Analysis of drug resistance genes and molecular typing of carbapenem-resistant Klebsiella variicola strains / 中华微生物学和免疫学杂志

Objective To investigate the prevalence and drug resistance of clinical Klebsiella vari-icola ( K. variicola ) isolates and to illuminate the mechanism of drug resistance in carbapenem-resistant strains. Methods Clinical K. variicola isolates were identified with matrix-assisted laser desorption/ioniza-tion time-of-flight mass spectrometry ( MALDI-TOF MS ) . The antimicrobial susceptibility profile of these strains was determined using broth microdilution. Resistance genes carried by carbapenem-resistant K. vari-icola strains were detected by PCR with specific primers. Multilocus sequence typing ( MLST) was used for molecular typing. A pan-drug resistant strain which was isolated from cerebrospinal fluid sample was ana-lyzed with whole genome sequencing ( WGS) . Results Twenty-six isolates were identified as K. variicola by MALDI-TOF MS. Results of the antimicrobial susceptibility test showed that there were 15. 4％ (4/26) re-sistant to carbapenem and 11. 5％ (3/26) unsusceptible to tigecycline. These strains were highly suscepti-ble to amikacin and gentamicin, which accounted for 96. 2％ (25/26). As for the third-and fourth-genera-tion cephalosporins, the resistance rate was 23. 1％ (6/26). All of the four carbapenem-resistant isolates carried the resistance genes of blaIMP-4 , qnrA/B and blaTEM , and one of them was also positive for blaNDM-1 gene. The fosfomycin resistance gene, fosA, was detected in three of them. Molecular typing analysis indica-ted these isolates belonged to two sequence types ( ST) of ST357 ( three strains) and ST1737 ( one strain) . Two plasmids were obtained from the pan-drug resistant strain by WGS, including IncFⅡ/FIB( k) type plas-mid (160 kb) that was highly homologous to LMG 23571 plasmid (GenBank: CP013986. 1) and IncHⅠ1B/FIB type plasmid (260 kb) sharing high homology with pIMP4 LL34 (GenBank: CP025964. 1). Be-sides the resistance genes mentioned above, the two plasmids also carried a variety of other genes that media-ted the resistance to aminoglycosides (strB, strA, armA, aac3-Ⅱd, aadA2), macrolides (msrE, mphE), chloramphenicol (catA2), sulfonamides (sulⅠ) tigecycline (tetA variant) and trimethoprim (dfrA16). However, no virulence genes were detected. Conclusions In general, the resistance profile of K. variicola was similar to that of Klebsiella pneumoniae, but the differences were that carbapenem-resistant K. variicola strains mainly belonged to ST357 and the leading causes of resistance were carrying the genes encoding IMP-4 and NDM-1 metalβ-lactamases. WGC analysis revealed that the pan-drug resistant K. variicola strain carried multiple drug resistance genes without virulence determinants, which might be resulted from the evo-lution of drug resistance.

Twenty-eight cases of patients with acute closed rupture of the achilles tendon of five years follow-up study / 中华显微外科杂志

Objective To compare the curative effect of open and minimally invasive treatment of acute closed achilles tendon rupture with 5 years followed-up study.Methods From September,2010 to January,2012,28 patients with acute closed Achilles tendon rupture in our hospital were followed up for 5 years.There were 21 males and 7 females.Minimally invasive percutaneous suture in 11 cases;open suture in the treatment of 17 cases.The patients were followed up at 6 months,1 year,2 years,3 years,4 years,and 5 years after AOFAS and ATRS score.Results The follow-up time ranged from 60 to 72 months.All incisions healed by first intention and no incision related complications occurred.2 groups of patients with ATRS score at 6 months after operation:the open group was 81.23±3.99,minimally invasive percutaneous group of 88.27±4.27,the difference between the two groups was statistically significantly.After 1 year,there was no significant difference in the scores between the two groups.Two groups of patients with AOFAS score at 6 months after operation:the open group was 69.00±6.23,minimally invasive percutaneous group of 79.27±4.83,the difference between the two groups was statistically significantly.At one year after operation,the open group was 85.53±3.38,and the minimally invasive group was more than 89.90±3.38.The difference between the two groups was statistically significantly.After 2 years,there was no significant difference in the scores between the two groups.Conclusion There is no significant difference between the 2 years after surgery in the treatment of acute closed Achilles tendon rupture or open surger.

