Objective To explore the establishment of peptide mapping database of Candida albicans ,laying the foundation for rapid diagnosis of Candida albicans infection .Methods 96 Candida albicans were collected clinically ,and its DNA was extracted . Polymerase chain reaction(PCR) was used to amplify the ITS1-5 .8S-ITS2 gene fragments and restriction endonucleases were a-dopted to identify them .Surface enhanced laser desorption ionization-time of flight-mass spectrometry(SELDI-TOF-MS) instrument was applied to detect the Candida albicans peptide mapping ,and Ciphergen ProteinChip software was used to collect data automati-cally .The established peptide mapping database was verified by confirmed Candida .Results According to restriction fragment length polymorphism analysis ,96 strains were confirmed as Candida albicans .15 peptide peaks were captured by SELDI-TOF-MS chips .Five peptide peaks of them with stable expression were screened out ,and the similarity analysis software was used to estab-lish peptide mapping database of Candida albicans .More than 95% of similarity was found between peptide mapping of Candida albicans and established database ,while less than 50% was found between peptide mapping of other Candida species and database . Conclusion The establishment of peptide mapping database of Candida albicans provides a theoretical basis for the rapid diagnosis of Candida albicans infection .

ObjectiveTo sum up the experience in the treatment of acute arterial ischemia in the extremities. Methods From 1980 to 2001,148 patients with acute arterial ischemia in the extremities were treated by multiple-means such as: embolectomy, interventional treatment, thrombolytic and antiagglutinatives. Results The cure rate in patients treated within 12 hours was 95.5%,mortality was 4.5%,while the cure rate, alleviative rate, amputation rate and mortality in patients treated 12～24 hours after onset were 64.8%,17.6%,9.9%,7.7%,respectively and that were 20%,34.3%,25.7%,20% respectively when treatment started 24 hours after the onset. The cure rate in 19 patients treated by nonoperative means was 10.5%, alleviative rate was 73.3%, amputation rate was 15.8%. Conclusion Patients with acute arterial ischemia suffer a high mortality. Mortality and disability rate can be reduced by early diagnosis, appropriate treatment and effective management for the systemic diseases.

Objective To establish protein fingerprints of common bacteria in clinics and to lay a foundation for rapid identification of bacteria.Methods Strains of Escherichia coli,Klebsiella pneumoniae,Pseudomonas aeruginosa and Staphylococcus aureus were detected by surface enhanced laser desorption and ionization time-of-flight mass spectrometry (SELDI-TOF MS).Stable expression protein peaks were screened and the data was input into the self-constructed Fingerwave software for identification of target bacteria by protein fingerprint comparison.Two hundred and fifty-six clinical isolates,including E.coli,K.pneumoniae,P.aeruginosa and S.aureus were detected and the data was compared with constructed database to evaluate its diagnostic value.Results The protein fingerprints including four common bacteia was used to identify the target bacteria with identification rate of 93.1％ (54/58) for E.coli,87.2％ (75/86) for K.pneumoniae,96.2％ (60/63) for P.aeruginosa and 96.2％ (51/53) for S.aureus,respectively.Conclusion Common bacteria can be rapidly identified by using the protein fingerprint comparison,which provides a powerful tool for bacterial identification.

Objective To investigate the mechanism of one carbapenems resistant Raoultella planticola( R.planticola) isolate.Methods This is an experimental study.R.planticola was isolated from a patient′s drainage fluid from orthopedic department in November 2010 in Sichuan Provincial People′s Hospital.Minimum inhibitory concentration of R.planticola to 13 antibiotics was determined by using the agar dilution method.Modified Hodge test was used to detect carbapenemase .EDTA synergistic test was performed to research metallo-beta-lactamase.The genes coded the β-lactamase were amplified by polymerase chain reaction ( PCR ) , including class A carbapenemase ( KPC ) , class B carbapenemases (NDM, IMP, VIM, SIM), extended spectrum beta-lactamases[ESBL(CTX, TEM, SHV)], and AmpCβ-lactamases ( FOX, EBC, ACC, DHA, CIT, MOX).Results The susceptibility test showed that R.planticola was resistant to 9 antibiotics.MIC value of meropenem for R.planticola was up to 32 mg/L.R.planticola kept intermediary to imipenem , whereas it was susceptible to cefepime , amikacin and polymyxin B.Modified Hodge test and EDTA synergistic test were positive in R.planticola.Class B carbapenemase (IMP) gene and two extended spectrum β-lactamases(CTX, SHV) genes were positive by PCR.The genes were conformed as IMP-4, CTX-M3 and SHV-12 by sequencing and compared with GenBank.Other resistant genes were negative.Conclusion IMP-4 was identified in R.planticola, the combined produce IMP-4 and ESBLs might be the main mechanism of R.planticola resistant to carbapenems.

