OBJECTIVE To investigate the resistant mechanisms of Candida albicans to azoles at molecular level.METHODS NCCLS M-27 protocols were used to test the in vitro susceptibilities of 102 C.albicans strains isolated from the patients with recurrent vulvovaginal candidiasis(RVVC) against fluconazole(FLC) and itraconazole(ITC) to screen the FLC-and ITC-resistant C.albicans isolates;six pairs of primers,A1-A2,B1-B2,C1-C2,D1-D2,E1-E2 and F1-F2 were respectively to amplify gene CYP51 of 4 strains with FLC-and ITC-resistance.The PCR products were sequenced and analyzed to identify the mutation sites by compared with the sequence of gene CYP51 of referenced C.albicans strain in NCBI site of Internet.RESULTS The analysis of full length sequence of CYP51 from 4 FLC-and ITC-resistant strains showed that from total 32 mutation sites there were 4 significant site mutations,where the mutation of GAT to GAC at 116 caused the substitution of D by E(E266D in two strains);GCC to GGT at 117 caused the substitution of A by G(A117G in 1 strain);GAA to GAC at 266 caused the substitution of E by D(E266D in 2 strains);and GTT to ATT at 488 caused the substitution of I by V(V488I in 1 strain).The site mutations of 266 and 488 were tested in 1 strain of 4 strains.CONCLUSIONS The CYP51 total gene of 4 strains has been checked out.Of FLC and ITC-resistant C.albicans alignment in this time,find out 4 significant bp mutations.Causing its amino acide change,among them,A117G has not be interrelated report still now.The details of mechanism need to be further studied.

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 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 In order to understand which kind of function genes play an important role for es-timating wound age, the variation of difference genes’ mRNA expression were compared after injury. Methods T he mRNA expression levels of seven candidate genes (ICAM-1, NF-κB, MX2, MT1, MT2, sTnI, and Cox6c) were analyzed in contused rat skeletal muscle at different time points using real-time fluorescent quantitative PC R (R T-qPC R ). T he rawC t values were normalized relative to that of RPL32 mRNA , and converted to standard C t values. A t each time point after injury, the standard deviations (SD ) of the standard C t values were calculated by SPSS. Results T he expression trends of the seven genes were all found to be related to wound age, but there were lower variation coefficients and greater reliability of sTnI and Cox6c when compared with other genes. Conclusion T he genes encoding struc-tural proteins or proteins that performbasic functions can be suitable for wound age estimation.

Objective T o investigate the relationship betw een the expression of secreted frizzled-related protein 5 (SFR P5) m RNA and the tim e interval after skeletal m uscle injury in rats by real-tim e PC R . Methods A total of ninety SD rats w ere random ly divided into the contusion groups at different tim es including 4 h, 8 h, 12 h, 16 h, 20 h, 24 h, 28 h, 32 h, 36 h, 40 h, 44 h, 48 h after contusion, incision groups at different tim es including 4h and 8h after incision and the control group. T he sam ples w ere taken from the contused zone at different tim e points. T he total RNA w as isolated from the sam ples and re-versely transcribed to analyze the expression levels of SFRP5 m RNA . Results C om pared to the control group, the expression of SFRP5 m RNA in contusion groups w ere dow n-regulated w ithin 48 h after con-tusion and reached the low est level at 20 h, and the expression of SFRP5 m RNA gradually increased from 20 h to 48 h after contusion. T he expression of SFRP5 m RNA in the incised groups w ere signifi-cantly low er than that of the contusion groups at 4 h after injury. A t the tim e of 8 h, the expression levels betw een the contusion and incision groups show ed no statistically significant difference. Conclusion It is suggested that SFRP5 m RNA analysis m ay show regular expression and can be a m arker for estim ation of skeletal m uscle injury age.

Objective To investigate the effect of B TX -A block by insulative needle to release the spasticity in cerebral palsy. MethodsFrom June 2000 to November 2002, 47 children wit h spastic cerebral palsy aged 2 to 15 years old, averaged( 6.32?2.52) years o ld were divided into an experiment group and a control group. The 33 children in the experiment group were blocked with BTX-A by insulative needles. The other 14 children in the control group were blocked with BTX-A by use of the convent ional syringe. ResultsThe spasticity in the experim ent group reduced to a larger extend than that in the control group, and the th erapeutic effect lasted longer in the experiment group. Conclusi onUsing insulative needle can improve the effect of BTX-A block.