Effects of early physiotherapy in combination with atorvastatin on the levels of serum brain-derived neurotrophic factor and neurological function in patients with acute ischemic stroke:a randomized controlled study / 国际脑血管病杂志

Objective To investigate the effects of early physiotherapy in combination with atorvastatin on the levels of serum brain-derived neurotrophic factor (BDNF) and neurological function in patients with acute ischemic stroke.Methods Fifty patients with acute ischemic stroke were randomly divided into either an atorvastatin group (monotherapy group,n =25) or a early physiotherapy + atorvastatin group (combination treatment group,n =25).All patients received the prescribed drugs according to the diagnosis and treatment guidelines for ischemic stroke.The monotherapy group added atorvastatin calcium (20 mg,1 tablet every night orally).On the basis of the monotherapy group,the combination treatment group also conducted early physical therapy.At 2 and 6 weeks before and after treatment,a double-antboody sandwich enzyme-linked immunosorbent assay was used to detect the serum BDNF levels.The National Institutes of Health Stroke Scale (NIHSS) was used to evaluate the degree of neurological deficit.Barthel index (BI) was used to evaluate the activities of daily living.The modified Rankin scale (mRS) was used to assess the degree of disability.Results There was no significant difference in demographics and baseline data between the monotherapy group and the combination treatment group.The scores of NIHSS,BI,and mRS in both groups after treatment were significantly better than those before treatment (all P ＜ 0.001).There were no difference in the scores of NIHSS,BI and mRS at 2 weeks before and after treatment,but at 6 weeks after treatment,the scores of NIHSS (2.40 ± 1.38 vs.3.36 ± 1.73; P =0.035) and mRS (1.40 ± 0.87 vs.1.96 ±0.94; P =0.047) of the combination treatment group were significantly lower than those of the monotherapy group,and the BI scores (92.60 ±7.50 vs.85.20 ± 11.68; P=0.011) were significantly higher than those of the monotherapy group.After treatment,the serum BDNF levels were increased significantly in both groups.There were significant differences among all the time points (all P＜0.001).At 2 weeks after treatment,the serum BDNF levels (3.07 ±0.93 ng/ml vs.2.45 ±0.76 ng/ml; t =2.559,P =0.014) and at 6 weeks after treatment,those (2.90 ± 0.93 ng/ml vs.2.31 ± 0.77 ng/ml; t =2.433,P =0.019) in the combination treatment group were significantly higher than those in the monotherapy group.Spearman correlation analysis showed that the serum BDNF levels were significantly negatively correlated with the scores of NIHSS (r =-0.738,P ＜ 0.001) and mRS (r =-0.654,P ＜ 0.001),but they were significantly positively correlated with the BI scores (r =0.716,P ＜ 0.001).No serious adverse reaction occurred in both groups.Conclusions Early physiotherapy in combination with atorvastatin for the treatment of acute ischemic stroke can more effectively promote the recovery of neurological function,and its mechanism may be associated with the increased serum BDNF levels.

Brain-derived neurotrophic factor and cerebral ischemia / 国际脑血管病杂志

Brain-derived neurotrophic factor (BDNF) is one of the most prevalent growth factors in the central nervous system (CNS).In the development and maturation processes of the nervous system,BDNF plays an important role in maintaining neuronal function,promoting neuronal regeneration after injury,and preventing neuronal degeneration,etc.At present,many researchers are being dedicated to the research of BDNF for treatment of brain ischemia and have achieved some progress.This article reviews the molecular biological characteristics and biological function of BDNF,roles and mechanisms in cerebral ischemia,and the possibility as an intervention target of cerebral ischemia.

