1.System analysis of the ecological distribution of bacteriophages in hospital wastewater
Jianying LU ; Hongwei PAN ; Enhua SUN ; Wei LI ; Hongxing WANG ; Xiulan ZHAO ; Hongchun WANG
Chinese Journal of Preventive Medicine 2024;58(2):241-247
Phage therapy is one of the most important tools for the treatment of infections with multi-drug resistant bacteria. Such phages are usually isolated from hospital effluents, however, no systematic study on the distribution of phages in hospital effluents has been conducted so far. The aim of this study was to isolate the corresponding phages of common pathogenic bacteria isolated in the clinic as hosts, so as to assess the ecological distribution of phages in hospital wastewater and to provide a reference for the isolation and application of phages of drug-resistant bacteria in the clinic. A cross-sectional study design was used in this study. The 125 pathogenic bacteria (belonging to 16 different strains) isolated from the clinical microbiology laboratory of Qilu Hospital of Shandong University from May to June 2023 were selected as the target strains, and the phages corresponding to these strains were isolated and purified from the hospital wastewater by using the double-layer plate sandwich method. At the same time, the distribution of pathogenic bacteria in the same batch of wastewater was analyzed with the help of mNGS sequencing technology, so as to preliminarily investigate the abundance correspondence between pathogenic bacteria and phages in wastewater. The results showed that a total of 56 phage strains were isolated from 125 clinical pathogens as hosts, corresponding to six pathogens, including Acinetobacter baumannii, Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia. All six pathogenic bacteria contained strains with different degrees of drug resistance, with a higher percentage of multi-drug resistant strains in A. baumannii, Escherichia coli and P. aeruginosa. The phage acquisition rates of these six pathogens were, in descending order, Escherichia coli (80%), Stenotrophomonas maltophilia (75%), Pseudomonas aeruginosa (70%), Klebsiella pneumoniae (66.67%), Acinetobacter baumannii (36.36%) and Staphylococcus aureus (12.5%). Preliminary mNGS sequencing results showed that the pathogenic bacteria with higher abundance in the batch of effluent were Enterococcus faecalis, Klebsiella pneumoniae, Escherichia coli, Enterococcus faecalis, Acinetobacter baumannii, Klebsiella aerogenes, Klebsiella michiganensis and Pseudomonas aeruginosa. In conclusion, phages of most common clinical Gram-negative pathogens were isolated from hospital wastewater with high isolation rates; however, phages of Gram-positive pathogens were isolated at lower rates, and only phages corresponding to Staphylococcus aureus were isolated in this study. The corresponding mNGS sequencing results showed that the distribution of Gram-negative pathogens in sewage may had a positive correlation with the ecological distribution of phages.
2.System analysis of the ecological distribution of bacteriophages in hospital wastewater
Jianying LU ; Hongwei PAN ; Enhua SUN ; Wei LI ; Hongxing WANG ; Xiulan ZHAO ; Hongchun WANG
Chinese Journal of Preventive Medicine 2024;58(2):241-247
Phage therapy is one of the most important tools for the treatment of infections with multi-drug resistant bacteria. Such phages are usually isolated from hospital effluents, however, no systematic study on the distribution of phages in hospital effluents has been conducted so far. The aim of this study was to isolate the corresponding phages of common pathogenic bacteria isolated in the clinic as hosts, so as to assess the ecological distribution of phages in hospital wastewater and to provide a reference for the isolation and application of phages of drug-resistant bacteria in the clinic. A cross-sectional study design was used in this study. The 125 pathogenic bacteria (belonging to 16 different strains) isolated from the clinical microbiology laboratory of Qilu Hospital of Shandong University from May to June 2023 were selected as the target strains, and the phages corresponding to these strains were isolated and purified from the hospital wastewater by using the double-layer plate sandwich method. At the same time, the distribution of pathogenic bacteria in the same batch of wastewater was analyzed with the help of mNGS sequencing technology, so as to preliminarily investigate the abundance correspondence between pathogenic bacteria and phages in wastewater. The results showed that a total of 56 phage strains were isolated from 125 clinical pathogens as hosts, corresponding to six pathogens, including Acinetobacter baumannii, Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia. All six pathogenic bacteria contained strains with different degrees of drug resistance, with a higher percentage of multi-drug resistant strains in A. baumannii, Escherichia coli and P. aeruginosa. The phage acquisition rates of these six pathogens were, in descending order, Escherichia coli (80%), Stenotrophomonas maltophilia (75%), Pseudomonas aeruginosa (70%), Klebsiella pneumoniae (66.67%), Acinetobacter baumannii (36.36%) and Staphylococcus aureus (12.5%). Preliminary mNGS sequencing results showed that the pathogenic bacteria with higher abundance in the batch of effluent were Enterococcus faecalis, Klebsiella pneumoniae, Escherichia coli, Enterococcus faecalis, Acinetobacter baumannii, Klebsiella aerogenes, Klebsiella michiganensis and Pseudomonas aeruginosa. In conclusion, phages of most common clinical Gram-negative pathogens were isolated from hospital wastewater with high isolation rates; however, phages of Gram-positive pathogens were isolated at lower rates, and only phages corresponding to Staphylococcus aureus were isolated in this study. The corresponding mNGS sequencing results showed that the distribution of Gram-negative pathogens in sewage may had a positive correlation with the ecological distribution of phages.