Objective To construct mutant strains of methicillin resistant Staphylococcus epidermi-dis (MRSE) with psm-mec gene deletion and to investigate the function of psm-mec gene.Methods The drug sensitivity test and DNA sequence analysis were performed to screen out the tetracycline and chloram -phenicol sensitive clinical strains of MRSE , whose upstream and downstream sequences of psm-mec gene were identical to those of the Staphylococcus epidermidis reference strain RP62A.The recombinant plasmid pBT2-Δpsm-mec was constructed by using the fusion PCR and a temperature sensitive shuttle plasmid .After being identified , the plasmid was transformed into the Staphylococcus aureus RN4220 strain by electropora-tion, and then transformed into the selected clinical isolates of MRSE .The mutant strains of MRSE with psm-mec deletion were screened out and identified after homologous recombination .The differences in biofilm formation between the mutant and wild-type strains were analyzed for further elucidation the relationships be-tween the psm-mec gene and biofilm formation in MRSE strains .Results Three clinical MRSE isolates for the construction of mutant strains with psm-mec gene deletion were screened out and identified by using drug sensitivity test and sequence alignment analysis .The mutants constructed via homogenous recombination were screened out and identified .Compared with the corresponding wild-type strains, the three mutants with psm-mec gene deletion showed significantly decreased ability of biofilm formation , demonstrating that the psm-mec genes strains induced the biofilm formation of MRSE .Conclusion The Δpsm-mec mutant strains were successfully constructed .The psm-mec gene played an important role in the biofilm formation of Staphy-lococcus epidermdis.

Objective To investigate the genetic location of SCCmec-associated psm-mec in Staphylococcus hominis isolated from blood culture,and to lay a foundation for further functional studies of psm-mec in Staphylococcus hominis.Methods 25 strains of Staphylococcus hominis isolated from positive blood culture were collected.mecA and psm-mec gene were amplified by PCR,and the SCCmec types were determined by the results of multiplex PCR assay.For analyzing the genetic location characteristic of psm-mec in SCCmec,three pair special PCR primers were used to measure mecR1/psm-mec,psm-mec/xylR and fudoh respectively.Results There were 21 strains of methicillin-resistant Staphylococcus hominis and 4 strains of methicillin-sensitive Staphylococcus hominis. The positive rate of psm-mec gene in methicillin-resistant Staphylococcus hominis was 47.6%,and no psm-mec gene was found in methicillin-sensitive Staphylococcus hominis.Among psm-mec positive strains,2 strains belonged to SCCmecⅢ,5 strains belonged to SCCmecⅢ-like,and 3 strains belonged to new SCCmec types.All of the 10 psm-mec positive strains were mecR1/psm-mec,psm-mec/xylR and fudoh gene positive.Conclusion SCCmec-associated psm-mec extensively exists in methicillin-resistant Staphylococ-cus hominis isolated from positive blood culture,which distributes mainly in typical SCCmecⅢ,SCCmecⅢ-like and new SCCmec types and locates between mecR1 and xylR gene.

Objective To investigate the SCCmec types of clinically isolated methicillin‐resistant Staphylococcus epidermidis (M RSE) .Methods Eighty‐four strains of clinically isolated Staphylococcus epidermidis identified by the fully automatic microbio‐logical identification system were collected and performed the MRSE identification by PCR for amplifying esp and mecA genes and SCCmec typing .Its distribution characteristics were analyzed .Results Esp gene was amplified in 84 strains and the detection rate of mecA was 76 .19% (64/84) ,in which the MRSE detection rates in blood ,sputum ,urine and wound secretion were 76 .8% , 68 .8% ,100% and 71 .4% respectively .The multiple PCR amplification displayed that among 64 strains of MRSE ,19 strains were SCCmec simple type ,in which 19 strains were SCCmec type Ⅰ and 3 strains were SCCmec type Ⅲ ;42 strains were SCCmec mixed type ,in which 2 strains were SCCmec mixed type Ⅰ and Ⅱ ,14 strains were SCCmec mixed type Ⅰ and Ⅲ ,12 strains were SCCmec mixed type Ⅰ ,Ⅱ and Ⅲ ,5 strains were SCCmec mixed type Ⅱ and Ⅲ ,a strains were and SCCmec mixed type Ⅲ and Ⅳ .Conclu‐sion The SCCmec type in clinically isolated MRSE shows obvious diversity and its majority is SCCmec mixed type .