Species identification of Acinetobacter calcoaceticus-Acinetobacter baumannii complex by matrixassisted laser desorption/ionization time-of-flight mass spectrometry / 中华检验医学杂志

Objective To evaluate the ability of matrix-assisted laser desorption/ionization time-offlight mass spectrometry (MALDI-TOF MS) in identifying species of Acinetobacter calcoaceticus-Acinetobacter baumannii complex,and investigate the species distribution of Acinetobacter calcoaceticus-Acinetobacter baumannii complex isolated in our hospital.Methods A total of 502 nonduplicate clinical isolates of Acinetobacter calcoaceticus-Acinetobacter baumannii complex were retrospectively collected from the second affiliated hospital of Zhejiang University between January 2012 and July 2012.All strains were re-identified by MALDI-TOF MS and were also verified by sequence analysis of 16S-23S rRNA gene spacer region.Results Among all the 502 strains of Acinetobacter calcoaceticus-Acinetobacter baumannii complex,identificaion results provided by MALDI-TOF MS were A.baumannii (431,85.9％),A.pittii (68,13.5％),A.calcoaceticus (3,0.6％).Sequence analysis of 16S-23S rRNA gene spacer region was used to identify all the 502 strains:403 (80.3％) were identified as A.baumannii,68 (13.5％) as A.pittii,28 (5.6％) as A.nosocomialis and 3 (0.6％) as A.calcoaceticus.MALDI-TOF MS correctly identified all the strains but erroneously identified all 28 strains of A.nosocomialis as A.baumannii,compared with sequence analysis of 16S-23S rRNA gene spacer region.Conclusions MALDI-TOF MS can be used as a fast,simple,reliable and excellently reproducible method to identify members of Acinetobacter calcoaceticus-Acinetobacter baumannii complex at low costs.MALDI-TOF MS is expected to be an ideal technique for routine clinical microbiology testing in the future.

The effect of different treatment programs on serum myocardial enzymes, amylase and C-reactive protein levels in patients with acute organophosphate poisoning / 中国医师进修杂志

ObjectiveTo explore the clinical efficacy of different treatment programs for patients with acute organophosphate poisoning (AOPP).MethodsOne hundred and thirty patients with AOPP were divided into treatment group (80 cases) and control group (50 cases) by table of random digit.The treatment group was given atropine intravenous injection combined with pralidoxime chloride ladder intramuscular injection on the basis of the conventional treatment.The control group was given atropine intravenous injection combined with pralidoxime chloride intravenous drip on the basis of conventional treatment.The healing rate,creatine kinase(CK),aspartate aminotransferase(AST),amylase and C-reactive protein (CRP) were compared between two groups.ResultsThe healing rates of patients with mild,moderate and severe poisoning patients in treatment group were respectively significantly higher than those in control group [ 100.00％(19/19) vs.75.00％(9/12),95.35％(41/43) vs.64.29％(18/28) and 88.89％(16/18) vs.60.00％ (6/10),P ＜0.05].There was no significant difference in CK,AST,amylase and CRP before treatment between two groups(P ＞ 0.05).The CK,AST,amylase and CRP after treatment in treatment group were significantly lower than those in control group [ ( 152.3 ± 23.2) U/L vs.(258.5 ± 22.2) U/L,(37.5 ± 11.4) U/L vs.(44.5± 12.4) U/L,(114.2±43.8) U/Lvs.(147.3 ±61.4) U/L,(5.7±4.1)mg/Lvs.(9.8±5.2)mg/L,P ＜0.05].Conclusions The use of atropine intravenous injection combined with pralidoxime chloride ladder intramuscular injection for mild,moderate and severe AOPP patients is excellent in therapeutic effects,and the clinical cure rates and blood biochemical parameters are more desirable.It is worthy of clinical application.

