Objective To analyze the antibiotic susceptibility, ESBL genotype of clinical Klebsiella pneumoniae strains isolated from People′s hospital and facilitate the control of resistance spread. Methods Identification and antibiotic susceptibility tests of 1 205 strains from 2001 to 2007 were done by VITEK-2 system.The antibiotic susceptibility results were analyzed by whonet5.3.The ESBL gene was detected by PCR and the Chi-square test was used for statistical analysis.Results The rate of ESBL-producing strains in klebsiella pneumoniae has increased from 2001 to 2007[18.8% (40/213) in 2001, 20.9% (53/253) in 2002, 32.8% (42/128) in 2003, 33.6% (45/137) in 2004, 36.6% (60/164) in 2005, 45.3% (68/150) in 2006 and 45.6% (73/160) in 2007].The SHV gene was the most dominant in ESBL genotypes.There were 83.3% (50/60) ESBL strains in 2005 with SHV gene, 82.3%(56/68) in 2006 and 83.6%(61/73) in 2007.The rated of strains with CTX-M gene were increasing.There were 26.7%(16/60) ESBL strains with CTX-M gene in 2005, 36.7%(25/68) in 2006 and 54.8%(40/73) in 2007.The isolates with more than one type of ESBL gene were increasing.There were 45%(27/60) ESBL strains in 2005 with two types of ESBL gene, and no one had more than two types of ESBL gene in that year.There were 47.9%(35/73) ESBL strains in 2007 with two types of ESBL gene.In 2007 there were 9.6%(7/73) and 2.7%(2/73) ESBL strains with three types and four types of ESBL gene respectively.There was a statistical difference between the antibiotic resistance rates of cefotaxime, ceftriaxone and ceftazidime in SHV-gene-phore strains (χ2=13.22, P＜0.01).The strains with SHV gene were more resistant to cefotaxime than ceftriaxone and ceftazidime.There also was a statistical difference of the antibiotic resistance rate of cefotaxime, ceftriaxone and ceftazidime between strains with TEM gene (χ2=9.91, P＜0.01) and CTX-M gene (χ2=34.84, P＜0.01) respectively.None of the strains with CTX-M gene was sensitive to cefotaxime, and they were more resistant to ceftriaxone than ceftazidime.The strains with TEM gene were more resistant to cefotaxime than ceftriaxone and ceftazidime.There were statistical differences of the antibiotic resistance rate to cefotaxime (χ2=29.65, P＜0.01), ceftriaxone (χ2=20.26, P＜0.01) and ceftazidime (χ2=20.26, P＜0.01) between the strains with SHV gene only and strains with SHV and CTX-M gene concurrently.There were also statistical differences of the antibiotic resistance rates to cefotaxime (χ2=11.01, P＜0.01), ceftriaxone (χ2=9.93, P＜0.01) and ceftazidime (χ2=7.01, P＜0.01) between the strains with SHV gene only and strains with SHV and TEM gene concurrently.The antibiotic resistance rates to cefotaxime (χ2=11.54, P＜0.01), ceftriaxone (χ2=17.58, P＜0.01) and ceftazidime (χ2=14.11, P＜0.01) were statistically different between the strains with SHV gene only and strains with SHV and OXA gene concurrently.The antibiotic resistance rates to ceftazidime (χ2=23.61, P＜0.01) were statistically different between the strains with CTX-M gene only and strains with SHV and CTX-M gene concurrently. There was no statistical difference in antibiotic resistance rates to cefotaxime (χ2=3.55, P＜0.01) and ceftriaxone (χ2=3.35, P＜0.01) between the strains with CTX-M gene only and strains with SHV and CTX-M gene concurrently. The antibiotic resistance rates to ceftazidime (P=0.01) were statistically different between the strains with only TEM gene and strains with SHV and TEM gene concurrently, and there was no statistical difference of the antibiotic resistance rates to cefotaxime (P=0.29) and ceftriaxone (P=0.26) between the strains with TEM gene only and strains with SHV and TEM gene concurrently. ConclusionsThe producing rate of ESBL is increasing year after year and the SHV type of ESBL is the dominant one.Strains with more than one type of ESBL gene are increasing.The antibiotic resistance rates to cefotaxime, ceftriaxone and ceftazidime are statistically different between strains with same ESBL genotype.