3. Method selection for pretreatment in direct identification of pathogen in positive blood culture by MALDI-TOF MS
Chinese Journal of Laboratory Medicine 2018;41(8):563-566
Bloodstream infection is one of the leading causes of death worldwide. Rapid determination of the primary pathogen is crucial for diagnosis and antibiotic therapy of bacteremia. The application of Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) was a major breakthrough in microbiology. This technology is simple, rapid, economical and accurate, which has been introduced into the microbiology laboratory formicroorganism identificationwidely. Recently, MALDI-TOF MS was combined with extraction methods for direct pathogens identificationfrom positive blood cultures. The methods of pathogensextraction from positive blood cultures are crucial for the accuracy of identification. And developing a standard extraction protocolhas become a hot spot of research in recent years. (
4.Optimum cuff pressure of flexible laryngeal mask airway for airway management in pediatric patients
Jun LUO ; Ruiqiang SUN ; Enhua GU ; Yuliang XUE
Chinese Journal of Anesthesiology 2017;37(2):214-217
Objective To determine the optimum cuff pressure of the flexible laryngeal mask airway (LMA) for airway management in pediatric patients.Methods One hundred and twenty pediatric patients undergoing strabismus surgery with general anesthesia,of American Society of Anesthesiologists physical status Ⅰ or Ⅰ,aged 3-10 yr,were randomly divided into 3 groups (n=40 each) according to the cuff pressure of the flexible LMA:20 cmH2O pressure group (group A),40 cmH2O pressure group (group B) and 60 cmH2O pressure group (group C).The cuff was inflated to the predetermined pressure using the inflatable cuff manometer in each group.Oropharyngeal leak pressure was measured after LMA placement.Peak airway pressure and the difference between inhaled and exhaled tidal volume were recorded at 5 min of positive pressure ventilation.The LMA insertion condition,LMA removal time,and development of sore throat,hoarseness,dysphagia and abdominal distention within 24 h after operation were recorded.Results There were no significant differences in the success rate of LMA placement at first attempt,peak airway pressure,or incidence of sore throat among the three groups (P>0.05),and no hoarseness or dysphagia was found in the three groups.Compared with group A,oropharyngeal leak pressure was significantly increased,and the difference between inhaled and exhaled tidal volume and incidence of abdominal distention were decreased in B and C groups (P<0.05).There was no significant difference in the parameters mentioned above between group B and group C (P>0.05).Conclusion The optimum cuff pressure of the flexible LMA is 40 cmH2O when used for airway management in the pediatric patients.
5.Consensus of Chinese experts on detection of related drive genes in target therapy of non-small cell lung cancer.
