ; analogs & derivatives ; blood ; pharmacokinetics ; Administration, Oral ; Animals ; Area Under Curve ; Chromatography, High Pressure Liquid ; Dogs ; Female ; Male ; Organophosphonates ; administration & dosage ; blood ; pharmacokinetics ; Phosphorous Acids ; administration & dosage ; pharmacokinetics ; Reverse Transcriptase Inhibitors ; blood ; Tandem Mass Spectrometry ; Tenofovir

Effects of constituents from Schisandra chinensis (Wuweizi) on six liver microsomal CYP450 isozymes (CYP1A2, CYP2C6, CYP2C11, CYP2D2, CYP2E1 and CYP3A1/2) were studied in rats in vivo and in vitro. The in vitro incubation was conducted using liver microsomes of rats after multiple dosing of alcoholic/water extract from Schisandra chinensis. A HPLC-MS method was applied to determine the metabolites formation of six CYP450s probe substrates (phenacetin-CYP1A2, dextromethorphan-CYP2D2, diclofenac sodium-CYP2C6, mephenytoin-CYP2C11, chlorzoxazone-CYP2E1 and midazolam-CYP3A1/2) in rat liver microsomal incubations. The activity of CYP450 isozymes were represented by the formation of metabolites. Alcoholic extract of Schisandra chinensis (28-120 microg x mL(-1)) showed significant inhibitory effect on six CYP450 isozymes to a certain extent in vitro. Multiple dosing of Schisandra chinensis alcoholic extract (1.5 g x kg(-1), qd x 7d) had significant induction on CYP2E1 and CYP3A1/2, inhibition on CYP2D2 and CYP2C11, and no effect on CYP2C6 and CYP1A2. Water extract of Schisandra chinensis (100-500 microg x mL(-1)) also exhibited inhibition on the activity of CYP450 isozymes in vitro, whereas multiple administrations (1.5 g x kg(-1), qd x 7d) had significant induction of CYP2E1 and inhibition on CYP2D2, no effect on CYP2C6, CYP3A1/2, CYP1A2 or CYP2C11. The results suggested that the constituents from Schisandra chinensis exhibited the inhibition and induction on six rat liver microsomal CYP450 isozymes to a certain extent in vivo and in vitro. The possibility of interaction between Schisandra chinensis and coadministrative drugs will be considered base on the levels and subtype of CYP450 involved in the drug metabolism.
Animals ; Aryl Hydrocarbon Hydroxylases ; metabolism ; Cytochrome P-450 CYP1A2 ; metabolism ; Cytochrome P-450 CYP2E1 ; metabolism ; Cytochrome P-450 CYP3A ; metabolism ; Cytochrome P-450 Enzyme System ; metabolism ; Cytochrome P450 Family 2 ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Isoenzymes ; metabolism ; Lignans ; isolation & purification ; pharmacology ; Microsomes, Liver ; enzymology ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Schisandra ; chemistry ; Steroid 16-alpha-Hydroxylase ; metabolism ; Steroid 21-Hydroxylase ; metabolism ; Substrate Specificity

Adolescent ; Antibodies, Viral ; blood ; China ; epidemiology ; Female ; Humans ; Influenza A virus ; immunology ; Male ; Seroepidemiologic Studies ; Students, Medical ; Vietnam ; epidemiology ; Young Adult

OBJECTIVESlow transit constipation (STC) is a colonic motor disorder whose etiology remains unclear. Recent studies have demonstrated a crucial role for interstitial cells of Cajal (ICC) in regulation of intestinal motility. The aim of this study was to examine the distribution of ICC within the normal sigmoid colon and STC patients.

METHODSTwelve patients with STC and eight age-matched controls were studied. ICC were identified with a monoclonal antibody to c-kit by an indirect immunofluorescence method. Immunostained tissues were examined with a laser scanning confocal microscope and the area occupied by ICC was calculated with image analysis software.

RESULTSICC were located in the external muscle layers including longitudinal muscle (LM), myenteric plexus (MP), circular muscle (CM) and submucosal border (SMB). Two types of Kit-positive ICC were observed: bipolar cells characterized by one or two long processes, and multipolar cells with long stellate processes extending in various directions. A higher percentage of ICC was present in the MP regions and CM layers compared with the SMB and LM layers. Tissues from STC patients showed considerably decreased in number of ICC located in the four regions (ICC-LM, ICC-MP, ICC-CM, ICC-SMP), especially for ICC-SMP, almost completely disappeared.

CONCLUSIONDecreased c-kit + ICC in number may play an important role in the pathophysiology of STC. It remains to be determined whether loss of ICC is primary or secondary to another lesion.

Adult ; Aged ; Case-Control Studies ; Colon, Sigmoid ; pathology ; Constipation ; pathology ; physiopathology ; Female ; Fluorescent Antibody Technique, Indirect ; Gastrointestinal Transit ; physiology ; Humans ; Male ; Middle Aged

OBJECTIVETo explore the influence of methylprednisolone (MP) on cellular component in donor graft and on H-2 haploidentical hematopoietic stem cell transplantation (HSCT) in mice.