Objective:To evaluate the effects of intraarticular injection of bevacizumab、sodium hyalu-ronate (SH)and 0.9% sodium chloride injection in the treatment of osteoarthritis (OA)in a rabbit model.Methods:Twenty-four male rabbits were randomly divided into bevacizumab group,SH group and control group after the model of OA had been made.The bevacizumab group and control group received intraarticular bevacizumab (4 mg)and 0.9% saline injection respectively once per three weeks for 2 times.The SH group received intraarticular SH once a week for 6 weeks.After 6 weeks,the histological examinations of cartilage and synovium,electron microscopy and expression of vasculan endothelial growth factorl (VEGF),for the synovium,expression of MMP-1 ,Mankin’s scale,macroscopic observation for cartilage were performed.Results:The histological observation of the bevacizumab group and the SH group showed that bevacizumab could decrease the synoviocytes and inhibit fibrous hyperplasia in synovial underlayer compard with the control group.Reduced apoptosis of chondrocytes and more integrated struc-ture of matrix and more glycosaminoglycan were also found in the bevacizumab group and the SH group compared with control group.The expression of VEGF and MMP-1 ,Mankin’s scale,macroscopic obser-vation were significantly decreased in the bevacizumab group compared with the SH group and the control group (P<0.05).Conclusion:Intraarticular injection of bevacizumab and SH can relieve inflammation of OA and alleviate the pathologic process of OA.The Bevacizumab was better than the SH in therapeutic effect,which maybe implicate a better choice for the treatment of OA.

Objective To evaluate the toxicity of the intra-articular injection of bevacizumab in the knee of the rabbit.Methods Thirty-two rabbits were divided into 3 experimental groups and normal control group.Three experimental groups were received intra-articular injection of bevacizumab (1, 2, 4 mg respectively) once every three weeks for two times and the normal control group was received the same amount of 0.9% sodium chloride solution.The animals were sacrificed after 6 weeks.Blood test was examined before and after treatment.Pathologic examinations of liver, kidney and artiluar tissue were taken after the sacrifice.The hematoxylin and eosin stain for synovium and cartilage were performed.The AB-PAS stain and Mankin's scale for cartilage were performed.Results All the rabbits kept normal physiological activity.There was no significant difference of major organs and articular tissue between experimental groups and normal control group.There was no significant difference for WBC, RBC, PLT, ALT, BUN and Mankin's scale among all groups.Conclusion No systemic toxicity effects were found for the intra-articular injection of bevacizumab in the knee of the rabbit.

Objective To investigate the factors affecting the susceptibility of tigecycline and assess the testing methods.Methods The 116 isolates of Acinetobacter baumanaii and Klebsiella pneumoniae were collected in 13 hospitals from January to December,2010,to evaluate the effects on the tigecycline susceptibility of the overnight medium,medium brand and lot number,respectively.The 56 isolates of Acinetobacter baumannii and the 47 isolates of Klebsiella pneumoniae were selected randomly according to the MIC distribution proportion in 2010 and 2012.The broth microdilution was taken as the reference method to evaluate the effects of the agar dilution,disk diffusion,MIC Test Strip (MTS) and Vitek2 (GN16) on the susceptibility of tigecycline.Results The essential agreement (EA) of Acinetobacter baumannii and Klebsiella pneumoniae is 89.7％ (52/58)and 87.9％ (51/58) using overnight medium and fresh medium respectively.Both EA and categorical agreement (CA) of the different brands (BBL and Oxoid) and lot numbers are 100％ using agar dilution.According to the FDA break point criteria,the CA/EA is 77.7％ (80/103)/99.0％ (102/103),87.4％ (90/103)/98.1％ (101/103),64.1％ (66/103)/76.7％ (79/103) using agar dilution,MTS,Vitek (GN16) with respect to broth microdilution.The CA is 79.6％ (82/ 103,S≥14 mm,R≤10 mm),69.9％ (72/103,S≥ 16 mm,R≤ 12 mm),34.0％ (35/103,S≥19 mm,R≤14 mm)using disk diffusion method compared with broth microdilution (FDA break point criteria).Conclusions The susceptibility of tigecycline must be tested using fresh medium.The medium brands and lot numbers used in this test have no effects on the tigecycline susceptibility to Acinetobacter baumannii and Klebsiella pneumoniae.There exist the better correlations on MIC using agar dilution and MTS than the disk diffusion and Vitek(GN16) compared with broth microdilution.It is expected that the consistency can be improved by adjusting the break point of disk diffusion.