Objective To observe the histological effect of acellular dermal matrix(ADM)on insufficiency in guide keratinized tissue regeneration. Methods 6 cases of single anterior tooth implantation in the hospital in 2016 were included in the study. 3 cases were treated with immediate tooth extraction and implantation. Bone substitution materials were grafted in the space between the tooth extraction socket and the implants. The keratin-ized tissue dehiscence was covered by double layers of acellular dermal matrix membrane(Heal-All?,ZH-BIO, China),which was fixed to the adjacent soft tissue by suturing. Another 3 cases were routinely treated with delayed implantation of single anterior tooth. All the cases were subject to harvesting of the cover soft tissues of implants with a punch 4 months later. The new grown soft tissues were histologically observed. Results All cases were sur-vived. The new grown keratinized tissues were observed. Conclusion Acellular dermal matrix can guide the aug-mentation of keratinized tissues.

Objective To discuss the application value of modified Hodge test(MHT) for screening carbapenemase-producing Enterobacteriaceae.Methods The 24 Enterobacteriaceae reduced susceptibility to carbapenems were detected by MHT.At the same time,polymerase chain reaction(PCR) was used to detect carbapenemase genes of KPC,NDM,IMP,SIM and VIM.PCR products were sequenced and the results were compared with the sequences of Gen Bank database.Comprehensive analysis the application value of MHT and PCR to detect carbapenemase.Results Among these 24 strains,13 stains appeared to produce carbapenemase by MHT,5 positive strains were found to carry carbapenemase genes by PCR.By comparing with the sequences of Gen Bank database 1 strain were confirmed to KPC-2 and 4 strains were confirmed to IMP-4.We found that 4 strains of Enterobacteriaceae,detected carbapenemase by MHT and PCR at the same time.9 strains of MHT were positive,but we couldn′t detect the carbapenemase genes.1 strain of MHT was negative,but carbapenemase gene was found in the strain.Conclusion The value of MHT to screen carbapenemase-producing Enterobacteriaceae is necessary to further study.

Objective To investigate the perioperative risk factors associated with prognoses of patients with acute ischemic stroke accepted mechanical thrombectomy under general anesthesia.Methods The clinical data of 108 patients with acute ischemic stroke,admitted to and accepted mechanical thrombectomy under general anesthesia in our hospital from January l,2016 to October 31,2018,were collected.According to modified Rankin scale (mRS) scores 90 d after surgery,patients were divided into good prognosis group (mRS scores ≤2) and poor prognosis group (mRS scores ≥3).Univariate analysis was used to compare the general data (age,gender,body mass index,and underlying diseases) and perioperative conditions (immediate heart rate,systolic and diastolic blood pressures immediately after admission,operative time,and anesthesia time) between the two groups of patients.Multivariate Logistic regression analysis was used to identify the perioperative risk factors influencing the prognoses of patients with acute ischemic stroke accepted mechanical thrombectomy.Results Among the 108 patients,65 had good prognosis and 43 had poor prognosis.Univariate analysis showed that there was no significant difference in general data between the two groups (P＞0.05),but there were significant differences in heart rate immediately after admission,National Institutes of Health Stroke Scale (NIHSS) scores immediately after admission and 3 d after operation,maximum hemoglobin and blood glucose values from immediately after admission to 3rd d of operation,and thrombolysis in myocardial infarction (TIMI) blood flow classification (P＜0.05).Multivariate Logistic regression analysis showed that heartrate immediately after admission (OR=1.035,95％CI:1.002-1.067,P=0.037) and NIHSS scores 3 d after operation (OR=1.153,95％CI:1.016-1.272,P=0.030) were the perioperative risk factors influencing the prognoses of patients with acute ischemic stroke accepted mechanical thrombectomy.Conclusion For patients with acute ischemic stroke who have rapid heart rate immediately after admission and high NIHSS scores 3 d after mechanical thrombectomy,possibility of poor prognosis should be noticed.