Enhua WANG ; Minghua ZHU ; Hong BU ; Jie CHEN ; Yanqing DING ; Xiaojun ZHOU ; Zhiyong LIANG ; Xianghong ZHANG ; Baocun SUN ; null ; null ; null ; null
Chinese Journal of Pathology 2016;45(2):73-77
6.The Effects of Remifentanil on Spontaneous Ventilation in Children Received Sevoflurane Anesthesia
Jun LUO ; Ruiqiang SUN ; Yongwang WANG ; Enhua GU
Tianjin Medical Journal 2014;(9):933-936
Objective To investigate the effects of different infusion rates of remifentanil infusion on spontaneous ventilation in children received sevoflurane anesthesia. Methods A total of 120 children underwent strabismus surgery were randomly assigned to four groups: C group (administration of saline), L group (remifentanil 0.03 μg · kg-1 · min-1), M group (remifentanil 0.06μg · kg-1 · min-1) and H group (remifentanil 0.09μg · kg-1 · min-1). The mean blood pressure (MBP), heart rate (HR), respiratory rate (RR), tidal volume (VT), minute ventilation (MV), endtidal CO2 [p(CO2)] and endtidal SEV were recorded after laryngeal mask insertion (T1), an initial bolus dose of remifentanil (T2),10 mins after remifentanil infu-sion (T3),15 mins after remifentanil infusion (T4) and laryngeal mask remove (T5) respectively. The adverse events and time of induction, maintenance and emergence were also recoded. Results There were no significant differences in patient age, body mass index, anesthesia time, operation time, HR and MBP at different time points between four groups. No body movement and hypoxemia were observed. The values of RR and MV at T3, T4 and T5 were significantly lower in H group than those of other three groups (P <0.05). Values of p(CO2)at T3 and T4 were significantly higher in H group than those of other three groups (P<0.05). The values of RR at T3, T4 and T5 were significantly lower in L group and M group than those of C group. The values of MV at T3 and T4 were significantly lower in L group and M group than those of C group. p(CO2)at T4 was significantly higher in L group and M group than that of C group(P<0.05), but no significant difference was found be-tween L group and M group. There was no significant difference in value of VT between four groups. Conclusion Remifent-anil infusion at a rate of 0.03~0.09μg·kg-1·min-1 could depress spontaneous ventilation in children received sevoflurane an-esthesia. The respiratory depression effect is mainly manifested by reduction of RR. It is a good option to choose 0.03~0.06μg · kg-1 · min-1 infusion to keep spontaneous ventilation and avoid severe respiratory depression according to the demand of operations in children.
7.MR subtraction in the differentiation of benign and malignant tumors.
Jianning SUN ; Dujun BIAN ; Enhua XIAO ; Weijun SITU ; Zhong HE ; Huabing LI ; Shuwen YUAN
Journal of Central South University(Medical Sciences) 2013;38(10):997-1002
OBJECTIVE:
To evaluate dynamic contrast-enhanced MRI in conjunction with MR subtraction in the differential diagnosis of benign and malignant breast tumors at 3 Tesla.
METHODS:
A total of 78 patients with breast tumors enrolled in this study, including 45 malignant lesions and 33 benign lesions verified by histopathology. Dynamic MR contrast enhanced imaging was done by T1 high resolution isotropic volume excitation sequence. MR subtraction was used to retrospectively analyze the MR dynamic image. A dynamic phase subtraction (DPS) map is a map image with pixel-by-pixel subtraction of an early-phase image from a delayed maximum enhancement phase image obtained in a dynamic study. The sensitivity and specificity were calculated with or without subtraction in the diagnosis of benign and malignant breast tumors.
RESULTS:
The sensitivity of benign breast masses increased from 0.879 to 0.939, and the specificity increased from 0.818 to 0.909 with reference to the DPS map. There was statistical difference between with or without DPS (Z=2.023, P=0.043). The sensitivity of breast malignant masses increased from 0.889 to 0.933, and the specificity increased from 0.867 to 0.911 with reference to the DPS map, with statistical difference between with or without DPS map (Z=2.294, P=0.021). The pattern of TIC changed from continuous to a plateau in 8 patients, from a plateau to washout in 10, and from continuous to washout in 5. No changes were observed in the other 55 patients.
CONCLUSION
MR Subtraction is a simple and useful technique to identify breast lesions. It helps to accurately set the location of the ROI TIC and improve the detection rate of benign and malignant breast tumors.