OBJECTIVE To surveillance invasive fungal infection rate in SICU,in order to direct intervention to prevent invasive fungal infection.METHODS The samples collected from SICU patients in our hospital between Jan 2003-Nov 2008 were cultured.RESULTS According to the diagnosis standard of nosocomial infections,75 case of 3699 patients were isolated fungi.During 6-years invasive fungal infection rate is 2.027%,(1.05%-2.63%).Totally 86 fungi strains were isolated,the majority of them being Candida albicans,accounting for 46.51%;Candida glabrata 22.09%;Candida tropicalis 13.95%.CONCLUSIONS During 6-years,invasive fungal infection rate and incidence density do not increase.Candida are the major pathogens of fungal infections in SICU.

Objective To investigate the diagnostic value of PIgP/SC in diagnosis of primary hepatic cancer.Methods 58 patients with primary hepatic cancer,60 patients with liver cirrhosis and 60 healthy volunteers were studied.4 ml fasting venous blood was collected from all subjects.Serum level of AFP was detected with electrochemical chemiluminescence immunoassay system and plasma PIgR/SC level was detected by ELISA method.The level of PIgR/SC and AFP was detected at one week after surgical resection in patients with hepatic cancer.Results The levels of AFP and PIgR/SC in the three groups were significantly different (P＜0.01),and PIgR/SC was higher than that in patients with cirrhosis and volunteers (P＜0.01).There was no significant difference between patients with liver cirrhosis and liver cirrhosis(P＞0.05).AFP was higher in patients with HCC than patients with cirrhosis and volunteers.AFP was higher in patients with cirrhosis than volunteers,and the difference was statistically significant(P＜0.01).Sensitivity of PIgR/SC and AFP was 89.3％ and 54.8％,specificity was 84.6％ and 91％,Youden index was 0.751 and 0.458,AUC was 0.920 and 0.761,respectively.There was significant difference in AUC (Z=3.251,P＜0.05) of the two detection indexes for detection of primary hepatic cancer.Conclusion The value of PIgR/SC in diagnosis of primary liver cancer may by higher than that of AFP.

