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Objective:To analyze the characteristics of drug resistance genes in a Klebsiella pneumoniae strain coproducing carbapenemases KPC-2 and NDM-5. Methods:Klebsiella pneumoniae KPN-hnqyy was separated from the stool specimen of a patient in the Hematology Department of Affiliated Cancer Hospital of Zhengzhou University. The strain was identified with a BD Phenix-M50 automated microbiology system and the minimum inhibitory concentration against the strain was measured as well. The genotypes of the carbapenemases were tested by enzyme immunochromatographic assay and PCR method. The transferability of related plasmids was analyzed by conjugation test. Whole-genome sequencing of the strain was conducted using PacBio and Illumina platforms. The MLST type, resistance gene and plasmid type of the strain were retrieved in BacWGSTdb. The genome and open reading frame sequence of the strain were compared using Easyfig_2.2.3. Visual cycle graphs were generated using BRIG v0.95. Results:Klebsiella pneumoniae KPN-hnqyy was resistant to carbapenem antibiotics. It belonged to ST11 and carried two carbapenemase genes of blaKPC-2 and blaNDM-5. The conjugant only harbored the blaKPC-2 gene. Whole-genome sequencing revealed that the strain contained one chromosome and three plasmids. Its chromosome genome shared more than 99.9% similarity with that of Klebsiella pneumonia KP69 and KP19-2029. Moreover, a similar IncR and IncFⅠ resistance gene fusion region was contained in different types of plasmids carried by them: the blaKPC-2 gene was located in a structure—which evolved from the Tn3-△Tn4401-Tn1721/Tn1722 sequence—inside this fusion region with its ends inserted into the transposase IS26 gene; the blaNDM-5 gene was located on a transposon containing the special plasmids of the insertion fragment in phages, with its ends inserted into the transposase IS26 gene too. Conclusions:The IncR and IncFⅡ resistance gene fusion region of blaKPC-2 carried by Klebsiella pneumoniae ST11 might be widely coexistent with the chromosomal genome. The blaNDM-5 gene carried by special plasmids might be accidentally obtained through gene recombination mediated by transposable element IS26. The wide transmission of Klebsiella pneumoniae ST11 carrying the blaKPC-2 gene in China and its ability to obtain other carbapenemase genes through transposable element IS26 were well worth attention.
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Objective:To analyze the clinical characteristics of primary acute myeloid leukemia (AML) (non-M3 type) in children suffering from different levels of platelet count(PLT).Methods:In the Tumor Hospital of Zhengzhou University from January 2014 to December 2018, laboratory and clinical data of 247 de novo primary AML pediatric patients were retrospectively reviewed.According to the PLT before treatment, patients were divided into very low platelet group (VLG), low platelet group (LG) and non-lowing platelet group (NLG), with<50×10 9/L, ≥50×10 9/L but <125×10 9/L and ≥125×10 9/L as the boundaries.All patients were followed up until June 30, 2019.Meanwhile, the follow-up data was obtained by consulting medical records or by telephone.SPSS 17.0 software was applied for data analysis. Results:In general clinical features, a different group of hemoglobin (Hb) content, fusion gene AML- ETO and clinical risk stratification were statistically significant in different PLT groups ( χ2=11.270, 12.115 and 12.848, respectively, all P<0.05). However, the differences of other indicators in different groups of PLT were not statistically significant (all P>0.05). There were no statistically significant differences in terms of 3-year disease-free survival(DFS) rate (59.3%, 36.3%, 50.4%) among the 3 groups (all P>0.05). The median total survival(OS)time(40.5 months)and 3-year OS rate(41.0%) of NLG patients were significantly higher than those of VLG(23.1 months, 30.1%)and LG(14.1 months, 18.2%)patients, with statistically significant differences( χ2=7.798 and 6.553, respectively, all P<0.05). The univariate analysis of gender, white blood cell(WBC), Hb, PLT, lactic dehydrogenase(LDH), FLT3-ITD, NPM1, DNMT3A, CEPBA, C-KIT, AML-ETO, molecular genetic prognosis, complete remission(CR), and hemopoietic stem cell transplantation(HSCT) displayed that DNMT3A mutation was an adverse factor that affects patients′ OS ( χ2 =5.834, P<0.05), and the positive factors that influences OS were non-reducing PLT before treatment, and obtaining CR and subsequent HSCT ( χ2=7.798, 79.168, and 31.337, respectively, all P<0.05). Multi-factor analysis revealed that the independent protective factors that affect patients′ OS were the non-reducing PLT before treatment, and obtaining CR and subsequent HSCT( Wald=42.760, 15.918, and 10.183, respectively, all P<0.05). Conclusions:Before treatment, non-reducing PLT is a protective factor for primary childhood AML patients, and the prognosis is satisfying.
