20 antimicrobial agents were determined by Kirby-Bauer methods by the participating institutions.RESULTS A total of 428 strains were isolated,Gram-positive cocci and Gram-negative bacilli accounted for 49.8% and 50.2%,respectively.The Gram-positive bacteria increased from 44.6% to 51.2% from 1995-1996 through 2003-2004 and Gram negative bacteria decreased from 55.4% to 48.2% in the meantime.The most frequent Gram-positive isolates were coagulase negative staphylococci,Staphylococcus aureus,Enterococcus spp and Streptococcus pneumoniae;the most frequent Gram-negative isolates were Acinetobacter spp,Klebsiella spp,Escherichia coli,Pseudomonas aeruginosa and Enterobacter spp;Neisseria meningitidis,Haemophilus influenzae,Flavobacterium spp and Citrobacter spp were relative less common.No strains resistant to vancomycin were found in staphylococci and enterococci.Gram-negative bacilli were highly susceptible to carbapenem.CONCLUSIONS Gram-positive cocci play an increasing role in central nervous sysytem infections,especilly coagulase negative staphylococci,and Gram-positive cocci have been increasing from 1995-1996 through 2003-2004,resistance to the first line antibiotics is common among all cerebrospinal fluids isolates.

Objective To discuss effects of anti-inflammatory mechanism of amygdalin on rats with type II collagen-induced arthritis (CIA). Methods Wistar rats were randomized into normal group, model group, amygdalin group, and tripterygium group. Type II CIA rat models were established. From the 15th day after the modeling establishment, each administration group was given corresponding dose of medicine for continuous 28 days. Levels of TNF-αand sICAM-1 were detected by ELISA in serum of rats, and expression of TNF-α was detected by immuno-histochemical method. Results TNF-α positive expression in amygdalin group and tripterygium group was similar and significantly reduced compared with model group. Levels TNF-α and sICAM-1 in amygdalin group and tripterygium group significantly decreased compared with those in model group (P<0.05), without significant difference compared with normal group (P>0.05). Conclusion Amygdalin can inhibit the expression of TNF-α and levels of TNF-α and sICAM-1, in order to treat rheumatoid arthritis.

Objective To observe the effect of Danzhi Jiangtang Capsules(DJC) on serum interleukin-18 (IL-18),cystatin C (CysC) and relative biochemical indexes in patients with early diabetic kidney disease(DKD),and to explore its protective action on the kidney.Methods Sixty-two hospitalized patients with early diabetic nephropathy (period 11-Ⅲ) in Endocrinology Department of the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine were divided into western medicine group and DJC group,31 cases in each group.The patients in western medicine group received conventional hypoglycemic drugs,and DJC group received DJC based on the treatment for the western medicine group.Before and after treatment,we observed the levels of serum IL-18,CysC,fasting plasma glucose (FPG),2-hour postprandial glucose (2hPG),glycosylated hemoglobin (HbA1c),serum creatinine (SCr),blood urea nitrogen (BUN),microalbumin (mAlb),urinary albumin /creatinine ratio (UACR),and estimated glomerular filtration rate (eGFR).Results The total effective rate was 90.0％ in DJC group and 76.7％ in the western medicine group,there being significant difference between the two groups(P ＜ 0.05).After treatment,the main traditional Chinese medical syndrome scores of DJC group were significantly lower than those before treatment,and the improvement was better than that of the western medicine group (P ＜ 0.05 or P ＜ 0.01).After treatment,the serum levels of IL-18,CysC,2hPG,SCr,BUN,mAlb,UACR in the two groups were significantly lower than those before treatment (P ＜ 0.05 or P ＜ 0.01),while eGFR level was higher than that before treatment (P ＜ 0.05 or P ＜ 0.01),and the improvement effect (except for BUN) in the DJC group was superior to that in the western medicine group(P ＜ 0.01).DJC had an effect on lowering the FPG and HbA1c (P ＜ 0.05 or P ＜ 0.01 compared with those before treatment),and the western medicine had no obvious effect on the two indexes (P ＞ 0.05).The results of Spearman correlation analysis showed that IL-18 was correlated with CysC,IL-18 and CysC were correlated with HbA1c,mALB and uACR.Conclusion DJC can protect renal function of patients with early diabetic nephropathy.