Objective To elucidate the resistance mechanisms of clinical colistin-resistant Klebsiella pneumoniae and Escherichia coli isolates in China.Methods A total of 964 K.pneumoniae and 1 389 E. coli isolates were retrospectively collected from national surveillance programs from 2011 to 2014 in China. Antimicrobial susceptibility testing was determined by the microdilution method.The PCR amplification followed by sequencing was used to detect the mcr-1 gene and colistin-resistance genes, including mgrB, pmrB and phoQ.Real-time quantitative PCR was performed to examine the relative transcriptional levels of pmrB, pmrC, pmrD, pmrK and pmrE genes in K.pneumoniae, and pmrA, pmrB, pmrC, phoP and phoQ genes in E.coli.Conjugation experiment was used to detect the transferability of the resistance plasmid carrying the mcr-1 gene.Statistical analyses were performed using IBM SPSS Statistics (version 16.0) and a P value <0.05 was considered statistically significant. Results The colistin-resistant rates of K. pneumoniae and E.coli were 0.62% ( 6/964 ) and 1.66% ( 23/1 389 ) , respectively.No amino acids substitutions were identified in mgrB genes among colistin-resistant isolates.Among six colistin-resistant K. pneumoniae isolates, five isolates were identified to have point mutations in pmrB gene, but no point substitution was detected in phoQ gene.One to four point mutations had been found in pmrB and phoQ genes in colistin-resistant E.coli isolates, respectively.The expression level of pmrB, pmrC, pmrD, pmrK and pmrE genes showed no significant difference between colistin-resistant and colistin-susceptible isolates [pmrB, (1.04 ±1.12) vs.(0.94 ±0.67), P=0.945; pmrC, (1.39 ±2.01) vs.(0.16 ±0.27), P=0.101;pmrD, (1.59 ±2.43) vs.(0.88 ±0.34),P=0.445;pmrK, (0.64 ±0.62) vs.(0.04 ±0.10), P=0.051;pmrE, (3 492 833 388.83 ±8 478 977 986.85) vs.(20 771 428.93 ±38 000 732.85), P=0.445].However, the transcriptional level of pmrB genes in colistin-resistant group was 9.5-fold higher than that of the colistin-susceptible group in E.coli isolates.Four in six colistin-resistant K.pneumoniae isolates possessed mcr-1 gene, whereas all of the colistin-resistant E. coli had the mcr-1 gene. The conjugation verified the transferability rate of the plasmid carrying mcr-1 gene was 5.78 ×10-6 , and the MIC value of colistin of the conjugant increased 21-fold than the recipient strain.Conclusions Plasmid-mediated mcr-1 gene was the major reason for colistin resistance in clinical isolates of K.pneumoniae and E.coli. Some other resistance mechanisms such as transcriptional up-regulated pmrB gene also involved in colistin resistance.

Objective To evaluate the clinical performance of the BACTEC Plus aerobic,BACTEC Lytic anaerobic,BacT/Alert aerobic and anaerobic blood culture media in detection of bloodstream infections.Methods Retrospective study was conducted.A total of four blood culture bottles from each inpatient with suspected bloodstream infections were collected and analyzed from June 2013 to September 2015 in Peking University People's Hospital.The four bottles,including BACTEC Plus aerobic,BACTEC Lytic anaerobic,BacT/Alert FA aerobic and BacT/Alert FN anaerobic media,and was incubated for 5 days in the BacT/Alert 3D and BACTEC FX instruments,respectively.Time to detection (TTD) and positive rate in detecting bacteria of the two systems were evaluated by Wilcoxon test and Chi-square test.Results Among 2 189 total cultures collected,20 were excluded because of blood shortage and 201 (9.27％) were positive for pathogens.The positive rates of BACTEC Plus aerobic media and BacT/Alert FA aerobic media were 75.3％ (140/186) and 69.4％ (129/186) (x2 =1.625,P=0.202),respectively.While,the positive rates of BacT/Alert FN anaerobic media and BACTEC Lytic anaerobic media were 81.8％ (99/121) and 63.6％ (77/121) for total organisms,respectively (x2 =10.083,P =0.001).A significant difference in TTD was detected in BACTEC Plus aerobic media[11.0 (8.0-16.0) h] and BacT/Alert FA aerobic media[13.9 (10.4-18.7) h] (Z =-5.240,P ＜ 0.001).BACTEC Lyric anaerobic media[8.0(7.0-10.0) h] had a shorter TTD (Z =-4.299,P ＜ 0.001) than BacT/Alert FN anaerobic media[11.3(9.3-12.7) h].The positive rates of BACTEC and BacT/Alert system were 74.13％ (149/201) and 74.63％ (150/201),respectively,compared with taking one set from each system.Conclusions BACTEC media has a shorter TTD and almost the same bacterial recovery,and lower false positive rate than the BacT/ Alert media.