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Objective:To analyze the characteristics of plasmids in KPC-2-producing Serratia marcescens ( S. marcescens) isolates. Methods:Four carbapenem-resistant S. marcescens strains were isolated from four patients admitted to the hepatobiliary ward of Affiliated Cancer Hospital of Zhengzhou University in 2016. BD Phenix-100 was used to identify the strains and detect the minimum inhibitory concentrations (MICs). Homology analysis was performed using pulsed-field gel electrophoresis (PFGE). The modified Hodge test was used to detect the phenotypes of carbapenemase. PCR and gene sequencing were used to detect the types of carbapenem resistance genes. The transferability of plasmids was detected by conjugation test. The characteristics of plasmids were analyzed by genomic alignment method after whole genome sequencing. DNAMAN V9 software was used to compare the amino acid sequences of the replication initiation proteins. A phylogenetic tree was constructed with neighbor-joining method using MEGA7.0. Results:All of the four S. marcescens strains were resistant to carbapenem antibiotics. They were highly homologous according to PFGE. Hodge test results were all positive and the carbapenemase genotype was blaKPC-2. Conjugation test results were positive. The plasmid was a circular DNA of 42 742 bp in length. It had the similar skeleton of incX6 plasmid and the similar amino acid sequence of replication initiation protein. Moreover, it and incX6 plasmid were at the same node in the phylogenetic tree. The blaKPC-2 was located in the core of drug resistance, which was composed of insertion elements including Tn3 family transposons, recombinant enzyme genes, △ISKpn6 and ISKpn27. Conclusions:The plasmid was incX6-like. The blaKPC-2 gene was located in the transposon of △Tn6296. More attention should be paid to the bacteria carrying KPC-2 in incX plasmids.
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Objects: To clarify the risk factors of candidemia and to assess the clinical differences that may exist between infection with Candida parapsilosis and that with other Candida species in cancer patients. To statistically analyze the clinical characteristics of Candi-da albicans candidemia and C. parapsilosis candidemia and risk factors for their infections. We aimed at a timely intervention through this type of analysis to avoid susceptible factors and improve the prognosis of patients with candidemia. Methods: We retrospectively included 323 patients with candidemia in Affiliated Cancer Hospital of Zhengzhou University between March 2012 and February 2018 and analyzed the clinical characteristics of these patients to establish the risk factors of candidemia. We performed a comparative anal-ysis of the clinical characteristics of C. parapsilosis infections and non-parapsilosis Candida spp. infections and of C. albicans infections and non-albicans Candida spp. infections. In addition, drug sensitivity tests and analyses were performed with the common antifungal drugs used in Candida infections by a micro-broth dilution method. The statistical software SPSS version 22 was used for the analyses. Results: A total of 323 patients were enrolled and analyzed in this study. Of the isolates, 34.37% were C. albicans and 65.63% were non-albicans Candida spp. Multivariate regression analysis showed that the following factors were associated with the occurrence of C. parapsilosis candidemia: parenteral nutrition (P<0.001), neutropenia (P<0.001), history of receiving chemotherapy (P=0.002), and history of previous antifungal use (P<0.001). Parenteral nutrition was found to be an independent risk factor for C. albicans candi-demia (OR=0.183; 95%CI:0.098?0.340; P<0.001). Conclusions: C. parapsilosis was found to be the primary pathogen in cancer patients with candidemia. Total parenteral nutrition in the intensive care unit at diagnosis and abdominal surgery were independent risk factors of candidemia, and parenteral nutrition was an independent risk factor of C. parapsilosis candidemia. At present, C. parapsilosis is sur-passing C. albicans as the main pathogen of candidemia in cancer patients at our hospital. This study emphasizes the need to assess the possible risk factors for candidemia in cancer patients and aims at strengthening and developing a hospital-based control strategy to prevent the spread of candidemia.