Objective To investigate the different concentrations of progesterone on the expression of a disintegrin and metalloprotease 10 (ADAM10),long form leptin receptor (Ob-R) and the secretion of soluble leptin receptor (SLR),leptin (LEP) in primary pregnancy human chorionic trophoblast cells. Methods Cultured primary human trophoblast cells and added in different concentrations of progesterone (0,100,150 and 200 ng/mL) for 24 hours. The relative expression of ADAM10 and Ob-R in the cells and the content of SLR and LEP in the supernatant were detected. Results With increasing concentrations of progesterone,early human trophoblast cells ADAM10 content gradual-ly decreased,the difference between the two groups was significant (P<0. 05). With increasing concentrations of progesterone,human chori-onic trophoblast cells in early pregnancy Ob-R expression levels increased, the difference was no statistically significant between each two groups (P>0. 05). SLR content of the cell supernatants as the concentration of progesterone increased and decreased,there are significant differences between each two groups (P>0. 05). LEP cell supernatant in each group with the increase of the concentration of progesterone concentration increased gradually between the two groups were significantly different(P>0. 05). Pearson’s test showed that the expression of expression SLR and LEP exists a significant negative correlation (R= -0. 949,P<0. 01). Conclusion Progesterone may influence the ex-pression of ADAM10,SLR and LEP by the regulation of leptin to participate GDM occurs.

OBJECTIVE To characterize clinical feature,frequency of isolation and antimicrobial susceptibility of bacterial pathogens isolated from patients with healthcare-associated bacterial meningitis(HABM).METHODS We review the charts of all patients in whom the diagnosis was based on(national) diagnostic criteria of healthcare-(associated) infections at Huashan Hospital from 1995 through 2004.The pathogens were routinely isolated,(identified) and susceptibilities against antimicrobial agents were determined by Kirby-Bauer methods.RESULTS During the 10-year study period,109 patients were treated for 120 episodes of HABM.Most of patients had a(history) of recent and remote neurosurgery.Fever was present in all patients,while nuchal rigidity and decrease consciousness were present in less than half of all patients.CSF opening(pressure,) white blood cell count and(protein) were elevated with predominance of neutrophils.A total of 120 strains were isolated from CSF specimen,Gram-positive bacteria and Gram-negative bacilli were accounted for 35.8% and(64.2%) of all isolates,respectively.Acinetobacter spp(24.2%),coagulase-negative staphylococci(22.5%),Klebsiella pneumoniae(12.5%),Pseudomonas aeruginosa(10%),Enterobacter cloacae(8.3%) and Staphylococcus aureas(7.5%) were the 6 most frequent isolates and resistance to the first line antibiotics was common among all pathogens isolated.(CONCLUSIONS) The most common risk factor for HABM is neurosurgery.Gram-negative bacilli and staphylococci are important causes of HABM,resistance to the first line antibiotics is common among all pathogens isolated.

Objective: To explore influence of sleep deprivation (SDP) on serum interleukin-1β (IL-1β) level in rats with coronary atherosclerosis (CAS). Methods: A total of 96 healthy SD rats were randomly and equally divided into normal control group, CAS group, SDP group and SDP + CAS group according to number table; rats in CAS group and SDP + CAS group were fed with high cholesterol diet to establish CAS animal model; SDP group and SDP + CAS group received SDP through small platform with water environment. Radioimmunoassay was used to measure serum concentration of IL-1β; and serum concentrations of total cholesterol (TC), triglyceride (TG), high density lipoprotein-cholesterol (HDL-C) and low density lipoprotein-cholesterol (LDL-C) were measured respectively; above indexes were compared among four groups. Results: (1) Compared with serum IL-1β level of normal control group, there were significant increase in SDP group, CAS group and SDP + CAS group[(13.33±3.20) ng/L vs. (14.40±4.41) ng/L vs. (22.21±5.94) ng/L vs. (30.17±7.45) ng/L] (P<0.05～<0.01); (2) Levels of TC, TG and LDL-C in CAS group and SDP + CAS group were significantly higher, level of HDL were significantly less than those of normal control group and SDP group (P<0.05～<0.01), levels of above indexes in SDP + CAS group were significantly higher or less than those of CAS group (P<0.05 all). Conclusion: Sleep deprivation can significantly heighten serum interleukin-1β level and total cholesterol, triglyceride, low density lipoprotein-cholesterol levels, decrease HDL level in rats with coronary atherosclerosis or sleep deprivation.