Objective To evaluate the efficacy of adenosine triphosphate ( ATP ) bioluminescence test on quality monitoring of flexible endoscope reprocessing. Methods The patient-used gastroscopes were randomly assigned to two groups after standard reprocessing by one nurse. The group A were assessed by ATP test followed by microbiological surveillance test after cleaning and disinfection; and the gastroscopes in group B were tested in a reverse order. The biological load after endoscopic cleaning and disinfection were compared between ATP test and microbiological surveillance, and the correlation of the two tests on quality assessing of endoscopic cleanliness and disinfection were analyzed. Results A total of 51 samples were valid. The qualified rate of cleaning was 45. 1％ ( 23/51) detected by ATP test, and the qualified rate of disinfection was 92. 2％ ( 47/51) detected by microbiological surveillance. There was no statistical difference in cleaning qualification rate ( 50. 0％ VS 40. 0％,χ2=0. 515, P=0. 473) and disinfection qualification rate ( 96. 2％ VS 88. 0％,χ2=0. 316, P=0. 574) between the two groups. The logarithmic value of relative light unit ( RLU) of ATP test after cleaning was higher than the value after disinfection ( 1. 68 ± 0. 50 VS 0. 88 ±0. 49, t=2. 335,P=0. 024) . For gastroscopes disinfected appropriately, the RLU logarithmic value of ATP test after cleaning ( t=2. 505, P=0. 016) and disinfection ( t=2. 485, P=0. 016) were lower than that of disqualification. The correlation between the value of ATP test and the colony forming units of microbiological surveillance after cleaning was not statistically significant ( r=0. 199, P=0. 171 ) , but there was a low correlation between the two monitoring results after disinfection (r=0. 391, P=0. 005). There was a low correlation between the value of ATP test after cleaning and the colony forming units of microbiological surveillance after disinfection ( r=0. 301, P=0. 032) . Conclusion ATP bioluminescence test can be used to evaluate the trend of endoscopic biological load change and the quality of flexible endoscope reprocessing.

Objective To analyze the immune function and shoulder function of sentinel lymph node biopsy combined with mastectomy in patients with early breast cancer.Methods A total of 80 patients with breast fibroadenoma were selected ,and they were randomly divided into surround -mammary areola incision group ( n=40 ) and conven-tional radial incision group(n=40) according to the digital table.The bleeding volume,operation time,hospital stay, satisfaction and curative effect were compared between the two groups .Results The markedly effective rate of surround-mammary areola incision group was 87.50％,which was significantly higher than that of the radial incision group(62.50％),and the difference was statistically significant (χ2 =10.400,P=0.001).The total effective rate was 100.00％in the surround -mammary areola incision group ,which was significantly higher than that in the radical incision group(χ2 =6.500,P=0.010).The time of operation,the amount of bleeding and the length of hospital stay in the surround-mammary areola incision group were (56.7 ±3.2) min,(62.1 ±1.8) mL and(8.3 ±0.8) d, respectively,which were lower than those in the radial incision group [(69.8 ±4.3)min,(71.1 ±2.4)min,(11.2 ± 1.3)d],there were statistically significant differences between the two groups (t=15.262,P=0.000;t=18.735, P=0.000;t=11.864,P=0.000).The incidence rates of breast milk symmetry,scar acceptance and adverse reaction in surround-mammary areola incision group were 87.50％,87.50％ and 2.50％,respectively,which were significantly better than those in the radial incision group(67.50％,65.00％ and 15.00％),there were statistically significant differences between the two groups (χ2 =5.032,P=0.024;χ2 =7.862,P=0.005;χ2 =3.923,P=0.047).The total satisfaction rate was 87.50％in the surround-mammary areola incision group ,which was higher than 62.50％in the radial incision group (χ2 =7.222,P=0.007).Conclusion The use of surround -mammary areola incision in the treatment of patients with breast fibroadenoma has obvious effect ,can reduce the degree of breast injury ,improve the physical appearance of the breast ,with high satisfaction .