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Objective To analyze the clinical characteristics of de novo primary AML patients with higher blast cell ratio in peripheral blood than bone marrow and its relationship with CR1. Methods The clinical data on de novo primary AML patients in Henan Provincial Tumor Hospital from January 2015 to December 2016 were ret-rospectively reviewed. Based on the proportion of blast cells in the peripheral blood and bone marrow ,the patients were divided into BHG and NBHG. All the data was analyzed respectively by Schisquare test ,rank sum test or Spearman correlation analysis according to the types of clinical data. Results As compared with NBHG patients , BHG patients had a higher rate of bleeding,palpitation,M2 subtype,FLT3-ITD mutation,and average level of LDH andα-HBDH in serum before treatment,and the difference was statistically significant(P<0.05);however, the rate of CR1,M3 and M5 subtype in BHG patients was lower than that in NBHG patients,and the difference was statistically significant(P < 0.05). The proportion of peripheral blood blast cells in patients with AML has a positive correlation with serum levels of LDH and α-HBDH(R:0.331 and 0.352,P < 0.05). Conclusions De novo primary AML patients with higher blast cell ratio in peripheral blood than bone marrow are mostly M2 subtype,easily associated with FLT3-ITD mutation,bleeding and palpitation symptoms,and they have a lower CR1rate. The proportion of blast cells in peripheral blood is positively correlated with levels of serum LDH and α-HBDH.
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Objective Combined with the medical records of two patients with megaloblastic anemia (MA) after allogeneic hematopoietic stem cell transplantation (AHSCT),the relevant literature was reviewed.Methods The medical records of two patients with MA after AHSCT were analyzed retrospectively.The primary disease was diagnosed by analyzing the blood cells,bone marrow cell morphology,cell chemical dyeing,bone marrow biopsy and immune classification.After AHSCT,MA was diagnosed through bone marrow cell morphology,folic acid and vitamin B12 detection.Results AT 32nd day after transplantation,bone marrow cells morphological examination of case 1 showed:nucleated cells proliferation activity,granulocytes proliferation activity,giant rod nucleus granulocytes visible;different stages of the red blood cells proliferation activity,higher proportion of immature red blood cells,most of whose nucleus developed later than the cytoplasm;megakaryocytes and platelets scattered distribution.Blood contents of folic acid and VB12 were far below the reference range.After administration of folic acid and VB12 for 2 weeks,routine blood test showed the volume of red blood cells returned to normal.Reexamination of bone marrow cell morphology showed megaloblastic cells disappeared.Three months after transplantation,bone marrow cells morphological examination in case 2 showed:nucleated cells proliferation activity,low granulocytes proliferation,giant rod nucleus granulocytes visible;different stages of the red blood cells proliferation activity,higher proportion of orthochrmatic normoblasts,most of whose nucleus developed later than the cytoplasm;scattered distribution of megakaryocytes and platelets.Blood contents of folic acid were far below the reference range,but the content of VB12 was normal.After administration of folic acid and VB12 for 2 weeks,the routine blood test showed the volume of red blood cells returned to normal.The reexamination of bone marrow cell morphology showed megaloblastic cells disappeared.Conclusion After AHSCT,attention should be paid to the detection of folic acid and VB12 in vivo.Folic acid and VB12 are timely supplemented when necessary to avoid the occurrence of MA in patients with AHSCT.
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Objective To study plasmid-mediated transfer,plasmid replicon typing,and genetic environment of blaNDM-1 gene in Enterobacteraerogenes(E.aerogenes).Methods E.aerogenes HN-NDM0711 was used as the subject of this research,the transferable properties of plasmid were analyzed by conjugation testing,conjugant was performed stability testing,plasmid type was determined by PCR-based replicon typing (PBRT),downstream and upstream of blaNDM-1 were sequenced using chromosome walking method,genetic context was analyzed by BLASTN and BALSTP,as well as annotated using Vector NTI 11.5.1 software,sequence pipeline graph was made,the sequence was submitted to Genbank through software Banklt.Results The conjugation testing of E.aerogenes pHN-NDM0711 was positive,after positive conjugant was conducted 4-day passage,minimal inhibitory concentrations (MICs) of imipenem and meropenem to all the cloned strains didn't change,blaNDM-1 were all positive.The replicon type was IncA/C;blaNDM-1 gene was localized between ISAba14 and IS91,at upstream of the blaNDM-1,class 1 integron and Tn3 transposon were identified,class 1 integron contained a new mosaic structure of a drug-resistant resistance gene cassette.Conclusion E.aerogenes pHN-NDM071 1,bearing blaNDM-1 gene in IncA/C plasmid,derived from gene recombination under different antimicrobial selection pressure.Antimicrobial use in clinical,industrial and agricultural area should be strictly controlled,so as to reduce the emergence of such bacteria.