Objective To investigate the molecular characteristic,the virulence factors and antimicrobial resistance of Methicillin-resistant Staphylococcus aureus (MRSA) isolated from pediatric patients.Methods Ninety-eight non-duplicate strains of and 49 non-duplicate strains of Methicillinsusceptible Staphylococcus aureus (MSSA) isolated from the three children's hospitals in Shanghai in 2008 were investigated.Panton-valentine leukocidin (PVL) gene was detected by polymerase chain reaction (PCR).The genotypes of staphylococcal cassette chromosome mec (SCCmec) of the MRSA isolates were confirmed by multiplex PCR.The sequence type (ST) of each strain was determined by multilocus sequence typing (MLST),and the algorithm eBURST was used to identify groups of clonal complex (CC).The minimal inhibitory concentrations (MIC) of fourteen antibiotics for all isolates were determined by agar dilution method.Results Among 98 isolates of MRSA,the positive rate of PVL genes was 6.1％ (6/98).In contrast,the positive rate of PVL genes was 4.1％ (2/48) of the MSSA strains.Among 98 isolates of MRSA,4.1％ (4/98),23.5％ (23/98),53.0％ (52/98) and 15.3％ (15/98) of the strains harboured SCCmec types Ⅱ,Ⅲ,Ⅳ and Ⅴ,respectively. The remaining four isolates (4.1 ％) presented a unique SCCmec pattern that could not be classified to any known types by the employed typing assays.Combining the ST and SCCmec type,the predominant clones were ST59-SCCmec Ⅳ (30 strains) and ST239-SCCmec Ⅲ (23 strains),followed by ST5-SCCmecⅣ and ST1-SCCmecⅣ (8 strains for each clone),ST239-SCCmec Ⅴ (6 strains),ST88-SCCmecⅤ (5 strains),ST5 SCCmecⅡ (4 strains),ST59-SCCmec Ⅴ (3 strains),ST8-SCCmecⅣ and ST88-SCCmecⅣ (2 strains for each clone),ST22-SCCmecⅣ,ST910-SCCmecⅣ and S45-SCCmec Ⅴ (1 strain for each clone),eBURST analysis distributed the MRSA isolates into several CC.ST8 and ST239 belonged to ST8 CC,ST1 belonged to ST15 CC,ST910 belonged to ST 30 CC,ST59,ST5,ST88,ST45,ST22,ST9 and ST7 were the origin of their own CC.The results of MIC showed that the 67 strains of MRSA harboring SCCmec type Ⅳ or SCCmec type Ⅴ were more susceptible to various non-β-lactam antibiotics than 27 strains of MRSA harboring SCCmec type Ⅱ or SCCmec type Ⅲ,and no vancomycin-resistant strain was found.Conclusions In three children's hospitals in Shanghai,the PVL gene-positive rate of MRSA isolates is relatively low,SCCmec type Ⅳ and SCCmec type Ⅴ could spread among hospitals to cause a small scale epidemic and have a variety of ST.

Objective To learn the current in vitro antimicrobial susceptibility of Mycoplasma pneu-moniae in Shanghai and to understand the mechanisms of resistance to macrolides. Methods M. pneumoniae was isolated from pediatric patients with low respiratory tract infections(RTI) using broth and PPLO agar medi-um. PCR amplification and sequence analysis of P1 adhesion gene were performed to identify all M. pneumoniae strains. Susceptibility testing was carried out for macrolides, tetracyclines and fluoroquinolones using broth mi-crodilution method with SP4 broth. PCR amplification and sequence analysis of 23S rRNA genes were performed for all M. pneumoniae strains. P1 gene PCR-RFLP typing was performed to subtype the M. pneumoniae strains. Results One hundred and two M. pneumoniae strains were isolated in Shanghai from Oct 2005 to Dec 2008. All M. pneumoniae isolates were susceptible to the tetracyclines and fluoroquinolones tested. Of 102 clinical isolates, 83(81.4%) was resistant to erytbromycin and all 83 erythromycin-resistant strains had MIC>128 mg/L. An increasing trend of resistance rates were showed: 16.7% (1/6) in 2005, 76.5% (13/17) in 2006, 100.0% (24/24) in 2007 and 81.8% (45/55) in 2008. All macrolide-resistant M. pneumoniae strains harbored an A2063G transition mutation in domain V of 23S rRNA genes. The P1 gene RFLP type 1 is predominant (85.3%, 87/102) in M. pneumoniae clinical isolates. Conclusion The macrolide resistance rate of M. pneu-moniae is very high in Shanghai. The mechanism of macrolide resistance is associated with transition mutation on the 23S rRNA gene.

