Objective To establish a method for detecting Aspergillus spp.by Loop-mediated isothermal amplification.Methods Aspergillus spp.specific primers were designed from the relative conservation region of the published sequence of 28S rRNA genes of A.fumigatus (GenBank accession number AY660917),A.terreus (GenBank accession number AF454183),A.flavus (GenBank accession number AF454158),and A.niger (GenBank accession number AF454169).Genomic DNA were extracted from Aspergillus standard strains,clinical control strains and clinical samples,and amplified by LAMP.The amplified results could be read with the naked eye by the coloring effect of fluorescent nucleotide dye without the DNA electrophoresis.Approximately 103 each Aspergillus conidia suspension was added to the serum from healthy volunteers,and detected these simulative clinical samples bv LAMP assay.Results The LAMP and PCR assays obtained positive results for all four Aspergillus species and 8 simulative clinical samples (double samples for each Aspergillus species) including 103 conidia,but negative in the remaining 15 non-Aspergillus species,human total blood genomic DNA,30 clinical serum samples infected with non-Aspergillus and 10 healthy volunteers.The LAMP assay had a minimum detection of 0.05-0.5pg,by means of detect different levels of 500,50,5,0.5,0.05 pg template in each reaction tube.Conclusion The results confirm that LAMP is a simple,rapid,sensitive and specific method,and can be used for detection of Aspergillus strains in clinical and environmental specimens.

OBJECTIVE To study the existence status of class Ⅰ integron and transposable element of multi-resistant Acinetobactor baumannii isolated form ICU of Yinzhou People′s Hospital in Ningbo. METHODS The samples of 20 A.baumannii isolates were collected from Oct 2007 to Jul 2008.The susceptibility to 32 antibiotics of the isolates was measured.The genetic markers of integron qacE△1-sul1 and transposable element tnpU were analyzed by polymerase chain reaction(PCR).The PCR products of tnpU or qacE△1-sul1 were sequenced for determination. RESULTS In the 20 ABA isolates,the positive rate of class Ⅰ integron qacE△1-sull was 75%,and the positive rate of transposable element tnpU was 55.0%. CONCLUSIONS The positive rate of the integron qacE△1-sul1 and transposable element tnpU for multi-resistant A.baumannii is high in Yinzhou People′s Hospital in Ningbo.It should be reevaluated the preventative role of chlorhexidine for operation.

OBJECTIVE To investigate the antibiotics resistance of multi-resistant Acinetobactor baumannii(ABA) and genotypes of beta-lactamases in ICU.METHODS The samples of 20 A.baumannii isolates were collected from Oct 2007 to Jul 2008 from patients in ICU.To determine the sensitivity to the 32 kinds of antibacterials,K-B method was used and the detection of ESBLs and AmpC beta-lactamases was performed by three dimensional test and 21 types of beta-lactamases genes were analyzed by polymerase chain reaction(PCR).RESULTS Among the 20 ABA isolates,all carried TEM beta-lactamases gens(100%),10 carried OXA-23 beta-lactamases gens(50%) and 15 strains carried ADC beta-lactamases gene(75%),50% strains produced TEM,OXA-23 and ADC beta-lactamases simultaneously.Through determining sequence of one PCR product from TEM,OX23 and ADC respectively,we found TEM-116,OXA-73 and ADC-25 type beta-lactamases genes.CONCLUSIONS The antibiotics resistance of ABA is very serious.TEM,OXA-23 and ADC exist in multi-resistant A.baumannii widely.It should be the main causes as high rate of drug-resistantce to beta-lactamantibiotics.

OBJECTIVE To investigate the antibiotics resistance of Escherichia coli(ECO) and distrubtion of aminoglycoside-modifying enzymes and 16S RNA methylase genes in ICU.METHODS The samples of 20 ECO isolates were collected from Dec 2007 to Jun 2008 of patients in ICU.To determine the sensitivity to the 30 antibacterials K-B method was used and aminoglycoside-modifying enzymes and 16S RNA methylase genes were analyzed by polymerase chain reaction(PCR).RESULTS In among the 20 ECO isolates,8 strains carried aac(3)-Ⅱ(40%),3 carried aac(6′)-Ⅰb(15%) and ant(3″)-Ⅰ(15%),1 be found aph(3′)-Ⅰ(10%)and 13 be found aadA4/5 aminoglycoside-modifying enzymes genes,no strain carried 16S RNA methylase genes.CONCLUSIONS aac(3)-Ⅱ、aac(6′)-Ⅰb,ant(3″)-Ⅰ,aph(3′)-Ⅰ and aadA4/5 aminoglycoside-modifying enzymes genes exist in ECO widely,they should be the main cause inducing the high rate of drug-resistance to aminoglycosides.

OBJECTIVE To study the mechanisms of chromosome and plasmid-mediated quinolones resistance in Escherichia coli(ECO).METHODS Clinical isolates of ECO were collected from clinical specimens at ICU of Yinzhou People′s Hospital from Dec 2007 to Jun 2008.To detect the susceptibility to 30 types of antibiotics K-B disk diffusion method was used.The susceptibicity to ciprofloxacin and levofloxacin were detected by agar dilution testing.Then six type genes gyrA,qnrA,qnrB,qnrS,aac(6′)-Ⅰb-Cr,and qepA were investigated by PCR.In the meantime,the PCR products were sequenced.RESULTS The alterations in gyrA were found in all 20 tested strains.Two new subtypes were found in ECO 13 and ECO 15.Three strains were found acc(6′)-Ⅰb-Cr and ECO 2 was detected qurA gene.CONCLUSIONS The mutations in gyrA play a dominant role in the resistance to quinolones in ECO.Both aac(6′)-Ⅰb-Cr and qepA may responsible for quinolones resistance in ECO too.