METHODSA murine model of H-2 haploidentical HSCT was established by using of c57BL/6J male mouse as donor and (c57BL/6J x LB/C) F1 female mouse as recipient. The donor mouse received peripheral-blood (PB) progenitor cells mobilization regimens consisted of recombinant human granulocyte colony-stimulating factor (rhGCSF) alone (control group) or combined with MP in dose of 2 mg/kg daily [small-dose (SD) group], 10 mg/kg daily [middle-dose (MD) group], and 50 mg/kg daily [large-dose (LD) group] respectively. Percentage of T cell subsets, DC1 (HLA-DR+CD11c+) and CD34+ cell in the grafts were detected by flow cytometry. Transplant rejection,severity of GVHD and survival time were observed.

RESULTSThe percentages of CD3+ T cell in donor grafts in the three groups were significantly lower than that in control group (P < 0.05). The percentage of CD3+ CD4+ T cells decreased more significantly than that of CD3+ CD8+ T cells, and CD4/CD8 ratios decreased significantly. The percentage of CD4+ CD25+ T cells increased significantly, the percentage of DC1( HLA-DR+CD11c+) decreased and the percentage of CD34+ cells increased in all the three groups than in control group. There were significant differences in the percentage of CD3+ T cells, CD3+ CD4+ T cells and CD34+ cells in donor grafts among SD group, MD group and LD group (P < 0.05). The engraftment rates in control, SD, MD and LD groups were 90%, 100%, 100% and 80% respectively. Severity of aGVHD in each study group decreased significantly compared with that in control group (P < 0.05). There were statistical differences among different dosage groups (P < 0.05). Survival time after transplantation in all study groups were significantly longer than that in control group (P < 0.05), and in MD group was significantly longer than in SD group and LD groups (P < 0.05).

CONCLUSIONSAddition of methylprednisolone to routine donor mice HSC mobilization regimen has a definite effect in alleviating aGVHD and prolonging survival time of mouse after H-2 haploidentical HSCT. With a suitable dosage addition of methylprednisolone to donor mice HSC mobilization regimen could avoid the increasing risk of graft rejection.

Animals ; Antigens, CD34 ; Dendritic Cells ; drug effects ; immunology ; Female ; Graft Rejection ; prevention & control ; Graft vs Host Disease ; prevention & control ; Hematopoietic Stem Cell Mobilization ; Hematopoietic Stem Cell Transplantation ; Male ; Methylprednisolone ; administration & dosage ; Mice ; Mice, Inbred C57BL ; T-Lymphocyte Subsets ; drug effects ; immunology

OBJECTIVETo investigate the variance of antibiotic resistance of Acinetobacter Baumanii isolated from children in Wuhan between 2006 and 2008.

METHODSBacterial susceptibility testing was carried out by the Kirby-Bauer method in 679 strains of Acinetobacter Baumanii isolated in Wuhan Children's Hospital between 2006 and 2008. The results were assessed according to the guidelines of the Clinical and Laboratory Standards Institute (2008).

RESULTSThe nonsusceptible rates of Acinetobacter Baumanii to ceftazidime, cefepime and piperacillin/sulbactam increased significantly in 2007 compared with those in 2006 (P<0.05). By comparing the results of 2007, it was suggested that the nonsusceptible rates of Acinetobacter Baumanii to ceftazidime, cefepime, piperacillin/sulbactam, cefoperazone/sulbactam, imipenem and meropenem increased significantly in 2008 (P < 0.05).

CONCLUSIONSThe nonsusceptible rates of Acinetobacter Baumanii to beta-lactam antibiotics in children from Wuhan increased significantly year by year between 2006 and 2008.

Acinetobacter baumannii ; drug effects ; Anti-Bacterial Agents ; pharmacology ; Child ; Drug Resistance, Bacterial ; Humans ; Microbial Sensitivity Tests ; Time Factors

OBJECTIVETo compare the outcomes of intracytoplasmic sperm injection (ICSI) with retrieved epididymal and testicular sperm for obstructive azoospermia and with ejaculated sperm for severe oligozoospermia and asthenospermia.

METHODSWe retrospectively analyzed 431 ICSI cycles, which were divided according to sperm sources into Groups A (n=287 in patients with severe oligozoospermia or asthenospermia using ejaculated sperm), B (n=109 in obstructive azoospermia patients with sperm retrieved by percutaneous epididymal sperm aspiration, PESA) and C (n=35 in obstructive azoospermia patients with sperm retrieved by testicular sperm extraction, TESE). Comparisons were made among the three groups in the rates of embryo implantation, fertilization, pregnancy, cleavage, and miscarriage.

RESULTSGroup A showed statistically significant differences from Groups B and C in the rates of embryo implantation and pregnancy (18.46% vs. 25.23% and 28.76%, 31.23% vs. 42.16% and 39.39%, P < 0.05). But no significant differences were seen in the rates of fertilization, cleavage and miscarriage among the three groups (P > 0.05).

CONCLUSIONThe rates of embryo implantation and clinical pregnancy are higher in patients with obstructive azoospermia than in those with severe oligozoospermia or asthenospermia after ICSI with ejaculated sperm.

Azoospermia ; therapy ; Epididymis ; cytology ; physiopathology ; Female ; Humans ; Male ; Oligospermia ; therapy ; Pregnancy ; Pregnancy Outcome ; Retrospective Studies ; Sperm Injections, Intracytoplasmic ; methods ; Spermatozoa ; Testis ; cytology ; physiopathology