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Objective To investigate the difference of clinical features between FLT3-ITDmt and FLT3-IT-Dwt de novo primary acute myeloid leukemia(AML). Methods Clinical data of 31 FLT3-ITDmt and 113 FLT3-ITDwt de novo primary AML patients from January 2015 to December 2016 were retrospectively reviewed and ana-lyzed by Student′s test,chi-square test or rank sum test according to the types of clinical data. Results There were statistically significant differences in WBC,RBC,HGB of peripheral blood and the mutation of DNMT3A gene(statistical values:705.000;-2.535;-2.290 and 5.715 respectively,all P < 0.05)in 2 types of AML. Conclusion When compared with FLT3-ITDwt de novo primary AML patients,FLT3-ITDmt ones have the fea-tures of higher WBC,lower RBC and HGB of peripheral blood,and are more likely to be associated with mutation in the DNMT3A gene.
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Objective To investigate distribution characteristics of blood culture pathogens,and provid a basis for clinical prevention and control in bloodstream infections.Methods The data of the patients with positive blood culture and the nonduplicate strains were retrospectively analyzed with WHONET 5.6 software in the First People's Hospital of Shangqiu from January 2011 to December 2014.Results The total number of positive strains of blood culture was 1 306 from 25 397 blood cultures,and the positive rates were 5.1％,of which gram positive cocci and gram negative organisms accounted for 64.8％ (846/1 306) and 31.2％ (408/1 306),respectively.Candida accounted for 4.0％ (52/1 306).Gram positive bacteria were mainly Coagulase negative Staphylococcus (CNS) of 564 (43.2％) strains,Staphylococcus aureus of 96 (7.4％) strains,Enterococcus faecium of 48 (3.7％) strains and Streptococcus pneumoniae of 31 (2.4％) strains.The isolated rate of Enterococcus faecium was more than Enterococcus faecalis 20 strains (1.5％).The isolated rate of Escherichia coli,Klebsiella pneumoniae,Acinetobacter Bauman and Pseudomonas aeruginosa were 12.5％ (163 strains),6.2％(81 strains),2.0％(26 strains) and 1.8％(23 strains),respectively.Conclusion The isolated rate of Enterococcus faecium more than Enterococcus faecalis in blood culture,the main pathogens are gram positive cocci in children group.Isolates of gram positive bacteria in the proportion of infants,children and adults with blood culture were 85.2％,87.0％,46.5％.There were 264 strains bacteria isolated in infants and young children's intensive care unit,accounting for 67.3％ of all bacteria isolated from infants and young children.There were 122 strains bacteria isolated in pediatric intensive care unit,accounting for 56.7％ of all bacteria isolated from childen.There were 255 strains bacteria isolated in adult intensive care unit,accounting for 36.5％ of all bacteria isolated from adult.Severe basic diseases were the vulnerable groups of bloodstream infections.
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Objective To investigate the distribution and drug resistance of major pathogens for urinary tract infections and related risk factors of hospitalized patients for prevention and treatment of urinary tract infections and to guide the reasonable application of antibiotics.Methods From January 2007 to December 2013 ,the risk factors and pathogen resistance of hospitalized patients with urinary system infection in the First People' s Hospital of Shangqiu were analyzed.Results Totally 891 strains were isolated, in which 638 (71.6％) were Gram-negative strains, 218 (24.5％) were Gram-positive strains, and 35 (3.9％) were Candida.Among Gram-negative strains, Escherichia coli was the most prevalent species, accounting for 43.5％, followed by Enterococcus spp(22.1％), major strains were Enterococcus faecalis and Enterococcus faecium.About 62.1％ (553/891) of the strains were isolated from female patients, 37.9％ (338/891) from male patients.The prevalence of ESBLs-producing strains in Escherichia coli, Klebsiella spp and P.Mirabilis were 59.0％ (526/891), 57.8％ (515/891) and 15.1％ (135/891).ESBLs-producing Enterobacteriaceae strains were more resistant than non-ESBLs-producing strains in terms of antibiotic resistance rate.The ESBLs-producing Enterobacteriaceae strains were highly susceptible to carbapenems.The average prevalence of Pseudomonas aeruginosa was 4.0％(36/891).Six strains were identified as multi-drug resistance of Pseudomonas aeruginosa were respectively, and 3 were carbapenem-resistant strains.Enterococcus spp were completely sensitive to linezolid, vancomycin and teicoplanin.Among Candida strains,the major strains Candida albicans was completely sensitive to amphotericin B, amphotericin and nystatin.The percentage of Candida albicans resistant to itraconazole,voriconazole and fluconazole was 13.0％ (116/891), 17.4％ (155/891) and 39.1％ (348/891).Clinical urinary tract infections related to sex, age, basic diseases, invasive operation and unreasonable application of antibiotics significantly (P＜0.05).Conclusion Escherichia coli is the leading clinical isolates strains account for most urinary tract infections in the First People' s Hospital of Shangqiu, followed by Enterococcus spp.The prevalence of Candida albicans is the major strains among Candida strains.Should be pay attention to the susceptible population and apply anbiotics reasonably.