OBJECTIVE To investigate mycoplasma infection in ICU patients.METHODS Sixty-five samples from blood,respiratory tract and genitourinary tract of patients were collected respectively from Oct 2007 to July 2008 in ICU.Mycoplasma pneumoniae(Mp),Urealasma urealytium(Uu) M.fermentans(Mf) and M.penetrans(Mpe) were cultivated by modified mycoplasma fluid and solid medium.Mf and Mpe positive isolates were verified by nested polymerase chain reaction(rPCR),Mp and Uu were confirmed by fluorescent quantitative PCR.RESULTS It was found that the positive detection rate for Mp was 12.3%(8/65)in blood and 35.4%(23/65) in respiratory tract excreta and for Mu 1.5%(1/65) and 26.2%(17/65) in blood or Genitourinary tract,respectively.Mpe and Mf did not detected.CONCLUSIONS The state of mycoplasma infection is very severe,and often accompanies bacterial infection.It is necessary to consider mycoplasma when chose antibiotics.

Objective To investigate the distribution of virulence-related genes in multidrug resistant Escherichia coli.Methods Seven virulence genes papA,cnf1,cnf2,cfaB,ipaB,hofQ and ompT were detected by PCR in 20 strains of multidrug resistant Escherichia coli clinically isolated,and the positive genes were further searched in 31 strains of Escherichia coli in BioCyc database whose genomies had been fully sequenced.Results Virulence genes hofQ and ompT were detected in 20 strains of Escherichia coli with a positive rate of 95.0％ (19/20) and 55.0％ ( 11/20),respectively.Among 31 strains of Escherichia coli in BioCyc,21 (67.7％) were positive for hofQ gene and 15 (48.4％) were positive for ompT gene.Conclusion hofQ and ompT genes are prevalent in multidrug resistant Escherichia coli.

OBJECTIVE To investigate the antibiotic resistance of multi-resistant Acinetobacter baumannii(ABA) and distribution of aminoglycoside-modifying enzymes and 16S rRNA methylase genes in ICU in Yinzhou People′s Hospital in Ningbo. METHODS The samples of 20 ABA isolates were collected from Oct 2007 to Jul 2008 in ICU.K-B method was used to determine the sensitivity to 32 antibacterials and the aminoglycoside-modifying enzymes and 16S rRNA methylase genes were analyzed by polymerase chain reaction(PCR). RESULTS From 20 ABA isolates,8 strains carried aminoglycoside-modifying enzymes genes aac(3)-Ⅰ,aac(3)-Ⅱ,aac(6′)-Ⅰb,and ant(3″)-Ⅰ,their positive rate was 10%,15%,30% and 25%,respectively;no strain carried 16S rRNA methylase genes. CONCLUSIONS The antibiotics resistance of A.baumannii is very serious in Yinzhou People′s Hospital in Ningbo.Aminoglycoside-modifying enzymes genes aac(3)-Ⅰ,aac(3)-Ⅱ,aac(6′)-Ⅰb and ant(3″)-Ⅰ exist in multi-resistant A.baumannii widely.They would be the main causes of high drug-resistantce to aminoglycosides.

Objective To evaluate the impact of postoperative adjuvant chemotherapy on prognosis of patients with advanced rectal carcinoma using a meta-analysis.Methods We searched PubMed to identify literature comparing observation with adjuvant chemotherapy after neoadjuvant chemoradiotherapy and surgery for patients with advanced rectal carcinoma.Data were analysed using Revman 5.0 statistical software.Results Nine trials were included consisting of 6 212 patients:3 421 patients received adjuvant chemotherapy and 2 791 patients did not.The age ranged from 55.6 to 68 years.Adjuvant chemotherapysignificantly affects overall and disease-free survival (P =0.002 and P =0.000 5 respectively) of patientswho had received neoadjuvant chemoradiotherapy.The subgroup analysis which originate from insufficient data reveales ypT0-2 patients and ypT3-4 patients can't benefit from adjuvant chemotherapy.Conclusion Postoperative adjuvant chemotherapy can improve the survival of advanced rectal carcinoma patients after neoadjuvant chemoradiotherapy,but can't increase the survival of pathology complete respone and yPN + patients.

OBJECTIVETo detect potential mutations of EDA gene for a Chinese family affected with X-linked hypohidrotic ectodermal dysplasia (XLHED).

METHODSGenomic DNA was extracted from peripheral blood of the proband, his relatives and 50 non-related healthy controls. Exonic sequences of the EDA gene were subjected to polymerase chain reaction amplification and direct sequencing.

RESULTSA c.467G> A mutation (R156H) was detected in exon 3 of the EDA gene in the proband, his mother, 2 uncles, and 1 aunt. The same mutation was not detected in the 50 non-related healthy controls.

CONCLUSIONA c.467G>A mutation of the EDA gene probably underlies the disease in the family.

Base Sequence ; Child ; Ectodermal Dysplasia 1, Anhidrotic ; diagnosis ; genetics ; Ectodysplasins ; genetics ; Exons ; Female ; Genotype ; Humans ; Male ; Mutation ; Pedigree