This study chiefly aimed at investigating the effects of Dan Zhi Jiang Tang Capsule (DJC) on myocardial Toll-like receptor 4 (TLR4),tumor necrosis factor α (TNFα) and interleukin-8 (IL-8) and the cardiopathologic changes in diabetic rats.Fifty male rats,excluding 8 rats in the control group,had been administered with high fat forage for 4 weeks,which established the diabetic rat model combined with the intraperitoneal injection of streptozotocin (STZ) at 35 mg· kg-1.The established model rats were randomly divided into the model group,the DJC high-dose group,the DJC low-dose group and the pioglitazone group.The rats in the model group and the control group were intragastrically administered with DJC at 1.08 and 0.54 g· kg-1 each day,while those of the pioglitazone group were treated with pioglitazone at 10 mg· kg-1 each day,and those of the model group and control group with saline of the same volume.The administrations lasted 8 weeks with the continues high-fat diet.The levels of TLR4,TNFα and IL-8 protein in myocardium of the rats were determined,and so were the blood glucose,HbA1c and blood lipids.Compared with the control group,the expressions of myocardial TLR4,TNFα and IL-8 in the model group were significantly increased (P＜ 0.01).The expressions of TLR4,TNFα and IL-8 in the DJC high-dose and low-dose groups were significantly decreased (P＜0.05 or P＜0.01).The cardiopathological report showed severe myocardial lesion in the model group.However,myocardial lesion in the DJC high-dose group was significantly mitigated,which presented better therapeutic effects than the DJC low-dose group and the pioglitazone group.The levels of blood glucose,HbA1c,TG,TC,LDL-C in the model group were significantly higher than those in the control group (P＜0.05 or P＜0.01).The levels of blood glucose,HbA1c,TG,TC and LDL-C were significantly decreased in the DJC high-dose and low-dose groups in comparison with the model group (P＜ 0.05 or P＜ 0.01).In conclusion,DJC down-regulated the expressions of TLR4,TNFα and IL-8 and improved the degree of myocardial lesion in the diabetic rats,showing favorable hypoglycemic and lipid-lowering effects.

Objective To develop a method for rapid detection of Mycoplasma pneumoniae and its macrolide resistance mutation. Methods The primers and cycling probe sets were designed to detect two single nucleotide mutation, A2063G and A2064G, in the 23s rRNA gene of Mycoplasma pneumoniae. By using recombinant plasmids containing 23s rRNA gene fragments, 102 Mycoplasma pneumoniae clinical isolates from 2005 to 2008, and 136 nasopharyngeal suction specimens from pediatric patients with low respiratory tract infections in Shanghai Children's Hospital from November to December in 2009 were investigated to determine the specificity and the sensitivity of the CycleavePCR method. PCR amplification and sequence analysis of 23S rRNA genes were performed for all Mycoplasma pneumoniae strains and Mycoplasma pneumoniae positive specimens to confirm the results of the CycleavePCR method. Results Of 102 clinical isolates, 83 was resistant to erythromycin and sequence results show that all macrolide-resistant Mycoplasma pneumoniae strains harbored an A2063G ( 82/83 ) or A2064G ( 1/83 ) transition mutation in 23S rRNA genes. Twelve was Mycoplasma pneumoniae detected positive by CycleavePCR in 136nasopharyngeal suction specimens. The CycleavePCR results were consistent with those of routine PCR and sequencing. There was no signal production from other bacterial species. Sensitivity and specificity were 100%. The detection limit of the CycleavePCR was 10 plasmid copies per reaction. Experiment can be done within 1.5 h. Conclusion A novel method is developed to detect erythromycin-resistant strains harboring A2063G and A2064G transition mutation in the 23s rRNA gene using CycleavePCR.