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Objective To evaluate the protective effect of Bletilla striata polysaccharide ( BSPS) on immunological and chemical liver injury in mice. Methods Thirty Kunming male mice were randomly divided into five groups, including the normal control group,model control group,and low-,middle-,and high-dose BSPS groups (n=6 each).Tail vein injection of ConA was carried out to establish the ConA-induced liver injury model.After different treatments,all the animals were sacrificed,and the plasma levels of ALT and AST were tested.Additionally,sixty Kunming male mice were randomly divided into six groups,including the normal control group,model control group,silymarin group,and low-,middle-,and high-dose BSPS groups (n=10 each).Tail vein injection of CCl4 was performed to establish the CCl4-induced acute liver injury model.After different treatments,the plasma levels of ALT and GSH were tested.The effects of BSPS on the weights of the liver and spleen were examined. Results The levels of ALT and AST were reduced in BSPS-treated mice when compared with those experiencing only ConA-induced liver injury ( model control group) ,and significant difference was found between the middle-and high-dose BSPS groups and the model control group (P<0.01,P<0.05).The weights of the liver and spleen and the level of ALT were reduced in BSPS-treated mice as compared with those with only CCl4-induced acute liver injury (model control group),while the level of GSH was significantly increased in middle-and high-dose BSPS groups (P<0.05). Conclusion BSPS at low,middle,and high doses can prevent against the ConA-induced immunological liver injury and CCl4-induced acute liver injury in mice.
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Objective To investigate the dynamic changes and drug resistance of fungal infection in our hospital during 2010 -2012 for guiding the clinical reasonable use of anti-fungal drugs .Methods The related clinical data and the drug sensitivity tests re-sults of fungal infection in the hospitalized patients from 2010 to 2012 were retrospectively analyzed by the WHONET 5 .6 soft-ware .Results Candida albicans was the major pathogen causing candida infection for these 3 years .The isolated specimens were mainly sputum and blood .In the departments of internal medicine ,general surgery and integrated Chinese and Western medicine ,the number of detected candida were more than that of the other departments .Five kinds of candida showed high sensitivity to ampho-tericin B and high resistance to itraconazole .Conclusion The distribution of pathogens causing nosocomial fungal infection has changed and the drug resistance rate is continuously rising .Therefore the dynamic monitoring and the study of fungal infection should be strengthened for reducing the occurrence of fungal infection in the patients with tumor .
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ObjectiveTo investigate the flora distribution and drug resistance status of nosocomial fungal infection in cancer hospitals for providing the scientific basis for clinical prevention and control.Methods The related clinical data and results of drug sensitivity tests of fungal infection in hospitalized patients from April 2006 to November 2010 were analyzed retrospectively.ResultsA total of 1011 strains of candida were isolated in five years, in which, Candida albicans (41.6％ on average) were the highest detection rate strains.The proportion of Candida albicans declined from 53.8％ in 2006 to 31.7％ in 2010, while the other non-Candida albicans increased from 46.2％ in 2006 to 68.3％ in 2010.In the General surgery, the Internal medicine,the Hepatobiliary surgery, and the Chinese and Western medicine, the number of candida were more than the other departments.The specimens were mainly isolated from sputum and swab, followed by blood specimens.In vitro susceptibility test results showed that: voriconazole, amphotericin B and 5-fluorocytosine showed high sensitivity and itraconazole, fluconazole showed high drug resistance to albicans.Conclusion Distribution of pathogens causing nosocomial fungal infection has changed, and the rates of fungal resistance were rising,so this situation should arouse the clinician's attention.
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OBJECTIVE To study the incidence of ?-lactamases,mainly the extended-spectrum beta-lactamases(ESBLs) and metallo-beta-lactamases(MBLs) of Chryseobacterium indologenes and Ch.gleum.METHODS Agar dilution method was applied to detect minimal inhibitory concentrations(MIC) to 12 different antibiotics used frequently.Three-dimensional test was used to detect ESBLs and metallo-?-lactamases.The genes of ?-lactamases were amplified with 3 pairs of primers special for Ch.indologenes and Ch.gleum.RESULTS Among the 25 strains of Ch.indologenes and 10 strains of Ch.gleum,68%(17/25) isolates of Ch.indologenes and 90%(9/10)isolates of Ch.gleum were considered as MBLs positive strains,but no isolates were detected for the production of ESBLs.CONCLUSIONS MBLs are the important mechanism of multi-drug resistance for Ch.indologenes and Ch.gleum.