Objective To compare the advantages and disadvantages of botulinum toxin A (BTX-A) and phenol block in the treatment of spasticity in children with cerebral palsy. Methods Four hundred and twenty children with spastic cerebral palsy were divided into an experimental group (375 cases) and a control group (45 cases).The children were aged from 1 to 22 years ( average age 6 years).The children in the experimental group were treated with BTX-A block at a dosage of 55 to 350 IU (average 130.5 IU).The children in the control group were treated with a 5％ phenol solution block at a dosage of 0.5 to 4.6 ml ( average 2.2 ml).Children of both groups were given systematic functional rehabilitation training. All the children were evaluated with a physician rating scale (PRS) and the modified Ashworth scale (MAS) before and after the blocking.Effectiveness rates,effectiveness durations and side effects rates were calculated. Results Before treatment there was no significant difference in terms of motor disorder or spasticity between the 2 groups.After treatment,spasticity had been significantly reduced in both groups.The effectiveness rate was 98.4％ in the experimental group and 95.6％ in the control group,a difference which was not significant.The average effectiveness duration was ( 24.9 ± 5.76 ) weeks in the experimental group and ( 69.2 ± 13.76) weeks in the control group,significantly longer.The side effects rate was 5.33 ％ in the experimental group and 15.56％ in the control group,also a significant difference. Conclusion BTX-A could be more widely used because of its safety and credibility.

Objective To characterize clinical feature, frequency of isolation and antimicrobial susceptibility of pathogens isolated from patients with nosocomial bloodstream infections in Huashan Hospital, Fudan University from 1995 to 2004. Methods The clinical data of all patients who were diagnosed with nosocomial bloodstream infections based on national diagnostic criteria of nosocomial bloodstream infections were retrospectively analyzed. The pathogens were routinely isolated and identified. Susceptibilities against antimicrobial agents were determined by Kirby-Bauer methods and analyzed by WHONET 5.0 software. Results During the 10-year study period, a total of 395 patients were diagnosed with nosocomid bloodstream infection with 435 strains isolated from blood specimen.Gram positive bacteria, Gram negative bacilli and fungi, accounted for 47.4%, 45.1 % and 7.6%,respectively. Coagulase-negative Staphylococci (21.4%), S. aures (17.9%), E.coli (13.6%), K. pneumoniae (10.8%), Candidaspp (7.4%), Enterococci (6.0%), Pseudomonasspp (6.0%) and Acinetobacter spp (3.7%) were frequently identified isolates. S. aures and coagulase-negative Staphylococci resistant to methicillin were 62.8% and 87.1%, respectively. The susceptibilities of cefotaxime and ceftazidime against E. coli and K. preumonine were 46%-78% and 27.7%-40.4%, respectively. Conclusions The Gram positive cocci are slightly more prevalent than Gram negative bacilli in nosocomial bloodstream infections and resistance to the first line antibiotics is common among all pathogens isolated. Candida spp is the fifth leading cause of nosocomial bloodstream infections.

Objective To investigate the optimal dose of botulinum toxic A(BTX-A)for releasing spasticity of cerebral palsy(CP)and related factors.Methods 39 children with CP was treated by BTX-A injected in local muscles to release the spasticity.Results After treatment,23 children had good curative effect.In them,15 cases were male,8 cases were female,the mean age was 60.7±26.9 months(range 33 to 145 months).There was no correlation between the optimal unit dose of BTX-A and those including sex,age,body weight,diagnosis type,degree of Gross Motor Function Classification System(GMFCS),etiological factor and the injections.There was a linear positive correlation between the optimal unit dose of BTX-A and the score of Modified Ashworth Scale(MAS).Conclusion BTX-A can release the spasticity of CP children,the injecting dose is correlated with MAS score.

Objective To investigate whether the curative effect of Botulinum Toxic A(BTX-A) block increased and prolonged when the dose calculated according to certain formula.Methods 15 children with cerebral palsy(CP) were appointed as the trial group and the dose of BTX-A was calculated according to the designed formula.The other 58 CP children were appointed as the control group and the BTX-A dose was determined by routine method.Results After treatment,the motor function of the children in trial group was superior to that in the control group(P<0.05) and the duration of therapeutic effect of the trial group was significantly longer than the control group(P<0.001).Conclusion BTX-A dose calculated according to the formula can increase effect and prolong the duration.