Breast Neoplasms
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diagnosis
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Contrast Media
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Diagnosis, Differential
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Female
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Humans
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Image Enhancement
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Magnetic Resonance Imaging
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Retrospective Studies
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Sensitivity and Specificity
8.MR perfusion imaging of the liver: early findings after transcatheter arterial chemoembolization of hepatocellular carcinoma
Dujun BIAN ; Enhua XIAO ; Yunping XIAO ; Xiangyu CHEN ; Weijun SITU ; Zhong HE ; Shuwen YUAN ; Jianning SUN
Chinese Journal of Radiology 2010;44(12):1248-1252
Objective To investigate the value of MR perfusion imaging in early detection of findings following arterial chemoembolization of hepatocellular carcinoma Methods Twenty eight consecutive patients with pathologically-confirmed HCC were evaluated. All patients underwent MR perfusion imaging at pre-TACE and 3 to 10 days after TACE. The negative enhancement integral (NEI) ,the time to peak(TTP) ,the maximum slope of decrease (MSD) , the signal enhance ratio (SER) were acquired from MRI software FuncTool 2. 5.36a Version. Statistical analysis using SPSS 14, least significant difference test (t test) were utilized. Results The time intensive curve of tumor was observed to descend rapidly to reach the peak at pre-TACE studies, whereas it descended slowly to reach the peak on post TACE studies. The Value of TTP and SER prior to TACE were(51.2 ± 10. 3) s, 60. 6 ± 36. 3 respectively, and post TACE (43.7 ± 12. 0)s, 41.2 ±27. 5 respectively. The values of TTP and SER post TACE were lower than those prior to TACE (P < 0. 05). The value of NEI prior to TACE was 108.7 ± 58.9, and after TACE 149. 6 ±80. 1 and there was statistically significant difference (P <0. 05). The Value of MSD post TACE were lower than those prior to TACE, but there was no statistical significance (P > 0. 05). Conclusion PWI is a very sensitive imaging technique that can be used to monitor early dynamic changes of HCC following TACE.
9.Magnetic resonance perfusion imaging before and after transcatheter arterial chemoembolization treatment of hepatocellular carcinoma
Duun BIAN ; Enhua XIAO ; Dongxu HU ; Yunping XIAO ; Xiangyu CHEN ; Weijun SITU ; Zhong HE ; Shuwen YUAN ; Jianning SUN
Chinese Journal of Medical Imaging Technology 2010;26(1):89-92
Objective To observe the change of MR perfusion value in patients with hepatocellular carcinoma (HCC) before and after transcatheter arterial chemoembolization (TACE). Methods A total of 22 patients with HCC underwent MR perfusion weighted imaging (MR PWI) before TACE and 3-10 days after TACE. The mean time to enhance (MTE), negative enhancement integral (NEI), time to peak (TTP) and maximum slope of decrease (MSD) before and after TACE were acquired and compared. Results The time intension curve (TIC) of HCC region was observed to descend rapidly before TACE, while descended slowly after TACE. The value of MTE and TTP after TACE were lower than those before TACE (P<0.05), and the value of NEI after TACE was higher than that before TACE (P<0.05). The value of MSD after TACE were lower than that before TACE, but no statistical significance was found (P>0.05). Conclusion MR PWI is a very sensitive imaging technique that be used to monitor blood flow changes of HCC before and after TACE and evaluate efficacy of TACE.
10.Ciprofloxacin Combined with Amikacin to Reduce Drug-resistant Mutants of Escherichia coli
Mingtao LIU ; Enhua SUN ; Shaojie BI ; Yu LI
Chinese Journal of Nosocomiology 2009;0(17):-
OBJECTIVE To study whether the ciprofloxacin(CIP)combined with amikacin(AMK)will decrease the drug-resistant mutants of Escherichia coli(ECO)in vitro.METHODS The MIC of CIP and AMK alone was determined by agar plates dilution method,and the combined MIC by checkerboard method.The mutant prevention concentration(MPC)both alone and combined was determined by the method of agar plates dilution method,and then the value of selectiveity index(SI)(MPC/MIC)would be acquired.RESULTS The SI of two antibiotics separatedly were 16(CIP)and 32(AMK).When two antibiotics combined,if the concentration of AMK and CIP reached 2MIC could restrain the mutants.After these two antibiotics combined,the value of SI(MPCcombined/MICcombined)came to be 8(0.008/0.001)and 8(2.0/0.25).CONCLUSIONS Compared with alone,CIP and AMK combined can decrease the MPC and SI to ECO,and have the more effect to AMK.In this way,we can reduce the drug-resistant mutants.

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