Comparison of antibacterial activities of different concentrations of the cefoperazone-sulbactam combination disc upon commonly found gram negative bacilli / 中华急诊医学杂志

Objective To evaluate antibacterial activities of Cefoperazone-Sulbactam upon gram negative bacilli,and compare the differences in susceptibility between two different concentrations of Cefoperazone-Sulbactam combination disc.Method A total of 381 strains of commonly occurred gram negative bacilli were found from 2nd Affiliated Hospital of Zhejiang University School of Medicine,Zhejiang Provincial People's Hospital,The Third Hospital of Hangzhou and Hangzhou Hospital of Traditional Chinese Medicine respectively.Susceptibility test was conducted by K-B method using 75 μg/disc Cefoperazone plus with 75 μg/disc Sulbactam(150 disc)and 75 μg/disc Cefoperazone with 30 μg/disc Sulbactam(105 disc),respectively.Meanwhile the minimal inhibitory concentration(MIC)of Cefoperazone-Sulbactam was determined by standard agar dilution.The data were analyzed by using WHONET 5.4 and SPSS.Results Disc diffusion method was carried out to detect the antibacterial activities upon Escherichia coli,Klebsiella pneumonia,Pseudomonas aeruginosa and Acinetobacter baumannii,using 105 disc and 150 disc,respectively.The data indicated that consistency rates between these two different discs were 26.3%,79.2%,83.7%and 33%,respectively.The k-related sample test was performed by using SPSS version 10.0 and shown that the P value was less than 0.05.Upon those organisms mentioned above,the consistency rates between the antibacterial activities of 105 disc and those of agar dilution were 77.8%,89.6%,70.9%and 77%,respectively.When it was going to compare agar dilution vs.150 disc upon the susceptibility of those organisms the consistency rates were 27.3%,79.2%,61.6%and 30%,respectively.Compared with agar dilution,the error rates of those two different concentration discs revealed that the false susceptibility and false intermediate of 105 disc were higher than those of 105 disc.ConclusionsThe results of susceptibility test showed that 105 disc was more close to agar dilution than that of 150 disc.However,the 150 disc used in clinic led to increase in sensitivity of susceptibility test to organisms.

Detection of AmpC ?-Lactamases and Analysis of Antimicrobial Activity in vitro for Extended Spectrum ?-Lactamases Positive Escherichia coli and Klebsiella pneumoniae / 中华医院感染学杂志

OBJECTIVE To detect AmpC ?-lactamases from isolates of extended spectrum ?-lactamases(ESBLs) positive Escherichia coli and Klebsiella pneumoniae,which were isolated from four hospitals in Hangzhou from 2005 to 2006,and to analyze the antimicrobial activity of clinically commonly used antibiotics in vitro.METHODS Amount of 324 ESBLs positive isolates including E.coli and K.pneumoniae were collected from Zhejiang Provincial People′s Hospital;1st Affiliated Hospital of Zhejiang University;2nd Affiliated Hospital of Zhejiang University and Hangzhou Traditional Chinese Medicine Hospital perspectivly.AmpC ?-lactamase was identified by disc screening testing and three dimensional test,and the genotypes of AmpC ?-lactamase were also determined by multiplex polymerase chain reaction(Multi-PCR).Minimal inhibitory concentrations(MICs) of ten clinically commonly used antibiotics were determined by agar dilution for AmpC ?-lactamases positive isolates,and the data were analyzed by WHONET 5.3.RESULTS AmpC ?-lactamase phenotype test revealed that 76 AmpC ?-lactamase positive isolates(23.5%) were identified among 324 ESBLs positive isolates of E.coli and K.pneumoniae from four hospitals in Huzhou.69.7% Of the AmpC ?-lactamase phenotype positive isolates were positively amplified by Multi-PCR.The value of MIC50 for carbapenem was lower 0.25 ?g/ml.We also found four carbapenem-resistant strains in this study.CONCLUSIONS We found that the incidence rate of AmpC ?-lactamase is high among the ESBLs producing E.coli and K.pneumoniae strains in Hangzhou.Carbapenem antibiotics have higher antimicrobial activity than other tested antibiotics.